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Sample records for early staphylococcal scalded

  1. Staphylococcal scalded skin syndrome hos voksne

    DEFF Research Database (Denmark)

    Mikkelsen, Carsten Sauer; Mikkelsen, Dorthe Bisgaard; Jensen, Thøger Gorm

    2010-01-01

    We describe a case of adult staphylococcal scalded skin syndrome (SSSS) in a 91-year-old woman. The patient had a Staphylococcus aureus infection in her right ear with redness and yellow secretion. A few days later she developed Nikolsky sign-positive tender enlarged flaccid bullae involving most...

  2. Congenital staphylococcal scalded skin syndrome in a premature infant

    NARCIS (Netherlands)

    Haveman, LM; Fleer, A; de Vries, LS; Gerards, LJ

    2004-01-01

    A case of congenital staphylococcal scalded skin syndrome (SSSS) with fatal outcome in a premature infant is reported. An intrauterine infection with Staphylococcus aureus was probably the cause for the fulminant course of the disease. Despite adequate antibiotic treatment, the infant died within 24

  3. Neonatal staphylococcal scalded skin syndrome: clinical and outbreak containment review.

    LENUS (Irish Health Repository)

    Neylon, Orla

    2012-01-31

    Staphylococcal scalded skin syndrome (SSSS) is a toxin-mediated exfoliating skin condition predominated by desquamation and blistering. Neonatal outbreaks have already been reported; however, our outbreak highlights the potential for SSSS following neonatal health promotion measures such as intra-muscular vitamin K administration and metabolic screening (heel prick) as well as effective case containment measures and the value of staff screening. Between February and June 2007, five confirmed cases of neonatal SSSS were identified in full-term neonates born in an Irish regional maternity hospital. All infants were treated successfully. Analysis of contact and environmental screening was undertaken, including family members and healthcare workers. Molecular typing on isolates was carried out. An outbreak control team (OCT) was assembled and took successful prospective steps to prevent further cases. All five Staphylococcus aureus isolates tested positive for exfoliative toxin A, of which two distinct strains were identified on pulsed-field gel electrophoresis analysis. Two cases followed staphylococcal inoculation during preventive measures such as intra-muscular vitamin K administration and metabolic screening (heel prick). None of the neonatal isolates were methicillin resistant. Of 259 hospital staff (70% of staff) screened, 30% were colonised with S. aureus, and 6% were positive for MRSA carriage. This is the first reported outbreak of neonatal SSSS in Ireland. Effective case containment measures and clinical value of OCT is demonstrated. Results of staff screening underlines the need for vigilance and compliance in hand disinfection strategies in maternity hospitals especially during neonatal screening and preventive procedures.

  4. Regional outbreak of staphylococcal scalded skin syndrome in healthy children

    Directory of Open Access Journals (Sweden)

    Hyun Jeong Do

    2010-01-01

    Full Text Available Purpose : Staphylococcal scalded skin syndrome (SSSS is a relatively uncommon superficial blistering skin disease that is due to Staphylococcus aureus. We had experienced a regional outbreak of SSSS over 3 years in healthy children. Methods : We retrospectively reviewed the medical records of those patients diagnosed as SSSS. Most of neonatal cases were nosocomial infections and excluded from the analysis. The clinical features, laboratory findings, the isolation and antibiotic resistance of S. aureus, the antibiotic management and other supportive treatments were analyzed. Results : Fifty-five patients with SSSS were admitted to our hospital from October 2001 to September 2004. The median age of patients was 3.0 years. Of the 55 patients, 9 were the generalized type, 13 were the intermediate type and 33 were the scarletiniform rash. All the patients were living in neighborhood of the Jinju area. S. aureus were isolated from 9 of the patients and all of the isolated S. aureus were methicillin resistant. All the patients except two were treated with intravenous flocloxacillin or nafcillin and/or cefotaxime. All the patients recovered during the follow-up period of 2 to 3 weeks. Conclusion : We experienced a regional outbreak of SSSS in previous healthy children. Further study for finding the carriers of S. aureus caused SSSS and preventing the spread of this disease is needed. Additionally, guidelines for treating SSSS due to methicillin resistant S. aureus should be established.

  5. STAPHYLOCOCCAL SCALDED SKIN SYNDROME:RETROSPECTIVE ANALYSIS OF 82 CASES

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective To explore distinctive clinical manifestations and appropriate treatment, and assess prognosis of staphylococcal scalded skin syndrome ( SSSS). Methods A retrospective analysis was conducted of the data of 82 cases of SSSS hospitalized at Xinhua Hospital during the period from May 1993 to September 2003.Results The disease in all the 82 patients occurred in their first decade ( mean 2.5 years). Possible predisposing factors were found in 48 (58. 5% ). Fever was present in 78 (95. 1% ). Radial spokes of crusting around mouth were present in 80 (97. 6% ). Erythema began on the face, especially around the mouth and eye in 63 (76. 8% ).The course was acute in all cases and the eruptions quickly spread to the whole body within one day to two days. Of the 82 cases of SSSS, 47 were complete form of SSSS, 27 were abortive form of SSSS, and 8 were between the two forms. Staphylococcus aureus with positive staphylocoagulase was isolated from the possible primary infection sites including pharynx, eyelid, conjunctiva, nose, ear, and skin in 18 of 31 patients. Microbiological cultures of bullae and little pustulae developed after the onset were negative in 16 cases. All the 82 patients completely recovered after receiving antibiotic therapy ( ceftriaxone, oxacillin ) alone or in combination with human immunoglobulin (IVIG) therapy. Additional IVIG therapy was used in those patients who had systemic involvements such as pneumonia, fever higher than 38. 5℃ or leukocytosis. Conclusion SSSS is a spectrum disease. Besides abortive and complete forms, presenting between the two forms a new form might be appeared in 8 cases who developed both scarlatiniform rash and flaccid bullae. The abortive form and complete form are usually misdiagnosed clinically. Radial spokes of crusting around mouth seem to be characteristic manifestation of SSSS. All the patients in this study had favorable prognosis after receiving prompt diagnosis and appropriate treatment.

  6. Staphylococcal Scalded Skin Syndrome in Child. A Case Report and a Review from Literature

    Directory of Open Access Journals (Sweden)

    Grama Alina

    2016-10-01

    Full Text Available Staphylococcal scalded skin syndrome (SSSS is the medical term used to define a skin condition induced by the exfoliative toxins produced by Staphylococcus aureus. The disorder is also known as Ritter disease, bullous impetigo, neonatal pemphigus, or staphylococcal scarlet fever. The disease especially affects infants and small children, but has also been described in adults. Prompt therapy with proper antibiotics and supportive treatment has led to a decrease in the mortality rate.

  7. Staphylococcal scalded skin syndrome : loss of desmoglein 1 in patient skin

    NARCIS (Netherlands)

    Aalfs, A. Susanne; Oktarina, D. A. Mira; Diercks, Gilles F. H.; Jonkman, Marcel F.; Pas, Hendri H.

    2010-01-01

    Staphylococcal scalded skin syndrome (SSSS) is a blistering disease of the skin caused by an infection with certain strains of Staphylococcus aureus. In vitro studies have suggested that exfoliative toxins secreted by these bacteria cleave the desmosomal adhesion molecule desmoglein 1 leading to los

  8. Staphylococcal Scalded-Skin Syndrome Complicating Wound Infection in a Preterm Infant with Postoperative Chylothorax

    Science.gov (United States)

    Peters, Bjoern; Hentschel, Juliane; Mau, Harald; Halle, Elke; Witte, Wolfgang; Obladen, Michael

    1998-01-01

    The course of infection in a 3-week-old premature newborn suffering from extensive dermatitis with flaccid blisters is described. Staphylococcus aureus was recovered from a local wound infection around a chest tube inserted to drain a postoperative chylothorax. The strain isolated tested positive for the eta gene for exfoliative toxin A, the causative agent of staphylococcal scalded-skin syndrome (SSSS). In this case, prematurity and loss of chylus with consecutive lymphopenia may have contributed to development of SSSS. PMID:9738069

  9. Staphylococcal scalded skin syndrome in a premature newborn caused by methicillin-resistant Staphylococcus aureus: case report

    Directory of Open Access Journals (Sweden)

    Andreas Hörner

    Full Text Available CONTEXT: Staphylococcal scalded skin syndrome is an exfoliative skin disease. Reports of this syndrome in newborns caused by methicillin-resistant Staphylococcus aureus are rare but, when present, rapid diagnosis and treatment is required in order to decrease morbidity and mortality. CASE REPORT: A premature newly born girl weighing 1,520 g, born with a gestational age of 29 weeks and 4 days, developed staphylococcal scalded skin syndrome on the fifth day of life. Cultures on blood samples collected on the first and fourth days were negative, but Pseudomonas aeruginosa and Enterococcus sp. (vancomycin-sensitive developed in blood cultures performed on the day of death (seventh day, and Pseudomonas aeruginosa and Serratia marcescens were identified in cultures on nasopharyngeal, buttock and abdominal secretions. In addition to these two Gram-negative bacilli, methicillin-resistant Staphylococcus aureus was isolated in a culture on the umbilical stump (seventh day. The diagnosis of staphylococcal scalded skin syndrome was based on clinical criteria.

  10. STAPHYLOCOCCAL SCALDED SKIN SYNDROME MIMICKING TOXIC EPIDERMAL NECROLYSIS IN A HEALTHY ADULT

    Directory of Open Access Journals (Sweden)

    Tomoko Oishi

    2013-07-01

    Full Text Available Introduction: Staphylococcal scaled skin syndrome (SSSS presents generalized form bullous impetigo caused by Staphylococcus aureus (S. aureus infection, typically seen in infants and children. SSSS may occur also in adults; however, the majority of adult cases are those with immunosuppression. Atypical clinical features of impetigo in adults sometimes make it difficult to diagnose correctly. Case Report: A 74-year-old healthy woman was hospitalized, complaining of extensive desquamative erythema and a number of erosions. She was administered oral antiviral drugs under suspicion of herpes zoster prior to 10 days. Initial diagnosis on the admission was toxic epidermal necrolysis (TEN due to antiviral tablets; however, steroid pulse therapy resulted in no effect. Bacterial culture yielded coagulase-positive methicillin-resistent S. aureus, producing exfoliative toxin B. A biopsy specimen showed subcorneal splitting of the epidermis. The diffuse erosions gradually improved over 10 days by the treatment with intravenous antibiotics. Conclusions: The differentiation between streptococcal scaled skin syndrome (SSSS and TEN is sometimes difficult. It is important to remind SSSS when we suspect TEN, even in healthy adults..

  11. Early childhood severe scalds in a developing country: A 3-year retrospective study

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    Pius Agbenorku

    2013-12-01

    Full Text Available The burns intensive care unit (BICU staff observed an increasing number of pediatric scald burn admissions as a result of increase injuries associated with the scald burns. A retrospective study was conducted to identify scalds demographics, etiologies, and mortality risk factors. This descriptive study comprised a total of 166 patients aged 0-5 years, who were admitted to the BICU of the Reconstructive Plastic Surgery and Burns Unit (RPSBU through the Accident and Emergency (A and E Centre of the Komfo Anokye Teaching Hospital (KATH from May 1 st 2009 to April 30 th 2012. Source of information was the BICU Computerized Database System. Data extracted included demographics as well as treatment methods and outcomes. The study population was 166; 92 (55.4% males and 74 (44.6% females. Scalds admissions were 141 (84.9%; 13 (9.2% of them died, 83 (58.9% discharged, and 45 (31.9% transferred-out to another burn ward and pediatric surgery ward in the hospital. Scald patients' demographics included 78 males (55.3% and 63 females (44.7%; mean age was 2.18 years. Mortality risk factors identified were age <3 years (P = 0.044; scalds from hot water (P = 0.033, total burns surface area >30% (P = 0.017, and multiple body parts affected (P = 0.049. The current study showed age, hot water, and Total Burns Surface Area (TBSA as risk factors of early childhood scalds. Education on scalds prevention targeting mothers/caregivers is needed to create awareness of the frequency, severity, and danger associated with pediatric scalds.

  12. Antimicrobial agent of susceptibilities and antiseptic resistance gene distribution among methicillin-resistant Staphylococcus aureus isolates from patients with impetigo and staphylococcal scalded skin syndrome.

    Science.gov (United States)

    Noguchi, Norihisa; Nakaminami, Hidemasa; Nishijima, Setsuko; Kurokawa, Ichiro; So, Hiromu; Sasatsu, Masanori

    2006-06-01

    The susceptibilities to antimicrobial agents of and distributions of antiseptic resistance genes in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated between 1999 and 2004 in Japan were examined. The data of MRSA strains that are causative agents of impetigo and staphylococcal scalded skin syndrome (SSSS) were compared with those of MRSA strains isolated from patients with other diseases. The susceptibilities to antiseptic agents in MRSA isolates from patients with impetigo and SSSS were higher than those in MRSA isolates from patients with other diseases. The distribution of the qacA/B genes in MRSA strains isolated from patients with impetigo and SSSS (1.3%, 1/76) was remarkably lower than that in MRSA strains isolated from patients with other diseases (45.9%, 95/207). Epidemiologic typings of staphylococcal cassette chromosome mec (SCCmec) and pulsed-field gel electrophoresis (PFGE) showed that MRSA strains isolated from patients with impetigo and SSSS had type IV SCCmec (75/76), except for one strain, and 64.5% (49/76) of the strains had different PFGE types. In addition, the patterns of restriction digestion of all tested qacA/B plasmid in MRSA isolates having different PFGE types were identical. The results showed that a specific MRSA clone carrying qacA/B was not prevalent, but qacA/B was spread among health care-associated MRSA strains. Therefore, it was concluded that the lower distribution rate of qacA/B resulted in higher susceptibilities to cationic antiseptic agents in MRSA isolated from patients with impetigo and SSSS.

  13. Scald Burns

    Science.gov (United States)

    Safety Tips & Info Scald Burns Thousands of scald burns occur annually, and ALL are preventable! The two high-risk populations are children under the age ... the single most important factor in preventing scald burns. Increased awareness is the key to scald prevention! ...

  14. Effective Treatment of Staphylococcal Scalded Skin Syndrome with Platelet Microbicidal Protein in CBRB-Rb(8.17)1Iem Mice Model.

    Science.gov (United States)

    Miroshnikov, Sergey A; Gritsenko, Viktor A; Ivanov, Iuri B

    2015-09-01

    Skin and soft-tissue infections are among the most common infections. Staphylococcus aureus may cause a number of toxin-mediated diseases, including staphylococcal scalded skin syndrome (SSSS). The therapeutic efficacy of some antimicrobial peptides was recently evaluated in a mouse model of SSSS. This study is the first in vivo demonstration of the use of PMP to improve outcome of SSSS. Twenty-four CBRB-Rb(8.17)1Iem female mice naturally infected by endogenous S. aureus with SSSS symptoms were used in this work and divided into two equal groups. From neck of each mouse was isolated and identified endogenous exfoliative producing strain of S. aureus. PMP was obtained from human platelets and tested against Bacillus subtilis ATCC 6633. PMP had bactericidal activity against B. subtilis ATTC 6633 and endogenous strain of S. aureus at 2.0 ± 0.5 and 14.5 ± 0.5 µg/ml, respectively. At 4 weeks, the mice of experimental group were treated subcutaneous near exfoliative zone with 0.2 ml of PMP in final concentration 10 µg/ml every day. Control mice was injected with 0.2 ml 0.9% NaCl. At 1 day of experiment maximal zone of alopecia was at PMP-treating group (380 ± 20 mm(2)) in comparison with control group (167 ± 10 mm(2), p < 0.01). At 50 day of observation (22nd day after the end of treatment), the square of alopecia in control group was 1220 ± 40 mm(2) in comparison with 870 ± 17 mm(2) in experimental group (p < 0.01). The antistaphylococcal in vivo activity of PMP demonstrated in present study makes these molecules potentially useful for treatment of SSSS.

  15. CLINICAL TREATMENT METHODS OF 42 CASES OF STAPHYLOCOCCAL SCALDED SKIN SYNDROME AND THEIR EFFECTS%葡萄球菌性烫伤样皮肤综合征42例的临床治疗方法及其效果分析

    Institute of Scientific and Technical Information of China (English)

    黄英举; 郑小帆; 蓝莉; 刘卉

    2011-01-01

    [Objective] To summarize the clinical symptoms of staphylococcal scalded skin syndrome, and to explore its treatment. [Methods] 42 children of staphylococcal scalded skin syndrome in our hospital were selected, summarized the clinical performance and characteristics, and analyzed the two different treatment methods. [Results] The staphylococcal scalded skin syndrome, once diagnosed, must be given adequate and timely antibiotic treatment; 27 patients in the control group were given the conventional treatment, besides, the use of adrenal corticosteroids dexamethasone therapy combined with support treatment. Children with fever clearance time, erythema shrinking time, epidermal stripping bare area drying time, tenderness fade time and cure time were significantly superior to the control group, the difference was significant (P< 0.05). [Conclusion] The staphylococcal scalded skin syndrome is a serious infectious disease. The adequate antibiotic treatment according to symptoms, at the same time, the supporting treatment combined with adrenal cortex hormone therapy and strengthening the care on children's mouths and eyes, have an important role in their early recovery.%[目的]总结葡萄球菌烫伤样皮肤综合征的临床症状并探讨其治疗方法.[方法]选取多年来某院收治的葡萄球菌性烫伤样皮肤综合征患儿病例42例,总结其临床表现及特征,并对两种不同的治疗方法的疗效进行分析.[结果]葡萄球菌烫伤样皮肤综合征一经确诊,需及时给予足量的抗生素治疗;同时本研究观察组27例在对照组常规处理的基础上.采用肾上腺皮质激素类药物地塞米松联合支持疗法进行治疗.患儿退热时间、红斑萎缩时间、表皮剥裸区干燥时间、触痛消退时间及痊愈出院治疗时间均明显优干常规对照组,差异具有统计学意义(P<0.05).[结论]葡萄球菌性烫伤样皮肤综合征是一种严重的感染性疾病,对症足量的抗生素治疗同

  16. Effect of nonpeptide NK1 receptor antagonist L-703,606 on the edema formation in rats at early stage after deep partial-thickness skin scalding

    Institute of Scientific and Technical Information of China (English)

    Ke Tao; Hong-Tao Wang; Bi Chen; Bo-Tao Wang; Zhi-Yuan Li; Xiong-Xiang Zhu; Chao-Wu Tang; Da-Hai Hu

    2013-01-01

    Objective: To investigate the effect and the relevant potential mechanism of nonpeptide neurokinin 1 (NK1) receptor antagonist L-703,606 in the edema formation after burn injury.Method:L-703,606 treatment was performed in Sprague-Dawley (SD) rats at early stage after deep partial-thickness skin scalding. One hundred and fifty two adult male SD rats were used in the study and randomly divided into sham scald (SS, n=8), scald control (SC, n=48), and L-703,606 treatment (LT, n=48) groups. The rats in SC and LT groups were subjected to 20% total body surface area (TBSA) deep partial-thickness skin scalding. Modified Evans blue extravasation, tracing electron microscopy by lanthanum nitrate and mean water content assay were employed to observe and detect the changes of vascular permeability, ultrastructure and edema formation in adjacent tissue to the wounds and in the jejuna of rats at early stage (72 h) after scald. Results:The pathological increase of vascular permeability in the periwound tissue and jejunum of rats in LT group were significantly lower than that in SC group (P<0.01), and recuperated earlier. Meanwhile, the changes of water contents of corresponding tissues in LT group were lighter than those in SC group (P<0.01). The ultrastructural changes of the microvessels in the peri-wound tissue of LT group showed that the junctions between microvascular endothelium cells were more narrow than those of SC group, moreover, and the number of opening and the engorgement and cavitation of the vascular endothelium cells decreased, the areosis and edema in perivascular tissue lightened, and the precipitation of the high eletron density lanthanum tracing agent in the interspace of the tissue decreased significantly in LT group. Conclusions: It is concluded that nonpeptide NK1-receptor antagonist L-703,606 could lighten the vascular permeability and edema formation in the periwound tissue and jejunum, and accelerate the normalization process of pathological changes in

  17. 葡萄球菌性烫伤样皮肤综合征表皮剥脱毒素分型及耐药分析%The Exfoliative Toxin Serotypes and Resistance of Staphylococcal Aureus from Staphylococcal Scalded Skin Syndrome

    Institute of Scientific and Technical Information of China (English)

    赵俊英; 王晓彦

    2014-01-01

    目的 探讨引起本地区46例葡萄球菌性烫伤样皮肤综合征(staphylococcal scalded skin syndrome,SSSS)患者的金黄色葡萄球菌(staphylococcus aureus,SA)所分泌的表皮剥脱毒素(exfoliative toxin,ET)血清型及SA的耐药情况,并与寻常型脓疱疮及皮肤疖肿组进行对比.方法 138株SA分离自SSSS(46株),寻常型脓疱疮(49株),皮肤疖肿组(43株).应用多重PCR方法检测SA产生的ET类型,应用Kirby-Bauer纸片法检测SA对15种抗生素的耐药情况.结果 SSSS组SA共46株(100.00%)均产生ET,均是以单产ETB为主;寻常型脓疱疮组产生ETB的有30株(61.22%);皮肤疖肿组无一株产生ETB.138株SA均未检测到ETA或ETD的表达.三组SA分泌ET的比例差异存在统计学意义(P<0.05).SSSS、寻常型脓疱疮及皮肤疖肿组对青霉素、红霉素、克林霉素的耐药率较高,而对一、二代头孢类抗生素较敏感.三组共发现6株耐甲氧西林金黄色葡萄球菌(M RSA).结论 内蒙古呼和浩特市地区SSSS的致病SA主要以单产ETB为主.本地区SA主要对青霉素、红霉素、克林霉素高度耐药,对一、二代头孢类抗生素较敏感.

  18. Scalded skin syndrome

    Science.gov (United States)

    ... Severe bloodstream infection ( septicemia ) Spread to deeper skin infection ( cellulitis ) When to Contact a Medical Professional Call your provider or go to the emergency room if you have symptoms of this disorder. Prevention ... Alternative Names Ritter disease; Staphylococcal ...

  19. 四肢浅Ⅱ度烫伤患者早期创面冷疗临床效果对比观察%Limb superifcial II degree scald wound therapy in patients with early cold treatment clinical effect observation

    Institute of Scientific and Technical Information of China (English)

    黄新红; 李涛; 刘智文

    2014-01-01

    目的:探讨四肢浅Ⅱ度烫伤患者创面早期处理方法。方法:选择40例四肢浅Ⅱ度烫伤患者,其中20例患者创面早期使用持续冷疗,另20例患者烫伤创面进行传统处理:冷水冲洗后涂抹烧伤膏暴露疗法,对比治疗后疼痛、炎性反应水平、创面渗出停止时间、治疗费用、住院时间、患者满意度等。结果:四肢浅Ⅱ度烫伤患者早期创面冷疗临床治疗效果更为确切肯定。结论:冷疗的作用机制及如何进一步规范治疗的细节。%Objective To study the limb superficial II degree burn scald patients early treatment method.Methods Choose 40 cases of superficial second degree scald patients limbs,in which 20 cases of early wound using continuous cold therapy,another 20 patients with burn wound of traditional processing:cold water rinse after smearing burn ointment exposure therapy,treatment of pain after contrast,inflammatory reaction, wound exudate level stop time,treatment costs,time of hospitalization,patients satisfaction.Results The limb superficial II degree scald wound therapy in patients with early cold treatment clinical effect more sure exactly.Conclusion The mechanism of cold treatment and how to further regulate the details of treatment.

  20. Scalded skin syndrome: Diagnosis, differential diagnosis, and management of 42 children.

    Science.gov (United States)

    Margileth, A M

    1975-04-01

    The bacteriologic, epidemiologic, clinical, and diagnostic findings and management of the scalded skin syndrome (SSS) in 42 children are reported SSS may present in one of three ways: (1) Ritter's disease in infants, characterized by an acute, generalized bullous dermatitis simulating the appearance of scalded skin, followed by exfoliation; (2) Lyell's disease, or toxic epidermal necrolysis (TEN), in children or adults, with scalded skin and bullae followed by extensive exfoliation; or (3) a nonstreptococcal scarlatiniform eruption, or staphylococcal scarlet fever, manifested by a generalized scarlatiniform erythema ans subsequent minimal to moderate fine desquamation. Staphylococcus aureus was isolated and implicated as the causative agent in 32 of the 42 children; nine of the other ten children had received a drug or vaccine preceding their rash. SSS in children has increased in frequency recently, is usually due to S aureus infection with potential epidemic aspects, can be recognized easily by its unusual clinical features, and has an excellent prognosis with appropriate management.

  1. Staphylococcal scalded skin syndrome due to burn wound infection.

    Science.gov (United States)

    Farroha, A; Frew, Q; Jabir, S; Dziewulski, P

    2012-09-30

    Introduction. Le syndrome de la peau ébouillantée staphylococcique (sigle anglais conventionnel, SSSS) est une exfoliation aiguë de la peau causée par des toxines A et B. Bien que le Staphylococcus aureus soit une cause fréquente d’infection des brûlures, la SSSS suite à une infection brûlure est rare. Méthode. Les Auteurs ont effectué une revue rétrospective de tous les cas de patients atteints de SSSS hospitalises admis dans un service régional des brûlures entre janvier 2008 et janvier 2012. Résultats. Deux cas de SSSS ont été signalés au cours de cette période qui se sont produits suite à une brûlure. Le premier cas était un garçon de 17 mois qui avait été hospitalisé pour un traitement conservateur pour ébouillantement dans 6% de la surface corporelle totale de profondeur variable. Le quatrième jour, il a développé une exfoliation dans 85% de la surface corporelle. Quant au deuxième cas, il s’agissait d’un garçon de dix mois qui a subi une brûlure de 1% de la surface corporelle et qui a été traité en manière conservatrice dans la communauté par son médecin généraliste. Le cinquième jour, il a développé une exfoliation dans 80% de la surface corporelle. Le Staphylococcus aureus a été isolé qui provenait des brûlures dans les deux cas. Conclusion. Ces deux cas montrent qu’il est essentiel que les brûlologues et les spécialistes des soins intensifs soient au courant de la possibilité de la présence de SSSS chez des patients souffrant de brûlures, avec tous ses potentiels effets dévastateurs.

  2. Exfoliative toxin serotype genes and antibiotic resistance of staphylococcus aureus isolated from children with staphylococcal scalded skin syndrome%葡萄球菌性烫伤样皮肤综合征的致病金黄色葡萄球菌剥脱毒素基因型及耐药研究

    Institute of Scientific and Technical Information of China (English)

    苏怡帆; 王华; 肖异珠; 欧阳莹; 李咏梅; 罗晓燕; 谭琦; 向娟

    2008-01-01

    Objective To investigate the exfoliative toxin serotype genes and antibiotic resistance of Staphylococcus aureus (SA) isolated from children with staphylococcal scalded skin syndrome (SSSS). Methods In total, 108 strains of SA were isolated from 36 patients with SSSS, 36 patients with impetigo and 36 patients with abscess. Multiplex PCR was used to detect the staphylococcal exfoliative toxin A, B and D genes, Kirby-Baner method to test the susceptibilities of SA strains to 20 antibiotics. Results All the 36 SA isolates from SSSS patients were ET-positive, and 2 (6%) produced ETA, 7 (19%) ETB, 27 (75%) both ETA and ETB; of the 36 isolates from patients with impetigo, 78% produced ET, and 14% produced ETA, 64% produced both ETA and ETB, while no single ETB-producing strain was found; ET was detected in only one (2.8%) SA isolate from abscess patients, which produced both ETA and ETB. ETD was detected in none of the SA isolates. There was a statistical difference in the distribution of ET serotype among the three diseases (χ2=89.4, P < 0.01) and the proportion of ET-producing strains in SSSS group was signifi-cantly higher than that in impetigo group (χ2=9.0, P < 0.01) and abscess group (χ2= 68.1, P < 0.01). All the SA isolates were highly resistant to penicilin, ampicillin, macrolides and clindamycin, but sensitive to other 15 common antibiotics such as cephalosporin. Two strains of MRSA were found in patients with abscess. Conclusion In Chongqing, ET-producing SA is the common pathogenic bacteria of SSSS and impetigo, and most of SA strains produce both ETA and ETB.%目的 探讨葡萄球菌性烫伤样皮肤综合征(SSSS)致病的金黄色葡萄球菌(简称金葡菌)分泌表皮剥脱毒素(ET)的主要类型及耐药情况.方法 108株金葡菌分离自SSSS(36例)、脓疱疮(36例)及皮肤脓肿(36例).用多重PCR法测定致病金葡菌菌株产ETA、ETB、ETD基因型,Kirby-Bauer纸片法检测其对20种抗生素的敏感性.结果 36

  3. Metabolomic change precedes apple superficial scald symptoms.

    Science.gov (United States)

    Rudell, David R; Mattheis, James P; Hertog, Maarten L A T M

    2009-09-23

    Untargeted metabolic profiling was employed to characterize metabolomic changes associated with 'Granny Smith' apple superficial scald development following 1-MCP or DPA treatment. Partial least-squares discriminant analyses were used to link metabolites with scald, postharvest treatments, and storage duration. Models revealed metabolomic differentiation between untreated controls and fruit treated with DPA or 1-MCP within 1 week following storage initiation. Metabolic divergence between controls and DPA-treated fruit after 4 weeks of storage preceded scald symptom development by 2 months. alpha-Farnesene oxidation products with known associations to scald, including conjugated trienols, 6-methyl-5-hepten-2-one, and 6-methyl-5-hepten-2-ol, were associated with presymptomatic as well as scalded control fruit. Likewise, a large group of putative triterpenoids with mass spectral features similar to those of ursolic acid and beta-sitosterol were associated with control fruit and scald. Results demonstrate that extensive metabolomic changes associated with scald precede actual symptom development.

  4. 小儿烧烫伤早期并发腹泻的治疗分析%The Treatment Analysis of Burn and Scald Children at Early Stage and Complicated with Diarrhea

    Institute of Scientific and Technical Information of China (English)

    考玉芹

    2016-01-01

    目的:探讨小儿烧烫伤并发腹泻病因、早期治疗方法,提高救治水平。方法:对2008~2014年我院共收治32例大面积烧伤早期并发腹泻患儿,根据其早期临床表现、病因、治疗经过及预后进行分析总结,采用烧伤早期抗休克补液及腹泻补液、检测调节电解质、抗感染、维持肠道正常菌群、营养支持、创面换药等治疗措施。结果:在烧烫伤早期快速液体复苏过程中患儿出现腹泻,根据患儿腹泻原因同时给予抗休克、腹泻补液及止泻、调整胃肠道微环境,创面换药,患儿预后效果良好。结论:烧烫伤患儿早期并发腹泻,经过积极抗休克、合理补液、抗感染、创面包扎换药、湿润疗法、适当使用肠道微生态制剂、保护胃肠黏膜制剂后,患儿平稳度过休克期,增加成活率。%Objective:To investigate the pathogeny and early treatment of burn and scald children complicated with diarrhea, in order to improve the level of treatment. Methods:From 2008 to 2014, our hospital had received 32 cases of children with extensive burned at early stage and complicated with diarrhea, according to the analysis and summary of their early clinical manifestations, pathogeny, treatment procedure and prognosis, treated by anti-shock and diarrhea fluid infusion, testing and regulating electrolytes, anti-infection, maintaining normal intestinal flora, nutritional support, wound dressing. Results:The children had appeared diarrhea in the process of rapid fluid resuscitation at the early stage of burn and scald, according to the diarrhea reasons and treated by anti-shock and diarrhea fluid infusion, anti-diarrhea, regulating micro environment of gastrointestinal tract and wound dressing, the children's prognosis were good. Conclusion:For the burn and scald children at early state and complicated diarrhea, after the treatment of anti-shock, reasonable fluid infusion, anti-infection, wound dressing

  5. Protective effect of early application of lytic cocktail on small intestine of severely scalded rats%早期应用冬眠合剂对严重烫伤大鼠小肠的保护作用

    Institute of Scientific and Technical Information of China (English)

    邵庆波; 章雄; 陈雪莲; 刘琰; 张勤; 廖镇江

    2010-01-01

    .01);烫伤+冬眠合剂组各时相点IL-10水平均高于烫伤组,并在6、24 h时差异具有统计学意义(F值分别为8.668、19.634,P<0.05或P<0.01).结论 早期应用冬眠合剂可减轻严重烫伤大鼠小肠黏膜水肿和损害,该机制可能与其降低肠道ICAM-1的表达和血液炎症介质水平、减少肠道局部炎症细胞数量有关.%Objective To study the protective effect of early application of lytic cocktail on small intestine of severely scalded rats. Methods Sixty-six male SD rats were divided into sham injury group (SI, n =6) , scald group (S, n = 30) and scald + lytic cocktail group (SL, n =30) according to the random number table. After anesthesia, rats in the latter 2 groups were inflicted with 30% full-thickness scald, while rats in S group were sham scalded with 37 ℃ water. Resuscitation was carried out by intraperitoneal injection with 2 mL · kg-1 · %TBSA-1 lactated Ringer's solution in all rats; meanwhile 12 mL/kg lytic cocktail [ 1 mL pethidine (50 mg/mL) + 1 mL chlorpromazine (25 mg/mL) + 1 mL promethazine (25 mg/mL) + 125 mL saline] was hypodermically injected to rats in SL group, while 12 mL/kg saline was injected into rats in the other 2 groups. Samples of blood and small intestine were harvested from S and SL groups at post scald hour (PSH) 3, 6, 12, 24, 48 and from SI group at PSH 3, with 6 rats in each group at each time point. Pathological changes in intestine were observed, and the expression of intercellular adhesion molecule 1 (ICAM-1) and CD68 were determined with immunohistochemistry at PSH 24 for S and SL groups and at PSH 3 for SI group. Plasma levels of D-lactate, diamine oxidase (DAO) , IL-1β, TNF-α, IL-10 were determined with ELISA. Data were processed with one-way analysis of variance. Results (1) At PSH 24, mild hemorrhage, inflammatory cell infiltration and epithelial cell shedding were observed in small intestinal mucosa of rats in S group.Compared with S group, the intestinal villi of SL group

  6. Early staphylococcal biofilm formation on solid orthopaedic implant materials: in vitro study.

    Directory of Open Access Journals (Sweden)

    Hironobu Koseki

    Full Text Available Biofilms forming on the surface of biomaterials can cause intractable implant-related infections. Bacterial adherence and early biofilm formation are influenced by the type of biomaterial used and the physical characteristics of implant surface. In this in vitro research, we evaluated the ability of Staphylococcus epidermidis, the main pathogen in implant-related infections, to form biofilms on the surface of the solid orthopaedic biomaterials, oxidized zirconium-niobium alloy, cobalt-chromium-molybdenum alloy (Co-Cr-Mo, titanium alloy (Ti-6Al-4V, commercially pure titanium (cp-Ti and stainless steel. A bacterial suspension of Staphylococcus epidermidis strain RP62A (ATCC35984 was added to the surface of specimens and incubated. The stained biofilms were imaged with a digital optical microscope and the biofilm coverage rate (BCR was calculated. The total amount of biofilm was determined with the crystal violet assay and the number of viable cells in the biofilm was counted using the plate count method. The BCR of all the biomaterials rose in proportion to culture duration. After culturing for 2-4 hours, the BCR was similar for all materials. However, after culturing for 6 hours, the BCR for Co-Cr-Mo alloy was significantly lower than for Ti-6Al-4V, cp-Ti and stainless steel (P0.05. These results suggest that surface properties, such as hydrophobicity or the low surface free energy of Co-Cr-Mo, may have some influence in inhibiting or delaying the two-dimensional expansion of biofilm on surfaces with a similar degree of smoothness.

  7. Effect of recombinant human growth hormone on early activities of myocardial myeloperoxidase in rats with scalded injury%重组人生长激素对严重烫伤大鼠早期心肌组织MPO活性的影响

    Institute of Scientific and Technical Information of China (English)

    朱国玲; 门秀丽; 樊学东; 刘朝永; 邱方

    2011-01-01

    目的 观察重组人生长激素(rhGH)对严重烫伤大鼠早期心肌组织髓过氧化物酶(MPO)活性的影响.方法 SPF级SD大鼠60只,雄性,随机分为对照组(B组,n=30),治疗组(R组,n=30).复制30%TBSAⅢ°烫伤模型,伤后B组按Parkland公式补液,R组在B组补液基础上于伤后6h始予rhGH 3U/(kg·d)皮下注射,两组均于烫伤后各时相点:0、12、24、48、96h检测血清心肌肌钙蛋白I(cTnI)含量和心肌组织MPO活性.结果 B组伤后不同时相点cTnI含量及MPO活性显著升高(P<0.01),伤后12h升高最为显著.心肌MPO活性升高与cTnI水平存在显著正相关(r=0.969,P<0.01).R组相应时相点cTnI含量及MPO活性与B组趋势一致,但较B组显著升高(P<0.05或0.01);病理切片显示:R组较B组炎细胞浸润、心肌损伤均有所加重.结论 严重烫伤后早期大鼠存在局灶性心肌炎,早期应用rhGH加重了烫伤后大鼠心肌损伤.%Objective To study the effect of recombinant human growth hormone(rhGH) on early activities of myocardial myeloperoxidase in rats with scald. Methods Sixty male Sprague-Dawley rats were randomly divided into control group and treatment group, 30 in each group. A 30% TBSA Ⅲ scald model was established. Rats in control group received fluid infusion after scald according to the formula of Parkland, and those in treatment group received fluid infusion with rhGH(3U/kg ? D) subcutaneously injected 6h after scald. Serum cTnI level and myocardial myeloperoxidase activities in both groups were measured at 0,12, 24,48, and 96h after scald. Results The serum cTnI level and myocardial myeloperoxidase activities were significantly higher in control group B at different time points after scald, especially 12h after scald(r=0.969, P<0.01). The same trends in serum cTnl level and myocardial myeloperoxidase activites was observed in treatment group. However, they were significantly higher than those in control group(P<0.05). Pathological examination showed

  8. 30例新生儿葡萄球菌皮肤烫伤综合征误诊分析%Misdiagnostic Analysis on 30 Cases with Neonatal Staphylococcus Aureus Scalded Skin Syndrome

    Institute of Scientific and Technical Information of China (English)

    邱梅冰; 沈琪

    2015-01-01

    Objective To analyse the situation of misdiagnosis of neonatal staphylococcal scalded skin syndrome( SSSS) ,and improve the understanding of neonatal SSSS,reduce the misdiagnosis rate. Methods The clinical data of 30 cases of misdiagnosis of neonatal were analysed.Results In 30 cases of misdiagnosis of neonatal SSSS,9 cases misdiagnosed as impetigo,7 cases misdiagnosed as al ergic dermatitis,6 cases misdiagnosed as drug eruption, 5 cases were misdiagnosed as eczema,2 cases misdiagnosed as burns,1 case was misdiagnosed as toxic epidermolysis disease. Conclusion To improve neonatal SSSS early identification, accumulating and sum up experience lesson, is expected to reduce the misdiagnosis rate.%目的分析新生儿葡萄球菌皮肤烫伤综合征(staphylococcal scalded skin syndrome,s s )误诊情况,提高对新生儿s s 的认识,减少误诊率。方法对30例误诊的新生儿s s 的临床资料进行分析。结果30例误诊的新生儿s s 中,9例误诊为脓疱疮,7例误诊为过敏性皮炎,6例误诊为药疹,5例误诊为湿疹,2例误诊为烫伤,1例误诊为中毒性表皮松解症。结论提高对新生儿ss 早期识别,不断积累教训和总结经验,可望减少误诊率。

  9. Developing a focused scald-prevention program.

    Science.gov (United States)

    Cagle, Kathleen M; Davis, James W; Dominic, William; Ebright, Steffanie; Gonzales, Wendy

    2006-01-01

    Scalds account for a high percentage of burn injuries in young children. The purpose of this project was to use a formative evaluation process to design a pilot scald-prevention program for a high-risk population. The burn registry and U.S. Census were used to define a high-risk population. A total of 53 children younger than age 6 were admitted to a local burn center with scalds during a 4-year period. Cooking or food accounted for 84% of these injuries. A total of 21% of the patients resided in one zip code, representing an incidence rate of 23 per 100,000, which was statistically significant. Focus group meetings were conducted with parents in this zip code. They were queried about scald injury knowledge, prevention practices, and attitudes toward interventions. A prevention program was designed based on the findings. Workshops are conducted with high-risk groups in the zip code. Attendees consent to a home visit where prevention practices are assessed and taught. A pre/post test and home risk assessment survey is used to measure change. The Burn Registry, U.S. Census, and focus groups were complimentary formative evaluation measures that assisted in developing a targeted scald prevention project.

  10. Programming MapReduce with Scalding

    CERN Document Server

    Chalkiopoulos, Antonios

    2014-01-01

    This book is an easy-to-understand, practical guide to designing, testing, and implementing complex MapReduce applications in Scala using the Scalding framework. It is packed with examples featuring log-processing, ad-targeting, and machine learning. This book is for developers who are willing to discover how to effectively develop MapReduce applications. Prior knowledge of Hadoop or Scala is not required; however, investing some time on those topics would certainly be beneficial.

  11. Patterns of scald injuries in children--has anything changed?

    LENUS (Irish Health Repository)

    Yates, J

    2011-10-01

    The objective was to study presentation patterns of scald injuries in children and suggest potential countermeasures to reduce these injuries. We retrospectively studied scald injuries in children attending an urban paediatric emergency department between January 1st and December 31st 2008. Data was extracted from our emergency department database using search terms \\'burn\\

  12. Clinical Observation of MEBO Combined with Early Functional Exercise in the Treatment of Burns and Scalds at Functional Sites%湿润烧伤膏联合早期功能锻炼治疗功能部位烧烫伤的临床观察

    Institute of Scientific and Technical Information of China (English)

    郑学勤; 常飞; 王淑文; 于丽琴

    2013-01-01

    Objective To observe the clinical curative effect of MEBO combined with early functional exercise in the treatment of burns and scalds at functional sites. Methods Forty-eight patients with burns and scalds were treated with MEBO externally and bandage therapy ( with dressing change every 6-8 hours) at the meantime of receiving anti-infection, anti-shock and nutritional support treatments. Meanwhile, the involved 88 functional sites were given early functional exerci-ses. The therapeutic effect was observed. Results All 48 cases were healed with a total effective rate of 100%. No com-plications such as local or systemic infection, high fever, severe malnutrition, scar formation, contracture malformation and dysfunction and etc. developed. During the follow-up of 3-6 months, no obvious scar hyperplasia, contracture or dysfunc-tion was observed. Conclusion MEBO combined with early functional exercise in the treatment of burns and scalds can not only promote wound healing, but also effectively prevent the occurrences of scar formation, contracture deformity and loss of function at functional sites.%  目的 观察湿润烧伤膏( MEBO)配合早期功能锻炼治疗功能部位烧烫伤的临床疗效。方法 对48例不同烧烫伤患者的88个功能部位,在给予抗感染、抗休克、营养支持等治疗的基础上,烧烫伤创面用MEBO涂抹、无菌纱布覆盖处理(每6 h~8 h换药1次),并早期进行功能部位的功能锻炼,观察疗效。结果 48例患者的创面全部愈合,总有效率为100%;所有患者均未出现全身或局部的严重感染、高热、营养不良、瘢痕形成、挛缩畸形、功能障碍等并发症,随访3~6个月,未见明显瘢痕增生、挛缩畸形及功能障碍。结论 湿润烧伤膏配合早期功能锻炼治疗烧烫伤,既可促进创面愈合,又可有效预防烧伤部位的瘢痕形成、挛缩畸形、功能丧失。

  13. Effect of Ampelopsis Radix on wound healing in scalded rats.

    Science.gov (United States)

    Lee, Kyungjin; Lee, Byonghee; Lee, Mi-Hwa; Kim, Bumjung; Chinannai, Khanita Suman; Ham, Inhye; Choi, Ho-Young

    2015-07-08

    Ampelopsis Radix has been used as a traditional Korean medicine for the treatment of burns and scalds. However, there has been no scientific research to date on the wound healing properties of Ampelopsis Radix for scald burns. This study aimed to evaluate the healing effect of Ampelopsis japonica root tuber ethanol extract (AJE) on induced cutaneous scald injury in Sprague Dawley (SD) rats. Hot water scalds were induced in SD rats, who were then divided into the following 5 groups; 1) control group without treatment, 2) positive control group with 1% Silver sulfadiazine (SSD), 3) Vaseline group, and groups 4) and 5) that used Vaseline containing 5% and 20% AJE, respectively. The ointment was applied topically to the experimental rats, once daily for 21 days, starting at 24 h post induction of the scald injury. Gross examination, measurement of wound size, and histopathological examination were performed. And quantitative measurement of cytokine levels of tumor necrosis factor alpha (TNF-α), interleukin-10 (IL-10), transforming growth factor beta 1 (TGF-β1), and vascular endothelial growth factor (VEGF) were performed by enzyme-linked immunosorbent assay. Clinical evaluation showed that the AJE and Vaseline groups, rapidly desquamated scab on day 12 post-scalding; in particular, the 20% AJE group achieved the greatest extent of skin recovery. Sizes of scald wound were significantly lower on days 12, 15, 18, and 21 in the AJE treated groups compared to the control groups. Histopathological evaluation showed a well-organized epithelial layer, angiogenesis, tissue granulation and collagen formation with the exception of inflammatory cells in the AJE-treated groups compared to the control groups on day 14, indicating that tissue regeneration had occurred. AJE treatment decreased TNF-α and increased IL-10 levels on days 2 and 14, indicating the anti-inflammatory action of AJE. The AJE groups also showed a decrease in TGF-β1 levels on day 7 and VEGF on day 14 in the

  14. Staphylococcal Toxic Shock Syndrome Caused by Tampon Use

    Directory of Open Access Journals (Sweden)

    Cian McDermott

    2015-01-01

    Full Text Available The authors report a case of near-fatal sepsis with multiorgan failure resulting from a Staphylococcal tampon-associated toxic shock syndrome, requiring a lengthy critical care admission. Successful treatment of this condition focuses on early identification, source control, and administration of antimicrobial agents. Intravenous immunoglobulin therapy used early may prevent widespread tissue necrosis.

  15. A case series of thermal scald injuries in dogs exposed to hot water from garden hoses (garden hose scalding syndrome).

    Science.gov (United States)

    Quist, Erin M; Tanabe, Mika; Mansell, Joanne E K L; Edwards, Jeffrey L

    2012-04-01

    In this report, we present a series of cases of thermal burns (scalds) in dogs resulting from exposure to hot water from a garden hose that had been lying in the sun. These dogs typically inhabited the southern and western regions of the USA, where the recorded high temperatures often exceed 32°C (90°F) during the warm summer months. Dogs with thermal scald injury in these cases presented with linear thermal burns along the dorsum, in addition to a variety of other macroscopic lesions that were dependent upon the degree of burn exposure and ranged from local erythema to ulcerated, necrotic and sloughing skin. Chronic, healed wounds were often alopecic, with markedly thickened skin and characteristically smooth and glassy scar tissue formation. Histologically, the lesions of thermal scald injury in these dogs were indistinguishable from any other second or third degree burn, and consisted of full-thickness dermal and epidermal necrosis with occasional fibrinoid necrosis of vessel walls, vasculitis and intravascular thrombosis. Here, we closely examine 10 cases of dogs with dorsal thermal burns collected from Texas, Arizona, California, Utah, Nevada, Indiana, Michigan and North Carolina and propose the term 'garden hose scalding syndrome (GHS)' to describe this unique type of scald injury.

  16. Results of a focused scald-prevention program.

    Science.gov (United States)

    Cagle, Kathleen M; Davis, James W; Dominic, William; Gonzales, Wendy

    2006-01-01

    Scalds are a leading cause of burn injury for young children. A focused prevention program was developed in the zip code accounting for the majority of scald burns. This study investigated the effect of the program. Families in the high-risk area were identified at clinics, community centers, and schools. Parent workshops and home visits were the interventions used. A pretest was administered at the workshop to measure baseline knowledge. A post-test was administered at either the home visit or by telephone to measure change in knowledge. A survey was used to measure baseline scald risks in the home. Home visits were used to reinforce information from workshops, evaluate the home environment, and assist parents to make environmental changes. Changes to the home environments were made, with antiscald devices installed in the shower, sink, or bathtub depending on parent preference. The survey was repeated on a follow-up home visit to determine whether parents adhered to environmental changes and safety practices. The postmeasurements were performed from 6 to 12 months after the initial measurement. More than 900 parents attended the initial workshops, and 173 consented to participate in the follow-up study and took the pretest. Of these, 62 completed the post-test, and 48 participated in a home visit. The mean pretest score was 72 +/- 1%, and mean post-test score was 85 +/- 1% (P burns in the homes in which the focused prevention program took place. This study demonstrates that a focused burn-prevention program can identify high-risk groups, decrease the number of scald risks per home, and decrease the rate of scald burns in the population. This straightforward program could be used to intervene in high-risk groups in other communities.

  17. Are We Serving Tea or Scalding Our Children?

    Directory of Open Access Journals (Sweden)

    Ilhan Ciftci

    2013-02-01

    Method: We retrospectively studied 255 patients between 0 and 5 years of age who suffered from major burns and who received inpatient treatment in the Burn Unit based on the guidelines of the American Burn Association between 2009 and 2011. The patients' age, gender, burn location, cause of burn, time to reach a health center after burn, and depth and width of the burn were evaluated. The possibility of a seasonal correlation was investigated as well. Results: Of the patients, 38.4% were female and 61.6% were male. The mean age of the patients was 2.1+/-1.2 years. The mean burned surface area amounted to 10.8% +/-7.4%. The nature of the burns was as follows: scalding in 73.7% (tea in 46.7%, hot water in 25.9%, and vapor in 1.2%, chemical burns in 16.1%, contact burns from hot contact in 5.5%, and flame burns in 4.7%. Conclusion: Most of the burns observed among preschool children occur indoors, with the majority of these burns represented by scalding burns. Most scalding burns arise from splashes of hot tea. In fact, burns associated with hot tea comprise nearly 50% of all burns. [Arch Clin Exp Surg 2013; 2(1.000: 32-37

  18. Effects of scalding method and sequential tanks on broiler processing wastewater loadings

    Science.gov (United States)

    The effects of scalding time and temperature, and sequential scalding tanks was evaluated based on impact to poultry processing wastewater (PPW) stream loading rates following the slaughter of commercially raised broilers. On 3 separate weeks (trials), broilers were obtained following feed withdrawa...

  19. Gene expression and metabolism preceding soft scald, a chilling injury of 'Honeycrisp' apple fruit.

    Science.gov (United States)

    Leisso, Rachel S; Gapper, Nigel E; Mattheis, James P; Sullivan, Nathanael L; Watkins, Christopher B; Giovannoni, James J; Schaffer, Robert J; Johnston, Jason W; Hanrahan, Ines; Hertog, Maarten L A T M; Nicolaï, Bart M; Rudell, David R

    2016-10-12

    'Honeycrisp' is an apple cultivar that is susceptible to soft scald, a chilling injury expressed as necrotic patches on the peel. Improved understanding of metabolism associated with the disorder would improve our understanding of soft scald and contribute to developing more effective management strategies for apple storage. It was expected that specific gene expression and specific metabolite levels in the peel would be linked with soft scald risk at harvest and/or specific time points during cold storage. Fruit from nine 'Honeycrisp' apple orchards that would eventually develop different incidences of soft scald between 4 and 8 weeks of cold air storage were used to contrast and determine differential transcriptomic and metabolomic changes during storage. Untargeted metabolic profiling revealed changes in a number of distinct pathways preceding and concurrent with soft scald symptom development, including elevated γ-aminobutryic acid (GABA), 1-hexanol, acylated steryl glycosides, and free p-coumaryl acyl esters. At harvest, levels of sesquiterpenoid and triterpenoid acyl esters were relatively higher in peel of fruit that did not later develop the disorder. RNA-seq driven gene expression profiling highlighted possible involvement of genes and associated metabolic processes with soft scald development. These included elevated expression of genes involved in lipid peroxidation and phenolic metabolism in fruit with soft scald, and isoprenoid/brassinosteroid metabolism in fruit that did not develop soft scald. Expression of other stress-related genes in fruit that developed soft scald included chlorophyll catabolism, cell wall loosening, and lipid transport while superoxide dismutases were up-regulated in fruit that did not develop the disorder. This study delineates the sequential transcriptomic and metabolomic changes preceding soft scald symptom development. Changes were differential depending on susceptibility of fruit to the disorder and could be attributed to

  20. Removal of amino-terminal extracellular domains of desmoglein 1 by staphylococcal exfoliative toxin is sufficient to initiate epidermal blister formation.

    Science.gov (United States)

    Nishifuji, Koji; Shimizu, Atsushi; Ishiko, Akira; Iwasaki, Toshiroh; Amagai, Masayuki

    2010-09-01

    In both bullous impetigo and staphylococcal scalded-skin syndrome (SSSS), exfoliative toxins (ETs) produced by Staphylococcus aureus cause superficial intraepidermal blisters. ETs are known to cleave specifically a single peptide bond in the extracellular domains 3 and 4 of desmoglein (Dsg) 1. However, the precise mechanisms underlying ET-induced epidermal blister formation remain poorly understood. To determine whether cleavage of Dsg1 by an ET is sufficient to induce blister formation in vivo or if the subsequent internalization of cleaved Dsg1 or other desmosomal components is required. Skin samples obtained from neonatal mice injected with ETA were analyzed by time-lapse immunofluorescence and transmission electron microscopy for desmosomal components. Epidermal blister formation was observed as early as 60 min after ETA treatment. At this time, the amino-terminal extracellular domains of Dsg1 disappeared from the surface of keratinocytes, while the cleaved carboxy-terminal domain of Dsg1 (Dsg1-C) as well as the extracellular domains of desmocollin 1 (Dsc1-N) remained on the cell surface. Half-split desmosomes with intracytoplasmic dense plaques and attached tonofilaments were recognized ultrastructurally on the split surface of keratinocytes at 60 min. Subsequent to this, Dsg1-C and Dsc1-N gradually disappeared from the surface layer of keratinocytes. Our findings suggest that the removal of amino-terminal extracellular domains of Dsg1 by ETs is sufficient to initiate epidermal blister formation in bullous impetigo and SSSS. Copyright 2010 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

  1. Electroacupuncture at Zusanli Prevents Severe Scalds-Induced Gut Ischemia and Paralysis by Activating the Cholinergic Pathway

    Directory of Open Access Journals (Sweden)

    Huan Wang

    2015-01-01

    Full Text Available Severe burn injuries may result in gastrointestinal paralysis, and barrier dysfunction due to gut ischemia and lowered vagus excitability. In this study we investigate whether electroacupuncture (EA at Zusanli (ST36 could prevent severe scalds-induced gut ischemia, paralysis, and barrier dysfunction and whether the protective role of EA at ST36 is related to the vagus nerve. 35% burn area rats were divided into six groups: (a EAN: EA nonchannel acupoints followed by scald injury; (b EA: EA at ST36 after scald injury; (c VGX/EA: vagotomy (VGX before EA at ST36 and scald injury; (d VGX/EAN: VGX before EAN and scald injury; (e atropine/EA: applying atropine before scald injury and then EA at ST36; (f atropine/EAN: applying atropine before scald injury and then EA at nonchannel acupoints. EA at the Zusanli point significantly promoted the intestinal impelling ratio and increased the amount of mucosal blood flow after scald injury. The plasma diamine oxidase (DAO and intestinal permeability decreased significantly after scald injury in the EA group compared with others. However, EA after atropine injection or cervical vagotomy failed to improve intestinal motility and mucosa blood flow suggesting that the mechanism of EA may be related to the activation of the cholinergic nerve pathway.

  2. Keeping children safe at home: protocol for a case–control study of modifiable risk factors for scalds

    Science.gov (United States)

    Wynn, P; Stewart, J; Kumar, A; Clacy, R; Coffey, F; Cooper, N; Coupland, C; Deave, T; Hayes, M; McColl, E; Reading, R; Sutton, A; Watson, M; Kendrick, D

    2014-01-01

    Background Scalds are one of the most common forms of thermal injury in young children worldwide. Childhood scald injuries, which mostly occur in the home, result in substantial health service use and considerable morbidity and mortality. There is little research on effective interventions to prevent scald injuries in young children. Objectives To determine the relationship between a range of modifiable risk factors for medically attended scalds in children under the age of 5 years. Design A multicentre case-control study in UK hospitals and minor injury units with parallel home observation to validate parental reported exposures. Cases will be 0–4 years old with a medically attended scald injury which occurred in their home or garden, matched on gender and age with community controls. An additional control group will comprise unmatched hospital controls drawn from children aged 0–4 years attending the same hospitals and minor injury units for other types of injury. Conditional logistic regression will be used for the analysis of cases and matched controls, and unconditional logistic regression for the analysis of cases and unmatched controls to estimate ORs and 95% CI, adjusted and unadjusted for confounding variables. Main exposure measures Use of safety equipment and safety practices for scald prevention and scald hazards. Discussion This large case-control study will investigate modifiable risk factors for scalds injuries, adjust for potential confounders and validate measures of exposure. Its findings will enhance the evidence base for prevention of scalds injuries in young children. PMID:24842981

  3. Staphylococcal toxic shock syndrome presenting as acute respiratory distress and cor pulmonale.

    Science.gov (United States)

    Zaki, S A; Shanbag, P; Chavan, V; Shenoy, P

    2010-01-01

    We describe a 7-year-old boy with staphylococcal toxic shock syndrome who presented with acute respiratory distress and cor pulmonale. We wish to highlight this unusual presentation as the diagnosis of toxic shock syndrome depends chiefly on a high degree of clinical suspicion. Early diagnosis and prompt institution of appropriate therapy will significantly reduce morbidity and mortality.

  4. 21 CFR 866.2050 - Staphylococcal typing bacteriophage.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Staphylococcal typing bacteriophage. 866.2050 Section 866.2050 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... Staphylococcal typing bacteriophage. (a) Identification. A staphylococcal typing bacteriophage is a...

  5. Wound Healing Effects of Prunus yedoensis Matsumura Bark in Scalded Rats

    Directory of Open Access Journals (Sweden)

    Jin-Ho Lee

    2017-01-01

    Full Text Available Pruni Cortex has been used to treat asthma, measles, cough, urticaria, pruritus, and dermatitis in traditional Korean medicine. The objective of this study was to investigate the effects of Prunus yedoensis Matsumura bark methanol extract (PYE on scald-induced dorsal skin wounds in rats. Scalds were produced in Sprague-Dawley rats with 100°C water and treated with 5% and 20% PYE (using Vaseline as a base, silver sulfadiazine (SSD, and Vaseline once a day for 21 days, beginning 24 hours after scald by treatment group allocation. The PYE-treated groups showed accelerated healing from 12 days after scald, demonstrated by rapid eschar exfoliation compared to the control and SSD groups. PYE-treated groups showed higher wound contraction rates and better tissue regeneration in comparison with the control group. Serum analysis showed that transforming growth factor beta 1 and vascular endothelial growth factor levels remained high or gradually increased up to day 14 in both PYE groups and then showed a sharp decline by day 21, implying successful completion of the inflammatory phase and initiation of tissue regeneration. These findings suggested that PYE is effective in promoting scald wound healing in the inflammation and tissue proliferation stages.

  6. FDA-approved immunosuppressants targeting staphylococcal superantigens: mechanisms and insights

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    Krakauer T

    2017-05-01

    Full Text Available Teresa Krakauer Department of Immunology, Molecular Translational Sciences Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, USA Abstract: Immunostimulating staphylococcal enterotoxin B (SEB and related superantigenic toxins cause diseases in human beings and laboratory animals by hyperactivating cells of the immune system. These protein toxins bind to the major histocompatibility complex class II (MHC II molecules and specific Vβ regions of T-cell receptors (TCRs, resulting in the stimulation of both monocytes/macrophages and T lymphocytes. The bridging of TCR with MHC II molecules by superantigens triggers intracellular signaling cascades, resulting in excessive release of proinflammatory mediators and massive polyclonal T-cell proliferation. The early induction of tumor necrosis factor α, interleukin 1 (IL-1, interleukin 2 (IL-2, interferon gamma (IFNγ, and macrophage chemoattractant protein 1 promotes fever, inflammation, and multiple organ injury. The signal transduction pathways for staphylococcal superantigen-induced toxicity downstream from TCR/major histocompatibility complex (MHC ligation and interaction of cell surface co-stimulatory molecules include the mitogen-activated protein kinase cascades and cytokine receptor signaling, activating nuclear factor kB (NFkB and the phosphoinositide 3-kinase/mammalian target of rapamycin pathways. Knowledge of host regulation within these activated pathways and molecules initiated by SEB and other superantigens enables the selection of US Food and Drug Administration (FDA-approved drugs to interrupt and prevent superantigen-induced shock in animal models. This review focuses on the use of FDA-approved immunosuppressants in targeting the signaling pathways induced by staphylococcal superantigens. Keywords: immunosuppressant, superantigen, toxic shock, NFkB, mTORC1

  7. Bacterial Toxins-Staphylococcal Enterotoxin B.

    Science.gov (United States)

    Fries, Bettina C; Varshney, Avanish K

    2013-12-01

    Staphylococcal enterotoxin B is one of the most potent bacterial superantigens that exerts profound toxic effects upon the immune system, leading to stimulation of cytokine release and inflammation. It is associated with food poisoning, nonmenstrual toxic shock, atopic dermatitis, asthma, and nasal polyps in humans. Currently, there is no treatment or vaccine available. Passive immunotherapy using monoclonal antibodies made in several different species has shown significant inhibition in in vitro studies and reduction in staphylococcal enterotoxin B-induced lethal shock in in vivo studies. This should encourage future endeavors to develop these antibodies as therapeutic reagents.

  8. Antibodies to staphylococcal enterotoxin in laboratory personnel.

    OpenAIRE

    Jozefczyk, Z; Robbins, R N; Spitz, J M; Bergdoll, M S

    1980-01-01

    Eighty-five percent of laboratory personnel working with staphylococcal enterotoxin had antibodies to enterotoxin in their sera, whereas only 23% of the control group had antibodies specific for enterotoxin. Two persons who carried enterotoxin B-producing staphylococci in their noses, throats, or both, had antibodies to enterotoxin B in their sera.

  9. Infantile chylothorax associated with staphylococcal paravertebral discitis

    Energy Technology Data Exchange (ETDEWEB)

    Ananthakrishnan, Ganapathy; Wilkinson, Alistair G.; McGurk, Simon F. [Royal Hospital for Sick Children, Department of Radiology, Edinburgh (United Kingdom); Marshall, Thomas [Royal Hospital for Sick Children, Department of Paediatrics, Edinburgh (United Kingdom)

    2009-12-15

    We report an unusual case of chylothorax in an infant associated with a staphylococcal paravertebral abscess secondary to discitis affecting the thoracic spine. We discuss the initial presentation with sepsis and chylothorax and the delayed diagnosis of discitis. We also discuss the imaging features and treatment in this case. (orig.)

  10. Adolescent scar contracture scoliosis caused by back scalding during the infantile period

    Science.gov (United States)

    Wang, S. F.; Wang, B.; Wu, L.; Zhu, F.

    2007-01-01

    The study design was a retrospective study in adolescent scar contracture scoliosis caused by back scalding during the infantile period. The objective of the study was to investigate the pathogenesis, clinical manifestation and treatment of adolescent scar contracture scoliosis caused by back scalding during the infantile period. This condition seldom occurs and is not reported in current English literature. One patient was first treated with skin expansion, back scar excision and skin flap transfer, followed by anterior correction with TSRH instrumentation. Two patients were first treated with back scar excision and anterior spinal release. One patient was treated with posterior correction with TSRH instrumentation, and thoracoplasty was performed after 50 days in halo-wheelchair traction. The other patient was treated with posterior correction with TSRH instrumentation. No management of scalding was performed on the fourth patient. Anterior release and posterior correction were performed at an interval of 3 weeks. The deformities of four patients were well corrected. Trunk balance was restored and the pelvis leveled. The skin incision wounds healed well. Minor loss of correction was recorded during the last follow-up. Severe scar contracture caused by back scalding during the infantile period could lead to adolescent scoliosis. Its pathogenesis and clinical manifestation are different from the typical adolescent idiopathic scoliosis. The treatment of this kind of scoliosis should be individualized. PMID:17497186

  11. A special type of scald caused by prolonged exposure to slowly heated water.

    Science.gov (United States)

    Pircher, R; Pollak, S; Vogt, S; Epting, T; Kramer, L; Geisenberger, D

    2016-06-01

    When skin is subjected to water temperatures gradually rising from moderate to critical ranges for a prolonged period of time, the resulting scalds will initially present as reddening, followed by blistering and at last by full-thickness burns. On changing from second- to third-degree burn, the blisters stop to become enlarged and solidify due to heat-induced coagulation necrosis. Such gradually intensifying tissue damage results in multiple firm skin blisters together with third-degree burn of the affected area. The initially serous blister fluid assumes a jelly-like consistency as the proteins undergo denaturation. Scalds of this special type may occur in persons taking a bath and becoming incapable to act so that the warm water tap remains turned on or a hairdryer continues to work after having been dropped into the bath water. On the basis of two relevant fatalities (suicidal electrocutions in bathtubs by means of hairdryers), the macroscopic and histological appearance of blisters which only secondarily turned into a full-thickness scald is compared with blisters in genuine second-degree scalds. The blister fluid of the latter ones was also investigated as to their protein composition and hemoglobin concentration. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  12. Effects of Extract of Dilong (Pheretima)on the Scalded Skin in Rats

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In order to investigate the effects of extract of Dilong (地龙 Pheretima) on the skin viability and lipid peroxidation after deep degree Ⅱ scalds, the extract of Dilong (Pheretima) at two concentrations (5%, 10%)were respectively applied to the surface of wound. The oxygen consumption, the succinate dehydrogenase (SDH) activity, the Schiff's base content, and the healing time of wound surface were determined in Wistar rats at the 8th, 24th, 48th and 72th hour after scalds, with SD-Ag used as controls. The results indicated that the oxygen consumption and SDH activity decreased, and the schiff's base content increased after the scalds. After the application of the extract of Dilong, the oxygen consumption and SDH activity increased, and the schiff's base content decreased significantly as compared with the control group (P<0.05 or P<0.01), and the mean healing time of the wound surface was five days in advance in the two treatment groups as compared with the control group. It is suggested that the scalds are related with lipid peroxdation to a certain extent, and the extract of Dilong has actions of attenuating lipid peroxidation, and promoting healing of the wound.

  13. The iron-regulated staphylococcal lipoproteins

    Directory of Open Access Journals (Sweden)

    Jessica eSheldon

    2012-04-01

    Full Text Available Lipoproteins fulfill diverse roles in antibiotic resistance, adhesion, protein secretion, signaling and sensing, and many also serve as the substrate binding protein (SBP partner to ABC transporters for the acquisition of a diverse array of nutrients including peptides, sugars, and scarcely abundant metals. In the staphylococci, the iron-regulated SBPs are significantly upregulated during iron starvation and function to sequester and deliver iron into the bacterial cell, enabling staphylococci to circumvent iron restriction imposed by the host environment. Accordingly, this subset of lipoproteins has been implicated in staphylococcal pathogenesis and virulence. Lipoproteins also activate the host innate immune response, triggered through Toll-like receptor-2 (TLR2 and, notably, the iron-regulated subset of lipoproteins are particularly immunogenic. In this review, we discuss the iron-regulated staphylococcal lipoproteins with regard to their biogenesis, substrate specificity, and impact on the host innate immune response.

  14. Bacterial Toxins—Staphylococcal Enterotoxin B

    OpenAIRE

    Fries, Bettina C.; Varshney, Avanish K.

    2013-01-01

    Staphylococcal enterotoxin B is one of the most potent bacterial superantigens that exerts profound toxic effects upon the immune system, leading to stimulation of cytokine release and inflammation. It is associated with food poisoning, nonmenstrual toxic shock, atopic dermatitis, asthma, and nasal polyps in humans. Currently, there is no treatment or vaccine available. Passive immunotherapy using monoclonal antibodies made in several different species has shown significant inhibition in in v...

  15. Staphylococcal food poisoning on a cruise ship.

    OpenAIRE

    Waterman, S H; Demarcus, T. A.; Wells, J G; Blake, P A

    1987-01-01

    Two waves of vomiting and/or diarrhoea affected approximately 215 of the 715 passengers on a Caribbean cruise ship. The outbreak was independently associated with eating cream-filled pastries at two separate meals. Staphylococcus aureus phage type 85/+ was isolated from cases and pastry cooks, but not from controls. This is the first well-documented outbreak of staphylococcal food poisoning on a cruise ship.

  16. Molecular Screening of Staphylococcal Enterotoxin B Gene in Clinical Isolates

    Directory of Open Access Journals (Sweden)

    Ali Karami

    2011-01-01

    Full Text Available Objective: The role of staphylococcal enterotoxin B (SEB in food poisoning is well known,however its role in other diseases remains to be explored. The aim of this study is the molecularscreening and characterization of the SEB gene in clinically isolated strains.Materials and Methods: In this experimentally study, 300 Staphylococcus aureus (S.aureus strains isolated from clinical samples were assayed. The isolated strains wereconfirmed by conventional bacteriological methods. Polymerase chain reaction (PCRwas used to determine the enterotoxin B (ent B gene. Assessment of toxin productionin all strains that contained the ent B gene was then performed. Finally, using specificantibody against SEB, a Western-blot was applied to confirm detection of enterotoxin Bproduction.Results: Results indicated that only 5% of the 300 clinically isolated S. aureus containedthe ent B gene. All strains which contained the ent B gene produced a proteinous enterotoxinB. The results of sequence determination of the PCR product were compared withthe gene bank database and 98% similarity was achieved. The results of the Western-blotconfirmed that enterotoxin B was produced in strains that contained the ent B gene.Conclusion: The results of this study indicate that 5% of clinically isolated S. aureusstrains produce enterotoxin B. Considering that the enterotoxin B is an important superantigen,it is possible that a delay in diagnosis and lack of early proper treatment can causean incidence of late complications, particularly in staphylococcal chronic infections. Forthis reason, it is suggested that in addition to detecting bacteria, an enterotoxin B detectiontest should be performed to control its toxigenicity.

  17. Scald-Induced Necrobiosis Lipoidica in a Patient with Diabetes Mellitus and Psoriasis

    Directory of Open Access Journals (Sweden)

    Hoshiko Ito

    2016-01-01

    Full Text Available The Koebner phenomenon (KP was first introduced by Heinrich Koebner in the 1870s to describe the appearance of psoriatic lesions following trauma in psoriasis patients. KP has since been defined in numerous diseases, including necrobiosis lipoidica diabeticorum (NLD. Since most Koebnerized dermatological lesions can localise to a site of previous trauma, Weiss et al. (Eur Acad Dermatol Venereol 2002;16:241-248 classified them into four categories (I-IV according to the Boyd-Nelder classification (Int J Dermatol 1990;29:401-410 system. In this system, necrobiosis lipoidica is classified as category III, which includes diseases that occasionally localise at the site of trauma. We report a case of NLD that developed after scald in a psoriasis patient. NLD after trauma has often been reported, but this is the first case of NLD that coincidentally occurred at a scald site in a psoriasis patient.

  18. Scald-Induced Necrobiosis Lipoidica in a Patient with Diabetes Mellitus and Psoriasis

    Science.gov (United States)

    Ito, Hoshiko; Imamura, Sadao

    2016-01-01

    The Koebner phenomenon (KP) was first introduced by Heinrich Koebner in the 1870s to describe the appearance of psoriatic lesions following trauma in psoriasis patients. KP has since been defined in numerous diseases, including necrobiosis lipoidica diabeticorum (NLD). Since most Koebnerized dermatological lesions can localise to a site of previous trauma, Weiss et al. (Eur Acad Dermatol Venereol 2002;16:241–248) classified them into four categories (I–IV) according to the Boyd-Nelder classification (Int J Dermatol 1990;29:401–410) system. In this system, necrobiosis lipoidica is classified as category III, which includes diseases that occasionally localise at the site of trauma. We report a case of NLD that developed after scald in a psoriasis patient. NLD after trauma has often been reported, but this is the first case of NLD that coincidentally occurred at a scald site in a psoriasis patient. PMID:26889139

  19. Life threatening deep scald burns in a neonate: A rare case report

    Directory of Open Access Journals (Sweden)

    Vijay Y Bhatia

    2013-01-01

    Full Text Available Domestic burns in neonate are rare in literature. Neonates are high risk for burns because of their thin and fragile skin, fluid overload or dehydration as the fluid balance range is small and immature immune system leading to septicaemia. Neonates are not small adults, owing to their different physiological response makes the management of neonates challenging as the clinical signs are different from the adults and the resuscitation protocols or end points are also different. We present a case of 11 days old neonate, who sustained 51% scald burns when the hot water bag being used by her mother accidentally burst, the youngest reported case of domestic scald burns with such high percentage. The baby was managed by fluid resuscitation, antibiotics, dressings, ventilatory support for septicaemia and subsequently homograft application. The neonate was discharged with completely healed areas after 35 days of burns and is on regular follow up and no complications have been observed.

  20. Evolutionary Origin of the Staphylococcal Cassette Chromosome mec (SCCmec)

    DEFF Research Database (Denmark)

    Rolo, Joana; Worning, Peder; Nielsen, Jesper Boye

    2017-01-01

    Several lines of evidence indicate that the most primitive staphylococcal species, those of the Staphylococcus sciuri group, were involved in the first stages of evolution of the staphylococcal cassette chromosome mec (SCCmec), the genetic element carrying the β-lactam resistance gene mecA. Howev...

  1. Chromatofocusing in the purification of staphylococcal enterotoxin D.

    Science.gov (United States)

    Lei, Z; Reiser, R F; Bergdoll, M S

    1988-06-01

    A chromatofocusing procedure for the purification of staphylococcal enterotoxin D was developed. The purification included the removal of the toxic protein from culture supernatant fluids of Staphylococcus aureus 1151m by batch adsorption with CG-50 resin, chromatofocusing on Polybuffer Exchanger 94, and gel permeation chromatography on Sephacryl S-200. The purity of the staphylococcal enterotoxin D obtained was approximately 98%.

  2. Chromatofocusing in the purification of staphylococcal enterotoxin D.

    OpenAIRE

    Lei, Z.; Reiser, R F; Bergdoll, M S

    1988-01-01

    A chromatofocusing procedure for the purification of staphylococcal enterotoxin D was developed. The purification included the removal of the toxic protein from culture supernatant fluids of Staphylococcus aureus 1151m by batch adsorption with CG-50 resin, chromatofocusing on Polybuffer Exchanger 94, and gel permeation chromatography on Sephacryl S-200. The purity of the staphylococcal enterotoxin D obtained was approximately 98%.

  3. [Mortality factors in flame and scalds burns: our experience in 816 patients].

    Science.gov (United States)

    Al, Behçet; Yildirim, Cuma; Coban, Sacit; Aldemir, Mustafa; Güloğlu, Cahfer

    2009-11-01

    Our aim was to evaluate the effective factors on mortality in flame and scald burns in the Diyarbakir region. The data of 816 patients who applied to the Dicle University Emergency Department between January 2001 and May 2005 with flame and scald burns were investigated retrospectively. The patients were separated into two groups as alive or deceased. Gender, age, burns shapes, burn degrees and rates, burn regions, admission periods, hospitalization times, complications, and the treatments were analyzed. 43.5% of the patients were female and 57.5% were male. Six hundred fifty-eight patients were under 6 years old. 70.5% of burns occurred as a result of negligence; 76.5% occurred due to scald and 23.5% due to flame. In cases of death, 39 patients were under 10 years old. The mean age was 9.32 years. The average hospitalization period was 10.37 days. The most common complication was wound infection. The mortality rate was 6.1%. The mortality rate was higher among patients who were hospitalized longer than 15 days (p = 0.030); whose burns were due to suicide attempt (p = 0.002); who used shoe paint on the burn wounds instead of treatment (p = 0.000); who had more than 40% second-degree burns (p = 0.000) or more than 20% third-degree burns (p = 0.000); and among those with acute respiratory failure, compartment syndrome, hypoalbuminemia, and sepsis (p = 0.000).

  4. Evolutionary paths of streptococcal and staphylococcal superantigens

    Directory of Open Access Journals (Sweden)

    Okumura Kayo

    2012-08-01

    Full Text Available Abstract Background Streptococcus pyogenes (GAS harbors several superantigens (SAgs in the prophage region of its genome, although speG and smez are not located in this region. The diversity of SAgs is thought to arise during horizontal transfer, but their evolutionary pathways have not yet been determined. We recently completed sequencing the entire genome of S. dysgalactiae subsp. equisimilis (SDSE, the closest relative of GAS. Although speG is the only SAg gene of SDSE, speG was present in only 50% of clinical SDSE strains and smez in none. In this study, we analyzed the evolutionary paths of streptococcal and staphylococcal SAgs. Results We compared the sequences of the 12–60 kb speG regions of nine SDSE strains, five speG+ and four speG–. We found that the synteny of this region was highly conserved, whether or not the speG gene was present. Synteny analyses based on genome-wide comparisons of GAS and SDSE indicated that speG is the direct descendant of a common ancestor of streptococcal SAgs, whereas smez was deleted from SDSE after SDSE and GAS split from a common ancestor. Cumulative nucleotide skew analysis of SDSE genomes suggested that speG was located outside segments of steeper slopes than the stable region in the genome, whereas the region flanking smez was unstable, as expected from the results of GAS. We also detected a previously undescribed staphylococcal SAg gene, selW, and a staphylococcal SAg -like gene, ssl, in the core genomes of all Staphylococcus aureus strains sequenced. Amino acid substitution analyses, based on dN/dS window analysis of the products encoded by speG, selW and ssl suggested that all three genes have been subjected to strong positive selection. Evolutionary analysis based on the Bayesian Markov chain Monte Carlo method showed that each clade included at least one direct descendant. Conclusions Our findings reveal a plausible model for the comprehensive evolutionary pathway of streptococcal and

  5. Intranasal Rapamycin Rescues Mice from Staphylococcal Enterotoxin B-Induced Shock

    Science.gov (United States)

    2012-09-18

    Mice from Staphylococcal Enterotoxin B-Induced Shock Teresa Krakauer * and Marilyn Buckley Integrated Toxicology Division, U.S. Army Medical...2012 / Published: 18 September 2012 Abstract: Staphylococcal enterotoxin B (SEB) and related exotoxins produced by Staphylococcus aureus are...allowing a wider therapeutic window. Keywords: intranasal rapamycin; staphylococcal enterotoxin B; shock 1. Introduction Staphylococcal

  6. Binding of flavonoids to staphylococcal enterotoxin B.

    Science.gov (United States)

    Benedik, Evgen; Skrt, Mihaela; Podlipnik, Crtomir; Ulrih, Nataša Poklar

    2014-12-01

    Staphylococcal enterotoxins are metabolic products of Staphylococcus aureus that are responsible for the second-most-commonly reported type of food poisoning. Polyphenols are known to interact with proteins to form complexes, the properties of which depend on the structures of both the polyphenols and the protein. In the present study, we investigated the binding of four flavonoid polyphenols to Staphylococcal enterotoxin B (SEB) at pH 7.5 and 25 °C: (-)-epigallocatechin-3-gallate (EGCG), (-)-epigallocatechin (EGC), kaempferol-3-glucoside (KAM-G) and kaempferol (KAM). Fluorescence emission spectrometry and molecular docking were applied to compare experimentally determined binding parameters with molecular modeling. EGCG showed an order of magnitude higher binding constant (1.4 × 10(5) M(-1)) than the other studied polyphenols. Our blind-docking results showed that EGCG and similar polyphenolic ligands is likely to bind to the channel at the surface of SEB that is responsible for the recognition of the T-cell beta chain fragment and influence the adhesion of SEB to T cells.

  7. Mechanisms of staphylococcal enterotoxin-induced emesis.

    Science.gov (United States)

    Hu, Dong-Liang; Nakane, Akio

    2014-01-05

    Pathogenic bacteria use various strategies to interact with the host organisms. Among them, toxin production constitutes an efficient way to alter specific functions of target cells. Various enterotoxins interact with the enteric nervous system, by stimulating afferent neurons or inducing neurotransmitter release from enterochromaffin cells which result either in vomiting, diarrhea, or in the intestinal inflammation process. Staphylococcus aureus produces a wide variety of toxins including staphylococcal enterotoxins (SEs) with demonstrated emetic activity; and staphylococcal enterotoxin-like (SEl) proteins, which are not emetic in a primate model or have yet to be tested. SEs and SEls have been traditionally subdivided into classical (SEA to SEE) and new (SEG to SElX) types. These toxins possess superantigenic activity and are highly resistant to denaturation which allows them to remain intact in contaminated foods and trigger food poisoning outbreaks. Symptoms are of rapid onset, and include nausea and violent vomiting. SEA is the most recognizable toxin causing food poisoning in humans throughout the world. However, it remains unclear how SEs induce emesis and via which signal pathway. This review is divided into four parts, and will focus on the following: (1) how bacterial toxins interact with the nervous system, (2) biological characteristics of SEs and SEls, (3) mechanisms of SE-induced emesis, and (4) use of a vaccine for the prevention of SE-induced emesis.

  8. Staphylococcal nasal carriage of health care workers.

    Science.gov (United States)

    Akhtar, Naeem

    2010-07-01

    To determine the frequency of staphylococcal nasal carriage of health care workers (HCWs) and antimicrobial susceptibility profile of the isolates for appropriate decolonization therapy. An observational study. The study was conducted at Holy Family Hospital, Rawalpindi, during the period from May 2007 to April 2008. Nasal swabs from anterior nares of HCWs were cultured and identified as Staphylococcus aureus, coagulasenegative staphylococci (CoNS), methicillin-resistant S. aureus (MRSA), methicillin-resistant CoNS (MRCoNS) by using standard methods. Antimicrobial susceptibility testing was performed on Muller Hinton Agar using disc diffusion method. Of the 468 HCWs, 213 (45.5%) participants were men and 255 (54.5%) were women. Eighty five (18.2%) were nasal carriers of S. aureus, 07 (1.5%) for MRSA, 343 (73.3%) for CoNS and 10 (2.1%) for MRCoNS. The highest carriage rate for S. aureus was in midwives (30%) followed by maintenance staff (28.6%), security guards (25%), technicians (23.5%), staff nurses (22.7%) and 90% of S. aureus and CoNS were susceptible to amikacin, gentamicin and fluoroquinolones tested. Fluoroquinolones, preferably oral levofloxacin in combination with topical gentamicin ointment, in places like Pakistan where mupirocin is not routinely available, can be used for decolonization of nasal staphylococcal carriage.

  9. Engineering of staphylococcal surfaces for biotechnological applications.

    Science.gov (United States)

    Wernérus, Henrik; Lehtiö, Janne; Samuelson, Patrik; Ståhl, Stefan

    2002-06-13

    Novel surface proteins can be introduced onto bacterial cell surfaces by recombinant means. Here, we describe various applications of two such display systems for the food-grade bacteria Staphylococcus carnosus and Staphylococcus xylosus, respectively. The achievements in the use of such staphylococci as live bacterial vaccine delivery vehicles will be described. Co-display of proteins and peptides with adhesive properties to enable targeting of the bacteria, have significantly improved the vaccine delivery potential. Recently, protective immunity to respiratory syncytial virus (RSV) could be evoked in mice by intranasal immunization using such 'second generation' vaccine delivery systems. Furthermore, antibody fragments and other 'affinity proteins' with capacity to specifically bind a certain protein, e.g. Staphylococcus aureus protein A-based affibodies, have been surface-displayed on staphylococci as initial efforts to create whole-cell diagnostic devices. Surface display of metal-binding peptides, or protein domains into which metal binding properties has been engineered by combinatorial protein engineering, have been exploited to create staphylococcal bioadsorbents for potential environmental or biosensor applications. The use of these staphylococcal surface display systems as alternatives for display of large protein libraries and subsequent affinity selection of relevant binding proteins by fluorescence-activated cell sorting (FACS) will be discussed.

  10. 葡萄球菌皮肤烫伤样综合征的快速诊断%Rapid diagnosis of staphylococcal scalded skin syndrome

    Institute of Scientific and Technical Information of China (English)

    庞传超; 崔绍山; 王爽; 赵琦

    2003-01-01

    目的:早期准确地诊断葡萄球菌皮肤烫伤样综合征(SSSS)并使其得到及时治疗.方法:选取编码剥脱毒素的金葡菌基因eta和etb的特异核苷酸序列为引物,以SSSS患者原发皮损的疱液或结痂经裂解处理后为模板进行DNA扩增,与ELISA检测培养分离菌株的剥脱毒素的结果对比.结果:34例SSSS患者PCR阳性率eta 61.74%、etb 21.0%、eta和etb 11.76%;ELISA阳性率ETA 29.41%、ETB 11.76%、ETA和ETB 2.94%.经统计学处理差异有显著性(P<0.01), PCR方法较ELISA方法敏感、准确.结论:PCR技术对SSSS的早期诊断、及时治疗和降低死亡率有重要价值.

  11. An unsuspected cause of meal-time morbidity: instant noodle scald burns.

    Science.gov (United States)

    Koltz, Peter F; Wasicek, Philip; Mays, Chester; Bell, Derek E

    2013-01-01

    Observational analysis revealed a concerning frequency of scald burns secondary to instant noodles. A literature review reveals studies with small sample sizes of pediatric populations and analysis of container engineering. The adult cohort, treatments, and short-term outcomes have been neglected. Considering these deficiencies, we reviewed our institution's experience with burns secondary to instant noodles. Patient encounters due to instant noodle burns from January 1, 2007, through May 15, 2011, were reviewed. Demographics, burn characteristics, treatment, length of stay, number of operative interventions, and complications were analyzed. Eight hundred fifty-two patients were seen (460 were admitted) for scald burns of all pathogenesis. Of these, 121 (14%) were seen for burns secondary to noodles (63 men and 58 women). Of these, 48 were older than age 4 (group 1), and 73 were younger than age 4 (group 2). TBSA was 2.34 in group 1 and 1.64 in group 2 (P = .04). The most commonly burned areas in group 1 were extremities (n = 43) and in group 2 were chest (n = 32) and extremities (n = 31). Seven patients in group 1 and two patients in group 2 required operative intervention. Length of stay in groups 1 and 2 were 3.5 and 6 days, respectively. Noodle scald burns cause morbidity at all ages. Pediatric burns due to noodles are frequently managed conservatively but more often necessitate inpatient treatment. The nonpediatric population has larger TBSA and requires more frequent operative intervention. The morbidity of noodle burns is significant. Increased public education and container re-engineering is warranted.

  12. Improving the microbiological safety of veal: validation of hot water rinse plus lactic acid spray in combination with scalding for hide-on carcasses to control Shiga-toxin producing Escherichia coli (STEC)

    Science.gov (United States)

    Bob veal calves (n=6) were assigned to six hide-on carcass treatments for each of 5 replications. Primary treatments were no-scald, scald, or scalding with chemical; secondary treatments were no-treatment control or lactic acid. Hides were inoculated with non-pathogenic rifampicin-resistant Escheric...

  13. Epidemiologic study of scald burns in victims in Tehran burn hospital

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    Kamran Aghakhani

    2013-10-01

    Full Text Available Background: Damages caused by scald burns are common and can cause severe complications and death. The purpose of this study was to define risk groups and then methods of prevention and treatment is designed to fit. Methods: Data for this retrospective study of hospitalized patients in Shahid Motahari Hospital in Tehran from 2007-2011 were compiled. Data including age, sex, cause of burn, and degree of burn and ultimate fate of the victims were collected from scald burns. Burns caused by boiling water and hot food (Scald , in two age groups : 12 and under 12 years ( children and more than 12 years ( adults were compared in terms of statistics . Results: A total of 1150 patients consisting of males (57.9% and females (42.1% were studied. The most common age was 1 year old and 50% of patients were under 3 years of age. 87.9% burned with boiling water and 12.1% had experienced burns with hot food. Incentive to burn was 0.3% cross burning and 99.7% incident. A maximum number of burns in children 12 years and younger males (42.1% and a minimum number in men over 12 years (15.7% were observed. Mean percentage of burns was 11% in over 12 years group and 30.9% in 12 and under 12 years group. The average hospital stay was 11.4 days and the mortality rate was 4.8%. The final status of the patients was as fallows: full recovery 904 cases (78.6%, partial recovery 134 (11.7%, clearance with personal consent 41 (3.6%, death 55 (4.8% and 16 cases (3.1% were among other reasons. Conclusion: In general it can be said, scald burns incidence in individuals aged 12 and younger were more than the older ones and the mean of burns was lower in individuals with over 12 years old. There was a sexual preference for males under 12 years. Mortality rate in the two groups has not any statistically significant difference. There was no statistically significant association between sex and mortality rate. Some of our findings are depending on cultural, social and economic

  14. How Should Staphylococcal Food Poisoning Outbreaks Be Characterized?

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    Sylviane Dragacci

    2010-08-01

    Full Text Available Staphylococcal food poisoning is one of the most common food-borne diseases and results from the ingestion of staphylococcal enterotoxins (SEs preformed in food by enterotoxigenic strains of Staphylococcus aureus. To date, more than 20 SEs have been described: SEA to SElV. All SEs have superantigenic activity whereas only a few have been proved to be emetic, representing a potential hazard for consumers. Characterization of staphylococcal food poisoning outbreaks (SFPOs has considerably progressed compared to 80 years ago, when staphylococci were simply enumerated and only five enterotoxins were known for qualitative detection. Today, SFPOs can be characterized by a number of approaches, such as the identification of S. aureus biovars, PCR and RT-PCR methods to identify the se genes involved, immunodetection of specific SEs, and absolute quantification by mass spectrometry. An integrated gene-to-protein approach for characterizing staphylococcal food poisoning is advocated.

  15. Changes of biological functions of dipeptide transporter (PepT1)and hormonal regulation in severe scald rats

    Institute of Scientific and Technical Information of China (English)

    Bing-Wei Sun; Xiao-Chen Zhao; Guang-Ji Wang; Ning Li; Jie-Shou Li

    2003-01-01

    AIM: To determine the regulatory effects of recombinant human growth hormone (rhGH) on dipeptide transport (PepT1) in normal and severe scald rats.METHODS: Male Sprague-Dawley rats with 30 % total body surface area (TBSA) Ⅲ degree scald were employed as the model. In this study rhGH was used at the dose of 2 IU.kg-1d-1. An everted sleeve of intestine 4 cm long obtained from mid-jejunum was securely incubated in Kreb's solution with radioactive dipeptide (3H-glycylsarcosine, 3H-Gly-Sar,10 μCi/ml) at 37 ℃ for 15 min to measure the effects of uptake and transport of PepT1 of small intestinal epithelial cells in normal and severe scald rats.RESULTS: Abundant blood supply to intestine and mesentery was observed in normal and scald rats administered rhGH,while less supply of blood to intestine and mesentery was observed in rats without rhGH. Compared with controls, the transport of dipeptide in normal rats with injection of rhGH was not significantly increased (P=0.1926), while the uptake was significantly increased (P=0.0253). The effects of transport and uptake of PepT1 in scald rats with injection of rhGH were significantly increased (P=0.0082, 0.0391).CONCLUSION: Blood supply to intestine and mesentery of rats was increased following injection of rhGH. The effects of uptake and transport of dipeptide transporters in small intestinal epithelial cells of rats with severe scald were markedly up-regulated by rhGH.

  16. Nematode Infections Are Risk Factors for Staphylococcal Infection in Children

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    Sandra F Moreira-Silva

    2002-04-01

    Full Text Available Nematode infection may be a risk factor for pyogenic liver abscess in children and we hypothesized that the immunomodulation induced by those parasites would be a risk factor for any staphylococcal infection in children. The present study was designed to compare, within the same hospital, the frequency of intestinal nematodes and Toxocara infection in children with and without staphylococcal infections. From October 1997 to February 1998, 80 children with staphylococcal infection and 110 children with other diseases were submitted to fecal examination, serology for Toxocara sp., evaluation of plasma immunoglobulin levels, and eosinophil counts. Mean age, gender distribution, birthplace, and socioeconomic conditions did not differ significantly between the two groups. Frequency of intestinal nematodes and positive serology for Toxocara, were remarkably higher in children with staphylococcal infections than in the non-staphylococcal group. There was a significant correlation between intestinal nematodes or Toxocara infection and staphylococcal infection in children, reinforced by higher eosinophil counts and higher IgE levels in these children than in the control group. One possible explanation for this association would be the enhancement of bacterial infection by the immunomodulation induced by helminth infections, due to strong activation of the Th2 subset of lymphocytes by antigens from larvae and adult worms.

  17. Grain yield, net blotch and scald of barley in Finnish official variety trials

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    J. ROBINSON

    2008-12-01

    Full Text Available Data on grain yield, and terminal severity of net blotch (Pyrenophora teres f. teres and scald (Rhynchosporium secalis from Finnish official barley (Hordeum vulgare variety trials were analysed to indicate the pattern of disease incidence over six years and five sites for nineteen barley genotypes, and the effect of the diseases on yield and the genotype by environment interaction for yield. The effect of climatic factors on net blotch severity were also investigated. The genotype by site interaction for net blotch severity was not statistically significant, but that for yield was. Net blotch severity differed between years, but was similar across sites and there were statistically significant first order interactions between year, site and genotype. 'Saana' and 'Thule' had relatively low mean terminal net blotch scores and their reaction to the disease was less sensitive to the environment than was that of 'Tyra' for example. Analysis of yield data adjusted for net blotch severity indicated that the magnitude of the genotype by environment interaction terms were not accounted for to any significant degree by differences in relative net blotch resistances among the barley genotypes. Overall, mean scores for scald severity were lower than those for net blotch. Terminal net blotch severity was correlated with May rainfall and growing degree days.;

  18. Microbiology and evisceration efficiency of broiler carcasses slaughtered and held up to 8 hours postmortem prior to scalding and defeathering

    Science.gov (United States)

    The implementation of on farm slaughter could eliminate potential animal welfare issues associated with cooping, transport, dumping, and shackling live broilers. This research evaluated evisceration efficiency and the microbiological implications of delaying scalding and defeathering for up to 8 h a...

  19. A Community-Based Programme for the Prevention of Burns and Scalds to Pre-School Children.

    Science.gov (United States)

    Harre, Niki; Polzer-Debruyne, Andrea

    1998-01-01

    A New Zealand program was designed to reduce burn and scald hazards in the home. Seven groups of parents and caregivers of preschool children participated. Three months after the program, selected participants had sustained 68% of the positive practice changes they had made. Evaluation also noted barriers to participant change and strengths and…

  20. Effect of sunlight-exposure on antioxidants and antioxidant enzyme activities in 'd'Anjou' pear in relation to superficial scald development.

    Science.gov (United States)

    Zhao, Jing; Xie, Xingbin; Shen, Xiang; Wang, Yan

    2016-11-01

    Influence of preharvest sunlight exposure on superficial scald development in 'd'Anjou' pears during cold storage was investigated. The biochemical changes related to scald including α-farnesene, conjugated trienols (CTols), antioxidants, antioxidant enzyme activities were monitored among separated blushed and shaded peels of unbagged fruit as well as the whole peel of bagged fruit. In unbagged fruit, scald symptom was restricted to shaded peel; while there was no difference in α-farnesene between blushed and shaded peels, CTols increased significantly in shaded peel along with scald development after 3months storage. Bagging treatment increased both α-farnesene and CTols significantly and enhanced scald. Preharvest sunlight exposure significantly increased certain antioxidant contents and antioxidant enzyme activities in blushed peel at harvest and during storage. These results reveal a direct role of CTols during development of scald, however, antioxidant systems may play an important role in α-farnesene oxidation to CTols and scald susceptibility in 'd'Anjou pears. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Nosocomial Staphylococcal Toxic Shock. Case Report

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    Arbune Manuela

    2016-07-01

    Full Text Available Staphylococcal toxic shock syndrome (STSS is a rare, potentially lethal infection, with a clinical picture of multiple organ dysfunction and shock. The etiology is Staphylococcus aureus exotoxin, while enterotoxins act as superantigens. Most cases are identified in women using a vaginal tampon. A 51-year-old female, with a past medical history of biliary dyskinesia, presented in the emergency room complaining of sudden onset of abdominal pain, vomiting, headache, myalgia, and chills. The initial diagnosis was biliary colic and was treated parenterally with Amoxi-clavulanate and fluid replacement. Initially, progress was unsatisfactory. Four days after admission she developed a systemic inflammatory syndrome, diffuse rash, jaundice, oliguria, confusion, persistent hypotension and biological evidence of thrombocytopenia, nitrogen retention, and cholestasis. She was admitted to the intensive care unit. A gluteal phlegmon induced after intramuscular injections was identified and surgically treated. Blood bacteriological cultures were negative, though MRSA was isolated in phlegmon pus. A diagnosis of STSS was based on CDC criteria.

  2. Profilaxis, immunoprophylaxis and therapy of staphylococcal mastitis

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    Vakanjac Slobodanka

    2006-01-01

    Full Text Available Inflammation of the mammary gland, mastitis, in cows presents one of the most acute problems in intensive milk production, incurring huge economic losses which reach from 20-80% even in developed countries in the past decade. On the grounds of the programme of the respective Ministry, the Veterinary Service of the Republic of Serbia is obliged to monitor, detect, curb and control infective inflammation of the mammary gland caused by staphylococcus and streptococcus. Long-term different approaches to mastitis treatment did not result in the required solution, so that the problem of mastitis is still present and acute. In order to prevent the infiltration of the pathogenic cause into the mammary gland, its populating and multiplication, it is necessary to maintain constant and regular controls of milk, as well as undertake of preventive and therapeutic measures aimed at reducing the incidence of mastitis. Immunization and immunotherapy of mastitis are new and very interesting areas for scientific investigations and work. In the recent decades vaccines against staphylococcal mastitis are being successfully developed, whose success is reflected in the reduced incidence of clinical and subcilinical mastitis.

  3. Role of the Kidney in Staphylococcal Enterotoxemia

    Science.gov (United States)

    Staab, Edward V.; Niederhuber, John; Rhoda, David A.; Faulkner, Charles S.; Beisel, William R.

    1969-01-01

    Highly purified staphylococcal enterotoxin B (SEB) is known to accumulate rapidly within the kidneys of experimental animals. The present study was performed to determine whether the predominant renal localization of SEB was of fundamental pathophysiologic importance in the development of lethal shock after the intravenous administration of this toxin to monkeys. Eight bilaterally nephrectomized Macaca mulatta given 10 μg of SEB per kg survived for an average time period less than half that of nephrectomized control animals (P < 0.001). Their survival time, however, was similar to that of control, sham-nephrectomized monkeys given an equal amount of SEB. Thus, no evidence was obtained to suggest that the kidney converted purified SEB to a more potent toxin. The glomerular filtration and proximal tubule cell accumulation of SEB possibly occurred as a nonspecific consequence of its molecular size, and such localization within the kidney might have served to reduce the quantity of SEB reaching some other site of toxic activity. Similar pathological and clinical findings were demonstrated in monkeys from both experimental and control groups; these could not be ascribed to SEB alone. PMID:4976324

  4. Effect of DPA and 1-MCP on chemical compounds related to superficial scald of Granny Smith apples

    Energy Technology Data Exchange (ETDEWEB)

    Moggia, C.; Moya-Leon, M. A.; Pereira, M.; Yuri, J. A.; Lobos, G. A.

    2010-07-01

    Research was carried out to study the mode of action of diphenylamine (DPA) and 1-methylcyclopropene (1-MCP), on control of superficial scald of Granny Smith apples (Malus domestica Borkh.), and its relation with chemical compounds. Fruit was harvested from a commercial orchard in Chile, 182 and 189 days after full bloom and received the following treatments: DPA (2,000 ppm); 1-MCP (1.2 ppm) and control (no treatment). All fruit was stored for 4 or 6 months at 0 degree centigrade. A completely randomized factorial design was used (2 harvest dates by 3 post harvest treatments). Monthly measurements were made on maturity indices, ethylene production rate (EPR), scald related compounds [a-farnesene (AF), conjugated trienes (CT), total anti-oxidants (AO)], and cell membrane stability. Following 4 and 6 months of storage, plus 7 days at 20 degree centigrade, scald was evaluated. After 6 months, DPA-treated fruit, from both harvests, showed similar firmness, EPR and AO, compared to the control. However, AF and CT were lower, and cell membrane stability higher. Conversely, 1-MCP-treated fruit showed a noticeable EPR suppression and AF inhibition, along with higher firmness, lower CT and AO, compared to the control and DPA. Furthermore, cell membrane stability was superior to that of the control and similar to that of the DPA. Treated fruit (DPA and 1-MCP) showed an important reduction in scald compared to the control. The effect of 1-MCP on the investigated compounds and the reduction in scald, confirms that ethylene plays a major role on its development. (Author) 50 refs.

  5. Phylogeographical analyses reveal global migration patterns of the barley scald pathogen Rhynchosporium secalis.

    Science.gov (United States)

    Zaffarano, Pascal L; McDonald, Bruce A; Linde, Celeste C

    2009-01-01

    A phylogeographical analysis of the scald pathogen Rhynchosporium secalis was conducted using nuclear DNA sequences from two neutral restriction fragment length polymorphism loci and the mating-type idiomorphs. Approximately 500 isolates sampled from more than 60 field populations from five continents were analysed to infer migration patterns and the demographic history of the fungus. Migration rates among continents were generally low, consistent with earlier reports of significant population subdivision among continents. Northern Europe was mainly a source population for global migration. We hypothesize that the pathogen only recently moved out of its centre of origin, resulting in founder populations that are reproductively isolated due to the contemporary absence of long-distance gene flow.

  6. Determination of staphylococcal enterotoxins in cheese by immunoenzyme assays

    Directory of Open Access Journals (Sweden)

    Janković Vesna

    2012-01-01

    Full Text Available Staphylococcal food poisoning is one of the most common foodborne diseases resulting from the ingestion of staphylococcal enterotoxins (SEs preformed in foods by enterotoxigenic strains of coagulase-positive staphylococci (CPS, mainly Staphylococcus aureus. The presence of enterotoxigenic strains of coagulase-positive staphylococci in raw milk during the production process leads to the contamination of products and outbreaks of alimentary intoxication. The problem of Staphylococcus aureus in cheese remains significant on a global level. Domestic cheese contaminated with enterotoxigenic staphylococci can result in the formation of enterotoxin, which can produce foodborne illness when the product is ingested. Due to microbiological contamination, microbiological criteria are tools that can be used in assessing the safety and quality of foods. In order to avoid foodborne illness, the Serbian Regulation on General and Special Conditions for Food Hygiene (Official Gazette of RS, No. 72/10 provides microbiological criteria for staphylococcal enterotoxins in dairy products.

  7. Staphylococcal Superantigens in Synovial Fluid of 62 Patients With Arthritis

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    A Tabatabaei

    2012-04-01

    Full Text Available Background: Determining the etiologic causes of septic arthritis is of the most importance. Goal of this study was to investigate presence of staphylococcal enterotoxins A, B, C and Toxic Shock Staphylococcal toxin-1 in the synovial fluid of patients with arthritis. Methods: This cross-sectional study was performed in the Pediatric and Orthopedic Wards of Hazrat Rasoul Hospital in Tehran, Iran during 2008- 2010. Gram stains, conventional cultures, direct detection of soluble bacterial antigens were used to detect H. influenza, S. pneumonia, group B streptococci, and N. meningitidis while Latex particle agglutination test was used for staphylococcal supper antigens (by enzyme immunoassays upon synovial fluid tapping of 62 individuals (5 mo to 16 yrs, mean=113.8 yrs. P<0.05 was considered statistically significant. Results: Positive SF cultures (n=11: 5 positive cases of S. aureus; 5 S. pneumonia; 1 H. influenza, and 1 Klebsiella. Positive gram stains: 10%; and positive LPA: 4%. Staphylococcal arthritis was diagnosed in 7 (39% cases upon positive culture or positive gram stain. The most common type was TSST-1 (47% and the least common was enterotoxin B (18%. Isolation of S. aureus (positive culture was correlated to presence of enterotoxin A in synovial fluid but not to enterotoxins B, C or TSST-1. Conclusion: Staph. aureus had a prominent role in arthritis. 47% of cases with negative culture for S. aureus had at least one type of staphylococcal super antigens in the synovial fluid. Searching for antigens of usual organisms or staphylococcal supper antigens could be helpful for diagnosis and subsequent treatment.

  8. Regulatory organization of the staphylococcal sae locus.

    Science.gov (United States)

    Adhikari, Rajan P; Novick, Richard P

    2008-03-01

    This paper describes an investigation of the complex internal regulatory circuitry of the staphylococcal sae locus and the impact of modifying this circuitry on the expression of external genes in the sae regulon. The sae locus contains four genes, the saeR and S two-component signalling module (TCS), and saeP and Q, two upstream genes of hitherto unknown function. It is expressed from two promoters, P(A)sae, which transcribes only the TCS, and P(C)sae, which transcribes the entire locus. A bursa aurealis (bursa) transposon insertion in saeP in a derivative of Staphylococcus aureus NCTC 8325 has a profound effect on sae function. It modifies the activity of the TCS, changing the expression of many genes in the sae regulon, even though transcription of the TCS (from P(A)sae) is not interrupted. Moreover, these effects are not due to disruption of saeP since an in-frame deletion in saeP has essentially no phenotype. The phenotype of S. aureus strain Newman is remarkably similar to that of the saeP : : bursa and this similarity is explained by an amino acid substitution in the Newman saeS gene that is predicted to modify profoundly the signalling function of the protein. This concurrence suggests that the saeP : : bursa insertion affects the signalling function of saeS, a suggestion that is supported by the ability of an saeQR clone, but not an saeR clone, to complement the effects of the saeP : : bursa insertion.

  9. The Management of Staphylococcal Toxic Shock Syndrome. A Case Report

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    Bîrluţiu Victoria

    2016-04-01

    Full Text Available Staphylococcal toxic shock syndrome (TSS is most frequently produced by TSS toxin-1 (TSST-1 and Staphylococcal enterotoxin B (SEB, and only rarely by enterotoxins A, C, D, E, and H. Various clinical pictures can occur depending on severity, patient age and immune status of the host. Severe forms, complicated by sepsis, are associated with a death rate of 50-60%. The case of a Caucasian female infant, aged seven weeks, hospitalized with a diffuse skin rash, characterized as allergodermia, who initially developed TSS with axillary intertrigo, is reported.

  10. Staphylococcal enterotoxin B-specific electrochemiluminescence and lateral flow device assays cross-react with staphylococcal enterotoxin D.

    Science.gov (United States)

    Tallent, Sandra M; Hait, Jennifer; Bennett, Reginald W

    2014-01-01

    Guam school children and faculty members experienced symptoms of vomiting, nausea, abdominal cramps, and diarrhea shortly after eating breakfast prepared by contracted caterers. The first illness was reported within an hour after breakfast, affecting 295 students and two faculty members. Local hospitals treated 130 people, and 61 were admitted for further treatment. Reported symptoms were consistent with staphylococcal food poisoning. Initial food testing using a lateral flow device and electrochemiluminescence method incorrectly implicated staphylococcal enterotoxin B as the causative agent, prompting partial activation of Guam's Emergency Response Center. Traditional ELISAs proved that the food poisoning agent was staphylococcal enterotoxin D. More specific and sensitive assays would have alleviated the issues and confusion that surrounded the reporting and investigation of this outbreak.

  11. Increased hsp70 of glucocorticoid receptor complex induced by scald and heat stress and its possible effect on the affinity of glucocorticoid receptor

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-hui; TANG Hong-tai; LU Jian; XIA Zhao-fan

    2010-01-01

    Background Glucocorticoid (GC) insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor (GR). Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins (hsp) especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro. Methods The affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting.Results Similar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats (30% total body surface area immersion scald) and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols.Conclusions The primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.

  12. Staphylococcal enterotoxins stimulate lymphoma-associated immune dysregulation

    DEFF Research Database (Denmark)

    Krejsgaard, Thorbjørn; Willerslev-Olsen, Andreas; Lindahl, Lise M;

    2014-01-01

    . Here we show that SA isolates from involved skin express staphylococcal enterotoxins (SEs) which induce cross-talk between malignant and benign T cells leading to Stat3-mediated IL-10 production by the malignant T cells. The SEs did not stimulate the malignant T cells directly. Instead, SEs triggered...

  13. Staphylococcal Leukocidin Receptor Identification: Hiding in Plain Sight

    NARCIS (Netherlands)

    Spaan, A.N.

    2015-01-01

    Staphylococcus aureus is a leading cause of bacterial infections in humans worldwide. A key host defense strategy against S. aureus is phagocyte-mediated killing. In return, a major mechanism deployed by S. aureus to target host phagocytes is the production of cytolytic toxins. Staphylococcal leukoc

  14. Staphylococcal Leukocidin Receptor Identification: Hiding in Plain Sight

    NARCIS (Netherlands)

    Spaan, A.N.

    2015-01-01

    Staphylococcus aureus is a leading cause of bacterial infections in humans worldwide. A key host defense strategy against S. aureus is phagocyte-mediated killing. In return, a major mechanism deployed by S. aureus to target host phagocytes is the production of cytolytic toxins. Staphylococcal

  15. Staphylococcal SSL5 inhibits leukocyte activation by chemokines and anaphylatoxins

    NARCIS (Netherlands)

    Bestebroer, Jovanka; van Kessel, Kok P. M.; Azouagh, Hafida; Walenkamp, Annemiek M.; Boer, Ingrid G. J.; Romijn, Roland A.; van Strijp, Jos A. G.; de Haas, Carla J. C.

    2009-01-01

    Staphylococcus aureus secretes several virulence factors modulating immune responses. Staphylococcal superantigen-like (SSL) proteins are a family of 14 exotoxins with homology to superantigens, but with generally unknown function. Recently, we showed that SSL5 binds to P-selectin glycoprotein ligan

  16. Staphylococcal superantigens stimulate immortalized human adipocytes to produce chemokines.

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    Bao G Vu

    Full Text Available BACKGROUND: Human adipocytes may have significant functions in wound healing and the development of diabetes through production of pro-inflammatory cytokines after stimulation by gram-negative bacterial endotoxin. Diabetic foot ulcers are most often associated with staphylococcal infections. Adipocyte responses in the area of the wound may play a role in persistence and pathology. We studied the effect of staphylococcal superantigens (SAgs on immortalized human adipocytes, alone and in the presence of bacterial endotoxin or staphylococcal α-toxin. METHODOLOGY/PRINCIPAL FINDINGS: Primary non-diabetic and diabetic human preadipocytes were immortalized by the reverse transcriptase component of telomerase (TERT and the E6/E7 genes of human papillomavirus. The immortal cells were demonstrated to have properties of non-immortalized pre-adipocytes and could be differentiated into mature and functional adipocytes. Differentiated adipocytes exposed to staphylococcal SAgs produced robust levels of cytokines IL-6 and IL-8, but there were no significant differences in levels between the non-diabetic and diabetic cells. Cytokine production was increased by co-incubation of adipocytes with SAgs and endotoxin together. In contrast, α-toxin alone was cytotoxic at high concentrations, but, at sub-cytotoxic doses, did not stimulate production of IL-6 and IL-8. CONCLUSIONS/SIGNIFICANCE: Endotoxin has been proposed to contribute to diabetes through enhanced insulin resistance after chronic exposure and stimulation of adipocytes to produce cytokines. Our data indicate staphylococcal SAgs TSST-1 and SEB alone and in combination with bacterial endotoxin also stimulate adipocytes to produce cytokines and thus may contribute to the inflammatory response found in chronic diabetic ulcers and in the systemic inflammation that is associated with the development and persistence of diabetes. The immortal human pre-adipocytes reported here will be useful for studies to

  17. Precautions taken by mothers to prevent burn and scald injuries to young children at home: an intervention study.

    Science.gov (United States)

    Carlsson, Anna; Bramhagen, Ann-Cathrine; Jansson, Annkristin; Dykes, Anna-Karin

    2011-07-01

    The aim of this study was to investigate to what extent individual-based extended information given to mothers from city parts of low education can improve precautions taken by them to prevent burn and scald injuries involving young children in the home and further to compare the results with a group of mothers who had not received extended information. This intervention study, with a comparison group, has a quasi-experimental design. Individual-based information, with an empowerment approach, was given to a group of mothers living in two separate areas of a city in southern Sweden with a low level of education. In total, 99 mothers of children under the age of 7 months participated. The mothers were selected through the local child healthcare authorities. Observations were made and bivariate analyses were established. The results showed that the intervention had a significant impact on improving the precautions the participating mothers introduced to protect their children against burn and scald injuries in the home and further, in relation to a comparison group. Through empowerment, workshops, and home visits aimed to increase their consciousness and knowledge, the participating mothers' precautions taken against child injuries in the home improved. It is of great importance that a framework for considering the problem of burn and scald injuries to children is presented from a preventive perspective which, in combination with evidence-based interventions, may enable the creation of injury prevention programmes for implementation by the community health care.

  18. Scald burns in young children--a review of Arizona burn center pediatric patients and a proposal for prevention in the Hispanic community.

    Science.gov (United States)

    Rimmer, Ruth B; Weigand, Shannon; Foster, Kevin N; Wadsworth, Michelle M; Jacober, Katie; Matthews, Marc R; Drachman, David; Caruso, Daniel M

    2008-01-01

    Arizona Burn Center staff observed an increasing number of pediatric scald burn admissions. Therefore, a retrospective study was conducted to identify scald demographics and etiologies. Focus groups were subsequently conducted to determine burn prevention knowledge in the target community. Arizona Burn Center scald admission variables (ages 0-5 years) reviewed included age, sex, ethnicity, TBSA, body parts burned, occurrence month and location, caregiver present, child and caregiver activities when injured, payor source, length of stay, parental language, and zip code. High-risk zip code area focus groups were then conducted. There were a total of 170 pediatric patients, aged 0 to 5 years, admitted to the burn center during 2005 to 2006. Of this total, 124 of the patients were admitted for scald burns, accounting for 59% of all pediatric burn admissions. Scald burn patient's demographics included male (52%), female (48%) with a mean age of 1.7 years. Majority ethnicity was Hispanic (63%). The mean TBSA was 8% with mean length of stay of 8 days. The remaining pediatric admissions for children aged 0 to 5 were for burns caused by fire or flame 15%, contact with a hot object 13%, friction burns 7%, chemical burns 3%, and electrical burns 3%. Demographics for the combined etiologies included an identical sex breakdown with male (52%) and female (48%). The majority ethnicity in the nonscald group was also Hispanic at 59% with a mean length of stay of 7 days and an average TBSA of 9.5%. Main etiologies of scald burns included hot water (25%), soup (24%), and coffee or tea (21%). Most common child behaviors were pulling hot substance from stove (24%), from countertop (13%), and having liquid spilled on them (13%) typically while caregiver was cooking. Mean TBSA was 8% with mean length of stay (8 days). Scalds occurred in the kitchen (83%) and mainly in child's home (94%). Mother was primary caregiver (78%). Payor source was Medicaid (86%) and the average admission cost

  19. [Rat immunoreactivity in experimental modeling of acute autohemorrhagic bihemispheric stroke in the presence of staphylococcal infection].

    Science.gov (United States)

    Makarenko, A N; Mironiuk, Iu N; Molozhavaia, O S; Pozur, V K; Iablonskaia, S V

    2008-01-01

    Experimental studies were made of rat immunoreactivity in experimental modeling of acute hemorrhagic stroke in staphylococcal infection. Bactericidal activity and reserve bactericidal activity of neutrophils in NBT-test enhance while mid-molecular CIC level decreases because of phagocyte activation. ABA high titers in blood reflect activation of autoimmune processes. Splenic mass increased suggesting activation of proliferation of splenic lymphoid cells or enhanced migration of immature lymphoid cells from bone marrow to the spleen. Experimental animals had suppressed functional activity of main lymphoid cells populations in blast-transformation reaction which may affect the course of early recovery, promote activation of infectious bacterial process, autoaggression. The results of the studies can serve the basis for further investigations of immune mechanisms involved in development of hemorrhagic stroke in the presence of infectious process.

  20. Evaluation of Handheld Assays for the Detection of Ricin and Staphylococcal Enterotoxin B in Disinfected Waters

    Science.gov (United States)

    2011-01-01

    Evaluation of Handheld Assays for the Detection of Ricin and Staphylococcal Enterotoxin B in DisinfectedWaters MaryMargaretWade,1 Tracey D. Biggs,1...Staphylococcal Enterotoxin B (SEB) in water. Performance of HHAs was evaluated in formulated tap water with and without chlorine, reverse osmosis water (RO) with...Handheld Assays For The Detection of Ricin And Staphylococcal Enterotoxin B In Disinfected Waters 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM

  1. Activation of MyD88 Signaling upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules

    Science.gov (United States)

    2011-01-20

    Activation of MyD88 Signaling upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules Teri L. Kissner, Gordon Ruthel, Shahabuddin Alam...mediated signaling, which activates pro-inflammatory cytokine responses. Recently we reported that staphylococcal enterotoxins (SEA or SEB), which...upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules. PLoS ONE 6(1): e15985. doi:10.1371/journal.pone.0015985 Editor: Jacques Zimmer

  2. Bacteriophage-Derived Peptidase CHAPK Eliminates and Prevents Staphylococcal Biofilms

    Directory of Open Access Journals (Sweden)

    Mark Fenton

    2013-01-01

    Full Text Available New antibacterial agents are urgently needed for the elimination of biofilm-forming bacteria that are highly resistant to traditional antimicrobial agents. Proliferation of such bacteria can lead to significant economic losses in the agri-food sector. This study demonstrates the potential of the bacteriophage-derived peptidase, CHAPK, as a biocidal agent for the rapid disruption of biofilm-forming staphylococci, commonly associated with bovine mastitis. Purified CHAPK applied to biofilms of Staphylococcus aureus DPC5246 completely eliminated the staphylococcal biofilms within 4 h. In addition, CHAPK was able to prevent biofilm formation by this strain. The CHAPK lysin also reduced S. aureus in a skin decolonization model. Our data demonstrates the potential of CHAPK as a biocidal agent for prevention and treatment of biofilm-associated staphylococcal infections or as a decontaminating agent in the food and healthcare sectors.

  3. Monkey Feeding Assay for Testing Emetic Activity of Staphylococcal Enterotoxin.

    Science.gov (United States)

    Seo, Keun Seok

    2016-01-01

    Staphylococcal enterotoxins (SEs) are unique bacterial toxins that cause gastrointestinal toxicity as well as superantigenic activity. Since systemic administration of SEs induces superantigenic activity leading to toxic shock syndrome that may mimic enterotoxic activity of SEs such as vomiting and diarrhea, oral administration of SEs in the monkey feeding assay is considered as a standard method to evaluate emetic activity of SEs. This chapter summarizes and discusses practical considerations of the monkey feeding assay used in studies characterizing classical and newly identified SEs.

  4. FDA approved Immunosuppressants Targeting Staphylococcal Superantigens: Mechanisms and Insights

    Science.gov (United States)

    2016-12-02

    for public release; distribution is unlimited. UNCLASSIFIED 91 . Solinas G, Karin M. JNKl and IKK.beta: molecular links between obesity and metabolic ...is unlimited. UNCLASSIFIED Introduction Staphylococcal endotoxin B (SEB) and the distantly related toxic shock syndrome toxin 1 (TSST-1) are common...etiological agents that cause toxic shock syndrome [1, 2]. The disease is characterized by fever, hypotension, desquamation of skin, and multiple

  5. Sequence Variability in Staphylococcal Enterotoxin Genes seb, sec, and sed

    Directory of Open Access Journals (Sweden)

    Sophia Johler

    2016-06-01

    Full Text Available Ingestion of staphylococcal enterotoxins preformed by Staphylococcus aureus in food leads to staphylococcal food poisoning, the most prevalent foodborne intoxication worldwide. There are five major staphylococcal enterotoxins: SEA, SEB, SEC, SED, and SEE. While variants of these toxins have been described and were linked to specific hosts or levels or enterotoxin production, data on sequence variation is still limited. In this study, we aim to extend the knowledge on promoter and gene variants of the major enterotoxins SEB, SEC, and SED. To this end, we determined seb, sec, and sed promoter and gene sequences of a well-characterized set of enterotoxigenic Staphylococcus aureus strains originating from foodborne outbreaks, human infections, human nasal colonization, rabbits, and cattle. New nucleotide sequence variants were detected for all three enterotoxins and a novel amino acid sequence variant of SED was detected in a strain associated with human nasal colonization. While the seb promoter and gene sequences exhibited a high degree of variability, the sec and sed promoter and gene were more conserved. Interestingly, a truncated variant of sed was detected in all tested sed harboring rabbit strains. The generated data represents a further step towards improved understanding of strain-specific differences in enterotoxin expression and host-specific variation in enterotoxin sequences.

  6. Staphylococcal septicaemia complicated with purulent pericarditis in ...

    African Journals Online (AJOL)

    2016-08-18

    Aug 18, 2016 ... diac diagnostic equipment in developing countries like. Nigeria.2,4,5 There ... This study presents a one year old girl who was initially managed as a case .... due to the pharmacokinetics of antibiotics with a more prolonged course ... culture was negative due to early onset of action of anti- biotics, while the ...

  7. Target metabolite and gene transcription profiling during the development of superficial scald in apple (Malus x domestica Borkh).

    Science.gov (United States)

    Busatto, Nicola; Farneti, Brian; Tadiello, Alice; Vrhovsek, Urska; Cappellin, Luca; Biasioli, Franco; Velasco, Riccardo; Costa, Guglielmo; Costa, Fabrizio

    2014-07-20

    Fruit quality features resulting from ripening processes need to be preserved throughout storage for economical reasons. However, during this period several physiological disorders can occur, of which superficial scald is one of the most important, due to the development of large brown areas on the fruit skin surface. This study examined the variation in polyphenolic content with the progress of superficial scald in apple, also with respect to 1-MCP, an ethylene competitor interacting with the hormone receptors and known to interfere with this etiology. The change in the accumulation of these metabolites was further correlated with the gene set involved in this pathway, together with two specific VOCs (Volatile Organic Compounds), α-farnesene and its oxidative form, 6-methyl-5-hepten-2-one. Metabolite profiling and qRT-PCR assay showed these volatiles are more heavily involved in the signalling system, while the browning coloration would seem to be due more to a specific accumulation of chlorogenic acid (as a consequence of the activation of MdPAL and MdC3H), and its further oxidation carried out by a polyphenol oxidase gene (MdPPO). In this physiological scenario, new evidence regarding the involvement of an anti-apoptotic regulatory mechanism for the compartmentation of this phenomenon in the skin alone was also hypothesized, as suggested by the expression profile of the MdDAD1, MdDND1 and MdLSD1 genes. The results presented in this work represent a step forward in understanding the physiological mechanisms of superficial scald in apple, shedding light on the regulation of the specific physiological cascade.

  8. At-a-glance, Emergency department surveillance of thermal burns and scalds, electronic Canadian Hospitals Injury Reporting and Prevention Program, 2013.

    Science.gov (United States)

    Crain, J; McFaull, S; Rao, D P; Do, M T; Thompson, W

    2017-01-01

    Although fatality and hospitalization rates for burns in Canada have declined over time, less serious cases still commonly present to the emergency department (ED). The Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP) is an injury and poisoning surveillance system administered by the Public Health Agency of Canada, operating in emergency departments of 17 hospitals. Overall, cases reported in 2013 were scalds and contact burns from hot objects. The leading direct causes of scalds were hot beverages and hot water. The leading causes of contact burns were stoves/ovens and fireplaces/accessories. While the overall proportion of burns was highest among females, males comprised a higher proportion of burns from all mechanisms except scalds.

  9. Surface plasmon resonance (SPR) detection of Staphylococcal Enterotoxin A in food samples

    Science.gov (United States)

    An automated and rapid method for detection of staphylococcal enterotoxins (SE) is needed. A sandwich assay was developed using a surface plasmon resonance (SPR) biosensor for detection of staphylococcal enterotoxin A (SEA) at subpicomolar concentration. Assay conditions were optimized for capturing...

  10. Lactobacilli require physical contact to reduce staphylococcal TSST-1 secretion and vaginal epithelial inflammatory response

    NARCIS (Netherlands)

    Younes, Jessica A.; Reid, Gregor; van der Mei, Henny C.; Busscher, Henk J.

    2016-01-01

    Staphylococcus aureus biofilms can be found on vaginal epithelia, secreting toxins and causing inflammation. The co-vaginal species Lactobacillus can alter staphylococcal-induced epithelial secretion of inflammatory cytokines and quench staphylococcal toxic shock syndrome toxin-1 secretion. It is hy

  11. A rare case of staphylococcal cold abscess of anterior chest wall in an immunocompetent adult

    Directory of Open Access Journals (Sweden)

    Sibes Kumar Das

    2012-01-01

    Full Text Available Staphylococcus aureus is an important causative organism for skin and soft tissue infection, which presents with the classical local signs of acute inflammation. Staphylococcal abscess without signs of inflammation (staphylococcal cold abscess is a very rare entity, sometimes seen in immunocompromised host. Here, we report a case of a 50-year-old male patient who presented with bilateral asymptomatic cold abscess of staphylococcal origin over the anterior chest wall. The patient had no immunodeficiency and there was no distant/underlying source of staphylococcal infection. Smear and culture of pus proved the staphylococcal etiology and excluded tuberculosis, its close differential diagnosis. The patient was treated with antistaphylococcal antibiotics and local drainage of pus with complete recovery.

  12. Host responses associated with chronic staphylococcal mastitis in rabbits.

    Science.gov (United States)

    Guerrero, Irene; Ferrian, Selena; Penadés, Mariola; García-Quirós, Ana; Pascual, Juan J; Selva, Laura; Viana, David; Corpa, Juan M

    2015-06-01

    Staphylococcal infection causes substantial economic losses in commercial rabbit production systems, and is associated with a wide variety of lesions, including chronic suppurative mastitis, which mainly affects breeding females. Most chronic staphylococcal infections in rabbits are caused by the ST121 lineage of Staphylococcus aureus, although other less common lineages, such as ST96 can also be involved. The aims of the present study were to characterise the host immune response in natural cases of mastitis in rabbits caused by S. aureus, to evaluate any relationship between peripheral and local immunity and to investigate the effect of different S. aureus genotypes on these immune responses. Adult multiparous female rabbits that were affected with chronic staphylococcal mastitis (n = 204) were enrolled into the study. Histological and immunohistochemical evaluations of mammary glands were undertaken, as well as flow cytometric analyses of blood. S. aureus isolates from the mammary glands were identified by multilocus sequence typing. Differences in the number of infiltrating cells were detected, depending on the type of pathology, with more immature lesions demonstrating greater cellularity, characterised by greater numbers of T lymphocytes, macrophages and plasma cells. A relationship was seen between the cells in blood and mammary tissues, the most notable being the positive correlation between monocytes and tissue macrophages. When glands were infected with ST96 strains, fewer granulocytes (P < 0.01) and greater numbers of B cells (P < 0.01), T cells (P < 0.001), CD4(+) T cells (P < 0.001) and CD8(+) T cells (P < 0.01) were detected, compared with mammary glands that were infected by ST121 strains of S. aureus. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Effects of staphylococcal enterotoxin B on rodent mast cells.

    OpenAIRE

    Komisar, J; Rivera, J.; Vega, A.; Tseng, J

    1992-01-01

    Staphylococcal enterotoxin B (SEB) was tested in rodent mast cell cultures for the release of serotonin. Both rat RBL-2H3 mast cells and murine peritoneal cells released serotonin after SEB stimulation in culture. Release of serotonin in RBL-2H3 cells depended on the concentration of SEB; an appreciable release was seen at 50 micrograms/ml. The release of serotonin was not due to cell death. Serotonin release could be enhanced by bradykinin but not by vasoactive intestinal peptide, substance ...

  14. Real time-based targeted Metabolomic approach as risk assessment tools for controlling superficial scald in ‘Granny Smith’ apples during storage

    Science.gov (United States)

    The risk of superficial scald incidence in ‘Granny Smith’ apple [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] fruit during cold or controlled atmosphere (CA) storage increases with increased storage duration and oxygen level during CA storage. The objective of this study was to invest...

  15. Staphylococcal endogenous endophthalmitis in association with pyogenic vertebral osteomyelitis.

    Science.gov (United States)

    Steeples, L R; Jones, N P

    2016-01-01

    PURPOSE To describe pyogenic vertebral osteomyelitis as a rare infection associated with endogenous endophthalmitis.METHODS A retrospective review of three patients with endogenous endophthalmitis and sepsis due to underlying Staphylococcal vertebral osteomyelitis presenting during a 21-month time period. The ophthalmic and systemic features and management and outcomes are presented.RESULTS One patient developed unilateral endophthalmitis with cervical spine osteomyelitis, Staphylococcus aureus being isolated from blood cultures. The second presented with bilateral endophthalmitis with disseminated Methicillin-resistant S. aureus (MRSA) infection, with thoracic and lumbar discitis and para-spinal abscesses. MRSA was cultured from vitreous, blood, and synovial fluid. Both patients received prolonged courses of intravenous antibiotics. Intravitreal antibiotic therapy was used in the second patient. Excellent visual and systemic outcomes were achieved in both cases with no ocular complications. The third patient developed lumbar osteomyelitis following spinal surgery and presented with disseminated S. aureus sepsis including unilateral endogenous endophthalmitis. Despite systemic antibiotics and intensive care the patient died.CONCLUSIONS Endogenous endophthalmitis should be suspected in septic patients developing eye symptoms. Endogenous endophthalmitis with staphylococcal bone infection is a rare but serious condition. Osteomyelitis should be considered as an infective source in any such patient reporting bone pain or reduced spinal mobility. Prompt investigation and treatment can achieve favourable visual and systemic outcomes.

  16. Tool for quantification of staphylococcal enterotoxin gene expression in cheese.

    Science.gov (United States)

    Duquenne, Manon; Fleurot, Isabelle; Aigle, Marina; Darrigo, Claire; Borezée-Durant, Elise; Derzelle, Sylviane; Bouix, Marielle; Deperrois-Lafarge, Véronique; Delacroix-Buchet, Agnès

    2010-03-01

    Cheese is a complex and dynamic microbial ecosystem characterized by the presence of a large variety of bacteria, yeasts, and molds. Some microorganisms, including species of lactobacilli or lactococci, are known to contribute to the organoleptic quality of cheeses, whereas the presence of other microorganisms may lead to spoilage or constitute a health risk. Staphylococcus aureus is recognized worldwide as an important food-borne pathogen, owing to the production of enterotoxins in food matrices. In order to study enterotoxin gene expression during cheese manufacture, we developed an efficient procedure to recover total RNA from cheese and applied a robust strategy to study gene expression by reverse transcription-quantitative PCR (RT-qPCR). This method yielded pure preparations of undegraded RNA suitable for RT-qPCR. To normalize RT-qPCR data, expression of 10 potential reference genes was investigated during S. aureus growth in milk and in cheese. The three most stably expressed reference genes during cheese manufacture were ftsZ, pta, and gyrB, and these were used as internal controls for RT-qPCR of the genes sea and sed, encoding staphylococcal enterotoxins A and D, respectively. Expression of these staphylococcal enterotoxin genes was monitored during the first 72 h of the cheese-making process, and mRNA data were correlated with enterotoxin production.

  17. Further Evidence for Staphylococcal Food Poisoning Outbreaks Caused by egc-Encoded Enterotoxins

    Directory of Open Access Journals (Sweden)

    Sophia Johler

    2015-03-01

    Full Text Available Staphylococcal food poisoning represents the most prevalent foodborne intoxication worldwide. It is caused by oral intake of enterotoxins preformed by Staphylococcus aureus in food. The relevance of newly described enterotoxins in outbreaks of staphylococcal food poisoning is controversially discussed. Although the staphylococcal enterotoxins SEG, SEI, SEM, SEN, and SEO elicit emesis in a monkey feeding assay, there has been no conclusive proof of their emetic activity in humans. In this study, we provide further evidence suggesting that one of these enterotoxins or a combination of SEG, SEI, SEM, SEN, and SEO cause staphylococcal food poisoning. We investigated two outbreaks registered with the Swiss Federal Office of Public Health, in which only Staphylococcus aureus strains harboring the egc cluster, including seg, sei, sem, sen, and seo linked to typical signs of staphylococcal food poisoning were isolated. The outbreaks were caused by consumption of raw goat cheese and semi-hard goat cheese, and were linked to strains assigned to CC45 (agr type I and CC9 (agr type II, respectively. These outbreaks provide further evidence that newly-described staphylococcal enterotoxins are likely to cause staphylococcal food poisoning in humans.

  18. Further evidence for staphylococcal food poisoning outbreaks caused by egc-encoded enterotoxins.

    Science.gov (United States)

    Johler, Sophia; Giannini, Petra; Jermini, Marco; Hummerjohann, Jörg; Baumgartner, Andreas; Stephan, Roger

    2015-03-20

    Staphylococcal food poisoning represents the most prevalent foodborne intoxication worldwide. It is caused by oral intake of enterotoxins preformed by Staphylococcus aureus in food. The relevance of newly described enterotoxins in outbreaks of staphylococcal food poisoning is controversially discussed. Although the staphylococcal enterotoxins SEG, SEI, SEM, SEN, and SEO elicit emesis in a monkey feeding assay, there has been no conclusive proof of their emetic activity in humans. In this study, we provide further evidence suggesting that one of these enterotoxins or a combination of SEG, SEI, SEM, SEN, and SEO cause staphylococcal food poisoning. We investigated two outbreaks registered with the Swiss Federal Office of Public Health, in which only Staphylococcus aureus strains harboring the egc cluster, including seg, sei, sem, sen, and seo linked to typical signs of staphylococcal food poisoning were isolated. The outbreaks were caused by consumption of raw goat cheese and semi-hard goat cheese, and were linked to strains assigned to CC45 (agr type I) and CC9 (agr type II), respectively. These outbreaks provide further evidence that newly-described staphylococcal enterotoxins are likely to cause staphylococcal food poisoning in humans.

  19. New biological temporary skin cover Xe-Derma(®) in the treatment of superficial scald burns in children.

    Science.gov (United States)

    Zajicek, Robert; Matouskova, Eva; Broz, Ludomir; Kubok, Richard; Waldauf, Petr; Königova, Radana

    2011-03-01

    Xe-Derma® is a new dry sterile biological cover derived from acellular pig dermis. Hydrated Xe-Derma® displays bio-mechanical features similar to the normal skin. The aim of the present study was to compare the efficacy of Xe-Derma® with hydrocolloid dressing Askina THINSite® for treatment of superficial burns in children in a prospective study. In a prospective study, 86 patients (5 months to 7 years of age) with superficial scald burns on a surface area of 1-35% BSA were enrolled. In the course of the study, 43 patients were treated with Xe-Derma® and 43 patients with Askina THINSite®. We collected data including the percentage of BSA covered with biological or synthetic material, epithelization time, the number of complete conversions (deepening of 100% of covered area into deep dermal wound) under each cover, the number and extent of partial conversions (deepening of less then 100% of covered area into deep dermal wound), infectious complications, the number of reapplications of the temporary cover and the extent in square centimetres of dressing material needed for successful healing of 1% BSA. No significant difference in the epithelization time, percentage of conversion from superficial to deep dermal burns and percentage of infectious complication was detected between the two groups. However, patients in the Xe-Derma® group were burned on a more extensive burn surface area (p ≤ 0.028). Xe-Derma® showed adherence to the wound and therefore there has been no need to be changed The number of reapplications and therefore also the number of square centimetres needed for successful healing of 1% BSA were statistically higher in the Askina THINSite® group (p skin covers in the treatment of superficial scald burns in children. Copyright © 2010 Elsevier Ltd and ISBI. All rights reserved.

  20. Staphylococcal disease in Africa: another neglected 'tropical' disease.

    Science.gov (United States)

    Herrmann, Mathias; Abdullah, Salim; Alabi, Abraham; Alonso, Pedro; Friedrich, Alexander W; Fuhr, Günther; Germann, Anja; Kern, Winfried V; Kremsner, Peter G; Mandomando, Inacio; Mellmann, Alexander C; Pluschke, Gerd; Rieg, Siegbert; Ruffing, Ulla; Schaumburg, Frieder; Tanner, Marcel; Peters, Georg; von Briesen, Hagen; von Eiff, Christof; von Müller, Lutz; Grobusch, Martin P

    2013-01-01

    The term 'neglected tropical diseases' predominantly refers to single-entity, mostly parasitic diseases. However, a considerable morbidity and mortality burden is carried by patients infected with Gram-positive cocci and Gram-negative bacilli that are prevalent all over the world, yet have impact in tropical and developing countries, particularly in children, with much higher incidence rates than those reported from developed countries. Staphylococcus aureus is among these pathogens. The African-German StaphNet consortium uses microbiological characterization of African S. aureus isolates, including identification of virulence factors, alongside the gathering of epidemiological and clinical data in an innovative research network between a European country (Germany) and several African partners. By creating an accessible strain repository and by implementing personnel training and capacity building, this network aims to put staphylococcal disease on the international agenda as a truly neglected condition with a major global impact on public health.

  1. A Multiplex Assay for Detection of Staphylococcal and Streptococcal Exotoxins.

    Directory of Open Access Journals (Sweden)

    Preeti Sharma

    Full Text Available Staphylococcal and streptococcal exotoxins, also known as superantigens, mediate a range of diseases including toxic shock syndrome, and they exacerbate skin, pulmonary and systemic infections caused by these organisms. When present in food sources they can cause enteric effects commonly known as food poisoning. A rapid, sensitive assay for the toxins would enable testing of clinical samples and improve surveillance of food sources. Here we developed a bead-based, two-color flow cytometry assay using single protein domains of the beta chain of T cell receptors engineered for high-affinity for staphylococcal (SEA, SEB and TSST-1 and streptococcal (SpeA and SpeC toxins. Site-directed biotinylated forms of these high-affinity agents were used together with commercial, polyclonal, anti-toxin reagents to enable specific and sensitive detection with SD50 values of 400 pg/ml (SEA, 3 pg/ml (SEB, 25 pg/ml (TSST-1, 6 ng/ml (SpeA, and 100 pg/ml (SpeC. These sensitivities were in the range of 4- to 80-fold higher than achieved with standard ELISAs using the same reagents. A multiplex format of the assay showed reduced sensitivity due to higher noise associated with the use of multiple polyclonal agents, but the sensitivities were still well within the range necessary for detection in food sources or for rapid detection of toxins in culture supernatants. For example, the assay specifically detected toxins in supernatants derived from cultures of Staphylococcus aureus. Thus, these reagents can be used for simultaneous detection of the toxins in food sources or culture supernatants of potential pathogenic strains of Staphylococcus aureus and Streptococcus pyogenes.

  2. Staphylococcus aureus and Staphylococcal Food-Borne Disease: An Ongoing Challenge in Public Health

    OpenAIRE

    2014-01-01

    Staphylococcal food-borne disease (SFD) is one of the most common food-borne diseases worldwide resulting from the contamination of food by preformed S. aureus enterotoxins. It is one of the most common causes of reported food-borne diseases in the United States. Although several Staphylococcal enterotoxins (SEs) have been identified, SEA, a highly heat-stable SE, is the most common cause of SFD worldwide. Outbreak investigations have found that improper food handling practices in the retail ...

  3. Early

    Directory of Open Access Journals (Sweden)

    Kamel Abd Elaziz Mohamed

    2014-04-01

    Conclusion: Early PDT is recommended for patients who require prolonged tracheal intubation in the ICU as outcomes like the duration of mechanical ventilation length of ICU stay and hospital stay were significantly shorter in early tracheostomy.

  4. Melaleuca alternifolia essential oil possesses potent anti-staphylococcal activity extended to strains resistant to antibiotics.

    Science.gov (United States)

    Ferrini, A M; Mannoni, V; Aureli, P; Salvatore, G; Piccirilli, E; Ceddia, T; Pontieri, E; Sessa, R; Oliva, B

    2006-01-01

    Melaleuca alternifolia Cheel essential oil (TTO) and its major component terpinen-4-ol were examined against a large number of clinical isolates of Staphylococcus aureus to establish their anti-staphylococcal activities. Classic and established procedures were used to study M.I.C., time-kill curves, synergism and mutational frequency. The anti-staphylococcal activity of terpinen-4-ol and TTO were superior to those of antibiotics belonging to the major families (all the tested drugs are for topical use or included in ointments, eye drops or used during surgery); terpinen 4-ol and TTO were active against strains resistant to mupirocin, fusidic acid, vancomycin, methicillin and linezolid. TTO and terpinen-4-ol were bactericidal as revealed by time-kill curves; the frequency of mutational frequency to TTO was < 2.9 x 10 9. The study demonstrates good anti-staphylococcal activity of TTO and terpinen-4-ol against a large number of S.aureus isolates and suggests the possible application of these agents for topical treatment of staphylococcal infections. This is the first extensive study on the anti-staphylococcal activity of TTO. The results suggest that this compound may have application as a topical agent for the control of superficial staphylococcal infections, including activity against organisms resistant to antibiotics which can be used, or are specific, for topical use.

  5. Functional piglet model for the clinical syndrome and postmortem findings induced by staphylococcal enterotoxin B.

    Science.gov (United States)

    van Gessel, Yvonne A; Mani, Sachin; Bi, Shuguang; Hammamieh, Rasha; Shupp, Jeffrey W; Das, Rina; Coleman, Gary D; Jett, Marti

    2004-11-01

    Staphylococcal enterotoxin (SE) B causes serious gastrointestinal illness, and intoxication with this exotoxin can lead to lethal toxic shock syndrome. In order to overcome significant shortcomings of current rodent and nonhuman primate models, we developed a piglet model of lethal SEB intoxication. Fourteen-day-old Yorkshire piglets were given intravenous SEB, observed clinically, and sacrificed at 4, 6, 24, 48, 72, or 96 hrs posttreatment. Clinical signs were biphasic with pyrexia, vomiting, and diarrhea within 4 hrs, followed by terminal hypotension and shock by 96 hrs. Mild lymphoid lesions were identified as early as 24 hrs, with severe lymphadenopathy, splenomegaly, and prominent Peyer's patches found by 72 hrs. Widespread edema-most prominent in the mesentery, between loops of spiral colon, and in retroperitoneal connective tissue-was found in animals at 72 hrs. Additional histologic changes included perivascular aggregates of large lymphocytes variably present in the lung and brain, circulating lymphoblasts, and lymphocytic portal hepatitis. Preliminary molecular investigation using gene array has uncovered several gene profile changes that may have implications in the pathophysiology leading to irreversible shock. Five genes were selected for further study, and all showed increased mRNA levels subsequent to SEB exposure. The use of this piglet model will continue to elucidate the pathogenesis of SEB intoxication and facilitate the testing of new therapeutic regimens that may better correlate with human lesions.

  6. Exfoliative toxins of Staphylococcus aureus and their mechanism of action in staphylococcal scalded skin syndrome%葡萄球菌性烫伤样皮肤综合征的致病毒素及作用机制

    Institute of Scientific and Technical Information of China (English)

    苏怡帆; 王华

    2007-01-01

    由金黄色葡萄球菌分泌的表皮剥脱毒素是引起葡萄球菌性烫伤样皮肤综合征患者皮肤剥脱的直接原因.该病的临床表现型与毒素血清分型相关.表皮剥脱毒素作为一种丝氨酸蛋白酶,能作用于表皮颗粒层,特异性结合并水解桥粒芯糖蛋白1,从桥粒芯糖蛋白1的胞外区EC3和EC4之间劈开谷氨酸381-X肽键,从而形成表皮内大疱.此外,表皮剥脱毒素还可能作为超抗原参与发病.

  7. Protective role of ET antibody against staphylococcal scalded skin syndrome%ET抗体对葡萄球菌性烫伤样皮肤综合征的免疫保护作用

    Institute of Scientific and Technical Information of China (English)

    肖异珠; 王毅; 杨致邦; 欧阳莹; 李咏梅; 罗晓燕; 谭琦; 向娟

    2014-01-01

    目的 探讨ET抗体对葡萄球菌性烫伤样皮肤综合征的免疫保护作用.方法 用重组ETA和ETB建立检测血清ETA和ETB抗体的间接ELISA法,用建立的ELISA法测定商业化人高效价丙种球蛋白(IVIg)、葡萄球菌性烫伤样皮肤综合征(SSSS)患儿、特应性皮炎(AD)患儿和健康儿童血清中ETA和ETB抗体的效价.用重组ETA和ETB皮下注射新生小鼠建立SSSS动物模型,在注射重组ETA和ETB的同时和3h后在小鼠腹腔内注射ETA和ETB抗体,观察小鼠发病情况和皮肤组织病理改变.结果 IVIg中测出高效价的ETA和ETB抗体;SSSS患儿、AD患儿和健康儿童血清中ETB抗体效价(A450nm)分别为0.863±0.276、1.027±0.222、0.990±0.151.SSSS患儿血清中ETB抗体效价明显低于AD患儿和健康儿童,差异有统计学意义(P<0.05),但三者间ETA抗体的效价差异无统计学意义(P>0.05).注射ETA和ETB抗体的新生小鼠发病、皮肤组织病理改变和死亡率均低于仅注射重组ETA和ETB小鼠.结论 ET抗体对葡萄球菌性烫伤样皮肤综合征具有免疫保护作用,以ETB抗体为主,可减轻皮肤损伤,降低死亡率.

  8. Identifying exposure targets for treatment of staphylococcal pneumonia with ceftobiprole.

    Science.gov (United States)

    Rodvold, Keith A; Nicolau, David P; Lodise, Thomas P; Khashab, Mohammed; Noel, Gary J; Kahn, James B; Gotfried, Mark; Murray, Sara A; Nicholson, Susan; Laohavaleeson, Somvadee; Tessier, Pamela R; Drusano, G L

    2009-08-01

    Ceftobiprole is a cephalosporin with potent activity against methicillin (meticillin)-resistant Staphylococcus aureus (MRSA). In order to treat patients with severe staphylococcal pneumonia, it is important to understand the drug exposure required to mediate the killing of multiple log(10) cells in a preclinical-infection model. We measured drug exposure in terms of the percentage of penetration of the drug into epithelial lining fluid (ELF) and in terms of the time for which the drug concentration was above the MIC (time>MIC) in plasma and ELF. In a murine model of staphylococcal pneumonia, we demonstrated that ceftobiprole penetrated into ELF from the plasma at a median level of nearly 69% (25th to 75th percentile range, 25 to 187%), as indexed to the ratio of values for the area under the concentration-time curve in ELF and plasma. The total-drug times>MIC in ELF that were required to kill 1 log(10) and 2 log(10) CFU/g of lung tissue were 15% and 25% of the dosing interval. We also examined the penetration of ELF by ceftobiprole in volunteers, demonstrating mean and median penetration percentages of 25.5% and 15.3%, respectively (25th to 75th percentile range, 8 to 30%). Attainment rates were calculated for kill targets of 1 log(10) and 2 log(10) CFU/g, taken from the murine model, but using the volunteer ceftobiprole ELF penetration data. The standard dose for ceftobiprole is 0.5 g every 8 h as a 2-h infusion. The attainment rates remained above 90% for 1-log(10) and 2-log(10) CFU/g kill targets at MICs of 1 and 0.5 mg/liter, respectively. Taking the expectation over the distribution of ceftobiprole MICs for 4,958 MRSA isolates showed an overall target attainment of 85.6% for a 1-log(10) CFU/g kill and 79.7% for a 2-log(10) CFU/g kill. It is important to derive exposure targets in preclinical-infection models of the infection site so that these targets can be explored in clinical trials in order to optimize the probability of a good clinical outcome.

  9. Puffy Hand Syndrome Revealed by a Severe Staphylococcal Skin Infection

    Directory of Open Access Journals (Sweden)

    Reyhan Amode

    2013-01-01

    Full Text Available Puffy hand syndrome develops after long-term intravenous drug addiction. It is characterized by a nonpitting edema, affecting the dorsal side of fingers and hands with puffy aspect. Frequency and severity of the complications of this syndrome are rarely reported. Local infectious complications such as cellulitis can be severe and can enable the diagnosis. Herein, we report the case of a 41-year-old man who went to the emergency department for abdominal pain, fever, and bullous lesions of legs and arms with edema. Bacteriologic examination of a closed bullous lesion evidenced a methicillin sensitive Staphylococcus aureus. The abdomen computed tomography excluded deep infections and peritoneal effusion. The patient was successfully treated by intravenous oxacillin and clindamycin. He had a previous history of intravenous heroin addiction. We retained the diagnosis of puffy hand syndrome revealed by a severe staphylococcal infection with toxic involvement mimicking a four limbs cellulitis. Puffy hand syndrome, apart from the chronic lymphedema treatment, has no specific medication available. Prophylactic measures against skin infections are essential.

  10. Two common structural motifs for TCR recognition by staphylococcal enterotoxins

    Science.gov (United States)

    Rödström, Karin E. J.; Regenthal, Paulina; Bahl, Christopher; Ford, Alex; Baker, David; Lindkvist-Petersson, Karin

    2016-01-01

    Superantigens are toxins produced by Staphylococcus aureus, called staphylococcal enterotoxins (abbreviated SEA to SEU). They can cross-link the T cell receptor (TCR) and major histocompatibility complex class II, triggering a massive T cell activation and hence disease. Due to high stability and toxicity, superantigens are potential agents of bioterrorism. Hence, antagonists may not only be useful in the treatment of disease but also serve as countermeasures to biological warfare. Of particular interest are inhibitors against SEA and SEB. SEA is the main cause of food poisoning, while SEB is a common toxin manufactured as a biological weapon. Here, we present the crystal structures of SEA in complex with TCR and SEE in complex with the same TCR, complemented with computational alanine-scanning mutagenesis of SEA, SEB, SEC3, SEE, and SEH. We have identified two common areas that contribute to the general TCR binding for these superantigens. This paves the way for design of single antagonists directed towards multiple toxins. PMID:27180909

  11. Antimicrobial Resistance of Staphylococcal Strains Isolated from Various Pathological Products

    Directory of Open Access Journals (Sweden)

    Laura-Mihaela SIMON

    2010-12-01

    Full Text Available Background: The optimal choice of antimicrobial therapy is an important problem in hospital environment in which the selection of resistant and virulent strains easy occurs. S. aureus and especially MRSA(methicillin-resistant S. aureus creates difficulties in both treatment and prevention of nosocomial infections. Aim: The purpose of this study is to determine the sensitivity and the resistance to chemotherapy of staphylococci strains isolated from various pathological products. Material and Method: We identified Staphylococccus species after morphological appearance, culture properties, the production of coagulase, hemolisines and the enzyme activity. The susceptibility tests were performed on Mueller-Hinton medium according to CLSI (Clinical and Laboratory Standards Institute. Results: The strains were: MSSA (methicillin-susceptible S. aureus (74%, MRSA (8%, MLS B (macrolides, lincosamides and type B streptogramines resistance (12% and MRSA and MLS B (6%. MRSA strains were more frequently isolated from sputum. MRSA associated with the MLS B strains were more frequently isolated from pus. MLS B strains were more frequently isolated from sputum and throat secretions. All S. aureus strains were susceptible to vancomycin and teicoplanin. Conclusions: All staphylococcal infections require resistance testing before treatment. MLS B shows a high prevalence among strains of S. aureus. The association between MLS B and MRSA remains a major problem in Romania.

  12. Molecular Determinants of Staphylococcal Biofilm Dispersal and Structuring

    Directory of Open Access Journals (Sweden)

    Katherine Y Le

    2014-11-01

    Full Text Available Staphylococci are frequently implicated in human infections, and continue to pose a therapeutic dilemma due to their ability to form deeply seated microbial communities, known as biofilms, on the surfaces of implanted medical devices and host tissues. Biofilm development has been proposed to occur in three stages: 1 attachment, 2 proliferation/structuring, and 3 detachment/dispersal. Although research within the last several decades has implicated multiple molecules in the roles as effectors of staphylococcal biofilm proliferation/structuring and detachment/dispersal, to date, only phenol soluble modulins (PSMs have been consistently demonstrated to serve in this role under both in-vitro and in-vivo settings. PSMs are regulated directly through a density-dependent manner by the accessory gene regulator (Agr system. They disrupt the non-covalent forces holding the biofilm extracellular matrix together, which is necessary for the formation of channels, a process essential for the delivery of nutrients to deeper biofilm layers, and for dispersal/dissemination of clusters of biofilm to distal organs in acute infection. Given their relevance in both acute and chronic biofilm-associated infections, the Agr system and the psm genes hold promise as potential therapeutic targets.

  13. Structural basis of cell wall cleavage by a staphylococcal autolysin.

    Directory of Open Access Journals (Sweden)

    Sebastian Zoll

    2010-03-01

    Full Text Available The major autolysins (Atl of Staphylococcus epidermidis and S. aureus play an important role in cell separation, and their mutants are also attenuated in virulence. Therefore, autolysins represent a promising target for the development of new types of antibiotics. Here, we report the high-resolution structure of the catalytically active amidase domain AmiE (amidase S. epidermidis from the major autolysin of S. epidermidis. This is the first protein structure with an amidase-like fold from a bacterium with a gram-positive cell wall architecture. AmiE adopts a globular fold, with several alpha-helices surrounding a central beta-sheet. Sequence comparison reveals a cluster of conserved amino acids that define a putative binding site with a buried zinc ion. Mutations of key residues in the putative active site result in loss of activity, enabling us to propose a catalytic mechanism. We also identified and synthesized muramyltripeptide, the minimal peptidoglycan fragment that can be used as a substrate by the enzyme. Molecular docking and digestion assays with muramyltripeptide derivatives allow us to identify key determinants of ligand binding. This results in a plausible model of interaction of this ligand not only for AmiE, but also for other PGN-hydrolases that share the same fold. As AmiE active-site mutations also show a severe growth defect, our findings provide an excellent platform for the design of specific inhibitors that target staphylococcal cell separation and can thereby prevent growth of this pathogen.

  14. Pelvic Primary Staphylococcal Infection Presenting as a Thigh Abscess

    Directory of Open Access Journals (Sweden)

    T. O. Abbas

    2013-01-01

    Full Text Available Intra-abdominal disease can present as an extra-abdominal abscess and can follow several routes, including the greater sciatic foramen, obturator foramen, femoral canal, pelvic outlet, and inguinal canal. Nerves and vessels can also serve as a route out of the abdomen. The psoas muscle extends from the twelfth thoracic and fifth lower lumbar vertebrae to the lesser trochanter of the femur, which means that disease in this muscle group can migrate along the muscle, out of the abdomen, and present as a thigh abscess. We present a case of a primary pelvic staphylococcal infection presenting as a thigh abscess. The patient was a 60-year-old man who presented with left posterior thigh pain and fever. Physical examination revealed a diffusely swollen left thigh with overlying erythematous, shiny, and tense skin. X-rays revealed no significant soft tissue lesions, ultrasound was suggestive of an inflammatory process, and MRI showed inflammatory changes along the left hemipelvis and thigh involving the iliacus muscle group, left gluteal region, and obturator internus muscle. The abscess was drained passively via two incisions in the posterior left thigh, releasing large amounts of purulent discharge. Subsequent bacterial culture revealed profuse growth of Staphylococcus aureus. The patient recovered uneventfully except for a moderate fever on the third postoperative day.

  15. Pelvic primary staphylococcal infection presenting as a thigh abscess.

    Science.gov (United States)

    Abbas, T O

    2013-01-01

    Intra-abdominal disease can present as an extra-abdominal abscess and can follow several routes, including the greater sciatic foramen, obturator foramen, femoral canal, pelvic outlet, and inguinal canal. Nerves and vessels can also serve as a route out of the abdomen. The psoas muscle extends from the twelfth thoracic and fifth lower lumbar vertebrae to the lesser trochanter of the femur, which means that disease in this muscle group can migrate along the muscle, out of the abdomen, and present as a thigh abscess. We present a case of a primary pelvic staphylococcal infection presenting as a thigh abscess. The patient was a 60-year-old man who presented with left posterior thigh pain and fever. Physical examination revealed a diffusely swollen left thigh with overlying erythematous, shiny, and tense skin. X-rays revealed no significant soft tissue lesions, ultrasound was suggestive of an inflammatory process, and MRI showed inflammatory changes along the left hemipelvis and thigh involving the iliacus muscle group, left gluteal region, and obturator internus muscle. The abscess was drained passively via two incisions in the posterior left thigh, releasing large amounts of purulent discharge. Subsequent bacterial culture revealed profuse growth of Staphylococcus aureus. The patient recovered uneventfully except for a moderate fever on the third postoperative day.

  16. Skin regeneration in deep second-degree scald injuries either by infusion pumping or topical application of recombinant human erythropoietin gel

    Directory of Open Access Journals (Sweden)

    Giri P

    2015-05-01

    Full Text Available Priya Giri,1 Sabine Ebert,1 Ulf-Dietrich Braumann,2 Mathias Kremer,3 Shibashish Giri,1 Hans-Günther Machens,4 Augustinus Bader1 1Department of Cell Techniques and Applied Stem Cell Biology, Center for Biotechnology and Biomedicine (BBZ, Faculty of Medicine, University of Leipzig, Leipzig, Germany; 2Interdisciplinary Center for Bioinformatics (IZBI, University of Leipzig, Leipzig, Germany; 3Department of Plastic and Hand Surgery, University of Lübeck, Lübeck, Germany; 4Department of Plastic and Hand Surgery, Technical University of Munich, Munich, Germany Abstract: Large doses of recombinant growth factors formulated in solution form directly injected into the body is usual clinical practice in treating second-degree scald injuries, with promising results, but this approach creates side effects; furthermore, it may not allow appropriate levels of the factor to be sensed by the target injured tissue/organ in the specific time frame, owing to complications arising from regeneration. In this research, two delivery methods (infusion pumping and local topical application were applied to deliver recombinant human erythropoietin (rHuEPO for skin regeneration. First, rHuEPO was given in deep second-degree scald injury sites in mice by infusion pump. Vascularization was remarkably higher in the rHuEPO pumping group than in controls. Second, local topical application of rHuEPO gel was given in deep second-degree scald injury sites in rats. Histological analysis showed that epithelialization rate was significantly higher in the rHuEPO gel-treated group than in controls. Immunohistochemical studies showed that the rHuEPO gel-treated group showed remarkably higher expression of skin regeneration makers than the control group. An accurate method for visualization and quantification of blood vessel networks in target areas has still not been developed up to this point, because of technical difficulties in detecting such thin blood vessels. A method which

  17. Application of the antibiotic batumin for accurate and rapid identification of staphylococcal small colony variants

    Directory of Open Access Journals (Sweden)

    Churkina Larisa N

    2012-07-01

    Full Text Available Abstract Background Staphylococcus aureus is a major human pathogen causing significant morbidity and mortality. The S. aureus colonies in osteomyelitis, in patients with cystic fibrosis and patients with endoprosthesis rejection frequently have an atypical morphology, i.e. staphylococcal small-colony variants, which form a naturally occurring subpopulation of clinically important staphylococci. Identification of these small colony variants is difficult, because of the loss of typical phenotypic characteristics of these variants. We wanted to improve and simplify the diagnosis of staphylococcal infection using a diagnostic preparation, consisting of 5 μg batumin paper disks. Batumin possesses a unique selective activity against all studied Staphylococcus spp., whereas all other species tested thus far are batumin resistant. We assessed the efficacy of the batumin diagnostic preparation to identify staphylococcal small colony variants, isolated from osteomyelitis patients. Findings With the batumin diagnostic preparation, all 30 tested staphylococcal small-colony variants had a growth inhibition zone around the disk of minimum 25 mm, accordant with the inhibition zones of the parent strains, isolated from the same patients. Conclusions The batumin diagnostic preparation correctly identified the small-colony variants of S. aureus, S. haemolyticus and S. epidermidis as belonging to the genus Staphylococcus, which differ profoundly from parental strains and are difficult to identify with standard methods. Identification of staphylococcal small-colony variants with the batumin diagnostic preparation is technically simple and can facilitate practical laboratory work.

  18. Monoclonal antibody-based sandwich ELISA for the detection of staphylococcal enterotoxin A.

    Science.gov (United States)

    Kuang, Hua; Wang, Wenbing; Xu, Liguang; Ma, Wei; Liu, Liqiang; Wang, Libing; Xu, Chuanlai

    2013-04-19

    A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated for the detection of staphylococcal enterotoxin A (SEA). After routine fusion and selection, 10 monoclonal antibodies showed high affinity for SEA. An optimal pair for sandwich ELISA was selected by pairwise interaction analysis. After optimization, the limit of detection (LOD) and linear dynamic range of the method were established, and were found to be 0.0282 ng/mL and 0.06-2 ng/mL, respectively. The recovery in pure milk ranged from 82.67% to 111.95% and the intra- and inter-assay coefficients of variation ranged from 3.16% to 6.05% and from 5.16% to 10.79%, respectively. Cross-reactivity with staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), staphylococcal enterotoxin D (SED), and staphylococcal enterotoxin E (SEE) in this method were insignificant. These results indicate that the sandwich ELISA method developed in our study is effective for routine identification of SEA in food samples.

  19. RAGE Deficiency Impairs Bacterial Clearance in Murine Staphylococcal Sepsis, but Has No Significant Impact on Staphylococcal Septic Arthritis.

    Science.gov (United States)

    Mohammad, Majd; Na, Manli; Welin, Amanda; Svensson, Mattias N D; Ali, Abukar; Jin, Tao; Pullerits, Rille

    2016-01-01

    Septic arthritis is a serious joint disease often caused by Staphylococcus aureus (S. aureus). Receptor for Advanced Glycation End products (RAGE) has an important role in several infections. We sought to investigate the role of RAGE in staphylococcal septic arthritis and sepsis in mice. Wild-type (WT) and RAGE deficient (RAGE-/-) mice were intra-articularly or intravenously inoculated with an arthritic or septic dose of S. aureus LS-1 strain. Clinical arthritis, weight development and mortality were monitored for 14 days. Serum levels of cytokines, kidney bacterial loads as well as micro-CT and histopathology of the joints were assessed. RAGE-/- mice with septic arthritis had significantly lower IL-17A and higher bone mineral density (BMD) compared to the control group. However, no significant differences between the groups were observed regarding the weight loss, the severity and frequency of arthritis, and bacterial loads in the kidneys. In mice with sepsis, the overall mortality rate was similar in RAGE-/- (39%) and in WT mice (45%). However, RAGE-/- mice with sepsis had significantly higher bacterial load in their kidneys compared to the WT controls. In line with data from hematogenous S. aureus arthritis, RAGE deficiency had no impact on arthritis severity in local joint infection. Our results indicate that lack of RAGE has no significant impact on septic arthritis. However, RAGE-/- mice had significantly higher BMD compared to WT mice, which coincided with lower IL-17A in RAGE-/- mice. In sepsis, RAGE deficiency impairs bacterial kidney clearance.

  20. Cross-Talk between Staphylococcus aureus and Other Staphylococcal Species via the agr Quorum Sensing System

    DEFF Research Database (Denmark)

    Canovas de la Nuez, Jaime; Baldry, Mara; Bojer, Martin S;

    2016-01-01

    between staphylococci and S. aureus, and show that this interaction may eventually lead to the identification of new anti-virulence candidates to target S. aureus infections. Here we show that culture supernatants of 37 out of 52 staphylococcal isolates representing 17 different species inhibit S. aureus......-inducing peptides (AIPs) sensed by AgrC, a two component histidine kinase. agr loci are found also in other staphylococcal species and for Staphylococcus epidermidis, the encoded AIP represses expression of agr regulated virulence genes in S. aureus. In this study we aimed to better understand the interaction....... To assess impact on S. aureus virulence, we co-inoculated S. aureus and S. schleiferi in vivo in the Galleria mellonella wax moth larva, and found that expression of key S. aureus virulence factors was abrogated. Our data show that the S. aureus agr locus is highly responsive to other staphylococcal species...

  1. Chromatofocusing: a new method for purification of staphylococcal enterotoxins B and C1.

    Science.gov (United States)

    Ende, I A; Terplan, G; Kickhöfen, B; Hammer, D K

    1983-12-01

    A new chromatographic procedure was developed which obtained highly purified preparations of staphylococcal enterotoxins B and C1 in yields of 60% from cultures of Staphylococcus aureus and which is faster than any of the separation methods used previously. The procedure involves chromatography on carboxymethylcellulose, removal of alpha-toxin by adsorption to rabbit erythrocyte membranes, and finally, chromatofocusing as the fundamental new step. Enterotoxins were obtained in highly purified form and behaved in a homogeneous manner as determined by ultracentrifugation and electrophoresis on polyacrylamide gel in the presence of sodium dodecyl sulfate, with molecular weights of 34,000 for staphylococcal enterotoxin B and 30,000 for staphylococcal enterotoxin C1. Using chromatofocusing as the final purification step, we isolated three B and six C1 distinct but immunologically identical enterotoxin fractions, which were found to be devoid of any impurities and to possess a marked degree of toxicity in monkeys.

  2. Chromatofocusing: a new method for purification of staphylococcal enterotoxins B and C1.

    Science.gov (United States)

    Ende, I A; Terplan, G; Kickhöfen, B; Hammer, D K

    1983-01-01

    A new chromatographic procedure was developed which obtained highly purified preparations of staphylococcal enterotoxins B and C1 in yields of 60% from cultures of Staphylococcus aureus and which is faster than any of the separation methods used previously. The procedure involves chromatography on carboxymethylcellulose, removal of alpha-toxin by adsorption to rabbit erythrocyte membranes, and finally, chromatofocusing as the fundamental new step. Enterotoxins were obtained in highly purified form and behaved in a homogeneous manner as determined by ultracentrifugation and electrophoresis on polyacrylamide gel in the presence of sodium dodecyl sulfate, with molecular weights of 34,000 for staphylococcal enterotoxin B and 30,000 for staphylococcal enterotoxin C1. Using chromatofocusing as the final purification step, we isolated three B and six C1 distinct but immunologically identical enterotoxin fractions, which were found to be devoid of any impurities and to possess a marked degree of toxicity in monkeys. Images PMID:6660872

  3. LYSIS OF BACTERIAL PROTOPLASTS AND SPHEROPLASTS BY STAPHYLOCOCCAL ALPHA-TOXIN AND STREPTOLYSIN S.

    Science.gov (United States)

    BERNHEIMER, A W; SCHWARTZ, L L

    1965-05-01

    Bernheimer, Alan W. (New York University School of Medicine, New York, N.Y.), and Lois L. Schwartz. Lysis of bacterial protoplasts and spheroplasts by staphylococcal alpha-toxin and streptolysin S. J. Bacteriol. 89:1387-1392. 1965.-Protoplasts of Bacillus megaterium, Sarcina lutea, and Streptococcus pyogenes, and spheroplasts of Escherichia coli were lysed by staphylococcal alpha-toxin, whereas spheroplasts of Vibrio metschnikovii and V. comma were not. In the spectrum of its lytic action, streptolysin S qualitatively resembled staphylococcal alpha-toxin except for failure to lyse S. pyogenes protoplasts. In contrast to the two foregoing agents, streptolysin O did not lyse protoplasts and spheroplasts. The observations are interpreted in relation to similarities and differences in lipid composition of bacterial and mammalian cell membranes.

  4. Staphylococcal Superantigens Spark Host-Mediated Danger Signals

    Directory of Open Access Journals (Sweden)

    Terry eKrakauer

    2016-02-01

    Full Text Available Staphylococcal enterotoxin B (SEB of Staphylococcus aureus, and related superantigenic toxins produced by myriad microbes, are potent stimulators of the immune system causing a variety of human diseases from transient food poisoning to lethal toxic shock. These protein toxins bind directly to specific V regions of T-cell receptors (TCR and major histocompatibility complex (MHC class II on antigen-presenting cells, resulting in hyperactivation of T lymphocytes and monocytes / macrophages. Activated host cells produce excessive amounts of proinflammatory cytokines and chemokines, especially tumor necrosis factor α, interleukin 1 (IL-1, IL-2, interferon γ (IFNγ, and macrophage chemoattractant protein 1 causing clinical symptoms of fever, hypotension, and shock. Because of superantigen-induced T cells skewed towards TH1 helper cells, and the induction of proinflammatory cytokines, superantigens can exacerbate autoimmune diseases. Upon TCR / MHC ligation, pathways induced by superantigens include the mitogen-activated protein kinase cascades and cytokine receptor signaling, resulting in activation of NFκB and the phosphoinositide 3-kinase / mammalian target of rapamycin pathways. Various mouse models exist to study SEB-induced shock including those with potentiating agents, transgenic mice and an SEB-only model. However, therapeutics to treat toxic shock remain elusive as host response genes central to pathogenesis of superantigens have only been identified recently. Gene profiling of a murine model for SEB-induced shock reveals novel molecules upregulated in multiple organs not previously associated with SEB-induced responses. The pivotal genes include intracellular DNA / RNA sensors, apoptosis / DNA damage-related molecules, immunoproteasome components, as well as anti-viral and IFN-stimulated genes. The host-wide induction of these, and other, anti-microbial defense genes provide evidence that SEB elicits danger signals resulting in multi

  5. Staphylococcal biofilm exopolysaccharide protects against Caenorhabditis elegans immune defenses.

    Directory of Open Access Journals (Sweden)

    Jakob Begun

    2007-04-01

    Full Text Available Staphylococcus epidermidis and Staphylococcus aureus are leading causes of hospital-acquired infections that have become increasingly difficult to treat due to the prevalence of antibiotic resistance in these organisms. The ability of staphylococci to produce biofilm is an important virulence mechanism that allows bacteria both to adhere to living and artificial surfaces and to resist host immune factors and antibiotics. Here, we show that the icaADBC locus, which synthesizes the biofilm-associated polysaccharide intercellular adhesin (PIA in staphylococci, is required for the formation of a lethal S. epidermidis infection in the intestine of the model nematode Caenorhabditis elegans. Susceptibility to S. epidermidis infection is influenced by mutation of the C. elegans PMK-1 p38 mitogen-activated protein (MAP kinase or DAF-2 insulin-signaling pathways. Loss of PIA production abrogates nematocidal activity and leads to reduced bacterial accumulation in the C. elegans intestine, while overexpression of the icaADBC locus in S. aureus augments virulence towards nematodes. PIA-producing S. epidermidis has a significant survival advantage over ica-deficient S. epidermidis within the intestinal tract of wild-type C. elegans, but not in immunocompromised nematodes harboring a loss-of-function mutation in the p38 MAP kinase pathway gene sek-1. Moreover, sek-1 and pmk-1 mutants are equally sensitive to wild-type and icaADBC-deficient S. epidermidis. These results suggest that biofilm exopolysaccharide enhances virulence by playing an immunoprotective role during colonization of the C. elegans intestine. These studies demonstrate that C. elegans can serve as a simple animal model for studying host-pathogen interactions involving staphylococcal biofilm exopolysaccharide and suggest that the protective activity of biofilm matrix represents an ancient conserved function for resisting predation.

  6. Late multiple organ surge in interferon-regulated target genes characterizes staphylococcal enterotoxin B lethality.

    Directory of Open Access Journals (Sweden)

    Gabriela A Ferreyra

    Full Text Available BACKGROUND: Bacterial superantigens are virulence factors that cause toxic shock syndrome. Here, the genome-wide, temporal response of mice to lethal intranasal staphylococcal enterotoxin B (SEB challenge was investigated in six tissues. RESULTS: The earliest responses and largest number of affected genes occurred in peripheral blood mononuclear cells (PBMC, spleen, and lung tissues with the highest content of both T-cells and monocyte/macrophages, the direct cellular targets of SEB. In contrast, the response of liver, kidney, and heart was delayed and involved fewer genes, but revealed a dominant genetic program that was seen in all 6 tissues. Many of the 85 uniquely annotated transcripts participating in this shared genomic response have not been previously linked to SEB. Nine of the 85 genes were subsequently confirmed by RT-PCR in every tissue/organ at 24 h. These 85 transcripts, up-regulated in all tissues, annotated to the interferon (IFN/antiviral-response and included genes belonging to the DNA/RNA sensing system, DNA damage repair, the immunoproteasome, and the ER/metabolic stress-response and apoptosis pathways. Overall, this shared program was identified as a type I and II interferon (IFN-response and the promoters of these genes were highly enriched for IFN regulatory matrices. Several genes whose secreted products induce the IFN pathway were up-regulated at early time points in PBMCs, spleen, and/or lung. Furthermore, IFN regulatory factors including Irf1, Irf7 and Irf8, and Zbp1, a DNA sensor/transcription factor that can directly elicit an IFN innate immune response, participated in this host-wide SEB signature. CONCLUSION: Global gene-expression changes across multiple organs implicated a host-wide IFN-response in SEB-induced death. Therapies aimed at IFN-associated innate immunity may improve outcome in toxic shock syndromes.

  7. Panton-Valentine Leukocidin-Producing Staphylococcal aureus: Report of Four Siblings.

    LENUS (Irish Health Repository)

    2012-01-31

    Panton-Valentine leukocidin (PVL)-producing Staphylococcus aureus results in leukocyte destruction and tissue necrosis (Pediatric Dermatology 2007;24:401). It can be associated with a spectrum of clinical manifestations that range from localized staphylococcal skin infections to sometimes severe necrotizing pneumonia (Clin Infect Dis 1999;29:1128). We report a case of four siblings, three brothers whose atopic dermatitis was complicated by cutaneous lesions and furunculosis, while their 21-month-old sister had a fatal PVL positive staphylococcal pneumonia.

  8. Protective effect of z-VAD-fmk on kidney in rats after-scald%广谱caspase抑制剂对大鼠烫伤后肾脏的保护作用

    Institute of Scientific and Technical Information of China (English)

    朱宝昌; 盛小辉; 刘群; 李红卫

    2014-01-01

    point mentioned above. Renal morphology dyed with HE was observed with microscope. Detection of apoptotic renal cells was carried out using TUNEL and activity of caspase-3 was measured with colorimetric chro-matometry. Results Compared with the injury group,the damage of rat renal cells were lessened in treatment group,the apop-tosis index (AI) of renal cells significantly decreased at 8h,24h and 48h (P<0.01) the change of caspase-3 activity at 2h, 8h, 24h and 48h significantly decreased after scald (P<0.01). Conclusion The renal cells apoptosis participated in renal injury after-scald. The early intervention of z-VAD-fmk could effectively reduce renal cells apoptosis and relieve kidney damage.

  9. Anti-staphylococcal effectiveness of nisaplin in refrigerated pizza doughs

    Directory of Open Access Journals (Sweden)

    Wilma Carla de Freitas

    2008-06-01

    Full Text Available This study evaluated the effectiveness of nisaplin, commercial product having nisin as active component, in decreasing the staphylococcal population in refrigerated pizza doughs. The refrigerated pizza dough pieces randomly chosen were dipped in the solutions with nisaplin concentrations of 1.0 x 10-3 g and 1.0 x 10-2 g nisaplin/mL named for the treatment A and B and kept under refrigeration (7 °C ± 1 °C. On times 0, 15 and 30 days post treatment the Staphylococcus spp. count was carried out. The results showed that both nisaplin treatments were able to reduce the Staphylococcus spp. count (CFU/g in the refrigerated pizza doughs. However, only treatment B showed statistically significant reducer effect (p Este estudo avaliou a efetividade de nisaplin, produto comercial tendo nisina como componente ativo, em diminuir a população estafilocócica em massas de pizza refrigeradas. Pedaços de massa de pizza refrigerada foram randomicamente escolhidos e mergulhados em soluções de nisaplin com concentração de 1.0 10-3 g e 1.0 10-2 g nisaplin/mL, nomeados, respectivamente, tratamento A e B, e mantidos sob refrigeração (7 °C ± 1°C. Nos tempos 0, 15 e 30 dias pós-tratamento foram feitas as contagens de Staphylococcus spp. Os resultados mostraram que ambos os tratamentos diminuíram a contagem (UFC/g de Staphylococcus spp. em massas de pizza refrigeradas. Entretanto, somente o tratamento B mostrou um efeito redutor estatisticamente significante (p < 0.05 sobre a contagem de Staphylococcus spp. causando uma diminuição de 1.0 e 0.98 ciclos logarítmicos, respectivamente, nos tempos 15 e 30 dias pós tratamento. Estes dados sugerem que nisina poderia apresentar-se como uma promissora alternativa para controlar o a sobrevivência de microrganismos patógenos em alimentos, particularmente, espécies de Staphylococcus em massas de pizza refrigerada.

  10. 双黄烧伤膏对大鼠烧伤、烫伤的治疗作用研究%Experimental studies of shuanghuang burn ointment treatment for scald and burn wound of rat model

    Institute of Scientific and Technical Information of China (English)

    朱晓红; 凌磊; 王杨; 王强

    2012-01-01

    目的 研究双黄烧伤膏对大鼠实验性烫伤、烧伤感染的治疗作用.方法 将大鼠随机分为实验组(双黄烧伤膏组)、阳性对照组(京万红软膏组)、空白对照组.建立大鼠Ⅱ度烫伤模型、Ⅲ度烧伤感染模型,分别涂布给药,单笼饲养,观察并记录创面变化.结果 在大鼠烫伤实验中,给药15 d后,各组创面结痂面积明显变小,实验组结痂愈合时间短,与阳性对照组、空白对照组比较,差异有统计学意义(P0.05),与空白对照组相比,差异有统计学意义(P0.05),与阴性对照组相比,差异有统计学意义(P<0.01).结论 双黄烧伤膏对大鼠实验性烫伤、烧伤感染具有较好的治疗作用.%Objective To study the effects of shuanghuang burn ointment on scald and burned infectious rats. Methods Rats were randomly divided into test group( shuanghuang burn ointment ), positive control group( jingwan-hong cream )and no treatment group respectively. The drugs were administered topically on the surface of scalded and burned infection places respectively after partical-thickness scald and burn in rats. The change of the scalded and burned areas and the decrustation time were recorded. Results In the partical-thickness scalding test,the scalded areas of 15 d were smaller, there was significant difference between test group and positive control group( P < 0. 05 ), there was significant difference between test group and no treatment group( P < 0. 01 ); the scalded areas of 20 d: there was significant difference between test group and no treatment group( P <0. 01 ),but there was no significant difference compared with the positive control group. In the partical-thickness burnning infection test,the scalded areas of 9 d,there was significant difference between the test group and the no treatment group( P <0. 01 ) ,but there was no significant difference compared with the positive control group. Conclusion Burn ointment has a good anti-inflammatory, antibacterial and

  11. Suppressive therapy using azithromycin in 2 rare cases of recurrent staphylococcal infections.

    Science.gov (United States)

    Grobost, Vincent; Rigal, Emilie; Pavier, Yoann; Vidal, Magali; Mrozek, Natacha; Beytout, Jean; Laurichesse, Henri; Lesens, Olivier

    2014-05-01

    Recurrent staphylococcal skin and soft tissue infections may recur despite decontamination and multiple courses of antibiotic therapy and may dramatically impair the patient's quality of life. We report successful use of long-term azithromycin prophylaxis in a recurrent laryngitis and a scalp folliculitis due to methicillin-susceptible Staphylococcus aureus.

  12. Occurrence of Staphylococcal Ocular Infections of Food Producing Animals in Nsukka Southeast, Nigeria

    Directory of Open Access Journals (Sweden)

    Sunday Ositadinma Udegbunam

    2014-01-01

    Full Text Available Staphylococcal ocular infections of food animals have been somewhat under diagnosed probably due to the ubiquitous nature of staphylococcal organisms. This study was undertaken to determine the occurrence of staphylococcal ocular infections of food producing animals in Nsukka Southeast, Nigeria, and to determine the antibiogram of the isolated staphylococci. A total of 5,635 food producing animals were externally examined for signs of clinical ocular conditions. Animals that showed clinical eye lesions were further examined using pen light to assess the entire globe and the pupillary reflex. Blindness was assessed using menace blink reflex, palpebral reflex and obstacle methods. Isolation and identification of staphylococcal isolates from ocular swabs were done by standard methods. Antibiogram of the isolates was determined by disc diffusion method. Sixty-three (1.1% of the examined animals showed signs of ocular condition. Thirty-one (49.2% of the cultured swabs yielded Staphylococcus aureus (S. aureus. Isolation rates from different animal species were caprine (60%, ovine (33.3%, bovine (12.5%, and porcine (0%. Resistance of the isolates was 100% to ampicillin/cloxacillin, 90% to tetracycline, 80% to streptomycin, 71% to chloramphenicol, 20% to erythromycin, 16% to gentamicin, and 0% to ciprofloxacin and norfloxacin. Twenty-five (81% of the isolates were multi-drug resistant. This study has shown that antibiotic-resistant staphylococci are associated with a sizeable percentage of ocular infections of food producing animals and should be considered during diagnosis and treatment.

  13. In vitro cell based assay for activity analysis of staphylococcal enterotoxin A in food

    Science.gov (United States)

    Staphylococcal enterotoxins (SEs) are a leading cause of food poisoning. They function both as toxins that cause gastroenteritis after ingestion and as superantigens that non-specifically activate large numbers of T cells. Monkey or kitten bioassays were historically developed for analysis of SE act...

  14. The olive compound 4-hydroxytyrosol inactivates Staphyloccoccus aureus bacteria and Staphylococcal enterotoxin A (SEA)

    Science.gov (United States)

    The foodborne pathogen Staphylococcus aureus produces the virulent staphylococcal enterotoxin A (SEA), a single chain protein which consists of 233 amino acid residues with a molecular weight of 27,078 Da. SEA is a superantigen that is reported to contribute to animal (mastitis) and human (emesis, ...

  15. Inhibition of Biological Activity of Staphylococcal Enterotoxin A (SEA) by Apple Juice and Apple Polyphenols

    Science.gov (United States)

    The foodborne pathogen Staphylococcus aureus produces the virulent staphylococcal entertoxin A (SEA), a single-chain protein that consists of 233 amino acid residues with a molecular weight of 27 078 Da. SEA is a superantigen that is reported to contribute to animal (mastitis) and human (emesis, dia...

  16. Anti-staphylococcal humoral immune response in persistent nasal carriers and noncarriers of Staphylococcus aureus

    NARCIS (Netherlands)

    N.J. Verkaik (Nelianne); C.P. de Vogel (Corné); H.A.M. Boelens (Hélène); D. Grumann (Dorothee); T. Hoogenboezem (Theo); C. Vink (Cornelis); H. Hooijkaas (Herbert); T.J. Foster (Timothy); H.A. Verbrugh (Henri); A.F. van Belkum (Alex); W.J.B. van Wamel (Willem)

    2009-01-01

    textabstractBACKGROUND. Persistent carriers have a higher risk of Staphylococcus aureus infections than noncarriers but a lower risk of bacteremia-related death. Here, the role played by anti-staphylococcal antibodies was studied. METHODS. Serum samples from 15 persistent carriers and 19 noncarriers

  17. Assessment of a microplate method for detection of staphylococcal secretory inhibitor of platelet microbicidal protein.

    Science.gov (United States)

    Ivanov, Iuri B; Gritsenko, Viktor A

    2009-01-01

    We developed a novel microplate spectrophotometric assay for the detection of staphylococcal secretory inhibitor of platelet microbicidal protein (SIPMP) in

  18. Staphylococcal secretory inhibitor of platelet microbicidal protein is associated with prostatitis source.

    Science.gov (United States)

    Ivanov, Iuri B; Gritsenko, Viktor A; Kuzmin, Michael D

    2006-12-01

    This study reports the detection of an extracellular staphylococcal product, designated secretory inhibitor of platelet microbicidal protein (SIPMP), that causes local inhibition of the bactericidal action of platelet microbicidal protein (PMP) in the fluid phase. Urethral isolates of Staphylococcus aureus (n=24) and coagulase-negative staphylococci (CNS) (n=47) from patients with or without chronic bacterial prostatitis (CBP) were tested. SIPMP production was tested by inhibition of PMP bioactivity against Bacillus subtilis and was expressed as percentage inhibition of PMP bactericidal activity. The PMP susceptibility of staphylococcal strains was determined by exposing bacterial cells to serial dilutions of PMP. Staphylococci from patients without CBP produced SIPMP at levels of 10.3+/-1.2 and 13.25+/-1.72 % for S. aureus and CNS, respectively. Strains isolated from men with CBP inhibited PMP-induced killing of B. subtilis by 23.38+/-4.2 % (P<0.05) and 23.69+/-1.87 % (P<0.01) for S. aureus and CNS, respectively. SIPMP production correlated with staphylococcal resistance to PMP (r2=0.6082 and 0.7264 for S. aureus and CNS, respectively). SIPMP represents a hitherto unrecognized determinant of staphylococcal pathogenicity. These results suggest that SIPMP production is associated with the CBP source. Data from this study may have significant implications for the understanding of the pathogenesis of CBP.

  19. Rhodomyrtus tomentosa (Aiton) Hassk. leaf extract: An alternative approach for the treatment of staphylococcal bovine mastitis.

    Science.gov (United States)

    Mordmuang, Auemphon; Voravuthikunchai, Supayang Piyawan

    2015-10-01

    Antibiotic residues in dairy products as well as emergence of antimicrobial resistance in foodborne pathogens have been recognized as global public health concerns. The present work was aimed to study a potent antibacterial extract from natural product as an alternative treatment for staphylococcal bovine mastitis. Staphylococcal isolates (n=44) were isolated from milk samples freshly squeezed from individual cows. All staphylococcal isolates were resistant to ampicillin, ciprofloxacin, erythromycin, gentamicin, penicillin, except vancomycin. Rhodomyrtus tomentosa leaf ethanolic extract was accessed for its antibacterial activity and anti-inflammatory potential. The extract exhibited profound antibacterial activity against all of staphylococcal isolates with MIC and MBC values ranged from 16-64 μg/ml and 64->128 μg/ml, respectively. Moreover, the extract also exerted anti-protein denaturation and human red blood cell membrane stabilizing activity. The results support the use of R. tomentosa extract that could be applied to cure bovine mastitis and to reduce inflammatory injury caused by the bacterial infections. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Staphylococcal SSL5 binding to human leukemia cells inhibits cell adhesion to endothelial cells and platelets

    NARCIS (Netherlands)

    Walenkamp, Annemiek M. E.; Bestebroer, Jovanka; Boer, Ingrid G. J.; Kruizinga, Roeline; Verheul, Henk M.; van Strijp, Jos A. G.; de Haas, Carla J. C.

    2010-01-01

    Bacterial proteins provide promising tools for novel anticancer therapies. Staphylococcal superantigen-like 5 (SSL5) was recently described to bind P-selectin glycoprotein ligand-1 (PSGL-1) on leukocytes and to inhibit neutrophil rolling on a P-selectin surface. As leukocytes and tumor cells share m

  1. Lab-on-a-chip for label free biological semiconductor analysis of Staphylococcal Enterotoxin B

    NARCIS (Netherlands)

    Yang, Minghui; Sun, Steven; Bruck, Hugh Alan; Kostov, Yordan; Rasooly, Avraham

    2010-01-01

    We describe a new lab-on-a-chip (LOC) which utilizes a biological semiconductor (BSC) transducer for label free analysis of Staphylococcal Enterotoxin B (SEB) (or other biological interactions) directly and electronically. BSCs are new transducers based on electrical percolation through a multi-laye

  2. Mobile and immobile adhesion of staphylococcal strains to hydrophilic and hydrophobic surfaces

    NARCIS (Netherlands)

    Boks, Niels P.; Kaper, Hans J.; Norde, Willem; van der Mei, Henny C.; Busscher, Henk J.

    2009-01-01

    Staphylococcus epidermidis adheres to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in similar numbers, but in different modes. Real-time observation of staphylococcal adhesion under a shear rate of 15 s(-1) revealed different adhesion dynamics on both substrata. The nu

  3. Temporal expression of staphylococcal enterotoxin h in comparison with accessory gene regulator-dependent and -independent enterotoxins.

    Science.gov (United States)

    Lis, Elżbieta; Podkowik, Magdalena; Bystroń, Jarosław; Stefaniak, Tadeusz; Bania, Jacek

    2012-02-01

    Using sandwich enzyme-linked immunosorbent assay (ELISA), the production of staphylococcal enterotoxin (SE) H was determined in 22 Staphylococcus aureus isolates bearing the seh gene. Samples of supernatants were taken at four time points corresponding to exponential phase (optical density at 600 nm [OD(600)] 0.3 to 0.6), late exponential phase (OD(600) 2 to 4), early stationary phase (OD(600) 4 to 6), and late stationary phase (OD(600) 7 to 12). In four isolates, SEH was detectable at a very low level at the first time point. In 18 isolates, the earliest SEH production was detected in the late exponential phase. For all isolates, there was an increase of SEH concentration with time. Western blot analysis revealed that SEH production, similar to SEA, started in the early exponential phase (OD(600) ∼ 0.5). Isolates with high SEH productivity, as measured by ELISA, demonstrated a higher seh transcription as well. sec transcription was induced in the stationary phase. An induction in the sea transcript was observed during mid- to late exponential phase. Expression profile of seh was similar to that of sea. We showed that the seh expression profile is similar to that of Agr-independent sea and not to that of Agr-dependent sec genes. SEH can be effectively expressed at low bacterial counts, meaning that even in an environment not favorable for S. aureus growth, seh-bearing strains can pose a risk for food safety.

  4. A novel electrochemical immunosensor based on magnetosomes for detection of staphylococcal enterotoxin B in milk.

    Science.gov (United States)

    Wu, Longyun; Gao, Bo; Zhang, Fang; Sun, Xiulan; Zhang, Yinzhi; Li, Zaijun

    2013-03-15

    In this paper, a novel electrochemical immunosensor to detect staphylococcal enterotoxin B based on bio-magnetosomes, polyaniline nano-gold composite and 1,2-dimethyl-3-butylimidazolium hexafluorophosphate ionic liquid, was developed, and found to exhibit high sensitivity and stability. The specific antibody to staphylococcal enterotoxin B conjugated with the magnetosomes showed rapid immunoreactions and good dispersion, which contributed to the formation of a nanostructurally smooth and dense film on the surface of a gold electrode. Polyaniline nano-gold composite and 1,2-dimethyl-3-butylimidazolium hexafluorophosphate ionic liquid were used to modify the electrode as mediators to improve the electron transfer and offer an excellent biocompatible microenvironment for the antibody to retain its activity to enhance the response of the electrochemical sensor. Under optimal conditions, the developed immunosensor showed a good linear response in the range from 0.05 to 5 ng/mL (R(2)=0.9957) with a detection limit as low as 0.017 ng/mL, compared with the one without magnetosomes (0.05-5 ng/mL, 0.033 ng/mL), this developed immunosensor showed a wider response range and a reduced detection limit. And a good specificity with little adsorption to staphylococcal enterotoxin A, C and Na(+), K(+), Ca(2+) was obtained. Moreover, the immunosensor exhibited a good long-time stability at 4 °C reaching up to 60 days, which showed a relatively long working life. Meanwhile the immunosensor could be regenerated four times using NaOH elution. The sensor also displayed a good repeatability with a relative standard deviation of 5.02% for staphylococcal enterotoxin B detection (1 ng/mL, n=9). Furthermore, high recoveries in milk samples from 81% to 118% were achieved and successfully applied to milk sample detection. The obtained results demonstrate that the developed electrochemical immunosensor is a promising tool for the detection of staphylococcal enterotoxin B in food.

  5. Inhibition of Exotoxin Production by Mobile Genetic Element SCCmec-Encoded psm-mec RNA Is Conserved in Staphylococcal Species

    OpenAIRE

    Mariko Ikuo; Gentaro Nagano; Yuki Saito; Han Mao; Kazuhisa Sekimizu; Chikara Kaito

    2014-01-01

    Staphylococcal species acquire antibiotic resistance by incorporating the mobile-genetic element SCCmec. We previously found that SCCmec-encoded psm-mec RNA suppresses exotoxin production as a regulatory RNA, and the psm-mec translation product increases biofilm formation in Staphylococcus aureus. Here, we examined whether the regulatory role of psm-mec on host bacterial virulence properties is conserved among other staphylococcal species, S. epidermidis and S. haemolyticus, both of which are...

  6. Crystal Structure of Staphylococcal Enterotoxin G (SEG) in Complex with a Mouse T-cell Receptor Beta Chain

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez, M.M.; Robinson, H.; Cho, S.; De Marzi, M. C.; Kerzic, M. C.; Mariuzza, R. A.; Malchiodi, E. L.

    2011-01-14

    Superantigens (SAgs) are bacterial or viral toxins that bind MHC class II (MHC-II) molecules and T-cell receptor (TCR) in a nonconventional manner, inducing T-cell activation that leads to inflammatory cytokine production, which may result in acute toxic shock. In addition, the emerging threat of purpura fulminans and community-associated meticillin-resistant Staphylococcus aureus emphasizes the importance of a better characterization of SAg binding to their natural ligands that may allow the development of reagents to neutralize their action. The three-dimensional structure of the complex between a mouse TCR {beta} chain (mV{beta}8.2) and staphylococcal enterotoxin G (SEG) at 2.0 {angstrom} resolution revealed a binding site that does not conserve the 'hot spots' present in mV{beta}8.2-SEC2, mV{beta}8.2-SEC3, mV{beta}8.2-SEB, and mV{beta}8.2-SPEA complexes. Analysis of the mV{beta}8.2-SEG interface allowed us to explain the higher affinity of this complex compared with the others, which may account for the early activation of T-cells bearing mV{beta}8.2 by SEG. This mode of interaction between SEG and mV{beta}8.2 could be an adaptive advantage to bestow on the pathogen a faster rate of colonization of the host.

  7. Plaque identification of strand-forming canine distemper virus by staphylococcal protein A-mediated reverse passive haemadsorption.

    Science.gov (United States)

    Johnson, G C; Fulks, K; Krakowka, S

    1985-02-01

    The R252 neurotropic isolate of canine distemper virus (CDV) produces cytopathic effects (CPE) dominated by strand formation rather than by the formation of multinucleate giant cells. The lack of well-defined CPE and consequent rapid spread of infection throughout the cell monolayer has hindered plaque purification of this virus by conventional methods. However, the use of an immunological detection system which utilizes binding of hyperimmune dog serum to virus-infected cells, followed by the identification of those sites by staphylococcal Protein A-coupled sheep red blood cells (reverse passive haemadsorption) allowed infected foci in cell monolayers to be detected as early as 4 days after infection, coincident with the appearance of the first immunofluorescently identified viral foci. Foci of haemadsorption were specific to sites of CDV infection as demonstrated by blocking experiments. Material recovered from the plaques was successful in infecting Vero cells. Thus, immunologically mediated adsorption of Protein A coupled red blood cells can be used to identify and isolate foci of viral infection which exhibit minimal or no viral CPE without destroying viral replicative ability.

  8. Preservation Agent and Inhibitory Effects of Resveratrol on Superficial Scald of Dangshan Pear%白藜芦醇对砀山酥梨的保鲜效应

    Institute of Scientific and Technical Information of China (English)

    金宏; 惠伟; 牛瑞雪

    2014-01-01

    文章以砀山酥梨果实为试材,在果实冷藏(2士0.5℃)前,采用0.5μL·L-1白藜芦醇溶液浸泡2 min,定期对果实保鲜过程中相关的生理指标进行测定,在贮藏后期对梨的黑皮病发病情况做统计,研究白藜芦醇对砀山酥梨果实采后保鲜效应和黑皮病的抑制及可能发病机理.结果表明:白藜芦醇处理对砀山酥梨有较好的保鲜效果,可抑制共轭三烯的含量、增加细胞的膜透性、降低α-法尼烯的峰值,显著抑制和延缓梨果实在贮藏期和货架期黑皮病的发生.%Pear fruits( Dangshan pear)were treated with resveratrol at 0.5 μL·L-1 for 2 min before cold stor-age(2 ± 0.5)℃. Some indexes of post-harvest physiology were researched and the complexion of superficial scald were observed.The inhibitory effects and possible mechanisms of resveratrol on superficial scald of Dangshan pear were studied.The experimental results show that resveratrol at 0.5 μL · L-1 could inhibited the content of α-farnesene,conjugated trienes,decreased the relative membrane permeability and kept the Dangshan pears relatively fresh during cold storage.The appropriate concentrations of resveratrol could re-strain and defer the occurrence of superficial scald of Dangshan pear remarkably in cold storage period and shelf life respectively.

  9. Therapeutic Inhibition of Pro-Inflammatory Signaling and Toxicity to Staphylococcal Enterotoxin B by a Synthetic Dimeric BB-Loop Mimetic of MyD88

    Science.gov (United States)

    2012-07-27

    Therapeutic Inhibition of Pro-Inflammatory Signaling and Toxicity to Staphylococcal Enterotoxin B by a Synthetic Dimeric BB-Loop Mimetic of MyD88...Maryland, United States of America Abstract Staphylococcal enterotoxin B (SEB) exposure triggers an exaggerated pro-inflammatory cytokine response...Therapeutic Inhibition of Pro-Inflammatory Signaling and Toxicity to Staphylococcal Enterotoxin B by a Synthetic Dimeric BB-Loop Mimetic of MyD88

  10. Therapeutic Down-Modulators of Staphylococcal Superantigen-Induced Inflammation and Toxic Shock

    Directory of Open Access Journals (Sweden)

    Teresa Krakauer

    2010-07-01

    Full Text Available Staphylococcal enterotoxin B (SEB and related superantigenic toxins are potent stimulators of the immune system and cause a variety of diseases in humans, ranging from food poisoning to toxic shock. These toxins bind directly to major histocompatibility complex (MHC class II molecules on antigen-presenting cells and specific Vb regions of T-cell receptors (TCR, resulting in hyperactivation of both monocytes/macrophages and T lymphocytes. Activated host cells produce massive amounts of proinflammatory cytokines and chemokines, activating inflammation and coagulation, causing clinical symptoms that include fever, hypotension, and shock. This review summarizes the in vitro and in vivo effects of staphylococcal superantigens, the role of pivotal mediators induced by these toxins in the pathogenic mechanisms of tissue injury, and the therapeutic agents to mitigate the toxic effects of superantigens.

  11. In vitro construction of tissue engineered skin for wound repair after escharectomy of third degree scald: An experimental study

    Directory of Open Access Journals (Sweden)

    Zhong-feng MA

    2016-01-01

    Full Text Available Objective  To observe the practicability and effect of tissue engineered skin for repairing the wound after escharectomy of third degree scald (TDSE in rat model. Methods  Epithelial cells and fibroblasts from newborn SD rats were isolated by enzyme digestion method and cultured in vitro, and porcine acellular dermal matrix (PADM without cytotoxicity was prepared by hyperosmotic saline/sodium hydroxide method. The fibroblasts were mixed with bovine type Ⅰ collagen and inoculated on the surface of PADM. Third passage of cultured epidermal cells from newborn SD rats were inoculated on the collagen surface of the dermal matrix to obtain tissue engineered skin, and it was used to prepare epidermal cell sheet. Forty-eight SD rats with TDSE wound were randomly divided into two groups, then tissue engineered skin (experiment group, and epidermal cell sheet (control group graftings were performed to cover the wounds respectively. Finally, gross observation and histological changes were observed in grafted area. The wound healing rate and wound contraction rate were compared between the two groups. Microvessel count (MVC was performed with antiCD34 monoclonal antibody immunohistochemical staining technique, and vascular endothelial cells were labeled. Basal membrane of the skin was identified by immunohistochemical anti-Laminin staining technique. Results  There was no obvious sign of acute rejection of the graft in both groups. The graft survival rate was 75.05%±3.69%, 83.12%±3.13% and 92.03%±3.87% at the 2th, 4th and 6th week respectively in the experimental group. The graft survival rate was 77.63%±3.23%, 83.17%±3.92% and 91.09%±3.35% at the 2th, 4th and 6th week in the control group. There was no significant difference between the two groups (P>0.05, but the contraction rate of the grafts was 9.13%±2.27%, 18.52%±3.40%, 23.92%±3.01% at the 2th, 4th, 6th week, respectively, in the experimental group, and 14.21%±3.05%, 29.12%±3

  12. Rapid detection of staphylococcal thermonuclease on casings of naturally contaminated fermented sausages.

    OpenAIRE

    Emswiler-Rose, B S; Johnston, R. W.; Harris, M E; Lee, W. H.

    1980-01-01

    Staphylococcal food poisoning associated with fermented sausages has been a recurring problem. By testing for thermonuclease by direct application of sausage casing disks on the surface of thermonuclease assay agar plates, possible Staphylococcus aureus growth in fermented sausages could be detected simply and rapidly. Koupal-Deibel deoxyribonucleic acid agar was somewhat superior to toluidine blue deoxyribonucleic acid agar for thermonuclease assay of fermented sausage casings. The sensitivi...

  13. Staphylococcal Enterotoxin A Detection from Rheumatoid Arthritis Patients’ Blood and Synovial Fluid

    OpenAIRE

    2016-01-01

    Introduction Direct detection of microbial super antigens in synovial fluid of patients with rheumatoid arthritis may be able to guide to the design of cost-effective therapies. The purpose of this study was to assess the existence of Staphylococcal enterotoxin A (superantigen A) in the synovial fluid of patients with RA by the PCR and ELISA methods. Methods This experimental study was conducted on the synovial fluid of 103 RA patients from Baqiyatallah University of Medical Sciences’ Rheumat...

  14. Antibody Production in Response to Staphylococcal MS-1 Phage Cocktail in Patients Undergoing Phage Therapy

    OpenAIRE

    Maciej Żaczek; Marzanna Łusiak-Szelachowska; Ewa Jończyk-Matysiak; Beata Weber-Dąbrowska; Ryszard Międzybrodzki; Barbara Owczarek; Agnieszka Kopciuch; Wojciech Fortuna; Paweł Rogóż; Andrzej Górski

    2016-01-01

    In this study, we investigated the humoral immune response (through the release of IgG, IgA, and IgM antiphage antibodies) to a staphylococcal phage cocktail in patients undergoing experimental phage therapy at the Phage Therapy Unit, Medical Center of the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy in Wrocław, Poland. We also evaluated whether occurring antiphage antibodies had neutralizing properties towards applied phages (K rate). Among 20 examined patients receiving...

  15. Chromatofocusing: a new method for purification of staphylococcal enterotoxins B and C1.

    OpenAIRE

    Ende, I A; Terplan, G; Kickhöfen, B.; Hammer, D K

    1983-01-01

    A new chromatographic procedure was developed which obtained highly purified preparations of staphylococcal enterotoxins B and C1 in yields of 60% from cultures of Staphylococcus aureus and which is faster than any of the separation methods used previously. The procedure involves chromatography on carboxymethylcellulose, removal of alpha-toxin by adsorption to rabbit erythrocyte membranes, and finally, chromatofocusing as the fundamental new step. Enterotoxins were obtained in highly purified...

  16. Strategies for Editing Virulent Staphylococcal Phages Using CRISPR-Cas10.

    Science.gov (United States)

    Bari, S M Nayeemul; Walker, Forrest C; Cater, Katie; Aslan, Barbaros; Hatoum-Aslan, Asma

    2017-09-21

    Staphylococci are prevalent skin-dwelling bacteria that are also leading causes of antibiotic-resistant infections. Viruses that infect and lyse these organisms (virulent staphylococcal phages) can be used as alternatives to conventional antibiotics and represent promising tools to eliminate or manipulate specific species in the microbiome. However, since over half their genes have unknown functions, virulent staphylococcal phages carry inherent risk to cause unknown downstream side effects. Further, their swift and destructive reproductive cycle make them intractable by current genetic engineering techniques. CRISPR-Cas10 is an elaborate prokaryotic immune system that employs small RNAs and a multisubunit protein complex to detect and destroy phages and other foreign nucleic acids. Some staphylococci naturally possess CRISPR-Cas10 systems, thus providing an attractive tool already installed in the host chromosome to harness for phage genome engineering. However, the efficiency of CRISPR-Cas10 immunity against virulent staphylococcal phages and corresponding utility as a tool to facilitate their genome editing has not been explored. Here, we show that the CRISPR-Cas10 system native to Staphylococcus epidermidis exhibits robust immunity against diverse virulent staphylococcal phages. On the basis of this activity, a general two-step approach was developed to edit these phages that relies upon homologous recombination machinery encoded in the host. Variations of this approach to edit toxic phage genes and access phages that infect CRISPR-less staphylococci are also presented. This versatile set of genetic tools enables the systematic study of phage genes of unknown functions and the design of genetically defined phage-based antimicrobials that can eliminate or manipulate specific Staphylococcus species.

  17. Outbreak of staphylococcal bullous impetigo in a maternity ward linked to an asymptomatic healthcare worker.

    Science.gov (United States)

    Occelli, P; Blanie, M; Sanchez, R; Vigier, D; Dauwalder, O; Darwiche, A; Provenzano, B; Dumartin, C; Parneix, P; Venier, A G

    2007-11-01

    An outbreak of staphylococcal bullous impetigo occurred over a period of five months in a maternity ward involving seven infected and two colonised neonates. The skin lesions were due to epidermolytic toxin A-producing Staphylococcus aureus. Infection control measures were implemented and a retrospective case-control study performed. Contact with an auxiliary nurse was the only risk factor for cases of bullous impetigo (Pimpetigo and the auxiliary nurse was reassigned to an adult ward.

  18. Potent Neutralization of Staphylococcal Enterotoxin B by Synergistic Action of Chimeric Antibodies▿

    OpenAIRE

    2010-01-01

    Staphylococcal enterotoxin B (SEB), a shock-inducing exotoxin synthesized by Staphylococcus aureus, is an important cause of food poisoning and is a class B bioterrorism agent. SEB mediates antigen-independent activation of a major subset of the T-cell population by cross-linking T-cell receptors (TCRs) with class II major histocompatibility complex (MHC-II) molecules of antigen-presenting cells, resulting in the induction of antigen independent proliferation and cytokine secretion by a signi...

  19. Marine Sponge-Derived Streptomyces sp. SBT343 Extract Inhibits Staphylococcal Biofilm Formation

    Science.gov (United States)

    Balasubramanian, Srikkanth; Othman, Eman M.; Kampik, Daniel; Stopper, Helga; Hentschel, Ute; Ziebuhr, Wilma; Oelschlaeger, Tobias A.; Abdelmohsen, Usama R.

    2017-01-01

    Staphylococcus epidermidis and Staphylococcus aureus are opportunistic pathogens that cause nosocomial and chronic biofilm-associated infections. Indwelling medical devices and contact lenses are ideal ecological niches for formation of staphylococcal biofilms. Bacteria within biofilms are known to display reduced susceptibilities to antimicrobials and are protected from the host immune system. High rates of acquired antibiotic resistances in staphylococci and other biofilm-forming bacteria further hamper treatment options and highlight the need for new anti-biofilm strategies. Here, we aimed to evaluate the potential of marine sponge-derived actinomycetes in inhibiting biofilm formation of several strains of S. epidermidis, S. aureus, and Pseudomonas aeruginosa. Results from in vitro biofilm-formation assays, as well as scanning electron and confocal microscopy, revealed that an organic extract derived from the marine sponge-associated bacterium Streptomyces sp. SBT343 significantly inhibited staphylococcal biofilm formation on polystyrene, glass and contact lens surfaces, without affecting bacterial growth. The extract also displayed similar antagonistic effects towards the biofilm formation of other S. epidermidis and S. aureus strains tested but had no inhibitory effects towards Pseudomonas biofilms. Interestingly the extract, at lower effective concentrations, did not exhibit cytotoxic effects on mouse fibroblast, macrophage and human corneal epithelial cell lines. Chemical analysis by High Resolution Fourier Transform Mass Spectrometry (HRMS) of the Streptomyces sp. SBT343 extract proportion revealed its chemical richness and complexity. Preliminary physico-chemical characterization of the extract highlighted the heat-stable and non-proteinaceous nature of the active component(s). The combined data suggest that the Streptomyces sp. SBT343 extract selectively inhibits staphylococcal biofilm formation without interfering with bacterial cell viability. Due to

  20. Methicillin-resistant Staphylococcus saprophyticus in Sweden carries various types of staphylococcal cassette chromosome mec (SCCmec).

    Science.gov (United States)

    Söderquist, B; Berglund, C

    2009-12-01

    Staphylococcus saprophyticus is a common cause of uncomplicated urinary tract infections and is usually susceptible to the antimicrobial agents used for their treatment. However, S. saprophyticus resistant to beta-lactam antibiotics and carrying mecA has been reported. Eight Swedish isolates of mecA-positive S. saprophyticus with diverse origin carrying at least three different types of staphylococcal cassette chromosome mec (SCCmec) are described here.

  1. Rapid detection of staphylococcal thermonuclease on casings of naturally contaminated fermented sausages.

    OpenAIRE

    Emswiler-Rose, B S; Johnston, R. W.; Harris, M E; Lee, W. H.

    1980-01-01

    Staphylococcal food poisoning associated with fermented sausages has been a recurring problem. By testing for thermonuclease by direct application of sausage casing disks on the surface of thermonuclease assay agar plates, possible Staphylococcus aureus growth in fermented sausages could be detected simply and rapidly. Koupal-Deibel deoxyribonucleic acid agar was somewhat superior to toluidine blue deoxyribonucleic acid agar for thermonuclease assay of fermented sausage casings. The sensitivi...

  2. Staphylococcal enterotoxin H induced apoptosis of bovine mammary epithelial cells in vitro.

    Science.gov (United States)

    Liu, Yongxia; Chen, Wei; Ali, Tariq; Alkasir, Rashad; Yin, Jinhua; Liu, Gang; Han, Bo

    2014-12-19

    Staphylococcal enterotoxins (SEs) are powerful superantigenic toxins produced by Staphylococcus aureus (S. aureus). They can cause food poisoning and toxic shock. However, their impact on bovine mammary epithelial cells (bMECs) is still unknown. In this study, the distribution of SE genes was evaluated in 116 S. aureus isolates from bovine mastitis, and the most prevalent genes were seh (36.2%), followed by sei (12.1%), seg (11.2%), ser (4.3%), sec (3.4%), sea (2.6%) and sed (1.7%). To better understand the effect of staphylococcal enterotoxin H (SEH) on bMECs, the seh gene was cloned and inserted into the prokaryotic expression vector, pET28a, and transformed into Escherichia coli BL21 (DE3). The recombinant staphylococcal enterotoxin H (rSEH) was expressed and purified as soluble protein. Bioactivity analysis showed that rSEH possessed the activity of stimulating lymphocytes proliferation. The XTT assay showed that 100 μg/mL of rSEH produced the cytotoxic effect on bMECs, and fluorescence microscopy and flow cytometry analysis revealed that a certain dose of rSEH is effective at inducing bMECs apoptosis in vitro. This indicates that SEs can directly lead to cellular apoptosis of bMECs in bovine mastitis associated with S. aureus.

  3. xMAP-based analysis of three most prevalent staphylococcal toxins in Staphylococcus aureus cultures.

    Science.gov (United States)

    Simonova, Maria A; Petrova, Elena E; Dmitrenko, Olga A; Komaleva, Ravilya L; Shoshina, Natalia S; Samokhvalova, Larisa V; Valyakina, Tatiana I; Grishin, Eugene V

    2014-10-01

    Detection of staphylococcal toxins presents a great interest for medical diagnostics. Screening of clinical samples for the presence of several types of staphylococcal toxins using traditional methods-biological tests on animals or cell cultures as well as ELISA-is laborious. Multiplex detection methods would simplify testing. We have designed an xMAP-based assay to detect three staphylococcal toxins-enterotoxins A and B (SEA and SEB) and toxic shock syndrome toxin (TSST)-in cultural supernatants obtained from different strains of Staphylococcus aureus. The limits of detection of SEA, SEB, and TSST multiplex detection in S. aureus growth medium were 10, 1,000, and 5 pg/mL, respectively. Fifty-nine samples of S. aureus cultural supernatants were tested with the xMAP assay. The developed assay has proved highly effective detection of the natural toxins in the samples obtained due to bacterial cells cultivation. In prospect, the developed test system can be used in clinical diagnostics and in monitoring of foodstuffs and environmental objects.

  4. Molecular Typing of Staphylococcus Aureus Isolate Responsible for Staphylococcal Poisoning Incident in Homemade Food

    Science.gov (United States)

    Bellio, Alberto; Bianchi, Daniela Manila; Gallina, Silvia; Adriano, Daniela; Zuccon, Fabio; Chiesa, Francesco; Acutis, Pier Luigi; Casalinuovo, Francesco; Decastelli, Lucia

    2016-01-01

    In October 2012, two persons fell ill with symptoms consistent with staphylococcal food poisoning after eating home-canned tuna fish and tomatoes. Laboratory investigation detected the enterotoxins in the home-canned tuna and molecular analysis of the isolated Staphylococcus aureus confirmed it carried toxin genes. Qualitative enzyme-linked immunosorbent assay and enzime linked fluorescent assay methods and quantitative assay identified the enterotoxins in the food leftovers, specifically staphylococcal enterotoxins type A (SEA) and D (SED), respectively 0.49 and 2.04 ng/g. The laboratory results are discussed considering the relation to the fish in oil, survival and heat resistance of S. aureus, and presumptive microbial contamination due to improper handling during home-canning procedures. This is the first reported cluster of foodborne illnesses due to staphylococcal enterotoxins in tuna in Italy. In this study, we reported cases described and analysed for their spa-type. Showing a high heterogeneity of isolates, spa-type t13252 is correlated in a node of the minimum spanning tree and it has never been reported as responsible for foodborne outbreak. This case underlines the importance of risk communication and dissemination of home-canning guidelines to reduce the incidence of foodborne outbreaks caused by homemade conserves.

  5. Impact of Environmental Cues on Staphylococcal Quorum Sensing and Biofilm Development.

    Science.gov (United States)

    Kavanaugh, Jeffrey S; Horswill, Alexander R

    2016-06-10

    Staphylococci are commensal bacteria that colonize the epithelial surfaces of humans and many other mammals. These bacteria can also attach to implanted medical devices and develop surface-associated biofilm communities that resist clearance by host defenses and available chemotherapies. These communities are often associated with persistent staphylococcal infections that place a tremendous burden on the healthcare system. Understanding the regulatory program that controls staphylococcal biofilm development, as well as the environmental conditions that modulate this program, has been a focal point of research in recent years. A central regulator controlling biofilm development is a peptide quorum-sensing system, also called the accessory gene regulator or agr system. In the opportunistic pathogen Staphylococcus aureus, the agr system controls production of exo-toxins and exo-enzymes essential for causing infections, and simultaneously, it modulates the ability of this pathogen to attach to surfaces and develop a biofilm, or to disperse from the biofilm state. In this review, we explore advances on the interconnections between the agr quorum-sensing system and biofilm mechanisms, and topics covered include recent findings on how different environmental conditions influence quorum sensing, the impact on biofilm development, and ongoing questions and challenges in the field. As our understanding of the quorum sensing and biofilm interconnection advances, there are growing opportunities to take advantage of this knowledge and develop therapeutic approaches to control staphylococcal infections.

  6. Meta-analysis of staphylococcal diarrhea in some developing African countries

    Directory of Open Access Journals (Sweden)

    George P. Einstein

    2017-06-01

    Full Text Available Staphylococcal diarrhea is a common gastro-intestinal illness caused by poor water supply and unhygienic food preparation. Although, about 70% of diarrheal cases per year have been attributed to the consumption of contaminated foods, in Africa, few studies have reported the incidence of Staphylococcus-related diarrhea and the implicated virulence factors. We investigated the implications of common factors such as age of children, age of mothers, maternal level of education, gender of children, overall bacteria isolated, sanitary condition status and feeding type of mothers on the burden of staphylococcal diarrhea. A literature search was performed using Pubmed, Googlescholar, American society for microbiology (ASM journals and other sources. The quality of studies was assessed. Adjusted odds ratios with 95% confidence intervals (CI for the occurrence of Staphylococcus aureus amongst other causes were extracted. For each study, a multivariate logistic regression analysis of the adjusted odds ratio was performed to identify the risk factors on the burden of staphylococcal diarrhea. Six case-control studies were included in the meta–analysis. Quality of individual studies rages from 0.57–0.84 (median, 0.69. meta-analysis gave pooled odds ratios 3.27 (95% CI; 2.88–4.17. The results identified inadequate sanitary conditions and type of feeding as major risk factors.

  7. Molecular Typing of Staphylococcus Aureus Isolate Responsible for Staphylococcal Poisoning Incident in Homemade Food.

    Science.gov (United States)

    Macori, Guerrino; Bellio, Alberto; Bianchi, Daniela Manila; Gallina, Silvia; Adriano, Daniela; Zuccon, Fabio; Chiesa, Francesco; Acutis, Pier Luigi; Casalinuovo, Francesco; Decastelli, Lucia

    2016-04-19

    In October 2012, two persons fell ill with symptoms consistent with staphylococcal food poisoning after eating home-canned tuna fish and tomatoes. Laboratory investigation detected the enterotoxins in the home-canned tuna and molecular analysis of the isolated Staphylococcus aureus confirmed it carried toxin genes. Qualitative enzyme-linked immunosorbent assay and enzime linked fluorescent assay methods and quantitative assay identified the enterotoxins in the food leftovers, specifically staphylococcal enterotoxins type A (SEA) and D (SED), respectively 0.49 and 2.04 ng/g. The laboratory results are discussed considering the relation to the fish in oil, survival and heat resistance of S. aureus, and presumptive microbial contamination due to improper handling during home-canning procedures. This is the first reported cluster of foodborne illnesses due to staphylococcal enterotoxins in tuna in Italy. In this study, we reported cases described and analysed for their spa-type. Showing a high heterogeneity of isolates, spa-type t13252 is correlated in a node of the minimum spanning tree and it has never been reported as responsible for foodborne outbreak. This case underlines the importance of risk communication and dissemination of home-canning guidelines to reduce the incidence of foodborne outbreaks caused by homemade conserves.

  8. Molecular typing of Staphylococcus aureus isolate responsible for staphylococcal poisoning incident in homemade food

    Directory of Open Access Journals (Sweden)

    Guerrino Macori

    2016-06-01

    Full Text Available In October 2012, two persons fell ill with symptoms consistent with staphylococcal food poisoning after eating home-canned tuna fish and tomatoes. Laboratory investigation detected the enterotoxins in the home-canned tuna and molecular analysis of the isolated Staphylococcus aureus confirmed it carried toxin genes. Qualitative enzyme-linked immunosorbent assay and enzime linked fluorescent assay methods and quantitative assay identified the enterotoxins in the food leftovers, specifically staphylococcal enterotoxins type A (SEA and D (SED, respectively 0.49 and 2.04 ng/g. The laboratory results are discussed considering the relation to the fish in oil, survival and heat resistance of S. aureus, and presumptive microbial contamination due to improper handling during home-canning procedures. This is the first reported cluster of foodborne illnesses due to staphylococcal enterotoxins in tuna in Italy. In this study, we reported cases described and analysed for their spa-type. Showing a high heterogeneity of isolates, spa-type t13252 is correlated in a node of the minimum spanning tree and it has never been reported as responsible for foodborne outbreak. This case underlines the importance of risk communication and dissemination of home-canning guidelines to reduce the incidence of foodborne outbreaks caused by homemade conserves.

  9. Human Monocyte-Derived Osteoclasts Are Targeted by Staphylococcal Pore-Forming Toxins and Superantigens.

    Directory of Open Access Journals (Sweden)

    Sacha Flammier

    Full Text Available Staphylococcus aureus is the leading cause of bone and joint infections (BJIs. Staphylococcal pathogenesis involves numerous virulence factors including secreted toxins such as pore-forming toxins (PFTs and superantigens. The role of these toxins on BJI outcome is largely unknown. In particular, few studies have examined how osteoclasts, the bone-resorbing cells, respond to exposure to staphylococcal PFTs and superantigens. We investigated the direct impact of recombinant staphylococcal toxins on human primary mature monocyte-derived osteoclasts, in terms of cytotoxicity and cell activation with cell death and bone resorption assays, using macrophages of the corresponding donors as a reference. Monocyte-derived osteoclasts displayed similar toxin susceptibility profiles compared to macrophages. Specifically, we demonstrated that the Panton-Valentine leukocidin, known as one of the most powerful PFT which lyses myeloid cells after binding to the C5a receptor, was able to induce the death of osteoclasts. The archetypal superantigen TSST-1 was not cytotoxic but enhanced the bone resorption activity of osteoclasts, suggesting a novel mechanism by which superantigen-producing S. aureus can accelerate the destruction of bone tissue during BJI. Altogether, our data indicate that the diverse clinical presentations of BJIs could be related, at least partly, to the toxin profiles of S. aureus isolates involved in these severe infections.

  10. Effective removal of staphylococcal biofilms by the endolysin LysH5.

    Directory of Open Access Journals (Sweden)

    Diana Gutiérrez

    Full Text Available Staphylococcal biofilms are a major concern in both clinical and food settings because they are an important source of contamination. The efficacy of established cleaning procedures is often hindered due to the ability of some antimicrobial compounds to induce biofilm formation, and to the presence of persister cells, a small bacterial subpopulation that exhibits multidrug tolerance. Phage lytic enzymes have demonstrated antimicrobial activity against planktonic and sessile bacteria. However, their ability to lyse and/or select persister cells remains largely unexplored so far. In this work, the lytic activity of the endolysin LysH5 against Staphylococcus aureus and Staphylococcus epidermidis biofilms was confirmed. LysH5 reduced staphylococcal sessile cell counts by 1-3 log units, compared with the untreated control, and sub-inhibitory concentrations of this protein did not induce biofilm formation. LysH5-surviving cells were not resistant to the lytic activity of this protein, suggesting that no persister cells were selected. Moreover, to prove the lytic ability of LysH5 against this subpopulation, both S. aureus exponential cultures and persister cells obtained after treatment with rifampicin and ciprofloxacin were subsequently treated with LysH5. The results demonstrated that besides the notable activity of endolysin LysH5 against staphylococcal biofilms, persister cells were also inhibited, which raises new opportunities as an adjuvant for some antibiotics.

  11. Effective removal of staphylococcal biofilms by the endolysin LysH5.

    Science.gov (United States)

    Gutiérrez, Diana; Ruas-Madiedo, Patricia; Martínez, Beatriz; Rodríguez, Ana; García, Pilar

    2014-01-01

    Staphylococcal biofilms are a major concern in both clinical and food settings because they are an important source of contamination. The efficacy of established cleaning procedures is often hindered due to the ability of some antimicrobial compounds to induce biofilm formation, and to the presence of persister cells, a small bacterial subpopulation that exhibits multidrug tolerance. Phage lytic enzymes have demonstrated antimicrobial activity against planktonic and sessile bacteria. However, their ability to lyse and/or select persister cells remains largely unexplored so far. In this work, the lytic activity of the endolysin LysH5 against Staphylococcus aureus and Staphylococcus epidermidis biofilms was confirmed. LysH5 reduced staphylococcal sessile cell counts by 1-3 log units, compared with the untreated control, and sub-inhibitory concentrations of this protein did not induce biofilm formation. LysH5-surviving cells were not resistant to the lytic activity of this protein, suggesting that no persister cells were selected. Moreover, to prove the lytic ability of LysH5 against this subpopulation, both S. aureus exponential cultures and persister cells obtained after treatment with rifampicin and ciprofloxacin were subsequently treated with LysH5. The results demonstrated that besides the notable activity of endolysin LysH5 against staphylococcal biofilms, persister cells were also inhibited, which raises new opportunities as an adjuvant for some antibiotics.

  12. Lactobacilli require physical contact to reduce staphylococcal TSST-1 secretion and vaginal epithelial inflammatory response.

    Science.gov (United States)

    Younes, Jessica A; Reid, Gregor; van der Mei, Henny C; Busscher, Henk J

    2016-06-01

    ITALIC! Staphylococcus aureusbiofilms can be found on vaginal epithelia, secreting toxins and causing inflammation. The co-vaginal species ITALIC! Lactobacilluscan alter staphylococcal-induced epithelial secretion of inflammatory cytokines and quench staphylococcal toxic shock syndrome toxin-1 secretion. It is hypothesized that these effects of lactobacilli require direct physical contact between lactobacilli, staphylococci and the epithelium. Indeed, lactobacilli only reduced ITALIC! S. aureus-induced inflammatory cytokine expression when allowed physical contact with vaginal epithelial cells. Furthermore, a reduction in toxic shock syndrome toxin-1 secretion only occurred when a probiotic ITALIC! Lactobacillusstrain was allowed contact, but not when being physically separated from ITALIC! S. aureus Bacterial-probe atomic force microscopy demonstrated that lactobacilli and staphylococci strongly adhere to epithelial cells, while lactobacilli adhere stronger to staphylococci than staphylococci to each other, giving lactobacilli opportunity to penetrate and reside in staphylococcal biofilms, as visualized using confocal laser scanning microscopy with fluorescence ITALIC! in situhybridization probes. These results identify that physical contact and biochemical signaling by lactobacilli are intrinsically linked mechanisms that reduce virulence of ITALIC! S. aureusbiofilm.

  13. Population-based resequencing analysis of wild and cultivated barley revealed weak domestication signal of selection and bottleneck in the Rrs2 scald resistance gene region.

    Science.gov (United States)

    Fu, Yong-Bi

    2012-02-01

    Many plant disease resistance (R) genes have been cloned, but the potential of utilizing these plant R-gene genomic resources for genetic inferences of plant domestication history remains unexplored. A population-based resequencing analysis of the genomic region near the Rrs2 scald resistance gene was made in 51 accessions of wild and cultivated barley from 41 countries. Fifteen primer pairs were designed to sample the genomic region with a total length of 10 406 bp. More nucleotide diversity was found in wild (π = 0.01846) than cultivated (π = 0.01507) barley samples. Three distinct groups of 29 haplotypes were detected for all 51 samples, and they were well mixed with wild and cultivated barley samples from different countries and regions. The neutrality tests by Tajima's D were not significant, but a significant (P domestication. Together, the domestication signal in the genomic region was weak both in human selection and domestication bottleneck.

  14. PCR primers for the detection of staphylococcal enterotoxins K, L, and M and survey of staphylococcal enterotoxin types in Staphylococcus aureus isolates from food poisoning cases in Taiwan.

    Science.gov (United States)

    Chiang, Yu-Cheng; Chang, Li-Tung; Lin, Chia-Wei; Yang, Chi-Yea; Tsen, Hau-Yang

    2006-05-01

    Staphylococcal enterotoxins (SEs) are important causative agents in gastroenteritidis and food poisoning cases. They are serologically grouped into five major classical types, i.e., SEA, SEB, SEC, SED, and SEE. In addition, new SEs, such as SEG through SEM, have recently been identified and characterized. In an attempt to survey the distribution of classical and new SEs in organisms responsible for staphylococcal infections in Taiwan, we developed PCR primers for the genes that define the SEK, SEL, and SEM types. Bacterial strains other than sek, sel, and sem Staphylococcus aureus, including strains of other Staphylococcus species, did not generate any false-positive results when examined with these primers. The expression potential for the sek, sel, and sem types were also determined by reverse transcription-PCR. Together with the PCR primers specific for the classical SEs and other new SEs, including SEG, SEH, SEI, and SEJ, we surveyed the SE genes in S. aureus strains isolated from food poisoning cases. For 147 S. aureus isolates originating from food poisoning cases, 109 (74.1%) were positive for one or more SE genes. Of them, the major classical enterotoxin type was sea (28.6%), followed by seb (20.4%), sec (8.2%), and sed (2.0%). For the new SE types, sei (30.6%) was detected the most often, followed by sek (18.4%), sem (12.9%), and sel (8.2%). Also, 64 (43.5%) of the total bacterial strains had more than one enterotoxin gene.

  15. Morphological changes of Endothelin-1 innervation in cerebral arteries of scald rats%烫伤大鼠脑底动脉壁内含内皮素-1能神经纤维的形态学变化

    Institute of Scientific and Technical Information of China (English)

    林雪群; 祝高春; 薛国勇; 唐讯

    2012-01-01

    目的 探讨烫(烧)伤损伤时大鼠脑血管内皮素-1能神经纤维分布与脑血管神经源性调节的关系,以及烫(烧)伤对脑血管损伤的影响.方法应用免疫组织化学技术观察烫(烧)伤大鼠脑底动脉(包括大脑前动脉、大脑中动脉、大脑后动脉和基底动脉)内皮素-1能神经纤维的分布.结果 烫(烧)伤大鼠和正常大鼠脑底动脉均可见棕褐色的内皮素-1能免疫反应阳性神经纤维,似细线状,攀附于血管壁上,烫(烧)伤大鼠脑底动脉各主要分支内皮素-1能免疫反应阳性纤维密度较正常大鼠明显增加,纤维走行大多呈网状.结论 烫(烧)伤可引起大鼠脑底动脉内皮素-1能免疫反应阳性神经纤维增加,增加的内皮素-1能神经纤维可能诱发脑血管痉挛和脑血液循环紊乱.提示内皮素-1能神经纤维在烫(烧)伤后在脑血管的神经源性调节中可能起重要的作用.%Objective To investigate the relationship between ET - 1 innervation and neurogenic regulation of cerebral vessels during scald, and to detect the effect of scald-induced cerebrovascular injury. Methods Immunohistochemical technique was used to measure the density of endothelin-1 nerve fibers in cerebral arteries of scald rat. Results The density of ET - 1 nerve fibers in cerebral arteries of scald rats was obviously higher than that of the control group. Conclusion Scald may cause an increase in the density of ET-1 nerve fibers in cerebral arteries. The elevated ET - 1 innervation may induce cerebral vasospasm and promote the disorder of cerebral blood circulation. It is suggested that ET-1 innervation may play an important role in the neurogenic regulation of cerebral vessels after scald.

  16. Variation in the genomic locations and sequence conservation of STAR elements among staphylococcal species provides insight into DNA repeat evolution

    Directory of Open Access Journals (Sweden)

    Purves Joanne

    2012-09-01

    Full Text Available Abstract Background Staphylococcus aureus Repeat (STAR elements are a type of interspersed intergenic direct repeat. In this study the conservation and variation in these elements was explored by bioinformatic analyses of published staphylococcal genome sequences and through sequencing of specific STAR element loci from a large set of S. aureus isolates. Results Using bioinformatic analyses, we found that the STAR elements were located in different genomic loci within each staphylococcal species. There was no correlation between the number of STAR elements in each genome and the evolutionary relatedness of staphylococcal species, however higher levels of repeats were observed in both S. aureus and S. lugdunensis compared to other staphylococcal species. Unexpectedly, sequencing of the internal spacer sequences of individual repeat elements from multiple isolates showed conservation at the sequence level within deep evolutionary lineages of S. aureus. Whilst individual STAR element loci were demonstrated to expand and contract, the sequences associated with each locus were stable and distinct from one another. Conclusions The high degree of lineage and locus-specific conservation of these intergenic repeat regions suggests that STAR elements are maintained due to selective or molecular forces with some of these elements having an important role in cell physiology. The high prevalence in two of the more virulent staphylococcal species is indicative of a potential role for STAR elements in pathogenesis.

  17. Variation in the genomic locations and sequence conservation of STAR elements among staphylococcal species provides insight into DNA repeat evolution.

    Science.gov (United States)

    Purves, Joanne; Blades, Matthew; Arafat, Yasrab; Malik, Salman A; Bayliss, Christopher D; Morrissey, Julie A

    2012-09-28

    Staphylococcus aureus Repeat (STAR) elements are a type of interspersed intergenic direct repeat. In this study the conservation and variation in these elements was explored by bioinformatic analyses of published staphylococcal genome sequences and through sequencing of specific STAR element loci from a large set of S. aureus isolates. Using bioinformatic analyses, we found that the STAR elements were located in different genomic loci within each staphylococcal species. There was no correlation between the number of STAR elements in each genome and the evolutionary relatedness of staphylococcal species, however higher levels of repeats were observed in both S. aureus and S. lugdunensis compared to other staphylococcal species. Unexpectedly, sequencing of the internal spacer sequences of individual repeat elements from multiple isolates showed conservation at the sequence level within deep evolutionary lineages of S. aureus. Whilst individual STAR element loci were demonstrated to expand and contract, the sequences associated with each locus were stable and distinct from one another. The high degree of lineage and locus-specific conservation of these intergenic repeat regions suggests that STAR elements are maintained due to selective or molecular forces with some of these elements having an important role in cell physiology. The high prevalence in two of the more virulent staphylococcal species is indicative of a potential role for STAR elements in pathogenesis.

  18. APPLICATION OF THE HACCP METHOD IN THE SWINE ABATTOIR BY SCALDING-SKINNING ECOLOGICAL PROCESSING / APPLICATION DE LA METHODE HACCP DANS L’ABATTOIR PORCIN PAR LE PROCESSUS ECOLOGIQUE D’ÉCHAUDAGE-DÉPEÇAGE

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    IULIAN SUCEVEANU

    2017-03-01

    Full Text Available In the food industry development, the food safety and the valorization of by-products represent important aspects. The present study consists in the application of the Hazard Analysis and Critical Control Point (HACCP method in pig slaughterhouses for scalding-skinning technology in order to ensure the conformity of meat quality and the by-products management and recycling. In order to optimize the scalding-skinning process of swine, all the classical technological steps of the technological processes were analyzed separately and globally, with the adaptation of the HACCP method for the new technology. The result of this study shows that the implementation of the new technology that combines existing technologies has many advantages from both an economic and a technological point of view. At the same time the new technology ensures food safety by taking account of the application of the food safety system, respectively, HACCP method.

  19. The clinical spectrum and antibiotic sensitivity patterns of staphylococcal pyodermas in the community and hospital

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    Shireen Furtado

    2014-01-01

    Full Text Available Context: The uncontrolled use of antibiotics has resulted in a relentless spread of multiresistant strains of Staphylococcus aureus. There are studies conducted in medical colleges in Chandigarh, Chennai, Mumbai and Vellore comparing pyodermas in the community and hospital setting based on clinical and bacteriological parameters. Aims: This study, conducted over 1½ years from March 2009 to August 2010, aimed at analyzing the clinical spectrum and antibiotic sensitivity pattern of community and hospital-associated (HA staphylococcal pyoderma. It also assessed the prevalence of methicillin-resistant S. aureus (MRSA in the community and hospital cohort settings. Subjects and Methods: The study comprised of 200 cases of staphylococcal pyodermas, derived from the community (150 cases and hospital (50 cases. Patients were evaluated based on their clinical presentation; antibiotic susceptibility was tested using the Kirby-Bauer disk diffusion method. Statistical Analysis Used: Statistical significance between individual attributes between the community and HA staphylococcal pyoderma groups was analyzed using Chi-square test and mean differences using student′s t-test. Results: Factors associated with community-associated (CA pyodermas were young age (P = 0.0021, primary pyodermas, and involvement of extremities, while those with HA pyodermas were middle age, secondary pyodermas, and significantly increased body surface involvement (P = 0.041. Incidence of CA-MRSA was 11.3%, while that of HA-MRSA was 18%. Conclusions: A high level of resistance to first-line drugs such as penicillin, ciprofloxacin and cotrimoxazole was observed, more so in the hospital strain than in the community strain. S. aureus demonstrated good susceptibility to cephalosporins. Though the two strains of MRSA differed clinically, they showed 100% sensitivity to vancomycin and linezolid.

  20. Staphylococcal Enterotoxin A Detection from Rheumatoid Arthritis Patients’ Blood and Synovial Fluid

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    Ataee, Ramezan Ali; Kahani, Mahboobeh Sadat; Alishiri, Gholam Hossein; Ahamadi, Zyenab

    2016-01-01

    Introduction Direct detection of microbial super antigens in synovial fluid of patients with rheumatoid arthritis may be able to guide to the design of cost-effective therapies. The purpose of this study was to assess the existence of Staphylococcal enterotoxin A (superantigen A) in the synovial fluid of patients with RA by the PCR and ELISA methods. Methods This experimental study was conducted on the synovial fluid of 103 RA patients from Baqiyatallah University of Medical Sciences’ Rheumatology Clinic in Tehran, Iran in 2011–2014. Bacterial cultures, polymerase chain reaction with specific primer pairs and enzyme-linked immunosorbent assay (ELISA) methods were used. The PCR products were subjected to sequence as a confirmatory molecular method results. The data were descriptively analyzed by SPSS Version 19. Results The bacteriological study result indicated that, in four cases (3.8%) of the patients, bacterial strains were isolated. The result of PCR molecular method for staphylococcal enterotoxin A gene showed that, 42 of the patients (40.7%) tested positive for the ent A gene. The results of ELISA were positive for staphylococcal enterotoxin A (superantigen A) in 51 cases (49.51%) of the patients’ synovial fluids. The results indicated that the possibility of detecting superantigen A in the SF of RA patients, but the origin of the enterotoxin A gene remained unknown. Conclusions The findings of this study may be able to alter the actual theory on the pathogenesis, diagnosis, and treatment of RA patients. In addition, the results have shown the probability of an endogenous origin for the involved superantigen A in RA patients’ synovial fluids. PMID:27053990

  1. Staphylococcus aureus isolates encode variant staphylococcal enterotoxin B proteins that are diverse in superantigenicity and lethality.

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    Petra L Kohler

    Full Text Available Staphylococcus aureus produces superantigens (SAgs that bind and cross-link T cells and APCs, leading to activation and proliferation of immune cells. SAgs bind to variable regions of the β-chains of T cell receptors (Vβ-TCRs, and each SAg binds a unique subset of Vβ-TCRs. This binding leads to massive cytokine production and can result in toxic shock syndrome (TSS. The most abundantly produced staphylococcal SAgs and the most common causes of staphylococcal TSS are TSS toxin-1 (TSST-1, and staphylococcal enterotoxins B and C (SEB and SEC, respectively. There are several characterized variants of humans SECs, designated SEC1-4, but only one variant of SEB has been described. Sequencing the seb genes from over 20 S. aureus isolates show there are at least five different alleles of seb, encoding forms of SEB with predicted amino acid substitutions outside of the predicted immune-cell binding regions of the SAgs. Examination of purified, variant SEBs indicates that these amino acid substitutions cause differences in proliferation of rabbit splenocytes in vitro. Additionally, the SEBs varied in lethality in a rabbit model of TSS. The SEBs were diverse in their abilities to cause proliferation of human peripheral blood mononuclear cells, and differed in their activation of subsets of T cells. A soluble, high-affinity Vβ-TCR, designed to neutralize the previously characterized variant of SEB (SEB1, was able to neutralize the variant SEBs, indicating that this high-affinity peptide may be useful in treating a variety of SEB-mediated illnesses.

  2. Plant plasma membrane-bound staphylococcal-like DNases as a novel class of eukaryotic nucleases

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    Leśniewicz Krzysztof

    2012-10-01

    Full Text Available Abstract Background The activity of degradative nucleases responsible for genomic DNA digestion has been observed in all kingdoms of life. It is believed that the main function of DNA degradation occurring during plant programmed cell death is redistribution of nucleic acid derived products such as nitrogen, phosphorus and nucleotide bases. Plant degradative nucleases that have been studied so far belong mainly to the S1-type family and were identified in cellular compartments containing nucleic acids or in the organelles where they are stored before final application. However, the explanation of how degraded DNA components are exported from the dying cells for further reutilization remains open. Results Bioinformatic and experimental data presented in this paper indicate that two Arabidopsis staphylococcal-like nucleases, named CAN1 and CAN2, are anchored to the cell membrane via N-terminal myristoylation and palmitoylation modifications. Both proteins possess a unique hybrid structure in their catalytic domain consisting of staphylococcal nuclease-like and tRNA synthetase anticodon binding-like motifs. They are neutral, Ca2+-dependent nucleaces showing a different specificity toward the ssDNA, dsDNA and RNA substrates. A study of microarray experiments and endogenous nuclease activity revealed that expression of CAN1 gene correlates with different forms of programmed cell death, while the CAN2 gene is constitutively expressed. Conclusions In this paper we present evidence showing that two plant staphylococcal-like nucleases belong to a new, as yet unidentified class of eukaryotic nucleases, characterized by unique plasma membrane localization. The identification of this class of nucleases indicates that plant cells possess additional, so far uncharacterized, mechanisms responsible for DNA and RNA degradation. The potential functions of these nucleases in relation to their unique intracellular location are discussed.

  3. Staphylococcal Cassette Chromosome mec Types Among Methicillin-Resistant Staphylococcus aureus in Northern Iran

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    Taherirad, Akram; Jahanbakhsh, Roghayeh; Shakeri, Fatemeh; Anvary, Shaghayegh; Ghaemi, Ezzat Allah

    2016-01-01

    Background Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of nosocomial and community-acquired infections around the world. Staphylococcal cassette chromosome mec (SCCmec) typing methods are often used to study MRSA molecular epidemiology. Objectives The current study was designed to explore the distribution profiles of different SCCmec types among methicillin-resistant S. aureus strains isolated from hospitals in Gorgan, in northern Iran, and to correlate the types into observed bacterial virulence factors. Materials and Methods Staphylococcal cassette chromosome mec typing of 62 MRSA strains isolated from patients and health-care workers in Gorgan was performed using multiplex polymerase chain reaction (PCR) assay. The prevalence of the strains was then compared according to isolation source, antibiotic susceptibility profiles, biofilm production, and the presence of the Panton-Valentine gene in isolates. Results The most common SCCmec type was type III, with a frequency rate of 76%, followed by types IV, I, and V, with frequency rates of 11.2%, 4.8%, and 3.2%, respectively; three isolates (4.8%) were not typeable by this method. SCCmec type I was only isolated from blood culture, and types IV and V were mainly isolated from wounds and urine samples; SCCmec type III was isolated from all of the clinically samples. All of the MRSA strains that were isolated from healthy carriers were type III. Multidrug resistance in the type III strains was higher compared to the other types. The frequencies of Panton-Valentine and biofilm production were significantly lower in the type III strains compared to the other SCCmec types (P < 0.05). Conclusions Similarly to other geographical regions of Iran, the SCCmec type III MRSA strain was the most frequently isolated strain from patients in Gorgan. Staphylococcal cassette chromosome mec type III showed fewer virulence factors compared to other SCCmec types. PMID:27800133

  4. Staphylococcal superantigens; toxic shock syndrome toxin-1 and enterotoxins in pediatric otitis media effusion: a brief report

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    Noorbakhsh S

    2013-03-01

    Full Text Available Background: Staphylococcal superantigens (SAg's may have some role in otitis media with effusion (OME. The aim of this study was the search of staphylococcal SAg's in middle ear effusion of children with OME.  Methods: This cross sectional-analytic study was done in ENT & pediatric wards upon 64 children with otitis media with effusion (OME between 1-15 years, (mean age=7.42+4 years of Rasoul Akram University Hospital, Tehran, Iran in 2009-2011. Fifty six percent (36 of cases were male, 43.8% (28 were female. Staphylococcal SAg's; Toxic Shock Syndrome Toxin-1 (TSST-1, Staphylococcal enterotoxin A, B, C, D (Enzyme immune assay, AB Cam, USA were detected in middle ear effusion samples after conventional culture.Results: None type of SAg's found in 39% of OME cases, enterotoxin B found in: 22%; enterotoxin A: 17%, enterotoxin C: 15.6%, enterotoxin D: 12.5%, Toxic Shock Syndrome Toxin-1 (TSST-1: 7.8% Mean age of cases with positive TSST-1, enterotoxin A, B, C, and D was: 1, 5, 8.6, 9.6 and 9.6 years respectively. Positive TSST had no agreement with positive enterotoxin A and C but had weak agreement with type B and D. Mean age of cases with positive TSST was one years which had significant difference with (7.9 years in cases with negative TSST test (P<0.0001.Conclusion: At least one or more type of staphylococcal toxins had found in middle ear effusion of 70% of OME cases with negative culture for Staphylococcus aureus. Even in culture negative cases, staphylococcal toxins might have some immunologic role in middle ear effusion forming. Finding the SAg's (at least one type are important for treatment of immunosuppressive or corticosteroid in cases with resistant OME.

  5. Ultrafast solvation dynamics at internal site of staphylococcal nuclease investigated by site-directed mutagenesis

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    Guang-yu, Gao; Wei, Wang; Shu-feng, Wang; Zhong, Dongping; Qi-huang, Gong

    2014-01-01

    Solvation is essential for protein activities. To study internal solvation of protein, site-directed mutagenesis is applied. Intrinsic fluorescent probe, tryptophan, is inserted into desired position inside protein molecule for ultrafast spectroscopic study. Here we review this unique method for protein dynamics researches. We introduce the frontiers of protein solvation, site-directed mutagenesis, protein stability and characteristics, and the spectroscopic methods. Then we present time-resolved spectroscopic dynamics of solvation dynamics inside caves of active sites. The studies are carried out on a globular protein, staphylococcal nuclease. The solvation at internal sites of the caves indicate clear characteristics of local environment. These solvation behaviors correlated to the enzyme activity directly.

  6. Prevalence of staphylococcal enterotoxins in Staphylococcus pseudintermedius isolates from dogs with pyoderma and healthy dogs.

    Science.gov (United States)

    Tanabe, Taishi; Toyoguchi, Midori; Hirano, Fumitaka; Chiba, Mei; Onuma, Kenta; Sato, Hisaaki

    2013-09-01

    To investigate the role of staphylococcal enterotoxins (SEs) produced by Staphylococcus pseudintermedius in the pathogenesis of pyoderma, isolates from dogs with pyoderma and healthy dogs were analyzed. According to reverse passive latex agglutination, 14/184 isolates (7.6%) from dogs with pyoderma and 9/87 (10.3%) from healthy dogs produced SEs (SEA, SEC or SED). According to multiplex PCR, 99 isolates (53.7%) from dogs with pyoderma and 97 (90.8%) from healthy dogs possessed one or more se genes. There was no significant difference regarding ses between dogs with pyoderma and healthy dogs. Therefore, SEs may not be a direct virulence factor in pyoderma.

  7. Protective effects of bacterial osmoprotectant ectoine on bovine erythrocytes subjected to staphylococcal alpha-haemolysin.

    Science.gov (United States)

    Bownik, Adam; Stępniewska, Zofia

    2015-06-01

    Ectoine (ECT) is a bacterial compatible solute with documented protective action however no data are available on its effects on various cells against bacterial toxins. Therefore, we determined the in vitro influence of ECT on bovine erythrocytes subjected to staphylococcal α-haemolysin (HlyA). The cells exposed to HlyA alone showed a distinct haemolysis and reduced glutathione (GSH)/oxidised glutathione (GSSG) level, however the toxic effects were attenuated in the combinations of HlyA + ECT suggesting ECT-induced protection of erythrocytes from HlyA.

  8. Effect of the ionizing radiation on the Staphylococcal enterotoxins and Staphylococcus aureus in mechanically deboned chicken meat

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    Gomes, Heliana de Azevedo; Rocha, Claudia Regina Gomes; Roque, Claudio Vitor [Laboratorio de Pocos de Caldas (LAPOC/CNEN-MG), MG (Brazil)]. E-mail: hgomes@cnen.gov.br; crgrocha@gmail.com; cvroque@cnen.gov.br; Pereira, Jose Luiz [Universidade Estadual de Campinas, SP (Brazil). Dept. de Ciencias de Alimentos]. E-mail: pereira@fea.unicamp.br

    2007-07-01

    Ionizing radiation was employed in order to reduce contamination of mechanically deboned chicken meat (MDCM) by Staphylococcus aureus and also to evaluate the effect of the ionizing radiation on the staphylococcal enterotoxins. The microbiological analyses were carried out on MDCM refrigerated (+ 2 deg C) for 12 days. Irradiation treatments of MDCM with doses of 3.0 kGy and 4.0 kGy were capable of reducing the contamination by Staphylococcus aureus and reducing the amounts of staphylococcal enterotoxin, which might be present in this raw material. (author)

  9. Expression and production of staphylococcal enterotoxin C is substantially reduced in milk.

    Science.gov (United States)

    Valihrach, Lukas; Alibayov, Babek; Zdenkova, Kamila; Demnerova, Katerina

    2014-12-01

    Staphylococcal food poisoning is a global problem. The gene encoding enterotoxin C (sec) has been reported several times as the most frequent enterotoxin gene identified in food poisoning cases caused by contaminated milk. In this study, the expression of sec was examined during the growth of Staphylococcus aureus in milk compared to routinely used laboratory media. Additionally, expression of several regulatory genes (sarA, saeS, codY, srrA, rot, hld, agrA, sigB) and other five enterotoxin genes (sea, seg, seh, sek, sel) were observed. It has been well established for that S. aureus is able to grow in milk and we found significantly reduced expression of sec in milk compared to the laboratory medium (P enterotoxin genes and its regulation in milk. The milk environment dramatically changed the expression profiles of several enterotoxin genes although staphylococcal growth was not affected at all. The mechanism of the reduction may be explained by downregulation of the agr system, although other factors are expected to be involved. The constituent of milk causing the inhibitory effect remains unidentified.

  10. Evaluation of Potential Effects of NaCl and Sorbic Acid on Staphylococcal Enterotoxin A Formation

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    Nikoleta Zeaki

    2015-09-01

    Full Text Available The prophage-encoded staphylococcal enterotoxin A (SEA is recognized as the main cause of staphylococcal food poisoning (SFP, a common foodborne intoxication disease, caused by Staphylococcus aureus. Studies on the production of SEA suggest that activation of the SOS response and subsequent prophage induction affect the regulation of the sea gene and the SEA produced, increasing the risk for SFP. The present study aims to evaluate the effect of NaCl and sorbic acid, in concentrations relevant to food production, on SOS response activation, prophage induction and SEA production. The impact of stress was initially evaluated on steady state cells for a homogenous cell response. NaCl 2% was found to activate the SOS response, i.e., recA expression, and trigger prophage induction, in a similar way as the phage-inducer mitomycin C. In contrast, sorbic acid decreased the pH of the culture to a level where prophage induction was probably suppressed, even when combined with NaCl stress. The impact of previous physiological state of the bacteria was also addressed on cells pre-exposed to NaCl, and was found to potentially affect cell response upon exposure to further stress. The results obtained highlight the possible SFP-related risks arising from the use of preservatives during food processing.

  11. Staphylococcal enterotoxins in the Etiopathogenesis of Mucosal Autoimmunity within the Gastrointestinal Tract

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    MaryAnn Principato

    2014-04-01

    Full Text Available The staphylococcal enterotoxins (SEs are the products of Staphylococcus aureus and are recognized as the causative agents of classical food poisoning in humans following the consumption of contaminated food. While illness evoked by ingestion of the SE or its producer organism in tainted food are often self-limited, our current understanding regarding the evolution of S. aureus provokes the utmost concern. The organism and its associated toxins, has been implicated in a wide variety of disease states including infections of the skin, heart, sinuses, inflammatory gastrointestinal disease, toxic shock, and Sudden Infant Death Syndrome. The intricate relationship between the various subsets of immunocompetent T cells and accessory cells and the ingested material found within the gastrointestinal tract present daunting challenges to the maintenance of immunologic homeostasis. Dysregulation of the intricate balances within this environment has the potential for extreme consequences within the host, some of which are long-lived. The focus of this review is to evaluate the relevance of staphylococcal enterotoxin in the context of mucosal immunity, and the underlying mechanisms that contribute to the pathogenesis of gastrointestinal autoimmune disease.

  12. Plant-Derived Polyphenols Interact with Staphylococcal Enterotoxin A and Inhibit Toxin Activity.

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    Yuko Shimamura

    Full Text Available This study was performed to investigate the inhibitory effects of 16 different plant-derived polyphenols on the toxicity of staphylococcal enterotoxin A (SEA. Plant-derived polyphenols were incubated with the cultured Staphylococcus aureus C-29 to investigate the effects of these samples on SEA produced from C-29 using Western blot analysis. Twelve polyphenols (0.1-0.5 mg/mL inhibited the interaction between the anti-SEA antibody and SEA. We examined whether the polyphenols could directly interact with SEA after incubation of these test samples with SEA. As a result, 8 polyphenols (0.25 mg/mL significantly decreased SEA protein levels. In addition, the polyphenols that interacted with SEA inactivated the toxin activity of splenocyte proliferation induced by SEA. Polyphenols that exerted inhibitory effects on SEA toxic activity had a tendency to interact with SEA. In particular, polyphenol compounds with 1 or 2 hexahydroxydiphenoyl groups and/or a galloyl group, such as eugeniin, castalagin, punicalagin, pedunculagin, corilagin and geraniin, strongly interacted with SEA and inhibited toxin activity at a low concentration. These polyphenols may be used to prevent S. aureus infection and staphylococcal food poisoning.

  13. A novel approach to eliminate detection of contaminating Staphylococcal species introduced during clinical testing

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    Ao, Wanyuan; Clifford, Adrianne; Corpuz, Maylene; Jenison, Robert

    2017-01-01

    We describe here a strategy that can distinguish between Staphylococcus species truly present in a clinical sample from contaminating Staphylococcus species introduced during the testing process. Contaminating Staphylococcus species are present at low levels in PCR reagents and colonize lab personnel. To eliminate detection of contaminants, we describe an approach that utilizes addition of sufficient quantities of either non-target Staphylococcal cells (Staphylococcus succinus or Staphylococcus muscae) or synthetic oligonucleotide templates to helicase dependent isothermal amplification reactions to consume Staphylococcus-specific tuf and mecA gene primers such that contaminating Staphylococcus amplification is suppressed to below assay limits of detection. The suppressor template DNA is designed with perfect homology to the primers used in the assay but an internal sequence that is unrelated to the Staphylococcal species targeted for detection. Input amount of the suppressor is determined by a mathematical model described herein and is demonstrated to completely suppress contaminating levels of Staphylococcus while not negatively impacting the appropriate clinical assay limit of detection. We have applied this approach to improve the specificity of detection of Staphylococcus species present in positive blood cultures using a chip-based array that produces results visible to the unaided eye. PMID:28225823

  14. Contamination of beef products with staphylococcal classical enterotoxins in Egypt and Saudi Arabia

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    Shawish, Reyad R.

    2016-04-01

    Full Text Available Food-borne pathogens are of high concern for public health and food safety. food poisoning is one of the most economically devastating types of food poisoning globally. The purpose of this study was to detect staphylococcal classical enterotoxins (SEs in processed beef from Kingdom of Saudi Arabia (KSA and Egypt. In the present investigation a total of 250 random processed meat samples (50 each of minced meat, beef burger, beef sausage, beef kofta and beef luncheon were collected from different super markets in the study area. Using conventional cultural methods, samples were cultured for isolation and identification of . Multiplex PCR was used to detect SEs of the classical type SEA, SEB, SEC and SED from isolates.The percentage presence of in minced meat, beef burger, beef sausage, beef kofta and beef luncheon was 38%, 22%, 30%, 32% and 12%, respectively. Multiplex PCR indicated that all examined samples contain different types of classical staphylococcal enterotoxins and only minced meat samples contained all four types of toxins. Multiplex PCR is efficient in detection of SEs from food and may be used in tracing of toxins to promote food hygiene. Implications of contamination of processed meat to food hygiene in the study area are highlighted.

  15. Staphylococcus aureus and Staphylococcal Food-Borne Disease: An Ongoing Challenge in Public Health

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    Jhalka Kadariya

    2014-01-01

    Full Text Available Staphylococcal food-borne disease (SFD is one of the most common food-borne diseases worldwide resulting from the contamination of food by preformed S. aureus enterotoxins. It is one of the most common causes of reported food-borne diseases in the United States. Although several Staphylococcal enterotoxins (SEs have been identified, SEA, a highly heat-stable SE, is the most common cause of SFD worldwide. Outbreak investigations have found that improper food handling practices in the retail industry account for the majority of SFD outbreaks. However, several studies have documented prevalence of S. aureus in many food products including raw retail meat indicating that consumers are at potential risk of S. aureus colonization and subsequent infection. Presence of pathogens in food products imposes potential hazard for consumers and causes grave economic loss and loss in human productivity via food-borne disease. Symptoms of SFD include nausea, vomiting, and abdominal cramps with or without diarrhea. Preventive measures include safe food handling and processing practice, maintaining cold chain, adequate cleaning and disinfection of equipment, prevention of cross-contamination in home and kitchen, and prevention of contamination from farm to fork. This paper provides a brief overview of SFD, contributing factors, risk that it imposes to the consumers, current research gaps, and preventive measures.

  16. Study of action of biological aerococcus аutosymbiont on the model of staphylococcal infection

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    Stepanskyi D.O.

    2017-06-01

    Full Text Available The paper presents data on the study of the antagonistic effect of aerococcus autosymbionts in experimental models of infections caused by staphylococci. To study the antagonistic action of aerococcus autosymbiont on staphylococcus, a model of chronic staphylococcal infection in white mice and rabbits was used. In staphylococcal experimental infection, aerococcus antagonistic action against staphylococcus was tested by subcutaneous injection on white mice. Aerococci survival under the skin was studied. Aerococcus autosymbionts introduced under the skin to staphylococcus in 5 hours and 3 hours after administration of the latter cause antagonist effect and inhibit the development of infiltrates. Similar results were obtained when introducing aerococcus autosymbionts in the focus of infection in 5 minutes, 5 minutes and 3 hours, 1 and 5 hours after infection. Aerococci introduced after 24 and 48 hours shortened terms of disease manifestations as compared to the control group of mice. The therapeutic effect of aerococcus autosymbionts was tested on the experimental model of burns and wounds infected with staphylococcus. In animals with wounds treated with aerococcus autostrains the number of pathogenic staphylococci was 10 times less than in the control group. According to our observations aerococcus autosymbionts showed no irritant effect when applied on the wound surface, helped its healing, sharply reduced the percentage of staphylococcus inoculation from wound secretions.

  17. Interferon-γ Protects from Staphylococcal Alpha Toxin-Induced Keratinocyte Death through Apolipoprotein L1.

    Science.gov (United States)

    Brauweiler, Anne M; Goleva, Elena; Leung, Donald Y M

    2016-03-01

    Staphylococcus aureus is a bacterial pathogen that frequently infects the skin, causing lesions and cell destruction through its primary virulence factor, alpha toxin. Here we show that interferon gamma (IFN-?) protects human keratinocytes from cell death induced by staphylococcal alpha toxin. We find that IFN-? prevents alpha toxin binding and reduces expression of the alpha toxin receptor, a disintegrin and metalloproteinase 10 (ADAM10). We determine that the mechanism for IFN-?-mediated resistance to alpha toxin involves the induction of autophagy, a process of cellular adaptation to sublethal damage. We find that IFN-? potently stimulates activation of the primary autophagy effector, light chain 3 (LC3). This process is dependent on upregulation of apolipoprotein L1. Depletion of apolipoprotein L1 by small interfering RNA significantly increases alpha toxin-induced lethality and inhibits activation of light chain 3. We conclude that IFN-? plays a significant role in protecting human keratinocytes from the lethal effects of staphylococcal alpha toxin through apolipoprotein L1-induced autophagy.

  18. Effect of incubation atmosphere on the production and composition of staphylococcal biofilms.

    Science.gov (United States)

    Asai, Kentaro; Yamada, Keiko; Yagi, Tetsuya; Baba, Hisashi; Kawamura, Ichiro; Ohta, Michio

    2015-01-01

    Staphylococcus aureus and Staphylococcus epidermidis are pathogenic bacteria that often cause invasive infections in humans. In this study, we characterized the composition and growth characteristics of staphylococcal biofilms under various incubation atmospheres. We assessed the effect of incubation atmosphere (aerobic, 5% CO2, anaerobic, and microaerobic) on the biofilm production capabilities of S. aureus strains isolated from healthy volunteers and from patients with catheter-related bloodstream infection. In addition, the composition of S. aureus and S. epidermidis biofilms was determined by assessment of biofilm degradation after treatment with DNase I, proteinase K, and dispersin B. The strains obtained from healthy volunteers and patients showed similar biofilm formation capabilities. Biofilms of S. aureus were rich in proteins when developed under ambient atmospheric conditions, 5% CO2, and microaerobic condition, whereas S. epidermidis biofilms contained large amounts of poly-β (1, 6)-N-acetyl-D-glucosamine when developed under ambient atmospheric conditions and microaerobic condition. The biofilm-producing capability of S. epidermidis was considerably higher than that of S. aureus under aerobic condition. Staphylococcal isolates obtained from healthy individuals and patients with catheter-related infections have similar biofilm-forming capabilities. Under microaerobic conditions, S. aureus and S. epidermidis form protein-rich and poly-β (1, 6)-N-acetyl-D-glucosamine-rich biofilms, respectively. These components may play an important role in the development of biofilms inside the body and may be the target molecules to prevent catheter-related infections caused by these organisms.

  19. Staphylococcus aureus and staphylococcal food-borne disease: an ongoing challenge in public health.

    Science.gov (United States)

    Kadariya, Jhalka; Smith, Tara C; Thapaliya, Dipendra

    2014-01-01

    Staphylococcal food-borne disease (SFD) is one of the most common food-borne diseases worldwide resulting from the contamination of food by preformed S. aureus enterotoxins. It is one of the most common causes of reported food-borne diseases in the United States. Although several Staphylococcal enterotoxins (SEs) have been identified, SEA, a highly heat-stable SE, is the most common cause of SFD worldwide. Outbreak investigations have found that improper food handling practices in the retail industry account for the majority of SFD outbreaks. However, several studies have documented prevalence of S. aureus in many food products including raw retail meat indicating that consumers are at potential risk of S. aureus colonization and subsequent infection. Presence of pathogens in food products imposes potential hazard for consumers and causes grave economic loss and loss in human productivity via food-borne disease. Symptoms of SFD include nausea, vomiting, and abdominal cramps with or without diarrhea. Preventive measures include safe food handling and processing practice, maintaining cold chain, adequate cleaning and disinfection of equipment, prevention of cross-contamination in home and kitchen, and prevention of contamination from farm to fork. This paper provides a brief overview of SFD, contributing factors, risk that it imposes to the consumers, current research gaps, and preventive measures.

  20. Artificial biofilms establish the role of matrix interactions in staphylococcal biofilm assembly and disassembly.

    Science.gov (United States)

    Stewart, Elizabeth J; Ganesan, Mahesh; Younger, John G; Solomon, Michael J

    2015-08-14

    We demonstrate that the microstructural and mechanical properties of bacterial biofilms can be created through colloidal self-assembly of cells and polymers, and thereby link the complex material properties of biofilms to well understood colloidal and polymeric behaviors. This finding is applied to soften and disassemble staphylococcal biofilms through pH changes. Bacterial biofilms are viscoelastic, structured communities of cells encapsulated in an extracellular polymeric substance (EPS) comprised of polysaccharides, proteins, and DNA. Although the identity and abundance of EPS macromolecules are known, how these matrix materials interact with themselves and bacterial cells to generate biofilm morphology and mechanics is not understood. Here, we find that the colloidal self-assembly of Staphylococcus epidermidis RP62A cells and polysaccharides into viscoelastic biofilms is driven by thermodynamic phase instability of EPS. pH conditions that induce phase instability of chitosan produce artificial S. epidermidis biofilms whose mechanics match natural S. epidermidis biofilms. Furthermore, pH-induced solubilization of the matrix triggers disassembly in both artificial and natural S. epidermidis biofilms. This pH-induced disassembly occurs in biofilms formed by five additional staphylococcal strains, including three clinical isolates. Our findings suggest that colloidal self-assembly of cells and matrix polymers produces biofilm viscoelasticity and that biofilm control strategies can exploit this mechanism.

  1. Skin regeneration with conical and hair follicle structure of deep second-degree scalding injuries via combined expression of the EPO receptor and beta common receptor by local subcutaneous injection of nanosized rhEPO

    Directory of Open Access Journals (Sweden)

    Ebert S

    2012-03-01

    Full Text Available Augustinus Bader1, Sabine Ebert1, Shibashish Giri1, Mathias Kremer2, Shuhua Liu2, Andreas Nerlich5, Christina I Günter³, Dagmar U Smith4, Hans-Günther Machens2,31Department of Applied Stem Cell Biology and Cell Techniques, Centre for Biotechnology and Biomedicine, University of Leipzig, Leipzieg, 2Department of Plastic and Hand Surgery, University of Lübeck, Lübeck, 3Department of Plastic and Hand Surgery, Technische Universität München, Munich, 4Münchner Studienzentrum, Technische Universität München, Munich, 5Institute of Pathology, Klinikum München-Bogenhausen, Munich, GermanyBackground: Acceleration of skin regeneration is still an unsolved problem in the clinical treatment of patients suffering from deep burns and scalds. Although erythropoietin (EPO has a protective role in a wide range of organs and cells during ischemia and after trauma, it has been recently discovered that EPO is not tissue-protective in the common β subunit receptor (βCR knockout mouse. The protective capacity of EPO in tissue is mediated via a heteroreceptor complex comprising both the erythropoietin receptor (EPOR and βCR. However, proof of coexpression of these heterogenic receptors in regenerating skin after burns is still lacking.Methods: To understand the role of nanosized recombinant human erythropoietin (rhEPO in wound healing, we investigated the effects of subcutaneous injections of EPO on skin regeneration after deep second-degree scalding injuries. Our aim was to determine if joint expression of EPOR and βCR is a prerequisite for the tissue-protective effect of rhEPO. The efficiency in wound regeneration in a skin scalding injury mouse model was examined. A deep second-degree dermal scald injury was produced on the backs of 20 female Balb/c mice which were subsequently randomized to four experimental groups, two of which received daily subcutaneous injections of rhEPO. At days 7 and 14, the mice were sacrificed and the effects of rhEPO were

  2. 右美托咪定对严重烫伤大鼠心肌细胞PERK信号通路的影响%Effect of dexmedetomidine on PERK signaling pathway in cardiomyocytes of rats with severe scald

    Institute of Scientific and Technical Information of China (English)

    王惠枢; 张良成; 徐世元

    2016-01-01

    Objective To investigate the effect of dexmedetomidine on protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling pathway in cardiomyocytes of the rats with severe scald.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 220-280 g,were randomly divided into 3 groups (n =8 each) using a random number table:control group (group C),severe scald group (group S),and scald + dexmedetomidine group (group D).Thirty percent of the total body surface area was shaved on the back and then exposed to 94 ℃ water for 12 s to establish the model of 3rd degree scald.Dexmedetomidine 30 μg/kg (2 μg/ml) was intraperitoneally injected immediately after scald in group D.Myocardial specimens were obtained at 12 h after scald for examination of the pathological changes and for determination of cell apoptosis and expression of C/EBP-homologous protein (CHOP),PERK,and phosphorylated PERK (p-PERK) by Western blot.The apoptosis index and p-PERK/PERK ratio were calculated.Results Compared with group C,the apoptosis index was significantly increased,the expression of CHOP,PERK and p-PERK was significantly up-regulated,and the p-PERK/PERK ratio was significantly increased in S and D groups (P<0.05).Compared with group S,the apoptosis index was significantly decreased,the expression of CHOP,PERK and p-PERK was significantly down-regulated,and the p-PERK/PERK ratio was significantly decreased (P<0.05),and the pathological changes of myocardium were significantly attenuated in group D.Conclusion The mechanism by which dexmedetomidine inhibits apoptosis in cardiomyocytes is related to inhibition of PERK signaling pathway in the rats with severe scald.%目的 探讨右美托咪定对严重烫伤大鼠心肌细胞蛋白激酶R样内质网激酶(PERK)信号通路的影响.方法 健康成年雄性SD大鼠24只,体重220~ 280 g,采用随机数字表法分为3组(n=8):对照组(C组)、严重烫伤组(S组)、烫伤+右美托咪定组(D组).将大鼠背部浸入94

  3. Xanthomonas albilineans OmpA1 appears to be functionally modular and both the OMC and C-like domains are necessary for leaf scald disease of sugarcane.

    Science.gov (United States)

    Fleites, Laura A; Mensi, Imène; Gargani, Daniel; Zhang, Shujian; Rott, Philippe; Gabriel, Dean W

    2013-10-01

    Several EZ-Tn5 insertions in gene locus XALc_0557 (OmpA1) of the sugarcane leaf scald pathogen Xanthomonas albilineans XaFL07-1 were previously found to strongly affect pathogenicity and endophytic stalk colonization. XALc_0557 has a predicted OmpA N-terminal outer membrane channel (OMC) domain and an OmpA C-like domain. Further analysis of mutant M468, with an EZ-Tn5 insertion in the upstream OMC domain coding region, revealed impaired epiphytic and endophytic leaf survival, impaired resistance to sodium dodecyl sulfate (SDS), structural defects in the outer membrane (OM), and hyperproduction of OM vesicles. Cloned full-length XALc_0557 complemented M468 for all phenotypes tested, including pathogenicity, resistance to SDS, and ability to survive both endophytically and epiphytically. Another construct, pCT47.3, which expressed only the C-like domain of XALc_0557, restored resistance to SDS in M468 but failed to complement any other mutant phenotype, indicating that the C-like domain functioned independently of the OMC domain to help maintain OM integrity. pCT47.3 also complemented pathogenicity, resistance to SDS, and stalk colonization in mutant M1152, which carries an EZ-Tn5 insert in the C-like coding region, indicating that both predicted domains are modular and necessary but neither is sufficient for X. albilineans pathogenicity, endophytic survival in, and epiphytic survival on sugarcane.

  4. COMPARISON OF THE EFFECTS OF AGING AND IL-6 ON THE HEPATIC INFLAMMATORY RESPONSE IN TWO MODELS OF SYSTEMIC INJURY: SCALD INJURY VERSUS I.P. LPS ADMINISTRATION

    Science.gov (United States)

    Gomez, Christian R.; Nomellini, Vanessa; Baila, Horea; Oshima, Kiyoko; Kovacs, Elizabeth J.

    2011-01-01

    Regardless of age, a marked elevation in circulating IL-6 levels correlates with increased mortality after injury or an inflammatory challenge. We previously reported that aged IL-6 knockout mice given LPS have improved survival and reduced inflammatory response than LPS-treated aged wild type (WT) mice. Herein, we analyzed the effects of aging and IL-6 on the hepatic inflammatory response in two models of systemic injury: dorsal scald (burn) injury versus intraperitoneal LPS administration. At 24 h after burn injury, circulating alanine aminotransferase and hepatic neutrophil accumulation were comparable regardless of age or IL-6 deficiency. However, at this same time point, these indicators of liver damage, in addition to hepatic levels of KC, a neutrophil chemoattractant, were increased in aged WT mice given LPS relative to young WT mice given LPS. The hepatic injury was drastically reduced in aged IL-6 knockout mice given LPS as compared with LPS-exposed aged WT mice. Our results suggest that the nature of the insult will determine the degree of remote injury in aged animals. In addition, the role of IL-6 as a contributing factor of tissue injury may be insult specific. PMID:18636046

  5. Comparison of the effects of aging and IL-6 on the hepatic inflammatory response in two models of systemic injury: scald injury versus i.p. LPS administration.

    Science.gov (United States)

    Gomez, Christian R; Nomellini, Vanessa; Baila, Horea; Oshima, Kiyoko; Kovacs, Elizabeth J

    2009-02-01

    Regardless of age, a marked elevation in circulating IL-6 levels correlates with increased mortality after injury or an inflammatory challenge. We previously reported that aged IL-6 knockout mice given LPS have improved survival and reduced inflammatory response than LPS-treated aged wild type (WT) mice. Herein, we analyzed the effects of aging and IL-6 on the hepatic inflammatory response in two models of systemic injury: dorsal scald (burn) injury versus intraperitoneal LPS administration. At 24 h after burn injury, circulating alanine aminotransferase and hepatic neutrophil accumulation were comparable regardless of age or IL-6 deficiency. However, at this same time point, these indicators of liver damage, in addition to hepatic levels of KC, a neutrophil chemoattractant, were increased in aged WT mice given LPS relative to young WT mice given LPS. The hepatic injury was drastically reduced in aged IL-6 knockout mice given LPS as compared with LPS-exposed aged WT mice. Our results suggest that the nature of the insult will determine the degree of remote injury in aged animals. In addition, the role of IL-6 as a contributing factor of tissue injury may be insult specific.

  6. Identification of staphylococcal species based on variations in protein sequences (mass spectrometry) and DNA sequence (sodA microarray).

    Science.gov (United States)

    Kooken, Jennifer; Fox, Karen; Fox, Alvin; Altomare, Diego; Creek, Kim; Wunschel, David; Pajares-Merino, Sara; Martínez-Ballesteros, Ilargi; Garaizar, Javier; Oyarzabal, Omar; Samadpour, Mansour

    2014-02-01

    This report is among the first using sequence variation in newly discovered protein markers for staphylococcal (or indeed any other bacterial) speciation. Variation, at the DNA sequence level, in the sodA gene (commonly used for staphylococcal speciation) provided excellent correlation. Relatedness among strains was also assessed using protein profiling using microcapillary electrophoresis and pulsed field electrophoresis. A total of 64 strains were analyzed including reference strains representing the 11 staphylococcal species most commonly isolated from man (Staphylococcus aureus and 10 coagulase negative species [CoNS]). Matrix assisted time of flight ionization/ionization mass spectrometry (MALDI TOF MS) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC ESI MS/MS) were used for peptide analysis of proteins isolated from gel bands. Comparison of experimental spectra of unknowns versus spectra of peptides derived from reference strains allowed bacterial identification after MALDI TOF MS analysis. After LC-MS/MS analysis of gel bands bacterial speciation was performed by comparing experimental spectra versus virtual spectra using the software X!Tandem. Finally LC-MS/MS was performed on whole proteomes and data analysis also employing X!tandem. Aconitate hydratase and oxoglutarate dehydrogenase served as marker proteins on focused analysis after gel separation. Alternatively on full proteomics analysis elongation factor Tu generally provided the highest confidence in staphylococcal speciation.

  7. Shortening the Antibiotic Course for the Treatment of Neonatal Coagulase-Negative Staphylococcal Sepsis : Fine with Three Days?

    NARCIS (Netherlands)

    Hemels, Marieke A. C.; van den Hoogen, Agnes; Verboon-Maciolek, Malgorzata A.; Fleer, Andre; Krediet, Tannette G.

    2012-01-01

    Background: The incidence of coagulase-negative staphylococcal (CoNS) sepsis is high in neonatal intensive care units (NICUs) and treatment significantly adds to the antibiotic pressure, increasing the threat of resistance. Because infants recover within 24-48 h, blood cultures are negative within 4

  8. Structural variations of staphylococcal cassette chromosome mec Type IVa in Staphylococcus aureus clonal complex 8 and unrelated lineages

    DEFF Research Database (Denmark)

    Damborg, Peter Panduro; Bartels, Mette Damkjær; Boye, Kit

    2011-01-01

    PCR mapping of staphylococcal cassette chromosome mec type IVa and adjacent mobile elements in 94 methicillin-resistant Staphylococcus aureus (MRSA) strains identified two primary structures (A and B) that could be further classified into two (A1 and A2) and five (B1 to B5) variants, primarily...

  9. Probabilistic risk model for staphylococcal intoxication from pork-based food dishes prepared in food service establishments in Korea.

    Science.gov (United States)

    Kim, Hyun Jung; Griffiths, Mansel W; Fazil, Aamir M; Lammerding, Anna M

    2009-09-01

    Foodborne illness contracted at food service operations is an important public health issue in Korea. In this study, the probabilities for growth of, and enterotoxin production by, Staphylococcus aureus in pork meat-based foods prepared in food service operations were estimated by the Monte Carlo simulation. Data on the prevalence and concentration of S. aureus as well as compliance to guidelines for time and temperature controls during food service operations were collected. The growth of S. aureus was initially estimated by using the U.S. Department of Agriculture's Pathogen Modeling Program. A second model based on raw pork meat was derived to compare cell number predictions. The correlation between toxin level and cell number as well as minimum toxin dose obtained from published data was adopted to quantify the probability of staphylococcal intoxication. When data gaps were found, assumptions were made based on guidelines for food service practices. Baseline risk model and scenario analyses were performed to indicate possible outcomes of staphylococcal intoxication under the scenarios generated based on these data gaps. Staphylococcal growth was predicted during holding before and after cooking, and the highest estimated concentration (4.59 log CFU/g for the 99.9th percentile value) of S. aureus was observed in raw pork initially contaminated with S. aureus and held before cooking. The estimated probability for staphylococcal intoxication was very low, using currently available data. However, scenario analyses revealed an increased possibility of staphylococcal intoxication when increased levels of initial contamination in the raw meat, andlonger holding time both before and after cooking the meat occurred.

  10. Staphylococcal enterotoxin A gene-carrying Staphylococcus aureus isolated from foods and its control by crude alkaloid from papaya leaves.

    Science.gov (United States)

    Handayani, Lita; Faridah, Didah Nur; Kusumaningrum, Harsi D

    2014-11-01

    Staphylococcus aureus is a known pathogen causing intoxication by producing enterotoxins in food. Staphylococcal enterotoxin A is one of the enterotoxins commonly implicated in staphylococcal food poisoning. The ability of crude alkaloid extract from papaya leaves to inhibit the growth of S. aureus and staphylococcal enterotoxin A synthesis was investigated. Staphylococcal enterotoxin A gene-carrying S. aureus was isolated from raw milk and ready-to-eat foods. Crude alkaloid was extracted from ground, dried papaya leaves using ultrasonic-assisted extraction, and a MIC of the alkaloid was determined by the broth macrodilution method. Furthermore, S. aureus isolate was exposed to the crude alkaloid extract at one- and twofold MIC, and the expression of sea was subsequently analyzed using a quantitative reverse transcription real-time PCR. Ten isolates of S. aureus were obtained, and nine of those isolates were sea carriers. The yield of crude alkaloid extract was 0.48 to 1.82% per dry weight of papaya leaves. A MIC of crude alkaloid to S. aureus was 0.25 mg/ml. After exposure to the alkaloid at 0.25 and 0.5 mg/ml for 2 h, a significant increase in cycle threshold values of sea was observed. The sea was expressed 29 and 41 times less when S. aureus was exposed to crude alkaloid at one- and twofold MIC, respectively. This study revealed that crude alkaloid of papaya leaves could control staphylococcal enterotoxin A gene-carrying S. aureus by suppressing the expression of sea, in addition to the ability to inhibit the growth of S. aureus. The expression of sea was successfully quantified.

  11. Cross-talk between Staphylococcus aureus and other staphylococcal species via the agr quorum sensing system

    Directory of Open Access Journals (Sweden)

    Jaime Canovas de la Nuez

    2016-11-01

    Full Text Available Staphylococci are associated with both humans and animals. While most are non-pathogenic colonizers, Staphylococcus aureus is an opportunistic pathogen capable of causing severe infections. S. aureus virulence is controlled by the agr quorum sensing system responding to secreted auto-inducing peptides (AIPs sensed by AgrC, a two component histidine kinase. agr loci are found also in other staphylococcal species and for S. epidermidis, the encoded AIP represses expression of agr regulated virulence genes in S. aureus. In this study we aimed to determine how other staphylococci affect S. aureus agr, and if such interaction may point to new anti-virulence candidates to target S. aureus infections. Here we show that culture supernatants of 37 out of 52 staphylococcal isolates representing 17 different species inhibit S. aureus agr. The dog pathogen, S. schleiferi, expressed the most potent inhibitory activity and was active against all four agr classes found in S. aureus. By employing a S. aureus strain encoding a constitutively active AIP receptor we show that the activity is mediated via agr. Subsequent cloning and heterologous expression of the S. schleiferi AIP in S. aureus demonstrated that this molecule was likely responsible for the inhibitory activity, and further proof was provided when pure synthetic S. schleiferi AIP was able to completely abolish agr induction of an S. aureus reporter strain. To assess impact on S. aureus virulence, we co-inoculated S. aureus and S. schleiferi in vivo in the Galleria Mellonella wax moth larva, and found that expression of key S. aureus virulence factors was abrogated. Our data show that the S. aureus agr locus is highly responsive to other staphylococcal species suggesting that agr is an inter-species communication system. Based on these results we speculate that interactions between S. aureus and other colonizing staphylococci will significantly influence the ability of S. aureus to cause infection, and

  12. ASSESSMENT OF THE RISK OF RAW MILK CONSUMPTION RELATED TO STAPHYLOCOCCAL FOOD POISONING

    Directory of Open Access Journals (Sweden)

    Elsa Helena Walter Santana

    2010-10-01

    Full Text Available This study aimed to identify the risks of staphylococcal food poisoning due to the consumption of raw milk. Fifty-one farms in Londrina (PR and 50 in Pelotas (RS were analyzed, to determinethe population of coagulase-positive staphylococci (UFC/ mL, as well as to verify the ability of producing Staphylococcal Enterotoxin A (SEA by immunodifusion (OSP, the presence of the gene for the production of SEA (PCR in the cultures, and the research of enterotoxin (SEA to SEE in milk samples using ELISA commercial kit. Considering the 101 farms analyzed, 19 (18.8% presented coagulase-positive staphylococci count above 105 UFC/mL. For the evaluation of the enterotoxigenic ability (SEA by the OSP technique, six cultures coagulase-positive (5.5% were positive to the test and identified as S. aureus. From the coagualse-negative sample, one (5.5% was OSP positive. For the evaluation of the presence of the gene for EEA synthesis, 51 cultures of staphylococci were tested. From this total, 14 (27.45% presented the gene, and from that, only 5 (9.81% cultures were capable of expressing it in the technique of the OSP. The morphologiccharacteristic of the evaluated cultures that had enterotoxigenic capacity, from the 14 (33,3% cultures that presented the gene for EEA production, 05 (11.9% were characterized as typical cultures of S.aureus in Baird Parker agar. All the 12 milk samples studied for the presence of EEA to EEE in milk were negative. Thus, it can be concluded that there is extensive contamination of raw milk for staphylococci coagulase, however, most of the isolated strains were not enterotoxigenic or did not express such a characteristic. Only 9.81% of the tested colonies expressed the gene and effectivelyproduced SEA. None of the samples had sufficient counts to produce detectable amounts of SEA. The milk samples did not present risk to cause staphylococcal food poisoning if consumed in natura until the collection moment.

  13. Staphylococcal Infections

    Science.gov (United States)

    ... you should be familiar with include the following: Cellulitis is a bacterial skin infection that first affects the outer layers of the ... skin. Although other types of bacteria can cause cellulitis, Saureus ... may diagnose the infection by examining the area. The doctor may take ...

  14. Phytochemical and anti-staphylococcal biofilm assessment of Dracaena draco L. Spp. draco resin

    Directory of Open Access Journals (Sweden)

    V. Di Stefano

    2014-01-01

    Full Text Available Background: Dracaena draco L. ssp. draco is known as the "dragon′s blood tree" and it is endemic from the Canary Islands and Morocco. Objective: Carry out phytochemical investigation of acetonic extracts of red resin obtained from the trunk of D. draco, to obtain to the isolation of the most abundant resin constituents, belonging to the class of flavonoids: flavans, along with homoisoflavans and homoisoflavanones. Materials and Methods: The structures of the isolated compounds were established by Nuclear Magnetic Resonance (NMR and mass spectrometry data and comparison with literature data. The acetonic extract was evaluated for its anti-staphylococcal properties against two reference strains. Results: The acetonic extracts resulted inactive at the maximum tested concentration of 1000 μg/ml against free living forms of tested staphylococci, but they showed a very interesting activity in the prevention of a biofilm formation at a concentration equal to 200 μg/ml against S. aureus ATCC 25923.

  15. 超抗原SEA研究进展%Progress of Superantigenic Staphylococcal Enterotoxin A Researching

    Institute of Scientific and Technical Information of China (English)

    薛华; 何生

    2001-01-01

    超抗原是一类只需极低浓度(1~10 ng/ml) 即可激活大量的淋巴细胞克隆,产生极强的应答效应的抗原性物质。葡萄球菌肠毒素A(staphylococcal enterotoxin A,SEA)是金黄色葡萄球菌产生的葡萄球菌肠毒素家族中一员,是最有效的T细胞活化因子之一。研究发现,SEA可通过刺激T细胞增殖,促进IL-2,TNF-α,IFN-γ等淋巴因子的产生,从而发挥抗肿瘤免疫治疗作用。

  16. Structural basis for Zn2+-dependent intercellular adhesion in staphylococcal biofilms.

    Science.gov (United States)

    Conrady, Deborah G; Wilson, Jeffrey J; Herr, Andrew B

    2013-01-15

    Staphylococcal bacteria, including Staphylococcus epidermidis and Staphylococcus aureus, cause chronic biofilm-related infections. The homologous proteins Aap and SasG mediate biofilm formation in S. epidermidis and S. aureus, respectively. The self-association of these proteins in the presence of Zn(2+) leads to the formation of extensive adhesive contacts between cells. This study reports the crystal structure of a Zn(2+) -bound construct from the self-associating region of Aap. Several unusual structural features include elongated β-sheets that are solvent-exposed on both faces and the lack of a canonical hydrophobic core. Zn(2+)-dependent dimers are observed in three distinct crystal forms, formed via pleomorphic coordination of Zn(2+) in trans across the dimer interface. These structures illustrate how a long, flexible surface protein is able to form tight intercellular adhesion sites under adverse environmental conditions.

  17. Multiplexed Electrochemical Detection of Yersinia Pestis and Staphylococcal Enterotoxin B using an Antibody Microarray

    Directory of Open Access Journals (Sweden)

    Jason Wojciechowski

    2010-04-01

    Full Text Available The CombiMatrix antibody microarray is a versatile, sensitive detection platform based on the generation and transduction of electrochemical signals following antigen binding to surface antibodies. The sensor chip described herein is comprised of microelectrodes coupled to an adjacent bio-friendly matrix coated with antibodies to the biological pathogens Yersinia pestis and Bacillus anthracis, and the bacterial toxin staphylococcal enterotoxin B (SEB. Using this system, we were able to detect SEB and inactivated Y. pestis individually as well as in two-plex assays at concentrations as low as 5 pg/mL and 106 CFU/mL, respectively. We also introduce super avidin-biotin system (SABS as a viable and effective means to enhance assay signal responses and lower detection limits. Together these technologies represent substantial advances in point-of-care and point-of-use detection applications.

  18. Safety and efficacy of moxifloxacin monotherapy for treatment of orthopedic implant-related staphylococcal infections.

    Science.gov (United States)

    San Juan, Rafael; Garcia-Reyne, Ana; Caba, Pedro; Chaves, Fernando; Resines, Carlos; Llanos, Fernando; López-Medrano, Francisco; Lizasoain, Manuel; Aguado, Jose Maria

    2010-12-01

    The rifampin-ciprofloxacin combination is recommended for treatment of orthopedic implant-related staphylococcal infections to avoid the emergence of ciprofloxacin resistance; however, the efficacy of this combination is limited by the tolerability problems associated with the use of rifampin. Moxifloxacin is a quinolone up to 10 times more active against staphylococci than ciprofloxacin and the risk of resistance development during monotherapy against staphylococci is theoretically lower for moxifloxacin, but information regarding its use in bone infections is lacking. The aim of the present study was to evaluate the safety and clinical efficacy of moxifloxacin monotherapy in patients with orthopedic implant-related staphylococcal infections. From June 2006 to April 2009, all patients with culture-proven infection by quinolone-sensitive staphylococcal strains associated with orthopedic implants at our institution were included in a management protocol that mostly included specific surgery, 1 to 2 weeks of an intravenous course of cloxacillin-cefazolin or vancomycin, and long-term therapy with moxifloxacin (400 mg/day for 3 months). Cure was defined as (i) a lack of clinical signs and symptoms of infection, (ii) a C-reactive protein level less than 5 mg/liter, and (iii) absence of radiological signs of loosening or infection at the latest follow-up visit. Failure was defined as (i) persisting clinical and/or laboratory signs of infection or (ii) persisting or new isolation of the initial microorganism. A total of 48 patients with a median follow-up of 716 days (range, 102 to 1,613 days) were included in the study. Complete drug compliance was achieved in all but two patients (4.2%), who required drug discontinuation because of side effects (diarrhea and dizziness). No moxifloxacin-induced arrhythmia was reported. Twenty patients had joint prosthesis infections (5 acute-onset infections and 15 chronic infections), and 28 patients had osteosynthesis material

  19. Safety and Efficacy of Moxifloxacin Monotherapy for Treatment of Orthopedic Implant-Related Staphylococcal Infections ▿

    Science.gov (United States)

    San Juan, Rafael; Garcia-Reyne, Ana; Caba, Pedro; Chaves, Fernando; Resines, Carlos; Llanos, Fernando; López-Medrano, Francisco; Lizasoain, Manuel; Aguado, Jose Maria

    2010-01-01

    The rifampin-ciprofloxacin combination is recommended for treatment of orthopedic implant-related staphylococcal infections to avoid the emergence of ciprofloxacin resistance; however, the efficacy of this combination is limited by the tolerability problems associated with the use of rifampin. Moxifloxacin is a quinolone up to 10 times more active against staphylococci than ciprofloxacin and the risk of resistance development during monotherapy against staphylococci is theoretically lower for moxifloxacin, but information regarding its use in bone infections is lacking. The aim of the present study was to evaluate the safety and clinical efficacy of moxifloxacin monotherapy in patients with orthopedic implant-related staphylococcal infections. From June 2006 to April 2009, all patients with culture-proven infection by quinolone-sensitive staphylococcal strains associated with orthopedic implants at our institution were included in a management protocol that mostly included specific surgery, 1 to 2 weeks of an intravenous course of cloxacillin-cefazolin or vancomycin, and long-term therapy with moxifloxacin (400 mg/day for 3 months). Cure was defined as (i) a lack of clinical signs and symptoms of infection, (ii) a C-reactive protein level less than 5 mg/liter, and (iii) absence of radiological signs of loosening or infection at the latest follow-up visit. Failure was defined as (i) persisting clinical and/or laboratory signs of infection or (ii) persisting or new isolation of the initial microorganism. A total of 48 patients with a median follow-up of 716 days (range, 102 to 1,613 days) were included in the study. Complete drug compliance was achieved in all but two patients (4.2%), who required drug discontinuation because of side effects (diarrhea and dizziness). No moxifloxacin-induced arrhythmia was reported. Twenty patients had joint prosthesis infections (5 acute-onset infections and 15 chronic infections), and 28 patients had osteosynthesis material

  20. Efeitos metabólicos da glutamina em ratos submetidos à queimadura por água fervente (escaldadura Metabolic effects of glutamine in rats subjected to scald burn

    Directory of Open Access Journals (Sweden)

    Rômulo Cesar Costa Barbosa

    2003-12-01

    Full Text Available OBJETIVO: Investigar os efeitos metabólicos da L-glutamina (Gln em ratos anestesiados submetidos à queimadura por água fervente. MÉTODOS: Foram estudados vinte e quatro ratos Wistar machos, anestesiados, submetidos a queimaduras da pele dorsal após exposição à água quente (100ºC durante 10 segundos (30% de superfície corporal. Os ratos foram randomizados para receber, por gavagem, 2ml de água (G-1 ou igual volume de solução de Gln (0,5g/peso/dia (G-2. Amostras de tecido (pele sadia e queimada, músculo e fígado e sangue foram coletadas 24h (D1 e 48h (D2 pós-trauma para análise enzimática. RESULTADOS: A oferta de Gln induziu aumento significante nas concentrações de glicose na pele saudável em animais do G-2 no D2, e na pele queimada em G-2/D1. As concentrações de lactato também aumentaram significantemente em G-2/D1 no músculo (11,29 ± 1,25 mmol/g contra 7,43 ± 0,93 mmol/g - pPURPOSE: Investigate the metabolic effects of L- glutamine (Gln in rats subjected to scald burn. METHODS: Twenty-four anesthetized male Wistar rats were submitted to scald burn of dorsal skin secondary to exposure to hot water (100ºC for 10 sec (30% of body surface. Eighteen and 42h later rats were randomized to receive (by gavage 2ml of water (G-1 or equal volume (0,5g/Kg weight/day of Gln solution (G-2. Tissue and blood samples were collected at the end of 24h (D1 and 48h (D2 post burn for enzymatic analysis. RESULTS: Glucose concentrations were significantly increased in healthy skin in G-2/D2 and in burned skin in G-2/D1. Lactate concentrations were significantly increased in G-2/D1 in muscle (11,29 ± 1,25 mmol/g versus 7,43 ± 0,93 mmol/g - p<0.05 and in G-2/D2 subgroups in liver (7,68 ± 1,49 mmol/g versus 3,27 ±0,67 mmol/g - p<0.01 and healthy skin (5,30 ± 0,42 mmol/g versus 3,57 ± 0,38 mmol/g - p<0.05. Pyruvate concentrations were significantly decreased in G-2/D1 subgroups in muscle and liver and increased in healthy skin in G-2/D2

  1. Nasopharyngeal colonization elicits antibody responses to staphylococcal and pneumococcal proteins that are not associated with a reduced risk of subsequent carriage

    NARCIS (Netherlands)

    S.M.P.J. Prevaes (Sabine); W.J.B. van Wamel (Willem); C.P. de Vogel (Corné); R.H. Veenhoven (Reinier); A. van Gils (Anita); A.F. van Belkum (Alex); E.A. Sanders (Elisabeth); D. Bogaert (Debby)

    2012-01-01

    textabstractKnowledge of the immunological correlates of Staphylococcus aureus and Streptococcus pneumoniae colonization is required for the search for future protein vaccines. We evaluated natural antibody levels against pneumococcal and staphylococcal proteins in relation to previous bacterial

  2. Molecular Detection of Inducible Clindamycin Resistance among Staphylococcal Strains Isolated from Hospital Patients

    Directory of Open Access Journals (Sweden)

    Shadiyeh Abdollahi

    2013-04-01

    Full Text Available Background & Objectives: Macrolide, lincosamide and streptogramin B (MLSB antimicrobial agents are used in the treatment of staphylococcal infections. They prevent the microbial protein synthesis system through binding to 23 S rRNA. The aim of this study was to apply molecular methods to detect inducible clindamycin resistance genes among staphylococcal strains isolated from clinical specimens.   Methods : Two hundred staphylococcus strains were isolated from nose and throat swabs of patients in Toohid and Besat hospitals in Sanandaj . Antimicrobial susceptibilities of isolates were determined using disc diffusion method, agar screen test and D-Test. A multiplex PCR was performed using primers specific for erm (A, B, C, TR genes.   Results: Out of 200 isolates, 18.5 % were MRSA and 32% were MRCNS (methicillin resistant coagulase negative staphylococci. Of 80 erythromycin resistant isolates, 48 were coagulase negative and 32 were S. aureus. Among the 48 coagulase negative staphylococci (CONS isolates, 11.63% expressed the MLSB-inducible phenotypes. Using PCR, the frequency of different genes in the collection of isolates were as follows: ermA 5.41 % , erm B 5.41 % , and erm C 3.13%. The ermTR gene was negative in all isolates. Among the 32 S. aureus isolates, 9.38% expressed the MLSB-nducible phenotype. Using PCR, these isolates harbored erm A (2.22%, ermB (2.22%, ermC (2.22% and ermTR (2.22% .   Conclusion: This is the first study to show the rate of inducible clindamycin clinical isolates of staphylococci harboring erm genes in Sananadaj. It also demonstrated the frequency of erm genes was higher among CONS isolates than S. aureus. This data suggested the transfer of resistance gene from nonpathogenic to pathogenic strains is likely to happen. Therefore, screening and control of these resistance genes is recommended at clinical laboratories.

  3. Structural basis for pore-forming mechanism of staphylococcal α-hemolysin.

    Science.gov (United States)

    Sugawara, Takaki; Yamashita, Daichi; Kato, Koji; Peng, Zhao; Ueda, Junki; Kaneko, Jun; Kamio, Yoshiyuki; Tanaka, Yoshikazu; Yao, Min

    2015-12-15

    Staphylococcal α-hemolysin (α-HL) is a β-barrel pore-forming toxin (PFT) expressed by Staphylococcus aureus. α-HL is secreted as a water-soluble monomeric protein, which binds to target membranes and forms membrane-inserted heptameric pores. To explore the pore-forming mechanism of α-HL in detail, we determined the crystal structure of the α-HL monomer and prepore using H35A mutant and W179A/R200A mutant, respectively. Although the overall structure of the monomer was similar to that of other staphylococcal PFTs, a marked difference was observed in the N-terminal amino latch, which bent toward the prestem. Moreover, the prestem was fastened by the cap domain with a key hydrogen bond between Asp45 and Tyr118. Prepore structure showed that the transmembrane region is roughly formed with flexibility, although the upper half of the β-barrel is formed appropriately. Structure comparison among monomer, prepore and pore revealed a series of motions, in which the N-terminal amino latch released upon oligomerization destroys its own key hydrogen bond between Asp45-Tyr118. This action initiated the protrusion of the prestem. Y118F mutant and the N-terminal truncated mutant markedly decreased in the hemolytic activity, indicating the importance of the key hydrogen bond and the N-terminal amino latch on the pore formation. Based on these observations, we proposed a dynamic molecular mechanism of pore formation for α-HL.

  4. Comparative Effects of Food Preservatives on the Production of Staphylococcal Enterotoxin I from Staphylococcus aureus Isolate

    Directory of Open Access Journals (Sweden)

    Yanying Zhao

    2017-01-01

    Full Text Available Staphylococcal enterotoxin I (SEI is associated with staphylococcal food poisoning, but little is known about different food preservatives on the production of SEI. In this study, the effect of different food preservatives (sodium nitrite, polylysine, chitosan, and tea catechin on the bacteria growth, sei gene expression, and extracellular SEI production of Staphylococcus aureus isolate H4 was detected in tryptone soya broth (TSB culture. Our results showed that all of these preservatives depressed S. aureus H4 growth and the order of inhibitory effect was 0.8 g/L tea catechin > 6 g/L chitosan > 0.25 g/L polylysine > 0.4 g/L tea catechin > 0.15 g/L sodium nitrite. Furthermore, 0.25 g/L polylysine or 0.15 g/L sodium nitrite did not significantly alter sei gene transcription, while 6 g/L chitosan obviously increased the relative mRNA level of sei gene expression. 0.4 g/L tea catechin remarkably inhibited sei gene transcription. In addition, 0.15 g/L sodium nitrite and 6 g/L chitosan significantly enhanced SEI secretion. 0.25 g/L polylysine, especially 0.4 g/L tea catechin, sharply inhibited the level of SEI secretion. The results indicated that tea catechin not only suppressed Staphylococcus aureus growth, but also inhibited SEI production and secretion, suggesting that tea catechin may be better than sodium nitrite, polylysine, or chitosan for keeping the food from the contamination of SEI. These investigations would be useful for food industry to provide safer food products due to S. aureus enterotoxins-related control strategy.

  5. Effects and Mechanism of Chitosan Coatings in the Control of Superficial Scald in Apples%壳聚糖涂膜对苹果虎皮病防治效果与机理研究

    Institute of Scientific and Technical Information of China (English)

    吕新刚; 刘兴华; 蔡露阳

    2011-01-01

    The effects of chitosan coatings on the control of superficial scald in " Fuji" apples were investigated, and the dynamic changes in a-farnesene production and oxidation, the texture and function of cell membrane and the activity of enzyme related with browning were also measured to reveal the mechanism of chitosan coatings in the superficial scald control. The results indicated that by chitosan coatings, the respiration rate and the ethylene production were inhibited, the production of a-farnesene and the accumulation of conjugated trienes were reduced, membrane lipid peroxidation was weakened and the integrity of cell membrane was kept, and the increase of polyphenoloxidase ( PPO) activities was inhibited, the chitosan coating could effectively control the superficial scald in apples.%以红富士苹果为试材,研究了壳聚糖涂膜处理对苹果虎皮病的防治效果,并从α-法尼烯的生成及氧化、膜结构和功能及褐变相关酶活性变化3个方面探讨壳聚糖涂膜对贮藏过程中苹果生理变化的影响,以揭示其对虎皮病的作用机理.结果表明:壳聚糖涂膜可以抑制果实呼吸作用和乙烯的生成,降低α-法尼烯生成及氧化产物共轭三烯的积累,降低膜脂过氧化作用,较好地保持细胞膜的完整性,抑制PPO活性的升高;壳聚糖涂膜处理对虎皮病有明显的防治作用.

  6. Experimental Study on the Therapeutic Action of Salicylic Acid - g - chitosan Derivatives on Skin Scald%水杨酸-g-壳聚糖衍生物对皮肤溃疡愈合作用的实验研究

    Institute of Scientific and Technical Information of China (English)

    武雪芬; 张松亮; 侯益民; 孙得梅

    2012-01-01

    目的:观察水杨酸-g-壳聚糖对实验性小鼠烫伤创面愈合的影响.方法:将小鼠进行烫伤造模处理后,随机将其分为空白组、模型组、京万红组、水杨酸-g-壳聚糖组和溶剂组;通过用药前后小鼠的体征变化、皮肤溃疡疮面愈合情况及病理形态来观察和评价水杨酸-g-壳聚糖对创面的愈合作用.结果:水杨酸-g-壳聚糖对烫伤所致溃疡有明显的促进其愈合的作用,其疗效与京万红软膏相当甚至优于京万红.结论:水杨酸-g-壳聚糖可以作为抗溃疡药进一步研究.%Objective;To observe the effect of ortho - oxybenzoic acid-g-chitosan on healing of skin scald of experimental rats. Methods: The model of skin scald of rats was established, and then the rats were randomly divided into 4 groups, including blank group, model group,jingwanhong ointment group,ortho - oxybenzoic acid-g-chitosan group and solvent group. Observing and estimating the effect of healing of ortho - oxybenzoic acid-g-chitosan on scalded wound from the change of physical signs, the condition of wound healing and pathological form of rats before and after medication. Results: Ortho - oxybenzoic acid-g-chitosan has the obvious effect of accelerating wound healing on skin scald,and has the same and even better medical efficacy than jingwanhong ointment. Conclusion: Ortho - oxybenzoic acid-g-chitosan can be further studied as a antiulcer medicine.

  7. Clonal spread of catalase-negative ST5/SCCmec II Staphylococcus aureus carrying the staphylococcal enterotoxin A (sea), staphylococcal enterotoxin b (seb), and toxic shock toxin (tst) virulence genes.

    Science.gov (United States)

    Lee, Hae Kyung; Kim, Jung-Beom; Kim, Hyunjung; Jekarl, Dong Wook; Kim, Yang Ree; Yu, Jin Kyung; Park, Yeon-Joon

    2014-01-01

    17 catalase-negative methicillin-resistant Staphylococcus aureus (MRSA) isolates were recovered from respiratory specimens of patients at a 700-bed hospital in Korea. The goal of this study was to determine the molecular characteristics of catalase-negative MRSA strains in Korea for the first time. Characteristics that we explored included kat A gene mutation sequence, sequence type, staphylococcal cassette chromosome (SCC) mec subtype classification, and toxin gene profiles. All 17 isolates showed similar pulsed field gel electrophoresis (PFGE) pattern. Four mutations were identified in the kat A gene of a representative catalase-negative MRSA strain: A602G causing a histidine 201 to arginine change, A695T causing a glutamic acid 232 to valine change, T778A causing a tryptophan 260 to arginine change, and G1438A causing a glycine 480 to serine change. Previous studies suggest that the A695T and T778A mutations may have strong effects on the catalase activity of catalase-negative MRSA. The sequence type (ST) and SCCmec type of this isolate were ST 5 and SCCmec type II, respectively. All 17 isolates harbored toxic shock toxin (tst), staphylococcal enterotoxin A (sea), and staphylococcal enterotoxin B (seb) virulence genes. The mortality rate of the present study was 11.8%, suggesting that the clinical relevance of catalase-negative MRSA requires further study in the future.

  8. Characterization of the residual structure in the unfolded state of the Delta 131 Delta fragment of staphylococcal nuclease

    DEFF Research Database (Denmark)

    Francis, C. J.; Lindorff-Larsen, Kresten; Best, R. B.;

    2006-01-01

    The determination of the conformational preferences in unfolded states of proteins constitutes an important challenge in structural biology. We use inter-residue distances estimated from site-directed spin-labeling NMR experimental measurements as ensemble-averaged restraints in all-atom molecular...... dynamics simulations to characterise the residual structure of the 131 fragment of staphylococcal nuclease under physiological conditions. Our findings indicate that 131 under these conditions shows a tendency to form transiently hydrophobic clusters similar to those present in the native state of wild......-type staphylococcal nuclease. Only rarely, however, all these interactions are simultaneously realized to generate conformations with an overall native topology. Proteins 2006. © 2006 Wiley-Liss, Inc....

  9. Structural Comparison of Three Types of Staphylococcal Cassette Chromosome mec Integrated in the Chromosome in Methicillin-Resistant Staphylococcus aureus

    OpenAIRE

    Ito, Teruyo; Katayama, Yuki; Asada, Kazumi; Mori, Namiko; Tsutsumimoto, Kanae; Tiensasitorn, Chuntima; Hiramatsu, Keiichi

    2001-01-01

    The β-lactam resistance gene mecA of Staphylococcus aureus is carried by a novel mobile genetic element, designated staphylococcal cassette chromosome mec (SCCmec), identified in the chromosome of a Japanese methicillin-resistant S. aureus (MRSA) strain. We now report identification of two additional types of mecA-carrying genetic elements found in the MRSA strains isolated in other countries of the world. There were substantial differences in the size and nucleotide sequences between the ele...

  10. Staphylococcal superantigen-specific IgE antibodies: degree of sensitization and association with severity of asthma

    Science.gov (United States)

    Elabras, José; Mello, Fernanda Carvalho de Queiroz; Lupi, Omar; Bica, Blanca Elena Rios Gomes; Papi, José Angelo de Souza; França, Alfeu Tavares

    2016-01-01

    ABSTRACT Objective: To determine the presence of staphylococcal superantigen-specific IgE antibodies and degree of IgE-mediated sensitization, as well as whether or not those are associated with the severity of asthma in adult patients. Methods: This was a cross-sectional study involving outpatients with asthma under treatment at a tertiary care university hospital in the city of Rio de Janeiro, Brazil. Consecutive patients were divided into two groups according to the severity of asthma based on the Global Initiative for Asthma criteria: mild asthma (MA), comprising patients with mild intermittent or persistent asthma; and moderate or severe asthma (MSA). We determined the serum levels of staphylococcal toxin-specific IgE antibodies, comparing the results and performing a statistical analysis. Results: The study included 142 patients: 72 in the MA group (median age = 46 years; 59 females) and 70 in the MSA group (median age = 56 years; 60 females). In the sample as a whole, 62 patients (43.7%) presented positive results for staphylococcal toxin-specific IgE antibodies: staphylococcal enterotoxin A (SEA), in 29 (20.4%); SEB, in 35 (24.6%); SEC, in 33 (23.2%); and toxic shock syndrome toxin (TSST), in 45 (31.7%). The mean serum levels of IgE antibodies to SEA, SEB, SEC, and TSST were 0.96 U/L, 1.09 U/L, 1.21 U/L, and 1.18 U/L, respectively. There were no statistically significant differences between the two groups in terms of the qualitative or quantitative results. Conclusions: Serum IgE antibodies to SEA, SEB, SEC, and TSST were detected in 43.7% of the patients in our sample. However, neither the qualitative nor quantitative results showed a statistically significant association with the clinical severity of asthma. PMID:27812635

  11. [The significance of some potentially pathogenic microorganisms in occurrence of food toxicosis. Part 1. S. aureus and staphylococcal enterotoxins].

    Science.gov (United States)

    Efimochkina, N R; Kuvaeva, I B; Fluer, F S

    2011-01-01

    The data on the nomenclature, classification and properties of staphylococci and staphylococcal enterotoxins produced by them are presented. The analysis of cultural and biochemical properties of 137 strains of staphylococci isolated from raw milk and "Russian" cheese was performed. The high degree of correlation between the ability of S. aureus produce enterotoxins and the presence of enzymes coagulase, thermostable DNase, and other factors of pathogenicity is established.

  12. Expression and characterization of single-chain variable fragment antibody against staphylococcal enterotoxin A in Escherichia coli.

    Science.gov (United States)

    Chen, Weifeng; Hu, Li; Liu, Aiping; Li, Jinquan; Chen, Fusheng; Wang, Xiaohong

    2014-11-01

    The staphylococcal enterotoxins (SEs) are potent gastrointestinal exotoxins synthesized by Staphylococcus aureus, which is responsible for various diseases including septicemia, food poisoning, and toxic shock syndrome, as well as bovine mastitis. Among them, staphylococcal enterotoxin A (SEA) is one of the most commonly present serotypes in staphylococcal food poisoning cases. In this study, the stable hybridoma 3C12 producing anti-SEA monoclonal antibody was established with an equilibrium dissociation constant (KD) of 1.48 × 10(-8) mol·L(-1), its ScFv-coding genes were obtained and then the anti-SEA single chain variable fragment (ScFv) protein was expressed in Escherichia coli. Characterization of the expressed target ScFv protein was analyzed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis, Western blot, and enzyme-linked immunosorbent assay. The results demonstrated that the recombinant anti-SEA ScFv protein retained a specific binding activity for SEA, and the KD value of the soluble ScFv was about 3.75 × 10(-7) mol·L(-1). The overall yield of bioactive anti-SEA ScFv in E. coli flask culture was more than 10 mg·L(-1).

  13. Inhibition of exotoxin production by mobile genetic element SCCmec-encoded psm-mec RNA is conserved in staphylococcal species.

    Science.gov (United States)

    Ikuo, Mariko; Nagano, Gentaro; Saito, Yuki; Mao, Han; Sekimizu, Kazuhisa; Kaito, Chikara

    2014-01-01

    Staphylococcal species acquire antibiotic resistance by incorporating the mobile-genetic element SCCmec. We previously found that SCCmec-encoded psm-mec RNA suppresses exotoxin production as a regulatory RNA, and the psm-mec translation product increases biofilm formation in Staphylococcus aureus. Here, we examined whether the regulatory role of psm-mec on host bacterial virulence properties is conserved among other staphylococcal species, S. epidermidis and S. haemolyticus, both of which are important causes of nosocomial infections. In S. epidermidis, introduction of psm-mec decreased the production of cytolytic toxins called phenol-soluble modulins (PSMs) and increased biofilm formation. Introduction of psm-mec with a stop-codon mutation that did not express PSM-mec protein but did express psm-mec RNA also decreased PSM production, but did not increase biofilm formation. Thus, the psm-mec RNA inhibits PSM production, whereas the PSM-mec protein increases biofilm formation in S. epidermidis. In S. haemolyticus, introduction of psm-mec decreased PSM production, but did not affect biofilm formation. The mutated psm-mec with a stop-codon also caused the same effect. Thus, the psm-mec RNA also inhibits PSM production in S. haemolyticus. These findings suggest that the inhibitory role of psm-mec RNA on exotoxin production is conserved among staphylococcal species, although the stimulating effect of the psm-mec gene on biofilm formation is not conserved.

  14. 中药及榆烧伤止痛膏创新治疗烧烫伤78例%Treating78 cases of burns and scalds with the Jiyu Shaoshang Zhitong cream

    Institute of Scientific and Technical Information of China (English)

    于豪

    2015-01-01

    烧烫伤后,皮肤表面失去屏障极易引起细菌感染,严重时可引发败血症等恶性疾病,甚至导致死亡。烧烫伤的常规治疗方法是对症处理,如给患者口服或注射止痛剂,外用西药消炎生肌,但疗效往往不令人满意。近年来,笔者尝试采用及榆烧伤止痛膏治疗烧烫伤患者78例,疗效满意,希望有助于同道。%After the burns and scalds, the skin surface loses barrier; This situation can easily lead to bacterial infection, in some severe cases also can lead to sepsis and other malignant diseases, and even to death. Conventional treatment is symptomatic, such as oral administration or injecting analgesics, and anti-inflammatory for the skin surface with western medicine; but the effect is often unsatisfactory. In recent years, 78 cases of burns and scalds were treated with the Jiyu Shaoshang Zhitong cream; the results were satisfactory, hope to provide some suggestions for the fellows.

  15. 成都市预防儿童烧烫伤知识和行为研究分析%Survey and analysis on prevention knowledge and behavior of children scald in Chengdu

    Institute of Scientific and Technical Information of China (English)

    王艳梅; 夏劲节; 刘熹; 张晓燕

    2014-01-01

    Objective To understand the situation of prevention knowledge and behavior of family burn and scald among children in different areas in Chengdu so as to provide suggestion for decreasing the risk of family burn and scald for children. Methods In Oct. 2012, 1850 children in 12 schools were selected by group sampling and cluster sampling to finish the questionnaire survey. Results The average score of prevention knowledge about family burn and scald was 3.95±1.20(total score was 7). The score of children who living in urban areas is higher than those living in suburb(P<0.05). The proportion of family who install smoke alarms in home was low, other proportion of correct behavior was above 60%. The correct behavior proportion of children who living in outer suburb was lower than those living in other areas(P<0.05). Conclusions The prevention knowledge about family burn and scald among children in Chengdu was relatively weak. The correct behavior rate is also different in different areas. So we should focus on the key point, strengthen the intervention measures to decrease the risk of family burn and scald among children.%[目的]了解成都市不同地区儿童预防家庭烧烫伤知识和行为现况,为降低儿童家庭烧烫伤风险提供建议。[方法]2012年10月选择成都市12所学校,采用分层和整群抽样方法,对1850名3-5年级小学生进行问卷调查。[结果]儿童对家庭烧烫伤知识的平均得分为(3.95±1.20)分(总分7分),主城区儿童的平均得分比其他地区高(P<0.05)。在行为方面,屋内装有烟雾报警器的学生家庭比例较低,其他行为正确率均在60.0%以上。远郊区县儿童预防火灾的家庭安全行为正确率均低于其他地区(P<0.05)。[结论]成都市儿童对预防烧烫伤的知识相对薄弱,不同地区儿童的行为正确率也有差别,应该突出重点,加强针对性的制定干预措施,降低儿童家庭烧烫伤风险。

  16. Outbreak of staphylococcal food intoxication after consumption of pasteurized milk products, June 2007, Austria.

    Science.gov (United States)

    Schmid, Daniela; Fretz, Rainer; Winter, Petra; Mann, Michaela; Höger, Gerda; Stöger, Anna; Ruppitsch, Werner; Ladstätter, Johann; Mayer, Norbert; de Martin, Alfred; Allerberger, Franz

    2009-01-01

    On June 13, 2007, the public health authority informed the Austrian Agency for Health and Food Safety about 40 children from two neighboring elementary schools who had fallen ill with abdominal cramps and vomiting on June 8. School milk products consumed on June 8 were suspected as the source of the outbreak. On June 8, the milk products provided by local dairy X to eight elementary schools and two nurseries. The short incubation period - all cases fell ill on the day on which the products were consumed - and the short duration of illness (1-2 days) strongly suggested intoxication. In order to identify the causative pathogen, its reservoir and the mode of transmission, a descriptive-epidemiological and microbiological investigation and a retrospective cohort study were conducted. Six of the 10 institutions served by dairy X completed questionnaires on demographics and food consumption. One school had a 79% response rate (203/258) and was chosen as the basis for our cohort study. A total of 166 of the 1025 children (16.2%) at the 10 institutions fulfilled the case definition. Consumption of milk, cacao milk or vanilla milk originating from dairy X was associated with a 37.8 times higher risk of becoming a case (95% CI: 2.3-116.5). Unopened milk products left over at the affected institutions yielded staphylococcal enterotoxins A and D. Six out of 64 quarter milk samples from three of 16 cows producing milk for dairy X tested positive for S. aureus. The isolates produced enterotoxins A and D, yielded genes encoding enterotoxins and D, and showed spa type t2953. S. aureus isolated from the nasal swab of the dairy owner harbored genes encoding enterotoxins C, G, H and I, and showed spa type t635. Our investigation revealed that the milk products produced in dairy X on June 7 were the source of the outbreak on June 8. The cows - not the dairy owner - the likely reservoir of the enterotoxin-producing S. aureus. From the risk assessment of the production process at the

  17. Analisis Tipe Staphylococcal Cassette Chromosome mec (SCCmec Isolat Methicillin Resistant Staphylococcus aureus (MRSA

    Directory of Open Access Journals (Sweden)

    Sunarjati Sudigdoadi

    2010-12-01

    Full Text Available Resistance of methicillin resistant Staphylococcus aureus (MRSA were based mainly on insertion of mobile genetic elements namely Staphylococcal cassette chromosome mec (SCCmec in the chromosome of Staphylococcus aureus. SCCmec consists of recombinase genes (ccr, mec genes complex, additional resistance genes, and insertion sequences. Recombinase genes structure mediates transfer of SCCmec from one bacteria to another. Identification of SCCmec is very important to know basic genetic resistance and to predict spreading of MRSA. The aim of this research was to analyze SCCmec type and antimicrobial susceptibility patterns. The design of this study was observational analytic study by typing SCCmec and antimicrobial susceptibility testing on July– December 2007. Isolation and identification of 45 MRSA isolates was performed in the Department of Microbiology, Faculty of Medicine, University of Padjadjaran, whereas identification of mecA gene and typing of SCCmec by multiplex PCR was performed in the Department of Microbiology, Faculty of Medicine, Sriwijaya University, Palembang. The result showed that all isolates contained mecA gene. Multiplex PCR revealed that 40 MRSA isolates had SCCmec type III and 5 isolates with type IV. All SCCmec type III isolates were multiresistant and all of the type IV were not multiresistant. In conclusion, MRSA isolates with SCCmec type III was associated with multiresistant whereas type IV was not.

  18. Inactivation of staphylococcal virulence factors using a light-activated antimicrobial agent

    Directory of Open Access Journals (Sweden)

    Wilson Michael

    2009-10-01

    Full Text Available Abstract Background One of the limitations of antibiotic therapy is that even after successful killing of the infecting microorganism, virulence factors may still be present and cause significant damage to the host. Light-activated antimicrobials show potential for the treatment of topical infections; therefore if these agents can also inactivate microbial virulence factors, this would represent an advantage over conventional antibiotic therapy. Staphylococcus aureus produces a wide range of virulence factors that contribute to its success as a pathogen by facilitating colonisation and destruction of host tissues. Results In this study, the ability of the light-activated antimicrobial agent methylene blue in combination with laser light of 665 nm to inactivate staphylococcal virulence factors was assessed. A number of proteinaceous virulence factors were exposed to laser light in the presence of methylene blue and their biological activities re-determined. The activities of V8 protease, α-haemolysin and sphingomyelinase were shown to be inhibited in a dose-dependent manner by exposure to laser light in the presence of methylene blue. Conclusion These results suggest that photodynamic therapy could reduce the harmful impact of preformed virulence factors on the host.

  19. Superantigen staphylococcal enterotoxin C1 mutant can reduce paraquat pulmonary fibrosis.

    Science.gov (United States)

    Li, Tiegang; Xu, Mingkai; Wang, Nana; Zhao, Min

    2015-01-01

    A network of inflammation factors is related to pulmonary fibrosis induced by paraquat (PQ) poisoning. At high doses, the superantigen staphylococcal enterotoxin C1 (SEC1) can induce immunological unresponsiveness and inhibit release of inflammation factors. In this study, site-directed mutagenesis was performed at the H118 and H122 amino acid residues of SEC1 to reduce SEC1 toxicity. The SEC1 mutant showed significantly decreased pyrogenic toxicity, but retained clonal anergy at high dosages in vitro. Pretreatment with the SEC1 mutant prior to PQ poisoning in mice reduced symptom duration and severity, prolonged survival time, and decreased the splenocyte response to ConA induction. The SEC1 mutant also down-regulated several important cytokines related to fibrosis in the plasma after PQ poisoning. SEC1 decreased the expression of genes related to pulmonary fibrosis, including α-SMA, COL1a1, COL3 and TGF-β1, in PQ poisoned mice. Histomorphological observation indicated alleviation of pathological changes in the lungs after SEC1 pretreatment compared to mice in the PQ group. In conclusion, the SEC1 mutant reduced pulmonary interstitial fibrosis induced by PQ poisoning.

  20. Kadar Interleukin-18 pada Kultur Limfosit Penderita Dermatitis Atopik yang Distimulasi Staphylococcal Enterotoxin B (SEB

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    Oki Suwarsa

    2015-12-01

    Full Text Available Staphylococcus aureus (S. aureus has an important role in the pathogenesis of atopic dermatitis (AD. S. aureus acts as a triggering factor for AD and also causes chronic inflammation. These roles of S. aureus are related to various proteins such as Staphylococcal enterotoxin B (SEB as a potent toxin. Interleukin-18 (IL-18 is an important regulator of cytokine production of Th-1, which is interferon-γ (IFN-γ. The aim of this study was to reveal the levels of IL-18 in cultured lymphocytes from AD patients exposed by SEB. This study was conducted on 20 people with DA (7 men and 13 women and 20 healthy volunteers (9 men and 11 women in dr. Hasan Sadikin General Hospital. The in vitro experimental study on cultured lympocytes exposed with SEB was performed at the Integrated Research and Testing Laboratory of Gadjah Mada University. The average levels of IL-18 in cultured lymphocytes before and after being exposed to SEB increased both in AD group and control group. After the statistical tests was performed on the ratio of the average levels of IL-18 before and after being exposed to SEB between AD and control groups, it was shown that the levels of IL-18 AD group was significantly higher than the control group (p <0.05. Therefore, it can be concluded that the levels of IL-18 increased higher in AD group exposed by SEB.

  1. Trace Levels of Staphylococcal Enterotoxin Bioactivity Are Concealed in a Mucosal Niche during Pulmonary Inflammation.

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    Antoine Ménoret

    Full Text Available Pathogen and cellular by-products released during infection or trauma are critical for initiating mucosal inflammation. The localization of these factors, their bioactivity and natural countermeasures remain unclear. This concept was studied in mice undergoing pulmonary inflammation after Staphylococcal enterotoxin A (SEA inhalation. Highly purified bronchoalveolar lavage fluid (BALF fractions obtained by sequential chromatography were screened for bioactivity and subjected to mass spectrometry. The Inflammatory and inhibitory potentials of the identified proteins were measured using T cells assays. A potent pro-inflammatory factor was detected in BALF, and we hypothesized SEA could be recovered with its biological activity. Highly purified BALF fractions with bioactivity were subjected to mass spectrometry. SEA was the only identified protein with known inflammatory potential, and unexpectedly, it co-purified with immunosuppressive proteins. Among them was lactoferrin, which inhibited SEA and anti-CD3/-CD28 stimulation by promoting T cell death and reducing TNF synthesis. Higher doses of lactoferrin were required to inhibit effector compared to resting T cells. Inhibition relied on the continual presence of lactoferrin rather than a programming event. The data show a fraction of bioactive SEA resided in a mucosal niche within BALF even after the initiation of inflammation. These results may have clinical value in human diagnostic since traces levels of SEA can be detected using a sensitive bioassay, and may help pinpoint potential mediators of lung inflammation when molecular approaches fail.

  2. High sensitivity chemiluminescence enzyme immunoassay for detecting staphylococcal enterotoxin A in multi-matrices.

    Science.gov (United States)

    Zhang, Chunmei; Liu, Zhijia; Li, Yongming; Li, Qi; Song, Chaojun; Xu, Zhuwei; Zhang, Yun; Zhang, Yusi; Ma, Ying; Sun, Yuanjie; Chen, Lihua; Fang, Liang; Yang, Angang; Yang, Kun; Jin, Boquan

    2013-09-24

    In this study, detection of staphylococcal enterotoxin A (SEA) in multi-matrices using a highly sensitive and specific microplate chemiluminescence enzyme immunoassay (CLEIA) has been established. A pair of monoclonal antibodies (mAbs) was selected from 37 anti-SEA mAbs by pairwise analysis, and the experimental conditions of the CLEIA were optimized. This CLEIA exhibited high performance with a wide dynamic range from 6.4 pg mL(-1) to 1600 pg mL(-1), and the measured low limit of detection (LOD) was 3.2 pg mL(-1). No cross-reactivity was observed when this method was applied to test SEB, SEC1, and SED. It has also been successfully applied for analyzing SEA in a variety of environmental, biological, and clinical matrices, such as sewage, tap water, river water, roast beef, peanut butter, cured ham, 10% nonfat dry milk, milk, orange juice, human urine, and serum. Thus, the highly sensitive and SEA-specific CLEIA should make it attractive for quantifying SEA in public health and diagnosis in near future.

  3. An outbreak of staphylococcal food poisoning in the Municipality of Passos, MG, Brazil

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    Carmo Luiz Simeão do

    2003-01-01

    Full Text Available An outbreak of staphylococcal food poisoning involving 42 people who had eaten a meal at a restaurant in the Municipality of Passos, Minas Gerais, Brazil, is reported. Thirty-one of the individuals became ill with vomiting, diarrhea and dizziness within 30 minutes after eating the meal. The foods suspected were: chicken pancake, rice, beans, tomato sauce and mashed chick-peas. Large numbers (> 2.0x10(8 CFU/g of enterotoxigenic staphylococci were present in the chicken pancake. These strains produced enterotoxins A, B and D. Swabs from the nasal cavity and throat and from under the fingernails of food handlers were cultured for the detection of enterotoxigenic staphylococci carriers. Four out of five of them were healthy carriers of enterotoxin A, B, C and D producing Staphylococcus aureus at the sites cultured and one of them was also a nasal carrier of TSST-1 toxin producing S. aureus. These results indicate that the food handlers would have been the source of the food contamination.

  4. Staphylococcal SSL5 Binding to Human Leukemia Cells Inhibits Cell Adhesion to Endothelial Cells and Platelets

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    Annemiek M. E. Walenkamp

    2010-01-01

    Full Text Available Bacterial proteins provide promising tools for novel anticancer therapies. Staphylococcal superantigen-like 5 (SSL5 was recently described to bind P-selectin glycoprotein ligand-1 (PSGL-1 on leukocytes and to inhibit neutrophil rolling on a P-selectin surface. As leukocytes and tumor cells share many characteristics in migration and dissemination, we explored the potential of SSL5 as an antagonist of malignant cell behavior. Previously, it was demonstrated that rolling of human HL-60 leukemia cells on activated endothelial cells was mediated by P-selectin. In this study, we show that SSL5 targets HL-60 cells. Binding of SSL5 was rapid and without observed toxicity. Competition of SSL5 with the binding of three anti-PSGL-1 antibodies and P-selectin to HL-60 cells identified PSGL-1 as the ligand on HL-60 cells. Presence of sialyl Lewis x epitopes on PSGL-1 was crucial for its interaction with SSL5. Importantly, SSL5 not only inhibited the interaction of HL-60 cells with activated endothelial cells but also with platelets, which both play an important role in growth and metastasis of cancers. These data support the concept that SSL5 could be a lead in the search for novel strategies against hematological malignancies.

  5. Staphylococcal SCCmec elements encode an active MCM-like helicase and thus may be replicative

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    Mir-Sanchis, Ignacio [Univ. of Chicago, IL (United States). Dept. of Biochemistry and Molecular Biology; Roman, Christina A. [Univ. of Chicago, IL (United States). Dept. of Biochemistry and Molecular Biology; Misiura, Agnieszka [Univ. of Chicago, IL (United States). Dept. of Biochemistry and Molecular Biology; Abbott Lab., North Chicago, IL (United States); Pigli, Ying Z. [Univ. of Chicago, IL (United States). Dept. of Biochemistry and Molecular Biology; Boyle-Vavra, Susan [Univ. of Chicago, IL (United States). Dept. of Pediatrics; Univ. of Chicago, IL (United States). MRSA Research Center; Rice, Phoebe A. [Univ. of Chicago, IL (United States). Dept. of Biochemistry and Molecular Biology

    2016-08-29

    Methicillin-resistant Staphylococcus aureus (MRSA) is a public-health threat worldwide. Although the mobile genomic island responsible for this phenotype, staphylococcal cassette chromosome (SCC), has been thought to be nonreplicative, we predicted DNA-replication-related functions for some of the conserved proteins encoded by SCC. We show that one of these, Cch, is homologous to the self-loading initiator helicases of an unrelated family of genomic islands, that it is an active 3'-to-5' helicase and that the adjacent ORF encodes a single-stranded DNA–binding protein. Our 2.9-Å crystal structure of intact Cch shows that it forms a hexameric ring. Cch, like the archaeal and eukaryotic MCM-family replicative helicases, belongs to the pre–sensor II insert clade of AAA+ ATPases. Additionally, we found that SCC elements are part of a broader family of mobile elements, all of which encode a replication initiator upstream of their recombinases. Replication after excision would enhance the efficiency of horizontal gene transfer.

  6. Intranasal Rapamycin Rescues Mice from Staphylococcal Enterotoxin B-Induced Shock

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    Teresa Krakauer

    2012-09-01

    Full Text Available Staphylococcal enterotoxin B (SEB and related exotoxins produced by Staphylococcus aureus are potent activators of the immune system and cause toxic shock in humans. Currently there is no effective treatment except for the use of intravenous immunoglobulins administered shortly after SEB exposure. Intranasal SEB induces long-lasting lung injury which requires prolonged drug treatment. We investigated the effects of rapamycin, an immunosuppressive drug used to prevent graft rejection, by intranasal administration in a lethal mouse model of SEB-induced shock. The results show that intranasal rapamycin alone delivered as late as 17 h after SEB protected 100% of mice from lethal shock. Additionally, rapamycin diminished the weight loss and temperature fluctuations elicited by SEB. Intranasal rapamycin attenuated lung MCP-1, IL-2, IL-6, and IFNγ by 70%, 30%, 64%, and 68% respectively. Furthermore, short courses (three doses of rapamycin were sufficient to block SEB-induced shock. Intranasal rapamycin represents a novel use of an immunosuppressant targeting directly to site of toxin exposure, reducing dosages needed and allowing a wider therapeutic window.

  7. Evaluation of Handheld Assays for the Detection of Ricin and Staphylococcal Enterotoxin B in Disinfected Waters

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    Mary Margaret Wade

    2011-01-01

    Full Text Available Development of a rapid field test is needed capable of determining if field supplies of water are safe to drink by the warfighter during a military operation. The present study sought to assess the effectiveness of handheld assays (HHAs in detecting ricin and Staphylococcal Enterotoxin B (SEB in water. Performance of HHAs was evaluated in formulated tap water with and without chlorine, reverse osmosis water (RO with chlorine, and RO with bromine. Each matrix was prepared, spiked with ricin or SEB at multiple concentrations, and then loaded onto HHAs. HHAs were allowed to develop and then read visually. Limits of detection (LOD were determined for all HHAs in each water type. Both ricin and SEB were detected by HHAs in formulated tap water at or below the suggested health effect levels of 455 ng/mL and 4.55 ng/mL, respectively. However, in brominated or chlorinated waters, LODs for SEB increased to approximately 2,500 ng/mL. LODs for ricin increased in chlorinated water, but still remained below the suggested health effect level. In brominated water, the LOD for ricin increased to approximately 2,500 ng/mL. In conclusion, the HHAs tested were less effective at detecting ricin and SEB in disinfected water, as currently configured.

  8. Mupirocin in the Treatment of Staphylococcal Infections in Chronic Rhinosinusitis: A Meta-Analysis

    Science.gov (United States)

    2016-01-01

    Background Saline irrigation of the nasal cavity is a classic and effective treatment for acute or chronic rhinosinusitis. Topical antibiotics such as mupirocin have been widely used for recalcitrant chronic rhinosinusitis. Therefore, the purpose of this study was to evaluate the effect of saline irrigation using mupirocin. Methods A systematic literature review and meta-analysis of mupirocin saline irrigation were performed using EMBASE, MEDLINE, and Cochrane library through December 2015. Data were analyzed with R 3.2.2 software. A random effects model was used because of the diversity of included studies. Sensitivity analysis of particular tested groups and single proportion tests were also performed. The main outcome measure was residual staphylococcal infection, as confirmed by culture or PCR. Results Two RCTs, two prospective studies and two retrospective studies were included. A random effects model meta-analysis of the pooled data identified a relative risk of residual infection of 0.13 (95% CI: 0.06–0.26, prhinosinusitis. Although there is currently a lack of clear evidence, future studies with well-designed inclusion criteria and randomized controlled trials are needed to examine mupirocin’s long-term effect on chronic rhinosinusitis. PMID:27907108

  9. Effect and Mechanism of Superantigen Staphylococcal Enterotoxin Therapy for Mouse Gastric Tumor

    Institute of Scientific and Technical Information of China (English)

    李毅清; 舒晓钢

    2003-01-01

    Summary.. The anti-tumor effect and mechanism of the staphylococcal enterotoxin A (SEA) werestudied. The mouse gastric tumor model was produced by subcutaneously inoculating gastric tumorcells (MGC80-3). The experimental group was treated with SEA, and the control group was treatedwith normal saline. The percentage of tumor generation and tumor mass was measured. The resultsshowed that the percentage of the tumor generation in the SEA-treated mice was lower than in thecontrol group, but there was no significant difference (P>0. 05). However, the tumor mass in theexperimental group was significantly lighter than in the control group, with the difference being verysignificant (P<0. 001). There were more CD+4 T cells and CD+8 T cells in the tumor of the micetreated with SEA than those of the control group. SEA has an obvious anti-tumor effect on mice gas-tric tumor. The mechanism might be that SEA induces the effect of superantigen-dependent cell me-diated cytotoxicity to the tumor cells.

  10. Inhibition of the anti-staphylococcal activity of the antiseptic polihexanide by mucin.

    Science.gov (United States)

    Ansorg, Rainer; Rath, Peter-Michael; Fabry, Werner

    2003-01-01

    The antiseptic Lavasept (LS), containing the polymeric biguanide polihexanide (CAS 28757-48-4), possesses microbicidal activity against a broad spectrum of bacteria including Staphylococcus aureus. It is used for antiseptic wound care in concentrations corresponding to 0.2-0.4 mg polihexanide per ml. To obtain basic data on its ability to eradicate S. aureus colonizing the nasal mucosa, the influence of mucin on the anti-staphylococcal activity was investigated. A disk agar-diffusion method was applied. Two reference strains of S. aureus (methicillin-sensitive S. aureus ATCC 29213 and methicillin-resistant S. aureus ATCC 33591) and 20 fresh clinical isolates were used. In the absence of mucin, the growth of all strains was inhibited by polihexanide concentrations of 0.1 mg/ml. In the presence of 0.25% mucin in the test medium, a concentration of 0.4 mg/ml was necessary to inhibit all strains. Mucin concentrations of 0.5% and 1%, that are even lower than the mucin concentrations in healthy nasal secretions, abolished the activity of the therapeutic concentrations of polihexanide. It is concluded that the inactivation of LS by mucin obstructs a reliable clearance of nasal S. aureus carriage.

  11. Variation of serine-aspartate repeats in membrane proteins possibly contributes to staphylococcal microevolution.

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    Jing Cheng

    Full Text Available Tandem repeats (either as microsatellites or minisatellites in eukaryotic and prokaryotic organisms are mutation-prone DNA. While minisatellites in prokaryotic genomes are underrepresented, the cell surface adhesins of bacteria often contain the minisatellite SD repeats, encoding the amino acid pair of serine-asparatate, especially in Staphylococcal strains. However, their relationship to biological functions is still elusive. In this study, effort was made to uncover the copy number variations of SD repeats by bioinformatic analysis and to detect changes in SD repeats during a plasmid-based assay, as a first step to understand its biological functions. The SD repeats were found to be mainly present in the cell surface proteins. The SD repeats were genetically unstable and polymorphic in terms of copy numbers and sequence compositions. Unlike SNPs, the change of its copy number was reversible, without frame shifting. More significantly, a rearrangement hot spot, the ATTC/AGRT site, was found to be mainly responsible for the instability and reversibility of SD repeats. These characteristics of SD repeats may facilitate bacteria to respond to environmental changes, with low cost, low risk and high efficiency.

  12. Outbreak of staphylococcal food poisoning from a military unit lunch party - United States, July 2012.

    Science.gov (United States)

    2013-12-20

    On July 30, 2012, the emergency department at a military hospital was visited by 13 persons seeking care for gastrointestinal illness with onset 2-3 hours after a work lunch party. The hospital responded by opening up temporary evaluation and treatment capacity in primary-care clinics and a progressive-care unit and by diverting one patient to a local civilian hospital. An immediate outbreak investigation was conducted by local military public health personnel with assistance from CDC. Initial epidemiologic analysis implicated "perlo" (a chicken, sausage, and rice dish) and bacterial intoxication as the outbreak mechanism. This enabled public health personnel to 1) recommend no further consumption of perlo and 2) reassure appropriate authorities that no additional ill persons likely would be seeking care and advise that nothing more than supportive care of ill persons likely would be required. After interviewing party attendees, investigators found nine additional persons who met their case definition. Subsequent CDC laboratory analysis of a sample of perlo detected staphylococcal enterotoxin A, supporting the epidemiologic findings. Improper food handling and preparation measures were identified and addressed by the appropriate authorities, who provided additional detailed education on food preparation safety for the persons who prepared the meal.

  13. Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice.

    Science.gov (United States)

    Kerr, D E; Plaut, K; Bramley, A J; Williamson, C M; Lax, A J; Moore, K; Wells, K D; Wall, R J

    2001-01-01

    Infection of the mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry. Staphylococcus aureus is the major contagious mastitis pathogen, accounting for approximately 15-30% of infections, and has proved difficult to control using standard management practices. As a first step toward enhancing mastitis resistance of dairy animals, we report the generation of transgenic mice that secrete a potent anti-staphylococcal protein into milk. The protein, lysostaphin, is a peptidoglycan hydrolase normally produced by Staphylococcus simulans. When the native form is secreted by transfected eukaryotic cells it becomes glycosylated and inactive. However, removal of two glycosylation motifs through engineering asparagine to glutamine codon substitutions enables secretion of Gln(125,232)-lysostaphin, a bioactive variant. Three lines of transgenic mice, in which the 5'-flanking region of the ovine beta-lactoglobulin gene directed the secretion of Gln(125,232)-lysostaphin into milk, exhibit substantial resistance to an intramammary challenge of 104 colony-forming units (c.f.u.) of S. aureus, with the highest expressing line being completely resistant. Milk protein content and profiles of transgenic and nontransgenic mice are similar. These results clearly demonstrate the potential of genetic engineering to combat the most prevalent disease of dairy cattle.

  14. Effect of thermal processing during yogurt production upon the detection of staphylococcal enterotoxin B.

    Science.gov (United States)

    Principato, Maryann; Boyle, Thomas; Njoroge, Joyce; Jones, Robert L; O'Donnell, Michael

    2009-10-01

    This research was conducted to examine the inherent properties of yogurt contaminated with staphylococcal enterotoxin B (SEB). Two types of yogurts were produced for this study. Type I yogurts were produced by adding SEB at the start of yogurt production, and type II yogurts were produced by adding SEB after the milk base had been boiled. Biochemical characteristics inherent to yogurt, including pH, lactic acid and acetaldehyde concentrations, were analyzed weekly for each batch beginning at a time just after production and throughout a storage period of at least 4 weeks. The presence of toxin during yogurt production did not result in any significant biochemical or physical changes in yogurt. However, we were unable to detect SEB toxin in type I yogurt using a commercially available enzyme-linked immunosorbent assay (ELISA). In contrast, SEB was easily detectable by our ELISA in type II yogurt samples. Higher levels of SEB were recovered from type II yogurt that had been stored for 1 week than from type II yogurt that had been stored for any other length of time. These results indicate that the biochemical characteristics of yogurt did not change significantly (relative to control yogurt) in the presence of either thermally processed SEB or native SEB. However, the ability to detect SEB by ELISA was dependent on whether the toxin had been processed.

  15. Distinction between Pore Assembly by Staphylococcal α-Toxin versus Leukotoxins

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    Olivier Joubert

    2007-01-01

    Full Text Available The staphylococcal bipartite leukotoxins and the homoheptameric α-toxin belong to the same family of β-barrel pore-forming toxins despite slight differences. In the α-toxin pore, the N-terminal extremity of each protomer interacts as a deployed latch with two consecutive protomers in the vicinity of the pore lumen. N-terminal extremities of leukotoxins as seen in their three-dimensional structures are heterogeneous in length and take part in the β-sandwich core of soluble monomers. Hence, the interaction of these N-terminal extremities within structures of adjacent monomers is questionable. We show here that modifications of their N-termini by two different processes, using fusion with glutathione S-transferase (GST and bridging of the N-terminal extremity to the adjacent β-sheet via disulphide bridges, are not deleterious for biological activity. Therefore, bipartite leukotoxins do not need a large extension of their N-terminal extremities to form functional pores, thus illustrating a microheterogeneity of the structural organizations between bipartite leukotoxins and α-toxin.

  16. Epidemiological investigation of nosocomial outbreak of staphylococcal skin diseases in neonatal ward.

    Science.gov (United States)

    Kurlenda, J; Grinholc, M; Krzysztoń-Russjan, J; Wiśniewska, K

    2009-05-01

    During a 1-month period, eight neonates developed staphylococcal skin disease diagnosed as a bullous impetigo in the maternity unit of the Provincial Hospital in Gdansk. An epidemiological investigation based on phenotyping and genotyping methods was performed. All neonates involved in the outbreak, their mothers and 15 staff members were screened for carriage of Staphylococcus aureus by nasal swabs. Isolated strains were compared with strains cultured from affected skin and purulent conjunctiva of infected newborns. Isolates were analyzed for the presence of the etA and etB genes using polymerase chain reaction and genotyped by pulsed-field gel electrophoresis (PFGE) and coa gene polymorphism. The analyzed S. aureus strains were methicillin-sensitive and could be divided into two groups according to antibiotyping, phage typing, coa polymorphism and PFGE pattern. The first group consisted of etA and etB negative strains, and the second one involved only the etB positive ones. Our results have shown that there were two different clusters of infection caused by two populations of S. aureus strains. Among the 15 medical staff members screened we have found seven carriers. However, phage typing revealed that distinct strains unrelated to the outbreak isolates were carried. Although we have not been able to establish the source of bacteria involved in the outbreak, our results suggest that for both groups, mothers could be the source of the infecting strains.

  17. Staphylococcal pathogenicity island interference with helper phage reproduction is a paradigm of molecular parasitism.

    Science.gov (United States)

    Ram, Geeta; Chen, John; Kumar, Krishan; Ross, Hope F; Ubeda, Carles; Damle, Priyadarshan K; Lane, Kristin D; Penadés, José R; Christie, Gail E; Novick, Richard P

    2012-10-02

    Staphylococcal pathogenicity islands (SaPIs) carry superantigen and resistance genes and are extremely widespread in Staphylococcus aureus and in other Gram-positive bacteria. SaPIs represent a major source of intrageneric horizontal gene transfer and a stealth conduit for intergeneric gene transfer; they are phage satellites that exploit the life cycle of their temperate helper phages with elegant precision to enable their rapid replication and promiscuous spread. SaPIs also interfere with helper phage reproduction, blocking plaque formation, sharply reducing burst size and enhancing the survival of host cells following phage infection. Here, we show that SaPIs use several different strategies for phage interference, presumably the result of convergent evolution. One strategy, not described previously in the bacteriophage microcosm, involves a SaPI-encoded protein that directly and specifically interferes with phage DNA packaging by blocking the phage terminase small subunit. Another strategy involves interference with phage reproduction by diversion of the vast majority of virion proteins to the formation of SaPI-specific small infectious particles. Several SaPIs use both of these strategies, and at least one uses neither but possesses a third. Our studies illuminate a key feature of the evolutionary strategy of these mobile genetic elements, in addition to their carriage of important genes-interference with helper phage reproduction, which could ensure their transferability and long-term persistence.

  18. Application of LC-MS/MS MRM to Determine Staphylococcal Enterotoxins (SEB and SEA) in Milk.

    Science.gov (United States)

    Andjelkovic, Mirjana; Tsilia, Varvara; Rajkovic, Andreja; De Cremer, Koen; Van Loco, Joris

    2016-04-20

    Staphylococcus aureus is one of the important aetiological agents of food intoxications in Europe and can cause gastro-enteritis through the production of various staphylococcal enterotoxins (SEs) in foods. Due to their stability and ease of production and dissemination, some SEs have also been studied as potential agents for bioterrorism. Therefore, specific and accurate analytical tools are required to detect and quantify SEs. Online solid-phase extraction liquid chromatography electrospray ionization tandem mass spectrometry (online SPE-LC-ESI-MS/MS) based on multiple reaction monitoring (MRM) was used to detect and quantify two types of SE (A and B) spiked in milk and buffer solution. SE extraction and concentration was performed according to the European Screening Method developed by the European Reference Laboratory for Coagulase Positive Staphylococci. Trypsin digests were screened for the presence of SEs using selected proteotypic heavy-labeled peptides as internal standards. SEA and SEB were successfully detected in milk samples using LC-MS/MS in MRM mode. The selected SE peptides were proteotypic for each toxin, allowing the discrimination of SEA and SEB in a single run. The detection limit of SEA and SEB was approximately 8 and 4 ng/g, respectively.

  19. Assessment of the Functional Regions of the Superantigen Staphylococcal Enterotoxin B

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    Thomas J. Rogers

    2013-10-01

    Full Text Available The functional activity of superantigens is based on capacity of these microbial proteins to bind to both the β-chain of the T cell receptor (TcR and the major histocompatibility complex (MHC class II dimer. We have previously shown that a subset of the bacterial superantigens also binds to a membrane protein, designated p85, which is expressed by renal epithelial cells. This binding activity is a property of SEB, SEC1, 2 and 3, but not SEA, SED, SEE or TSST. The crystal structure of the tri-molecular complex of the superantigen staphylococcal enterotoxin B (SEB with both the TcR and class II has previously been reported. However, the relative contributions of regions of the superantigen to the overall functional activity of this superantigen remain undefined. In an effort to better define the molecular basis for the interaction of SEB with the TcR β-chain, we report studies here which show the comparative contributions of amino- and carboxy-terminal regions in the superantigen activity of SEB. Recombinant fusion proteins composed of bacterial maltose-binding protein linked to either full-length or truncated toxins in which the 81 N-terminal, or 19 or 34 C-terminal amino acids were deleted, were generated for these studies. This approach provides a determination of the relative strength of the functional activity of the various regions of the superantigen protein.

  20. Staphylococcal lipoteichoic acid promotes osteogenic differentiation of mouse mesenchymal stem cells by increasing autophagic activity.

    Science.gov (United States)

    Liu, Xin; Wang, Yuan; Cao, Zhen; Dou, Ce; Bai, Yun; Liu, Chuan; Dong, Shiwu; Fei, Jun

    2017-02-16

    This study sought to explore the effect of staphylococcal lipoteichoic acid (LTA) on autophagy in mouse mesenchymal stem cells (MSCs), and then influence osteogenesis through the change of autophagy. C3H10T1/2 cells were induced by osteogenic medium with the treatment of LTA at different concentrations (1, 5, 10 μg/mL); 3-methyladenine (3-MA) were used as the autophagy inhibitor, and rapamycin (rapamycin, Rap) were used to activate autophagy; the effects on osteogenesis were detected by alkaline phosphatase staining, alizarin red staining, real-time quantitative PCR, and western blotting; autophagic activity was investigated by the expression of LC3-Ⅱand p62 proteins. Compared with control group, the expression of osteogenesis markers was significantly up-regulated with the LTA treatment on the mRNA and protein level; the positive rate of alkaline phosphatase was enhanced in the LTA groups; and the formation of calcium nodules was increased simultaneously. The expression of LC3-Ⅱ protein was increased in LTA groups, while the expression of p62 protein was decreased. Inhibition of autophagy significantly reduced the effect of LTA on osteogenesis of MSCs; the promotion of LTA on osteogenic differentiation was further enhanced when adding rapamycin to activate autophagic activity. It provides new insight of prevention and treatment for bone infection.

  1. Staphylococcal SplB serine protease utilizes a novel molecular mechanism of activation.

    Science.gov (United States)

    Pustelny, Katarzyna; Zdzalik, Michal; Stach, Natalia; Stec-Niemczyk, Justyna; Cichon, Przemyslaw; Czarna, Anna; Popowicz, Grzegorz; Mak, Pawel; Drag, Marcin; Salvesen, Guy S; Wladyka, Benedykt; Potempa, Jan; Dubin, Adam; Dubin, Grzegorz

    2014-05-30

    Staphylococcal SplB protease belongs to the chymotrypsin family. Chymotrypsin zymogen is activated by proteolytic processing at the N terminus, resulting in significant structural rearrangement at the active site. Here, we demonstrate that the molecular mechanism of SplB protease activation differs significantly and we characterize the novel mechanism in detail. Using peptide and protein substrates we show that the native signal peptide, or any N-terminal extension, has an inhibitory effect on SplB. Only precise N-terminal processing releases the full proteolytic activity of the wild type analogously to chymotrypsin. However, comparison of the crystal structures of mature SplB and a zymogen mimic show no rearrangement at the active site whatsoever. Instead, only the formation of a unique hydrogen bond network, distant form the active site, by the new N-terminal glutamic acid of mature SplB is observed. The importance of this network and influence of particular hydrogen bond interactions at the N terminus on the catalytic process is demonstrated by evaluating the kinetics of a series of mutants. The results allow us to propose a consistent model where changes in the overall protein dynamics rather than structural rearrangement of the active site are involved in the activation process.

  2. [Description of a staphylococcal alimentary poisoning outbreak in Las Rosas, Santa Fe Province, Argentina].

    Science.gov (United States)

    Brizzio, Aníbal A; Tedeschi, Fabián A; Zalazar, Fabián E

    2011-01-01

    On February 2008, a suspected foodborne outbreak was reported in Las Rosas (Santa Fe Province, Argentina). The formal procedures indicated that an undetermined number of individuals had experienced food poisoning following consumption of vegetable cannelloni bought at a local shop. The manufacturer establishment was audited. Samples from the suspected food, as well as environmental samples and swabs from food handlers were obtained and involved subjects were interviewed. Remnants of ingested food were also obtained. Routine microbiological procedures of the foodborne outbreak revealed the presence of coagulase positive S. aureus subspecies aureus in samples from ingested and raw food, and from manipulators. Indicator microorganisms did not show significant levels and no other foodborne pathogen was isolated. Presence of staphylococcal enterotoxin-producing genes was subsequently investigated, and a positive result for enterotoxin B was shown in S. aureus strains isolated from a food handler as well as from food linked to the outbreak Moreover, these isolates showed 100% similarity by SmaI-PFGE. Timely notification together with coordinated sanitary measures and the availability of appropriate laboratory tools allowed to interrupt the chain of disease transmission by identifying risk and protective factors.

  3. Distinction between pore assembly by staphylococcal alpha-toxin versus leukotoxins.

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    Joubert, Olivier; Voegelin, Joëlle; Guillet, Valérie; Tranier, Samuel; Werner, Sandra; Colin, Didier A; Dalla Serra, Mauro; Keller, Daniel; Monteil, Henri; Mourey, Lionel; Prévost, Gilles

    2007-01-01

    The staphylococcal bipartite leukotoxins and the homoheptameric alpha-toxin belong to the same family of beta-barrel pore-forming toxins despite slight differences. In the alpha-toxin pore, the N-terminal extremity of each protomer interacts as a deployed latch with two consecutive protomers in the vicinity of the pore lumen. N-terminal extremities of leukotoxins as seen in their three-dimensional structures are heterogeneous in length and take part in the beta-sandwich core of soluble monomers. Hence, the interaction of these N-terminal extremities within structures of adjacent monomers is questionable. We show here that modifications of their N-termini by two different processes, using fusion with glutathione S-transferase (GST) and bridging of the N-terminal extremity to the adjacent beta-sheet via disulphide bridges, are not deleterious for biological activity. Therefore, bipartite leukotoxins do not need a large extension of their N-terminal extremities to form functional pores, thus illustrating a microheterogeneity of the structural organizations between bipartite leukotoxins and alpha-toxin.

  4. Identification of the blood coagulation factor interacting sequences in staphylococcal superantigen-like protein 10.

    Science.gov (United States)

    Itoh, Saotomo; Takii, Takemasa; Onozaki, Kikuo; Tsuji, Tsutomu; Hida, Shigeaki

    2017-03-25

    Staphylococcal superantigen-like proteins (SSLs) are a family of exoproteins of Staphylococcus aureus. We have shown that SSL10 binds to vitamin K-dependent coagulation factors and inhibits blood coagulation induced by recalcification of citrated plasma. SSL10 was revealed to bind to coagulation factors via their γ-carboxyglutamic acid (Gla) domain. In this study we attempted to identify the responsible sequence of SSL10 for the interaction with coagulation factors. We prepared a series of domain swap mutants between SSL10 and its paralog SSL7 that does not interact with coagulation factors, and examined their binding activity to immobilized prothrombin using ELISA-like binding assay. The domain swap mutants that contained SSL10β1-β3 ((23)MEMKN ISALK HGKNN LRFKF RGIKI QVL(60)) bound to immobilized prothrombin, and mutants that contained SSL10β10-β12 ((174)SFYNL DLRSK LKFKY MGEVI ESKQI KDIEV NLK(207)) also retained the binding activity. On the other hand, mutants that lacked these two regions did not bind to prothrombin. These sequences, each alone, bound to prothrombin as 33 amino acid length polypeptides. These results suggest that SSL10 has two responsible sequences for the binding to prothrombin. These prothrombin-binding peptides would contribute to the development of new anticoagulants.

  5. Mobile and immobile adhesion of staphylococcal strains to hydrophilic and hydrophobic surfaces.

    Science.gov (United States)

    Boks, Niels P; Kaper, Hans J; Norde, Willem; van der Mei, Henny C; Busscher, Henk J

    2009-03-01

    Staphylococcus epidermidis adheres to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in similar numbers, but in different modes. Real-time observation of staphylococcal adhesion under a shear rate of 15 s(-1) revealed different adhesion dynamics on both substrata. The number of adsorption and desorption events to achieve a similar number of adhering bacteria was twofold higher on hydrophilic than on hydrophobic DDS-coated glass. Moreover, 22% of all staphylococci on glass slid over the surface prior to adhering on a fixed site ("mobile adhesion mode"), but mobile adhesion was virtually absent (1%) on DDS-coated glass. Sliding preceded desorption on hydrophilic glass in about 20% of all desorption events, while on hydrophobic DDS-coated glass 2% of all staphylococci desorbed straight from their adhesion site. Since acid-base interactions between the staphylococci and a hydrophobic DDS-coating are attractive, it is suggested that these interactions facilitate a closer approach of the bacteria and therewith enhance immobile adhesion at local, high affinity sites. Alternatively, if the local site is low affinity, this may lead to desorption. In the absence of attractive acid-base interactions, as on hydrophilic glass, bacteria can be captured in the minimum of the DLVO-interaction energy curve, but this does not prevent them from sliding under flow at a fixed distance from a substratum surface until immobilization or desorption at or from a local high or low affinity site, respectively.

  6. ANTIBACTERIAL ACTIVITY OF Lactobacillus casei IMV B-7280 IN CASES OF EXPERIMENTAL UROGENITAL STAPHYLOCOCCAL INFE

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    Babenko L. P.

    2015-08-01

    Full Text Available The aim of work was to determine antibacterial activity of Lactobacillus casei ІМV В-7280 probiotic strain on the experimental urogenital tract infection of mice. The influence of intravaginal and/or per os administration of this strain once per day during 7 days on the microflora of vagina, kidneys and intestinal contents of Staphylococcus aureus 8325-4 infected mice was studied. It was established, that in cases of experimental staphylococcal infection of urogenital tract L. Casei IMV B-7280 had effective antagonistic activity against S. aureus 8325-4 and opportunistic bacteria. After L. casei IMV B-7280 introduction into infected mice reduction or complete elimination of S. aureus 8325-4 in vagina, kidneys and intestinal contents in different periods of observation was established. Under the influence of L. casei ІМВ В-7280 the number of coliform bacteria, streptococci and staphylococci in the vagina was normalized, and fungal flora — decreased even in comparison with intact mice. Normalization of kidneys microflora was also observed. In the intestinal contents of infected mice trea ted with L. casei IMV B-7280 the number of streptococci did not change, staphylococci number decreased, but the number of fungal and coliform flora remained relatively low during the observation period. L. casei IMV B-7280 probiotic strain is promising to create immunobiotics with antibacterial action, which can be used for the prevention and treatment of urogenital infections caused by opportunistic microorganisms.

  7. Staphylococcal Enterotoxin P Predicts Bacteremia in Hospitalized Patients Colonized With Methicillin-Resistant Staphylococcus aureus

    Science.gov (United States)

    Calderwood, Michael S.; Desjardins, Christopher A.; Sakoulas, George; Nicol, Robert; DuBois, Andrea; Delaney, Mary L.; Kleinman, Ken; Cosimi, Lisa A.; Feldgarden, Michael; Onderdonk, Andrew B.; Birren, Bruce W.; Platt, Richard; Huang, Susan S.

    2014-01-01

    Background. Methicillin-resistant Staphylococcus aureus (MRSA) colonization predicts later infection, with both host and pathogen determinants of invasive disease. Methods. This nested case-control study evaluates predictors of MRSA bacteremia in an 8–intensive care unit (ICU) prospective adult cohort from 1 September 2003 through 30 April 2005 with active MRSA surveillance and collection of ICU, post-ICU, and readmission MRSA isolates. We selected MRSA carriers who did (cases) and those who did not (controls) develop MRSA bacteremia. Generating assembled genome sequences, we evaluated 30 MRSA genes potentially associated with virulence and invasion. Using multivariable Cox proportional hazards regression, we assessed the association of these genes with MRSA bacteremia, controlling for host risk factors. Results. We collected 1578 MRSA isolates from 520 patients. We analyzed host and pathogen factors for 33 cases and 121 controls. Predictors of MRSA bacteremia included a diagnosis of cancer, presence of a central venous catheter, hyperglycemia (glucose level, >200 mg/dL), and infection with a MRSA strain carrying the gene for staphylococcal enterotoxin P (sep). Receipt of an anti-MRSA medication had a significant protective effect. Conclusions. In an analysis controlling for host factors, colonization with MRSA carrying sep increased the risk of MRSA bacteremia. Identification of risk-adjusted genetic determinants of virulence may help to improve prediction of invasive disease and suggest new targets for therapeutic intervention. PMID:24041793

  8. Review Over a 3-Year Period of European Union Proficiency Tests for Detection of Staphylococcal Enterotoxins in Food Matrices.

    Science.gov (United States)

    Nia, Yacine; Mutel, Isabelle; Assere, Adrien; Lombard, Bertrand; Auvray, Frederic; Hennekinne, Jacques-Antoine

    2016-04-13

    Staphylococcal food poisoning outbreaks are a major cause of foodborne illnesses in Europe and their notifications have been mandatory since 2005. Even though the European regulation on microbiological criteria for food defines a criterion on staphylococcal enterotoxin (SE) only in cheese and dairy products, European Food Safety Authority (EFSA) data reported that various types of food matrices are involved in staphylococcal food poisoning outbreaks. The European Screening Method (ESM) of European Union Reference Laboratory for Coagulase Positive Staphylococci (EURL CPS) was validated in 2011 for SE detection in food matrices and is currently the official method used for screening purposes in Europe. In this context, EURLCPS is annually organizing Inter-Laboratory Proficiency Testing Trials (ILPT) to evaluate the competency of the European countries' National Reference Laboratories (NRLs) to analyse SE content in food matrices. A total of 31 NRLs representing 93% of European countries participated in these ILPTs. Eight food matrices were used for ILPT over the period 2013-2015, including cheese, freeze-dried cheese, tuna, mackerel, roasted chicken, ready-to-eat food, milk, and pastry. Food samples were spiked with four SE types (i.e., SEA, SEC, SED, and SEE) at various concentrations. Homogeneity and stability studies showed that ILPT samples were both homogeneous and stable. The analysis of results obtained by participants for a total of 155 blank and 620 contaminated samples allowed for evaluation of trueness (>98%) and specificity (100%) of ESM. Further to the validation study of ESM carried out in 2011, these three ILPTs allowed for the assessment of the proficiency of the NRL network and the performance of ESM on a large variety of food matrices and samples. The ILPT design presented here will be helpful for the organization of ILPT on SE detection by NRLs or other expert laboratories.

  9. Characterization of the Staphylococcal enterotoxin A: Vβ receptor interaction using human receptor fragments engineered for high affinity.

    Science.gov (United States)

    Sharma, P; Postel, S; Sundberg, E J; Kranz, D M

    2013-12-01

    Staphylococcal food poisoning is a gastrointestinal disorder caused by the consumption of food containing Staphylococcal enterotoxins. Staphylococcal enterotoxin A (SEA) is the most common enterotoxin recovered from food poisoning outbreaks in the USA. In addition to its enteric activity, SEA also acts as a potent superantigen through stimulation of T cells, although less is known about its interactions than the superantigens SEB, SEC and toxic shock syndrome toxin-1. To understand more about SEA:receptor interactions, and to develop toxin-detection systems for use in food testing, we engineered various SEA-binding receptor mutants. The extracellular domain of the receptor, a variable region of the beta chain (Vβ22) of the T-cell receptor, was engineered for stability as a soluble protein and for high affinity, using yeast-display technology. The highest affinity mutant was shown to bind SEA with a Kd value of 4 nM. This was a 25 000-fold improvement in affinity compared with the wild-type receptor, which bound to SEA with low affinity (Kd value of 100 µM), similar to other superantigen:Vβ interactions. The SEA:Vβ interface was centered around residues within the complementarity determining region 2 loop. The engineered receptor was specific for SEA, in that it did not bind to two other closely related enterotoxins SEE or SED, providing information on the SEA residues possibly involved in the interaction. The specificity and affinity of these high-affinity Vβ proteins also provide useful agents for the design of more sensitive and specific systems for SEA detection.

  10. Review Over a 3-Year Period of European Union Proficiency Tests for Detection of Staphylococcal Enterotoxins in Food Matrices

    Directory of Open Access Journals (Sweden)

    Yacine Nia

    2016-04-01

    Full Text Available Staphylococcal food poisoning outbreaks are a major cause of foodborne illnesses in Europe and their notifications have been mandatory since 2005. Even though the European regulation on microbiological criteria for food defines a criterion on staphylococcal enterotoxin (SE only in cheese and dairy products, European Food Safety Authority (EFSA data reported that various types of food matrices are involved in staphylococcal food poisoning outbreaks. The European Screening Method (ESM of European Union Reference Laboratory for Coagulase Positive Staphylococci (EURL CPS was validated in 2011 for SE detection in food matrices and is currently the official method used for screening purposes in Europe. In this context, EURLCPS is annually organizing Inter-Laboratory Proficiency Testing Trials (ILPT to evaluate the competency of the European countries’ National Reference Laboratories (NRLs to analyse SE content in food matrices. A total of 31 NRLs representing 93% of European countries participated in these ILPTs. Eight food matrices were used for ILPT over the period 2013–2015, including cheese, freeze-dried cheese, tuna, mackerel, roasted chicken, ready-to-eat food, milk, and pastry. Food samples were spiked with four SE types (i.e., SEA, SEC, SED, and SEE at various concentrations. Homogeneity and stability studies showed that ILPT samples were both homogeneous and stable. The analysis of results obtained by participants for a total of 155 blank and 620 contaminated samples allowed for evaluation of trueness (>98% and specificity (100% of ESM. Further to the validation study of ESM carried out in 2011, these three ILPTs allowed for the assessment of the proficiency of the NRL network and the performance of ESM on a large variety of food matrices and samples. The ILPT design presented here will be helpful for the organization of ILPT on SE detection by NRLs or other expert laboratories.

  11. Association of Specific Immunoglobulin E to Staphylococcal Enterotoxin with Airway Hyperresponsiveness in Asthma Patients

    Science.gov (United States)

    Kim, Seong Han; Yang, Seo Yeon; You, Jihong; Lee, Sang Bae; You, Jin; Chang, Yoon Soo; Kim, Hyung Jung; Ahn, Chul Min; Byun, Min Kwang; Park, Jung-Won

    2016-01-01

    Background Specific immunoglobulin E (IgE) sensitization to staphylococcal enterotoxin (SE) has been recently considered to be related to allergic disease, including asthma. Despite studies on specific IgE (sIgE) to SE and its relationship to asthma diagnosis and severity, the association of sIgE to SE with airway hyperresponsiveness (AHR) remains unclear. Methods We enrolled 81 asthma patients admitted to the Severance Hospital in Korea from March 1, 2013, to February 28, 2015 and retrospectively reviewed the electronic medical records of the enrolled subjects. The serum levels of sIgE to SE (A/B) of all subjects was measured using the ImmunoCAP 250 (Phadia) system with SE-sIgE positive defined as >0.10 kU/mL. Results The SE-sIgE level was not significantly correlated with asthma severity (forced expiratory volume in 1 second [FEV1], FEV1/forced vital capacity, sputum eosinophils, and serum eosinophils), whereas the SE-sIgE level in patients with positive AHR (mean±standard error of the mean, 0.606±0.273 kU/mL) was significantly higher than that in patients with negative AHR (0.062±0.015 kU/mL, p=0.034). In regression analysis, SE sensitization (sIgE to SE ≥0.010 kU/mL) was a significant risk factor for AHR, after adjustment for age, sex, FEV1, and sputum eosinophils (odds ratio, 7.090; 95% confidence interval, 1.180–42.600; p=0.032). Prevalence of SE sensitization was higher in patients with allergic rhinitis and non-atopic asthma patients, as compared to patients without allergic rhinitis and atopic asthma patients, respectively, but without statistical significance. Conclusion SE sensitization is significantly associated with AHR.

  12. Linezolid-resistant staphylococcal bacteraemia: A multicentre case-case-control study in Italy.

    Science.gov (United States)

    Russo, Alessandro; Campanile, Floriana; Falcone, Marco; Tascini, Carlo; Bassetti, Matteo; Goldoni, Paola; Trancassini, Maria; Della Siega, Paola; Menichetti, Francesco; Stefani, Stefania; Venditti, Mario

    2015-03-01

    The aim of this multicentre study was to analyse the characteristics of patients with bloodstream infections due to staphylococcal strains resistant to linezolid. This was a retrospective case-case-control study of patients hospitalised in three large teaching hospitals in Italy. A linezolid-resistant (LIN-R) Staphylococcus spp. group and a linezolid-susceptible (LIN-S) Staphylococcus spp. group were compared with control patients to determine the clinical features and factors associated with isolation of LIN-R strains. All LIN-R Staphylococcus spp. strains underwent molecular typing. Compared with the LIN-S group, central venous catheters were the main source of infection in the LIN-R group. The LIN-R and LIN-S groups showed a similar incidence of severe sepsis or septic shock, and both showed a higher incidence of these compared with the control group. Overall, patients in the LIN-R group had a higher 30-day mortality rate. Multivariate analysis found previous linezolid therapy, linezolid therapy >14 days, antibiotic therapy in the previous 30 days, antibiotic therapy >14 days, previous use of at least two antibiotics and hospitalisation in the previous 90 days as independent risk factors associated with isolation of a LIN-R strain. The G2576T mutation in domain V of 23S rRNA was the principal mechanism of resistance; only one strain of Staphylococcus epidermidis carried the cfr methylase gene (A2503), together with L4 insertion (71GGR72) and L3 substitution (H146Q). LIN-R strains are associated with severe impairment of clinical conditions and unfavourable patient outcomes. Reinforcement of infection control measures may have an important role in preventing these infections.

  13. Pharmacokinetic/Toxicity Properties of the New Anti-Staphylococcal Lead Compound SK-03-92

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    William R. Schwan

    2015-11-01

    Full Text Available Because of the potential of a new anti-staphylococcal lead compound SK-03-92 as a topical antibiotic, a patch, or an orally active drug, we sought to determine its safety profile and oral bioavailability. SK-03-92 had a high IC50 (125 μg/mL in vitro against several mammalian cell lines, and mice injected intraperiteonally at the highest dose did not exhibit gross toxicity (e.g., altered gait, ungroomed, significant weight loss. Single dose (100 μg/g pharmacokinetic (PK analysis with formulated SK-03-92 showed that peak plasma concentration (1.64 μg/mL was achieved at 20–30 min. Oral relative bioavailability was 8%, and the drug half-life was 20–30 min, demonstrating that SK-03-92 is likely not a candidate for oral delivery. Five-day and two-week PK analyses demonstrated that SK-03-92 plasma levels were low. Multi-dose analysis showed no gross adverse effects to the mice and a SK-03-92 peak plasma concentration of 2.12 μg/mL with the presence of significant concentrations of breakdown products 15 min after dosing. SK-03-92 appeared to be very safe based on tissue culture and mouse gross toxicity determinations, but the peak plasma concentration suggests that a pro-drug of SK-03-92 or preparation of analogs of SK-03-92 with greater bioavailability and longer half-lives are warranted.

  14. Surface plasmon resonance detection of biological warfare agent Staphylococcal enterotoxin B using high affinity monoclonal antibody

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    Gupta, Garima; Singh, Pawan K.; Boopathi, M., E-mail: mannanboopathi@yahoo.com; Kamboj, D.V.; Singh, Beer; Vijayaraghavan, R.

    2010-11-30

    A novel sensitive method was developed for the detection as well as quantification of Staphylococcal enterotoxin B (SEB) using surface plasmon resonance (SPR). It is well known that the amount of SEB needed to cause the intoxication to human beings is very less and this concentration (0.02 {mu}g/kg) is highly dangerous, hence, it is used as biological warfare agent. Thus, the need to develop a reliable and potential detection system against SEB is warranted. In the present work, SEB antibody was immobilized on carboxymethyldextran modified gold chip. The immobilization of SEB antibody and interaction of antigen with immobilized antibody were in-situ characterized by SPR and electrochemical impedance spectroscopy. A sample solution containing SEB antigen was injected in a working channel and the results revealed linearity in the concentration from 2.0 to 32.0 pM with a detection limit of 1.0 pM. By using kinetic evaluation software, K{sub D} (equilibrium constant) and Bmax (maximum binding capacity of analyte) values were calculated and found to be 13 pM and 424.23, respectively. Moreover, the thermodynamic parameter, change in Gibb's free energy was deduced and found to be -62.08 kJ/mol and this value shows the spontaneous interaction between SEB antigen and SEB antibody. In order to optimize the detection method, temperature and pH variation studies were also performed. Interference study was conducted to know the selectivity for the antigen-antibody interaction of SEB. The selectivity efficiency of SEB, SEC, SEA and SED were 100, 27.15, 20.01 and 12.05%, respectively towards SEB antibody.

  15. Mechanisms mediating enhanced neutralization efficacy of staphylococcal enterotoxin B by combinations of monoclonal antibodies.

    Science.gov (United States)

    Dutta, Kaushik; Varshney, Avanish K; Franklin, Matthew C; Goger, Michael; Wang, Xiaobo; Fries, Bettina C

    2015-03-13

    Staphylococcal enterotoxin B (SEB) is a superantigen that cross-links the major histocompatibility complex class II and specific V-β chains of the T-cell receptor, thus forming a ternary complex. Developing neutralizing mAb to disrupt the ternary complex and abrogate the resulting toxicity is a major therapeutic challenge because SEB is effective at very low concentrations. We show that combining two SEB-specific mAbs enhances their efficacy, even though one of the two mAbs by itself has no effect on neutralization. Crystallography was employed for fine-mapping conformational epitopes in binary and ternary complexes between SEB and Fab fragments. NMR spectroscopy was used to validate and identify subtle allosteric changes induced by mAbs binding to SEB. The mapping of epitopes established that a combination of different mAbs can enhance efficacy of mAb-mediated protection from SEB induced lethal shock by two different mechanisms: one mAb mixture promoted clearance of the toxin both in vitro and in vivo by FcR-mediated cross-linking and clearance, whereas the other mAb mixture induced subtle allosteric conformational changes in SEB that perturbed formation of the SEB·T-cell receptor·major histocompatibility complex class II trimer. Finally structural information accurately predicted mAb binding to other superantigens that share conformational epitopes with SEB. Fine mapping of conformational epitopes is a powerful tool to establish the mechanism and optimize the action of synergistic mAb combinations.

  16. Tagging staphylococcal enterotoxin B (SEB) with TGFaL3 for breast cancer therapy.

    Science.gov (United States)

    Yousefi, Forough; Siadat, Seyed Davar; Saraji, Alireza Azizi; Hesaraki, Saeed; Aslani, Mohammad Mehdi; Mousavi, Seyed Fazlollah; Imani Fooladi, Abbas Ali

    2016-04-01

    Recent research has attempted to direct superantigens towards tumors by means of tumor-targeted superantigen (TTS) strategy. In this study, we explored the antitumor property of TTS by fusing the third loop of transforming growth factor α (TGFαL3) to staphylococcal enterotoxin type B (SEB) and investigated the possibility of the therapeutic application of TGFαL3-SEB as a novel antitumor candidate in mice bearing breast cancer. Treatment was performed through intratumoral and intravenous injection of TGFαL3-SEB. Tumor size/volume, long-term survival, and cytokine secretion were assessed. In addition, the toxicity of each treatment on liver and kidneys was examined. Our results indicated that the relative tumor volume significantly increased in the mice receiving intratumoral TGFaL3-SEB (p < 0.05). Surprisingly, 5 out of the 14 mice were cleared from the tumor thoroughly in 10-25 days after intratumoral administration of TGFaL3-SEB. Quantification of cytokines clearly showed that the mice receiving intratumoral SEB significantly secreted higher interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) compared with the other groups (p < 0.05). The antitumor effect was followed by inhibition of cell proliferation (Ki-67) and micro vascularization (CD31). The highest and lowest levels of tumor necrosis were observed in the intratumoral administration of TGFαL3-SEB (85 %) and PBS (14 %), respectively. Intratumoral injection of TGFαL3-SEB increased the lifespan of the mice so 37.5 % of them could survive for more than 6 months (p < 0.05). Overall, our findings indicated that intratumoral administration of TGFαL3-SEB effectively inhibited the growth of breast tumors through induction of necrosis and suppressing proliferation and angiogenesis without systemic toxicity.

  17. Staphylococcal enterotoxin B/texosomes as a candidate for breast cancer immunotherapy.

    Science.gov (United States)

    Imani Fooladi, Abbas Ali; Halabian, Raheleh; Mahdavi, Mehdi; Amin, Mohsen; Mahmoodzadeh Hosseini, Hamideh

    2016-01-01

    A new recombinant construct made up of two components, texosomes (TEX) and staphylococcal enterotoxin B (SEB), showed cytostatic properties against several types of tumor cells in vitro. Here, we aimed to assess the construct's antitumor immunogenicity in a murine tumor model. SEB was anchored onto purified texosomes and was used for immunization of mice before challenge with 4T1 cells. Tumor size, survival time, necrosis, metastasis rate, and the levels of IL-2, IL-4, IL-17, IL-12, TNF-α, and IFN-γ were measured. Immunization of the mice with TEX-SEB increased the stimulation index of splenocytes significantly compared with the PBS-treated mice (p < 0.01). In addition, there was a significant increase of TNF-α, IL-2, and IFN-γ secreted from isolated splenocytes of the mice immunized by either TEX-SEB, TEX + SEB, TEX, or SEB in comparison with PBS (p < 0.001, p < 0.001, and p < 0.05, respectively), whereas no significant change of IL-4 secretion was observed in any treated groups. Finding from tumor tissue homogenate testing showed that the level of IL-17 and IFN-γ among mice immunized with TEX-SEB was significantly lower than PBS-treated group (p < 0.05). IL-12, IL-4, and TNF-α levels were not significantly different from PBS- and TEX-SEB-immunized groups except in the SEB-immunized mice. Although TEX-SEB immunization relatively prolonged the survival of the mice, it had no inhibitory impact on tumor size. Pathologic manifestations showed the significant rise of necrosis after immunization with TEX-SEB compared to PBS (p < 0.01). Overall, our findings suggest that the presence of SEB rescues tumorigenesis effects of TEX making the construct an appropriate candidate for tumor immunotherapy.

  18. Rhinosinusitis derived Staphylococcal enterotoxin B possibly associates with pathogenesis of ulcerative colitis

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    An Zi-Yuan

    2005-09-01

    Full Text Available Abstract Background During clinical practice, we noticed that some patients with both ulcerative colitis (UC and chronic rhinosinusitis (CRS showed amelioration of UC after treatment of CRS. This study was designed to identify a possible association between CRS and UC. Methods Thirty-two patients with both CRS and UC received treatment with functional endoscopic sinus surgery (FESS for CRS. Clinical symptom scores for CRS and UC, as well as serum levels of anti-Staphylococcal enterotoxin B (SEB were evaluated at week 0 and week 12. Sinus wash fluid SEB content was measured with enzyme-linked immunosorbent assay (ELISA. The surgically removed tissues were cultured to identify growth of Staphylococcus. aureus (S. aureus. Immunohistochemistry was employed to identify anti-SEB positive cells in the colonic mucosa. Colonic biopsies were obtained and incubated with SEB. Mast cell activation in the colonic mucosa in response to incubation with SEB was observed with electron microscopy and immunoassay. Results The clinical symptom scores of CRS and UC severe scores (UCSS were significantly reduced in the UC-CRS patients after FESS. The number of cultured S. aureus colonies from the surgically removed sinus mucosa significantly correlated with the decrease in UCSS. High levels of SEB were detected in the sinus wash fluids of the patients with UC-CRS. Histamine and tryptase release was significantly higher in the culture supernate in the patients with UC-CRS than the patients with UC-only and normal controls. Anti-SEB positive cells were located in the colonic mucosa. Conclusion The pathogenesis of UC in some patients may be associated with their pre-existing CRS by a mechanism of swallowing sinusitis-derived SEB. We speculate that SEB initiates inappropriate immune reactions and inflammation in the colonic mucosa that further progresses to UC.

  19. MALDI-TOF-MS for rapid detection of staphylococcal Panton-Valentine leukocidin.

    Science.gov (United States)

    Bittar, Fadi; Ouchenane, Zoulikha; Smati, Farida; Raoult, Didier; Rolain, Jean-Marc

    2009-11-01

    Toxin-producing gram-positive bacteria are responsible for emerging and life-threatening infections in humans worldwide. Both rapid toxin detection and adapted therapy are essential to limit the morbidity due to such toxins, especially staphylococcal Panton-Valentine leukocidin (PVL). Here we describe the use of a mass spectrometry profile generated by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) followed by ClinProTools 2.0 software analysis to find a reproducible model able to identify PVL in Staphylococcus aureus strains. Eighty-one S. aureus strains were used and tested for the presence of PVL, toxic shock syndrome toxin (TSST-1) and mecA genes. The peak at 4448 mass-to-charge ratio (m/z) was the most relevant peak to differentiate between PVL-producing and non-PVL-producing S. aureus. A model using only this peak had an overall recognition capability of 100% and an overall cross-validation of 77.07%. Prospective evaluation of the model allowed two cases of PVL-producing strains to be detected within a few minutes during the time of care and before polymerase chain reaction (PCR) results. Our study represents a proof of concept for the use of such rapid technology as a point-of-care method to identify potential lethal toxin quickly. We believe that such a rapid method will be timely to help change the therapeutic strategy and could be used in the future for other pathogens and infectious diseases.

  20. Modeling the effects of a Staphylococcal Enterotoxin B (SEB on the apoptosis pathway

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    Hammamieh Rasha

    2006-05-01

    Full Text Available Abstract Background The lack of detailed understanding of the mechanism of action of many biowarfare agents poses an immediate challenge to biodefense efforts. Many potential bioweapons have been shown to affect the cellular pathways controlling apoptosis 1234. For example, pathogen-produced exotoxins such as Staphylococcal Enterotoxin B (SEB and Anthrax Lethal Factor (LF have been shown to disrupt the Fas-mediated apoptotic pathway 24. To evaluate how these agents affect these pathways it is first necessary to understand the dynamics of a normally functioning apoptosis network. This can then serve as a baseline against which a pathogen perturbed system can be compared. Such comparisons can expose both the proteins most susceptible to alteration by the agent as well as the most critical reaction rates to better instill control on a biological network. Results We explore this through the modeling and simulation of the Fas-mediated apoptotic pathway under normal and SEB influenced conditions. We stimulated human Jurkat cells with an anti-Fas antibody in the presence and absence of SEB and determined the relative levels of seven proteins involved in the core pathway at five time points following exposure. These levels were used to impute relative rate constants and build a quantitative model consisting of a series of ordinary differential equations (ODEs that simulate the network under both normal and pathogen-influenced conditions. Experimental results show that cells exposed to SEB exhibit an increase in the rate of executioner caspase expression (and subsequently apoptosis of 1 hour 43 minutes (± 14 minutes, as compared to cells undergoing normal cell death. Conclusion Our model accurately reflects these results and reveals intervention points that can be altered to restore SEB-influenced system dynamics back to levels within the range of normal conditions.

  1. Construction and characterization of VL-VH tail-parallel genetically engineered antibodies against staphylococcal enterotoxins.

    Science.gov (United States)

    He, Xianzhi; Zhang, Lei; Liu, Pengchong; Liu, Li; Deng, Hui; Huang, Jinhai

    2015-03-01

    Staphylococcal enterotoxins (SEs) produced by Staphylococcus aureus have increasingly given rise to human health and food safety. Genetically engineered small molecular antibody is a useful tool in immuno-detection and treatment for clinical illness caused by SEs. In this study, we constructed the V(L)-V(H) tail-parallel genetically engineered antibody against SEs by using the repertoire of rearranged germ-line immunoglobulin variable region genes. Total RNA were extracted from six hybridoma cell lines that stably express anti-SEs antibodies. The variable region genes of light chain (V(L)) and heavy chain (V(H)) were cloned by reverse transcription PCR, and their classical murine antibody structure and functional V(D)J gene rearrangement were analyzed. To construct the eukaryotic V(H)-V(L) tail-parallel co-expression vectors based on the "5'-V(H)-ivs-IRES-V(L)-3'" mode, the ivs-IRES fragment and V(L) genes were spliced by two-step overlap extension PCR, and then, the recombined gene fragment and V(H) genes were inserted into the pcDNA3.1(+) expression vector sequentially. And then the constructed eukaryotic expression clones termed as p2C2HILO and p5C12HILO were transfected into baby hamster kidney 21 cell line, respectively. Two clonal cell lines stably expressing V(L)-V(H) tail-parallel antibodies against SEs were obtained, and the antibodies that expressed intracytoplasma were evaluated by enzyme-linked immunosorbent assay, immunofluorescence assay, and flow cytometry. SEs can stimulate the expression of some chemokines and chemokine receptors in porcine IPEC-J2 cells; mRNA transcription level of four chemokines and chemokine receptors can be blocked by the recombinant SE antibody prepared in this study. Our results showed that it is possible to get functional V(L)-V(H) tail-parallel genetically engineered antibodies in same vector using eukaryotic expression system.

  2. The interactions of porcine and ovine, serum and colostral immunoglobulins with staphylococcal Protein A.

    Science.gov (United States)

    Bennell, M A; Watson, D L

    1980-01-01

    The purpose of these studies was to determine the proportion of each immunoglobulin class/subclass in blood and colostrum of the pig and sheep, which would bind to staphylococcal Protein A. The concentrations of porcine IgG, IgM, and IgA were determined for serum and colostral whey from five sows. Similar measurements were made on two fractions produce by elution of the sample through a Protein A-Sepharose column: fraction 1, immunoglobulins which did not bind to Protein A, and fraction 2, immunoglobulins which bound to Protein A. The concentrations of ovine IgG1, IgG2, IgM, and IgA were measured for serum and colostral whey from six ewes, and again similar measurements were made after elution of each ovine sample through Protein A-Sepharose. All classes/subclasses of porcine and ovine serum and colostral immunoglobulins bound to Protein A to some extent. More than 90% of IgG from both porcine colostral whey and serum bound to Protein A. Ovine IgG1 from most ewes possessed a low affinity for Protein A whereas ovine IgG2 generally possessed a high affinity; 100% of the IgG2 in ovine colostral whey samples bound to Protein A. There was remarkable variation between individuals in the binding capacity of porcine IgM and each of the ovine immunoglobulins. For the ovine samples, in particular there were distinct differences between Protein A binding capacity of serum and colostral immunoglobulins of the same class/subclass.

  3. Characterization of fatty acid modifying enzyme activity in staphylococcal mastitis isolates and other bacteria

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    Lu Thea

    2012-06-01

    Full Text Available Abstract Background Fatty acid modifying enzyme (FAME has been shown to modify free fatty acids to alleviate their bactericidal effect by esterifying fatty acids to cholesterol or alcohols. Although it has been shown in previous studies that FAME is required for Staphylococcus aureus survival in skin abscesses, FAME is poorly studied compared to other virulence factors. FAME activity had also been detected in coagulase-negative staphylococci (CNS. However, FAME activity was only surveyed after a bacterial culture was grown for 24 h. Therefore if FAME activity was earlier in the growth phase, it would not have been detected by the assay and those strains would have been labeled as FAME negative. Results Fifty CNS bovine mastitis isolates and several S. aureus, Escherichia coli, and Streptococcus uberis strains were assayed for FAME activity over 24 h. FAME activity was detected in 54% of CNS and 80% S. aureus strains surveyed but none in E. coli or S. uberis. While some CNS strains produced FAME activity comparable to the lab strain of S. aureus, the pattern of FAME activity varied among strains and across species of staphylococci. All CNS that produced FAME activity also exhibited lipase activity. Lipase activity relative to colony forming units of these CNS decreased over the 24 h growth period. No relationship was observed between somatic cell count in the milk and FAME activity in CNS. Conclusions Some staphylococcal species surveyed produced FAME activity, but E. coli and S. uberis strains did not. All FAME producing CNS exhibited lipase activity which may indicate that both these enzymes work in concert to alter fatty acids in the bacterial environment.

  4. Staphylococcal protein Ecb impairs complement receptor-1 mediated recognition of opsonized bacteria.

    Science.gov (United States)

    Amdahl, Hanne; Haapasalo, Karita; Tan, Lydia; Meri, Taru; Kuusela, Pentti I; van Strijp, Jos A; Rooijakkers, Suzan; Jokiranta, T Sakari

    2017-01-01

    Staphyloccus aureus is a major human pathogen leading frequently to sepsis and soft tissue infections with abscesses. Multiple virulence factors including several immune modulating molecules contribute to its survival in the host. When S. aureus invades the human body, one of the first line defenses is the complement system, which opsonizes the bacteria with C3b and attract neutrophils by release of chemotactic peptides. Neutrophils express Complement receptor-1 [CR1, CD35) that interacts with the C3b-opsonized particles and thereby plays an important role in pathogen recognition by phagocytic cells. In this study we observed that a fraction of S. aureus culture supernatant prevented binding of C3b to neutrophils. This fraction consisted of S. aureus leukocidins and Efb. The C-terminus of Efb is known to bind C3b and shares significant sequence homology to the extracellular complement binding protein [Ecb). Here we show that S. aureus Ecb displays various mechanisms to block bacterial recognition by neutrophils. The presence of Ecb blocked direct interaction between soluble CR1 and C3b and reduced the cofactor activity of CR1 in proteolytic inactivation of C3b. Furthermore, Ecb could dose-dependently prevent recognition of C3b by cell-bound CR1 that lead to impaired phagocytosis of NHS-opsonized S. aureus. Phagocytosis was furthermore reduced in the presence of soluble CR1 [sCR1). These data indicate that the staphylococcal protein Ecb prevents recognition of C3b opsonized bacteria by neutrophil CR1 leading to impaired killing by phagocytosis and thereby contribute to immune evasion of S. aureus.

  5. Improved purification and biologic activities of staphylococcal toxic shock syndrome toxin 1.

    Science.gov (United States)

    Kum, W W; Laupland, K B; See, R H; Chow, A W

    1993-01-01

    An improved method for producing highly purified toxic shock syndrome toxin 1 (TSST-1) by preparative isoelectric focusing in a Bio-Rad Rotofor cell and then chromatofocusing is described. Purification to homogeneity was confirmed by silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE; 50 micrograms of protein was loaded), by immunoblotting with polyclonal rabbit antiserum raised against the crude culture supernatant used for purification, and by autoradiography after iodination and SDS-PAGE. Biologic activity was demonstrated by mitogenicity and cytokine induction (tumor necrosis factor alpha [TNF-alpha], interleukin 1-beta [IL-1 beta], and IL-6) of human peripheral blood mononuclear cells (PBMCs) and by lethality in New Zealand White rabbits following subcutaneous infusion. In contrast to commercial TSST-1 preparations, our TSST-1 preparation required the presence of both monocytes and T cells for the induction of TNF-alpha and IL-1 beta from human PBMCs. A 46-kDa contaminating protein in the commercial TSST-1 preparation, identified as staphylococcal lipase, was likely responsible for the induction of TNF-alpha and IL-1 beta from human monocytes in the absence of T cells, a biologic activity falsely attributed to purified TSST-1. Our improved purification procedure for TSST-1 provides a high yield and is both more rapid and less labor intensive than previously reported methods. Furthermore, our studies clearly demonstrate the need for stringent methods of purity assessment of TSST-1 preparations before ascribing to them their potent biologic activities. Images PMID:8253961

  6. Regulation of staphylococcal enterotoxin B-elicited nitric oxide production by endothelial cells.

    Science.gov (United States)

    LeClaire, R D; Kell, W M; Sadik, R A; Downs, M B; Parker, G W

    1995-01-01

    The effect of staphylococcal enterotoxin B (SEB)-elicited inducible nitric oxide synthase (iNOS) in mouse endothelial cells was investigated. Results showed that SEB stimulated the same level of NO production in gamma interferon (IFN-gamma)-primed cells as did trichloroacetic acid-extracted lipopolysaccharide. The kinetics of induced NO production and expression of mRNA for iNOS differed markedly in endothelial and macrophage cells. Induced endothelial nitrite production was transient and was 15 to 20% of that generated by macrophage cells; mRNA levels peaked by 2 h and then steadily declined, whereas macrophage message levels continually increased. The ability of endothelial cells to produce SEB-induced NO depended on priming with IFN-gamma, although detectable mRNA could be elicited by SEB alone. Induction of endothelial iNOS mRNA was inhibited by cycloheximide, which indicated a requirement for de novo protein synthesis. Niacinamide and interleukin-10 significantly reduced SEB-induced endothelial NO production. Both are reported to affect IFN-gamma-induced class II major histocompatibility complex (MHC) expression on antigen-presenting cells. Niacinamide reduced iNOS mRNA levels and markedly reduced IFN-gamma induction of endothelial class II MHC surface antigen. Interleukin-10 did not consistently reduce iNOS mRNA expression and had no effect on IFN-gamma induction of endothelial class II MHC surface antigen. These results suggest that SEB interacts with IFN-gamma-primed endothelial cells to elicit induced NO and that this induction can be effectively modulated at the receptor or transcriptional level. PMID:7529748

  7. Staphylococcal Bicomponent Pore-Forming Toxins: Targets for Prophylaxis and Immunotherapy

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    M. Javad Aman

    2014-03-01

    Full Text Available Staphylococccus aureus represents one of the most challenging human pathogens as well as a common colonizer of human skin and mucosal surfaces. S. aureus causes a wide range of diseases from skin and soft tissue infection (SSTI to debilitating and life-threatening conditions such as osteomyelitis, endocarditis, and necrotizing pneumonia. The range of diseases reflects the remarkable diversity of the virulence factors produced by this pathogen, including surface antigens involved in the establishment of infection and a large number of toxins that mediate a vast array of cellular responses. The staphylococcal toxins are generally believed to have evolved to disarm the innate immune system, the first line of defense against this pathogen. This review focuses on recent advances on elucidating the biological functions of S. aureus bicomponent pore-forming toxins (BCPFTs and their utility as targets for preventive and therapeutic intervention. These toxins are cytolytic to a variety of immune cells, primarily neutrophils, as well as cells with a critical barrier function. The lytic activity of BCPFTs towards immune cells implies a critical role in immune evasion, and a number of epidemiological studies and animal experiments relate these toxins to clinical disease, particularly SSTI and necrotizing pneumonia. Antibody-mediated neutralization of this lytic activity may provide a strategy for development of toxoid-based vaccines or immunotherapeutics for prevention or mitigation of clinical diseases. However, certain BCPFTs have been proposed to act as danger signals that may alert the immune system through an inflammatory response. The utility of a neutralizing vaccination strategy must be weighed against such immune-activating potential.

  8. Staphylococcal bicomponent pore-forming toxins: targets for prophylaxis and immunotherapy.

    Science.gov (United States)

    Aman, M Javad; Adhikari, Rajan P

    2014-03-04

    Staphylococccus aureus represents one of the most challenging human pathogens as well as a common colonizer of human skin and mucosal surfaces. S. aureus causes a wide range of diseases from skin and soft tissue infection (SSTI) to debilitating and life-threatening conditions such as osteomyelitis, endocarditis, and necrotizing pneumonia. The range of diseases reflects the remarkable diversity of the virulence factors produced by this pathogen, including surface antigens involved in the establishment of infection and a large number of toxins that mediate a vast array of cellular responses. The staphylococcal toxins are generally believed to have evolved to disarm the innate immune system, the first line of defense against this pathogen. This review focuses on recent advances on elucidating the biological functions of S. aureus bicomponent pore-forming toxins (BCPFTs) and their utility as targets for preventive and therapeutic intervention. These toxins are cytolytic to a variety of immune cells, primarily neutrophils, as well as cells with a critical barrier function. The lytic activity of BCPFTs towards immune cells implies a critical role in immune evasion, and a number of epidemiological studies and animal experiments relate these toxins to clinical disease, particularly SSTI and necrotizing pneumonia. Antibody-mediated neutralization of this lytic activity may provide a strategy for development of toxoid-based vaccines or immunotherapeutics for prevention or mitigation of clinical diseases. However, certain BCPFTs have been proposed to act as danger signals that may alert the immune system through an inflammatory response. The utility of a neutralizing vaccination strategy must be weighed against such immune-activating potential.

  9. Is Staphylococcal Screening and Suppression an Effective Interventional Strategy for Reduction of Surgical Site Infection?

    Science.gov (United States)

    Edmiston, Charles E; Ledeboer, Nathan A; Buchan, Blake W; Spencer, Maureen; Seabrook, Gary R; Leaper, David

    2016-04-01

    Staphylococcus aureus has been recognized as a major microbial pathogen for over 100 y, having the capacity to produce a variety of suppurative and toxigenic disease processes. Many of these infections are life-threatening, with particularly enhanced virulence in hospitalized patients with selective risk factors. Strains of methicillin-resistant Staphylococcus aureus (MRSA) have rapidly spread throughout the healthcare environment such that approximately 20% of S. aureus isolates recovered from surgical site infections are methicillin-resistant, (although this is now reducing following national screening and suppression programs and high impact interventions). Widespread nasal screening to identify MRSA colonization in surgical patients prior to admission are controversial, but selective, evidence-based studies have documented a reduction of surgical site infection (SSI) after screening and suppression. Culture methods used to identify MRSA colonization involve selective, differential, or chromogenic media. These methods are the least expensive, but turnaround time is 24-48 h. Although real-time polymerase chain reaction (RT-PCR) technology provides rapid turnaround (1-2 h) with exceptional testing accuracy, the costs can range from three to 10 times more than conventional culture methodology. Topical mupirocin, with or without pre-operative chlorhexidine showers or skin wipes, is the current "gold-standard" for nasal decolonization, but inappropriate use of mupirocin is associated with increasing staphylococcal resistance. Selection of an effective active universal or targeted surveillance strategy should be based upon the relative risk of MSSA or MRSA surgical site infection in patients undergoing orthopedic or cardiothoracic device related surgical procedures.

  10. MCCE analysis of the pKas of introduced buried acids and bases in staphylococcal nuclease.

    Science.gov (United States)

    Gunner, M R; Zhu, Xuyu; Klein, Max C

    2011-12-01

    The pK(a)s of 96 acids and bases introduced into buried sites in the staphylococcal nuclease protein (SNase) were calculated using the multiconformation continuum electrostatics (MCCE) program and the results compared with experimental values. The pK(a)s are obtained by Monte Carlo sampling of coupled side chain protonation and position as a function of pH. The dependence of the results on the protein dielectric constant (ε(prot)) in the continuum electrostatics analysis and on the Lennard-Jones non-electrostatics parameters was evaluated. The pK(a)s of the introduced residues have a clear dependence on ε(prot,) whereas native ionizable residues do not. The native residues have electrostatic interactions with other residues in the protein favoring ionization, which are larger than the desolvation penalty favoring the neutral state. Increasing ε(prot) scales both terms, which for these residues leads to small changes in pK(a). The introduced residues have a larger desolvation penalty and negligible interactions with residues in the protein. For these residues, changing ε(prot) has a large influence on the calculated pK(a). An ε(prot) of 8-10 and a Lennard-Jones scaling of 0.25 is best here. The X-ray crystal structures of the mutated proteins are found to provide somewhat better results than calculations carried out on mutations made in silico. Initial relaxation of the in silico mutations by Gromacs and extensive side chain rotamer sampling within MCCE can significantly improve the match with experiment. Copyright © 2011 Wiley-Liss, Inc.

  11. Staphylococcal Panton-Valentine leucocidin as a major virulence factor associated to furuncles.

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    Lamine Baba-Moussa

    Full Text Available Panton-Valentine Leucocidin (PVL, one of the β-barrel pore-forming staphylococcal leucotoxins, is known to be associated to furuncles and some severe community pneumonia. However, it is still uncertain how many other virulence factors are also associated to furuncles and what the risk factors of furuncles are in immuno-compromised status of patients, especially the HIV (+ patients. In this paper, we use antigen immunoprecipitation and multiplex PCR approach to determine the presence of 19 toxins, 8 adhesion factors and the PFGE profiles associated to furuncles in three independent patient study groups of S. aureus (SA isolates collected from the Cayenne General Hospital (French Guiana. The patient groups were made of: 16 isolates from HIV (- patients, 9 from HIV (+ patients suffering from furuncles, and 30 control isolates from patients with diverse secondary infected dermatitis. Our data reveals that the majority (96% of SA strains isolated from HIV patient-derived furuncles significantly produced PVL (p<10(-7, whereas only 10% of SA strains produced this toxin in secondary infected dermatosis. A high prevalence of LukE-LukD-producing isolates (56 to 78% was recorded in patient groups. Genes encoding clumping factor B, collagen- and laminin-binding proteins (clfB, cna, lbp, respectively were markedly frequent (30 to 55%, without being associated to a specific group. Pulse field gel electrophoresis evidenced 24 overall pulsotypes, whereas the 25 PVL-producing isolates were distributed into 15 non clonal fingerprints. These pulsotypes were not specific PVL-producing isolates. PVL appears to be the major virulence factor associated to furuncles in Europe and in South America regardless of the immune status of the HIV patients.

  12. Uncoupling of T Cell Receptor Zeta Chain Function during the Induction of Anergy by the Superantigen, Staphylococcal Enterotoxin A

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    William D. Cornwell

    2010-06-01

    Full Text Available Staphylococcus aureus enterotoxins have immunomodulatory properties. In this study, we show that Staphylococcal enterotoxin A (SEA induces a strong proliferative response in a murine T cell clone independent of MHC class II bearing cells. SEA stimulation also induces a state of hypo-responsiveness (anergy. We characterized the components of the T cell receptor (TCR during induction of anergy by SEA. Most interestingly, TCR zeta chain phosphorylation was absent under SEA anergizing conditions, which suggests an uncoupling of zeta chain function. We characterize here a model system for studying anergy in the absence of confounding costimulatory signals.

  13. Use of a staphylococcal vaccine to reduce prevalence of mastitis and lower somatic cell counts in a registered Saanen dairy goat herd.

    Science.gov (United States)

    Kautz, F M; Nickerson, S C; Ely, L O

    2014-08-01

    This investigation evaluated the efficacy of a bacterin in reducing the prevalence of staphylococcal mastitis and somatic cell counts (SCC) in a dairy goat herd. Does were vaccinated or left as controls, and the levels of mastitis and SCC monitored over 18 months. Staphylococcus caprae (42.5%), S. xylosus (15.1%), and S. simulans (10.0%) were the predominant causes of intramammary infections (IMI). The infection rate was 1.64 IMI/doe among vaccinates, which tended to be lower (P mastitis vaccines for use in managing staphylococcal mastitis and SCC in dairy goats. Published by Elsevier Ltd.

  14. MEBT/MEBO治疗足部炽热沥青烫伤例临床观察%Clinical Observation of Treating 42 Cases of Broiling Bitumen Caused Foot Scald with MEBT/MEBO

    Institute of Scientific and Technical Information of China (English)

    钟海文; 董玉强; 王希龙; 罗若谷; 张延兵; 毛曾喜; 柳育才

    2013-01-01

      目的观察烧伤湿性医疗技术(MEBT/MEBO)治疗足部炽热沥青深度烫伤创面的临床效果。方法本组42例足部深度烫伤创面均采用MEBT/MEBO治疗,同时配合主、被动功能锻炼、按摩等综合康复措施治疗,连续临床观察半年评定疗效。结果患者创面全部一期愈合,经半年以上随访,愈合后的皮肤组织柔软,弹性较好,无瘢痕及功能障碍。结论MEBT/MEBO能始终保持创面湿润而不浸渍,坏死组织易被清除,疼痛减轻,愈合后的皮肤组织弹性较好,功能恢复理想。%Objective To observe the clinical therapeutic effects of MEBT/MEBO in treating foot deep scald caused by broiling bitumen. Methods A total of 42 cases with foot deep scald were treated with MEBT/MEBO, and at the meanwhile, some comprehensive rehabilitating treatments such as active and passive functional exercises and massage were performed conjunctively. Clinical observation was carried out continuously for half a year to evaluate the therapeutic effects. Results All the wounds got primary healing. Based on the follow-up of more than half a year, the healed skin tissue was soft with good elasticity, no scar and dysfunction. Conclusion MEBT/MEBO can always maintain the wound moist without macerating it, remove necrotic tissue easily and reduce pain. The healed skin tissue has better elasticity and ideal recovered function.

  15. 丹曲林对严重烧伤大鼠骨骼肌损害的治疗作用及机制%Effect and mechanism of dantrolene on skeletal muscle of rats with severe scald injury

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Objective To explore the effect and mechanism of ryanodine receptor antagonist dantrolene on skeletal muscle of rats with severe scald injury.Methods A total of 56 Wistar rats were divided into control , scald and dantrolene treatment groups according to a random digital table.Rats in scald and dantrolene treatment groups were subject to 50%total body surface area ( TBSA) full-thickness scald by a 12-second immersion of back and a 6-second immersion of abdomen in 94 ℃water and then received an intraperitoneal injection of Ringer′s solution.At the same time , the rats in scald group received 5%mannitol through caudal vein while those in dantrolene treatment group received dantrolene 2 mg/kg ( dissolved in 5%mannitol ).Rats in control group were sham-injured through an immersion of back and abdomen into 37 ℃warm water.Tibialis anterior muscle samples were harvested at Days 1, 4 and 7 post-scalding.Changes of skeletal muscle ultrastructure were observed by transmission electron microscope , subcellular calcium ion ( Ca2+) contents of skeletal muscle ( including cytoplasm , mitochondria & sarcoplasm reticulum ) were detected by electron probe X-ray microanalysis ( EPMA) and the levels of calpain-1 and calpain-2 protein were determined by Western blot.And the activities of calpain were detected by enzyme-linked immunosorbent assay.Results In scald group , assorted arrangement appeared immediately at Day 1 post-injury and partial disappearance of Z lines at Day 7 post-injury.There were no significant ultrastructure changes in dantrolene treatment group at Day 1 and 4 post-injury.Curled filament and mild fracture occurred merely in dantrolene treatment group at Day 7 post-injury.The cytoplasmic contents of Ca 2+ were significantly higher in scald group than those in control group at Day 1 and 4 ((0.964 ±0.060), (0.639 ± 0.067) vs (0.266 ±0.029) μmol/L respectively, all P0.05).Caplain-1 and calpain-2 protein levels in scald group increased significantly at Day 1

  16. Magnetic Bead and Fluorescent Silica Nanoparticles Based Optical Immunodetection of Staphylococcal Enterotoxin B (SEB in Bottled Water

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    Shiva K. RASTOGI

    2009-10-01

    Full Text Available Staphylococcal enterotoxins (SEs are a major cause of food-borne diseases, most commonly SEs assayed immunologically with ELISA. An immunoassay based on fluorescein dye doped silica dioxide nanoparticles (F-SiNPs and magnetic bead (MB is described here for the detection of staphylococcal enterotoxin B (SEB. F-SiNPs have unique optical properties which make them attractive for biosensing. The water-in-oil (W/O reverse microemulsion method was used for the synthesis of F-SiNPs (~ 95 nm of diameter. The F-SiNPs were characterized using SEM, TEM and FTIR spectroscopy. The detection of SEB is preformed in PBS buffer, and bottled drinking water using sandwich immunoassay format. Target analytes were captured using MBs modified with the antigen-specific “capture” antibody, and detected using F-SiNP labeled secondary antigen-specific antibody. We report a limit of detection down to 1 ng/mL SEB spiked sample in less than 2 hr assay time using fluorocount method. This study demonstrates the bio warfare agent SEB capture by magnetic beads and detection using F-SiNPs.

  17. Determination of anti-staphylococcal activity of thymoquinone in combinations with antibiotics by checkerboard method using EVA capmat™ as a

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    Johana Rondevaldova

    2017-05-01

    Full Text Available Thymoquinone (Tq has been reported to potentiate the in vitro growth-inhibitory activity of some antibiotics especially against Staphylococcus aureus. However, it has been shown that Tq vapors can affect the results of susceptibility testing by standard broth microdilution method. Therefore, we made a comparative experiment with and without ethylene vinyl acetate cap mats (EVA capmat™ on microplates. The results showed significant differences in the minimum inhibitory concentration values and proved this capmat as an effective vapor barrier. Therefore further experiments focused on the in vitro anti-staphylococcal combinatory effect of Tq with oxacillin, penicillin, and tetracycline against various S. aureus strains have been performed by checkerboard method using EVA capmat™. The combined effect was evaluated according to the sum of fractional inhibitory concentrations (ΣFIC. Synergy was obtained for combination with oxacillin against 3 (ΣFIC 0.263–0.450, with penicillin against 1 (ΣFIC 0.466 and with tetracycline against 2 strains tested (ΣFIC 0.400–0.475. Our results confirm previous reports on the Tq enhancement of anti-staphylococcal activity of antibiotics. Moreover, this is the first report on Tq synergy with oxacillin and penicillin against S. aureus. Our experiments also showed that Tq vapors can affect evaluation of combined effect by checkerboard assay, whereas the use of EVA capmat™ can avoid this.

  18. Staphylococcal enterotoxin genes are common in Staphylococcus aureus intestinal flora in Sudden Infant Death Syndrome (SIDS) and live comparison infants.

    Science.gov (United States)

    Highet, Amanda R; Goldwater, Paul N

    2009-11-01

    Pathological and epidemiological findings in sudden infant death syndrome (SIDS) suggest an infectious aetiology with indications of involvement of staphylococcal enterotoxins (SEs). While SEA, SEB and SEC have been found in the sera and tissues of SIDS cases, little is known about the role of intestinal Staphylococcus aureus or the roles of later-described toxins SEE, SEG, SEH, SEI and SEJ in SIDS. We used a molecular-based approach to define whether the intestinal tract could be a source of SEs to support the staphylococcal toxic shock hypothesis for SIDS. Intestinal contents from 57 SIDS infants and faeces from 79 age- and gender-matched live comparison infants were cultured and tested for S. aureus and sea-b-c-e-g-h-j and TSST using PCR. High proportions of infants in both groups carried toxigenic and nontoxigenic S. aureus. Significantly greater proportions of SIDS compared with comparison babies were positive for S. aureus (68.4% vs. 40.5%) and for SE genes (43.8% vs. 21.5%), suggesting a possible role in SIDS. The results indicate that colonization by S. aureus with SE genes is common in infants; however, their detection is unlikely to be a strong predictive tool for SIDS. Other factors (including immune response) may reveal a specific susceptibility to SEs in SIDS infants.

  19. Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Ito, T; Katayama, Y; Asada, K; Mori, N; Tsutsumimoto, K; Tiensasitorn, C; Hiramatsu, K

    2001-05-01

    The beta-lactam resistance gene mecA of Staphylococcus aureus is carried by a novel mobile genetic element, designated staphylococcal cassette chromosome mec (SCCmec), identified in the chromosome of a Japanese methicillin-resistant S. aureus (MRSA) strain. We now report identification of two additional types of mecA-carrying genetic elements found in the MRSA strains isolated in other countries of the world. There were substantial differences in the size and nucleotide sequences between the elements and the SCCmec. However, new elements shared the chromosomal integration site with the SCCmec. Structural analysis of the new elements revealed that they possessed all of the salient features of the SCCmec: conserved terminal inverted repeats and direct repeats at the integration junction points, conserved genetic organization around the mecA gene, and the presence of cassette chromosome recombinase (ccr) genes responsible for the movements of SCCmec. The elements, therefore, were considered to comprise the SCCmec family of staphylococcal mobile genetic elements together with the previously identified SCCmec. Among 38 epidemic MRSA strains isolated in 20 countries, 34 were shown to possess one of the three typical SCCmec elements on the chromosome. Our findings indicated that there are at least three distinct MRSA clones in the world with different types of SCCmec in their chromosome.

  20. Recombinant Staphylococcal protein A with cysteine residue for preparation of affinity chromatography stationary phase and immunosensor applications

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    Gorbatiuk O. B.

    2015-04-01

    Full Text Available Aim. Engineering of recombinant Staphylococcal protein A with cysteine residue (SPA-Cys for preparation of affinity chromatography stationary phase and formation of bioselective element of immunosensor. Methods. DNA sequences encoding IgG-binding region of SPA, His-tag and cysteine were genetically fused and expressed in E. coli. SPA-Cys was immobilized on maleimide-functionalized silica beads for affinity chromatography stationary phase preparation and on a gold sensor surface as a bioselective element of immunosensor. Results. SPA-Cys was expressed at a high-level in a soluble form. The target protein was purified and showed a high IgG-binding activity. The capacity of the obtained SPA-Cys-based affinity chromatography stationary phase was 10–12 mg of IgG /ml. The purity of eluted IgG was more than 95 % in one-step purification procedure. The developed SPA-Cys-based bioselective element of immunosensor selectively interacted with human IgG and did not interact with the control proteins. Conclusions. The recombinant Staphylococcal protein A with cysteine residue was successfully used for the preparation of affinity chromatography stationary phase and formation of the bioselective element of immunosensor.

  1. Staphylococcal biofilm formation on the surface of three different calcium phosphate bone grafts: a qualitative and quantitative in vivo analysis.

    Science.gov (United States)

    Furustrand Tafin, Ulrika; Betrisey, Bertrand; Bohner, Marc; Ilchmann, Thomas; Trampuz, Andrej; Clauss, Martin

    2015-03-01

    Differences in physico-chemical characteristics of bone grafts to fill bone defects have been demonstrated to influence in vitro bacterial biofilm formation. Aim of the study was to investigate in vivo staphylococcal biofilm formation on different calcium phosphate bone substitutes. A foreign-body guinea-pig infection model was used. Teflon cages prefilled with β-tricalcium phosphate, calcium-deficient hydroxyapatite, or dicalcium phosphate (DCP) scaffold were implanted subcutaneously. Scaffolds were infected with 2 × 10(3) colony-forming unit of Staphylococcus aureus (two strains) or S. epidermidis and explanted after 3, 24 or 72 h of biofilm formation. Quantitative and qualitative biofilm analysis was performed by sonication followed by viable counts, and microcalorimetry, respectively. Independently of the material, S. aureus formed increasing amounts of biofilm on the surface of all scaffolds over time as determined by both methods. For S. epidermidis, the biofilm amount decreased over time, and no biofilm was detected by microcalorimetry on the DCP scaffolds after 72 h of infection. However, when using a higher S. epidermidis inoculum, increasing amounts of biofilm were formed on all scaffolds as determined by microcalorimetry. No significant variation in staphylococcal in vivo biofilm formation was observed between the different materials tested. This study highlights the importance of in vivo studies, in addition to in vitro studies, when investigating biofilm formation of bone grafts.

  2. Anti-Staphylococcal and wound healing activities of Ganoderma praelongum and Glycyrrhiza glabra formulation in mice

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    Abdulghani - Ameri

    2013-02-01

    Full Text Available Summary: Topical antimicrobial therapy is one of the most important methods of wound care. In this investigation, we evaluated topical gel preparations of Ganoderma praelongum and Glycyrrhiza glabra aqueous extracts alone and in combination for antimicrobial and wound healing activities in MRSA - infected excision and incision wound models in mice. Gel formulations containing 0.3% G. praelongum, 2.5% G. glabra and combination of the two were prepared and tested for their influence on wound infection, wound contraction and epithelization phases of wound healing. Mupirocin ointment was applied as a standard treatment antibiotic. Results indicated that all three gel formulations promoted wound healing in both models by influencing wound contraction and epithelization phases. Examination of wounds at the end of our study period revealed that wounds treated with G. praelongum and G. glabra gel  formulations (combination gel showed considerable contraction and epithelization as compared to the gel base – treated group (negative control. Assumption could be made that this wound promotion is due to the anti – Staphylococcal activity of Ganoderma and wound healing activity of G. glabra.   Industrial relevance: Medicine using fungal metabolites is now recognized. A mushroom that has gained worldwide attention is Ganoderma lucidum and has been used to treat a wide spectrum of ailments including fungal and bacterial infections for long periods of time throughout history of ancient traditional medicine. Previous reports indicate that ganoderma lucidum mycelia are usually prescribed in the form of soup, syrup, tea, tablets, capsules, tincture or bolus or injected as a solution of powdered spores. In this study, the combination gel containing aqueous extracts of G. praelongum and G. glabra effectively inhibited the growth of methicillin resistant Staphylococcus aureus and promoted wound healing in mice. Further phytochemical studies are needed to

  3. Staphylococcal enterotoxin A regulates bone marrow granulocyte trafficking during pulmonary inflammatory disease in mice

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    Takeshita, W.M.; Gushiken, V.O.; Ferreira-Duarte, A.P.; Pinheiro-Torres, A.S.; Roncalho-Buck, I.A. [Department of Biology and Physiology, Faculty of Medicine of Jundiai (FMJ), Jundiai, SP (Brazil); Squebola-Cola, D.M.; Mello, G.C.; Anhê, G.F.; Antunes, E. [Department of Pharmacology, Faculty of Medical Sciences, University of Campinas (UNICAMP), Campinas, SP (Brazil); DeSouza, I.A., E-mail: ivanidesouza@uol.com.br [Department of Biology and Physiology, Faculty of Medicine of Jundiai (FMJ), Jundiai, SP (Brazil)

    2015-09-15

    Pulmonary neutrophil infiltration produced by Staphylococcal enterotoxin A (SEA) airway exposure is accompanied by marked granulocyte accumulation in bone marrow (BM). Therefore, the aim of this study was to investigate the mechanisms of BM cell accumulation, and trafficking to circulating blood and lung tissue after SEA airway exposure. Male BALB/C mice were intranasally exposed to SEA (1 μg), and at 4, 12 and 24 h thereafter, BM, circulating blood, bronchoalveolar lavage (BAL) fluid and lung tissue were collected. Adhesion of BM granulocytes and flow cytometry for MAC-1, LFA1-α and VLA-4 and cytokine and/or chemokine levels were assayed after SEA-airway exposure. Prior exposure to SEA promoted a marked PMN influx to BAL and lung tissue, which was accompanied by increased counts of immature and/or mature neutrophils and eosinophils in BM, along with blood neutrophilia. Airway exposure to SEA enhanced BM neutrophil MAC-1 expression, and adhesion to VCAM-1 and/or ICAM-1-coated plates. Elevated levels of GM-CSF, G-CSF, INF-γ, TNF-α, KC/CXCL-1 and SDF-1α were detected in BM after SEA exposure. SEA exposure increased production of eosinopoietic cytokines (eotaxin and IL-5) and BM eosinophil VLA-4 expression, but it failed to affect eosinophil adhesion to VCAM-1 and ICAM-1. In conclusion, BM neutrophil accumulation after SEA exposure takes place by integrated action of cytokines and/or chemokines, enhancing the adhesive responses of BM neutrophils and its trafficking to lung tissues, leading to acute lung injury. BM eosinophil accumulation in SEA-induced acute lung injury may occur via increased eosinopoietic cytokines and VLA-4 expression. - Highlights: • Airway exposure to SEA causes acute lung inflammation. • SEA induces accumulation of bone marrow (BM) in immature and mature neutrophils. • SEA increases BM granulocyte or BM PMN adhesion to ICAM-1 and VCAM-1, and MAC-1 expression. • SEA induces BM elevations of CXCL-1, INF-γ, TNF-α, GM-CSF, G-CSF and

  4. Rhinosinusitis derived Staphylococcal enterotoxin B plays a possible role in pathogenesis of food allergy

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    He Shao-Heng

    2006-08-01

    Full Text Available Abstract Background Staphylococcal enterotoxin B (SEB is a potent immunomodulator and implicated with pathogenesis of inflammatory diseases mediated by Th1 or Th2 dominant immune responses. The objective of this study is to determine a possible association between rhinosinusitis derived SEB and pathogenesis of food allergy (FA. Methods The study included chronic rhinosinusitis (CRS patients with FA (N = 46 or without FA (N = 33. Controls included FA patients without CRS (N = 26 and healthy volunteers (N = 25. In CRS patients, we assessed the parameters associated with FA including prick skin test (PST reactivity to food allergens, serum levels of allergen-specific IgE and cytokines (IL-4, IL-13, IFN-Î3, and the number/reactivity of food-allergen specific Th1/Th2 cells in the peripheral blood before and 2 months after sinus surgery. Changes of these parameters were evaluated in comparison with changes in SEB concentration in the sinus lavage and stool samples and also in vitro reactivity to SEB. In CRS patients with FA, we also assessed changes in reactivity to oral challenge of offending food before and after sinus surgery. Results Two months following sinus surgery, we observed statistically significant reduction in PST and oral challenge reactivity in CRS patients with FA in parallel to decrease in serum levels of Th2 cytokines (IL-4 and IL-13 and allergen specific IgE. Improvement of reactivity to food allergens was positively associated with decline in SEB concentrations in the sinus lavage and stool samples. In vitro study results also indicated a role of SEB in aggravation of Th2 skewed responses to food allergens. Such changes were not observed in CRS-non FA patients or control FA patients. Conclusion The rhinosinusitis derived SEB plays a certain role in the pathogenesis of FA by augmenting and/or maintaining polarized Th2 responses. Removal of SEB-producing pathogens from the rhinosinuses may be beneficial for attenuating the FA

  5. A cfr-positive clinical staphylococcal isolate from India with multiple mechanisms of linezolid-resistance

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    Vineeth Rajan

    2014-01-01

    Full Text Available Background & objectives: Linezolid, a member of the oxazolidinone class of antibiotics, has been an effective therapeutic option to treat severe infections caused by multidrug resistant Gram positive bacteria. Emergence of linezolid resistant clinical strains is a serious issue in the healthcare settings worldwide. We report here the molecular characterization of a linezolid resistant clinical isolate of Staphylococcus haemolyticus from India. Methods: The species of the clinical isolate was identified by 16S rRNA gene sequencing. The minimum inhibitory concentrations (MICs of linezolid, clindamycin, chloramphenicol and oxacillin were determined by E-test method. To elucidate the mechanism of linezolid-resistance, presence of cfr gene (chloramphenicol florfenicol resistance and mutations in 23S rRNA and ribosomal proteins (L3, L4 and L22 were investigated. Staphylococcal Cassette Chromosome mec (SCCmec typing was performed by multiplex PCR. Results: The study documented a rare clinical S. haemolyticus strain with three independent mechanisms of linezolid-resistance. The strain carried cfr gene, the only known transmissible mechanism of linezolid-resistance. The strain also possessed resistance-conferring mutations such as G 2576 T in domain V of 23S rRNA gene and Met 156 Thr in L3 ribosomal protein. The other ribosomal proteins (L4 and L22 did not exhibit mutations accountable for linezolid-resistance. Restriction digestion by NheI revealed that all the alleles of 23S rRNA gene were mutated. The isolate showed elevated MIC values (>256 ΅g ml -[1] of linezolid, clindamycin, chloramphenicol and oxacillin. Methicillin resistance was conferred by type I SCCmec element. The strain also harboured lsa(B gene which encodes an ABC transporter that can efflux clindamycin. Interpretation & conclusions: The present study reports the first clinical strain from India with transmissible and multiple mechanisms of linezolid-resistance. Judicious use of

  6. Epidemiology and prognosis of coagulase-negative staphylococcal endocarditis: impact of vancomycin minimum inhibitory concentration.

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    García de la Mària, Cristina; Cervera, Carlos; Pericàs, Juan M; Castañeda, Ximena; Armero, Yolanda; Soy, Dolors; Almela, Manel; Ninot, Salvador; Falces, Carlos; Mestres, Carlos A; Gatell, Jose M; Moreno, Asuncion; Marco, Francesc; Miró, José M

    2015-01-01

    This study describes coagulase-negative staphylococcal (CoNS) infective endocarditis (IE) epidemiology at our institution, the antibiotic susceptibility profile, and the influence of vancomycin minimum inhibitory concentration (MIC) on patient outcomes. One hundred and three adults with definite IE admitted to an 850-bed tertiary care hospital in Barcelona from 1995-2008 were prospectively included in the cohort. We observed that CoNS IE was an important cause of community-acquired and healthcare-associated IE; one-third of patients involved native valves. Staphylococcus epidermidis was the most frequent species, methicillin-resistant in 52% of patients. CoNS frozen isolates were available in 88 patients. Vancomycin MICs of 2.0 μg/mL were common; almost all cases were found among S. epidermidis isolates and did not increase over time. Eighty-five patients were treated either with cloxacillin or vancomycin: 38 patients (Group 1) were treated with cloxacillin, and 47 received vancomycin; of these 47, 27 had CoNS isolates with a vancomycin MIC <2.0 μg/mL (Group 2), 20 had isolates with a vancomycin MIC ≥ 2.0 μg/mL (Group 3). One-year mortality was 21%, 48%, and 65% in Groups 1, 2, and 3, respectively (P = 0.003). After adjusting for confounders and taking Group 2 as a reference, methicillin-susceptibility was associated with lower 1-year mortality (OR 0.12, 95% CI 0.02-0.55), and vancomycin MIC ≥ 2.0 μg/mL showed a trend to higher 1-year mortality (OR 3.7, 95% CI 0.9-15.2; P=0.069). Other independent variables associated with 1-year mortality were heart failure (OR 6.2, 95% CI 1.5-25.2) and pacemaker lead IE (OR 0.1, 95%CI 0.02-0.51). In conclusion, methicillin-resistant S.epidermidis was the leading cause of CoNS IE, and patients receiving vancomycin had higher mortality rates than those receiving cloxacillin; mortality was higher among patients having isolates with vancomycin MICs ≥ 2.0 μg/mL.

  7. A scabies mite serpin interferes with complement-mediated neutrophil functions and promotes staphylococcal growth.

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    Pearl M Swe

    2014-06-01

    Full Text Available BACKGROUND: Scabies is a contagious skin disease caused by the parasitic mite Sarcoptes scabiei. The disease is highly prevalent worldwide and known to predispose to secondary bacterial infections, in particular by Streptococcus pyogenes and Staphylococcus aureus. Reports of scabies patients co-infected with methicillin resistant S. aureus (MRSA pose a major concern for serious down-stream complications. We previously reported that a range of complement inhibitors secreted by the mites promoted the growth of S. pyogenes. Here, we show that a recently characterized mite serine protease inhibitor (SMSB4 inhibits the complement-mediated blood killing of S. aureus. METHODOLOGY/PRINCIPAL FINDINGS: Blood killing of S. aureus was measured in whole blood bactericidal assays, counting viable bacteria recovered after treatment in fresh blood containing active complement and phagocytes, treated with recombinant SMSB4. SMSB4 inhibited the blood killing of various strains of S. aureus including methicillin-resistant and methicillin-sensitive isolates. Staphylococcal growth was promoted in a dose-dependent manner. We investigated the effect of SMSB4 on the complement-mediated neutrophil functions, namely phagocytosis, opsonization and anaphylatoxin release, by flow cytometry and in enzyme linked immuno sorbent assays (ELISA. SMSB4 reduced phagocytosis of S. aureus by neutrophils. It inhibited the deposition of C3b, C4b and properdin on the bacteria surface, but did not affect the depositions of C1q and MBL. SMSB4 also inhibited C5 cleavage as indicated by a reduced C5b-9 deposition. CONCLUSIONS/SIGNIFICANCE: We postulate that SMSB4 interferes with the activation of all three complement pathways by reducing the amount of C3 convertase formed. We conclude that SMSB4 interferes with the complement-dependent killing function of neutrophils, thereby reducing opsonization, phagocytosis and further recruitment of neutrophils to the site of infection. As a

  8. Staphylococcus aureus soft tissue infection may increase the risk of subsequent staphylococcal soft tissue infections.

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    Bouvet, Cindy; Gjoni, Shpresa; Zenelaj, Besa; Lipsky, Benjamin A; Hakko, Elif; Uçkay, Ilker

    2017-07-01

    Staphylococcus aureus is the most common cause of soft tissue infections. It is unknown, however, if a patient who has had such an infection is at greater risk for future soft tissue infections with S. aureus. We conducted an epidemiological survey of adult patients hospitalized in the only public hospital in Geneva for treatment (usually combined surgical and medical) of a soft tissue infection caused by S. aureus. By reviewing nursing and medical records from the emergency department and hospital wards, we assessed whether or not they developed any other soft tissue infections (excluding a recurrence) after or before the index one. Among 1023 index episodes of soft tissue infections, 670 (65%) were caused by S. aureus, of which 47 were caused by methicillin-resistant strains (30 healthcare-associated and 17 community-acquired). The patients' median age was 51 years and 334 (34%) were immune-compromised. The median time span between the patient's first and last consultation (for any reason) in our hospital was 21.4 years (interquartile range, 10-30 years). In addition to their index infection, 124 patients (12%) developed a new nosocomial or community-acquired soft tissue infection. Among the index cases with an S. aureus infection, 92 (14%) had another soft tissue infection, compared to 32 (9%) who had a non-staphylococcal index infection (Pearson-χ(2)-test; p=0.03). Similarly, patients with an index S. aureus infection, compared to those with a non-S. aureus infection, had a higher rate of another soft tissue infection caused by S. aureus (χ(2)-test; pS. aureus shows a high association to further S. aureus soft tissue infections (logistic regression; odds ratio 2.5, 95% confidence interval 1.4-4.6). Among adult patients hospitalised for a soft tissue infection, those infected with S. aureus (compared with other pathogens) may be at higher risk of a subsequent soft tissue infection, particularly with S. aureus. Copyright © 2017 The Author(s). Published by

  9. Staphylococcal toxic shock syndrome 2000-2006: epidemiology, clinical features, and molecular characteristics.

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    Aaron S DeVries

    Full Text Available INTRODUCTION: Circulating strains of Staphylococcus aureus (SA have changed in the last 30 years including the emergence of community-associated methicillin-resistant SA (MRSA. A report suggested staphylococcal toxic shock syndrome (TSS was increasing over 2000-2003. The last population-based assessment of TSS was 1986. METHODS: Population-based active surveillance for TSS meeting the CDC definition using ICD-9 codes was conducted in the Minneapolis-St. Paul area (population 2,642,056 from 2000-2006. Medical records of potential cases were reviewed for case criteria, antimicrobial susceptibility, risk factors, and outcome. Superantigen PCR testing and PFGE were performed on available isolates from probable and confirmed cases. RESULTS: Of 7,491 hospitalizations that received one of the ICD-9 study codes, 61 TSS cases (33 menstrual, 28 non-menstrual were identified. The average annual incidence per 100,000 of all, menstrual, and non-menstrual TSS was 0.52 (95% CI, 0.32-0.77, 0.69 (0.39-1.16, and 0.32 (0.12-0.67, respectively. Women 13-24 years had the highest incidence at 1.41 (0.63-2.61. No increase in incidence was observed from 2000-2006. MRSA was isolated in 1 menstrual and 3 non-menstrual cases (7% of TSS cases; 1 isolate was USA400. The superantigen gene tst-1 was identified in 20 (80% of isolates and was more common in menstrual compared to non-menstrual isolates (89% vs. 50%, p = 0.07. Superantigen genes sea, seb and sec were found more frequently among non-menstrual compared to menstrual isolates [100% vs 25% (p = 0.4, 60% vs 0% (p<0.01, and 25% vs 13% (p = 0.5, respectively]. DISCUSSION: TSS incidence remained stable across our surveillance period of 2000-2006 and compared to past population-based estimates in the 1980s. MRSA accounted for a small percentage of TSS cases. tst-1 continues to be the superantigen associated with the majority of menstrual cases. The CDC case definition identifies the most severe cases and has

  10. Systematic survey on the prevalence of genes coding for staphylococcal enterotoxins SElM, SElO, and SElN

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    Becker, Karsten; Friedrich, Alexander W; Peters, Georg; von Eiff, Christof

    2004-01-01

    Staphylococcus aureus remains a leading cause of food-poisoning with substantial impact on public health. Using a multiplex polymerase chain reaction-DNA enzyme immunoassay (PCR-DEIA), we studied the presence of genes encoding staphylococcal enterotoxin-like (SEl) superantigens sem, sen, and seo, as

  11. Reprint of "Identification of staphylococcal species based on variations in protein sequences (mass spectrometry) and DNA sequence (sodA microarray)".

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    Kooken, Jennifer; Fox, Karen; Fox, Alvin; Altomare, Diego; Creek, Kim; Wunschel, David; Pajares-Merino, Sara; Martínez-Ballesteros, Ilargi; Garaizar, Javier; Oyarzabal, Omar; Samadpour, Mansour

    2014-01-01

    This report is among the first using sequence variation in newly discovered protein markers for staphylococcal (or indeed any other bacterial) speciation. Variation, at the DNA sequence level, in the sodA gene (commonly used for staphylococcal speciation) provided excellent correlation. Relatedness among strains was also assessed using protein profiling using microcapillary electrophoresis and pulsed field electrophoresis. A total of 64 strains were analyzed including reference strains representing the 11 staphylococcal species most commonly isolated from man (Staphylococcus aureus and 10 coagulase negative species [CoNS]). Matrix assisted time of flight ionization/ionization mass spectrometry (MALDI TOF MS) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC ESI MS/MS) were used for peptide analysis of proteins isolated from gel bands. Comparison of experimental spectra of unknowns versus spectra of peptides derived from reference strains allowed bacterial identification after MALDI TOF MS analysis. After LC-MS/MS analysis of gel bands bacterial speciation was performed by comparing experimental spectra versus virtual spectra using the software X!Tandem. Finally LC-MS/MS was performed on whole proteomes and data analysis also employing X!tandem. Aconitate hydratase and oxoglutarate dehydrogenase served as marker proteins on focused analysis after gel separation. Alternatively on full proteomics analysis elongation factor Tu generally provided the highest confidence in staphylococcal speciation.

  12. Noncontiguous Finished Genome Sequence of Staphylococcus aureus KLT6, a Staphylococcal Enterotoxin B-Positive Strain Involved in a Food Poisoning Outbreak in Switzerland

    Science.gov (United States)

    Tobes, Raquel; Manrique, Marina; Brozynska, Marta; Stephan, Roger; Pareja, Eduardo

    2013-01-01

    We present the first complete genome sequence of a Staphylococcus aureus strain assigned to clonal complex 12. The strain was isolated in a food poisoning outbreak due to contaminated potato salad in Switzerland in 2009, and it produces staphylococcal enterotoxin B. PMID:23704175

  13. Differential interaction of the staphylococcal toxins panton-valentine leukocidin and γ-hemolysin CB with human C5a receptors

    NARCIS (Netherlands)

    Spaan, András N.; Schiepers, Ariën; De Haas, Carla J C; Van Hooijdonk, Davy D J J; Badiou, Cédric; Contamin, Hugues; Vandenesch, François; Lina, Gérard; Gerard, Norma P.; Gerard, Craig; Van Kessel, Kok P M; Henry, Thomas; Van Strijp, Jos A G

    2015-01-01

    Staphylococcus aureus is well adapted to the human host. Evasion of the host phagocyte response is critical for successful infection. The staphylococcal bicomponent pore-forming toxins Panton-Valentine leukocidin LukSF-PV (PVL) and γ-hemolysin CB (HlgCB) target human phagocytes through interaction w

  14. A Seven-Year Survey of Management of Coagulase-Negative Staphylococcal Sepsis in the Neonatal Intensive Care Unit : Vancomycin May Not Be Necessary as Empiric Therapy

    NARCIS (Netherlands)

    Hemels, Marieke A. C.; van den Hoogen, Agnes; Verboon-Maciolek, Malgorzata A.; Fleer, Andre; Krediet, Tannette G.

    2011-01-01

    Background: The typical empiric therapy for coagulase-negative staphylococcal (CONS) sepsis includes vancomycin. In our neonatal intensive care unit, we have consistently avoided the use of vancomycin to treat CONS sepsis, except for specific cases, and have used instead cefazolin as empiric agent.

  15. Modelling staphylococcal pneumonia in a human 3D lung tissue model system delineates toxin-mediated pathology

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    Srikanth Mairpady Shambat

    2015-11-01

    Full Text Available Staphylococcus aureus necrotizing pneumonia is recognized as a toxin-mediated disease, yet the tissue-destructive events remain elusive, partly as a result of lack of mechanistic studies in human lung tissue. In this study, a three-dimensional (3D tissue model composed of human lung epithelial cells and fibroblasts was used to delineate the role of specific staphylococcal exotoxins in tissue pathology associated with severe pneumonia. To this end, the models were exposed to the mixture of exotoxins produced by S. aureus strains isolated from patients with varying severity of lung infection, namely necrotizing pneumonia or lung empyema, or to purified toxins. The necrotizing pneumonia strains secreted high levels of α-toxin and Panton-Valentine leukocidin (PVL, and triggered high cytotoxicity, inflammation, necrosis and loss of E-cadherin from the lung epithelium. In contrast, the lung empyema strain produced moderate levels of PVL, but negligible amounts of α-toxin, and triggered limited tissue damage. α-toxin had a direct damaging effect on the epithelium, as verified using toxin-deficient mutants and pure α-toxin. Moreover, PVL contributed to pathology through the lysis of neutrophils. A combination of α-toxin and PVL resulted in the most severe epithelial injury. In addition, toxin-induced release of pro-inflammatory mediators from lung tissue models resulted in enhanced neutrophil migration. Using a collection of 31 strains from patients with staphylococcal pneumonia revealed that strains producing high levels of α-toxin and PVL were cytotoxic and associated with fatal outcome. Also, the strains that produced the highest toxin levels induced significantly greater epithelial disruption. Of importance, toxin-mediated lung epithelium destruction could be inhibited by polyspecific intravenous immunoglobulin containing antibodies against α-toxin and PVL. This study introduces a novel model system for study of staphylococcal pneumonia in a

  16. Modelling staphylococcal pneumonia in a human 3D lung tissue model system delineates toxin-mediated pathology.

    Science.gov (United States)

    Mairpady Shambat, Srikanth; Chen, Puran; Nguyen Hoang, Anh Thu; Bergsten, Helena; Vandenesch, Francois; Siemens, Nikolai; Lina, Gerard; Monk, Ian R; Foster, Timothy J; Arakere, Gayathri; Svensson, Mattias; Norrby-Teglund, Anna

    2015-11-01

    Staphylococcus aureus necrotizing pneumonia is recognized as a toxin-mediated disease, yet the tissue-destructive events remain elusive, partly as a result of lack of mechanistic studies in human lung tissue. In this study, a three-dimensional (3D) tissue model composed of human lung epithelial cells and fibroblasts was used to delineate the role of specific staphylococcal exotoxins in tissue pathology associated with severe pneumonia. To this end, the models were exposed to the mixture of exotoxins produced by S. aureus strains isolated from patients with varying severity of lung infection, namely necrotizing pneumonia or lung empyema, or to purified toxins. The necrotizing pneumonia strains secreted high levels of α-toxin and Panton-Valentine leukocidin (PVL), and triggered high cytotoxicity, inflammation, necrosis and loss of E-cadherin from the lung epithelium. In contrast, the lung empyema strain produced moderate levels of PVL, but negligible amounts of α-toxin, and triggered limited tissue damage. α-toxin had a direct damaging effect on the epithelium, as verified using toxin-deficient mutants and pure α-toxin. Moreover, PVL contributed to pathology through the lysis of neutrophils. A combination of α-toxin and PVL resulted in the most severe epithelial injury. In addition, toxin-induced release of pro-inflammatory mediators from lung tissue models resulted in enhanced neutrophil migration. Using a collection of 31 strains from patients with staphylococcal pneumonia revealed that strains producing high levels of α-toxin and PVL were cytotoxic and associated with fatal outcome. Also, the strains that produced the highest toxin levels induced significantly greater epithelial disruption. Of importance, toxin-mediated lung epithelium destruction could be inhibited by polyspecific intravenous immunoglobulin containing antibodies against α-toxin and PVL. This study introduces a novel model system for study of staphylococcal pneumonia in a human setting. The

  17. Efeito do escaldamento nas propriedades tecnológicas e reológicas da massa e do pão de queijo Effect of scalding on technological and rheological properties of cheese bread dough and cheese bread

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    Antônio Vitor Machado

    2010-04-01

    Full Text Available O pão de queijo é um produto da culinária brasileira de origem mineira, amplamente consumido e conhecido até mesmo internacionalmente. Apesar de ser um produto largamente consumido no mercado, não possui padronização de produção, e qualidade bem definidos. Trabalhos a seu respeito são raros, faltando informações sobre o produto e seus padrões de qualidade, tornando-se necessário um melhor conhecimento dos efeitos da operação de escaldamento nas propriedades reológicas e tecnológicas da massa e do pão de queijo, como também dos ingredientes utilizados nas etapas de sua produção. Este trabalho foi desenvolvido no Departamento de Ciência dos Alimentos da Universidade Federal de Lavras, Minas Gerais e um dos objetivos foi estudar o comportamento reológico da massa de pão de queijo em função dos métodos de preparo empregado, com ou sem escaldamento, utilizando leite ou água na formulação. Outro, foi avaliar o comportamento da massa durante as etapas subsequentes de adição e incorporação de ovo e queijo na massa. De acordo com os resultados, o escaldamento demonstrou ser de grande importância na produção de massa de pão de queijo influenciando diretamente na qualidade física e de textura.Cheese bread is a Brazilian cuisine specialty originated in the Minas Gerais region. It is widely produced and consumed, and even known internationally. In spite of being widely consumed, there is no production standard, identity and defined quality. Research on the subject is rare, lacking information on the product and its quality patterns, making it necessary to have a better understanding of the effect of scalding on the rheological properties and technology of the cheese bread dough, as well as of the ingredients in the production stages. This work was carried out in the Department of Food Science at the Federal University of Lavras, Minas Gerais and one of the objectives was to study the rheological behavior of the cheese

  18. Characterization of a Novel Composite Staphylococcal Cassette Chromosome mec in Methicillin-Resistant Staphylococcus pseudintermedius from Thailand.

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    Chanchaithong, Pattrarat; Prapasarakul, Nuvee; Perreten, Vincent; Schwendener, Sybille

    2016-02-01

    A novel staphylococcal cassette chromosome mec (SCCmec) composite island (SCCmecAI16-SCCczrAI16-CI) was identified in Staphylococcus pseudintermedius. Four integration site sequences for SCC subdivided the 60,734-bp island into 41,232-bp SCCmecAI16, 19,400-bp SCCczrAI16, and 102-bp SCC-likeAI16 elements. SCCmecAI16 represents a new combination of ccrA1B3 genes with a class A mec complex. SCCczrAI16 contains ccrA1B6 and genes related to restriction modification and heavy metal resistance. SCCmecAI16-SCCczrAI16-CI was found in methicillin-resistant S. pseudintermedius sequence type 112 (ST112) and ST111 isolated from dogs and veterinarians in Thailand.

  19. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Science.gov (United States)

    Hill, F. S.; Cox, A. B.; Salmon, Y. L.; Cantu, A. O.; Lucas, J. N.

    1994-01-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 microgram/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) than that produced by PHA (MI < 0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture.

  20. Structure of Staphylococcal Enterotoxin E in Complex with TCR Defines the Role of TCR Loop Positioning in Superantigen Recognition.

    Directory of Open Access Journals (Sweden)

    Karin E J Rödström

    Full Text Available T cells are crucial players in cell-mediated immunity. The specificity of their receptor, the T cell receptor (TCR, is central for the immune system to distinguish foreign from host antigens. Superantigens are bacterial toxins capable of inducing a toxic immune response by cross-linking the TCR and the major histocompatibility complex (MHC class II and circumventing the antigen specificity. Here, we present the structure of staphylococcal enterotoxin E (SEE in complex with a human T cell receptor, as well as the unligated T cell receptor structure. There are clear structural changes in the TCR loops upon superantigen binding. In particular, the HV4 loop moves to circumvent steric clashes upon complex formation. In addition, a predicted ternary model of SEE in complex with both TCR and MHC class II displays intermolecular contacts between the TCR α-chain and the MHC, suggesting that the TCR α-chain is of importance for complex formation.

  1. Staphylococcal diaphyseal subacute osteomyelitis of the ulna in a child: an unusual cause of post-traumatic forearm swelling.

    LENUS (Irish Health Repository)

    Kearns, S R

    2004-01-01

    Paediatric subacute osteomyelitis (SAO) presents a diagnostic and therapeutic challenge to clinicians. Typically located in the metaphysis of long bones, diaphyseal SAO of the upper limb is rare. We present the case of a three-year-old girl referred to our fracture clinic as an occult fracture following trauma to her forearm with normal initial radiographs. Follow-up radiographs one week later showed cortical erosion of the distal ulna, while a subsequent MRI scan showed soft tissue swelling with an area of high signal in the distal ulna. A limited biopsy diagnosed staphylococcal subacute osteomyelitis of the ulna. The patient responded to high dose antibiotic therapy and made a full recovery. We present this case to highlight the high index of suspicion required to diagnose and appropriately manage this insidious condition, which may easily be confused with any number of benign and malignant bony lesions and provide a review of the relevant literature.

  2. Human Tudor staphylococcal nuclease (Tudor-SN) protein modulates the kinetics of AGTR1-3'UTR granule formation.

    Science.gov (United States)

    Gao, Xingjie; Shi, Xuebin; Fu, Xue; Ge, Lin; Zhang, Yi; Su, Chao; Yang, Xi; Silvennoinen, Olli; Yao, Zhi; He, Jinyan; Wei, Minxin; Yang, Jie

    2014-06-13

    Human Tudor staphylococcal nuclease (Tudor-SN) interacts with the G3BP protein and is recruited into stress granules (SGs), the main type of discrete RNA-containing cytoplasmic foci structure that is formed under stress conditions. Here, we further demonstrate that Tudor-SN binds and co-localizes with AGTR1-3'UTR (3'-untranslated region of angiotensin II receptor, type 1 mRNA) into SG. Tudor-SN plays an important role in the assembly of AGTR1-3'UTR granules. Moreover, endogenous Tudor-SN knockdown can decrease the recovery kinetics of AGTR1-3'UTR granules. Collectively, our data indicate that Tudor-SN modulates the kinetics of AGTR1-3'UTR granule formation, which provides an additional biological role of Tudor-SN in RNA metabolism during stress. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  3. Clostridium difficile bacteremia and meningitis as a complication of prolonged cephalosporin therapy in a case of staphylococcal pyogenic arthritis

    Institute of Scientific and Technical Information of China (English)

    Abhrajit Ganguly; Saibal Das; Jayanta Kumar Dey; Somnath Mondal

    2012-01-01

    With increasing incidence of Clostridium difficile (C. difficile) associated diarrhea and pseudomembranous colitis, several extra-intestinal manifestations of the organism have been unmasked which include-bacteremia, brain abscess, pericarditis etc. We report a rare and interesting case of C. difficile bacteremia and subsequent meningitis in a 10 year old child. The child was immune competent, which further raises the question about the virulent possibilities of the organism and its implications in the near future. The condition resulted from a prolonged treatment with intravenous (I.V.) cefotaxime for staphylococcal pyogenic arthritis. The child recovered from the septic arthritis but on the 7th day post-admission developed features of bacteremia. The child was later treated with intravenous metronidazole and vancomycin and he was discharged on the 21st day post-admission. No recurrence of symptoms was noted.

  4. Clostridium difficile bacteremia and meningitis as a complication of prolonged cephalosporin therapy in a case of staphylococcal pyogenic arthritis

    Directory of Open Access Journals (Sweden)

    Abhrajit Ganguly

    2012-01-01

    Full Text Available With increasing incidence of Clostridium difficile (C. difficile associated diarrhea and pseudomembranous colitis, several extra-intestinal manifestations of the organism have been unmasked which include-bacteremia, brain abscess, pericarditis etc. We report a rare and interesting case of C. difficile bacteremia and subsequent meningitis in a 10 year old child. The child was immune competent, which further raises the question about the virulent possibilities of the organism and its implications in the near future. The condition resulted from a prolonged treatment with intravenous (I.V. cefotaxime for staphylococcal pyogenic arthritis. The child recovered from the septic arthritis but on the 7th day post-admission developed features of bacteremia. The child was later treated with intravenous metronidazole and vancomycin and he was discharged on the 21st day post-admission. No recurrence of symptoms was noted.

  5. Conductometric immunosensors for the detection of staphylococcal enterotoxin B based bio-electrocalytic reaction on micro-comb electrodes.

    Science.gov (United States)

    Chen, Zai-Gang

    2008-06-01

    Staphylococcal enterotoxin B (SEB) is one of many toxins produced by the Gram-positive bacterium Staphylococcal aureus. While SEB is known as the causative agent of certain food poisonings it is also considered abiological select agent. Thus, rapid and accurate identification of SEB during either surveillance or in response to a biothreat is critical to the mitigation of the suspect agent. This report presents a new conductometric immune-biosensor for the detection of SEB based on immobilization of horseradish peroxidase (HRP)-labeled SEB antibody (HRP-anti-SEB) onto nanogold/chitosan-multiwalled carbon nanotube (Au/CTS-MWNT)-functionalized biorecognition interface. The formation of the antibody-antigen complex by a simple one-step immunoreaction between the immobilized HRP-anti-SEB and SEB in sample solution introduced a barrier of electrical communication between the immobilized HRP and the base surface, thus local conductivity variations could be evaluated by the bio-electrocatalytic reaction of HRP in 0.02 M PBS (pH 6.8) containing 0.15 mM H(2)O(2), 0.06 M KI and 0.1 M NaCl. Under optimal conditions, the proposed immune-biosensor exhibited a good conductometric response relative to SEB concentration in a linear range from 0.5 to 83.5 ng/ml with a correlation coefficient of 0.998. The developed immune-biosensor showed an acceptable accuracy, reproducibility and stability. Milk samples spiked with various concentrations of SEB gave an average of 116% recovery of the toxin.

  6. Staphylococcal phenotypes induced by naturally occurring and synthetic membrane-interactive polyphenolic β-lactam resistance modifiers.

    Directory of Open Access Journals (Sweden)

    Lucia Palacios

    Full Text Available Galloyl catechins, in particular (--epicatechin gallate (ECg, have the capacity to abrogate β-lactam resistance in methicillin-resistant strains of Staphylococcus aureus (MRSA; they also prevent biofilm formation, reduce the secretion of a large proportion of the exoproteome and induce profound changes to cell morphology. Current evidence suggests that these reversible phenotypic traits result from their intercalation into the bacterial cytoplasmic membrane. We have endeavoured to potentiate the capacity of ECg to modify the MRSA phenotype by stepwise removal of hydroxyl groups from the B-ring pharmacophore and the A:C fused ring system of the naturally occurring molecule. ECg binds rapidly to the membrane, inducing up-regulation of genes responsible for protection against cell wall stress and maintenance of membrane integrity and function. Studies with artificial membranes modelled on the lipid composition of the staphylococcal bilayer indicated that ECg adopts a position deep within the lipid palisade, eliciting major alterations in the thermotropic behaviour of the bilayer. The non-galloylated homolog (--epicatechin enhanced ECg-mediated effects by facilitating entry of ECg molecules into the membrane. ECg analogs with unnatural B-ring hydroxylation patterns induced higher levels of gene expression and more profound changes to MRSA membrane fluidity than ECg but adopted a more superficial location within the bilayer. ECg possessed a high affinity for the positively charged staphylococcal membrane and induced changes to the biophysical properties of the bilayer that are likely to account for its capacity to disperse the cell wall biosynthetic machinery responsible for β-lactam resistance. The ability to enhance these properties by chemical modification of ECg raises the possibility that more potent analogs could be developed for clinical evaluation.

  7. 烫伤大鼠结肠动力基本功能单位细胞损害和内质网应激研究%Study on endoplasmic reticulum stress and injury to basic function unit of colonic motility in rats with scald injury

    Institute of Scientific and Technical Information of China (English)

    李颖; 郭力

    2009-01-01

    路致细胞损伤可能有关.%Objective To observe expressions of glucose-regulated protein (GRF78) and caspase-12 in nervous system-interstitial cells of Cajal (ICC) in smooth muscle in colonic wall in rats with scald inju-ry, as well as their relevant ultrastructural changes, so as to probe the possible mechanisms of dynamic dam-age in murine colon after a scald injury. Methods Fifty healthy Sprague-Dawley rats were randomly di-vided into scald (n =40) and control (n = 10) groups. Rats in scald group were inflicted with 30% TBSA full-thickness scald, and received an intraperitoneally injection of Ringer lactate solution (50 mg/kg) for re-suscitation, while those in control group had similar treatment with the exception of scald. Rats in control group and scald group were sacrificed at 3, 6, 12, 24 post scald hour ( PSH, 10 rats at each time point) for collection of 4 cm of colonic tissue, 5 cm proximal to the cecum. A segment of colonic wall, 1 cm in length, was obtained from the middle of the harvested segment of colon, and it was fixed with 3% glutaraldebyde or 10% formaldehyde. The samples fixed with glutaraldehyde were used to observe uhrastructural alterations under transmission electron microscope, while that with formaldehyde were used to observe expressions of GRP78 and caspase-12 in colonic wall by immunohistochemical assay. Results The colonic smooth mus-cle cells of rats in control group showed regular arrangment, their organelles were abundant, nucleus central-ly located, euchromatin distributed evenly with more abundant mitochondrial cristae and less smooth endo-plasmic reticulum, neuronal organelles were abundant in intermuscular plexus, and ICC could be seen in the neighborhood of neurons. The colonic smooth muscle cells appeared in irregular and disordered manner in scald group, perinuclear space was widened, intercellular vacuoles were observed, mitochondria showed vac-uolation degeneration with dissolved and condensed cristae, rough endoplasmic

  8. Electro-acupuncture at Zusanli (足三里) alleviates pathologic injury induced by intestinal proinflamatory factors in scalded rats%电针足三里穴减轻烫伤大鼠小肠促炎细胞因子引起的病理损害

    Institute of Scientific and Technical Information of China (English)

    胡森; 王磊; 周国勇

    2011-01-01

    Objective To investigate the inhibitory effects of electro-acupuncture(EA) at Zusanli (足三里) on small intestinal proinflammatory factors and tissue insult in rats with scald injury and their relationship to cholinergic nervous pathway. Methods Forty Wistar rats were randomly divided into four groups (each,n= 10): scald + EA group (S/EA group), scald + shame EA group (S/SEA group), vagotomy + scald +shame EA group (VA/S/SEA group) and vagotomy +scald +EA group (VA/S/EA group). The rat's back was subjected to boiling water (96 ℃) for 15 seconds to produce a 35 % full thickness scald injury of total body surface area (TBSA). Twenty minutes after scald, bilateral Zusanli points were persistently electroacupunctured with constant voltage (2 -3 mA, 2 - 100 Hz for 30 minutes ). The shame EA group was performed at a point 0. 5 cm lateral to the Zusanli point with the same frequency and intensity of stimulation for 30 minutes. Abdominal vagotomy was performed in rats in VA/S/SEA and VA/S/EA groups before scald.All the rats were sacrificed 6 hours after scald, and the specimens of jejunum were harvested to evaluate the activities of tumor necrosis factor-α (TNF-α), nitric oxide ( NO ), nitric oxide synthase ( NOS ),myeloperoxidase (MPO), diamine oxidase (DAO) and assessment of the rate of water content (ratio of dry to wet weight). Results Compared to the S/EA group, the levels of TNF-α and NO, activities of NOS and MPO and the rates of water content in jejunum were significantly higher and the activity of DAO was obviously lower in S/SEA, VA/S/SEA and VA/S/EA groups 6 hours after scald (all P<0. 01). Compared to the S/SEA group, the level of TNF-α was markedly higher and of DAO obviously lower in VA/S/SEA and VA/S/EA groups (all P<0. 05); the comparisons of levels of NO、NOS and MPO did not reveal any statistical significant differences (P>0. 05). In regard to the changes of all the above indexes, no statistical differences were seen in VA/S/SEA and VA

  9. Genomic and evolutionary features of the SPI-1 type III secretion system that is present in Xanthomonas albilineans but is not essential for xylem colonization and symptom development of sugarcane leaf scald.

    Science.gov (United States)

    Marguerettaz, Mélanie; Pieretti, Isabelle; Gayral, Philippe; Puig, Jérôme; Brin, Chrystelle; Cociancich, Stéphane; Poussier, Stéphane; Rott, Philippe; Royer, Monique

    2011-02-01

    Xanthomonas albilineans is the causal agent of sugarcane leaf scald. Interestingly, this bacterium, which is not known to be insect or animal associated, possesses a type III secretion system (T3SS) belonging to the injectisome family Salmonella pathogenicity island 1 (SPI-1). The T3SS SPI-1 of X. albilineans shares only low similarity with other available T3SS SPI-1 sequences. Screening of a collection of 128 plant-pathogenic bacteria revealed that this T3SS SPI-1 is present in only two species of Xanthomonas: X. albilineans and X. axonopodis pv. phaseoli. Inoculation of sugarcane with knockout mutants showed that this system is not required by X. albilineans to spread within xylem vessels and to cause disease symptoms. This result was confirmed by the absence of this T3SS SPI-1 in an X. albilineans strain isolated from diseased sugarcane. To investigate the importance of the T3SS SPI-1 during the life cycle of X. albilineans, we analyzed T3SS SPI-1 sequences from 11 strains spanning the genetic diversity of this species. No nonsense mutations or frameshifting indels were observed in any of these strains, suggesting that the T3SS SPI-1 system is maintained within the species X. albilineans. Evolutionary features of T3SS SPI-1 based on phylogenetic, recombination, and selection analyses are discussed in the context of the possible functional importance of T3SS SPI-1 in the ecology of X. albilineans.

  10. Effect of the Cooling System and 1-MCP on the Incidence of Superficial Scald in ‘Granny Smith’ Apples Efecto del Sistema de Enfriamiento y 1-MCP sobre la Incidencia de Escaldado Superficial en Manzanas ‘Granny Smith’

    Directory of Open Access Journals (Sweden)

    Claudia Moggia

    2009-09-01

    Full Text Available A study was carried out to determine the effects of two cooling systems and the application of 1-methylcyclopropene (1-MCP, SmartFreshTM on the incidence of superficial scald in apples (Malus domestica Borkh. cv. Granny Smith. Fruit were collected from a commercial orchard (Colbún, Maule Region, Chile during 2004-2005 season. A completely randomized design was used in a 2 x 2 factorial arrangement, using cooling systems (normal and step-wise cooling and application of 1-MCP (0 and 625 nL L-1 i.a. as the main factors. Fruit from normal cooling were kept at 0 °C throughout the storage period (180 days. Step-wise cooling consisted of storing the fruit at 10 °C for 10 days; 4 °C for the next 20 days and 0 °C for the remaining 150 days. Every month, maturity indices and the chemicals, global antioxidant content (AO, α-farnesene (AF and conjugated trienes (TC, were measured. Incidence of superficial scald was determined after 180 days of storage, plus 10 days at 20 °C . The application of 1-MCP with both cooling systems was capable of maintaining firmness values around 8,2 kg up to the end of the storage period, as well as decreasing the concentration of AF, TC and AO during storage. Incidence of superficial scald on fruit with 1-MCP was 0%, regardless of the type of cooling. Among treatments without 1-MCP, step-wise-cooling was more effective in preventing superficial scald, resulting in 1.3% incidence compared to 75.6% with the normal cooling system. Nevertheless, from the fourth month onwards firmness was lower than that required for export.Se desarrolló un estudio para determinar el efecto de dos sistemas de enfriamiento y la aplicación de 1-metilciclopropeno (1-MCP, SmartFreshTM sobre la incidencia de escaldado superficial en manzanas (Malus domestica Borkh. cv. Granny Smith, colectadas de un huerto comercial (Colbún, Región del Maule, Chile durante la temporada 2004-2005. El ensayo consistió en un diseño completamente al azar con

  11. 胞壁酰二肽诱导烫伤大鼠脓毒症模型的建立%Establishment of a rat model of sepsis induced by muramyl dipeptide after scald burn

    Institute of Scientific and Technical Information of China (English)

    韩毅; 宋学敏; 李建国; 梁辉; 刘薇; 李进杰; 周惠

    2013-01-01

    目的 建立胞壁酰二肽(MDP)诱发烫伤大鼠脓毒症模型.方法 选择SPF级健康雄性SD大鼠50只,体重200~250 g,2~3月龄,采用随机数字表法,将其分为3组:对照组(C组,n=10)、烫伤组(S组,n=10),制备20%总体表面积Ⅲ度烫伤模型;MDP组(n=30),于烫伤后24h时经股静脉注射MDP 5 mg/kg.MDP组于注射MDP后1、6、24h时,S组于烫伤后24h时,C组于20℃水中假烫伤后24h时取腹主动脉血样,分别行血气分析、计数WBC和Plt、检测肝肾功能、肌酸激酶同工酶(CK-MB)活性、血浆TNF-α、干扰素-γ(IFN-γ)、IL-6、IL-10、高迁移率族蛋白-1(HMGB-1)水平;取血样后处死大鼠,取心、肝、肺和肾组织行病理学检查,另取肺组织测定髓过氧化物酶(MPO)活性.另取雄性SD大鼠90只,按照上述方法分组处理后,记录72 h生存情况.结果 与C组比较,S组血浆IL-6、IL-10、IFN-γ和HMGB1水平、WBC计数、血清ALT、AST和BUN水平、肺组织MPO活性升高,生存率降低,MDP组血浆TNF-α、IL-6、IL-10、IFN-γ和HMGB-1水平、血清ALT、AST、总胆红素、BUN、Cr和CK-MB水平、肺组织MPO活性、PaCO2升高,BE负值增加,WBC和Plt计数、pH值、PaO2和生存率降低(P<0.05).与S组比较,MDP组血浆TNF-α、IL-6、IFN-γ和HMGB-1水平,血清ALT、AST、总胆红素、BUN、Cr、CK-MB水平、肺组织MPO活性和PaC02升高,BE负值增加,WBC和Plt计数、pH值、PaO2和生存率降低(P<0.05).S组和MDP组心、肝、肺和肾组织病理学损伤明显,MDP组病理学损伤程度较重.结论 Ⅲ度烫伤后经MDP诱导可致大鼠广泛炎性反应并伴多器官功能损伤,成功建立烫伤后脓毒症模型.%Objective To establish a rat model of sepsis induced by muramyl dipeptide (MDP) after scald burn.Methods Fifty SPF male Sprague-Dawley rats,aged 2-3 months,weighing 200-250 g,were randomly divided into 3 groups:control group (group C,n =10),scald group (group S,n =10) and MDP group (n =30).The rats were subjected

  12. An Alternative Approach to Combination Vaccines: Intradermal Administration of Isolated Components for Control of Anthrax, Botulism, Plague and Staphylococcal Toxic Shock

    Science.gov (United States)

    2008-09-03

    antigens CaF1 and LcrV, previously shown to protect mice against plague [18,19]. The bubonic form of plague results from Yersinia pestis injected into...GP, Adamovicz J, Friedlander AM: Protection against experimental bubonic and pneumonic plague by a recombinant capsular F1-V antigen fusion protein...intradermal administration of isolated components for control of anthrax, botulism, plague and staphylococcal toxic shock Garry L Morefield1, Ralph F

  13. Identification of a novel variant of staphylococcal cassette chromosome mec, type II.5, and Its truncated form by insertion of putative conjugative transposon Tn6012.

    Science.gov (United States)

    Han, Xiao; Ito, Teruyo; Takeuchi, Fumihiko; Ma, Xiao Xue; Takasu, Michihiko; Uehara, Yoshio; Oliveira, Duarte C; de Lencastre, Hermínia; Hiramatsu, Keiichi

    2009-06-01

    We identified two novel staphylococcal cassette chromosome mec (SCCmec) elements in sequence type 8 methicillin-resistant Staphylococcus aureus strains isolated in Japan: type II.5 SCCmec, whose J1 region was highly homologous to that of type I.2 SCCmec of strain PL72 (previously isolated in Poland), and its J1 region variant caused by the deletion/insertion of putative conjugative transposon Tn6012, identified in four S. aureus genomes.

  14. Nationwide German Multicenter Study on Prevalence of Antibiotic Resistance in Staphylococcal Bloodstream Isolates and Comparative In Vitro Activities of Quinupristin-Dalfopristin

    OpenAIRE

    Von Eiff, Christof; Reinert, Ralf René; Kresken, Michael; Brauers, Johannes; Hafner, Dieter; Peters, Georg

    2000-01-01

    Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448 S. aureus isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the ...

  15. Production of Tumor Necrosis Factor Alpha in Human T Lymphocytes by Staphylococcal Enterotoxin B Correlates with Toxin-Induced Proliferation and Is Regulated through Protein Kinase C

    Science.gov (United States)

    1999-12-01

    molecules. Immunol. Rev. 131:43-59. Chatila, T., N. Wood, J. Parsonnet, and R. S. Geha. 1988. Toxic shock syndrome toxin-1 induces inositol phospholipid...Dohlsten, U. Andersson, G. Hedlund, P. Ericsson, J. Hans- son, and H. O. Sjogren . 1990. Production of TNF-alpha and TNF-beta by staphylococcal... syndrome . Immunobiology 189:270-284. 33. Miethke, T., C. Wahl, K. Heeg, B. Echtenacher, P. H. Krammer, and H. Wagner. 1992. T cell-mediated lethal

  16. Novel staphylococcal glycosyltransferases SdgA and SdgB mediate immunogenicity and protection of virulence-associated cell wall proteins.

    Directory of Open Access Journals (Sweden)

    Wouter L W Hazenbos

    Full Text Available Infection of host tissues by Staphylococcus aureus and S. epidermidis requires an unusual family of staphylococcal adhesive proteins that contain long stretches of serine-aspartate dipeptide-repeats (SDR. The prototype member of this family is clumping factor A (ClfA, a key virulence factor that mediates adhesion to host tissues by binding to extracellular matrix proteins such as fibrinogen. However, the biological siginificance of the SDR-domain and its implication for pathogenesis remain poorly understood. Here, we identified two novel bacterial glycosyltransferases, SdgA and SdgB, which modify all SDR-proteins in these two bacterial species. Genetic and biochemical data demonstrated that these two glycosyltransferases directly bind and covalently link N-acetylglucosamine (GlcNAc moieties to the SDR-domain in a step-wise manner, with SdgB appending the sugar residues proximal to the target Ser-Asp repeats, followed by additional modification by SdgA. GlcNAc-modification of SDR-proteins by SdgB creates an immunodominant epitope for highly opsonic human antibodies, which represent up to 1% of total human IgG. Deletion of these glycosyltransferases renders SDR-proteins vulnerable to proteolysis by human neutrophil-derived cathepsin G. Thus, SdgA and SdgB glycosylate staphylococcal SDR-proteins, which protects them against host proteolytic activity, and yet generates major eptopes for the human anti-staphylococcal antibody response, which may represent an ongoing competition between host and pathogen.

  17. Incidence of Salmonella, Listeria monocytogenes, Escherichia coli O157:H7, and Staphylococcal enterotoxin in two types of Mexican fresh cheeses.

    Science.gov (United States)

    Torres-Vitela, M R; Mendoza-Bernardo, M; Castro-Rosas, J; Gomez-Aldapa, C A; Garay-Martinez, L E; Navarro-Hidalgo, V; Villarruel-López, A

    2012-01-01

    Handcrafted fresh cheeses are popular among consumers in Mexico. However, unsafe raw materials and inadequate food safety practices during cheese manufacture and preservation make them a potential public health risk. The incidence of Salmonella, Listeria, Escherichia coli O157:H7, and staphylococcal enterotoxin was analyzed in two types of fresh cheese (panela and adobera) commonly marketed in Mexico. A total of 200 samples, 100 panela and 100 adobera, were acquired from 100 wholesale milk product distributors who supply small retailers in the Guadalajara metropolitan area, Jalisco State, Mexico. Pathogens were identified using culture and immunoassay (miniVidas) methods. The presence of staphylococcal enterotoxin was determined by an immunoassay method. Of the 200 analyzed samples, 92 were positive for at least one of the pathogens. The incidence in the panela samples was 56%: 34% Salmonella, 16% E. coli O157:H7, and 6% L. monocytogenes. In the adobera samples, incidence was 36%: 20% Salmonella, 4% E. coli O157:H7, and 12% L. monocytogenes. Staphylococcal enterotoxin was not detected in any of the 200 samples. Choice of technique had no effect on detection of pathogen incidence, although the immunoassay method identified more Salmonella serotypes than the culture method. Handcrafted panela and adobera fresh cheeses in Mexico frequently contain pathogenic bacteria and therefore pose a public health risk.

  18. Application of a SERS-based lateral flow immunoassay strip for the rapid and sensitive detection of staphylococcal enterotoxin B

    Science.gov (United States)

    Hwang, Joonki; Lee, Sangyeop; Choo, Jaebum

    2016-06-01

    A novel surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFA) biosensor was developed to resolve problems associated with conventional LFA strips (e.g., limits in quantitative analysis and low sensitivity). In our SERS-based biosensor, Raman reporter-labeled hollow gold nanospheres (HGNs) were used as SERS detection probes instead of gold nanoparticles. With the proposed SERS-based LFA strip, the presence of a target antigen can be identified through a colour change in the test zone. Furthermore, highly sensitive quantitative evaluation is possible by measuring SERS signals from the test zone. To verify the feasibility of the SERS-based LFA strip platform, an immunoassay of staphylococcal enterotoxin B (SEB) was performed as a model reaction. The limit of detection (LOD) for SEB, as determined with the SERS-based LFA strip, was estimated to be 0.001 ng mL-1. This value is approximately three orders of magnitude more sensitive than that achieved with the corresponding ELISA-based method. The proposed SERS-based LFA strip sensor shows significant potential for the rapid and sensitive detection of target markers in a simplified manner.A novel surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFA) biosensor was developed to resolve problems associated with conventional LFA strips (e.g., limits in quantitative analysis and low sensitivity). In our SERS-based biosensor, Raman reporter-labeled hollow gold nanospheres (HGNs) were used as SERS detection probes instead of gold nanoparticles. With the proposed SERS-based LFA strip, the presence of a target antigen can be identified through a colour change in the test zone. Furthermore, highly sensitive quantitative evaluation is possible by measuring SERS signals from the test zone. To verify the feasibility of the SERS-based LFA strip platform, an immunoassay of staphylococcal enterotoxin B (SEB) was performed as a model reaction. The limit of detection (LOD) for SEB, as

  19. Effects of burn oil of Chinese medicinal herbs on wound healing in scalded rats%中药烧伤油对大鼠深Ⅱ度烫伤创面愈合的作用

    Institute of Scientific and Technical Information of China (English)

    杨军; 张殿增; 刘晓瑾; 白转丽; 郭敏峰; 王瑞; 彭慧子; 段辉

    2009-01-01

    AIM: To investigate the effects of burn oil of Chinese medicinal herbs on wound healing in scalded rats. METHODS: Three deep-Ⅱ-degree circular skin wounds with a diameter of 2.5 cm were prepared in the back of rats and were treated respectively with burn oil of Chinese medicinal herbs, zinc sulfadiasine cream and matrix ointment 1 time per day. The healing of the wounds was observed. The wound tissue samples were harvested at different times for histopathological examination and the content of hydroxyproline, type Ⅰ/Ⅱ collagen ratio and the fibrocyte eyele of the dermal wound were detected by ELISA assay and flow cytometry. RESULTS : The application of burn oil of Chinese medicinal herbs reduced the swelling and effusion and shortened the healing time of the wound with no wound infection. Pathological examination showed that bum oil of Chinese medicinal herbs alleviated topical inflammation. All layers of derma were well-differentiated. Effect of hydroxyproline and wound type Ⅰ/Ⅲ collagen ratio showed that bum oil of Chinese medicinal herbs promoted the collage hyperplasy of wounds and palliated scars. The observation of the fibrocyte cycle showed that burn oil of Chinese medicinal herbs promoted the dermal fibrocyte proliferation. CONCLUSION: Burn oil of Chinese medicinal herbs promotes the wound healing in scalded rats.%目的:探讨中药烧伤油对大鼠深Ⅱ度烫伤创面愈合的影响.方法:制备深Ⅱ度大鼠背部皮肤圆形创面,直径2.5 cm,共3个,行同体对照实验.分别外用烧伤油,磺胺嘧啶锌软膏和基质软膏,1次/d.观察创面愈合情况,留取不同时相的创面组织,进行组织病理学检查,创面羟脯氨酸含量检测,ELISA法测定创面Ⅰ/Ⅱ型胶原比例和流式细胞仪测定创面真皮成纤维细胞周期.结果:外用烧伤油使创面肿胀和渗出减轻,无感染迹象及创面愈合时间缩短.病理形态学观察发现烧伤油组创面的局部组织炎症

  20. Effects of escharectomy at different time points on myocardial damages in scalded rats during shock stage%烫伤大鼠休克期内不同时间切痂对心肌损伤的影响

    Institute of Scientific and Technical Information of China (English)

    张会堂; 王甲汉

    2001-01-01

    Objective To investigate the effects of escharectomy at different time points on myocardial damages in scalded rats during shock stage. Methods A total of 42 rats were inflicted with 30% TBSA Ⅲ scalding on their backs and then randomly and equally divided into 7 groups: burned control group (C) and escharectomy groups at hour 1, 3, 6, 12, 24 and 36 postburn. Another 6 rats were employed as normal control (N). The serum levels of creatine kinase isoenzymes MB (CKMB), aspartate aminotransferase (AST) were determined and morphological changes in myocardial tissues were observed 72 hours after the burn. Results The serum levels of CKMB and AST were higher in the C group than in other groups, and the levels in escharectomy groups were increased along with the prolongation of scar cutting. Pathologically, the myocardial tissues were severely damaged in C group, and the later the scars were cut, the severe the damages were in the escharectomy groups except the 2 groups with scars cut at hour 1 and 3. Conclusion Escharectomy in shock stage can effectively prevent the damage of the myocardial tissues from postburn injury and the earlier it is performed, the better the result is.%目的 探讨烧伤后休克期内不同时间切痂对心肌损害的影响。方法 以大鼠30%TBSAⅢ度烫伤为模型,动物随机分为正常对照组(N组,n=6),烧伤对照组(C组,n=6),1、3、6、12、24和36 h切痂组(n=6),各切痂组于伤后各时相点切痂,伤后72 h行血清肌酸激酶同工酶MB(CKMB)、天冬氨酸氨基转换酶(AST)检测和心肌组织病理观察。结果 C组血清CKMB、AST浓度显著高于其余各组,心肌组织病理变化程度重,各切痂组血清CKMB、AST浓度随伤后切痂时间延长而逐渐升高,心肌病理改变逐渐加重,而l、3 h切痂组与正常对照组比较改变不明显。结论 休克期切痂能有效防治烧伤后心肌损害,且切痂越早,效果越好。

  1. Effects of Rhodomyrtus tomentosa Leaf Extract on Staphylococcal Adhesion and Invasion in Bovine Udder Epidermal Tissue Model.

    Science.gov (United States)

    Mordmuang, Auemphon; Shankar, Shiv; Chethanond, Usa; Voravuthikunchai, Supayang Piyawan

    2015-10-15

    Bovine mastitis is one of the most important infectious diseases in dairy herds, and staphylococci are the most important etiologic agents of this disease. Antibiotics and chemical agents used in livestock for prevention and cure of the disease can accumulate in milk and give rise to food safety concerns. Rhodomyrtus tomentosa leaf extract was studied as an alternative approach to reduce the bacterial infections. The ethanolic extract of this plant demonstrated antibacterial activity with minimum inhibitory concentration (MIC) values as low as 16-64 μg/mL against staphylococcal isolates. In addition, the extract had an effect on the bacterial cell surface properties by increasing its hydrophobicity in a concentration dependent manner. To further extend the antibacterial efficacy, silver nanoparticles synthesized with the extract, a pure rhodomyrtone, and liposomal encapsulated rhodomyrtone were applied and their inhibitory effects on bacterial adhesion and invasion were determined by ex vivo study in a bovine udder epidermal tissue model. These agents exerted remarkable antibacterial activity against staphylococci and decreased the adhesion of the bacterial cells to the tissues. These results supported that R. tomentosa ethanolic extract could be applied as an alternative agent for bovine udder care in dairy farms.

  2. Prophage-Encoded Staphylococcal Enterotoxin A: Regulation of Production in Staphylococcus aureus Strains Representing Different Sea Regions.

    Science.gov (United States)

    Zeaki, Nikoleta; Susilo, Yusak Budi; Pregiel, Anna; Rådström, Peter; Schelin, Jenny

    2015-12-09

    The present study investigates the nature of the link between the staphylococcal enterotoxin A (SEA) gene and the lifecycle of Siphoviridae bacteriophages, including the origin of strain variation regarding SEA production after prophage induction. Five strains representing three different genetic lines of the sea region were studied under optimal and prophage-induced growth conditions and the Siphoviridae lifecycle was followed through the phage replicative form copies and transcripts of the lysogenic repressor, cro. The role of SOS response on prophage induction was addressed through recA transcription in a recA-disruption mutant. Prophage induction was found to increase the abundance of the phage replicative form, the sea gene copies and transcripts and enhance SEA production. Sequence analysis of the sea regions revealed that observed strain variances were related to strain capacity for prophage induction, rather than sequence differences in the sea region. The impact of SOS response activation on the phage lifecycle was demonstrated by the absence of phage replicative form copies in the recA-disruption mutant after prophage induction. From this study it emerges that all aspects of SEA-producing strain, the Siphoviridae phage and the food environment must be considered when evaluating SEA-related hazards.

  3. Cross-linking staphylococcal enterotoxin A bound to major histocompatibility complex class I is required for TNF-alpha secretion

    Science.gov (United States)

    Wright, A. D.; Chapes, S. K.

    1999-01-01

    The mechanism of how superantigens function to activate cells has been linked to their ability to bind and cross-link the major histocompatibility complex class II (MHCII) molecule. Cells that lack the MHCII molecule also respond to superantigens, however, with much less efficiency. Therefore, the purpose of this study was to confirm that staphylococcal enterotoxin A (SEA) could bind the MHCI molecule and to test the hypothesis that cross-linking SEA bound to MHCII-deficient macrophages would induce a more robust cytokine response than without cross-linking. We used a capture enzyme-linked immunosorbent assay and an immunprecipitation assay to directly demonstrate that MHCI molecules bind SEA. Directly cross-linking MHCI using monoclonal antibodies or cross-linking bound SEA with an anti-SEA antibody or biotinylated SEA with avidin increased TNF-alpha and IL-6 secretion by MHCII(-/-) macrophages. The induction of a vigorous macrophage cytokine response by SEA/anti-SEA cross-linking of MHCI offers a mechanism to explain how MHCI could play an important role in superantigen-mediated pathogenesis. Copyright 1999 Academic Press.

  4. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Energy Technology Data Exchange (ETDEWEB)

    Hill, F.S.; Cantu, A.O.; Lucas, J.N. (Lawrence Livermore National Lab., CA (United States)); Cox, A.B.; Salmon, Y.L. (Air Force Armstrong Lab., Brookes AFB, TX (United States))

    1994-10-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 [mu]g/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) that that produced by PHA (M1<0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture. (author).

  5. Staphylococcal food poisoning case and molecular analysis of toxin genes in Staphylococcus aureus strains isolated from food in Sicily, Italy.

    Science.gov (United States)

    Vitale, Maria; Scatassa, Maria Luisa; Cardamone, Cinzia; Oliveri, Giuseppa; Piraino, Chiara; Alduina, Rosa; Napoli, Concetta

    2015-01-01

    A case of staphylococcal food poisoning was observed in two individuals of the same family after consumption of primosale, a semiripened sheep cheese produced in Sicily. Staphylococcus aureus isolated from the cheese produced enterotoxin C (SEC) and carried both the enterotoxin C (sec) and the toxic shock syndrome toxin (tsst-1) gene. Following this case, an extensive survey was conducted on 971 food samples (raw milk, cheese, meat, and food preparations). S. aureus was detected in 102 of 971 food samples, from all types of food with the exception of ricotta cheese. The tsst-1 gene was present in 42% of the strains, either alone or in combination with other toxin genes. The enterotoxin C gene was the most represented enterotoxin, but it was only found in dairy products. Six S. aureus isolates carried the sea gene alone, two isolates carried both sea and seb, and one isolate carried both sea and sec. A significant percentage (46%) of all isolates carried a toxin gene, creating significant concern that virulent S. aureus can be transmitted through food in Sicily.

  6. New silica nanostructure for the improved delivery of topical antibiotics used in the treatment of staphylococcal cutaneous infections.

    Science.gov (United States)

    Grumezescu, Alexandru Mihai; Ghitulica, Cristina Daniela; Voicu, Georgeta; Huang, Keng-Shiang; Yang, Chih-Hui; Ficai, Anton; Vasile, Bogdan Stefan; Grumezescu, Valentina; Bleotu, Coralia; Chifiriuc, Mariana Carmen

    2014-03-25

    In this paper, we report the synthesis, characterization (FT-IR, XRD, BET, HR-TEM) and bioevaluation of a novel γ-aminobutiric acid/silica (noted GABA-SiO₂ or γ-SiO₂) hybrid nanostructure, for the improved release of topical antibiotics, used in the treatment of Staphylococcus aureus infections. GABA-SiO₂ showed IR bands which were assigned to Si-O-Si (stretch mode). The XRD pattern showed a broad peak in the range of 18-30° (2θ), indicating an amorphous structure. Based on the BET analysis, estimations about surface area (438.14 m²/g) and pore diameters (4.76 nm) were done. TEM observation reveals that the prepared structure presented homogeneity and an average size of particles not exceeding 10nm. The prepared nanostructure has significantly improved the anti-staphylococcal activity of bacitracin and kanamycin sulfate, as demonstrated by the drastic decrease of the minimal inhibitory concentration of the respective antibiotics loaded in the GABA-SiO₂ nanostructure. These results, correlated with the high biocompatibility of this porous structure, are highlighting the possibility of using this carrier for the local delivery of the antimicrobial substances in lower active doses, thus reducing their cytotoxicity and side-effects.

  7. Effects of Rhodomyrtus tomentosa Leaf Extract on Staphylococcal Adhesion and Invasion in Bovine Udder Epidermal Tissue Model

    Directory of Open Access Journals (Sweden)

    Auemphon Mordmuang

    2015-10-01

    Full Text Available Bovine mastitis is one of the most important infectious diseases in dairy herds, and staphylococci are the most important etiologic agents of this disease. Antibiotics and chemical agents used in livestock for prevention and cure of the disease can accumulate in milk and give rise to food safety concerns. Rhodomyrtus tomentosa leaf extract was studied as an alternative approach to reduce the bacterial infections. The ethanolic extract of this plant demonstrated antibacterial activity with minimum inhibitory concentration (MIC values as low as 16–64 μg/mL against staphylococcal isolates. In addition, the extract had an effect on the bacterial cell surface properties by increasing its hydrophobicity in a concentration dependent manner. To further extend the antibacterial efficacy, silver nanoparticles synthesized with the extract, a pure rhodomyrtone, and liposomal encapsulated rhodomyrtone were applied and their inhibitory effects on bacterial adhesion and invasion were determined by ex vivo study in a bovine udder epidermal tissue model. These agents exerted remarkable antibacterial activity against staphylococci and decreased the adhesion of the bacterial cells to the tissues. These results supported that R. tomentosa ethanolic extract could be applied as an alternative agent for bovine udder care in dairy farms.

  8. 多重PCR检测MRSA的SCCmec基因分型%Staphylococcal chromosome cassette typing in MRSA by multiplex PCR

    Institute of Scientific and Technical Information of China (English)

    韩玉涛; 蒋燕群

    2008-01-01

    目的 了解我院MRSA的流行状况.方法 收集2005年1-6月65株社区感染MRSA及60株医院感染MRSA,应用多重PCR对MRSA染色体mec基因盒(Staphylococcal cassette chromosome SCCmec)分型及杀白细胞毒素(PVL)基因检测,应用K-B纸片法进行药敏分析.结果 125株MRSA的mecA基因阳性,其中SCCmecⅡ型1株,SCCmecⅢ型120株,SCCmecⅣ型3株,未分型1株;未发现携带PVL基因的MRSA.携带SCCmecⅡ型、SCCmecⅢ型的菌株均为多重耐药株,而携带SCCmecⅣ型的菌株除对β内酰胺类药物耐药外,对其他类别的抗菌药敏感.结论 本院分离的MRSA以SCCmecⅢ型为主,发现SCCmecⅣ型CA-MRSA,但不携带PVL基因;携带SCCmecⅡ、SCCmecⅢ的临床分离株耐药严重.

  9. Staphylococcal superantigen-specific IgE antibodies: degree of sensitization and association with severity of asthma.

    Science.gov (United States)

    Elabras, José; Mello, Fernanda Carvalho de Queiroz; Lupi, Omar; Bica, Blanca Elena Rios Gomes; Papi, José Angelo de Souza; França, Alfeu Tavares

    2016-01-01

    To determine the presence of staphylococcal superantigen-specific IgE antibodies and degree of IgE-mediated sensitization, as well as whether or not those are associated with the severity of asthma in adult patients. This was a cross-sectional study involving outpatients with asthma under treatment at a tertiary care university hospital in the city of Rio de Janeiro, Brazil. Consecutive patients were divided into two groups according to the severity of asthma based on the Global Initiative for Asthma criteria: mild asthma (MA), comprising patients with mild intermittent or persistent asthma; and moderate or severe asthma (MSA). We determined the serum levels of staphylococcal toxin-specific IgE antibodies, comparing the results and performing a statistical analysis. The study included 142 patients: 72 in the MA group (median age = 46 years; 59 females) and 70 in the MSA group (median age = 56 years; 60 females). In the sample as a whole, 62 patients (43.7%) presented positive results for staphylococcal toxin-specific IgE antibodies: staphylococcal enterotoxin A (SEA), in 29 (20.4%); SEB, in 35 (24.6%); SEC, in 33 (23.2%); and toxic shock syndrome toxin (TSST), in 45 (31.7%). The mean serum levels of IgE antibodies to SEA, SEB, SEC, and TSST were 0.96 U/L, 1.09 U/L, 1.21 U/L, and 1.18 U/L, respectively. There were no statistically significant differences between the two groups in terms of the qualitative or quantitative results. Serum IgE antibodies to SEA, SEB, SEC, and TSST were detected in 43.7% of the patients in our sample. However, neither the qualitative nor quantitative results showed a statistically significant association with the clinical severity of asthma. Determinar a presença de anticorpos IgE específicos para superantígenos estafilocócicos e o grau de sensibilização mediada por esses, assim como se esses estão associados à gravidade da asma em pacientes adultos. Estudo transversal incluindo asmáticos adultos em acompanhamento ambulatorial em

  10. Recombinations in staphylococcal cassette chromosome mec elements compromise the molecular detection of methicillin resistance in Staphylococcus aureus

    KAUST Repository

    Hill-Cawthorne, Grant A.

    2014-06-27

    Clinical laboratories are increasingly using molecular tests for methicillin-resistant Staphylococcus aureus (MRSA) screening. However, primers have to be targeted to a variable chromosomal region, the staphylococcal cassette chromosome mec (SCCmec). We initially screened 726 MRSA isolates from a single UK hospital trust by recombinase polymerase amplification (RPA), a novel, isothermal alternative to PCR. Undetected isolates were further characterised using multilocus sequence, spa typing and whole genome sequencing. 96% of our tested phenotypically MRSA isolates contained one of the six orfX-SCCmec junctions our RPA test and commercially available molecular tests target. However 30 isolates could not be detected. Sequencing of 24 of these isolates demonstrated recombinations within the SCCmec element with novel insertions that interfered with the RPA, preventing identification as MRSA. This result suggests that clinical laboratories cannot rely solely upon molecular assays to reliably detect all methicillin-resistance. The presence of significant recombinations in the SCCmec element, where the majority of assays target their primers, suggests that there will continue to be isolates that escape identification. We caution that dependence on amplification-based molecular assays will continue to result in failure to diagnose a small proportion (?4%) of MRSA isolates, unless the true level of SCCmec natural diversity is determined by whole genome sequencing of a large collection of MRSA isolates. © 2014 Hill-Cawthorne et al.

  11. Recombinations in staphylococcal cassette chromosome mec elements compromise the molecular detection of methicillin resistance in Staphylococcus aureus.

    Directory of Open Access Journals (Sweden)

    Grant A Hill-Cawthorne

    Full Text Available Clinical laboratories are increasingly using molecular tests for methicillin-resistant Staphylococcus aureus (MRSA screening. However, primers have to be targeted to a variable chromosomal region, the staphylococcal cassette chromosome mec (SCCmec. We initially screened 726 MRSA isolates from a single UK hospital trust by recombinase polymerase amplification (RPA, a novel, isothermal alternative to PCR. Undetected isolates were further characterised using multilocus sequence, spa typing and whole genome sequencing. 96% of our tested phenotypically MRSA isolates contained one of the six orfX-SCCmec junctions our RPA test and commercially available molecular tests target. However 30 isolates could not be detected. Sequencing of 24 of these isolates demonstrated recombinations within the SCCmec element with novel insertions that interfered with the RPA, preventing identification as MRSA. This result suggests that clinical laboratories cannot rely solely upon molecular assays to reliably detect all methicillin-resistance. The presence of significant recombinations in the SCCmec element, where the majority of assays target their primers, suggests that there will continue to be isolates that escape identification. We caution that dependence on amplification-based molecular assays will continue to result in failure to diagnose a small proportion (∼4% of MRSA isolates, unless the true level of SCCmec natural diversity is determined by whole genome sequencing of a large collection of MRSA isolates.

  12. Methicillin-Resistant Staphylococcus saprophyticus Isolates Carrying Staphylococcal Cassette Chromosome mec Have Emerged in Urogenital Tract Infections▿

    Science.gov (United States)

    Higashide, Masato; Kuroda, Makoto; Omura, Carlos Takashi Neves; Kumano, Miyuki; Ohkawa, Saburo; Ichimura, Sadahiro; Ohta, Toshiko

    2008-01-01

    Staphylococcus saprophyticus is a uropathogenic bacterium that causes acute uncomplicated urinary tract infections, particularly in female outpatients. We investigated the dissemination and antimicrobial susceptibilities of 101 S. saprophyticus isolates from the genitourinary tracts of patients in Japan. Eight of these isolates were mecA positive and showed β-lactam resistance. Pulsed-field gel electrophoresis showed that only some isolates were isogenic, indicating that the mecA gene was apparently acquired independently by mecA-positive isolates through staphylococcal cassette chromosome mec (SCCmec). Type determination of SCCmec by multiplex PCR showed a nontypeable element in the eight mecA-positive isolates. Sequence analysis of the entire SCCmec element from a prototype S. saprophyticus strain revealed that it was nontypeable with the current SCCmec classification due to the novel composition of the class A mec gene complex (IS431-mecA-mecR1-mecI genes) and the ccrA1/ccrB3 gene complex. Intriguingly, the attachment sites of SCCmec are similar to those of type I SCCmec in S. aureus NCTC 10442. Furthermore, the genes around the mec gene complex are similar to those of type II/III SCCmec in S. aureus, while those around the ccr gene complex are similar to those of SCC15305RM found in S. saprophyticus ATCC 15305. In comparison with known SCCmec elements, this S. saprophyticus SCCmec is a novel type. PMID:18362191

  13. Tudor staphylococcal nuclease drives chemoresistance of non-small cell lung carcinoma cells by regulating S100A11.

    Science.gov (United States)

    Zagryazhskaya, Anna; Surova, Olga; Akbar, Nadeem S; Allavena, Giulia; Gyuraszova, Katarina; Zborovskaya, Irina B; Tchevkina, Elena M; Zhivotovsky, Boris

    2015-05-20

    Lung cancer is the leading cause of cancer-related deaths worldwide. Non-small cell lung cancer (NSCLC), the major lung cancer subtype, is characterized by high resistance to chemotherapy. Here we demonstrate that Tudor staphylococcal nuclease (SND1 or TSN) is overexpressed in NSCLC cell lines and tissues, and is important for maintaining NSCLC chemoresistance. Downregulation of TSN by RNAi in NSCLC cells led to strong potentiation of cell death in response to cisplatin. Silencing of TSN was accompanied by a significant decrease in S100A11 expression at both mRNA and protein level. Downregulation of S100A11 by RNAi resulted in enhanced sensitivity of NSCLC cells to cisplatin, oxaliplatin and 5-fluouracil. AACOCF(3), a phospholipase A(2) (PLA(2)) inhibitor, strongly abrogated chemosensitization upon silencing of S100A11 suggesting that PLA(2) inhibition by S100A11 governs the chemoresistance of NSCLC. Moreover, silencing of S100A11 stimulated mitochondrial superoxide production, which was decreased by AACOCF(3), as well as N-acetyl-L-cysteine, which also mimicked the effect of PLA(2) inhibitor on NSCLC chemosensitization upon S100A11 silencing. Thus, we present the novel TSN-S100A11-PLA(2) axis regulating superoxide-dependent apoptosis, triggered by platinum-based chemotherapeutic agents in NSCLC that may be targeted by innovative cancer therapies.

  14. Evaluation of the in vitro activities of ceftobiprole and comparators in staphylococcal colony or microtitre plate biofilm assays.

    Science.gov (United States)

    Abbanat, Darren; Shang, Wenchi; Amsler, Karen; Santoro, Colleen; Baum, Ellen; Crespo-Carbone, Steven; Lynch, A Simon

    2014-01-01

    The aim of this study was to evaluate the in vitro efficacy of ceftobiprole and comparator antibiotics, either alone or in combination, in staphylococcal MBEC™ (minimum biofilm eradication concentration) and colony biofilm assays at dilutions of the maximum free-drug plasma concentration attained during clinical use (fCmax). Staphylococci tested included meticillin-susceptible and meticillin-resistant Staphylococcus aureus (n=6) and Staphylococcus epidermidis (n=2). Relative to no-drug controls, after 7 days of exposure ceftobiprole concentrations from 1/4 fCmax to fCmax generally decreased CFUs in MBEC or colony biofilms of S. aureus isolates by ca. 1.5log10 to ≥2.5log10. Gentamicin reduced colony biofilm CFUs by ≥1.4log10 at these concentrations with gentamicin-susceptible isolates. Following 7 days of exposure, vancomycin and rifampicin were ineffective as single agents or in combination in the colony model, but yielded CFU decreases from 0 to 5log10 in the MBEC model. Treatment of biofilms with rifampicin for 7 days yielded rifampicin-resistant mutants, and the selection of rifampicin resistance was inhibited by co-treatment with ceftobiprole. Thus, ceftobiprole alone or in combination demonstrated promising activity against biofilms of meticillin-susceptible and -resistant staphylococci at clinically relevant concentrations. In contrast, vancomycin and rifampicin, two agents used clinically for the treatment of biofilm infections, tested separately or together gave inconsistent results and generally had little impact on cell viability.

  15. Development of Cell Lines Stably Expressing Staphylococcal Nuclease Fused to Dengue 2 Virus Capsid Protein for CTVI

    Institute of Scientific and Technical Information of China (English)

    Cheng-Feng QIN; E-De QIN

    2004-01-01

    To explore the potential application of capsid-targeted viral inactivation(CTVI)strategy in prophylactic model against dengue virus(DV)infection,here we fused a Ca2+-dependent nuclease,staphylococcal nuclease(SN),to the capsid protein of dengue 2 virus(D2C)at the carboxyl terminal,and constructed the desired expression plasmid pc/D2C-SN and control plasmids pc/D2C-SN* and pc/D2C.A mammalian cell line BHK-21 was transfected by electroporation with those plasmids and thereafter selected by 5 μg/ml blasticidin.The resistant cell clones were then expanding cultured and screened by RT-PCR and Western Blot assays.The nuclease activity of the expressed fusion protein D2C-SN was analyzed by in vitro DNA digestion assay.It was confirmed cell lines stably expressing D2C-SN and control constructs were obtained.The intracellular expressed fusion protein D2C-SN had ideal nuclease activity and no cytotoxicity on mammalian cells.Those engineered cell lines provided the experimental system for CTVI application in prophylactic model and paved the new road for combating DV infection with CTVI.

  16. A rapid radioimmunoassay using /sup 125/I-labeled staphylococcal protein A for antibody to varicella-zoster virus

    Energy Technology Data Exchange (ETDEWEB)

    Richman, D.D.; Cleveland, P.H.; Oxman, M.N.; Zaia, J.A.

    1981-05-01

    A sensitive radioimmunoassay for serum antibody to varicella-zoster virus is described; it uses 125I-labeled staphylococcal protein A and a specially designed immunofiltration apparatus. The assay accurately distinguishes between individuals who are susceptible and those who are immune to infection with varicella-zoster virus. In addition, it can detect passive antibody in recipients of varicella-zoster immune globulin. This radioimmunoassay also detects the heterologous antibody responses that occasionally occur in patients infected with herpes simplex virus, which also have been detected by other antibody assays. The particular advantages of this assay are the use of noninfectious reagents, the speed of execution (less than 3 hr), the requirement for only small quantities of serum (30 microliters), the objectivity of end-point determination, and the capability of screening large numbers of sera. Consequently, this radioimmunoassay is especially useful for the rapid identification of susceptible individuals, which is essential for the appropriate management of patients and hospital personnel after exposure to varicella.

  17. Staphylococcal α-hemolysin is neurotoxic and causes lysis of brain cells in vivo and in vitro.

    Science.gov (United States)

    Dahlberg, Daniel; Mariussen, Espen; Goverud, Ingeborg Løstegaard; Tønjum, Tone; Mæhlen, Jan; Antal, Ellen-Ann; Hassel, Bjørnar

    2015-05-01

    Formation of a bacterial brain abscess entails loss of brain cells and formation of pus. The mechanisms behind the cell loss are not fully understood. Staphylococcus aureus, a common cause of brain abscesses, produces various exotoxins, including α-hemolysin, which is an important factor in brain abscess formation. α-Hemolysin may cause cytolysis by forming pores in the plasma membrane of various eukaryotic cells. However, whether α-hemolysin causes lysis of brain cells is not known. Nor is it known whether α-hemolysin in the brain causes cell death through pore formation or by acting as a chemoattractant, recruiting leukocytes and causing inflammation. Here we show that α-hemolysin injected into rat brain causes cell damage and edema formation within 30 min. Cell damage was accompanied by an increase in extracellular concentrations of zinc, GABA, glutamate, and other amino acids, indicating plasma membrane damage, but leukocytic infiltration was not seen 0.5-12h after α-hemolysin injection. This was in contrast to injection of S. aureus, which triggered extensive infiltration with neutrophils within 8h. In vitro, α-hemolysin caused concentration-dependent lysis of isolated nerve endings and cultured astrocytes. We conclude that α-hemolysin contributes to the cell death inherent in staphylococcal brain abscess formation as a pore-forming neurotoxin.

  18. Prophage-Encoded Staphylococcal Enterotoxin A: Regulation of Production in Staphylococcus aureus Strains Representing Different Sea Regions

    Directory of Open Access Journals (Sweden)

    Nikoleta Zeaki

    2015-12-01

    Full Text Available The present study investigates the nature of the link between the staphylococcal enterotoxin A (SEA gene and the lifecycle of Siphoviridae bacteriophages, including the origin of strain variation regarding SEA production after prophage induction. Five strains representing three different genetic lines of the sea region were studied under optimal and prophage-induced growth conditions and the Siphoviridae lifecycle was followed through the phage replicative form copies and transcripts of the lysogenic repressor, cro. The role of SOS response on prophage induction was addressed through recA transcription in a recA-disruption mutant. Prophage induction was found to increase the abundance of the phage replicative form, the sea gene copies and transcripts and enhance SEA production. Sequence analysis of the sea regions revealed that observed strain variances were related to strain capacity for prophage induction, rather than sequence differences in the sea region. The impact of SOS response activation on the phage lifecycle was demonstrated by the absence of phage replicative form copies in the recA-disruption mutant after prophage induction. From this study it emerges that all aspects of SEA-producing strain, the Siphoviridae phage and the food environment must be considered when evaluating SEA-related hazards.

  19. Structural elucidation of the NADP(H) phosphatase activity of staphylococcal dual-specific IMPase/NADP(H) phosphatase.

    Science.gov (United States)

    Bhattacharyya, Sudipta; Dutta, Anirudha; Dutta, Debajyoti; Ghosh, Ananta Kumar; Das, Amit Kumar

    2016-02-01

    NADP(H)/NAD(H) homeostasis has long been identified to play a pivotal role in the mitigation of reactive oxygen stress (ROS) in the intracellular milieu and is therefore critical for the progression and pathogenesis of many diseases. NAD(H) kinases and NADP(H) phosphatases are two key players in this pathway. Despite structural evidence demonstrating the existence and mode of action of NAD(H) kinases, the specific annotation and the mode of action of NADP(H) phosphatases remains obscure. Here, structural evidence supporting the alternative role of inositol monophosphatase (IMPase) as an NADP(H) phosphatase is reported. Crystal structures of staphylococcal dual-specific IMPase/NADP(H) phosphatase (SaIMPase-I) in complex with the substrates D-myo-inositol-1-phosphate and NADP(+) have been solved. The structure of the SaIMPase-I-Ca(2+)-NADP(+) ternary complex reveals the catalytic mode of action of NADP(H) phosphatase. Moreover, structures of SaIMPase-I-Ca(2+)-substrate complexes have reinforced the earlier proposal that the length of the active-site-distant helix α4 and its preceding loop are the predisposing factors for the promiscuous substrate specificity of SaIMPase-I. Altogether, the evidence presented suggests that IMPase-family enzymes with a shorter α4 helix could be potential candidates for previously unreported NADP(H) phosphatase activity.

  20. Methicillin-resistant Staphylococcus saprophyticus isolates carrying staphylococcal cassette chromosome mec have emerged in urogenital tract infections.

    Science.gov (United States)

    Higashide, Masato; Kuroda, Makoto; Omura, Carlos Takashi Neves; Kumano, Miyuki; Ohkawa, Saburo; Ichimura, Sadahiro; Ohta, Toshiko

    2008-06-01

    Staphylococcus saprophyticus is a uropathogenic bacterium that causes acute uncomplicated urinary tract infections, particularly in female outpatients. We investigated the dissemination and antimicrobial susceptibilities of 101 S. saprophyticus isolates from the genitourinary tracts of patients in Japan. Eight of these isolates were mecA positive and showed beta-lactam resistance. Pulsed-field gel electrophoresis showed that only some isolates were isogenic, indicating that the mecA gene was apparently acquired independently by mecA-positive isolates through staphylococcal cassette chromosome mec (SCCmec). Type determination of SCCmec by multiplex PCR showed a nontypeable element in the eight mecA-positive isolates. Sequence analysis of the entire SCCmec element from a prototype S. saprophyticus strain revealed that it was nontypeable with the current SCCmec classification due to the novel composition of the class A mec gene complex (IS431-mecA-mecR1-mecI genes) and the ccrA1/ccrB3 gene complex. Intriguingly, the attachment sites of SCCmec are similar to those of type I SCCmec in S. aureus NCTC 10442. Furthermore, the genes around the mec gene complex are similar to those of type II/III SCCmec in S. aureus, while those around the ccr gene complex are similar to those of SCC15305RM found in S. saprophyticus ATCC 15305. In comparison with known SCCmec elements, this S. saprophyticus SCCmec is a novel type.

  1. Positive Regulation of Staphylococcal Enterotoxin H by Rot (Repressor of Toxin) Protein and Its Importance in Clonal Complex 81 Subtype 1 Lineage-Related Food Poisoning.

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    Sato'o, Yusuke; Hisatsune, Junzo; Nagasako, Yuria; Ono, Hisaya K; Omoe, Katsuhiko; Sugai, Motoyuki

    2015-11-01

    We previously demonstrated the clonal complex 81 (CC81) subtype 1 lineage is the major staphylococcal food poisoning (SFP)-associated lineage in Japan (Y. Sato'o et al., J Clin Microbiol 52:2637-2640, 2014, http://dx.doi.org/10.1128/JCM.00661-14). Strains of this lineage produce staphylococcal enterotoxin H (SEH) in addition to SEA. However, an evaluation of the risk for the recently reported SEH has not been sufficiently conducted. We first searched for staphylococcal enterotoxin (SE) genes and SE proteins in milk samples that caused a large SFP outbreak in Japan. Only SEA and SEH were detected, while there were several SE genes detected in the samples. We next designed an experimental model using a meat product to assess the productivity of SEs and found that only SEA and SEH were detectably produced in situ. Therefore, we investigated the regulation of SEH production using a CC81 subtype 1 isolate. Through mutant analysis of global regulators, we found the repressor of toxin (Rot) functioned oppositely as a stimulator of SEH production. SEA production was not affected by Rot. seh mRNA expression correlated with rot both in media and on the meat product, and the Rot protein was shown to directly bind to the seh promoter. The seh promoter sequence was predicted to form a loop structure and to hide the RNA polymerase binding sequences. We propose Rot binds to the promoter sequence of seh and unfolds the secondary structure that may lead the RNA polymerase to bind the promoter, and then seh mRNA transcription begins. This alternative Rot regulation for SEH may contribute to sufficient toxin production by the CC81 subtype 1 lineage in foods to induce SFP.

  2. 葡萄球菌肠毒素C2的研究进展%Advances in research of staphylococcal enterotoxin C2

    Institute of Scientific and Technical Information of China (English)

    谷丽维; 朱庆; 王君; 江华; 李建春

    2011-01-01

    葡萄球菌肠毒素(staphylococcal enterotoxin,SE)是一种外源性超抗原,仅需微量即能高效刺激T淋巴细胞增殖,促使其产生细胞毒作用,并释放大量的细胞因子,引发机体自身免疫应答.大量研究表明,SE是一种很有前途的新型肿瘤免疫治疗制剂,目前国内已将SEC2应用于肿瘤患者的治疗及对肿瘤患者在进行放化疗时防止白细胞降低.该文综述了SEC2在生物学活性、结构特性、抑制肿瘤细胞生长作用等方面的研究进展.%Staphylococcal enterotoxin ( SE ) is an exogenous superantigen, which can efficiently stimulate proliferation of T lymphocytes, induce cell-mediated cytotoxicity and cytokine production, and improve the body immunity. Various studies demonstrate that SE is a new promising agent for immunotherapy. It has been recently used as a therapy for tumor patients to prevent the decrease of leucocytes during chemotherapy. In this paper, the research progress in staphylococcal enterotoxin C2 ( SEC2 ) in terms of biological activities, structural characteristics and its effect on promoting lymphocyte proliferation and inhibiting tumor cell growth is reviewed.

  3. Tabebuia avellanedae naphthoquinones: activity against methicillin-resistant staphylococcal strains, cytotoxic activity and in vivo dermal irritability analysis

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    Giambiagi-deMarval Marcia

    2006-03-01

    Full Text Available Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA and coagulase-negative staphylococcus infections are a worldwide concern. Currently, these isolates have also shown resistance to vancomycin, the last therapy used in these cases. It has been observed that quinones and other related compounds exhibit antibacterial activity. This study evaluated the antibacterial activity, toxicity and in vivo dermal irritability of lapachol extracted from Tabebuia avellanedae and derivatives against methicillin-resistant staphylococcal isolates. In addition, its mechanism of action was also analyzed. Methods The compounds β-lapachone, 3-hydroxy β N lapachone and α-lapachone were tested to determine the MIC values against methicillin-resistant S. aureus, S. epidermidis and S. haemolyticus strains, being the two last ones hetero-resistant to vancomycin. Experiments of protein synthesis analysis to investigate the naphthoquinones action were assessed. In vitro toxicity to eukaryotic BSC-40 African Green Monkey Kidney cell cultures and in vivo primary dermal irritability in healthy rabbits were also performed. Results The compounds tested showed antibacterial activity (MICs of 8, 4/8 and 64/128 μg/mL to β-lapachone, 3-hydroxy β N lapachone and α-lapachone, respectively, but no bactericidal activity was observed (MBC > 512 μg/mL for all compounds. Although it has been observed toxic effect in eukaryotic cells, the compounds were shown to be atoxic when applied as topic preparations in healthy rabbits. No inhibition of proteins synthesis was observed. Conclusion Our results suggest that quinones could be used in topic preparations against wound infections caused by staphylococci, after major investigation of the pharmacological properties of the compounds. Studies about the use of these compounds on tumoral cells could be carried on, due to their effect in eukaryotic cells metabolism.

  4. In vitro and in vivo comparison of the anti-staphylococcal efficacy of generic products and the innovator of oxacillin

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    Agudelo Maria

    2010-06-01

    Full Text Available Abstract Background Oxacillin continues to be an important agent in the treatment of staphylococcal infections; many generic products are available and the only requirement for their approval is demonstration of pharmaceutical equivalence. We tested the assumption that pharmaceutical equivalence predicts therapeutic equivalence by comparing 11 generics with the innovator product in terms of concentration of the active pharmaceutical ingredient (API, minimal inhibitory (MIC and bactericidal concentrations (MBC, and antibacterial efficacy in the neutropenic mouse thigh infection model. Methods The API in each product was measured by a validated microbiological assay and compared by slope (potency and intercept (concentration analysis of linear regressions. MIC and MBC were determined by broth microdilution according to Clinical and Laboratory Standard Institute (CLSI guidelines. For in vivo efficacy, neutropenic ICR mice were inoculated with a clinical strain of Staphylococcus aureus. The animals had 4.14 ± 0.18 log10 CFU/thigh when treatment started. Groups of 10 mice per product received a total dose ranging from 2.93 to 750 mg/kg per day administered q1h. Sigmoidal dose-response curves were generated by nonlinear regression fitted to Hill equation to compute maximum effect (Emax, slope (N, and the effective dose reaching 50% of the Emax (ED50. Based on these results, bacteriostatic dose (BD and dose needed to kill the first log of bacteria (1LKD were also determined. Results 4 generic products failed pharmaceutical equivalence due to significant differences in potency; however, all products were undistinguishable from the innovator in terms of MIC and MBC. Independently of their status with respect to pharmaceutical equivalence or in vitro activity, all generics failed therapeutic equivalence in vivo, displaying significantly lower Emax and requiring greater BD and 1LKD, or fitting to a non-sigmoidal model. Conclusions Pharmaceutical or in

  5. Diversity of staphylococcal cassette chromosome mec structures in coagulase-negative staphylococci and relationship to drug resistance.

    Science.gov (United States)

    Garza-González, Elvira; López, Daniel; Pezina, Cesar; Muruet, Walter; Bocanegra-García, Virgilio; Muñoz, Ivan; Ramírez, Camilo; Llaca-Díaz, Jorge M

    2010-03-01

    The objective of this study was to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) elements in meticillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from a tertiary-care hospital in Mexico and to examine the relationship to drug resistance. Fifty selected MR-CoNS isolates collected from catheters (n=15), blood (n=15), bone (n=9), bronchial lavage (n=2) and urine (n=2) and one isolate each from an abscess, cerebrospinal fluid, eye, pleural effusion, synovial fluid, tracheal aspirate and wound secretion were examined. Susceptibility testing was performed by the broth microdilution method. SCCmec types were determined by multiplex PCR and PFGE was carried out as described previously for Staphylococcus aureus. Among the MR-CoNS strains studied, the most frequently isolated species were Staphylococcus epidermidis (n=26) and Staphylococcus haemolyticus (n=13). Staphylococcus cohnii (n=5), Staphylococcus hominis (n=3), Staphylococcus sciuri (n=1), Staphylococcus pasteuri (n=1) and the recently described species Staphylococcus pettenkoferi (n=1) were also identified. The most frequent MR-CoNS genotype identified was SCCmec type IVa in S. epidermidis isolates, which also showed a high diversity in their PFGE patterns. A clone was found that amplified both SCCmec III and V elements in five isolates examined. The single MR S. pettenkoferi isolate harboured SCCmec type IVd and the single MR S. pasteuri isolate harboured SCCmec type I. The carriage of SCCmec type III was associated with resistance or intermediate resistance to meropenem (P IVa and the high genetic diversity among MR-CoNS strains. As far as is known, this is the first report describing the newly identified S. pettenkoferi possessing SCCmec IVd and S. pasteuri harbouring SCCmec type I. MR-CoNS harbouring SCCmec type III were found to be more resistant to meropenem.

  6. Staphylococcal Ecb protein and host complement regulator factor H enhance functions of each other in bacterial immune evasion.

    Science.gov (United States)

    Amdahl, Hanne; Jongerius, Ilse; Meri, Taru; Pasanen, Tanja; Hyvärinen, Satu; Haapasalo, Karita; van Strijp, Jos A; Rooijakkers, Suzan H; Jokiranta, T Sakari

    2013-08-15

    Staphylococcus aureus is a major human pathogen causing more than a tenth of all septicemia cases and often superficial and deep infections in various tissues. One of the immune evasion strategies of S. aureus is to secrete proteins that bind to the central complement opsonin C3b. One of these, extracellular complement binding protein (Ecb), is known to interfere directly with functions of C3b. Because C3b is also the target of the physiological plasma complement regulator, factor H (FH), we studied the effect of Ecb on the complement regulatory functions of FH. We show that Ecb enhances acquisition of FH from serum onto staphylococcal surfaces. Ecb and FH enhance mutual binding to C3b and also the function of each other in downregulating complement activation. Both Ecb and the C-terminal domains 19-20 of FH bind to the C3d part of C3b. We show that the mutual enhancing effect of Ecb and FH on binding to C3b depends on binding of the FH domain 19 to the C3d part of C3b next to the binding site of Ecb on C3d. Our results show that Ecb, FH, and C3b form a tripartite complex. Upon exposure of serum-sensitive Haemophilus influenzae to human serum, Ecb protected the bacteria, and this effect was enhanced by the addition of the C-terminal domains 19-20 of FH. This finding indicates that the tripartite complex formation could give additional protection to bacteria and that S. aureus is thereby able to use host FH and bacterial Ecb in a concerted action to eliminate C3b at the site of infection.

  7. T cell activation and retargeting using staphylococcal enterotoxin B and bispecific antibody: an effective in vivo antitumor strategy.

    Science.gov (United States)

    Porter, L E; Nelson, H; Ethem Gecim, I; Rice, D C; Thibault, C; Chapoval, A I

    1997-01-01

    The aim of this work was to test for cure and immunity in a micrometastatic tumor model using in vivo T cell activation with staphylococcal enterotoxin B (SEB) and retargeting with antitumor x anti-CD3 F(ab')2 bispecific antibodies (bsAb). All studies were performed in C3H/HeN mice using syngeneic tumor cell lines. For survival studies, mice were injected intravenously on day 0 with CL62 (a p97-transfected clone of the K1735 murine melanoma tumor). Day-3 treatments included saline (control), SEB (50 gamma g intraperitoneal) with or without bsAb (5 micrograms i.v.). Cured mice, surviving beyond 60 days, were rechallenged with subcutaneous CL62, K1735, or a nonmelanoma control, AG104. SEB activation studies were performed with pulmonary tumor-infiltrating lymphocytes isolated from 10-day established CL62 tumors. Maximal tumor-infiltrating lymphocyte cytotoxicity was demonstrated 24 h following SEB injection, therefore bsAb treatments were administered 24 h after SEB. When survival was examined at 60 days, there were significantly more survivors in the group receiving SEB plus bsAb (70%) compared to the group receiving SEB alone (30%), and the controls (0%) (P = 0.02 and P < 0.01, respectively). Mice cured of CL62 using SEB alone or with bsAb demonstrated equal immunity to CL62, however, mice treated with SEB plus bsAb were more often immune to the p97-parental cell line, K1735(P = 0.001). Ag104 consistently grew in all mice. Results of these studies demonstrate that SEB plus bsAb can be effective, not only in curing tumors but also in providing protective immunity against targeted and non-targeted tumor antigens.

  8. Staphylococcus aureus Nasal Colonization Differs among Pig Lineages and Is Associated with the Presence of Other Staphylococcal Species

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    Koen M. Verstappen

    2017-06-01

    Full Text Available Staphylococcus aureus is a common colonizer in pigs, with methicillin-resistant S. aureus (MRSA in particular being a potential health risk to humans. To reduce the exposure to humans, the colonization in pigs should be reduced. The aim of this study was to quantitatively compare the susceptibility of pig lineages for S. aureus colonization, and if the absence of S. aureus could be associated with the presence or absence of other staphylococcal species. Nasal samples (n = 129 were obtained from seven different pig lineages in the Netherlands, France, and Germany. S. aureus and other staphylococci were enumerated from these samples by real-time (RT-PCR and culture. Associations were explored between the presence of S. aureus and other staphylococci. S. aureus was detected by RT-PCR on all farms and in samples from pigs of all lineages. Twenty-five percent of the pigs from lineage F (from two farms were colonized with S. aureus, while in all other lineages it was more than 50% (p < 0.01. Moreover, in S. aureus-positive samples from pigs of lineage F smaller amounts of S. aureus were found than in other lineages. Staphylococcus sciuri, Staphylococcus cohnii, and Staphylococcus saprophyticus were usually not found in combination with S. aureus in these samples. In conclusion: (i pigs from different genetic lineages have different susceptibilities for colonization with S. aureus. These pigs might contain a genetic factor influencing nasal colonization. (ii Colonization of S. aureus is also associated with the absence of S. sciuri, S. cohnii, or S. saprophyticus. (iii The farm environment seems to influence the presence of S. aureus in pigs.

  9. Modulation of transforming growth factor‑β signaling transducers in colon adenocarcinoma cells induced by staphylococcal enterotoxin B.

    Science.gov (United States)

    Akbari, Abolfazl; Mobini, Gholam Reza; Maghsoudi, Reza; Akhtari, Javad; Faghihloo, Ebrahim; Farahnejad, Zohreh

    2016-01-01

    Colorectal cancer (CRC) is a notable cause of cancer‑associated mortality worldwide, making it a pertinent topic for the study of cancer and its treatment. Staphylococcal enterotoxin B (SEB), an enterotoxin produced by Staphylococcus aureus, has been demonstrated to exert anticancer and antimetastatic effects due to its ability to modify cell immunity and cellular signaling pathways. In the current study, SEB was investigated, including whether it exerts its growth inhibitory effects on colon adenocarcinoma cells. This may occur through the manipulation of a key tumor growth factor, termed transforming growth factor‑β (TGF‑β), and its signaling pathway transducer, Smad2/3. The human colon adenocarcinoma HCT116 cell line was treated with different concentrations of SEB, and cell number was measured using MTT assay at different treatment times. Smad2/3 RNA expression level was analyzed in untreated or SEB‑treated cells using quantitative polymerase chain reaction, which indicated significant differences between cell viability and Smad2/3 expression levels. SEB effectively downregulated Smad2/3 expression in the HCT116 cells at concentrations of 1 and 2 µg/ml (P=0.0021 and P=0.0017, respectively). SEB concentrations that were effective at inhibiting Smad2/3 expression were correlated with those able to inhibit the proliferation of the cancer cells. SEB inhibited Smad2/3 expression at the mRNA level in a concentration‑ and time‑dependent manner. The present study thus proposed SEB as an agent able to significantly reduce Smad2/3 expression in colon cancer cells, provoking moderate TGF‑β growth signaling and the reduction of tumor cell proliferation.

  10. Rapid Emergence and Evolution of Staphylococcus aureus Clones Harboring fusC-Containing Staphylococcal Cassette Chromosome Elements.

    Science.gov (United States)

    Baines, Sarah L; Howden, Benjamin P; Heffernan, Helen; Stinear, Timothy P; Carter, Glen P; Seemann, Torsten; Kwong, Jason C; Ritchie, Stephen R; Williamson, Deborah A

    2016-04-01

    The prevalence of fusidic acid (FA) resistance amongStaphylococcus aureusstrains in New Zealand (NZ) is among the highest reported globally, with a recent study describing a resistance rate of approximately 28%. Three FA-resistantS. aureusclones (ST5 MRSA, ST1 MSSA, and ST1 MRSA) have emerged over the past decade and now predominate in NZ, and in all three clones FA resistance is mediated by thefusCgene. In particular, ST5 MRSA has rapidly become the dominant MRSA clone in NZ, although the origin of FA-resistant ST5 MRSA has not been explored, and the genetic context offusCin FA-resistant NZ isolates is unknown. To better understand the rapid emergence of FA-resistantS. aureus, we used population-based comparative genomics to characterize a collection of FA-resistant and FA-susceptible isolates from NZ. FA-resistant NZ ST5 MRSA displayed minimal genetic diversity and represented a phylogenetically distinct clade within a global population model of clonal complex 5 (CC5)S. aureus In all lineages,fusCwas invariably located within staphylococcal cassette chromosome (SCC) elements, suggesting that SCC-mediated horizontal transfer is the primary mechanism offusCdissemination. The genotypic association offusCwithmecAhas important implications for the emergence of MRSA clones in populations with high usage of fusidic acid. In addition, we found thatfusCwas colocated with a recently described virulence factor (tirS) in dominant NZS. aureusclones, suggesting a fitness advantage. This study points to the likely molecular mechanisms responsible for the successful emergence and spread of FA-resistantS. aureus.

  11. Staphylococcus aureus Nasal Colonization Differs among Pig Lineages and Is Associated with the Presence of Other Staphylococcal Species.

    Science.gov (United States)

    Verstappen, Koen M; Willems, Eveline; Fluit, Ad C; Duim, Birgitta; Martens, Marc; Wagenaar, Jaap A

    2017-01-01

    Staphylococcus aureus is a common colonizer in pigs, with methicillin-resistant S. aureus (MRSA) in particular being a potential health risk to humans. To reduce the exposure to humans, the colonization in pigs should be reduced. The aim of this study was to quantitatively compare the susceptibility of pig lineages for S. aureus colonization, and if the absence of S. aureus could be associated with the presence or absence of other staphylococcal species. Nasal samples (n = 129) were obtained from seven different pig lineages in the Netherlands, France, and Germany. S. aureus and other staphylococci were enumerated from these samples by real-time (RT)-PCR and culture. Associations were explored between the presence of S. aureus and other staphylococci. S. aureus was detected by RT-PCR on all farms and in samples from pigs of all lineages. Twenty-five percent of the pigs from lineage F (from two farms) were colonized with S. aureus, while in all other lineages it was more than 50% (p S. aureus-positive samples from pigs of lineage F smaller amounts of S. aureus were found than in other lineages. Staphylococcus sciuri, Staphylococcus cohnii, and Staphylococcus saprophyticus were usually not found in combination with S. aureus in these samples. (i) pigs from different genetic lineages have different susceptibilities for colonization with S. aureus. These pigs might contain a genetic factor influencing nasal colonization. (ii) Colonization of S. aureus is also associated with the absence of S. sciuri, S. cohnii, or S. saprophyticus. (iii) The farm environment seems to influence the presence of S. aureus in pigs.

  12. Staphylococcal pathogenicity island DNA packaging system involving cos-site packaging and phage-encoded HNH endonucleases.

    Science.gov (United States)

    Quiles-Puchalt, Nuria; Carpena, Nuria; Alonso, Juan C; Novick, Richard P; Marina, Alberto; Penadés, José R

    2014-04-22

    Staphylococcal pathogenicity islands (SaPIs) are the prototypical members of a widespread family of chromosomally located mobile genetic elements that contribute substantially to intra- and interspecies gene transfer, host adaptation, and virulence. The key feature of their mobility is the induction of SaPI excision and replication by certain helper phages and their efficient encapsidation into phage-like infectious particles. Most SaPIs use the headful packaging mechanism and encode small terminase subunit (TerS) homologs that recognize the SaPI-specific pac site and determine SaPI packaging specificity. Several of the known SaPIs do not encode a recognizable TerS homolog but are nevertheless packaged efficiently by helper phages and transferred at high frequencies. In this report, we have characterized one of the non-terS-coding SaPIs, SaPIbov5, and found that it uses two different, undescribed packaging strategies. SaPIbov5 is packaged in full-sized phage-like particles either by typical pac-type helper phages, or by cos-type phages--i.e., it has both pac and cos sites--a configuration that has not hitherto been described for any mobile element, phages included--and uses the two different phage-coded TerSs. To our knowledge, this is the first example of SaPI packaging by a cos phage, and in this, it resembles the P4 plasmid of Escherichia coli. Cos-site packaging in Staphylococcus aureus is additionally unique in that it requires the HNH nuclease, carried only by cos phages, in addition to the large terminase subunit, for cos-site cleavage and melting.

  13. Application of r-PFE hyperimmune sera for concurrent detection of Bacillus anthracis, Yersinia pestis and staphylococcal enterotoxin B.

    Science.gov (United States)

    Balakrishna, K; Tuteja, U; Murali, H S; Batra, H V

    2014-06-01

    To evaluate the potential of an intergeneric multidomain recombinant chimeric protein for the simultaneous detection of Bacillus anthracis, Yersinia pestis and Staphylococcal enterotoxin B. Truncated portions of protective antigen (pag) of B. anthracis, fraction 1 capsular antigen (F1) of Y. pestis and enterotoxin B (entB) of Staphylococcus aureus were PCR amplified and linked each other using ligation-dependent cloning. The fusion gene was codon-optimized for expression in Escherichia coli and encoded a 55 kDa recombinant PFE protein (rPFE). Hyperimmune antiserum raised against rPFE specifically reacted individually with the native PA of B. anthracis, F1 antigen of Y. pestis and SEB of S. aureus on Western blot analysis as well as in enzyme-linked immunosorbent assay (ELISA). For simultaneous detection of these three antigens from culture supernatants, common media consisting of BHI broth supplemented with 0·2% xylose were used. To assess the detection capability, a known number of these organisms (10(8) -10(2) CFU ml(-1)) were experimentally spiked on to the meat and blood samples, the polyclonal antibodies were again clearly able to identify all three target proteins up to a dilution of 10(5) CFU ml(-1). This recombinant chimeric protein-based immunodetection approach may eventually provide advantages over PCR formats during onsite investigations of biological emergencies or even during routine testing by laboratories. The trivalent recombinant PFE protein could be a novel intervention for possible diagnosis/detection of potential biological agents simultaneously in environmental and clinical samples to reduce the responding time and minimize the impact of the bioattack. © 2014 The Society for Applied Microbiology.

  14. Increasing the frequency of hand washing by healthcare workers does not lead to commensurate reductions in staphylococcal infection in a hospital ward

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    Kerr Kevin G

    2008-09-01

    Full Text Available Abstract Background Hand hygiene is generally considered to be the most important measure that can be applied to prevent the spread of healthcare-associated infection (HAI. Continuous emphasis on this intervention has lead to the widespread opinion that HAI rates can be greatly reduced by increased hand hygiene compliance alone. However, this assumes that the effectiveness of hand hygiene is not constrained by other factors and that improved compliance in excess of a given level, in itself, will result in a commensurate reduction in the incidence of HAI. However, several researchers have found the law of diminishing returns to apply to hand hygiene, with the greatest benefits occurring in the first 20% or so of compliance, and others have demonstrated that poor cohorting of nursing staff profoundly influences the effectiveness of hand hygiene measures. Collectively, these findings raise intriguing questions about the extent to which increasing compliance alone can further reduce rates of HAI. Methods In order to investigate these issues further, we constructed a deterministic Ross-Macdonald model and applied it to a hypothetical general medical ward. In this model the transmission of staphylococcal infection was assumed to occur after contact with the transiently colonized hands of HCWs, who, in turn, acquire contamination only by touching colonized patients. The aim of the study was to evaluate the impact of imperfect hand cleansing on the transmission of staphylococcal infection and to identify, whether there is a limit, above which further hand hygiene compliance is unlikely to be of benefit. Results The model demonstrated that if transmission is solely via the hands of HCWs, it should, under most circumstances, be possible to prevent outbreaks of staphylococcal infection from occurring at a hand cleansing frequencies Conclusion Although our study confirmed hand hygiene to be an effective control measure, it demonstrated that the law of

  15. 伏立诺他对50%TBSAⅢ度烫伤大鼠脏器功能及血流量的影响%Effects of intraperitoneal injection of suberoylanilide hydroxamic acid on organ function and visceral blood flow in rats with full-thickness scald injury involving 50% total body surface area

    Institute of Scientific and Technical Information of China (English)

    郑金光; 胡森; 李芳菲; 张聪; 高明娟; 王晓娜; 姚龙强; 白晓东

    2014-01-01

    目的::探讨组蛋白去乙酰化酶抑制剂伏立诺他(SAHA)对50%TBSA Ⅲ度烫伤大鼠器官功能及脏器血流量的影响。方法:雄性 SD大鼠48只,体重240~260 g,随机分为3组:①单烫组,于100℃水浴中背部浸泡15 s、腹部浸泡8 s,烫伤后,立即腹腔内注射0.25 ml 生理盐水;②SAHA组,烫伤后立即腹腔内注射0.25 ml SAHA注射液(7.5 mg/kg,溶于4 ml 生理盐水中);③假烫组,37℃水浴浸泡后腹腔内注射0.25 ml 生理盐水。于伤后3 h和6 h采用多普勒血流仪测定肝、肾及小肠黏膜血流量,取腹主动脉血检测血清丙氨酸转氨酶(ALT)含量、肌酐(Cr)含量、肌酸激酶同工酶(CK-MB)活性、神经特异性烯醇化酶(NSE)含量和二胺氧化酶(DAO)活性。结果:伤后3 h,SAHA组肝、肾和小肠黏膜血流量显著高于单烫组(P均<0.05);SAHA组6 h小肠黏膜血流量也显著高于单烫组(P<0.05)。SAHA 组伤后3 h Cr、NSE 含量和 DAO 活性显著低于单烫组(P 均<0.05),伤后6 h CK-MB、NSE 含量和 DAO 活性均显著低于单烫组(P 均<0.05)。结论:SAHA 能改善50%TBSAⅢ度烫伤大鼠器官功能指标以及腹腔脏器血流量,保护重要脏器功能。%Objective:To investigate the protective effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA)against injury to organ function and visceral blood flow in rats with 50%total body surface area (TBSA)full-thick-ness scald injury.Methods:Forty-eight male SD rats,weighting 240-260 g,were randomly divided into three groups. In scald group,rats were subj ected to scald inj ury by immersing the back of the trunk for 1 5 seconds and the abdomen for 8 seconds in 100°C water,resulting in full-thickness injury involving 50% TBSA,then they received 0.25 ml of normal saline (NS) intraperitoneally. In SAHA group,rats were subj ected to same extent of inj ury,but they received an intraperitoneal inj ection of 0.25 ml of SAHA (7.5 mg/kg,dissolved in 4 ml of NS). In

  16. Affinity purification of staphylococcal toxic shock syndrome toxin 1 and its pathologic effects in rabbits.

    Science.gov (United States)

    Reeves, M W; Arko, R J; Chandler, F W; Bridges, N B

    1986-01-01

    the heart, lungs, trachea, and thymus. The systemic pathology produced by TSST-1 was strikingly similar to that seen in humans who had died of toxic shock syndrome and in rabbits with subcutaneous chamber inoculated with toxic shock case strains of Staphylococcus aureus. Rabbits that were not killed by the toxin suffered a very rapid and severe leukopenia followed by leukocytosis with a left shift. Lymphopenia was also noted as was a mild but persistent anemia. With the exception of the early leukopenia, very similar hematologic findings have been noted in humans with toxic shock syndrome. Images PMID:3943896

  17. Activity of daptomycin- and vancomycin-loaded poly-epsilon-caprolactone microparticles against mature staphylococcal biofilms

    Directory of Open Access Journals (Sweden)

    Santos Ferreira I

    2015-07-01

    Full Text Available Inês Santos Ferreira,1 Ana F Bettencourt,1 Lídia MD Gonçalves,1 Stefanie Kasper,2 Bertrand Bétrisey,3 Judith Kikhney,2 Annette Moter,2 Andrej Trampuz,4 António J Almeida1 1Research Institute for Medicines (iMed.ULisboa, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal; 2Biofilmcenter, German Heart Institute Berlin, Berlin, Germany; 3Infectious Diseases Service, Department of Medicine, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; 4Center for Musculoskeletal Surgery, Charité – University Medicine Berlin, Berlin, Germany Abstract: The aim of the present study was to develop novel daptomycin-loaded poly-epsilon-caprolactone (PCL microparticles with enhanced antibiofilm activity against mature biofilms of clinically relevant bacteria, methicillin-resistant Staphylococcus aureus (MRSA and polysaccharide intercellular adhesin-positive Staphylococcus epidermidis. Daptomycin was encapsulated into PCL microparticles by a double emulsion-solvent evaporation method. For comparison purposes, formulations containing vancomycin were also prepared. Particle morphology, size distribution, encapsulation efficiency, surface charge, thermal behavior, and in vitro release were assessed. All formulations exhibited a spherical morphology, micro­meter size, and negative surface charge. From a very early time stage, the released concentrations of daptomycin and vancomycin were higher than the minimal inhibitory concentration and continued so up to 72 hours. Daptomycin presented a sustained release profile with increasing concentrations of the drug being released up to 72 hours, whereas the release of vancomycin stabilized at 24 hours. The antibacterial activity of the microparticles was assessed by isothermal microcalorimetry against planktonic and sessile MRSA and S. epidermidis. Regarding planktonic bacteria, daptomycin-loaded PCL microparticles presented the highest antibacterial activity against both strains. Isothermal

  18. Tudor Staphylococcal Nuclease (Tudor-SN) Participates in Small Ribonucleoprotein (snRNP) Assembly via Interacting with Symmetrically Dimethylated Sm Proteins*

    Science.gov (United States)

    Gao, Xingjie; Zhao, Xiujuan; Zhu, Yu; He, Jinyan; Shao, Jie; Su, Chao; Zhang, Yi; Zhang, Wei; Saarikettu, Juha; Silvennoinen, Olli; Yao, Zhi; Yang, Jie

    2012-01-01

    Human Tudor staphylococcal nuclease (Tudor-SN) is composed of four tandem repeats of staphylococcal nuclease (SN)-like domains, followed by a tudor and SN-like domain (TSN) consisting of a central tudor flanked by two partial SN-like sequences. The crystal structure of the tudor domain displays a conserved aromatic cage, which is predicted to hook methyl groups. Here, we demonstrated that the TSN domain of Tudor-SN binds to symmetrically dimethylarginine (sDMA)-modified SmB/B′ and SmD1/D3 core proteins of the spliceosome. We demonstrated that this interaction ability is reduced by the methyltransferase inhibitor 5-deoxy-5-(methylthio)adenosine. Mutagenesis experiments indicated that the conserved amino acids (Phe-715, Tyr-721, Tyr-738, and Tyr-741) in the methyl-binding cage of the TSN domain are required for Tudor-SN-SmB interaction. Furthermore, depletion of Tudor-SN affects the association of Sm protein with snRNAs and, as a result, inhibits the assembly of uridine-rich small ribonucleoprotein mediated by the Sm core complex in vivo. Our results reveal the molecular basis for the involvement of Tudor-SN in regulating small nuclear ribonucleoprotein biogenesis, which provides novel insight related to the biological activity of Tudor-SN. PMID:22493508

  19. Staphylococcal cassette chromosome mec characterization of methicillin-resistant Staphylococcus aureus strains isolated at the military hospital of Constantine/Algeria.

    Science.gov (United States)

    Ouchenane, Z; Agabou, A; Smati, F; Rolain, J-M; Raoult, D

    2013-12-01

    Staphylococcal cassette chromosome mec is a genetic mobile element that carries the gene mecA mediating the methicillin resistance in staphylococci. The aim of this study is to type the Staphylococcal cassette chromosome mec (SCCmec) in 64 non-redundant methicillin-resistant Staphylococcus aureus (MRSA) strains recovered at the military hospital of Constantine (Algeria) between 2005 and 2007. Methicillin resistance was detected by oxacillin and cefoxitin discs and PBP2a test, and then confirmed by mecA PCR. The SCCmec complex types were determined by real time PCR. The analysis showed that 50 isolates were hospital acquired (HA-MRSA) and 14 were community-acquired (CA-MRSA). SCCmec type IV and V (traditionally attributed to CA-MRSA) were harbored by both HA-MRSA and CA-MRSA, while SCCmec type I, II and III were not recorded. These findings motivate more investigations to be carried on HA-MRSA in our hospital and other national health care centers.

  20. Nationwide German Multicenter Study on Prevalence of Antibiotic Resistance in Staphylococcal Bloodstream Isolates and Comparative In Vitro Activities of Quinupristin-Dalfopristin

    Science.gov (United States)

    von Eiff, Christof; Reinert, Ralf René; Kresken, Michael; Brauers, Johannes; Hafner, Dieter; Peters, Georg

    2000-01-01

    Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448 S. aureus isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the antistaphylococcal activities of quinupristin-dalfopristin were compared with those of eight other compounds by the broth microdilution method. The rates of oxacillin resistance in Staphylococcus aureus, S. epidermidis, S. haemolyticus, and other CoNS were 13.5, 69, 90, and 34%, respectively. In oxacillin-resistant strains high rates of resistance (up to 100%) to erythromycin, clindamycin, ciprofloxacin, and gentamicin were also observed. However, no strain appeared to be resistant to vancomycin or quinupristin-dalfopristin. The streptogramin combination exhibited excellent in vitro activity against all staphylococcal species tested, regardless of the patterns of resistance to other drug classes. In terms of MICs at which 90% of the isolates are inhibited, quinupristin-dalfopristin was 2 times more active against S. aureus isolates, 4 to 16 times more active against S. haemolyticus, and 8 to 32 times more active against S. epidermidis than vancomycin or teicoplanin. PMID:10921933

  1. Establishment of highly specific and quantitative immunoassay systems for staphylococcal enterotoxin A, B, and C using newly-developed monoclonal antibodies.

    Science.gov (United States)

    Sasaki, Takanori; Terano, Yoshitake; Shibata, Tadayoshi; Kawamoto, Hiroyoshi; Kuzuguchi, Tsuyoshi; Kohyama, Erina; Watanabe, Toshihiro; Ohyama, Tohru; Gemba, Munekazu

    2005-01-01

    Staphylococcal enterotoxin (SE) activities remain after boiling or treating with proteases. The main symptoms such as vomiting and diarrhea, are caused by the ingestion of SEs. Among SEs, SEA has been reported to be the major and most toxic protein. A highly specific and simple assay system is required to diagnose staphylococcal food poisoning. Therefore, the development of a suitable assay system is strongly anticipated. In this study, we have established a highly specific and sensitive avidin-biotin sandwich ELISA (ABS-ELISA) system for SEA, SEB, and SEC1 using newly-developed monoclonal antibodies. The linearity of these systems obtained was in the range of 0.78-25 ng/ml for each SE, and furthermore, the lower concentrations of SEs could also be detected. The recoveries of SEs from murine serum, skim milk solution, and raw milk were found to be over 90%, suggesting that our systems could detect SEs without any interventions, such as these from milk or serum proteins. We were also able to quantify SEs in 22 specimens of culture supernatants of S. aureus isolated in past occurrences. Our established system should be very useful not only in the clinical field but also in various fields of investigation because of its quantifi-cation and simplicity in detecting SEs.

  2. MerTK Does Not Mediate Phagocytosis of Staphylococcus aureus but Attenuates Inflammation Induced by Staphylococcal Lipoteichoic Acid Through Blocking NF-κB Activation.

    Science.gov (United States)

    Zhang, Bing; Wu, Huimei; Fang, Lei; Ding, Peishan; Xu, Ke; Yang, Qingbin; Liu, Rongyu

    2017-05-20

    Mer receptor tyrosine kinase (MerTK) expressed in macrophages is essential for phagocytosis of apoptotic cells. Here, we investigate whether MerTK is involved in the phagocytosis of Staphylococcus aureus (S. aureus) and regulation of staphylococcal lipoteichoic acid (LTA)-induced inflammatory response in macrophages. We found that stimulating RAW264.7 macrophages with S. aureus activated multiple signaling pathways including toll-like receptor 2 (TLR2), scavenger receptor A (SR-A), and MerTK. Meanwhile, S. aureus stimulation also induced activation of proteins focal adhesion kinase (FAK) and Rac1, which are related to phagocytosis. Pretreatment with a specific Mer-blocking antibody significantly inhibited S. aureus-induced phosphorylation of MerTK, while it had no effect on S. aureus-induced activation of FAK and Rac1. Moreover, by confocal laser microscope, we observed that the antibody blockade of MerTK had little impact on the phagocytosis of S. aureus by RAW264.7 macrophages. Additionally, pretreatment with this antibody further promoted LTA-induced phosphorylation of nuclear factor κB (NF-κB) p65 subunit and production of pro-inflammatory cytokines, such as TNF-α, IL-6, IL-1β, and macrophage inflammatory protein-2 (MIP-2). Collectively, these results suggest that MerTK does not play an essential role in the phagocytosis of S. aureus but attenuates inflammation induced by staphylococcal LTA through blocking NF-κB activation.

  3. The phage lytic proteins from the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 display multiple active catalytic domains and do not trigger staphylococcal resistance.

    Directory of Open Access Journals (Sweden)

    Lorena Rodríguez-Rubio

    Full Text Available The increase in antibiotic resistance world-wide revitalized the interest in the use of phage lysins to combat pathogenic bacteria. In this work, we analyzed the specific cleavage sites on the staphylococcal peptidoglycan produced by three phage lytic proteins. The investigated cell wall lytic enzymes were the endolysin LysH5 derived from the S. aureus bacteriophage vB_SauS-phi-IPLA88 (phi-IPLA88 and two fusion proteins between lysostaphin and the virion-associated peptidoglycan hydrolase HydH5 (HydH5SH3b and HydH5Lyso. We determined that all catalytic domains present in these proteins were active. Additionally, we tested for the emergence of resistant Staphylococcus aureus to any of the three phage lytic proteins constructs. Resistant S. aureus could not be identified after 10 cycles of bacterial exposure to phage lytic proteins either in liquid or plate cultures. However, a quick increase in lysostaphin resistance (up to 1000-fold in liquid culture was observed. The lack of resistant development supports the use of phage lytic proteins as future therapeutics to treat staphylococcal infections.

  4. Staphylococcal enterotoxins modulate interleukin 2 receptor expression and ligand-induced tyrosine phosphorylation of the Janus protein-tyrosine kinase 3 (Jak3) and signal transducers and activators of transcription (Stat proteins)

    DEFF Research Database (Denmark)

    Nielsen, M; Svejgaard, A; Röpke, C;

    1995-01-01

    Staphylococcal enterotoxins (SE) stimulate T cells expressing the appropriate variable region beta chain of (V beta) T-cell receptors and have been implicated in the pathogenesis of several autoimmune diseases. Depending on costimulatory signals, SE induce either proliferation or anergy in T cells...

  5. Research on the Effects and Mechanisms of Compound Radix Arnebiae Oil on Scald Wound%复方紫草油的烫伤治疗作用及其机制研究

    Institute of Scientific and Technical Information of China (English)

    张静; 冯琴喜; 夏清平; 李红; 袁晓环

    2012-01-01

    目的基金项目:黑龙江卫生厅科研课题(2007-0通过复方紫草油对烧烫伤大鼠治疗作用机制的研究,为临床针对性的治疗及护理提供基础资料.方法 将60只SD大鼠随机分为5组,各20只,正常对照组、单纯脱毛组、复方紫草油治疗组、京万红烫伤膏治疗组、自然恢复组.采用酶联免疫法测定血浆中脂质氧化、一氧化氮的含量变化.结果 复方紫草油治疗组、京万红烫伤膏治疗组、自然恢复组脂质氧化含量都明显升高(P<0.05);京万红烫伤膏治疗组、自然恢复组一氧化氮含量在烫伤后2、4 h逐渐增加,8 h达高峰,24 h后降低,差异有统计学意义(P<0.05);复方紫草油治疗组一氧化氮含量在烫伤前后波动较小;复方紫草油治疗组愈合时间少于京万红烫伤膏治疗组和自然恢复组,差异有统计学意义(P<0.05).结论 紫草油擦剂具有促进组织再生、抗菌消炎、止痛、抑制渗出等功效,能够参与组织的修复,加快组织的再生和愈合速度.%Objective To provide basic data for the clinical targeted therapy and nursing through study on the mechanisms of compound radix arnebiae oil to burn and scald of rats to. Methods 60 SD rats were randomly divided into 5 groups, 12 in each :control group, simple epilation group, compound radix arnebia oil group ,Jingwanhong ointment group, natural recovery group. Enzyme immunoassay method was used for the determination of plasma lipid oxidation and nitric oxide content. Results Lipid oxidation levels of compound radix arnebiae oil group, Jingwanhong ointment group, and natural recovery group were signiii- groupincreased gradually 2,4 h after burning,reaching the peak at 8 h,and declined after 24 h,the difference was statistically significant P <0. 05 ); nitric oxide content of the compound radix arnebiae oil group showed smaller fluctuation before and after binning; healing time of the group was shorter than than Jingwan-hong ointment

  6. Genes diferencialmente expressos em cana-de-açúcar inoculada com Xanthomonas albilineans, o agente causal da escaldadura da folha Diferential gene expression in sugar cane infected with Xanthomonas albilineans, causal agent of leaf scald

    Directory of Open Access Journals (Sweden)

    Karina Maia Dabbas

    2006-09-01

    Full Text Available A escaldadura da folha, causada pela bactéria Xanthomonas albilineans colonizadora do xilema, é uma das principais doenças da cana-de-açúcar. A sintomatologia na fase crônica é caracterizada principalmente pelo aparecimento de uma faixa branca paralela à nervura central da folha, que evolui até queimar totalmente, sendo também observado brotação de gemas laterais no colmo. Neste trabalho, a técnica de macroarranjos de cDNA foi empregada para o estudo da expressão de 3.575 ESTs (espressed sequence tags em folhas de cana-de-açúcar. Foram utilizadas duas variedades, uma resistente (SP82-1176 e outra suscetível (SP78-4467 a Xanthomonas albilineans as quais foram infectadas mecanicamente por ferimentos. As membranas dos macroarranjos foram confeccionadas a partir de ESTs de bibliotecas de folha e cartucho de cana-de-açúcar provenientes do projeto SUCEST e hibridizadas contra sondas de cDNA de plantas infectadas e controle marcadas com isótopos radioativos. Analisando os resultados dos macroarranjos foi possível verificar um comportamento diferenciado para cada variedade durante o ataque do patógeno. Após realizadas análises estatísticas identificamos na variedade resistente ESTs com expressão induzida relacionadas com biossíntese de isoprenoides, proteínas LRR transmembrânica, "ziper" de leucina, lignificação, tolerância ao frio, diferenciação de plastídeos, sistemas de defesa e de adaptação da planta ao meio ambiente. As ESTs reprimidas na variedade resistente foram àquelas relacionadas com genes responsáveis pela síntese de proteínas do controle da expansão da parede celular, detoxificação e transporte de auxina. Na variedade susceptível foram reprimidas ESTs relacionadas a genes de proteínas das respostas de defesa da planta, biossíntese de Etileno e regulação da transcrição.The leaf scald disease, caused by the xylem-invading pathogen Xanthomonas albilineans, is one of the most devastating

  7. Rapid detection of gene encoding staphylococcal enterotoxin B by PCR%葡萄球菌肠毒素B基因的PCR快速检测法

    Institute of Scientific and Technical Information of China (English)

    张勇; 石晓路; 梁伟; 李波; 陈润莉; 曾华书

    2013-01-01

    目的 建立快速检测葡萄球菌肠毒素B(staphylococcal enterotoxin B,SEB)基因的聚合酶链反应(PCR)方法.方法 ①根据SEB基因的序列,设计PCR引物特异性扩增靶基因片段.通过对1株产SEB金黄色葡萄球菌和9株对照菌株进行PCR检测,评价该方法的特异性;通过对产SEB金黄色葡萄球菌菌株做10倍系列稀释后进行PCR检测,评价该方法的敏感性;②分析30株金黄色葡萄球菌SEB基因的携带情况.结果 ①建立PCR方法快速检测SEB基因,扩增产物长度为494 bp.PCR反应体系中有26 CFU的产SEB金黄色葡萄球菌即可检出SEB基因;②30株分离的金黄色葡萄球菌中有2株携带有SEB基因.结论 PCR法可以快速、敏感地检测SEB基因,为金黄色葡萄球菌食物中毒诊断提供依据.%[Objective] To establish a PCR assay for the rapid and sensitive detection of Staphylococcal enterotoxin B (SEB) gene. [Methods]① According to the sequences of the SEB gene, primers were selected and were used to amplify SEB gene. To evaluate the specificity of the assay, 1 strain of SEB positive Staphylococcus aureus and 9 strains of non-Staphylococcus aureus were tested by PCR. To evaluate the sensitivity of the assay, the strain of SEB positive Staphylococcus aureus was 10-fold serially diluted and was amplified by PCR. ②The SEB gene in 30 strains of Staphylococcus aureus were detected by PCR. [Results] ①The PCR assay for the rapid and sensitive detection of SEB gene was well established. The length of amplification was 494 bp. The detection limit for SEB gene was 26 CFU;② Among 30 strains of Staphylococcus aureus, 2 strains were positive for SEB gene. [Conclusion]The PCR assay can rapidly and exactly detect the SEB gene, and can provide evidence for the rapid diagnosis of Staphylococcal food poisoning.

  8. 表皮剥脱毒素与桥粒芯糖蛋白在葡萄球菌性烫伤样皮肤综合征中的作用%The role of exfoliative toxin and desmogleins in staphylococcus aureus scalded skin syndrome

    Institute of Scientific and Technical Information of China (English)

    王珊珊; 路永红

    2014-01-01

    Exfoliative toxin (ET) produced by staphylococcus aureus coagulase positive phage group II ep-idermal type 71 acts on desmosomes and leads to the damage of junction of keratinocytes in staphylococcus aureus scalded skin syndrome (SSSS). The interaction between ET and desmoglein in SSSS is reviewed.%葡萄球菌性烫伤样皮肤综合征由金黄色葡萄球菌凝固酶阳性噬菌体 II 组71型产生的表皮剥脱毒素作用于桥粒引起角质细胞间桥连接受损所致。本文就葡萄球菌性烫伤样皮肤综合征中表皮剥脱毒素与桥粒芯糖蛋白之间的关系做一综述。

  9. 苯丙酸诺龙对烫伤模型大鼠雄激素受体介导靶基因转录调控的影响%Effect of nandrolone phenylpropionate on androgen receptor-mediated transcriptional regulation of target genes in rat scald models

    Institute of Scientific and Technical Information of China (English)

    李凯; 岑瑛

    2015-01-01

    BACKGROUND:Moderate to severe burn and trauma, treatment of which has been paid equal attention to wound surface, is always a difficulty of clinical systemic treatment and has a poor prognosis. Anabolic hormones have acquired secure and good results in the treatment of burns in both animals and clinical patients. Although use of anabolic hormones is restricted by the doping management, but its androgen receptor and nuclear receptor coregulators are the newly emerging areas of interests in the field of gene regulation mechanism in recent years. OBJECTIVE:To investigate the effect of nandrolone phenylpropionate on androgen receptor-mediated transcriptional regulation of target gene in rat scald models. METHODS: Thirty-six rats were randomly divided into nandrolone phenylpropionate, model and control groups. In the nandrolone phenylpropionate and model groups, rats were subjected to a 20% total body surface area second-degree scald injury by hot water. Two days after model preparation, rats in these two groups were intramuscularly injected with nandrolone phenylpropionate and saline, once every other day, for a total of 21 days. RESULTS AND CONCLUSION:The gene expression levels of steroid receptor coactivator-1 and insulin-like growth factor 1 in the rat livers and gonads (testes, ovaries) were significantly different between the nandrolone phenylpropionate and model groups (P 0.05)。说明在不同组织不同生理病理条件下苯丙酸诺龙对类固醇受体辅助活化因子1、c-myc、胰岛素样生长因子1基因表达的作用是不同的。

  10. STUDY OF ANTIMICROBIAL ACTIVITY OF NEW COMBINED OINTMENT ON THE MODEL OF PURULENT WOUND OF STAPHYLOCOCCAL ETIOLOGY

    Directory of Open Access Journals (Sweden)

    Tkachova O.V.

    2015-05-01

    Full Text Available Introduction. Prevalence of purulent pathologies and insufficient number of combined ointments with multi-effect on inflammation, infection and tissue damage are a prerequisite for scientists of NUPh to develop a new combined ointment composed of: antiseptics ethacridine lactate and thick chlorophyllipt extract, drug with reparative and anti-inflammatory activity – dexpanthenol, PEG-base of the IInd generation with the moderate dehydrating activity (180%. The range of pharmacological actions of Filetol ointment meets local therapy requirements in the IInd phase of the wound healing process. New ointment can also be applied in the Ist phase of wound healing process, if the wound contains a small amount of purulent exudate or does not contain it at all. Material & Methods. The purulent wound of staphylococcal etiology was reproduced in 24 white mature female mice weighing 14-17 g and subcutaneously injected with 0.2 ml of 10% solution of calcium chloride in the skin area, depilated beforehand, and after the development of necrosis on the 4th day injected with 0.2 ml of daily agar cultures S. aureus ATCC 25923 in the necrotizing area. In 48 hours of the infection, in the area of administration of staphylococcus cultures purulent necrotic wounds developed, which, depending on the size and intensity of the lesions, were subjected to reverse development in 9-15 days in a row. Animals were divided into three groups of 8 animals, including a positive control group, animals in which were not treated. In this study, the comparator was Levomecol ointment which is widely used in clinical practice for the local treatment of purulent wounds. Efficacy of treatment was evaluated on the basis of lesion value and intensity in study groups compared with the positive control, animal deaths in the case of generalization of the purulent process, quantitative dynamics of microorganisms in the wound and planimetric indicators (area of wounds and the percentage of

  11. Kinetics of staphylococcal opsonization, attachment, ingestion and killing by human polymorphonuclear leukocytes: a quantitative assay using [3H]thymidine labeled bacteria.

    Science.gov (United States)

    Verhoef, J; Peterson, P K; Quie, P G

    1977-01-01

    A method has been developed for studying quantitatively the separate processes of bacterial opsonization, phagocytosis, and killing by human polymorphonuclear leukocytes using [3H]thymidine labeled Staphylococcus aureus. Phagocytosis is determined by assaying for leukocytes-associated radioactivity after differential centrifugation and washing the leukocytes. Opsonization is studied by incubating bacteria with an opsonic source for varying durations and then adding leukocytes. By treatment of samples with the muralytic enzyme, lysostaphin, the attachment and ingestion phases of phagocytosis can be separated. Sampling for colony forming units after disruption of the leukocytes permits the measurement of bacterial killing. Using this method, differences in the kinetics of staphylococcal opsonization by normal and C2 deficient sera were defined, opsonic influences on the attachment and ingestion phases of pH agocytosis were delineated, and the influences of different opsonins and leukocyte populations on killing were determined.

  12. Construction of bifunctional molecules specific to antigen and antibody’s Fc-fragment by fusion of scFv-antibodies with staphylococcal protein A

    Directory of Open Access Journals (Sweden)

    Kolibo D. V.

    2009-06-01

    Full Text Available Aim. To develop approach for detection of scFv and their complexes with antigens. Methods. The fusion proteins, which include sequences of scFv and staphylococcal protein A, were constructed and the obtained bifunctional molecules were immunochemically analysed. Results. It was shown, that scFv fused with protein A and their complexes with antigens are effectively recognized by labelled immunoglobulins with unrestricted antigenic specificity. Conclusions. The fusion of scFv with protein A fragment is a perspective approach to increase the efficiency of application in ELISA. The obtained scFv, fused with protein A, could be used for development of test-systems for the detection of diphtheria toxin.

  13. Peanut allergy as a trigger for the deterioration of atopic dermatitis and precursor of staphylococcal and herpetic associated infections – case report

    Directory of Open Access Journals (Sweden)

    Dennis Ferreira

    2015-09-01

    Full Text Available Atopic dermatitis (AD is a multifactorial and chronic disease, with genetic, environmental, immunological and nutritional origins. AD may be aggravated by allergies associated with infections. This study aims to describe a paediatric case of AD in which the peanut allergy was the triggering factor to aggravate the disease, and was also the concomitant precursor of staphylococcal (methicillin-sensitive [i]Staphylococcus aureus[/i], carrier of the Panton-Valentine leukocidine (PVL genes and herpetic (Herpes Simplex – HSV infections. The clinical management approach and nursing strategies promoted a favourable evolution during the hospitalization period, besides the family approach, which was essential to control any flare-up of the disease. Adherence to a recommended diet and the use of strategies to prevent any recurrent infections were important to ensure the patient’s quality of life.

  14. Ultra-low power laser stimulation impairs the adhesion of Staphylococcus aureus to primary human cells, and interferes with the expression of staphylococcal pathogenic factors.

    Science.gov (United States)

    Petruzzelli, Sabina; Congiu, Antonio; Gallamini, Michele; Pompei, Raffaello

    2014-04-01

    Lasers are commonly used in several fields of medicine as a complementary therapy for internal medicine, surgery and also diagnostics. The efficacy of ultra-low level laser therapy (ULLLT) at power levels around 0.15 mW/cm(2) has been demonstrated both in in vitro experiments and in the clinical environment. This work used an ULLLT laser source to analyze its efficacy on Staphylococcus aureus adhesion to cells and on its ability to produce pathogenic factors. Laser stimulation succeeded in impairing the binding of S. aureus to primary human cells in culture and in inhibiting the expression of coagulase, one of the main staphylococcal pathogenic factors. The importance of the extracellular matrix (ECM) and the modification of the ECM redox potential in these activities were also evidenced.

  15. The Efficacy of the Quorum Sensing Inhibitor FS8 and Tigecycline in Preventing Prosthesis Biofilm in an Animal Model of Staphylococcal Infection

    Directory of Open Access Journals (Sweden)

    Andrea Giacometti

    2013-08-01

    Full Text Available We investigated the efficacy of tigecycline and FS8, alone or combined, in preventing prosthesis biofilm in a rat model of staphylococcal vascular graft infection. Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses followed by topical inoculation with 2 x 107 colony-forming units of Staphylococcus aureus, strain Smith diffuse. The study included a control group, a contaminated group that did not receive any antibiotic prophylaxis, and three contaminated groups that received: (i intraperitoneal tigecycline, (ii FS8-soaked graft, and (iii tigecycline plus FS8-soaked graft, respectively. Each group included 15 animals. The infection burden was evaluated by using sonication and quantitative agar culture. Moreover, an in vitro binding-study was performed to quantify the how much FS8 was coated to the surface of the prosthesis. Tigecycline, combined with FS8, against the adherent bacteria showed MICs (2.00 mg/L and MBCs (4.00 mg/L four-fold lower with respect to tigecycline alone in in vitro studies. The rat groups treated with tigecycline showed the lowest bacterial numbers (4.4 x 104 ± 1.2 x 104 CFU/mL. The FS8-treated group showed a good activity and significant differences compared to control group with bacterial numbers of 6.8 x 104 ± 2.0 x 104 CFU/mL. A stronger inhibition of bacterial growth was observed in rats treated with a combined FS8 and tigecycline therapy than in those that were singly treated with bacterial numbers of 101 CFU/mL graft. In conclusion, the ability to affect biofilm formation as well, its property to be an antibiotic enhancer suggests FS8 as alternative or additional agent to use in conjunction with conventional antimicrobial for prevention of staphylococcal biofilm related infection.

  16. Multiple-Strain Approach and Probabilistic Modeling of Consumer Habits in Quantitative Microbial Risk Assessment: A Quantitative Assessment of Exposure to Staphylococcal Enterotoxin A in Raw Milk.

    Science.gov (United States)

    Crotta, Matteo; Rizzi, Rita; Varisco, Giorgio; Daminelli, Paolo; Cunico, Elena Cosciani; Luini, Mario; Graber, Hans Ulrich; Paterlini, Franco; Guitian, Javier

    2016-03-01

    Quantitative microbial risk assessment (QMRA) models are extensively applied to inform management of a broad range of food safety risks. Inevitably, QMRA modeling involves an element of simplification of the biological process of interest. Two features that are frequently simplified or disregarded are the pathogenicity of multiple strains of a single pathogen and consumer behavior at the household level. In this study, we developed a QMRA model with a multiple-strain approach and a consumer phase module (CPM) based on uncertainty distributions fitted from field data. We modeled exposure to staphylococcal enterotoxin A in raw milk in Lombardy; a specific enterotoxin production module was thus included. The model is adaptable and could be used to assess the risk related to other pathogens in raw milk as well as other staphylococcal enterotoxins. The multiplestrain approach, implemented as a multinomial process, allowed the inclusion of variability and uncertainty with regard to pathogenicity at the bacterial level. Data from 301 questionnaires submitted to raw milk consumers were used to obtain uncertainty distributions for the CPM. The distributions were modeled to be easily updatable with further data or evidence. The sources of uncertainty due to the multiple-strain approach and the CPM were identified, and their impact on the output was assessed by comparing specific scenarios to the baseline. When the distributions reflecting the uncertainty in consumer behavior were fixed to the 95th percentile, the risk of exposure increased up to 160 times. This reflects the importance of taking into consideration the diversity of consumers' habits at the household level and the impact that the lack of knowledge about variables in the CPM can have on the final QMRA estimates. The multiple-strain approach lends itself to use in other food matrices besides raw milk and allows the model to better capture the complexity of the real world and to be capable of geographical

  17. Patient Characteristics, Management, and Predictors of Outcome from Severe Community-Onset Staphylococcal Sepsis in Northeast Thailand: A Prospective Multicenter Study.

    Science.gov (United States)

    West, T Eoin; Wikraiphat, Chanthiwa; Tandhavanant, Sarunporn; Ariyaprasert, Pitchayanant; Suntornsut, Pornpan; Okamoto, Shawna; Mahavanakul, Weera; Srisamang, Pramot; Phiphitaporn, Sunchai; Anukunananchai, Jirasak; Chetchotisakd, Ploenchan; Peacock, Sharon J; Chantratita, Narisara

    2017-02-06

    Staphylococcus aureus infection is a persistent threat in resource-restricted settings in southeast Asia but informative data about this disease remain limited. We analyzed characteristics, management, and predictors of outcome in severely septic patients with community-onset S. aureus infection in northeast Thailand. We performed a prospective, multicenter observational cohort study of community-onset S. aureus sepsis in four referral hospitals recruiting patients at least 14 years of age admitted between March 2010 and December 2013. One hundred and nineteen patients with severe staphylococcal sepsis were enrolled. Diabetes was the most common underlying condition. Methicillin-resistant infection was rare. Twenty-eight-day mortality was 20%. Ninety-two percent of patients received appropriate antibiotic therapy and 82% were administered intravenous fluids on the first hospital day, although only 14% were managed in an intensive care unit (ICU). On univariable analysis, clinical variables at enrollment significantly associated with death at 28 days were coagulopathy or respiratory failure. Plasma interleukin (IL) -8 concentration alone accurately predicted mortality (area under the receiver operating curve = 0.82, 95% confidence interval = 0.73-0.90). In multivariable analysis, addition of IL-8 concentration to a mortality prediction model containing clinical variables further improved the predictive ability of the model. We conclude that severe staphylococcal sepsis in northeast Thailand causes significant mortality. Diabetes is a common preexisting condition and most patients are managed outside the ICU even if they receive vasoactive/inotropic agents or mechanical ventilation. While clinical factors apparent on presentation including coagulopathy and respiratory failure predict death, plasma IL-8 improves this prediction.

  18. The optimal time for early excision in major burn injury.

    Science.gov (United States)

    Muangman, Pornprom; Sullivan, Stephen R; Honari, Shari; Engrav, Lorenz H; Heimbach, David M; Gibran, Nicole S

    2006-01-01

    Early excision and grafting (E&G) drastically changed burn care in America by reducing morbidity, mortality and hospital length of stay (LOS). The present study was intended to determine whether an optimal time window exists between resuscitation and wound sepsis for the first E&G in a patient with a large burn. The authors conducted a retrospective study of patients admitted between January 1994 and December 2000 with > or = 40% TBSA burns and at least 1 E&G procedure. Patients were grouped according to the day of their first operation. Patients allowed to heal indeterminate burns prior to excision and grafting of deep partial or full thickness burns were grouped as > or = d7 and were excluded from the present study. The authors correlated the time of first excision with infection, mortality and LOS. Seventy-five patients were identified and 12 patients allowed to heal indeterminate burn prior to excision and grafting of deep partial or full thickness burns were excluded. Sixty-three remaining patients included 51 males and 12 females. Mean burn size was 49% of total body surface area (TBSA) (44% deep partial or full thickness) and the mean age was 36 years. There were 61 flame (2 combined with electrical injuries), 1 scald and 1 chemical burn. Twelve died (19%) and 52 patients developed 121 infections. Whereas there was no statistical difference in mortality for patients operated on different days (p > 0.2), 60% of patients operated within the first 48 hours after injury died; this was not significant due to a small patient number The present data suggest that patients who undergo early excision and grafting within seven days following a major burn > or = 40% TBSA have equivalent infection or mortality rates regardless of when the first operation occurs between post burn day(PBD) 2 and PBD 7 (p > 0.2).

  19. An experimental research on anti-inflammatory effect of ecdysterone and bone mesenchymal stem cells at early period of burn wound%蜕皮甾酮和骨髓间充质干细胞对早期创面抗炎作用的初步研究

    Institute of Scientific and Technical Information of China (English)

    张子良; 吴旭; 张军花; 严智敏; 张元中; 冯长江; 付小兵

    2011-01-01

    Objective : To investigate the anti-inflammatory effect of ecdysterone (EDS) and bone mesenchymal stem cells (BMSC) at early period of burn wound in rabbits. Methods: Scald injury was reproduced in 18 rabbits by scald instrument, and 4 scald wounds of deep second degree were made on skin, and they were divided randomly to the groups of A, B, C and D. Wounds in group A was blank control group, group B was treated with EDS,group C with BMSC, and group D with BMSC+FDS. Six rabbits were sacrificed to obtain the histological specimens on 5, 10, 15 days (n= 6), respectively, after the treatment. The degree of infiltration of inflammatory cells, histopathological scores and wound healing rate were observed to assess the therapeutic effect. Results: Five days after scald, the degree of inflammatory cell infiltration of group B and D was lower than group A (P<0. 01) ,while no markedly changes were found in group C; the wound healing rate of group B was obviously higher than that of the group A (P<0. 05). On l0th day after scald. there were no significant differences among all groups in the degree of infiltration of inflammatory cells, wound healing rate and histopathological scores ( P> 0. 05). On 15th day after scald, there were no significant differences among all groups in the degree of infiltration of inflammatory cells (P>0. 05) , but the wound healing rates and histopathological scores of group B, C and D were obviously higher than that of group A (P<0. 05 or P<0. 01). Conclusion: FDS can enhance anti-inflammatory effect at an early period of scald wound in rabbits. It may be a mechanism of enhancing wound healing of FDS application. When EDS is combined with BMSC. the anti-inflammatory effect at early period of scald wound is enhanced.There is no antagonism between two agents, but the additive effects of them is not confirmed and warrants more research work.%目的:研究蜕皮甾酮(EDS)及骨髓间充质干细胞(BMSC)对烫伤创面的早期抗炎作

  20. Establishment of Indirect ELISA for Special Serum of Dairy-derived Staphylococcal enterotoxin A%乳源性金黄色葡萄球菌肠毒素A的间接ELISA检测方法建立

    Institute of Scientific and Technical Information of China (English)

    孙茂忠; 薛秀恒; 王菊花; 梅林; 吕高婧; 徐洪军

    2012-01-01

    Staphylococcal enterotoxin was collected and purified from Staphylococcus aureus,which was isolated from milk sample.In this way,a specific anti-serum of Staphylococcal enterotoxin A was prepared and the indirect ELISA method for detecting Staphylococcal enterotoxin was developed.The result showed that the optimal concentration of coating antigen is 10 μg/mL,the optimal dilution of anti-serum examined is 1︰2 000,ELISA titer is 1︰3 200.The foundation was provided for dairy-derived Staphylococcus aureus detection through this method.%从乳品中筛选出金黄色葡萄球菌,并对其所产的肠毒素进行分离纯化。制备出金黄色葡萄球菌肠毒素的特异性抗血清,建立金黄色葡萄球菌肠毒素A特异性血清间接ELISA方法。结果表明,最佳抗原包被浓度为10μg/mL,抗体最佳稀释倍数为1︰1 600,抗血清效价达到1︰3 200。通过建立此检测方法可以为乳源金黄色葡萄球菌肠毒素的检验提供依据。

  1. 金黄色葡萄球菌肠毒素B在变态反应性疾病发病机制中的作用%Effects of staphylococcal enterotoxin B in the pathogenesis of allergic disease

    Institute of Scientific and Technical Information of China (English)

    唐新业; 孙荣; 洪苏玲

    2011-01-01

    The prevalence of allergic disease increased over the last few decades. Type I hypersensitivity is the main pathogenesis of allergic disease. Staphylococcal enterotoxin B could act as allergen to induce allergic disease. In addition, staphylococcal enterotoxin B could act as superantigen to regulate the activity of immuno-modulatory and pro-inflammatory cell types, and play an important role on allergic disease. Here, we summarize the effects of staphylococcal enterotoxin B in the pathogenesis of allergic disease such as allergic rhinitis, asthma and atopic dermatitis.%变态反应性疾病的发病率逐年上升,Ⅰ型超敏反应是其主要的发病机制.近年来研究发现,金黄色葡萄球菌肠毒素B不但可以作为变应原诱导变态反应性疾病的发生,而且还可以作为超抗原影响免疫调节细胞和促炎细胞活性,在变态反应性疾病中发挥极其重要的作用.本文对金黄色葡萄球菌肠毒素B在变应性鼻炎、哮喘及特应性皮炎几种常见变态反应性疾病发生、发展中的作用机制作一综述.

  2. A highly sensitive fluorescence resonance energy transfer aptasensor for staphylococcal enterotoxin B detection based on exonuclease-catalyzed target recycling strategy

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Shijia; Duan, Nuo; Ma, Xiaoyuan; Xia, Yu; Wang, Hongxin; Wang, Zhouping, E-mail: wangzp@jiangnan.edu.cn

    2013-06-11

    Graphical abstract: -- Highlights: •An ultrasensitive FRET aptasensor was developed for staphylococcal enterotoxin B determination. •SEB was recognized by SEB aptamer with high affinity and specificity. •The Mn{sup 2+} doped NaYF{sub 4}:Yb/Er UCNPs used as donor to quencher dye (BHQ{sub 3}) in new FRET. •The fluorescence intensity was prominently amplified using an exonuclease-catalyzed target recycling strategy. -- Abstract: An ultrasensitive fluorescence resonance energy transfer (FRET) bioassay was developed to detect staphylococcal enterotoxin B (SEB), a low molecular exotoxin, using an aptamer-affinity method coupled with upconversion nanoparticles (UCNPs)-sensing, and the fluorescence intensity was prominently enhanced using an exonuclease-catalyzed target recycling strategy. To construct this aptasensor, both fluorescence donor probes (complementary DNA{sub 1}–UCNPs) and fluorescence quencher probes (complementary DNA{sub 2}–Black Hole Quencher{sub 3} (BHQ{sub 3})) were hybridized to an SEB aptamer, and double-strand oligonucleotides were fabricated, which quenched the fluorescence of the UCNPs via FRET. The formation of an aptamer–SEB complex in the presence of the SEB analyte resulted in not only the dissociation of aptamer from the double-strand DNA but also both the disruption of the FRET system and the restoration of the UCNPs fluorescence. In addition, the SEB was liberated from the aptamer–SEB complex using exonuclease I, an exonuclease specific to single-stranded DNA, for analyte recycling by selectively digesting a particular DNA (SEB aptamer). Based on this exonuclease-catalyzed target recycling strategy, an amplified fluorescence intensity could be produced using different SEB concentrations. Using optimized experimental conditions produced an ultrasensitive aptasensor for the detection of SEB, with a wide linear range of 0.001–1 ng mL{sup −1} and a lower detection limit (LOD) of 0.3 pg mL{sup −1} SEB (at 3σ). The fabricated

  3. Prevalence of staphylococcal enterotoxins, toxin genes and genetic-relatedness of foodborne Staphylococcus aureus strains isolated in the Marmara Region of Turkey.

    Science.gov (United States)

    Aydin, Ali; Sudagidan, Mert; Muratoglu, Karlo

    2011-08-02

    Staphylococcus aureus is a major foodborne pathogen and it has the ability to produce a number of extracellular toxins. We analyzed 1070 food samples obtained from retail markets and dairy farms in the Marmara Region of Turkey for the presence of S. aureus. Out of 147 isolates, 92 (62.6%) were enterotoxigenic. PCR was used to investigate the presence of staphylococcal enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu), exfoliative toxin genes (eta and etb) and the toxic-shock syndrome toxin gene (tst). The PCR results showed that 53.3% of the isolates contained staphylococcal enterotoxin-like (SEl) toxin genes (seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu) which were more frequent than classical enterotoxin genes (sea to see). Furthermore, seo, sei, sem, seg, seu and sec were found in 37.0, 32.7, 30.4, 29.3, 29.3 and 27.2% of the isolates, respectively. The tst gene was detected and confirmed by DNA sequencing in 9 isolates. The presence of eta and etb were not found in the isolates. Enterotoxigenic capabilities of isolates with SEA-SEE were investigated by ELISA. Enterotoxigenic S. aureus isolates produced one to three enterotoxins, with the most frequently produced types being enterotoxin A and C. There was a correlation of 72.1% between production of a specific toxin and the presence of the respective genes. PFGE analysis was used to identify genetic-relatedness of enterotoxigenic S. aureus isolates and the results revealed that 13 groups of isolates from different or the same origin that contained the same genes showed 100% homology with indistinguishable band patterns. The other enterotoxigenic isolates showed related band patterns with 72-86% homology in sea-, 61-90% homology in sec-, 80-96% homology in seh-, and 69-96% homology in sep-positive isolates. To our knowledge, this is the first study to examine enterotoxins and related gene contents of S. aureus food isolates in the Marmara

  4. Detection of gene encoding Staphylococcal enterotoxin A by PCR%PCR法检测葡萄球菌肠毒素A基因

    Institute of Scientific and Technical Information of China (English)

    张勇; 石晓璐; 周海涛; 李波; 陈润莉; 曾华书

    2012-01-01

    Objective To establish a polymerase chain reaction(PCR) assay for the rapid and sensitive detection of Staphylococcal enterotoxin A (SEA) gene. Methods According to the sequences of the SEA gene, primers were selected and were used to amplify SEA gene. The specificity of the assay was determined by detection of 1 SEA positive Staphylococcus aureus strain and 9 non-Staphylococcus aureus strains using PCR. The detection limit of the assay for SEA positive Staphylococcus aureus strain was 10-fold serially diluted and was amplified by PCR. The SEA gene in 30 Staphylococcus aureus strains were detected by PCR. Results The PCR assay for the rapid and sensitive detection of SEA gene was well established. The length of amplification was 439bp. The detection limit for SEA gene was 29 CFU. Among 30 Staphylococcus aureus strains 10 strains were positive for SEA gene. Conclusion The PCR assay can rapidly and exactly detect the SEA gene, and can provide evidence for the rapid diagnosis of food poisonings due to Staphylococcus aureus infection.%目的 建立PCR方法快速检测葡萄球菌肠毒素A(Staphylococcal enterotoxin A,SEA)基因.方法 根据SEA基因的序列,设计PCR引物特异性扩增靶基因片段,对1株产SEA金黄色葡萄球菌和9株对照菌株进行PCR检测,评价其特异性和敏感性.另检测30株金黄色葡萄球菌SEA基因的携带情况.结果 建立PCR方法快速检测SEA基因,扩增产物长度为439bp.PCR反应体系中有29cfu的产SEA金黄色葡萄球菌即可检出SEA基因,30株分离的金黄色葡萄球菌中有10株携带有SEA基因.结论 PCR法可以快速、敏感地检测SEA基因,为金黄色葡萄球菌食物中毒诊断提供依据.

  5. Reação de coaglutinação estafilocócica na identificação de micoplasmas Staphylococcal coagglutination for mycoplasma identification

    Directory of Open Access Journals (Sweden)

    Jorge Timenetsky

    1993-12-01

    Full Text Available A reação de coaglutinação estafilocócica foi utilizada como metodologia de identificação rápida de micoplasmas para ser aplicada em laboratórios não especializados. Amostras selvagens de micoplasmas isoladas de humanos, culturas celulares, ratos e camundongos foram identificados através da reação de coaglutinação estafilocócica utilizando-se do Staphylococcus aureus produtor de proteína A (amostra Cowan I sensibilizado com anticorpo de coelho contra amostra padrão micoplasma. Na identificação, os micoplasmas estavam em suspensão concentrada provenientes de 4,0 ml de cultivo. Quarenta e oito amostras de M.pulmonis, 6 de M. arthritidis, 8 de M.arginini, 3 de M.orale, 15 de A.laidlawii, 8 de M.hominis e 3 de M. pneumoniae foram identificadas pela coaglutinação estafilocócica e confirmadas pela inibição de crescimento. Parâmetros ótimos no preparo do conjugado e da reação de coaglutinação foram estabelecidos; o conjugado coaglutinante manteve-se estável por 90 dias quando adicionado com acetileisteína; a coaglutinação foi visualizada sem auxílio óptico. Os soros foram absorvidos com espécies padrões heterólogas e com o precipitado de caldo estéril.Staphylococcal Coagglutination was used as method for a rapid identification of mycoplasmas that could be performed by non specialized laboratories. Suspensions of Staphylococcus aureus (Cowan I sensitized with rabbit antibodies against NCTC mycoplasma strains have identified these microorganisms and the strains isolated from humans, cell cultures rats and mice in concentrated suspensions from cultures of 4.0 ml. Fourty eight strains of M.pulmonis, 6 of M. arthritidis, 8 of M.arginini, 3 of M.orale, 15 of A.laidlawii, 8 of M.hominis and 3 of M.pneumomae were identified by staphylococcal coagglutination and confirmed by Growth Inhibition Test. Optimal parameters of coagglutination were established and the stability of the conjugates were preserved for 90 days when

  6. 可调节封闭式负压引流技术联合局部氧疗对兔深Ⅱ度烫伤创面愈合的影响%Influence of Regulated Negative Pressure-Assisted Wound Therapy Combined with Topical Oxygen Therapy on the Healing of Deep Second Degree Scalded Rabbits

    Institute of Scientific and Technical Information of China (English)

    胡晓东; 李永忠; 李云飞; 陈莉; 蒋朔

    2015-01-01

    目的探讨可调节封闭式负压引流技术联合局部氧疗对兔深Ⅱ度烫伤创面愈合的影响,为临床应用提供实验依据。方法以40只新西兰大白兔为研究对象,建立兔深Ⅱ度烫伤模型,并将创伤兔随机分为A组(常规包扎治疗)、B组(局部氧疗)、C组(可调节封闭式负压引流治疗)、D组(可调节封闭式负压+局部氧疗),观察创面的大体变化及愈合情况,记录各组创面愈合率、愈合时间。结果 A组平均愈合天数是(25.2±2.1)d,B组平均愈合天数是(23.6±1.8)d,C组的平均愈合天数是(19.8±2.2)d,D组的平均愈合天数是(16.6±1.4)d,A组与其他三组在创面愈合时间差异上有统计学意义。治疗第12~18d,A组与其他三组在创面愈合率差异上有统计学意义。结论可调节封闭式负压引流技术与局部氧疗相结合能促进兔深Ⅱ度烫伤创面愈合,是一种新的治疗深Ⅱ度烫伤创面的方式。%Objective To probe into the influence of Negative Pressure-Assisted Wound Therapy (RNPT) combined with Topical Oxygen Therapy (TOT) on the healing of deep second degree scalded rabbits as to provide a experimental basis for clinical application. Methods Animal models of deep second degree scalded wound were established in 40 New Zealand rabbits, which were randomly divided into four groups:group A (routine enswathement), group B (TOT), group C (RNPT) and group D (RNPT +TOT).the changes in the wound were observed, Wound healing rate and time were respectively recorded. Results The wound healing time in group D was (16.6±1.4)d,which was significantly dif erent from those of the other groups, wound healing rate was also statistical y dif erent between group D and other groups from the 12th day to 18th day after treatment. Conclusion The new combined therapy of RNPT and TOT Can facilitate deep second degree scalded wound healing.

  7. Effects of rhGM-CSF on scalding wound healing and neovascularization in rats%重组人粒细胞巨噬细胞集落刺激因子对烫伤大鼠创面愈合及新生血管化的影响

    Institute of Scientific and Technical Information of China (English)

    刘继松; 方勇; 姚敏; 俞为荣; 杜娟

    2012-01-01

    Objective: To evaluate the effect of rhGM-CSF on scalding wound healing and neovascularization. Methods: Deep Ⅱ degree bum wound was made in72 SD rate on the back with diameter of3 cm. Rate were randomly divided into experinent group( n = 36, local wound was treated with rhGM-CSF gel) and control group(n =36, bcal wound were treated with gel matrix without rhGM-CSF). The process of wound healing was observed, and the percentages of wound closure were calculated on the day, the 1st,2 nd,3 rd, 7 th,10 th,14 th, and 21 st day after scald The expression of CD31, the surfacemaker of neovascularendothelial cells was detetcted with in the wound sites by immunoh istochamical staining, and the microvessel density was calculated. Results: The percentages of wound closure in experinent group were significantly higher than that in control group from the 7 th day to the21 st day. Immunohistochanical detection revealed that the expression of CD31 and the microvessel densily in experinent group were significantly higher than those in control group from7 th day to21 stday(P <0.01). Conclusions: The treatment of rhGM-CSF may promote scalding wound healing in rate The mechanisn may be associated with upregulation of CD31 expression and acceleration of neovascularization.%目的:研究重组人粒细胞巨噬细胞集落刺激因子(recombinant human granulocyte-macrophage colony-stimulating factor,rhGM-CSF)对烫伤创面组织愈合及新生血管化的影响.方法:72只SD大鼠于背部制备直径3 cm的深Ⅱ度烫伤创面.根据创面不同处理方式将大鼠随机分为实验组(创面局部应用rhGM-CSF凝胶剂)和对照组(创面局部应用不含rhGM-CSF的凝胶基质),每组36只.分别于创面形成后当日和第1、3、7、10、14、21天观察创面并计算创面愈合率;免疫组织化学染色观察创面组织中新生血管内皮细胞表面标志物CD31表达,计算微血管密度.结果:自创面形成后第7天起,实验组创面愈

  8. Effect of Boschniakia rossica on expression of GST-P, p53 and p21(ras)proteins in early stage of chemical hepatocarcinogenesis and its anti-inflammatory activities in rats.

    Science.gov (United States)

    Yin, Zong-Zhu; Jin, Hai-Ling; Yin, Xue-Zhe; Li, Tian-Zhu; Quan, Ji-Shu; Jin, Zeng-Nan

    2000-12-01

    AIM:To investigate the effect of Boschniakia rossica (BR) extract on expression of GST-P, p53 and p21(ras) proteins in early stage of chemical hepatocarcinogenesis in rats and its anti-inflammatory activities.METHODS:The expression of tumor marker-placental form glutathione S-transferase (GST-P), p53 and p21(ras) proteins were investigated by immunohisto-chemical techniques and ABC method. Anti-inflammatory activities of BR were studied by xylene and croton oil-induced mouse ear edema, carrageenin, histamine and hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet induced mouse granuloma formation methods.RESULTS:The 500mg/kg of BR-H2O extract frac-tionated from BR-Methanol extract had inhibitory effect on the formation of DEN-induced GST-P-positive foci in rat liver (GST-P staining was 78% positive in DEN+AAF group vs 20% positive in DEN+AAF+BR group, Panti-inflamatory effect in xylene and crotonoil induced mouse ear edema (inhibitory rates were 26%-29% and 35%-59%, respectively). BR H(2)O extract exhibited inhibitory effect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in rats and cotton pellet-induced granuloma formation in mice.CONCLUSION:BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in early stage of rat chemical hepato-carcinogenesis.Both CH(2)Cl(2) and H(2)O extracts from BR exerted anti-inflammatory effect in rats and mice.

  9. Early clerkships

    NARCIS (Netherlands)

    Kamalski, Digna M. A.; Ter Braak, Edith W. M. T.; Ten Cate, Olle Th. J.; Borleffs, Jan C. C.

    2007-01-01

    Background: Early clinical experience is being introduced in innovative, vertically integrated undergraduate medical curricula. While in many cases, this early clinical experience is limited to the presence of patients during lectures, in Utrecht students gain 'hands on' experience of daily clinical

  10. Effects of dragon's blood on the expression of substance P and Bcl-2 in wound tissue of diabetic scalded rats%糖尿病模型大鼠烫伤创面皮肤组织P物质和Bcl-2表达与龙血竭的干预

    Institute of Scientific and Technical Information of China (English)

    张宪发; 吴正球; 梁自乾; 赵学凯; 丁华荣

    2012-01-01

    BACKGROUND: Previous studies have demonstrated that dragon's blood can promote the wound healing as well as have characteristics of anti-platelet aggregation, anti-inflammatory analgesic, anti-bacterial and antioxidant and other biological activity. OBJECTIVE: To identify the effects of dragon's blood on the expression of substance P and Bcl-2 in wound tissue of diabetic scalded rats.METHODS: A total of 112 Wistar rats were randomly divided into four groups: dragon's blood group, silver sulfadiazine (SD-Ag) group, diabetic control group and normal control group. Diabetic deep II degree burn models were established in dragon's blood group, SD-Ag group and diabetic control group, and the normal deep II degree burn models were established in normal control group.RESULTS AND CONCLUSION: On the 7th day post scald, the wound healing rates of dragon's blood group and normal control group were higher than those of SD-Ag group and diabetic control group (P 0.05). The results demonstrate that dragon's blood can promote the healing of diabetic burn wounds significantly by regulating the expression of substance P and Bcl-2.%背景:研究证明龙血竭有促进创伤愈合、抗血小板聚集、抗炎镇痛、抗菌抗氧化等生物活性.目的:验证龙血竭在糖尿病大鼠烫伤皮肤组织愈合过程中对P物质和Bcl-2表达的影响.方法:Wistar大鼠112只,随机分为4组.龙血竭组、磺胺嘧啶银(SD-Ag)组、糖尿病对照组和正常对照组.其中前3组为糖尿病深Ⅱ度烫伤模型,正常对照组为正常深Ⅱ度烫伤模型.结果与结论:烫伤后第7天开始创面愈合率龙血竭组、正常对照组高于SD-Ag组和糖尿病对照组(P 0.05).说明龙血竭能通过参与调控P物质和Bcl-2的表达,有效地促进糖尿病烫伤创面的愈合.

  11. Photodynamic antibacterial enhanced effect of methylene blue-gold nanoparticles conjugate on Staphylococcal aureus isolated from impetigo lesions in vitro study.

    Science.gov (United States)

    Tawfik, Abeer Attia; Alsharnoubi, Jehan; Morsy, Mona

    2015-06-01

    Staphylococcal aureus is the most common organism which has been encountered in impetigo infection. Gold nanoparticles can be used as a tool to deliver antimicrobials or to enhance photodynamic destruction of bacteria. To evaluate the photodynamic effect of methylene blue gold nanoparticles (MB-gold nanoparticles conjugate) on S. aureus which were isolated from impetigo lesions. Twenty children were diagnosed clinically as impetigo, and aged from 3 to 5 years of both sexes were recruited in the study. Two bacteriological samples were collected from each patient, identified and cultured. Samples of S. aureus of a concentration of 10(-1)ml were divided into four groups. S. aureus was treated by MB-gold nanoparticles conjugate, gold nanoparticles, MB, and the fourth group served as a control group. Diode laser (660 nm) was used for photoactivation. The bacterial growth inhibition was determined by two methods: the percentage of reduction of viable bacteria count and the optical density (O.D) of bacterial growth. The highest significant inhibitory effect on S. aureus was obtained with MB-gold nanoparticles conjugate when irradiated by diode laser 660 nm (P methylene blue. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Systematic survey on the prevalence of genes coding for staphylococcal enterotoxins SElM, SElO, and SElN.

    Science.gov (United States)

    Becker, Karsten; Friedrich, Alexander W; Peters, Georg; von Eiff, Christof

    2004-12-01

    Staphylococcus aureus remains a leading cause of food-poisoning with substantial impact on public health. Using a multiplex polymerase chain reaction-DNA enzyme immunoassay (PCR-DEIA), we studied the presence of genes encoding staphylococcal enterotoxin-like (SEl) superantigens sem, sen, and seo, associated with the enterotoxin gene cluster (egc), in 429 clinical Staphylococcus aureus isolates. 294 (68.5%) isolates tested positive for at least one of the three SEl genes. In contrast to the fixed gene combination seg/sei also located on egc, a substantial number of isolates (n = 108) were found to bear only one or two of the genes encoding SElM, SElN, and SElO. Regarding the origin of the S. aureus isolates, a significant difference (P = 0.022) was found for the possession of seo (61.2% of blood isolates versus 42.9% of nasal strains). Also sem (not significantly) was found more common in blood isolates (52.1% versus 40.5%). The survey of the newly described SEl genes sem-seo supports the concept that most clinical S. aureus isolates harbor subsets of pyrogenic toxin superantigens. The potential contribution of seo and sem to the pathogenic potential of S. aureus has to be further evaluated.

  13. Study of pK values and effective dielectric constants of ionizable residues in pentapeptides and in staphylococcal nuclease (SNase) using a mean-field approach.

    Science.gov (United States)

    Bossa, Guilherme Volpe; Fahr, Alfred; Pereira de Souza, Tereza

    2014-04-17

    The determination of pK values of amino acid residues as a function of temperature and ionic concentration is crucial to understanding the dynamics of various biological processes such as adsorption of peptides and their interactions with active sites of enzymes. In this study we developed a mean-field model to calculate the position-dependent dielectric constants of ionizable groups and the mean electrostatic potential on the surface. Such potential, which takes into account the contributions exerted by neighboring groups and ions in solution, is responsible for the fine-tuning of the pK value of each residue. The proposed model was applied to the amino acids Asp, Glu, Lys, His, Tyr, and Cys, and since the results were consistent with experimentally obtained values, the model was extended and applied to computation of pK values of Gly and Ala pentapeptides and of ionizable residues of the enzyme staphylococcal nuclease (SNase). In this latter case, we used an approach similar to a first-neighbors approximation, and the results turned out to be in good agreement with previously reported data when considering only the interactions of charged groups located at distances of maximally 20 Å. These considerations and the little computational cost involved turn the suggested approach into a promising tool for the modeling of force fields in computational simulations.

  14. Chimeric anti-staphylococcal enterotoxin B antibodies and lovastatin act synergistically to provide in vivo protection against lethal doses of SEB.

    Directory of Open Access Journals (Sweden)

    Mulualem E Tilahun

    Full Text Available Staphylococcal enterotoxin B (SEB is one of a family of toxins secreted by Staphylococcus aureus that act as superantigens, activating a large fraction of the T-cell population and inducing production of high levels of inflammatory cytokines that can cause toxic shock syndrome (TSS and death. Extracellular engagement of the TCR of T-cells and class II MHC of antigen presenting cells by SEB triggers the activation of many intracellular signaling processes. We engineered chimeric antibodies to block the extracellular engagement of cellular receptors by SEB and used a statin to inhibit intracellular signaling. Chimeric human-mouse antibodies directed against different neutralizing epitopes of SEB synergistically inhibited its activation of human T-cells in vitro. In the in vivo model of lethal toxic shock syndrome (TSS in HLA-DR3 transgenic mice, two of these antibodies conferred significant partial protection when administered individually, but offered complete protection in a synergistic manner when given together. Similarly, in vivo, lovastatin alone conferred only partial protection from TSS similar to single anti-SEB antibodies. However, used in combination with one chimeric neutralizing anti-SEB antibody, lovastatin provided complete protection against lethal TSS in HLA-DR3 transgenic mice. These experiments demonstrate that in vivo protection against lethal doses of SEB can be achieved by a statin of proven clinical safety and chimeric human-mouse antibodies, agents now widely used and known to be of low immunogenicity in human hosts.

  15. Evaluation of the VIDAS staph enterotoxin II (SET 2) immunoassay method for the detection of staphylococcal enterotoxins in selected foods: collaborative study.

    Science.gov (United States)

    Jechorek, Robert P; Johnson, Ronald L

    2008-01-01

    A multilaboratory study was conducted to determine the limit of detection (LOD) of Staphylococcal enterotoxins (SET) in 5 foods. Cooked chicken, ham, potato salad, pasteurized liquid whole milk, and canned mushrooms were each spiked with a different enterotoxin (A, B, C1, D, or E), and tested at 0.25 and 0.5 ng/g SET levels to determine the LOD of the assay for those foods in a collaborative study. Unspiked controls were also included. A total of 19 laboratories representing government and industry participated. In this study, 1674 test portions were analyzed, of which 1638 were used in the statistical analysis. Of the 1638 test portions used in the statistical analysis, 1104 were spiked test portions, of which 1073 were positive by the VIDAS Staph enterotoxin II (SET 2) method. The detection rates at the 0.25 ng/mL level were cooked chicken, 98.2%; ham, 99.0%; potato salad, 99.1%; liquid whole milk, 85.2%; and canned mushrooms, 100%. The detection rates at the 0.5 ng/mL level were cooked chicken, 97.4%; ham, 98.1%; potato salad, 100%; liquid whole milk, 99.0%; and canned mushrooms, 100%. The data indicate that the SET 2 method is capable of detecting SET at 0.25 ng/g in cooked chicken, ham, potato salad, and canned mushrooms and at 0.5 ng/g in pasteurized liquid whole milk.

  16. Highly efficient production of the staphylococcal nuclease reporter in Lactobacillus bulgaricus governed by the promoter of the hlbA gene.

    Science.gov (United States)

    Chouayekh, Hichem; Serror, Pascale; Boudebbouze, Samira; Maguin, Emmanuelle

    2009-04-01

    Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) genome sequence analysis revealed the presence of two genes that encode histone-like HU proteins (hlbA and hlbB) showing extensive similarity to other bacterial homologues. These genes were found to be extremely conserved among several L. bulgaricus strains. The hlbA gene was shown to be constitutively transcribed from a unique promoter (phlbA) during normal growth, whereas hlbB did not seem to be expressed under usual laboratory conditions. Using a reporter cassette in which the staphylococcal nuclease was fused at its N-terminus to the lactococcal signal peptide Usp45 (SP Usp45), we have demonstrated that phlbA promotes high expression of the reporter in L. bulgaricus, which correlated with an abundant secretion of the mature nuclease in the supernatant fraction. Quantification of the exported enzyme reveals a secretion level approximately threefold higher when the expression of the reporter was under the control of phlbA compared with the lactococcal usp45 promoter. Together, these results indicate that phlbA is suitable for gene expression in L. bulgaricus, that SP Usp45 is functionally recognized and processed by the L. bulgaricus secretion machinery and that the nuclease reporter gene can be used for the identification of exported products in this bacterium.

  17. Study on interactions of human IgG with immobilized anti-IgG or recombinant Staphylococcal protein A using surface plasmon resonance spectrometry

    Directory of Open Access Journals (Sweden)

    Bakhmachuk A. O.

    2016-02-01

    Full Text Available Aim. Comparison of the IgG-binding activity of recombinant Staphylococcal protein A with introduced C-terminal cysteine residue (SPA-Cys or goat anti-human IgG antibodies (anti-IgG after their immobilization on a gold sensor surface of surface plasmon resonance (SPR spectrometer. Methods. SPA-Cys or anti-IgG were immobilized on a gold sensor surface to form two variants of a bioselective element of the immunosensor. SPR spectrometry was used for the detection of IgG-binding activity of the immobilized proteins. Results.The SPR sensor response to the immobilization of anti-IgG was more than two times higher than that at the immobilization of SPA-Cys. However, there is almost the double advantage for SPA-Cys in the number of immobilized molecules. Moreover, the bioselective element of the immunosensor based on SPA-Cys showed a much better capability of binding Ig than bioselective element based on anti-IgG. Conclusions.The study on the immobilization of SPA-Cys or anti-IgG on the sensor surface of SPR spectrometer, and the interactions of immobilized proteins with human IgG demonstrated obvious advantages of SPA-Cys.

  18. Investigation of a staphylococcal food poisoning outbreak combining case-control, traditional typing and whole genome sequencing methods, Luxembourg, June 2014.

    Science.gov (United States)

    Mossong, Joël; Decruyenaere, Frédéric; Moris, Gilbert; Ragimbeau, Catherine; Olinger, Christophe M; Johler, Sophia; Perrin, Monique; Hau, Patrick; Weicherding, Pierre

    2015-01-01

    In June 2014, a staphylococcal food poisoning outbreak occurred at an international equine sports event in Luxembourg requiring the hospitalisation of 31 persons. We conducted a microbiological investigation of patients and buffet items, a case-control study and a carriage study of catering staff. Isolates of Staphylococcus aureus from patients, food and catering staff were characterised and compared using traditional typing methods and whole genome sequencing. Genotypically identical strains (sequence type ST8, spa-type t024, MLVA-type 4698, enterotoxin A FRI100) were isolated in 10 patients, shiitake mushrooms, cured ham, and in three members of staff. The case-control study strongly suggested pasta salad with pesto as the vehicle of infection (pfood item could not be tested, because there were no leftovers. Additional enterotoxigenic strains genetically unrelated to the outbreak strain were found in four members of staff. Non-enterotoxigenic strains with livestock-associated sequence type ST398 were isolated from three food items and two members of staff. The main cause of the outbreak is likely to have been not maintaining the cold chain after food preparation. Whole genome sequencing resulted in phylogenetic clustering which concurred with traditional typing while simultaneously characterising virulence and resistance traits.

  19. Staphylococcal superantigen-like genes, ssl5 and ssl8, are positively regulated by Sae and negatively by Agr in the Newman strain.

    Science.gov (United States)

    Pantrangi, Madhulatha; Singh, Vineet K; Wolz, Christiane; Shukla, Sanjay K

    2010-07-01

    Some of the staphylococcal superantigen-like (SSL) proteins SSL5, SSL7, SSL9, and SSL11 act as immunomodulatory proteins in Staphylococcus aureus. However, little is known about their regulatory mechanisms. We determined the expression levels of ssl5 and ssl8 in seven clinically important S. aureus strains and their regulatory mechanisms in the Newman strain, which had the highest ssl5 and ssl8 expression. Independent comparisons of ssl5 or ssl8 coding and upstream sequences in these strains identified multiple haplotypes that did not correlate with the differential expression of ssl5 and ssl8, suggesting the role of additional regulatory elements. Using knockout mutant strains of known S. aureus global regulators such as Agr, Sae, and SigB in the Newman strain, we showed that both ssl5 and ssl8 were induced by Sae and repressed by Agr, suggesting that Sae and Agr are the positive and the negative regulators, respectively, of these two ssl genes. Moreover, we observed upregulation of sae in the agr mutant and upregulation of agr in the sae mutant compared with the isogenic Newman strain, suggesting that the Agr and Sae may be inhibiting each other. The SigB mutation did not affect ssl5 and ssl8 expression, but they were downregulated in the agr/sigB double mutant, indicating that SigB probably acts synergistically with Agr in their upregulation.

  20. Transcription of the toxin genes present within the Staphylococcal phage phiSa3ms is intimately linked with the phage's life cycle.

    Science.gov (United States)

    Sumby, Paul; Waldor, Matthew K

    2003-12-01

    phiSa3ms, a lysogenic bacteriophage encoding the staphylococcal enterotoxins SEA, SEG, and SEK and the fibrinolytic enzyme staphylokinase (Sak), was identified in the unannotated genome sequence of the hypervirulent community-acquired Staphylococcus aureus strain 476. We found that mitomycin C induction of phiSa3ms led to increased transcription of all four virulence factors. The increase in sea and sak transcription was a result of read-through transcription from upstream latent phage promoters and an increase in phage copy number. The majority of the seg2 and sek2 transcripts were shown to initiate from the upstream phage cI promoter and hence were regulated by factors influencing cI transcription. The lysogeny module of phiSa3ms was shown to have some lambda-like features with divergent cI and cro genes. Band shift assays were used to identify binding sites for both CI and Cro within the region between these genes, suggesting a mechanism of control for the phiSa3ms lytic-lysogenic switch. Our findings suggest that the production of phage-encoded virulence factors in S. aureus may be regulated by processes that govern lysogeny.

  1. Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA.

    Science.gov (United States)

    Djoudi, Ferhat; Bonura, Celestino; Touati, Abdelaziz; Aléo, Aurora; Benallaoua, Said; Mammina, Caterina

    2016-11-01

    Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcushaemolyticus, 7 Staphylococcusaureus, 4 Staphylococcussciuri, 2 Staphylococcussaprophyticus and 1 isolate each of Staphylococcusxylosus and Staphylococcuslentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new β-lactam resistance.

  2. Antibacterial activity and therapeutic efficacy of Fl-PRPRPL-5, a cationic amphiphilic polyproline helix, in a mouse model of staphylococcal skin infection

    Directory of Open Access Journals (Sweden)

    Thangamani S

    2015-10-01

    Full Text Available Shankar Thangamani,1 Manish Nepal,2 Jean Chmielewski,2 Mohamed N Seleem11Department of Comparative Pathobiology, Purdue University College of Veterinary Medicine, West Lafayette, IN, USA; 2Department of Chemistry, Purdue University, West Lafayette, IN, USAAbstract: The antibacterial activities and therapeutic efficacy of the cationic, unnatural proline-rich peptide Fl-PRPRPL-5 were evaluated against multidrug-resistant Staphylococcus aureus in a mouse model of skin infection. Fl-PRPRPL-5 showed potent activity against all clinical isolates of S. aureus tested, including methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA, respectively. Fl-PRPRPL-5 was also superior in clearing established in vitro biofilms of S. aureus and Staphylococcus epidermidis, compared with the established antimicrobials mupirocin and vancomycin. Additionally, topical treatment of an MRSA-infected wound with Fl-PRPRPL-5 enhanced wound closure and significantly reduced bacterial load. Finally, 0.5% Fl-PRPRPL-5 significantly reduced the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α, interleukin-6 (IL-6, and interleukin-1 beta (IL-1β in wounds induced by MRSA skin infection. In conclusion, the results of this study suggest the potential application of Fl-PRPRPL-5 in the treatment of staphylococcal skin infections.Keywords: antimicrobial peptides, Staphylococcus aureus, biofilms, anti-inflammatory, skin infection

  3. Antibacterial activity and therapeutic efficacy of Fl-P(R)P(R)P(L)-5, a cationic amphiphilic polyproline helix, in a mouse model of staphylococcal skin infection.

    Science.gov (United States)

    Thangamani, Shankar; Nepal, Manish; Chmielewski, Jean; Seleem, Mohamed N

    2015-01-01

    The antibacterial activities and therapeutic efficacy of the cationic, unnatural proline-rich peptide Fl-P(R)P(R)P(L)-5 were evaluated against multidrug-resistant Staphylococcus aureus in a mouse model of skin infection. Fl-P(R)P(R)P(L)-5 showed potent activity against all clinical isolates of S. aureus tested, including methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA, respectively). Fl-P(R)P(R)P(L)-5 was also superior in clearing established in vitro biofilms of S. aureus and Staphylococcus epidermidis, compared with the established antimicrobials mupirocin and vancomycin. Additionally, topical treatment of an MRSA-infected wound with Fl-P(R)P(R)P(L)-5 enhanced wound closure and significantly reduced bacterial load. Finally, 0.5% Fl-P(R)P(R)P(L)-5 significantly reduced the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) in wounds induced by MRSA skin infection. In conclusion, the results of this study suggest the potential application of Fl-P(R)P(R)P(L)-5 in the treatment of staphylococcal skin infections.

  4. Antibacterial activity and therapeutic efficacy of Fl-PRPRPL-5, a cationic amphiphilic polyproline helix, in a mouse model of staphylococcal skin infection

    Science.gov (United States)

    Thangamani, Shankar; Nepal, Manish; Chmielewski, Jean; Seleem, Mohamed N

    2015-01-01

    The antibacterial activities and therapeutic efficacy of the cationic, unnatural proline-rich peptide Fl-PRPRPL-5 were evaluated against multidrug-resistant Staphylococcus aureus in a mouse model of skin infection. Fl-PRPRPL-5 showed potent activity against all clinical isolates of S. aureus tested, including methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA, respectively). Fl-PRPRPL-5 was also superior in clearing established in vitro biofilms of S. aureus and Staphylococcus epidermidis, compared with the established antimicrobials mupirocin and vancomycin. Additionally, topical treatment of an MRSA-infected wound with Fl-PRPRPL-5 enhanced wound closure and significantly reduced bacterial load. Finally, 0.5% Fl-PRPRPL-5 significantly reduced the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) in wounds induced by MRSA skin infection. In conclusion, the results of this study suggest the potential application of Fl-PRPRPL-5 in the treatment of staphylococcal skin infections. PMID:26543355

  5. Reduction of Non-Specific Protein Adsorption Using Poly(ethylene Glycol (PEG Modified Polyacrylate Hydrogels In Immunoassays for Staphylococcal Enterotoxin B Detection

    Directory of Open Access Journals (Sweden)

    Paul T. Charles

    2009-01-01

    Full Text Available Three PEG molecules (PEG-methacrylate, -diacrylate and -dimethacrylate were incorporated into galactose-based polyacrylate hydrogels and their relative abilities to reduce non-specific protein adsorption in immunoassays were determined. Highly crosslinked hydrogels containing amine-terminated functionalities were formed and used to covalently attach antibodies specific for staphylococcal enterotoxin B (SEB. Patterned arrays of immobilized antibodies in the PEG-modified hydrogels were created with a PDMS template containing micro-channels for use in sandwich immunoassays to detect SEB. Different concentrations of the toxin were applied to the hydrogel arrays, followed with a Cy3-labeled tracer antibody specific for the two toxins. Fluorescence laser scanning confocal microscopy of the tracer molecules provided both qualitative and quantitative measurements on the detection sensitivity and the reduction in non-specific binding as a result of PEG incorporation. Results showed the PEG-modified hydrogel significantly reduced non-specific protein binding with a detection limit for SEB of 1 ng/mL. Fluorescence signals showed a 10-fold decrease in the non-specific binding and a 6-fold increase in specific binding of SEB.

  6. GPVI and GPIbα mediate staphylococcal superantigen-like protein 5 (SSL5 induced platelet activation and direct toward glycans as potential inhibitors.

    Directory of Open Access Journals (Sweden)

    Houyuan Hu

    Full Text Available BACKGROUND: Staphylococcus aureus (S. aureus is a common pathogen capable of causing life-threatening infections. Staphylococcal superantigen-like protein 5 (SSL5 has recently been shown to bind to platelet glycoproteins and induce platelet activation. This study investigates further the interaction between SSL5 and platelet glycoproteins. Moreover, using a glycan discovery approach, we aim to identify potential glycans to therapeutically target this interaction and prevent SSL5-induced effects. METHODOLOGY/PRINCIPAL FINDINGS: In addition to platelet activation experiments, flow cytometry, immunoprecipitation, surface plasmon resonance and a glycan binding array, were used to identify specific SSL5 binding regions and mediators. We independently confirm SSL5 to interact with platelets via GPIbα and identify the sulphated-tyrosine residues as an important region for SSL5 binding. We also identify the novel direct interaction between SSL5 and the platelet collagen receptor GPVI. Together, these receptors offer one mechanistic explanation for the unique functional influences SSL5 exerts on platelets. A role for specific families of platelet glycans in mediating SSL5-platelet interactions was also discovered and used to identify and demonstrate effectiveness of potential glycan based inhibitors in vitro. CONCLUSIONS/SIGNIFICANCE: These findings further elucidate the functional interactions between SSL5 and platelets, including the novel finding of a role for the GPVI receptor. We demonstrate efficacy of possible glycan-based approaches to inhibit the SSL5-induced platelet activation. Our data warrant further work to prove SSL5-platelet effects in vivo.

  7. Early literacy

    DEFF Research Database (Denmark)

    Jensen, Anders Skriver

    2012-01-01

    This paper discusses findings from the Danish contribution to the EASE project, a European research project running from 2008 to 2010 on early literacy in relation to the transition from childcare to school. It explores a holistic, inclusive approach to early literacy that resists a narrow...... and schools. The paper also draws on Gee’s (2001, 2003, 2004, 2008) sociocultural approach to literacy, and Honneth’s (2003, 2006) concept of recognition. Emphasizing participation and recognition as key elements, it claims that stakeholders in early liter- acy must pay attention to how diverse early literacy...... opportunities empower children, especially when these opportunities are employed in a project-based learning environ- ment in which each child is able to contribute to the shared literacy events....

  8. Effect of recombinant human granulocyte-macrophage colony stimulating factor gel on melting crust in deep partial thickness scalding rats%GM-CSF 凝胶对 SD 大鼠深 II度烧伤创面溶痂的影响

    Institute of Scientific and Technical Information of China (English)

    杜娟; 刘继松; 章祥洲; 赵经伟; 方勇; 姚敏; 俞为荣

    2014-01-01

    目的:观察重组人粒细胞巨噬细胞集落刺激因子( rhGM-CSF)对深II度烧伤创面坏死组织溶痂和促进创面愈合的作用效果。方法 SD大鼠70只用热液烧伤的方法(75℃、8 s)制成背部深II度烧伤创面,烧伤大鼠随机分为实验组和对照组(每组35只)。对照组( C组):创面局部外用不含rhGM-CSF的凝胶基质,实验组( E组):创面局部外用rhGM-CSF凝胶(100μg/10 g)。分别于制创后1、3、5、7、10、14和21 d观察2组动物创面情况并摄像,记录创面溶痂时间;用图像分析软件计算不同时相点的溶痂率;并于不同的时相点取创面组织, HE染色观察创面组织形态及修复情况。结果从烧伤后第5天起各时相点,实验组大鼠创面溶痂率与对照组表现出差异;实验组创面溶痂时间为(10.73±2.47)d较对照组(14.26±2.65)d显著缩短(P<0.01);实验组和对照组创面愈合时间分别为(16.21±1.27)d和(18.05±1.36)d,差异有非常显著性(P<0.01)。结论外源性rhGM-CSF的应用可促进深II度烧伤创面溶痂,从而促进深II度烧伤创面愈合。%Objective To explore the effect of recombinant human granulocyte-macrophage colony-stimulating factor ( rh-GM-CSF) gel on crust melting and wound healing of deep partial thickness scalding SD rats .Methods A total of 70 SD rats were scalded on the back using the method of thermal-water burns at 75℃for 8s.A deep partial-thickness burn was created in all the rats.The scalded rats were randomly divided into two groups (35 for each group).The control group(C) was treated with gel matrix while the experiment group ( E) was treated with rhGM-CSF gel .The wounds treatment of the two groups lasted 21 days.Observations of the wounds were made by photograph at various time-points.In addition, the removal time of wound eschar in all the rats was recorded .The percentages of removal area in the

  9. Effect of boschniakia rossica on expression of GST-P, p53 and p21ras proteinsin early-stage chemical hepatocarcinogenesis and its anti-inflammatoryactivities in rats

    Institute of Scientific and Technical Information of China (English)

    Zong Zhu Yin; Hai Ling Jin; Xue Zhe Yin; Tian Zhu Li; Ji Shu Quan; Zeng Nan Jin

    2000-01-01

    AIM To investigate the effect of boschniakia rossica (BR) extract on expression of GST-P, p53 and p21fasproteins in early-stage chemical hepatocarcinogenesis in rats and its anti-inflammatory actions.METHODS The expression of tumor marker, placental form glutathione S-transferase (GST-P), p53 and p21ras proteins were investigated by immunohistochemical techniques and ABC method. Anti-inflammatoryactivities of BR were observed by xylene and croton oil-induced mouse ear edema, carrageenin, histamineand hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet-induced mousegranuloma formation methods.RESULTS The 500 mg/kg of BR-H2O extract fractionated from BR-Methanol extract had inhibitory effecton the formation of DEN-induced GST-P-positive foci in rat liver and the expression of mutant p53 and p21fasprotein was lower than that of hepatic preneoplastic lesions. Both CH2Cl2 and H2O extract from BR haveinhibitory effect in xylene and croton oil-induced mouse ear edema. BR-H2O extract exhibited inhibitoryeffect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in ratsand cotton pellet-induced granuloma formation in mice.CONCLUSION BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in earlystage of rat chemical hepatocarcinogenesis. Both CH2Cl2 and H2O extract from BR exerted anti-inflammatory effect in rats and mice.

  10. The effect of 2,4-diamino-6-hydroxy-pyrimidine on postburn Staphylococcus aureus sepsis in rats

    Institute of Scientific and Technical Information of China (English)

    Li Hongyun; Yao Yongming; Shi ZhiGuo; Dong Ning; Yu Yan; Lu Lianrong; Sheng Zhiyong

    2002-01-01

    treatment, the 6-hour mortality was 55.6% (20/36), while it was only 25.0% in DAHP-treated animals (4/16, P=0. 08). Conclusions: Early treatment with DAHP might be a potential strategy to prevent the development of postburn Staphylococcal sepsis, which appears to be associated with down-regulation of biopterin and NO formation by DAHP.

  11. Detection of target staphylococcal enterotoxin B antigen in orange juice and popular carbonated beverages using antibody-dependent antigen-capture assays.

    Science.gov (United States)

    Principato, MaryAnn; Njoroge, Joyce M; Perlloni, Andrei; O' Donnell, Michael; Boyle, Thomas; Jones, Robert L

    2010-10-01

    There is a critical need for qualitative and quantitative methodologies that provide the rapid and accurate detection of food contaminants in complex food matrices. However, the sensitivity of the assay can be affected when antigen-capture is applied to certain foods or beverages that are extremely acidic. This study was undertaken to assess the effects of orange juice and popular carbonated soft drink upon the fidelity of antibody-based antigen-capture assays and to develop simple approaches that could rescue assay performance without the introduction of additional or extensive extraction procedures. We examined the effects of orange juice and a variety of popular carbonated soft drink beverages upon a quantitative Interleukin-2 (IL-2) enzyme-linked immunosorbent assay (ELISA) assay system and a lateral flow device (LFD) adapted for the detection of staphylococcal enterotoxin B (SEB) in foods. Alterations in the performance and sensitivity of the assay were directly attributable to the food matrix, and alterations in pH were especially critical. The results demonstrate that approaches such as an alteration of pH and the use of milk as a blocking agent, either singly or in combination, will partially rescue ELISA performance. The same approaches permit lateral flow to efficiently detect antigen. Practical Application: The authors present ways to rescue an ELISA assay compromised by acidity in beverages and show that either the alteration of pH, or the use of milk as a blocking agent are not always capable of restoring the assay to its intended efficiency. However, the same methods, when employed with lateral flow technology, are rapid and extremely successful.

  12. Characterization of the staphylococcal cassette chromosome composite island of Staphylococcus haemolyticus SH32, a methicillin-resistant clinical isolate from China.

    Directory of Open Access Journals (Sweden)

    Dongliang Yu

    Full Text Available Staphylococcal cassette chromosome (SCC elements contribute considerably to virulence and resistance to antibiotic agents in staphylococci. SCC elements in coagulase-negative staphylococci (CoNS are highly diverse and there is evidence suggesting that they serve as a reservoir for antibiotic resistance genes in methicillin-resistant Staphylococcus aureus (MRSA. However, only a small number of SCC elements have been characterized in CoNS and their exact roles in the emergence and evolution of MRSA remain to be demonstrated. Here, we determined the structure of an SCC composite island (CISH32 found in the clinical Staphylococcus haemolyticus isolate SH32 by whole-genome DNA sequencing. CISH32 was 48 kb in length and mainly composed of two imperfect SCC elements, namely (i a ΨSCCmec(SH32 part containing a class C1 mec gene complex but lacking ccr genes and (ii a SCCSH32 part with a ccrA5B3 gene complex but lacking mec genes. In addition, CISH32 contained a type III restriction-modification system and several resistance loci, for example genes conferring resistance to cadmium and arsenic. ΨSCCmec(SH32 is almost entirely identical to a pseudo SCCmec element found in S. haemolyticus WCH1 and shares pronounced sequence similarity to a ΨSCCmec element of S. haemolyticus JCSC1435. However, staphylococci other than S. haemolyticus, including S. aureus and S. epidermidis, contain homologs of SCCSH32 that are more similar to SCCSH32 than those elements found in S. haemolyticus, suggesting that CISH32 of S. haemolyticus SH32 was assembled in recent evolutionary events. Moreover, the composite structure of CISH32 indicates that the detection of class C1 mec and ccrA5B3 gene complexes in S. haemolyticus does not always indicate the existence of a UT9-type SCCmec element, which has remained questionable.

  13. Efficacy of the Quorum Sensing Inhibitor FS10 Alone and in Combination with Tigecycline in an Animal Model of Staphylococcal Infected Wound.

    Directory of Open Access Journals (Sweden)

    Oriana Simonetti

    Full Text Available In staphylococci, quorum sensing regulates both biofilm formation and toxin production, moreover it has been demonstrated to be inhibited by RNAIII inhibiting peptide (RIP. Aim our study was to evaluate the in vitro activity and its in vivo efficacy of the combined administration of FS10, a novel RIP derivative, and tigecycline in an animal model of methicillin-resistant (MR and methicillin-sensitive (MS Staphylococcus aureus wound infection. Using a 1.x2 cm template, one full thickness wound was established through the panniculus carnosus on the back subcutaneous tissue of each animal. Infection was determined by inoculation of 5x107 CFU/ml of bacteria, that produced an abscess within 24 h, after this, treatment was initiated. The study included, for each strain, a control group without infection, a control infected group that did not receive any treatment and a control infected group with drug-free foam dressing, and three infected groups treated, respectively, with: FS10-soaked foam dressing (containing 20 μg FS10, daily intraperitoneal tigecycline (7 mg/Kg, FS10-soaked foam dressing (containing 20 μg FS10 and daily intraperitoneal injections of tigecycline (7 mg/Kg. The main outcome measures were quantitative culture and histological examination of tissue repair. The highest inhibition of infection was achieved in the group that received FS10-soaked and parenteral tigecycline reducing the bacterial load from 107 CFU/ml to about 103 CFU/g for MSSA and to about 104 CFU/g for MRSA. The group treated with FS10-soaked foam dressing associated with parenteral tigecycline showed, histologically, better overall healing with epithelialization and collagen scores significantly higher than those of the other groups in both strains. In conclusion, the combined use of topical FS10 with i.p. tigecycline induced positive interaction in vivo, resulting in an enhanced therapeutic benefit versus staphylococcal infections in murine wound models.

  14. Effect of different ionizing radiation doses and dose rates, using Cobalt-60 and electrons beam sources, on the staphylococcal enterotoxin inoculated in mechanically deboned chicken meat

    Energy Technology Data Exchange (ETDEWEB)

    Pomarico Neto, Walter; Brito, Poliana de Paula; Azevedo, Heliana de; Roque, Claudio Vitor; Fukuma, Henrique Takuji, E-mail: pbrito@cnen.gov.br, E-mail: hazevedo@cnen.gov.br, E-mail: cvroque@cnen.gov.br, E-mail: htfukuma@cnen.gov.br [Brazilian Nuclear Energy Commission (LAPOC/CNEN), Pocos de Caldas, MG (Brazil); Kodama, Yasko, E-mail: ykodama@ipen.br [Nuclear and Energy Research Institute (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Miya, Norma Terugo Nago; Pereira, Jose Luiz, E-mail: miya@fea.unicamp.br, E-mail: pereira@fea.unicamp.br [Campinas State University (UNICAMP), SP (Brazil). Dept. of Food Sciences

    2011-07-01

    The purpose of food irradiation is the destruction of present pathogenic microorganisms and the increase of shelf life of foods. To achieve this process, the source of cobalt-60 and the electron accelerator can be used. The mechanically deboned chicken meat (MDCM) is used for the production of traditional meat products, and it may come to present pathogenic microorganisms such as staphylococcus aureus, a bacterium that produces enterotoxin, which causes food poisoning. The objective of this study is to analyze the effect of ionizing irradiation with different doses and dose rates, deriving from different radiation sources, on staphylococcal enterotoxin type B (SEB) in the MDCM. 50 g samples of MDCM were prepared in a batch of 6 kg of MDCM. The samples were contaminated, with the exception of the control, with SEB in amounts of about 100 ng. Then they were conditioned in a transparent bag made of low density polyethylene, frozen at -18{+-}1 deg C overnight and irradiated in these conditions with doses of 0.0 kGy (control), 1.5 kGy and 3.0 kGy, and with three different dose rates, both in the Cobalt-60 and the electron accelerator. The experiments were conducted in quintuplicate. The SEB extraction from the MDCM was performed according to the protocol recommended by the manufacturer of the kit VIDAS Staph Enterotoxin II (bioMerrieux). The principle of mass balance was used to determine the actual amount of SEB removed by irradiation. The treatment that presented the best results was the one with a dose of 1.5 kGy, high dose rate of the electron accelerator. (author)

  15. Therapeutic inhibition of pro-inflammatory signaling and toxicity to staphylococcal enterotoxin B by a synthetic dimeric BB-loop mimetic of MyD88.

    Directory of Open Access Journals (Sweden)

    Teri L Kissner

    Full Text Available Staphylococcal enterotoxin B (SEB exposure triggers an exaggerated pro-inflammatory cytokine response that often leads to toxic shock syndrome (TSS associated with organ failure and death. MyD88 mediates pro-inflammatory cytokine signaling induced by SEB exposure and MyD88(-/- mice are resistant to SEB intoxication, suggesting that MyD88 may be a potential target for therapeutic intervention. We targeted the BB loop region of the Toll/IL-1 receptor (TIR domain of MyD88 to develop small-molecule therapeutics. Here, we report that a synthetic compound (EM-163, mimic to dimeric form of BB-loop of MyD88 attenuated tumor necrosis factor (TNF- α, interferon (IFN-γ, interleukin (IL-1β, IL-2 and IL-6 production in human primary cells, whether administered pre- or post-SEB exposure. Results from a direct binding assay, and from MyD88 co-transfection/co-immunoprecipitation experiments, suggest that EM-163 inhibits TIR-TIR domain interaction. Additional results indicate that EM-163 prevents MyD88 from mediating downstream signaling. In an NF-kB-driven reporter assay of lipopolysaccharide-stimulated MyD88 signaling, EM-163 demonstrated a dose-dependent inhibition of reporter activity as well as TNF-α and IL-1β production. Importantly, administration of EM-163 pre- or post exposure to a lethal dose of SEB abrogated pro-inflammatory cytokine responses and protected mice from toxic shock-induced death. Taken together, our results suggest that EM-163 exhibits a potential for therapeutic use against SEB intoxication.

  16. Emergence of sequence type 779 methicillin-resistant Staphylococcus aureus harboring a novel pseudo staphylococcal cassette chromosome mec (SCCmec)-SCC-SCCCRISPR composite element in Irish hospitals.

    LENUS (Irish Health Repository)

    Kinnevey, Peter M

    2013-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) has been a major cause of nosocomial infection in Irish hospitals for 4 decades, and replacement of predominant MRSA clones has occurred several times. An MRSA isolate recovered in 2006 as part of a larger study of sporadic MRSA exhibited a rare spa (t878) and multilocus sequence (ST779) type and was nontypeable by PCR- and DNA microarray-based staphylococcal cassette chromosome mec (SCCmec) element typing. Whole-genome sequencing revealed the presence of a novel 51-kb composite island (CI) element with three distinct domains, each flanked by direct repeat and inverted repeat sequences, including (i) a pseudo SCCmec element (16.3 kb) carrying mecA with a novel mec class region, a fusidic acid resistance gene (fusC), and two copper resistance genes (copB and copC) but lacking ccr genes; (ii) an SCC element (17.5 kb) carrying a novel ccrAB4 allele; and (iii) an SCC element (17.4 kb) carrying a novel ccrC allele and a clustered regularly interspaced short palindromic repeat (CRISPR) region. The novel CI was subsequently identified by PCR in an additional 13 t878\\/ST779 MRSA isolates, six from bloodstream infections, recovered between 2006 and 2011 in 11 hospitals. Analysis of open reading frames (ORFs) carried by the CI showed amino acid sequence similarity of 44 to 100% to ORFs from S. aureus and coagulase-negative staphylococci (CoNS). These findings provide further evidence of genetic transfer between S. aureus and CoNS and show how this contributes to the emergence of novel SCCmec elements and MRSA strains. Ongoing surveillance of this MRSA strain is warranted and will require updating of currently used SCCmec typing methods.

  17. Efficacy of the Quorum Sensing Inhibitor FS10 Alone and in Combination with Tigecycline in an Animal Model of Staphylococcal Infected Wound.

    Science.gov (United States)

    Simonetti, Oriana; Cirioni, Oscar; Cacciatore, Ivana; Baldassarre, Leonardo; Orlando, Fiorenza; Pierpaoli, Elisa; Lucarini, Guendalina; Orsetti, Elena; Provinciali, Mauro; Fornasari, Erika; Di Stefano, Antonio; Giacometti, Andrea; Offidani, Annamaria

    2016-01-01

    In staphylococci, quorum sensing regulates both biofilm formation and toxin production, moreover it has been demonstrated to be inhibited by RNAIII inhibiting peptide (RIP). Aim our study was to evaluate the in vitro activity and its in vivo efficacy of the combined administration of FS10, a novel RIP derivative, and tigecycline in an animal model of methicillin-resistant (MR) and methicillin-sensitive (MS) Staphylococcus aureus wound infection. Using a 1.x2 cm template, one full thickness wound was established through the panniculus carnosus on the back subcutaneous tissue of each animal. Infection was determined by inoculation of 5x107 CFU/ml of bacteria, that produced an abscess within 24 h, after this, treatment was initiated. The study included, for each strain, a control group without infection, a control infected group that did not receive any treatment and a control infected group with drug-free foam dressing, and three infected groups treated, respectively, with: FS10-soaked foam dressing (containing 20 μg FS10), daily intraperitoneal tigecycline (7 mg/Kg), FS10-soaked foam dressing (containing 20 μg FS10) and daily intraperitoneal injections of tigecycline (7 mg/Kg). The main outcome measures were quantitative culture and histological examination of tissue repair. The highest inhibition of infection was achieved in the group that received FS10-soaked and parenteral tigecycline reducing the bacterial load from 107 CFU/ml to about 103 CFU/g for MSSA and to about 104 CFU/g for MRSA. The group treated with FS10-soaked foam dressing associated with parenteral tigecycline showed, histologically, better overall healing with epithelialization and collagen scores significantly higher than those of the other groups in both strains. In conclusion, the combined use of topical FS10 with i.p. tigecycline induced positive interaction in vivo, resulting in an enhanced therapeutic benefit versus staphylococcal infections in murine wound models.

  18. A combined multi-virulence-locus sequence typing and Staphylococcal Cassette Chromosome mec typing scheme possesses enhanced discriminatory power for genotyping MRSA.

    Science.gov (United States)

    Verghese, Bindhu; Schwalm, Nathan D; Dudley, Edward G; Knabel, Stephen J

    2012-12-01

    Methicillin-resistant Staphylococcus aureus (MRSA) remains a major threat to human populations worldwide. Knowing the extent of MRSA genetic diversity within a healthcare facility may provide important insights into the epidemiology of this important pathogen. MRSA isolates recovered from nasal swabs of patients entering the Intensive Care Unit of the Penn State Milton S. Hershey Medical Center, USA, from 2008 to 2009 were genotyped using Staphylococcal Cassette Chromosome mec (SCCmec), multilocus sequence typing (MLST) and a newly developed multi-virulence-locus sequence typing (MVLST) scheme. Sequence data for seven housekeeping genes (arcC, aroE, glpF, gmk, pta, tpi and yqiL) and six virulence genes (alt, essC, geh, hlgA, htrA and sdrC) were used for MLST and MVLST analyses, respectively. MLST identified 12 sequence types (STs) within the hospital isolates. One ST designated ST5 was the most common subtype (38.8%) followed by ST105 (22.4%) and ST8 (16.4%). In contrast, MVLST identified 29 STs (Virulence Types, VTs) from the same set of isolates, with VT6 (32.8%) being the predominant subtype followed by VT9 (8.9%) and VT2 (8.9%). Subsequent analysis of 25 MRSA isolates associated with an outbreak at a Pennsylvania state prison revealed all isolates were VT2 and SCCmec type IVa. These results suggest that a combination of MVLST and SCCmec typing may clarify the epidemiology of MRSA. Additional research with a more diverse set of strains and correlation with conventional epidemiologic data are needed to validate this new subtyping strategy. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Production and purification of staphylococcal nuclease in Lactococcus lactis using a new expression-secretion system and a pH-regulated mini-reactor

    Directory of Open Access Journals (Sweden)

    Duvignau Thomas

    2010-05-01

    Full Text Available Abstract Background Staphylococcal (or micrococcal nuclease or thermonuclease (SNase or Nuc is a naturally-secreted nucleic acid degrading enzyme that participates in Staphylococcus aureus spread in the infected host. Purified Nuc protein can be used as an exogenous reagent to clear cellular extracts and improve protein purification. Here, a recombinant form of Nuc was produced and secreted in a Gram-positive host, Lactococcus lactis, and purified from the culture medium. Results The gene segment corresponding to the S. aureus nuclease without its signal peptide was cloned in an expression-secretion vector. It was then fused to a lactococcal sequence encoding a signal peptide, and expressed under the control of a lactococcal promoter that is inducible by zinc starvation. An L. lactis subsp cremoris model strain (MG1363 transformed with the resulting plasmid was grown in either of two media (GM17v and CDM that are free of animal compounds, allowing GMP (Good Manufacturing Practice production. Induction conditions (concentration of the metal chelator EDTA and timing of addition in small-scale pH-regulated fermentors were optimized using LacMF (Lactis Multi-Fermentor, a home-made parallel fermentation control system able to monitor 12 reactors simultaneously. Large amounts of recombinant Nuc (rNuc were produced and secreted in both media, and rNuc was purified from GM17v medium in a single-step procedure. Conclusions In L. lactis, rNuc production and secretion were optimal after induction by 0.5 mM EDTA in small scale (200 mL GM17v exponential phase cultures (at an OD600 of 2, leading to a maximal protein yield of 210 mg per L of culture medium. Purified rNuc was highly active, displaying a specific activity of 2000 U/mg.

  20. Molecular Basis for Complement Recognition and Inhibition Determined by Crystallographic Studies of the Staphylococcal Complement Inhibitor (SCIN) Bound to C3c and C3b

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, Brandon L.; Ramyar, Kasra X.; Tzekou, Apostolia; Ricklin, Daniel; McWhorter, William J.; Lambris, John D.; Geisbrecht, Brian V. (UMKC); (UPENN)

    2010-10-22

    The human complement system plays an essential role in innate and adaptive immunity by marking and eliminating microbial intruders. Activation of complement on foreign surfaces results in proteolytic cleavage of complement component 3 (C3) into the potent opsonin C3b, which triggers a variety of immune responses and participates in a self-amplification loop mediated by a multi-protein assembly known as the C3 convertase. The human pathogen Staphylococcus aureus has evolved a sophisticated and potent complement evasion strategy, which is predicated upon an arsenal of potent inhibitory proteins. One of these, the staphylococcal complement inhibitor (SCIN), acts at the level of the C3 convertase (C3bBb) and impairs downstream complement function by trapping the convertase in a stable but inactive state. Previously, we have shown that SCIN binds C3b directly and competitively inhibits binding of human factor H and, to a lesser degree, that of factor B to C3b. Here, we report the co-crystal structures of SCIN bound to C3b and C3c at 7.5 and 3.5 {angstrom} limiting resolution, respectively, and show that SCIN binds a critical functional area on C3b. Most significantly, the SCIN binding site sterically occludes the binding sites of both factor H and factor B. Our results give insight into SCIN binding to activated derivatives of C3, explain how SCIN can recognize C3b in the absence of other complement components, and provide a structural basis for the competitive C3b-binding properties of SCIN. In the future, this may suggest templates for the design of novel complement inhibitors based upon the SCIN structure.

  1. Research on detection of Staphylococcal enterotoxin A, B, C, E by PCR%PCR技术检测金黄色葡萄球菌肠毒素A、B、C、E基因的研究

    Institute of Scientific and Technical Information of China (English)

    何晖; 刘华章; 魏泉德

    2012-01-01

    Objective:To found a method for detection of Staphylococcal enterotoxin SEA,SEB,SEC and SEE all on the same polymerase chain reaction (PCR) instrument in the same reaction condition by PCR, and diagnose the food poisoning by Staphylococcal enterotoxin. Methods; According to the published gene sequence encoding of SEA,SEB, SEC and SEE, 4 pairs of primers were designed for specific amplification. Through the optimization of reaction conditions, a method was established to analyze the specificity and sensitivity, and test the PCR products' sequence. Results; The method had good specificity and sensitivity,it can distinguish different genes above, the sensitivity was 0.8-1.0 x 102 CFU/ml in this research. Conclusion: This method can detect Staphylococcal enterotoxin SEA, SEB, SEC and SEE all on the same PCR instrument in the same reaction condition, and distinguish die different types of the Staphylococcal enterotoxin.%目的:建立检测金黄色葡萄球菌肠毒素SEA 、SEB、SEC、SEE的PCR方法,实现同一台PCR仪、同一PCR反应条件下同时检测SEA、SEB、SEC 、SEE,用于金黄色葡萄球菌肠毒素食物中毒的诊断.方法:根据已公布的SEA、SEB、SEC、SEE编码的基因序列,设计4对引物,优化反应条件,建立检测方法,对特异性和敏感性进行分析,并对扩增产物进行测序鉴定.结果:方法特异性分析表明能很好区分各种肠毒素基因型,灵敏度达0.8 - 1.0 × 102 CFU/ml.结论:建市了一种在同一台PCR仪、同一PCR条件下同时检测SEA、SEB、SEC、SEE的方法,可用于金黄色葡萄球菌肠毒素食物中毒的诊断和肠毒素的分型检测.

  2. Effects of staphylococcal enterotoxin B or enterotoxin C on the proliferation and activation of T lymphocytes%超抗原SEB或SEC对T细胞增殖和活化的影响

    Institute of Scientific and Technical Information of China (English)

    李明; 邢飞跃; 李岩

    2005-01-01

    目的:探讨超抗原金黄色葡萄球菌肠毒素B(staphylococcal enterotoxin B,SEB)和金黄色葡萄球菌肠毒素C(staphylococcal enterotoxin C,SEC)在体外对T细胞增殖和活化的影响.方法:不同浓度SEB或SEC在体外刺激T细胞,采用MTT法检测T细胞增殖水平,在流式细胞仪(flow cytometer,FCM)上检测T细胞受不同浓度SEB或SEC刺激后不同时间其早期活化标志CD69的表达水平.结果:SEB或SEC在体外能明显激活T细胞,以浓度为100μg/L时活化T细胞的作用最强,刺激时间以4~8 h为佳,而对T细胞的增殖无明显影响.SEB和SEC活化T细胞的作用强度之间无明显差异.结论:以上结果提示,SEB或SEC具有很强的刺激T细胞活化的能力,为其应用于肿瘤免疫治疗提供了一定依据.

  3. Role of the pre-neck appendage protein (Dpo7 from phage vB_SepiS-phiIPLA7 as an anti-biofilm agent in staphylococcal species

    Directory of Open Access Journals (Sweden)

    Diana eGutiérrez

    2015-11-01

    Full Text Available Staphylococcus epidermidis and Staphylococcus aureus are important causative agents of hospital-acquired infections and bacteremia, likely due to their ability to form biofilms. The production of a dense exopolysaccharide (EPS matrix enclosing the cells slows the penetration of antibiotic down, resulting in therapy failure. The exopolysaccharide depolymerase (Dpo7 derived from bacteriophage vB_SepiS-phiIPLA7, was overexpressed in E. coli and characterized. A dose dependent but time independent response was observed after treatment of staphylococcal 24 h-biofilms with Dpo7. Maximum removal (>90% of biofilm-attached cells was obtained with 0.15 µM of Dpo7 in all polysaccharide producer strains but Dpo7 failed to eliminate polysaccharide-independent biofilm formed by S. aureus V329. Moreover, the pre-treatment of polystyrene surfaces with Dpo7 reduced the biofilm biomass by 53-85% in the 67% of the tested strains. This study supports the use of phage-encoded exopolysaccharide depolymerases to prevent and disperse staphylococcal biofilms, thereby making bacteria more susceptible to the action of antimicrobials.

  4. Natural indoles, indole-3-carbinol and 3,3′-diindolymethane, inhibit T cell activation by staphylococcal enterotoxin B through epigenetic regulation involving HDAC expression

    Energy Technology Data Exchange (ETDEWEB)

    Busbee, Philip B.; Nagarkatti, Mitzi; Nagarkatti, Prakash S., E-mail: prakash@mailbox.sc.edu

    2014-01-01

    Staphylococcal enterotoxin B (SEB) is a potent exotoxin produced by the Staphylococcus aureus. This toxin is classified as a superantigen because of its ability to directly bind with MHC-II class molecules followed by activation of a large proportion of T cells bearing specific Vβ-T cell receptors. Commonly associated with classic food poisoning, SEB has also been shown to induce toxic shock syndrome, and is also considered to be a potential biological warfare agent because it is easily aerosolized. In the present study, we assessed the ability of indole-3-carbinol (I3C) and one of its byproducts, 3,3′-diindolylmethane (DIM), found in cruciferous vegetables, to counteract the effects of SEB-induced activation of T cells in mice. Both I3C and DIM were found to decrease the activation, proliferation, and cytokine production by SEB-activated Vβ8{sup +} T cells in vitro and in vivo. Interestingly, inhibitors of histone deacetylase class I (HDAC-I), but not class II (HDAC-II), showed significant decrease in SEB-induced T cell activation and cytokine production, thereby suggesting that epigenetic modulation plays a critical role in the regulation of SEB-induced inflammation. In addition, I3C and DIM caused a decrease in HDAC-I but not HDAC-II in SEB-activated T cells, thereby suggesting that I3C and DIM may inhibit SEB-mediated T cell activation by acting as HDAC-I inhibitors. These studies not only suggest for the first time that plant-derived indoles are potent suppressors of SEB-induced T cell activation and cytokine storm but also that they may mediate these effects by acting as HDAC inhibitors. - Highlights: • I3C and DIM reduce SEB-induced T cell activation and inflammatory cytokines. • Inhibiting class I HDACs reduces T cell activation and inflammatory cytokines. • Inhibiting class II HDACs increases T cell activation and inflammatory cytokines. • I3C and DIM selectively reduce mRNA expression of class I HDACs. • Novel use and mechanism to counteract

  5. Preparation and activity of conjugate of monoclonal antibody HAbl8 against hepatoma F(ab')2 fragment and staphylococcal enterotoxin A

    Institute of Scientific and Technical Information of China (English)

    Lian Jun Yang; Yan Fang Sui; Zhi Nan Chen

    2001-01-01

    AIM To prepare the conjugate of staphylococcal enterotoxin A (SEA) protein which is a bacterial SAg and the F(ab')2 fragment of mAb HAbl8 against human hepatocellular carcinoma (HCC), and identify its activity in order to use SAg in the targeting therapy of HCC.METHODS MAb HAbl8 was extracted from the abdominal dropsy of Balb/ c mice, and was purified through chromatography column SP-40HR with Fast protein liquid chromatography (FPLC) system. The F(ab')2 fragment of mAb HAb18 was prepared by papainic digestion method. The conjugate of mAb HAb18 F(ab')2fragment and SEA was prepared with chemical conjugating reagent N-succinimidyl-3-( 2-pyridyldithio) propionate (SPDP) and purified through chromatography column Superose 12with FPLC system. The molecular mass and purity of each collected peak were identified with SDS-PAGE assay. The protein content was assayed by Lowry's method. The antibody activity of HAb18 F (ab')2 against HCC in the conjugate was identified by indirect immunocytochemical ABC method, and the activity of SEA in the conjugate to activate peripheral blood mononuclear cells (PBMC) was identified with MTT assay.RESULTS The lgG mAb HAb18 was extracted,and purified successfully. Immunocytochemical staining demonstrated that it reacted with most of HHCC cells of human HCC cell line. There were two peaks in the process of purification of the prepared HAb18 F(ab)2-SEA conjugate. SDS-PAGE assay demonstrated that the molecular mass of the first peak was about 130 ku, and the second peak was the mixture of about 45 ku and a little 100 ku proteins. The immunocytochemical staining was similar in HAb18 F (ab ')2-SEAconjugate and HAb18 F (ab ')2, i.e., thecytoplasm and/or cell membranes of most HHCC cells were positively stained. The MTT assay showed that the optical absorbance (A) value at 490 nm of HAb18 F (ab')2-SEA conjugate was 0.182 ± 0.012, that of negative control was 0.033± 0.009, and there was significant difference between them ( P < 0.05).CONCLUSION

  6. Staphylococcal Entertotoxins of the Enterotoxin Gene Cluster (egcSEs Induce Nitrous Oxide- and Cytokine Dependent Tumor Cell Apoptosis in a Broad Panel of Human Tumor Cells

    Directory of Open Access Journals (Sweden)

    David eTerman

    2013-08-01

    Full Text Available The egcSEs comprise five genetically linked staphylococcal enterotoxins, SEG, SEI, SElM, SElN and SElO and two pseudotoxins which constitute an operon present in up to 80% of Staphylococcus aureus isolates. A preparation containing theses proteins was recently used to treat advanced lung cancer with pleural effusion. We investigated the hypothesis that egcSEs induce nitrous oxide (NO and associated cytokine production and that these agents may be involved in tumoricidal effects against a broad panel of clinically relevant human tumor cells. Preliminary studies showed that egcSEs and SEA activated T cells (range: 11-25% in a concentration dependent manner. Peripheral blood mononuclear cells (PBMCs stimulated with equimolar quantities of egcSEs expressed NO synthase and generated robust levels of nitrite (range: 200-250 µM, a breakdown product of NO; this reaction was inhibited by NG-monomethyl-L-arginine (L-NMMA (0.3 mM, an NO synthase antagonist. Cell free supernatants (CSFs of all egcSE-stimulated PBMCs were also equally effective in inducing concentration dependent tumor cell apoptosis in a broad panel of human tumor cells. The latter effect was due in part to the generation of NO and TNF-α since it was significantly abolished by L-NMMA, anti-TNF-α antibodies respectively and a combination thereof. A hierarchy of tumor cell sensitivity to these CFSs was as follows: lung carcinoma>osteogenic sarcoma>melanoma>breast carcinoma>neuroblastoma. Notably, SEG induced robust activation of NO/TNFα-dependent tumor cell apoptosis comparable to the other egcSEs and SEA despite TNF-α and IFN-γ levels that were 2 and 8 fold lower respectively than the other egcSEs and SEA. Thus, egcSEs produced by S. aureus induce NO synthase and the increased NO formation together with TNF-α appear to contribute to egcSE-mediated apoptosis against a broad panel of human tumor cells.

  7. The role of the Staphylococcal VraTSR regulatory system on vancomycin resistance and vanA operon expression in vancomycin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Qureshi, Nadia K; Yin, Shaohui; Boyle-Vavra, Susan

    2014-01-01

    Vancomycin is often the preferred treatment for invasive methicillin-resistant Staphylococcus aureus (MRSA) infection. With the increase in incidence of MRSA infections, the use of vancomycin has increased and, as feared, isolates of vancomycin-resistant Staphylococcus aureus (VRSA) have emerged. VRSA isolates have acquired the entercoccal vanA operon contained on transposon (Tn) 1546 residing on a conjugal plasmid. VraTSR is a vancomycin and β-lactam-inducible three-component regulatory system encoded on the S. aureus chromosome that modulates the cell-wall stress response to cell-wall acting antibiotics. Mutation in vraTSR has shown to increase susceptibility to β-lactams and vancomycin in clinical VISA strains and in recombinant strain COLVA-200 which expresses a plasmid borne vanA operon. To date, the role of VraTSR in vanA operon expression in VRSA has not been demonstrated. In this study, the vraTSR operon was deleted from the first clinical VRSA strain (VRS1) by transduction with phage harvested from a USA300 vraTSR operon deletion strain. The absence of the vraTSR operon and presence of the vanA operon were confirmed in the transductant (VRS1Δvra) by PCR. Broth MIC determinations, demonstrated that the vancomycin MIC of VRS1Δvra (64 µg/ml) decreased by 16-fold compared with VRS1 (1024 µg/ml). The effect of the vraTSR operon deletion on expression of the van gene cluster (vanA, vanX and vanR) was examined by quantitative RT-PCR using relative quantification. A 2-5-fold decreased expression of the vanA operon genes occured in strain VRS1Δvra at stationary growth phase compared with the parent strain, VRS1. Both vancomycin resistance and vancomycin-induced expression of vanA and vanR were restored by complementation with a plasmid harboring the vraTSR operon. These findings demonstrate that expression in S. aureus of the horizontally acquired enterococcal vanA gene cluster is enhanced by the staphylococcal three-component cell wall stress regulatory

  8. Staphylococcal resistance revisited: community-acquired methicillin resistant Staphylococcus aureus--an emerging problem for the management of skin and soft tissue infections.

    Science.gov (United States)

    Eady, E Anne; Cove, Jonathan H

    2003-04-01

    In the community non-localized or deep staphylococcal skin and soft tissue infections are typically managed with beta-lactamase stable penicillins. The aims of this review are (1) to evaluate the evidence for the emergence of new strains of community-acquired methicillin resistant Staphylococcus aureus (MRSA), (2) to identify the reasons for their significant association with cutaneous infections, and (3) to consider how they arose and how big a threat they pose to the management of such infections outside hospitals. MRSA are emerging as significant community pathogens, especially in previously healthy children with no recognizable risk factors, and are predominantly associated with skin and soft tissue infections (especially abscesses and cellulitis). When present, risk factors are generally similar to those for infection with methicillin susceptible S. aureus. The MRSA isolates associated with such infections may not be entirely 'new', but could represent the displacement of some hospital clones (e.g. EMRSA-15 or variants thereof) to the community as well as the de-novo generation of novel MRSA clones by multiple horizontal transmissions of the mecA gene into methicillin susceptible S. aureus with different genetic backgrounds, some of which are already circulating globally. Community-acquired MRSA from diverse locations are non multiresistant and almost always contain the novel type IV SCCmec commonly found in coagulase-negative staphylococci, but also in hospital-associated gentamicin susceptible MRSA from France, the paediatric clone and in EMRSA-15. More local data on CA-MRSA infections are needed so that dermatologists and community physicians can assess the risk of such infections amongst their patients and avoid the inappropriate administration of beta-lactams. No simple change in prescribing practices will entirely alleviate selective pressure for the spread of community-acquired MRSA and not exacerbate resistance in pyogenic streptococci, commonly found

  9. Comparative Genotypes, Staphylococcal Cassette Chromosome mec (SCCmec Genes and Antimicrobial Resistance amongst Staphylococcus epidermidis and Staphylococcus haemolyticus Isolates from Infections in Humans and Companion Animals.

    Directory of Open Access Journals (Sweden)

    Brenda A McManus

    staphylococcal species-specific characteristics were identified in relation to antimicrobial resistance genes and phenotypes, SCCmec and ACME.

  10. Early Astronomy

    Science.gov (United States)

    Thurston, Hugh

    The earliest investigations that can be called scientific are concerned with the sky: they are the beginnings of astronomy. Many early civilizations produced astronomical texts, and several cultures that left no written records left monuments and artifacts-ranging from rock paintings to Stonehenge-that show a clear interest in astronomy. Civilizations in China, Mesopotamia, India and Greece had highly developed astronomies, and the astronomy of the Mayas was by no means negligible. Greek astronomy, as developed by the medieval Arab philosophers, evolved into the astronomy of Copernicus. This displaced the earth from the central stationary position that almost all earlier astronomies had assumed. Soon thereafter, in the first decades of the seventeenth century, Kepler found the true shape of the planetary orbits and Galileo introduced the telescope for astronomical observations.

  11. 应用负压治疗技术修复小儿浅Ⅱ度烫伤创面的效果观察%Efficacy observation on application of negative pressure therapy in the treatment of superficial partial-thickness scald wound in children

    Institute of Scientific and Technical Information of China (English)

    申传安; 柴家科; 庹晓晔; 蔡建华; 李东杰; 张琳; 朱华; 蔡金东

    2013-01-01

    目的 观察应用负压治疗技术修复小儿浅Ⅱ度烫伤创面的效果. 方法 将2009年8月-2012年5月笔者单位收治的307例浅Ⅱ度烫伤患儿按照随机数字表法进行分组,负压治疗组145例、对照组162例.负压治疗组患儿于伤后72 h内进行负压治疗(压力为-16 kPa),伤后第9天结束;对照组患儿采用传统的包扎换药方式治疗.对比2组患儿治疗过程中的体温变化、创面修复情况及换药次数;对比负压治疗组治疗前后的创面细菌培养情况,记录负压治疗组治疗后1、2、3d每百分之一烫伤面积的引流液量.数据比较行t检验或x2检验. 结果 负压治疗组患儿出现发热的比例为26.9% (39/145),显著低于对照组[63.6% (103/162),x2 =41.419,P<0.01].伤后第9天,负压治疗组138例患儿的创面完全上皮化愈合,7例零星残余创面经碘伏消毒换药后2d愈合.负压治疗组创面愈合时间为(9.2±0.6)d,明显短于对照组[(10.1±1.6)d,t=6.895,P<0.01].负压治疗组患儿的换药次数为(2.05±0.22)次,显著少于对照组[(4.82±0.81)次,t=39.878,P<0.01].负压治疗组治疗前,17例创面分泌物细菌培养结果呈阳性;治疗后,7例未愈创面分泌物细菌培养结果均为阴性.负压治疗组引流液量与创面大小成正比,治疗后第1、2、3天每百分之一烫伤面积引流液量分别为(9.8±3.2)、(6.2±2.1)、(4.1±1.6)mL. 结论 负压治疗可显著减少换药次数,通过引流创面渗液减轻感染和炎症反应进而促进创面愈合,是治疗小儿浅Ⅱ度烫伤的安全有效方法.%Objective To observe the effect of negative pressure therapy in the treatment of superficial partial-thickness scald in children.Methods Three hundred and seven children with superficial partial-thickness scald hospitalized from August 2009 to May 2012 were divided into negative pressure therapy group (NPT,n =145) and control group (C,n =162) according to the random number table.Patients in

  12. Regulation of Staphylococcal Enterotoxin Biosynthesis

    Science.gov (United States)

    1981-02-05

    enterotoxin B gene (entB) were provided by Dornbusch et al. (25, 38). This group analyzed SEB synthesis in the methicillin resistant (Meer) isolate, S... Dornbusch et al. (25, 38) proposed that entB and mec are linked on a plasmid. Unfortunately, they did not provide any biophysical data to substantiate...to the elucidation of the genetics of resistance. Cohen and Sweeney (39) reported that unlike the findings of Dornbusch et al. (25, 38) the Mecr

  13. Regulation of Staphylococcal Enterotoxin Biosynthesis

    Science.gov (United States)

    1981-05-01

    evtoc’romerPr r pSNI. p)SN2. anmd p)SNt: (Iwo’r 6) PsSl.,0i) shoi uing ’(o (t2.400 a(10 tons). rhvrnoir p~sin (2.5,O(M daltonps.. oral - plasmid D)N...to the expresion of the SEB÷ phenotype. GENETICS OF SEA There have been no reports as yet associating enterotoxin A pro- duction with plasmid DNA, as

  14. Host Adaptation of Staphylococcal Leukocidins

    NARCIS (Netherlands)

    Vrieling, M.

    2016-01-01

    Staphylococcus aureus is a human and animal pathogen of global importance and has the capacity to cause disease in distinct host populations, using a large arsenal of secreted proteins to evade the host immune response. Amongst the immune evasion proteins of S. aureus, secreted cytotoxins play a pre

  15. Staphylococcal response to oxidative stress

    Directory of Open Access Journals (Sweden)

    Rosmarie eGaupp

    2012-03-01

    Full Text Available Staphylococci are a versatile genus of bacteria that are capable of causing acute and chronic infections in diverse host species. The success of staphylococci as pathogens is due in part to their ability to mitigate endogenous and exogenous oxidative and nitrosative stress. Endogenous oxidative stress is a consequence of life in an aerobic environment; whereas, exogenous oxidative and nitrosative stress are often due to the bacteria’s interaction with host immune systems. To overcome the deleterious effects of oxidative and nitrosative stress, staphylococci have evolved protection, detoxification, and repair mechanisms that are controlled by a network of regulators. In this review, we summarize the cellular targets of oxidative stress, the mechanisms by which staphylococci sense oxidative stress and damage, oxidative stress protection and repair mechanisms, and regulation of the oxidative stress response. When possible, special attention is given to how the oxidative stress defense mechanisms help staphylococci control oxidative stress in the host.

  16. 高海拔地区大鼠严重烫伤延迟复苏后肝脏低氧诱导因子-1α和p53的表达与肝脏损伤的关系%Influence of hypoxia inducible factor-1α and p53 expression on liver injury after severe scald with delayed fluid resuscitation in rats in high altitude

    Institute of Scientific and Technical Information of China (English)

    刘毅; 张诚; 刘长海

    2011-01-01

    Objective To explore the influence of hypoxia-inducible factor-1α (HIF-1α) and p53 expression on liver injury after severe scald with delayed resuscitation in high altitude in rats. Methods 240 Wistar rats were employed as the experimental models (TBSA 30%, Ⅲ degree), and they were randomly divided into 3 groups in different altitudes (1 517 m and 3848 m): delayed fluid resuscitation (DFR) group, immediate fluid resuscitation (IFR) group and sham scald (SS) group. The livers of rats were harvested at 1, 6, 12, 24, 72 hours and 168 hours after severe scald respectively. The levels of HIF-1α, p53 and the apoptosis percentage in liver were detected by pathological technique, tissue microarray technique, terminal deoxynucleotidy transferasemediated deoxy-uridine triphosphate nick end labeling, immunohistochemistry and image analysis technique. Results The expression levels of HIF-1α and p53 were much higher in DFR group and IFR group than those in SS group, in 3 848 m altitude and in 1 517 m altitude, and higher in DFR group than those in IFR group (P <0.05). The apoptosis percentage in liver was higher in DFR group than in IFR group, and higher in 3848 m altitude than in 1 517 m altitude (P <0.05). Conclusion HIF-1α expression and activation of p53 apoptotic pathways may play an important role in the hypoxic injury of the liver.%目的 探讨高海拔地区大鼠严重烫伤延迟复苏后肝脏低氧诱导因子-1α(HIF-1α)和p53的表达与肝脏损伤的关系.方法 Wistar大鼠240只,在1 517和3848m两个海拔高度随机分为即时复苏(IFR)组、延迟复苏(DFR)组和假烫伤(SS)组,建立总体表面积30%的Ⅲ°烫伤模型,于伤后1、6、12、24、72和168 h取材.采用组织病理学、组织芯片技术、脱氧核糖核苷酸末端转移酶介导的缺口末端标记(原位末端缺口标记)法、免疫组织化学染色与图像分析技术,观察肝组织病理学改变与细胞凋亡、HIF-1α和p53的表达.结果

  17. Spread of Epidemic MRSA-ST5-IV Clone Encoding PVL as a Major Cause of Community Onset Staphylococcal Infections in Argentinean Children

    Science.gov (United States)

    Sola, Claudia; Egea, Ana L.; Moyano, Alejandro J.; Garnero, Analia; Kevric, Ines; Culasso, Catalina; Vindel, Ana; Lopardo, Horacio; Bocco, José L.

    2012-01-01

    /Significance The dissemination of epidemic MRSA clone, ST5-IV-PVL+ was the main cause of increasing staphylococcal community-onset infections in Argentinean children (2003–2008), conversely to other countries. The predominance of this clone, which has capacity to express the h-VISA phenotype, in healthcare-associated community-onset cases suggests that it has infiltrated into hospital-settings. PMID:22291965

  18. Spread of epidemic MRSA-ST5-IV clone encoding PVL as a major cause of community onset staphylococcal infections in Argentinean children.

    Directory of Open Access Journals (Sweden)

    Claudia Sola

    associated with a SCCmec type closely related to SCCmecIV(2B&5. CONCLUSIONS/SIGNIFICANCE: The dissemination of epidemic MRSA clone, ST5-IV-PVL(+ was the main cause of increasing staphylococcal community-onset infections in Argentinean children (2003-2008, conversely to other countries. The predominance of this clone, which has capacity to express the h-VISA phenotype, in healthcare-associated community-onset cases suggests that it has infiltrated into hospital-settings.

  19. 烧伤小鼠腹腔巨噬细胞肌醇脂质信号系统变化的实验研究%Changes of Inositol Lipid Signal System and Its Effect on the Secretion of TNF of Peritoneal Macrophage Following Severe Scald in Mice

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Purpose:To further study on the mechanism of abnormal macrophages functional changes postburn, insositol lipid signal system and its effects on TNF secretion by peritoneal macrophages (PM() were observed in severely scalded mice. Method: The activity of PL C (phospholipase C), DAG (diacylglycerol), PKC (protein kinase C), and the alternations of IP3 (inositol-1, 4, 5,-triphosphate), Ca2+ and TNF (tumor necrosis factor) in PM( were measured, and the effects of H-7 (1-(5-isoquinolineslfony1)-2-methylpierazine, a specific PKC inhibitor) and W-7 (N-6-amino-hexyl-5-chloro-1-naphthalenesulfonamide, CaM antagonist) on the production of TNF were also observed. Result: It showed that all above-mentioned parameters changes significantly at 6h,12h,24h postburn. There were remarkable decreases of TNF by using H-7. Conclusion: The results suggested that the inositol lipid signal system of PM( is one of the main signal systems participating in the secretion of TNF, and in this system the DAG-PKC signal pathway showed closer relationship than IP3-Ca2+ in the TNF production.%目的:探讨烧伤导致巨噬细胞功能异常的发生机制。方法:测定了严重烧伤小鼠腹腔巨噬细胞(PMΦ)磷酯酶C(PLC)、甘油二酯(DAG)、蛋白激酶C(PKC)的活性,三磷酸肌醇(IP3)、钙离子(Ca2+)、TNF的变化,以及PKC抑制剂H-7和钙调素(CaM)拮抗剂W-7对TNF产生的影响。结果:上述所观察的指标在严重烧伤后6hr、12hr、24hr、都发生了非常明显的变化,PKC抑制剂H-7能够显著地抑制TNF的产生。结论:腹腔巨噬细胞肌醇脂质信号系统是促使TNF分泌的主要信号途径之一,在肌醇脂质信号系统中以DAG-PKC的信号途径与TNF分泌关系更为密切。

  20. 负载超抗原SEB/SEC的树突状细胞体外抗肝癌实验研究%Effects of superantigen staphylococcal enterotoxin B and C-loaded dendritic cells on hepatocarcinoma HcaF

    Institute of Scientific and Technical Information of China (English)

    李明; 李岩; 邢飞跃

    2004-01-01

    目的研究负载金黄色葡萄球菌肠毒素B(staphylococcal enterotoxin B,SEB)和金黄色葡萄球菌肠毒素C(staphylococcal enterotoxin C,SEC)的DC在体外对T细胞的活化能力及其对肝细胞癌HcaF细胞的杀伤作用.方法以SEB/SEC负载DC刺激T细胞,流式细胞仪(flow cytometer,FCM)检测T细胞受刺激后其早期活化标志CD69表达情况和IL-2水平,采用MTT法检测T细胞增殖水平和其对HcaF细胞的杀伤效应.结果SEB/SEC联合DC在体外能明显激活T细胞,以浓度为100 ng/ml时作用最强;SEB/SEC联合DC可以使T细胞活化、增殖并大量分泌IL-2,对HcaF细胞的杀伤率高达83.2±12.8%.以SE作为刺激物负载的DC较以Tag作为刺激物负载的DC有更强的活化T细胞的能力;结论根据以上结果,可以认为SEB/SEC联合DC具有很强的刺激T细胞活化的能力;其活化的T细胞具有很强的杀瘤作用,在肿瘤免疫治疗方面展现出良好的潜在应用前景.

  1. Detection of staphylococcal cassette chromosome mec type XI carrying highly divergent mecA, mecI, mecR1, blaZ, and ccr genes in human clinical isolates of clonal complex 130 methicillin-resistant Staphylococcus aureus.

    LENUS (Irish Health Repository)

    Shore, Anna C

    2011-08-01

    Methicillin resistance in staphylococci is mediated by penicillin binding protein 2a (PBP 2a), encoded by mecA on mobile staphylococcal cassette chromosome mec (SCCmec) elements. In this study, two clonal complex 130 (CC130) methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in Irish hospitals were identified that were phenotypically PBP 2a positive but lacked mecA by conventional PCR and by DNA microarray screening. The isolates were identified as methicillin-susceptible S. aureus using the GeneXpert real-time PCR assay. Whole-genome sequencing of one isolate (M10\\/0061) revealed a 30-kb SCCmec element encoding a class E mec complex with highly divergent blaZ-mecA-mecR1-mecI, a type 8 cassette chromosome recombinase (ccr) complex consisting of ccrA1-ccrB3, an arsenic resistance operon, and flanking direct repeats (DRs). The SCCmec element was almost identical to that of SCCmec type XI (SCCmec XI) identified by the Sanger Institute in sequence type 425 bovine MRSA strain LGA251 listed on the website of the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements. The open reading frames (ORFs) identified within SCCmec XI of M10\\/0061 exhibited 21 to 93% amino acid identity to ORFs in GenBank. A third DR was identified ca. 3 kb downstream of SCCmec XI, indicating the presence of a possible SCC remnant. SCCmec XI was also identified in the second CC130 MRSA isolate by PCR and sequencing. The CC130 MRSA isolates may be of animal origin as previously reported CC130 S. aureus strains were predominantly from bovine sources. The highly divergent nature of SCCmec XI relative to other SCCmec elements indicates that it may have originated in another taxon.

  2. Clinical analysis of staphylococcic epidermidis infections after craniotomy%颅脑术后表皮葡萄球菌性脑膜炎的临床分析

    Institute of Scientific and Technical Information of China (English)

    张伟; 江玉泉; 姜政; 苗立峰; 李俣; 王磊; 张文华

    2008-01-01

    Objectives To investigate the clinical manifestations, risk factors, bacterial resistance to drugs, so the prophylactic measures and therapy for staphylococcic epidermidis meningitis associated with post-neurosurgical patients maybe defined. Methods A retrospective study of the patients, who were performed neurosurgical operations between August 1999 and August 2007 in Qilu Hospital of Shandong University, was made. Staphylococcus epidermidis was cultured from the cerebrospinal fluid of all cases. Results There were 27 positive patients in the series of 6887 operations, containing 16 males and 11 females, age 2-72y, mean age 37.5y. 10 out of 4660 patients for intracranial tumors, 5 out of 787 for craniocerebral trauma, 10 out of 577 for cerebrospinal fluid shunt or external ventrical drainage (2 complicating intracranial tumors), and 4 for other kinds of operations out of 863 patients. The morbidity of infection was 0.21%, 0.64%, 1.73%, and 0.46% respectively, which had significant difference between the third and the others (P<0.05). The general isolating rate of the methicilin resistant staphylococcus epidermidis was 70.4%. All of the cases were sensitive to vancomycin. Conclusions The staphylococcus epidermidis is one of the most important pathogenic bacterias causing postoperative meningitis, which has predilection in patients with cerebrospinal fluid shunt and external ventrical drainage or with contaminated injuries. Though all sensitive to vancomycin, the isolating rate of the methicilin resistant staphylococcus epidermidis is increasing year by year making vancomycin the first choice to cure. Prophylactic antibiotics may be helpful, while rigorous aseptic technique is an important measure to prevent meningitis caused by staphylococcus epidermidis.%目的 分析颅脑术后表皮葡萄球菌性脑膜炎的临床表现、危险因素、细菌耐药性特点,探讨预防措施及治疗方法.方法 回顾性收集1999年8月至2007年8月山东大学齐鲁医

  3. Are We Serving Tea or Scalding Our Children?

    OpenAIRE

    Ilhan Ciftci; Fatih Kara; Kemal Arslan; Zeynep Altunbas; Adnan Abasiyanik

    2013-01-01

    Objective: In this study, we aimed to evaluate preschool children with major burns under inpatient treatment and to examine the reasons for those burns. Method: We retrospectively studied 255 patients between 0 and 5 years of age who suffered from major burns and who received inpatient treatment in the Burn Unit based on the guidelines of the American Burn Association between 2009 and 2011. The patients' age, gender, burn location, cause of burn, time to reach a health center after burn, a...

  4. Fast & Scalded: Capillary Leidenfrost Droplets in micro-Ratches

    CERN Document Server

    Marin, Alvaro G; Römer, Gert-Willem; Lohse, Detlef

    2013-01-01

    In this Fluid Dynamics Videos submitted to the 31st Gallery of Fluid Motion, we illustrate the special dynamics of capillary self-propelled Leidenfrost droplets in micrometr