Hematoma and abscess formation caused by Mycoplasma hominis following cesarean section

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Koshiba, Hisato; Koshiba, Akemi; Daimon, Yasushi; Noguchi, Toshifumi; Iwasaku, Kazuhiro; Kitawaki, Jo

2011-01-01

Mycoplasma species cannot be identified by routine bacteriological culture methods and are resistant to common antimicrobial agents. Mycoplasma hominis usually colonizes the lower urogenital tract and causes pyelonephritis, pelvic inflammatory disease, chorioamnionitis, rupture of fetal membranes, preterm labor, postpartum fever, postabortal fever, and neonatal infection. This organism is highly prevalent in cervicovaginal cultures of sexually active women. M. hominis, M. genitalis, Ureaplasma urealyticum, and U. parvum may invade and infect placental and fetal tissues, leading to adverse pregnancy outcomes. M. hominis occasionally causes nongenitourinary infection of the blood, wounds, central nervous system, joints, or respiratory tract. We present a case of a 27-year-old woman who developed abdominal wound hematoma and abscess after cesarean section. The wound was drained, but her high fever persisted, in spite of antibiotic treatment using flomoxef sodium and imipenem·cilastatin sodium. Because the exudate exhibited M. hominis growth in an anaerobic environment, we administered the quinolone ciprofloxacin. This therapy resolved her fever, and her white blood cell count and C-reactive protein level diminished to the normal ranges. To our knowledge, there are four published articles regarding the isolation of M. hominis from postcesarean incisions. Based on the current study and the literature, infection by this pathogen may cause hematoma formation with or without abscess after cesarean section or in immunosuppressed postoperative patients. In such cases, physicians may need to suspect Mycoplasma infection and initiate appropriate antibacterial treatment as soon as possible in order to avoid persistent fever. PMID:21339933

Meningitis in a Chinese adult patient caused by Mycoplasma hominis: a rare infection and literature review.

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Zhou, Menglan; Wang, Peng; Chen, Sharon; Du, Bin; Du, Jinlong; Wang, Fengdan; Xiao, Meng; Kong, Fanrong; Xu, Yingchun

2016-10-12

Mycoplasma hominis, a well known cause of neonatal infection, has been reported as a pathogen in urogenital infections in adults; however, central nervous system (CNS) infections are rare. We report here the first case of M. hominis meningitis in China, post neurosurgical treatment for an intracerebral haemorrhage in a 71-year-old male. We describe a 71-year-old man who developed M. hominis meningitis after neurosurgical treatment and was successfully treated with combined azithromycin and minocycline therapy of 2 weeks duration, despite delayed treatment because the Gram stain of cerebrospinal fluid (CSF) yielded no visible organisms. The diagnosis required 16S rDNA sequencing analysis of the cultured isolate from CSF. Literature review of M. hominis CNS infections yielded 19 cases (13 instances of brain abscess, 3 of meningitis, 1 spinal cord abscess and 1 subdural empyema each). Delay in diagnosis and initial treatment failure was evident in all cases. With appropriate microbiological testing, antibiotic therapy (ranging from 5 days to 12 weeks) and often, multiple surgical interventions, almost all the patients improved immediately. Both our patient findings and the literature review, highlighted the pathogenic potential of M. hominis together with the challenges prompted by rare infectious diseases in particular for developing countries laboratories with limited diagnostic resources.

Symbiotic Association with Mycoplasma hominis Can Influence Growth Rate, ATP Production, Cytolysis and Inflammatory Response of Trichomonas vaginalis

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Margarita, Valentina; Rappelli, Paola; Dessì, Daniele; Pintus, Gianfranco; Hirt, Robert P.; Fiori, Pier L.

2016-01-01

The symbiosis between the parasitic protist Trichomonas vaginalis and the opportunistic bacterium Mycoplasma hominis is the only one currently described involving two obligate human mucosal symbionts with pathogenic capabilities that can cause independent diseases in the same anatomical site: the lower urogenital tract. Although several aspects of this intriguing microbial partnership have been investigated, many questions on the influence of this symbiosis on the parasite pathobiology still remain unanswered. Here, we examined with in vitro cultures how M. hominis could influence the pathobiology of T. vaginalis by investigating the influence of M. hominis on parasite replication rate, haemolytic activity and ATP production. By comparing isogenic mycoplasma-free T. vaginalis and parasites stably associated with M. hominis we could demonstrate that the latter show a higher replication rate, increased haemolytic activity and are able to produce larger amounts of ATP. In addition, we demonstrated in a T. vaginalis-macrophage co-culture system that M. hominis could modulate an aspect of the innate immuno-response to T. vaginalis infections by influencing the production of nitric oxide (NO) by human macrophages, with the parasite-bacteria symbiosis outcompeting the human cells for the key substrate arginine. These results support a model in which the symbiosis between T. vaginalis and M. hominis influences host-microbes interactions to the benefit of both microbial partners during infections and to the detriment of their host. PMID:27379081

Microbiological and molecular characterization of Staphylococcus hominis isolates from blood.

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Soraya Mendoza-Olazarán

Full Text Available BACKGROUND: Among Coagulase-Negative Staphylococci (CoNS, Staphylococcus hominis represents the third most common organism recoverable from the blood of immunocompromised patients. The aim of this study was to characterize biofilm formation, antibiotic resistance, define the SCCmec (Staphylococcal Chromosomal Cassette mec type, and genetic relatedness of clinical S. hominis isolates. METHODOLOGY: S. hominis blood isolates (n = 21 were screened for biofilm formation using crystal violet staining. Methicillin resistance was evaluated using the cefoxitin disk test and the mecA gene was detected by PCR. Antibiotic resistance was determined by the broth microdilution method. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE and SCCmec typed by multiplex PCR using two different methodologies described for Staphylococcus aureus. RESULTS: Of the S. hominis isolates screened, 47.6% (10/21 were categorized as strong biofilm producers and 23.8% (5/21 as weak producers. Furthermore, 81% (17/21 of the isolates were methicillin resistant and mecA gene carriers. Resistance to ampicillin, erythromycin, and trimethoprim was observed in >70% of isolates screened. Each isolate showed a different PFGE macrorestriction pattern with similarity ranging between 0-95%. Among mecA-positive isolates, 14 (82% harbored a non-typeable SCCmec type: eight isolates were not positive for any ccr complex; four contained the mec complex A ccrAB1 and ccrC, one isolate contained mec complex A, ccrAB4 and ccrC, and one isolate contained the mec complex A, ccrAB1, ccrAB4, and ccrC. Two isolates harbored the association: mec complex A and ccrAB1. Only one strain was typeable as SCCmec III. CONCLUSIONS: The S. hominis isolates analyzed were variable biofilm producers had a high prevalence of methicillin resistance and resistance to other antibiotics, and high genetic diversity. The results of this study strongly suggested that S. hominis isolates harbor

Mycoplasma hominis Induces Mediastinitis after a Tonsillar Abscess

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Anna Grancini

2016-01-01

Full Text Available Mycoplasma hominis is commonly involved in genitourinary tract infections. We report a 59-year-old man who developed a M. hominis-associated mediastinitis following acute tonsillar infection.

Common occurrence of Cryptosporidium hominis in horses and donkeys.

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Jian, Fuchun; Liu, Aiqin; Wang, Rongjun; Zhang, Sumei; Qi, Meng; Zhao, Wei; Shi, Yadong; Wang, Jianling; Wei, Jiujian; Zhang, Longxian; Xiao, Lihua

2016-09-01

Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans. Copyright © 2016 Elsevier B.V. All rights reserved.

Molecular characterization and epidemiological investigation of Cryptosporidium hominis IkA18G1 and C. hominis monkey genotype IiA17, two unusual subtypes diagnosed in Swedish patients.

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Lebbad, Marianne; Winiecka-Krusnell, Jadwiga; Insulander, Mona; Beser, Jessica

2018-05-01

Cryptosporidium hominis is considered a strictly human-adapted species, and it is only occasionally diagnosed in animals. However, two variants, C. hominis monkey genotype and C. hominis Ik, were originally described in non-human hosts, monkeys and horses, respectively. During a Swedish national Cryptosporidium study, where all samples were analyzed at the small subunit rRNA and the 60 kDa (gp60) glycoprotein loci, we identified two patients infected with C. hominis monkey genotype (subtype IiA17) and two infected with C. hominis subtype IkA18G1. The isolates were further analyzed at the actin and the 70 kDa heat shock protein loci, and these analyses showed that these two subtype families are closely related to each other and to human-adapted C. hominis as well as to Cryptosporidium cuniculus. The two patients with C. hominis monkey genotype infection (a father and son) had visited a monkey farm in Thailand prior to infection, while the two cases with C. hominis Ik were unrelated, both probably infected in Sweden. This is the first time that a monkey genotype infection in humans has been related to contact with monkeys and where the gp60 subtype was identified. It is also the first time that human infection caused by C. hominis subtype Ik is described. Even though we were not able to detect any parasites in the animal samples, zoonotic transmission cannot be ruled out in any of these cases because both subtype families are regarded as animal adapted. Copyright © 2018 Elsevier Inc. All rights reserved.

Mycoplasma hominis periaortic abscess following heart-lung transplantation.

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Hagiya, Hideharu; Yoshida, Hisao; Yamamoto, Norihisa; Kimura, Keigo; Ueda, Akiko; Nishi, Isao; Akeda, Yukihiro; Tomono, Kazunori

2017-06-01

We report the first case of Mycoplasma hominis periaortic abscess after heart-lung transplantation. The absence of sternal wound infection delayed the diagnosis, but the patient successfully recovered with debridement surgeries and long-term antibiotic therapy. Owing to the difficulty in detection and the intrinsic resistance to beta-lactams, M. hominis infections are prone to being misdiagnosed and undertreated. M. hominis should be suspected in cases where conventional microbiological identification and treatment approaches fail. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Cardiobacterium hominis and Cardiobacterium valvarum

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Bonavent, Tina Bennett; Nielsen, Xiaohui Chen; Kristensen, Kjeld Skødebjerg

2016-01-01

Introduction: Cardiobacterium hominis and Cardiobacterium valvarum are well known, though rare, etiologic agents of infective endocarditis. Cardiac devices are increasingly implanted. Case Reports: Two cases of infective episodes in pacemaker (PM) treated patients with respectively C. hominis and C...

The Mycoplasma hominis P120 membrane protein contains a 216 amino acid hypervariable domain that is recognized by the human humoral immune response

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Nyvold, Charlotte Guldborg; Birkelund, Svend; Christiansen, Gunna

1997-01-01

In the antigenically heterogeneous species Mycoplasma hominis a monoclonal antibody, mAb 26.7D, was previously found to recognize a 120 kDa polypeptide from M. hominis 7488. This antibody did not react with the type strain PG21. The homologous gene from M. hominis PG21 was cloned and sequenced an...... response. Such a variable domain may be important in evasion of the host's immune response, and thus aid survival of the micro-organism....

Clinical significance and taxonomy of Actinobacillus hominis

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Friis-Møller, Alice; Christensen, J J; Fussing, V

2001-01-01

Clinical findings in 36 immunosuppressed patients with lower respiratory tract infection or bacteremia with Actinobacillus hominis are described. Animal contact was only recorded for three patients; nine patients died despite appropriate antimicrobial treatment. Although infections with this micr......Clinical findings in 36 immunosuppressed patients with lower respiratory tract infection or bacteremia with Actinobacillus hominis are described. Animal contact was only recorded for three patients; nine patients died despite appropriate antimicrobial treatment. Although infections...

Prevalence and antibiotic susceptibility of Mycoplasma hominis and Ureaplasma urealyticum in genital samples collected over 6 years at a Serbian university hospital

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Dusan Skiljevic

2016-01-01

Full Text Available Background: Mycoplasma hominis and Ureaplasma urealyticum are implicated in a wide array of infectious diseases in adults and children. Since some species have innate or acquired resistance to certain types of antibiotics, antibiotic susceptibility testing of mycoplasma isolated from the urogenital tract assumes increasing importance. Aims: To evaluate the prevalence and antibiotic susceptibility of M. hominis and U. urealyticum in genital samples collected between 2007 and 2012. Methods: Three hundred and seventy three patients presenting with symptoms of sexually transmitted diseases, infertility or risky sexual behaviour, who had not taken antibiotics in the previous 6 weeks and had ≥10 WBC per high power field on genital smears were studied. Urethral samples were taken in men and endocervical samples in women. The mycoplasma IST-2 kit was used for organism identification and for testing susceptibility to doxycycline, josamycin, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin and pristinamycin. Results: U. urealyticum was isolated from 42 patients and M. hominis from 11 patients. From 9.8% of isolates, both organisms were grown. All M. hominis isolates were resistant to tetracycline, clarithromycin and erythromycin while U. urealyticum was highly resistant to clarithromycin (94.6%, tetracycline (86.5%, ciprofloxacin (83.8% and erythromycin (83.8%. M. hominis was sensitive to doxycycline (83.3% and ofloxacin (66.7% while most U. urealyticum strains were sensitive to doxycycline (94.6%. Limitations: Inability of the commercial kit used in the study to detect other potentially pathogenic urogenital mycoplasmas (Ureaplasma parvum, Mycoplasma genitalium. Conclusion: There is significant resistance of U. urealyticum and M. hominis to tetracycline and macrolides. The most active tetracycline for genital mycoplasmas was found to be doxycycline, which continues to be the drug of first choice.

Eprinomectin 1% Inyectable in control of Dermatobia hominis in naturally infested cattle

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Cristiano Grisi do Nascimento

2015-12-01

Full Text Available ABSTRACT. Nascimento C.G., Correia T.R., Oliveira G.F., Coumendouros K., Moraes P.A., Calado S.B., Bragaglia G.N., Rosa S.C., Toma S.B. & Scott F.B. [Eprinomectin 1% Inyectable in control of Dermatobia hominis in naturally infested cattle.] Eprinomectina 1%Injetável no controle de Dermatobia hominis em bovinos naturalmente infestados. Revista Brasileira de Medicina Veterinária, 37(Supl.1:81-84, 2015. Programa de Pós-Graduação em Ciências Tecnologia e Inovação em Agropecuária, Universidade Federal Rural do Rio de Janeiro, Campus Seropédica, Ecologia, BR 465 Km 7, Seropédica, RJ 23897-970, Brasil. E-mail: scott.fabio@gmail.com The aim of the study was to evaluate the effectiveness of injectable eprinomectin 1% on the control of Dermatobia hominis in naturally infested cattle. We selected 20 calves, crossbred Gir and Holstein breed, male and female, separated into two groups, control and treated. The control group of animals received no treatment, while the animals of the treated group received eprinomectin formulation of 1% at a single dose of 1 mL/50 kg body weight (200mcg of eprinomectin/kg per injectable route. On days +7 and +14 a count of the total number of live larvae of D. hominis on both sides of the animal for the purpose of evaluation of the effectiveness was performed. Statistical analysis of average living larvae of D. hominis counted among the groups, control and treated, showed that there was a significant difference (p ≤ 0.05 between groups on days +7 and +14. The investigational product showed an efficacy of 100% results in both experimental days. The injectable eprinomectin 1%, shown to be effective in cattle naturally infested by D. hominis (the human bot fly.

Comparative genomic analysis reveals occurrence of genetic recombination in virulent Cryptosporidium hominis subtypes and telomeric gene duplications in Cryptosporidium parvum.

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Guo, Yaqiong; Tang, Kevin; Rowe, Lori A; Li, Na; Roellig, Dawn M; Knipe, Kristine; Frace, Michael; Yang, Chunfu; Feng, Yaoyu; Xiao, Lihua

2015-04-18

Cryptosporidium hominis is a dominant species for human cryptosporidiosis. Within the species, IbA10G2 is the most virulent subtype responsible for all C. hominis-associated outbreaks in Europe and Australia, and is a dominant outbreak subtype in the United States. In recent yearsIaA28R4 is becoming a major new subtype in the United States. In this study, we sequenced the genomes of two field specimens from each of the two subtypes and conducted a comparative genomic analysis of the obtained sequences with those from the only fully sequenced Cryptosporidium parvum genome. Altogether, 8.59-9.05 Mb of Cryptosporidium sequences in 45-767 assembled contigs were obtained from the four specimens, representing 94.36-99.47% coverage of the expected genome. These genomes had complete synteny in gene organization and 96.86-97.0% and 99.72-99.83% nucleotide sequence similarities to the published genomes of C. parvum and C. hominis, respectively. Several major insertions and deletions were seen between C. hominis and C. parvum genomes, involving mostly members of multicopy gene families near telomeres. The four C. hominis genomes were highly similar to each other and divergent from the reference IaA25R3 genome in some highly polymorphic regions. Major sequence differences among the four specimens sequenced in this study were in the 5' and 3' ends of chromosome 6 and the gp60 region, largely the result of genetic recombination. The sequence similarity among specimens of the two dominant outbreak subtypes and genetic recombination in chromosome 6, especially around the putative virulence determinant gp60 region, suggest that genetic recombination plays a potential role in the emergence of hyper-transmissible C. hominis subtypes. The high sequence conservation between C. parvum and C. hominis genomes and significant differences in copy numbers of MEDLE family secreted proteins and insulinase-like proteases indicate that telomeric gene duplications could potentially contribute to

A case of cutaneous myiasis due to Dermatobia hominis in Japan.

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Nagamori, Katsushi; Katayama, Toshiko; Kumagai, Masahiro

2007-08-01

We report the 34th imported case of cutaneous myiasis caused by Dermatobia hominis in Japan, which is not a habitat of the fly. A 41-year-old Japanese man noticed an insect-sting-like papule on his left upper back during his stay in Ecuador in March 2004. After his return home, the lesion gradually increased to become a red subcutaneous nodule with a central pore from which serosanguineous fluid drained. Because antimicrobial treatment under a diagnosis of inflammatory atheroma was ineffective, the lesion was incised and a 3rd instar larva of D. hominis was then found and removed. We checked the literature on D. hominis myiasis reported from Japan, and noted the fact, which nobody had previously pointed out, that in Japan only one case of D. hominis myiasis had been diagnosed correctly before a larva was found, and most of the cases were misdiagnosed and inappropriately treated, including 11 cases given unnecessary resection of the nodules. Doctors in Japan should be aware of myiasis so that patients are neither anxious about the disease nor suffer pain, and doctors avoid performing unnecessary resections of the lesions.

Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum

OpenAIRE

Scott A. Cunningham; Jayawant N. Mandrekar; Jon E. Rosenblatt; Robin Patel

2013-01-01

Objective. We compared laboratory developed real-time PCR assays for detection of Mycoplasma hominis and for detection and differentiation of Ureaplasma urealyticum and parvum to culture using genitourinary specimens submitted for M. hominis and Ureaplasma culture. Methods. 283 genitourinary specimens received in the clinical bacteriology laboratory for M. hominis and Ureaplasma species culture were evaluated. Nucleic acids were extracted using the Total Nucleic Acid Kit on the MagNA Pure 2.0...

Palpebral myiasis in a Danish traveler caused by the human bot-fly (Dermatobia hominis)

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Bangsgaard, Regitze; Holst, Bengt; Krogh, Erik

2000-01-01

ophthalmology, dermatobia hominis, human bot-fly, palpebral myiasis, parasite infection, myiasis......ophthalmology, dermatobia hominis, human bot-fly, palpebral myiasis, parasite infection, myiasis...

Pentatrichomonas hominis: prevalence and molecular characterization in humans, dogs, and monkeys in Northern China.

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Li, Wen-Chao; Ying, Meng; Gong, Peng-Tao; Li, Jian-Hua; Yang, Ju; Li, He; Zhang, Xi-Chen

2016-02-01

Pentatrichomonas hominis is an anaerobic amitochondrial flagellated protist that primarily colonizes the large intestines of a number of species, including cats, dogs, nonhuman primates, and humans. The prevalence of this parasite in dogs, monkeys, and humans is, however, poorly understood. In this study, a total of 362 fecal samples including 252 dogs, 60 monkeys, and 50 humans from northern China were collected for an epidemiological survey of P. hominis infection.The average prevalence of P. hominis infection determined by nested PCR was 27.38% (69/252), 4.00% (2/50), and 46.67% (28/60) in dogs, humans, and monkeys, respectively. The prevalence was significantly higher in 6-month-old dogs (41.53%) and children (7.69%) than in older dogs (14.39%) and adults (0%) (P dogs, monkeys, and humans, especially in children and young dogs. Given the infection prevalence, P. hominis may pose a risk of zoonotic and anthroponotic transmission.

Host Cell Responses to Persistent Mycoplasmas - Different Stages in Infection of HeLa Cells with Mycoplasma hominis

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Hopfe, Miriam; Deenen, René; Degrandi, Daniel; Köhrer, Karl; Henrich, Birgit

2013-01-01

Mycoplasma hominis is a facultative human pathogen primarily associated with bacterial vaginosis and pelvic inflammatory disease, but it is also able to spread to other sites, leading to arthritis or, in neonates, meningitis. With a minimal set of 537 annotated genes, M. hominis is the second smallest self-replicating mycoplasma and thus an ideal model organism for studying the effects of an infectious agent on its host more closely. M. hominis adherence, colonisation and invasion of HeLa cells were characterised in a time-course study using scanning electron microscopy, confocal microscopy and microarray-based analysis of the HeLa cell transcriptome. At 4 h post infection, cytoadherence of M. hominis to the HeLa cell surface was accompanied by differential regulation of 723 host genes (>2 fold change in expression). Genes associated with immune responses and signal transduction pathways were mainly affected and components involved in cell-cycle regulation, growth and death were highly upregulated. At 48 h post infection, when mycoplasma invasion started, 1588 host genes were differentially expressed and expression of genes for lysosome-specific proteins associated with bacterial lysis was detected. In a chronically infected HeLa cell line (2 weeks), the proportion of intracellular mycoplasmas reached a maximum of 10% and M. hominis-filled protrusions of the host cell membrane were seen by confocal microscopy, suggesting exocytotic dissemination. Of the 1972 regulated host genes, components of the ECM-receptor interaction pathway and phagosome-related integrins were markedly increased. The immune response was quite different to that at the beginning of infection, with a prominent induction of IL1B gene expression, affecting pathways of MAPK signalling, and genes connected with cytokine-cytokine interactions and apoptosis. These data show for the first time the complex, time-dependent reaction of the host directed at mycoplasmal clearance and the counter measures of

Significance of Blastocystis hominis in patients referred for ...

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Objective: To determine the prevalence and related clinical manifestation of B. hominis infection in patients referred for bacteriological ... with a worldwide distribution. However ... abdominal pain and peripheral blood eosinophilia. However ...

Development of real-time PCR for detection of Mycoplasma hominis

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Baczynska, A.; Svenstrup, Helle Friis; Fedder, J.

2004-01-01

BACKGROUND: Mycoplasma hominis is associated with pelvic inflammatory disease, bacterial vaginosis, post partum fever, sepsis and infections of the central nervous system often leading to serious conditions. Association with development of female infertility has also been suggested, but different....... Information on bacterial load in genital swabs can be obtained. The assay allowed detection of M. hominis in a closed system reducing the risk of contamination by amplicon carry-over......., glyceraldehyde-3-phosphate dehydrogenase (gap), as a target. RESULTS: Real-time PCR was optimized to detect 10 copies of M. hominis PG21 genomic DNA. A fluorescence signal was measured for all 20 other M. hominis isolates, and melting curves analysis showed variations in the melting temperature in agreement...

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

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Caroline Spitz dos Santos

2015-12-01

Full Text Available Abstract: Tritrichomonas foetus, a parasite well known for its significance as a venereally transmitted pathogen in cattle, has been identified as a cause of chronic large bowel diarrhea in domestic cats in many countries of the world. In Brazil, several studies on the diagnosis of bovine trichomoniasis have been performed, but until now, no study was made regarding feline trichomoniasis. Thus, this is the first study to report the occurrence of T. foetus and Pentatrichomonas hominis in cats using morphological and molecular analysis. Feces from 77 cats were examined, four of which (5.2% were positive for the presence of parabasalids. Morphological analysis of stained smears revealed piriform trophozoites showing the three anterior flagella, elongated nucleus and axostyle ending abruptly in fillet, characteristic of T. foetus. In scanning and transmission electron microscopy, identification characters similar to those previously reported for T. foetus were observed. The cultures containing trophozoites were submitted for molecular analysis, which resulted positive for T. foetus DNA using specific primers (TFR3 and TFR4, and all samples were positive and subjected to sequencing in which they showed 99.7-100% similarity with another isolate sequencing of T. foetus (JX960422. Although no trophozoite with consistent morphology of P. hominis has been visualized in the samples, differential diagnosis was performed using specific primers for P. hominis (TH3 and TH5 amplicon. In three of the four samples (3.89% sequencing revealed 100% similarity when compared with another sequence of P. hominis deposited in Genbank (KC623939. Therefore, the present study revealed through the diagnostic techniques employed the simultaneous infection by T. foetus and P. hominis in the feces of cats. However, it was necessary to use more than one technique for the diagnosis of the co-infection. These results demonstrate the importance of a correct diagnosis to allow an

Serological differentiation of microsporidia with special reference to Trachipleistophora hominis

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Cheney S.A.

2001-06-01

Full Text Available Myositis is a common clinical syndrome in advanced stages of AIDS. Trachipleistophora hominis (phylum Microspora has been detected in several cases of painful, immobilising myositis in AIDS patients. Enzyme linked immunosorbent assays (ELISAs and Western blotting of protein profiles separated by SDS PAGE were used to determine whether this species could be detected and differentiated by serology. Sixteen microsporidia, including several species known to infect man and species infecting fish, crustaceans and a mosquito, were used as antigen. Each species had a unique profile of SDS PAGE-separated proteins. In Western blots, mouse antiserum, raised to T. hominis and selected for its high ELISA specificity, bound to antigens ranging from less than 25 kDa to greater than 250 kDa with major bands at 39-44 kDa and 98-150 kDa on T. hominis protein profiles. The serum also recognised some high molecular weight antigens in the profiles of Vavraia culicis, Heterosporis anguillarum, and three species of Pleistophora but none in the remaining genera examined. It was concluded that ELISA and Western blotting could be used to detect and differentiate T. hominis in muscle biopsy tissue from patients with myositis. However, sera from T. hominis infected patients in the terminal stages of AIDS would not be useful for detection of infections because of a sharp decline in antibody level.

Molecular detection of Chlamydia Trachomatis and Mycoplasma Hominis in endometriosis lesions

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F. Azizvakili

2016-12-01

Full Text Available Background: Retrograde of menstrual blood into the peritoneal cavity is one of the accepted theories for initiation of endometriosis although indicated that other factors are involved in pathogenesis. Investigation of infectious agents is important in this regard. Objective: To investigate the presence of bacterial infections; Chlamydia trachomatis and Mycoplasma Hominis as risk factors in endometriosis lesions. Methods: This case-control study was conducted in Sarem Hospital in 2014. DNA was extracted from 90 paraffin-embedded blocks included 40 endometriosis tissue samples, 23 samples of endometrial tissue from the same patients and 27 samples of endometrial tissue of the patients without endometriosis, and molecular analysis were performed using polymerase chain reaction. Results were analyzed by Fisher Exact Test and McNemar Test. Findings: Chlamydia trachomatis infection was seen in 11 (27.5% endometriosis tissue, 3 (13% normal tissue from patients and 10 (37% in patient without endometriosis. Mycoplasma hominis was diagnosed in 11 (27.5% endometriosis tissue, 7 (30.4% of normal tissue from patients and one patient without endometriosis (3.7%. These differences show significant relations between infection with Mycoplasma hominis and endometriosis. Conclusion: The findings of this study did not show significant association between Chlamydia trachomatis infections and endometriosis. However; it seems Mycoplasma hominis infection can increase the risk of endometriosis incidence.

Fatal nosocomial meningitis caused by Mycoplasma hominis in an adult patient: case report and review of the literature

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Sophie Reissier

2016-07-01

Full Text Available Meningitis due to Mycoplasma hominis in adults is rarely described, with only three cases having been reported to date. A case of fatal meningitis in a 39-year-old patient after a neurosurgical procedure for a subarachnoid haemorrhage is reported herein. Identification and treatment were significantly delayed because of the rarity of the aetiology and difficulty identifying this organism with the routinely used conventional methods, such as Gram staining and agar growth on standard agar plates. Clinical procedures and the treatment of ‘culture-negative’ central nervous system infections is a real challenge for clinical microbiologists and clinicians, and M. hominis has to be considered as a potential, although very uncommon, pathogen.

The Mycoplasma hominis vaa gene displays a mosaic gene structure

DEFF Research Database (Denmark)

Boesen, Thomas; Emmersen, Jeppe M. G.; Jensen, Lise T.

1998-01-01

Mycoplasma hominis contains a variable adherence-associated (vaa) gene. To classify variants of the vaa genes, we examined 42 M. hominis isolated by PCR, DNA sequencing and immunoblotting. This uncovered the existence of five gene categories. Comparison of the gene types revealed a modular...

The Case Report of Trigger Finger Improved with Hominis Placenta Pharmacopuncture Treatment

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Jeong-Won Kim

2010-12-01

Full Text Available Objectives : The Purpose of this study is to investigate and report the effectiveness of Hominis Placenta using Pharmacopuncture treatment for trigger finger. Methods : 3 Patients are admitted at Dept. of Oriental Rehabilitation, Bu-Chun Jaseng Oriental Medicine Hospital, diagnosed as Trigger finger and treated with Hominis Placenta Pharmacopuncture. Each cases are measured and assessed by Quinnell's classification of triggering and VAS (Visual Analogue Scale scores. Results : 3 Patients of trigger finger have a different kind of cause and fingers lesion they have, but nodules are not significantly found up, so we could classify all of 3 patients to diffuse type. After treatment of Hominis placenta Pharmacopuncture, spontaneous pain and tenderness, grades of triggering are decreased significantly. We would expect that Hominis placenta Pharmacopuncture has a effect on degenerative diseases of diffuse type's tendon sheath. Conclusions : Trigger finger is generally divided into two stages, inflammatory and degenerative stage, and when degenerative stage, Hominis placenta pharmacopuncture appears to be effective.

Life on arginine for Mycoplasma hominis: clues from its minimal genome and comparison with other human urogenital mycoplasmas.

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Sabine Pereyre

2009-10-01

Full Text Available Mycoplasma hominis is an opportunistic human mycoplasma. Two other pathogenic human species, M. genitalium and Ureaplasma parvum, reside within the same natural niche as M. hominis: the urogenital tract. These three species have overlapping, but distinct, pathogenic roles. They have minimal genomes and, thus, reduced metabolic capabilities characterized by distinct energy-generating pathways. Analysis of the M. hominis PG21 genome sequence revealed that it is the second smallest genome among self-replicating free living organisms (665,445 bp, 537 coding sequences (CDSs. Five clusters of genes were predicted to have undergone horizontal gene transfer (HGT between M. hominis and the phylogenetically distant U. parvum species. We reconstructed M. hominis metabolic pathways from the predicted genes, with particular emphasis on energy-generating pathways. The Embden-Meyerhoff-Parnas pathway was incomplete, with a single enzyme absent. We identified the three proteins constituting the arginine dihydrolase pathway. This pathway was found essential to promote growth in vivo. The predicted presence of dimethylarginine dimethylaminohydrolase suggested that arginine catabolism is more complex than initially described. This enzyme may have been acquired by HGT from non-mollicute bacteria. Comparison of the three minimal mollicute genomes showed that 247 CDSs were common to all three genomes, whereas 220 CDSs were specific to M. hominis, 172 CDSs were specific to M. genitalium, and 280 CDSs were specific to U. parvum. Within these species-specific genes, two major sets of genes could be identified: one including genes involved in various energy-generating pathways, depending on the energy source used (glucose, urea, or arginine and another involved in cytadherence and virulence. Therefore, a minimal mycoplasma cell, not including cytadherence and virulence-related genes, could be envisaged containing a core genome (247 genes, plus a set of genes required for

Meningitis in a Chinese adult patient caused by Mycoplasma hominis: a rare infection and literature review

OpenAIRE

Zhou, Menglan; Wang, Peng; Chen, Sharon; Du, Bin; Du, Jinlong; Wang, Fengdan; Xiao, Meng; Kong, Fanrong; Xu, Yingchun

2016-01-01

Background Mycoplasma hominis, a well known cause of neonatal infection, has been reported as a pathogen in urogenital infections in adults; however, central nervous system (CNS) infections are rare. We report here the first case of M. hominis meningitis in China, post neurosurgical treatment for an intracerebral haemorrhage in a 71-year-old male. Case presentation We describe a 71-year-old man who developed M. hominis meningitis after neurosurgical treatment and was successfully treated with...

A comparison between hominy chop and defatted maize germ meal ...

African Journals Online (AJOL)

Defatted maize germ meal (DMG) is arbitrarily rated at a lower economic value than maize meal or hominy chop (HC). Five treatments with 15 steers each were fed different inclusion levels of DMG (0%, 25%, 50%, 75% and 100%), replacing hominy chop during the fattening period. Slaughter data were collected for carcass ...

Epidemiological investigation and antimicrobial susceptibility analysis of ureaplasma species and Mycoplasma hominis in outpatients with genital manifestations.

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Song, Tiejun; Ye, Aiqing; Xie, Xinyou; Huang, Jun; Ruan, Zhi; Kong, Yingying; Song, Jingjuan; Wang, Yue; Chen, Jiangzhong; Zhang, Jun

2014-09-01

The aim of this study was to assess the prevalence and drug resistance of Ureaplasma species and Mycoplasma hominis in outpatients with genital manifestation from 2005 to 2013 in Hangzhou, China. A total of 2689 female and 2336 male patients with various genital symptoms were included in this study. Species identification and antimicrobial susceptibility test were performed by using the mycoplasma IST-2 kit. The prevalence rate of Ureaplasma species was 39.9%, M hominis was 1.2% in female patients, and the coinfection rate was 13.4%; while in males, the prevalence rate of Ureaplasma species was 18.8%, M hominis was 0.4%, and the coinfection rate was 2.9%. Moreover, significantly high positive rates for mycoplasmas (Ureaplasma species M hominis) and were found in 16–20-year-old females (65.2%) and males (27.3%). Ureaplasma species and M hominis displayed relatively lower resistance rates (Ureaplasma species to quinolones (ofloxacin and ciprofloxacin) were much higher (>50%) and increased significantly from 2005 to 2013. Our study indicates that high positive rates of Ureaplasma species and M hominis were found in young outpatients with genital symptoms, and monitoring the local drug resistance is critical for prevention of the occurrence of resistant strains.

Emergence of Cryptosporidium hominis Monkey Genotype II and Novel Subtype Family Ik in the Squirrel Monkey (Saimiri sciureus) in China.

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Liu, Xuehan; Xie, Na; Li, Wei; Zhou, Ziyao; Zhong, Zhijun; Shen, Liuhong; Cao, Suizhong; Yu, Xingming; Hu, Yanchuan; Chen, Weigang; Peng, Gangneng

2015-01-01

A single Cryptosporidium isolate from a squirrel monkey with no clinical symptoms was obtained from a zoo in Ya'an city, China, and was genotyped by PCR amplification and DNA sequencing of the small-subunit ribosomal RNA (SSU rRNA), 70-kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein, and actin genes. This multilocus genetic characterization determined that the isolate was Cryptosporidium hominis, but carried 2, 10, and 6 nucleotide differences in the SSU rRNA, HSP70, and actin loci, respectively, which is comparable to the variations at these loci between C. hominis and the previously reported monkey genotype (2, 3, and 3 nucleotide differences). Phylogenetic studies, based on neighbor-joining and maximum likelihood methods, showed that the isolate identified in the current study had a distinctly discordant taxonomic status, distinct from known C. hominis and also from the monkey genotype, with respect to the three loci. Restriction fragment length polymorphisms of the SSU rRNA gene obtained from this study were similar to those of known C. hominis but clearly differentiated from the monkey genotype. Further subtyping was performed by sequence analysis of the gene encoding the 60-kDa glycoprotein (gp60). Maximum homology of only 88.3% to C. hominis subtype IdA10G4 was observed for the current isolate, and phylogenetic analysis demonstrated that this particular isolate belonged to a novel C. hominis subtype family, IkA7G4. This study is the first to report C. hominis infection in the squirrel monkey and, based on the observed genetic characteristics, confirms a new C. hominis genotype, monkey genotype II. Thus, these results provide novel insights into genotypic variation in C. hominis.

Association of Mycoplasma hominis and Ureaplasma urealyticum with some indicators of nonspecific vaginitis.

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Cedillo-Ramírez, L; Gil, C; Zago, I; Yáñez, A; Giono, S

2000-01-01

The purpose of this study was to determine the isolation rates of Mycoplasma hominis and Ureaplasma urealyticum from three populations of women and also to relate the presence of these microorganisms with some indicators of nonspecific vaginitis. Three hundred vaginal swabs were taken from delivery, pregnant and control (not pregnant) women. Cultures were done in E broth supplemented with arginine or urea. M. hominis was isolated in 5% at delivery, 12% from pregnant and 5% from control women and U. urealyticum was isolated in 21%, 31% and 28% respectively. There was statistical difference in the isolation rate of M. hominis in pregnant women respect to the other groups. Both microorganisms were more frequently isolated in women with acid vaginal pH, amine-like odor in KOH test, clue cells and leucorrhea. M. hominis was isolated in 17% and U. urealyticum in 52% from women with nonspecific vaginitis. M. hominis was isolated in 2% and U. urealyticum in 13% from women without nonspecific vaginitis. Although the presence of clue cells and amine-like odor in KOH test have relationship with Gardnerella vaginalis, these tests could also suggest the presence of these mycoplasmas.

[Pyomyositis, sacroiliitis and spondylodiscitis caused by Staphylococcus hominis in a immunocompetent woman].

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Gómez Rodríguez, N; Durán Muñoz, O

2006-12-01

In absence of risk factors, osteoarticular infections by coagulase-negative staphylococci are very infrequent. We described the case of a immunocompetent 73-year-old-woman that suffered pyomyositis, left sacroiliitis and spondylodiscitis involving the first and second thoracic vertebrae by Staphylococcus hominis. This multifocal infection occurred five-weeks after intramuscular administration of NSAI for treatment of low back pain associated with a herniated disc L4-L5. This is the first know case of a multifocal muscle skeletal infection by Staphylococcus hominis in a patient immunocompetent.

Ureaplasma species and Mycoplasma hominis in cervical fluid of pregnancies complicated by preterm prelabor rupture of membranes.

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Musilova, Ivana; Pliskova, Lenka; Kutova, Radka; Hornychova, Helena; Jacobsson, Bo; Kacerovsky, Marian

2016-01-01

To evaluate Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid and their association with microbial invasion of the amniotic cavity (MIAC) and/or histological chorioamnionitis (HCA) in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). A prospective study of 68 women with singleton pregnancies complicated by PPROM between 24(0/7) and 36(6/7) weeks was conducted. Cervical fluid and amniotic fluid were collected from all women at the time of admission. The Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid were identified using specific real-time PCR. Ureaplasma species and Mycoplasma hominis DNA were identified in 59% (40/69) of the cervical fluid samples. Women with the presence of Ureaplasma species DNA with and without Mycoplasma hominis DNA in the cervical fluid had a higher rate of MIAC alone [35% (14/40) versus 11% (3/28); p = 0.02] and a higher rate of the presence of both MIAC and HCA [30% (12/40) versus 4% (1/28); p = 0.01] than women without Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid. The presence of Ureaplasma species DNA with and without Mycoplasma hominis DNA in the cervical fluid is associated with a higher risk of MIAC or MIAC and HCA together in pregnancies complicated by PPROM.

Homo homini : [luuletused] / Valeria Ränik

Index Scriptorium Estoniae

Ränik, Valeria, 1964-

2004-01-01

Sisu: Homo homini ; "Tegelikult on täiuslik kõik..." ; Mõned kõned ; "Vihmadest lekib lagi..." ; "Sündisid siia, et maksta maksu..." ; "Mingis kohas, mingil ajal..." ; "Emajõgi, Amme jõgi..." ; Kaktus ; Eraelamus ; "Minevikule vesi peale..."

High rates of double-stranded RNA viruses and Mycoplasma hominis in Trichomonas vaginalis clinical isolates in South Brazil.

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da Luz Becker, Débora; dos Santos, Odelta; Frasson, Amanda Piccoli; de Vargas Rigo, Graziela; Macedo, Alexandre José; Tasca, Tiana

2015-08-01

Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil. Copyright © 2015 Elsevier B.V. All rights reserved.

Mycoplasma hominis and Mycoplasma genitalium in the Vaginal Microbiota and Persistent High-Risk Human Papillomavirus Infection

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Sally N. Adebamowo

2017-06-01

Full Text Available BackgroundRecent studies have suggested that the vaginal microenvironment plays a role in persistence of high-risk human papillomavirus (hrHPV infection and thus cervical carcinogenesis. Furthermore, it has been shown that some mycoplasmas are efficient methylators and may facilitate carcinogenesis through methylation of hrHPV and cervical somatic cells. We examined associations between prevalence and persistence of Mycoplasma spp. in the vaginal microbiota, and prevalent as well as persistent hrHPV infections.MethodsWe examined 194 Nigerian women who were tested for hrHPV infection using SPF25/LiPA10 and we identified Mycoplasma genitalium and Mycoplasma hominis in their vaginal microbiota established by sequencing the V3–V4 hypervariable regions of the 16S rRNA gene. We defined the prevalence of M. genitalium, M. hominis, and hrHPV based on positive result of baseline tests, while persistence was defined as positive results from two consecutive tests. We used exact logistic regression models to estimate associations between Mycoplasma spp. and hrHPV infections.ResultsThe mean (SD age of the study participants was 38 (8 years, 71% were HIV positive, 30% M. genitalium positive, 45% M. hominis positive, and 40% hrHPV positive at baseline. At follow-up, 16% of the women remained positive for M. genitalium, 30% for M. hominis, and 31% for hrHPV. There was a significant association between persistent M. hominis and persistent hrHPV (OR 8.78, 95% CI 1.49–51.6, p 0.01. Women who were positive for HIV and had persistent M. hominis had threefold increase in the odds of having persistent hrHPV infection (OR 3.28, 95% CI 1.31–8.74, p 0.008, compared to women who were negative for both.ConclusionWe found significant association between persistent M. hominis in the vaginal microbiota and persistent hrHPV in this study, but we could not rule out reverse causation. Our findings need to be replicated in larger, longitudinal studies and if confirmed

Prevalence and antimicrobial susceptibility of Ureaplasma species and Mycoplasma hominis in Greek female outpatients, 2012-2016.

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Maraki, Sofia; Mavromanolaki, Viktoria Eirini; Nioti, Eleni; Stafylaki, Dimitra; Minadakis, George

2017-11-28

Mycoplasma hominis and Ureaplasma species are opportunistic pathogens associated with urogenital infections, complications during pregnancy and postpartum infections. Appropriate empirical antimicrobial treatment is necessary to achieve an optimal therapeutic outcome. This study evaluated the prevalence and the antimicrobial susceptibility of Mycoplasma hominis and Ureaplasma spp. isolated from 1,008 endocervical samples of outpatients in Crete, Greece, during a five-year period (2012-2016), using the commercially available Mycoview kit (Zeakon diagnostics, France). Ureaplasma spp. was isolated from 116 patients (11.5%), M. hominis from 6 (0.6%), while coinfection with both mycoplasmas was demonstrated in 17 (1.7%). All Ureaplasma strains were susceptible to josamycin and doxycycline. Doxycycline, minocycline and ofloxacin were the most potent antibiotics against M. hominis. Docycycline was proved the most active and is still the drug of choice for the treatment of genital mycoplasma infections. Local surveillance to monitor changes in antimicrobial susceptibilities is necessary to guide treatment strategies.

Isolate resistance of Blastocystis hominis to metronidazole.

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Haresh, K; Suresh, K; Khairul Anus, A; Saminathan, S

1999-04-01

Isolates of Blastocystis hominis from infected immigrant workers from Indonesia, Bangladesh and infected individuals from Singapore and Malaysia were assessed for growth pattern and degree of resistance to different concentrations of metronidazole. Viability of the cells was assessed using eosin-brillian cresyl blue which stained viable cells green and nonviable cells red. The Bangladeshi and Singaporean isolates were nonviable even at the lowest concentration of 0.01 mg/ml, whereas 40% of the initial inoculum of parasites from the Indonesian isolate at day one were still viable in cultures with 1.0 mg/ml metronidazole. The study shows that isolates of B. hominis of different geographical origin have different levels of resistance to metronidazole. The search for more effective drugs to eliminate th parasite appears inevitable, especially since surviving parasites from metronidazole cultures show greater ability to multiply in subcultures than controls.

Clinical efficacy of Saccharomyces boulardii or metronidazole in symptomatic children with Blastocystis hominis infection.

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Dinleyici, Ener Cagri; Eren, Makbule; Dogan, Nihal; Reyhanioglu, Serap; Yargic, Zeynel Abidin; Vandenplas, Yvan

2011-03-01

Although many Blastocystis infections remain asymptomatic, recent data suggest it also causes frequent symptoms. Therapy should be limited to patients with persistent symptoms and a complete workup for alternative etiologies. The goal of this study was to compare the natural evolution (no treatment) to the efficacy of Saccharomyces boulardii (S. boulardii) or metronidazole for the duration of diarrhea and the duration of colonization in children with gastrointestinal symptoms and positive stool examination for Blastocystis hominis. This randomized single-blinded clinical trial included children presenting with gastrointestinal symptoms (abdominal pain, diarrhea, nausea-vomiting, flatulence) more than 2 weeks and confirmed B. hominis by stool examination (B. hominis cysts in the stool with microscopic examination of the fresh stool). The primary end points were clinical evaluation and result of microscopic stool examination at day 15. Secondary end points were the same end points at day 30. Randomization was performed by alternating inclusion: group A, S. boulardii (250 mg twice a day, Reflor®) during 10 days; group B, metronidazole (30 mg/kg twice daily) for 10 days; group C, no treatment. At day 15 and 30 after inclusion, the patients were re-evaluated, and stool samples were examined microscopically. On day 15, children that were still symptomatic and/or were still B. hominis-infected in group C were treated with metronidazole for 10 days. There was no statistically significant difference between the three study groups for age, gender, and the presence of diarrhea and abdominal pain. On day 15, clinical cure was observed in 77.7% in group A (n, 18); in 66.6% in group B (n, 15); and 40% in group C (n:15) (p boulardii has potential beneficial effects in B. hominis infection (symptoms, presence of parasites). These findings challenge the actual guidelines.

The Effects of the Hominis Placenta Herbal acupuncture on Sleep pattern disturbance

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Youn Hyoun-min

2005-02-01

Full Text Available Objective : This study has been designed and performed to identify the effects of Hominis Placenta herbal acupuncture which is usually used in reducing sleep pattern disturbances. Methods : The study subjects studied included 48 patients who were admitted in hospital located in Pusan, and they were classified into 2 groups : 25 patients in the experimental group who injected Hominis Placenta herbal acupuncture and 23 patients in the control group who were treated by acupuncture. The both group injected on GB20, GB12 and HT7 for 5 days without medicine. The sleep pattern disturbance score was measured by using 15 questions according to Korean Sleep Scale A(Oh, Jin Joo. Song, Mi Soon. Kim, Shin Mi. 1998. Results & conclusions : The sleep pattern disturbance score of the experimental group who injected Hominis Placenta herbal acupuncture was significantly lower than that of the control group. (t= 7.00 p= .00 These results provided that Hominis Placenta herbal acupuncture of GB20, GB12 and HT7 was effective for relieving sleep pattern disturbances, it is need more sample's number and more treatmentt's duration.

Mycoplasma hominis: an incidental but significant finding by routine bacteriological culture

DEFF Research Database (Denmark)

Gertsen, Jan Berg; Schønheyder, Henrik Carl

2009-01-01

bacteriological culture during a 4-year period. Methods: Dpt. of Clinical Microbiology, Aalborg Hospital serves a population of 0.5 mio. Aerobic bacteriologic cultures are routinely carried out on 5% horse blood agar and chocolate agar (SSI Diagnostika, DK) at 35 °C in 5% CO2. The finding of translucent, pinpoint...... not respond to surgical drainage of the abscesses (if present) and prolonged empirical intravenous therapy with a β-lactam antibiotic and metronidazole (median 9 days). The tentative diagnosis of M. hominis prompted a change of antibiotic therapy to either moxifloxacin or clindamycin which was followed...... by resolution of symptoms and normalisation of CRP (median 9 days). Conclusion: M. hominis is a rare finding by prolonged incubation of conventional blood agar. A pathogenic role of M. hominis was supported by the lack of clinical response to surgical drainage and prolonged empirical antibiotic therapy...

Molecular design of Mycoplasma hominis Vaa adhesin

DEFF Research Database (Denmark)

Boesen, Thomas; Fedosova, Natalya U.; Kjeldgaard, Morten

2001-01-01

The variable adherence-associated (Vaa) adhesin of the opportunistic human pathogen Mycoplasma hominis is a surface-exposed, membrane-associated protein involved in the attachment of the bacterium to host cells. The molecular masses of recombinant 1 and 2 cassette forms of the protein determined...

Furuncular Myiasis Caused by Dermatobia hominis in a Traveler Returning from the Amazon Jungle.

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Kuşcu, Ferit; Özsoy, Kerem Mazhar; Ulu, Aslıhan; Kurtaran, Behice; Kömür, Süheyla; İnal, Ayşe Seza; Taşova, Yeşim; Aksu, Hasan Salih Zeki

2017-09-01

A 39-year-old man who was returning from the Amazon Jungle and had no medical history presented with a furuncular lesion on his right parietal scalp. Despite receiving appropriate antimicrobial treatment, his lesion did not heal. After surgical intervention, a Dermatobia hominis larva was extracted. The human botfly D. hominis is the most common causative agent of furuncular myiasis among travelers returning from Central and South America. Surgery is the main treatment option, and secondary bacterial infection should be kept in mind.

Mycoplasma hominis: an incidental but significant finding by routine bacteriological culture

DEFF Research Database (Denmark)

Gertsen, Jan Berg; Schønheyder, Henrik Carl

2009-01-01

Objectives: M. hominis is part of the normal mucosal flora and is primarily associated with infections in the genitourinary tract. Most infections occur following delivery or genitourinary instrumentation, but are also seen in immunocompromised patients. We present 4 cases diagnosed by routine ba...... Serum Institut, Copenhagen (by courtesy to Jørgen Skov). Results: The four patients were immunocompetent women (23-56 years of age) without significant comorbidity (Table). In all patients M. hominis were obtained in pure culture. At the time of diagnosis three patients had abscesses...

Dermabacter hominis: a usually daptomycin-resistant gram-positive organism infrequently isolated from human clinical samples

Science.gov (United States)

Fernández-Natal, I; Sáez-Nieto, J A; Medina-Pascual, M J; Albersmeier, A; Valdezate, S; Guerra-Laso, J M; Rodríguez, H; Marrodán, T; Parras, T; Tauch, A; Soriano, F

2013-01-01

During a 12-year period, Dermabacter hominis was isolated from 21 clinical samples belonging to 14 patients attending a tertiary hospital in León, Spain. Samples included blood cultures (14), peritoneal dialysis catheter exit sites (three), cutaneous abscesses (two), an infected vascular catheter (one) and a wound swab (one). Identification was made by API Coryne™ V2.0, Biolog™ GP2 and 16S rRNA gene amplification. Six febrile patients had positive blood cultures (one, two or three sets) and all of them were treated with teicoplanin (two patients), vancomycin, ampicillin plus gentamicin, amoxicillin/clavulanic acid and ciprofloxacin (one each). An additional patient with a single positive blood culture was not treated, the finding being considered non-significant. In the remaining seven patients the organism was isolated from a single specimen and three of them received antimicrobial treatment (ciprofloxacin, ceftriaxone plus vancomycin and amoxicillin/clavulanic acid). At least ten patients had several underlying diseases and conditions, and no direct mortality was observed in relation to the isolated organism. All isolates were susceptible to vancomycin, rifampin and linezolid. Resistance to other antibiotics varied: erythromycin (100%), clindamycin (78.5%), ciprofloxacin (21.4%) and gentamicin, quinupristin-dalfopristin, benzylpenicillin and imipenem 7.1% each. Thirteen isolates were highly resistant to daptomycin with MICs ranging from 8 to 48 (MIC90 = 32 mg/L); only one was daptomycin-sensitive (MIC = 0.19 mg/L). PMID:25356327

Cloning, sequencing and variability analysis of the gap gene from Mycoplasma hominis

DEFF Research Database (Denmark)

Mygind, Tina; Jacobsen, Iben Søgaard; Melkova, Renata

2000-01-01

The gap gene encodes the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The gene was cloned and sequenced from the Mycoplasma hominis type strain PG21(T). The intraspecies variability was investigated by inspection of restriction fragment length polymorphism (RFLP) patterns...... after polymerase chain reaction (PCR) amplification of the gap gene from 15 strains and furthermore by sequencing of part of the gene in eight strains. The M. hominis gap gene was found to vary more than the Escherichia coli counterpart, but the variation at nucleotide level gave rise to only a few...... amino acid substitutions. To verify that the gene was expressed in M. hominis, a polyclonal antibody was produced and tested against whole cell protein from 15 strains. The enzyme was expressed in all strains investigated as a 36-kDa protein. All strains except type strain PG21(T) showed reaction...

The use of enzyme-linked immunosorbent assay for detection of Mycoplasma hominis antibodies in infertile women serum samples

DEFF Research Database (Denmark)

Baczynska, Agata; Friis Svenstrup, Helle; Fedder, Jens

2005-01-01

BACKGROUND: Besides Chlamydiae trachomatis and Mycoplasma genitalium, Mycoplasma hominis may also cause infertility due to damage of the Fallopian tubes. Therefore serum samples from infertile women were analyzed for antibodies to M. hominis. METHODS: Sera from 304 infertile women were investigat...

An infant case with hydrocephalus as the initial manifestation of Mycoplasma hominis-associated meningitis.

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Taku, Keisuke; Hoshina, Takayuki; Haro, Kaoru; Ichikawa, Shun; Kinjo, Tadamune; Takahashi, Mayu; Akiba, Daisuke; Fukuda, Kazumasa; Kusuhara, Koichi

2017-10-01

We report an infant with hydrocephalus as the initial manifestation of Mycoplasma hominis-associated meningitis, who recovered without appropriate antimicrobial treatment. The analysis of the 16S rRNA gene by polymerase chain reaction amplification using universal primers and pathogen-specific primers was useful for the diagnosis and the investigation of serial detection status of the pathogen. This method may be helpful for the assessment of the frequency and the prediction of severity in M. hominis-associated central nervous system infection in infants, and investigating the association between M. hominis and the development of hydrocephalus. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Genetic Passive Immunization with Adenoviral Vector Expressing Chimeric Nanobody-Fc Molecules as Therapy for Genital Infection Caused by Mycoplasma hominis.

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Daria A Burmistrova

Full Text Available Developing pathogen-specific recombinant antibody fragments (especially nanobodies is a very promising strategy for the treatment of infectious disease. Nanobodies have great potential for gene therapy application due to their single-gene nature. Historically, Mycoplasma hominis has not been considered pathogenic bacteria due to the lack of acute infection and partially due to multiple studies demonstrating high frequency of isolation of M. hominis samples from asymptomatic patients. However, recent studies on the role of latent M. hominis infection in oncologic transformation, especially prostate cancer, and reports that M. hominis infects Trichomonas and confers antibiotic resistance to Trichomonas, have generated new interest in this field. In the present study we have generated specific nanobody against M. hominis (aMh, for which the identified target is the ABC-transporter substrate-binding protein. aMh exhibits specific antibacterial action against M. hominis. In an attempt to improve the therapeutic properties, we have developed the adenoviral vector-based gene therapy approach for passive immunization with nanobodies against M. hominis. For better penetration into the mucous layer of the genital tract, we fused aMh with the Fc-fragment of IgG. Application of this comprehensive approach with a single systemic administration of recombinant adenovirus expressing aMh-Fc demonstrated both prophylactic and therapeutic effects in a mouse model of genital M. hominis infection.

Relevant prevalence of Mycoplasma hominis and Ureaplasma urealyticum serogroups in HIV-1 infected men without urethritis symptoms

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CORDOVA Caio Mauricio Mendes

2000-01-01

Full Text Available M. hominis and U. urealyticum are the better-known mycoplasma species pathogenic to the human genitourinary tract, causing mainly urethritis, bacterial vaginosis and pregnancy complications. In HIV-infected patients, the prevalence and role of these species is still not well known. The aim of this work was to determinate the prevalence of these species in this group of male patients (HIV group, in comparison to a group of men with clinical symptoms of urethritis (STD group. M. hominis was isolated from 7.5% patients (8/106 and U. urealyticum from 18.9% patients (20/106 from the HIV group, being among these 62.5% and 85% in significant concentrations, respectively. In the STD group these rates were 0.9% (1/110 for M. hominis and 13.6% (15/110 for U. urealyticum, being 100% and 93.3% in significant concentrations, respectively. We could demonstrate infection rates by these mycoplasma species in the HIV group as high as the one found in the STD one, what may indicate the occurrence of opportunistic infections in our population. This fact is discussed here because in immunosuppressed patients, specially M. hominis has been reported causing severe infections, even systemically.

Genetic recombination and Cryptosporidium hominis virulent subtype IbA10G2.

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Li, Na; Xiao, Lihua; Cama, Vitaliano A; Ortega, Ynes; Gilman, Robert H; Guo, Meijin; Feng, Yaoyu

2013-10-01

Little is known about the emergence and spread of virulent subtypes of Cryptosporidium hominis, the predominant species responsible for human cryptosporidiosis. We conducted sequence analyses of 32 genetic loci of 53 C. hominis specimens isolated from a longitudinally followed cohort of children living in a small community. We identified by linkage disequilibrium and recombination analyses only limited genetic recombination, which occurred exclusively within the 60-kDa glycoprotein gene subtype IbA10G2, a predominant subtype for outbreaks in industrialized nations and a virulent subtype in the study community. Intensive transmission of virulent subtype IbA10G2 in the study area might have resulted in genetic recombination with other subtypes. Moreover, we identified selection for IbA10G2 at a 129-kb region around the 60-kDa glycoprotein gene in chromosome 6. These findings improve our understanding of the origin and evolution of C. hominis subtypes and the spread of virulent subtypes.

Microbial load of umbilical cord blood Ureaplasma species and Mycoplasma hominis in preterm prelabor rupture of membranes.

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Kacerovsky, Marian; Pliskova, Lenka; Menon, Ramkumar; Kutova, Radka; Musilova, Ivana; Maly, Jan; Andrys, Ctirad

2014-11-01

To evaluate Ureaplasma species and M. hominis DNA in the umbilical cord blood and its correlation with its microbial load in the amniotic fluid, as a measure of microbial burden in fetal inflammatory response and neonatal outcome in pregnancies complicated by preterm prelabor rupture of membranes (pPROM). A retrospective study of 158 women with singleton pregnancies complicated by pPROM between 24(0/7) and 36(6/7) weeks was conducted. Amniotic fluid was obtained from all women by transabdominal amniocentesis, and umbilical cord blood was obtained by venipuncture from umbilical cords immediately after the delivery of the neonates. The Ureaplasma species and M. hominis DNA was quantitated using absolute quantification techniques. Ureaplasma species and M. hominis DNA was identified in 9% of the umbilical cord blood samples. No correlation between the amniotic fluid and umbilical cord blood microbial load was observed. The presence of Ureaplasma species and M. hominis DNA in the umbilical cord blood had no impact on short-term neonatal morbidity. A high microbial load of genital mycoplasma Ureaplasma species DNA in the umbilical cord in pregnancies complicated by pPROM is not associated with a high fetal inflammatory response and is therefore not associated with serious neonatal morbidity.

The infection of Mycoplasma hominis after total knee replacement: Case report and literature review

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Hong-Jiu Qiu

2017-08-01

Full Text Available The Mycoplasma hominis infection is a rare postoperative complication after joint replacement. Based on our knowledge, there were only two cases reported by Korea all over the world currently. A case of postoperative Mycoplasma hominis infection after total knee replacement in our hospital was reported in this article. It was confirmed through mass spectrometer and Mycoplasma cultivation and treated by the first stage debridement, polyethylene insert replacement, and then drainage and irrigation combined with sensitive antibiotics after the operation. We observed that the C reactive protein (CRP level correlates with the development of disease, while the erythrocyte sedimentation rate (ESR remains at a high level, indicating the relevance between the Mycoplasma hominis infection caused by knee joint replacement and CRP. This study aims to report the case and review relevant literature.

Detection of Tritrichomonas foetus and Pentatrichomonas hominis in intestinal tissue specimens of cats by chromogenic in situ hybridization.

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Mostegl, Meike M; Wetscher, Andreas; Richter, Barbara; Nedorost, Nora; Dinhopl, Nora; Weissenböck, Herbert

2012-02-10

In this retrospective study 102 cats were analyzed for the presence of trichomonads in intestinal tissue sections using chromogenic in situ hybridization (CISH). Two intestinal trichomonad species are described in cats: Pentatrichomonas hominis and Tritrichomonas foetus. While P. hominis is considered a mere commensal, T. foetus has been found to be the causative agent of feline large-bowel diarrhea. For the detection of both agents within intestinal tissue CISH assays using three different probes were performed. In the first CISH run a probe specific for all relevant members of the order Trichomonadida (OT probe) was used. In a second CISH run all positive samples were further examined on three consecutive tissue sections using the OT probe, a probe specific for the family of Tritrichomonadidae (Tritri probe) and a newly designed probe specifically detecting P. hominis (Penta hom probe). In total, four of the 102 cats were found to be positive with the OT probe. Thereof, one cat gave a positive reaction with the P. hominis probe and three cats were positive with the T. foetus probe. All Trichomonas-positive cats were pure-bred and between 8 and 32 weeks of age. In one cat positive for T. foetus large amounts of parasites were found in the gut lumen and invading the intestinal mucosa. The species of the detected trichomonads were confirmed by polymerase chain reaction and nucleotide sequencing of a part of the 18S ribosomal RNA gene. In this study, the usefulness of CISH to detect intestinal trichomonads within feline tissue samples was shown. Additionally, the specific detection of P. hominis using CISH was established. Generally, it was shown that CISH is well suited for detection and differentiation of trichomonosis in retrospective studies using tissue samples. Copyright © 2011 Elsevier B.V. All rights reserved.

Cloning, sequencing and variability analysis of the gap gene from Mycoplasma hominis

DEFF Research Database (Denmark)

Mygind, Tina; Jacobsen, Iben Søgaard; Melkova, Renata

2000-01-01

The gap gene encodes the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The gene was cloned and sequenced from the Mycoplasma hominis type strain PG21(T). The intraspecies variability was investigated by inspection of restriction fragment length polymorphism (RFLP) patterns...... after polymerase chain reaction (PCR) amplification of the gap gene from 15 strains and furthermore by sequencing of part of the gene in eight strains. The M. hominis gap gene was found to vary more than the Escherichia coli counterpart, but the variation at nucleotide level gave rise to only a few...

[Standardization of Blastocystis hominis diagnosis using different staining techniques].

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Eymael, Dayane; Schuh, Graziela Maria; Tavares, Rejane Giacomelli

2010-01-01

The present study was carried out from March to May 2008, with the aim of evaluating the effectiveness of different techniques for diagnosing Blastocystis hominis in a sample of the population attended at the Biomedicine Laboratory of Feevale University, Novo Hamburgo, Rio Grande do Sul. On hundred feces samples from children and adults were evaluated. After collection, the samples were subjected to the techniques of spontaneous sedimentation (HPJ), sedimentation in formalin-ether (Ritchie) and staining by means of Gram and May-Grünwald-Giemsa (MGG). The presence of Blastocystis hominis was observed in 40 samples, when staining techniques were used (MGG and Gram), while sedimentation techniques were less efficient (32 positive samples using the Ritchie technique and 20 positive samples using the HPJ technique). Our results demonstrate that HPJ was less efficient than the other methods, thus indicating the need to include laboratory techniques that enable parasite identification on a routine basis.

Electrophoretic analysis of proteins from Mycoplasma hominis strains detected by SDS-PAGE, two-dimensional gel electrophoresis and immunoblotting

DEFF Research Database (Denmark)

Andersen, H; Birkelund, Svend; Christiansen, Gunna

1987-01-01

The proteins of 14 strains of Mycoplasma hominis were compared by SDS-PAGE in gradient gels, by two-dimensional (2D) gel electrophoresis of extracts of 35S-labelled cells and by immunoblot analysis of cell proteins. The strains examined included the M. hominis type strain PG21 and 13 others...... isolated variously from genital tract, mouth, blood, upper urinary tract and a wound. These 14 strains shared 76-99% of proteins in SDS-gradient gel analysis and 41-72% in the 2D gels. As expected, the immunoblot analysis likewise revealed the existence of an extensive common protein pattern in M. hominis...

Gastrodiscoides hominis infection in a Nigerian- case report | Dada ...

African Journals Online (AJOL)

Il s'agit d'un rapport d'un cas d'un enfant Nigerian âgé de sept ans qui s'est présenté atteints des traits de la sous alimentation et de l'anénue également atteint des gastrodiscoides hominis et ascaris lumbricoides. A la suite de la clairance des vers on a remarqué une enorme amélioration dans le statut de la santé de cet ...

[Usefulness of conventional polymerase chain reaction for the detection of Mycoplasma hominis, Ureaplasma spp. and Trichomonas vaginalis in female outpatient's genital samples].

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Alarcón, Gonzalo; Barraza, Gabriela; Vera, Andrea; Wozniak, Aniela; García, Patricia

2016-02-01

Trichomonas vaginalis, Mycoplasma hominis and Ureaplasma spp. are microorganisms responsible for genitourinary and pregnancy pathologies. Nucleic acid amplification methods have shown several advantages, but have not been widely studied for the detection of these microorganisms. To implement a conventional polymerase chain reaction (PCR) for the detection of the microorganisms and to compare its results versus the methods currently used at our laboratory. 91 available samples were processed by PCR, culture (M. hominis y Ureaplasma spp.) and wet mount (T vaginalis). Results were compared and statistically analyzed by kappa agreement test. 85, 80 and 87 samples resulted in agreement for the detection of M. hominis, Ureaplasma spp. y T. vaginalis, respectively. For M. hominis and Ureaplasma spp., agreement was substantial, whereas for T. vaginalis it was moderate, however, for the latter, PCR detected more cases than wet mount. We recommend the implementation of PCR for detection of T. vaginalis whereas culture kit is still a useful method for the other microorganisms.

Antigenic and genomic homogeneity of successive Mycoplasma hominis isolates

DEFF Research Database (Denmark)

Jensen, Lise Torp; Thorsen, P; Møller, B

1998-01-01

Sixty Mycoplasma hominis isolates were obtained from the cervices of pregnant women and from the ears or pharynges of their newborn babies. The isolates were examined by SDS-PAGE and pulsed-field gel electrophoresis. Antigenic and genomic profiles were obtained for 16 series with two or more...

Antimicrobial susceptibility patterns of Ureaplasma species and Mycoplasma hominis in pregnant women.

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Redelinghuys, Mathys J; Ehlers, Marthie M; Dreyer, Andries W; Lombaard, Hennie A; Kock, Marleen M

2014-03-28

Genital mycoplasmas colonise up to 80% of sexually mature women and may invade the amniotic cavity during pregnancy and cause complications. Tetracyclines and fluoroquinolones are contraindicated in pregnancy and erythromycin is often used to treat patients. However, increasing resistance to common antimicrobial agents is widely reported. The purpose of this study was to investigate antimicrobial susceptibility patterns of genital mycoplasmas in pregnant women. Self-collected vaginal swabs were obtained from 96 pregnant women attending an antenatal clinic in Gauteng, South Africa. Specimens were screened with the Mycofast Revolution assay for the presence of Ureaplasma species and Mycoplasma hominis. The antimicrobial susceptibility to levofloxacin, moxifloxacin, erythromycin, clindamycin and tetracycline were determined at various breakpoints. A multiplex polymerase chain reaction assay was used to speciate Ureaplasma positive specimens as either U. parvum or U. urealyticum. Seventy-six percent (73/96) of specimens contained Ureaplasma spp., while 39.7% (29/73) of Ureaplasma positive specimens were also positive for M. hominis. Susceptibilities of Ureaplasma spp. to levofloxacin and moxifloxacin were 59% (26/44) and 98% (43/44) respectively. Mixed isolates (Ureaplasma species and M. hominis) were highly resistant to erythromycin and tetracycline (both 97% resistance). Resistance of Ureaplasma spp. to erythromycin was 80% (35/44) and tetracycline resistance was detected in 73% (32/44) of Ureaplasma spp. Speciation indicated that U. parvum was the predominant Ureaplasma spp. conferring antimicrobial resistance. Treatment options for genital mycoplasma infections are becoming limited. More elaborative studies are needed to elucidate the diverse antimicrobial susceptibility patterns found in this study when compared to similar studies. To prevent complications in pregnant women, the foetus and the neonate, routine screening for the presence of genital mycoplasmas is

Interaction of Mycoplasma hominis PG21 with Human Dendritic Cells: Interleukin-23-Inducing Mycoplasmal Lipoproteins and Inflammasome Activation of the Cell.

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Goret, J; Béven, L; Faustin, B; Contin-Bordes, C; Le Roy, C; Claverol, S; Renaudin, H; Bébéar, C; Pereyre, S

2017-08-01

Mycoplasma hominis lacks a cell wall, and lipoproteins anchored to the extracellular side of the plasma membrane are in direct contact with the host components. A Triton X-114 extract of M. hominis enriched with lipoproteins was shown to stimulate the production of interleukin-23 (IL-23) by human dendritic cells (hDCs). The inflammasome activation of the host cell has never been reported upon M. hominis infection. We studied here the interaction between M. hominis PG21 and hDCs by analyzing both the inflammation-inducing mycoplasmal lipoproteins and the inflammasome activation of the host cell. IL-23-inducing lipoproteins were determined using a sequential extraction strategy with two nondenaturing detergents, Sarkosyl and Triton X-114, followed by SDS-PAGE separation and mass spectrometry identification. The activation of the hDC inflammasome was assessed using PCR array and enzyme-linked immunosorbent assay (ELISA). We defined a list of 24 lipoproteins that could induce the secretion of IL-23 by hDCs, 5 with a molecular mass between 20 and 35 kDa and 19 with a molecular mass between 40 and 100 kDa. Among them, lipoprotein MHO_4720 was identified as potentially bioactive, and a synthetic lipopeptide corresponding to the N-terminal part of the lipoprotein was subsequently shown to induce IL-23 release by hDCs. Regarding the hDC innate immune response, inflammasome activation with caspase-dependent production of IL-1β was observed. After 24 h of coincubation of hDCs with M. homini s, downregulation of the NLRP3-encoding gene and of the adaptor PYCARD-encoding gene was noticed. Overall, this study provides insight into both protagonists of the interaction of M. hominis and hDCs. IMPORTANCE Mycoplasma hominis is a human urogenital pathogen involved in gynecologic and opportunistic infections. M. hominis lacks a cell wall, and its membrane contains many lipoproteins that are anchored to the extracellular side of the plasma membrane. In the present study, we focused on

Two crystal structures of dihydrofolate reductase-thymidylate synthase from Cryptosporidium hominis reveal protein–ligand interactions including a structural basis for observed antifolate resistance

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Anderson, Amy C., E-mail: aca@dartmouth.edu [Dartmouth College, Department of Chemistry, Burke Laboratories, Hanover, NH 03755 (United States)

2005-03-01

An analysis of the protein–ligand interactions in two crystal structures of DHFR-TS from C. hominis reveals a possible structural basis for observed antifolate resistance in C. hominis DHFR. A comparison with the structure of human DHFR reveals residue substitutions that may be exploited for the design of species-selective inhibitors. Cryptosporidium hominis is a protozoan parasite that causes acute gastrointestinal illness. There are no effective therapies for cryptosporidiosis, highlighting the need for new drug-lead discovery. An analysis of the protein–ligand interactions in two crystal structures of dihydrofolate reductase-thymidylate synthase (DHFR-TS) from C. hominis, determined at 2.8 and 2.87 Å resolution, reveals that the interactions of residues Ile29, Thr58 and Cys113 in the active site of C. hominis DHFR provide a possible structural basis for the observed antifolate resistance. A comparison with the structure of human DHFR reveals active-site differences that may be exploited for the design of species-selective inhibitors.

Analysis of a Mycoplasma hominis membrane protein, P120

DEFF Research Database (Denmark)

Christiansen, Gunna; Mathiesen, SL; Nyvold, Charlotte Guldborg

1994-01-01

The monoclonal antibody mAb 26.7D generated against a clinical isolate of Mycoplasma hominis 7488 was shown to react with a surface-exposed epitope on a 120-kDa protein (P120). The gene encoding the protein was cloned and sequenced, and the transcriptional start point was determined by primer...

Freqüência de infecção pelo Mycoplasma hominis e Ureaplasma urealyticum em mulheres inférteis e relação com repercussões clínicas Frequency of infection with Mycoplasma hominis and Ureaplasma urealyticum in infertile women and clinical repercussions

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Ivan Araujo Penna

2005-02-01

.9% in the infertile women and 13.8% in the control group. Among the infertile patients, a relationship between the presence of the two agents and changes in the histerosalpingography result (OR: 3.20; IC 95%: 1.05-9.73, presence of dyspareunia (OR: 10.72; IC 95%: 3.21-35.77 and vaginal discharge (OR: 8.5; IC 95%: 2.83-26.02, besides endocervical culture positive for Escherichia coli (OR: 6.09; IC 95%: 4.95-52.25 was observed. CONCLUSION: Mycoplasma hominis and Ureaplasma urealyticum infection rate is low in infertile patients and is associated with reproductive sequels.

Complete Genome Sequence of Mycoplasma hominis Strain Sprott (ATCC 33131), Isolated from a Patient with Nongonococcal Urethritis.

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Calcutt, Michael J; Foecking, Mark F

2015-07-09

Presented here is the complete and annotated genome sequence of Mycoplasma hominis Sprott (ATCC 33131). The chromosome comprises 695,214 bp, which is approximately 30 kb larger than the syntenic genome of M. hominis PG21(T). Tetracycline resistance of strain Sprott is most probably conferred by the tetM determinant, harbored on a mosaic transposon-like structure. Copyright © 2015 Calcutt and Foecking.

Microbiological and molecular characterization of human clinical isolates of Staphylococcus cohnii, Staphylococcus hominis, and Staphylococcus sciuri.

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Garza-González, Elvira; Morfin-Otero, Rayo; Martínez-Vázquez, Manuel A; Gonzalez-Diaz, Esteban; González-Santiago, Omar; Rodríguez-Noriega, Eduardo

2011-12-01

The incidence of coagulase-negative staphylococci reported as causative agents of nosocomial infections has risen in the last decade. The aim of this study was to characterize biofilm formation, antibiotic resistance, SCCmec type, and genetic relatedness in clinical isolates of Staphylococcus cohnii, Staphylococcus hominis, and Staphylococcus sciuri recovered from humans. Clinically relevant isolates of S. cohnii (n = 15), S. hominis (n = 9), and S. sciuri (n = 6), were collected from patients. Biofilm formation was evaluated using crystal violet staining, drug susceptibility was assessed using the broth microdilution method, and methicillin resistance was measured using the cefoxitin disk test. SCCmec was typed using 2 different methodologies, and genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE). Sixty percent (9/15) of S. cohnii, 33% (3/9) of S. hominis, and 50% (3/6) of S. sciuri isolates were categorized as weak producers of biofilm. None of the isolates were resistant to vancomycin or linezolid. All 3 species showed a high resistance (> 66%) to ampicillin, levofloxacin, erythromycin, and ceftriaxone, and the majority of the isolates were methicillin-resistant. PFGE revealed that the S. cohnii isolates comprised 1 dominant clone. The S. cohnii, S. hominis, and S. sciuri isolates analyzed in this study showed a high methicillin resistance and resistance to other antimicrobials. The results of this study strongly suggest that coagulase-negative staphylococci harbour new SCCmec elements. We report the first case of a clone of S. cohnii associated with human disease.

Resposta imune-humoral e celular em bovinos da raça Nelore imunizados com extrato de larvas (L2 e L3 de Dermatobia hominis (Linnaeus Jr., 1781 Immune humoral and cellular response of nelore bovines immunized with larvae extract (L2 and L3 of Dermatobia hominis (Linnaeus Jr., 1781

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Nelson Luis Mello Fernandes

2007-06-01

, making it difficult to industrial use. Nowadays, the chemical control is utilized against dermatobiosis, therefore it leads to rising toxic chemicals in the animals and environment. The immunological challenge with D. hominis larval extract may represent an important altervative for this parasitosis control. Humoral and cellular immune responses were tested in bovine using an antigenic extract prepared with D. hominis larvae. Three groups of 10 months old Nelore females were used. The first group (A received immunogenic larval extract of D. hominis with fifteen-days interval between injections; the group (B was the control and has not received any sort of treatment; and the group (C received an ectoparasitecide treatment based on Dichlorvos associated to Cypermetrina. Aditionally, leucogram and levels of IgG against D. hominis by immunoassay technique were evaluated. As for the humoral immunity, animals from group A presented higher IgG production against D. hominis with maximum levels of circulating antibodies at the 45th day after the first injection. These animals also showed higher production of neutrophils, eosinophils and monocytes than those from groups B and C. The number of D. hominis larvae nodules observed in animals from the group C was 148.3% larger than those from group A and B. The evidence concerning both cellular and humoral immune responses as well as the reduction on nodules number are an indication that the immunization against D. hominis was partially protective for the immunized bovines.

Cryptosporidium y blastocistis hominis como agentes patógenos en el síndrome diarréico

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Ligia I. Moncada

1989-12-01

Full Text Available En una comunidad de escasos recursos de Bogotá se tomaron muestras de heces de niños menores de diez años con diarrea y de niños sin diarrea. Las muestras del grupo de estudio y del grupo control resultaron negativas para el Cryptosporidium. Se encontraron positivas para Blastocistis hominis 17 muestras del grupo de estudio (8.3%. y 5 del grupo control (10.4%. Los síntomas predominantes fueron fiebre, dolor abdominal y pérdida del apetito. El B. hominis se asoció con la Escherichia coli, Salmonella campylobacter, E. histiolytica, Giardia lamblia, Ascaris lumbricoides y rotavírus. No se confirmó el papel que en los últimos años se le atribuye al Cryptosporidium y al B. hominis como agentes productores de diarrea.

Complete Genome Sequence of the Human Gut Symbiont Roseburia hominis

DEFF Research Database (Denmark)

Travis, Anthony J.; Kelly, Denise; Flint, Harry J

2015-01-01

We report here the complete genome sequence of the human gut symbiont Roseburia hominis A2-183(T) (= DSM 16839(T) = NCIMB 14029(T)), isolated from human feces. The genome is represented by a 3,592,125-bp chromosome with 3,405 coding sequences. A number of potential functions contributing to host...

Characteristics of Factors of Protozoa Blastocystis hominis Persistence.

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Potaturkina-Nesterova, N I; Il'ina, N A; Bugero, N V; Nesterov, A S

2016-10-01

Persistence activity manifested in the expression of anti-lysozyme, anti-lactoferrin, and antihistone factors promoting inactivation of natural anti-infection resistance factors in the body was revealed in Blastocystis hominis protozoa. Activities of these factors were ranged. The frequency of these factors in clinical isolates of blastocyst decreased in the following order: anti-lactoferrin activity (84.5±3.7%)→anti-lysozyme activity (64.8±5.7%)→anti-histone activity (48.1±2.3%). In healthy humans, the corresponding parameters were 7.3±1.3, 5.3±0.9, and 3.3±0.4%, respectively (pprotozoa.

Cryptosporidiosis outbreak in a child day-care center caused by an unusual Cryptosporidium hominis subtype.

Science.gov (United States)

Goñi, Pilar; Almagro-Nievas, Diego; Cieloszyk, Joanna; Lóbez, Silvia; Navarro-Marí, José María; Gutiérrez-Fernández, José

2015-12-01

This work describes the genetic characterization of Cryptosporidium and Giardia involved in an outbreak in a nursery school in Granada, Spain, that affected seven children under the age of 4. Nucleic acids were extracted from the seven stool samples positive to Cryptosporidium or Giardia by microscopy and/or immunochromatography. The species and subtypes of Cryptosporidium were identified by PCR-RFLP and PCR of the SSUrRNA and gp60 genes, respectively. The assemblages of Giardia duodenalis isolates were characterized by PCR of the tpi gene. PCR products were sequenced and analyzed. All of the isolates were positive for Cryptosporidium hominis. Five of them belonged to subtype IaA11R2, one to subtype IbA10G2R2, and the other could not be identified. Three of these samples were positive for G. duodenalis by PCR, two belonging to the assemblage A, and the other one to assemblage B. This is the first report of Cryptosporidium hominis subtype IaA11R2 as a cause of an outbreak in Europe where subtype IbA10G2R2 is the most frequently identified. In the case of Giardia, an outbreak could not be confirmed because of the low number of positive samples and the low genetic variability of the amplified fragments for assemblage A of tpi gene. A new subtype, of Cryptosporidium hominis named IaA11R2, has been described as a cause of an outbreak in a nursery school in Granada, Spain. However an outbreak of giardiasis could not be confirmed. Copyright © 2015 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Co-infections with Ureaplasma parvum, Mycoplasma hominis and Chlamydia trachomatis in a human immunodeficiency virus positive woman with vaginal discharge.

Science.gov (United States)

Ghosh, Arnab; Rawre, Jyoti; Khanna, Neena; Dhawan, Benu

2013-01-01

A 30-year-old human immunodeficiency virus (HIV)-1 infected woman presented with vaginal discharge and associated vulval irritation. The vaginal swabs tested positive for Ureaplasma parvum and Mycoplasma hominis by both culture and polymerase chain reaction (PCR). The specimen also tested positive for Chlamydia trachomatis deoxyribonucleic acid (DNA) by cryptic plasmid and omp1 gene PCR assays. The patient was successfully treated with azithromycin based on the antibiotic susceptibility testing results of U. parvum and M. hominis by microbroth dilution. Since sexually transmitted infections enhance the transmission of HIV, HIV-positive patients should be screened routinely for these pathogens.

PCR Research of Mycoplasma hominis in Trichomonas vaginalis Cells of Sichuan Province%四川地区阴道毛滴虫内人型支原体的PCR检测

Institute of Scientific and Technical Information of China (English)

朱晓燕; 王雅静; 毕世樑; 张仁刚

2009-01-01

20 isolates of Trichomonas vaginalis were collected from clinical patients in hospital. After pure cultivation, genomic DNA of T. vaginalis was extracted. A pair of specific 16S rDNA primers was designed based on the sequence of Mycoplasma hominis. M. hominis in T. vaginalis cells were detected by PCR. Altogether 20 isolates of T. vaginalis were collected, 13 out of 20 T. vaginalis isolates harboured M. hominis, and the symbiosis ratio between T. vaginalis and M. hominis was up to 65%. It suggested that the symbiotic relationship between T. vaginalis and M. hominis occurred commonly in Sichuan, China.%从临床上分离获得20株阴道毛滴虫虫株,经纯化培养后,提取基因组DNA.以人型支原体16S rDNA序列设计特异性引物,利用PCR技术检测阴道毛滴虫内的人型支原体,结果有13株为人型支原体阳性,感染率为65%,表明阴道毛滴虫与人型支原体的共生关系在中国四川具有普遍性.

"The rate of Chlamydia Trachomatis, Mycoplasma Hominis and Ureaplasma Urealyticum in females with habitual abortion and its comparison with control group "

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Salari MH

2002-06-01

Full Text Available Females abortion is one of the most important sequela of genital infection with chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum.In this study frequency of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum was studied in 125 females with habitual abortion by direct and indirect immunofluorescence tests and culture method and compared with 250 normal population. The results obtained were as follow: Mycoplasma hominis was isolated from 18 (14.4% females with habitual abortion and 18 (7.2% normal population (P=0.0139. Ureaplasma urealyticum was isolated from 39(31.2% females with habitual aboration and 48 (19.2% normal population (P=0.0045. Chlamydia trachomatis was detected by direct immunofluorescence test in 9 (7.2% of cases and 2 (0.8% of control groups (P=0.0002. the antibody titer against D-K serotypes of Chlamydia trachomatis was also measured. The valuable titer of antibody (>1/16 was detected in 15 (12% of cases and 8 (3.2% of control groups (P=0.0004.The results show that chlamydia trachomatis and Ureaplasma urealyticum may be responsible for some cases of abortion.

Effect of a Mycoplasma hominis-like Mycoplasma on the infection of HEp-2 cells by the TW-183 strain of Chlamydia pneumoniae.

Science.gov (United States)

Castilla, E A; Wadowsky, R M

2000-02-01

We isolated a Mycoplasma hominis-like mycoplasma from a stock culture of Chlamydia pneumoniae TW-183 obtained from the American Type Culture Collection and eradicated the contaminant by treating the stock suspension with a nonionic detergent, Igepal CA-630. The M. hominis-like mycoplasma neither inhibits nor enhances the infectivity of C. pneumoniae for HEp-2 cells.

Effect of a Mycoplasma hominis-Like Mycoplasma on the Infection of HEp-2 Cells by the TW-183 Strain of Chlamydia pneumoniae

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Castilla, Elias A.; Wadowsky, Robert M.

2000-01-01

We isolated a Mycoplasma hominis-like mycoplasma from a stock culture of Chlamydia pneumoniae TW-183 obtained from the American Type Culture Collection and eradicated the contaminant by treating the stock suspension with a nonionic detergent, Igepal CA-630. The M. hominis-like mycoplasma neither inhibits nor enhances the infectivity of C. pneumoniae for HEp-2 cells.

Presencia de Blastocystis Hominis como agente causal de enfermedades gatrointestinales en la comuna 7 (Gaira del Distrito de Santa Marta

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Sonja Liliana Lozano Socarras

2013-10-01

Full Text Available La Blastocystis hominis es un protozoo que causa cuadros diarreicos. Es altamente prevalente en poblaciones que no cuentan con servicios adecuados de higiene, alcantarillado y salud pública. La infección con Blastocystis hominis frecuentemente concomita con otros enteropatógenos de reconocida patogenicidad, además se ha reportado como parásito oportunista en pacientes con VIII SIDA. El objetivo del presente estudio es determinar la presencia de Blastocystis hominis en pacientes de consulta externa con síntomas asociados a enfermedades gastrointestinales, en la comuna 7 del distrito de Santa Marta, Colombia, durante el mes de Enero a Diciembre de 2004. El método de diagnóstico utilizado fue examen coproparasitológico seriado y el número de pacientes analiza-dos fue de 291. Los resultados muestran una alta presencia de Blastocystis hominken pacientes con enfermedad diarreica residentes en Gaira. Un alto porcentaje de la población parasitada (62,6% presento como único agente causal de la enfermedad diarreica al Blastocystis hominis lo que sugiere la presencia de otras enfermedades que pueden involucrar un compromiso inmunológico, el efecto será una respuesta inmune débil contra el parásito.

Selection of Mycoplasma hominis PG21 deletion mutants by cultivation in the presence of monoclonal antibody 552

DEFF Research Database (Denmark)

Jensen, L T; Ladefoged, S; Birkelund, S

1995-01-01

characterized. The mutants showed deletions of a various number of repeats. The deletions were accompanied by a decrease in size of the proteins. With increasing size of deletions, agglutination and growth inhibition by MAb 552 became less pronounced. Spontaneous aggregation of the mutant M. hominis cells...

The Effect of Hominis Placenta Herbal Acupuncture on Bell's palsy

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Yun Jeong-hun

2000-07-01

Full Text Available This report was done to observe the effect of Hominis placenta herbal acupuncture on Bell's palsy. The study group comprised 16 patients who arrived at Woo-suk university oriental hospital from January, 1999 till January, 2000 for Bell's palsy. All patients were divided into two group. One was herbal acupunture group, and the other was control group. Acupunture group was done herbal acupuncture therapy on the facial acupuncture points. Followings are achievement and a term of each group. In herbal acupuncture group, 100% motor recovery was 7 case, 75% was 1 case, and 25% motor recovery term was 7.38±5.21 days, 50% was 11.00±6.16 days, 75% was 15.13±9.55 days, 100% was 23.14±7.97 days. In control group, 100% motor recovery was 4 case, 75% was 2 case, 25% below was 2 case and 25% motor recovery term was 11.17±4.96days, 50% was 18.17±6.82 days, 75% was 29.50±6.95 days, 100% was 44.00±11.49 days. The above results indicate that Hominis placenta herbal acupuncture is a useful effect on Bell's palsy. thus, continuous herbal acupunture study will be needed for more clinical application on Bell' palsy.

Clinical significance of asymptomatic urogenital Mycoplasma hominis and Ureaplasma urealyticum in relation to seminal fluid parameters among infertile Jordanian males

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Hala I. Al-Daghistani

2010-01-01

Conclusion: The differences in the occurrence of M. hominis were statistically insignificant among infertility and control groups, but it was significant for U. urealyticum (p=0.046. M. hominis occurs more frequently in the semen of infertile-varicose male and normal seminal fluid quality. It seems to have no adverse effects on sperm motility but it might decline the fertility potential in such cases. U. urealyticum on the other hand have no clear significant impacts on sperm motility. The mean values for sperm motility, concentrations, and viscosity were not affected by the presence of the two species. Despite the significant presence of Ureaplasma among infertility, further studies were needed to clarify their potential effect on semen quality and infertility status.

Survival and infectivity of Sarcoptes scabiei var. canis and var. hominis.

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Arlian, L G; Runyan, R A; Achar, S; Estes, S A

1984-08-01

Sarcoptes scabiei var. canis served as a suitable model for the study of S. scabiei var. hominis survival. S. scabiei var. canis and var. hominis mites were found to survive off the host for 24 to 36 hours at room conditions (21 degrees C and 40% to 80% relative humidity [RH]), and the canine variety survived 19 days at 10 degrees C and 97% RH. Female mites survived decidedly longer than male mites at comparable conditions. Generally, higher RH values and lower temperatures favored survival, whereas higher temperature and lower RH led to early death. Most canine scabies mites that were held off the host for 36 hours at 75% RH and 22 degrees to 24 degrees C remained infective and penetrated when returned to the host. Live mites of the human variety that were recovered from bed linen slept on by infested patients would also penetrate a host after being held off a host for 96 hours in alternating 12-hour periods of room conditions and refrigeration. Penetration required less than 30 minutes for all life stages of both varieties, and it was accomplished by a mite secretion that dissolved the host tissue. Dislodged mites, particularly those in close proximity to the source, can be a likely source of infestation.

Effect of Hominis placenta Pharmacopuncture on the Dysmenorrhea (A Pilot study, Single blind, Randomized, Controlled Clinical Trial

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Su-Min Kim

2008-09-01

Full Text Available Objective : This study was designed to evaluate the effect of Hominis placenta Pharmacopuncture treatment on Dysmenorrhea of Women. Methods : 49 subjects who were suffering from dysmenorrhea volunteered to answer the MMP(Measure of Menstrual Pain and MSSL(Menstrual Symptom Severity List questionnaire. They were divided into two groups, a Hominis placenta Pharmacopuncture treatment group(Experiment al group, n=25 and a Normal Saline(N/S treatment group(Control group, n=24. The two groups were injected on the CV4, S36, Sp9 and Sp6 acupuncture point. They were treated totally five times depending on the individual menstruation cycles. The scores of MMP and MSSL were measured overall three times before and after the menstruation cycle. The collected data were analyzed as paired t-test, independent t-test using SPSS 12.0 WIN Program. Results : As a result of the evaluation by MMP and MSSL, a significant improvement on dysmenorrhea was made in the two groups(p<0.05, and both scores of Experiment group were decreased more than Control group. But there was no significant difference between the two groups. Conclusions : The Hominis placenta Pharmacopuncture treatment and the Normal Saline treatment were effective in decreasing the symptom of Dysmenorrhea.

Post-embrionic development of the digestive tube of Dermatobia hominis (Linnaeus (Diptera, Cuterebridae

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Ana Maria Vieira

1996-01-01

Full Text Available The digestive tube of 2nd and 3rd instar larvae, pupae and newly emerged adults of Dermatobia hominis (Linnaeus, 1781 was studied anatomically. The specimens were dissected in buffer saline under a stereomicroscope, and the digestive tubes were placed on slides and fixed in 10% buffered formalin. Each tube was measured using a micrometric eye piece, and drawings were made with camera lucida. The results showed that the midgut, the hindgut and the Malpighian tubules with their ducts grow gradually during the larval development. The oesophagus and the salivary glands with their ducts grow only during the moult from the 2nd to the 3rd instar. In the pupal period, salivary glands grow gradually but disappeared after the 20th day. After metamorphosis the digestive tube regressed. This is expected since adult D. hominis lives about nine days without feeding. This fly, similar to other calyptratae muscoid flies shows no vestige of a crop during all post-embrionic development, and the adult has no salivary glands.

Prevalence and characteristics of Blastocystis hominis infection in children.

Science.gov (United States)

O'Gorman, M A; Orenstein, S R; Proujansky, R; Wadowsky, R M; Putnam, P E; Kocoshis, S A

1993-02-01

Blastocystis hominis, a protozoan whose pathogenicity has been questioned, is sometimes found in the human gastrointestinal tract. We sought to determine the prevalence of Blastocystis in stool and to characterize clinical features of infection with Blastocystis in children. Forty-six (3%) of 1,736 patients undergoing fecal microscopy at Children's Hospital of Pittsburgh between January 1, 1985, and December 31, 1988, harbored Blastocystis. Of these 46 children, 75% had exposure to well water or had been in developing countries. Thirty-nine of the 46 (85%) experienced gastrointestinal symptoms, such as abdominal pain, diarrhea, vomiting, and weight loss. Blastocystis was the only parasite found in 35 of those 39 symptomatic children. Symptoms resolved within one month in 90% of patients receiving antiparasitic pharmacotherapy, but in only 58% (P < .04) of those receiving no therapy. We conclude that children infected with Blastocystis often experience gastrointestinal symptoms and that treatment increases the rate of symptomatic improvement. We speculate that Blastocystis is a human pathogen.

Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs.

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Oleaga, Carlota; Bernabini, Catia; Smith, Alec S T; Srinivasan, Balaji; Jackson, Max; McLamb, William; Platt, Vivien; Bridges, Richard; Cai, Yunqing; Santhanam, Navaneetha; Berry, Bonnie; Najjar, Sarah; Akanda, Nesar; Guo, Xiufang; Martin, Candace; Ekman, Gail; Esch, Mandy B; Langer, Jessica; Ouedraogo, Gladys; Cotovio, Jose; Breton, Lionel; Shuler, Michael L; Hickman, James J

2016-02-03

We report on a functional human model to evaluate multi-organ toxicity in a 4-organ system under continuous flow conditions in a serum-free defined medium utilizing a pumpless platform for 14 days. Computer simulations of the platform established flow rates and resultant shear stress within accepted ranges. Viability of the system was demonstrated for 14 days as well as functional activity of cardiac, muscle, neuronal and liver modules. The pharmacological relevance of the integrated modules were evaluated for their response at 7 days to 5 drugs with known side effects after a 48 hour drug treatment regime. The results of all drug treatments were in general agreement with published toxicity results from human and animal data. The presented phenotypic culture model exhibits a multi-organ toxicity response, representing the next generation of in vitro systems, and constitutes a step towards an in vitro "human-on-a-chip" assay for systemic toxicity screening.

Evaluation of First Voided Urine Samples For Detection of Ureaplasma Uriealyticum and Mycoplasma Hominis in Urinary Tracts of Men and Women Suffering from Nongonococcal and Nonspecific Urethritis

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M Mohamadi

2007-07-01

Full Text Available Introduction: Ureaplasma uriealyticum is one of the most important causes of Nongonococcal and Nonspecific urethritis (NGU & NSU in men. Mycoplasma hominis too has a causal role in NGU & NSU. This study aimed to investigate whether it is possible to detect Mycoplasma hominis and Ureaplasma uriealyticum in first voided urine samples in men suffering from NGU & NSU without complaints of urethral secretions and in women with clinical symptoms despite negative vaginal secretion culture test results. Methods: First voided urine samples were taken from 150 patients (21 women & 129 men suffering from NGU & NSU who referred to the Division of Bacteriology, School of Public Health, Tehran University of Medical Sciences in 2004-2005. Samples were examined by culture method. Results: Cultures were positive for Mycoplasma and Ureoplasma in 49 (32.6 % of the 150 samples. Of the 21 samples taken from women, 5 samples were positive for Mycoplasma & Ureoplasma (2 samples Mycoplasma, 3 samples Ureaplasma. Samples from 44 men were positive for Ureoplasma & Mycoplasma(17 samples Mycoplasma, 4 samples Ureaplasma and 23 samples were positive for both. Ureoplasma urealyticum was detected in 30 samples (20% and Mycoplasma hominis, was detected in 42 samples (28%. Conclusion: The results of this study provides evidence that culture tests can be done using voided urine in order to detect Mycoplasma hominis and ureaplasma urealyticum in patients suffering from Nongonococcal urethris; men who do not have urethral secretions and women with clinical symptoms despite negative vaginal secretion culture test results.

PCR-Múltiple para el diagnóstico de Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum y Ureaplasma urealyticum

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Nadia Rodríguez-Preval

2007-04-01

Full Text Available Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum y Ureaplasma urealyticum son especies relacionadas con enfermedades del tracto genitourinario, y particularmente con la uretritis no gonocócica (UNG en el hombre. Los cultivos de estos microorganismos resultan complicados, por lo que las técnicas moleculares, principalmente la reacción en cadena de la polimerasa (PCR, se han convertido en el principal método de detección de estos organismos. Objetivo: Implementar un método molecular basado en tecnología de genes para el diagnóstico de estas cuatro especies de micoplasmas genitales, aplicándolo en muestras clínicas de pacientes con UNG. Material y métodos: Se crearon las condiciones para un PCR-Múltiple para identificar estas especies empleando como muestra ADN de referencia, utilizando los juegos de cebadores complementarios a fragmentos de los genes de la proteína adhesiva de M. genitalium (MgPa, ARN ribosomal 16S de M. hominis, región espaciadora entre los genes del ARN ribosomal 16S y 23S de U. parvum, y de la región espaciadora adyacente al gen de la ureasa y específico para U. urealyticum, siendo un método específico y sensible. Resultados: Al analizar 34 muestras de exudado uretral, 27 correspondieron a la clase Mollicutes, obteniéndose 14,8% de positividad a M. genitalium, 18,5% a M. hominis, 11,1% a U. urealyticum y 3,7%. a U. parvum. Con este trabajo se realizó por primera vez el diagnóstico de M. genitalium, M. hominis, U. parvum y U. urealyticum en muestras uretrales de pacientes cubanos. Conclusión: Se recomienda incluir el diagnóstico de estas especies en un mayor número de pacientes cubanos con síntomas uretrales, para validar el método propuesto y conocer la relación de estos microorganismos con la UNG.

Cryptosporidium hominis Is a Newly Recognized Pathogen in the Arctic Region of Nunavik, Canada: Molecular Characterization of an Outbreak.

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Karine Thivierge

2016-04-01

Full Text Available Cryptosporidium is a leading cause of childhood diarrhea in low-resource settings, and has been repeatedly associated with impaired physical and cognitive development. In May 2013, an outbreak of diarrhea caused by Cryptosporidium hominis was identified in the Arctic region of Nunavik, Quebec. Human cryptosporidiosis transmission was previously unknown in this region, and very few previous studies have reported it elsewhere in the Arctic. We report clinical, molecular, and epidemiologic details of a multi-village Cryptosporidium outbreak in the Canadian Arctic.We investigated the occurrence of cryptosporidiosis using a descriptive study of cases with onset between April 2013 and April 2014. Cases were defined as Nunavik inhabitants of any age presenting with diarrhea of any duration, in whom Cryptosporidium oocysts were detected by stool microscopy in a specialised reference laboratory. Cryptosporidium was identified in stool from 51 of 283 individuals. The overall annual incidence rate (IR was 420 / 100,000 inhabitants. The IR was highest among children aged less than 5 years (1290 /100,000 persons. Genetic subtyping for stool specimens from 14/51 cases was determined by DNA sequence analysis of the 60 kDa glycoprotein (gp60 gene. Sequences aligned with C. hominis subtype Id in all cases. No common food or water source of infection was identified.In this first observed outbreak of human cryptosporidiosis in this Arctic region, the high IR seen is cause for concern about the possible long-term effects on growth and development of children in Inuit communities, who face myriad other challenges such as overcrowding and food-insecurity. The temporal and geographic distribution of cases, as well as the identification of C. hominis subtype Id, suggest anthroponotic rather than zoonotic transmission. Barriers to timely diagnosis delayed the recognition of human cryptosporidiosis in this remote setting.

Update on Pneumocystis carinii f. sp. hominis Typing Based on Nucleotide Sequence Variations in Internal Transcribed Spacer Regions of rRNA Genes

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Lee, Chao-Hung; Helweg-Larsen, Jannik; Tang, Xing; Jin, Shaoling; Li, Baozheng; Bartlett, Marilyn S.; Lu, Jang-Jih; Lundgren, Bettina; Lundgren, Jens D.; Olsson, Mats; Lucas, Sebastian B.; Roux, Patricia; Cargnel, Antonietta; Atzori, Chiara; Matos, Olga; Smith, James W.

1998-01-01

Pneumocystis carinii f. sp. hominis isolates from 207 clinical specimens from nine countries were typed based on nucleotide sequence variations in the internal transcribed spacer regions I and II (ITS1 and ITS2, respectively) of rRNA genes. The number of ITS1 nucleotides has been revised from the previously reported 157 bp to 161 bp. Likewise, the number of ITS2 nucleotides has been changed from 177 to 192 bp. The number of ITS1 sequence types has increased from 2 to 15, and that of ITS2 has increased from 3 to 14. The 15 ITS1 sequence types are designated types A through O, and the 14 ITS2 types are named types a through n. A total of 59 types of P. carinii f. sp. hominis were found in this study. PMID:9508304

Heterogeneity and compartmentalization of Pneumocystis carinii f. sp. hominis genotypes in autopsy lungs

DEFF Research Database (Denmark)

Helweg-Larsen, J; Lundgren, Bettina; Lundgren, Jens Dilling

2001-01-01

The extent and importance of genotype heterogeneity of Pneumocystis carinii f. sp. hominis within lungs have not previously been investigated. Two hundred forty PCR clones obtained from respiratory specimens and lung segments from three patients with fatal P. carinii pneumonia were investigated....... Not all genotypes present in the lungs at autopsy were detected in the diagnostic respiratory samples. Compartmentalization of specific ITS and mtLSU rRNA sequence types was observed in different lung segments. In conclusion, the interpretation of genotype data and in particular ITS sequence types...... in the assessment of epidemiological questions should be cautious since genotyping done on respiratory samples cannot a priori be assumed to represent all genotypes present within the lung....

Co-infection with vaginal Ureaplasma urealyticum and Mycoplasma hominis increases adverse pregnancy outcomes in patients with preterm labor or preterm premature rupture of membranes.

Science.gov (United States)

Kwak, Dong-Wook; Hwang, Han-Sung; Kwon, Ja-Young; Park, Yong-Won; Kim, Young-Han

2014-03-01

The purpose of this study was to determine the prevalence of Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) in patients with preterm labor or preterm premature rupture of membranes (PPROM) and to determine the effect of these organisms on pregnancy outcomes based on the density of colonization. The study group consisted of 184 women with preterm labor or PPROM. Vaginal cultures for UU and MH were performed for all patients at admission, and the placentas were histologically evaluated after delivery. The prevalence of positive vaginal fluid cultures for genital mycoplasma was 62.5% (112/179). This group included 99 patients carrying only UU and 13 carrying both organisms. No patients were found to carry only MH. Compared to patients only positive for UU, patients with both organisms showed significantly decreased gestational age at birth and birth weight, and significant increases in the incidences of preterm birth, NICU admissions and histologic chorioamnionitis. Vaginal MH tends to be detected with UU, and patients carrying both organisms simultaneously had more severe adverse pregnancy outcomes compared to patients in preterm labor or PPROM who were only positive for UU.

Frequency of Blastocystis hominis and other intestinal parasites in stool samples examined at the Parasitology Laboratory of the School of Pharmaceutical Sciences at the São Paulo State University, Araraquara Freqüência de Blastocystis hominis e outros enteroparasitas em amostras de fezes examinadas no Laboratório de Parasitologia da Faculdade de Ciências Farmacêuticas da Universidade Estadual Paulista, Araraquara

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Júlio César Miné

2008-12-01

Full Text Available Blastocystis homins is a protozoan that causes an intestinal infection known as human blastocystosis. This infection is diagnosed by means of parasitological examination of stools and by permanent staining techniques. The present study was developed to evaluate the frequency of Blastocystis hominis infection among inhabitants of the Araraquara region, State of São Paulo, and to compare different methods for investigating this protozoan in feces samples. Evaluations on 503 stool samples were performed by means of direct fresh examination and using the techniques of Faust et al., Lutz and Rugai et al. In addition, the iron hematoxylin, trichrome and modified Kinyoun staining techniques were used. Out of the 503 samples examined, 174 (34.6% were found to be positive for the presence of intestinal parasites. The most frequent protozoa and helminths were Entamoeba coli (14.6% and Strongyloides stercoralis (6.7%, respectively. Blastocystis hominis was present in 23 (4.6% fecal samples, with a predominately pasty consistency and without characterizing a condition of diarrhea. Despite the low frequency of Blastocystis hominis found in the Araraquara region, compared with other regions of Brazil, it is important to perform laboratory diagnostic tests for this protozoan. Its finding in fecal material is indicative of food and drinking water contamination. Since the transmission route for this parasite is accepted to be oral-fecal, this implies that the population needs guidance regarding hygiene and basic sanitation measures as a means for controlling health problems caused by enteroparasites.Blastocystis hominis é um protozoário, causador de infecção intestinal denominada blastocistose humana, cujo diagnóstico é realizado pelo exame coproparasitológico e por meio de técnicas de coloração permanente. Este estudo foi desenvolvido para avaliar a freqüência da infecção por Blastocystis hominis em habitantes da região de Araraquara/SP, bem

First report of Sneathia sanguinegens together with Mycoplasma hominis in postpartum prosthetic valve infective endocarditis: a case report.

Science.gov (United States)

Kotaskova, Iva; Nemec, Petr; Vanerkova, Martina; Malisova, Barbora; Tejkalova, Renata; Orban, Marek; Zampachova, Vita; Freiberger, Tomas

2017-08-14

The presence of more than one bacterial agent is relatively rare in infective endocarditis, although more common in prosthetic cases. Molecular diagnosis from a removed heart tissue is considered a quick and effective way to diagnose fastidious or intracellular agents. Here we describe the case of postpartum polymicrobial prosthetic valve endocarditis in a young woman. Sneathia sanguinegens and Mycoplasma hominis were simultaneously detected from the heart valve sample using broad range 16S rRNA polymerase chain reaction (PCR) followed by sequencing while culture remained negative. Results were confirmed by independent PCR combined with denaturing gradient gel electrophoresis. Before the final agent identification, the highly non-compliant patient left from the hospital against medical advice on empirical intravenous treatment with aminopenicillins, clavulanate and gentamicin switched to oral amoxycillin and clavulanate. Four months after surgery, no signs of inflammation were present despite new regurgitation and valve leaflet flail was detected. However, after another 5 months the patient died from sepsis and recurrent infective endocarditis of unclarified etiology. Mycoplasma hominis is a rare causative agent of infective endocarditis. To the best of our knowledge, presented case is the first report of Sneathia sanguinegens detected in this condition. Molecular techniques were shown to be useful even in polymicrobial infective endocarditis samples.

Mycoplasma hominis Symbiosis and Trichomonas vaginalis Metronidazole Resistance%人型支原体共生与阴道毛滴虫甲硝唑耐药性的关系

Institute of Scientific and Technical Information of China (English)

王频佳; 谢成彬

2012-01-01

目的 研究人型支原体(Mycoplasma hominis)的共生与阴道毛滴虫(Trichomonas vaginalis)甲硝唑耐药性的关系.方法 2010年11月至2011年7月,自四川省妇幼保健院妇科门诊患者生殖道分泌物中分离出160株阴道毛滴虫,用梯度浓度1 024、512、256……4、2和1 μg/ml甲硝唑分别处理该批虫株,以死亡率≥90％的最低浓度作为甲硝唑最小致死浓度(MLC).以160个阴道毛滴虫分离株中提取的DNA为模板,用PCR技术特异性扩增人型支原体16S rRNA基因,检测滴虫细胞内是否有人型支原体共生.对检出人型支原体DNA的分离株用32μg/ml多西环素清除支原体,比较清除前后甲硝唑MLC的变化.结果 160个阴道毛滴虫分离株中甲硝唑MLC为1～8μg/ml的占61.3％(98/160),16～32μg/ml的占26.3％(42/160),64～256μg/ml的占12.5％(20/160).PCR检测结果显示,有61株(38.1％)检出人型支原体DNA,其中MLC为1～8μg/ml的分离株检出率为13.3％(13/98),16～32μg/ml的分离株检出率为73.8％( 31/42),64～256μg/ml的分离株检出率为85.0％(17/20),不同MLC范围的分离株人型支原体检出率差异有统计学意义(P＜0.01).用多西环素处理后,61株中仅有8株支原体被清除,清除前后甲硝唑MLC无明显变化.结论 四川地区的阴道毛滴虫分离株对甲硝唑表现出一定程度的耐药性,人型支原体的共生可能与之有关,但尚未发现直接证据.%Objective To investigate the relation of Mycoplasma hominis symbiosis and the resistence of Trichomonas vaginalit to metronidazole. Methods From November 2010 to July 2011, 160 isolates of T. Vaginalis were collected from the genital tract secretion of gynecological out-patients at the Sichuan Provincial Hospital for Women and Children. The minimum lethal concentration (MLC) to metronidazole of these isolates was determined by an in. Vitro sensitivity assay with different concentration gradients of metronidazole (from 1 to 1 024 μg/ml), and M

A Clinical Study on 1 Case of Patient with Bilateral Simultaneous Bell's Palsy Treated by Hominis Placenta Herbal-Acupuncture

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Kwon, Kang

2003-06-01

Full Text Available Objective : This study was carried out to investigate the progress of bilateral simultaneous facial palsy and the effect of Hominis Placenta herbal-acupunture and the other oriental medical therapies. Methods : We used two methods to research the progress of disease. 1. Diagnosis - Facial muscle test, Taste test, Hearing test, Photographies, Lab-finding 2. Treatment - Acupuncture, Herbal-acupuncture, Electroacupuncture, Herb-med Results : The onset of Rt. facial palsy was earlier than Lt. facial palsy 3days. The reaction on the treatment of Rt. facial palsy was more dull than Lt. facial palsy. In terms of treatment period, Rt. facial palsy was very longer than Lt. facial palsy. Conclusion : According to the above results, we discoveried that Hominis Placenta herbal-acupunture and the other oriental medical therapies had good influence on the bilateral simultaneous facial palsy. In the future, we should endeavor to know influence between Rt. and Lt. face in case of bilateral simultaneous Bell's palsy.

ヒト精子運動に及ぼす Ureaplasma urealyticum 及び Mycoplasma hominis の影響

OpenAIRE

Chang, Myung Woong; Choi, Tae Kyung; Matsuo, Yoshiyasu; Yoshii, Zensaku

1984-01-01

The human spermatozoal motility was significantly affected by the whole culture solution and slightly less significantly by the microbial cell suspension of Ureaplasma urealyticum but not the microbial metabolites (culture supernatant). The same phenomenon was observed with the preparations of Mycoplasma hominis in somewhat less degree.

Cytokine concentrations in seminal plasma from subfertile men are not indicative of the presence of Ureaplasma urealyticum or Mycoplasma hominis in the lower genital tract

NARCIS (Netherlands)

Pannekoek, Y.; Trum, J. W.; Bleker, O. P.; van der Veen, F.; Spanjaard, L.; Dankert, J.

2000-01-01

The inflammatory response to the presence of Ureaplasma urealyticum or Mycoplasma hominis in the lower genital tract of subfertile men without any signs or symptoms of infection was investigated by measuring the concentrations of interleukin (IL)-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and

IN VITRO PHYTOREMEDIATION OF PERSISTENT ORGANIC POLLUTANTS BY Helianthus annuus L. PLANTS

OpenAIRE

Almeida, Marcos V. de; Rissato, Sandra R.; Galhiane, Mário S.; Fernandes, João R.; Lodi, Paulo C.; Campos, Marcelo C. de

2018-01-01

Plant model systems are needed to properly conduct basic laboratory studies prior to field applications of phytoremediation. In vitro plant cultures are a useful tool for such research. This study focuses on the removal and/or degradation of 24 persistent organic pollutants under in vitro conditions by Helianthus annuus L (sunflower). The main purpose of exploiting this plant for phytoremediation process is due to its strong adaptability to adverse environments conditions such as resistance t...

Detection of T. vaginalis,M. hominis,M. genitalium, C. trachomatis, N. gonorrhoeae and U. urealyticum using Multiplex PCR

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Tamara Brunelli

2013-03-01

Full Text Available Intoduction. The sexually transmitted diseases include a large group of infections affecting both the sexes. In this study we evaluated the prevalence of Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma genitalium, Chlamydia trachomatis, Neisseria gonorrhoeae and Ureaplasma urealyticum in the Prato area during the period September 2010 – July 2011. Methods.We analysed different kind of samples (urine, endocervical swabs, urethral swabs, seminal fluids from hospitalized patients or referred to the Prato clinic subjects.The DNA was obtained using EZ1-DNA extraction kit and EZ1 instrument.The DNA was then amplified using the Seeplex STD6 kit (Seegene, Korea, identifying multiple pathogens simultaneously (T. vaginalis, M. hominis, M. genitalium, C. trachomatis, N. gonorrhoeae e U. urealyticum. The revelation was performed by electrophoresis on microchip (instrument Multina, Shimadzu, Japan. Results. 1136 samples from Italian and foreign patients were examined: 876 were endocervical swabs (77%, 103 urethral swabs (9%, 103 seminal fluids (9%, and 54 urines (5%. The number of females was higher than males [894 (78.7% vs 242 (21.3%]; the mean age of females was 37.0±11.6 years, whereas that of males was 41.5 ±12.63 years.The prevalence of urogenital pathogens was: 15 positive samples for T. vaginalis (1.3%, 56 for M. hominis (4.9%, 13 for M. genitalium (1.1%, 28 for C. trachomatis (2.5%, 8 for N. gonorrhoeae (0.7% and 87 for U. urealyticum (7.7%.Among all positive, 25 subjects were positive for more than one pathogen and in particular: one was positive for the presence of 4 pathogens, five presented 3 pathogens simultaneously and the remaining nineteen for 2 pathogens. Conclusions. This study provides data on the prevalence of sexually transmitted diseases in the hospital of Prato.

Antibacterial Resistance in Ureaplasma Species and Mycoplasma hominis Isolates from Urine Cultures in College-Aged Females

Science.gov (United States)

Valentine-King, Marissa A.

2017-01-01

ABSTRACT Urinary tract infections (UTIs) affect nearly 20% of women age 15 to 29 and account for an estimated $3.5 billion in costs. Antibiotic resistance prolongs UTI treatment, and resistance profiles vary regionally. This regional variation is an important consideration in guiding empirical treatment selection. Regional studies in the United States have identified tetracycline resistance in over one-third of Ureaplasma species isolates, but no studies have evaluated antibiotic resistance levels in college-aged women with a first-time UTI. We tested a panel of antibiotics and determined the MICs of Ureaplasma species (60 U. parvum and 13 U. urealyticum) and 10 Mycoplasma hominis isolates obtained from urine from college-aged women with a first-time UTI. Low antibiotic resistance was found in this population of women with a first-time UTI. All M. hominis and U. urealyticum isolates were sensitive. However, two U. parvum isolates were resistant, with one to levofloxacin (MIC, 4 μg/ml) and one to tetracycline (MIC, 8 μg/ml). For the Ureaplasma spp., the MIC90s were highest against gentamicin (21 μg/ml) and lowest against doxycycline (0.25 μg/ml). In a comparison of MIC levels between Ureaplasma spp., U. urealyticum had significantly higher MICs against each antibiotic except doxycycline. For the resistant isolates, the genetic mechanisms of resistance were determined. PCR amplification identified tetM to be present in the tetracycline-resistant isolate and an S83W mutation within the parC gene of the quinolone-resistant isolate. To our knowledge, this study is the first to provide molecular and phenotypic evidence of the S83W parC mutation conferring levofloxacin resistance in U. parvum isolated from a patient in the United States. PMID:28827422

Allometric scaling and cell ratios in multi-organ in vitro models of human metabolism

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Nadia eUcciferri

2014-12-01

Full Text Available Intelligent in vitro models able to recapitulate the physiological interactions between tissues in the body have enormous potential as they enable detailed studies on specific two-way or higher order tissue communication. These models are the first step towards building an integrated picture of systemic metabolism and signalling in physiological or pathological conditions. However the rational design of in vitro models of cell-cell or cell-tissue interaction is difficult as quite often cell culture experiments are driven by the device used, rather than by design considerations. Indeed very little research has been carried out on in vitro models of metabolism connecting different cell or tissue types in a physiologically and metabolically relevant manner. Here we analyse the physiologic relationship between cells, cell metabolism and exchange in the human body using allometric rules, downscaling them to an organ-on-a plate device. In particular, in order to establish appropriate cell ratios in the system in a rational manner, two different allometric scaling models (Cell Number Scaling Model, CNSM, and Metabolic and Surface Scaling model, MSSM are proposed and applied to a two compartment model of hepatic-vascular metabolic cross-talk. The theoretical scaling studies illustrate that the design and hence relevance of multi-organ models is principally determined by experimental constraints. Two experimentally feasible model configurations are then implemented in a multi-compartment organ-on-a plate device. An analysis of the metabolic response of the two configurations demonstrates that their glucose and lipid balance is quite different, with only one of the two models recapitulating physiological-like homeostasis. In conclusion, not only do cross-talk and physical stimuli play an important role in in vitro models, but the numeric relationship between cells is also crucial to recreate in vitro interactions which can be extrapolated to the in vivo

Allometric Scaling and Cell Ratios in Multi-Organ in vitro Models of Human Metabolism

International Nuclear Information System (INIS)

Ucciferri, Nadia; Sbrana, Tommaso; Ahluwalia, Arti

2014-01-01

Intelligent in vitro models able to recapitulate the physiological interactions between tissues in the body have enormous potential as they enable detailed studies on specific two-way or higher order tissue communication. These models are the first step toward building an integrated picture of systemic metabolism and signaling in physiological or pathological conditions. However, the rational design of in vitro models of cell–cell or cell–tissue interaction is difficult as quite often cell culture experiments are driven by the device used, rather than by design considerations. Indeed, very little research has been carried out on in vitro models of metabolism connecting different cell or tissue types in a physiologically and metabolically relevant manner. Here, we analyze the physiological relationship between cells, cell metabolism, and exchange in the human body using allometric rules, downscaling them to an organ-on-a-plate device. In particular, in order to establish appropriate cell ratios in the system in a rational manner, two different allometric scaling models (cell number scaling model and metabolic and surface scaling model) are proposed and applied to a two compartment model of hepatic-vascular metabolic cross-talk. The theoretical scaling studies illustrate that the design and hence relevance of multi-organ models is principally determined by experimental constraints. Two experimentally feasible model configurations are then implemented in a multi-compartment organ-on-a-plate device. An analysis of the metabolic response of the two configurations demonstrates that their glucose and lipid balance is quite different, with only one of the two models recapitulating physiological-like homeostasis. In conclusion, not only do cross-talk and physical stimuli play an important role in in vitro models, but the numeric relationship between cells is also crucial to recreate in vitro interactions, which can be extrapolated to the in vivo reality.

Allometric Scaling and Cell Ratios in Multi-Organ in vitro Models of Human Metabolism

Energy Technology Data Exchange (ETDEWEB)

Ucciferri, Nadia [CNR Institute of Clinical Physiology, Pisa (Italy); Interdepartmental Research Center “E. Piaggio”, University of Pisa, Pisa (Italy); Sbrana, Tommaso [Interdepartmental Research Center “E. Piaggio”, University of Pisa, Pisa (Italy); Ahluwalia, Arti, E-mail: arti.ahluwalia@unipi.it [CNR Institute of Clinical Physiology, Pisa (Italy); Interdepartmental Research Center “E. Piaggio”, University of Pisa, Pisa (Italy)

2014-12-17

Intelligent in vitro models able to recapitulate the physiological interactions between tissues in the body have enormous potential as they enable detailed studies on specific two-way or higher order tissue communication. These models are the first step toward building an integrated picture of systemic metabolism and signaling in physiological or pathological conditions. However, the rational design of in vitro models of cell–cell or cell–tissue interaction is difficult as quite often cell culture experiments are driven by the device used, rather than by design considerations. Indeed, very little research has been carried out on in vitro models of metabolism connecting different cell or tissue types in a physiologically and metabolically relevant manner. Here, we analyze the physiological relationship between cells, cell metabolism, and exchange in the human body using allometric rules, downscaling them to an organ-on-a-plate device. In particular, in order to establish appropriate cell ratios in the system in a rational manner, two different allometric scaling models (cell number scaling model and metabolic and surface scaling model) are proposed and applied to a two compartment model of hepatic-vascular metabolic cross-talk. The theoretical scaling studies illustrate that the design and hence relevance of multi-organ models is principally determined by experimental constraints. Two experimentally feasible model configurations are then implemented in a multi-compartment organ-on-a-plate device. An analysis of the metabolic response of the two configurations demonstrates that their glucose and lipid balance is quite different, with only one of the two models recapitulating physiological-like homeostasis. In conclusion, not only do cross-talk and physical stimuli play an important role in in vitro models, but the numeric relationship between cells is also crucial to recreate in vitro interactions, which can be extrapolated to the in vivo reality.

Allometric Scaling and Cell Ratios in Multi-Organ in vitro Models of Human Metabolism.

Science.gov (United States)

Ucciferri, Nadia; Sbrana, Tommaso; Ahluwalia, Arti

2014-01-01

Intelligent in vitro models able to recapitulate the physiological interactions between tissues in the body have enormous potential as they enable detailed studies on specific two-way or higher order tissue communication. These models are the first step toward building an integrated picture of systemic metabolism and signaling in physiological or pathological conditions. However, the rational design of in vitro models of cell-cell or cell-tissue interaction is difficult as quite often cell culture experiments are driven by the device used, rather than by design considerations. Indeed, very little research has been carried out on in vitro models of metabolism connecting different cell or tissue types in a physiologically and metabolically relevant manner. Here, we analyze the physiological relationship between cells, cell metabolism, and exchange in the human body using allometric rules, downscaling them to an organ-on-a-plate device. In particular, in order to establish appropriate cell ratios in the system in a rational manner, two different allometric scaling models (cell number scaling model and metabolic and surface scaling model) are proposed and applied to a two compartment model of hepatic-vascular metabolic cross-talk. The theoretical scaling studies illustrate that the design and hence relevance of multi-organ models is principally determined by experimental constraints. Two experimentally feasible model configurations are then implemented in a multi-compartment organ-on-a-plate device. An analysis of the metabolic response of the two configurations demonstrates that their glucose and lipid balance is quite different, with only one of the two models recapitulating physiological-like homeostasis. In conclusion, not only do cross-talk and physical stimuli play an important role in in vitro models, but the numeric relationship between cells is also crucial to recreate in vitro interactions, which can be extrapolated to the in vivo reality.

Antibiotic susceptibility profiles of Mycoplasma hominis and Ureaplasma urealyticum isolated during a population-based study concerning women infertility in northeast Romania

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Mareş Mihai

2011-03-01

Full Text Available The study was carried out on 1068 infertile women under initial evaluation. For Mycoplasma hominis, the highest resistance rates were registered for ciprofloxacin (72.22%, followed by macrolides and ofloxacin. For Ureaplasma urealyticum, the ciprofloxacin resistance was also high (51.72%, while the resistance rates to other tested antibiotics were significantly lower.

Prevalence of the protozoan parasite Cryptosporidium on three organic pig farms in Denmark

DEFF Research Database (Denmark)

Petersen, Heidi H.; Jianmin, Wang; Mejer, Helena

2013-01-01

Pigs are a potential source of contamination with Cryptosporidium spp., which can lead to infection in humans. Two species C. parvum and C. hominis can cause an acute diarrheal illness in humans, which can become severe in e.g. patients with HIV. The oocyst can survive for long periods in the env......Pigs are a potential source of contamination with Cryptosporidium spp., which can lead to infection in humans. Two species C. parvum and C. hominis can cause an acute diarrheal illness in humans, which can become severe in e.g. patients with HIV. The oocyst can survive for long periods...... in the environment and is resistant to many common disinfectants. In order to estimate the prevalence of the zoonotic parasite Cryptosporidium in organic pigs and to improve our knowledge of the parasite epidemiology, the prevalence was monitored four times between September 2011 and June 2012 in three Danish...... organic pig farms. Faecal samples were collected for examination of Cryptosporidium spp. with a total of 994 pigs grouped as sows, fatteners, young pigs and piglets. The number of pigs in each age group was 298, 232, 315 and 161 respectively, distributed on the three farms. Faecal samples were collected...

DNA sequencing reveals limited heterogeneity in the 16S rRNA gene from the rrnB operon among five Mycoplasma hominis isolates

DEFF Research Database (Denmark)

Mygind, T; Birkelund, Svend; Christiansen, Gunna

1998-01-01

To investigate the intraspecies heterogeneity within the 16S rRNA gene of Mycoplasma hominis, five isolates with diverse antigenic profiles, variable/identical P120 hypervariable domains, and different 16S rRNA gene RFLP patterns were analysed. The 16S rRNA gene from the rrnB operon was amplified...

Organs-on-a-chip: Current applications and consideration points for in vitro ADME-Tox studies.

Science.gov (United States)

Ishida, Seiichi

2018-02-01

Assay systems using in vitro cultured cells are increasingly applied for evaluation of the efficacy, safety, and toxicity of drug candidates. In vitro cell-based assays have two main applications in the drug discovery process: searching for a compound that is effective against the target disease (seed investigation) and confirmation of safety during use of the identified compounds (safety assessment). Currently available in vitro cell-based assays have been designed to evaluate the efficacy and toxicity in single organs, but the in vivo pharmacokinetics and pharmacodynamics of the administered drug candidates have not been considered. Thus, an evaluation system that interconnects cell culture units, one of which has appropriate drug metabolism activities and the other assesses the efficacy and toxicity of compounds, is needed. Accordingly, the in vitro ADME-Tox culture system known as organs-on-a-chip has been proposed. In this review, after introducing the organs-on-a-chip system, the evaluation of enterohepatic circulation and the gut-liver axis relationship will be presented as an example of the application of the organs-on-a-chip system for ADME studies based on inter-organ network. Additionally, the functions required for the organs-on-a-chip system and the necessity of standardization of cells mounted on the chip system will be discussed. Copyright © 2018 The Japanese Society for the Study of Xenobiotics. Published by Elsevier Ltd. All rights reserved.

THE METHODS OF LABORATORY DIAGNOSTICS OF UROGENITAL INFECTIONS ASSOCIATED WITH MYCOPLASMA HOMINIS AND UREAPLASMA SPP.

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O. V. Zarucheynova

2014-01-01

Full Text Available Wide distribution of urogenital mycoplasmas in the population, the high frequency of carrier state and a long asymptomatic course of disease, the lack of specific clinical symptoms making the diagnosis impossible without using of special laboratory tests. The review focuses on indications for mycoplasma infection screening and for an appointmentof antibiotic therapy. The most commonly used laboratory diagnostic methods of urogenital infections, associated with Mycoplasma hominis and Ureaplasma spp., with their characteristics, advantages and disadvantages are described.

Micoplasma hominis, Ureaplasma urealyticum y bacterias aeróbicas en el semen de hombres que consultan por infertilidad Micoplasma hominis, Ureaplasma urealyticum and aerobic bacteria present in the semen from men attending infertility service

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Bertha Victoria Rodríguez Pendás

2013-04-01

.Introduction: human semen infections can alter the sperm quality and be associated to male infertility disorders. Objectives: to determine the frequency of infections from Micoplasma hominis, Ureaplasma urealyticum and other aerobic bacteria in the semen of men who attended the infertility service, and to identify whether there is some relation between the detected infections and the altered semen quality variables or not. Methods: a cross-sectional descriptive study was performed to evaluate semen samples from 140 men aged 20 to 45 years, who attended the infertility service at the National Institute of Endocrinology. According to the WHO guidelines, a complete spermiogram including leukocytospermia was performed in order to determine the qualitative and quantitative variables in the semen. The semen samples were cultured in blood agar and in chocolate agar at 37oC under CO2 environment to find out possible aerobic bacteria. To this end, a set of reagents known as Mycoplasma System Plus was used, allowing the culture, the identification, the semi-quantitative count and the antibiogram of urogenital mycoplasms/ureaplasms. The ethical aspects were allowed for; the results were analyzed through percentage estimations and the chi square test. Results: out of the 140 evaluated semen samples, 58 (41.4 % showed some infection, 37 of them were caused by Ureaplasma urealyticum (25.7 %, 2 by Micoplasma hominis (1.4 % and 19 by the aerobic bacteria (13.8 %. When making a comparison of the qualitative and quantitative variables of the semen from infected and non-infected subjects, there were not any statistically significant differences in the evaluated variables of the sperm quality. Conclusions: the total frequency of infections in the studied sample was relatively high, but was not associated to altered seminal variables.

In vitro organic matter digestibility and gas production of fish-meal ...

African Journals Online (AJOL)

In this study, an in vitro rumen gas production technique was utilized to evaluate fish-meal coated with different types and levels of fats for total gas production, Metabolizable energy (ME) and organic matter digestibility (OMD) contents. Approximately 200 mg of sample was weighed and inserted in glass syringes, then ...

Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies.

Science.gov (United States)

Lemos, Vanessa; Graczyk, Thaddeus K; Alves, Margarida; Lobo, Maria Luísa; Sousa, Maria C; Antunes, Francisco; Matos, Olga

2005-12-01

In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples.

Comparison of Assays for Sensitive and Reproducible Detection of Cell Culture-Infectious Cryptosporidium parvum and Cryptosporidium hominis in Drinking Water

Science.gov (United States)

Di Giovanni, George D.; Rochelle, Paul A.

2012-01-01

This study compared the three most commonly used assays for detecting Cryptosporidium sp. infections in cell culture: immunofluorescent antibody and microscopy assay (IFA), PCR targeting Cryptosporidium sp.-specific DNA, and reverse transcriptase PCR (RT-PCR) targeting Cryptosporidium sp.-specific mRNA. Monolayers of HCT-8 cells, grown in 8-well chamber slides or 96-well plates, were inoculated with a variety of viable and inactivated oocysts to assess assay performance. All assays detected infection with low doses of flow cytometry-enumerated Cryptosporidium parvum oocysts, including infection with one oocyst and three oocysts. All methods also detected infection with Cryptosporidium hominis. The RT-PCR assay, IFA, and PCR assay detected infection in 23%, 25%, and 51% of monolayers inoculated with three C. parvum oocysts and 10%, 9%, and 16% of monolayers inoculated with one oocyst, respectively. The PCR assay was the most sensitive, but it had the highest frequency of false positives with mock-infected cells and inactivated oocysts. IFA was the only infection detection assay that did not produce false positives with mock-infected monolayers. IFA was also the only assay that detected infections in all experiments with spiked oocysts recovered from Envirochek capsules following filtration of 1,000 liters of treated water. Consequently, cell culture with IFA detection is the most appropriate method for routine and sensitive detection of infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water. PMID:22038611

滴虫阴道炎合并人型支原体感染的临床用药初探

Institute of Scientific and Technical Information of China (English)

罗翠连; 陈怡红; 刘英华

2012-01-01

Trichomonas vaginalis and Mycoplasma hominis is a common parasitic in the female urogenital tract pathogens,which can cause genital tract inflammation.In vitro experiments confirmed the two into a symbiotic relationship and the resistance,thereby causing trichomoniasis vaginitis is difficult to cure.This study using molecular biology technique and clinical testing drug sensitivity test,analysis of trichomonas vaginitis with Mycoplasma hominis infection of Mycoplasma hominis resistance,so as to guide the clinical treatment drug of choice.

External ophthalmomyiasis due to Dermatobia hominis. A case report.

Science.gov (United States)

Serra Moltó, A; Molina Martín, J C; Mengual Verdú, E; Hueso Abancens, J R

2018-03-23

A 46-year-old woman with no relevant medical history, native of Honduras and resident in Spain for one and a half months. The patient went to the Emergency Department due to inflammation of the upper eyelid of the right eye, with an area that simulated an abscess. This was drained (obtaining hardly any purulent content). Treatment was prescribed with oral and topical antibiotics, as well as an anti-inflammatory drug. One week later the patient returned, with improvement of the inflammatory signs, but with discomfort and corneal erosions. After eversion of the upper eyelid, a «worm» type parasite emerged from the tarsus. The extraction was completed with a clamp, and was later identified as Dermatobia hominis (Dh) by examination of a fresh specimen. The subsequent outcome of the patient was favourable. Preseptal cellulitis in patients from tropical and sub-tropical areas can be caused by Dh. Copyright © 2018 Sociedad Española de Oftalmología. Publicado por Elsevier España, S.L.U. All rights reserved.

Association between preterm labor and genitourinary tract infections caused by Trichomonas vaginalis, Mycoplasma hominis, Gram-negative bacilli, and coryneforms

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Alaa El-Dien M.S. Hosny

2017-09-01

Conclusion: Our study demonstrated that the main risk factors for PTL were vaginal infection with T. vaginalis, M. hominis, coryneforms, and Gram-negative bacilli, and their determinants (vaginal pH>5, positive whiff test, heavy vaginal bleeding. Both young age (< 20 years and poor obstetric history were also the risk factors. Therefore, screening for genitourinary tract infections is strongly recommended to be included in prenatal care.

Molecular characterization of Cryptosporidium parvum and Cryptosporidium hominis GP60 subtypes worldwide

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Catalina Avendaño V

2017-09-01

Full Text Available Cryptosporidium is a zoonotic parasite very important in animal health as well as in public health. It is because this is one of the main causes of diarrhea in children, calves, lambs and other variety of youth mammalians in a lot of countries. The globalization has enabled the exchange of biological material in different regions worldwide, encouraging the spread of diseases and exposure to these biological agents to different environmental conditions, inducing adaptation through genetic changes. Based in the polymorphism of the gene for GP60, this review intended to present the distribution of Cryptosporidium parvum and Cryptosporidium hominis in humans and calves worldwide. The subtype that affects cattle more frequently corresponds to IIaA15G2R; while the subtype most frequently isolated from human samples is IaA19G2.

Blastocystis Hominis and Chronic Abdominal Pain in Children: Is there an Association between Them?

Science.gov (United States)

Toro Monjaraz, Erick Manuel; Vichido Luna, Miguel Angel; Montijo Barrios, Ericka; Cervantes Bustamante, Roberto; Zárate Mondragón, Flora; Huante Anaya, Alfonso; Cadena León, José; Mendez, Monserrat Cazares; López Ugalde, Martha; Ramirez Mayans, Jaime A

2017-08-03

Chronic abdominal pain has many etiologies, one of them being parasites. The aim of this study was to find an association between chronic abdominal pain in children and Blastocystis hominis (Bh). Clinical files of patients with Bh and functional abdominal pain were reviewed. A comparison was made between patients who showed an improvement of their symptoms and those who did not. Out of the 138 patients who had functional abdominal pain and Bh, 37 patients did not receive any treatment (26.8%), while 101 received it and were treated with different antimicrobial agents (73.2%); regarding the improvement of symptoms, a statistically significant difference (p abdominal pain in children has different etiologies; however, we have documented through this work that it is appropriate to provide antimicrobial treatment for patients with Bh and chronic abdominal pain. © The Author [2017]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Investigation on Symbiosis of Mycoplasma hominis in Trichomonas vaginalis in Some Areas of Guizhou Province%贵州省部分地区阴道毛滴虫与人型支原体共生情况

Institute of Scientific and Technical Information of China (English)

匡贵榕; 衣凤芸; 杨宇箭; 程其会; 王丽娟; 吴家红

2015-01-01

目的：了解贵州地区阴道毛滴虫临床分离株与人型支原体共生情况。方法：以贵州部分地区女性阴道毛滴虫病患者阴道后穹隆分泌物中采集的阴道毛滴虫虫株，实验室达到纯培养后，采用Chelex-100的方法提取滴虫基因组DNA，设计人型支原体16S rDNA特异引物，进行聚合酶链式反应（ PCR），琼脂糖凝胶电泳法对PCR产物进行分析，观察滴虫细胞内人型支原体共生情况。结果：从临床共采集到165株虫株中，83株获得纯培养；83株中人型支原体检测结果显示有47株阳性，阳性率为56.6%。结论：阴道毛滴虫虫株与人型支原体共生情况在我国贵州地区普遍性存在。%Objective:To investigate the symbiosis of Mycoplasma hominis in Trichomonas vaginalis in Guizhou Province. Methods:The clinical strains of T. vaginalis were collected from the hospitals and family planning service stations in Guiyang city and Anshun city. After pure culture in lab,the ge-nome DNA of T. vaginalis was extracted by Chelex-100,and 16S rDNA fragment of M. hominis was amplified by PCR with the specific primers. Agarose gel electrophoresis analysis was conducted and the symbiosis of M. hominis in T. vaginalis was observed. Results:A total of 165 strains were collected from the hospitals and family planning service stations,and 83 clinical strains attained pure culture,of which 47 strains were detected positively for M. hominis. The positive rate was 56. 6%. Conclusions:The symbiosis of M. hominis in T. vaginalis is prevalent in Guizhou province.

Genomic resources and draft assemblies of the human and porcine varieties of scabies mites, Sarcoptes scabiei var. hominis and var. suis.

Science.gov (United States)

Mofiz, Ehtesham; Holt, Deborah C; Seemann, Torsten; Currie, Bart J; Fischer, Katja; Papenfuss, Anthony T

2016-06-02

The scabies mite, Sarcoptes scabiei, is a parasitic arachnid and cause of the infectious skin disease scabies in humans and mange in other animal species. Scabies infections are a major health problem, particularly in remote Indigenous communities in Australia, where secondary group A streptococcal and Staphylococcus aureus infections of scabies sores are thought to drive the high rate of rheumatic heart disease and chronic kidney disease. We sequenced the genome of two samples of Sarcoptes scabiei var. hominis obtained from unrelated patients with crusted scabies located in different parts of northern Australia using the Illumina HiSeq. We also sequenced samples of Sarcoptes scabiei var. suis from a pig model. Because of the small size of the scabies mite, these data are derived from pools of thousands of mites and are metagenomic, including host and microbiome DNA. We performed cleaning and de novo assembly and present Sarcoptes scabiei var. hominis and var. suis draft reference genomes. We have constructed a preliminary annotation of this reference comprising 13,226 putative coding sequences based on sequence similarity to known proteins. We have developed extensive genomic resources for the scabies mite, including reference genomes and a preliminary annotation.

Sub-chronic toxicity of low concentrations of industrial volatile organic pollutants in vitro

International Nuclear Information System (INIS)

McDermott, Catherine; Allshire, Ashley; Pelt, Frank N.A.M. van; Heffron, James J.A.

2007-01-01

Organic solvents form an important class of pollutants in the ambient air and have been associated with neurotoxicity and immunotoxicity in humans. Here we investigated the biological effects of sub-chronic exposure to industrially important volatile organic solvents in vitro. Jurkat T cells were exposed to toluene, n-hexane and methyl ethyl ketone (MEK) individually for 5 days and solvent exposure levels were confirmed by headspace gas chromatography. A neuroblastoma cell line (SH-SY5Y) was exposed to toluene for the same period. Following exposure, cells were harvested and toxicity measured in terms of the following endpoints: membrane damage (LDH leakage), perturbations in intracellular free Ca 2+ , changes in glutathione redox status and dual-phosphorylation of MAP kinases ERK1/2, JNK and p38. The results show that sub-chronic exposure to the volatile organic solvents causes membrane damage, increased intracellular free calcium and altered glutathione redox status in both cell lines. However, acute and sub-chronic solvent exposure did not result in MAP kinase phosphorylation. Toxicity of the solvents tested increased with hydrophobicity. The lowest-observed-adverse-effect-levels (LOAELs) measured in vitro were close to blood solvent concentrations reported for individuals exposed to the agents at levels at or below their individual threshold limit values (TLVs)

IN VITRO PHYTOREMEDIATION OF PERSISTENT ORGANIC POLLUTANTS BY Helianthus annuus L. PLANTS

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Marcos V. de Almeida

Full Text Available Plant model systems are needed to properly conduct basic laboratory studies prior to field applications of phytoremediation. In vitro plant cultures are a useful tool for such research. This study focuses on the removal and/or degradation of 24 persistent organic pollutants under in vitro conditions by Helianthus annuus L (sunflower. The main purpose of exploiting this plant for phytoremediation process is due to its strong adaptability to adverse environments conditions such as resistance to pests, disease, and others. The study of bioremediation effects of all chemical molecules under in vitro conditions showed promising results. Sixteen out of twenty-four compounds evaluated reached up to 87% for remediation. The highest accumulation of pollutants was observed in the roots, showing that these results are consistent with the current literature. Through the study, it was observed effective absorption of POPs with logKow ranging from 4.50 to 6.91. Sunflower phytoremediation process efficiently detected heptachlor, aldrin, heptachlor epoxide, trans-chlordane, chlordane, dieldrin, DDE, DDT, methoxychlor, mirex and decachlorobiphenyl.

Nonconserved Residues Ala287 and Ser290 of the Cryptosporidium hominis Thymidylate Synthase Domain Facilitate Its Rapid Rate of Catalysis

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Doan,L.; Martucci, W.; Vargo, M.; Atreya, C.; Anderson, K.

2007-01-01

Cryptosporidium hominis TS-DHFR exhibits an unusually high rate of catalysis at the TS domain, at least 10-fold greater than those of other TS enzymes. Using site-directed mutagenesis, we have mutated residues Ala287 and Ser290 in the folate-binding helix to phenylalanine and glycine, respectively, the corresponding residues in human and most other TS enzymes. Our results show that the mutant A287F, the mutant S290G, and the double mutant all have reduced affinities for methylene tetrahydrofolate and reduced rates of reaction at the TS domain. Interestingly, the S290G mutant enzyme had the lowest TS activity, with a catalytic efficiency {approx}200-fold lower than that of the wild type (WT). The rate of conformational change of the S290G mutant is {approx}80 times slower than that of WT, resulting in a change in the rate-limiting step from hydride transfer to covalent ternary complex formation. We have determined the crystal structure of ligand-bound S290G mutant enzyme, which shows that the primary effect of the mutation is an increase in the distance between the TS ligands. The kinetic and crystal structure data presented here provide the first evidence explaining the unusually fast TS rate in C. hominis.

Effect of organically bound tritium (OBT) on pre-implantation mouse embryos in vitro

International Nuclear Information System (INIS)

Yamada, Takeshi; Ohyama, Harumi

1989-01-01

Effect of organically bound tritium (OBT), such as tritiated thymidine and tritium-labeled amino acids, on mouse preimplantation embryos was examined in vitro. Mouse zygotes fertilized in vitro (BC3F 1 eggs x ICR sperm) were cultured in the media containing OBT in various concentrations up to the blastocyst stage. The LD 50 in terms of tritium concentrations in the culture medium were determined by measuring tritium concentrations in the medium to inhibit 50 % of embryos to form blastocyst in vitro. Tritium activities in the embryos were measured at various times during culture of embryos at LD 50 concentration in order to estimate absorbed radiation dose in embryonic cells. The LD 50 values obtained indicate that OBT could inhibit the embryonic development 1000 times more effectively that tritiated water (HTO). However, differences in LD 50 values in terms of absorbed radiation dose between OBT and HTO is not so essential, and might be explained by localized spatial distribution of OBT within the cell. (author)

Frequency of Mycoplasma hominis and Ureaplasma urealyticum infections in women with systemic lupus erythematosus Freqüência da infecção pelo Mycoplasma hominis e Ureaplasma urealyticum em mulheres portadoras de lupus eritematoso sistêmico

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Alcyone A. Machado

2001-06-01

Full Text Available Ureaplasma urealyticum (UU and Mycoplasma hominis (MH have been detected in the urine of women with systemic lupus erythematosus (SLE. We evaluated the presence of these mycoplasma in the endocervix of women presenting SLE. A total of 40 SLE patients (mean age 40.2 years, and 51 healthy women (mean age 30.9 years, were studied. Endocervical swabs were cultured in specific liquid media for MH or UU, detected by a quantitative color assay, and considered positive at >10³ dilutions. Statistical analysis was performed using the two-tailed Fisher test. UU was detected in 52.5 % of patients and in 11.8% of controls (p= 0.000059. MH was detected in 20% of patients and 2% controls (p=0.003905. Both mycoplasmas were detected in 7.3% patients and 0% controls (pUreaplasma urealyticum (UU e Mycoplasma hominis (MH têm sido detectados em urina de mulheres com lupus eritematoso sistêmico (LES. Avaliamos a presença destes mycoplasmas no endocervix de mulheres apresentando LES. Um total de 40 pacientes com LES (idade média de 40,2 anos, e 51 mulheres sadias (idade média de 30.9 anos, foram estudadas. Swabs do endocervix foram cultivados em meio líquido específico para MH e UU, detectados por teste colorimétrico quantitativo, considerando positivo diluições > 10³ . Análise estatística foi feita usando teste de Fisher. UU foi detectado em 52,5% das pacientes e em 11,8% dos controles (p= 0.000059. MH foi detectado em 20% das pacientes e 2% dos controles (p=0.003905. Ambos mycoplasmas foram detectados em 7,3 % das pacientes e 0% dos controles (p<0.000001. Os resultados aqui reportados corroboram com a associação de infecção por mycoplasma e LES. Estes agentes podem estimular a produção de clones autoreativos.

Effects of Hominis Placenta Aqua-acupuncture on Kidney and Liver Intoxicated by HgCI2 in Rats

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Lee, Sang-Keel

2000-12-01

Full Text Available Objective: This study was perfomled to examine the therapeutic effect of aqua-acupuncture solution of Hominis Placenta(HP on kidney and liver intoxicated by HgCl2 in rats. Methods: 10% and 25% HP aqua-acupuncture were carried out everyday for 8 days on corresponding bilateral loci of Shinsu(BL23 and Kansu(BL18, respectively, after mercuric chloride intoxication in rats. Thereafter BUN, creatinine, GOT, GPT, ALP, -GT, albumin and total bilirubin were measured before intoxication, and at the 4th and the 8th experimental day. Histopathological and immunochemical observation were also carried out. Results: 1. It showed significant decreases of BUN in the group of 10% HP aqua-acupuncture into Shinsu on the 4th experimental day as compared with the control group. 2. It showed significant decreases of creatinine in the group of 10% HP aqua-acupuncture into Shinsu on the 4th and the 8th experimental days as compared with the control group. 3. There were not any significant changes of GOT, GPT, ALP, γ-GT, albumin and total bilirubin in the HP aqua-acupuncture groups compared with the control group. 4. By the histopathological observations on kidney under a light microscope, all the 10% and 25% HP aqua-acupuncture into Shinsu showed the preventive effect on tubulo-interstitial necrosis and muItifocal calcification in tubular lumen respectively compared with the control group. 5. By the histopathological observations on liver under a light mIcroscope, the groups 10% and 25% HP aqua-acupuncture into Kansu did not show any significant changes in the liver compared with the control group. 6. By the immunochemical analysis of heat shock protein(hsp and glucose-regulated protein(grp in rat renal cortex, the expressions of hsp70 and grp78 were decreased in the and HP aqua-acupuncture into Shinsu respectively compared with the control group. Conclusion: These results suggest that Hominis Placenta aqua-acupuncture have an effect on prevention and protection of

Comparison of the in vivo and in vitro genotoxicity of glyphosate isopropylamine salt in three different organisms.

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Alvarez-Moya, Carlos; Silva, Mónica Reynoso; Ramírez, Carlos Valdez; Gallardo, David Gómez; Sánchez, Rafael León; Aguirre, Alejandro Canales; Velasco, Alfredo Feria

2014-03-01

There is considerable controversy with regard to the genotoxicity of glyphosate, with some reports stating that this compound is non-toxic for fish, birds and mammals. In this work, we used the comet assay to examine the genotoxicity of glyphosate isopropylamine (0.7, 7, 70 and 700 μM) in human lymphocytes, erythrocytes of Oreochromis niloticus and staminal nuclei of Tradescantia (4430) in vitro and in vivo. Cells, nuclei and fish that had and had not been exposed to 5 mM N-nitrosodiethylamine (NDEA) were used as positive and negative controls, respectively. Significant (p cell types and organisms tested. Human lymphocytes and Tradescantia hairs showed lower genetic damage in vivo compared to in vitro, possibly because of efficient metabolization of the herbicide. In O. niloticus erythrocytes, significant (p cells and organisms studied at concentrations of 0.7-7 μM.

Correlation between organic acid exudation and metal uptake by ectomycorrhizal fungi grown on pond ash in vitro

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Ray, P.; Adholeya, A. [Energy & Resources Institute, New Delhi (India). India Habitat Centre

2009-04-15

Experiments were conducted to investigate the effect of coal ash on organic acid exudation and subsequent metal uptake by ectomycorrhizal fungi. Four isolates of ectomycorrhizal fungi namely, Pisolithus tinctorius (EM-1293 and EM-1299), Scleroderma verucosum (EM-1283) and Scleroderma cepa (EM-1233) were grown on pond ash moistened with Modified Melin-Norkans medium in vitro. Exudation of formic acid, malic acid and succinic acid by these fungi were detected by HPLC. Mycelial accumulation of Al, As, Cd, Cr, Ni and Pb by these fungi was assayed by atomic absorption spectrophotometer. Relationship between organic acid exudation and metal uptake was determined using classical multivariate linear regression model. Correlation between organic acid exudation and metal uptake could be substantiated when several metals are considered collectively. The finding supports the widespread role of low molecular weight organic acid as a function of tolerance, when exposed to metals in vitro.

DNA sequencing reveals limited heterogeneity in the 16S rRNA gene from the rrnB operon among five Mycoplasma hominis isolates

DEFF Research Database (Denmark)

Mygind, T; Birkelund, Svend; Christiansen, Gunna

1998-01-01

To investigate the intraspecies heterogeneity within the 16S rRNA gene of Mycoplasma hominis, five isolates with diverse antigenic profiles, variable/identical P120 hypervariable domains, and different 16S rRNA gene RFLP patterns were analysed. The 16S rRNA gene from the rrnB operon was amplified...... by PCR and the PCR products were sequenced. Three isolates had identical 16S rRNA sequences and two isolates had sequences that differed from the others by only one nucleotide....

Treatment for intractable anemia with the traditional Chinese medicines Hominis Placenta and Cervi Cornus Colla (deer antler glue

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Yasuyo Hijikata

2009-05-01

Full Text Available Yasuyo Hijikata1, Takashi Kano2, Lu Xi31Toyodo Hijikata Clinic, Osaka, Japan; 2Kano Clinic, Osaka city, Osaka, Japan; 3Traditional Chinese Medicine Institute, Si-chuan Province, ChinaObjective: Intractable anemia, such as aplastic anemia or that presumably associated with chronic herpes virus infections, sometimes require bone marrow transplant. We investigated the use of traditional Chinese medicine (TCM for the treatment of intractable anemia. Method: Placenta Hominis (PH, steam boiled and roasted, and Cervi Cornus Colla (deer antler glue has been used in China for hundreds of years to treat anemia. After consent was obtained, we prescribed these two materials for a 74-year-old female with aplastic anemia and a 26-year-old male with presumably a virus-induced anemia. Concomitant conventional therapy was continued in both patients as prescribed by their respective attending physicians. Conclusion: Conventional therapy with steroid hormones, immunosuppressive drugs, platelet and erythrocyte transfusions were not effective in these patients. In addition, both patients suffered from serious side effects. In two patients, ingestion of Placenta Hominis and Cervi Cornus Colla with TCM prescriptions increased the platelet and enhanced the hemoglobin concentration in several months of therapy accompanied by a dramatic improvement in quality of life. The addition to conventional therapy of PH and Cervi Cornus Colla, the latter of which is very easy to obtain, may be one of the potentially advantageous choices in case of otherwise intractable anemia.Keywords: placenta, antler glue, Cervi Cornus Colla, anemia, aplastic anemia

Comparative in vitro activities of investigational peptide deformylase inhibitor NVP LBM-415 and other agents against human mycoplasmas and ureaplasmas.

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Waites, Ken B; Reddy, Nipun B; Crabb, Donna M; Duffy, Lynn B

2005-06-01

Peptide deformylase inhibitor LBM-415 and seven other drugs were tested against Mycoplasma pneumoniae (100 isolates), Mycoplasma hominis (20 isolates), Mycoplasma fermentans (10 isolates), and Ureaplasma species (50 isolates). LBM-415 was active against M. pneumoniae (MICs, hominis and M. fermentans and modest activity against Ureaplasma spp.

In vitro to in vivo extrapolation of biotransformation rates for assessing bioaccumulation of hydrophobic organic chemicals in mammals.

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Lee, Yung-Shan; Lo, Justin C; Otton, S Victoria; Moore, Margo M; Kennedy, Chris J; Gobas, Frank A P C

2017-07-01

Incorporating biotransformation in bioaccumulation assessments of hydrophobic chemicals in both aquatic and terrestrial organisms in a simple, rapid, and cost-effective manner is urgently needed to improve bioaccumulation assessments of potentially bioaccumulative substances. One approach to estimate whole-animal biotransformation rate constants is to combine in vitro measurements of hepatic biotransformation kinetics with in vitro to in vivo extrapolation (IVIVE) and bioaccumulation modeling. An established IVIVE modeling approach exists for pharmaceuticals (referred to in the present study as IVIVE-Ph) and has recently been adapted for chemical bioaccumulation assessments in fish. The present study proposes and tests an alternative IVIVE-B technique to support bioaccumulation assessment of hydrophobic chemicals with a log octanol-water partition coefficient (K OW ) ≥ 4 in mammals. The IVIVE-B approach requires fewer physiological and physiochemical parameters than the IVIVE-Ph approach and does not involve interconversions between clearance and rate constants in the extrapolation. Using in vitro depletion rates, the results show that the IVIVE-B and IVIVE-Ph models yield similar estimates of rat whole-organism biotransformation rate constants for hypothetical chemicals with log K OW  ≥ 4. The IVIVE-B approach generated in vivo biotransformation rate constants and biomagnification factors (BMFs) for benzo[a]pyrene that are within the range of empirical observations. The proposed IVIVE-B technique may be a useful tool for assessing BMFs of hydrophobic organic chemicals in mammals. Environ Toxicol Chem 2017;36:1934-1946. © 2016 SETAC. © 2016 SETAC.

Microsporidia and Cryptosporidium in horses and donkeys in Algeria: detection of a novel Cryptosporidium hominis subtype family (Ik) in a horse.

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Laatamna, Abd Elkarim; Wagnerová, Pavla; Sak, Bohumil; Květoňová, Dana; Xiao, Lihua; Rost, Michael; McEvoy, John; Saadi, Ahmed Rachid; Aissi, Meriem; Kváč, Martin

2015-03-15

A total of 219 and 124 individual fecal samples of horses and donkeys, respectively, were screened for the presence of Cryptosporidium spp., Encephalitozoon spp., and Enterocytozoon bieneusi DNA by genus-specific nested PCR. Isolates were genotyped by sequence analysis of SSU rRNA, GP60, TRAP-C1, COWP, and HSP70 loci in Cryptosporidium, and the ITS region in microsporidia. Cryptosporidium spp. was detected on 3/18 horse farms and 1/15 farms where donkeys were kept. Overall, five (2.3%) horse and two (1.6%) donkey specimens were PCR positive for Cryptosporidium. Genotyping at SSU and GP60 loci revealed that three isolates from horses and donkeys were C. parvum subtype family IIaA16G1R1, one isolate from a horse was, C. muris RN66, and one isolate from a donkey was C. muris TS03. An isolate from a horse shared 99.4% and 99.3% similarity with Cryptosporidium hominis and C. cuniculus, respectively, at the SSU locus. This isolate shared 100% identity with C. hominis at the TRAP-C1, COWP, and HSP70 loci, and it was from the novel gp60 subtype family IkA15G1. Microsporidia were found on 6/18 horse and 2/15 donkey farms. E. bieneusi was identified in 6.8% (15/219) and 1.6% (2/124), and Encephalitozoon cuniculi was identified in 1.8% (4/219) and 1.6% (2/124), of horses and donkeys, respectively. Three genotypes of E. cuniculi (I, II and III) were detected in horses, and E. cuniculi genotype II was detected in donkeys. Four genotypes of E. bieneusi (horse1, horse 2, CZ3, D) were described in horses. An additional five horses and two donkeys were positive for E. bieneusi, but the isolated were not genotyped. Neither Cryptosporidium nor microsporidia prevalence were affected by sex, age, type of breeding, or whether the host was a horse or a donkey. Copyright © 2015 Elsevier B.V. All rights reserved.

3D bioprinting matrices with controlled pore structure and release function guide in vitro self-organization of sweat gland.

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Liu, Nanbo; Huang, Sha; Yao, Bin; Xie, Jiangfan; Wu, Xu; Fu, Xiaobing

2016-10-03

3D bioprinting matrices are novel platforms for tissue regeneration. Tissue self-organization is a critical process during regeneration that implies the features of organogenesis. However, it is not clear from the current evidences whether 3D printed construct plays a role in guiding tissue self-organization in vitro. Based on our previous study, we bioprinted a 3D matrix as the restrictive niche for direct sweat gland differentiation of epidermal progenitors by different pore structure (300-μm or 400-μm nozzle diameters printed) and reported a long-term gradual transition of differentiated cells into glandular morphogenesis occurs within the 3D construct in vitro. At the initial 14-day culture, an accelerated cell differentiation was achieved with inductive cues released along with gelatin reduction. After protein release completed, the 3D construct guide the self-organized formation of sweat gland tissues, which is similar to that of the natural developmental process. However, glandular morphogenesis was only observed in 300-μm-printed constructs. In the absence of 3D architectural support, glandular morphogenesis was not occurred. This striking finding made us to identify a previously unknown role of the 3D-printed structure in glandular tissue regeneration, and this self-organizing strategy can be applied to forming other tissues in vitro.

Large outbreak of Cryptosporidium hominis infection transmitted through the public water supply, Sweden.

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Widerström, Micael; Schönning, Caroline; Lilja, Mikael; Lebbad, Marianne; Ljung, Thomas; Allestam, Görel; Ferm, Martin; Björkholm, Britta; Hansen, Anette; Hiltula, Jari; Långmark, Jonas; Löfdahl, Margareta; Omberg, Maria; Reuterwall, Christina; Samuelsson, Eva; Widgren, Katarina; Wallensten, Anders; Lindh, Johan

2014-04-01

In November 2010, ≈27,000 (≈45%) inhabitants of Östersund, Sweden, were affected by a waterborne outbreak of cryptosporidiosis. The outbreak was characterized by a rapid onset and high attack rate, especially among young and middle-aged persons. Young age, number of infected family members, amount of water consumed daily, and gluten intolerance were identified as risk factors for acquiring cryptosporidiosis. Also, chronic intestinal disease and young age were significantly associated with prolonged diarrhea. Identification of Cryptosporidium hominis subtype IbA10G2 in human and environmental samples and consistently low numbers of oocysts in drinking water confirmed insufficient reduction of parasites by the municipal water treatment plant. The current outbreak shows that use of inadequate microbial barriers at water treatment plants can have serious consequences for public health. This risk can be minimized by optimizing control of raw water quality and employing multiple barriers that remove or inactivate all groups of pathogens.

[Study on the inhibitory activity, in vitro, of baicalein and baicalin against skin fungi and bacteria].

Science.gov (United States)

Yang, D; Hu, H; Huang, S; Chaumont, J P; Millet, J

2000-05-01

In this paper, we concentrated in examining, in vitro, the antiseptic activity of the baicalein and baicalin upon the seventeen pathogenic skin fungal and sixteen skin bacterial strains, these two flavonic compounds were known principally as the biosubstances of a traditional Chinese medicinal plant: Scutellaria baicalensis. In agar media, the baicalein possessed potent specific activity against the pathogenic yeasts with MICs of 70-100 micrograms/ml; But in the same condition, no inhibitory effect was observed upon dermatophytes and filamentous imperfect fungi for baicalein, and upon all used strains for baicalin. According to the antibacterial test of baicalein, a high efficacy was achieved against certain causative specie of axillary and foot's odour such as Micrococcus sedentarius, Staphylococcus epidermidis, S. hominis and C. xerosis with a MICs inferior to 250 micrograms/ml. The good inhibitory activity of baicalein could be linked to the group hydroxyl (-OH) in position seven of the molecule.

Clinical Review of the Effects of Hominis Placental Pharmacopuncture in the Treatment of Facial Spasm Patients

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Jo Na-Young

2013-09-01

Full Text Available Objectives: The main purpose of this research is to investigate the effect of treatment with Hominis Placental pharmacopuncture (HPP for 32 patients with hemifacial spasm. Methods: We treated facial spasm patients with acupuncture and HPP at Sabaek (ST2, Seung-eup (ST1, Gwallyeo (SI18, Chanjuk (BL2, Sajukgong (TE23, Hagwan (ST7, Hyeopgeo (ST6, Jichang (ST4, Wan-gol (SI4 and Yepung (TE17, and we investigated the effect by using Scott’s scale. The data were analyzed by using the SPSS/10.0 for windows program with descriptive statistics, the paired t-test, and the Shapiro-Wilk normality test. Results: After treatment, the grade of the spasm’s intensity based on Scott’s description were decreased significantly. About 72% of the patients felt that the combination treatment had produced excellent results. Conclusion: These data suggested that HPP can be useful for treating facial spasm patients.

Therapeutic effects of Hominis placenta herb-acupuncture in adjuvant-induced arthritis rat

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MiJung Yeom

2002-02-01

Full Text Available Rheumatoid arthritis (RA is a chronic inflammatory autoimmune disease, characterized by leukocyte infiltration, a chronic inflammation of the joint, a pannus formation and the extensive destruction of the articular cartilage and bone. Several proinflammatory cytokines such as tumor necrosis factor-α (TNF-α, interleukin-1β (IL-1β and interleukin 6 (IL-6 have been implicated in the pathological mechanisms of synovial tissue proliferation, joint destruction and programmed cell death in rheumatoid joint. In the Korean traditional medicine, Hominis placenta (HP as an herbal solution of herb-acupuncture has been widely used to treat the inflammatory diseases including RA. In order to study the medicinal effect of HP herb-acupuncture on rheumatoid joint, an adjuvant-induced arthritis (AIA was generated by the injection of 1.5 mg of Mycobacterium tuberculosis, emulsified in squalene, to the base of the tail of Spraque-Dawley (SD rats. After onset stage of polyarthritis, HP was daily injected to the Zusanli (ST36 acupuncture points in both of rat lags and the expression patterns of cytokines such as TNF-α, IL-1β, and IL-6 at the knee joint were analyzed using immunostaining and RT-PCR. The HP herb-acupuncture was found to be effective to alleviate the arthritic symptoms in adjuvant-induced arthritic rats as regards the joint appearance and the expression profiles of inflammatory cytokines. In conclusion, therapeutic effects of HP herb-acupuncture on the rat with AIA might be related to anti-inflammatory activities of the hurb-acupuncture.

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model.

Science.gov (United States)

Chang, Robert; Emami, Kamal; Wu, Honglu; Sun, Wei

2010-12-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

International Nuclear Information System (INIS)

Chang, Robert; Sun Wei; Emami, Kamal; Wu Honglu

2010-01-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

Energy Technology Data Exchange (ETDEWEB)

Chang, Robert; Sun Wei [Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, PA (United States); Emami, Kamal; Wu Honglu, E-mail: rcc34@drexel.ed, E-mail: sunwei@drexel.ed, E-mail: kamal.emami-1@nasa.go, E-mail: honglu.wu-1@nasa.go [Radiation Biophysics Laboratory, Human Adaptation and Countermeasures Office, NASA Johnson Space Center, Houston, TX (United States)

2010-12-15

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically

Inquiry of cases of myiasis by Dermatobia hominis in dogs of the southern zone of Rio de Janeiro municipality in 2000

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Bianca Chiganer Cramer-Ribeiro

2002-01-01

Full Text Available Na Zona Sul do Município do Rio de Janeiro, foi realizado um inquérito sobre os casos de miíases por larvas de Dermatobia hominis (berne em cães atendidos em 34 clínicas e consultórios veterinários durante 2000, para identificar fatores predisponentes a esta parasitose e contribuir na compreensão dos seus aspectos epidemiológicos e na sua prevenção. Pelo menos um caso foi atendido em 24 estabelecimentos, sendo os cães de raça definida, adultos, machos e de pelagem curta e clara os mais acometidos. Os locais do corpo mais afetados foram aqueles de fácil acesso às moscas: membros, dorso, região lombar, cabeça e pescoço. Na maioria dos casos, não foram observadas complicações durante e após o tratamento e o tempo de cura foi menor que cinco dias. Como não se observou determinada época do ano com maior ocorrência de casos, deve-se realizar programas preventivos durante o ano todo, baseados em dados epidemiológicos como as características fenotípicas (raça, cor e comprimento de pelagem, idade e sexo dos animais acometidos. Os proprietários devem ser encorajados a procurar por orientação veterinária quando houver larvas nos animais e a manter boas condições higiênicas ambientais. A prevenção deve ser aplicada principalmente nas áreas que propiciam um ambiente favorável à sobrevivência e proliferação da D. hominis e das moscas vetoras de seus ovos, pois a maior parte dos animais afetados adquiriu a parasitose em viagens a sítios fora da área estudada, possibilitando que casos de berne fossem observados em cães residentes em uma área predominantemente urbana como a Zona Sul.

Comparison of the in vivo and in vitro genotoxicity of glyphosate isopropylamine salt in three different organisms

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Carlos Alvarez-Moya

2014-01-01

Full Text Available There is considerable controversy with regard to the genotoxicity of glyphosate, with some reports stating that this compound is non-toxic for fish, birds and mammals. In this work, we used the comet assay to examine the genotoxicity of glyphosate isopropylamine (0.7, 7, 70 and 700 µM in human lymphocytes, erythrocytes of Oreochromis niloticus and staminal nuclei of Tradescantia (4430 in vitro and in vivo. Cells, nuclei and fish that had and had not been exposed to 5 mM N-nitrosodiethylamine (NDEA were used as positive and negative controls, respectively. Significant (p 7 µM, whereas in vitro, glyphosphate was genotoxic in human lymphocytes and Tradescantia hairs at > 0.7 µM. These results indicate that glyphosate is genotoxic in the cells and organisms studied at concentrations of 0.7-7 µM.

阴道毛滴虫与人型支原体共生对铁氧还蛋白基因影响%Effects of the Symbiosis of Trichomonas Vaginalis with Mycoplasma Hominis on Ferredoxin Gene

Institute of Scientific and Technical Information of China (English)

刘晓东; 温雯静; 薛长贵

2011-01-01

We isolated 30 Trichomonas vaginalis for the PCR detection from the gynecological outpatients in the Affiliated Hospital of Zhengzhou University using the specific 16s rDNA primers of Mycoplasma hominis. The results showed that there were 25 cases of Mycoplasma hominis infection, with the infection rate of 83. 33%. This gave a clew that the symbiosis of Trichomonas vaginalis with Mycoplasma hominis may be of certain generality in China. We sequenced the ferredoxin gene of 10 Trichomonas vaginalis where 5 Mycoplasma hominis were positive and five negative, and found that the ferredoxin (Fd) gene of the 10 Trichomonas vaginalis were exactly the same. But compared to the genes in the GenBank. A comparative analysis of the gene revealed that there were 3 more ctg bases at the 200th position of encoding leucine, but this did not lead to changes in reading frame. The gene homology was 99%.%采用人型支原体16s rDNA的特异引物,对从郑州大学附属医院妇科门诊患者分离到的30株阴道毛滴虫进行PCR检测,结果有25株感染人型支原体,感染率为83.33%,这显示了阴道毛滴虫和人型支原体之间的共生关系在中国具有普遍性.并对10株阴道毛滴虫(5株人型支原体阳性和5株人型支原体阴性)的铁氧还蛋白(Fd)基因进行测序,探讨人型支原体对Fd基因的影响,结果发现10株阴道毛滴虫的Fd基因完全相同,但与GenBank中的基因进行比较分析,在第200位多出ctg三个碱基,编码亮氨酸,但并未导致读码框架改变.基因同源性为99%.

Endocrine activity of persistent organic pollutants accumulated in human silicone implants — Dosing in vitro assays by partitioning from silicone

DEFF Research Database (Denmark)

Gilbert, Dorothea; Mayer, Philipp; Pedersen, Mikael

2015-01-01

Persistent organic pollutants (POPs) accumulated in human tissues may pose a risk for human health by interfering with the endocrine system. This study establishes a new link between actual human internal POP levels and the endocrine active dose in vitro, applying partitioning-controlled dosing...

Effects of compost organic amendments on chemical composition and in vitro digestibility of alfalfa (Medicago sativa L.

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Francesco Montemurro

2010-01-01

Full Text Available The following fertiliser treatments were compared during the years 2002 and 2003 on alfalfa forage (Medicago sativa L.: compost obtained from the organic fraction of the Municipal Solid Waste (MSW; olive pomace compost (OPC; mineral fertiliser (Min. All the treatments allowed a distribution of 75kg ha-1 of P2O5. Three cuttings occurred: at 168, 206 and 351 days after compost application (DAA in 2002; 119, 152 and 320 DAA in 2003. Cumulative biomass and dry matter yields were measured during each experimental year. Furthermore, chemical composition and in vitro digestibility of dry matter (DMd, organic matter (OMd, crude protein (CPd and NDF (NDFd were determined. MSW treatment showed a significantly (P<0.01 higher content of ADL than OPC and Min (77.0, 66.0 and 65.0g kg-1 DM, respectively. Fertiliser treatments also affected (P<0.01 digestibility parameters. In fact, DMd and OMd values showed the same trend with lower percentages in MSW treatment than in the OPC and Min ones. The NDFd differed in all treatments having the highest value in OPC (40.1%. The results indicated that the soil distribution of organic materials offer the possibility to reduce the application of mineral fertilisers and production costs without decreasing alfalfa yield, forage chemical composition and in vitro digestibility.

Desenvolvimento das gônadas de Dermatobia hominis (Diptera: Cuterebridae

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Edy de Lello

1985-06-01

Full Text Available O trabalho descreve o desenvolvimento das gônadas do berne (D. hominis durante o período pupal. As pupas desenvolvidas de larvas com peso superior a 650 mg, deram imagos fêmeas, enquanto que as desenvolvidas daquelas pesando entre 500 e 650 mg deram macho, tendo havido um erro ao redor de 5%. Até o oitavo dia de pupação os testículos crescem mais que os ovários; a partir daí diminui o desenvolvimento, parando de crescer entre o vigésimo e vigésimo quinto dias. A espermatogênese inicia por volta do sétimo dia de pupa quando é grande o número de espermatócitos. No décimo dia alguns testículos apresentam considerável número de espermátides e os espermatozóides começam a aparecer por volta do vigésimo dia. A espermiogênese desenvolve-se sem interrupção e ao final da pupação quase toda loja testicular está repleta de espermatózóides. Os machos começam a nascer dois dias antes das fêmeas. Nessas, os ovaríolos aparecem formados por volta do oitavo dia de pupa; os folículos se individualizam por volta do vigésimo dia de pupa onde se distingue os trofócitos com núcleos politênicos e citoplasmas bem basófilos, enquanto o ovócito tem citoplasma mais acidófilo e núcleo com cromatina bastante frouxa. A vitelogênese tem início ao redor do vigésimo quinto dia de pupa e se completa ao nascimento da imago. A ligação das gônadas com suas respectivas estruturas somáticas acontece ao redor do décimo terceiro dia de pupação.Gonadal development of Dermatobia hominis during pupal period is described. Pupae developing from larvae weighing above 650 mg turned into female flies whereas the ones from larvae weighing between 500 and 600 mg turned into males (with plus of minus 5% of error. Up to the 8th day of pupation the testes are larger than the ovaries; from that time on they decrease their speed of development and stop growing between the 20th and 25th day. The spermatogenesis starts around the 7th day when there

In vitro mouse spermatogenesis with an organ culture method in chemically defined medium.

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Hiroyuki Sanjo

Full Text Available We previously reported the successful induction and completion of mouse spermatogenesis by culturing neonatal testis tissues. The culture medium consisted of α-minimum essential medium (α-MEM, supplemented with Knockout serum replacement (KSR or AlbuMAX, neither of which were defined chemically. In this study, we formulated a chemically defined medium (CDM that can induce mouse spermatogenesis under organ culture conditions. It was found that bovine serum albumin (BSA purified through three different procedures had different effects on spermatogenesis. We also confirmed that retinoic acid (RA played crucial roles in the onset of spermatogonial differentiation and meiotic initiation. The added lipids exhibited weak promoting effects on spermatogenesis. Lastly, luteinizing hormone (LH, follicle stimulating hormone (FSH, triiodothyronine (T3, and testosterone (T combined together promoted spermatogenesis until round spermatid production. The CDM, however, was not able to produce elongated spermatids. It was also unable to induce spermatogenesis from the very early neonatal period, before 2 days postpartum, leaving certain factors necessary for spermatogenic induction in mice unidentified. Nonetheless, the present study provided important basic information on testis organ culture and spermatogenesis in vitro.

In vitro metabolism and permeation studies in rat jejunum

DEFF Research Database (Denmark)

Gammelgaard, Bente; Jensen, K; Steffansen, B

1999-01-01

The purpose of these studies was to compare the in vitro absorption of two inorganic chromium(III) compounds: chromium chloride and chromium nitrate, with organic chromium(III)-picolinate; and to investigate if any in vitro metabolism of chromium(VI) takes place. The in vitro metabolism studies...

Comparing the effect of homogenization and heat processing on the properties and in vitro digestion of milk from organic and conventional dairy herds.

Science.gov (United States)

Van Hekken, D L; Tunick, M H; Ren, D X; Tomasula, P M

2017-08-01

We compared the effects of homogenization and heat processing on the chemical and in vitro digestion traits of milk from organic and conventional herds. Raw milk from organic (>50% of dry matter intake from pasture) and conventional (no access to pasture) farms were adjusted to commercial whole and nonfat milk fat standards, and processed with or without homogenization, and with high-temperature-short-time or UHT pasteurization. The milk then underwent in vitro gastrointestinal digestion. Comparison of milk from organic and conventional herds showed that the milks responded to processing in similar ways. General composition was the same among the whole milk samples and among the nonfat milk samples. Protein profiles were similar, with intact caseins and whey proteins predominant and only minor amounts of peptides. Whole milk samples from grazing cows contained higher levels of α-linolenic (C18:3), vaccenic (C18:1 trans), and conjugated linoleic acids, and lower levels of palmitic (C16:0) and stearic (C18:0) acids than samples from nongrazing cows. Processing had no effect on conjugated linoleic acid and linolenic acid levels in milk, although homogenization resulted in higher levels of C8 to C14 saturated fatty acids. Of the 9 volatile compounds evaluated, milk from grazing cows contained lower levels of 2-butanone than milk from nongrazing cows, and milk from both farms showed spikes for heptanal in UHT samples and spikes for butanoic, octanoic, nonanoic, and N-decanoic acids in homogenized samples. At the start of in vitro digestion, nonfat raw and pasteurized milk samples formed the largest acid clots, and organic milk clots were larger than conventional milk clots; UHT whole milk formed the smallest clots. Milk digests from grazing cows had lower levels of free fatty acids than digests from nongrazing cows. In vitro proteolysis was similar in milk from both farms and resulted in 85 to 95% digestibility. Overall, milk from organic/grass-fed and conventional

Prevalence of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia lamblia among Young Children with and without Diarrhea in Dar es Salaam, Tanzania.

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Marit G Tellevik

Full Text Available Although enteroparasites are common causes of diarrheal illness, few studies have been performed among children in Tanzania. This study aimed to investigate the prevalence of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia lamblia among young children in Dar es Salaam, Tanzania, and identify risk factors for infection.We performed an unmatched case-control study among children 12 months (P = 0.003; OR = 3.5; 95% CI: 1.5-7.8. Among children aged 7-12 months, those who were breastfed had lower prevalence of G. lamblia infection than those who had been weaned (P = 0.012.Cryptosporidium infection is common among young Tanzanian children with diarrhea, particularly those living with HIV, and infection is more frequent during the rainy season. G. lamblia is frequently implicated in asymptomatic infections, but rarely causes overt diarrheal illness, and its prevalence increases with age.

Investigation of volatile organic biomarkers derived from Plasmodium falciparum in vitro

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Wong Rina PM

2012-09-01

Full Text Available Abstract Background There remains a need for techniques that improve the sensitive detection of viable Plasmodium falciparum as part of diagnosis and therapeutic monitoring in clinical studies and usual-care management of malaria infections. A non-invasive breath test based on P. falciparum-associated specific volatile organic compounds (VOCs could fill this gap and provide insights into parasite metabolism and pathogenicity. The aim of this study was to determine whether VOCs are present in the headspace above in vitro P. falciparum cultures. Methods A novel, custom-designed apparatus was developed to enable efficient headspace sampling of infected and non-infected cultures. Conditions were optimized to support cultures of high parasitaemia (>20% to improve the potential detection of parasite-specific VOCs. A number of techniques for VOC analysis were investigated including solid phase micro-extraction using two different polarity fibres, and purge and trap/thermal desorption, each coupled to gas chromatography–mass spectrometry. Each experiment and analysis method was performed at least on two occasions. VOCs were identified by comparing their mass spectra against commercial mass spectral libraries. Results No unique malarial-specific VOCs could be detected relative to those in the control red blood cell cultures. This could reflect sequestration of VOCs into cell membranes and/or culture media but solvent extractions of supernatants and cell lysates using hexane, dichloromethane and ethyl acetate also showed no obvious difference compared to control non-parasitized cultures. Conclusions Future in vivo studies analysing the breath of patients with severe malaria who are harbouring a parasite biomass that is significantly greater than achievable in vitro may yet reveal specific clinically-useful volatile chemical biomarkers.

Comparison of three in vitro methods for determining and predicting the organic matter digestibility of complete diets for ruminants

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Leeuw Klaas-Jan

2009-01-01

Full Text Available In this study, the organic matter digestibility (OMD of six complete diets for ruminants has been determined in-vivo in trials with sheep and in-vitro using two-stage Tilley and Terry (T&T method, gas production (GP technique and multi-enzyme incubation (EDOM procedures. The mean OMD values obtained in vivo and using T&T, GP and EDOM techniques were 684, 716, 685 and 710 g OM/kgDM respectively and did not differ significantly (P>0.05. The obtained in vitro results were regressed against determined in-vivo values to derive prediction equations. Using the T&T technique, the prediction equation OMD (in_vivo = -17.36 + 0.98 x OMD (in_vitro_T&T, (R2 = 0.75; RMSE = 37.59 has been obtained. The equation OMD (in_vivo = 198.98 + 0.71 x OMD (in_vitro_GP, (R2 = 0.21; RMSE = 66.36 has been derived for Gas production procedure, while the equation OMD (in_vivo = 102 + 0.82 x OMD (in_vitro_EDOM, (R2 = 0.86; RMSE = 27.30 has been generated for multi-enzyme incubation technique. The results of this study showed that the OMD of complete diets for ruminants can be successfully determined, and in-vivo values predicted, using multi-enzyme incubation procedure, which is important because of the fact that rumen liquor, needed for the in-vitro twostage T&T and GP techniques is not always available to analytical laboratories.

Desenvolvimento pós-embrionário do intestino anterior de Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae Post-embryonic development of Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae foregut

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Ana Maria Vieira

2001-03-01

Full Text Available Foregut in D. hominis (Linnaeus Jr., 1781 as the majority of the larval Diptera somatic tissue, is made up of polytenic cells, and grows at the expenses of the polytenization of its nuclei followed by the increase in size of each cell. The oesophagus, of ectodermic origem, is interiorly covered by a chitinous squamous epithelium that rests upon a very thin basal lamina. This sheet is surrounded by thick muscle bundles. The oesophagus intussuscepts the midgut forming the cardia. The cardia, with three epithelial layers: two internal ones, of ectodermal origin and one external of endodermic origin. At the anterior portion of the cardia, between these two types of epithelium, there is a cluster of small, non polytenic cells, forming the imaginal disk of the foregut. Metamoiphosis begins at the end of the larval period with signs of nuclear degeneration of all the polytenic cells, as well as the increase in number of the imaginal disk ones. The oesophagic portion intussuscepted into the cardia, everts; its cells suffer apoptosis and are replaced by the new cells growing from the imaginal disk. The external layer cells also degenerate and are pinched off into the lumen of the very anterior portion of the midgut. The newly formed oesophagus intussuscepts "de novo" to form the two internal layers of the adult cardia. At the same time the midgut regenerative cells grow anteriorly to form the new external layer of the adult cardia.

Comparing the impact of homogenization and heat processing on the properties and in vitro digestion of milk from organic and conventional dairy herds

Science.gov (United States)

The effects of homogenization and heat processing on the chemical and in vitro digestion traits of milk from organic and conventional herds were compared. Raw milk from organic (>50% of dry matter intake from pasture) and conventional (no access to pasture) farms were adjusted to commercial whole a...

Prevalence of Chlamydia trachomatis, Ureaplasma spp., Mycoplasma genitalium and Mycoplasma hominis among outpatients in central Greece: absence of tetracycline resistance gene tet(M over a 4-year period study

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A. Ikonomidis

2016-01-01

Full Text Available A total of 301 men and women attending local urologists and gynaecologists in the state of Thessaly, central Greece, were tested for Chlamydia trachomatis, Ureaplasma spp., Mycoplasma genitalium and Mycoplasma hominis DNA. Investigation of the tet(M gene, which confers tetracycline resistance in these genera, was also performed. Low incidence of C. trachomatis and Mycoplasma spp. as well as high prevalence of Ureaplasma spp., especially among women, were found. The tet(M gene was absent in all cases, notably in a region where doxycycline administration remains the first therapeutic option unless special medical conditions direct otherwise.

Gain in cellular organization of inflammatory breast cancer: A 3D in vitro model that mimics the in vivo metastasis

International Nuclear Information System (INIS)

Morales, Jorge; Alpaugh, Mary L

2009-01-01

The initial step of metastasis in carcinomas, often referred to as the epithelial-mesenchymal transition (EMT), occurs via the loss of adherens junctions (e.g. cadherins) by the tumor embolus. This leads to a subsequent loss of cell polarity and cellular differentiation and organization, enabling cells of the embolus to become motile and invasive. However highly malignant inflammatory breast cancer (IBC) over-expresses E-cadherin. The human xenograft model of IBC (MARY-X), like IBC, displays the signature phenotype of an exaggerated degree of lymphovascular invasion (LVI) in situ by tumor emboli. An intact E-cadherin/α, β-catenin axis mediates the tight, compact clump of cells found both in vitro and in vivo as spheroids and tumor emboli, respectively. Using electron microscopy and focused ion beam milling to acquire in situ sections, we performed ultrastructural analysis of both an IBC and non-IBC, E-cadherin positive cell line to determine if retention of this adhesion molecule contributed to cellular organization. Here we report through ultrastructural analysis that IBC exhibits a high degree of cellular organization with polar elements such as apical/lateral positioning of E-cadherin, apical surface microvilli, and tortuous lumen-like (canalis) structures. In contrast, agarose-induced spheroids of MCF-7, a weakly invasive E-cadherin positive breast carcinoma cell line, do not exhibit ultrastructural polar features. This study has determined that the highly metastatic IBC with an exaggerated malignant phenotype challenges conventional wisdom in that instead of displaying a loss of cellular organization, IBC acquires a highly structured architecture. These findings suggest that the metastatic efficiency might be linked to the formation and maintenance of these architectural features. The comparative architectural features of both the spheroid and embolus of MARY-X provide an in vitro model with tractable in vivo applications

Desenvolvimento pós-embrionário do intestino médio de Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae Post-embryonic development of Dermatobia hominis (Linnaeus Jr. (Diptera, Cuterebridae midgut

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Edy de Lello

2001-03-01

Full Text Available Dermatobia hominis (Linnaeus, 1781 midgut is internally lined by an epithelium of polytenic cells, some low others prismatic with well developed brush border. Their apical portion are enlarged by secretory vesicles, forming button-like structures that are pinched off to the lumen, some accompained by the nucleus characterizing apocrine and holocrine secretions. This epithelium is gradually renewed by small, non polytenic regenerative cells, found scattered at its basal portion. At the end of the third instar the metamorphosis begins. The epithelial cells present signs of degeneration and at the first day of pupation the regenerative cells increase in number. By the 5th day of pupation these regenerative cells, besides being increased in number, differentiate themselves into two layers: one similar to the dense conective tissue that sustainning the larval epithelium is pinched off to the midgut lumen forming the "yellow bodies"; the other, develops right under it as the imaginal epitelium. The disorganized muscles bundles of the midgut wall, are invaded by phagocytes. At the end of pupation the midgut has a low prismatic epithelium with brush-border. In the adult, the torax portion of the midgut has prismatic homogeneously basophilic epithelium while in the abdominal portion the epithelium is made of high prismatic cells full of small vacuoles. The larval midgut epithelium suffers programmed cell death non compatible with apoptose. During the metamorphosis the midgut lenght diminishes from 31mm in the larva to 14mm in the adult.

The effect of gamma irradiation on the digestibility of organic matter of poultry excreta (In vitro)

International Nuclear Information System (INIS)

Al-Masri, M.R.

1993-07-01

The changes in the digestibility of dry matter and organic matter by enzyme (in vitro) for two types of the excreta of laying hens were studied. In type I, excreta were dried at 170-180 C for 10 minutes whereas in type II dried at 55-60 C for several days. Each type was divided into two parts, the first stored for 3 months with the control. The second part was irradiated by gamma irradiation at 100 KGy and stored for 3 months with the control. The results indicated that there was significant (0.05) difference in the digestibility of dry matter and organic matter and the percentage of crude fibre between samples and the control for the types I and II before and after storage. The dry matter digestibility for types I and II increased by 7%, and the organic matter digestibility increased by 17% for type I and by 11% for type II before and after storage. The increase in the digestibility of dry matter and organic matter is attributed to the decrease in crude fibre obtained by irradiation. The storage of excreta after drying has no effects on the rate of increase in the digestibility of dry matter and organic matter due to irradiation in both types (I and II). (author). 19 refs., 5 figs., 4 tabs

Transfer of hydrophobic contaminants in urban runoff particles to benthic organisms estimated by an in vitro bioaccessibility test

DEFF Research Database (Denmark)

Nakajima, F.; Saito, K.; Isozaki, Y.

2006-01-01

An in vitro bioaccessibility test was applied for assessing the transfer of polycyclic aromatic hydrocarbons (PAHs) present in road dust, into benthic organisms living in a receiving water body. The road dust is supposed to be urban runoff particles under wet weather conditions. Sodium dodecyl...... sulfate (SDS) solution was used as a hypothetical gut fluid. Pyrene, fluoranthene and phenanthrene were the main PAH species in the SIDS extractable fraction of road dust, as well as the whole extract. Benzo(ghi)perylene showed relatively low concentrations in the SIDS extract in spite of a high...... concentration in the original dust. The PAH composition in benthic organisms (polychaetes) did not correspond with that of the surrounding sediment and the PAHs detected were also detected in high concentrations in the SDS extract of road dust. When testing the toxicity of the extracted contaminants...

In vitro proliferation of haemopoietic cells in the presence of adherent cell layers. II. Differential effect of adherent cell layers derived from various organs

NARCIS (Netherlands)

Reimann, J.; Burger, H.

1979-01-01

Mouse bone marrow-derived adherent cell populations promoted proliferation of haemopoietic cells in vitro in a liquid culture system for at least 4 weeks. Adherent cell layers derived from other haemopoietic organs (foetal liver, adult spleen) and fibroblasts from embryonic tissues did not maintain

In vitro systems in pneumocystis research

DEFF Research Database (Denmark)

Dei-Cas, E; Cailliez, J C; Lundgren, Bettina

1996-01-01

as well as elimination of host cell debris from parasite extracts. In other investigations, such as transmission, infectivity, life cycle, biochemical, in vitro culture or drug-screening studies, viable and infectious Pneumocystis organisms are urgently required. However, there is no generally accepted...

First Report of a Case of Pneumococcal Meningitis Which Did Not Respond to the Ceftriaxone Therapy despite the Isolated Organism Being Sensitive to This Antibiotic In Vitro

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Maryam Mojtabavi

2011-01-01

Full Text Available A 60-year-old man presented with pneumococcal meningitis which did not respond to the ceftriaxone therapy, in spite of in-vitro susceptibility (minimal inhibitory concentration of 0.016 μg/dLit of the isolated organism to this antibacterial agent, although ceftriaxone is still the drug of choice for such pneumococcal meningitis. Review of published articles revealed no report of clinical resistance in organisms which were susceptible to the same antimicrobial agent in vitro. This alarming emergence of isolates with in vivo resistance should be considered and even could lead to a shift in the empirical antibiotic therapy for pneumococcal infections.

Berberine Enhances the Antibacterial Activity of Selected Antibiotics against Coagulase-Negative Staphylococcus Strains in Vitro

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Robert D. Wojtyczka

2014-05-01

Full Text Available Synergistic interactions between commonly used antibiotics and natural bioactive compounds may exhibit therapeutic benefits in a clinical setting. Berberine, an isoquinoline-type alkaloid isolated from many kinds of medicinal plants, has proven efficacy against a broad spectrum of microorganisms. The aim of the presented work was to assess the antibacterial activity of berberine chloride in light of the effect exerted by common antibiotics on fourteen reference strains of Staphylococccus spp., and to evaluate the magnitude of interactions of berberine with these antistaphylococcal antibiotics. In our study minimum inhibitory concentrations (MIC of berberine chloride against CoNS ranged from 16 to 512 µg/mL. The most noticeable effects were observed for S. haemolyticus ATCC 29970, S. epidermidis ATCC 12228, S. capitis subsp. capitis ATCC 35661, S. galinarium ATCC 700401, S. hominis subsp. hominis ATCC 27844, S. intermedius ATCC 29663 and S. lugdunensis ATCC 49576. The most significant synergistic effect was noticed for berberine in combination with linezolid, cefoxitin and erythromycin. The synergy between berberine and antibiotics demonstrates the potential application of compound combinations as an efficient, novel therapeutic tool for antibiotic-resistant bacterial infections.

Berberine enhances the antibacterial activity of selected antibiotics against coagulase-negative Staphylococcus strains in vitro.

Science.gov (United States)

Wojtyczka, Robert D; Dziedzic, Arkadiusz; Kępa, Małgorzata; Kubina, Robert; Kabała-Dzik, Agata; Mularz, Tomasz; Idzik, Danuta

2014-05-22

Synergistic interactions between commonly used antibiotics and natural bioactive compounds may exhibit therapeutic benefits in a clinical setting. Berberine, an isoquinoline-type alkaloid isolated from many kinds of medicinal plants, has proven efficacy against a broad spectrum of microorganisms. The aim of the presented work was to assess the antibacterial activity of berberine chloride in light of the effect exerted by common antibiotics on fourteen reference strains of Staphylococccus spp., and to evaluate the magnitude of interactions of berberine with these antistaphylococcal antibiotics. In our study minimum inhibitory concentrations (MIC) of berberine chloride against CoNS ranged from 16 to 512 µg/mL. The most noticeable effects were observed for S. haemolyticus ATCC 29970, S. epidermidis ATCC 12228, S. capitis subsp. capitis ATCC 35661, S. galinarium ATCC 700401, S. hominis subsp. hominis ATCC 27844, S. intermedius ATCC 29663 and S. lugdunensis ATCC 49576. The most significant synergistic effect was noticed for berberine in combination with linezolid, cefoxitin and erythromycin. The synergy between berberine and antibiotics demonstrates the potential application of compound combinations as an efficient, novel therapeutic tool for antibiotic-resistant bacterial infections.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae. I. Development during the 3rd larval instar

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E. A. Gregorio

1990-09-01

Full Text Available The ultrastructure and distribution of gonial and somatic cells in the ovary of Dermatobia hominis was studied during the 3rd larval instar. In larvae weighing between 400 and 500 mg, the ovary is partially divided into basal and apical regions by oblong somatic cells that penetrate from the periphery; these cells show ovoid nucleus and cytoplasm full of microtubules. In both regions, gonial cells with regular outlines, large nucleus and low electron-density cytoplasm are scattered among the interstitial somatic cells. These later cells have small nucleus and electrodense cytoplasm. Clear somatic cells with small nucleus and cytoplasm of very low electron-density are restrict to the apical region of the gonad. Degenerating interstitial somatic cells are seen in the basal portion close to the ovary peduncle. During all this larval period the morphological features of the ovary remain almost the same. At the end of the period there is a gradual deposition of glycogen in the cytoplasm of the somatic cells, increase in the number and density of their mitochondria plus nuclear modification as membrane wrinkling and chromatin condensation in masses.

The Solubility of Cr-Organic Produced by Hydrolysis, Bioprocess and Bioremediation and its Effect on Fermented Rate, Digestibility and Rumen Microbe Population (in vitro

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UH Tanuwiria

2010-09-01

Full Text Available The research was conducted to study the production of organic chromium from the leather tanning waste and its effect on in vitro rumen fermentation activities. The research was divided into two phases. The first phase was production of organic chromium by alkali hydrolysis, S cereviceae bioprocess, and duckweed bioremediation that perceived solubility in neutral and acid solution. The second phase was the supplementation of organic-Cr in ration seen from in-vitro fermented rate, digestibility and microbe rumen population. Research was conducted experimentally using 4x4 factorial patterns, on the basis of Completely Randomized Design (CRD with three replications in each experimental unit. The first factor was the type of organic-Cr and the second factor was the supplement in ration at four levels, 1, 2, 3 and 4 ppm. The results of this research indicated that organic chromium can be synthesized by alkali hydrolysis, S cereviseae bioprocess and the activity of duckweed bioremediation. Among the three of processes referred, the highest level of Cr was obtained from S cereviseae bioprocess that was originated from leather-tanning waste. The levels of organic-Cr that was resulted from alkali hydrolysis, bioprocess from Cl3Cr.6H2O, bioprocess from Cr leather-tanning waste, and from duckweed bioremediation were 354, 1011, 3833 and 310 mg/kg, respectively. Organic-Cr characteristic of each product has relatively similar in ferment ability, dry matter and organic matter digestibility and rumen ecosystem. There is an indication that dry matter and organic matter digestibility and rumen microbe population in ration that was added with organic Cr from alkali hydrolysis was higher than other supplements. (Animal Production 12(3: 175-183 (2010Key Words: organic-Cr, rumen fermentation activities, rumen microbe population

Digestion of crude protein and organic matter of leaves by rumen microbes in vitro

Energy Technology Data Exchange (ETDEWEB)

Ciszuk, A.; Murphy, M.

1982-01-01

22 leaf specimens, of which 6 were from an energy-woods project, were studied by incubation in vitro with rumen microbes or pepsin-hydrochloric acid. Several were also examined in situ using the nylon-bag technique. Many leaves, despite their low fiber and high crude protein content, gave low values for organic matter digestibility. The crude protein degradation by rumen microbes or pepsin-hydrochloric acid was low, on average, compared with hay. There was a wide variation among leaf specimens. Variation was also found as regards ammonia production in short-term (4 hours) incubation. No close correlation was found between crude protein content and crude protein degradation, or between the estimates of ruminal degradation and of pepsin-hydrochloric acid digestibility. This suggest that there are leaves that gives ruminants substantial amounts of digestible protein yet escape ruminal fermentation. (Refs. 12).

Parasitoses intestinais de indígenas da comunidade Mapuera (Oriximiná, Estado do Pará, Brasil: elevada prevalência de Blastocystis hominis e encontro de Cryptosporidium sp e Cyclospora cayetanensis Intestinal parasitosis in Indians of the Mapuera community (Oriximiná, State of Pará, Brazil: high prevalence of Blastocystis hominis and finding of Cryptosporidium sp and Cyclospora cayetanensis

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Jaila Dias Borges

2009-06-01

Full Text Available Foi avaliada a ocorrência de parasitoses intestinais em indígenas da aldeia Mapuera (Oriximiná, Estado do Pará, Brasil. No contexto de apreciações congêneres, expressa contribuição para adequado conhecimento do assunto, significativo sob o ponto de vista médico-sanitário. O exame parasitológico das fezes, de 83 pessoas, realizado por meio de quatro métodos, pode ser considerado como dotado de razoável amplitude para estabelecer diagnósticos. Ocorreu encontro de cistos de protozoários e de ovos de helmintos de múltiplos tipos, até mesmo em expressivas porcentagens, merecendo destaque a muito freqüente presença de Blastocystis hominis (57,8%, como também o encontro de Cryptosporidium sp (3,6% e de Cyclospora cayetanensis (10,8%, comentado especificamente. O verificado demonstra que tais índios vivem em ambiente onde prevalecem más condições higiênicas, em especial, facilitador da disseminação de protozoários e helmintos pelo contato com o solo ou ingestão de água e alimentos contaminados.Occurrences of intestinal parasitosis in Indians of the Mapuera community (Oriximiná, State of Pará, Brazil were evaluated. Within the context of group assessment, this study makes a contribution towards adequate knowledge of this subject, which is significant from a medical-sanitary point of view. Parasitological examination of feces from 83 individuals, performed using four different methods, could be considered to have reasonable amplitude for establishing diagnoses. Protozoan cysts and helminth eggs of many types were found, even with significant percentages. The frequent presence of Blastocystis hominis (57.8%, along with findings of Cryptosporidium sp (3.6% and Cyclospora cayetanensis (10.8%, deserved highlighting with specific comments. The findings show that these Indians live in an environment in which poor hygiene conditions prevail. In particular, these facilitate the dissemination of protozoa and helminths through

Nuclear organization during in vitro differentiation of porcine mesenchymal stem cells (MSCs) into adipocytes.

Science.gov (United States)

Stachecka, Joanna; Walczak, Agnieszka; Kociucka, Beata; Ruszczycki, Błażej; Wilczyński, Grzegorz; Szczerbal, Izabela

2018-02-01

Differentiation of progenitor cells into adipocytes is accompanied by remarkable changes in cell morphology, cytoskeletal organization, and gene expression profile. Mature adipocytes are filled with a large lipid droplet and the nucleus tends to move to the cell periphery. It was hypothesized that the differentiation process is also associated with changes of nuclear organization. The aim of this study was to determine the number and distribution of selected components of nuclear architecture during porcine in vitro adipogenesis. The pig is an important animal model sharing many similarities to humans at the anatomical, physiological, and genetic levels and has been recognized as a good model for human obesity. Thus, understanding how cellular structures important for fundamental nuclear processes may be altered during adipocyte differentiation is of great importance. Mesenchymal stem cells (MSCs) were derived from bone marrow (BM-MSCs) and adipose tissue (AD-MSCs) and were cultured for 7 days in the adipogenic medium. A variable differentiation potential of these cell populations towards adipogenic lineage was observed, and for further study, a comparative characteristic of the nuclear organization in BM-MSCs and AD-MSCs was performed. Nuclear substructures were visualized by indirect immunofluorescence (nucleoli, nuclear speckles, PML bodies, lamins, and HP1α) or fluorescence in situ hybridization (telomeres) on fixed cells at 0, 3, 5, and 7 days of differentiation. Comprehensive characterization of these structures, in terms of their number, size, dynamics, and arrangement in three-dimensional space of the nucleus, was performed. It was found that during differentiation of porcine MSCs into adipocytes, changes of nuclear organization occurred and concerned: (1) the nuclear size and shape; (2) reduced lamin A/C expression; and (3) reorganization of chromocenters. Other elements of nuclear architecture such as nucleoli, SC-35 nuclear speckles, and telomeres

Synthetic biology projects in vitro.

Science.gov (United States)

Forster, Anthony C; Church, George M

2007-01-01

Advances in the in vitro synthesis and evolution of DNA, RNA, and polypeptides are accelerating the construction of biopolymers, pathways, and organisms with novel functions. Known functions are being integrated and debugged with the aim of synthesizing life-like systems. The goals are knowledge, tools, smart materials, and therapies.

Hemilucilia segmentaria (Fabricius, 1805 (Diptera: Calliphoridae as new biological vector of eggs of Dermatobia hominis (Linnaeus Jr., 1781 (Diptera: Oestridae in Reserva Biológica do Tinguá, Rio de Janeiro, Brazil

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Marinho Clarissa Rezende

2003-01-01

Full Text Available The aim of this note was to record for the first time the finding of Hemilucilia segmentaria acting as biological vector of Dermatobia hominis, during a study of the diversity of Calliphoridae at Reserva Biológica do Tinguá, Rio de Janeiro, Brazil. The insects were captured using traps baited with chicken vicera, for a period of 28-30 h twice per month. In the period of one year, 1987 insects were collected, 7.5% of which belonged to the H. segmentaria; of these a female was captured in May 2001, carrying a mass of 20 eggs on the left side of its abdomen.

An In Vitro Evaluation of Ozonized Organic Extra-Virgin Olive Oil on Giardia Lamblia Cysts.

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Boland-Nazar, Najmeh Sadat; Eslamirad, Zahra; Sarmadian, Hossein; Ghasemikhah, Reza

2016-11-01

Giardia lamblia is a common intestinal parasite that has been reported all over the world. This study was conducted to evaluate the effect of ozonized organic extra-virgin olive oil on the cyst of G. lamblia . The olive oil was ozonized based on international standards and confirmed by the world health organization (WHO) at various times in a generator. The ozone concentration of olive oil was adjusted at 32, 64, 96, 128, 160 mg/g based on ozone absorption. Giardia lamblia cysts were isolated from heavily infected stool samples and the sucrose gradient flotation technique. Five groups of triple tubes containing Giardia cysts were exposed to olive oil with 32, 64, 96, 128, 160 ozone concentrations, and the sixth and seventh groups were exposed to non-ozonized olive oil and normal saline, respectively. The tubes were placed at room temperature, and every four hours, the mortality of the Giardia cysts was assessed. The results showed that the first five groups' mortality rate of Giardia cysts reached 100% in 100 hours. An increasing concentration of ozone in olive oil leads to an increase in the mortality rate of Giardia cysts. The results showed a significant difference in the mean time of the mortality in all the groups (P ≤ 0.05). Furthermore, the higher fatality effect of ozonized organic extra-virgin olive oil (Ozonized Olive Oil = OZO) was proved in comparison with metronidazole in vitro. We concluded that ozonized organic extra-virgin olive oil was a growth inhibitor of Giardia cysts, and concerning its compatibility with a biological system, it is recommended for further clinical trials.

Enhancement of anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array and Box-Behnken design.

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Khusro, Ameer; Aarti, Chirom; Dusthackeer, Azger; Agastian, Paul

2018-04-14

The prime focus of the present investigation was to optimize statistically the anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array (OA) and Box-Behnken design (BBD). The anti-tubercular activity of strain MANF2 was determined against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay. Among varied media examined, the isolate exhibited impressive anti-tubercular activity with paramount relative light unit reduction of >90% in de Man Rogose Sharpe (MRS) broth. Primarily, the anti-tubercular activity and biomass of strain MANF2 were estimated in MRS broth by optimizing eight diversified parameters using one factor at a time (OFAT) method after working out a series of experiments. The most significant contributing factors selected through OFAT tool were optimized using Taguchi approach with a standard OA layout of L18 (2 2  × 3 6 ). Results demonstrated the significant (P ≤ 0.05) influence of pH, temperature, yeast extract, magnesium sulphate, and glycerol on response variables. These controlled variables were further optimized using BBD matrix at N = 46 by second-order polynomial equation. The fermentation medium of pH 6.5 constituting yeast extract (0.5% w/v), magnesium sulphate (0.1% w/v), and glycerol (1.5% v/v), being further incubated at 30 °C showed enhanced anti-tubercular activity (98.7%) and approximately 4 fold increment in the bacterial biomass yield (8.3 mg/mL) with respect to traditional OFAT method. Three-dimensional response plots of the quadratic model showed interdependent interaction between the significant variables. In conclusion, the present study revealed the first report on the optimization of anti-tubercular activity and biomass of S. hominis via Taguchi OA as well as BBD design, and thus, paved a path for its proficient applications in pharmaceutical industries as dynamic mycobactericidal agent in future. Copyright © 2018

Collagen organization regulates stretch-initiated pain-related neuronal signals in vitro: Implications for structure-function relationships in innervated ligaments.

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Zhang, Sijia; Singh, Sagar; Winkelstein, Beth A

2018-02-01

Injury to the spinal facet capsule, an innervated ligament with heterogeneous collagen organization, produces pain. Although mechanical facet joint trauma activates embedded afferents, it is unclear if, and how, the varied extracellular microstructure of its ligament affects sensory transduction for pain from mechanical inputs. To investigate the effects of macroscopic deformations on afferents in collagen matrices with different organizations, an in vitro neuron-collagen construct (NCC) model was used. NCCs with either randomly organized or parallel aligned collagen fibers were used to mimic the varied microstructure in the facet capsular ligament. Embryonic rat dorsal root ganglia (DRG) were encapsulated in the NCCs; axonal outgrowth was uniform and in all directions in random NCCs, but parallel in aligned NCCs. NCCs underwent uniaxial stretch (0.25 ± 0.06 strain) corresponding to sub-failure facet capsule strains that induce pain. Macroscopic NCC mechanics were measured and axonal expression of phosphorylated extracellular signal-regulated kinase (pERK) and the neurotransmitter substance P (SP) was assayed at 1 day to assess neuronal activation and nociception. Stretch significantly upregulated pERK expression in both random and aligned gels (p organization. These findings suggest that collagen organization differentially modulates pain-related neuronal signaling and support structural heterogeneity of ligament tissue as mediating sensory function. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:770-777, 2018. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

the chentical composition and in vitro digestibility of forage samples ...

African Journals Online (AJOL)

oesophageal fistulated sheep, are presented in Table l. Table I. The average chemicol composition and in vitro digestibility offorage samples selectecl by oesophageal fistuhted sheep duing dffirent months of the yeor on noturol pasture. Month. Acid de- Lignin. Nitrogen tergent fibre. Cellulose In vitro digest- ibility of organ-.

Disorganized collagen scaffold interferes with fibroblast mediated deposition of organized extracellular matrix in vitro.

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Saeidi, Nima; Guo, Xiaoqing; Hutcheon, Audrey E K; Sander, Edward A; Bale, Shyam Sundar; Melotti, Suzanna A; Zieske, James D; Trinkaus-Randall, Vickery; Ruberti, Jeffrey W

2012-10-01

Many tissue engineering applications require the remodeling of a degradable scaffold either in vitro or in situ. Although inefficient remodeling or failure to fully remodel the temporary matrix can result in a poor clinical outcome, very few investigations have examined in detail, the interaction of regenerative cells with temporary scaffoldings. In a recent series of investigations, randomly oriented collagen gels were directly implanted into human corneal pockets and followed for 24 months. The resulting remodeling response exhibited a high degree of variability which likely reflects differing regenerative/synthetic capacity across patients. Given this variability, we hypothesize that a disorganized, degradable provisional scaffold could be disruptive to a uniform, organized reconstruction of stromal matrix. In this investigation, two established corneal stroma tissue engineering culture systems (collagen scaffold-based and scaffold-free) were compared to determine if the presence of the disorganized collagen gel influenced matrix production and organizational control exerted by primary human corneal fibroblast cells (PHCFCs). PHCFCs were cultured on thin disorganized reconstituted collagen substrate (RCS--five donors: average age 34.4) or on a bare polycarbonate membrane (five donors: average age 32.4 controls). The organization and morphology of the two culture systems were compared over the long-term at 4, 8, and 11/12 weeks. Construct thickness and extracellular matrix organization/alignment was tracked optically with bright field and differential interference contrast (DIC) microscopy. The details of cell/matrix morphology and cell/matrix interaction were examined with standard transmission, cuprolinic blue and quick-freeze/deep-etch electron microscopy. Both the scaffold-free and the collagen-based scaffold cultures produced organized arrays of collagen fibrils. However, at all time points, the amount of organized cell-derived matrix in the scaffold

Advances and perspectives in in vitro human gut fermentation modeling.

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Payne, Amanda N; Zihler, Annina; Chassard, Christophe; Lacroix, Christophe

2012-01-01

The gut microbiota is a highly specialized organ containing host-specific assemblages of microbes whereby metabolic activity directly impacts human health and disease. In vitro gut fermentation models present an unmatched opportunity of performing studies frequently challenged in humans and animals owing to ethical concerns. Multidisciplinary systems biology analyses supported by '-omics' platforms remain widely neglected in the field of in vitro gut fermentation modeling but are key to advancing the significance of these models. Model-driven experimentation using a combination of in vitro gut fermentation and in vitro human cell models represent an advanced approach in identifying complex host-microbe interactions and niches central to gut fermentation processes. The aim of this review is to highlight the advances and challenges exhibited by in vitro human gut fermentation modeling. Copyright © 2011 Elsevier Ltd. All rights reserved.

Organ In Vitro Culture: What Have We Learned about Early Kidney Development?

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Aleksandra Rak-Raszewska

2015-01-01

Full Text Available When Clifford Grobstein set out to study the inductive interaction between tissues in the developing embryo, he developed a method that remained important for the study of renal development until now. From the late 1950s on, in vitro cultivation of the metanephric kidney became a standard method. It provided an artificial environment that served as an open platform to study organogenesis. This review provides an introduction to the technique of organ culture, describes how the Grobstein assay and its variants have been used to study aspects of mesenchymal induction, and describes the search for natural and chemical inducers of the metanephric mesenchyme. The review also focuses on renal development, starting with ectopic budding of the ureteric bud, ureteric bud branching, and the generation of the nephron and presents the search for stem cells and renal progenitor cells that contribute to specific structures and tissues during renal development. It also presents the current use of Grobstein assay and its modifications in regenerative medicine and tissue engineering today. Together, this review highlights the importance of ex vivo kidney studies as a way to acquire new knowledge, which in the future can and will be implemented for developmental biology and regenerative medicine applications.

In vitro inactivation of Chlamydia trachomatis and of a panel of DNA (HSV-2, CMV, adenovirus, BK virus) and RNA (RSV, enterovirus) viruses by the spermicide benzalkonium chloride.

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Bélec, L; Tevi-Benissan, C; Bianchi, A; Cotigny, S; Beumont-Mauviel, M; Si-Mohamed, A; Malkin, J E

2000-11-01

Kinetics of inactivation by the detergent spermicide benzalkonium chloride (BZK) of Chlamydia trachomatis and of a panel of DNA viruses [herpes simplex virus hominis type 2 (HSV-2), cytomegalovirus (CMV), adenovirus (ADV) and BK virus (BKV)] and RNA [respiratory syncytial virus (RSV) and enterovirus (ENV)] were established in accordance with a standardized in vitro protocol. After a 5 min incubation, inactivation of >95% of HSV-2 and CMV was obtained at a concentration of 0.0025% (w/v) (25 Ig/L); concentrations as low as 0.0005%, 0.0050% and 0.0125%, induced a 3.0 log10 reduction in infectivity of HSV-2 and CMV, RSV and ADV, respectively. After a 60 min incubation, concentrations of 0.0125% and 0.050% provided a 3.0 log10 reduction in infectivity of ENV and BKV, respectively. These features indicate that sensitivity to BZK was very high (HSV-2 and CMV) or high (RSV) for enveloped viruses, intermediate (ADV) or low (ENV and BKV) for non-enveloped viruses. Furthermore, BZK had marked antichlamydial activity, showing >99% killing after only a 1 min incubation at a concentration of 0.00125%. BZK demonstrates potent in vitro activity against the majority of microorganisms causing sexually transmitted infectious diseases, including those acting as major genital cofactors of human immunodeficiency virus transmission. These attributes qualify BZK as a particularly attractive candidate for microbicide development.

Engineering stromal-epithelial interactions in vitro for ...

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Background: Crosstalk between epithelial and stromal cells drives the morphogenesis of ectodermal organs during development and promotes normal mature adult epithelial tissue function. Epithelial-mesenchymal interactions (EMIs) have been examined using mammalian models, ex vivo tissue recombination, and in vitro co-cultures. Although these approaches have elucidated signaling mechanisms underlying morphogenetic processes and adult mammalian epithelial tissue function, they are limited by the availability of human tissue, low throughput, and human developmental or physiological relevance. Objectives: Bioengineering strategies to promote EMIs using human epithelial and mesenchymal cells have enabled the development of human in vitro models of adult epidermal and glandular tissues. In this review, we describe recent bioengineered models of human epithelial tissue and organs that can instruct the design of organotypic models of human developmental processes.Methods: We reviewed current bioengineering literature and here describe how bioengineered EMIs have enabled the development of human in vitro epithelial tissue models.Discussion: Engineered models to promote EMIs have recapitulated the architecture, phenotype, and function of adult human epithelial tissue, and similar engineering principles could be used to develop models of developmental morphogenesis. We describe how bioengineering strategies including bioprinting and spheroid culture could be implemented to

In vitro effects of 5-hydroxytryptophan, indoleamines and leptin on arylalkylamine N-acetyltransferase (AA-NAT) activity in pineal organ of the fish, Clarias gariepinus (Burchell, 1822) during different phases of the breeding cycle.

Science.gov (United States)

Gupta, B B P; Yanthan, L; Singh, Ksh Manisana

2010-08-01

Arylalkylamine N-acetyltransferase (AA-NAT) is the rate-limiting enzyme of melatonin biosynthetic pathway. In vitro effects of 5-hydroxytryptophan (5-HTP) and indoleamines (serotonin, N-acetylserotonin and melatonin) were studied on AA-NAT activity in the pineal organ of the fish, C. gariepinus during different phases of its annual breeding cycle. Further, in vitro effects of leptin on AA-NAT activity in the pineal organ were studied in fed and fasted fishes during summer and winter seasons. Treatments with 5-HTP and indoleamines invariably stimulated pineal AA-NAT activity in a dose-dependent manner during all the phases. However, leptin increased AA-NAT activity in a dose-dependent manner only in the pineal organ of the fed fishes, but not of the fasted fishes irrespective of the seasons.

In vitro organic matter digestibility and gas production of fish-meal ...

African Journals Online (AJOL)

user

2011-03-28

Mar 28, 2011 ... In this study, an in vitro rumen gas production technique was utilized to evaluate fish-meal coated with ... Keywords: fish-meal; gas production; hydrogenated tallow; .... industrial city, Saveh, Iran). ..... commercial dairy rations.

In vitro techniques for crop improvement

International Nuclear Information System (INIS)

1985-01-01

The film refers to principles of plant tissue culture - laboratory requirements, media preparation, explant establishment and subculturing method. In vitro growth and development of crop plants are demonstrate and the application of in vitro techniques in plant breeding is discussed. The second part of the film shows the application of cell, tissue and organ culture in plants. Micropropagation and virus eradication are important technologies for the improvement of vegetatively propagated plants; zygotic embryo rescue techniques are used for distant hybridization, especially in cereals. Plant biotechnology offers a potent means for the in vitro generation of enhanced genetic variability - somaclonal and mutagen induced variation. Principles of the isolation and culture of plant protoplasts are explained and their potential for somatic hybridization in higher plants is demonstrated. Haploids are valuable to accelerate breeding cycles of plants. Plant biotechnology is described as an important complementary tool to conventional plant breeding methods

Leukocyte Lysis and Cytokine Induction by the Human Sexually Transmitted Parasite Trichomonas vaginalis.

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Frances Mercer

2016-08-01

Full Text Available Trichomonas vaginalis (Tv is an extracellular protozoan parasite that causes the most common non-viral sexually transmitted infection: trichomoniasis. While acute symptoms in women may include vaginitis, infections are often asymptomatic, but can persist and are associated with medical complications including increased HIV susceptibility, infertility, pre-term labor, and higher incidence of cervical cancer. Heightened inflammation resulting from Tv infection could account for these complications. Effective cellular immune responses to Tv have not been characterized, and re-infection is common, suggesting a dysfunctional adaptive immune response. Using primary human leukocyte components, we have established an in vitro co-culture system to assess the interaction between Tv and the cells of the human immune system. We determined that in vitro, Tv is able to lyse T-cells and B-cells, showing a preference for B-cells. We also found that Tv lysis of lymphocytes was mediated by contact-dependent and soluble factors. Tv lysis of monocytes is far less efficient, and almost entirely contact-dependent. Interestingly, a common symbiont of Tv, Mycoplasma hominis, did not affect cytolytic activity of the parasite, but had a major impact on cytokine responses. M. hominis enabled more diverse inflammatory cytokine secretion in response to Tv and, of the cytokines tested, Tv strains cleared of M. hominis induced only IL-8 secretion from monocytes. The quality of the adaptive immune response to Tv is therefore likely influenced by Tv symbionts, commensals, and concomitant infections, and may be further complicated by direct parasite lysis of effector immune cells.

Leukocyte Lysis and Cytokine Induction by the Human Sexually Transmitted Parasite Trichomonas vaginalis

Science.gov (United States)

Mercer, Frances; Diala, Fitz Gerald I.; Chen, Yi-Pei; Molgora, Brenda M.; Ng, Shek Hang; Johnson, Patricia J.

2016-01-01

Trichomonas vaginalis (Tv) is an extracellular protozoan parasite that causes the most common non-viral sexually transmitted infection: trichomoniasis. While acute symptoms in women may include vaginitis, infections are often asymptomatic, but can persist and are associated with medical complications including increased HIV susceptibility, infertility, pre-term labor, and higher incidence of cervical cancer. Heightened inflammation resulting from Tv infection could account for these complications. Effective cellular immune responses to Tv have not been characterized, and re-infection is common, suggesting a dysfunctional adaptive immune response. Using primary human leukocyte components, we have established an in vitro co-culture system to assess the interaction between Tv and the cells of the human immune system. We determined that in vitro, Tv is able to lyse T-cells and B-cells, showing a preference for B-cells. We also found that Tv lysis of lymphocytes was mediated by contact-dependent and soluble factors. Tv lysis of monocytes is far less efficient, and almost entirely contact-dependent. Interestingly, a common symbiont of Tv, Mycoplasma hominis, did not affect cytolytic activity of the parasite, but had a major impact on cytokine responses. M. hominis enabled more diverse inflammatory cytokine secretion in response to Tv and, of the cytokines tested, Tv strains cleared of M. hominis induced only IL-8 secretion from monocytes. The quality of the adaptive immune response to Tv is therefore likely influenced by Tv symbionts, commensals, and concomitant infections, and may be further complicated by direct parasite lysis of effector immune cells. PMID:27529696

Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity.

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Baudoin, Régis; Corlu, Anne; Griscom, Laurent; Legallais, Cécile; Leclerc, Eric

2007-06-01

Current developments in the technological fields of liver tissue engineering, bioengineering, biomechanics, microfabrication and microfluidics have lead to highly complex and pertinent new tools called "cell biochips" for in vitro toxicology. The purpose of "cell biochips" is to mimic organ tissues in vitro in order to partially reduce the amount of in vivo testing. These "cell biochips" consist of microchambers containing engineered tissue and living cell cultures interconnected by a microfluidic network, which allows the control of microfluidic flows for dynamic cultures, by continuous feeding of nutrients to cultured cells and waste removal. Cell biochips also allow the control of physiological contact times of diluted molecules with the tissues and cells, for rapid testing of sample preparations or specific addressing. Cell biochips can be situated between in vitro and in vivo testing. These types of systems can enhance functionality of cells by mimicking the tissue architecture complexities when compared to in vitro analysis but at the same time present a more rapid and simple process when compared to in vivo testing procedures. In this paper, we first introduce the concepts of microfluidic and biochip systems based on recent progress in microfabrication techniques used to mimic liver tissue in vitro. This includes progress and understanding in biomaterials science (cell culture substrate), biomechanics (dynamic cultures conditions) and biology (tissue engineering). The development of new "cell biochips" for chronic toxicology analysis of engineered tissues can be achieved through the combination of these research domains. Combining these advanced research domains, we then present "cell biochips" that allow liver chronic toxicity analysis in vitro on engineered tissues. An extension of the "cell biochip" idea has also allowed "organ interactions on chip", which can be considered as a first step towards the replacement of animal testing using a combined liver

Regeneration of some monocotyledonous plants from subterranean organs in vitro

OpenAIRE

Krystyna D. Kromer

2013-01-01

The aim in view was investigation ofthe regenerative potential of rhizomes, bulbs and corms of ten monocotyledonous plant species from four families: Amaryllidaceae (Haemanthus katharinae, Crinum abyssinicum, Leucojum vernum), Araceae (Spathiphyllum wallisii), Iridaceae (Crocus vernus, Iris germanica), Liliaceae (Hosta lancifolia, Muscari racemosum, Scilla laxiflora, Veltheimia viridifolia) under conditions of in vitro culture. All the investigated species were capable of buld or, alternative...

Influence of different cytokinins used in in vitro culture on the stoma ...

African Journals Online (AJOL)

GREGO

2007-03-05

Mar 5, 2007 ... In vitro micropropagation of Pistachio (Pistacia vera var. Siirt), Plant Cell. Tissue Organ Cult. 77: 215–219. Pospisilova J, Ticha I, Kadlecek P, Haisel D, Plzakova S (1990a). Acclimatization of micropropagated plants to ex vitro conditions. Biologia Plantarum, 42: 481–497. Salarova J, Pospisilova J (1997).

Associated microbial contaminants in in-vitro micropropagation of ...

African Journals Online (AJOL)

Studies were carried out to determine the microbial contaminants associated with in-vitro micropropagation of Ipomea batatas (sweet potato). The contaminants were found to be mostly fungal organisms, Aspergillus Spp (62%), Penicillum Spp. (31%), Fusarium Spp. (5%) and Alternaria Spp. (2%). Bacterial contamination ...

3D in vitro technology for drug discovery.

Science.gov (United States)

Hosseinkhani, Hossein

2012-02-01

Three-dimensional (3D) in vitro systems that can mimic organ and tissue structure and function in vivo, will be of great benefit for a variety of biological applications from basic biology to toxicity testing and drug discovery. There have been several attempts to generate 3D tissue models but most of these models require costly equipment, and the most serious disadvantage in them is that they are too far from the mature human organs in vivo. Because of these problems, research and development in drug discovery, toxicity testing and biotech industries are highly expensive, and involve sacrifice of countless animals and it takes several years to bring a single drug/product to the market or to find the toxicity or otherwise of chemical entities. Our group has been actively working on several alternative models by merging biomaterials science, nanotechnology and biological principles to generate 3D in vitro living organs, to be called "Human Organs-on-Chip", to mimic natural organ/tissues, in order to reduce animal testing and clinical trials. We have fabricated a novel type of mechanically and biologically bio-mimicking collagen-based hydrogel that would provide for interconnected mini-wells in which 3D cell/organ culture of human samples in a manner similar to human organs with extracellular matrix (ECM) molecules would be possible. These products mimic the physical, chemical, and biological properties of natural organs and tissues at different scales. This paper will review the outcome of our several experiments so far in this direction and the future perspectives.

Estudio comparativo de la frecuencia de Blastocystis hominis en niños en edad preescolar de una zona urbana y una rural de la ciudad de Cartagena de Indias y su relación con las manifestaciones clínicas y factores de riesgo

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Mavianis Pinilla Pérez

2015-01-01

Full Text Available Introducción: las parasitosis intestinales son infecciones comunes a nivel mundial y de mayor prevalencia en las comunidades empobrecidas de países en desarrollo. A finales de los ochenta el Blastocystis hominis no era reportado rutinariamente, representando solo un problema de diagnóstico diferencial; pero en los noventa mediante un oportuno diagnóstico aumenta su prevalencia de 1,5 a 10% en los países desarrollados y de 30 a 50 % en países en desarrollo. Objetivo: comparar la frecuencia de Blastocystis hominis en niños de preescolar de una zona urbana (Crespo y una rural (Boquilla de Cartagena Bolívar y su relación con las manifestaciones clínicas, factores de riesgo. Materiales y métodos: se analizaron mediante la técnica de coprológico seriado, 286 muestras de materia fecal de niños de 1 a 5 años que procedían de una zona urbana y una zona rural de la ciudad de Cartagena de indias. Resultados: se encontraron parásitos en el 38,5% (55/143 de los niños de la zona urbana y el 79% (113/143 de la zona rural. Se encontró asociación entre la presencia de parásitos con la procedencia de los niños evaluados (X2= 48,53; p<0,0001. Se encontró una frecuencia de 51,1% de B. hominis en la zona urbana y 44,8% en la zona rural. Conclusiones: la falta de seguimiento y control de los casos problema sugiere la necesidad de formular e implementar medidas orientadas a mejorar la calidad de vida en las zonas estudiadas.

Explaining an Unusually Fast Parasitic Enzyme: Folate Tail-Binding Residues Dictate Substrate Positioning and Catalysis in Cryptosporidium hominis Thymidylate Synthase

Energy Technology Data Exchange (ETDEWEB)

Martucci,W.; Vargo, M.; Anderson, K.

2008-01-01

The essential enzyme TS-DHFR from Cryptosporidium hominis undergoes an unusually rapid rate of catalysis at the conserved TS domain, facilitated by two nonconserved residues, Ala287 and Ser290, in the folate tail-binding region. Mutation of these two residues to their conserved counterparts drastically affects multiple steps of the TS catalytic cycle. We have determined the crystal structures of all three mutants (A287F, S290G, and A287F/S290G) in complex with active site ligands dUMP and CB3717. The structural data show two effects of the mutations: an increased distance between the ligands in the active site and increased flexibility of the folate ligand in the partially open enzyme state that precedes conformational change to the active catalytic state. The latter effect is able to be rescued by the mutants containing the A287F mutation. In addition, the conserved water network of TS is altered in each of the mutants. The structural results point to a role of the folate tail-binding residues in closely positioning ChTS ligands and restricting ligand flexibility in the partially open state to allow for a rapid transition to the active closed state and enhanced rate of catalysis. These results provide an explanation on how folate tail-binding residues at one end of the active site affect long-range interactions throughout the TS active site and validate these residues as targets for species-specific drug design.

Organic Additives Improves the in Vitro Growth of Native Orchid Vanda helvola Blume

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Devina DAVID

2015-06-01

Full Text Available In vitro seed germination has been proven to be the most efficient technique to propagate orchid. The application of this aseptic technique has contributed to conservation of many endangered orchid species. In this study, undehisced capsules of Vanda helvola Blume were collected from Orchid Conservation Centre in Lagud Sebrang Agriculture Park, after 120 days from hand pollination and aseptically cultured on three types of basal media such as Murashige and Skoog (MS, Knudson C (KC and Vacin and Went (VW. After 90 days of culture, 66.40 ± 4.14% of seeds successfully germinated on KC medium. The effect of organic additives such as tomato juice, coconut water, peptone and yeast extract at different level of concentrations in KC basal medium were also tested on seed germination and seedling development of this native orchid. After 90 days of culture, over 90% of seeds were tremendously germinated on KC medium supplemented with 10% or 15% (v/v of tomato juice. The incorporation of peptone at 0.1% (w/v in KC basal media promoted rapid development of protocorm to seedling. Seedlings on this treatment produced an average of three leaves and two roots after 90 days of culture and were successfully acclimatized.

Ribonucleoprotein organization of eukaryotic RNA. XXXII. U2 small nuclear RNA precursors and their accurate 3' processing in vitro as ribonucleoprotein particles.

Science.gov (United States)

Wieben, E D; Nenninger, J M; Pederson, T

1985-05-05

Biosynthetic precursors of U2 small nuclear RNA have been identified in cultured human cells by hybrid-selection of pulse-labeled RNA with cloned U2 DNA. These precursor molecules are one to approximately 16 nucleotides longer than mature U2 RNA and contain 2,2,7-trimethylguanosine "caps". The U2 RNA precursors are associated with proteins that react with a monoclonal antibody for antigens characteristic of small nuclear ribonucleoprotein particles. Like previously described precursors of U1 and U4 small nuclear RNAs, the pre-U2 RNAs are recovered in cytoplasmic fractions, although it is not known if this is their location in vivo. The precursors are processed to mature-size U2 RNA when cytoplasmic extracts are incubated in vitro at 37 degrees C. Mg2+ is required but ATP is not. The ribonucleoprotein structure of the pre-U2 RNA is maintained during the processing reaction in vitro, as are the 2,2,7-trimethylguanosine caps. The ribonucleoprotein organization is of major importance, as exogenous, protein-free U2 RNA precursors are degraded rapidly in the in vitro system. Two lines of evidence indicate that the conversion of U2 precursors to mature-size U2 RNA involves a 3' processing reaction. First, the reaction is unaffected by a large excess of mature U2 small nuclear RNP, whose 5' trimethylguanosine caps would be expected to compete for a 5' processing activity. Second, when pre-U2 RNA precursors are first stoichiometrically decorated with an antibody specific for 2,2,7-trimethylguanosine, the extent of subsequent processing in vitro is unaffected. These results provide the first demonstration of a eukaryotic RNA processing reaction in vitro occurring within a ribonucleoprotein particle.

Effects of caffeine on circadian phase, amplitude and period evaluated in cells in vitro and peripheral organs in vivo in PER2::LUCIFERASE mice

Science.gov (United States)

Narishige, Seira; Kuwahara, Mari; Shinozaki, Ayako; Okada, Satoshi; Ikeda, Yuko; Kamagata, Mayo; Tahara, Yu; Shibata, Shigenobu

2014-01-01

Background and Purpose Caffeine is one of the most commonly used psychoactive substances. Circadian rhythms consist of the main suprachiasmatic nucleus (SCN) clocks and peripheral clocks. Although caffeine lengthens circadian rhythms and modifies phase changes in SCN-operated rhythms, the effects on caffeine on the phase, period and amplitude of peripheral organ clocks are not known. In addition, the role of cAMP/Ca2+ signalling in effects of caffeine on rhythm has not been fully elucidated. Experimental Approach We examined whether chronic or transient application of caffeine affects circadian period/amplitude and phase by evaluating bioluminescence rhythm in PER2::LUCIFERASE knock-in mice. Circadian rhythms were monitored in vitro using fibroblasts and ex vivo and in vivo for monitoring of peripheral clocks. Key Results Chronic application of caffeine (0.1–10 mM) increased period and amplitude in vitro. Transient application of caffeine (10 mM) near the bottom of the decreasing phase of bioluminescence rhythm caused phase advance in vitro. Caffeine (0.1%) intake caused a phase delay under light–dark or constant dark conditions, suggesting a period-lengthening effect in vivo. Caffeine (20 mg·kg−1) at daytime or at late night-time caused phase advance or delay in bioluminescence rhythm in the liver and kidney respectively. The complicated roles of cAMP/Ca2+ signalling may be involved in the caffeine-induced increase of period and amplitude in vitro. Conclusions and Implications Caffeine affects circadian rhythm in mice by lengthening the period and causing a phase shift of peripheral clocks. These results suggest that caffeine intake with food/drink may help with food-induced resetting of peripheral circadian clocks. PMID:25160990

A Study of Associated Factors, Including Genital Herpes, in Black ...

African Journals Online (AJOL)

Herpesvirus hominis antibodies were measured by a kinetic neutralisation test. Among Black patients in the Johannesburg area there is a high incidence of carcinoma of the cervix, and H. hominis type 2 infection. A correlation exists between carcinomas of the cervix on the one hand and antibodies to H. hominis type 2 on ...

Novel In Vitro/Ex Vivo Animal Modeling for Filovirus Aerosol Infection

Science.gov (United States)

2014-09-01

Infection PRINCIPAL INVESTIGATOR: Ayesha Mahmood, Ph.D. CONTRACTING ORGANIZATION: Sanofi Pasteur VaxDesign Corporation...ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORT Sanofi Pasteur VaxDesign Corporation Orlando, Florida, 32826 9...a collaborative research effort between the USAMRIID Labs and Sanofi Pasteur VaxDesign to develop in vitro and ex vivo viral disease model systems

Enzymatically Degradable Hybrid Organic-Inorganic Bridged Silsesquioxane Nanoparticles for In-Vitro Imaging

KAUST Repository

Fatieiev, Yevhen

2015-06-30

Non-aggregated dense bridged silsesquioxane (BS) nanoparticles based on nature-inspired oxamide bridges are shown to degrade in simulated biological media upon cleavage with endopeptidase. Fluorescent BS nanoprobes with incorporated fluorescein dyes were applied for in-vitro imaging in cancer cells.

Cytogenetic characterization of Encyclia caximboensis cultivated in vitro (Orchidaceae)

OpenAIRE

Gizelly Mendes Silva; Tatiane Lemos Varella; Kaliane Maximiliano Cruz; Ilio Fealho Carvalho; Isane Vera Karsburg; Maurecilne Lemes Silva

2015-01-01

Encyclia caximboensis is an Amazonian species endemic to the Serra do Cachimbo, which is located between the northern of the Mato Grosso state and the southern part of Para state. Studies reporting in vitro cultivation and cytogenetic characterization of this species are still scarce. Therefore, the objective of this work was to determine the cytogenetic characteristics and to identify the nucleolar organizer regions (NORs) of the species E. Caximboensis, cultivated in vitro. Seeds of E. caxi...

Associação entre presença de Mycoplasma hominis e Ureaplasma urealyticum e níveis de citocinas pró e antiinflamatórias no líquido amniótico de gestação de termo

OpenAIRE

Ramos, Bruna Ribeiro de Andrade [UNESP

2011-01-01

Microbial invasion of the amniotic cavity has been described in term deliveries and its role on the immune modulation is of interest to the better understanding of the underlying labor processes. The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum in the amniotic fluid of term pregnancies and to evaluate its influence on cytokines production at the end of pregnancy. A cross sectional study was conducted with fifty five pregnant women out of l...

In vitro sensitivity of Trichomonas vaginalis and Candida albicans to chemotherapeutic agents.

Science.gov (United States)

Lövgren, T; Salmela, I

1978-06-01

Strains of fresh clinical isolates of Trichomonas vaginalis and Candida albicans have been tested in vitro for their sensitivity to eight drugs used in the therapy of monilial and trichomonal vaginitis. Three of the chemotherapeutic agents, chlorchinaldol, clotrimazole and broxyquinoline were effective against both organisms. Tinidazole and metronidazole were active against T. vaginalis. The strains of C. albicans were also sensitive to trichomycin, natamycin and nystatin. Tinidazole was the most effective trichomonacide, clotrimazole and chlorchinaldol were most effective against C. albicans, while chlorchinaldol had the best in vitro effect against both organisms. The ranges of the MICs are compared to values previously reported.

In vitro activity of tigecycline and comparator agents against a global collection of Gram-negative and Gram-positive organisms: tigecycline Evaluation and Surveillance Trial 2004 to 2007.

Science.gov (United States)

Garrison, Mark W; Mutters, Reinier; Dowzicky, Michael J

2009-11-01

The Tigecycline Evaluation and Surveillance Trial began in 2004 to monitor the in vitro activity of tigecycline and comparator agents against a global collection of Gram-negative and Gram-positive pathogens. Against Gram negatives (n = 63 699), tigecycline MIC(90)'s ranged from 0.25 to 2 mg/L for Escherichia coli, Haemophilus influenzae, Acinetobacter baumannii, Klebsiella oxytoca, Enterobacter cloacae, Klebsiella pneumoniae, and Serratia marcescens (but was > or =32 for Pseudomonas aeruginosa). Against Gram-positive organisms (n = 32 218), tigecycline MIC(90)'s were between 0.06 and 0.25 mg/L for Streptococcus pneumoniae, Enterococcus faecium, Streptococcus agalactiae, Staphylococcus aureus, and Enterococcus faecalis. The in vitro activity of tigecycline was maintained against resistant phenotypes, including multidrug-resistant A. baumannii (9.2% of isolates), extended-spectrum beta-lactamase-producing E. coli (7.0%) and K. pneumoniae (14.0%), beta-lactamase-producing H. influenzae (22.2%), methicillin-resistant S. aureus (44.5%), vancomycin-resistant E. faecium (45.9%) and E. faecalis (2.8%), and penicillin-resistant S. pneumoniae (13.8%). Tigecycline represents a welcome addition to the armamentarium against difficult to treat organisms.

Self-organization of spatial patterning in human embryonic stem cells

Science.gov (United States)

Deglincerti, Alessia; Etoc, Fred; Ozair, M. Zeeshan; Brivanlou, Ali H.

2017-01-01

The developing embryo is a remarkable example of self-organization, where functional units are created in a complex spatio-temporal choreography. Recently, human embryonic stem cells (ESCs) have been used to recapitulate in vitro the self-organization programs that are executed in the embryo in vivo. This represents a unique opportunity to address self-organization in humans that is otherwise not addressable with current technologies. In this essay, we review the recent literature on self-organization of human ESCs, with a particular focus on two examples: formation of embryonic germ layers and neural rosettes. Intriguingly, both activation and elimination of TGFβ signaling can initiate self-organization, albeit with different molecular underpinnings. We discuss the mechanisms underlying the formation of these structures in vitro and explore future challenges in the field. PMID:26970615

Modelling the association between in vitro gas production and ...

African Journals Online (AJOL)

In vitro gas production of four different browse plants (Azadirachta indica, Terminalia catappa, Mangifera indica and Vernonia amygdalina) was investigated under different extractions. The relationship between the forage composition parameters (dry matter, organic matter, crude protein, acid detergent fibre, neutral ...

Babesia bigemina: in vitro cultivation and characterization

International Nuclear Information System (INIS)

Vega Y Murguia, C.A.

1985-01-01

An in vitro model for the continuous replication of Babesia bigemina was developed and this model was used to study the parasite's biology. Initially, infected erythrocytes from a calf inoculated with a strain of B. bigemina was suspended with normal bovine erythrocytes and the parasite propagated in vitro. The cultured organism was inoculated into another calf and reproduced the disease with typical signs. Babesia bigemina was reisolated in pure culture in vitro. The animal recovered after receiving specific treatment. A procedure was developed to cryopreserve infected erythrocytes and merozoites to initiate in vitro cultures. Homogeneous parasite populations were obtained by cloning by limiting dilution. Parasitic growth was detected between 16-28 days after dilutions were made. Three primary clones were selected for recloning. Infected erythrocytes from the original isolate nd the clones were concentrated by Percoll density gradients. Density values for paired and single infected cells were determined. Enzymatic content of concentrated infected cells was analyzed by starch gel electrophoresis. Enzymes LDH, GPI, and GDH were detected, but polymorphism among clones was not observed. The enzyme 6-PDG was not associated with the parasite. Separation of labelled proteins was done by SDS-PAGE. The separation patterns were similar for all samples. a 43 Kd polypeptide was detected in the B. bigemina culture supernatant

Prevalence, risk factors of infection and molecular characterization of trichomonads in puppies from French breeding kennels.

Science.gov (United States)

Grellet, Aurélien; Brunopolack; Feugier, Alexandre; Boucraut-Baralon, Corine; Grandjean, Dominique; Vandewynckel, Laurine; Cian, Amandine; Meloni, Dionigia; Viscogliosi, Eric

2013-11-08

The trichomonad species Tritrichomonas fetus and Pentatrichomonas hominis were recently identified in the feces of dogs with diarrhea. However the prevalence and pathogenicity of these parasites in the canine population still remained poorly resolved. Therefore the aim of the present study was (1) to determine the prevalence of trichomonads infecting puppies living in French breeding kennels, (2) to confirm the predominance of P. hominis in dogs, (3) to investigate the genetic diversity of P. hominis isolates identified in the French canine population and (4) to evaluate the risk factors for infection by P. hominis and the influence of the parasite on feces consistency. A total of 215 both diarrheic and non-diarrheic puppies from 25 French breeding kennels were included in this epidemiological survey. Fecal samples from each puppy were examined for 6 gastrointestinal pathogens: parvovirus type 2 (CPV2), coronavirus, Toxocara canis, Cystoisospora ohioensis-complex, Cystoisospora canis, and Giardia intestinalis. A part of each collected stool was also tested for the presence of motile trichomonads by microscopy after culturing. The prevalence of trichomonad infection was 15.8% (34/215) among puppies and 20% (5/25) among breeding kennels. DNA from 26 of the 34 positive samples was successfully amplified using a trichomonad-specific primer pair. Analysis of the sequences of PCR products indicated that P. hominis was the only trichomonad infecting the canine population. All the puppies infected with P. hominis belonged to large breed dogs. Moreover, puppies from large breeding kennels, excreting a high level of G. intestinalis and/or excreting a high level of C. canis oocysts showed a higher probability of being positive for P. hominis infection. Univariate analysis also revealed an increased risk for P. hominis infection in puppies with abnormal feces. However, in a multivariate analysis, CPV2 was the only gastrointestinal pathogen associated with abnormal feces. Since

Effect of incorporation of walnut cake (Juglans regia in concentrate mixture on degradation of dry matter, organic matter and production of microbial biomass in vitro in goat

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Mohsin Ahmad Mir

2015-10-01

Full Text Available Aim: This study was carried out to investigate the effect of incorporation of different level of walnut cake in concentrate mixture on in vitro dry matter degradation in order to determine its level of supplementation in ruminant ration. Materials and Methods: Walnut cake was used @ 0, 10, 15, 20, 25 and 30% level to formulate an iso-nitrogenous concentrate mixtures and designated as T1, T2, T3, T4, T5 and T6 respectively. The different formulae of concentrate mixtures were used for in vitro gas production studies using goat rumen liquor with wheat straw in 40:60 ratio. Proximate composition, fiber fractionation and calcium and phosphrous content of walnut cake were estimated. Result: The per cent IVDMD value of T1 and T2 diets was 68.42 ± 1.20 and 67.25 ± 1.37 respectively which was found highest (P<0.05 T3, T4, T5 and T6. Similar trend was also found for TDOM and MBP. Inclusion of walnut cake at 10% level in the concentrate mixture does not affect in vitro dry matter digestibility (IVDMD, truly degradable organic matter (TDOM, mg/200 mg DM, total gas production, microbial biomass production (MBP and efficiency of microbial biomass production (EMP. Conclusion: It is concluded that walnut cake incorporation up to 10% level in the iso -nitrogenous concentrate mixture has no any negative effect on in vitro digestibility of dry matter (DM, TDOM, MBP, EMP and total gas production in goat.

Effects of mild ozonisation on gene expression and nuclear domains organization in vitro.

Science.gov (United States)

Scassellati, C; Costanzo, M; Cisterna, B; Nodari, A; Galiè, M; Cattaneo, A; Covi, V; Tabaracci, G; Bonvicini, C; Malatesta, M

2017-10-01

In the last two decades, the use of ozone (O 3 ) as a complementary medical approach has progressively been increasing; however, its application is still limited due to the numerous doubts about its possible toxicity, despite the low concentrations used in therapy. For an appropriate and safe clinical application of a potentially toxic agent such as O 3 , it is crucial to elucidate the cellular response to its administration. Molecular analyses and transmission electron microscopy were here combined to investigate in vitro the effects of O 3 administration on transcriptional activity and nuclear domains organization of cultured SH-SY5Y neuronal cells; low O 3 concentrations were used as those currently administered in clinical practice. Mild ozonisation did not affect cell proliferation or death, while molecular analyses showed an O 3 -induced modulation of some genes involved in the cell response to stress (HMOX1, ERCC4, CDKN1A) and in the transcription machinery (CTDSP1). Ultrastructural cytochemistry after experiments of bromouridine incorporation consistently demonstrated an increased transcriptional rate at both the nucleoplasmic (mRNA) and the nucleolar (rRNA) level. No ultrastructural alteration of nuclear domains was observed. Our molecular, ultrastructural and cytochemical data demonstrate that a mild toxic stimulus such as mild ozonisation stimulate cell protective pathways and nuclear transcription, without altering cell viability. This could possibly account for the positive effects observed in ozone-treated patients. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nitrogen metabolism in a grapevine in vitro system

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Nuria Llorens

2002-09-01

Full Text Available Ammonium, nitrate, nitrite, protein and individual and total amino acid contents were determined in grapevine (cv Sauvignon cultured in vitro. The enzyme activities of nitrate and nitrite reductases, glutamine synthetase, glutamate synthetase and dehydrogenase were also determined. The nitrogen taken up by the plants was 70% of the total nitrogen in the medium after 75 days of in vitro culture. Most of the nitrogen taken up was recovered in the leaves, yet only ammonia and amino acid concentrations were significantly higher in leaves. In roots, glutamine was the most abundant amino acid. In leaves, the most abundant amino acids were aspartate, glutamate, glutamine, alanine, arginine and g-aminobutirate. All enzyme activities were higher in roots than in leaves. These results suggest that both roots and leaves incorporate inorganic nitrogen into organic forms.

In Vitro Antibiotic Susceptibility Studies Of Bacteria Associated With ...

African Journals Online (AJOL)

In vitro tests of the susceptibility of isolates of bacterial keratitis pathogens to antibiotics were carried out in this study. Staphylococcus aureus was the most frequently isolated organisms followed by Pseudomonas aeruginosa and Streptococcus pneumoniae. Antibiotic sensitivity testing showed a high susceptibility to ...

In vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah.

Science.gov (United States)

Hossain, M Amzad; Shah, Muhammad Dawood; Gnanaraj, Charles; Iqbal, Muhammad

2011-09-01

To detect the in vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah. The dry powder leaves of Tetrastigma were extracted with different organic solvent such as hexane, ethyl acetate, chloroform, butanol and aqueous methanol. The total phenolic and total flavonoids contents of the essential oil and various organic extracts such as hexane, ethyl acetate, chloroform, butanol and aqueous ethanol were determined by Folin - Ciocalteu method and the assayed antioxidant activity was determined in vitro models such as antioxidant capacity by radical scavenging activity using α, α-diphenyl- β-picrylhydrazyl (DPPH) method. The total phenolic contents of the essential oil and different extracts as gallic acid equivalents were found to be highest in methanol extract (386.22 mg/g) followed by ethyl acetate (190.89 mg/g), chloroform (175.89 mg/g), hexane (173.44 mg/g), and butanol extract (131.72 mg/g) and the phenolic contents not detected in essential oil. The antioxidant capacity of the essential oil and different extracts as ascorbic acid standard was in the order of methanol extract > ethyl acetate extract >chloroform> butanol > hexane extract also the antioxidant activity was not detected in essential oil. The findings show that the extent of antioxidant activity of the essential oil and all extracts are in accordance with the amount of phenolics present in that extract. Leaves of Tetrastigma being rich in phenolics may provide a good source of antioxidant. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

The effect of Hominis Placenta Pharmacopuncture on Leg spasticity of stroke patients (A Pilot study, Double blind, Randomized, Controlled Clinical Trial

Directory of Open Access Journals (Sweden)

Ju-hwan Noh

2009-12-01

Full Text Available Objective : The purpose of this study is to determine the effect of Hominis Placenta Pharmacopuncture(HPP on lower limb spasticity control in stroke patients. Methods : Twenty stroke patients with Leg spasticity were randomly divided into two groups, a Distilled water Pharmacopuncture(group I and a HPP(group II. The number of Pharmacopuncture was 5 times a week and acupuncture treatment was 3 times a week for 3 weeks. Modified Ashworth Scale(MAS, H-reflex/M-response ratio(H/M ratio, Berg Balance Scale(BBS and Time Up & Go(TUG were used for evaluation of spasticity control before experiment, after 1 week, 2 weeks, 3 weeks. Results : Group I showed significant improvement(p<.05 in BBS but no significant improvement in MAS, H/M ratio, and TUG. Group II showed significant improvement(p<.05 in MAS, BBS, and TUG, but no significant improvement in H/M ratio. The results showed significant difference in TUG, but no significant difference in MAS, H/M ratio and BBS between 2 groups. Conclusion : These results showed that HPP might decrease lower limb spasticity and increase leg motor function in stroke patients. Further studies will be required to examine more cases in the long period for the effect on lower limb in spasticity by HPP.

Self-Organization of Spatial Patterning in Human Embryonic Stem Cells.

Science.gov (United States)

Deglincerti, Alessia; Etoc, Fred; Ozair, M Zeeshan; Brivanlou, Ali H

2016-01-01

The developing embryo is a remarkable example of self-organization, where functional units are created in a complex spatiotemporal choreography. Recently, human embryonic stem cells (ESCs) have been used to recapitulate in vitro the self-organization programs that are executed in the embryo in vivo. This represents an unique opportunity to address self-organization in humans that is otherwise not addressable with current technologies. In this chapter, we review the recent literature on self-organization of human ESCs, with a particular focus on two examples: formation of embryonic germ layers and neural rosettes. Intriguingly, both activation and elimination of TGFβ signaling can initiate self-organization, albeit with different molecular underpinnings. We discuss the mechanisms underlying the formation of these structures in vitro and explore future challenges in the field. © 2016 Elsevier Inc. All rights reserved.

Comparative in-vitro activity of Imipenem and Doripenem against ...

African Journals Online (AJOL)

Background: Doripenem is a recent carbapenem not commercially available in Nigeria with broad spectrum antibacterial activity against various clinical infections. Carbapenems have been shown to be the last line of agents against ESBL producing organisms. Objective: To determine the in-vitro activity of Imipenem and ...

The comet assay: assessment of in vitro and in vivo DNA damage.

Science.gov (United States)

Bajpayee, Mahima; Kumar, Ashutosh; Dhawan, Alok

2013-01-01

Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.

Understanding Mammalian Germ Line Development with In Vitro Models.

Science.gov (United States)

Martínez-Arroyo, Ana M; Míguez-Forján, Jose M; Remohí, Jose; Pellicer, Antonio; Medrano, Jose V

2015-09-15

Germ line development is crucial in organisms with sexual reproduction to complete their life cycle. In mammals, knowledge about germ line development is based mainly on the mouse model, in which genetic and epigenetic events are well described. However, little is known about how germ line development is orchestrated in humans, especially in the earliest stages. New findings derived from human in vitro models to obtain germ cells can shed light on these questions. This comprehensive review summarizes the current knowledge about mammalian germ line development, emphasizing the state of the art obtained from in vitro models for germ cell-like cell derivation. Current knowledge of the pluripotency cycle and germ cell specification has allowed different in vitro strategies to obtain germ cells with proven functionality in mouse models. Several reports during the last 10 years show that in vitro germ cell derivation with proven functionality to generate a healthy offspring is possible in mice. However, differences in the embryo development and pluripotency potential between human and mouse make it difficult to extrapolate these results. Further efforts on both human and mouse in vitro models to obtain germ cells from pluripotent stem cells may help to elucidate how human physiological events take place; therefore, therapeutic strategies can also be considered.

In vitro differentiation of rat spermatogonia into round spermatids in tissue culture.

Science.gov (United States)

Reda, A; Hou, M; Winton, T R; Chapin, R E; Söder, O; Stukenborg, J-B

2016-09-01

Do the organ culture conditions, previously defined for in vitro murine male germ cell differentiation, also result in differentiation of rat spermatogonia into post-meiotic germ cells exhibiting specific markers for haploid germ cells? We demonstrated the differentiation of rat spermatogonia into post-meiotic cells in vitro, with emphasis on exhibiting, protein markers described for round spermatids. Full spermatogenesis in vitro from immature germ cells using an organ culture technique in mice was first reported 5 years ago. However, no studies reporting the differentiation of rat spermatogonia into post-meiotic germ cells exhibiting the characteristic protein expression profile or into functional sperm have been reported. Organ culture of testicular fragments of 5 days postpartum (dpp) neonatal rats was performed for up to 52 days. Evaluation of microscopic morphology, testosterone levels, mRNA and protein expression as measured by RT-qPCR and immunostaining were conducted to monitor germ cell differentiation in vitro. Potential effects of melatonin, Glutamax® medium, retinoic acid and the presence of epidydimal fat tissue on the spermatogenic process were evaluated. A minimum of three biological replicates were performed for all experiments presented in this study. One-way ANOVA, ANOVA on ranks and student's t-test were applied to perform the statistical analysis. Male germ cells, present in testicular tissue pieces grown from 5 dpp rats, exhibited positive protein expression for Acrosin and Crem (cAMP (cyclic adenosine mono phosphate) response element modulator) after 52 days of culture in vitro. Intra-testicular testosterone production could be observed after 3 days of culture, while when epididymal fat tissue was added, spontaneous contractility of cultured seminiferous tubules could be observed after 21 days. However, no supportive effect of the supplementation with any factor or the co-culturing with epididymal fat tissue on germ cell differentiation in

Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests.

Science.gov (United States)

Cassells, Alan C

2012-01-01

The ability to establish and grow plant cell, organ, and tissue cultures has been widely exploited for basic and applied research, and for the commercial production of plants (micro-propagation). Regardless of whether the application is for research or commerce, it is essential that the cultures be established in vitro free of biological contamination and be maintained as aseptic cultures during manipulation, growth, and storage. The risks from microbial contamination are spurious experimental results due to the effects of latent contaminants or losses of valuable experimental or commercial cultures. Much of the emphasis in culture contamination management historically focussed on the elimination of phytopathogens and the maintenance of cultures free from laboratory contamination by environmental bacteria, fungi (collectively referred to as "vitro pathogens", i.e. pathogens or environmental micro-organisms which cause culture losses), and micro-arthropods ("vitro pests"). Microbial contamination of plant tissue cultures is due to the high nutrient availability in the almost universally used Murashige and Skoog (Physiol Plant 15:473-497, 1962) basal medium or variants of it. In recent years, it has been shown that many plants, especially perennials, are at least locally endophytically colonized intercellularly by bacteria. The latter, and intracellular pathogenic bacteria and viruses/viroids, may pass latently into culture and be spread horizontally and vertically in cultures. Growth of some potentially cultivable endophytes may be suppressed by the high salt and sugar content of the Murashige and Skoog basal medium and suboptimal temperatures for their growth in plant tissue growth rooms. The management of contamination in tissue culture involves three stages: disease screening (syn. disease indexing) of the stock plants with disease and endophyte elimination where detected; establishment and pathogen and contaminant screening of established initial cultures

In-vitro study on calcium carbonate crystal growth mediated by organic matrix extracted from fresh water pearls

International Nuclear Information System (INIS)

Ma Yufei; Qiao Li; Feng Qingling

2012-01-01

For the purpose of studying the mediation of organic matrix on the crystallization of calcium carbonate, water soluble matrix (WSM), acid soluble matrix (ASM) and acid insoluble matrix (AIM) were extracted from aragonite pearls and vaterite pearls respectively. Then, in-vitro calcium carbonate crystallization experiments under the control of these six organic matrices were carried out in the present study. Scanning electron microscopy (SEM) was utilized to observe the morphology of CaCO 3 and Raman spectroscopy as a powerful technique was used to distinguish the crystal polymorph. Influences of the six kinds of organic matrices on the calcium carbonate crystal growth are proposed. ASM of vaterite pearls can induce vaterite to crystallize and WSM of aragonite pearls mediates to produce aragonite crystals. The single AIM membranes of the two pearls have no pronounced effect on the CaCO 3 crystallization. Additionally, the crystal size obtained with the additive of WSM of the two kinds of pearls is smaller than that with the additive of ASM. Moreover, self-assembly phenomenon in the biomineralization process and the distorted morphology calcite are observed. Current results demonstrate important aspects of matrix protein-controlled crystallization, which is beneficial to the understanding of nacre biomineralization mechanism. Further study of the precise control of these matrix proteins on CaCO 3 crystal growth is being processed. - Highlights: ► WSM, ASM and AIM are extracted from aragonite pearls and vaterite pearls. ► ASM of vaterite pearl induces vaterite. ► WSM of aragonite pearl mediates to produce aragonite. ► WSM can fine control crystal size smaller than that with the additive of ASM. ► Self-assembly and the distorted calcite existed in the mineralization process.

Molecular ecotoxicology of nanosilver guided using in vitro prognosis

DEFF Research Database (Denmark)

Hayashi, Yuya; Heckmann, Lars-Henrik; Engelmann, Péter

2012-01-01

To study the molecular and cellular basis of silver nanoparticle (AgNP) toxicity, we here used a recently established in vitro model of earthworm coelomocytes in comparison to the conventional in vivo molecular ecotoxicology approach. Compared to the latter where the test organisms are exposed...... to NPs of interest held in an environmental matrix, in vitro models benefit from the ease of controlling exposure conditions in a defined set of biochemical milieus that NPs may encounter. The AgNPs tested in the present study originated from the same source, but to enhance the colloidal stability...... in the in vitro test media the NPs were pre-treated with serum proteins. In addition to physical characterisation of AgNPs, the active silver ion fraction was measured (in serum-supplemented cell culture medium and in soil pore-water). Using flow cytometry and atomic absorption spectrophotometry, we show...

Evaluation of air-interfaced Calu-3 cell layers for investigation of inhaled drug interactions with organic cation transporters in vitro.

Science.gov (United States)

Mukherjee, Manali; Pritchard, D I; Bosquillon, C

2012-04-15

A physiologically pertinent in vitro model is urgently needed for probing interactions between inhaled drugs and the organic cation transporters (OCT) in the bronchial epithelium. This study evaluated OCT expression, functionality, inhibition by common inhaled drugs and impact on formoterol transepithelial transport in layers of human bronchial epithelial Calu-3 cells grown at an air-liquid interface. 21 day old Calu-3 layers expressed OCT1, OCT3, OCTN1 and OCTN2 whereas OCT2 could not be detected. Quantification of the cellular uptake of the OCT substrate ASP(+) in presence of inhibitors suggested several OCT were functional at the apical side of the cell layers. ASP(+) uptake was reduced by the bronchodilators formoterol, salbutamol (albuterol), ipratropium and the glucocorticoid budesonide. However, the OCT inhibitory properties of the two β(2)-mimetics were suppressed at therapeutically relevant concentrations. The absorptive permeability of formoterol across the cell layers was enhanced at a high drug concentration shown to decrease ASP(+) uptake by ∼50% as well as in presence of the OCT inhibitor tetraethylammonium (TEA). Secretory transport was unaffected by the drug concentration but was reduced by TEA. Our data indicate air-interfaced Calu-3 layers offer a low-cost in vitro model suitable for assessing inhaled drug-OCT interactions in the bronchial epithelium. Copyright © 2011 Elsevier B.V. All rights reserved.

Sarcocystis sinensis is the most prevalent thick-walled Sarcocystis species in beef on sale for consumers in Germany.

Science.gov (United States)

Moré, G; Pantchev, A; Skuballa, J; Langenmayer, M C; Maksimov, P; Conraths, F J; Venturini, M C; Schares, G

2014-06-01

Bovines are intermediate hosts of Sarcocystis cruzi, Sarcocystis hirsuta, and Sarcocystis hominis, which use canids, felids, or primates as definitive hosts, respectively. Cattle represent also intermediate hosts of Sarcocystis sinensis, but the definitive hosts of this parasite are not yet known. Sarcocystosis in cattle is frequently asymptomatic. The infection is characterized by the presence of thin-walled (S. cruzi) or thick-walled muscle cysts or sarcocysts (S. hominis, S. sinensis, and S. hirsuta). Recent reports suggest high prevalence of the zoonotic S. hominis in beef in Europe. We therefore aimed at differentiating Sarcocystis spp. in beef offered to consumers in Germany using molecular and microscopical methods, focusing on those species producing thick-walled sarcocysts. A total of 257 beef samples were obtained from different butcheries and supermarkets in Germany and processed by conventional and multiplex real-time PCR. In addition, 130 of these samples were processed by light microscopy and in 24.6% thick-walled cysts were detected. Transmission electron microscopical analysis of six of these samples revealed an ultrastructural cyst wall pattern compatible with S. sinensis in five samples and with S. hominis in one sample. PCR-amplified 18S ribosomal DNA (rDNA) fragments of 28 individual thick-walled cysts were sequenced, and sequence identities of ≥98% with S. sinensis (n = 22), S. hominis (n = 5) and S. hirsuta (n = 1) were observed. Moreover, nine Sarcocystis sp. 18S rDNA full length gene sequences were obtained, five of S. sinensis, three of S. hominis, and one of S. hirsuta. Out of all samples (n = 257), 174 (67.7%) tested positive by conventional PCR and 179 (69.6%) by multiplex real-time PCR for Sarcocystis spp. Regarding individual species, 134 (52%), 95 (37%), 17 (6.6%), and 16 (6.2%) were positive for S. cruzi, S. sinensis, S. hirsuta, and S. hominis, respectively. In conclusion, S. sinensis is the most prevalent thick

Evaluation of the in vitro glycemic index of a fiber-rich extruded breakfast cereal produced with organic passion fruit fiber and corn flour

Directory of Open Access Journals (Sweden)

Maria Gabriela Vernaza Leoro

2010-12-01

Full Text Available The aim of this study was to determine the influence of process parameters and Passion Fruit Fiber (PFF addition on the Glycemic Index (GI of an extruded breakfast cereal. A 2³ Central Composite Rotational Design (CCRD was used, with the following independent variables: raw material moisture content (18-28%, 2nd and 3rd barrel zone temperatures (120-160 ºC, and PFF (0-30%. Raw materials (organic corn flour and organic PFF were characterized as to their proximate composition, particle size, and in vitro GI. The extrudates were characterized as to their in vitro GI. The Response Surface Methodology (RSM and Principal Component Analysis (PCA were used to analyze the results. Corn flour and PFF presented 8.55 and 7.63% protein, 2.61 and 0.60% fat, 0.52 and 6.17% ash, 78.77 and 78.86% carbohydrates (3 and 64% total dietary fiber, respectively. The corn flour particle size distribution was homogeneous, while PFF presented a heterogeneous particle size distribution. Corn flour and PFF presented values of GI of 48 and 45, respectively. When using RSM, no effect of the variables was observed in the GI of the extrudates (average value of 48.41, but PCA showed that the GI tended to be lower when processing at lower temperatures (158 ºC. When compared to white bread, the extrudates showed a reduction of the GI of up to 50%, and could be considered an interesting alternative in weight and glycemia control diets.

Recapitulation of physiological spatiotemporal signals promotes in vitro formation of phenotypically stable human articular cartilage

Science.gov (United States)

Wei, Yiyong; Zhou, Bin; Bernhard, Jonathan; Robinson, Samuel; Burapachaisri, Aonnicha; Guo, X. Edward

2017-01-01

Standard isotropic culture fails to recapitulate the spatiotemporal gradients present during native development. Cartilage grown from human mesenchymal stem cells (hMSCs) is poorly organized and unstable in vivo. We report that human cartilage with physiologic organization and in vivo stability can be grown in vitro from self-assembling hMSCs by implementing spatiotemporal regulation during induction. Self-assembling hMSCs formed cartilage discs in Transwell inserts following isotropic chondrogenic induction with transforming growth factor β to set up a dual-compartment culture. Following a switch in the basal compartment to a hypertrophic regimen with thyroxine, the cartilage discs underwent progressive deep-zone hypertrophy and mineralization. Concurrent chondrogenic induction in the apical compartment enabled the maintenance of functional and hyaline cartilage. Cartilage homeostasis, chondrocyte maturation, and terminal differentiation markers were all up-regulated versus isotropic control groups. We assessed the in vivo stability of the cartilage formed under different induction regimens. Cartilage formed under spatiotemporal regulation in vitro resisted endochondral ossification, retained the expression of cartilage markers, and remained organized following s.c. implantation in immunocompromised mice. In contrast, the isotropic control groups underwent endochondral ossification. Cartilage formed from hMSCs remained stable and organized in vivo. Spatiotemporal regulation during induction in vitro recapitulated some aspects of native cartilage development, and potentiated the maturation of self-assembling hMSCs into stable and organized cartilage resembling the native articular cartilage. PMID:28228529

Asymbiotic in vitro seed propagation of Dendrobium.

Science.gov (United States)

Teixeira da Silva, Jaime A; Tsavkelova, Elena A; Ng, Tzi Bun; Parthibhan, S; Dobránszki, Judit; Cardoso, Jean Carlos; Rao, M V; Zeng, Songjun

2015-10-01

The ability to germinate orchids from seeds in vitro presents a useful and viable method for the propagation of valuable germplasm, maintaining the genetic heterogeneity inherent in seeds. Given the ornamental and medicinal importance of many species within the genus Dendrobium, this review explores in vitro techniques for their asymbiotic seed germination. The influence of abiotic factors (such as temperature and light), methods of sterilization, composition of basal media, and supplementation with organic additives and plant growth regulators are discussed in context to achieve successful seed germination, protocorm formation, and further seedling growth and development. This review provides both a basis for the selection of optimal conditions, and a platform for the discovery of better ones, that would allow the development of new protocols and the exploration of new hypotheses for germination and conservation of Dendrobium seeds and seedlings.

Benchmarking organic micropollutants in wastewater, recycled water and drinking water with in vitro bioassays.

Science.gov (United States)

Escher, Beate I; Allinson, Mayumi; Altenburger, Rolf; Bain, Peter A; Balaguer, Patrick; Busch, Wibke; Crago, Jordan; Denslow, Nancy D; Dopp, Elke; Hilscherova, Klara; Humpage, Andrew R; Kumar, Anu; Grimaldi, Marina; Jayasinghe, B Sumith; Jarosova, Barbora; Jia, Ai; Makarov, Sergei; Maruya, Keith A; Medvedev, Alex; Mehinto, Alvine C; Mendez, Jamie E; Poulsen, Anita; Prochazka, Erik; Richard, Jessica; Schifferli, Andrea; Schlenk, Daniel; Scholz, Stefan; Shiraishi, Fujio; Snyder, Shane; Su, Guanyong; Tang, Janet Y M; van der Burg, Bart; van der Linden, Sander C; Werner, Inge; Westerheide, Sandy D; Wong, Chris K C; Yang, Min; Yeung, Bonnie H Y; Zhang, Xiaowei; Leusch, Frederic D L

2014-01-01

Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring.

In vitro fermentation of browse species using goat rumen fluid in relation to browse polyphenol content and composition

NARCIS (Netherlands)

Mengistu, G.; Karonen, M.; Salminen, J.P.; Hendriks, W.H.; Pellikaan, W.F.

2017-01-01

The effect of browse species tannins (using polyethylene glycol, PEG 6000) on in vitro gas production (GP), methane (CH 4 ), ammonia (NH 3 ), volatile fatty acids (VFA) and in vitro organic matter digestibility (IVOMD) were studied. Approximately 0.5 g of air-dried A. etbaica, C. farinosa, C.

In vitro fermentation of browse species using goat rumen fluid in relation to browse polyphenol content and composition

NARCIS (Netherlands)

Mengistu, G.; Karonen, M; Salminen, J-P; Hendriks, W. H.; Pellikaan, Wilbert F

2017-01-01

The effect of browse species tannins (using polyethylene glycol, PEG 6000) on in vitro gas production (GP), methane (CH4), ammonia (NH3), volatile fatty acids (VFA) and in vitro organic matter digestibility (IVOMD) were studied. Approximately 0.5 g of air-dried A. etbaica, C. farinosa, C. tomentosa,

In vitro activity of certain drugs in combination with plant extracts ...

African Journals Online (AJOL)

STORAGESEVER

2009-09-01

Sep 1, 2009 ... organisms (Hugo et al., 1993; Levinson and Jawetz,. 2002). Drug synergism between known antimicrobial agents and bioactive plant extracts is .... Ibezim EC, Esimone CO, Nnamani PO, Onyishi IV, Brown SA, Obodo. CE (2006). In vitro study of the interaction between some fluoroquinolones and extracts of ...

The effect of some organic compounds and NAA application on the in vitro growth of the black orchid (Coelogyne pandurata Lindl.

Directory of Open Access Journals (Sweden)

RINI UNTARI

2006-10-01

Full Text Available Coelogyne pandurata Lindl. is a lowland epiphytic orchid that has pale green flowers with typically black markings on the lips. This species conventionally propagated asexually by rhizome. This orchid is now facing a great conservation problem and threatened to extinction due to human exploitation. That is why conservation activities such as developing proper cultivation, are urgently required. An in vitro experiment was conducted at the Laboratory of Tissue Culture at Bogor Botanic Garden. The experiment was carried out using a completely randomized design with two factors and ten replications. The treatments were the supplementation of the basal media i.e. Vacin and Went added with sugar, activated charcoal and agar, with 30 different combinations of organic compounds i.e., coconut water 250 mL/L, banana 150 g/L, potato 200 g/L, sweet potato 150 g/L, soybean 150 g/L and no organic compound as a control, and application of NAA (0, 5, 10, 15, and 20 ppm. The result showed that there was a significant effect of the organic compounds and NAA application on the length and the number of roots, height, number of leaves and number of shoots produced by the explants. The combination of sweet potato 150 g/L without NAA application showed the best result.

Assembly of cells and vesicles for organ engineering

International Nuclear Information System (INIS)

Taguchi, Tetsushi

2011-01-01

The development of materials and technologies for the assembly of cells and/or vesicles is a key for the next generation of tissue engineering. Since the introduction of the tissue engineering concept in 1993, various types of scaffolds have been developed for the regeneration of connective tissues in vitro and in vivo. Cartilage, bone and skin have been successfully regenerated in vitro, and these regenerated tissues have been applied clinically. However, organs such as the liver and pancreas constitute numerous cell types, contain small amounts of extracellular matrix, and are highly vascularized. Therefore, organ engineering will require the assembly of cells and/or vesicles. In particular, adhesion between cells/vesicles will be required for regeneration of organs in vitro. This review introduces and discusses the key technologies and materials for the assembly of cells/vesicles for organ regeneration. (topical review)

In vivo and in vitro studies of cartilage differentiation in altered gravities

Science.gov (United States)

Montufar-Solis, D.; Duke, P. J.; D'Aunno, D.

The in vivo model our laboratory uses for studies of cartilage differentiation in space is the rat growth plate. Differences between missions, and in rat age and recovery times, provided differing results from each mission. However, in all missions, proliferation and differentiation of chondrocytes in the epiphyseal plate of spaceflown rats was altered as was matrix organization. In vitro systems, necessary complements to in vivo work, provide some advantages over the in vivo situation. In vitro, centrifugation of embryonic limb buds suppressed morphogenesis due to precocious differentiation, and changes in the developmental pattern suggest the involvement of Hox genes. In space, embryonic mouse limb mesenchyme cells differentiating in vitro on IML-1 had smoother membranes and lacked matrix seen in controls. Unusual formations, possibly highly ruffled membranes, were found in flight cultures. These results, coupled with in vivo centrifugation studies, show that in vivo or in vitro, the response of chondrocytes to gravitational changes follows Hert's curve as modified by Simon, i.e. decreased loading decreases differentiation, and increased loading speeds it up, but only to a point. After that, additional increases again slow down chondrogenesis.

Osteoblast Differentiation and Bone Matrix Formation In Vivo and In Vitro.

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Blair, Harry C; Larrouture, Quitterie C; Li, Yanan; Lin, Hang; Beer-Stoltz, Donna; Liu, Li; Tuan, Rocky S; Robinson, Lisa J; Schlesinger, Paul H; Nelson, Deborah J

2017-06-01

We review the characteristics of osteoblast differentiation and bone matrix synthesis. Bone in air breathing vertebrates is a specialized tissue that developmentally replaces simpler solid tissues, usually cartilage. Bone is a living organ bounded by a layer of osteoblasts that, because of transport and compartmentalization requirements, produce bone matrix exclusively as an organized tight epithelium. With matrix growth, osteoblasts are reorganized and incorporated into the matrix as living cells, osteocytes, which communicate with each other and surface epithelium by cell processes within canaliculi in the matrix. The osteoblasts secrete the organic matrix, which are dense collagen layers that alternate parallel and orthogonal to the axis of stress loading. Into this matrix is deposited extremely dense hydroxyapatite-based mineral driven by both active and passive transport and pH control. As the matrix matures, hydroxyapatite microcrystals are organized into a sophisticated composite in the collagen layer by nucleation in the protein lattice. Recent studies on differentiating osteoblast precursors revealed a sophisticated proton export network driving mineralization, a gene expression program organized with the compartmentalization of the osteoblast epithelium that produces the mature bone matrix composite, despite varying serum calcium and phosphate. Key issues not well defined include how new osteoblasts are incorporated in the epithelial layer, replacing those incorporated in the accumulating matrix. Development of bone in vitro is the subject of numerous projects using various matrices and mesenchymal stem cell-derived preparations in bioreactors. These preparations reflect the structure of bone to variable extents, and include cells at many different stages of differentiation. Major challenges are production of bone matrix approaching the in vivo density and support for trabecular bone formation. In vitro differentiation is limited by the organization and

Presentation of an experimental method to induce in vitro ("organ chambers") respiratory acidosis and its effect on vascular reactivity.

Science.gov (United States)

Nadai, Tales Rubens de; Silveira, Ana Paula Cassiano; Monteiro, Ariadne Santana e Neves; Campos, Debora Ribeiro; Carvalho, Marco Tulio Rezende de; Albuquerque, Agnes Afrodite Sumarelli; Celotto, Andrea Carla; Evora, Paulo Roberto Barbosa

2014-11-01

To create in vitro a model to generate acidosis by CO2 bubbling "organ chambers", which would be useful for researchers that aim to study the effects of acid-base disturbs on the endothelium-dependent vascular reactivity. Eighteen male Wistar rats (230-280 g) were housed, before the experiments, under standard laboratory conditions (12h light/dark cycle at 21°C), with free access to food and water. The protocol for promoting in vitro respiratory acidosis was carried out by bubbling increased concentrations of CO2. The target was to achieve an ideal way to decrease the pH gradually to a value of approximately 6.6.It was used, initially, a gas blender varying concentrations of the carbogenic mixture (95% O2 + 5% CO2) and pure CO2. 1) 100% CO2, pH variation very fast, pH minimum 6.0; 2) 90%CO2 pH variation bit slower, pH minimum 6.31; 3) 70%CO2, pH variation slower, pH minimum 6.32; 4) 50% CO2, pH variation slower, pH minimum 6:42; 5) 40 %CO2, Adequate record, pH minimum 6.61, and; 6) 30 %CO2 could not reach values below pH minimum 7.03. Based on these data the gas mixture (O2 60% + CO2 40%) was adopted. This gas mixture (O2 60% + CO2 40%) was effective in inducing respiratory acidosis at a speed that made, possible the recording of isometric force.

The combined effects of vinclozolin and procymidone do not deviate from expected additivity in vitro and in vivo

DEFF Research Database (Denmark)

Nellemann, Christine Lydia; Dalgaard, Majken; Lam, Henrik Rye

2003-01-01

The combination effects of the well-known antiandrogenic fungicides, vinclozolin and procymidone, were tested both in vitro and in vivo. In vitro both vinclozolin and procymidone significantly inhibited the binding of agonist to the androgen receptor with the concentration that resulted in 50......% inhibition (IC50) values of 0.1 and 0.6 muM, respectively. By applying the isobole method, the effect of combining the two pesticides in vitro was found to be additive. In castrated testosterone-treated rats the administration of vinclozolin starting at 10 mg/kg led to a decrease in organ weight of all...... tested reproductive organs. The levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were increased significantly with doses of 100 mg/kg vinclozolin and above. Expression of the androgen-responsive gene, TRPM-2, was increased starting at 100 mg/kg vinclozolin. For procymidone...

The combined effects of vinclozolin and procymidone do not deviate from expected additivity in vitro and in vivo.

Science.gov (United States)

Nellemann, Christine; Dalgaard, Majken; Lam, Henrik Rye; Vinggaard, Anne Marie

2003-02-01

The combination effects of the well-known antiandrogenic fungicides, vinclozolin and procymidone, were tested both in vitro and in vivo. In vitro both vinclozolin and procymidone significantly inhibited the binding of agonist to the androgen receptor with the concentration that resulted in 50% inhibition (IC(50)) values of 0.1 and 0.6 micro M, respectively. By applying the isobole method, the effect of combining the two pesticides in vitro was found to be additive. In castrated testosterone-treated rats the administration of vinclozolin starting at 10 mg/kg led to a decrease in organ weight of all tested reproductive organs. The levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were increased significantly with doses of 100 mg/kg vinclozolin and above. Expression of the androgen-responsive gene, TRPM-2, was increased starting at 100 mg/kg vinclozolin. For procymidone, reproductive organ weights were diminished at 10 mg/kg and LH was increased at a concentration of 25 mg/kg and above, compared to the testosterone-treated controls. FSH was significantly increased only at 25 mg/kg procymidone. The studied gene expressions were changed by 100 mg/kg procymidone. Dosing the animals with a combination of a 1:1 mixture of vinclozolin and procymidone resulted in a weight reduction in the reproductive organs and an increase of serum LH and FSH as early as with 10 mg/kg combined dose. The relative expressions of TRPM-2 and PBP C3 were changed compared to controls at 100 mg/kg. The level of 5-HT in the rat brain was increased after a dose of 10 mg/kg. Using the isobole method, comparisons of the observed and predicted effects assuming additivity on reproductive organ weights, hormone levels, and gene expression showed agreement and thus the combination effects are suggested to be additive in vivo as well as in vitro.

Phytochemical And In Vitro Antimicrobial Assay Of The Leaf Extract ...

African Journals Online (AJOL)

This study has justified the traditional use of this plant for the treatment of stomach discomfort, diarrhea, dysentery and as a remedy for wound healing whose causative agents are some of the organisms used in this study. Keywords: Antimicrobial, Leaf Extracts,In Vitro, Phytochemical, Newbouldia laevis. African Journal of ...

In vitro pancreas organogenesis from dispersed mouse embryonic progenitors

DEFF Research Database (Denmark)

Greggio, Chiara; De Franceschi, Filippo; Figueiredo-Larsen, Evan Manuel

2014-01-01

The pancreas is an essential organ that regulates glucose homeostasis and secretes digestive enzymes. Research on pancreas embryogenesis has led to the development of protocols to produce pancreatic cells from stem cells (1). The whole embryonic organ can be cultured at multiple stages...... expanding progenitors and differentiate into endocrine, acinar and ductal cells and which spontaneously self-organize to resemble the embryonic pancreas. We show here that the in vitro process recapitulates many aspects of natural pancreas development. This culture system is suitable to investigate how...... cells cooperate to form an organ by reducing its initial complexity to few progenitors. It is a model that reproduces the 3D architecture of the pancreas and that is therefore useful to study morphogenesis, including polarization of epithelial structures and branching. It is also appropriate to assess...

Metagenomic Sequencing of an In Vitro-Simulated Microbial Community

Energy Technology Data Exchange (ETDEWEB)

Morgan, Jenna L.; Darling, Aaron E.; Eisen, Jonathan A.

2009-12-01

Background: Microbial life dominates the earth, but many species are difficult or even impossible to study under laboratory conditions. Sequencing DNA directly from the environment, a technique commonly referred to as metagenomics, is an important tool for cataloging microbial life. This culture-independent approach involves collecting samples that include microbes in them, extracting DNA from the samples, and sequencing the DNA. A sample may contain many different microorganisms, macroorganisms, and even free-floating environmental DNA. A fundamental challenge in metagenomics has been estimating the abundance of organisms in a sample based on the frequency with which the organism's DNA was observed in reads generated via DNA sequencing. Methodology/Principal Findings: We created mixtures of ten microbial species for which genome sequences are known. Each mixture contained an equal number of cells of each species. We then extracted DNA from the mixtures, sequenced the DNA, and measured the frequency with which genomic regions from each organism was observed in the sequenced DNA. We found that the observed frequency of reads mapping to each organism did not reflect the equal numbers of cells that were known to be included in each mixture. The relative organism abundances varied significantly depending on the DNA extraction and sequencing protocol utilized. Conclusions/Significance: We describe a new data resource for measuring the accuracy of metagenomic binning methods, created by in vitro-simulation of a metagenomic community. Our in vitro simulation can be used to complement previous in silico benchmark studies. In constructing a synthetic community and sequencing its metagenome, we encountered several sources of observation bias that likely affect most metagenomic experiments to date and present challenges for comparative metagenomic studies. DNA preparation methods have a particularly profound effect in our study, implying that samples prepared with

In vitro oxidation of mercury by the blood

International Nuclear Information System (INIS)

Hursh, J.B.; Sichak, S.P.; Clarkson, T.W.

1988-01-01

A method is described for studying the in vitro oxidation of mercury vapour by red blood cells at short times and with diminishing mercury vapour concentrations. It is found that for 40% red blood cell suspensions and 37 deg. C at concentrations greater than about 6 ng mercury vapour/ml, the oxidation rate is zero order, and that at lower concentrations the rate changes to first order. The effect of temperature and of added hydrogen peroxide de are studied. Results a considered in terms of the generally accepted belief that the catalase-compound I system is the main path of oxidation. If the results obtained in vitro in these experiments apply in vivo to man, it follows that inhaled mercury is carried in the blood to the brain and organs primarily as dissolved vapour rather than as inorganic mercury ions. (author)

Chloroform and trichloroethylene uptake from water into human skin in vitro: Kinetics and risk implications

International Nuclear Information System (INIS)

Bogen, K.T.; Keating, G.A.; Vogel, J.S.

1995-03-01

A model recently proposed by the US Environmental Protection Agency (EPA) predicts that short-term dermal uptakes of organic environmental water contaminants are proportional to the square root of exposure time. The model appears to underestimate dermal uptake, based on very limited in vivo uptake data obtained primarily using human subjects. To further assess this model, we examined in vitro dermal uptake kinetics for aqueous organic chemicals using accelerator mass spectrometry (AMS). Specifically, we examined the kinetics of in vitro dermal uptake of 14 C-labeled chloroform and trichloroethylene from dilute (5-ppb) aqueous solutions using full-thickness human cadaver skin exposed for (≤1 hr)

Microengineered physiological biomimicry: organs-on-chips.

Science.gov (United States)

Huh, Dongeun; Torisawa, Yu-suke; Hamilton, Geraldine A; Kim, Hyun Jung; Ingber, Donald E

2012-06-21

Microscale engineering technologies provide unprecedented opportunities to create cell culture microenvironments that go beyond current three-dimensional in vitro models by recapitulating the critical tissue-tissue interfaces, spatiotemporal chemical gradients, and dynamic mechanical microenvironments of living organs. Here we review recent advances in this field made over the past two years that are focused on the development of 'Organs-on-Chips' in which living cells are cultured within microfluidic devices that have been microengineered to reconstitute tissue arrangements observed in living organs in order to study physiology in an organ-specific context and to develop specialized in vitro disease models. We discuss the potential of organs-on-chips as alternatives to conventional cell culture models and animal testing for pharmaceutical and toxicology applications. We also explore challenges that lie ahead if this field is to fulfil its promise to transform the future of drug development and chemical safety testing.

Growing Arabidopsis in vitro: cell suspensions, in vitro culture, and regeneration.

Science.gov (United States)

Barkla, Bronwyn J; Vera-Estrella, Rosario; Pantoja, Omar

2014-01-01

An understanding of basic methods in Arabidopsis tissue culture is beneficial for any laboratory working on this model plant. Tissue culture refers to the aseptic growth of cells, organs, or plants in a controlled environment, in which physical, nutrient, and hormonal conditions can all be easily manipulated and monitored. The methodology facilitates the production of a large number of plants that are genetically identical over a relatively short growth period. Techniques, including callus production, cell suspension cultures, and plant regeneration, are all indispensable tools for the study of cellular biochemical and molecular processes. Plant regeneration is a key technology for successful stable plant transformation, while cell suspension cultures can be exploited for metabolite profiling and mining. In this chapter we report methods for the successful and highly efficient in vitro regeneration of plants and production of stable cell suspension lines from leaf explants of both Arabidopsis thaliana and Arabidopsis halleri.

Comparative analysis of perturbed molecular pathways identified in in vitro and in vivo toxicology studies

NARCIS (Netherlands)

Wiesinger, Martin; Mayer, Bernd; Jennings, Paul; Lukas, Arno

The development of in vitro toxicological testing strategies are hampered by the difficulty in extrapolation to the intact organism. Academic toxicological literature contains a wealth of mechanistically rich information, especially arising from omic studies, which could potentially be utilized to

Chemical Carcinogen (Hydrazine et al.) Induced Carcinogenesis of Human Diploid Cells in Vitro

Science.gov (United States)

1982-09-07

untreated cell popula- pared from the stock solutions in complete growth tions were seeded onto CES in vitro. The CES medium (CIM). Thee solutions were added...time from induction pared to metastases IFig. IBl. We want to imply to neoplasia of 6 to 10 wk instead of 1 to 1.5 yr. that these chemical carcinogen...This definition was used because the pathologist was ds:b celular invasion Into CE,, an organ culture, In vitro. Thw two Interpretations am not

The contribution of oxidative stress to drug-induced organ toxicity and its detection in vitro and in vivo.

Science.gov (United States)

Pereira, Claudia V; Nadanaciva, Sashi; Oliveira, Paulo J; Will, Yvonne

2012-02-01

Nowadays the 'redox hypothesis' is based on the fact that thiol/disulfide couples such as glutathione (GSH/GSSG), cysteine (Cys/CySS) and thioredoxin ((Trx-(SH)2/Trx-SS)) are functionally organized in redox circuits controlled by glutathione pools, thioredoxins and other control nodes, and they are not in equilibrium relative to each other. Although ROS can be important intermediates of cellular signaling pathways, disturbances in the normal cellular redox can result in widespread damage to several cell components. Moreover, oxidative stress has been linked to a variety of age-related diseases. In recent years, oxidative stress has also been identified to contribute to drug-induced liver, heart, renal and brain toxicity. This review provides an overview of current in vitro and in vivo methods that can be deployed throughout the drug discovery process. In addition, animal models and noninvasive biomarkers are described. Reducing post-market drug withdrawals is essential for all pharmaceutical companies in a time of increased patient welfare and tight budgets. Predictive screens positioned early in the drug discovery process will help to reduce such liabilities. Although new and more efficient assays and models are being developed, the hunt for biomarkers and noninvasive techniques is still in progress.

The chemical composition and in vitro dry matter digestibility of ...

African Journals Online (AJOL)

Leendert Snynan

Crop residues in the summer rainfall area of South Africa fulfill a strategic role in the fodder flow program ... This value is high when compared with in vitro organic matter digestibility values reported for wheat straw. (38.9%) ... Sunflower-cob residues seem to be a forage with a high energy value that might be suitable for use.

Genetic variants of genital mycoplasmas and their relation to the clinical course of inflammatory diseases of the urogenital system in women

Directory of Open Access Journals (Sweden)

M. R. Rakhmatulina

2014-01-01

Full Text Available Goal of the study. To study genetic variability of M. hominis (based on the vaa gene and M. genitalium (based on the mg192 gene derived from biological material samples taken from women with different clinical manifestations of inflammatory diseases of the urogenital system and clinically healthy women (for M. hominis. Materials and methods. Genetic variability of twenty M. hominis samples taken from patients with inflammatory diseases of the urogenital system and twenty M. hominis samples taken from patients without clinical and laboratory signs of inflammatory processes of the urogenital system was studied by the sequence analysis method. Genetic variability of eight M. genitalium samples taken from patients with different clinical signs of inflammatory diseases of the urogenital system was also examined by the sequence analysis method. Results. Three genetic variants of M. hominis based on the vaa gene were found; variant I was revealed more often in patients with clinical manifestations of inflammatory diseases (65.0%, and variant II - in clinically healthy women (60.0%. Three phylogenetic M. gentium groups based on the mg192 gene were revealed; two samples were referred to Groups I and II, and four samples were referred to Group III. Samples from each phylogenetic groups were taken from patients with clinical and laboratory manifestations of cervicitis and urethritis. Conclusion. The study data demonstrate a potential effect of the VAA surface protein on the virulence of opportunistic M. hominis pathogens.

In vitro study of proteins surface activity by tritium probe

International Nuclear Information System (INIS)

Chernysheva, M.G.; Badun, G.A.

2010-01-01

A new technique for in vitro studies of biomacromolecules interactions, their adsorption at aqueous/organic liquid interfaces and distribution in the bulk of liquid/liquid systems was developed. The method includes (1) tritium labeling of biomolecules by tritium thermal activation method and (2) scintillation phase step with organic phase, which can be concerned as a model of cellular membrane. Two globular proteins lysozyme and human serum albumin tested. We have determined the conditions of tritium labeling when labeled by-products can be easy separated by means of dialysis and size-exclusion chromatography. Scintillation phase experiments were conducted for three types of organic liquids. Thus, the influences of the nature of organic phase on proteins adsorption and its distribution in the bulk of aqueous/organic liquid system were determined. It was found that proteins possess high surface activity at aqueous/organic liquid interface. Furthermore, values of hydrophobicity of globular proteins were found by the experiment. (author)

Mechanistic analysis of solute transport in an in vitro physiological two-phase dissolution apparatus.

Science.gov (United States)

Mudie, Deanna M; Shi, Yi; Ping, Haili; Gao, Ping; Amidon, Gordon L; Amidon, Gregory E

2012-10-01

In vitro dissolution methodologies that adequately capture the oral bioperformance of solid dosage forms are critical tools needed to aid formulation development. Such methodologies must encompass important physiological parameters and be designed with drug properties in mind. Two-phase dissolution apparatuses, which contain an aqueous phase in which the drug dissolves (representing the dissolution/solubility component) and an organic phase into which the drug partitions (representing the absorption component), have the potential to provide meaningful predictions of in vivo oral bioperformance for some BCS II, and possibly some BCS IV drug products. Before such an apparatus can be evaluated properly, it is important to understand the kinetics of drug substance partitioning from the aqueous to the organic medium. A mass transport analysis was performed of the kinetics of partitioning of drug substance solutions from the aqueous to the organic phase of a two-phase dissolution apparatus. Major assumptions include pseudo-steady-state conditions, a dilute aqueous solution and diffusion-controlled transport. Input parameters can be measured or estimated a priori. This paper presents the theory and derivation of our analysis, compares it with a recent kinetic approach, and demonstrates its effectiveness in predicting in vitro partitioning profiles of three BCS II weak acids in four different in vitro two-phase dissolution apparatuses. Very importantly, the paper discusses how a two-phase apparatus can be scaled to reflect in vivo absorption kinetics and for which drug substances the two-phase dissolution systems may be appropriate tools for measuring oral bioperformance. Copyright © 2012 John Wiley & Sons, Ltd.

Towards a realistic risk characterization of complex mixtures using in vitro bioassays

NARCIS (Netherlands)

Montano Garces, M.

2013-01-01

This thesis aims to better understand and further improve the relevance and reliabilityof in vitro bioassaysfor a biobased risk characterisation of complex mixtures, with special focus on persistent organic pollutants (POPs) in sediments.

In Chapter 1 the importance of complex

In vitro assessment of environmental stress of persistent organic pollutants on the Indo-Pacific humpback dolphin.

Science.gov (United States)

Jia, Kuntong; Ding, Liang; Zhang, Lingli; Zhang, Mei; Yi, Meisheng; Wu, Yuping

2015-12-25

Persistent organic pollutants (POPs) are detected ubiquitously and are linked to range of adverse health effects. The Indo-Pacific humpback dolphin inhabited the Pearl River Estuary (PRE), China, where high concentrations of POPs have been reported. This study evaluated the threats posed by POPs in the environment to the dolphin using an in vitro system. We selected BNF(β-naphthoflavone) and four POPs (DDTs (dichlorodiphenyltrichloroethanes), CHLs(chlorides), HCHs(hexachlorocyclohexanes) and HCB(hexachlorobenzene)) which had been accumulated in the dolphin with high concentrations to treat the cultured skin fibroblast cells (ScSF cells) of the dolphin, and investigated the expression patterns of the ecological stress biomarkers CYP1A1, AHR and HSP70 in the cell line. The results showed that CYP1A1 was up-regulated after being exposed to different concentrations of BNF, DDTs and HCHs. CHLs, HCHs and HCB promoted AHR expression. HSP70 expression was increased by high concentrations of BNF and DDTs. Moreover, comet assay experiments revealed that DDTs produced higher degree of DNA damage to ScSF cells than other POPs, implying that the Indo-Pacific humpback dolphin in the PRE has been threatened by POPs accumulated in the body, especially by DDTs. Our results provided important information to assess the risk of the Indo-Pacific humpback dolphin raised by environmental POPs in vivo. Copyright © 2015 Elsevier B.V. All rights reserved.

In Vitro Procedures with Radioisotopes in Medicine. Proceedings of the Symposium on In Vitro Procedures with Radioistopes in Clinical Medicine and Research

International Nuclear Information System (INIS)

1970-01-01

Radioactive tracer techniques are now being widely used in the study of substances of medical interest in vitro. The introduction of tracer techniques often leads to a considerable improvement in established medical laboratory procedures. Tracer techniques also form the basis of certain special procedures for the measurement of hormones, vitamins and other substances in specimens of blood, urine, body tissues, etc.; these techniques are of such elegance and simplicity that they may readily be carried out in the average medical radioisotope laboratory without the need for additional specialized equipment. In none of these procedures is radioactive material administered to the patient; indeed, they do not require his direct participation at all and may well be carried out in some central laboratory on specimens sent in from a wide area. They may thus freely be used in investigations on all classes of patient and are. particularly suitable for use in regions where local facilities are limited or where field studies are to be undertaken. Progress in these applications of tracer techniques was discussed in the Symposium on In Vitro Procedures with Radioisotopes in Clinical Medicine and Research, organized by the International Atomic Energy Agency in co-operation with the World Health Organization. The symposium was held in Vienna from 8 to 12 September 1969, and the full Proceedings appear in the present volume. A total of 191 participants nominated by 32 countries and 3 international organizations attended, and the 50 papers presented, which cover the applications of tracer, techniques in cytological studies, immunological studies, derivative, dilution and enzymatic analysis, saturation analysis, thyroid function studies and radioimmunoassay, indicate the great variety of in vitro procedures now in use. Invited review papers deal with the general aspects of the various main groups of applications. Many of the procedures discussed are still in the developmental stage

3D Bioprinting of Tissue/Organ Models.

Science.gov (United States)

Pati, Falguni; Gantelius, Jesper; Svahn, Helene Andersson

2016-04-04

In vitro tissue/organ models are useful platforms that can facilitate systematic, repetitive, and quantitative investigations of drugs/chemicals. The primary objective when developing tissue/organ models is to reproduce physiologically relevant functions that typically require complex culture systems. Bioprinting offers exciting prospects for constructing 3D tissue/organ models, as it enables the reproducible, automated production of complex living tissues. Bioprinted tissues/organs may prove useful for screening novel compounds or predicting toxicity, as the spatial and chemical complexity inherent to native tissues/organs can be recreated. In this Review, we highlight the importance of developing 3D in vitro tissue/organ models by 3D bioprinting techniques, characterization of these models for evaluating their resemblance to native tissue, and their application in the prioritization of lead candidates, toxicity testing, and as disease/tumor models. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The efficacy of a nested PCR in detecting cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei var. Hominis for diagnosing scabies.

Science.gov (United States)

Hahm, J E; Kim, C W; Kim, S S

2018-04-06

A widespread scabies infestation, associated to long-term residence in nursing homes, is becoming a serious issue in developed countries. Mineral oil examination is regarded as the gold standard in diagnosing scabies, but the sensitivity of this method is generally low-approximately 50%. Molecular tests may contribute to enhance the sensitivity of current tests for laboratory diagnosis of human scabies. In this study, we developed new primers for a nested PCR for the cytochrome c oxidase subunit 1 (cox1) gene of Sarcoptes scabiei var. hominis to increase the sensitivity of a previously developed conventional PCR. Clinically suspected scabies patients underwent dermoscopy-guided skin scraping with microscopic examination. The diagnosis was positive for scabies when mites or eggs were found under the microscope, and patients were then designated as 'microscopy-positive'. Patients in the 'microscopy-negative' group presented with negative microscopic results. Skin scrapings were collected from both groups for PCR. Of the total 63 samples, 28 were microscopy-positive and 35 were negative with no differences in sex and age between the two groups. All microscopically proven scabies cases were positive with the cox1 nested PCR. Among microscopy-negative ones, S. scabiei DNA was detected in 9 samples. If sensitivity of the cox1 nested PCR is considered 100% (95% CI, 90.51-100), then sensitivity of microscopy is 75.68% (95% CI, 58.80-88.23; P = 0.004). Nested PCR can be successfully used as an alternative method for diagnosing suspected scabies patient. Therefore, infection control measures and treatments can be initiated before significant transmission occurs, minimizing the risk of outbreaks. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

The genome of the obligate intracellular parasite Trachipleistophora hominis: new insights into microsporidian genome dynamics and reductive evolution.

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Eva Heinz

Full Text Available The dynamics of reductive genome evolution for eukaryotes living inside other eukaryotic cells are poorly understood compared to well-studied model systems involving obligate intracellular bacteria. Here we present 8.5 Mb of sequence from the genome of the microsporidian Trachipleistophora hominis, isolated from an HIV/AIDS patient, which is an outgroup to the smaller compacted-genome species that primarily inform ideas of evolutionary mode for these enormously successful obligate intracellular parasites. Our data provide detailed information on the gene content, genome architecture and intergenic regions of a larger microsporidian genome, while comparative analyses allowed us to infer genomic features and metabolism of the common ancestor of the species investigated. Gene length reduction and massive loss of metabolic capacity in the common ancestor was accompanied by the evolution of novel microsporidian-specific protein families, whose conservation among microsporidians, against a background of reductive evolution, suggests they may have important functions in their parasitic lifestyle. The ancestor had already lost many metabolic pathways but retained glycolysis and the pentose phosphate pathway to provide cytosolic ATP and reduced coenzymes, and it had a minimal mitochondrion (mitosome making Fe-S clusters but not ATP. It possessed bacterial-like nucleotide transport proteins as a key innovation for stealing host-generated ATP, the machinery for RNAi, key elements of the early secretory pathway, canonical eukaryotic as well as microsporidian-specific regulatory elements, a diversity of repetitive and transposable elements, and relatively low average gene density. Microsporidian genome evolution thus appears to have proceeded in at least two major steps: an ancestral remodelling of the proteome upon transition to intracellular parasitism that involved reduction but also selective expansion, followed by a secondary compaction of genome

Pituitary tumour causing gigantism. Morphology and in vitro hormone secretion.

Science.gov (United States)

Anniko, M; Ritzén, E M

1986-01-01

True gigantism with overproduction of growth hormone (GH) and prolactin (PRL) was diagnosed in a 13-year-old boy. The clinical history indicated that the tumour had caused an oversecretion of GH since the age of 4-5 years. At diagnosis, the sella turcica was markedly enlarged. No infiltrative growth was noted at surgery. Endocrine investigations showed elevated GH and PRL secretion. Light and electron microscopy of tumour tissue revealed densely packed pleomorphic cells of both GH and PRL type. In addition, oncocyte-like cells were observed. Organ culture of pieces of tumour tissue demonstrated continued secretion of GH and PRL into the medium for more than 5 days in vitro. Addition of bromocriptine to the medium caused a rapid decline in PRL secretion while GH secretion remained the same. X-ray irradiation in vitro also caused a decrease in PRL secretion. These effects of bromocriptine and X-ray on hormone secretion in vitro mirrored the corresponding effect of treatment, when the patient showed signs of tumour recurrence after pituitary surgery. It is concluded that also in childhood, the in vitro response of tumour tissue to various treatments may be explored as a possible way to predict the efficacy of pharmacological or irradiation treatment of pituitary tumours.

Peri-Implant Strain in an In Vitro Model.

Science.gov (United States)

Hussaini, Souheil; Vaidyanathan, Tritala K; Wadkar, Abhinav P; Quran, Firas A Al; Ehrenberg, David; Weiner, Saul

2015-10-01

An in vitro experimental model was designed and tested to determine the influence that peri-implant strain may have on the overall crestal bone. Strain gages were attached to polymethylmethacrylate (PMMA) models containing a screw-type root form implant at sites 1 mm from the resin-implant interface. Three different types of crown superstructures (cemented, 1-screw [UCLA] and 2-screw abutment types) were tested. Loading (1 Hz, 200 N load) was performed using a MTS Mechanical Test System. The strain gage data were stored and organized in a computer for statistical treatment. Strains for all abutment types did not exceed the physiological range for modeling and remodeling of cancellous bone, 200-2500 με (microstrain). For approximately one-quarter of the trials, the strain values were less than 200 με the zone for bone atrophy. The mean microstrain obtained was 517.7 με. In conclusion, the peri-implant strain in this in vitro model did not exceed the physiologic range of bone remodeling under axial occlusal loading.

What is «to act for an end»? Francisco Suarez’ answer in the ms De beatitudine (1579 and in De ultimo fine hominis (1628

Directory of Open Access Journals (Sweden)

Rosa Colmenarejo Fernández

2018-05-01

Full Text Available The influence of the work and teaching of Francisco Suárez (Granada, 1548-Lisbon, 1617 in the shaping of modern thought has been widely studied in areas such as political philosophy and law, metaphysics, and epistemology. However, the moral philosophy of Suarez, in particular that which, as he himself explains, was needed as a basis for his moral theology, has yet to be analyzed. In this paper, we propose to elucidate some aspects of Suarez’s theory of human agency final causality. We compare two of his commentaries, from different periods, on the I-IIae of Thomas Aquinas’ Summa theologiae: the unpublished manuscript De beatitudine (1579 and Suarez’s treatise De ultimo fine hominis, the first of five treatises on morality posthumously published by Baltasar Álvares (1628. The question analyzed here, the nature of final causality, is assumed as a heuristic instrument to acquaint readers with both the sources studied and the philosophical import of Suárez’ doctrine on morality. The findings suggest the following: i the closeness, still with doubts, of Suárez’ 1579 commentary to Thomas’ doctrine of human agency final causality; (ii in the 1628 commentary, his partial assumption of Thomas’ doctrine on the role of intellect in free choice, and his clear determination in the assertion that the locus finis is the will. Working with these nuances, we illustrate some connection points that show both how 16th century scholasticism deals with philosophical tradition to explain a new image of man, and how subsequent moral philosophy is influenced by these theories while rejecting their theological foundation.

Organic Additives Improves the in Vitro Growth of Native Orchid Vanda helvola Blume

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Devina DAVID

2015-06-01

Full Text Available In vitro seed germination has been proven to be the most efficient technique to propagate orchid. The application of this aseptic technique has contributed to conservation of many endangered orchid species. In this study, undehisced capsules of Vanda helvola Blume were collected from Orchid Conservation Centre in Lagud Sebrang Agriculture Park, after 120 days from hand pollination and aseptically cultured on three types of basal media such as Murashige and Skoog (MS, Knudson C (KC and Vacin and Went (VW. After 90 days of culture, 66.40 ± 4.14% of seeds successfully germinated on KC medium. The effect of organic additives such as tomato juice, coconut water, peptone and yeast extract at different level of concentrations in KC basal medium were also tested on seed germination and seedling development of this native orchid. After 90 days of culture, over 90% of seeds were tremendously germinated on KC medium supplemented with 10% or 15% (v/v of tomato juice. The incorporation of peptone at 0.1% (w/v in KC basal media promoted rapid development of protocorm to seedling. Seedlings on this treatment produced an average of three leaves and two roots after 90 days of culture and were successfully acclimatized.

Organ culture of C57BL/6 mouse arteries with LPS as an in vitro model of vascular inflammation

DEFF Research Database (Denmark)

Outzen, Emilie Middelbo; Mehryar, Rahila; Boonen, Harrie C.M.

Background: Vascular inflammation is believed to be involved in the pathogenesis of various cardiovascular diseases, the study of which often involves use of the mouse strain C57BL/6. In vivo studies can, however, be difficult to control and interpret. Aim of the study: To set up and characterise...... an in vitro model for studying vascular inflammation and function in cultured arteries from C57BL/6 mice. Methods: Segments of abdominal aorta and mesenteric arteries (MA) were incubated for 24 hours at 37̊C and 95% O2/5% CO2 in DMEM ± 100 ng/mL LPS. Aorta segments were frozen for molecular studies...... was achieved at a normalisation factor of 0.9 (0.91 ± 0.06, mean ± SEM, n = 9) as observed (0.85 ± 0.06, mean ± SEM, n = 3) and previously described in rat MA (Mulvany and Halpern, 1977). Furthermore, preliminary findings show that organ culture with 100 ng/mL LPS decreases endothelium-dependent dilation of C...

Organ/body-on-a-chip based on microfluidic technology for drug discovery.

Science.gov (United States)

Kimura, Hiroshi; Sakai, Yasuyuki; Fujii, Teruo

2018-02-01

Although animal experiments are indispensable for preclinical screening in the drug discovery process, various issues such as ethical considerations and species differences remain. To solve these issues, cell-based assays using human-derived cells have been actively pursued. However, it remains difficult to accurately predict drug efficacy, toxicity, and organs interactions, because cultivated cells often do not retain their original organ functions and morphologies in conventional in vitro cell culture systems. In the μTAS research field, which is a part of biochemical engineering, the technologies of organ-on-a-chip, based on microfluidic devices built using microfabrication, have been widely studied recently as a novel in vitro organ model. Since it is possible to physically and chemically mimic the in vitro environment by using microfluidic device technology, maintenance of cellular function and morphology, and replication of organ interactions can be realized using organ-on-a-chip devices. So far, functions of various organs and tissues, such as the lung, liver, kidney, and gut have been reproduced as in vitro models. Furthermore, a body-on-a-chip, integrating multi organ functions on a microfluidic device, has also been proposed for prediction of organ interactions. We herein provide a background of microfluidic systems, organ-on-a-chip, Body-on-a-chip technologies, and their challenges in the future. Copyright © 2017 The Japanese Society for the Study of Xenobiotics. Published by Elsevier Ltd. All rights reserved.

Synthesis, structural analysis, Hirshfeld surface, spectroscopic characterization and, in vitro, antioxidant activity of a novel organic cyclohexaphosphate

Science.gov (United States)

Fezai, Ramzi; Mezni, Ali; Rzaigui, Mohamed

2018-02-01

The new hybrid [4-Cl-2-(CH3)C6H3NH3]6P6O18·2H2O was synthesized under normal conditions of temperature and pressure. Single crystal X-ray diffraction study was used to identify its structure. It revealed that this organic cyclohexaphosphate crystallized in the P 1 bar triclinic space group with a = 10.41 (10) Å b = 10.94 (7) Å, c = 15.45 (10) Å, α = 77.37 (8), β = 89.75 (8)°, γ = 61.69 (7)°, V = 1501 (2) Å3 and Z = 1. In the crystal framework, the assembling of the three dimensional (3D) structure is formed by intermolecular hydrogen bonds and Van Der Waals interactions. A spectroscopic characterization was carried out to elucidate the structure (UV-Vis, FTIR, 31P MAS-NMR and fluorescent properties). The thermal stability was studied by TG-DTA diagrams under argon atmosphere. Furthermore, 3-D Hirshfeld surfaces in combination with 2-D fingerprint plots were carried out. This compound was also evaluated for its antioxidant activity; four tests were done, in vitro, 1,1-diphenyl-2-picrylhydrazyl (DPPH•), hydroxyl scavenging ability (OH•), ferric reducing power (FRP) and ferrous ion chelating (FIC) ability, using ascorbic acid as a control.

Problems in extrapolating genotoxicity data from cellular systems in vitro

Energy Technology Data Exchange (ETDEWEB)

Mohn, G.R.; van Zeeland, A.A.

1986-01-01

The experiments involved quantitative comparative mutagenesis determinations using ethylating agents differing in DNA reactivity, mammalian metabolism, and mutagenic potency. The results can be summarized as follows: (1) Under standardized in vitro treatment conditions and at identical DNA dose levels, the frequencies of induced mutations observed in the present bacterial systems are not representative and usually lower than those induced in cultured mammalian cells. (2) Under the same in vitro treatment conditions, the relative mutagenic potency order is identical in bacteria and the mammalian cells, namely, N-ethyl-N'-nitrosoquinioline(ENNG) > ethylnitrosourea (ENU) > diethylsulfate (DES) > ethylmethanesulfonate (EMS); diethylnitrosamine (DEN) being not mutagenic in the two organisms without addition of mammalian (rodent) liver preparations containing Cyt-P450-dependent mixed function oxidases. (3) The extremely high mutagenic activity of ENNG in bacteria and mammalian cells in vitro is very probably due to its intracellular, glutathione-mediated activation to highly reactive chemical species. (4) Both in bacteria and the mammalian cells the frequencies of induced mutations are directly proportional to the levels of O6-ethylguanine formed in DNA after exposure to ENNG, ENU, DES, and EMS, in contrast to results obtained when other DNA adducts are used as dose parameters. (5) The addition of mouse liver S-9 preparations in the in vitro assays allows to determine which chemicals are likely to be substantially activated (DEN) or deactivated (ENNG) in vivo; however, the mutagenic potency order remained the same under these conditions. (6) When the indicator bacteria are exposed directly to the in vivo metabolism of living animals, the relative order of mutagenic potency on the basis of exposure levels becomes drastically different from that observed in vitro, using either E. coli or V79 mammalian cells.

ENRAIZAMENTO DE CRAVO (Dianthus caryophyllus L. IN VITRO E EX VITRO

Directory of Open Access Journals (Sweden)

G.R.F CUZZUOL

1996-01-01

Full Text Available Plântulas de cravo (Dianthus caryophyllus micropropagadas durante várias gerações pelo período de um ano, foram enraizadas in vitro com AIA, ANA e AIB nas concentrações de 0,0; 0,25; 0,5 e 1,0 mg/l, em fatorial do tipo 3 x 4, com todos os tratamentos promovendo a formação de raízes, mas não diferindo do controle. Foi confrontado em condição autotrófica, o desempenho entre plântulas enraizadas in vitro na presença e ausência do regulador AIA 0,5 mg/l e plântulas enraizadas ex vitro, sem nenhuma diferença quanto ao comprimento da parte aérea. Para a variável produção de massa de matéria seca os melhores resultados foram proporcionados pelas plantas que passaram pela fase de enraizamento in vitro, tendo o sistema radicular efeito sinergístico no crescimento da parte aérea.Plantlets of carnation (Dianthus caryophyllus L. micropropagated through several generations during one year, were observed with respect to rooting in vitro, in the presence of IAA , NAA and IBA, at the following concentrations: 0,0; 0,25; 0,5 and 1,0 mg/l. All treatments promoted root formation, however no differences were detected in comparison to control. As far as the lenght of the aerial part is concerned no difference was observed between in vitro rooting. in the presence or absence of IAA 0,5 mg/l, and ex vitro rooting. Plantlets which were rooted in vitro conditions showed higher production of fresh matter then those rooted ex vitro. The root system had a synergistic effect on the growth of the aerial part.

Characteristic MR spectroscopy in fucosidosis: in vitro investigation

Energy Technology Data Exchange (ETDEWEB)

Mamourian, Alex C.; Chawla, Sanjeev; Poptani, Harish [Hospital of the University of Pennsylvania, Department of Radiology, Philadelphia, PA (United States); Hopkin, Jeremy R. [Barrow Neurological Institute, Department of Neuroradiology, Phoenix, AZ (United States)

2010-08-15

Fucosidosis is a rare lysosomal storage disorder that results in the deposition of the sugar fucose within various organs, including the central nervous system. Neuroimaging abnormalities on MR, specifically T2 shortening in the basal ganglia, have been reported as suggestive of fucosidosis. A more recent report of MR spectroscopy (MRS) of one patient provided evidence that MRS is specific for fucosidosis. We present another confirmed case with nearly identical MR spectroscopic findings along with in vitro data that support the contention that MR spectroscopy, in the setting of typical clinical and imaging features, is characteristic for this rare disorder. (orig.)

Characteristic MR spectroscopy in fucosidosis: in vitro investigation

International Nuclear Information System (INIS)

Mamourian, Alex C.; Chawla, Sanjeev; Poptani, Harish; Hopkin, Jeremy R.

2010-01-01

Fucosidosis is a rare lysosomal storage disorder that results in the deposition of the sugar fucose within various organs, including the central nervous system. Neuroimaging abnormalities on MR, specifically T2 shortening in the basal ganglia, have been reported as suggestive of fucosidosis. A more recent report of MR spectroscopy (MRS) of one patient provided evidence that MRS is specific for fucosidosis. We present another confirmed case with nearly identical MR spectroscopic findings along with in vitro data that support the contention that MR spectroscopy, in the setting of typical clinical and imaging features, is characteristic for this rare disorder. (orig.)

A Simplified Method for Tissue Engineering Skeletal Muscle Organoids in Vitro

Science.gov (United States)

Shansky, Janet; DelTatto, Michael; Chromiak, Joseph; Vandenburgh, Herman

1996-01-01

Tissue-engineered three dimensional skeletal muscle organ-like structures have been formed in vitro from primary myoblasts by several different techniques. This report describes a simplified method for generating large numbers of muscle organoids from either primary embryonic avian or neonatal rodent myoblasts, which avoids the requirements for stretching and other mechanical stimulation.

Synthesis and evaluation of new organic and phosphorous derivatives against ionizing radiation: study of the in vitro mechanism of action

International Nuclear Information System (INIS)

Prouillac, C.

2006-10-01

This work falls under a research program. The aim was to synthesize new organic phosphorylated compounds having an interesting radio pharmacological activity without toxicity. That is why, we carried out the synthesis of new benzothiazole and thiadiazole N-substituted derivatives as thiols, amino thiols, acids thio-sulfonic and phosphoro thioates. All these compounds were characterized by NMR (proton, carbon, phosphorus, 2D), by mass spectrometry, elementary analyzes and for some of them by diffraction of x-rays. The activity of the majority of them was evaluated by in vitro tests. The experimental results were confirmed by theoretical study: the aim of D.F.T. calculation was the study of the mechanism of capture of the free radicals by our compounds. In addition, a study of relation structure activity (Q.S.A.R.) was carried out. Our results allow us to create a model making it possible to establish structure-activity relationship. (author)

CHLAMYFAST-OIA TEST IN THE GENITAL CHLAMYDIA MALE INFECTION DIAGNOSIS

Directory of Open Access Journals (Sweden)

Anka Vasic

2004-07-01

Full Text Available The genital infections caused by Chlamydia trachomatis (Ch. trachomatis, Mycoplasma hominis (M. hominis and Ureaplasma urealyticum (U. urealyticum represent, in the countries with developed industry, those diseases which are most often sexually transmissible. Chronic infections provoked by the mentioned causes are considered to be the risk factors for sterility.The aim of this paper is to examine the importance and specific characteristics of the CHLAMYFAST-OIA test in the Chlamydia genital infection diagnosis. This study includes 400 male patients with urethritis symptoms. The CHLAMYFAST-optical immunologic test has been used to determine the presence of the Ch. trachomatis in the genital tract of 360 males (Mycoplasma, International, France. The genital microplasmas, that is M. hominis and U. urealyticum, have been detected with the use of MYCOFAST-test (Mycroplasm International, France. The presence of the genital microplasmas has been studied in 129 patients.Chlamydia genital infection has been determined in 128 males (35,55%. The genital infection caused by M. hominis has been determined in a largely lower number of patients (3; 2,32%, as well as the infection caused by U. urealyticum (in 8 patients; 6,20%. Mixed infections have been detected in 8 patients. In 6 men (4,64% there has been detected a mixed infection caused by genital microplasmas. The mixed infection provoked by Ch. Trachomatis and M. hominis, and the one caused by Ch. trachomatis and U. urealyticum, has been proven only in one patient respectively.

In vitro culture of higher plants as a tool in the propagation of horticultural crops.

NARCIS (Netherlands)

Pierik, R.L.M.

1988-01-01

In vitro culture of higher plants is the culture, under sterile conditions, of plants, seeds, embryos, organs, explants, tissues, cells and protoplasts on nutrient media. This type of culture has shown spectacular development since 1975, resulting in the production and regeneration of viable

In vitro remineralization of in vivo and in vitro formed enamel lesions

NARCIS (Netherlands)

Iijima, Y; Takagi, O; Ruben, J; Arends, J

1999-01-01

Thin sections of natural white spot enamel lesions (WS) and of artificial in vitro lesions (VL) were remineralized simultaneously in vitro. The sections, clamped in a PMMA holder, were microradiographed at baseline and after remineralization in a calcium- and phosphate-containing solution (pH = 7.0;

Quantitative Analysis of Nanoparticle Transport through in Vitro Blood-Brain Barrier Models

NARCIS (Netherlands)

Åberg, Christoffer

2016-01-01

Nanoparticle transport through the blood-brain barrier has received much attention of late, both from the point of view of nano-enabled drug delivery, as well as due to concerns about unintended exposure of nanomaterials to humans and other organisms. In vitro models play a lead role in efforts to

Analysis of Chemical Bioactivity through In Vitro Profiling ...

Science.gov (United States)

Safety assessment of drugs and environmental chemicals relies extensively on animal testing. However, the quantity of chemicals needing assessment and challenges of species extrapolation drive the development of alternative approaches. The EPA’s ToxCast and the multiagency Tox21 programs address this through use of an extensive in vitro screening program to generate data on a large library of important environmental chemicals. These in vitro assays encompass both cell-free, biochemical assays targeting proteins that may be potential molecular initiating events and cellular assays that provide coverage of critical signaling pathways and toxicity phenotypes. Effects on model organisms such as the developing zebrafish, are also part of the testing strategy. A variety of computational approaches are used to analyze the resulting complex data sets to gain insight in to inherent biological activity of chemicals and possible mechanisms of toxicity. Several case studies including identification of modulators of estrogen receptor and aromatic hydrocarbon receptor pathways with effects in primary human cell systems will be described. In addition, existing in vivo data from a subset of the chemicals was used to anchor predictive models using in vitro data for a number of adverse endpoints including reproductive and developmental toxicities. The strengths and weaknesses of this approach will be described. This work does not necessarily reflect official Agency policy. Pres

Study in vitro of origin radioprotective food the radioprotective effect in vitro of food borne; Estudio in vitro de radioprotectores de origen alimentario

Energy Technology Data Exchange (ETDEWEB)

Soraino, J. M.; Sebastia, N.; Almonacid, M.; Alonso, O.; Cervera, J.; Such, E.; Silla, M. A.; Villaescusa, J. I.; Montoro, A.

2012-07-01

Study in vitro of origin radioprotective food the radioprotective effect in vitro of food borne substances studied is a first step in developing effective radioprotectors that can prevent radiation damage to healthy tissue., cannot forget that these studies must be accompanied by in vitro studies of toxicity and bioavailability to profile designing radioprotective substance.

Anatomia comparada das folhas e raízes de Cymbidium Hort. (Orchidaceae cultivadas ex vitro e in vitro Comparative leaf and root anatomy of ex vitro and in vitro cultured Cymbidium Hort. plants

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Juliana Lischka Sampaio Mayer

2008-06-01

Full Text Available Na fase de cultivo in vitro, as plantas são mantidas em ambiente com alta umidade relativa do ar, baixa luminosidade e trocas gasosas restritas, o que resulta em taxa de transpiração reduzida. Portanto, quando essas mudas são expostas ao meio ex vitro, sofrem estresse que pode causar a morte. O objetivo desse trabalho foi comparar a estrutura anatômica das mudas de Cymbidium 'Joy Polis' cultivadas ex vitro (planta matriz e aclimatizada e in vitro e verificar se a estrutura anatômica das plantas in vitro influencia no processo de aclimatização. As plantas ex vitro foram mantidas em casa-de-vegetação, em vasos individuais com o substrato fibra de coco em pó combinada com fibra de coco, e as plantas in vitro foram mantidas em meio de cultura MS. Para a análise anatômica qualitativa foram coletadas amostras de folhas e raízes de plantas ex vitro e in vitro. As plantas aclimatizadas apresentaram estrutura morfoanatômica semelhante à da planta matriz. A estrutura anatômica das plantas in vitro não influenciou a sobrevivência das mudas durante a aclimatização devido à plasticidade fenotipica desse cultivar. As plantas de Cymbidium 'Joy Polis' possuem grande capacidade de aclimatização ao ambiente, sendo provavelmente este um dos fatores responsáveis pela sobrevivência de 100% das mudas.During in vitro culture plants are kept in an atmosphere with high relative humidity, low light intensity and reduced gas exchange, resulting in low transpiration rates. Therefore, when these plants are exposed to ex vitro conditions, they suffer stress, which can induce mortality. The purpose of this study was to compare the anatomical structure of Cymbidium 'Joy Polis' plants from ex vitro (mother plant and acclimatized plants and in vitro cultures and to verify if the anatomical structure of in vitro cultured plants affects acclimatization. The ex vitro plants were kept in a greenhouse in pots containing a mixture of coconut-fiber powder and

In Vitro Mimetic Models for the Bone-Cartilage Interface Regeneration.

Science.gov (United States)

Bicho, Diana; Pina, Sandra; Oliveira, J Miguel; Reis, Rui L

2018-01-01

In embryonic development, pure cartilage structures are in the basis of bone-cartilage interfaces. Despite this fact, the mature bone and cartilage structures can vary greatly in composition and function. Nevertheless, they collaborate in the osteochondral region to create a smooth transition zone that supports the movements and forces resulting from the daily activities. In this sense, all the hierarchical organization is involved in the maintenance and reestablishment of the equilibrium in case of damage. Therefore, this interface has attracted a great deal of interest in order to understand the mechanisms of regeneration or disease progression in osteoarthritis. With that purpose, in vitro tissue models (either static or dynamic) have been studied. Static in vitro tissue models include monocultures, co-cultures, 3D cultures, and ex vivo cultures, mostly cultivated in flat surfaces, while dynamic models involve the use of bioreactors and microfluidic systems. The latter have emerged as alternatives to study the cellular interactions in a more authentic manner over some disadvantages of the static models. The current alternatives of in vitro mimetic models for bone-cartilage interface regeneration are overviewed and discussed herein.

Reporting detection of Chlamydia trachomatis DNA in tissues of neonatal death cases

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Maria Hernandez Trejo

2014-04-01

Full Text Available OBJECTIVE: to determine whether C. trachomatis was present in neonates with infection, but without an isolated pathogen, who died during the first week of life. METHODS: early neonatal death cases whose causes of death had been previously adjudicated by the institutional mortality committee were randomly selected. End-point and real-time polymerase chain reaction of the C. trachomatis omp1 gene was used to blindly identify the presence of chlamydial DNA in the paraffinized samples of five organs (from authorized autopsies of each of the dead neonates. Additionally, differential diagnoses were conducted by amplifying a fragment of the 16S rRNA of Mycoplasma spp. RESULTS: in five cases (35.7%, C. trachomatis DNA was found in one or more organs. Severe neonatal infection was present in three cases; one of them corresponded to genotype D of C. trachomatis. Interestingly, another case fulfilled the same criteria but had a positive polymerase chain reaction for Mycoplasma hominis, a pathogen known to produce sepsis in newborns. CONCLUSION: the use of molecular biology techniques in these cases of early infant mortality demonstrated that C. trachomatis could play a role in the development of severe infection and in early neonatal death, similarly to that observed with Mycoplasma hominis. Further study is required to determine the pathogenesis of this perinatal infection.

Pengembangan pemakaian hasil samping agroindustri berbahan dasar jagung sebagai alternatif bahan baku pakan ikan kerapu tikus Cromileptes altivelis

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Muhammad Agus Suprayudi

2015-03-01

Full Text Available ABSTRACTThis experiment was conducted to evaluate the use of by product of corn ethanol industry, DDGS (distiller dried grain with soluble and hominy as feed raw material on digestibility and the growth performance of humpback grouper Cromileptes altivelis juvenile. Three experimental diets containing isoprotein (48% but different in DDGS contain were used. Diet A containing 0% of DDGS and hominy while diet B and C containing 10.13% (6.13% of DDGS and 4% of hominy and 19.05% (15.05% of DDGS and 4% of hominy. Reduce of fish meal level in the diet was happen as a consequence of balancing amino acid. For digestibility experiment and Cr2O3 at level of 0.05% was used as a tracer. The feces were daily collected 30‒60 minutes after feeding for 14 days. Dry matter and protein were used as digestibility parameters. Completely randomized design with three treatments and two replicates was used in this experiment. Humpback grouper with an average weight of 11‒14 g were reared at floating net cages (FNG of 3×2×1.5 m3. The stocking density for each cage was 167 fish/FNG and reared for six months. For digestibility measurement 30 fish were held in cylindrical fiber tank filled with 800 L of sea water. Fish were fed three times daily at satiation level. For maintaining net in a clean condition net was changed in every ten days. Feed consumption, digestibility of protein and total, growth performance indicator i,e specific growth rate, feed efficiency protein and lipid retention were used as evaluating parameters. The results showed that the fish fed with diet containing DDGS (6.13% dan hominy (4%, at the level of 10.13% (diet B, shows similar growth performance indicators with the fish fed diet A (0% of DDGS and hominy and the growth performance indicator was higher when being compared to those fish fed diet C (containing 19.05% of DDGS (15.05% and hominy (4% (p<0.05. More over the inclusion of DDGS and hominy at the level of 10,13% give no negatif

In vivo and in vitro study of /sub 90/Sr in developing rat molar enamel

International Nuclear Information System (INIS)

White, B.A.; Deaton, T.G.; Bawden, J.W.

1980-01-01

The uptake patterns of /sub 90/Sr in developing rat molar enamel were studied in vivo and in vitro. Autoradiographic methods were used that preclude loss or translocation of tracers associated with water-soluble compounds in the sections. In eight-day-old rats injected with the tracer, /sub 90/Sr uptake in the enamel was significantly less than for dentin and bone, particularly at early sacrifice times. The uptake pattern of 90Sr was somewhat different from that previously observed for /sub 45/Ca. The in vitro experiments indicated that the viable intact enamel organ limits uptake of /sub 90/Sr by enamel in both the secretory and maturation phases of enamel formation

In Vitro Digestibilities of Six Rumen Protected Fat-Protein Supplement Formulas

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Lilis Hartati

2012-01-01

Full Text Available Abstract. The aim of the research was to evaluate the efficacy of protection method of rumen protected fat-protein supplements. In vitro digestibility test was carried out to examine nutrients digestibility of different supplement formula based on the sources of protein and oil. The research used two sources of fat namely crude palm oil (CPO and fish oil (FO and three sources of protein namely milk skim, soy flour, and soybean meal. Thus there were 6 combinations that subjected in the in vitro digestibility test. The observed variables were the digestibility of dry matter (DM, organic matter (OM, crude fat (CF, and crude protein (CP. Results indicated that the method for protecting protein and fat was effective. This was showed by low nutrients digestibility in the rumen and high nutrients digestibility in the post rumen. In conclusion the combination between skim milk and CPO gave the best results among the other supplement formula. Keywords: rumen protected nutrient, fat-protein supplement, rumen digestibility, in vitro Animal Production 14(1:1-5, January 2012

NUTRITIONAL EVALUATION OF VARIOUS FEEDSTUFFS FOR LIVESTOCK PRODUCTION USING IN VITRO GAS METHOD

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S. A. KHANUM, T. YAQOOB1, S. SADAF1, M. HUSSAIN, M. A. JABBAR1, H. N. HUSSAIN, R. KAUSAR AND S. REHMAN1

2007-07-01

Full Text Available A study was undertaken to evaluate the nutritional quality of some conventional and non-conventional feed resources by using in vitro gas method. Samples of various feedstuffs were analyzed chemically, as well as by in vitro gas method. The feedstuffs having different digestibilities showed significant (P<0.05 differences in the rate and amount of gas production, metabolizable energy (ME and digestibility of organic matter. Predicted metabolizable energy values were very low in feedstuffs having high fiber and low protein contents. These feedstuffs included various grasses, crop residues and wheal straw. Lowest ME value of 4.7 MJ/kg of dry matter (DM was found in wheat straw. Many of the roughages (Sorghum vulgare, Kochia indica, Leptochloa fusca studied were found to be deficient in fermentable carbohydrates, resulting in low organic matter digestibility. Concentrate feed stuffs like cotton seed meal, sunflower meal, cotton seed cakes, rice polish, rapeseed meal and Zea mays (maize grains had higher ME values (9.27 – 12.44 MJ/kg DM. The difference of ME of various feedstuffs reflects different contents of fermentable carbohydrates and available nitrogen in cereals and protein supplements. Among the non-conventional feedstuffs, Acacia ampliceps, Acacia nilotica, Sesbania aculeata, Leptochloa fusca and Prosopis juliflora were found potential fodders. Extensive use of in vitro gas method proved its potential as a tool to evaluate various ruminant feeds for energy component.

Bioaccessibility of polychlorinated biphenyls in different foods using an in vitro digestion method

International Nuclear Information System (INIS)

Xing Guanhua; Yang Yu; Chan, Janet Kie Yan Chan; Tao Shu; Wong Minghung

2008-01-01

Bioaccessibility of organic pollutants in food is important for exposure estimation and risk assessment. An in vitro enzyme digestion experiment was carried out to analyze the bioaccessibility of PCBs in different foods including freshwater fish (bighead carp and oriental weather fish) and vegetables (spinach and cabbage). The results indicated that the bioaccessibility of PCBs in freshwater fish (3%) was much lower than that in leafy vegetables (25%). Based on field survey, the daily exposure TEQ value from these two types of food was 17.9 pg/kg bw/day. However, when bioaccessibility was taken into consideration, the value decreased to 0.61 pg/kg bw/day. Therefore, the forms of PCBs existing in food strongly influenced the bioavailability for humans, which may have important implications on dietary exposure. To our knowledge, this is the first paper to illustrate the bioaccessibility of PCBs in different foods using an in vitro digestion method. - This paper describes the applicability of an in vitro digestion method for measuring the bioaccessibility of PCBs from freshwater fish and leafy vegetables

Low-temperature bonded glass-membrane microfluidic device for in vitro organ-on-a-chip cell culture models

Science.gov (United States)

Pocock, Kyall J.; Gao, Xiaofang; Wang, Chenxi; Priest, Craig; Prestidge, Clive A.; Mawatari, Kazuma; Kitamori, Takehiko; Thierry, Benjamin

2015-12-01

The integration of microfluidics with living biological systems has paved the way to the exciting concept of "organson- a-chip", which aims at the development of advanced in vitro models that replicate the key features of human organs. Glass based devices have long been utilised in the field of microfluidics but the integration of alternative functional elements within multi-layered glass microdevices, such as polymeric membranes, remains a challenge. To this end, we have extended a previously reported approach for the low-temperature bonding of glass devices that enables the integration of a functional polycarbonate porous membrane. The process was initially developed and optimised on specialty low-temperature bonding equipment (μTAS2001, Bondtech, Japan) and subsequently adapted to more widely accessible hot embosser units (EVG520HE Hot Embosser, EVG, Austria). The key aspect of this method is the use of low temperatures compatible with polymeric membranes. Compared to borosilicate glass bonding (650 °C) and quartz/fused silica bonding (1050 °C) processes, this method maintains the integrity and functionality of the membrane (Tg 150 °C for polycarbonate). Leak tests performed showed no damage or loss of integrity of the membrane for up to 150 hours, indicating sufficient bond strength for long term cell culture. A feasibility study confirmed the growth of dense and functional monolayers of Caco-2 cells within 5 days.

Evaluation of the Roche LightMix Gastro parasites multiplex PCR assay detecting Giardia duodenalis, Entamoeba histolytica, cryptosporidia, Dientamoeba fragilis, and Blastocystis hominis.

Science.gov (United States)

Friesen, J; Fuhrmann, J; Kietzmann, H; Tannich, E; Müller, M; Ignatius, R

2018-03-23

Multiplex PCR assays offer highly sensitive and specific tools for the detection of enteric pathogens. This prospective study aimed at comparing the novel Roche LightMix Modular Assay Gastro Parasites (LMAGP) detecting Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Blastocystishominis, and Dientamoebafragilis with routine laboratory procedures. Stool specimens (n = 1062 from 1009 patients) were consecutively examined by LMAGP, R-Biopharm Ridascreen enzyme immunoassays (EIAs) detecting G. duodenalis or E. histolytica/dispar, and microscopy of wet mounts. Discrepant results were analysed by in-house PCR. D. fragilis or B. hominis were detected by LMAGP in 131 (14.4%) and 179 (19.9%; 16 samples positive by microscopy; p PCR). G. duodenalis was detected by LMAGP, EIA, or microscopy in 20, 16, or 9 of 1039 stool samples, respectively; all four samples missed by EIA were confirmed by in-house PCR. In total, 938 stool samples were analysed for E. histolytica/dispar. Nine of ten EIA-positive samples were negative by LMAGP but positive by in-house PCR for E. dispar. One E. histolytica infection (positive by both LMAGP and in-house PCR) was missed by EIA and microscopy. Parasites only detected by microscopy included Enterobius vermicularis eggs (n = 3) and apathogenic amoebae (n = 27). The data call for routine use of multiplex PCR assays for the detection of enteric protozoan parasites in laboratory diagnostics. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Prospective Clinical Testing of Regulatory Dendritic Cells in Organ Transplantation

OpenAIRE

Thomson, Angus W.; Zahorchak, Alan F.; Ezzelarab, Mohamed B.; Butterfield, Lisa H.; Lakkis, Fadi G.; Metes, Diana M.

2016-01-01

Dendritic cells (DC) are rare, professional antigen-presenting cells with ability to induce or regulate alloimmune responses. Regulatory DC (DCreg) with potential to down-modulate acute and chronic inflammatory conditions that occur in organ transplantation can be generated in vitro under a variety of conditions. Here, we provide a rationale for evaluation of DCreg therapy in clinical organ transplantation with the goal of promoting sustained, donor-specific hyporesponsiveness, while lowering...

Genital mycoplasmas in semen samples of males attending a tertiary care hospital in Nigeria: any role in sperm count reduction?

Science.gov (United States)

Agbakoba, N R; Adetosoye, A I; Ikechebelu, J I

2007-06-01

Semen samples from 54 married men attending the outpatient clinics for problems of infertility and routine semen analysis were examined for the presence of genital mycoplasmas. The mean age of the men was 36.1 years with a range of 25 55 years. Majority of the men 57.4% (31 of 54) were in their fourth decade of life (30 39 years). This age group also had the highest percentage 57.2% (8 of 14) of positive isolates of genital mycoplasmas on semen culture. A total of 21 organisms obtained from 14 (26.0%) positive samples were isolated. Mycoplasma and Ureaplasma spp. separately isolated from the samples yielded frequencies of 1 (1.9%) and 6 (11.1%) respectively and the remaining 7 (13.0%) samples were infected with both organisms. A breakdown of the mycoplasma species include 5 (23.8%) M. hominis, 2 (9.5%) M. fermentans and 1 (4.8%) M. penetrans. Apart from one isolate of M. hominis other Mycoplasma species were found in association with Ureaplasma species. Fifteen (71.4%) of the 21 isolates [8 (53.3%) ureaplasmas and 7 (46.7%) mycoplasmas] were isolated from samples with sperm counts less than 20 million/ml while the remaining 6 (21.6%) isolates [5 (83.3%) ureaplasmas and 1 (16.7) mycoplasma] were from samples with counts greater than 20 million/ml. This finding could indicate a possible influence of genital mycoplasmas especially mycoplasmas species on sperm count.

Effect of Formic Acid on In Vitro Ruminal Fermentation and Methane Emission

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Kanber Kara

2015-10-01

Full Text Available In this study, it was aimed to investigate the effects of formic acid on the in vitro methane production and in vitro ruminal fermentation of alfalfa hay. Effect of 0.0 (control group: YF0, 0.1, 0.2, 0.3, 0.4 and 0.5 ml/L (experimental groups: YF1, YF2, YF3, YF4, and YF5 respectively formic acid (Amasil85-liquid addition to rumen fluid on ruminal fermentation parameters of alfalfa hay were determined by using in vitro gas production techniques. Methane production of in vitro incubation increased (to about 20% with addition of linearly increased formic acid. Linearly increased levels of formic acid addition to rumen fluid has significantly changed the production of in vitro total gas production, metabolic energy (ME and organic matter digestibility (OMD at linear, quadratic and cubic. The addition of 0.1 ml/L and 0.2 ml/L formic acid to rumen fluid significantly decreased in vitro total gas production, ME and OMD however addition of 0.3 ml/L and 0.4 ml/L formic acid was not changed in vitro gas production, ME and OMD levels and 0.5 ml/L formic acid was significantly increased all these parameters. Ruminal pH was not changed by addition of formic acid. Formic acid is a safe feed additive because of its properties antibacterial and flavorings and also is used as a fermentation promoter in silage. In this study it has been observed that all doses of formic acid increased in vitro enteric methane production and low doses decreased in vitro total gas production, ME and OMD and high doses have increased all these parameters. High doses have a positive effect on ME and OMD; however formic acid should be used at limited levels in diets due to the negative effect of increasing greenhouse gases. The effect of formic acid addition to the feed raw matter and rations of all livestock would be beneficial to investigate in terms of digestive system parameters and global warming, further in vitro and in vivo studies.

Sildenafil vaginal suppositories: preparation, characterization, in vitro and in vivo evaluation.

Science.gov (United States)

Shanmugam, Srinivasan; Kim, Young-Hun; Park, Jeong-Hee; Im, Ho Taek; Sohn, Young Taek; Kim, Kyeong Soo; Kim, Yong-Il; Yong, Chul Soon; Kim, Jong Oh; Choi, Han-Gon; Woo, Jong Soo

2014-06-01

The main objective was to investigate the in vitro release profile/kinetics, and in vivo plasma pharmacokinetics (PK) and organ biodistribution (BD) of the prepared sildenafil vaginal suppositories (SVS). Suppositories containing 25 mg of sildenafil were prepared by the cream melting technique using Witepsol H-15 as a suppository base. The suppositories were characterized for weight variation, content uniformity, hardness, disintegration time and crystallinity change. The in vitro dissolution in pH 4.5, and in vivo plasma PK and organ BD of sildenafil from SVS in female Sprague Dawley rats, were also investigated. The mean weight variation, content uniformity, hardness and disintegration time of the prepared SVS were 1.127 ± 0.020 g, 98.25 ± 2.50%, 2.5 ± 0.08 kg and 9 ± 1.0 min, respectively. The release of sildenafil from the SVS was more than 90% at 30 min, with a release kinetic of Hixson--Crowell model and non-Fickian diffusion (n = 0.464). The plasma PK study demonstrated a significantly lower Cmax (∼10 times) and AUC0-24 h (∼13 times) of sildenafil in plasma following intravaginal (IVG) administration of suppositories compared to oral (PO) administration of sildenafil solution. Nevertheless, the organ BD study showed a phenomenally higher Cmax (∼40 times) and AUC0-24 h (∼20 times) of sildenafil in uterus following IVG administration of suppositories than PO administration of sildenafil solution. This study demonstrated enhanced sildenafil exposure in the uterus following IVG administration of SVS, which could be used to target the uterus for therapeutic benefits.

In vitro action of 1,25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol on matrix organization and mineral distribution in rabbit growth plate.

Science.gov (United States)

Plachot, J J; Du Bois, M B; Halpern, S; Cournot-Witmer, G; Garabedian, M; Balsan, S

1982-01-01

Growth plates of 18-day-old rabbits were incubated in a protein-free synthetic medium, either without any additive, with 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] (10(-10) M), with 24,25-dihydroxycholecalciferol [24,25-(OH)2D3] (10(-10) M and 10(-9) M), with both metabolites, or with the ethanol solvent alone. Cartilages, before and after 5 days of incubation, were studied by light and electron microscopy. The intracellular calcium distribution was analyzed by the potassium pyroantimonate method, and the calcium content was verified by x-ray microprobe analysis. When compared to nonincubated samples the cartilages incubated for 5 days without any additive as well as the cartilages incubated with the solvent alone showed excessive hydratation and hypertrophy of the chondrocytes, which had lost their columnar arrangement. The matrix and the cells were devoid of mineral. The ultrastructure of the cells was well preserved. These changes were largely prevented by the presence of both vitamin D3 metabolites. With regard to calcium distribution, 1,25-(OH)2D3 maintained calcium in mitochondria and crystals in matrix vesicles, whereas 24,25-(OH)2D3 only partly maintained mitochondrial mineral. In the chondrocytes incubated with this latter metabolite, small calcium granules were seen in the cytoplasm; most vesicles were devoid of crystals, and amorphous precipitates were seen in the matrix. These data demonstrate the in vitro influence of vitamin D3 metabolites on the organization and mineralization of the cartilage matrix and on the distribution of intracellular calcium in chondrocytes. Furthermore, they support the hypothesis that the in vitro action of 1,25-(OH)2D3 is different from that of 24,25-(OH)2D3 in that 1,25-(OH)2D3 may influence calcium storage in mitochondria and matrix vesicles, whereas 24,25-(OH)2D3 is likely to be involved in calcium transport and release.

Relationship between in situ degradation kinetics and in vitro gas production fermentation using different mathematical models

NARCIS (Netherlands)

Rodrigues, M.A.M.; Cone, J.W.; Ferreira, L.M.M.; Blok, M.C.; Guedes, C.

2009-01-01

In vitro and in situ studies were conducted to evaluate the influence of different mathematical models, used to fit gas production profiles of 15 feedstuffs, on estimates of nylon bag organic matter (OM) degradation kinetics. The gas production data were fitted to Exponential, Logistic, Gompertz and

A study on volatile organic compounds emitted by in-vitro lung cancer cultured cells using gas sensor array and SPME-GCMS.

Science.gov (United States)

Thriumani, Reena; Zakaria, Ammar; Hashim, Yumi Zuhanis Has-Yun; Jeffree, Amanina Iymia; Helmy, Khaled Mohamed; Kamarudin, Latifah Munirah; Omar, Mohammad Iqbal; Shakaff, Ali Yeon Md; Adom, Abdul Hamid; Persaud, Krishna C

2018-04-02

Volatile organic compounds (VOCs) emitted from exhaled breath from human bodies have been proven to be a useful source of information for early lung cancer diagnosis. To date, there are still arguable information on the production and origin of significant VOCs of cancer cells. Thus, this study aims to conduct in-vitro experiments involving related cell lines to verify the capability of VOCs in providing information of the cells. The performances of e-nose technology with different statistical methods to determine the best classifier were conducted and discussed. The gas sensor study has been complemented using solid phase micro-extraction-gas chromatography mass spectrometry. For this purpose, the lung cancer cells (A549 and Calu-3) and control cell lines, breast cancer cell (MCF7) and non-cancerous lung cell (WI38VA13) were cultured in growth medium. This study successfully provided a list of possible volatile organic compounds that can be specific biomarkers for lung cancer, even at the 24th hour of cell growth. Also, the Linear Discriminant Analysis-based One versus All-Support Vector Machine classifier, is able to produce high performance in distinguishing lung cancer from breast cancer cells and normal lung cells. The findings in this work conclude that the specific VOC released from the cancer cells can act as the odour signature and potentially to be used as non-invasive screening of lung cancer using gas array sensor devices.

Evaluation of the toxicity of organic matter in marine sediments

Digital Repository Service at National Institute of Oceanography (India)

Sarkar, A.

The inhibitory effects of the organics from the sediment along the east coast of India on acetylcholinesterase (AChE) in-vitro were evaluated. Sediment samples collected from the surface layers at various depths were extracted. Each of the extracts...

Regeneration of some monocotyledonous plants from subterranean organs in vitro

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Krystyna D. Kromer

2013-12-01

Full Text Available The aim in view was investigation ofthe regenerative potential of rhizomes, bulbs and corms of ten monocotyledonous plant species from four families: Amaryllidaceae (Haemanthus katharinae, Crinum abyssinicum, Leucojum vernum, Araceae (Spathiphyllum wallisii, Iridaceae (Crocus vernus, Iris germanica, Liliaceae (Hosta lancifolia, Muscari racemosum, Scilla laxiflora, Veltheimia viridifolia under conditions of in vitro culture. All the investigated species were capable of buld or, alternatively, bud and root regeneration. Different morphogenetic potential was noted between the particular families. A high regenerative potential under the conditions of culture applied was characteristic for plants of the Liliaceae and Araceae families, it was lower in plants belonging to Amaryllidaceae and lowest in those of the Iridaceae family. Plants from the Liliaceae family exhibited also the highest ability of callus formation, whereas Amaryllidaceae and Iridaceae plants possessed this ability in only a low degree. The influence of growth regulators of the auxin group (NAA, IAA, 2,4-D and of cytokinins (K on the initiation and course of organogenesis was tested. The results of the experiments indicate that auxins in interaction with kinetin gave the highest percentage of regenerating explants and also a large number of buds on the latter. Stimulation of callus tissue was highest under the influence of 2,4-D, and weaker when NAA IAA were used.

Acclimatization of in Vitro-derived Dendrobium

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Jaime A. Teixeira da Silva

2017-05-01

Full Text Available The successful ex vitro establishment of Dendrobium plantlets raised in vitro determines the quality of the end product (cut flowers or potted plants in commercial production for economic gain. When in vitro Dendrobium plantlets are transplanted from the culture room to greenhouse conditions, they may desiccate or wilt rapidly and can die as a result of changes in the environment, unless substantial precautions are taken to adapt plantlets to a new environment. The acclimatization of in vitro-grown Dendrobium plantlets to an ex vitro environment by gradually weaning them towards ambient relative humidity and light levels facilitates better survival of young and physiologically sensitive plantlets. Dendrobium plantlets raised in vitro must thus undergo a period of acclimatization or transitional development to correct anatomical abnormalities and to enhance their physiological performance to ensure survival under ex vitro conditions. The most common approach to improve the survival of Dendrobium plantlets upon transfer to an ex vitro environment is their gradual adaptation to that environment. Under such conditions, plants convert rapidly from a heterotrophic or photomixotrophic state to an autotrophic growth, develop a fully functional root system, and better control their stomatal and cuticular transpiration. Gradual adaptation is carried out in a greenhouse by decreasing relative humidity using fog or mist chambers and by increasing light intensity using shading techniques. This review details the acclimatization and ex vitro survival of Dendrobium plants produced in vitro. This advice is also useful for other orchids.

Molecular epidemiology of Staphylococcus spp. contamination in the ward environment: study on mecA and femA genes in methicillin-resistant strains.

Science.gov (United States)

Ashimoto, A; Hamada, T; Adachi, A; Tanigawa, T; Tanaka, Y

1995-01-01

Staphylococcus spp. were isolated from the ward environment and antibiotic susceptibility tests were performed. Twenty-nine strains out of 274 isolates were S. aureus, and 41.4% of the S. aureus strains were methicillin resistant (MRSA). All 12 strains of MRSA were also resistant to oxacillin, ceftizoxime, ampicillin and clindamycin. Among the coagulase-negative staphylococci (CNS), methicillin-resistant (MR) strains of S. epidermidis, S. capitis, S. warneri, S. haemolyticus, S. hominis, S. auricularis, S. saprophyticus and S. cohnii were isolated. Eight of the 10 S. Haemolyticus strains were methicillin resistant. The femA gene was detected in S. aureus (MSSA and MRSA), but not in CNS by polymerase chain reaction (PCR) analysis and Southern blot analysis. The mecA gene was found in all the MRSA and MR-S. epidermidis strains tested, and one of the two MR-S. hominis strains, but not in MSSA, MS-S. epidermidis, MS-S. hominis, or MS-S. haemolyticus. DNA from one strain of MR-S. hominis and 2 strains of MR-S. haemolyticus was not amplified by PCR using the mecA gene primer, or hybridized by Southern blotting. The ambiguity that mecA was detected in some MR-CNS strains, but not in others is discussed.

Development in Assay Methods for in Vitro Antimalarial Drug Efficacy Testing: A Systematic Review

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Shweta Sinha

2017-10-01

Full Text Available The emergence and spread of drug resistance are the major challenges in malaria eradication mission. Besides various strategies laid down by World Health Organization, such as vector management, source reduction, early case detection, prompt treatment, and development of new diagnostics and vaccines, nevertheless the need for new and efficacious drugs against malaria has become a critical priority on the global malaria research agenda. At several screening stages, millions of compounds are screened (1,000–2,000,000 compounds per screening campaign, before pre-clinical trials to select optimum lead. Carrying out in vitro screening of antimalarials is very difficult as different assay methods are subject to numerous sources of variability across different laboratories around the globe. Despite this, in vitro screening is an essential part of antimalarial drug development as it enables to resource various confounding factors such as host immune response and drug–drug interaction. Therefore, in this article, we try to illustrate the basic necessity behind in vitro study and how new methods are developed and subsequently adopted for high-throughput antimalarial drug screening and its application in achieving the next level of in vitro screening based on the current approaches (such as stem cells.

Effect of gallium-arsenic laser on photosensitized periodontopathic anaerobic organisms: An in vitro study

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Mathur, Setu; Kothiwale, Shaila Veerappa; Nag, Buddhi Prakash; Mathur, Tanu; Bhansali, Ashoka; Khatri, Rohit Kumar

2016-01-01

Background: The mainstay of periodontal therapy is mechanical removal of subgingival plaque. There is considerable interest in supplementing it with the use of antibiotics and antiseptics. Many drawbacks are associated with these adjunctive pharmacological regimens such as development of resistance to antibiotics and disruption of microflora of the gastrointestinal tract. Hence, alternate means of killing subgingival bacteria are clearly desirable. One such method is the use of laser. Aim: This study aimed to investigate antibacterial capabilities of gallium-arsenic (Ga-As) laser on photosensitized periodontopathic organisms. The three bacteria selected for the study were Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. Settings: The subjects for the study were selected from the patients visiting the Department of Periodontics, Karnataka Lingayat Education Society's Institute of Dental Sciences, Belgaum. Design: In vitro study design. Materials and Methods: Subgingival plaque samples collected from chronic periodontitis patients were cultured anaerobically for 72 h. Predetermined number of colonies of each bacterium was taken and was then divided into cases and control groups. Both groups were photosensitized using toluidine blue O (TBO) dye and the case groups were irradiated with Ga-As laser. Bacterial colonies were then serially diluted and were incubated for subculture. After incubation period, the number of viable bacterial count was performed. Statistical Analysis: Wilcoxon-signed rank test was carried out to determine significance of reduction on subsequent dilution within the bacterial group. Mann–Whitney U-test was performed to determine the significance of reduction between cases and control of particular bacterial group. Results: The results revealed substantial reduction in the viable bacterial count. F. nucleatum was found to be most sensitive to killing by laser irradiation followed by P. intermedia and then P

Effect of gallium-arsenic laser on photosensitized periodontopathic anaerobic organisms: An in vitro study

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Setu Mathur

2016-01-01

Full Text Available Background: The mainstay of periodontal therapy is mechanical removal of subgingival plaque. There is considerable interest in supplementing it with the use of antibiotics and antiseptics. Many drawbacks are associated with these adjunctive pharmacological regimens such as development of resistance to antibiotics and disruption of microflora of the gastrointestinal tract. Hence, alternate means of killing subgingival bacteria are clearly desirable. One such method is the use of laser. Aim: This study aimed to investigate antibacterial capabilities of gallium-arsenic (Ga-As laser on photosensitized periodontopathic organisms. The three bacteria selected for the study were Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. Settings: The subjects for the study were selected from the patients visiting the Department of Periodontics, Karnataka Lingayat Education Society's Institute of Dental Sciences, Belgaum. Design: In vitro study design. Materials and Methods: Subgingival plaque samples collected from chronic periodontitis patients were cultured anaerobically for 72 h. Predetermined number of colonies of each bacterium was taken and was then divided into cases and control groups. Both groups were photosensitized using toluidine blue O (TBO dye and the case groups were irradiated with Ga-As laser. Bacterial colonies were then serially diluted and were incubated for subculture. After incubation period, the number of viable bacterial count was performed. Statistical Analysis: Wilcoxon-signed rank test was carried out to determine significance of reduction on subsequent dilution within the bacterial group. Mann–Whitney U-test was performed to determine the significance of reduction between cases and control of particular bacterial group. Results: The results revealed substantial reduction in the viable bacterial count. F. nucleatum was found to be most sensitive to killing by laser irradiation followed by P. intermedia and

Essential Oil Composition of the Different Parts and In Vitro Shoot Culture of Eryngium planum L.

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Anna Kurowska

2011-08-01

Full Text Available The essential oils obtained by hydrodistillation from the different parts (inflorescence, stalk leaves, rosette leaves and root as well as from in vitro shoot culture of Eryngium planum L. were analyzed by GC-FID-MS in respect to their chemical composition. The different parts of E. planum and in vitro shoots showed different yields. The part with higher amount was the inflorescences, followed by the stalk leaves and in vitro shoots, rosette leaves and finally roots. The essential oils obtained from rosette leaves and in vitro-derived rosettes had totally different composition. Quantitative differences were also found between compounds of intact plant organs. The main components of stalk leaf oil and rosette leaf oil were monoterpene (limonene, α- and β-pinene and sesquiterpene hydrocarbons. In inflorescence oil cis-chrysanthenyl acetate (43.2% was accompanied by other esters (propionate, butanoate, hexanoate and octanoate and numerous oxygenated sesquiterpenes. Root oil and in vitro shoot oil contained mainly (Z-falcarinol and 2,3,4-trimethylbenzaldehyde. This is the first report on the chemical composition of this species.

Cytotoxic Effects of Dillapiole on Embryonic Development of Mouse Blastocysts in Vitro and in Vivo

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Wen-Hsiung Chan

2014-06-01

Full Text Available We examined the cytotoxic effects of dillapiole, a phenylpropanoid with antileishmanial, anti-inflammatory, antifungal, and acaricidal activities, on the blastocyst stage of mouse embryos, subsequent embryonic attachment and outgrowth in vitro, and in vivo implantation via embryo transfer. Blastocysts treated with 2.5–10 μM dillapiole exhibited a significant increase in apoptosis and corresponding decrease in total cell number. Notably, the implantation success rates of blastocysts pretreated with dillapiole were lower than those of their control counterparts. Moreover, in vitro treatment with 2.5–10 μM dillapiole was associated with increased resorption of post-implantation embryos and decreased fetal weight. Our results collectively indicate that dillapiole induces apoptosis and retards early post-implantation development, both in vitro and in vivo. However, the extent to which this organic compound exerts teratogenic effects on early human development is not known at present. Further studies are required to establish effective protection strategies against the cytotoxic effects of dillapiole.

In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells.

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Fang, Mingzhu; Kang, Hwan-Goo; Park, Youngil; Estrella, Brian; Zarbl, Helmut

2017-09-28

The circadian rhythm is a fundamental physiological process present in all organisms that regulates biological processes ranging from gene expression to sleep behavior. In vertebrates, circadian rhythm is controlled by a molecular oscillator that functions in both the suprachiasmatic nucleus (SCN; central pacemaker) and individual cells comprising most peripheral tissues. More importantly, disruption of circadian rhythm by exposure to light-at-night, environmental stressors and/or toxicants is associated with increased risk of chronic diseases and aging. The ability to identify agents that can disrupt central and/or peripheral biological clocks, and agents that can prevent or mitigate the effects of circadian disruption, has significant implications for prevention of chronic diseases. Although rodent models can be used to identify exposures and agents that induce or prevent/mitigate circadian disruption, these experiments require large numbers of animals. In vivo studies also require significant resources and infrastructure, and require researchers to work all night. Thus, there is an urgent need for a cell-type appropriate in vitro system to screen for environmental circadian disruptors and enhancers in cell types from different organs and disease states. We constructed a vector that drives transcription of the destabilized luciferase in eukaryotic cells under the control of the human PERIOD 2 gene promoter. This circadian reporter construct was stably transfected into human mammary epithelial cells, and circadian responsive reporter cells were selected to develop the in vitro bioluminescence assay. Here, we present a detailed protocol to establish and validate the assay. We further provide details for proof of concept experiments demonstrating the ability of our in vitro assay to recapitulate the in vivo effects of various chemicals on the cellular biological clock. The results indicate that the assay can be adapted to a variety of cell types to screen for both

In vitro and in vivo toxicity assessment of nanoparticles

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Kumar, Vinay; Sharma, Neha; Maitra, S. S.

2017-11-01

Nanotechnology has revolutionized gene therapy, diagnostics and environmental remediation. Their bulk production, uses and disposal have posed threat to the environment. With the appearance of these nanoparticles in the environment, their toxicity assessment is an immediate concern. This review is an attempt to summarize the major techniques used in cytotoxity determination. The review also presents a detailed and elaborative discussion on the toxicity imposed by different types of nanoparticles including carbon nanotubes, gold nanoparticles, silver nanoparticles, quantum dots, fullerenes, aluminium nanoparticles, zinc nanoparticles, iron nanoparticles, titanium nanoparticles and silica nanoparticles. It discusses the in vitro and in vivo toxological effects of nanoparticles on bacteria, microalgae, zebrafish, crustacean, fish, rat, mouse, pig, guinea pig, human cell lines and human. It also discusses toxological effects on organs such as liver, kidney, spleen, sperm, neural tissues, liver lysosomes, spleen macrophages, glioblastoma cells, hematoma cells and various mammalian cell lines. It provides information about the effects of nanoparticles on the gene-expression, growth and reproduction of the organisms.

Effect of electron beam on in vitro cultured orchid organs

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Ryu, Jaihyunk; Bae, Seho; Bae, Changhyu [Sunchon National Univ., Suncheon (Korea, Republic of); Kang, Hyun Suk; Lee, Byung Cheol [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2010-07-01

Ionizing radiations have been effective mutagen sources to overcome the limitation of the useful genetic resources in natural environment. The study was conducted to investigate an effect of electron beam on organogenesis, growth patterns and genetic variation in the irradiated orchid organs. The in utero cultured rhizomes of orchids were irradiated with the electron beam in the dose range of 15Gy to 2240Gy under the condition of various beam energy and beam current. Significant decreases in survival, growth and organogenesis were observed by increase of intensity of electron beam irradiation. The irradiation intensity of lethal dose 50 of the in utero cultured orchid was estimated as approximately 500Gy to 1000Gy under 10MeV/n, and 1000Gy was optimal for growth and organogenesis of the cultures under 10MeV/n with 0.05mA treatment, and 15Gy {approx} 48Gy under 2MeV/n and 0.5mA electron beam condition. RAPD and ISSR analyses for the electron beam irradiated organs were performed to analyze genetic variation under the electron beam condition. Both of RAPD and ISSR analyses showed higher polymorphic rate in the electron-beam irradiated C. gangrene and C. Kaner.

In vitro gastrointestinal digestion increases the translocation of polystyrene nanoparticles in an in vitro intestinal co-culture model.

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Walczak, Agata P; Kramer, Evelien; Hendriksen, Peter J M; Helsdingen, Richard; van der Zande, Meike; Rietjens, Ivonne M C M; Bouwmeester, Hans

2015-01-01

The conditions of the gastrointestinal tract may change the physicochemical properties of nanoparticles (NPs) and therewith the bioavailability of orally taken NPs. Therefore, we assessed the impact of in vitro gastrointestinal digestion on the protein corona of polystyrene NPs (PS-NPs) and their subsequent translocation across an in vitro intestinal barrier. A co-culture of intestinal Caco-2 and HT29-MTX cells was exposed to 50 nm PS-NPs of different charges (positive and negative) in two forms: pristine and digested in an in vitro gastrointestinal digestion model. In vitro digestion significantly increased the translocation of all, except the "neutral", PS-NPs. Upon in vitro digestion, translocation was 4-fold higher for positively charged NPs and 80- and 1.7-fold higher for two types of negatively charged NPs. Digestion significantly reduced the amount of protein in the corona of three out of four types of NPs. This reduction of proteins was 4.8-fold for "neutral", 3.5-fold for positively charged and 1.8-fold for one type of negatively charged PS-NPs. In vitro digestion also affected the composition of the protein corona of PS-NPs by decreasing the presence of higher molecular weight proteins and shifting the protein content of the corona to low molecular weight proteins. These findings are the first to report that in vitro gastrointestinal digestion significantly affects the protein corona and significantly increases the in vitro translocation of differently charged PS-NPs. These findings stress the importance of including the in vitro digestion in future in vitro intestinal translocation screening studies for risk assessment of orally taken NPs.

Propagação in vitro de Sacha inchi In vitro propagation of Sacha inchi

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Stevan Ricardo Bordignon

2012-07-01

Full Text Available O objetivo do presente trabalho foi avaliar in vitro a relação auxina:citocinina a fim de obter propágulos nos segmentos distintos do epicótilo e hipocótilo de sementes germinadas in vitro de Sacha inchi (Plukenetia volubilis Linneo. Os segmentos apical (A, mediano (B e o basal (C foram introduzidos em meio de cultivo MS, semi sólido (2,0g L-1 Phytagel, suplementado com vitaminas de MS, sacarose (30,0g L-1 e submetidos a três doses da auxina ácido indolbutírico - IBA (0; 0,1; 0,5mg L-1, associadas a quatro doses da citocinina benzilaminopurina - BAP (0; 0,1; 0,5; 1,0mg L-1, totalizando 36 tratamentos. Após nove semanas de cultivo in vitro, o segmento apical (A apresentou formação de brotações por organogênese direta nas concentrações de 0,5 e 1,0 de BAP, associado a 0,0 e 0,1 de IBA. O emprego do cultivo in vitro é viável na produção de mudas, utilizando como explante a região apical de sementes germinadas in vitro.The aim of this study was to perform an in vitro evaluation of the auxin:cytokinine ratio in different segments of the epicotyl and hypocotyl of Sacha inchi (Plukenetia volubilis Linneo seeds germinated in vitro. The segments apical (A, median (B and basal (C were introduced into semi-solid MS culture medium (2.0g L-1 Phytagel, supplemented with MS vitamins, sucrose (30.0g L-1 and submitted to three doses of auxin indolebutyric acid - IBA (0; 0.1; 0.5mg L-1, associated with four doses of the cytokinine benzylaminopurine - BAP (0; 0.1; 0.5; 1.0mg L-1, totaling 36 treatments. After nine weeks of in vitro cultivation, the apical segment (A presented shoot formation by direct organogenesis at the concentrations of 0.5 and 1.0 of BAP associated with 0.0 and 0.1 of IBA. It is feasible to use in vitro cultivation with the apical region of seeds germinated in vitro used as explants.

Detecting of Mycoplasma genitalium in male patients with urethritis symptoms in Turkey by polymerase chain reaction

International Nuclear Information System (INIS)

Dolapci, Istar; Tekeli, Alper; Ozsan, Murat; Elhan, Atilla; Yaman, Onder; Ergin, Sureyya

2005-01-01

The aim of this study was to investigate the incidence of Mycoplasma genitalium in the urine samples of 63 male patients who had urethritis symptoms. Along with Neisseria gonorrhoeae (N. gonorrhoeae) and Chlamydia trachomatis (C. trachomatis). We also investigated Mycoplasma hominis (M. hominis) and Ureaplasma urealyticum (U. urealyticum), both of which are known to cause urethritis. Microorganisms were investigated in urine samples of the patients with polymerase chain reaction. The study was conducted between September 2003 - February 2004 at the Department of Microbiology and Clinical Microbiology Ankara University School of Medicine, Ankara, Turkey. A total of 63 urine samples were analyzed and 6 (9.52%) patients had N. gonorrhoeae, 4 (6.34%) had C. trachomatis, while 4 (6.34%) urines were positive in terms of M. genitalium. Nevertheless, 3 (4.76%) patients had U. urealyticum and 2 (3.17%) patients had M. hominis. One urine sample was positive in terms of both N. gonorrhoeae and U. urealyticum, and another urine sample was positive in terms of both M. hominis and U. urealyticum. The results were compared with the control group and found no statistically significant difference. Mycoplasma species are found in normal flora of urogenital system and also as an agent of urogenital infection. In our study, we found low microorganism rates when compared with Europe and America. This difference may be due to the conservative sexual behavior in Turkey. (author)

Nutrient Changes and in Vitro Digestibility in Generative Stage of M10-BMR Sorghum Mutant Lines

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R. Sriagtula

2017-08-01

Full Text Available The objective of this research was to investigate the influences of generative stage on crude protein, crude fiber, ash, and crude fat contents as well as in-vitro dry matter and organic matter digestibilities of M-10 BMR sorghum mutant lines. This research was arranged into a randomized block design with 2 factors. The first factor was M-10 BMR sorghum mutant lines (Patir 3.1, Patir 3.2 and Patir 3.7 and the second factor was generative stages (flowering, soft dough and hard dough phase. The observed variables were proximate contents of stem, leaves and panicle of sorghum plant and in-vitro digestibility of whole plant. The results showed that leaves crude protein (CP was more influenced by M-10 BMR sorghum mutant lines. Stems and panicles CP were influenced by the interaction between M-10 BMR sorghum mutant lines and generative stages. Further generative stage reduced stems CP but increased panicles CP. Crude fiber (CF, ash, and ether extract (EE in leaves were not influenced by generative stages. Stems CF was influenced by M-10 BMR sorghum mutant lines and generative stages, while stems EE was more influenced by generative stages. Stems ash content was influenced by the interaction between M-10 BMR sorghum mutant lines and generative stages while panicles ash content was more influenced by generative stages. M-10 BMR sorghum mutant lines and hard dough phase increased in-vitro dry matter and organic matter digestibilities. Based on those findings, it can be concluded that the increased maturity reduces CP and CF contents so it increases in-vitro digestibilities.

In vitro digestibility of lucerne hay using bag method

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constantin gavan

2013-10-01

Full Text Available A filter bag method was used for estimating apparent dry matter (DM digestibility, apparent digestible organic matter in DM, true DM digestibility, true digestible organic matter in DM and Digestibility of neutral detergent fibre. The forage sample analysed comprised tree plant parts (whole, leaf and stem of alfalfa (Medicago sativa, one degree of particle breakdown (1 mm sieve size at miliing and two field replicates at Agricultural Research and Developement Station (ARDS Șimnic-Craiova. Rumen fluid was used from two cannulated dairy cows. The use of filter bags can give acceptable results for the in vitro digestibility of forages. The escape of soluble matter from bags with samples high in solubles could influence the microbial population and hence increase cell wall degradation in bags with samples low in soluble matter, if  all are in the same incubation vessel.

Prevalence of Cryptosporidium spp. and other intestinal parasites in children with diarrhea

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Mutalip Çiçek

2011-03-01

Full Text Available This study was planned to determine the role of Cryptosporidium sp. and other intestinal parasites in the diarrheal diseases in children with 0-15 years old Van district.Materials and methods: In this study, stool samples of 450 children were examined for parasites. In the study, nativ-lugol, formaldehyde-ethyl acetate sedimentation methods and trichrome staining methods were used to detect parasites in stool samples. Additionally, sedimentation methods and modified acid fast staining method were used to detect the Cryptosporidium oocysts.Results: Parasites were found in 154 (34.2% among 450 children’s with diarrhea. In this study; the ratios of parasites were as follow: Giardia intestinalis 13.5%, Blastocystis hominis 10%, Entamoeba coli 3.78%, Cryptosporidium spp. 2.2%, Hymenolepis nana 1.33 %ve Ascaris lumbricoides 1.11%.Entamoeba histolytica/Entamoeba dispar 0.89%, Chilomastix mesnili 1.78%, Iodamoeba butschlii 0.89%, Entamoeba hartmanni 0.89%, Trichomonas hominis 0.67%, Enteromonas hominis 0.67%,Conclusion: In the investigate, it was found that Giardia intestinalis and Blastocystis hominis were most prominent agents in children with diarrhea in our vicinity and Cryptosporidium spp also was an important agent which should be investigated carefully in especially risk group in routine laboratory studies.

Prospective clinical testing of regulatory dendritic cells (DCreg) in organ transplantation

OpenAIRE

ANGUS W THOMSON; ALAN F ZAHORCHAK; Mohamed B. Ezzelarab; Lisa H. Butterfield; Fadi G. Lakkis; Diana M Metes

2016-01-01

Dendritic cells (DC) are rare, professional antigen-presenting cells with ability to induce or regulate alloimmune responses. Regulatory DC (DCreg) with potential to down-modulate acute and chronic inflammatory conditions that occur in organ transplantation can be generated in vitro under a variety of conditions. Here, we provide a rationale for evaluation of DCreg therapy in clinical organ transplantation with the goal of promoting sustained, donor-specific hyporesponsiveness, while lowering...

Dietary fibers from mushroom sclerotia: 3. In vitro fermentability using human fecal microflora.

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Wong, Ka-Hing; Wong, King-Yee; Kwan, Hoi-Shan; Cheung, Peter C K

2005-11-30

The in vitro fermentability of three novel dietary fibers (DFs) prepared from mushroom sclerotia, namely, Pleurotus tuber-regium, Polyporous rhinocerus, and Wolfiporia cocos, was investigated and compared with that of the cellulose control. All DF samples (0.5 g each) were fermented in vitro with a human fecal homogenate (10 mL) in a batch system (total volume, 50 mL) under strictly anaerobic conditions (using oxygen reducing enzyme and under argon atmosphere) at 37 degrees C for 24 h. All three novel sclerotial DFs exhibited notably higher dry matter disappearance (P. tuber-regium, 8.56%; P. rhinocerus, 13.5%; and W. cocos, 53.4%) and organic matter disappearance (P. tuber-regium, 9.82%; P. rhinocerus, 14.6%; and W. cocos, 57.4%) when compared with those of the cellulose control. Nevertheless, only the W. cocos DF was remarkably degraded to produce considerable amounts of total short chain fatty acids (SCFAs) (5.23 mmol/g DF on organic matter basis, with a relatively higher molar ratio of propionate) that lowered the pH of its nonfermented residue to a slightly acidic level (5.89). Variations on the in vitro fermentability among the three sclerotial DFs might mainly be attributed to their different amounts of interwoven hyphae present (different amounts of enzyme inaccessible cell wall components) as well as the possible different structural arrangement (linkage and degree of branching) of their beta-glucans.

Micro pore arrays in free standing cyclic olefin copolymer membranes: fabrication and surface functionalization strategies for in-vitro barrier tissue models

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Gel, M.; Kandasamy, S.; Cartledge, K.; Be, C. L.; Haylock, D.

2013-12-01

In recent years there has been growing interest in micro engineered in-vitro models of tissues and organs. These models are designed to mimic the in-vivo like physiological conditions with a goal to study human physiology in an organ-specific context or to develop in-vitro disease models. One of the challenges in the development of these models is the formation of barrier tissues in which the permeability is controlled locally by the tissues cultured at the interface. In-vitro models of barrier tissues are typically created by generating a monolayer of cells grown on thin porous membranes. This paper reports a robust preparation method for free standing porous cyclic olefin copolymer (COC) membranes. We also demonstrate that gelatin coated membranes facilitate formation of highly confluent monolayer of HUVECs. Membranes with thickness in the range of 2-3 um incorporating micro pores with diameter approximately 20 um were fabricated and integrated with microfluidic channels. The performance of the device was demonstrated with a model system mimicking the endothelial barrier in bone marrow sinusoids.

In vitro and in vivo study of the antibacterial effects of Nigella sativa methanol extract in dairy cow mastitis

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Hassan Rakhshandeh

2011-07-01

Results and conclusion: The extract showed significant in vitro and in vivo inhibitory effects on causative organisms compared to standard drugs and also induced healing of the disease. This is the first veterinary experiment, to our knowledge, that investigated the antibacterial effects of Nigella sativa.

In vitro studies of the physical interactions between neurofilaments, microtubules and mitochondria isolated from the central nervous system

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Leterrier, Jean-François; Eyer, Joël; Weiss, Dieter G.; Lindén, Monica

1991-05-01

In order to explore the molecular nature and the regulation of dense cytomatrix which interconnects MT, NF and membranous organelles in neurons (9), the interactions between NF, MT and each of these cytoskelatal elements with brain mitochondria were investigated in vitro using biochemical and viophysical methods. From these studies, the following conclusions were drawn: 1- Pure NF form in vitro a highly viscous gel, dependent upon the phosphorylation state of the side arms of the NF-H and M subunits which might participate directly to the interactions since antibodies specific of these phosphorylated sites inhibited efficiently the NF gelation. This process is modulated by both ATP hydrolysis and soluble molecules from nervous tissue and it might reflect the highly controled organization of NF bundles in axons. 2- In contrast with NF, low viscosity levels were detected in MT suspensions. However, the occurrence of weak interactions between MT were deduced from studies with taxol, ATP, AMP-PNP and Mg ions, which affected the viscosity and the organization of MT in vitro, possibly through MAPs mediated interactions. 3- Mitochondria associated permanently in vitro to few MT through cross-bridges involving MAPs, which bind to specific sites on the outer membrane (17). In addition, brain mitochondria (and not liver mitochondria) interact with NF in an ATP-dependent manner, through thin cross-bridges possibly involving the NF-H and M subunits since these molecules, when purified, compete efficiently with MAPs for the binding to membrane sites. These results suggest the participation of structure MAPs and of NF-H and M subunits in the spatial organization MT and NF and in anchoring mitochondria to the cytomatrix.

SYTO-13, a Viability Marker as a New Tool to Monitor In Vitro Pharmacodynamic Parameters of Anti-Pneumocystis Drugs.

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Annie Standaert-Vitse

Full Text Available While Pneumocystis pneumonia (PcP still impacts the AIDS patients, it has a growing importance in immunosuppressed HIV-negative patients. To determine the anti-Pneumocystis therapeutic efficacy of new compounds, animal and in vitro models have been developed. Indeed, well-designed mouse or rat experimental models of pneumocystosis can be used to describe the in vivo anti-Pneumocystis activity of new drugs. In vitro models, which enable the screening of a large panel of new molecules, have been developed using axenic cultures or co-culture with feeder cells; but no universally accepted standard method is currently available to evaluate anti-Pneumocystis molecules in vitro. Thus, we chose to explore the use of the SYTO-13 dye, as a new indicator of Pneumocystis viability. In the present work, we established the experimental conditions to define the in vitro pharmacodynamic parameters (EC50, Emax of marketed compounds (trimethoprim/sulfamethoxazole, pentamidine, atovaquone in order to specifically measure the intrinsic activity of these anti-P. carinii molecules using the SYTO-13 dye for the first time. Co-labelling the fungal organisms with anti-P. carinii specific antibodies enabled the measurement of viability of Pneumocystis organisms while excluding host debris from the analysis. Moreover, contrary to microscopic observation, large numbers of fungal cells can be analyzed by flow cytometry, thus increasing statistical significance and avoiding misreading during fastidious quantitation of stained organisms. In conclusion, the SYTO-13 dye allowed us to show a reproducible dose/effect relationship for the tested anti-Pneumocystis drugs.

In vitro and in vivo activity of an organic tellurium compound on Leishmania (Leishmania chagasi.

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Isabella Aparecida Salerno Pimentel

Full Text Available Tellurium compounds have shown several biological properties and recently the leishmanicidal effect of one organotellurane was demonstrated. These findings led us to test the effect of the organotellurium compound RF07 on Leishmania (Leishmania chagasi, the agent of visceral leishmaniasis in Latin America. In vitro assays were performed in L. (L. chagasi-infected bone marrow derived macrophages treated with different concentrations of RF07. In in vivo experiments Golden hamsters were infected with L. (L. chagasi and injected intraperitoneally with RF07 whereas control animals received either Glucantime or PBS. The effect of RF07 on cathepsin B activity of L. (L. chagasi amastigotes was assayed spectrofluorometrically using fluorogenic substrates. The main findings were: 1 RF07 showed significant leishmanicidal activity against intracellular parasites at submicromolar concentrations (IC50 of 529.7±26.5 nM, and the drug displayed 10-fold less toxicity to macrophages (CC50 of 5,426±272.8 nM; 2 kinetics assays showed an increasing leishmanicidal action of RF07 at longer periods of treatment; 3 one month after intraperitoneal injection of RF07 L. (L. chagasi-infected hamsters showed a reduction of 99.6% of parasite burden when compared to controls that received PBS; 4 RF07 inhibited the cathepsin B activity of L. (L. chagasi amastigotes. The present results demonstrated that the tellurium compound RF07 is able to destroy L. (L. chagasi in vitro and in vivo at concentrations that are non toxic to the host. We believe these findings support further study of the potential of RF07 as a possible alternative for the chemotherapy of visceral leishmaniasis.

Physiology of in vitro culture

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Maria Jesús Cañal

2001-01-01

Full Text Available The culture procedures described up to the eighties, did not made any mention to the environmental control of in vitro plant development. However, growth rate, development and many of the physiologic-morphologic features of the in vitro grown plants are influenced by the culture vessel. The increasing knowledge about the environmental control of culture vessels under sterile conditions, is helping to change micorpropagation procedures. The in vitro environment with lower rate ventilation, brings about low flow rates of matter and energy, with minimum variations of temperature, high relative humidity and large daily changes of the concentration of CO2 inside the culture vessel. The type of culture vessel (size, shape, fabric and closing system can influence the evolution of the atmosphere along the time of culture. Although submitted to different stresses factors plant can be grown in vitro, but plants can be faulty in their anatomy, morphology and physiology. As a consequence, these plants shown a phenotype unable to survive to ex vitro conditions. Different strategies can be used to control the atmosphere along the different phases of micropropagation, in heterotrophic, mixotrophic or autotrophic cultures. The election of the best strategy will be based on different factors as species, number of transplantes required, or quality-price relationship. enviromental control, tissue culture, micropropagation Keywords: in vitro enviromental, characteristic physiology,

Effect of Acrylamide on Oocyte Nuclear Maturation and Cumulus Cells Apoptosis in Mouse In Vitro.

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Shuzhen Liu

Full Text Available Acrylamide (ACR is a chemical compound with severe neurotoxicity, genotoxicity, carcinogenicity and reproductive toxicity. Recent studies showed that ACR impairs the function of reproductive organs, e.g., epididymis and testes. In vitro maturation of mouse oocyte is a sensitive assay to identify potential chemical hazard to female fertility. The aim of this study was to evaluate the adverse effects of ACR on the nuclear maturation and cumulus cells apoptosis of mouse oocytes in vitro. Cumulus-oocyte complexes were incubated in a maturation medium containing 0, 5, 10 and 20 μM of ACR. Chromosome alignment and spindle morphology of oocytes was determined by immunofluorescence and confocal microscopy. Our results showed that oocytes exposed to different doses of ACR in vitro were associated with a significant decrease of oocyte maturation, significant increase of chromosome misalignment rate, occurrence of abnormal spindle configurations, and the inhibition of oocyte parthenogenetic activation. Furthermore, apoptosis of cumulus cells was determined by TUNEL and CASPASE-3 assay. Results showed that apoptosis in cumulus cells was enhanced and the expression of CASPASE-3 was increased after cumulus-oocyte complexes were exposed to ACR. Therefore, ACR may affect the nuclear maturation of oocytes via the apoptosis of cumulus cells in vitro.

Regeneration of Pelargonium in vitro

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Agnieszka Wojtania

2013-12-01

Full Text Available Pelargonium sp. has been a subject of numerous studies to deterimine the effec tiveness of in vitro techniques to produce a large number of pathogen-free plants. Regeneration of pelargonium plants from the different initial explants as well via organogenesis as via somatic embryogenesis has been obtained. The most effective adventitious shoot formation has been achieved from shoot tips and axillary buds using cytokinin or cytokinin/auxin combinations. Leaf explants, whose general have lower organogenic potency, regenerate better in the presence of thidiazuron. This growth regulator stimulate the somatic embryos production from hypocotyl and cotyledone explants too. The main problem in tissue culture propagation of Pelargonium has been the high tendency to formation of vigorously growing callus with low organogenic potency and rapid senescence of cultures. Moreover, the significant differen ces in requirements to the medium composition (minerals, organic compounds and growth regulators between Pelargonium cultivars has been observed. This makes difficult to develop an universaI method of Pelargonium micropropagation.

Proximate analysis, in vitro organic matter digestibility, and energy content of common guava (Psidium guajava L.) and yellow, strawberry guava (Psidium cattleianum Var. lucidum) tree parts and fruits as potential forage.

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Adrian, Julie Ann Luiz; Arancon, Norman Q; Mathews, Bruce W; Carpenter, James R

2012-10-24

The nutrient composition of common guava, Psidium guajava L., and strawberry guava (waiwi), Psidium cattleianum var. lucidum, tree parts and fruits was determined during three seasons for six locations in Hawaii to assess guava as a potential feed for cattle. All guava plant parts were higher (p Guava leaves were higher in fiber and had lower energy densities (p Guava fruits were higher in CP (p guava is low in vitro organic matter digestibility as compared to tropical forage grasses; therefore, it is not recommended as a feedstock for livestock.

Ferrocenyl and organic novobiocin derivatives: Synthesis and their in vitro biological activity.

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Mbaba, Mziyanda; Mabhula, Amanda N; Boel, Natasha; Edkins, Adrienne L; Isaacs, Michelle; Hoppe, Heinrich C; Khanye, Setshaba D

2017-07-01

A focused series of novobiocin derivatives containing a ferrocene unit together with their corresponding organic novobiocin analogues have been synthesized in modest to good yields. These compounds were screened for biological activity against a chloroquine-sensitive strain of Plasmodium falciparum (3D7) and human breast cancer cell line (HCC38). With the exception of compounds 5c and 5d, the general trend observed is that incorporation of the ferrocene moiety into novobiocin scaffold resulted in compounds 6a-d/6f showing enhanced activity compared to organic analogues 5a-b and 5e-f. Copyright © 2017 Elsevier Inc. All rights reserved.

The changes of nutrient composition and in vitro evaluation on gamma irradiated sweet sorghum bagasse

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Teguh Wahyono; Firsoni

2016-01-01

In vitro rumen fermentation study was done to evaluate the effects of gamma irradiation on nutrient compound changes and rumen fermentation product of sweet sorghum bagasse (SSB). The level doses 0, 50, 100 and 150 kGy from cobalt-60 gamma rays irradiator was used to treate sweet sorghum bagasse (SSB). Variables measured were nutrient values, gas production, methane (CH_4) production, total volatile fatty acid (TVFA), ammonia (NH_3), in vitro dry matter digestibility (IVDMD) and in vitro organic matter digestibility (IVOMD) after 72 h in-vitro incubation times. Complete randomized design (CRD) (four treatments and four replications) was used to analyze data. The results showed that gamma irradiation doses of 50, 100 and 150 kGy were able to reduce neutral detergent fibre (NDF) (2.15; 3.29 and 5.44% respectively) and acid detergent fibre (ADF) (3.29; 4.58 and 4.58% respectively) and significantly different (P <0.05). Gamma irradiation was capable to increase total volatile fatty acid (TVFA), IVDMD and IVOMD (P <0.05). Irradiation doses of 100 and 150 kGy also increased protozoa population and CH_4 production significantly (P <0.05). Gamma irradiation improved in vitro rumen performance represented in rumen fermentation products. (author)

Quantum dot labeling and tracking of cultured limbal epithelial cell transplants in-vitro

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Genicio, Nuria; Paramo, Juan Gallo; Shortt, Alex J.

2015-01-01

PURPOSE Cultured human limbal epithelial cells (HLEC) have shown promise in the treatment of limbal stem cell deficiency but little is known about their survival, behaviour and long-term fate post transplantation. The aim of this research was to evaluate, in-vitro, quantum dot (QDot) technology as a tool for tracking transplanted HLEC. METHODS In-vitro cultured HLEC were labeled with Qdot nanocrystals. Toxicity was assessed using live-dead assays. The effect on HLEC function was assessed using colony forming efficiency assays and expression of CK3, P63alpha and ABCG2. Sheets of cultured HLEC labeled with Qdot nanocrystals were transplanted onto decellularised human corneo-scleral rims in an organ culture model and observed to investigate the behaviour of transplanted cells. RESULTS Qdot labeling had no detrimental effect on HLEC viability or function in-vitro. Proliferation resulted in a gradual reduction in Qdot signal but sufficient signal was present to allow tracking of cells through multiple generations. Cells labeled with Qdots could be reliably detected and observed using confocal microscopy for at least 2 weeks post transplantation in our organ culture model. In addition it was possible to label and observe epithelial cells in intact human corneas using the Rostock corneal module adapted for use with the Heidelberg HRA. CONCLUSIONS This work demonstrates that Qdots combined with existing clinical equipment could be used to track HLEC for up to 2 weeks post transplantation, however, our model does not permit the assessment of cell labeling beyond 2 weeks. Further characterisation in in-vivo models are required. PMID:26024089

In vitro eugenics.

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Sparrow, Robert

2014-11-01

A series of recent scientific results suggest that, in the not-too-distant future, it will be possible to create viable human gametes from human stem cells. This paper discusses the potential of this technology to make possible what I call 'in vitro eugenics': the deliberate breeding of human beings in vitro by fusing sperm and egg derived from different stem-cell lines to create an embryo and then deriving new gametes from stem cells derived from that embryo. Repeated iterations of this process would allow scientists to proceed through multiple human generations in the laboratory. In vitro eugenics might be used to study the heredity of genetic disorders and to produce cell lines of a desired character for medical applications. More controversially, it might also function as a powerful technology of 'human enhancement' by allowing researchers to use all the techniques of selective breeding to produce individuals with a desired genotype. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Spatial organization of mesenchymal stem cells in vitro--results from a new individual cell-based model with podia.

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Martin Hoffmann

Full Text Available Therapeutic application of mesenchymal stem cells (MSC requires their extensive in vitro expansion. MSC in culture typically grow to confluence within a few weeks. They show spindle-shaped fibroblastoid morphology and align to each other in characteristic spatial patterns at high cell density. We present an individual cell-based model (IBM that is able to quantitatively describe the spatio-temporal organization of MSC in culture. Our model substantially improves on previous models by explicitly representing cell podia and their dynamics. It employs podia-generated forces for cell movement and adjusts cell behavior in response to cell density. At the same time, it is simple enough to simulate thousands of cells with reasonable computational effort. Experimental sheep MSC cultures were monitored under standard conditions. Automated image analysis was used to determine the location and orientation of individual cells. Our simulations quantitatively reproduced the observed growth dynamics and cell-cell alignment assuming cell density-dependent proliferation, migration, and morphology. In addition to cell growth on plain substrates our model captured cell alignment on micro-structured surfaces. We propose a specific surface micro-structure that according to our simulations can substantially enlarge cell culture harvest. The 'tool box' of cell migratory behavior newly introduced in this study significantly enhances the bandwidth of IBM. Our approach is capable of accommodating individual cell behavior and collective cell dynamics of a variety of cell types and tissues in computational systems biology.

In vitro-in vivo extrapolation: estimation of human serum concentrations of chemicals equivalent to cytotoxic concentrations in vitro

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Guelden, Michael; Seibert, Hasso

2003-01-01

In the present study an extrapolation model for estimating serum concentrations of chemicals equivalent to in vitro effective concentrations is developed and applied to median cytotoxic concentrations (EC 50 ) determined in vitro. Nominal concentrations of a chemical in serum and in vitro are regarded as equivalent, if they result in the same aqueous concentration of the unbound form. The algorithm used is based on equilibrium distribution and requires albumin binding data, the octanol-water partition coefficient (K ow ), and the albumin concentrations and lipid volume fractions in vitro and in serum. The chemicals studied cover wide ranges of cytotoxic potency (EC 50 : 2.5-530000 μM) and lipophilicity (log K ow : -5 to 7). Their albumin binding characteristics have been determined by means of an in vitro cytotoxicity test as described previously. The equivalent serum concentrations of 19 of the 33 compounds investigated, having high protein binding and/or lipophilicity, were substantially higher than the EC 50 -values, by factors of 2.5-58. Prominent deviations between the equivalent nominal concentrations in serum and in vitro were largely restricted to chemicals with higher cytotoxic potency (EC 50 ≤1000 μM). The results suggest that estimates of equivalent serum concentrations based on in vitro data are robust for chemicals with low lipophilicity (log K ow ≤2) and low potency (EC 50 >1000 μM). With more potent chemicals or those with higher lipophilicity partitioning into lipids and/or binding to serum proteins have to be taken into account when estimating in vivo serum concentrations equivalent to in vitro effective concentrations

Three-Dimensional In Vitro Skin and Skin Cancer Models Based on Human Fibroblast-Derived Matrix.

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Berning, Manuel; Prätzel-Wunder, Silke; Bickenbach, Jackie R; Boukamp, Petra

2015-09-01

Three-dimensional in vitro skin and skin cancer models help to dissect epidermal-dermal and tumor-stroma interactions. In the model presented here, normal human dermal fibroblasts isolated from adult skin self-assembled into dermal equivalents with their specific fibroblast-derived matrix (fdmDE) over 4 weeks. The fdmDE represented a complex human extracellular matrix that was stabilized by its own heterogeneous collagen fiber meshwork, largely resembling a human dermal in vivo architecture. Complemented with normal human epidermal keratinocytes, the skin equivalent (fdmSE) thereof favored the establishment of a well-stratified and differentiated epidermis and importantly allowed epidermal regeneration in vitro for at least 24 weeks. Moreover, the fdmDE could be used to study the features of cutaneous skin cancer. Complementing fdmDE with HaCaT cells in different stages of malignancy or tumor-derived cutaneous squamous cell carcinoma cell lines, the resulting skin cancer equivalents (fdmSCEs) recapitulated the respective degree of tumorigenicity. In addition, the fdmSCE invasion phenotypes correlated with their individual degree of tissue organization, disturbance in basement membrane organization, and presence of matrix metalloproteinases. Together, fdmDE-based models are well suited for long-term regeneration of normal human epidermis and, as they recapitulate tumor-specific growth, differentiation, and invasion profiles of cutaneous skin cancer cells, also provide an excellent human in vitro skin cancer model.

Modularity in developmental biology and artificial organs: a missing concept in tissue engineering.

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Lenas, Petros; Luyten, Frank P; Doblare, Manuel; Nicodemou-Lena, Eleni; Lanzara, Andreina Elena

2011-06-01

Tissue engineering is reviving itself, adopting the concept of biomimetics of in vivo tissue development. A basic concept of developmental biology is the modularity of the tissue architecture according to which intermediates in tissue development constitute semiautonomous entities. Both engineering and nature have chosen the modular architecture to optimize the product or organism development and evolution. Bioartificial tissues do not have a modular architecture. On the contrary, artificial organs of modular architecture have been already developed in the field of artificial organs. Therefore the conceptual support of tissue engineering by the field of artificial organs becomes critical in its new endeavor of recapitulating in vitro the in vivo tissue development. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

An in vitro approach for comparative interspecies metabolism of agrochemicals.

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Whalley, Paul M; Bartels, Michael; Bentley, Karin S; Corvaro, Marco; Funk, Dorothee; Himmelstein, Matthew W; Neumann, Birgit; Strupp, Christian; Zhang, Fagen; Mehta, Jyotigna

2017-08-01

The metabolism and elimination of a xenobiotic has a direct bearing on its potential to cause toxicity in an organism. The confidence with which data from safety studies can be extrapolated to humans depends, among other factors, upon knowing whether humans are systemically exposed to the same chemical entities (i.e. a parent compound and its metabolites) as the laboratory animals used to study toxicity. Ideally, to understand a metabolite in terms of safety, both the chemical structure and the systemic exposure would need to be determined. However, as systemic exposure data (i.e. blood concentration/time data of test material or metabolites) in humans will not be available for agrochemicals, an in vitro approach must be taken. This paper outlines an in vitro experimental approach for evaluating interspecies metabolic comparisons between humans and animal species used in safety studies. The aim is to ensure, where possible, that all potential human metabolites are also present in the species used in the safety studies. If a metabolite is only observed in human in vitro samples and is not present in a metabolic pathway defined in the toxicological species already, the toxicological relevance of this metabolite must be evaluated. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Functional properties of hepatocytes in vitro are correlated with cell polarity maintenance.

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Zeigerer, Anja; Wuttke, Anne; Marsico, Giovanni; Seifert, Sarah; Kalaidzidis, Yannis; Zerial, Marino

2017-01-01

Exploring the cell biology of hepatocytes in vitro could be a powerful strategy to dissect the molecular mechanisms underlying the structure and function of the liver in vivo. However, this approach relies on appropriate in vitro cell culture systems that can recapitulate the cell biological and metabolic features of the hepatocytes in the liver whilst being accessible to experimental manipulations. Here, we adapted protocols for high-resolution fluorescence microscopy and quantitative image analysis to compare two primary hepatocyte culture systems, monolayer and collagen sandwich, with respect to the distribution of two distinct populations of early endosomes (APPL1 and EEA1-positive), endocytic capacity, metabolic and signaling activities. In addition to the re-acquisition of hepatocellular polarity, primary hepatocytes grown in collagen sandwich but not in monolayer culture recapitulated the apico-basal distribution of EEA1 endosomes observed in liver tissue. We found that such distribution correlated with the organization of the actin cytoskeleton in vitro and, surprisingly, was dependent on the nutritional state in vivo. Hepatocytes in collagen sandwich also exhibited faster kinetics of low-density lipoprotein (LDL) and epidermal growth factor (EGF) internalization, showed improved insulin sensitivity and preserved their ability for glucose production, compared to hepatocytes in monolayer cultures. Although no in vitro culture system can reproduce the exquisite structural features of liver tissue, our data nevertheless highlight the ability of the collagen sandwich system to recapitulate key structural and functional properties of the hepatocytes in the liver and, therefore, support the usage of this system to study aspects of hepatocellular biology in vitro. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.