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Sample records for single stool sample

  1. Folic acid absorption determined by a single stool sample test--a double-isotope technique. The folic acid absorption capacity in children.

    Science.gov (United States)

    Hjelt, K

    1989-10-01

    The fractional folic acid absorption (FAFol) was determined in 66 patients with various gastrointestinal diseases by a double-isotope technique, employing a single stool sample test (SSST) as well as a complete stool collection. The age of the patients ranged from 2.5 months to 16.8 years (mean 6.3 years). The test dose was administered orally and consisted of 50 micrograms of [3H]folic acid (monoglutamate) (approximately 20 muCi), carmine powder, and 2 mg 51CrCl3 (approximately 1.25 muCi) as the unabsorbable tracer. The whole-body radiation given to a 1-year-old child averaged 4.8 mrad only. The stool and napkin contents were collected and homogenized by the addition of 300 ml chromium sulfuric acid. A 300-ml sample of the homogenized stool and napkin contents, as well as 300 ml chromium sulfuric acid (75% vol/vol) containing the standards, were counted for the content of 51Cr in a broad-based well counter. The quantity of [3H]folic acid was determined by liquid scintillation, after duplicate distillation. Estimated by SSST, the FAFol, which employs the stool with the highest content of 51Cr corresponding to the most carmine-colored stool, correlated closely with the FAFol based on complete stool collection (r = 0.96, n = 39, p less than 0.0001). The reproducibility of FAFol determined by SSST was assessed from repeated tests in 18 patients. For a mean of 81%, the SD was 4.6%, which corresponded to a coefficient of variation of 5.7%.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. An improved stool concentration procedure for the detection of Cryptosporidium oocysts in Orang Asli stool samples.

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    Salleh, Fatmah Md; Moktar, Norhayati; Yasin, Azlin Mohd; Al-Mekhlafi, Hesham M; Anuar, Tengku Shahrul

    2014-11-01

    To improve the stool concentration procedure, we modified different steps of the standard formalin-ether concentration technique and evaluated these modifications by examining stool samples collected in the field. Seven samples were found positive by the modified formalin-ether concentration technique (M-FECT). Therefore, the M-FECT procedure provides enhanced detection of Cryptosporidium oocysts. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Plesiomonas shigelloides in stool samples of patients in the Venda ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-16

    Nov 16, 2009 ... This study determined the haemolytic, haemagglutinating and antibiotic susceptibility activities of. Plesiomonas shigelloides isolated from stool samples of patients attending different health centers in the Venda region of South Africa. P.shigelloides was isolated and identified using the API 20E, API.

  4. Clinical illnesses associated with isolation of dysgonic fermenter 3 from stool samples.

    OpenAIRE

    Blum, R N; Berry, C D; Phillips, M G; Hamilos, D L; Koneman, E W

    1992-01-01

    The clinical significance of the fastidious organism DF-3 isolated from stool cultures is unclear. We sought to improve our understanding of this organism and to further define its association with human disease. Stool cultures for DF-3 were obtained from three sources: an ongoing study of enteric pathogens in patients infected with the human immunodeficiency virus, a screening procedure in which all stool samples submitted for Clostridium difficile toxin assay were cultured for DF-3, and sto...

  5. Stool sample storage conditions for the preservation of Giardia intestinalis DNA

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    Salih Kuk

    2012-12-01

    Full Text Available Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT, +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

  6. Comparison of Individual and Pooled Stool Samples for the Assessment of Soil-Transmitted Helminth Infection Intensity and Drug Efficacy

    Science.gov (United States)

    Mekonnen, Zeleke; Meka, Selima; Ayana, Mio; Bogers, Johannes; Vercruysse, Jozef; Levecke, Bruno

    2013-01-01

    Background In veterinary parasitology samples are often pooled for a rapid assessment of infection intensity and drug efficacy. Currently, studies evaluating this strategy in large-scale drug administration programs to control human soil-transmitted helminths (STHs; Ascaris lumbricoides, Trichuris trichiura, and hookworm), are absent. Therefore, we developed and evaluated a pooling strategy to assess intensity of STH infections and drug efficacy. Methods/Principal Findings Stool samples from 840 children attending 14 primary schools in Jimma, Ethiopia were pooled (pool sizes of 10, 20, and 60) to evaluate the infection intensity of STHs. In addition, the efficacy of a single dose of mebendazole (500 mg) in terms of fecal egg count reduction (FECR; synonym of egg reduction rate) was evaluated in 600 children from two of these schools. Individual and pooled samples were examined with the McMaster egg counting method. For each of the three STHs, we found a significant positive correlation between mean fecal egg counts (FECs) of individual stool samples and FEC of pooled stool samples, ranging from 0.62 to 0.98. Only for A. lumbricoides was any significant difference in mean FEC of the individual and pooled samples found. For this STH species, pools of 60 samples resulted in significantly higher FECs. FECR for the different number of samples pooled was comparable in all pool sizes, except for hookworm. For this parasite, pools of 10 and 60 samples provided significantly higher FECR results. Conclusion/Significance This study highlights that pooling stool samples holds promise as a strategy for rapidly assessing infection intensity and efficacy of administered drugs in programs to control human STHs. However, further research is required to determine when and how pooling of stool samples can be cost-effectively applied along a control program, and to verify whether this approach is also applicable to other NTDs. PMID:23696905

  7. Detection of Gastrointestinal Pathogens from Stool Samples on Hemoccult Cards by Multiplex PCR

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    Martin Alberer

    2017-01-01

    Full Text Available Purpose. Up to 30% of international travelers are affected by travelers’ diarrhea (TD. Reliable data on the etiology of TD is lacking. Sufficient laboratory capacity at travel destinations is often unavailable and transporting conventional stool samples to the home country is inconvenient. We evaluated the use of Hemoccult cards for stool sampling combined with a multiplex PCR for the detection of model viral, bacterial, and protozoal TD pathogens. Methods. Following the creation of serial dilutions for each model pathogen, last positive dilution steps (LPDs and thereof calculated last positive sample concentrations (LPCs were compared between conventional stool samples and card samples. Furthermore, card samples were tested after a prolonged time interval simulating storage during a travel duration of up to 6 weeks. Results. The LPDs/LPCs were comparable to testing of conventional stool samples. After storage on Hemoccult cards, the recovery rate was 97.6% for C. jejuni, 100% for E. histolytica, 97.6% for norovirus GI, and 100% for GII. Detection of expected pathogens was possible at weekly intervals up to 42 days. Conclusion. Stool samples on Hemoccult cards stored at room temperature can be used in combination with a multiplex PCR as a reliable tool for testing of TD pathogens.

  8. DNA extraction in Echinococcus granulosus and Taenia spp. eggs in dogs stool samples applying thermal shock.

    Science.gov (United States)

    Hidalgo, Alejandro; Melo, Angélica; Romero, Fernando; Hidalgo, Víctor; Villanueva, José; Fonseca-Salamanca, Flery

    2018-03-01

    The extraction of DNA in taeniid eggs shows complications attached to the composition of stool samples and the high resistance of eggs to degradation. The objective of this study was to test a method of DNA extraction in taeniid eggs by applying a thermal shock to facilitate the chemical-enzymatic degradation of these elements. A group of six tubes containing 1 ml of dog stool sample was spiked with eggs of Echinococcus granulosus and another group of six with Taenia pisiformis. Samples were floated with supersaturated sugar solution and centrifuged. The upper portion of each tube (500 μl) was aspirated and deposited in 1.5 ml tubes. Three tubes from each group were incubated at -20 °C and then at 90 °C, the remaining three from each group, incubated at room temperature. Proteinase K and lysis buffer were added to each tube and incubated for 12 h at 58 °C. The lysis effect was evaluated by microscopy at 3, 6 and 12 h and integrity by electrophoresis in 1% agarose gels. With the same experimental scheme, the thermal shock effect was evaluated in extractions of 1, 2, 3 and 4 eggs of each species and the DNA was quantified. Additionally, the protocol was applied in samples of 4 dogs diagnosed with natural infection by Taeniidae worms. Finally, all the extractions were tested by PCR amplification. Both E. granulosus and T. pisiformis eggs showed a similar response in the tests. In samples without treatment, the lysis effect was poor and showed no differences over time, but in those subjected to thermal shock, eggs degradation increased with time. In both treatments, there was no DNA loss integrity. The protocol applied to limited amounts of eggs yielded PCR products in 100% of the samples exposed to thermal shock, allowing PCR amplifications up to 1 egg. In non-exposed samples, the results were not replicable. However, DNA quantification showed low values in both treatments. In turn, DNA extractions with thermal shock in infected dog samples

  9. Detection of Salmonella typhi by nested polymerase chain reaction in blood, urine, and stool samples

    NARCIS (Netherlands)

    Hatta, Mochammad; Smits, Henk L.

    2007-01-01

    A nested polymerase chain reaction (PCR) specific for Salmonella enterica serovar Typhi was used for the detection of the pathogen in blood, urine, and stool samples from 131 patients with clinical suspicion of typhoid fever. The sensitivity of blood culture, the PCRs with blood, urine, and feces,

  10. Stool DNA Test

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    ... The stool DNA test is a noninvasive laboratory test that identifies DNA changes in the cells of a stool sample. ... the presence of cancer. If a stool DNA test detects abnormal DNA, additional testing may be used to investigate the ...

  11. Clinical illnesses associated with isolation of dysgonic fermenter 3 from stool samples.

    Science.gov (United States)

    Blum, R N; Berry, C D; Phillips, M G; Hamilos, D L; Koneman, E W

    1992-02-01

    The clinical significance of the fastidious organism DF-3 isolated from stool cultures is unclear. We sought to improve our understanding of this organism and to further define its association with human disease. Stool cultures for DF-3 were obtained from three sources: an ongoing study of enteric pathogens in patients infected with the human immunodeficiency virus, a screening procedure in which all stool samples submitted for Clostridium difficile toxin assay were cultured for DF-3, and stool samples submitted specifically for DF-3 culture. Retrospective clinical data were obtained from chart reviews of patients with positive cultures. Antimicrobial susceptibility testing and cell wall fatty acid analysis were performed for each DF-3 isolated. Eight isolates of DF-3 were obtained over a period of 8 months. All patients either had severe underlying disease or were immunocompromised, including three patients coinfected with human immunodeficiency virus and two patients with inflammatory bowel disease. The spectrum of clinical disease ranged from chronic diarrhea with a well-defined response to therapy for DF-3 to an asymptomatic carrier state. Cell wall fatty acid analysis of these isolates demonstrated a consistent pattern with a large peak of 12-methyltetradecanoate. DF-3, a fastidious gram-negative coccobacillus, can be recovered from stool cultures of immunocompromised patients by using selective media. The presence of 12-methyltetradecanoate in cell wall fatty acid analysis assists in identification. The increased use of a selective medium-(cefoperazone-vancomycin-amphotericin B) in the evaluation of diarrhea in immunocompromised hosts, including persons with inflammatory bowel disease, may better define the association of DF-3 with human gastrointestinal disease.

  12. Acid fast cysts in diarrheal stool samples of HIV positive patients

    OpenAIRE

    Anuradha Mokkapati

    2015-01-01

    Background: Gastrointestinal infections are very common in patients with HIV infection or AIDS, and diarrhea is a common clinical presentation of these infections. Acid fast protozoans are very commonly responsible for diarrhea in HIV positive patients leading to death in many cases. Methods: The study group included 50 HIV seropositive patients suffering from diarrhea and the control group included 50 HIV seronegative patients suffering from diarrhea. The stool samples collected were con...

  13. Parasitological stool sample exam by spontaneous sedimentation method using conical tubes: effectiveness, practice, and biosafety

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    Steveen Rios Ribeiro

    2012-06-01

    Full Text Available INTRODUCTION: Spontaneous sedimentation is an important procedure for stool examination. A modification of this technique using conical tubes was performed and evaluated. METHODS: Fifty fecal samples were processed in sedimentation glass and in polypropylene conical tubes. Another 50 samples were used for quantitative evaluation of protozoan cysts. RESULTS: Although no significant differences occurred in the frequency of protozoa and helminths detected, significant differences in protozoan cyst counts did occur. CONCLUSIONS: The use of tube predicts a shorter path in the sedimentation of the sample, increases concentration of parasites for microscopy analysis, minimizes the risks of contamination, reduces the odor, and optimizes the workspace.

  14. Within-Stool and Within-Day Sample Variability of Fecal Calprotectin in Patients With Inflammatory Bowel Disease: A Prospective Observational Study.

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    Du, Lillian; Foshaug, Rae; Huang, Vivian W; Kroeker, Karen I; Dieleman, Levinus A; Halloran, Brendan P; Wong, Karen; Fedorak, Richard N

    2018-03-01

    The use of fecal calprotectin (FC) as a stool biomarker for differentiating inflammatory bowel disease (IBD) from IBS has been well validated, and there is a strong correlation between FC and the presence of endoscopic inflammatory lesions. However, recent studies have demonstrated intraindividual sample variability in patients with IBD, possibly limiting the reliability of using a single sample for monitoring disease activity. Our aim was to assess the within-stool and within-day sample variability of FC concentrations in patients with IBD. We examined a cross-sectional cohort of 50 adult IBD patients. Eligible patients were instructed to collect 3 samples from different parts of the stool from their first bowel movement of the day and 3 samples from each of up to 2 additional bowel movements within 24 hours. FC concentrations were measured by a rapid, quantitative point-of-care test using lateral flow technology (Quantum Blue). Descriptive statistics were used to assess FC variability within a single bowel movement and between different movements at different FC positivity cutoffs. Within a single bowel movement, there was clinically significant sample variability ranging from 8% to 23% depending on the time of the day or on the FC positivity cutoff value. Between bowel movements, there was clinically significant sample variability ranging from 13% to 26% depending on the FC positivity cutoff. Considering a single FC sample, the first sample of the day with an FC positivity cutoff of 250 μg/g provided the most reliable indication of disease activity.

  15. Evaluation of a Single Procedure Allowing the Isolation of Enteropathogenic Yersinia along with Other Bacterial Enteropathogens from Human Stools

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    Savin, Cyril; Leclercq, Alexandre; Carniel, Elisabeth

    2012-01-01

    Enteropathogenic Yersinia are among the most frequent agents of human diarrhea in temperate and cold countries. However, the incidence of yersiniosis is largely underestimated because of the peculiar growth characteristics of pathogenic Yersinia, which make their isolation from poly-contaminated samples difficult. The use of specific procedures for Yersinia isolation is required, but is expensive and time consuming, and therefore is not systematically performed in clinical pathology laboratories. A means to circumvent this problem would be to use a single procedure for the isolation of all bacterial enteropathogens. Since the Statens Serum Institut enteric medium (SSI) has been reported to allow the growth at 37°C of most Gram-negative bacteria, including Yersinia, our study aimed at evaluating its performances for Yersinia isolation, as compared to the commonly used Yersinia-specific semi-selective Cefsulodin-Irgasan-Novobiocin medium (CIN) incubated at 28°C. Our results show that Yersinia pseudotuberculosis growth was strongly inhibited on SSI at 37°C, and therefore that this medium is not suitable for the isolation of this species. All Yersinia enterocolitica strains tested grew on SSI, while some non-pathogenic Yersinia species were inhibited. The morphology of Y. enterocolitica colonies on SSI allowed their differentiation from various other Gram-negative bacteria commonly isolated from stool samples. However, in artificially contaminated human stools, the recovery of Y. enterocolitica colonies on SSI at 37°C was difficult and was 3 logs less sensitive than on CIN at 28°C. Therefore, despite its limitations, the use of a specific procedure (CIN incubated at 28°C) is still required for an efficient isolation of enteropathogenic Yersinia from stools. PMID:22911756

  16. Chromatography paper strip sampling of enteric adenoviruses type 40 and 41 positive stool specimens

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    Rahman Mustafizur

    2005-02-01

    Full Text Available Abstract Background The enteric subgroup F adenoviruses type 40 (Ad40 and 41 (Ad41 are the second most important cause of acute infantile gastroenteritis after rotaviruses. Repeated community outbreaks have been associated with antigenic changes among the Ad40 and Ad41 strains due to host immune pressure. Therefore large field epidemiological surveys and studies on the genetic variations in different isolates of Ad40 and Ad41 are important for disease control programs, the design of efficient diagnostic kits and vaccines against subgroup F adenoviruses. A novel method using sodium dodecyl sulphate SDS/EDTA-pretreated chromatography paper strips was evaluated for the collection, storage and shipping of Ad40/41 contaminated stool samples. Results This study shows that adenoviral DNA can be successfully detected in the filter strips by PCR after four months storage at -20°C, 4°C, room temperature (20–25°C and 37°C. Furthermore no adenoviral infectivity was observed upon contact with the SDS/EDTA-pretreated strips. Conclusions Collecting, storing and transporting adenovirus type 40 and 41 positive stool samples on SDS/EDTA-pretreated chromatography filter strips is a convenient, biosafe and cost effective method for studying new genome variants and monitoring spread of enteric adenovirus strains during outbreaks.

  17. A Cross-Sectional Study on the Perceptions and Practices of Teenagers With Inflammatory Bowel Disease About Repeated Stool Sampling.

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    Heida, Anke; Dijkstra, Alie; Dantuma, Sietske K; van Rheenen, Patrick F

    2016-10-01

    Repeated stool sampling to monitor disease activity is increasingly used in teenagers with inflammatory bowel disease (IBD). Knowledge about their perceptions and practices regarding collection of feces will increase the success rate of this monitoring strategy. We sent a survey to teenagers with IBD treated in an academic center. Seventy-two of 122 invited teenagers completed the survey (response rate 59%; median age 15 years (interquartile range, 13-17). Eighty-five percent reported that stool sampling is normally initiated with help of their parents or caretakers. Seventy-eight percent of respondents say that their parents assist with the placement of stool in the container. Teenagers do not feel embarrassed by the idea of stool sampling, but an active role of the parents or caretakers is an important prerequisite for maintaining a stool-based disease monitoring system. Autonomy in stool sampling is an essential skill required for a successful transition to adult-centered IBD care. Copyright © 2016 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.

  18. Genotype identification of Enterocytozoon bieneusi isolates from stool samples of HIV-infected Tunisian patients

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    Chabchoub N.

    2012-05-01

    Full Text Available The microsporidian species Enterocytozoon bieneusi is a major cause of chronic diarrhea and malabsorption in patients with AIDS. Genotyping was performed on seven E. bieneusi strains for the first time in Tunisia. All the strains were isolated from stool samples of humans with immunodeficiency virus (HIV infection. Analysis of the ribosomal RNA gene internal transcribed spacer (rDNA ITS allowed the identification of three distinct genotypes previously described in other studies. Genotypes D and B were characterized in four and two respectively. The Peruvian genotype (Peru 8 was detected in the last isolate. These results indicate a genetic diversity in E. bieneusi strains from HIV Tunisian patients and suggest the coexistence of both zoonotic and anthroponotic route of transmission.

  19. Parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil: an approach in public health

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    Beatriz Coronato

    2012-04-01

    Full Text Available This research aimed to describe the frequency of parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil. One hundred and five stool samples were collected and processed by the coproparasitological techniques ethyl acetate sedimentation and centrifuge-flotation using saturated sugar solution. Parasites were detected in 81.9% of the samples, hookworm being the most prevalent, followed by Trichuris vulpis. Ascaris sp. eggs were also found. A high level of evolutive forms of parasites with public health risk was found in stool samples of the environment studied. We propose that health education programs, allied to an improvement of human and animal health care, must be employed to reduce the environmental contamination.

  20. Evaluation of Streck tissue fixative, a nonformalin fixative for preservation of stool samples and subsequent parasitologic examination.

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    Nace, E K; Steurer, F J; Eberhard, M L

    1999-12-01

    We undertook a study to evaluate Streck tissue fixative (STF) as a substitute for formalin and polyvinyl alcohol (PVA) in fecal preservation. A comparison of formalin, PVA, (mercuric chloride based), and STF was done by aliquoting fecal samples into each fixative. Stool specimens were collected in Haiti, and parasites included Cyclospora cayetanensis, Giardia intestinalis, Entamoeba coli, Iodamoeba butschlii, Endolimax nana, Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, and Necator americanus. Preserved stools were examined at various predetermined times (1 week, 1 month, and 3 months) to establish the quality of the initial preservation as well as the suitability of the fixative for long-term storage. At each time point, stool samples in fixatives were examined microscopically as follows: (i) in wet mounts (with bright-field and epifluorescence microscopy), (ii) in modified acid-fast-, trichrome-, and safranin-stained smears, and (iii) with two commercial test kits. At the time points examined, morphologic features remained comparable for samples fixed with 10% formalin and STF. For comparisons of STF- and 10% formalin-fixed samples, specific findings showed that Cyclospora oocysts retained full fluorescence, modified acid-fast- and safranin-stained smears of Cryptosporidium and Cyclospora oocysts were equal in staining quality, and results were comparable in the immunofluorescence assay and enzyme immunoassay commercial kits. Stool fixed in STF and stained with trichrome showed less-than-acceptable staining quality compared with stool fixed in PVA. STF provides an excellent substitute for formalin as a fixative in routine examination of stool samples for parasites. However, modifications to the trichrome staining procedures will be necessary to improve the staining quality for protozoal cysts fixed in STF to a level comparable to that with PVA.

  1. Opisthorchis viverrini: Detection by polymerase chain reaction (PCR) in human stool samples

    Digital Repository Service at National Institute of Oceanography (India)

    Umesha, K.R.; SanathKumar; Parvathi, A.; Duenngai, K.; Sithithaworn, P.; Karunasagar, Indrani; Karunasagar, Iddya

    A polymerase chain reaction (PCR) assay was evaluated for detection of Opisthorchis viverrini eggs in the stool specimens of light and heavily infected individuals in Khon Kaen province of Thailand. A total of 75 fecal specimens were analyzed by PCR...

  2. Rapid microsphere assay for identification of cryptosporidium hominis and cryptosporidium parvum in stool and environmental samples.

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    Bandyopadhyay, Kakali; Kellar, Kathryn L; Moura, Iaci; Casaqui Carollo, Maria Cristina; Graczyk, Thaddeus K; Slemenda, Susan; Johnston, Stephanie P; da Silva, Alexandre J

    2007-09-01

    Cryptosporidium hominis and Cryptosporidium parvum are associated with massive disease outbreaks worldwide. Because these two species have different transmission cycles, identification of these parasites to the species level in clinical samples may provide laboratory data of crucial importance in epidemiologic investigations. To date, the most reliable way to differentiate C. hominis and C. parvum is based on DNA sequencing analysis of PCR amplicons. Although this approach is very effective for differentiation of Cryptosporidium species, it is labor-intensive and time-consuming compared with methods that do not require DNA sequencing analysis as an additional step and that have been successfully used for specific identification of a number of pathogens. In this study, we describe a novel Luminex-based assay that can differentiate C. hominis from C. parvum in a rapid and cost-effective manner. The assay was validated by testing a total of 143 DNA samples extracted from clinical specimens, environmental samples, or samples artificially spiked with Cryptosporidium oocysts. As few as 10 oocysts per 300 microl of stools could be detected with this assay. The assay format includes species-specific probes linked to carboxylated Luminex microspheres that hybridize to a Cryptosporidium microsatellite-2 region (ML-2) where C. hominis and C. parvum differ by one nucleotide substitution. The assay proved to be 100% specific when samples that had been characterized by direct fluorescent antibody test (DFA) and DNA sequencing analysis were tested. In addition, the assay was more sensitive than DFA and provided species identification, which is an advantage for epidemiologic studies.

  3. Microbiome analysis of stool samples from African Americans with colon polyps.

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    Brim, Hassan; Yooseph, Shibu; Zoetendal, Erwin G; Lee, Edward; Torralbo, Manolito; Laiyemo, Adeyinka O; Shokrani, Babak; Nelson, Karen; Ashktorab, Hassan

    2013-01-01

    Colonic polyps are common tumors occurring in ~50% of Western populations with ~10% risk of malignant progression. Dietary agents have been considered the primary environmental exposure to promote colorectal cancer (CRC) development. However, the colonic mucosa is permanently in contact with the microbiota and its metabolic products including toxins that also have the potential to trigger oncogenic transformation. To analyze fecal DNA for microbiota composition and functional potential in African Americans with pre-neoplastic lesions. We analyzed the bacterial composition of stool samples from 6 healthy individuals and 6 patients with colon polyps using 16S ribosomal RNA-based phylogenetic microarray; the Human intestinal Tract Chip (HITChip) and 16S rRNA gene barcoded 454 pyrosequencing. The functional potential was determined by sequence-based metagenomics using 454 pyrosequencing. Fecal microbiota profiling of samples from the healthy and polyp patients using both a phylogenetic microarraying (HITChip) and barcoded 454 pyrosequencing generated similar results. A distinction between both sets of samples was only obtained when the analysis was performed at the sub-genus level. Most of the species leading to the dissociation were from the Bacteroides group. The metagenomic analysis did not reveal major differences in bacterial gene prevalence/abundances between the two groups even when the analysis and comparisons were restricted to available Bacteroides genomes. This study reveals that at the pre-neoplastic stages, there is a trend showing microbiota changes between healthy and colon polyp patients at the sub-genus level. These differences were not reflected at the genome/functions levels. Bacteria and associated functions within the Bacteroides group need to be further analyzed and dissected to pinpoint potential actors in the early colon oncogenic transformation in a large sample size.

  4. Microbiome analysis of stool samples from African Americans with colon polyps.

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    Hassan Brim

    Full Text Available Colonic polyps are common tumors occurring in ~50% of Western populations with ~10% risk of malignant progression. Dietary agents have been considered the primary environmental exposure to promote colorectal cancer (CRC development. However, the colonic mucosa is permanently in contact with the microbiota and its metabolic products including toxins that also have the potential to trigger oncogenic transformation.To analyze fecal DNA for microbiota composition and functional potential in African Americans with pre-neoplastic lesions.We analyzed the bacterial composition of stool samples from 6 healthy individuals and 6 patients with colon polyps using 16S ribosomal RNA-based phylogenetic microarray; the Human intestinal Tract Chip (HITChip and 16S rRNA gene barcoded 454 pyrosequencing. The functional potential was determined by sequence-based metagenomics using 454 pyrosequencing.Fecal microbiota profiling of samples from the healthy and polyp patients using both a phylogenetic microarraying (HITChip and barcoded 454 pyrosequencing generated similar results. A distinction between both sets of samples was only obtained when the analysis was performed at the sub-genus level. Most of the species leading to the dissociation were from the Bacteroides group. The metagenomic analysis did not reveal major differences in bacterial gene prevalence/abundances between the two groups even when the analysis and comparisons were restricted to available Bacteroides genomes.This study reveals that at the pre-neoplastic stages, there is a trend showing microbiota changes between healthy and colon polyp patients at the sub-genus level. These differences were not reflected at the genome/functions levels. Bacteria and associated functions within the Bacteroides group need to be further analyzed and dissected to pinpoint potential actors in the early colon oncogenic transformation in a large sample size.

  5. Development and initial evaluation of a lateral flow dipstick test for antigen detection of Entamoeba histolytica in stool sample.

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    Saidin, Syazwan; Yunus, Muhammad Hafiznur; Othman, Nurulhasanah; Lim, Yvonne Ai-Lian; Mohamed, Zeehaida; Zakaria, Nik Zairi; Noordin, Rahmah

    2017-05-01

    Entamoeba histolytica infection remains a public health concern in developing countries. Early diagnosis of amoebiasis can avoid disease complications, thus this study was aimed at developing a test that can rapidly detect the parasite antigens in stool samples. Rabbits were individually immunized with recombinant pyruvate phosphate dikinase (rPPDK) and E. histolytica excretory-secretory antigens to produce polyclonal antibodies. A rapid dipstick test was produced using anti-rPPDK PAb lined on the dipstick as capture reagent and anti-EhESA PAb conjugated to colloidal gold as the detector reagent. Using E. histolytica-spiked in stool sample of a healthy individual, the detection limit of the dipstick test was found to be 1000 cells ml -1 . Meanwhile when rPPDK was spiked in the stool sample, the minimum concentration detected by the dipstick test was 0.1 μg ml -1 . The performances of the dipstick, commercial Techlab E. histolytica II enzyme-linked immunosorbent assays (ELISA) and real-time PCR were compared using 70 stool samples from patients infected with Entamoeba species (n = 45) and other intestinal pathogens (n = 25). When compared to real-time PCR, the diagnostic sensitivity of the dipstick for detection of E. histolytica was 65.4% (n = 17/26); while the diagnostic specificity when tested with stool samples containing other intestinal pathogens was 92% (23/25). In contrast, Techlab E. histolytica II ELISA detected 19.2% (5/26) of the E. histolytica-positive samples as compared to real-time PCR. The lateral flow dipstick test produced in this study enabled rapid detection of E. histolytica, thus it showed good potential to be further developed into a diagnostic tool for intestinal amoebiasis.

  6. Systematic detection and association of Entamoeba species in stool samples from selected sites in India.

    Science.gov (United States)

    Nath, J; Banyal, N; Gautam, D S; Ghosh, S K; Singha, B; Paul, J

    2015-01-01

    This study developed a fast and high throughput dot-blot technique to evaluate the presence of Entamoeba in stool samples (n = 643) followed by a PCR-based method to validate and differentiate the two species E. histolytica and E. dispar. The prevalence rate of the parasite has been detected in a cross-sectional study carried out in the population of the Eastern and Northern parts of India. Of the various demographic features, prevalence was highest in the monsoon season (P = 0·017), in the <15 years age group (P = 0·015). In HIV-positive individuals, the prevalence rate was significantly high (P = 0·008) in patients with a CD4 cell count <200 as well as in patients without antiretroviral therapy (ART) (P = 0·011). Our analysis further confirmed that risk factors such as toilet facilities, living conditions, hygienic practices, drinking water source, occupation and level of education are important predictors as they were found to contribute significantly in the prevalence of the parasite.

  7. Stool C difficile toxin

    Science.gov (United States)

    ... toxin; Colitis - toxin; Pseudomembranous - toxin; Necrotizing colitis - toxin; C difficile - toxin ... be analyzed. There are several ways to detect C difficile toxin in the stool sample. Enzyme immunoassay ( ...

  8. [On the use of FTA technology for collection, archieving, and molecular analysis of microsporidia dna from clinical stool samples].

    Science.gov (United States)

    Sokolova, O I; Dem'ianov, A V; Bovers, L S; Did'e, E S; Sokolova, Iu Ia

    2011-01-01

    The FTA technology was applied for sampling, archiving, and molecular analysis of the DNA isolated from stool samples to diagnose and identify microsporidia, the intracellular opportunistic parasites which induce malabsortion syndrome in immunosuppressed humans, particularly in patients with AIDS. Microsporidia DNA was successfully amplified in 6 of 50 stool samples of HIV-positive patients of the S. P. Botkin Memorial Infectious Disease Hospital (St. Petersburg) applied to FTA cards (FTA-Cars, Whatman Inc. Florham Park, NJ, USA). Amplicons (the fragments of rDNA) were directly sequenced, and microsporidia species--Encephalitozoon intestinalis, E. cuniculi, E. hellem, and Enterocytozoon bieneusi--were identified in Genbank by NCBI BLAST program. The FTA method of DNA immobilization is especially promising for epidemiological and field population studies which involve genotyping of microsporidia species and isolates.

  9. The isolation of Moellerella wisconsensis from stool samples in the U.K.

    Science.gov (United States)

    Marshall, A R; Al-Jumaili, I J; Bint, A J

    1986-01-01

    Three strains of Moellerella wisconsensis were isolated from a total of 400 stool specimens screened for this organism by means of a new selective medium developed in this laboratory. This is the first report of the isolation of this organism in the U.K. The exact role of M. wisconsensis in causing diarrhoea remains to be elucidated.

  10. Antimicrobial susceptibility and β-lactamase production in Bacillus cereus isolates from stool of patients, food and environment samples

    Directory of Open Access Journals (Sweden)

    Savić Dejana

    2016-01-01

    Full Text Available Background/Aim. Bacillus cereus (B. cereus usually ingested by food can cause two types of diseases: vomiting due to the presence of emetic toxin and diarrheal syndrome, due to the presence of diarrheal toxins. Systemic manifestations can also occur. The severe forms of disease demand antibiotic treatmant. The aim of this study was to determine the differences in antibiotic susceptibility and β-lactamase activity of B. cereus isolates from stools of humans, food and environment. Methods. Identification of B. cereus was performed with selective medium, classical biochemical test and polymerase chain reaction (PCR with primers specific for bal gene. Thirty isolates from each group were analysed for antibiotic susceptibility using the disk-diffusion assay. Production of β-lactamase was determined by cefinase test, and double-disc method. Results. All strains identified as B. cereus using classical biochemical test, yielded 533 bp fragment with PCR. Isolates from all the three groups were susceptible to imipenem, vancomycin, and erythromycin. All isolates were susceptible to ciprofloxacin but one from the environment. A statistically significant difference between the groups was confirmed to tetracycline and trimethoprim-sulphamethoxazole sensitivity. A total of 28/30 (93.33% samples from the foods and 25/30 (83.33% samples from environment were approved sensitive to tetracycline, while 10/30 (33.33% isolates from stools were sensitive. Opposite to this result, high susceptibility to trimethoprim-sulphamethoxazole was shown in samples from stools (100%, while isolates from foods (63.33% and from environment (70% had low susceptibility. All samples produced β-lactamases. Conclusion. The strains of B. cereus from all the three groups showed high rate of sensitivity to most tested antibiotics, except to tetracycline in samples from human stool and to trimethoprim-sulphamethoxazole in samples from food and environment. The production of

  11. Prevalence and molecular characterization of human rhinovirus in stool samples of individuals with and without acute gastroenteritis.

    Science.gov (United States)

    Khoonta, Prapaporn; Linsuwanon, Piyada; Posuwan, Nawarat; Vongpunsawad, Sompong; Payungporn, Sunchai; Poovorawan, Yong

    2017-05-01

    Human rhinovirus (RV) most often causes mild upper respiratory tract infection. Although RV is routinely isolated from the respiratory tract, few studies have examined RV in other types of clinical samples. The prevalence of RV was examined in 1,294 stool samples collected mostly from children with acute gastroenteritis residing in Bangkok and Khon Kaen province of Thailand between January 2010 and October 2014. In addition, 591 samples from hand-foot-mouth disease (HFMD) or herpangina patients who do not have gastroenteritis served as a comparison group. Samples were initially screened by semi-nested PCR for the RV 5'UTR through the VP2 capsid region. RV genotyping and phylogenetic analysis were performed on the VP4/VP2 regions. Among children with acute gastroenteritis, RV was found in 2.3% (30/1,294) of stool samples, which comprised 47% (14/30) RV-A, 17% (5/30) RV-B, and 37% (11/30) RV-C. In the comparison group, 0.8% (5/591) was RV-positive and RV-C (3/5) was the major species found. Interestingly, RV was recovered more often from children with acute gastroenteritis than from those with HFMD or herpangina. As many as 31 RV types were present in the gastroenteritis stools, which were different than the types found in those with HFMD or herpangina. J. Med. Virol. 89:801-808, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. Review: Diagnostic accuracy of PCR-based detection tests for Helicobacter Pylori in stool samples.

    Science.gov (United States)

    Khadangi, Fatemeh; Yassi, Maryam; Kerachian, Mohammad Amin

    2017-12-01

    Although different methods have been established to detect Helicobacter pylori (H. pylori) infection, identifying infected patients is an ongoing challenge. The aim of this meta-analysis was to provide pooled diagnostic accuracy measures for stool PCR test in the diagnosis of H. pylori infection. In this study, a systematic review and meta-analysis were carried out on various sources, including MEDLINE, Web of Sciences, and the Cochrane Library from April 1, 1999, to May 1, 2016. This meta-analysis adheres to the guidelines provided by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses report (PRISMA Statement). The clinical value of DNA stool PCR test was based on the pooled false positive, false negative, true positive, and true negative of different genes. Twenty-six of 328 studies identified met the eligibility criteria. Stool PCR test had a performance of 71% (95% CI: 68-73) sensitivity, 96% (95% CI: 94-97) specificity, and 65.6 (95% CI: 30.2-142.5) diagnostic odds ratio (DOR) in diagnosis of H. pylori. The DOR of genes which showed the highest performance of stool PCR tests was as follows: 23S rRNA 152.5 (95% CI: 55.5-418.9), 16S rRNA 67.9 (95%CI: 6.4-714.3), and glmM 68.1 (95%CI: 20.1-231.7). The sensitivity and specificity of stool PCR test are relatively in the same spectrum of other diagnostic methods for the detection of H. pylori infection. In descending order of significance, the most diagnostic candidate genes using PCR detection were 23S rRNA, 16S rRNA, and glmM. PCR for 23S rRNA gene which has the highest performance could be applicable to detect H. pylori infection. © 2017 John Wiley & Sons Ltd.

  13. Real-time PCR using SYBR Green for the detection of Shigella spp. in food and stool samples.

    Science.gov (United States)

    Mokhtari, W; Nsaibia, S; Gharbi, A; Aouni, M

    2013-02-01

    Shigella spp are exquisitely fastidious Gram negative organisms that frequently get missed in the detection by traditional culture methods. For this reason, this work has adapted a classical PCR for detection of Shigella in food and stool specimens to real-time PCR using the SYBR Green format. This method follows a melting curve analysis to be more rapid and provide both qualitative and quantitative data about the targeted pathogen. A total of 117 stool samples with diarrhea and 102 food samples were analyzed in Public Health Regional Laboratory of Nabeul by traditional culture methods and real-time PCR. To validate the real-time PCR assay, an experiment was conducted with both spiked and naturally contaminated stool samples. All Shigella strains tested were ipaH positive and all non-Shigella strains yielded no amplification products. The melting temperature (T(m) = 81.5 ± 0.5 °C) was consistently specific for the amplicon. Correlation coefficients of standard curves constructed using the quantification cycle (C(q)) versus copy numbers of Shigella showed good linearity (R² = 0.995; slope = 2.952) and the minimum level of detection was 1.5 × 10³ CFU/g feces. All food samples analyzed were negative for Shigella by standard culture methods, whereas ipaH was detected in 8.8% culture negative food products. Moreover, the ipaH specific PCR system increased the detection rate over that by culture alone from 1.7% to 11.1% among patients with diarrhea. The data presented here shows that the SYBR Green I was suitable for use in the real-time PCR assay, which provided a specific, sensitive and efficient method for the detection and quantification of Shigella spp in food and stool samples. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Electron microscopy identification of microsporidia (enterocytozoon bieneusi and cyclospora cayetanensis from stool samples of HIV infected patients

    Directory of Open Access Journals (Sweden)

    Satheeshkumar S

    2004-01-01

    Full Text Available Microsporidia (Enterocytozoon bieneusi and Cyclospora cayetanensis have been reported worldwide causing diarrhoea in AIDS patients. Stool samples from HIV infected patients were subjected to routine examination for parasites, followed by special staining techniques to detect microsporidia and Cyclospora cayetanensis. Confirmed positive cases of these parasites were further processed for electron microscopy identity of the parasites and characteristic details. Scanning and transmission electron microscopy showed better morphological and structural details of the parasites.

  15. Impact of changing from staining to culture techniques on detection rates of Campylobacter spp. in routine stool samples in Chile.

    Science.gov (United States)

    Porte, Lorena; Varela, Carmen; Haecker, Thomas; Morales, Sara; Weitzel, Thomas

    2016-05-13

    Campylobacter is a leading cause of bacterial gastroenteritis, but sensitive diagnostic methods such as culture are expensive and often not available in resource limited settings. Therefore, direct staining techniques have been developed as a practical and economical alternative. We analyzed the impact of replacing Campylobacter staining with culture for routine stool examinations in a private hospital in Chile. From January to April 2014, a total of 750 consecutive stool samples were examined in parallel by Hucker stain and Campylobacter culture. Isolation rates of Campylobacter were determined and the performance of staining was evaluated against culture as the gold standard. Besides, isolation rates of Campylobacter and other enteric pathogens were compared to those of past years. Campylobacter was isolated by culture in 46 of 750 (6.1 %) stool samples. Direct staining only identified three samples as Campylobacter positive and reached sensitivity and specificity values of 6.5 and 100 %, respectively. In comparison to staining-based detection rates of previous years, we observed a significant increase of Campylobacter cases in our patients. Direct staining technique for Campylobacter had a very low sensitivity compared to culture. Staining methods might lead to a high rate of false negative results and an underestimation of the importance of campylobacteriosis. With the inclusion of Campylobacter culture, this pathogen became a leading cause of intestinal infection in our patient population.

  16. Evaluation of Helicobacter pylori antigen positivity in stool samples of patients with dyspeptic complaints in a tertiary care hospital

    Directory of Open Access Journals (Sweden)

    Mehmet Burak Selek

    2013-12-01

    Full Text Available Objective: Helicobacter pylori is a microorganism associatedwith gastritis, peptic ulcer disease and gastriccancer. We aimed to figure out the positivity rate in stoolsamples of outpatients with dyspeptic complaints visitinggastroenterology department and to evaluate its relationwith age, gender and seasonal changes.Methods: Between January 01, 2012 and December 31,2012, stool samples of 330 adult outpatients admitted togastroenterology department are investigated with an immunochromatographictest kit using monoclonal antibodiesfor detection of H. pylori antigen.Results: Among 330 patients’ stool samples tested, 67(20.3% were positive. 18.6% of men and 22.2% of womenwere detected as positive. According to age groups,17.1% patients were positive for 15-35 age groups,27.1% patients were positive for 36-55 age groups and18.2% patients were positive for above 56. Seasonal differenceof H. pylori antigen positivity in stool samples wasstatistically significant (p=0.001. Highest positivity rate29.7% was detected for winter months (December-January-February. According to logistic regression analysis,winter is found as a risk factor with statistically significant2.295 times greater risk [p=0001, Exp (B = 2.925, 95.0%C.I. for EXP (B = 1.668-5.129].Conclusion: H. pylori antigen positivity rate of our study islower than other previously conducted studies in Turkey.But, positivity rates are higher among women comparedto men, concordant with other studies. Even more, detectionof high positivity rates in winter shows primary infectionand/or relapse can be affected by seasonal changes.Key words: Helicobacter pylori, gastroenterology, stool antigen test

  17. Generation of whole genome sequences of new Cryptosporidium hominis and Cryptosporidium parvum isolates directly from stool samples.

    Science.gov (United States)

    Hadfield, Stephen J; Pachebat, Justin A; Swain, Martin T; Robinson, Guy; Cameron, Simon Js; Alexander, Jenna; Hegarty, Matthew J; Elwin, Kristin; Chalmers, Rachel M

    2015-08-29

    Whole genome sequencing (WGS) of Cryptosporidium spp. has previously relied on propagation of the parasite in animals to generate enough oocysts from which to extract DNA of sufficient quantity and purity for analysis. We have developed and validated a method for preparation of genomic Cryptosporidium DNA suitable for WGS directly from human stool samples and used it to generate 10 high-quality whole Cryptosporidium genome assemblies. Our method uses a combination of salt flotation, immunomagnetic separation (IMS), and surface sterilisation of oocysts prior to DNA extraction, with subsequent use of the transposome-based Nextera XT kit to generate libraries for sequencing on Illumina platforms. IMS was found to be superior to caesium chloride density centrifugation for purification of oocysts from small volume stool samples and for reducing levels of contaminant DNA. The IMS-based method was used initially to sequence whole genomes of Cryptosporidium hominis gp60 subtype IbA10G2 and Cryptosporidium parvum gp60 subtype IIaA19G1R2 from small amounts of stool left over from diagnostic testing of clinical cases of cryptosporidiosis. The C. parvum isolate was sequenced to a mean depth of 51.8X with reads covering 100 % of the bases of the C. parvum Iowa II reference genome (Bioproject PRJNA 15586), while the C. hominis isolate was sequenced to a mean depth of 34.7X with reads covering 98 % of the bases of the C. hominis TU502 v1 reference genome (Bioproject PRJNA 15585). The method was then applied to a further 17 stools, successfully generating another eight new whole genome sequences, of which two were C. hominis (gp60 subtypes IbA10G2 and IaA14R3) and six C. parvum (gp60 subtypes IIaA15G2R1 from three samples, and one each of IIaA17G1R1, IIaA18G2R1, and IIdA22G1), demonstrating the utility of this method to sequence Cryptosporidium genomes directly from clinical samples. This development is especially important as it reduces the requirement to propagate

  18. Isolation of pathogenic Escherichia coli from stool samples of diarrhoeal patients with history of raw milk consumption

    Directory of Open Access Journals (Sweden)

    M. N. Brahmbhatt

    2013-07-01

    Full Text Available Aim: To detect the occurrence of pathogenic Escherichia coli from stool samples of diarrhoeal patients with history of raw milk consumption and to determine the public health significance of isolates, especially their role in causing human diseases.Materials and Methods: Atotal of 100 stool samples from diarrhoeal patients, with history of raw milk consumption were collected from primary health centres in and around Anand city, under aseptic conditions and a total of 50 raw milk samples were collected from milk vendors, retail shops located in Anand city in sterilized sample bottles. MacConkey broth was used for the enrichment of all the samples and inoculation was done on MacConkey agar and EMB agar was used as the selective media. This was followed by the confirmation of isolates using biochemical tests. For the serotyping,E. coli isolates were sent to the National Salmonella and Escherichia Centre, Central Research Institute (CRI, Kasauli, Himachal Pradesh.Detection of virulence genes was performed using PCR technique.Results: During the present investigation, 26 (52% E. coli isolates from 50 milk samples and 59 (59% E. coli isolates from 100 stool samples were recovered. Out of 85 E. coli isolates sent for serotyping, 74 isolates could be typed which were further distributed into 13 different serogroups O2, O4, O8, O17, O22, O25, O29, O36, O45, O60, O90, O116 and O172, whereas 8 isolates were found untypable and 3 isolates were reported rough isolates. Of the 59 E. coli isolates from stool samples of diarrhoeal patients tested, 15 isolates (25.42% were reported to be positive for stx genes, among that 6 (10.16% were positive for stx1 gene, 9 (15.25% isolates were positive for stx2 gene, while 3 isolates (5.08% were positive for eaeA gene. In this study, 21 E. coliisolates were found to be Shiga toxin producing E. coli (STEC while none of the isolates were positive for the serotype O157. Conclusions: Our present findings indicate that raw

  19. Improved amplification efficiency on stool samples by addition of spermidine and its use for non-invasive detection of colorectal cancer

    KAUST Repository

    Roperch, Jean-Pierre

    2015-05-29

    Background Using quantitative methylation-specific PCR (QM-MSP) is a promising method for colorectal cancer (CRC) diagnosis from stool samples. Difficulty in eliminating PCR inhibitors of this body fluid has been extensively reported. Here, spermidine is presented as PCR facilitator for the detection of stool DNA methylation biomarkers using QM-MSP. We examined its effectiveness with NPY, PENK and WIF1, three biomarkers which we have previously shown to be of relevance to CRC. Results We determined an optimal window for the amplification of the albumin (Alb) gene (100 ng of bisulfite-treated stool DNA added of 1 mM spermidine) at which we report that spermidine acts as a PCR facilitator (AE = 1680%) for SG RT-PCR. We show that the amplification of methylated PENK, NPY and WIF1 is considerably facilitated by QM-MSP as measured by an increase of CMI (Cumulative Methylation Index, i.e. the sum of the three methylation values) by a factor of 1.5 to 23 fold in individual samples, and of 10 fold in a pool of five samples. Conclusions We contend that spermidine greatly reduces the problems of PCR inhibition in stool samples. This observed feature, after validation on a larger sampling, could be used in the development of stool-based CRC diagnosis tests.

  20. Stool Softeners

    Science.gov (United States)

    ... any part you do not understand. Take stool softeners exactly as directed. Do not take more or less of it or take it more often than prescribed by your doctor.Take capsules and tablets with a full glass of water. The liquid comes with a specially marked dropper ...

  1. Identification of human Norovirus (HNoV in domestic pig stool samples

    Directory of Open Access Journals (Sweden)

    María F. Gutiérrez

    2011-08-01

    Full Text Available To determine the presence of NoVs as a possible causal zoonotic agent of acute diarrhea in pigs and humans. Materialsand methods. We collected a total of 77 samples from diarrheal children under 5 years and pigs under 2 months from La Chambatown in Tolima, Colombia. These samples were transported to the Laboratory of Virology of the Pontificia Universidad Javerianain Bogotá, and extraction with Trizol-reagent was done following the manufacturer’s instructions. After obtaining the RNA, thenext step was to perform RT-PCR for obtaining the expected amplification product of 213- bp NoVs. Finally, the positive samplesobtained in the RT-PCR were sequenced and analyzed by bioinformatics methods. Results. Six positive diarrheic samples fromchildren and a positive diarrheic sample from pigs were detected by a band of 231 bp. Five of the six positive samples in childrenand the positive pig sample were sequenced and analyzed. Conclusion. Given the close genetic relationship between pig andhuman sequences, this could be an indication of the potential existence of a common animal acting as a reservoir for human orother animal strains.

  2. Efficiency of Direct Microscopy of Stool Samples Using an Antigen-Specific Adhesin Test for Entamoeba Histolytica

    Directory of Open Access Journals (Sweden)

    Arzu İrvem

    2016-10-01

    Full Text Available Background: E. histolytica is among the common causes of acute gastroenteritis. The pathogenic species E. histolytica and the nonpathogenic species E. dispar cannot be morphologically differentiated, although correct identification of these protozoans is important for treatment and public health. In many laboratories, the screening of leukocytes, erythrocytes, amoebic cysts, trophozoites and parasite eggs is performed using Native-Lugol’s iodine for pre-diagnosis. Aims: In this study, we aimed to investigate the frequency of E. histolytica in stool samples collected from 788 patients residing in the Anatolian region of İstanbul who presented with gastrointestinal complaints. We used the information obtained to evaluate the effectiveness of microscopic examinations when used in combination with the E. histolytica adhesin antigen test. Study Design: Retrospective cross-sectional study Methods: Preparations of stool samples stained with Native-Lugol’s iodine were evaluated using the E. histolytica adhesin test and examined using standard light microscopy at ×40 magnification. Pearson’s Chi-square and Fisher’s exact tests were used for statistical analysis. Logistic regression analysis was used for multivariate analysis. Results: Of 788 samples, 38 (4.8% were positive for E. histolytica adhesin antigens. When evaluated together with the presences of erythrocytes, leukocytes, cysts, and trophozoites, respectively, using logistic regression analysis, leukocyte positivity was significantly higher. The odds ratio of leukocyte positivity increased adhesin test-positivity by 2,530-fold (95% CI=1.01–6.330. Adhesin test-positivity was significant (p=0.047. Conclusion: In line with these findings, the consistency between the presence of cysts and erythrocytes and adhesin test-positivity was found to be highly significant, but that of higher levels of leukocytes was found to be discordant. It was concluded that leukocytes and trophozoites were

  3. Efficiency of Direct Microscopy of Stool Samples Using an Antigen-Specific Adhesin Test for Entamoeba Histolytica.

    Science.gov (United States)

    İrvem, Arzu; Özdil, Kamil; Çalışkan, Zuhal; Yücel, Muhterem

    2016-09-01

    E. histolytica is among the common causes of acute gastroenteritis. The pathogenic species E. histolytica and the nonpathogenic species E. dispar cannot be morphologically differentiated, although correct identification of these protozoans is important for treatment and public health. In many laboratories, the screening of leukocytes, erythrocytes, amoebic cysts, trophozoites and parasite eggs is performed using Native-Lugol's iodine for pre-diagnosis. In this study, we aimed to investigate the frequency of E. histolytica in stool samples collected from 788 patients residing in the Anatolian region of İstanbul who presented with gastrointestinal complaints. We used the information obtained to evaluate the effectiveness of microscopic examinations when used in combination with the E. histolytica adhesin antigen test. Retrospective cross-sectional study. Preparations of stool samples stained with Native-Lugol's iodine were evaluated using the E. histolytica adhesin test and examined using standard light microscopy at ×40 magnification. Pearson's Chi-square and Fisher's exact tests were used for statistical analysis. Logistic regression analysis was used for multivariate analysis. Of 788 samples, 38 (4.8%) were positive for E. histolytica adhesin antigens. When evaluated together with the presences of erythrocytes, leukocytes, cysts, and trophozoites, respectively, using logistic regression analysis, leukocyte positivity was significantly higher. The odds ratio of leukocyte positivity increased adhesin test-positivity by 2,530-fold (95% CI=1.01-6.330). Adhesin test-positivity was significant (p=0.047). In line with these findings, the consistency between the presence of cysts and erythrocytes and adhesin test-positivity was found to be highly significant, but that of higher levels of leukocytes was found to be discordant. It was concluded that leukocytes and trophozoites were easily misjudged using direct microscopy. Although microscopic examination of samples

  4. Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea

    DEFF Research Database (Denmark)

    Mero, Sointu; Kirveskari, Juha; Antikainen, Jenni

    2017-01-01

    Background: In developing countries, diarrhoea is the most common cause of death for children under five years of age, with Giardia lamblia, Cryptosporidium and Entamoeba histolytica as the most frequent pathogenic parasites. Traditional microscopy for stool parasites has poor sensitivity...... was developed for detecting Giardia lamblia, Cryptosporidium hominis/parvum and Entamoeba histolytica directly from stool specimens. Applicability to dried samples was checked by comparing with fresh ones in a small test material. Finally, the assay was applied to dried specimens collected from Guinea......-Bissauan children with diarrhoea. Results: The PCR’s analytical sensitivity limit was 0.1 ng/ml for G. lamblia DNA, 0.01 ng/ml for E. histolytica DNA and 0.1 ng/ml for Cryptosporidium sp. In the test material, the assay performed similarly with fresh and dried stools. Of the 52 Guinea-Bissauan samples, local...

  5. An uncooked vegan diet shifts the profile of human fecal microflora: computerized analysis of direct stool sample gas-liquid chromatography profiles of bacterial cellular fatty acids.

    Science.gov (United States)

    Peltonen, R; Ling, W H; Hänninen, O; Eerola, E

    1992-01-01

    The effect of an uncooked extreme vegan diet on fecal microflora was studied by direct stool sample gas-liquid chromatography (GLC) of bacterial cellular fatty acids and by quantitative bacterial culture by using classical microbiological techniques of isolation, identification, and enumeration of different bacterial species. Eighteen volunteers were divided randomly into two groups. The test group received an uncooked vegan diet for 1 month and a conventional diet of mixed Western type for the other month of the study. The control group consumed a conventional diet throughout the study period. Stool samples were collected. Bacterial cellular fatty acids were extracted directly from the stool samples and measured by GLC. Computerized analysis of the resulting fatty acid profiles was performed. Such a profile represents all bacterial cellular fatty acids in a sample and thus reflects its microflora and can be used to detect changes, differences, or similarities of bacterial flora between individual samples or sample groups. GLC profiles changed significantly in the test group after the induction and discontinuation of the vegan diet but not in the control group at any time, whereas quantitative bacterial culture did not detect any significant change in fecal bacteriology in either of the groups. The results suggest that an uncooked extreme vegan diet alters the fecal bacterial flora significantly when it is measured by direct stool sample GLC of bacterial fatty acids. PMID:1482187

  6. Cyclospora cayetanensis in sputum and stool samples Cyclospora cayetanensis em amostra de escarro

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    Angela Beatriz DI GLIULLO

    2000-04-01

    Full Text Available We report the observation of acid-fast Cyclospora cayetanensis oocysts in a sputum sample. The patient, a 60 year-old, HIV negative man, was successfully treated for pulmonary tuberculosis during 1997. On February 1998, he was admitted to our center due to loss of weight, cough with purulent expectoration, dysphonia and a radiological picture of pulmonary fibrosis. Bacilloscopic study of sputum (negative for acid-fast bacilli stained with Ziehl-Neelsen technique showed large (8-10 µm spherical, acid-fast Cyclospora cayetanensis oocysts. No other pathogens were isolated on cultures from this sample or from laryngeal biopsy. Serial parasitologic studies showed C. cayetanensis and also eggs of Trichuris trichiura, Ascaris lumbricoides and Hymenolepis nana and of Entamoeba coli cysts. The patient lives in the outskirts of Buenos Aires in a brick-made house with potable water and works as builder of sewers. He travelled in several occasions to the rural area of province of Tucumán which has poor sanitary conditions. C. cayetanensis is an emergent agent of diarrhea and as far as we know this is the first time the parasite is observed in respiratory samples.Comunicamos a observação de grandes oocistos (8-10 µm de diâmetro esféricos, ácido-álcool-resistentes de Cyclospora cayetanensis em amostra de escarro corada com a técnica de Ziehl-Neelsen. Na amostra não foram observados nem cultivados outros agentes patogênicos. Trata-se de um paciente do sexo masculino, 60 anos de idade, HIV (-, tratado previamente para tuberculose pulmonar (1997. Em fevereiro de 1998 apresentou-se em nosso hospital com perda de peso, tosse com expectoração purulenta, disfonia e imagens radiológicas de fibrose pulmonar. As culturas das amostras de escarro e da biopsia de laringe foram negativas. O exame parasitológico seriado de fezes mostrou ovos de Ascaris lumbricoides, Hymenolepis nana e Trichuris trichiura e cistos de Entamoeba coli. O paciente mora nos

  7. Two sequential PCR amplifications for detection of Schistosoma mansoni in stool samples with low parasite load

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    Maria Cristina Carvalho do Espírito-Santo

    2012-10-01

    Full Text Available Schistosomiasis constitutes a major public health problem, with an estimated 200 million individuals infected worldwide and 700 million people living in risk areas. In Brazil there are areas of high, medium and low endemicity. Studies have shown that in endemic areas with a low prevalence of Schistosoma infection the sensitivity of parasitological methods is clearly reduced. Consequently diagnosis is often impeded due to the presence of false-negative results. The aim of this study is to present the PCR reamplification (Re-PCR protocol for the detection of Schistosoma mansoni in samples with low parasite load (with less than 100 eggs per gram (epg of feces. Three methods were used for the lysis of the envelopes of the S. mansoni eggs and two techniques of DNA extraction were carried out. Extracted DNA was quantified, and the results suggested that the extraction technique, which mixed glass beads with a guanidine isothiocyanate/phenol/chloroform (GT solution, produced good results. PCR reamplification was conducted and detection sensitivity was found to be five eggs per 500 mg of artificially marked feces. The results achieved using these methods suggest that they are potentially viable for the detection of Schistosoma infection with low parasite load.

  8. DETECTION OF HELICOBACTER ANTIGEN IN STOOL SAMPLES AND ITS RELATION TO H. PYLORI POSITIVE CHOLECYSTITIS IN EGYPTIAN PATIENTS WITH CHRONIC CALCULAR CHOLECYSTITIS.

    Science.gov (United States)

    Hassan, Ehsan H; Gerges, Shawkat S; Ahmed, Rehab; Mostafa, Zeinab M; Al-Hamid, Hager Abd; Abd El-Galil, Heba; Thabet, Suzan

    2015-12-01

    Evidences supporting the association between H. pylori infection and chronic cholecystitis could be found by using direct culture or staining of H. pylori in gallbladder tissues as well as indirect techniques. Stool antigen test has been widely used due to its noninvasive nature. Various stool antigen tests were developed to detect H. pylori using an enzyme immunoassay (EIA) based on monoclonal or polyclonal antibodies This study evaluated the frequency of H. pylori antigen in stool samples of patients with chronic calcular cholecystitis as regard gall bladder histopathological changes. Fifty patients were included presented with symptomatic qholecystolithiasis recruited from the outpatient clinic of National Hepatology and Tropical Medicine Research Institute during 2014-2015. Full history and clinical examination and abdominal ultrasonography were performed. Stool samples were collected, prepared and examined for detection of H. pylori antigen. Cholecystectomy was done for all patients; 45 patients (90%) by laparoscopic Cholecystectomy and 5 patients (10%) by open surgery and removed gallbladders were submitted to pathology department for detection of H. pylori in tissue under microscope using Giemsa stain. The results showed that (82%) were females with mean age (42.6 +/- 1 years). The mean BMI was (29 + 7.2) H. pylori-specific antigen in stool samples was detected in 40% of patients and 38% were detected in patients; tissue, with significant correlation between H. pylori-specific antigen in stool and in tissue. Histopathological pictures infection in tissue were 68.4% mucosal erosions, 63.2% mucosal atrophy, 57.9% mucosal hyperplasia, 26.3% metaplasia, 42.1% musculosa hypertrophy, 26.3% fibrosis, but lymphoid aggregates were in 42.1% of cases.

  9. Detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in clinical stool samples by using multiplex real-time PCR after automated DNA isolation

    NARCIS (Netherlands)

    Van Lint, P; Rossen, J W; Vermeiren, S; Ver Elst, K; Weekx, S; Van Schaeren, J; Jeurissen, A

    2013-01-01

    Diagnosis of intestinal parasites in stool samples is generally still carried out by microscopy; however, this technique is known to suffer from a low sensitivity and is unable to discriminate between certain protozoa. In order to overcome these limitations, a real-time multiplex PCR was evaluated

  10. Differentiation of Entamoeba histolytica/Entamoeba dispar by the polymerase chain reaction in stool samples of patients with gastrointestinal symptoms in the Sanliurfa Province.

    Science.gov (United States)

    Zeyrek, Fadile Yıldız; Turgay, Nevin; Unver, Aysegül; Ustün, Sebnem; Akarca, Ulus; Töz, Seray

    2013-01-01

    We aimed to diagnose amebiasis and also identify Entamoeba histolytica (E. histolytica) and Entamoeba dispar (E. dispar) in patients with gastrointestinal symptoms in an endemic region in Turkey. Stool samples obtained from 181 patients with gastrointestinal symptoms from the Harran University Hospital of Sanliurfa were examined for the diagnosis of amebiasis by the three methods which are as follows:- In house polymerase chain reaction (PCR) targeting the 135 base pair region located on the small-subunit ribosomal RNA (SSU rRNA) gene to differentiate E. histolytica from E. dispar; and the commercial kit, RIDASCREEN® stool ELISA, that identifies Entamoeba sensu lato antigen and microscopical examination of Trichrome stained smears of stool samples. Positivity for E. histolytica/E. dispar complex was found to be 79 (43.6%) by microscopy versus 83 (45.9%) by PCR out of 181 stool samples. A total of 45 patients were found to be positive by the antigen detection method. PCR and microscopy were both positive in 59 samples. The number of patients infected with E. dispar (39.8%) was found to be higher than E. histolytica (3.3%) while 5 patients (2.8%) had mixed E. histolytica+E. dispar infections according to PCR results. Routine diagnosis of amebiasis by a combination of microscopy and antigen detection technique should be complemented with a PCR assay as a reference test for sensitive differentiation of both species.

  11. Molecular typing and virulence analysis of serotype K1 Klebsiella pneumoniae strains isolated from liver abscess patients and stool samples from noninfectious subjects in Hong Kong, Singapore, and Taiwan.

    Science.gov (United States)

    Siu, L Kristopher; Fung, Chang-Phone; Chang, Feng-Yee; Lee, Nelson; Yeh, Kuo-Ming; Koh, Tse Hsien; Ip, Margaret

    2011-11-01

    Serotype K1 Klebsiella pneumoniae with multilocus sequence type 23 (ST23) has been strongly associated with liver abscess in Taiwan. Few data regarding the strain types and virulence of this serotype from other Asian countries are available. Serotype K1 K. pneumoniae strains isolated from liver abscess and stool samples from subjects hospitalized in Hong Kong, Singapore, and Taiwan hospitals were examined. Forty-seven serotype K1 isolates were identified: 26 from liver abscess samples and 21 from stool samples. MLST revealed 7 sequence types: 85.1% (40 of 47 isolates) belonged to ST23, 1 isolate belonged to ST163 (a single-locus variant of ST23), and 2 isolates were ST249 (a 3-locus variant of ST23). New STs, namely, ST367, ST425, and ST426, were allocated to 3 of 4 isolates from stool samples. The virulence of these strains was determined by neutrophil phagocytosis and mouse infection models. Except for two ST23 isolates, all Klebsiella pneumoniae isolates were resistant to phagocytosis. Resistance to serum killing varied in isolates of ST23, while all non-ST23 strains were susceptible to serum killing except one with ST249 from a liver abscess. All hypervirulent isolates with a 50% lethal dose of serum killing, and also carried both virulence-associated genes, rmpA and aerobactin. Multilocus sequence typing genotype 23 was the most prevalent sequence type among serotype K1 K. pneumoniae isolates from both liver abscess and stool samples in the Asia Pacific region. Serotype K1 K. pneumoniae isolates with capsule expression leading to phagocytic resistance and with the aerobactin gene were associated with hypervirulence.

  12. Antibiotic resistance among Escherichia coli isolates from stool samples of children aged 3 to 14 years from Ujjain, India

    Science.gov (United States)

    2013-01-01

    Background Antibiotic resistance is a major global public health concern, particularly in settings where few treatment options are available. Limited research has been done on antibiotic resistance in Escherichia coli of Indian children at community level. Therefore we studied antibiotic resistance patterns in E. coli isolates from stool samples of children aged 3-14 years from Ujjain, Central India, to investigate associations of resistance with demographic variables. Methods Children, 3-14 years of age, were included from 30 randomly selected villages of Palwa demographic surveillance site, Ujjain, India. Parents were interviewed using a questionnaire, and stool samples were collected from participating children. E. coli were isolated from stool samples (n = 529), and susceptibility testing to 18 different antibiotics was done using standard methods. Results The proportions of isolates resistant to various antibiotics were, nalidixic acid, (45%), tetracycline (37%), ampicillin (37%), sulfamethoxazole/trimethoprim (29%) and amoxicillin/clavulanic acid (29%). No isolates were resistant to imipenem. Overall, 72% of isolates were resistant to at least one antibiotic and 33% were multi-drug resistant. High rates of cross-resistance were seen for 15 (83%) of the antibiotics studied. E. coli isolates from children with literate mothers were more resistant to penicillins and fluoroquinolones. ESBL-producers comprised 9% of the isolates. Conclusion Antibiotic resistance and cross-resistance were common in E. coli from stools of children. Resistance rates were associated with maternal literacy. PMID:24124728

  13. Evaluation of sampling and storage procedures on preserving the community structure of stool microbiota: A simple at-home toilet-paper collection method.

    Science.gov (United States)

    Al, Kait F; Bisanz, Jordan E; Gloor, Gregory B; Reid, Gregor; Burton, Jeremy P

    2018-01-01

    The increasing interest on the impact of the gut microbiota on health and disease has resulted in multiple human microbiome-related studies emerging. However, multiple sampling methods are being used, making cross-comparison of results difficult. To avoid additional clinic visits and increase patient recruitment to these studies, there is the potential to utilize at-home stool sampling. The aim of this pilot study was to compare simple self-sampling collection and storage methods. To simulate storage conditions, stool samples from three volunteers were freshly collected, placed on toilet tissue, and stored at four temperatures (-80, 7, 22 and 37°C), either dry or in the presence of a stabilization agent (RNAlater®) for 3 or 7days. Using 16S rRNA gene sequencing by Illumina, the effect of storage variations for each sample was compared to a reference community from fresh, unstored counterparts. Fastq files may be accessed in the NCBI Sequence Read Archive: Bioproject ID PRJNA418287. Microbial diversity and composition were not significantly altered by any storage method. Samples were always separable based on participant, regardless of storage method suggesting there was no need for sample preservation by a stabilization agent. In summary, if immediate sample processing is not feasible, short term storage of unpreserved stool samples on toilet paper offers a reliable way to assess the microbiota composition by 16S rRNA gene sequencing. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Detection of Campylobacter species and Arcobacter butzleri in stool samples by use of real-time multiplex PCR.

    Science.gov (United States)

    de Boer, Richard F; Ott, Alewijn; Güren, Pinar; van Zanten, Evert; van Belkum, Alex; Kooistra-Smid, Anna M D

    2013-01-01

    The presence of Campylobacter (or Campylobacter-like) species in stools from patients suspected of infectious gastroenteritis (n = 493) was investigated using real-time PCR for detection of Arcobacter butzleri (hsp60 gene), Campylobacter coli (ceuE gene), Campylobacter jejuni (mapA), five acknowledged pathogenic Campylobacter spp. (C16S_Lund assay), and the Campylobacter genus (C16S_LvI assay). In total, 71.4% of the samples were positive for Campylobacter DNA (n = 352) by a Campylobacter genus-specific (C16S_LvI) assay. A total of 23 samples (4.7%) were positive in the C16S_Lund assay, used for detection of C. jejuni, C. coli, C. lari, C. upsaliensis, and C. hyointestinalis. Subsequent identification of these samples yielded detection frequencies (DF) of 4.1% (C. jejuni), 0.4% (C. coli), and 0.4% (C. upsaliensis). The DF of A. butzleri was 0.4%. Interestingly, sequencing of a subgroup (n = 46) of C16S_LvI PCR-positive samples resulted in a considerable number of Campylobacter concisus-positive samples (n = 20). PCR-positive findings with the C16S_Lund and C. jejuni/C. coli-specific assays were associated with more serious clinical symptoms (diarrhea and blood). Threshold cycle (C(T)) values of C. jejuni/C. coli PCR-positive samples were comparable to those of the C16S_Lund PCR (P = 0.21). C(T) values for both assays were significantly lower than those of the C16S_LvI assay (P < 0.001 and P < 0.00001, respectively). In conclusion, this study demonstrated that in combination, the C. jejuni/C coli-specific assays and the C16S_Lund assay are both useful for routine screening purposes. Furthermore, the DF of the emerging pathogen C. concisus was at least similar to the DF of C. jejuni.

  15. A conventional polymerase chain reaction-based method for the diagnosis of human schistosomiasis in stool samples from individuals in a low-endemicity area

    Directory of Open Access Journals (Sweden)

    Teiliane Rodrigues Carneiro

    2013-12-01

    Full Text Available The aim of this study was to evaluate the efficacy of a polymerase chain reaction (PCR-based method to detect Schistosoma mansoni DNA in stool samples from individuals living in a low-endemicity area in Brazil. Of the 125 initial stool samples, 80 were ELISA reactive and eggs were identified in 19 of the samples by parasitological examination. For the PCR evaluations, 56 stool samples were selected and divided into five groups. Groups I-IV were scored negative for S. mansoni eggs by parasitological examination. Groups I and II were ELISA reactive, whereas Groups III and IV were ELISA nonreactive. Groups II and III were positive for other intestinal parasites. PCR testing scored eight samples as positive from these four groups. Group V represented the S. mansoni -positive group and it included ELISA-reactive samples that were scored positive for S. mansoni by one or more parasitological examinations (6/19 were positive by Kato-Katz method, 9/17 by saline gradient and 10/13 by Helmintex®. PCR scored 13 of these 19 samples as positive for S. mansoni . We conclude that while none of these methods yielded 100% sensitivity, a combination of techniques should be effective for improving the detection of S. mansoni infection in low-endemicity areas.

  16. Reaction mechanism of Ru(II) piano-stool complexes: umbrella sampling QM/MM MD study.

    Science.gov (United States)

    Futera, Zdeněk; Burda, Jaroslav V

    2014-07-15

    Biologically relevant interactions of piano-stool ruthenium(II) complexes with ds-DNA are studied in this article by hybrid quantum mechanics-molecular mechanics (QM/MM) computational technique. The whole reaction mechanism is divided into three phases: (i) hydration of the [Ru(II) (η(6) -benzene)(en)Cl](+) complex, (ii) monoadduct formation between the resulting aqua-Ru(II) complex and N7 position of one of the guanines in the ds-DNA oligomer, and (iii) formation of the intrastrand Ru(II) bridge (cross-link) between two adjacent guanines. Free energy profiles of all the reactions are explored by QM/MM MD umbrella sampling approach where the Ru(II) complex and two guanines represent a quantum core, which is described by density functional theory methods. The combined QM/MM scheme is realized by our own software, which was developed to couple several quantum chemical programs (in this study Gaussian 09) and Amber 11 package. Calculated free energy barriers of the both ruthenium hydration and Ru(II)-N7(G) DNA binding process are in good agreement with experimentally measured rate constants. Then, this method was used to study the possibility of cross-link formation. One feasible pathway leading to Ru(II) guanine-guanine cross-link with synchronous releasing of the benzene ligand is predicted. The cross-linking is an exergonic process with the energy barrier lower than for the monoadduct reaction of Ru(II) complex with ds-DNA. Copyright © 2014 Wiley Periodicals, Inc.

  17. Detection of Campylobacter species and Arcobacter butzleri in stool samples by use of real-time multiplex PCR

    NARCIS (Netherlands)

    R.F. de Boer (Richard); A. Ott (Alewijn); P. Güren (Pinar); E. van Zanten; A.F. van Belkum (Alex); A.M.D. Kooistra-Smid

    2013-01-01

    textabstractThe presence of Campylobacter (or Campylobacter-like) species in stools from patients suspected of infectious gastroenteritis (n = 493) was investigated using real-time PCR for detection of Arcobacter butzleri (hsp60 gene), Campylobacter coli (ceuE gene), Campylobacter jejuni (mapA),

  18. A loop-mediated isothermal amplification (LAMP assay for early detection of Schistosoma mansoni in stool samples: a diagnostic approach in a murine model.

    Directory of Open Access Journals (Sweden)

    Pedro Fernández-Soto

    2014-09-01

    Full Text Available Human schistosomiasis, mainly due to Schistosoma mansoni species, is one of the most prevalent parasitic diseases worldwide. To overcome the drawbacks of classical parasitological and serological methods in detecting S. mansoni infections, especially in acute stage of the disease, development of cost-effective, simple and rapid molecular methods is still needed for the diagnosis of schistosomiasis. A promising approach is the loop-mediated isothermal amplification (LAMP technology. Compared to PCR-based assays, LAMP has the advantages of reaction simplicity, rapidity, specificity, cost-effectiveness and higher amplification efficiency. Additionally, as results can be inspected by the naked eye, the technique has great potential for use in low-income countries.A sequence corresponding to a mitochondrial S. mansoni minisatellite DNA region was selected as a target for designing a LAMP-based method to detect S. mansoni DNA in stool samples. We used a S. mansoni murine model to obtain well defined stool and sera samples from infected mice with S. mansoni cercariae. Samples were taken weekly from week 0 to 8 post-infection and the Kato-Katz and ELISA techniques were used for monitoring the infection. Primer set designed were tested using a commercial reaction mixture for LAMP assay and an in house mixture to compare results. Specificity of LAMP was tested using 16 DNA samples from different parasites, including several Schistosoma species, and no cross-reactions were found. The detection limit of our LAMP assay (SmMIT-LAMP was 1 fg of S. mansoni DNA. When testing stool samples from infected mice the SmMIT-LAMP detected S. mansoni DNA as soon as 1 week post-infection.We have developed, for the first time, a cost-effective, easy to perform, specific and sensitive LAMP assay for early detection of S. mansoni in stool samples. The method is potentially and readily adaptable for field diagnosis and disease surveillance in schistosomiasis-endemic areas.

  19. Intestinal cytomegalovirus disease in immunocompromised patients may be ruled out by search for cytomegalovirus DNA in stool samples.

    OpenAIRE

    Michel, D; Marre, E; Hampl, W; Roczkos, J; Müller, S; Hertenstein, B; Kern, P; Heymer, B; Salzberger, B; Arasteh, K

    1995-01-01

    Cytomegalovirus (CMV) PCR from stool specimens was adopted as a diagnostic tool for patients with suspected CMV colitis. After being established, the method was evaluated in 17 AIDS patients and 19 other immunocompromised patients by comparison of PCR results with clinical, histological, and microbiological or virological data. CMV PCR was positive in 4 symptomatic patients with proven CMV colitis and negative in 15 of 16 patients without characteristic histopathology. Neither CMV immunoglobu...

  20. A SYBR®Green-based real-time PCR method for improved detection of mcr-1-mediated colistin resistance in human stool samples.

    Science.gov (United States)

    Donà, Valentina; Bernasconi, Odette J; Kasraian, Sara; Tinguely, Regula; Endimiani, Andrea

    2017-06-01

    The aim of this study was to design a rapid and sensitive real-time PCR (rt-PCR) method for colistin resistance mcr-1 gene detection in human faecal samples. Stools (n=88) from 36 volunteers were analysed. To isolate mcr-1-producing Enterobacteriaceae, samples were enriched overnight in Luria-Bertani (LB) broth containing 2mg/L colistin and were then plated on selective agar plates with 4mg/L colistin. A SYBR ® Green-based rt-PCR targeting mcr-1 was then designed. For method validation and to establish the limit of detection (LOD), total DNA was extracted from mcr-1-negative and mcr-1-positive Escherichia coli. rt-PCR was also performed with DNA extracted from 88 native stools and after enriching them in LB broth containing colistin. Based on the culture approach, three unique volunteers resulted colonised with mcr-1-harboring E. coli strains. For culture isolates, rt-PCR exhibited a LOD of 10 genomic copies/reaction, with both sensitivity and specificity of 100%. Nevertheless, when testing native stools, only two of the three mcr-1-positive specimens were detected. However, after enrichment in LB broth containing colistin, the rt-PCR was strongly positive for all culture-positive samples. The average cycle threshold was 22, granting rapid and confident detection of positive specimens within 30 cycles. No false positives were observed for the remaining 85 culture-negative specimens. A rapid rt-PCR for detection of mcr-1 from stool specimens was developed. The detection rate was increased by testing selective broth enrichments. This approach also has the advantage of concomitant isolation of mcr-1-harboring strains for further antimicrobial susceptibility and genetic testing. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  1. Stool Test: H. Pylori Antigen

    Science.gov (United States)

    ... Development Infections Diseases & Conditions Pregnancy & Baby Nutrition & Fitness Emotions & Behavior School & Family Life First Aid & Safety Doctors & ... it eradicated the infection. Preparation Unlike most other lab tests, a stool sample is often collected by ...

  2. Evaluation of factors affecting real-time PCR performance for diagnosis of Entamoeba histolytica and Entamoeba dispar in clinical stool samples.

    Science.gov (United States)

    Forsell, Joakim; Koskiniemi, Satu; Hedberg, Ida; Edebro, Helén; Evengård, Birgitta; Granlund, Margareta

    2015-09-01

    Although PCR offers the potential for sensitive detection of parasites there are several pitfalls for optimal performance, especially when DNA is extracted from a complex sample material such as stool. With the aid of a sensitive inhibitor control in a duplex real-time PCR (qPCR) for identification of Entamoeba histolytica and Entamoeba dispar we have evaluated factors that influenced the performance of the qPCR and have suggested a rationale to be used in the analysis of clinical samples. Pre-PCR processing was found to be of outmost importance for an optimal amplification since inhibitors caused false-negative results when higher amounts of sample were used. Stool sampling with a flocked swab (ESwab, Copan), yielding on average 173 mg, gave positive qPCR results in samples with cysts of E. dispar that were negative in serially diluted stool samples. The degree of inhibition found varied between samples and was not an on-off phenomenon. Even low-grade inhibition, shown as an increase of two cycles in the qPCR for the inhibitor control, could lead to false negativity in samples with low amounts of parasites. Lack of amplification in the qPCR due to inhibition could be overcome by dilution of the extracted DNA by 1/10-1/20. We also describe the use of guanidinium thiocyanate buffer for transport and storage of samples as well as a time-saving semi-automated DNA extraction method in an Arrow instrument (Nordiag) preceded by bead beating.

  3. Comparison of methods for detection of Blastocystis infection in routinely submitted stool samples, and also in IBS/IBD Patients in Ankara, Turkey.

    Directory of Open Access Journals (Sweden)

    Funda Dogruman-Al

    Full Text Available BACKGROUND: This study compared diagnostic methods for identifying Blastocystis in stool samples, and evaluated the frequency of detection of Blastocystis in patients with irritable bowel syndrome (IBS and inflammatory bowel disease (IBD. RESULTS AND DISCUSSION: From a set of 105 stool specimens submitted for routine parasitological analysis, 30 were identified as positive for Blastocystis by the culture method. From that group of 30 positives, Lugol's stain, trichrome staining, and an immunofluorescence assay identified 11, 15, and 26 samples as positive respectively. Using culture as a standard, the sensitivity of Lugol's stain was 36.7%, trichrome staining was 50%, and the IFA stain was 86.7%. The specificity of Lugol's stain was 91%, trichrome staining was 100%, and the IFA stain was 97.3%. In the group of 27 IBS and IBD patients, using all methods combined, we detected Blastocystis in 67% (18/27 of the patients. Blastocystis was detected in 33% (2/6 of IBD patients and 76% (16/21 of IBS patients. For comparison, trichrome staining alone, the method most frequently used in many countries, would have only identified Blastocystis infection in 29% (6/21 of the IBS patients. No parasitic co-infections were identified in the IBS/IBD patients. Most Blastocystis-positive IBS/IBD patients were over 36 with an average length of illness of 4.9 years. CONCLUSIONS: Most IBS patients in this study were infected with Blastocystis. IFA staining may be a useful alternative to stool culture, especially if stool specimens have been chemically preserved.

  4. Identification of serotypes and virulence markers of Escherichia coli isolated from human stool and urine samples in Egypt

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    K M Osman

    2012-01-01

    Full Text Available Purpose: Haemorrhagic colitis and haemolytic-uremic syndrome are associated with Shiga-toxin producing Escherichia coli (STEC. There are others DEC (Diarrhoeagenic E. coli pathotypes responsible for outbreaks and others toxins associated to these. Most clinical signs of disease arise as a consequence of the production of Shiga toxin 1 (Stx1, Stx2 or combinations of these toxins. Other major virulence factors include E. coli haemolysin (hlyA, and intimin, the product of the eaeA gene that is involved in the attaching and effacing adherence phenotype. Materials and Methods: In this study, the PCR assay was used to detect 12 E. coli genes associated with virulence (stx1, stx2, hylA, Flic h7 , stb, F41, K99, sta, F17, LT-I, LT-II and eaeA. Results: A total of 108 E. coli strains were serotyped into 64 typable strains. The investigated strains from the stool, 8/80 (10% strains were O 164:K, while the 56/110 strains isolated from the urine were O126:K71 (44/110, 40% and O 86:K 61 (12/110, 11%. The distribution pattern of the detected virulence genes was observed to be in the following order: F17 (10% from the stool and 44% from the urine, Sta (10% from the stool, hylA (10% from the stool and 44% from the urine, Stb (44% from the urine and stx1 (27% from the urine. The 8 faecal strains encoded a combination of the F17, Sta and hylA genes, while the 56 urine strains encoded a combination of the F17 0+ Stb + hylA (44/110, 40% and Stx1 only (12/60, 20%. Conclusion: This is the first report on the molecular characterization of E. coli diarrhoeagenic strains in Egypt and the first report on the potential role of E. coli in diarrhoea and urinary tract infections in a localized geographic area where the people engage in various occupational activities.

  5. Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

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    Roberta Flávia Ribeiro Rolando

    2012-06-01

    Full Text Available This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR. A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

  6. Determination of enterotoxigenic Clostridium perfringens by detecting of the cpa and cpe genes in stool samples of human origin, associated to gastrointestinal disease

    International Nuclear Information System (INIS)

    Oropeza Barrios, Gletty

    2014-01-01

    A molecular methodology is provided to the Centro Nacional de Referencia de Bacteriologia (CNRB) of the Instituto Costarricense de Investigacion y Ensenanza en Nutricion y Salud. An opportune diagnosis is realized of enterotoxigenic Clostridium perfringens in stool samples of sporadic cases and cases associated to foodborne disease outbreaks. DNA extraction of the white microorganism was performed through the methodology implemented in the CNRB. The technique of polymerase chain reaction (PCR) were adapted and standardized to establish the identification of C. perfringens to species level and detection of cpe gene coding for enterotoxin. The sensitivity of the method was determined in a selective culture medium for C. perfringens (Tryptose sulfite cycloserine Agar). A detection limit of about 2,3 x 10 4 CFU/ml was reached for the cpe gene and at least 2,8 x 10 2 CFU/ml for the cpa gene. Retrospective analysis of 61 samples of diarrheal stool suspicious by C. perfringens is performed to evaluate the efficacy of the technique. Three outbreaks caused by C. perfringens were identified and a 10% of positivity in the samples were obtained analyzed during the period between July 2012-March 2014 [es

  7. Differential Diagnosis of Entamoeba spp. in Clinical Stool Samples Using SYBR Green Real-Time Polymerase Chain Reaction

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    Thiago dos Santos Gomes

    2014-01-01

    Full Text Available Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (Tm was 73°C and 70°C, respectively. For E. hartmanni, the Tm was 73°C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries.

  8. Differential diagnosis of Entamoeba spp. in clinical stool samples using SYBR green real-time polymerase chain reaction.

    Science.gov (United States)

    Gomes, Thiago Dos Santos; Garcia, Mariana Coimbra; de Souza Cunha, Flavia; Werneck de Macedo, Heloisa; Peralta, José Mauro; Peralta, Regina Helena Saramago

    2014-01-01

    Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (T(m)) was 73 °C and 70 °C, respectively. For E. hartmanni, the T(m) was 73 °C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries.

  9. Differential Diagnosis of Entamoeba spp. in Clinical Stool Samples Using SYBR Green Real-Time Polymerase Chain Reaction

    Science.gov (United States)

    Gomes, Thiago dos Santos; Garcia, Mariana Coimbra; de Souza Cunha, Flavia; Peralta, José Mauro; Peralta, Regina Helena Saramago

    2014-01-01

    Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (T m) was 73°C and 70°C, respectively. For E. hartmanni, the T m was 73°C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries. PMID:24693242

  10. The origin of endodontic Enterococcus faecalis explored by comparison of virulence factor patterns and antibiotic resistance to that of isolates from stool samples, blood cultures and food.

    Science.gov (United States)

    Vidana, R; Rashid, M U; Özenci, V; Weintraub, A; Lund, B

    2016-04-01

    To elucidate the origin of Enterococcus faecalis isolated from secondary root canal infections and the possibility for a foodborne transmission by comparing them to strains recovered from food, blood and stool regarding putative virulence factors and antibiotic susceptibility profiles, where strains from common origin were hypothesized to harbour similar characteristics. A total of 108 E. faecalis strains recovered in the county of Stockholm, Sweden, were screened using PCR for putative virulence factors esp, cylA, gelE/gelatinase-negative phenotype (ef1841/fsrC), efaA, ace and asa1. The minimum inhibitory concentration (MIC) for ampicillin, piperacillin-tazobactam, imipenem, gentamicin, vancomycin, ciprofloxacin and linezolid was determined using the agar dilution method. Next to strains from blood, the food isolates presented the highest average number of virulence determinants and were frequently enriched with asa1 coding for aggregation substance. None of the endodontic strains carried cylA, and the gelatinase-negative phenotype caused by a deletion dominated the group. Altogether, the most prevalent genes were gelE, efaA and ace, and a combination of them was equally present in approximately 80% of the strains from food, stool and root canals in comparison with 43.3% of the blood isolates. High-level resistance to ciprofloxacin and gentamicin was observed in 30% of the blood isolates, whereas the isolates from other origins, with single exceptions, were susceptible to all tested antibiotics. Evidence for a foodborne transmission, explaining the high reported prevalence of E. faecalis in root filled teeth, could not be determined based on the similarities in virulence factor patterns and antibiotic susceptibility. The only linkage between isolates from food and root canals consisted of a shared common combination of the genes gelE, efaA and ace. The high occurrence of putative virulence traits in food isolates questions the safety of E. faecalis in food

  11. Detection and species identification of Campylobacter in stool samples of children and animals from Vellore, south India

    Directory of Open Access Journals (Sweden)

    P Rajendran

    2012-01-01

    Full Text Available Campylobacter spp. are an important cause of bacterial gastroenteritis frequently isolated from animal, poultry and environmental samples. In this study, we investigated the zoonotic potential of Campylobacter spp. by comparing prevalence rates and species in 394 children with diarrhoea and 652 animals in Vellore using PCR-based tools. Eighteen children (4.5% had campylobacteriosis, a majority of whom had co-pathogens (15/18 and most were infected with Campylobacter jejuni (16/18. A few C. coli and mixed infections with both species were also seen. Among the animal samples, 16/25 chicken samples (64% were positive and all were found to be C. jejuni.

  12. Study of the Prevalence of Causative Bacterial&Protozoal Agents of in Stool Samples of 470 Gastroenteritis Patients Referring to the Nikoopour Clinic in Yazd,Iran

    Directory of Open Access Journals (Sweden)

    MR Sharifi

    2004-04-01

    Full Text Available Interoduction: Gasteroenteritis is one of the problems worth consideration all over the world. It is one of the important causes of mortality, especially in children < 5 years of age, in developing countries including Iran. The aim of this descriptive study was to determine the demographic conditions influencing the presence of causative bacteria and protozoa, followed by antibiograms of isolated bacteria from stool samples of patients with gasteroenteritis referring to Nikoopour Clinic in the city of Yazd, Iran from 1998 – 2001. Materials and method: A total of 470 samples were microbiologically examined by direct method, culture and then antibiogramed. In order to isolate the possible bacteria, differential and selected media were used. Also, wet – mount technique was applied for detection of protozoa. Results: Results revealed that 272 samples (57.9% were infected by pathogenic bacteria or protozoa. 138 (50.8% pathogenic specimens were from male patients and the remaining 134(49.3% were from female patients. Isolated species were: Enteropathogenic E.coli 117(43%, Shigella 51(18.8%, Salmonella.interetidis 25(9.2%, C.jejuni 16(5.9%, Giardia lambdia 51(18.8% and Amoebae spp 12(4.4%. The most commonly detected shigella species was dysenteriae, (74.5% while boydii with 2% was the least common type observed in the specimens. Except shigella, all the other bacteria were more common in males than female, but insignificant statistically. In order to determine the sensitivity and/or resistance of pathogenic bacteria, antibiogram test was performed using selected antibiotic disks such as Ampicillin, Nalidixic Acid, Ciprofloxacin, Gentamycin and Sulfamethaxazole. Conclusion: Results revealed that some patients were probably infected by pathogenic factors other than bacteria or protozoa. Since all viruses and parasites are almost resistant to antibiotics and on the other hand, administration of antibiotics may lead to resistance of bacterial agents

  13. Microscopic diagnosis of sodium acetate-acetic acid-formalin-fixed stool samples for helminths and intestinal protozoa: a comparison among European reference laboratories.

    Science.gov (United States)

    Utzinger, J; Botero-Kleiven, S; Castelli, F; Chiodini, P L; Edwards, H; Köhler, N; Gulletta, M; Lebbad, M; Manser, M; Matthys, B; N'Goran, E K; Tannich, E; Vounatsou, P; Marti, H

    2010-03-01

    The present study aimed to compare the diagnostic performance of different European reference laboratories in diagnosing helminths and intestinal protozoa, using an ether-concentration method applied to sodium acetate-acetic acid-formalin (SAF)-preserved faecal samples. In total, 102 stool specimens were analysed during a cross-sectional parasitological survey in urban farming communities in Côte d'Ivoire. Five SAF-preserved faecal samples were prepared from each specimen and forwarded to the participating reference laboratories, processed and examined under a microscope adhering to a standard operating procedure (SOP). Schistosoma mansoni (cumulative prevalence: 51.0%) and hookworm (cumulative prevalence: 39.2%) were the predominant helminths. There was excellent agreement (kappa > 0.8; p protozoa were Entamoeba coli (median prevalence: 67.6%), Blastocystis hominis (median prevalence: 55.9%) and Entamoeba histolytica/Entamoeba dispar (median prevalence: 47.1%). Substantial agreement among reference laboratories was found for E. coli (kappa = 0.69), but only fair or moderate agreement was found for other Entamoeba species, Giardia intestinalis and Chilomastix mesnili. There was only poor agreement for B. hominis, Isospora belli and Trichomonas intestinalis. In conclusion, although common helminths were reliably diagnosed by European reference laboratories, there was only moderate agreement between centres for pathogenic intestinal protozoa. Continued external quality assessment and the establishment of a formal network of reference laboratories is necessary to further enhance both accuracy and uniformity in parasite diagnosis.

  14. Stool Color: When to Worry

    Science.gov (United States)

    Stool color: When to worry Yesterday, my stool color was bright green. Should I be concerned? Answers from Michael ... M.D. Stool comes in a range of colors. All shades of brown and even green are ...

  15. Sediminibacillus massiliensis sp. nov., a moderately halophilic, Gram-positive bacterium isolated from a stool sample of a young Senegalese man.

    Science.gov (United States)

    Senghor, Bruno; Bassène, Hubert; Khelaifia, Saber; Robert, Catherine; Fournier, Pierre-Edouard; Ruimy, Raymond; Sokhna, Cheikh; Raoult, Didier; Lagier, Jean-Christophe

    2018-02-07

    A Gram-positive, moderately halophilic bacterium, referred to as strain Marseille-P3518 T , was isolated from a stool sample with 2% NaCl concentration from a healthy 15-year-old male living in Dielmo, a village in Senegal. Cells are aerobic, rod-shaped and motile and display endospore formation. Strain Marseille-P3518 T can grow in a medium with 0-20% (w/v) sodium chloride (optimally at 5-7.5% w/v). The major fatty acids were 12-methyl-tetradecanoic acid (45.8%), 13-methyl-tetradecanoic acid (26.9%) and 12-methyl-tridecanoic acid (12.8%). The genome is 4,347,479 bp long with 42.1% G+C content. It contains 4282 protein-coding and 107 RNA genes. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain Marseille-P3518 T is a member of the Bacillaceae family and is closely related to Sediminibacillus albus (97.4% gene sequence similarity). Strain Marseille-P3518 T was clearly differentiated from its phylogenetic neighbors on the basis of phenotypic and genotypic features. Strain Marseille-P3518 T is, therefore, considered to be a novel representative of the genus Sediminibacillus, for which the name Sediminibacillus massiliensis sp. nov. is proposed, and the type strain is Marseille-P3518 T (CSUR P3518T, DSM69894).

  16. Cases of cryptosporidiosis co-infections in AIDS patients: a correlation between clinical presentation and GP60 subgenotype lineages from aged formalin-fixed stool samples.

    Science.gov (United States)

    Del Chierico, F; Onori, M; Di Bella, S; Bordi, E; Petrosillo, N; Menichella, D; Cacciò, S M; Callea, F; Putignani, L

    2011-07-01

    Nine cases of cryptosporidiosis co-infections in AIDS patients were clinically categorised into severe (patients 1, 3, 8 and 9), moderate (patients 4 and 5) and mild (patients 2, 6 and 7). Formalin-fixed faecal specimens from these patients were treated to obtain high quality DNA competent for amplification and sequencing of the 60-kDa glycoprotein (GP60) gene. Sequence analysis revealed that one patient was infected with Cryptosporidium hominis whereas the remaining eight patients were infected with C. parvum. Interestingly, the patients showing severe cryptosporidiosis harboured two subtypes within the C. parvum allelic family IIc (IIcA5G3 and IIcA5G3R2), whereas patients with moderate or mild infections showed various subtypes of the C. parvum allelic family IIa (IIaA14G2R1, IIaA15G2R1, IIaA17G3R1 and IIaA18G3R1). DNA extraction and genotyping of Cryptosporidium spp. is a challenging task on formalin-fixed stool samples, whose diagnostic outcome is age-dependent. The method herein reported represents a step forward routine diagnosis and improves epidemiology of HIV-related clinical cases. Due to the need to elucidate genetic richness of Cryptosporidium human isolates, this approach represents a useful tool to correlate individual differences in symptoms to subgenotyping lineages.

  17. Stool Gram stain

    Science.gov (United States)

    ... of stool; Feces Gram stain References Allos BM. Campylobacter infections. In: Goldman L, Schafer AI, eds. Goldman- ... Bacterial Infections Read more Foodborne Illness Read more Gastroenteritis Read more A.D.A.M., Inc. is ...

  18. Novel real-time PCr for detection of Schistosoma japonicum in stool.

    Science.gov (United States)

    Lier, T; Simonsen, G S; Haaheim, H; Hjelmevoll, S O; Vennervald, B J; Johansen, M V

    2006-03-01

    Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

  19. Probiotic properties of lactic acid bacteria isolated from stool samples of longevous people in regions of Hotan, Xinjiang and Bama, Guangxi, China.

    Science.gov (United States)

    Gu, Rui-Xia; Yang, Zhen-Quan; Li, Zheng-Hua; Chen, Shun-Li; Luo, Zhen-Lan

    2008-12-01

    A total of 567 lactic acid bacteria (LAB) strains were isolated from the stool samples of longevous people in regions of Hotan, Xinjiang and Bama, Guangxi, China. In order to reduce the number of strains for further examinations, 36 isolates were screened out for further examination whilst the other strains, which had lower probiotic properties, were not suitable for yogurt production due to the absence of growth in pH 3.5 MRS medium and no curding during fermentation, and so were excluded. The result of identification by API, sequence analysis of 16S rRNA and random amplified polymorphic DNA (RAPD) analysis showed that there were three strains of Lactobacillus acidophilus, 10 strains of Lactobacillus rhamnosus, three strains of Lactobacillus casein, three strains of Lactobacillus brevis, two strains of Enterococcus faecium, two strains of Enterococcus faecalis, four strains of Bifdibacterium infantis, three strains of Bifdibacterium brevise, three strains of Bifdibacterium bifidium, two strains of Bifdibacterium adolecentis and one strain of Bifdibacterium longam among the 36 isolates. These strains were evaluated by in vitro methods including survival upon exposure to pH 2.0, 3.0 and/or 0.3% oxgall and adhesion to the human colon adenocarcinoma cell line Caco-2 as well as antimicrobial activity against potential pathogens. The results presented here show that L. rhamnosus LV108, L. rhamnosus F, B. brevise R39 and B. infantis R42 are acid and bile tolerant, adhere to the cultured human intestinal Caco-2 cell line, antagonistic activity against potential pathogenic bacteria infection in vitro, and so are potential strains for probiotic use.

  20. Lack of Correlation between Bristol Stool Scale and Quantitative Bacterial Load in Infection

    OpenAIRE

    Abrar K. Thabit; David P. Nicolau

    2015-01-01

    Decision to test for Clostridium difficile infection (CDI) is usually made when patients have loose stools with Bristol stool score of ≤5. We aimed to assess the relationship between bacterial load of C. difficile and Bristol stool scale, as well as stool frequency in stool samples collected from patients infected with the organism. Samples were collected at baseline, during therapy, and at the end of therapy. Spearman correlation test was used to evaluate these relationships. No correlation ...

  1. Comparison of the Triage Micro Parasite Panel and Microscopy for the Detection of Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Cryptosporidium parvum in Stool Samples Collected in Kenya

    Directory of Open Access Journals (Sweden)

    Brett Swierczewski

    2012-01-01

    Full Text Available Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are three of the most important parasitic causes of acute diarrhea worldwide. Laboratory diagnosis of these parasites is usually done by ova and parasite examination (O&P examination via microscopy. The sensitivity and specificity of O&P examination varies among laboratories and can be labor intensive and time consuming. The Triage Micro Parasite Panel (BioSite, San Diego, California is an enzyme immunoassay kit that can detect E. histolytica/E. dispar, G. lamblia, and C. parvum simultaneously using fresh or frozen stool. The present study evaluated the Triage Micro Parasite Panel in detecting E. histolytica/E. dispar, G. lamblia, and C. parvum compared to O&P examination in 266 stool samples collected at medical facilities in Kenya. The sensitivity and specificity results for the Triage Micro Parasite Panel were: for E. histolytica/E. dispar: 100%, 100%, G. lamblia: 100%, 100% and C. parvum: 73%, 100%. There was no evidence of cross reactivity using the kit with other parasites identified in the stool specimens. These results indicate that the Triage Micro Parasite Panel is a highly sensitive kit that can be used for screening purposes in large scale studies or outbreak investigations or as a possible alternative to O&P examination.

  2. First detection and genotyping of Giardia intestinalis in stool samples collected from children in Ghazni Province, eastern Afghanistan and evaluation of the PCR assay in formalin-fixed specimens.

    Science.gov (United States)

    Lass, Anna; Karanis, Panagiotis; Korzeniewski, Krzysztof

    2017-08-01

    It is estimated that faecal-orally transmitted diseases are common in Afghanistan, as a consequence of poor hygienic standards of life and widespread contamination of water and food with both human and animal faeces. However, there is little information in the literature concerning infections caused by intestinal parasites in the Afghan population. In this study, we report the occurrence of Giardia intestinalis assemblages (A and B) in formalin-fixed stool samples collected from 245 Afghan schoolchildren living in Ghazni Province in eastern Afghanistan. Detection of the parasite's DNA and genotyping was performed using real-time PCR, specific to the β-giardin gene of G. intestinalis. Positive results were recorded in 52 (21.2%) samples. Genotyping was successful in 39 faecal samples and showed the predominance of assemblage B of G. intestinalis in this population (15 assemblage A and 24 assemblage B). Co-infection with both genotypes A and B was detected in four samples. Additionally, we evaluated the effect of 10% buffered formalin fixative on the detection of G. intestinalis DNA using real-time PCR and nested PCR characterised by different lengths of PCR products (74 and 479 bp, respectively). The human faeces containing the Giardia cysts were tested for 16 weeks. Amplification of G. intestinalis DNA with real-time PCR was possible up to 6 weeks of preservation of stool sample in formalin, compared to only 2 weeks with nested PCR. This suggests that real-time PCR is a more suitable tool in cases where stool samples have to be kept in formalin for longer periods of time.

  3. One-step multiplex real-time RT-PCR assay for detecting and genotyping wild-type group A rotavirus strains and vaccine strains (Rotarix® and RotaTeq®) in stool samples

    Science.gov (United States)

    Mijatovic-Rustempasic, Slavica; Esona, Mathew D.; Tam, Ka Ian; Quaye, Osbourne; Bowen, Michael D.

    2016-01-01

    Background. Group A rotavirus (RVA) infection is the major cause of acute gastroenteritis (AGE) in young children worldwide. Introduction of two live-attenuated rotavirus vaccines, RotaTeq® and Rotarix®, has dramatically reduced RVA associated AGE and mortality in developed as well as in many developing countries. High-throughput methods are needed to genotype rotavirus wild-type strains and to identify vaccine strains in stool samples. Quantitative RT-PCR assays (qRT-PCR) offer several advantages including increased sensitivity, higher throughput, and faster turnaround time. Methods. In this study, a one-step multiplex qRT-PCR assay was developed to detect and genotype wild-type strains and vaccine (Rotarix® and RotaTeq®) rotavirus strains along with an internal processing control (Xeno or MS2 RNA). Real-time RT-PCR assays were designed for VP7 (G1, G2, G3, G4, G9, G12) and VP4 (P[4], P[6] and P[8]) genotypes. The multiplex qRT-PCR assay also included previously published NSP3 qRT-PCR for rotavirus detection and Rotarix® NSP2 and RotaTeq® VP6 qRT-PCRs for detection of Rotarix® and RotaTeq® vaccine strains respectively. The multiplex qRT-PCR assay was validated using 853 sequence confirmed stool samples and 24 lab cultured strains of different rotavirus genotypes. By using thermostable rTth polymerase enzyme, dsRNA denaturation, reverse transcription (RT) and amplification (PCR) steps were performed in single tube by uninterrupted thermocycling profile to reduce chances of sample cross contamination and for rapid generation of results. For quantification, standard curves were generated using dsRNA transcripts derived from RVA gene segments. Results. The VP7 qRT-PCRs exhibited 98.8–100% sensitivity, 99.7–100% specificity, 85–95% efficiency and a limit of detection of 4–60 copies per singleplex reaction. The VP7 qRT-PCRs exhibited 81–92% efficiency and limit of detection of 150–600 copies in multiplex reactions. The VP4 qRT-PCRs exhibited 98.8

  4. Ultrasensitive Detection of Shigella Species in Blood and Stool.

    Science.gov (United States)

    Luo, Jieling; Wang, Jiapeng; Mathew, Anup S; Yau, Siu-Tung

    2016-02-16

    A modified immunosensing system with voltage-controlled signal amplification was used to detect Shigella in stool and blood matrixes at the single-digit CFU level. Inactivated Shigella was spiked in these matrixes and detected directly. The detection was completed in 78 min. Detection limits of 21 CFU/mL and 18 CFU/mL were achieved in stool and blood, respectively, corresponding to 2-7 CFUs immobilized on the detecting electrode. The outcome of the detection of extremely low bacterium concentration, i.e., below 100 CFU/mL, blood samples show a random nature. An analysis of the detection probabilities indicates the correlation between the sample volume and the success of detection and suggests that sample volume is critical for ultrasensitive detection of bacteria. The calculated detection limit is qualitatively in agreement with the empirically determined detection limit. The demonstrated ultrasensitive detection of Shigella on the single-digit CFU level suggests the feasibility of the direct detection of the bacterium in the samples without performing a culture.

  5. Acceptability of study procedures (self-collected introital swabs, blood draws and stool sample collection) by students 10-16 years for an HPV vaccine effectiveness study: a pilot study.

    Science.gov (United States)

    Nakalembe, Miriam; Mutyaba, Twaha; Mirembe, Florence

    2016-03-16

    A cohort study was planned to evaluate vaccine immunogenicity and effect of malaria and helminth co-infections on the bivalent Human papilloma virus (HPV) vaccine. The study would involve self collected introital swabs, blood draws and stool sample collection. We therefore conducted a pilot study to assess the acceptability of these procedures among the students and their parents. A cross-sectional study among forty four students from two purposively selected primary schools of Western Uganda. Exit interviews and two focus group discussions (FGD) (for parents) were conducted. Acceptability was measured by willingness to undergo the procedures again, recommending the procedures to others as well as proportion of introital swabs positive for β globulin. FGD determined acceptability of the parents and explored opinions and perceptions that would influence their decisions. HPV-16/18 and β globulin deoxyribonucleic acid (DNA) were analysed using a polymerase chain reaction (PCR) kit. All the students (100%) in the study were willing to provide a self- collected introital swab and a stool sample as well as recommending their friends while (86.3%) were willing for blood draws. There were 40/44 (90.1%) self collected introital swabs that had positive result for human β globulin though none of them was positive for HPV-16/18. In the FGD, it emerged that parents concerns were on the blood draws and introital swab collection which were addressed. The study procedures were highly acceptable among this study population of students and their parents. Follow-up to assess HPV vaccine effectiveness and factors that may influence the vaccine in this age group is feasible.

  6. Flushable reagent stool blood test

    Science.gov (United States)

    Stool occult blood test - flushable home test; Fecal occult blood test - flushable home test ... gastrointestinal tract, include: Coughing up and then swallowing blood Nose bleed Abnormal test results require follow-up with your doctor.

  7. a comparison between the use of single and composite samples

    African Journals Online (AJOL)

    Dr F Neser

    A comparison between single and composite milk samples for the genetic evaluation of milk composition in dairy cattle ... Dairy Cattle Performance Testing Scheme replaced composite sampling with a single sample in 1995. The implementation of the new Scheme was ... Materials and methods. To obtain information on the ...

  8. The Bristol Stool Scale and Its Relationship to Clostridium difficile Infection

    OpenAIRE

    Caroff, Daniel A.; Edelstein, Paul H.; Hamilton, Keith; Pegues, David A.

    2014-01-01

    The Bristol stool form scale classifies the relative density of stool samples. In a prospective cohort study, we investigated the associations between stool density, C. difficile assay positivity, hospital-onset C. difficile infection, complications, and severity of C. difficile. We describe associations between the Bristol score, assay positivity, and clinical C. difficile infection.

  9. The Bristol stool scale and its relationship to Clostridium difficile infection.

    Science.gov (United States)

    Caroff, Daniel A; Edelstein, Paul H; Hamilton, Keith; Pegues, David A

    2014-09-01

    The Bristol stool form scale classifies the relative density of stool samples. In a prospective cohort study, we investigated the associations between stool density, C. difficile assay positivity, hospital-onset C. difficile infection, complications, and severity of C. difficile. We describe associations between the Bristol score, assay positivity, and clinical C. difficile infection. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  10. Isolation of Campylobacter Species from Stool Samples by Use of a Filtration Method: Assessment from a United States-Based Population.

    Science.gov (United States)

    Nachamkin, Irving; Nguyen, Phi

    2017-07-01

    Fecal samples submitted to our clinical microbiology laboratory from patients in the Philadelphia region were prospectively analyzed for Campylobacter species other than C. jejuni and C. coli using a filtration method and microaerobic conditions with increased H 2 concentrations. Of 225 samples tested, 13 (5.8%) yielded Campylobacter species, with frequent isolation of C. concisus The majority of Campylobacter species were not clinically significant. Additional studies in U.S. populations are warranted. Copyright © 2017 American Society for Microbiology.

  11. 21 CFR 868.6700 - Anesthesia stool.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Anesthesia stool. 868.6700 Section 868.6700 Food... DEVICES ANESTHESIOLOGY DEVICES Miscellaneous § 868.6700 Anesthesia stool. (a) Identification. An anesthesia stool is a device intended for use as a stool for the anesthesiologist in the operating room. (b...

  12. A single-sampling hair trap for mesocarnivores

    Science.gov (United States)

    Jonathan N. Pauli; Matthew B. Hamilton; Edward B. Crain; Steven W. Buskirk

    2007-01-01

    Although techniques to analyze and quantifY DNA-based data have progressed, methods to noninvasively collect samples lag behind. Samples are generally collected from devices that permit coincident sampling of multiple individuals. Because of cross-contamination, substantive genotyping errors can arise. We developed a cost-effective (US$4.60/trap) single-capture hair...

  13. Multiplex Real-Time PCR Assay Targeting Eight Parasites Customized to the Korean Population: Potential Use for Detection in Diarrheal Stool Samples from Gastroenteritis Patients.

    Directory of Open Access Journals (Sweden)

    Eun Jeong Won

    Full Text Available Intestinal parasitic diseases occur worldwide and can cause diarrhea or gastroenteritis; however, their diagnosis is quite difficult, especially in low-endemism countries. We developed a multiplex real-time PCR assay for detection of eight intestinal parasites and prospectively evaluated it for patients with gastroenteritis. The assay targeted Cryptosporidium parvum, Giardia lamblia, Entamoeba histolytica, Blastocystis hominis, Dientamoeba fragilis, Clonorchis sinensis, Metagonimus yokogawai, and Gymnophalloides seoi. Performance characteristics were evaluated based on recovery after DNA extraction, analytical sensitivity, specificity, reproducibility, cross-reactivity, and interference characteristics. Clinical performance was validated against microscopy on 123 diarrheal samples. The assay demonstrated strong correlations between DNA concentrations and Ct values (R2, 0.9924-0.9998, and had a high PCR efficiency (83.3%-109.5%. Polymerase chain reactions detected as few as 10-30 copies of genomic DNA, and coefficient of variance was 0-7%. There was no cross-reactivity to the other 54 microorganisms tested. Interference occurred only in presence of high concentrations of erythrocytes or leukocytes. This assay had a higher correct identification rate (100.0% vs. 90.2% and lower incorrect ID rate (0.0% vs. 9.8% when compared to microscopy. Overall, this assay showed a higher sensitivity (100.0%; 95% confidence interval [CI] of 80.5-100.0 than microscopy (29.4%; 95% CI 10.31-55.96, and the specificity levels were comparable for both methods (100.0%; 95% CI 96.58-100.0. This newly developed multiplex real-time PCR assay offers a potential use for detecting intestinal parasitic pathogens customized to the Korean population.

  14. Development of a thermostabilized, one-step, nested, tetraplex PCR assay for simultaneous identification and differentiation of Entamoeba species, Entamoeba histolytica and Entamoeba dispar from stool samples.

    Science.gov (United States)

    Foo, Phiaw Chong; Chan, Yean Yean; See Too, Wei Cun; Tan, Zi Ning; Wong, Weng Kin; Lalitha, Pattabhiraman; Lim, Boon Huat

    2012-09-01

    Entamoeba histolytica is the only Entamoeba species that causes amoebiasis in humans. Approximately 50 million people are infected, with 100, 000 deaths annually in endemic countries. Molecular diagnosis of Entamoeba histolytica is important to differentiate it from the morphologically identical Entamoeba dispar to avoid unnecessary medication. Conventional molecular diagnostic tests require trained personnel, cold-chain transportation and/or are storage-dependent, which make them user-unfriendly. The aim of this study was to develop a thermostabilized, one-step, nested, tetraplex PCR assay for the detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba species in cold-chain-free and ready-to-use form. The PCR test was designed based on the Entamoeba small subunit rRNA (SSU-rRNA) gene, which detects the presence of any Entamoeba species, and simultaneously can be used to differentiate Entamoeba histolytica from Entamoeba dispar. In addition, a pair of primers was designed to serve as an internal amplification control to help identify inhibitors in the samples. All PCR reagents together with the designed primers were thermostabilized by lyophilization and were stable at 24 °C for at least 6 months. The limit of detection of the tetraplex PCR was found to be 39 pg DNA or 1000 cells for Entamoeba histolytica and 78 pg DNA or 1000 cells for Entamoeba dispar, and the specificity was 100 %. In conclusion, this cold-chain-free, thermostabilized, one-step, nested, multiplex PCR assay was found to be efficacious in differentiating Entamoeba histolytica from other non-pathogenic Entamoeba species.

  15. Performance evaluation of direct saline stool microscopy, Formol ether concentration and Kato Katz diagnostic methods for intestinal parasitosis in the absence of gold standard methods.

    Science.gov (United States)

    Hailu, Tadesse; Abera, Bayeh

    2015-07-01

    The parasite load within the sample and the amount of sample taken during examination greatly compromise the sensitivity of direct saline stool microscopy. A cross-sectional study was conducted in March 2011 in Bahir Dar city among 778 fresh single stool samples to evaluate the performance of direct saline (DS), Kato Katz (KK) and Formol ether concentration (FEC) methods against the 'Gold' standard. Among 778 stool samples from school age children, the highest prevalence of intestinal parasites was recorded by FEC (55.1%). The sensitivity of DS, FEC and KK were 61.1%, 92.3% and 58.7%, respectively. FEC is more sensitive than DS and KK. Hence, use of the latter is preferred. © The Author(s) 2015.

  16. Stools - pale or clay-colored

    Science.gov (United States)

    ... gov/ency/article/003129.htm Stools - pale or clay-colored To use the sharing features on this page, please enable JavaScript. Stools that are pale, clay, or putty-colored may be due to problems ...

  17. Rapid diagnosis of diarrhea caused by Shigella sonnei using dipsticks; comparison of rectal swabs, direct stool and stool culture.

    Directory of Open Access Journals (Sweden)

    Claudia Duran

    Full Text Available BACKGROUND: We evaluated a dipstick test for rapid detection of Shigella sonnei on bacterial colonies, directly on stools and from rectal swabs because in actual field situations, most pathologic specimens for diagnosis correspond to stool samples or rectal swabs. METHODOLOGY/PRINCIPAL FINDINGS: The test is based on the detection of S. sonnei lipopolysaccharide (LPS O-side chains using phase I-specific monoclonal antibodies coupled to gold particles, and displayed on a one-step immunochromatographic dipstick. A concentration as low as 5 ng/ml of LPS was detected in distilled water and in reconstituted stools in 6 minutes. This is the optimal time for lecture to avoid errors of interpretation. In distilled water and in reconstituted stools, an unequivocal positive reaction was obtained with 4 x 10(6 CFU/ml of S. sonnei. The specificity was 100% when tested with a battery of Shigella and different unrelated strains. When tested on 342 rectal swabs in Chile, specificity (281/295 was 95.3% (95% CI: 92.9% - 97.7% and sensitivity (47/47 was 100%. Stool cultures and the immunochromatographic test showed concordant results in 95.5 % of cases (328/342 in comparative studies. Positive and negative predictive values were 77% (95% CI: 65% - 86.5% and 100% respectively. When tested on 219 stools in Chile, Vietnam, India and France, specificity (190/198 was 96% (95% CI 92%-98% and sensitivity (21/21 was 100%. Stool cultures and the immunochromatographic test showed concordant results in 96.3 % of cases (211/219 in comparative studies. Positive and negative predictive values were 72.4% (95% CI 56.1%-88.6% and 100 %, respectively. CONCLUSION: This one-step dipstick test performed well for diagnosis of S. sonnei both on stools and on rectal swabs. These data confirm a preliminary study done in Chile.

  18. Single versus duplicate blood samples in ACTH stimulated adrenal vein sampling

    NARCIS (Netherlands)

    Dekkers, T.; Arntz, M.; Wilt, G.J. van der; Schultze Kool, L.J.; Sweep, F.C.; Hermus, A.R.M.M.; Lenders, J.W.M.; Deinum, J.

    2013-01-01

    BACKGROUND: Adrenal vein sampling (AVS) is the preferred test for subtyping primary aldosteronism. However, the procedure is technically demanding and costly. In AVS it is common practice to take duplicate blood samples at each location. In this paper we explore whether a single sample procedure

  19. Lack of Correlation between Bristol Stool Scale and Quantitative Bacterial Load in Infection

    Directory of Open Access Journals (Sweden)

    Abrar K. Thabit

    2015-01-01

    Full Text Available Decision to test for Clostridium difficile infection (CDI is usually made when patients have loose stools with Bristol stool score of ≤5. We aimed to assess the relationship between bacterial load of C. difficile and Bristol stool scale, as well as stool frequency in stool samples collected from patients infected with the organism. Samples were collected at baseline, during therapy, and at the end of therapy. Spearman correlation test was used to evaluate these relationships. No correlation between Bristol stool scale and fecal load of C. difficile was found for both spores and vegetative cells at all time points as counts were persistently high ( P = non-significant. Weak positive correlations were found between stool frequency and fecal load of C. difficile spores and vegetative cells ( r s = 0.22 and 0.24, P = 0.04 and 0.03, respectively. These findings indicate that quantitative colony counts were sufficiently high to detect C. difficile , irrespective of stool consistency, and suggest that semiformed stool should be sought for the pathogen in symptomatic patients with frequent stools.

  20. Lack of Correlation between Bristol Stool Scale and Quantitative Bacterial Load in Clostridium difficile Infection.

    Science.gov (United States)

    Thabit, Abrar K; Nicolau, David P

    2015-01-01

    Decision to test for Clostridium difficile infection (CDI) is usually made when patients have loose stools with Bristol stool score of ≥5. We aimed to assess the relationship between bacterial load of C. difficile and Bristol stool scale, as well as stool frequency in stool samples collected from patients infected with the organism. Samples were collected at baseline, during therapy, and at the end of therapy. Spearman correlation test was used to evaluate these relationships. No correlation between Bristol stool scale and fecal load of C. difficile was found for both spores and vegetative cells at all time points as counts were persistently high (P = non-significant). Weak positive correlations were found between stool frequency and fecal load of C. difficile spores and vegetative cells (r s = 0.22 and 0.24, P = 0.04 and 0.03, respectively). These findings indicate that quantitative colony counts were sufficiently high to detect C. difficile, irrespective of stool consistency, and suggest that semiformed stool should be sought for the pathogen in symptomatic patients with frequent stools.

  1. Creation and initial evaluation of a Stool Form Scale for children.

    Science.gov (United States)

    Chumpitazi, Bruno P; Lane, Mariella M; Czyzewski, Danita I; Weidler, Erica M; Swank, Paul R; Shulman, Robert J

    2010-10-01

    To develop a pediatric stool form rating scale and determine its interrater reliability, intrarater reliability, and agreement among pediatric gastroenterologists. An ordinal stool scale with 5 categorical stool form types was created on the basis of the Bristol Stool Form Scale, and 32 color 2-dimensional stool photographs were shown to 14 pediatric gastroenterologists. Each gastroenterologist rated the stool form depicted in each photograph with the modified stool scale. Ten gastroenterologists agreed to rerate the stool form depicted in each photograph a minimum of 6 months after the first rating. A total of 448 ratings were completed; 430 (94%) of all ratings were within at least 1 category type of the most common (modal) rating for each photograph. Eight (25%) stool photographs had complete agreement among all raters. Interrater and intrarater reliability was high with a single measure intraclass correlation of 0.85 (95% confidence interval: 0.78-0.91; PBristol Stool Form Scale provided a high degree of interrater reliability, intrarater reliability, and agreement among pediatric gastroenterologists. Copyright (c) 2010 Mosby, Inc. All rights reserved.

  2. Isolation and Identification of Adenovirus Recovered from the Stool ...

    African Journals Online (AJOL)

    In order to establish the role of adenovirus in gastroenteritis in Nigerian children, stool samples were collected from 138 young children with gastroenteritis and 29 other age-matched controls. The samples were inoculated into 6 different tissue culture cell lines and isolates with characteristic CPE were subjected to CFT ...

  3. Identification and antibiotic sensitivity test of bacteria from stools of ...

    African Journals Online (AJOL)

    One hundred and fifty stool samples from 65 female and 85 male patients with acute diarrhoea from the Central Hospital, Agbor (Nigeria) were examined to ascertain the likelihood of cholera outbreak in Agbor. The samples were preserved in Carey-Blair semi-solid medium, inoculated directly on blood agar, McConkey agar ...

  4. a comparison between the use of single and composite samples

    African Journals Online (AJOL)

    Dr F Neser

    South African Society for Animal Science. 44. A comparison between single and composite milk samples for the genetic evaluation of milk composition in dairy cattle. R. van Dyk1, F.W.C. Neser1# & F.H. Kanfer2. 1 Department of Animal Science, University of the Free State, PO Box 339, Bloemfontein 9300, South Africa.

  5. Genomic Sequencing of Single Microbial Cells from Environmental Samples

    Energy Technology Data Exchange (ETDEWEB)

    Ishoey, Thomas; Woyke, Tanja; Stepanauskas, Ramunas; Novotny, Mark; Lasken, Roger S.

    2008-02-01

    Recently developed techniques allow genomic DNA sequencing from single microbial cells [Lasken RS: Single-cell genomic sequencing using multiple displacement amplification, Curr Opin Microbiol 2007, 10:510-516]. Here, we focus on research strategies for putting these methods into practice in the laboratory setting. An immediate consequence of single-cell sequencing is that it provides an alternative to culturing organisms as a prerequisite for genomic sequencing. The microgram amounts of DNA required as template are amplified from a single bacterium by a method called multiple displacement amplification (MDA) avoiding the need to grow cells. The ability to sequence DNA from individual cells will likely have an immense impact on microbiology considering the vast numbers of novel organisms, which have been inaccessible unless culture-independent methods could be used. However, special approaches have been necessary to work with amplified DNA. MDA may not recover the entire genome from the single copy present in most bacteria. Also, some sequence rearrangements can occur during the DNA amplification reaction. Over the past two years many research groups have begun to use MDA, and some practical approaches to single-cell sequencing have been developed. We review the consensus that is emerging on optimum methods, reliability of amplified template, and the proper interpretation of 'composite' genomes which result from the necessity of combining data from several single-cell MDA reactions in order to complete the assembly. Preferred laboratory methods are considered on the basis of experience at several large sequencing centers where >70% of genomes are now often recovered from single cells. Methods are reviewed for preparation of bacterial fractions from environmental samples, single-cell isolation, DNA amplification by MDA, and DNA sequencing.

  6. Investigation of Entamoeba histolytica in stool specimens by ELISA during a year

    Directory of Open Access Journals (Sweden)

    Mehmet Sinan Dal

    2011-03-01

    Full Text Available The aim of this study was to investigate Entamoebahistolytica in the stool samples that have beensent to microbiology laboratory by ELISA method.Materials and methods: This study was performed on800 stool specimens belonging to different age groupsand different patients sent to Microbiology Laboratory ofa State Hospital between January 2009 and December2009.Results: In this study Entamoeba histolytica/dispar cystsand/or trophozoites were observed in 31 (3.9% out of atotal of 800 stool specimens which were examined bynative-lugol. Entamoeba histolytica specific antigen wasinvestigated by ELISA in all stool specimens. Entamoebahistolytica specific antigen was detected in 12 (1.5% ofthese stool specimens. The diagnosis of amoebiasis forthe patients whose ELISA tests were “positive” and appropriatetherapy was begun.Conclusion: Entamoeba histolytica in the stool samplesshould be investigated and unnecessary treatmentsshould not given to patients. J Clin Exp Invest 2011; 2(1:50-54

  7. [First isolate of Encephalitozoon intestinalis from stools of a Colombian patient with AIDS].

    Science.gov (United States)

    Bedoya, Katherine; Montoya, Martha Nelly; Botero, Jorge; Galván, Ana Luz

    2008-09-01

    Microsporidia are obligate intracellular parasites that are recognized as important opportunistic pathogens of immunocompromised and transplanted patients. Enterocytozoon bieneusi and, less frequently, Encephalitozoon intestinalis are the most prevalent species in humans; both of them are associated with enteric infections. Cell cultures have been useful in the study of microsporidia biology. In Colombia, however, no isolates of microsporidia from patients with AIDS have been obtained. A cell culture of intestinal microsporidia was established from stools of positive patients in order to isolate a native strain. Stool from a single AIDS patient was concentrated with the water-ether technique, and the sediment was treated with a mixture of antibiotics and antifungal agents for 18 hours at 37 degrees C. Vero cells were cultivated in 24-well plates with Gibco RPMI medium supplemented with 10% bovine fetal serum and antibiotics. The culture was subsequently inoculated with previously concentrated spores. The medium was changed every second day and the presence of spores was evaluated with the Quick Hot Gram chromotrope stain. Two weeks post-infection, microsporidial spores were identified with characteristic morphology and staining properties. PCR results showed that Encephalitozoon intestinalis was the isolated species. A cell culture of microsporidia was established from a stool sample. This protocol is important to isolate and maintain additional native Colombian strains and it will contribute to biochemical, immunological and epidemiological studies of the currently established strain.

  8. High Prevalence of Campylobacter ureolyticus in Stool Specimens of Children with Diarrhea in Japan.

    Science.gov (United States)

    Hatanaka, Noritoshi; Shimizu, Akinori; Somroop, Srinuan; Li, Yiming; Asakura, Masahiro; Nagita, Akira; Prasad Awasthi, Sharda; Hinenoya, Atsushi; Yamasaki, Shinji

    2017-07-24

    Campylobacter ureolyticus has been considered as a potentially pathogenic bacterium. In this study, a total of 586 stool samples were collected from 0-12-year-old children with diarrhea between November 2013 and April 2015 and examined with microbiological tests in the hospital for the diagnosis of common enteric pathogens including C. jejuni and C. coli. Then in our laboratory, these samples were analyzed by 16S rRNA sequence-based Campylobacter genus-specific PCR (C16S PCR); 283 (48.3%) samples showed positive results with this PCR assay. Furthermore, C. ureolyticus was screened in these 283 samples by PCR assay, which can detect this species specifically. Surprisingly, C. ureolyticus was detected in 147 of the 283 C16S PCR-positive diarrheal stool samples (51.9%), which is much higher than the prevalence of C. jejuni and C. coli (15.5%), and 96 samples out of 147 were negative for any of the other enteric pathogens tested in the hospital; namely, C. ureolyticus was detected as a single pathogen in 96 samples. This finding suggests that C. ureolyticus may be a pathogen associated with diarrhea in children in Japan. To the best of our knowledge, this is the first report in which C. ureolyticus was detected among Japanese children with diarrhea.

  9. A Single Platinum Microelectrode for Identifying Soft Drink Samples

    Directory of Open Access Journals (Sweden)

    Lígia Bueno

    2012-01-01

    Full Text Available Cyclic voltammograms recorded with a single platinum microelectrode were used along with a non-supervised pattern recognition, namely, Principal Component Analysis, to conduct a qualitative analysis of sixteen different brands of carbonated soft drinks (Kuat, Soda Antarctica, H2OH!, Sprite 2.0, Guarana Antarctica, Guarana Antarctica Zero, Coca-Cola, Coca-Cola Zero, Coca-Cola Plus, Pepsi, Pepsi Light, Pepsi Twist, Pepsi Twist Light, Pepsi Twist 3, Schin Cola, and Classic Dillar’s. In this analysis, soft drink samples were not subjected to pre-treatment. Good differentiation among all the analysed soft drinks was achieved using the voltammetric data. An analysis of the loading plots shows that the potentials of −0.65 V, −0.4 V, 0.4 V, and 0.750 V facilitated the discrimination process. The electrochemical processes related to this potential are the reduction of hydrogen ions and inhibition of the platinum oxidation by the caffeine adsorption on the electrode surface. Additionally, the single platinum microelectrode was useful for the quality control of the soft drink samples, as it helped to identify the time at which the beverage was opened.

  10. Role of microscopic examination of stool specimens in the diagnosis ...

    African Journals Online (AJOL)

    stain of the stool in diagnosis of campylobacter infection, using culture as the gold standard. A total of 226 stool specimens were obtained from children with acute diarrhoea, attending Mulago Hospital in Kampala, Uganda. Stool smears were ...

  11. ‘Urmitella timonensis’ gen. nov., sp. nov., ‘Blautia marasmi’ sp. nov., ‘Lachnoclostridium pacaense’ sp. nov., ‘Bacillus marasmi’ sp. nov. and ‘Anaerotruncus rubiinfantis’ sp. nov., isolated from stool samples of undernourished African children

    Directory of Open Access Journals (Sweden)

    T.-P.-T. Pham

    2017-05-01

    Full Text Available We report here the main characteristics of five new species ‘Urmitella timonensis’ strain Marseille-P2918T (CSUR P2918, ‘Blautia marasmi’ strain Marseille-P2377T (CSUR P2377, ‘Lachnoclostridium pacaense’ strain Marseille-P3100T (CSUR P3100, ‘Bacillus marasmi’ strain Marseille-P3556T (CSUR P3556 and ‘Anaerotruncus rubiinfantis’ strain MT15T (CSUR P2276, which were isolated recently from stool samples taken from undernourished children in Niger and Senegal using microbial culturomics.

  12. Single-stage mass spectrometric analyses of resin bead samples

    Energy Technology Data Exchange (ETDEWEB)

    Smith, D. H.; Walker, R. L.; Bertram, L. K.; Carter, J. A.

    1978-10-01

    Plutonium and uranium from dissolver solutions loaded on resin beads can be analyzed on single-stage mass spectrometers with little or no degradation of results provided proper care is exercised with regard to sample handling techniques. Additionally, storage of samples on resin beads is feasible for periods at least as long as six months provided the beads are not exposed to residual HNO/sub 3/ and air; it is probable that beads will retain their integrity much longer than six months when stored under collodion, but as yet no data to support this contention have been collected. Conventional or commercial mass spectrometers can readily be adapted to the resin bead technique by installing a pulse-counting detection system. The cost of such conversion will vary depending on whether or not a data acquisition system will be needed. A reasonable estimate is that the cost will be in the neighborhood of $15,000; this figure includes the price of a multi-channel analyzer to serve as a temporary data storage device, but does not include the cost of a computer. It does not appear that it will be practicable to switch easily back and forth between pulse-counting and current integration modes unless the instrument is provided with a movable Faraday cup. Using the same multiplier in both modes would undoubtedly degrade its performance in each. The requirements of low background counting rates and high gain for pulse counting, and of relatively high signal handling capacity in current integration are mutually incompatible if demanded of the same multiplier.

  13. Bristol Stool Form Scale reliability and agreement decreases when determining Rome III stool form designations.

    Science.gov (United States)

    Chumpitazi, B P; Self, M M; Czyzewski, D I; Cejka, S; Swank, P R; Shulman, R J

    2016-03-01

    Rater reproducibility of the Bristol Stool Form Scale (BSFS), which categorizes stools into one of seven types, is unknown. We sought to determine reliability and agreement by individual stool type and when responses are categorized by Rome III clinical designation as normal or abnormal (constipation or diarrhea). Thirty-four gastroenterology providers from three institutions rated 35 stool photographs using the BSFS. Twenty rerated the photographs. 1190 individual stool type ratings were completed. Though only four photographs had absolute agreement (all Type 1 or Type 7), general agreement was high with 1132 (95.1%) of ratings being within one category type of the modal rating. Inter-rater and intra-rater reliability of the BSFS by individual stool type was excellent with intraclass correlations of 0.88 (95% CI: 0.86-0.90, p Bristol Stool Form Scale has excellent reliability and agreement when used to rate individual stool type by raters. However, BSFS reliability and agreement decreases when determining Rome III stool form categories. © 2015 John Wiley & Sons Ltd.

  14. Comparison of Escherichia coli O157:H7 antigen detection in stool and broth cultures to that in sorbitol-MacConkey agar stool cultures.

    Science.gov (United States)

    Stapp, J R; Jelacic, S; Yea, Y L; Klein, E J; Fischer, M; Clausen, C R; Qin, X; Swerdlow, D L; Tarr, P I

    2000-09-01

    We evaluated the Meridian IC-STAT direct fecal and broth culture antigen detection methods with samples from children infected with Escherichia coli O157:H7 and correlated the antigen detection results with the culture results. Stools of 16 children who had recently had stool cultures positive for this pathogen (population A) and 102 children with diarrhea of unknown cause (population B) were tested with the IC-STAT device (direct testing). Fecal broth cultures were also tested with this device (broth testing). The results were correlated to a standard of the combined yield from direct culture of stools on sorbitol-MacConkey (SMAC) agar and culture of broth on SMAC agar. Eleven (69%) of the population A stool specimens yielded E. coli O157:H7 when plated directly on SMAC agar. Two more specimens yielded this pathogen when the broth culture was similarly plated. Of these 13 stool specimens, 8 and 13 were positive by direct and broth testing (respective sensitivities, 62 and 100%). Compared to the sensitivity of a simultaneously performed SMAC agar culture, the sensitivity of direct testing was 73%. Three (3%) of the population B stool specimens contained E. coli O157:H7 on SMAC agar culture; one and three of these stool specimens were positive by direct and broth testing, respectively. The direct and broth IC-STAT tests were 100% specific with samples from children from population B. Direct IC-STAT testing of stools is rapid, easily performed, and specific but is insufficiently sensitive to exclude the possibility of infection with E. coli O157:H7. Performing the IC-STAT test with a broth culture increases its sensitivity. However, attempts to recover E. coli O157:H7 by culture should not be abandoned but, rather, should be increased when the IC-STAT test result is positive.

  15. Comparison of culture, single and multiplex real-time PCR for detection of Sabin poliovirus shedding in recently vaccinated Indian children.

    Science.gov (United States)

    Giri, Sidhartha; Rajan, Anand K; Kumar, Nirmal; Dhanapal, Pavithra; Venkatesan, Jayalakshmi; Iturriza-Gomara, Miren; Taniuchi, Mami; John, Jacob; Abraham, Asha Mary; Kang, Gagandeep

    2017-08-01

    Although, culture is considered the gold standard for poliovirus detection from stool samples, real-time PCR has emerged as a faster and more sensitive alternative. Detection of poliovirus from the stool of recently vaccinated children by culture, single and multiplex real-time PCR was compared. Of the 80 samples tested, 55 (68.75%) were positive by culture compared to 61 (76.25%) and 60 (75%) samples by the single and one step multiplex real-time PCR assays respectively. Real-time PCR (singleplex and multiplex) is more sensitive than culture for poliovirus detection in stool, although the difference was not statistically significant. © 2017 Wiley Periodicals, Inc.

  16. Amsterdam infant stool scale is more useful for assessing children who have not been toilet trained than Bristol stool scale

    NARCIS (Netherlands)

    Ghanma, A.; Puttemans, K.; Deneyer, M.; Benninga, M. A.; Vandenplas, Y.

    2014-01-01

    A validated stool scale would help both parents and clinicians to describe and differentiate between physiological and pathologic stool appearance. The Bristol Stool Scale (BSS), which was developed at the University of Bristol, is a medical aid that classifies human stools into seven categories

  17. Development of a Single-Sampling Noninvasive Hair Snare

    DEFF Research Database (Denmark)

    Bremner-Harrison, Samantha; Harrison, Stephen W. R.; Cypher, Brian L.

    2006-01-01

    Noninvasive hair and fecal DNA sampling provides a means of collecting information on elusive species, while causing little or no disturbance. However, current methods of hair collection do not preclude multiple sampling, thus risking sample contamination. We developed a hair snare that prevents ...

  18. Directional emission of single photons from small atomic samples

    DEFF Research Database (Denmark)

    Miroshnychenko, Yevhen; V. Poulsen, Uffe; Mølmer, Klaus

    2013-01-01

    We provide a formalism to describe deterministic emission of single photons with tailored spatial and temporal profiles from a regular array of multi-level atoms. We assume that a single collective excitation is initially shared by all the atoms in a metastable atomic state, and that this state i...... is coupled by a classical laser field to an optically excited state which rapidly decays to the ground atomic state. Our model accounts for the different field polarization components via re-absorption and emission of light by the Zeeman manifold of optically excited states.......We provide a formalism to describe deterministic emission of single photons with tailored spatial and temporal profiles from a regular array of multi-level atoms. We assume that a single collective excitation is initially shared by all the atoms in a metastable atomic state, and that this state...

  19. Development of a Single-Sampling Noninvasive Hair Snare

    DEFF Research Database (Denmark)

    Bremner-Harrison, Samantha; Harrison, Stephen W. R.; Cypher, Brian L.

    2006-01-01

    Noninvasive hair and fecal DNA sampling provides a means of collecting information on elusive species, while causing little or no disturbance. However, current methods of hair collection do not preclude multiple sampling, thus risking sample contamination. We developed a hair snare that prevents...... multiple sampling, is cost-effective, easy to construct, and safe for target and nontarget species. Our initial field tests on endangered San Joaquin kit foxes (Vulpes macrotis mutica) and swift foxes (Vulpes velox) suggest that this hair snare may be effective in collecting uncontaminated samples for DNA...

  20. Stool consistency is strongly associated with gut microbiota richness and composition, enterotypes and bacterial growth rates.

    Science.gov (United States)

    Vandeputte, Doris; Falony, Gwen; Vieira-Silva, Sara; Tito, Raul Y; Joossens, Marie; Raes, Jeroen

    2016-01-01

    The assessment of potentially confounding factors affecting colon microbiota composition is essential to the identification of robust microbiome based disease markers. Here, we investigate the link between gut microbiota variation and stool consistency using Bristol Stool Scale classification, which reflects faecal water content and activity, and is considered a proxy for intestinal colon transit time. Through 16S rDNA Illumina profiling of faecal samples of 53 healthy women, we evaluated associations between microbiome richness, Bacteroidetes:Firmicutes ratio, enterotypes, and genus abundance with self-reported, Bristol Stool Scale-based stool consistency. Each sample's microbiota growth potential was calculated to test whether transit time acts as a selective force on gut bacterial growth rates. Stool consistency strongly correlates with all known major microbiome markers. It is negatively correlated with species richness, positively associated to the Bacteroidetes:Firmicutes ratio, and linked to Akkermansia and Methanobrevibacter abundance. Enterotypes are distinctly distributed over the BSS-scores. Based on the correlations between microbiota growth potential and stool consistency scores within both enterotypes, we hypothesise that accelerated transit contributes to colon ecosystem differentiation. While shorter transit times can be linked to increased abundance of fast growing species in Ruminococcaceae-Bacteroides samples, hinting to a washout avoidance strategy of faster replication, this trend is absent in Prevotella-enterotyped individuals. Within this enterotype adherence to host tissue therefore appears to be a more likely bacterial strategy to cope with washout. The strength of the associations between stool consistency and species richness, enterotypes and community composition emphasises the crucial importance of stool consistency assessment in gut metagenome-wide association studies. Published by the BMJ Publishing Group Limited. For permission to

  1. Reliability and validity of a modified Bristol Stool Form Scale for children.

    Science.gov (United States)

    Lane, Mariella M; Czyzewski, Danita I; Chumpitazi, Bruno P; Shulman, Robert J

    2011-09-01

    This study sought to: evaluate the ability of children to reliably use a modified Bristol Stool Form Scale for Children (mBSFS-C), evaluate criterion-related validity of the mBSFS-C, and identify the lower age limit for mBSFS-C use. The mBSFS-C comprises 5 stool form types described and depicted in drawings. Children 3 to 18 years old rated stool form for 10 stool photographs. Because of low reliability when stool form descriptors were not read aloud (n = 119), a subsequent sample of children (n = 191) rated photographs with descriptors read. Intraclass correlation coefficients for descriptor-unread versus -read samples were 0.62 and 0.79, respectively. Children were increasingly reliable with age. Percentage of correct ratings varied with stool form type, but generally increased with age. With descriptors unread, children 8 years and older demonstrated acceptable interobserver reliability, with >78% of ratings correct. With descriptors read, children 6 years and older demonstrated acceptable reliability, with >80% of ratings correct. The mBSFS-C is reliable and valid for use by children, with age 6 years being the lower limit for scale use with descriptors read and age 8 years being the lower limit without descriptors read. We anticipate that the mBSFS-C can be effectively used in pediatric clinical and research settings. Copyright © 2011 Mosby, Inc. All rights reserved.

  2. Reliable single cell array CGH for clinical samples.

    Directory of Open Access Journals (Sweden)

    Zbigniew T Czyż

    Full Text Available BACKGROUND: Disseminated cancer cells (DCCs and circulating tumor cells (CTCs are extremely rare, but comprise the precursors cells of distant metastases or therapy resistant cells. The detailed molecular analysis of these cells may help to identify key events of cancer cell dissemination, metastatic colony formation and systemic therapy escape. METHODOLOGY/PRINCIPAL FINDINGS: Using the Ampli1™ whole genome amplification (WGA technology and high-resolution oligonucleotide aCGH microarrays we optimized conditions for the analysis of structural copy number changes. The protocol presented here enables reliable detection of numerical genomic alterations as small as 0.1 Mb in a single cell. Analysis of single cells from well-characterized cell lines and single normal cells confirmed the stringent quantitative nature of the amplification and hybridization protocol. Importantly, fixation and staining procedures used to detect DCCs showed no significant impact on the outcome of the analysis, proving the clinical usability of our method. In a proof-of-principle study we tracked the chromosomal changes of single DCCs over a full course of high-dose chemotherapy treatment by isolating and analyzing DCCs of an individual breast cancer patient at four different time points. CONCLUSIONS/SIGNIFICANCE: The protocol enables detailed genome analysis of DCCs and thereby assessment of the clonal evolution during the natural course of the disease and under selection pressures. The results from an exemplary patient provide evidence that DCCs surviving selective therapeutic conditions may be recruited from a pool of genomically less advanced cells, which display a stable subset of specific genomic alterations.

  3. Vapor and gas sampling of single-shell tank 241-U-104 using the in situ vapor sampling system

    International Nuclear Information System (INIS)

    Lockrem, L.L.

    1997-01-01

    The Vapor Issue.Resolution Program tasked the Vapor Team (VT) to collect representative headspace samples from Hanford Site single-shell tank (SST) 241-U-104. This document presents In Situ Vapor Sampling System (ISVS) data resulting from the July 16, 1996 sampling of SST 241-U-104. Analytical results will be presented in separate reports issued by the Pacific Northwest National Laboratory (PNNL) which supplied and analyzed the sample media

  4. Vapor and gas sampling of single-shell tank 241-BX-110 using the in situ vapor sampling system

    International Nuclear Information System (INIS)

    Lockrem, L.L.

    1997-01-01

    The Vapor Issue Resolution Program tasked the Vapor Team (the team) to collect representative headspace samples from Hanford Site single-shell tank (SST) 241-BX-110. This document presents sampling data resulting from the April 30, 1996 sampling of SST 241-BX-110. Analytical results will be presented in a separate report issued by Pacific Northwest National Laboratory (PNNL), which supplied and analyzed the sampling media

  5. Vapor and gas sampling of single-shell tank 241-S-106 using the in situ vapor sampling system

    International Nuclear Information System (INIS)

    Lockrem, L.L.

    1997-01-01

    The Vapor Issue Resolution Program tasked the Vapor Team (VT) to collect representative headspace samples from Hanford Site single-shell tank (SST) 241-S-106. This document presents In Situ vapor Sampling System (ISVS) data resulting from the June 13, 1996 sampling of SST 241-S-106. Analytical results will be presented in separate reports issued by the Pacific Northwest National Laboratory (PNNL) which'supplied and analyzed the sample media

  6. Vapor and gas sampling of single-shell tank 241-S-103 using the in situ vapor sampling system

    International Nuclear Information System (INIS)

    Lockrem, L.L.

    1997-01-01

    The Vapor Issue Resolution Program tasked the Vapor Team (VT) to collect representative headspace samples from Hanford Site single-shell tank (SST) 241-S-103. This document presents In Situ Vapor Sampling System (ISVS) data resulting from the June 12, 1996 sampling of SST 241-S-103. Analytical results will be presented in separate reports issued by the Pacific Northwest National Laboratory (PNNL) which supplied and analyzed the sample media

  7. Simple fecal flotation is a superior alternative to guadruple Kato Katz smear examination for the detection of hookworm eggs in human stool.

    Directory of Open Access Journals (Sweden)

    Tawin Inpankaew

    2014-12-01

    Full Text Available Microscopy-based identification of eggs in stool offers simple, reliable and economical options for assessing the prevalence and intensity of hookworm infections, and for monitoring the success of helminth control programs. This study was conducted to evaluate and compare the diagnostic parameters of the Kato-Katz (KK and simple sodium nitrate flotation technique (SNF in terms of detection and quantification of hookworm eggs, with PCR as an additional reference test in stool, collected as part of a baseline cross-sectional study in Cambodia.Fecal samples collected from 205 people in Dong village, Rovieng district, Preah Vihear province, Cambodia were subjected to KK, SNF and PCR for the detection (and in case of microscopy-based methods, quantification of hookworm eggs in stool. The prevalence of hookworm detected using a combination of three techniques (gold standard was 61.0%. PCR displayed a highest sensitivity for hookworm detection (92.0% followed by SNF (44.0% and quadruple KK smears (36.0% compared to the gold standard. The overall eggs per gram feces from SNF tended to be higher than for quadruple KK and the SNF proved superior for detecting low egg burdens.As a reference, PCR demonstrated the higher sensitivity compared to SNF and the quadruple KK method for detection of hookworm in human stool. For microscopic-based quantification, a single SNF proved superior to the quadruple KK for the detection of hookworm eggs in stool, in particular for low egg burdens. In addition, the SNF is cost-effective and easily accessible in resource poor countries.

  8. Simple fecal flotation is a superior alternative to guadruple Kato Katz smear examination for the detection of hookworm eggs in human stool.

    Science.gov (United States)

    Inpankaew, Tawin; Schär, Fabian; Khieu, Virak; Muth, Sinuon; Dalsgaard, Anders; Marti, Hanspeter; Traub, Rebecca J; Odermatt, Peter

    2014-12-01

    Microscopy-based identification of eggs in stool offers simple, reliable and economical options for assessing the prevalence and intensity of hookworm infections, and for monitoring the success of helminth control programs. This study was conducted to evaluate and compare the diagnostic parameters of the Kato-Katz (KK) and simple sodium nitrate flotation technique (SNF) in terms of detection and quantification of hookworm eggs, with PCR as an additional reference test in stool, collected as part of a baseline cross-sectional study in Cambodia. Fecal samples collected from 205 people in Dong village, Rovieng district, Preah Vihear province, Cambodia were subjected to KK, SNF and PCR for the detection (and in case of microscopy-based methods, quantification) of hookworm eggs in stool. The prevalence of hookworm detected using a combination of three techniques (gold standard) was 61.0%. PCR displayed a highest sensitivity for hookworm detection (92.0%) followed by SNF (44.0%) and quadruple KK smears (36.0%) compared to the gold standard. The overall eggs per gram feces from SNF tended to be higher than for quadruple KK and the SNF proved superior for detecting low egg burdens. As a reference, PCR demonstrated the higher sensitivity compared to SNF and the quadruple KK method for detection of hookworm in human stool. For microscopic-based quantification, a single SNF proved superior to the quadruple KK for the detection of hookworm eggs in stool, in particular for low egg burdens. In addition, the SNF is cost-effective and easily accessible in resource poor countries.

  9. Complexity of rice-water stool from patients with Vibrio cholerae plays a role in the transmission of infectious diarrhea.

    Science.gov (United States)

    Nelson, Eric J; Chowdhury, Ashrafuzzaman; Harris, Jason B; Begum, Yasmin A; Chowdhury, Fahima; Khan, Ashraful I; Larocque, Regina C; Bishop, Anne L; Ryan, Edward T; Camilli, Andrew; Qadri, Firdausi; Calderwood, Stephen B

    2007-11-27

    At the International Centre for Diarrhoeal Disease Research, Bangladesh, one-half of the rice-water stool samples that were culture-positive for Vibrio cholerae did not contain motile V. cholerae by standard darkfield microscopy and were defined as darkfield-negative (DF(-)). We evaluated the host and microbial factors associated with DF status, as well as the impact of DF status on transmission. Viable counts of V. cholerae in DF(-) stools were three logs lower than in DF(+) stools, although DF(-) and DF(+) stools had similar direct counts of V. cholerae by microscopy. In DF(-) samples, non-V. cholerae bacteria outnumbered V. cholerae 10:1. Lytic V. cholerae bacteriophage were present in 90% of DF(-) samples compared with 35% of DF(+) samples, suggesting that bacteriophage may limit culture-positive patients from producing DF(+) stools. V. cholerae in DF(-) and DF(+) samples were found both planktonically and in distinct nonplanktonic populations; the distribution of organisms between these compartments did not differ appreciably between DF(-) and DF(+) stools. This biology may impact transmission because epidemiological data suggested that household contacts of a DF(+) index case were at greater risk of infection with V. cholerae. We propose a model in which V. cholerae multiply in the small intestine to produce a fluid niche that is dominated by V. cholerae. If lytic phage are present, viable counts of V. cholerae drop, stools become DF(-), other microorganisms bloom, and cholera transmission is reduced.

  10. Human stool contains a previously unrecognized diversity of novel astroviruses

    Directory of Open Access Journals (Sweden)

    Zhao Guoyan

    2009-10-01

    Full Text Available Abstract Human astroviruses are a leading cause of gastrointestinal disease. Since their discovery in 1975, 8 closely related serotypes have been described in humans, and more recently, two new astrovirus species, astrovirus MLB1 and astrovirus VA1, were identified in diarrhea patients. In this study, we used consensus astrovirus primers targeting the RNA polymerase to define the diversity of astroviruses present in pediatric patients with diarrhea on two continents. From 416 stool specimens comprising two different cohorts from Vellore, India, 35 samples were positive. These positive samples were analyzed further by either sequencing of the ~400 bp amplicon generated by the consensus PCR or by performing additional RT-PCR specific for individual astroviruses. 19 samples contained the classic human astrovirus serotypes 1-8 while 7 samples were positive for the recently described astrovirus MLB1. Strikingly, from samples that were positive in the consensus PCR screen but negative in the specific PCR assays, five samples contained sequences that were highly divergent from all previously described astroviruses. Sequence analysis suggested that three novel astroviruses, tentatively named astroviruses VA2, MLB2 and VA3, were present in these five patient specimens (AstV-VA2 in 2 patients, AstV-MLB2 in 2 patients and AstV-VA3 in one patient. Using the same RT-PCR screening strategy, 13 samples out of 466 tested stool specimens collected in St. Louis, USA were positive. Nine samples were positive for the classic human astroviruses. One sample was positive for AstV-VA2, and 3 samples were positive for AstV-MLB2 demonstrating that these two viruses are globally widespread. Collectively, these findings underscore the tremendous diversity of astroviruses present in fecal specimens from diarrhea patients. Given that a significant fraction of diarrhea etiologies is currently unknown, it is plausible that these or other yet unrecognized astroviruses may be

  11. Effects of olestra and sorbitol consumption on objective measures of diarrhea: impact of stool viscosity on common gastrointestinal symptoms.

    Science.gov (United States)

    McRorie, J; Zorich, N; Riccardi, K; Bishop, L; Filloon, T; Wason, S; Giannella, R

    2000-02-01

    The aim of this study was to determine the effects of olestra and sorbitol consumption on three accepted objective measures of diarrhea (stool output >250 g/day, liquid/watery stools, bowel movement frequency >3/day), and how stool composition influences reports of common gastrointestinal symptoms. A double-blind, placebo-controlled study compared the effects of sorbitol (40 g/day in candy), a poorly absorbed sugar-alcohol with known osmotic effects, with those of olestra (20 or 40 g/day in potato chips), a nonabsorbed fat, on objective measures of stool composition and GI symptoms. Sixty-six subjects resided on a metabolic ward for 12 days: 2 days lead-in, 4 days baseline, 6 days treatment. Sorbitol 40 g/day resulted in loose/liquid stools within 1-3 h of consumption. In contrast, olestra resulted in a dose-responsive stool softening effect after 2-4 days of consumption. Subjects reported "diarrhea" when mean stool apparent viscosity (peak force (PF), g) decreased from a perceived "normal" (mean +/- SE, 1355 +/- 224 g PF; firm stool) to loose (260 +/- 68 g PF) stool. Mean apparent viscosity of stool during treatment: placebo, 1363 +/- 280 g (firm); olestra 20 g/day 743 +/- 65 g (soft); olestra 40 g/day, 563 +/- 105 g (soft); and sorbitol 40 g/day, 249 +/- 53 g (loose). Of the 1098 stool samples collected, 38% (419/1098) were rated by subjects as "diarrhea," yet only 2% of treatment days (all in the sorbitol treatment group) met commonly accepted criteria for a clinical diarrhea. Sorbitol, but not olestra, increased the severity of abdominal cramping, urgency and nausea compared to placebo. Olestra consumption, at levels far in excess of normal snacking conditions, resulted in a gradual stool softening effect after several days of consumption, did not meet any of the three objective measures of diarrhea, and did not increase GI symptoms. Sorbitol consumption, at only 80% of the dose requiring a "laxative effect" information label, resulted in rapid onset loose

  12. Molecular Detection of Strongyloides ratti in Faecal Samples from ...

    African Journals Online (AJOL)

    for all collected stool samples. Briefly, 2 g of each sample was placed in the center of plastic petri dish containing approximately 20 mL of nutrient agar. Dishes were sealed with parafilm tape to prevent larvae from crawling .... loops spiraling around the intestine or running parallel with intestine. Eggs in single row in uterus.

  13. Influence of selected stool concentration techniques on the effectiveness of PCR examination in Giardia intestinalis diagnostics.

    Science.gov (United States)

    Stojecki, K; Sroka, J; Karamon, J; Kusyk, P; Cencek, T

    2014-01-01

    Giardia intestinalis is a widespread parasitic protozoa which has great significance as a public health threat. Molecular diagnostics of stool sample can be unreliable because of the presence of inhibitors of enzymatic reactions. The aim of this study was to determine the effectiveness of selected pre-treatment methods of fecal samples for further PCR-based diagnostics of G. intestinalis, and the effect of each component of pre-treatment solutions on PCR reactions. Seven stool concentration techniques were compared. The results showed that the most efficient concentration method for stool sample preparation for detection of G. intestinalis by PCR is centrifugal flotation with Percoll (with saturated NaNO3 as the flotation solution). This method is relatively inexpensive, less labor-intensive, and suitable for epidemiological monitoring and clinical investigations.

  14. Effects of sample treatments on genome recovery via single-cell genomics

    Energy Technology Data Exchange (ETDEWEB)

    Clingenpeel, Scott [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Schwientek, Patrick [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Hugenholtz, Philip [Univ. of Queensland, Brisbane (Australia); Woyke, Tanja [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)

    2014-06-13

    It is known that single-cell genomics is a powerful tool for accessing genetic information from uncultivated microorganisms. Methods of handling samples before single-cell genomic amplification may affect the quality of the genomes obtained. Using three bacterial strains we demonstrate that, compared to cryopreservation, lower-quality single-cell genomes are recovered when the sample is preserved in ethanol or if the sample undergoes fluorescence in situ hybridization, while sample preservation in paraformaldehyde renders it completely unsuitable for sequencing.

  15. Brazilian Portuguese translation, cross-cultural adaptation and reproducibility assessment of the modified Bristol Stool Form Scale for children.

    Science.gov (United States)

    Jozala, Debora Rodrigues; Oliveira, Isabelle Stefan de Faria; Ortolan, Erika Veruska Paiva; Oliveira Junior, Wilson Elias de; Comes, Giovana Tuccille; Cassettari, Vanessa Mello Granado; Self, Mariella Marie; Lourenção, Pedro Luiz Toledo de Arruda

    2018-03-15

    To translate and culturally adapt the modified Bristol Stool Form Scale for children into Brazilian Portuguese, and to evaluate the reproducibility of the translated version. The stage of translation and cross-cultural adaptation was performed according to an internationally accepted methodology, including the translation, back-translation, and pretest application of the translated version to a sample of 74 children to evaluate the degree of understanding. The reproducibility of the translated scale was assessed by applying the final version of Brazilian Portuguese modified Bristol Stool Form Scale for children to a sample of 64 children and 25 healthcare professionals, who were asked to correlate a randomly selected description from the translated scale with the corresponding representative illustration of the stool type. The final version of Brazilian Portuguese modified Bristol Stool Form Scale for children were evidently reproducible, since almost complete agreement (k>0,8) was obtained among the translated descriptions and illustrations of the stool types, both among the children and the group of specialists. The Brazilian Portuguese modified Bristol Stool Form Scale for children was shown to be reliable in providing very similar results for the same respondents at different times and for different examiners. The Brazilian Portuguese modified Bristol Stool Form Scale for children is reproducible; it can be applied in clinical practice and in scientific research in Brazil. Copyright © 2018 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  16. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens

    NARCIS (Netherlands)

    Schuurman, T.; Lankamp, P.; van Belkum, A.; Kooistra-Smid, M.; van Zwet, A.

    2007-01-01

    Giardia lamblia is one of the most common intestinal parasites worldwide, with microscopy being the diagnostic reference standard for use with human stools. However, microscopy is time-consuming, labour-intensive and lacks sensitivity when single stools are examined. In the present study,

  17. Validity and reliability of the Bristol Stool Form Scale in healthy adults and patients with diarrhoea-predominant irritable bowel syndrome.

    Science.gov (United States)

    Blake, M R; Raker, J M; Whelan, K

    2016-10-01

    The Bristol Stool Form Scale (BSFS) is a 7-point scale used extensively in clinical practice and research for stool form measurement, which has undergone limited validity and reliability testing. To determine the validity and reliability of the BSFS in measuring stool form in healthy adults and patients with diarrhoea-predominant irritable bowel syndrome (IBS-D). One hundred and sixty-nine healthy volunteers provided a stool sample and used the BSFS to classify stool form, which was compared with measured stool water content and with values from 19 patients with IBS-D. Eighty-six volunteers used the BSFS to classify 26 stool models to determine accuracy and reliability. Volunteers' classifications of stool type correlated with stool water (Spearman's rho = 0.491, P < 0.001), which increased in hard (Types 1-2), normal (Types 3-5) and loose stools (Types 6-7) (P < 0.001). The BSFS detected differences in stool form between healthy volunteers (mean 3.7, s.d. 1.5) and IBS-D patients (mean 5.0, s.d. 1.2) (P < 0.001). Overall, 977/1204 (81%) stool models were correctly classified (substantial accuracy, κ = 0.78), although <80% of Types 2, 3, 5 and 6 were classified correctly. On 852/1118 (76%) occasions, volunteers classified covert duplicate models to the same stool type (substantial reliability, κ = 0.72), but with only moderate reliability for Types 2 (63%, κ = 0.57) and 3 (62%, κ = 0.55). The BSFS demonstrated substantial validity and reliability, although difficulties arose around clinical decision points (Types 2, 3, 5, 6) that warrant investigation in larger clinical populations. Potential for improving validity and reliability through modifications to the BSFS or training in its use should be explored. © 2016 John Wiley & Sons Ltd.

  18. Comparison of the compositions of the stool microbiotas of infants fed goat milk formula, cow milk-based formula, or breast milk.

    Science.gov (United States)

    Tannock, Gerald W; Lawley, Blair; Munro, Karen; Gowri Pathmanathan, Siva; Zhou, Shao J; Makrides, Maria; Gibson, Robert A; Sullivan, Thomas; Prosser, Colin G; Lowry, Dianne; Hodgkinson, Alison J

    2013-05-01

    The aim of the study was to compare the compositions of the fecal microbiotas of infants fed goat milk formula to those of infants fed cow milk formula or breast milk as the gold standard. Pyrosequencing of 16S rRNA gene sequences was used in the analysis of the microbiotas in stool samples collected from 90 Australian babies (30 in each group) at 2 months of age. Beta-diversity analysis of total microbiota sequences and Lachnospiraceae sequences revealed that they were more similar in breast milk/goat milk comparisons than in breast milk/cow milk comparisons. The Lachnospiraceae were mostly restricted to a single species (Ruminococcus gnavus) in breast milk-fed and goat milk-fed babies compared to a more diverse collection in cow milk-fed babies. Bifidobacteriaceae were abundant in the microbiotas of infants in all three groups. Bifidobacterium longum, Bifidobacterium breve, and Bifidobacterium bifidum were the most commonly detected bifidobacterial species. A semiquantitative PCR method was devised to differentiate between B. longum subsp. longum and B. longum subsp. infantis and was used to test stool samples. B. longum subsp. infantis was seldom present in stools, even of breast milk-fed babies. The presence of B. bifidum in the stools of breast milk-fed infants at abundances greater than 10% of the total microbiota was associated with the highest total abundances of Bifidobacteriaceae. When Bifidobacteriaceae abundance was low, Lachnospiraceae abundances were greater. New information about the composition of the fecal microbiota when goat milk formula is used in infant nutrition was thus obtained.

  19. Adaptation and validation of the Bristol scale stool form translated into the Spanish language among health professionals and patients.

    Science.gov (United States)

    Parés, D; Comas, M; Dorcaratto, D; Araujo, M I; Vial, M; Bohle, B; Pera, M; Grande, L

    2009-05-01

    stool type represents an important semiologic part of medical interviews. The Bristol Scale Stool Form is a clinical tool to evaluate stool consistency and form. The aim of this study was to translate and adapt the Bristol Scale Stool Form into Spanish. Differences in validation results between health professionals and patients surveyed were also evaluated. the study population included 79 physicians, 79 nurses, and 78 patients. Subjects were invited to match a randomly selected text defining one of the seven stool types in the scale with one of seven drawings described originally. A random selection of samples was offered for re-test reliability. the overall Kappa index was 0.708. Thirty-two subjects repeated the test for a test-retest assessment in a mean interval of 7.76 days, and the percentage concordance between definition and image was 84.4% with a Kappa index of 0.816. There were no differences in the validation study between physicians, nurses, and patients. this study has shown that the Spanish version of the Bristol Scale Stool Form is reliable for use as a tool to evaluate stool consistency and form.

  20. Standard Specification for Sampling Single-Phase Geothermal Liquid or Steam for Purposes of Chemical Analysis

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    1983-01-01

    1.1 This specification covers the basic requirements for equipment to be used for the collection of uncontaminated and representative samples from single-phase geothermal liquid or steam. Geopressured liquids are included. See Fig 1.

  1. Optimal sampling strategies to assess inulin clearance in children by the inulin single-injection method

    NARCIS (Netherlands)

    van Rossum, Lyonne K.; Mathot, Ron A. A.; Cransberg, Karlien; Vulto, Arnold G.

    2003-01-01

    Glomerular filtration rate in patients can be determined by estimating the plasma clearance of inulin with the single-injection method. In this method, a single bolus injection of inulin is administered and several blood samples are collected. For practical and convenient application of this method

  2. A Novel Astrovirus-Like RNA Virus Detected in Human Stool

    NARCIS (Netherlands)

    Oude Munnink, Bas B.; Cotten, Matthew; Canuti, Marta; Deijs, Martin; Jebbink, Maarten F.; van Hemert, Formijn J.; Phan, My V. T.; Bakker, Margreet; Jazaeri Farsani, Seyed Mohammad; Kellam, Paul; van der Hoek, Lia

    2016-01-01

    Several novel clades of astroviruses have recently been identified in human faecal samples. Here, we describe a novel astrovirus-like RNA virus detected in human stools, which we have tentatively named bastrovirus. The genome of this novel virus consists of 6,300 nucleotides organized in three open

  3. Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method

    NARCIS (Netherlands)

    van Gool, T.; Canning, E. U.; Gilis, H.; van den Bergh Weerman, M. A.; Eeftinck Schattenkerk, J. K.; Dankert, J.

    1994-01-01

    Two species of microsporidia, Enterocytozoon bieneusi and Septata intestinalis have been reported as intestinal parasites of AIDS patients. In attempts to establish E. bieneusi in vitro, spores were concentrated from stool samples from 4 AIDS patients with biopsy-proven E. bieneusi infections. After

  4. Quality control of parasitology stool examination in Tabriz clinical laboratories

    Directory of Open Access Journals (Sweden)

    shahram Khademvatan

    2011-06-01

    Full Text Available The purpose of quality control program was to make doctors and laboratory personnel trust in laboratory results and consequently increasing confidence in laboratory achievements. The quality assurance means raising the level of quality in all tests that lead to raising the level of work efficiency and laboratories including minimum expense for society and minimum time for lab personnel. This study aimed to assess and determine the accuracy and precision of results in Tabriz medical diagnostic laboratories. Materials and Methods: In this retrospective study, 790 stool samples were selected randomly and tested by standard methods.Student t- test, SPSS software and sensitivity and accuracy formulas were used for data analysis. Results: The sensitivity was 62%, 22% and 8% with 95% confidence intervals for worm's eggs, protozoan cysts and trophozoite detection respectively. Conclusion: To elevate quality assurance in clinical diagnostic laboratory, monitoring and check of the laboratories by standard methods continually should be done.

  5. Digital quantification of gene methylation in stool DNA by emulsion-PCR coupled with hydrogel immobilized bead-array.

    Science.gov (United States)

    Liu, Yunlong; Wu, Haiping; Zhou, Qiang; Song, Qinxin; Rui, Jianzhong; Zou, Bingjie; Zhou, Guohua

    2017-06-15

    Aberrations of gene methylation in stool DNA (sDNA) is an effective biomarker for non-invasive colorectal cancer diagnosis. However, it is challenging to accurately quantitate the gene methylation levels in sDNA due to the low abundance and degradation of sDNA. In this study, a digital quantification strategy was proposed by combining emulsion PCR (emPCR) with hydrogel immobilized bead-array. The assay includes following steps: bisulfite conversion of sDNA, pre-amplification by PCR with specific primers containing 5' universal sequences, emPCR of pre-amplicons with beaded primers to achieve single-molecular amplification and identification of hydrogel embedding beads coated with amplicons. The sensitivity and the specificity of the method are high enough to pick up 0.05% methylated targets from unmethylated DNA background. The successful detection of hypermethylated vimentin gene in clinical stool samples suggests that the proposed method should be a potential tool for non-invasive colorectal cancer screening. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Noninvasive detection through REMS-PCR technique of K-ras mutations in stool DNA of patients with colorectal cancer.

    Science.gov (United States)

    Mixich, Francisc; Ioana, Mihai; Voinea, Florea; Săftoiu, Adrian; Ciurea, Tudorel

    2007-03-01

    Tumor exfoliated cells that shed into stool are attractive targets for molecular screening and early detection of colon malignancies. Many studies have suggested that the detection of activated ras may have diagnostic or prognostic importance. The aim of this study was to establish the suitability for use in diagnostic laboratories of the noninvasive screening test of K-ras mutation determination in the stool and its routine prognostic value in colorectal cancer. Paired stool and tissue specimens obtained after polypectomy and colorectal biopsy from 28 patients diagnosed solely by histopathological findings with primary colorectal carcinoma, were prospectively studied for K-ras codon 12 mutations by restriction endonuclease-mediated selective (REMS)-PCR. DNA was obtained in 28 of tissue samples (100%) and 26 of stool samples (92.8%). K-ras codon 12 mutation was seen in 14 (50.0%) paired stool and tissue samples. Mutation detection was possible in 1000-fold excess of wild-type sequence. These results may be important in the design of genetic screening programs, determination of prognosis, early detection and treatment for patients with colon malignancy. The sensitivity and specificity of K-ras determination on stool-derived DNA in patients with colorectal carcinoma, support the opportunity of a large-scale trial to validate its use as a screening test. REMS- PCR is not labor intensive, but a sensitive, rapid, and robust assay for the detection of point mutations, and was introduced by us in a routine diagnostic laboratory.

  7. Prevalence of Salmonella Excretion in Stool: A Community Survey in 2 Sites, Guinea-Bissau and Senegal.

    Science.gov (United States)

    Im, Justin; Nichols, Chelsea; Bjerregaard-Andersen, Morten; Sow, Amy Gassama; Løfberg, Sandra; Tall, Adama; Pak, Gi Deok; Aaby, Peter; Baker, Stephen; Clemens, John D; Espinoza, Ligia Maria Cruz; Konings, Frank; May, Jürgen; Monteiro, Mario; Niang, Aissatou; Panzner, Ursula; Park, Se Eun; Schütt-Gerowitt, Heidi; Wierzba, Thomas F; Marks, Florian; von Kalckreuth, Vera

    2016-03-15

    Chronic and convalescent carriers play an important role in the transmission and endemicity of many communicable diseases. A high incidence of Salmonella enterica serovar Typhi and invasive nontyphoidal Salmonella (NTS) infection has been reported in parts of sub-Saharan Africa, yet the prevalence of Salmonella excretion in the general population is unknown. Stool specimens were collected from a random sample of households in 2 populations in West Africa: Bissau, Guinea-Bissau, and Dakar, Senegal. Stool was cultured to detect presence of Salmonella, and antimicrobial susceptibility testing was performed on the isolated organisms. Stool was cultured from 1077 and 1359 individuals from Guinea-Bissau and Senegal, respectively. Salmonella Typhi was not isolated from stool samples at either site. Prevalence of NTS in stool samples was 24.1 (95% confidence interval [CI], 16.5-35.1; n = 26/1077) per 1000 population in Guinea-Bissau and 10.3 (95% CI, 6.1-17.2; n = 14/1359) per 1000 population in Senegal. Evidence of NTS excretion in stool in both study populations indicates a possible NTS transmission route in these settings. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  8. Analysis of U and Pu resin bead samples with a single stage mass spectrometer

    International Nuclear Information System (INIS)

    Smith, D.H.; Walker, R.L.; Bertram, L.K.; Carter, J.A.

    1979-01-01

    Resin bead sampling enables the shipment of nanogram U and Pu quantities for analysis. Application of this sampling technique to safeguards was investigated with a single-stage mass spectrometer. Standards gave results in good agreement with NBS certified values. External precisions of +-0.5% were obtained on isotopic ratios of approx. 0.01; precisions on quantitative measurements are +-1.0%

  9. Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples

    NARCIS (Netherlands)

    Catsburg, Arnold; van der Zwet, Wil C.; Morre, Servaas A.; Ouburg, Sander; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

    2007-01-01

    Reliable analysis of single nucleotide polymorphisms (SNPs) in DNA derived from samples containing low numbers of cells or from suboptimal sources can be difficult. A new procedure to characterize multiple SNPs in traces of DNA from plasma and old dried blood samples was developed. Six SNPs in the

  10. Quantitative Single-letter Sequencing: a method for simultaneously monitoring numerous known allelic variants in single DNA samples

    Directory of Open Access Journals (Sweden)

    Duborjal Hervé

    2008-02-01

    Full Text Available Abstract Background Pathogens such as fungi, bacteria and especially viruses, are highly variable even within an individual host, intensifying the difficulty of distinguishing and accurately quantifying numerous allelic variants co-existing in a single nucleic acid sample. The majority of currently available techniques are based on real-time PCR or primer extension and often require multiplexing adjustments that impose a practical limitation of the number of alleles that can be monitored simultaneously at a single locus. Results Here, we describe a novel method that allows the simultaneous quantification of numerous allelic variants in a single reaction tube and without multiplexing. Quantitative Single-letter Sequencing (QSS begins with a single PCR amplification step using a pair of primers flanking the polymorphic region of interest. Next, PCR products are submitted to single-letter sequencing with a fluorescently-labelled primer located upstream of the polymorphic region. The resulting monochromatic electropherogram shows numerous specific diagnostic peaks, attributable to specific variants, signifying their presence/absence in the DNA sample. Moreover, peak fluorescence can be quantified and used to estimate the frequency of the corresponding variant in the DNA population. Using engineered allelic markers in the genome of Cauliflower mosaic virus, we reliably monitored six different viral genotypes in DNA extracted from infected plants. Evaluation of the intrinsic variance of this method, as applied to both artificial plasmid DNA mixes and viral genome populations, demonstrates that QSS is a robust and reliable method of detection and quantification for variants with a relative frequency of between 0.05 and 1. Conclusion This simple method is easily transferable to many other biological systems and questions, including those involving high throughput analysis, and can be performed in any laboratory since it does not require specialized

  11. PoopMD, a Mobile Health Application, Accurately Identifies Infant Acholic Stools.

    Directory of Open Access Journals (Sweden)

    Amy Franciscovich

    Full Text Available Biliary atresia (BA is the leading cause of pediatric end-stage liver disease in the United States. Education of parents in the perinatal period with stool cards depicting acholic and normal stools has been associated with improved time-to-diagnosis and survival in BA. PoopMD is a mobile application that utilizes a smartphone's camera and color recognition software to analyze an infant's stool and determine if additional follow-up is indicated. PoopMD was developed using custom HTML5/CSS3 and wrapped to work on iOS and Android platforms. In order to define the gold standard regarding stool color, seven pediatricians were asked to review 45 photographs of infant stool and rate them as acholic, normal, or indeterminate. Samples for which 6+ pediatricians demonstrated agreement defined the gold standard, and only these samples were included in the analysis. Accuracy of PoopMD was assessed using an iPhone 5s with incandescent lighting. Variability in analysis of stool photographs as acholic versus normal with intermediate rating weighted as 50% agreement (kappa was compared between three laypeople and one expert user. Variability in output was also assessed between an iPhone 5s and a Samsung Galaxy S4, as well as between incandescent lighting and compact fluorescent lighting. Six-plus pediatricians agreed on 27 normal and 7 acholic photographs; no photographs were defined as indeterminate. The sensitivity was 7/7 (100%. The specificity was 24/27 (89% with 3/27 labeled as indeterminate; no photos of normal stool were labeled as acholic. The Laplace-smoothed positive likelihood ratio was 6.44 (95% CI 2.52 to 16.48 and the negative likelihood ratio was 0.13 (95% CI 0.02 to 0.83. kappauser was 0.68, kappaphone was 0.88, and kappalight was 0.81. Therefore, in this pilot study, PoopMD accurately differentiates acholic from normal color with substantial agreement across users, and almost perfect agreement across two popular smartphones and ambient light

  12. Helicobacter Pylori Stool Antigen Assay in Hyperemesis Gravidarum.

    Science.gov (United States)

    Kabir, S; Khanam, R A; Basher, M S; Azam, M S; Hossain, M A; Mirza, T T; Banu, K A; Karmoker, R K

    2017-04-01

    Hyperemesis gravidarum is the most severe form of nausea and vomiting in pregnancy that seriously affects the pregnancy outcome. It is a disease with unknown etiology and varieties of contributing factors like hormonal changes, psychological and immunological factors. A significantly high prevalence of Helicobacter pylori among pregnant women with Hyperemesis gravidarum has been revealed recently. A descriptive, cross-sectional study was carried out at antenatal ward, Department of Obstetrics and Gynaecology, Mymensingh Medical College Hospital, Mymensingh for a period of twenty-one months among purposively selected thirty-six patients with Hyperemesis gravidarum with a view to assess the involvement of H. pylori in Hyperemesis gravidarum. Data were collected through interview, physical examinations and laboratory investigations by using case record form. Statistical analysis was performed using SPSS version 20.0 for Windows. Highest number 16(44.44%) of respondents were in age group 20 to 24 years with a mean of 23.81 years and a standard deviation (SD) of 4.55 years. Majority 29(80.56%) of the women had education less than 12 years, as many as 28(77.78%) women were housewives, and at least 14(38.89%) women had unplanned pregnancies. An overwhelming majority 29(80.56%) of women had their pregnancy duration between 8 to 12 weeks with a mean duration of 10.64 weeks and a standard deviation of 2.35 weeks. Majority 20(55.56%) of women were pregnant for first time, as many as 19(52.78%) women had duration of illness for 5 to 9 weeks. Of 16 multi-gravid women, 7(43.75%) had history of similar condition in their previous pregnancies. As many as 9 (25.00%) women had family history of similar condition in their mothers and sisters. First trimester was time of manifestation of the condition.At least 11 (30.56%) stool samples were positive for H. pylori stool antigen. Family history of Hyperemesis gravidarum and presence of H. pylori stool antigen are statistically

  13. Isolation and characterization of Yersinia-specific bacteriophages from pig stools in Finland.

    Science.gov (United States)

    Salem, M; Virtanen, S; Korkeala, H; Skurnik, M

    2015-03-01

    Bacteriophages infect bacteria, and they are present everywhere in the world including the intestinal tracts of animals. Yersiniosis is a common foodborne infection caused by Yersinia enterocolitica and Yersinia pseudotuberculosis. As these bacteria are frequently isolated from pigs, we wanted to know whether Yersinia-specific bacteriophages are also present in the pig stools and, if so, whether there is a positive or negative association between the prevalence of the Yersinia phages and the pathogenic Yersinia in the stool samples. Altogether 793 pig stool samples collected between November 2010 and March 2012 from 14 Finnish pig farms were screened for the presence of bacteriophages able to infect Y. enterocolitica serotype O:3, O:5,27 or O:9 strains, or Y. pseudotuberculosis serotype O:1a, O:1b or O:3 strains. Yersinia phages were isolated from 90 samples from eight farms. Yersinia enterocolitica O:3 was infected by 59 phages, 28 phages infected serotypes O:3 and O:5,27, and eight phages infected serotypes O:3, O:5,27 and O:9, and Y. pseudotuberculosis O:1a by eight phages. Many phages originating from pigs in the same farm were identical based on their restriction enzyme digestion patterns; 20 clearly different phages were selected for further characterization. Host ranges of these phages were tested with 94 Yersinia strains. Six of the phages infected eight strains, 13 phages infected three strains, and one phage infected only one strain, indicating that the phages had a relatively narrow host range. There was a clear association between the presence of the host bacteria and specific phages in the stools. The isolated bacteriophages may have potential as biocontrol agents for yersiniosis in both humans and pigs in future, and as alternatives or in addition to antibiotics. To our knowledge, this is the first reported isolation of Yersinia-specific phages from pig stool samples. © 2014 The Society for Applied Microbiology.

  14. Identification and genetic characterization of a novel circular single-stranded DNA virus in a human upper respiratory tract sample.

    Science.gov (United States)

    Cui, Lunbiao; Wu, Binyao; Zhu, Xiaojuan; Guo, Xiling; Ge, Yiyue; Zhao, Kangchen; Qi, Xian; Shi, Zhiyang; Zhu, Fengcai; Sun, Lixin; Zhou, Minghao

    2017-11-01

    Metagenomic analysis through high-throughput sequencing is a tool for detecting both known and novel viruses. Using this technique, a novel circular single-stranded DNA (ssDNA) virus genome was discovered in respiratory secretions from a febrile traveler. The virus, named human respiratory-associated PSCV-5-like virus (HRAPLV), has a genome comprising 3,018 bases, with two major putative ORFs inversely encoding capsid (Cap) and replicase (Rep) protein and separated by two intergenic regions. One stem-loop structure was predicted in the larger intergenic region (LIR). The predicted amino acid sequences of the Cap and Rep proteins of HRAPLV showed highest identity to those of porcine stool-associated circular virus 5 isolate CP3 (PoSCV 5) (53.0% and 48.9%, respectively). The host tropism of the virus is unknown, and further study is warranted to determine whether this novel virus is associated with human disease.

  15. Symptomatic Diverticulosis Is Characterized By Loose Stools.

    Science.gov (United States)

    Järbrink-Sehgal, M Ellionore; Andreasson, Anna; Talley, Nicholas J; Agréus, Lars; Song, Jeong-Yeop; Schmidt, Peter T

    2016-12-01

    Symptomatic uncomplicated diverticular disease is considered to be a discreet clinical entity distinct from irritable bowel syndrome (IBS), but population-based data are unavailable. We aimed to investigate the prevalence and location of diverticulosis in the general population, and its association with colonic symptoms and mental health. We propose that individuals with diverticulosis would report more constipation and IBS. We performed a population-based study of randomly selected adults born in Sweden (age, 18-70 y; 57.2% women); 745 received a gastroenterology consultation, completed validated abdominal symptom and mental health questionnaires, and were examined by colonoscopy. Logistic regression was used to calculate the associations between diverticulosis and age, sex, gastrointestinal symptoms, anxiety, depression, and self-rated health. Among the 742 participants (54.6% women), 130 (17.5%) had diverticulosis. Age was the strongest predictor of diverticulosis (P diverticulosis was rare in participants younger than 40 years (0.7%). All participants with diverticulosis had sigmoid involvement. Participants with diverticulosis were more likely to report loose stools (odds ratio [OR], 1.88; 95% confidence interval [CI], 1.20-2.96), urgency (OR, 1.64; 95% CI, 1.02-2.63), passing mucus (OR, 2.26; 95% CI, 1.08-4.72), and a high stool frequency (OR, 2.02; 95% CI, 1.11-3.65). Diverticulosis was associated with abdominal pain (OR, 2.10; 95% CI, 1.01-4.36; P = .047) and diarrhea-predominant IBS (OR, 9.55; 95% CI, 1.08-84.08; P = .04) in participants older than 60 years. The presence of anxiety and depression and self-rated health were similar in participants with and without diverticulosis. The prevalence of diverticulosis is age-dependent. Diverticulosis is associated with diarrhea in subjects across all age ranges. In subjects older than age 60, diverticulosis is associated with abdominal pain and diarrhea-predominant IBS. Copyright © 2016 AGA Institute

  16. Detection of Helicobacter pylori by Real-Time PCR for 16s rRNA in Stools of NonInfected Healthy Children, Using ELISA Antigen Stool Test as the Gold Standard.

    Science.gov (United States)

    George, Sergio; Mamani, Nora; Lucero, Yalda; Torres, Juan Pablo; Farfán, Mauricio; Lagomarcino, Anne J; Orellana, Andrea; O'Ryan, Miguel

    2016-12-01

    We previously detected Helicobacter pylori infection by stool antigen ELISA assay in 33-41% of asymptomatic Chilean children between 2-3 years of age, of which 11-20% had a transient infection and 21-22% a persistent infection. A total of 88% of ELISA-positive samples were also rtPCR positive, while 37/133 (33%) of ELISA-negative stool samples were rtPCR positive. The significance of a ELISA-negative/rtPCR-positive sample requires clarification. We aimed to determine whether rtPCR is able to detect persistent infections not detected by ELISA. We selected 36 children with an ELISA-negative/rtPCR-positive stool sample, of which 25 were never H. pylori infected according to ELISA, and 11 had a transient infection with an ELISA-positive sample before or after the discordant sample. At least two additional consecutive ELISA-negative samples per child were tested in duplicate by rtPCR for the 16s rRNA gene. A total of 14 of 78 (17.9%) rtPCR reactions were positive, but only 4/78 (5.1%) were positive in both duplicates, representing a total of 3/36 (8.3%) children with an additional rtPCR-positive sample, only one of whom was persistently negative by ELISA. One child with a transient infection had two positive rtPCR reactions despite negative ELISA samples. In H. pylori noninfected or transiently infected children, as determined by stool ELISA, additional ELISA-negative/rtPCR-positive stool samples were found in 8.3% of children, but a possible persistent infection was only identified in 2.7% of children. Thus, the characterization of infection dynamics in children is not being misrepresented by application of stool ELISA. Furthermore, rtPCR does not significantly improve dynamic characterization. © 2016 John Wiley & Sons Ltd.

  17. Comparison of Single-Point and Continuous Sampling Methods for Estimating Residential Indoor Temperature and Humidity.

    Science.gov (United States)

    Johnston, James D; Magnusson, Brianna M; Eggett, Dennis; Collingwood, Scott C; Bernhardt, Scott A

    2015-01-01

    Residential temperature and humidity are associated with multiple health effects. Studies commonly use single-point measures to estimate indoor temperature and humidity exposures, but there is little evidence to support this sampling strategy. This study evaluated the relationship between single-point and continuous monitoring of air temperature, apparent temperature, relative humidity, and absolute humidity over four exposure intervals (5-min, 30-min, 24-hr, and 12-days) in 9 northern Utah homes, from March-June 2012. Three homes were sampled twice, for a total of 12 observation periods. Continuous data-logged sampling was conducted in homes for 2-3 wks, and simultaneous single-point measures (n = 114) were collected using handheld thermo-hygrometers. Time-centered single-point measures were moderately correlated with short-term (30-min) data logger mean air temperature (r = 0.76, β = 0.74), apparent temperature (r = 0.79, β = 0.79), relative humidity (r = 0.70, β = 0.63), and absolute humidity (r = 0.80, β = 0.80). Data logger 12-day means were also moderately correlated with single-point air temperature (r = 0.64, β = 0.43) and apparent temperature (r = 0.64, β = 0.44), but were weakly correlated with single-point relative humidity (r = 0.53, β = 0.35) and absolute humidity (r = 0.52, β = 0.39). Of the single-point RH measures, 59 (51.8%) deviated more than ±5%, 21 (18.4%) deviated more than ±10%, and 6 (5.3%) deviated more than ±15% from data logger 12-day means. Where continuous indoor monitoring is not feasible, single-point sampling strategies should include multiple measures collected at prescribed time points based on local conditions.

  18. Sample Size Induced Brittle-to-Ductile Transition of Single-Crystal Aluminum Nitride

    Science.gov (United States)

    2015-08-01

    Aluminum Nitride by GA Gazonas and JW McCauley Weapons and Materials Research Directorate, ARL JJ Guo, KM Reddy, A Hirata, T Fujita, and MW Chen...Sample Size Induced Brittle-to-Ductile Transition of Single-Crystal Aluminum Nitride 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT...their microscopic structure. In this study, we report a size induced brittle-to-ductile transition in single-crystal aluminum nitride (AlN). When the

  19. Assessing the Validity of Single-item Life Satisfaction Measures: Results from Three Large Samples

    Science.gov (United States)

    Cheung, Felix; Lucas, Richard E.

    2014-01-01

    Purpose The present paper assessed the validity of single-item life satisfaction measures by comparing single-item measures to the Satisfaction with Life Scale (SWLS) - a more psychometrically established measure. Methods Two large samples from Washington (N=13,064) and Oregon (N=2,277) recruited by the Behavioral Risk Factor Surveillance System (BRFSS) and a representative German sample (N=1,312) recruited by the Germany Socio-Economic Panel (GSOEP) were included in the present analyses. Single-item life satisfaction measures and the SWLS were correlated with theoretically relevant variables, such as demographics, subjective health, domain satisfaction, and affect. The correlations between the two life satisfaction measures and these variables were examined to assess the construct validity of single-item life satisfaction measures. Results Consistent across three samples, single-item life satisfaction measures demonstrated substantial degree of criterion validity with the SWLS (zero-order r = 0.62 – 0.64; disattenuated r = 0.78 – 0.80). Patterns of statistical significance for correlations with theoretically relevant variables were the same across single-item measures and the SWLS. Single-item measures did not produce systematically different correlations compared to the SWLS (average difference = 0.001 – 0.005). The average absolute difference in the magnitudes of the correlations produced by single-item measures and the SWLS were very small (average absolute difference = 0.015 −0.042). Conclusions Single-item life satisfaction measures performed very similarly compared to the multiple-item SWLS. Social scientists would get virtually identical answer to substantive questions regardless of which measure they use. PMID:24890827

  20. Assessing the validity of single-item life satisfaction measures: results from three large samples.

    Science.gov (United States)

    Cheung, Felix; Lucas, Richard E

    2014-12-01

    The present paper assessed the validity of single-item life satisfaction measures by comparing single-item measures to the Satisfaction with Life Scale (SWLS)-a more psychometrically established measure. Two large samples from Washington (N = 13,064) and Oregon (N = 2,277) recruited by the Behavioral Risk Factor Surveillance System and a representative German sample (N = 1,312) recruited by the Germany Socio-Economic Panel were included in the present analyses. Single-item life satisfaction measures and the SWLS were correlated with theoretically relevant variables, such as demographics, subjective health, domain satisfaction, and affect. The correlations between the two life satisfaction measures and these variables were examined to assess the construct validity of single-item life satisfaction measures. Consistent across three samples, single-item life satisfaction measures demonstrated substantial degree of criterion validity with the SWLS (zero-order r = 0.62-0.64; disattenuated r = 0.78-0.80). Patterns of statistical significance for correlations with theoretically relevant variables were the same across single-item measures and the SWLS. Single-item measures did not produce systematically different correlations compared to the SWLS (average difference = 0.001-0.005). The average absolute difference in the magnitudes of the correlations produced by single-item measures and the SWLS was very small (average absolute difference = 0.015-0.042). Single-item life satisfaction measures performed very similarly compared to the multiple-item SWLS. Social scientists would get virtually identical answer to substantive questions regardless of which measure they use.

  1. Assessing Children's Report of Stool Consistency: Agreement Between the Pediatric Rome III Questionnaire and the Bristol Stool Scale

    NARCIS (Netherlands)

    Vriesman, Mana H.; Velasco-Benitez, Carlos A.; Ramirez, Carmen R.; Benninga, Marc A.; Di Lorenzo, Carlo; Saps, Miguel

    2017-01-01

    Objectives To assess the agreement between the Questionnaire on Pediatric Gastrointestinal Symptoms-Rome III (QPGS-RIII) and the Bristol Stool Scale (BSS) in evaluating stool consistency and the diagnosis of functional constipation in children. Study design Children aged 8-18 years were asked to

  2. Development of the Brussels Infant and Toddler Stool Scale ('BITSS'): protocol of the study

    NARCIS (Netherlands)

    Vandenplas, Yvan; Szajewska, Hania; Benninga, Marc; Di Lorenzo, Carlo; Dupont, Christophe; Faure, Christophe; Miqdadi, Mohamed; Osatakul, Seksit; Ribes-Konickx, Carmen; Saps, Miguel; Shamir, Raanan; Staiano, Annamaria; Franckx, Johan; Green, Robin; Hegar, Badriul; Lemmens, Roel; Salvatore, Silvia; Vieira, Mario; Verghote, Marc; Xinias, Ioannis

    2017-01-01

    The Bristol Stool Form Scale (BSS) which consists of 7 photographs of different stool forms allows assessment of stool consistency (scale 1 for hard lumps to scale 7 for watery stools), in an objective manner in adults. The BSS is also sometimes used to characterise the stools of infants and young

  3. High-pressure oxygenation of thin-wall YBCO single-domain samples

    International Nuclear Information System (INIS)

    Chaud, X; Savchuk, Y; Sergienko, N; Prikhna, T; Diko, P

    2008-01-01

    The oxygen annealing of ReBCO bulk material, necessary to achieve superconducting properties, usually induces micro- and macro-cracks. This leads to a crack-assisted oxygenation process that allows oxygenating large bulk samples faster than single crystals. But excellent superconducting properties are cancelled by the poor mechanical ones. More progressive oxygenation strategy has been shown to reduce drastically the oxygenation cracks. The problem then arises to keep a reasonable annealing time. The concept of bulk Y123 single-domain samples with thin-wall geometry has been introduced to bypass the inherent limitation due to a slow oxygen diffusion rate. But it is not enough. The use of a high oxygen pressure (16 MPa) enables to speed up further the process. It introduces a displacement in the equilibrium phase diagram towards higher temperatures, i.e., higher diffusion rates, to achieve a given oxygen content in the material. Remarkable results were obtained by applying such a high pressure oxygen annealing process on thin-wall single-domain samples. The trapped field of 16 mm diameter Y123 thin-wall single-domain samples was doubled (0.6T vs 0.3T at 77K) using an annealing time twice shorter (about 3 days). The initial development was made on thin bars. The advantage of thin-wall geometry is that such an annealing can be applied directly to a much larger sample

  4. Preparation and characterization of single crystal samples for high-pressure experiments

    Energy Technology Data Exchange (ETDEWEB)

    Farber, D; Antonangeli, D; Aracne, C; Benterou, J

    2005-10-26

    To date, most research utilizing the diamond anvil cell (DAC) has been conducted with polycrystalline samples, thus the results are limited to addressing average bulk properties. However, experiments on single crystals can yield data on a range of orientation dependent properties such as thermal and electrical conductivity, magnetic susceptibility, elasticity and plasticity. Here we report new procedures to produce extremely high-quality metallic single crystal samples of size compatible with DAC experiments in the Mbar range. So far, we have produced samples of zinc, Al{sub 2}O{sub 3}, cobalt, molybdenum and cerium, and have evaluated the quality of the finished samples with white-light interferometry, synchrotron x-ray diffraction and inelastic x-ray scattering.

  5. Assessing Children's Report of Stool Consistency: Agreement Between the Pediatric Rome III Questionnaire and the Bristol Stool Scale.

    Science.gov (United States)

    Vriesman, Mana H; Velasco-Benitez, Carlos A; Ramirez, Carmen R; Benninga, Marc A; Di Lorenzo, Carlo; Saps, Miguel

    2017-11-01

    To assess the agreement between the Questionnaire on Pediatric Gastrointestinal Symptoms-Rome III (QPGS-RIII) and the Bristol Stool Scale (BSS) in evaluating stool consistency and the diagnosis of functional constipation in children. Children aged 8-18 years were asked to describe their stool consistency in the previous month according to the QPGS-RIII and the BSS. Stool consistency according to both instruments was categorized into 3 categories: "hard," "normal," and "liquid." The children's reported stool consistency using the QPGS-RIII and the BSS were compared, and the intrarater agreement between the 2 instruments was measured using the Cohen kappa coefficient (κ). The diagnosis of functional constipation was based on the Rome III criteria, incorporating the assessment of stool consistency according to the QPGS-RIII and the BSS. A total of 1835 children were included. Only slight agreement existed between the QPGS-RIII and the BSS for assessing stool consistency (κ = .046; P = .022). Significantly more children reported hard stools on the BSS compared to the QPGS-RIII (18.0% vs 7.1%; P = .000). The prevalence of functional constipation was 8.6% using the QPGS-RIII and 9.3% using the BSS (P = .134). Only slight agreement exists between the QPGS-RIII and the BSS in the evaluation of stool consistency in children. Better instruments are needed to assess the consistency of stools with a high degree of reliability, both in research and in the clinical setting. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. First-Glance Diagnosis of Strongyloides stercoralis Autoinfection by Stool Microscopy

    OpenAIRE

    Sing, Andreas; Leitritz, Lorenz; Bogner, Johannes R.; Heesemann, Jürgen

    1999-01-01

    We report a case of autoinfection due to Strongyloides stercoralis in a 27-year-old Ethiopian AIDS patient living in Germany for nearly 3 years. This case was diagnosed on the basis of a single-view field in microscopy of a freshly obtained formalin-fixed stool specimen showing both rhabditiform and filariform larvae. The diagnosis of autoinfection by microscopy is discussed in detail.

  7. Two-step stool aerobic training for smokers.

    Science.gov (United States)

    Gimenez, Manuel; Saavedra, Pedro; Martin, Nieves; Lantarón, Ev M; Polu, Elisabeth; Bach, John Robert

    2014-07-01

    The aim of this study was to analyze subjective, physical, and physiologic responses to a standardized incremental 30-min two-step stool test to create an individualized 45-min maximally intensive two-step stool endurance exercise regimen for home training. This is a longitudinal study on 26 consecutively referred male smokers aged 39-66 yrs. Each performed the two-step stool test on two 15-cm steps at 10, 20, 30, 40, 50, and 60 climbs per minute. Exertional dyspnea, oxygen consumption per unit time, ventilation, respiratory rate, tidal volume, heart rate, capillary oxyhemoglobin saturation, physiologic cost index, and oxygen pulse were recorded and compared with those observed during incremental cycle exercise (30 W per 3 mins). Multivariate analysis for each parameter was undertaken as a mixed model. All subjects attained 60 climbs per minute on the two-step stool test and performed 38-42 mins of two-step stool endurance. All parameters reached 80%-96% of cycle maximum oxygen consumption. The subjects found the two-step stool endurance simple and practical to perform at home. There were no complications. The incremental two-step stool test is a simple, cost-effective way to establish a 45-min maximally intensive endurance exercise training program practical for use in the home.

  8. Stool-based biomarkers of interstitial cystitis/bladder pain syndrome

    Science.gov (United States)

    Braundmeier-Fleming, A.; Russell, Nathan T.; Yang, Wenbin; Nas, Megan Y.; Yaggie, Ryan E.; Berry, Matthew; Bachrach, Laurie; Flury, Sarah C.; Marko, Darlene S.; Bushell, Colleen B.; Welge, Michael E.; White, Bryan A.; Schaeffer, Anthony J.; Klumpp, David J.

    2016-01-01

    Interstitial cystitis/bladder pain syndrome (IC) is associated with significant morbidity, yet underlying mechanisms and diagnostic biomarkers remain unknown. Pelvic organs exhibit neural crosstalk by convergence of visceral sensory pathways, and rodent studies demonstrate distinct bacterial pain phenotypes, suggesting that the microbiome modulates pelvic pain in IC. Stool samples were obtained from female IC patients and healthy controls, and symptom severity was determined by questionnaire. Operational taxonomic units (OTUs) were identified by16S rDNA sequence analysis. Machine learning by Extended Random Forest (ERF) identified OTUs associated with symptom scores. Quantitative PCR of stool DNA with species-specific primer pairs demonstrated significantly reduced levels of E. sinensis, C. aerofaciens, F. prausnitzii, O. splanchnicus, and L. longoviformis in microbiota of IC patients. These species, deficient in IC pelvic pain (DIPP), were further evaluated by Receiver-operator characteristic (ROC) analyses, and DIPP species emerged as potential IC biomarkers. Stool metabolomic studies identified glyceraldehyde as significantly elevated in IC. Metabolomic pathway analysis identified lipid pathways, consistent with predicted metagenome functionality. Together, these findings suggest that DIPP species and metabolites may serve as candidates for novel IC biomarkers in stool. Functional changes in the IC microbiome may also serve as therapeutic targets for treating chronic pelvic pain. PMID:27188581

  9. Real time PCR to detect the environmental faecal contamination by Echinococcus multilocularis from red fox stools.

    Science.gov (United States)

    Knapp, Jenny; Millon, Laurence; Mouzon, Lorane; Umhang, Gérald; Raoul, Francis; Ali, Zeinaba Said; Combes, Benoît; Comte, Sébastien; Gbaguidi-Haore, Houssein; Grenouillet, Frédéric; Giraudoux, Patrick

    2014-03-17

    The oncosphere stage of Echinococcus multilocularis in red fox stools can lead, after ingestion, to the development of alveolar echinococcosis in the intermediate hosts, commonly small mammals and occasionally humans. Monitoring animal infection and environmental contamination is a key issue in public health surveillance. We developed a quantitative real-time PCR technique (qPCR) to detect and quantify E. multilocularis DNA released in fox faeces. A qPCR technique using a hydrolysis probe targeting part of the mitochondrial gene rrnL was assessed on (i) a reference collection of stools from 57 necropsied foxes simultaneously investigated using the segmental sedimentation and counting technique (SSCT) (29 positive for E. multilocularis worms and 28 negative animals for the parasite); (ii) a collection of 114 fox stools sampled in the field: two sets of 50 samples from contrasted endemic regions in France and 14 from an E. multilocularis-free area (Greenland). Of the negative SSCT controls, 26/28 were qPCR-negative and two were weakly positive. Of the positive SSCT foxes, 25/29 samples were found to be positive by qPCR. Of the field samples, qPCR was positive in 21/50 (42%) and 5/48 (10.4%) stools (2 samples inhibited), originating respectively from high and low endemic areas. In faeces, averages of 0.1 pg/μl of DNA in the Jura area and 0.7 pg/μl in the Saône-et-Loire area were detected. All qPCR-positive samples were confirmed by sequencing. The qPCR technique developed here allowed us to quantify environmental E. multilocularis contamination by fox faeces by studying the infectious agent directly. No previous study had performed this test in a one-step reaction. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Single injection 51Cr EDTA plasma clearance determination in children using capillary blood samples

    International Nuclear Information System (INIS)

    Broechner-Mortensen, J.; Christoffersen, J.

    1977-01-01

    The reliability of a determination of the total 51 Cr EDTA plasma clearance (e) (and with it the glomerular filtration rate), by a simplified single injection method (injected dose: 4.5 μCi per kg b.w.) using capillary blood samples (0.2 ml), was investigated in twenty children. Clearance values determined from capillary blood samples did not differ significantly from those measured simultaneously from venous blood samples, the mean ratio+-SD being 1.02+-0.06(n = 10). The reproducibility (total day-to-day variation) of E determined from capillary blood samples was 6.7% in children with decreased renal function (n = 3) and 6.9% in children with normal renal function (n = 7). The present data indicate that the use of capillary blood samples is an accurate and very precise approach for determination of E in children. (Auth.)

  11. Stool Investigations for Colorectal Cancer Screening: From Occult Blood Test to DNA Analysis.

    Science.gov (United States)

    Iannone, Andrea; Losurdo, Giuseppe; Pricci, Maria; Girardi, Bruna; Massaro, Antonio; Principi, Mariabeatrice; Barone, Michele; Ierardi, Enzo; Di Leo, Alfredo

    2016-06-01

    We report an update of current methods for colorectal cancer (CRC) screening based on fecal sample analysis. A systematic review of the literature was performed in MEDLINE, EMBASE, and Science Direct electronic databases. Blood in the stools is the first and most used strategy. Fecal occult blood test (FOBT) and fecal immunochemical test (FIT) are the main methods. Both are economic, easy to perform with high specificity, and low sensitivity. Based on CRC multi-step process with genetic and epigenetic alterations in large bowel cell DNA, single mutations or panels of alterations have been detected. These tests have the advantage of a marked improvement of the sensitivity when compared to fecal blood. However, high costs, poor availability, and correct choice of marker panel represent the major limits. A specific sDNA panel including aberrantly methylated BMP3 and NDRG4 promoter regions, mutant k-ras and β-actin (a reference gene for human DNA quantity), and an immunochemical assay for human hemoglobin has been recently approved by Food and Drug Administration. Novel promising biomarkers for CRC screening are represented by microRNAs (miRNAs), a group of 18-25 nucleotide non-coding RNA molecules that regulate gene expression. Reports on these fecal biomarkers are case-control studies, and each of them evaluates single miRNAs or multi-target panels. On the other hand, some fecal proteins have been studied as possible CRC screening markers, even though they demonstrated poor results. Finally, alterations of estrogen receptor-beta (i.e., dramatic reduction in the early stage of CRC) have been demonstrated in tissue samples. Specific investigations are warranted in order to add further noninvasive markers to the panel of CRC screening tools.

  12. Prevalence of Helicobacter pylori in children by noninvasive stool ...

    African Journals Online (AJOL)

    Prevalence of Helicobacter pylori in children by noninvasive stool Antigen Enzyme Immunoassay. Augustine O. Ebonyi, Emeka Ejeliogu, Stanley T. Odigbo, Martha Omoo Ochoga, Stephen Oguche, Anejo-Okopi A. Joseph ...

  13. Stool Xpert MTB/RIF and urine lipoarabinomannan for the diagnosis of tuberculosis in hospitalized HIV-infected children.

    Science.gov (United States)

    LaCourse, Sylvia M; Pavlinac, Patricia B; Cranmer, Lisa M; Njuguna, Irene N; Mugo, Cyrus; Gatimu, John; Stern, Joshua; Walson, Judd L; Maleche-Obimbo, Elizabeth; Oyugi, Julius; Wamalwa, Dalton; John-Stewart, Grace

    2018-01-02

    Tuberculosis (TB) causes substantial morbidity and mortality in HIV-infected children. Sample collection and the paucibacillary nature of TB in children makes diagnosis challenging. Rapid diagnostic tools using easily obtained specimens are urgently needed. Hospitalized, HIV-infected children aged 12 years or less enrolled in a randomized controlled trial (NCT02063880) comparing urgent to post-stabilization antiretroviral therapy initiation in Kenya underwent TB evaluation. At enrollment, sputum or gastric aspirates were collected for TB culture and Xpert, stool for Xpert, and urine for lipoarabinomannan (LAM). When possible, a second sputum/gastric aspirate for culture was obtained. Stool Xpert and urine LAM performance were compared to reference sputum/gastric aspirate culture. Among 165 HIV-infected children, median age was 24 months [interquartile range (IQR) 13-58], median CD4% was 14.3 (IQR 8.9-22.0%), and 114 (69.5%) had severe immunosuppression. Thirteen (7.9%) children had confirmed TB (positive culture and/or Xpert). Sputum/gastric aspirate Xpert, stool Xpert, and urine LAM sensitivities were 60% [95% confidence interval (CI) 26-88%], 63% (95% CI 25-92%), and 43% (95% CI 10-82%), respectively. Specificity was 98% (95% CI 94-100%) for sputum/gastric aspirate Xpert, 99% (95% CI 95-100%) for stool Xpert, and 91% (95% CI 84-95%) for urine LAM. Stool Xpert and urine LAM sensitivity increased among children with severe immunosuppression [80% (95% CI 28-100) and 60% (95% Cl 15-95%)]. Stool Xpert had similar performance compared with sputum/gastric aspirate Xpert to detect TB. Urine LAM had lower sensitivity and specificity, but increased among children with severe immunosuppression. Stool Xpert and urine LAM can aid rapid detection of TB in HIV-infected children using easily accessible samples.

  14. [Procedure and indications of stool examination in parasitology].

    Science.gov (United States)

    Trabelsi, Sonia; Aouinet, Amira; Khaled, Samira

    2012-06-01

    Intestinal parasites are a public health problem in the world especially in tropical and subtropical countries. Despite the improvement in living standards and healthy conditions, these parasitoses remain relatively frequent in Tunisia. Stool specimen examination keeps the fundamental test for screening and diagnosis. It is to directly search the parasite. Respect for the right procedure of collection of stool is an essential step for the reliability and proper interpretation of results of this examination.

  15. Stool consistency is significantly associated with pain perception.

    Directory of Open Access Journals (Sweden)

    Yukiko Shiro

    Full Text Available Commensal as well as pathogenic bacteria can influence a variety of gut functions, thereby leading to constipation and diarrhea in severe cases. In fact, several researchers have reported evidence supporting the association between stool consistency or constipation and the Gut microbiome (GM composition and dysbiosis. GM influences the human health and disease via the gut-brain axis. We thus hypothesized that the pathogenic bacteria increases pain perception to some extent, which means that there could be an association between stool consistency or constipation and pain perception of healthy subjects.Observational study.The aim of the present study was to investigate the association between stool consistency or constipation and pain perception of healthy subjects.Thirty-eight healthy subjects participated in this study. The participants were assessed on their usual stool form (the Bristol Stool Form Scale: BSFS, constipation (the Cleveland Clinic Constipation score: CCS, degree of obesity, pain perception by mechanical stimulus, cold pain threshold, and a questionnaire on psychological state.The BSFS was significantly and positively associated with pain perception, and showed a significant association with anxiety states. Furthermore, pain perception was significantly associated with anxiety states. However, there were no significant associations between the CCS and any independent variables. In addition, we found that a significant predictor to the pain perception was BSFS. Moreover, there were significant relationships among the psychological states, BSFS and obesity.These results suggest that the stool form is associated with pain perception and anxiety status.

  16. Single- versus multiple-sample method to measure glomerular filtration rate.

    Science.gov (United States)

    Delanaye, Pierre; Flamant, Martin; Dubourg, Laurence; Vidal-Petiot, Emmanuelle; Lemoine, Sandrine; Cavalier, Etienne; Schaeffner, Elke; Ebert, Natalie; Pottel, Hans

    2018-01-08

    There are many different ways to measure glomerular filtration rate (GFR) using various exogenous filtration markers, each having their own strengths and limitations. However, not only the marker, but also the methodology may vary in many ways, including the use of urinary or plasma clearance, and, in the case of plasma clearance, the number of time points used to calculate the area under the concentration-time curve, ranging from only one (Jacobsson method) to eight (or more) blood samples. We collected the results obtained from 5106 plasma clearances (iohexol or 51Cr-ethylenediaminetetraacetic acid (EDTA)) using three to four time points, allowing GFR calculation using the slope-intercept method and the Bröchner-Mortensen correction. For each time point, the Jacobsson formula was applied to obtain the single-sample GFR. We used Bland-Altman plots to determine the accuracy of the Jacobsson method at each time point. The single-sample method showed within 10% concordances with the multiple-sample method of 66.4%, 83.6%, 91.4% and 96.0% at the time points 120, 180, 240 and ≥300 min, respectively. Concordance was poorer at lower GFR levels, and this trend is in parallel with increasing age. Results were similar in males and females. Some discordance was found in the obese subjects. Single-sample GFR is highly concordant with a multiple-sample strategy, except in the low GFR range (<30 mL/min). © The Author 2018. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.

  17. Sampling and single particle analysis for the chemical characterisation of fine atmospheric particulates: A review.

    Science.gov (United States)

    Elmes, Michele; Gasparon, Massimo

    2017-11-01

    To better understand the potential environmental and human health impacts of fine airborne particulate matter (APM), detailed physical and chemical characterisation is required. The only means to accurately distinguish between the multiple compositions in APM is by single particle analysis. A variety of methods and instruments are available, which range from filter-based sample collection for off-line laboratory analysis to on-line instruments that detect the airborne particles and generate size distribution and chemical data in real time. There are many reasons for sampling particulates in the ambient atmosphere and as a consequence, different measurement strategies and sampling devices are used depending on the scientific objectives and subsequent analytical techniques. This review is designed as a guide to some of the techniques available for the sampling and subsequent chemical analysis of individual inorganic particles. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. A simple microfluidic dispenser for single-microparticle and cell samples.

    Science.gov (United States)

    Kasukurti, A; Eggleton, C D; Desai, S A; Disharoon, D I; Marr, D W M

    2014-12-21

    Non-destructive isolation of single-cells has become an important need for many biology research laboratories; however, there is a lack of easily employed and inexpensive tools. Here, we present a single-particle sample delivery approach fabricated from simple, economical components that may address this need. In this, we employ unique flow and timing strategies to bridge the significant force and length scale differences inherent in transitioning from single particle isolation to delivery. Demonstrating this approach, we use an optical trap to isolate individual microparticles and red blood cells that are dispensed within separate 50 μl droplets off a microfluidic chip for collection into microscope slides or microtiter plates.

  19. Prevalence of amoebiasis in a model research community and its confirmation using stool antigen elisa for Entamoeba histolytica.

    Science.gov (United States)

    Akhtar, Tasleem; Khan, Aamir Ghafoor; Ahmed, Israr; Nazli, Rubina; Haider, Jamila

    2016-09-01

    Entamoeba histolytica (E. histolytica) produces an invasive disease called amoebiasis, which commonly produces diarrhea with or without blood in both children and adults, leading to high morbidity and mortality. Entamoeba dispar (E. Dispar) is a non invasive, non pathogenic organism. Both Entamoeba histolytica and Entamoeba Dispar look alike on microscopy and therefore cannot be differentiated unless checked on ELISA, PCR or other specific method. To calculate the actual prevalence of pathogenic amoebiasis in children by comparing the stool microscopy with ELISA stool antigen i.e. gold standard. Across sectional, comparative study. Children under five years in a community village Budhni, District Peshawar. A sample of 288 children aged Entamoeba histolytica. The specificity and sensitivity of microscopic method was calculated against ELISA. Data was analyzed using statistical computer software package SPSS version 10.0. A total of 288 stool specimens were collected and examined for Entamoeba histolytica. Out of these 36(12.5%) stools were positive for E. histolyticaon microscopy while 14(4.9%) were positive on ELISA. Out of 14 ELISA positive samples, 10 samples were also positive on microscopy while 4 were ELISA positive but microscopy negative. About 22 samples, which were positive on microscopy were negative on ELISA indicating that these samples might have been of E. Dispar which is non pathogenic protozoa. The sensitivity and specificity of microscopic method was 71.4% and 90.5% respectively, as against stool antigen test. Actual prevalence of Entamoeba histolytica is low in the area. Stool ELISA was able to differentiate between pathogenic Entamoeba histolytica and the non-pathogenic Entamoeba dispar and thus can minimize unnecessary antiamoebic treatment in these children.

  20. Investigation of applicability of extrapolation method for sample field determination in single-yoke measuring setup

    Czech Academy of Sciences Publication Activity Database

    Stupakov, Oleksandr

    2006-01-01

    Roč. 307, - (2006), s. 279-287 ISSN 0304-8853 R&D Projects: GA AV ČR(CZ) 1QS100100508 Institutional research plan: CEZ:AV0Z10100520 Keywords : magnetic measurement * open magnetic sample * surface field determination * single-yoke setup * magnetic non-destructive testing Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.212, year: 2006

  1. Chromium-51-EDTA clearance in adults with a single-plasma sample.

    Science.gov (United States)

    Mårtensson, J; Groth, S; Rehling, M; Gref, M

    1998-12-01

    In 1996, a committee on renal clearance recommended a mean sojourn time-based methodology for single-sample determination of plasma clearance of 99mTc-diethylenetriamine pentaacetic acid (DTPA) to be used on adults if the patient's glomerular filtration rate (GFR) is suspected to be >30 ml/min. The main purpose of this study was to derive a mean sojourn time-based formula for calculation of 51Cr-ethylenediamine tetraacetic acid (EDTA) clearance in adults. Two groups of patients with 51Cr-EDTA clearance (Cl) between 16 and 172 ml/min were studied. In Group I (n = 46), reference Cl was determined as a multiplasma sample, single-injection method (ClSM). Sixteen blood samples were drawn from 0 until 5 hr after a single intravenous injection of 51Cr-EDTA. In Group II (n = 1046), reference Cl was determined by the Brøchner-Mortensen four-sample clearance method (ClBM). The plasma time-activity curves of Group I were used to derive two mean sojourn time-based formulas (Formulas 1 and 2) for calculation of a single-sample clearance. Formula 1 was derived from the entire time-activity curve, whereas the derivation of Formula 2 used only the final slope of the time-activity curve. The accuracy of the two formulas and the Christensen and Groth 99mTc-DTPA formula was tested on Group II. Chromium-51-EDTA Cl calculated by Formula 1 was almost identical to the Cl calculated by the reference Cl method (r = 0.982; SDdiff = 5.82 ml/min). Both 51Cr-EDTA Cl calculated by Formula 2 and by the 99mTc-DTPA formula showed close correlation with the reference method (r = 0.976, r = 0.985, respectively) but systematically overestimated GFR for the whole range of clearance values by 3.5 and 3.2 ml/min (ptime methodology. The determination is marginally more accurate (ptime-activity curve than from only the final slope. The single-sample formula derived for determination of 99mTc-DTPA Cl tends slightly to overestimate GFR if used to calculate 51Cr-EDTA Cl.

  2. Microwave-assisted headspace single-drop microextration of chlorobenzenes from water samples

    Energy Technology Data Exchange (ETDEWEB)

    Vidal, Lorena [Departamento de Quimica Analitica, Nutricion y Bromatologia, Universidad de Alicante, P.O. Box 99, E-03080 Alicante (Spain); Domini, Claudia E. [Departamento de Quimica Analitica, Nutricion y Bromatologia, Universidad de Alicante, P.O. Box 99, E-03080 Alicante (Spain); Grane, Nuria [Departamento de Quimica Analitica, Nutricion y Bromatologia, Universidad de Alicante, P.O. Box 99, E-03080 Alicante (Spain); Psillakis, Elefteria [Department of Environmental Engineering, Technical University of Crete, Polytechneioupolis, GR-73100 Chania, Crete (Greece); Canals, Antonio [Departamento de Quimica Analitica, Nutricion y Bromatologia, Universidad de Alicante, P.O. Box 99, E-03080 Alicante (Spain)]. E-mail: a.canals@ua.es

    2007-05-29

    A one-step and in-situ sample preparation method used for quantifying chlorobenzene compounds in water samples has been developed, coupling microwave and headspace single-drop microextraction (MW-HS-SDME). The chlorobenzenes in water samples were extracted directly onto an ionic liquid single-drop in headspace mode under the aid of microwave radiation. For optimization, a Plackett-Burman screening design was initially used, followed by a mixed-level factorial design. The factors considered were: drop volume, aqueous sample volume, stirring speed, ionic strength, extraction time, ionic liquid type, microwave power and length of the Y-shaped glass-tube. The optimum experimental conditions found from this statistical evaluation were: a 5 {mu}L microdrop of 1-hexyl-3-methylimidazolium hexafluorophosphate exposed for 20 min to the headspace of a 30 mL aqueous sample, irradiated by microwaves at 200 W and placed in a 50 mL spherical flask connected to a 25 cm Y-shaped glass-tube. Under the optimised experimental conditions, the response of a high performance liquid chromatographic system was found to be linear over the range studied and with correlation coefficients ranging between 0.9995 and 0.9999. The method showed a good level of repeatability, with relative standard deviations varying between 2.3 and 8.3% (n = 5). Detection limits were found in the low {mu}g L{sup -1} range varying between 0.016 and 0.039 {mu}g L{sup -1}. Overall, the performance of the proposed method demonstrated the favourable effect of microwave sample irradiation upon HS-SDME. Finally, recovery studies from different types of environmental water samples revealed that matrix had little effect upon extraction.

  3. Microwave-assisted headspace single-drop microextration of chlorobenzenes from water samples

    International Nuclear Information System (INIS)

    Vidal, Lorena; Domini, Claudia E.; Grane, Nuria; Psillakis, Elefteria; Canals, Antonio

    2007-01-01

    A one-step and in-situ sample preparation method used for quantifying chlorobenzene compounds in water samples has been developed, coupling microwave and headspace single-drop microextraction (MW-HS-SDME). The chlorobenzenes in water samples were extracted directly onto an ionic liquid single-drop in headspace mode under the aid of microwave radiation. For optimization, a Plackett-Burman screening design was initially used, followed by a mixed-level factorial design. The factors considered were: drop volume, aqueous sample volume, stirring speed, ionic strength, extraction time, ionic liquid type, microwave power and length of the Y-shaped glass-tube. The optimum experimental conditions found from this statistical evaluation were: a 5 μL microdrop of 1-hexyl-3-methylimidazolium hexafluorophosphate exposed for 20 min to the headspace of a 30 mL aqueous sample, irradiated by microwaves at 200 W and placed in a 50 mL spherical flask connected to a 25 cm Y-shaped glass-tube. Under the optimised experimental conditions, the response of a high performance liquid chromatographic system was found to be linear over the range studied and with correlation coefficients ranging between 0.9995 and 0.9999. The method showed a good level of repeatability, with relative standard deviations varying between 2.3 and 8.3% (n = 5). Detection limits were found in the low μg L -1 range varying between 0.016 and 0.039 μg L -1 . Overall, the performance of the proposed method demonstrated the favourable effect of microwave sample irradiation upon HS-SDME. Finally, recovery studies from different types of environmental water samples revealed that matrix had little effect upon extraction

  4. Detection of Janus Kinase 2 gene single point mutation in real samples with electrochemical DNA biosensor.

    Science.gov (United States)

    Topkaya, Seda Nur; Kosova, Buket; Ozsoz, Mehmet

    2014-02-15

    Janus Kinase 2 (JAK2) gene single point mutations, which have been reported to be associated with myeloproliferative disorders, are usually detected through conventional methods such as melting curve assays, allele-specific and quantitative Polymerase Chain Reactions (PCRs). Herein, an electrochemical biosensor for the detection of a Guanine (G) to Thymine (T) transversion at nucleotide position 1849 of the JAK2 gene was reported. Due to clinical importance of this mutation, easy and sensitive tests are needed to be developed. Our aim was to design a biosensor system that is capable of detecting the mutation within less than 1h with high sensitivity. For these purposes, an electrochemical sensing system was developed based on detecting hybridization. Hybridization between probe and its target and discrimination of single point mutation was investigated by monitoring guanine oxidation signals observed at +1.0 V with Differential Pulse Voltammetry (DPV) by using synthetic oligonucleotides and Polymerase Chain Reaction (PCR) amplicons. Hybridization between probe and PCR amplicons was also determined with Electrochemical Impedance Spectroscopy (EIS). We successfully detect hybridization first in synthetic samples, and ultimately in real samples involving blood samples from patients as well as additional healthy controls. The limit of detection (S/N=3) was calculated as 44 pmol of target sequence in a 40-μl reaction volume in real samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Simple and Reproducible Sample Preparation for Single-Shot Phosphoproteomics with High Sensitivity

    DEFF Research Database (Denmark)

    Jersie-Christensen, Rosa R.; Sultan, Abida; Olsen, Jesper V

    2016-01-01

    The traditional sample preparation workflow for mass spectrometry (MS)-based phosphoproteomics is time consuming and usually requires multiple steps, e.g., lysis, protein precipitation, reduction, alkylation, digestion, fractionation, and phosphopeptide enrichment. Each step can introduce chemical...... artifacts, in vitro protein and peptide modifications, and contaminations. Those often result in sample loss and affect the sensitivity, dynamic range and accuracy of the mass spectrometric analysis. Here we describe a simple and reproducible phosphoproteomics protocol, where lysis, denaturation, reduction......, and alkylation are performed in a single step, thus reducing sample loss and increasing reproducibility. Moreover, unlike standard cell lysis procedures the cell harvesting is performed at high temperatures (99 °C) and without detergents and subsequent need for protein precipitation. Phosphopeptides are enriched...

  6. A single sample GnRHa stimulation test in the diagnosis of precocious puberty

    Directory of Open Access Journals (Sweden)

    Yazdani Parvin

    2012-07-01

    Full Text Available Abstract Context Gonadotropin-releasing hormone (GnRH has been the standard test for diagnosing central precocious puberty. Because GnRH is no longer available, GnRH analogues (GnRHa are now used. Random LH concentration, measured by the third-generation immunochemiluminometric assay, is a useful screening tool for central precocious puberty. However, GnRHa stimulation test should be considered, when a basal LH measurement is inconclusive. However optimal sampling times for luteinizing hormone (LH have yet to be established. Purpose To determine the appropriate sampling time for LH post leuprolide challenge. Methods A retrospective analysis of multi-sample GnRHa stimulation tests performed in 155 children (aged 1–9 years referred for precocious puberty to Texas Children’s Hospital. After 20 mcg/kg of SQ leuprolide acetate, samples were obtained at 0, 1, 3, and 6 hours. Results Of 71 children with clinical evidence of central precocious puberty, fifty nine children had a peak LH >5 mIU/mL. 52 (88% of these responders had positive responses at 1 hour (95% CI is 80–96%, whereas all 59 children (100% had a peak LH response >5 mIU/mL at 3 hours (95% CI is 94-100%, P = 0.005. Conclusions A single serum LH sample collected 3 hours post GnRHa challenge is the optimal sample to establish the diagnosis of central precocious puberty.

  7. Direct amplification and species determination of microsporidian DNA from stool specimens.

    Science.gov (United States)

    Katzwinkel-Wladarsch, S; Lieb, M; Helse, W; Löscher, T; Rinder, H

    1996-06-01

    Microsporidia are recognized as a major aetiological agent in chronic diarrhoea of immunocompromised patients. Their detection by light microscopy is hampered by the small size of the spores. A simple and rapid DNA extraction method has been developed for the detection of microsporidian DNA by PCR directly from stool specimens. It can be performed at room temperature in a 1.5-ml microcentrifuge tube format in less than 1 hour. The subsequent nested polymerase chain reaction permits the detection of 3-100 spores in a 0.1-g stool sample. The amplification products can be verified and the species Enterocytozoon bieneusi, Encephalitozoon cuniculi and Encephalitozoon (Septata) intestinalis distinguished by a simple restriction endonuclease digest.

  8. Stool microbiota composition is associated with the prospective risk of Plasmodium falciparum infection.

    Science.gov (United States)

    Yooseph, Shibu; Kirkness, Ewen F; Tran, Tuan M; Harkins, Derek M; Jones, Marcus B; Torralba, Manolito G; O'Connell, Elise; Nutman, Thomas B; Doumbo, Safiatou; Doumbo, Ogobara K; Traore, Boubacar; Crompton, Peter D; Nelson, Karen E

    2015-08-22

    In humans it is unknown if the composition of the gut microbiota alters the risk of Plasmodium falciparum infection or the risk of developing febrile malaria once P. falciparum infection is established. Here we collected stool samples from a cohort composed of 195 Malian children and adults just prior to an intense P. falciparum transmission season. We assayed these samples using massively parallel sequencing of the 16S ribosomal RNA gene to identify the composition of the gut bacterial communities in these individuals. During the ensuing 6-month P. falciparum transmission season we examined the relationship between the stool microbiota composition of individuals in this cohort and their prospective risk of both P. falciparum infection and febrile malaria. Consistent with prior studies, stool microbial diversity in the present cohort increased with age, although the overall microbiota profile was distinct from cohorts in other regions of Africa, Asia and North America. Age-adjusted Cox regression analysis revealed a significant association between microbiota composition and the prospective risk of P. falciparum infection; however, no relationship was observed between microbiota composition and the risk of developing febrile malaria once P. falciparum infection was established. These findings underscore the diversity of gut microbiota across geographic regions, and suggest that strategic modulation of gut microbiota composition could decrease the risk of P. falciparum infection in malaria-endemic areas, potentially as an adjunct to partially effective malaria vaccines.

  9. Dipstick for rapid diagnosis of Shigella flexneri 2a in stool.

    Directory of Open Access Journals (Sweden)

    Faridabano Nato

    2007-04-01

    Full Text Available Shigellosis or bacillary dysentery, an acute bloody diarrhoea, is a major public health burden in developing countries. In the absence of prompt and appropriate treatment, the infection is often fatal, particularly in young malnourished children. Here, we describe a new diagnostic test for rapid detection, in stool, at the bedside of patients, of Shigella flexneri 2a, the most predominant agent of the endemic form of the disease.The test is based on the detection of S.flexneri 2a lipopolysaccharide (LPS using serotype 2a-specific monoclonal antibodies coupled to gold particles and displayed on one-step immunochromatographic dipstick. A concentration as low as 20 ng/ml of LPS is detected in distilled water and in reconstituted stools in under 15 minutes. The threshold of detection corresponds to a concentration of 5x10(7 CFU/ml of S. flexneri 2a, which provides an unequivocal positive reaction in three minutes in distilled water and reconstituted stools. The specificity is 100% when tested with a battery of Shigella and unrelated strains, in culture. When tested in Vietnam, on clinical samples, the specificity and sensitivity were 99.2 and 91.5%, respectively. A decrease of the sensitivity during the evaluation on stool samples was observed after five weeks at room temperature and was due to moistening of the dipsticks caused by the humidity of the air during the fifth week of the evaluation. This drawback is now overcome by improving the packaging and providing dipsticks individually wrapped in waterproof bags.This simple dipstick-bases test represents a powerful tool for case management and epidemiological surveys.

  10. Zooplankton diversity analysis through single-gene sequencing of a community sample

    Directory of Open Access Journals (Sweden)

    Nishida Mutsumi

    2009-09-01

    Full Text Available Abstract Background Oceans cover more than 70% of the earth's surface and are critical for the homeostasis of the environment. Among the components of the ocean ecosystem, zooplankton play vital roles in energy and matter transfer through the system. Despite their importance, understanding of zooplankton biodiversity is limited because of their fragile nature, small body size, and the large number of species from various taxonomic phyla. Here we present the results of single-gene zooplankton community analysis using a method that determines a large number of mitochondrial COI gene sequences from a bulk zooplankton sample. This approach will enable us to estimate the species richness of almost the entire zooplankton community. Results A sample was collected from a depth of 721 m to the surface in the western equatorial Pacific off Pohnpei Island, Micronesia, with a plankton net equipped with a 2-m2 mouth opening. A total of 1,336 mitochondrial COI gene sequences were determined from the cDNA library made from the sample. From the determined sequences, the occurrence of 189 species of zooplankton was estimated. BLASTN search results showed high degrees of similarity (>98% between the query and database for 10 species, including holozooplankton and merozooplankton. Conclusion In conjunction with the Census of Marine Zooplankton and Barcode of Life projects, single-gene zooplankton community analysis will be a powerful tool for estimating the species richness of zooplankton communities.

  11. Estimation of technetium 99m mercaptoacetyltriglycine plasma clearance by use of one single plasma sample

    International Nuclear Information System (INIS)

    Mueller-Suur, R.; Magnusson, G.; Karolinska Inst., Stockholm; Bois-Svensson, I.; Jansson, B.

    1991-01-01

    Recent studies have shown that technetium 99m mercaptoacetyltriglycine (MAG-3) is a suitable replacement for iodine 131 or 123 hippurate in gamma-camera renography. Also, the determination of its clearance is of value, since it correlates well with that of hippurate and thus may be an indirect measure of renal plasma flow. In order to simplify the clearance method we developed formulas for the estimation of plasma clearance of MAG-3 based on a single plasma sample and compared them with the multiple sample method based on 7 plasma samples. The correlation to effective renal plasma flow (ERPF) (according to Tauxe's method, using iodine 123 hippurate), which ranged from 75 to 654 ml/min per 1.73 m 2 , was determined in these patients. Using the developed regression equations the error of estimate for the simplified clearance method was acceptably low (18-14 ml/min), when the single plasma sample was taken 44-64 min post-injection. Formulas for different sampling times at 44, 48, 52, 56, 60 and 64 min are given, and we recommend 60 min as optimal, with an error of estimate of 15.5 ml/min. The correlation between the MAG-3 clearances and ERPF was high (r=0.90). Since normal values for MAG-3 clearance are not yet available, transformation to estimated ERPF values by the regression equation (ERPF=1.86xC MAG-3 +4.6) could be of clinical value in order to compare it with the normal values for ERPF given in the literature. (orig.)

  12. Structural and magnetic properties evolution study method using a single ribbon-shaped sample

    Science.gov (United States)

    Moya, Javier A.

    2017-06-01

    A new type of study is presented for magnetic and structural characterization of amorphous or nanocrystalline metallic alloys in ribbon or wire-shaped samples. A single sample is subjecting to successive steps of flash isocurrent heat treatments with increasing duration in time, followed by a rapid cooling, while magneto-electric properties evolution are scanned in situ at room temperature. When one set of isocurrent heat treatments is finished, the annealing current is increased and a new set of isocurrent treatments starts. The properties studied were the saturation magnetization and the coercive field at 50 Hz, magnetic permeability at 100 kHz and electrical resistance from where we also obtained the crystalline fraction. The method was applied on two samples of Finemet-like alloys and the results were analyzed from the perspective of current literature. With the present method it is possible to obtain a general and meticulous understanding of the structural and magnetic evolution of the samples tested, with a considerable saving of time and samples.

  13. Yield of Stool Culture with Isolate Toxin Testing versus a Two-Step Algorithm Including Stool Toxin Testing for Detection of Toxigenic Clostridium difficile▿

    OpenAIRE

    Reller, Megan E.; Lema, Clara A.; Perl, Trish M.; Cai, Mian; Ross, Tracy L.; Speck, Kathleen A.; Carroll, Karen C.

    2007-01-01

    We examined the incremental yield of stool culture (with toxin testing on isolates) versus our two-step algorithm for optimal detection of toxigenic Clostridium difficile. Per the two-step algorithm, stools were screened for C. difficile-associated glutamate dehydrogenase (GDH) antigen and, if positive, tested for toxin by a direct (stool) cell culture cytotoxicity neutralization assay (CCNA). In parallel, stools were cultured for C. difficile and tested for toxin by both indirect (isolate) C...

  14. A Bayesian predictive sample size selection design for single-arm exploratory clinical trials.

    Science.gov (United States)

    Teramukai, Satoshi; Daimon, Takashi; Zohar, Sarah

    2012-12-30

    The aim of an exploratory clinical trial is to determine whether a new intervention is promising for further testing in confirmatory clinical trials. Most exploratory clinical trials are designed as single-arm trials using a binary outcome with or without interim monitoring for early stopping. In this context, we propose a Bayesian adaptive design denoted as predictive sample size selection design (PSSD). The design allows for sample size selection following any planned interim analyses for early stopping of a trial, together with sample size determination before starting the trial. In the PSSD, we determine the sample size using the method proposed by Sambucini (Statistics in Medicine 2008; 27:1199-1224), which adopts a predictive probability criterion with two kinds of prior distributions, that is, an 'analysis prior' used to compute posterior probabilities and a 'design prior' used to obtain prior predictive distributions. In the sample size determination of the PSSD, we provide two sample sizes, that is, N and N(max) , using two types of design priors. At each interim analysis, we calculate the predictive probabilities of achieving a successful result at the end of the trial using the analysis prior in order to stop the trial in case of low or high efficacy (Lee et al., Clinical Trials 2008; 5:93-106), and we select an optimal sample size, that is, either N or N(max) as needed, on the basis of the predictive probabilities. We investigate the operating characteristics through simulation studies, and the PSSD retrospectively applies to a lung cancer clinical trial. (243) Copyright © 2012 John Wiley & Sons, Ltd.

  15. A quantitative polymerase chain reaction assay for rapid detection of 9 pathogens directly from stools of travelers with diarrhea.

    Science.gov (United States)

    Antikainen, Jenni; Kantele, Anu; Pakkanen, Sari H; Lääveri, Tinja; Riutta, Jukka; Vaara, Martti; Kirveskari, Juha

    2013-10-01

    Every year, 80 million tourists traveling to tropical and subtropical areas contract traveler's diarrhea (TD). Forty percent to 80% of cases are caused by bacteria, yet clinical diagnostic tests are available to identify only a few of the strains that cause TD. We aimed to develop a quantitative polymerase chain reaction (qPCR) assay to identify all major pathogens in stool samples. We developed a low-cost, high-throughput, multiplex qPCR assay for simultaneous detection of 9 bacterial pathogens in stool samples: Salmonella, Yersinia, Campylobacter, and Vibrio cholerae, as well as Shigella or enteroinvasive Escherichia coli, enterohemorrhagic E coli, enterotoxigenic E coli (ETEC), enteroaggregative E coli (EAEC), and enteropathogenic E coli (EPEC). The assay was validated using positive (n = 245) and negative (n = 243) control strains, as well as preselected positive and negative stool samples. In addition, stool samples were collected from 96 returning travelers with TD. The findings were compared with those from routine diagnostic tests. The assay detected the bacterial strains with 100% sensitivity and specificity, compared with results from the reference tests. Of all stool samples collected from travelers with TD, EPEC was found in 47%, EAEC in 46%, ETEC in 22%, enterohemorrhagic E coli in 7%, Campylobacter in 6%, Shigella or enteroinvasive E coli in 2%, and Salmonella in 2%. Multiple pathogens were found in 37% of all samples. We developed a low-cost, high-throughput qPCR assay for use in routine diagnostic analysis and research. It detects the pathogenic bacteria most commonly associated with TD in stool samples with 100% sensitivity and specificity, compared with reference methods. The assay requires 4 hours, whereas current detection methods require 1 to 7 days. At least 1 TD pathogen was identified in stool samples from 76% of returning travelers, whereas conventional methods found a pathogen in only 17%. The most commonly detected bacteria were EPEC

  16. Prediction of two-sample 99mTc-diethylene triamine pentaacetic acid plasma clearance from single-sample method

    International Nuclear Information System (INIS)

    Zuo Li; Ma Ying-Chun; Wang Mei; Zhang Chun-Li; Wang Rong-Fu; Wang Hai-Yan

    2005-01-01

    The objective of this study was to develop an equation to predict dual plasma sample method (DPSM) 99m Tc-diethylene triamine pentaacetic acid ( 99m Tc-DTPA) plasma clearance from single plasma sample method (SPSM), and to clarify the condition in which DPSM can be substituted by SPSM in measurement of glomerular filtration rate (GFR). Patients with chronic kidney disease (CKD) were selected. Watson modified Christensen and Groth equation was used to calculate 99m Tc-DTPA plasma clearance by SPSM (sGFR). The equation recommended by the Nephrourology Committee of the Society of Nuclear Medicine was used to calculate 99m Tc-DTPA plasma clearance by DPSM (tGFR) in each patient. The difference between sGFR and tGFR was expressed as percent of the average of these two methods, and tGFR was predicted from sGFR. Plasma creatinine was measured by the kinetic picrate method, and GFR estimated by abbreviated modification of diet in renal disease (MDRD) equation (aGFR) and Cockcroft-Gault equation (cGFR) were evaluated as criteria in selection of DPSM and SPSM. Three hundred and sixty-nine patients with CKD were selected (208 male and 161 female). The average age and body weight were 51.4±15.5 years and 67.2±12.5 kg, respectively. The causes of CKD were glomerular disease, renal arterial stenosis, chronic tubulointerstitial disease, and other causes or causes unknown. The average tGFR was 62.9±36.5 ml/min/1.73 m 2 , ranging from 1-180 ml/min/1.73 m 2 . sGFR was significantly correlated with tGFR (r=0.9194, p 2 ; in contrast, then tGFR was±30 ml/min/1.73 m 2 , the difference was constant (-1.1%, 95% confidence interval -18.3%, 16.1%), and tGFR could be predicted from sGFR using the equation: predicted tGFR (ml/min/1.73 m 2 )=7 4244+0.7318 x sGFR+0.0022 x sGFR 2 (n=299, r 2 =0.9428, p 2 , the diagnostic sensitivity of a cut off value of aGFR=45 ml/min/1.73 m 2 was 91.8%, and recommended as a criterion in the selection of DPSM and SPSM. When GFR ≥30 ml/min/1.73 m 2 , t

  17. Variation in orgasm occurrence by sexual orientation in a sample of U.S. singles.

    Science.gov (United States)

    Garcia, Justin R; Lloyd, Elisabeth A; Wallen, Kim; Fisher, Helen E

    2014-11-01

    Despite recent advances in understanding orgasm variation, little is known about ways in which sexual orientation is associated with men's and women's orgasm occurrence. To assess orgasm occurrence during sexual activity across sexual orientation categories. Data were collected by Internet questionnaire from 6,151 men and women (ages 21-65+ years) as part of a nationally representative sample of single individuals in the United States. Analyses were restricted to a subsample of 2,850 singles (1,497 men, 1,353 women) who had experienced sexual activity in the past 12 months. Participants reported their sex/gender, self-identified sexual orientation (heterosexual, gay/lesbian, bisexual), and what percentage of the time they experience orgasm when having sex with a familiar partner. Mean occurrence rate for experiencing orgasm during sexual activity with a familiar partner was 62.9% among single women and 85.1% among single men, which was significantly different (F1,2848  = 370.6, P sexual orientation: heterosexual men 85.5%, gay men 84.7%, bisexual men 77.6% (F2,1494  = 2.67, P = 0.07, η(2)  = 0.004). For women, however, mean occurrence rate of orgasm varied significantly by sexual orientation: heterosexual women 61.6%, lesbian women 74.7%, bisexual women 58.0% (F2,1350  = 10.95, P sexual orientation, have less predictable, more varied orgasm experiences than do men and that for women, but not men, the likelihood of orgasm varies with sexual orientation. These findings demonstrate the need for further investigations into the comparative sexual experiences and sexual health outcomes of sexual minorities. © 2014 International Society for Sexual Medicine.

  18. MPLEx: a Robust and Universal Protocol for Single-Sample Integrative Proteomic, Metabolomic, and Lipidomic Analyses

    Energy Technology Data Exchange (ETDEWEB)

    Nakayasu, Ernesto S.; Nicora, Carrie D.; Sims, Amy C.; Burnum-Johnson, Kristin E.; Kim, Young-Mo; Kyle, Jennifer E.; Matzke, Melissa M.; Shukla, Anil K.; Chu, Rosalie K.; Schepmoes, Athena A.; Jacobs, Jon M.; Baric, Ralph S.; Webb-Robertson, Bobbie-Jo; Smith, Richard D.; Metz, Thomas O.; Chia, Nicholas

    2016-05-03

    ABSTRACT

    Integrative multi-omics analyses can empower more effective investigation and complete understanding of complex biological systems. Despite recent advances in a range of omics analyses, multi-omic measurements of the same sample are still challenging and current methods have not been well evaluated in terms of reproducibility and broad applicability. Here we adapted a solvent-based method, widely applied for extracting lipids and metabolites, to add proteomics to mass spectrometry-based multi-omics measurements. Themetabolite,protein, andlipidextraction (MPLEx) protocol proved to be robust and applicable to a diverse set of sample types, including cell cultures, microbial communities, and tissues. To illustrate the utility of this protocol, an integrative multi-omics analysis was performed using a lung epithelial cell line infected with Middle East respiratory syndrome coronavirus, which showed the impact of this virus on the host glycolytic pathway and also suggested a role for lipids during infection. The MPLEx method is a simple, fast, and robust protocol that can be applied for integrative multi-omic measurements from diverse sample types (e.g., environmental,in vitro, and clinical).

    IMPORTANCEIn systems biology studies, the integration of multiple omics measurements (i.e., genomics, transcriptomics, proteomics, metabolomics, and lipidomics) has been shown to provide a more complete and informative view of biological pathways. Thus, the prospect of extracting different types of molecules (e.g., DNAs, RNAs, proteins, and metabolites) and performing multiple omics measurements on single samples is very attractive, but such studies are challenging due to the fact that the extraction conditions differ according to the molecule type. Here, we adapted an organic solvent-based extraction method that demonstrated

  19. Single-Tube Flow Cytometry Assay for the Detection of Mature Lymphoid Neoplasms in Paucicellular Samples.

    Science.gov (United States)

    Stacchini, Alessandra; Demurtas, Anna; Aliberti, Sabrina; Barreca, Antonella; Novero, Domenico; Pacchioni, Donatella

    2016-01-01

    Flow cytometry (FC) has become a useful support for cytomorphologic evaluation (CM) of fine-needle aspirates (FNA) and serous cavity effusions (SCE) in cases of suspected non-Hodgkin lymphoma (NHL). FC results may be hampered by the scarce viability and low cellularity of the specimens. We developed a single-tube FC assay (STA) that included 10 antibodies cocktailed in 8-color labeling, a cell viability dye, and a logical gating strategy to detect NHL in hypocellular samples. The results were correlated with CM and confirmed by histologic or molecular data when available. Using the STA, we detected B-type NHL in 31 out of 103 hypocellular samples (81 FNA and 22 SCE). Of these, 8 were not confirmed by CM and 2 were considered to be only suspicious. The FC-negative samples had a final diagnosis of benign/reactive process (42/72), carcinoma (27/72), or Hodgkin lymphoma (3/72). The STA approach allowed obtainment of maximum immunophenotyping data in specimens containing a low number of cells and a large amount of debris. The information obtained by STA can help cytomorphologists not only to recognize but also to exclude malignant lymphomas. © 2016 S. Karger AG, Basel.

  20. Correlation of Clinical Outcomes With Multiplex Molecular Testing of Stool From Children Admitted to Hospital With Gastroenteritis in Botswana.

    Science.gov (United States)

    Pernica, Jeffrey M; Steenhoff, Andrew P; Welch, Henry; Mokomane, Margaret; Quaye, Isaac; Arscott-Mills, Tonya; Mazhani, Loeto; Lechiile, Kwana; Mahony, James; Smieja, Marek; Goldfarb, David M

    2016-09-01

    Diarrheal disease is a leading cause of death for young children. Most pediatric gastroenteritis is caused by viral pathogens; consequently, current recommendations advocate against routine antibacterial therapy if children present without bloody stools. In this prospective cohort study, we enrolled children with severe acute gastroenteritis admitted to hospital in Botswana. Details of presenting history, physical examination, and course in the hospital were recorded. Stool samples were characterized using a 15 pathogen polymerase chain reaction assay. There were 671 participants with a median age of 8.3 months; 77 (11%) had severe acute malnutrition. Only 74 children had bloody stools, of whom 48 (65%) had a detectable bacterial pathogen, compared to 195 of 592 (33%) of those without. There were 26 deaths (3.9%). Covariates associated with death in multivariable logistic regression were the detection of any of Campylobacter/Shigella/enterotoxigenic Escherichia coli (odds ratio [OR] 2.57, 95% confidence interval [CI] 1.07-6.17), severe acute malnutrition (OR 4.34, 95% CI 1.79-10.5), and antibiotic therapy (OR 8.82, 95% CI 2.03-38.2). There was no significant association between bloody stools and death, and the effect of Campylobacter/Shigella/enterotoxigenic E. coli infection on death was not modified by the presence of bloody stools. Detection of bacterial enteropathogens is associated with increased mortality in children in sub-Saharan Africa. Unfortunately, most children with these infections do not have bloody stools, and bloody dysentery was not found to be associated with worse outcomes. Clinical trials evaluating outcomes associated with more aggressive diagnostic strategies in children presenting with severe acute gastroenteritis in sub-Saharan Africa should be undertaken. © The Author 2015. Published by Oxford University Press on behalf of the Pediatric Infectious Diseases Society. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  1. Gatrointestinal parasites associated with the stools of bottle-fed ...

    African Journals Online (AJOL)

    Gatrointestinal parasites associated with the stools of bottle-fed babies in Benin city, Nigeria. MO Okungbowa, FI Okungbowa. Abstract. No Abstract. Global Journal of Pure and Applied Sciences Vol. 13 (3) 2007: pp.415-419. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL ...

  2. Examination of Fingernail Contents and Stool for Ova, Cyst and ...

    African Journals Online (AJOL)

    The parasites identified were Ascaris lumbericoides, Taenia species, Giardia lamblia and Entamoeba histolytica. Of the 101 of the stool specimens examined 59(58.4%) were positive for any one parasite and multiple infections were identified in 17.8% of the positive cases. A. lumbricoides 24(23.8%) was found to be the ...

  3. Comparative evaluation of direct stool smear and Formol-ether ...

    African Journals Online (AJOL)

    Cryptosporidium is a common cause of diarrhoea in patients with Human Immunodeficiency Virus (HIV)/Acquired Immunodeficiency Syndrome (AIDS). Unfortunately this pathogen is not often checked for in Microbiology laboratories because the formol-ether stool concentration method for identification of Cryptosporidium is ...

  4. Estimation of 131J-Jodohippurateclearance by a simplified method using a single plasma sample

    International Nuclear Information System (INIS)

    Botsch, H.; Golde, G.; Kampf, D.

    1980-01-01

    Theoretical volumes calculated from the reciprocal of the plasma concentration of 131 J-Jodohippurate were compared in 95 patients with clearance values calculated by the 2-compartment-method and in 18 patients with conventional PAH-clearance. For estimating Hippurate-clearance from a single blood sampling the most favorable time is 45 min. after injection (r = 0.96; clearance 400/ml/min.: r = 0.98). Clearance values may be derived from the formula: C = 0.4 + 7.26 V - 0.021 x V 2 (V = injected activity/activity per l plasma taken 45 min. after injection). The simplicity, precision and reproducibility of the above mentioned clearance-method is emphasized. (orig.) [de

  5. Single-breath-hold 3-D CINE imaging of the left ventricle using Cartesian sampling.

    Science.gov (United States)

    Wetzl, Jens; Schmidt, Michaela; Pontana, François; Longère, Benjamin; Lugauer, Felix; Maier, Andreas; Hornegger, Joachim; Forman, Christoph

    2018-02-01

    Our objectives were to evaluate a single-breath-hold approach for Cartesian 3-D CINE imaging of the left ventricle with a nearly isotropic resolution of [Formula: see text] and a breath-hold duration of [Formula: see text]19 s against a standard stack of 2-D CINE slices acquired in multiple breath-holds. Validation is performed with data sets from ten healthy volunteers. A Cartesian sampling pattern based on the spiral phyllotaxis and a compressed sensing reconstruction method are proposed to allow 3-D CINE imaging with high acceleration factors. The fully integrated reconstruction uses multiple graphics processing units to speed up the reconstruction. The 2-D CINE and 3-D CINE are compared based on ventricular function parameters, contrast-to-noise ratio and edge sharpness measurements. Visual comparisons of corresponding short-axis slices of 2-D and 3-D CINE show an excellent match, while 3-D CINE also allows reformatting to other orientations. Ventricular function parameters do not significantly differ from values based on 2-D CINE imaging. Reconstruction times are below 4 min. We demonstrate single-breath-hold 3-D CINE imaging in volunteers and three example patient cases, which features fast reconstruction and allows reformatting to arbitrary orientations.

  6. Single cell PCR amplification of diatoms using fresh and preserved samples

    Science.gov (United States)

    Hamilton, Paul B.; Lefebvre, Keely E.; Bull, Roger D.

    2015-01-01

    Single cell Chelex® DNA extraction and nested PCR amplification were used to examine partial gene sequences from natural diatom populations for taxonomic and phylogenetic studies at and above the level of species. DNA was extracted from cells that were either fresh collected or stored in RNAlater. Extractions from Lugol's fixation were also attempted with limited success. Three partial gene sequences (rbcL, 18S, and psbA) were recovered using existing and new primers with a nested or double nested PCR approach with amplification and success rates between 70 and 96%. An rbcL consensus tree grouped morphologically similar specimens and was consistent across the two primary sample treatments: fresh and RNAlater. This tool will greatly enhance the number of microscopic diatom taxa (and potentially other microbes) available for barcoding and phylogenetic studies. The near-term increase in sequence data for diatoms generated via routine single cell extractions and PCR will act as a multiproxy validation of longer-term next generation genomics. PMID:26528252

  7. Brief communication: Is variation in the cranial capacity of the Dmanisi sample too high to be from a single species?

    Science.gov (United States)

    Lee, Sang-Hee

    2005-07-01

    This study uses data resampling to test the null hypothesis that the degree of variation in the cranial capacity of the Dmanisi hominid sample is within the range variation of a single species. The statistical significance of the variation in the Dmanisi sample is examined using simulated distributions based on comparative samples of modern humans, chimpanzees, and gorillas. Results show that it is unlikely to find the maximum difference observed in the Dmanisi sample in distributions of female-female pairs from comparative single-species samples. Given that two sexes are represented, the difference in the Dmanisi sample is not enough to reject the null hypothesis of a single species. Results of this study suggest no compelling reason to invoke multiple taxa to explain variation in the cranial capacity of the Dmanisi hominids. (c) 2004 Wiley-Liss, Inc

  8. Isolation of Escherichia coli Bacteriophages from the Stool of Pediatric Diarrhea Patients in Bangladesh

    OpenAIRE

    Chibani-Chennoufi, Sandra; Sidoti, Josette; Bruttin, Anne; Dillmann, Marie-Lise; Kutter, Elizabeth; Qadri, Firdausi; Sarker, Shafiqul Alam; Brüssow, Harald

    2004-01-01

    A 3-week coliphage survey was conducted in stool samples from 140 Bangladeshi children hospitalized with severe diarrhea. On the Escherichia coli indicator strain K803, all but one phage isolate had 170-kb genomes and the morphology of T4 phage. In spot tests, the individual T4-like phages infected up to 27 out of 40 diarrhea-associated E. coli, representing 22 O serotypes and various virulence factors; only five of them were not infected by any of these new phages. A combination of diagnosti...

  9. Single sample expression-anchored mechanisms predict survival in head and neck cancer.

    Directory of Open Access Journals (Sweden)

    Xinan Yang

    2012-01-01

    Full Text Available Gene expression signatures that are predictive of therapeutic response or prognosis are increasingly useful in clinical care; however, mechanistic (and intuitive interpretation of expression arrays remains an unmet challenge. Additionally, there is surprisingly little gene overlap among distinct clinically validated expression signatures. These "causality challenges" hinder the adoption of signatures as compared to functionally well-characterized single gene biomarkers. To increase the utility of multi-gene signatures in survival studies, we developed a novel approach to generate "personal mechanism signatures" of molecular pathways and functions from gene expression arrays. FAIME, the Functional Analysis of Individual Microarray Expression, computes mechanism scores using rank-weighted gene expression of an individual sample. By comparing head and neck squamous cell carcinoma (HNSCC samples with non-tumor control tissues, the precision and recall of deregulated FAIME-derived mechanisms of pathways and molecular functions are comparable to those produced by conventional cohort-wide methods (e.g. GSEA. The overlap of "Oncogenic FAIME Features of HNSCC" (statistically significant and differentially regulated FAIME-derived genesets representing GO functions or KEGG pathways derived from HNSCC tissue among three distinct HNSCC datasets (pathways:46%, p<0.001 is more significant than the gene overlap (genes:4%. These Oncogenic FAIME Features of HNSCC can accurately discriminate tumors from control tissues in two additional HNSCC datasets (n = 35 and 91, F-accuracy = 100% and 97%, empirical p<0.001, area under the receiver operating characteristic curves = 99% and 92%, and stratify recurrence-free survival in patients from two independent studies (p = 0.0018 and p = 0.032, log-rank. Previous approaches depending on group assignment of individual samples before selecting features or learning a classifier are limited by design to

  10. Evaluation of Rapid stool antigen test for the diagnosis of Helicobacter pylori infection in patients with dyspepsia

    Directory of Open Access Journals (Sweden)

    Salma Khatun

    2016-07-01

    Full Text Available Background and objectives:Several diagnostic assays are used for the detection of Helicobacter pylori infection in suspected peptic ulcer cases. H. pylori stool antigen test is a non-invasive method for the detection of active infection. The present study has evaluated the efficacy of rapid stool antigen test to diagnose H. pylori infection in patients with dyspepsia. Materials and methods: Adult patients with complains of dyspepsia attending the Department of Gastroenterology, Hepatobiliary and Pancreatic Diseases (GHPD of BIRDEM hospital for endoscopy were included. Gastric biopsy, blood and stool samples were obtained from each participant after informed written consent. Rapid urease test (RUT, serum H. pylori immunoglobulin A (IgA and IgG and rapid H. pylori stool antigen (HpSAg tests were performed. Only stool samples were obtained from 31 neonates aged 1 to 30 days as negative control for HpSAg test. Results: A total of 91 adult patients with complain of dyspepsia were included in the study. Out of 91 cases, 17 (18.7% and 74 (81.3% had peptic ulcer and erosion respectively. HpSAg was positive in 63.7% cases compared to 42.9% and 62.6% respectively by RUT and IgA. The rate of HpSAg positivity was significantly higher (p<0.05 in ulcer compared to erosion cases. HpSAg test was positive in all (100% RUT positive cases. Combination of HpSAg test and IgA yielded highest positive result in both ulcer (82.4% and erosion (84% cases. H. pylori IgG was positive in all cases. Conclusion: The study has demonstrated that HpSAg test is an effective and non-invasive diagnostic tool to detect active H. pylori infection in suspected dyspeptic patients. IMC J Med Sci 2016; 10(2: 39-44

  11. Gold-FISH: A correlative approach to microscopic imaging of single microbial cells in environmental samples

    Science.gov (United States)

    Schmidt, Hannes; Seki, David; Woebken, Dagmar; Eickhorst, Thilo

    2017-04-01

    Fluorescence in situ hybridization (FISH) is routinely used for the phylogenetic identification, detection, and quantification of single microbial cells environmental microbiology. Oligonucleotide probes that match the 16S rRNA sequence of target organisms are generally applied and the resulting signals are visualized via fluorescence microscopy. Consequently, the detection of the microbial cells of interest is limited by the resolution and the sensitivity of light microscopy where objects smaller than 0.2 µm can hardly be represented. Visualizing microbial cells at magnifications beyond light microscopy, however, can provide information on the composition and potential complexity of microbial habitats - the actual sites of nutrient cycling in soil and sediments. We present a recently developed technique that combines (1) the phylogenetic identification and detection of individual microorganisms by epifluorescence microscopy, with (2) the in situ localization of gold-labelled target cells on an ultrastructural level by SEM. Based on 16S rRNA targeted in situ hybridization combined with catalyzed reporter deposition, a streptavidin conjugate labeled with a fluorescent dye and nanogold particles is introduced into whole microbial cells. A two-step visualization process including an autometallographic enhancement of nanogold particles then allows for either fluorescence or electron microscopy, or a correlative application thereof. We will present applications of the Gold-FISH protocol to samples of marine sediments, agricultural soils, and plant roots. The detection and enumeration of bacterial cells in soil and sediment samples was comparable to CARD-FISH applications via fluorescence microscopy. Examples of microbe-surface interaction analysis will be presented on the basis of bacteria colonizing the rhizoplane of rice roots. In principle, Gold-FISH can be performed on any material to give a snapshot of microbe-surface interactions and provides a promising tool for

  12. 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

    Directory of Open Access Journals (Sweden)

    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

  13. Dietary Shifts May Trigger Dysbiosis and Mucous Stools in Giant Pandas (Ailuropoda melanoleuca).

    Science.gov (United States)

    Williams, Candace L; Dill-McFarland, Kimberly A; Vandewege, Michael W; Sparks, Darrell L; Willard, Scott T; Kouba, Andrew J; Suen, Garret; Brown, Ashli E

    2016-01-01

    Dietary shifts can result in changes to the gastrointestinal tract (GIT) microbiota, leading to negative outcomes for the host, including inflammation. Giant pandas (Ailuropoda melanoleuca) are physiologically classified as carnivores; however, they consume an herbivorous diet with dramatic seasonal dietary shifts and episodes of chronic GIT distress with symptoms including abdominal pain, loss of appetite and the excretion of mucous stools (mucoids). These episodes adversely affect the overall nutritional and health status of giant pandas. Here, we examined the fecal microbiota of two giant pandas' non-mucoid and mucoid stools and compared these to samples from a previous winter season that had historically few mucoid episodes. To identify the microbiota present, we isolated and sequenced the 16S rRNA using next-generation sequencing. Mucoids occurred following a seasonal feeding switch from predominately bamboo culm (stalk) to leaves. All fecal samples displayed low diversity and were dominated by bacteria in the phyla Firmicutes and to a lesser extent, Proteobacteria. Fecal samples immediately prior to mucoid episodes had lower microbial diversity as compared to mucoids. Mucoids were mostly comprised of common mucosal-associated taxa including Streptococcus and Leuconostoc species, and exhibited increased abundance for bacteria in the family Pasteurellaceae. Taken together, these findings indicate that mucoids may represent an expulsion of the mucosal lining that is driven by changes in diet. We suggest that these occurrences serve to reset their GIT microbiota following changes in bamboo part preference, as giant pandas have retained a carnivorous GIT anatomy while shifting to an herbivorous diet.

  14. A novel multitarget stool DNA test for colorectal cancer screening.

    Science.gov (United States)

    Malik, Pramod

    2016-01-01

    Review of: Imperiale TF, Ransohoff DF, Itzkowitz SH, Levin TR, Lavin P, Lidgard GP, Ahlquist DA, Berger BM. Multitarget stool DNA testing for colorectal-cancer screening. N Engl J Med 2014;370(14):1287-97. This Practice Pearl reviews the results of a prospective, multicenter, cross-sectional clinical study that evaluated the performance of a new multitarget stool DNA (or mt-sDNA) screening test for colorectal cancer (CRC) and compared it with a fecal immunochemical test (FIT) in individuals at average risk for CRC. The potential impact of this test on the future of CRC screening is also discussed in a brief commentary. mt-sDNA testing is a noninvasive screening test designed to detect DNA biomarkers associated with colorectal neoplasia and occult hemoglobin in the stool. The sensitivity of mt-sDNA testing for detection of CRC was 92.3%, compared with 73.8% for FIT (p = 0.002). Sensitivity for detecting advanced precancerous lesions was 42.4% for mt-sDNA testing and 23.8% for FIT (p testing and FIT were 86.6% and 94.9%, respectively (p testing thus may be a first-line screening option for asymptomatic individuals at average risk for CRC who do not want to have a colonoscopy.

  15. Management of stool land revenue in Ghana: A study of the Nkawie ...

    African Journals Online (AJOL)

    Management of stool land revenue in Ghana: A study of the Nkawie and toase stools of the Atwima Nwabiagya District of the Ashanti Region. ... lands, however, has for long time been beset with many problems including indeterminate boundaries of stool lands, poor record keeping which often results in multiple sales and ...

  16. Time of Passage of First Stool in Newborns in a Tertiary Health ...

    African Journals Online (AJOL)

    diapers. For babies who had not passed stool at discharge, the mobile phone number of the parents was used to contact the parents 6 h until they reported passage of first stool. All babies. Time of Passage of First Stool in Newborns in a. Tertiary Health Facility in Southern Nigeria. Philemon E Okoro, Cosmos E Enyindah1.

  17. Area G perimeter surface-soil and single-stage water sampling: Environmental surveillance for fiscal year 95. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    Childs, M.; Conrad, R.

    1997-09-01

    ESH-19 personnel collected soil and single-stage water samples around the perimeter of Area G at Los Alamos National Laboratory (LANL) during FY 95 to characterize possible radionuclide movement out of Area G through surface water and entrained sediment runoff. Soil samples were analyzed for tritium, total uranium, isotopic plutonium, americium-241, and cesium-137. The single-stage water samples were analyzed for tritium and plutonium isotopes. All radiochemical data was compared with analogous samples collected during FY 93 and 94 and reported in LA-12986 and LA-13165-PR. Six surface soils were also submitted for metal analyses. These data were included with similar data generated for soil samples collected during FY 94 and compared with metals in background samples collected at the Area G expansion area.

  18. Poliovirus vaccine strains detected in stool specimens of immunodeficient children in South Africa.

    Science.gov (United States)

    Pavlov, Dobromir N; Van Zyl, Walda B; Kruger, Mariane; Blignaut, Liezl; Grabow, Willie O K; Ehlers, Marthie M

    2006-01-01

    After exposure to the oral poliovirus vaccine (OPV), immunocompetent persons excrete poliovirus (PV) vaccine strains for a limited period. In contrast, immunodeficient individuals remain sometimes chronically infected, and in some cases, PV excretion times as long as 10 years have been reported. During prolonged replication in the human intestine, the PV vaccine strain almost invariably reverts its attenuated character and acquires neurovirulent properties (vaccine-derived PVs, or VDPVs), which resemble wild-type PV strains. The aim of this study was to determine the occurrence of OPV strains in stools of immunodeficient children from a selected area in South Africa, as a first step toward future research on the prevalence and potential health impact of VDPVs. In a period of 1 year, a total of 164 stool samples of HIV-positive children aged 4 months to 8 years were studied for the excretion of OPV strains. In addition, 23 stool samples from healthy immunocompetent children were analyzed after receiving their OPV immunization. By applying a reverse transcription-polymerase chain reaction in combination with a nested PCR, a total of 54 enteroviruses (EVs) were detected in the stool specimens of the immunodeficient children. Using restriction enzyme analysis, 13 PVs were distinguished from 41 nonpolio EVs (NPEVs). A Sabin-specific RT-triplex PCR confirmed the presence of 7 Sabin PV type 1, 4 Sabin PV type 3, and 2 Sabin PV type 2 isolates. The majority of the NPEV group was made up of 7 coxsackievirus B3 (CBV3), 6 echovirus 11 (ECV11), 5 ECV9, and 3 coxsackievirus A6 (CAV6) isolates. According to the results, two of the immunodeficient patients (P023 and P140) who had received their last OPV immunization more than 15 months before (vaccinated at 14 weeks of age) tested positive for Sabin PVs types 3 and 1, respectively. A 5-year-old immunodeficient patient (P052) who had received her last OPV immunization more than 42 months before (vaccinated at 18 months of age

  19. A Novel Stool PCR Test for Helicobacter pylori May Predict Clarithromycin Resistance and Eradication of Infection at a High Rate.

    Science.gov (United States)

    Beckman, Erin; Saracino, Ilaria; Fiorini, Giulia; Clark, Courtney; Slepnev, Vladimir; Patel, Denise; Gomez, Clarissa; Ponaka, Reddy; Elagin, Vecheslav; Vaira, Dino

    2017-08-01

    Clarithromycin-based regimens are commonly used as a first-line therapy for Helicobacter pylori -positive patients; however, resistance to clarithromycin has led to treatment failures. The aim of this study was to evaluate the feasibility of using stool samples to detect the presence of H. pylori DNA while concurrently detecting mutations associated with resistance to clarithromycin. For this purpose, total DNA was extracted from 294 raw stool specimens from H. pylori -positive and -negative patients. TaqMan real-time PCR amplification was used to detect the presence of H. pylori as well as to predict the phenotype of the organism and the related outcome for patients treated with clarithromycin. Clarithromycin resistance was determined upon analysis of the PCR result. Patients were also tested by a urea breath test and were subjected to esophagogastroduodenoscopy, followed by histology, culture, and a rapid urease test, in order to obtain a consensus patient infection status. Of 294 total stool samples, 227 were deemed true positive. The sensitivity of H. pylori detection by PCR was 93.8%. Of 213 true-positive samples that were sequenced, 36.2% showed point mutations associated with clarithromycin resistance (A2142C, A2142G, A2143G). The final correlation of the mutant genotypes as determined by sequencing with the eradication of infection was 86%. We found that Helicobacter pylori DNA can be detected in human stool specimens with high sensitivity and can therefore be used to determine the presence of the bacterium without obtaining a biopsy sample. Moreover, genotypic resistance to clarithromycin can be predicted without obtaining a biopsy sample, facilitating the choice of the right therapeutic approach. Copyright © 2017 American Society for Microbiology.

  20. Sampling

    CERN Document Server

    Thompson, Steven K

    2012-01-01

    Praise for the Second Edition "This book has never had a competitor. It is the only book that takes a broad approach to sampling . . . any good personal statistics library should include a copy of this book." —Technometrics "Well-written . . . an excellent book on an important subject. Highly recommended." —Choice "An ideal reference for scientific researchers and other professionals who use sampling." —Zentralblatt Math Features new developments in the field combined with all aspects of obtaining, interpreting, and using sample data Sampling provides an up-to-date treat

  1. Detection and mapping of Cannabinoids in single hair samples through rapid derivatization- Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

    OpenAIRE

    Beasley, Emma; Francese, Simona; Bassindale, Thomas

    2016-01-01

    The sample preparation method reported in this work has permitted for the first time the application of Matrix Assisted Laser Desorption Ionization Mass Spectrometry Profiling and Imaging (MALDI-MSP and MALDI-MSI) for the detection and mapping of cannabinoids in a single hair sample. MALDI-MSI analysis of hair samples has recently been suggested as an alternative technique to traditional methods of GC-MS and LC-MS due to simpler sample preparation, the ability to detect a narrower time frame ...

  2. Using the Bristol Stool Scale and Parental Report of Stool Consistency as Part of the Rome III Criteria for Functional Constipation in Infants and Toddlers

    NARCIS (Netherlands)

    Koppen, Ilan J. N.; Velasco-Benitez, Carlos A.; Benninga, Marc A.; Di Lorenzo, Carlo; Saps, Miguel

    2016-01-01

    To evaluate among parents of infants and toddlers the agreement between parental report and the Bristol Stool Scale (BSS) in assessing stool consistency and the effect of both methods on determining the prevalence of functional constipation (FC) according to the Rome III criteria. Parents of

  3. Heat-stabilised rice bran consumption by colorectal cancer survivors modulates stool metabolite profiles and metabolic networks: a randomised controlled trial.

    Science.gov (United States)

    Brown, Dustin G; Borresen, Erica C; Brown, Regina J; Ryan, Elizabeth P

    2017-05-01

    Rice bran (RB) consumption has been shown to reduce colorectal cancer (CRC) growth in mice and modify the human stool microbiome. Changes in host and microbial metabolism induced by RB consumption was hypothesised to modulate the stool metabolite profile in favour of promoting gut health and inhibiting CRC growth. The objective was to integrate gut microbial metabolite profiles and identify metabolic pathway networks for CRC chemoprevention using non-targeted metabolomics. In all, nineteen CRC survivors participated in a parallel randomised controlled dietary intervention trial that included daily consumption of study-provided foods with heat-stabilised RB (30 g/d) or no additional ingredient (control). Stool samples were collected at baseline and 4 weeks and analysed using GC-MS and ultra-performance liquid chromatography-MS. Stool metabolomics revealed 93 significantly different metabolites in individuals consuming RB. A 264-fold increase in β-hydroxyisovaleroylcarnitine and 18-fold increase in β-hydroxyisovalerate exemplified changes in leucine, isoleucine and valine metabolism in the RB group. A total of thirty-nine stool metabolites were significantly different between RB and control groups, including increased hesperidin (28-fold) and narirutin (14-fold). Metabolic pathways impacted in the RB group over time included advanced glycation end products, steroids and bile acids. Fatty acid, leucine/valine and vitamin B6 metabolic pathways were increased in RB compared with control. There were 453 metabolites identified in the RB food metabolome, thirty-nine of which were identified in stool from RB consumers. RB consumption favourably modulated the stool metabolome of CRC survivors and these findings suggest the need for continued dietary CRC chemoprevention efforts.

  4. A cytolethal distending toxin gene-based multiplex PCR assay for detection of Campylobacter spp. in stool specimens and comparison with culture method.

    Science.gov (United States)

    Shiramaru, Sachi; Asakura, Masahiro; Inoue, Haruna; Nagita, Akira; Matsuhisa, Akio; Yamasaki, Shinji

    2012-07-01

    In this study, we evaluated the applicability of cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR for the direct detection and identification of Campylobacter jejuni, C. coli and C. fetus from stool specimens of patients with gastroenteritis in comparison to culture methods. A total of 711 stool specimens were examined for the isolation or detection of campylobacters by using Skirrow's selective agar culture plates, a filtration method and the multiplex PCR assay. Forty-one and 36 C. jejuni strains were isolated by culture and filtration methods, respectively. In addition, 2 and 3 C. coli strains were isolated by Skirrow and the filtration methods, respectively. However, when the multiplex PCR was employed, the cdtB genes of C. jejuni and C. coli were detected in 45 and 4 stool samples, respectively, and 9 C. jejuni PCR-positive samples by multiplex PCR were negative by culture method. Sequence analysis of the PCR products obtained from 8 stool specimens from which campylobacters were not isolated by culture method but the sequences exactly matched with that of the cdtB gene of C. jejuni strain 81-176. None of the remaining stool samples which were culture negative for campylobacters produced any amplicon. Stool samples were defined as Campylobacter-positive if detected by any method. The sensitivity of the multiplex PCR was 83%, which was higher than Skirrow (74%) and filtration method (66%). These data indicate that cdtB gene-based multiplex PCR is a rapid and more sensitive method to identify the most important species of Campylobacter for human diseases. (248).

  5. Comparison of microscopy, ELISA, and real-time PCR for detection of Giardia intestinalis in human stool specimens

    Science.gov (United States)

    Beyhan, Yunus Emre; Taş Cengiz, Zeynep

    2017-08-23

    Background/aim: This study included patients who had digestive system complaints between August 2015 and October 2015. The research was designed to compare conventional microscopy with an antigen detection ELISA kit and the TaqMan-based real-time PCR (RT-PCR) technique for detection of Giardia intestinalis in human stool specimens. Materials and methods: Samples were concentrated by formalin-ether sedimentation technique and microscopic examinations were carried out on wet mount slides. A commercially available ELISA kit (Giardia CELISA, Cellabs, Brookvale, Australia) was used for immunoassay. DNA was extracted from fecal samples of about 200 mg using the QIAamp Fast DNA Stool Mini Kit (QIAGEN, Hilden, Germany) and the LightCycler Nano system (Roche Diagnostics, Mannheim, Germany) was used for the TaqMan-based RT-PCR assay. Results: A total of 94 stool samples, 38 of them diagnosed positive (40.4%) and 56 of them diagnosed negative by microscopy, were selected for evaluation by antigen detection and molecular assays. The prevalence of G. intestinalis infection was found as 46.8% (n: 44) and 79.8% (n: 75) by ELISA and RT-PCR, respectively. RT-PCR revealed by far the highest positivity rate compared to the other two methods. The difference between these methods was found to be statistically significant (P PCR, the sensitivity and specificity of microscopy and ELISA were 50.7% and 100% and 53.3% and 79%, respectively. Conclusion: RT-PCR seems to be much more sensitive and beneficial for rapid and accurate diagnosis of G. intestinalis in human stools.

  6. Field testing of collection cards for Cannabis sativa samples with a single hexanucleotide DNA marker.

    Science.gov (United States)

    Allgeier, Lindsay; Hemenway, John; Shirley, Nicholas; LaNier, Tommy; Coyle, Heather Miller

    2011-09-01

    The validity and feasibility of using DNA collection cards in the field for preservation and analysis of Cannabis sativa genotypes were investigated using a highly specific hexanucleotide marker. Collection cards were submitted to the National Marijuana Initiative, which selectively trained and managed the collection of specific types of samples from a variety of participating agencies. Samples collected at seizure sites included fresh marijuana leaf samples, dried "dispensary" samples, U.S. border seizures, and hashish. Using a standardized PCR kit with custom-labeled oligonucleotide primers specific to marijuana, collection cards produced eight genotypes and 13 different alleles, extremely low baselines, and no cross-reactivity with control plant species. Results were produced from all sample types with the exception of hashish. Plant DNA collection cards represent an easily implementable method for the genetic identification and relatedness of C. sativa street and grow site-seized samples with applications for databasing and market disruption. © 2011 American Academy of Forensic Sciences.

  7. Inert gases in a terra sample - Measurements in six grain-size fractions and two single particles from Lunar 20.

    Science.gov (United States)

    Heymann, D.; Lakatos, S.; Walton, J. R.

    1973-01-01

    Review of the results of inert gas measurements performed on six grain-size fractions and two single particles from four samples of Luna 20 material. Presented and discussed data include the inert gas contents, element and isotope systematics, radiation ages, and Ar-36/Ar-40 systematics.

  8. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing

    OpenAIRE

    J. Riba; T. Gleichmann; S. Zimmermann; R. Zengerle; P. Koltay

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1??m and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35?pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20??m in size....

  9. Dietary shift and dysbiosis may trigger mucous stools in giant pandas (Ailuropoda melanoleuca

    Directory of Open Access Journals (Sweden)

    Candace L Williams

    2016-05-01

    Full Text Available Dietary shifts can result in dysbiosis between the host and its gastrointestinal tract (GIT microbiota, leading to negative outcomes including inflammation. Giant pandas (Ailuropoda melanoleuca are physiologically classified as carnivores; however, they consume a herbivorous diet with dramatic seasonal feeding shifts and episodes of chronic GIT distress with symptoms including abdominal pain, loss of appetite and the excretion of mucous stools (mucoids. These episodes adversely affect the overall nutritional and health status of giant pandas. Here, we examined the fecal microbiota of two giant pandas’ normal and mucoid stools and compared these microbiota to baseline samples from a season with historically few episodes. To identify the microbiota present, we isolated and sequenced 16S rRNA using next-generation sequencing. Mucoids occurred following a seasonal feeding switch from predominately bamboo culm (stalk to leaves. All fecal samples displayed low diversity and were dominated by bacterial in the phyla Firmicutes and to a lesser extent, the Proteobacteria. Fecal samples immediately prior to mucoid episodes had lower microbial diversity compared to baseline samples, followed by increased diversity in mucoids. Mucoids were mostly comprised of common mucosal-associated taxa including Streptococcus and Leuconostoc species, and exhibited increased abundance for bacteria in the family Pasteurellaceae. Taken together, these findings indicate that diet-induced intestinal dysbiosis in giant pandas likely results in an expulsion of the mucosal lining in the form of mucoids. We suggest that these occurrences serve to reset their GIT microbiota, as giant pandas have retained a carnivorous GIT anatomy while shifting to an herbivorous diet.

  10. Simple diagnostic approach to childhood fecal retention using the Leech score and Bristol stool form scale in medical practice.

    Science.gov (United States)

    Koh, Hong; Lee, Mi Jung; Kim, Myung Joon; Shin, Jae Il; Chung, Ki Sup

    2010-02-01

    To assess fecal retention, plain abdominal radiography is frequently used to complement the clinical history and physical examination, and three scoring systems have been proposed by Barr, Blethyn, and Leech on a single abdominal radiography. The aim of the present study was to find simple and useful diagnostic tools for an approach to fecal retention by correlation of the three scoring systems with the clinical characteristics. This study included 76 children (5.6-15.4 years, male : female = 33:43) who presented with various gastrointestinal complaints and 20 healthy children from the years 2004-2008. Defecation characteristics, abdominal pain, anorexia and nausea, the Bristol stool form scale, and colonic transit time were studied. Plain abdominal radiographs were independently scored with the three scoring systems by a pediatrician and a radiologist. The k-value of the Leech score (0.912) between two of the observers was higher than the others (Barr 0.870 and Blethyn 0.670), and the correlation coefficients of the Leech scoring system by a pediatrician in relation to the colonic transit time (r = 0.861, P Bristol stool form scale (r = -0.818, P Bristol stool form scale may be simple and useful diagnostic tools for pediatricians to access childhood fecal loading in outpatient clinics along with a thorough clinical history.

  11. Can they or can they not? Nurses' ability to quantify stool in superabsorbent diapers.

    Science.gov (United States)

    Ledbetter, Laura

    2006-08-01

    Estimates of stool output in diapers is not an appropriate guideline to use in determining fluid loss through stool. This study was conducted to determine the accuracy of the ability of the nurses (RNs) and patient care technicians (PCTs) to quantify stool volume in diapers. Size 3 diapers, baby food (green peas), and water were used to simulate combinations of stool and urine and differing degrees of water loss in stool. The results indicated that RNs' and PCTs' assessments of stool volume became less accurate as water loss increased. There were no differences in estimation accuracy between RN and PCTs, and years of experience for RNs or PCTs did not influence accuracy of estimation. It is important to use a holistic approach for determining hydration status in patients, particularly knowledge of signs and symptoms of dehydration.

  12. Single point aerosol sampling: Evaluation of mixing and probe performance in a nuclear stack

    Energy Technology Data Exchange (ETDEWEB)

    Rodgers, J.C.; Fairchild, C.I.; Wood, G.O. [Los Alamos National Laboratory, NM (United States)] [and others

    1995-02-01

    Alternative Reference Methodologies (ARMs) have been developed for sampling of radionuclides from stacks and ducts that differ from the methods required by the U.S. EPA. The EPA methods are prescriptive in selection of sampling locations and in design of sampling probes whereas the alternative methods are performance driven. Tests were conducted in a stack at Los Alamos National Laboratory to demonstrate the efficacy of the ARMs. Coefficients of variation of the velocity tracer gas, and aerosol particle profiles were determined at three sampling locations. Results showed numerical criteria placed upon the coefficients of variation by the ARMs were met at sampling stations located 9 and 14 stack diameters from flow entrance, but not at a location that is 1.5 diameters downstream from the inlet. Experiments were conducted to characterize the transmission of 10 {mu}m aerodynamic equivalent diameter liquid aerosol particles through three types of sampling probes. The transmission ratio (ratio of aerosol concentration at the probe exit plane to the concentration in the free stream) was 107% for a 113 L/min (4-cfm) anisokinetic shrouded probe, but only 20% for an isokinetic probe that follows the EPA requirements. A specially designed isokinetic probe showed a transmission ratio of 63%. The shrouded probe performance would conform to the ARM criteria; however, the isokinetic probes would not.

  13. Prediction of Delayed Colonic Transit Using Bristol Stool Form and Stool Frequency in Eastern Constipated Patients: A Difference From the West.

    Science.gov (United States)

    Jaruvongvanich, Veeravich; Patcharatrakul, Tanisa; Gonlachanvit, Sutep

    2017-10-30

    The correlation between the Bristol stool form scale (BSFS) and colonic transit time (CTT) has been reported in Western populations. Our study aims to study the relationship between BSFS, stool frequency, and CTT in Eastern patients with chronic constipation. A total of 144 chronic functional constipation patients underwent colonic transit study by using radio-opaque markers, anorectal manometry, and balloon expulsion test. Stool diary including stool forms and frequency was recorded. Delayed CTT was defined as the retention of more than 20.0% of radio-opaque markers in the colon on day 5. Twenty-five patients (17.4%) had delayed colonic transit. Mean 5-day BSFS (OR, 0.51; 95% CI, 0.34-0.79; P = 0.021) and stool frequency (OR, 0.60; 95% CI, 0.44-0.83; P = 0.002) were independently associated with delayed CTT by logistic regression analysis. Mean 5-day BSFS (area under the curve [AUC], 0.73; 95% CI, 0.62-0.84; P < 0.001) and stool frequency (AUC, 0.75; 95% CI, 0.63-0.87; P < 0.001) fairly predicted delayed CTT. The optimal mean 5-day BSFS of ≤ 3 provided 68.0% sensitivity, 69.7% specificity, and 69.4% accuracy, and the optimal stool frequency ≤ 2 bowel movements in 5 days provided 64.0% sensitivity, 83.1% specificity, and 84.0% accuracy for predicting delayed CTT. Both stool form and frequency were significantly associated with delayed CTT. Stool frequency ≤ 2 and BSFS 1-3 rather than BSFS 1-2 that was used in the Westerners could be used as surrogate for delayed CTT in Eastern patients with constipation.

  14. Multicenter Evaluation of Clinical Diagnostic Methods for Detection and Isolation of Campylobacter spp. from Stool.

    Science.gov (United States)

    Fitzgerald, Collette; Patrick, Mary; Gonzalez, Anthony; Akin, Joshua; Polage, Christopher R; Wymore, Kate; Gillim-Ross, Laura; Xavier, Karen; Sadlowski, Jennifer; Monahan, Jan; Hurd, Sharon; Dahlberg, Suzanne; Jerris, Robert; Watson, Renee; Santovenia, Monica; Mitchell, David; Harrison, Cassandra; Tobin-D'Angelo, Melissa; DeMartino, Mary; Pentella, Michael; Razeq, Jafar; Leonard, Celere; Jung, Carrianne; Achong-Bowe, Ria; Evans, Yaaqobah; Jain, Damini; Juni, Billie; Leano, Fe; Robinson, Trisha; Smith, Kirk; Gittelman, Rachel M; Garrigan, Charles; Nachamkin, Irving

    2016-05-01

    The use of culture-independent diagnostic tests (CIDTs), such as stool antigen tests, as standalone tests for the detection of Campylobacter in stool is increasing. We conducted a prospective, multicenter study to evaluate the performance of stool antigen CIDTs compared to culture and PCR for Campylobacter detection. Between July and October 2010, we tested 2,767 stool specimens from patients with gastrointestinal illness with the following methods: four types of Campylobacter selective media, four commercial stool antigen assays, and a commercial PCR assay. Illnesses from which specimens were positive by one or more culture media or at least one CIDT and PCR were designated "cases." A total of 95 specimens (3.4%) met the case definition. The stool antigen CIDTs ranged from 79.6% to 87.6% in sensitivity, 95.9 to 99.5% in specificity, and 41.3 to 84.3% in positive predictive value. Culture alone detected 80/89 (89.9% sensitivity) Campylobacter jejuni/Campylobacter coli-positive cases. Of the 209 noncases that were positive by at least one CIDT, only one (0.48%) was positive by all four stool antigen tests, and 73% were positive by just one stool antigen test. The questionable relevance of unconfirmed positive stool antigen CIDT results was supported by the finding that noncases were less likely than cases to have gastrointestinal symptoms. Thus, while the tests were convenient to use, the sensitivity, specificity, and positive predictive value of Campylobacter stool antigen tests were highly variable. Given the relatively low incidence of Campylobacter disease and the generally poor diagnostic test characteristics, this study calls into question the use of commercially available stool antigen CIDTs as standalone tests for direct detection of Campylobacter in stool. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  15. Simple fecal flotation is a superior alternative to guadruple Kato Katz smear examination for the detection of hookworm eggs in human stool

    DEFF Research Database (Denmark)

    Inpankaew, Tawin; Schär, Fabian; Khieu, Virak

    2014-01-01

    parameters of the Kato-Katz (KK) and simple sodium nitrate flotation technique (SNF) in terms of detection and quantification of hookworm eggs, with PCR as an additional reference test in stool, collected as part of a baseline cross-sectional study in Cambodia. METHODS/PRINCIPLE FINDINGS: Fecal samples...

  16. ‘Prevotella ihumii’ sp. nov. and ‘Varibaculum timonense’ sp. nov., two new bacterial species isolated from a fresh human stool specimen

    Directory of Open Access Journals (Sweden)

    E. Guilhot

    2017-07-01

    Full Text Available Here we report the main characteristics of ‘Prevotella ihumii’ sp. nov., CSUR-P3385T and ‘Varibaculum timonense’ sp. nov., CSUR-P3369T isolated in September 2016 from a fresh stool sample of healthy French volunteer woman.

  17. A single lysis solution for the analysis of tissue samples by different proteomic technologies

    DEFF Research Database (Denmark)

    Gromov, P.; Celis, J.E.; Gromova, I.

    2008-01-01

    -based proteomics (reverse-phase lysate arrays or direct antibody arrays), allowing the direct comparison of qualitative and quantitative data yielded by these technologies when applied to the same samples. The usefulness of the CLB1 solution for gel-based proteomics was further established by 2D PAGE analysis...... taken place in molecular biology, cell biology and genomics there is a pressing need to accelerate the translation of basic discoveries into clinical applications. This need, compounded by mounting evidence that cellular model systems are unable to fully recapitulate all biological aspects of human...... dissease, is driving scientists to increasingly use clinically relevant samples for biomarker and target discovery. Tissues are heterogeneous and as a result optimization of sample preparation is critical for generating accurate, representative, and highly reproducible quantitative data. Although a large...

  18. Multiplex polymerase chain reaction for detection of Campylobacter from stool specimen.

    Science.gov (United States)

    Sarkar, S R; Ray, N C; Hossain, M A; Paul, S K; Sarkar, S; Kobayashi, N

    2014-07-01

    This was a study to prospectively evaluate the sensitivity and specificity of multiplex polymerase chain reaction (PCR) to identify Campylobacter. Multiplex polymerase chain reaction (PCR) based on cadF, hipO & asp gene for Campylobacter genus, C. jejuni & C. coli were tested for detection of Campylobacter jejuni & C. coli in naturally infected faecal samples of human. All the samples were subjected to the cultural isolation of organism and biochemical characterization. The samples resulted in the amplification of a DNA fragment of size 400 bp, 500 bp &735 bp in PCR assay. Two hundred faecal samples comprising diarrheal stools, 23(11.5%) could be detected by isolation whereas 24(12.0%) were found positive by PCR. All culture positive cases were positive by PCR and among 01 culture negative case, were positive by PCR. PCR was found to be more sensitive for Campylobacter detection in faecal samples 12.0% as relative to culture isolation which could detect the organism in 11.5% samples. Sensitivity and specificity of PCR were 100% and 99.4% respectively taking Culture as gold standard. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.

  19. Measurement of glomerular filtration rate in adults: accuracy of five single-sample plasma clearance methods

    DEFF Research Database (Denmark)

    Rehling, M; Rabøl, A

    1989-01-01

    After an intravenous injection of a tracer that is removed from the body solely by filtration in the kidneys, the glomerular filtration rate (GFR) can be determined from its plasma clearance. The method requires a great number of blood samples but collection of urine is not needed. In the present...

  20. Determination of burial dose in incompletely bleached fluvial samples using single grains of quartz

    DEFF Research Database (Denmark)

    Thomsen, Kristina Jørkov; Murray, A.S.; Bøtter-Jensen, Lars

    2007-01-01

    -bleached and gamma-irradiated samples. We show that there is a linear relationship between the over-dispersion and the mean dose. Knowing this uncertainty relationship enables us to estimate the burial dose by comparing the predicted uncertainty on the running mean with that calculated from actual dispersion. (c...

  1. Quantitative in-situ TEM nanotensile testing of single crystal Ni facilitated by a new sample preparation approach.

    Science.gov (United States)

    Samaeeaghmiyoni, Vahid; Idrissi, Hosni; Groten, Jonas; Schwaiger, Ruth; Schryvers, Dominique

    2017-03-01

    Twin-jet electro-polishing and Focused Ion Beam (FIB) were combined to produce small size Nickel single crystal specimens for quantitative in-situ nanotensile experiments in the transmission electron microscope. The combination of these techniques allows producing samples with nearly defect-free zones in the centre in contrast to conventional FIB-prepared samples. Since TEM investigations can be performed on the electro-polished samples prior to in-situ TEM straining, specimens with desired crystallographic orientation and initial microstructure can be prepared. The present results reveal a dislocation nucleation-controlled plasticity, in which small loops induced by FIB near the edges of the samples play a central role. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Evaluation of the diagnostic accuracy of 2 immunochromatographic tests detecting campylobacter in stools and their role in campylobacter infection diagnosis.

    Science.gov (United States)

    Bessède, Emilie; Asselineau, Julien; Perez, Paul; Valdenaire, Guillaume; Richer, Olivier; Lehours, Philippe; Mégraud, Francis

    2018-02-07

    The detection of campylobacters in stools is performed essentially by culture, but this technique has a low sensitivity. New detection methods are now available. Among them, immunochromatography tests (ICTs) are very attractive in that they offer a result within 15 minutes. However, previous studies suggest that these tests have a relatively low specificity. The objective of this study was to evaluate the performance of these tests.During the study period, all patients who consulted the emergency units and had a stool culture were included. Their stool samples were tested with 2 ICTs, RIDA®QUICK Campylobacter and ImmunoCardSTAT!Campy. Stools were also tested by a home-made PCR and 2 commercially available ELISAs when one of the ICTs was positive. The composite reference standard (CRS) was defined as positive if the culture was positive or, in case of a negative culture, if the PCR and one of the ELISAs were positive simultaneously.Three hundred and five patients were included. Among the 50 positive specimens with RIDA®QUICK Campylobacter, 47 were considered as true positives by the CRS, corresponding to a positive predictive value (PPV) of 94.0%. Among the 52 positive specimens with ImmunoCardSTAT!Campy, 44 were considered as true positives by the CRS, corresponding to a PPV of 84.6%. The negative predictive values (NPV) were estimated at 94.9% and 92.4% for RIDA®QUICK and ImmunoCardSTAT!Campy, respectively.ICTs appear to be very efficient and allow a very rapid detection of campylobacters which is important to treat early campylobacter infection with an adapted antibiotherapy. Copyright © 2018 American Society for Microbiology.

  3. A proposal for the measurement of Rashba and Dresselhaus spin–orbit interaction strengths in a single sample

    International Nuclear Information System (INIS)

    Maiti, Santanu K.; Sil, Shreekantha; Chakrabarti, Arunava

    2012-01-01

    We establish an exact analytical treatment for the determination of the strengths of the Rashba and Dresselhaus spin–orbit interactions in a single sample by measuring persistent spin current. A hidden symmetry is exploited in the Hamiltonian to show that the spin current vanishes when the strength of the Dresselhaus interaction becomes equal to the strength of the Rashba term. The results are sustained even in the presence of disorder and thus an experiment in this regard will be challenging. -- Highlights: ► An exact analytical treatment is given for the measurement of spin–orbit interaction strengths in a single sample. ► Persistent spin current is calculated. ► Our present analysis gives us confidence to propose an experiment in this line.

  4. A Field-Expedient Method for Direct Detection of Enterotoxigenic E Coli and Shigella from Stool.

    Science.gov (United States)

    Silapong, Sasikorn; Neesanant, Pimmnapar; Sethabutr, Orntipa; Lertsethtakarn, Paphavee; Bodhidatta, Ladaporn; McAvin, James C; Mason, Carl J

    2015-01-01

    We describe a field-expedient analytic system that fills a unique and critical public health role and potentially provides a valuable aid in diagnostics. Dual-fluorigenic, hydrolysis probe (TaqMan), PCR assays for detection of causative agents of enterotoxigenic Escherichia coli (ETEC) disease and shigellosis/bacillary dysentery were prepared in a thermal-stable, hydrolytic enzyme resistant format. The assays were packaged as a kit for use with a portable, ruggedized, qRT-PCR thermocycler. The analytical limit of detection of each q ETEC-STIa, ETEC-STIb, and ETEC-LT assay was 30 colony forming units (CFU) and Shigella/enteroinvasive E coli assay was 3 CFU. During field evaluation, testing was conducted using a blind-panel of 138 stored stool samples previously obtained from enterotoxigenic E coli disease (n=91) and shigellosis (n=47) patients. Sample processing and analyses were completed in 3 days. Test results of the qRT-PCR assays showed promise as aid in pathogen identification when compared to culture, digoxigenin-labeled probe (ETEC), and serotyping (Shigella) the qRT-PCR. The sensitivity of each of the 4 qRT-PCR assays was 100% and specificity was ETEC-STIa (92.4%), ETEC-STIb (92.6%), ETEC-LT (79.6%), and Shigella/enteroinvasive E coli (81.6%). Sequencing of qRT-PCR amplicon indicated that the sensitivity and specificity of each qRT-PCR assay exceeded the comparator methods. The system shows promise as a rapid method for direct detection of ETEC and Shigella from stool and is applicable for use in clinical diagnostics and biosurveillance as an extension of temporary field laboratories or as a part of fixed reference laboratory facilities.

  5. Isolation of RNA from tumor samples: single-step guanidinium acid-phenol method.

    Science.gov (United States)

    Robertson, Naomi; Leek, Russell

    2006-01-01

    The guanidinium acid-phenol method of RNA extraction is relatively fast (4 h) and is useful for the processing of large numbers of samples, without the need for ultracentrifugation. This protocol produces total RNA that includes ribosomal, transfer, and messenger RNA. This high-quality RNA is suitable for Northern blot analysis, dot-blot hybridization, poly (A) RNA selection, in vitro translation, cDNA library construction, reverse transcriptase-polymerase chain reaction, ribonuclease protection assay, and primer extension experiments.

  6. Mathematical inference on helminth egg counts in stool and its applications in mass drug administration programmes to control soil-transmitted helminthiasis in public health.

    Science.gov (United States)

    Levecke, Bruno; Anderson, Roy M; Berkvens, Dirk; Charlier, Johannes; Devleesschauwer, Brecht; Speybroeck, Niko; Vercruysse, Jozef; Van Aelst, Stefan

    2015-03-01

    In the present study, we present a hierarchical model based on faecal egg counts (FECs; expressed in eggs per 1g of stool) in which we first describe the variation in FECs between individuals in a particular population, followed by describing the variance due to counting eggs under a microscope separately for each stool sample. From this general framework, we discuss how to calculate a sample size for assessing a population mean FEC and the impact of an intervention, measured as reduction in FECs, for any scenario of soil-transmitted helminth (STH) epidemiology (the intensity and aggregation of FECs within a population) and diagnostic strategy (amount of stool examined (∼sensitivity of the diagnostic technique) and examination of individual/pooled stool samples) and on how to estimate prevalence of STH in the absence of a gold standard. To give these applications the most wide relevance as possible, we illustrate each of them with hypothetical examples. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Value of Tropheryma whipplei quantitative polymerase chain reaction assay for the diagnosis of Whipple disease: usefulness of saliva and stool specimens for first-line screening.

    Science.gov (United States)

    Fenollar, Florence; Laouira, Sonia; Lepidi, Hubert; Rolain, Jean-Marc; Raoult, Didier

    2008-09-01

    Whipple disease (WD) is a chronic infectious disease caused by Tropheryma whipplei. WD DNA has been found in stool and saliva specimens from patients and asymptomatic carriers. A total of 4418 samples that were sent to our center for determination of WD were tested by a T. whipplei-specific quantitative polymerase chain reaction (PCR) based on repetitive sequences. Definite WD was diagnosed in 71 patients, including 55 patients with classic WD (defined by positive results of periodic acid-Schiff staining and/or specific immunohistochemistry of small-bowel biopsy specimens) and 16 patients with localized WD (including patients with endocarditis, neurologic infection, and uveitis). Of the persons without WD, 2.3% had stool specimens positive for T. whipplei by PCR and 0.2% had saliva specimens positive for T. whipplei by PCR. Diagnosis of WD was likely in patients with positive results of both PCR of saliva specimens and PCR of stool specimens (positive predictive value, 95.2%). When the bacterial load was >10(4) colony-forming units per g of stool, the positive predictive value was 100%. A negative result of PCR of a saliva or stool specimen had a negative predictive value of 99.2% for classic WD. For localized WD, positive results of both PCR of saliva specimens and PCR of stool specimens had a sensitivity of 58% (compared with 94% for classic WD). The positive predictive value of testing of blood, cerebrospinal fluid, and urine specimens was 100% for each, and the positive predictive value for testing of duodenal biopsy specimens was 97.5%. T. whipplei-specific quantitative PCR of saliva and stool specimens should be performed as first-line noninvasive screening for WD. When the results for both types of specimens are positive, diagnosis of classic WD should be highly suspected, especially if a high bacterial load is detected. Because PCR of saliva and stool specimens lacks sensitivity for determination of localized WD, invasive samples should be tested on the

  8. Evaluation of four different systems for extraction of RNA from stool suspensions using MS-2 coliphage as an exogenous control for RT-PCR inhibition.

    Directory of Open Access Journals (Sweden)

    Lester M Shulman

    Full Text Available Knowing when, and to what extent co-extracted inhibitors interfere with molecular RNA diagnostic assays is of utmost importance. The QIAamp Viral RNA Mini Kit (A; MagNA Pure LC2.0 Automatic extractor (B; KingFisher (C; and NucliSENS EasyMag (D RNA extraction systems were evaluated for extraction efficiency and co-purification of inhibitors from stool suspensions. Real-Time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR of MS-2 coliphage spiked into each system's lysis buffer served as an external control for both. Cycle thresholds (Cts of the MS2 were determined for RNA extracted from stool suspensions containing unknown (n = 93 or varying amounts of inhibitors (n = 92. Stool suspensions from the latter group were also used to determine whether MS-2 and enterovirus rRT-PCR inhibitions were correlated. Specifically 23 RNA extracts from stool suspensions were spiked with enterovirus RNA after extraction and 13 of these stool suspension were spiked with intact enterovirus before extraction. MS2 rRT-PCR inhibition varied for RNAs extracted by the different systems. Inhibition was noted in 12 (13.0%, 26 (28.3%, 7 (7.6%, and 7 (7.6% of the first 93 RNA extracts, and 58 (63.0%, 55 (59.8%, 37 (40.2% and 30 (32.6% of the second 92 extracts for A, B, C, and D, respectively. Furthermore, enterovirus rRT-PCR inhibition correlated with MS2 rRT-PCR inhibition for added enterovirus RNA or virus particles. In conclusion, rRT-PCR for MS-2 RNA is a good predictor of inhibition of enterovirus RNA extracted from stool suspensions. EasyMag performed the best, however all four extraction methods were suitable provided that external controls identified problematic samples.

  9. Diagnosis of Helicobacter pylori Infection in Children and Adult by Serological Test and Stool Antigen Test

    Directory of Open Access Journals (Sweden)

    G. Azizi

    2013-10-01

    Full Text Available Background: Helicobacter pylori infection causes chronic gastritis that is related to duodenal ulcer, gastric ulcer, and possibly gastric adenocarcinoma. Noninvasive diagnostic tests consist of the urea breath test, serology, and stool antigen testing. Serodiagnosis of H.pylori infection is inaccurate for children. In order to investigate the immune response to H.pylori in children and adult, we compared anti-H pylori IgG and IgA antibodies with H. pylori antigen (HpSA in the stool. Methods: Serum and stool samples were obtained from 218 children and adult patients with clinical symptom in the range of 4 to 77 years old. Paired results of H. pylori serology (IgG and IgA and HpSA were analyzed by enzyme immunoassay methods. Results: There were 218 paired serology and HpSA results for 39 children (≤17 years and 179 adult (≥18 years. The positivity rate of HpSA (45.8% was significantly lower (P<0.001 than those for H. pylori IgG (54.6% and IgA (28.9%. Moreover in child patients specificity for serological test IgG and IgA were higher than adult. Conclusion: In this study, HpSA was sensitive and specific as a clinical and epidemiological tool to evaluate H. pylori infection. IgG correlated better with HpSA than IgA, and also IgG was much more specific in children than adults confirm the fact that adults are more possible to have been exposed to H. pylori in the past. Using HpSA as the gold standard, we found that the performances of IgG and IgA serology tests differ significantly by age because immature immune response or tolerance to H. pylori is present in childhood and serodiagnosis of H. pylori infection is less useful. Hence, we recommend that laboratories reevaluate reference serologic titers based on age and further clinical correlation is needed to establish the optimal ranges.

  10. Single blood-Hg samples can result in exposure misclassification: temporal monitoring within the Japanese community (United States

    Directory of Open Access Journals (Sweden)

    Tsuchiya Ami

    2012-06-01

    Full Text Available Abstract Background The most prominent non-occupational source of exposure to methylmercury is the consumption of fish. In this study we examine a fish consuming population to determine the extent of temporal exposure and investigate the extent to which single time estimates of methylmercury exposure based on blood-Hg concentration can provide reliable estimates of longer-term average exposure. Methods Blood-mercury levels were obtained from a portion of the Arsenic Mercury Intake Biometric Study (AMIBS cohort. Specifically, 56 Japanese women residing in the Puget Sound area of Washington State, US were sampled on three occasions across a one-year period. Results An average of 135 days separated samples, with mean blood-mercury levels for the visits being 5.1, 6.6 and 5.0 μg/l and geometric means being 2.7, 4.5 and 3.1 μg/l. The blood-mercury levels in this group exceed national averages with geometric means for two of the visits being between the 90th and 95th percentiles of nationally observed levels and the lowest geometric mean being between the 75th and 90th percentile. Group means were not significantly different across sampling periods suggesting that exposure of combined subjects remained relatively constant. Comparing intra-individual results over time did not reveal a strong correlation among visits (r = 0.19, 0.50, 0.63 between 1st and 2nd, 2nd and 3rd, and 1st and 3rd sample results, respectively. In comparing blood-mercury levels across two sampling interval combinations (1st and 2nd, 2nd and 3rd, and 1st and 3rd visits, respectively, 58% (n = 34, 53% (n = 31 and 29% (n = 17 of the individuals had at least a 100% difference in blood-Hg levels. Conclusions Point estimates of blood-mercury, when compared with three sample averages, may not reflect temporal variability and individual exposures estimated on the basis of single blood samples should be treated with caution as indicators of long-term exposure

  11. A method for multiple sequential analyses of macrophage functions using a small single cell sample

    Directory of Open Access Journals (Sweden)

    F.R.F. Nascimento

    2003-09-01

    Full Text Available Microbial pathogens such as bacillus Calmette-Guérin (BCG induce the activation of macrophages. Activated macrophages can be characterized by the increased production of reactive oxygen and nitrogen metabolites, generated via NADPH oxidase and inducible nitric oxide synthase, respectively, and by the increased expression of major histocompatibility complex class II molecules (MHC II. Multiple microassays have been developed to measure these parameters. Usually each assay requires 2-5 x 10(5 cells per well. In some experimental conditions the number of cells is the limiting factor for the phenotypic characterization of macrophages. Here we describe a method whereby this limitation can be circumvented. Using a single 96-well microassay and a very small number of peritoneal cells obtained from C3H/HePas mice, containing as little as <=2 x 10(5 macrophages per well, we determined sequentially the oxidative burst (H2O2, nitric oxide production and MHC II (IAk expression of BCG-activated macrophages. More specifically, with 100 µl of cell suspension it was possible to quantify H2O2 release and nitric oxide production after 1 and 48 h, respectively, and IAk expression after 48 h of cell culture. In addition, this microassay is easy to perform, highly reproducible and more economical.

  12. Estimation of effective renal plasma flow in children by use of a single plasma sample after injection of orthoiodohippurate

    International Nuclear Information System (INIS)

    Tauxe, W.N.; Hagge, W.; Stickler, G.B.

    1975-01-01

    Estimates of effective renal plasma flow (ERPF) in adults have been greatly simplified by the use of recently developed regression equations based on a single plasma concentration of radioiodinated orthoiodohippurate (OIH) obtained 44 min after a single intravenous injection of the material. The paper reports similar equations for the prediction of ERPF in children. Studies were made in 30 children of both sexes, aged 4 to 16 years, with a wide variety of renal diseases. A single intravenous injection of labelled OIH was made and plasma was sampled at 10, 15, 20, 25, 30, 40, 50 and 60 min after injection. Plasma concentration curves were analysed by the conventional two-compartment analysis technique (Sapirstein, Matthews) and ERPF was computed from them. In addition to ERPF calculations, sizes of the two volumes of distribution (V 1 and V 2 ), intercompartmental flow rates and fractional rate constants were also calculated. Surface area (SA) was calculated from the DuBois height-weight regression formula. Reciprocals of the OIH plasma concentration in terms of per cent injected dose per litre were calculated at each sampling time; this reciprocal is designated x. These values at each sampling time were plotted against ERPF and regression equations and residual error (Sy.x) were calculated. The optimal sampling time was found to occur at 53 min and the corresponding linear regression equation was 19.33 + 3.87x with a Sy.x value of 36 ml. This compares with an optimal sampling time in adults of 44 min. The mean initial volume of distribution (V 1 ) was found to be 5.3 litres and the mean V 2 was found to be 6.8 litres. In contrast to findings in the previous study in adults these volumes were found to correlate well with SA. A mean intercompartmental flow rate of 276 ml/min was found. No correlation between this value and clinical diagnosis was found. The relationship of the concentration reciprocal and plasma flow was a linear one instead of the polynomial one

  13. Testing the accuracy of a Bayesian central-dose model for single-grain OSL, using known-age samples

    DEFF Research Database (Denmark)

    Guerin, Guillaume; Combès, Benoit; Lahaye, Christelle

    2015-01-01

    remains a difficult task, given the typically considerable dispersion in equivalent dose values measured by OSL - and the numerous sources of such dispersion in measurements. Here, we present the study of 19 samples for which independent age control is available, and whose ages range from 2 to 46ka. Based......-grain OSL and reference ages is about twice as large for the standard approach (12%) as with the Bayesian model (7%). Statistical tests show that, based on our (limited) dataset, the difference between the two models seems to be significant for samples in the age range 4-46ka. Finally, the influence...... on multi-grain OSL age estimates, these samples are presumed to have been both well-bleached at burial, and unaffected by mixing after deposition. Two ways of estimating single-grain ages are then compared: the standard approach on the one hand, consisting of applying the Central Age Model to De values...

  14. Proton-induced single event upset characterisation of a 1 giga-sample per second analog to digital converter

    International Nuclear Information System (INIS)

    Reed, R.A.; Marshall, P.W.; Carts, M.A.

    1999-01-01

    The SPT7760 is an analog to digital converter that is used in satellite for digital processing. In this paper we describe the characterization and analysis of proton-induced single event upsets (SEU) for the SPT7760 operating at sample rates from 125 Msps (Mega-samples per second) to 1 Gsps. The SEU cross-section has been measured as a function of sample rate for various input levels. The data collected is clearly non-linear for all cases. The data shows that this device has a relative low cross-section for proton-induced SEUs and remains functional at a proton dose of 580 krad (Si). (A.C.)

  15. An iPhone application using a novel stool color detection algorithm for biliary atresia screening.

    Science.gov (United States)

    Hoshino, Eri; Hayashi, Kuniyoshi; Suzuki, Mitsuyoshi; Obatake, Masayuki; Urayama, Kevin Y; Nakano, Satoshi; Taura, Yasuyuki; Nio, Masaki; Takahashi, Osamu

    2017-10-01

    The stool color card has been the primary tool for identifying acholic stools in infants with biliary atresia (BA), in several countries. However, BA stools are not always acholic, as obliteration of the bile duct occurs gradually. This study aims to introduce Baby Poop (Baby unchi in Japanese), a free iPhone application, employing a detection algorithm to capture subtle differences in colors, even with non-acholic BA stools. The application is designed for use by caregivers of infants aged approximately 2 weeks-1 month. Baseline analysis to determine optimal color parameters predicting BA stools was performed using logistic regression (n = 50). Pattern recognition and machine learning processes were performed using 30 BA and 34 non-BA images. Additional 5 BA and 35 non-BA pictures were used to test accuracy. Hue, saturation, and value (HSV) were the preferred parameter for BA stool identification. A sensitivity and specificity were 100% (95% confidence interval 0.48-1.00 and 0.90-1.00, respectively) even among a collection of visually non-acholic, i.e., pigmented BA stools and relatively pale-colored non-BA stools. Results suggest that an iPhone mobile application integrated with a detection algorithm is an effective and convenient modality for early detection of BA, and potentially for other related diseases.

  16. The AlSi10Mg samples produced by selective laser melting: single track, densification, microstructure and mechanical behavior

    International Nuclear Information System (INIS)

    Wei, Pei; Wei, Zhengying; Chen, Zhen; Du, Jun; He, Yuyang; Li, Junfeng; Zhou, Yatong

    2017-01-01

    Highlights: • The thermal behavior of AlSi10Mg molten pool was analyzed. • The SLM-processed sample with a relatively low surface roughness was obtained. • Effects of parameters on surface topography of scan track were investigated. • Effects of parameters on microstructure of parts were investigated. • Optimum processing parameters for AlSi10Mg SLM was obtained. - Abstract: This densification behavior and attendant microstructural characteristics of the selective laser melting (SLM) processed AlSi10Mg alloy affected by the processing parameters were systematically investigated. The samples with a single track were produced by SLM to study the influences of laser power and scanning speed on the surface morphologies of scan tracks. Additionally, the bulk samples were produced to investigate the influence of the laser power, scanning speed, and hatch spacing on the densification level and the resultant microstructure. The experimental results showed that the level of porosity of the SLM-processed samples was significantly governed by energy density of laser beam and the hatch spacing. The tensile properties of SLM-processed samples and the attendant fracture surface can be enhanced by decreasing the level of porosity. The microstructure of SLM-processed samples consists of supersaturated Al-rich cellular structure along with eutectic Al/Si situated at the cellular boundaries. The Si content in the cellular boundaries increases with increasing the laser power and decreasing the scanning speed. The hardness of SLM-processed samples was significantly improved by this fine microstructure compared with the cast samples. Moreover, the hardness of SLM-processed samples at overlaps was lower than the hardness observed at track cores.

  17. The AlSi10Mg samples produced by selective laser melting: single track, densification, microstructure and mechanical behavior

    Energy Technology Data Exchange (ETDEWEB)

    Wei, Pei; Wei, Zhengying, E-mail: zywei@mail.xjtu.edu.cn; Chen, Zhen; Du, Jun; He, Yuyang; Li, Junfeng; Zhou, Yatong

    2017-06-30

    Highlights: • The thermal behavior of AlSi10Mg molten pool was analyzed. • The SLM-processed sample with a relatively low surface roughness was obtained. • Effects of parameters on surface topography of scan track were investigated. • Effects of parameters on microstructure of parts were investigated. • Optimum processing parameters for AlSi10Mg SLM was obtained. - Abstract: This densification behavior and attendant microstructural characteristics of the selective laser melting (SLM) processed AlSi10Mg alloy affected by the processing parameters were systematically investigated. The samples with a single track were produced by SLM to study the influences of laser power and scanning speed on the surface morphologies of scan tracks. Additionally, the bulk samples were produced to investigate the influence of the laser power, scanning speed, and hatch spacing on the densification level and the resultant microstructure. The experimental results showed that the level of porosity of the SLM-processed samples was significantly governed by energy density of laser beam and the hatch spacing. The tensile properties of SLM-processed samples and the attendant fracture surface can be enhanced by decreasing the level of porosity. The microstructure of SLM-processed samples consists of supersaturated Al-rich cellular structure along with eutectic Al/Si situated at the cellular boundaries. The Si content in the cellular boundaries increases with increasing the laser power and decreasing the scanning speed. The hardness of SLM-processed samples was significantly improved by this fine microstructure compared with the cast samples. Moreover, the hardness of SLM-processed samples at overlaps was lower than the hardness observed at track cores.

  18. Gene Xpert for Direct Detection of Mycobacterium Tuberculosis in Stool Specimens from Children with Presumptive Pulmonary Tuberculosis.

    Science.gov (United States)

    Moussa, Husseiny Sh; Bayoumi, Faten Sayed; Mohamed, Ahmed Mohamed Ali

    2016-01-01

    Gene Xpert(GX) is a novel real time polymerase chain reaction (RT-PCR) assay which was endorsed by the World Health Organization (WHO) in 2011 for tuberculosis (TB) diagnosis and susceptibility to refampicin(RIF). To evaluate GX for direct diagnosis of TB in stool samples from children with suspected pulmonary Tuberculosis (PTB). Children older than one year and younger than 16 years with presumptive PTB were enrolled and classified to five clinical categories based on clinical, radiological, and laboratory findings: confirmed TB, probable TB, possible TB, Unlikely TB, and not TB. Two stool samples were collected from each child and tested for the presence of Mycobacterium tuberculosis (MTB) by GX and the obtained results were compared to Lowenstien-Jensen (LJ) culture as a gold standard. In total, 115 children were enrolled. 36 had been confirmed with TB, 61 probably TB, 10 possible TB, 5 unlikely TB, and 3 not TB. GX had a sensitivity of 83.33 and 80.56 % and specificity of 98.73 and 99.36 % by patients and samples respectively. GX was positive in 83.3% of confirmed TB as well as 1.6 and 0.8% of probable TB cases by patients and samples respectively. GX provided timely results with quit acceptable sensitivity and good specificity compared to LJ culture. In this study, sensitivity calculations take into account only children with confirmed TB. GX could not detect TB in children with probable TB, so it should not be used alone for TB diagnosis. Further studies for GX stool protocol optimization and assessment is required. © 2016 by the Association of Clinical Scientists, Inc.

  19. Sequence-specific detection of different strains of LCMV in a single sample using tentacle probes.

    Science.gov (United States)

    Franco, Lina S; Holechek, Susan A; Caplan, Michael R; Blattman, Joseph N

    2017-10-13

    Virus infections often result in quasispecies of viral strains that can have dramatic impacts on disease outcomes. However, sequencing of viruses to determine strain composition is time consuming and often cost-prohibitive. Rapid, cost-effective methods are needed for accurate measurement of virus diversity to understand virus evolution and can be useful for experimental systems. We have developed a novel molecular method for sequence-specific detection of RNA virus genetic variants called Tentacle Probes. The probes are modified molecular beacons that have dramatically improved false positive rates and specificity in routine qPCR. To validate this approach, we have designed Tentacle Probes for two different strains of Lymphocytic Choriomeningitis Virus (LCMV) that differ by only 3 nucleotide substitutions, the parental Armstrong and the more virulent Clone-13 strain. One of these mutations is a missense mutation in the receptor protein GP1 that leads to the Armstrong strain to cause an acute infection and Clone-13 to cause a chronic infection instead. The probes were designed using thermodynamic calculations for hybridization between target or non-target sequences and the probe. Using this approach, we were able to distinguish these two strains of LCMV individually by a single nucleotide mutation. The assay showed high reproducibility among different concentrations of viral cDNA, as well as high specificity and sensitivity, especially for the Clone-13 Tentacle Probe. Furthermore, in virus mixing experiments we were able to detect less than 10% of Clone-13 cDNA diluted in Armstrong cDNA. Thus, we have developed a fast, cost-effective approach for identifying Clone-13 strain in a mix of other LCMV strains.

  20. An in vitro tag-and-modify protein sample generation method for single-molecule fluorescence resonance energy transfer.

    Science.gov (United States)

    Hamadani, Kambiz M; Howe, Jesse; Jensen, Madeleine K; Wu, Peng; Cate, Jamie H D; Marqusee, Susan

    2017-09-22

    Biomolecular systems exhibit many dynamic and biologically relevant properties, such as conformational fluctuations, multistep catalysis, transient interactions, folding, and allosteric structural transitions. These properties are challenging to detect and engineer using standard ensemble-based techniques. To address this drawback, single-molecule methods offer a way to access conformational distributions, transient states, and asynchronous dynamics inaccessible to these standard techniques. Fluorescence-based single-molecule approaches are parallelizable and compatible with multiplexed detection; to date, however, they have remained limited to serial screens of small protein libraries. This stems from the current absence of methods for generating either individual dual-labeled protein samples at high throughputs or protein libraries compatible with multiplexed screening platforms. Here, we demonstrate that by combining purified and reconstituted in vitro translation, quantitative unnatural amino acid incorporation via AUG codon reassignment, and copper-catalyzed azide-alkyne cycloaddition, we can overcome these challenges for target proteins that are, or can be, methionine-depleted. We present an in vitro parallelizable approach that does not require laborious target-specific purification to generate dual-labeled proteins and ribosome-nascent chain libraries suitable for single-molecule FRET-based conformational phenotyping. We demonstrate the power of this approach by tracking the effects of mutations, C-terminal extensions, and ribosomal tethering on the structure and stability of three protein model systems: barnase, spectrin, and T4 lysozyme. Importantly, dual-labeled ribosome-nascent chain libraries enable single-molecule co-localization of genotypes with phenotypes, are well suited for multiplexed single-molecule screening of protein libraries, and should enable the in vitro directed evolution of proteins with designer single-molecule conformational

  1. A review of single-sample-based models and other approaches for radiocarbon dating of dissolved inorganic carbon in groundwater

    Science.gov (United States)

    Han, L. F; Plummer, Niel

    2016-01-01

    Numerous methods have been proposed to estimate the pre-nuclear-detonation 14C content of dissolved inorganic carbon (DIC) recharged to groundwater that has been corrected/adjusted for geochemical processes in the absence of radioactive decay (14C0) - a quantity that is essential for estimation of radiocarbon age of DIC in groundwater. The models/approaches most commonly used are grouped as follows: (1) single-sample-based models, (2) a statistical approach based on the observed (curved) relationship between 14C and δ13C data for the aquifer, and (3) the geochemical mass-balance approach that constructs adjustment models accounting for all the geochemical reactions known to occur along a groundwater flow path. This review discusses first the geochemical processes behind each of the single-sample-based models, followed by discussions of the statistical approach and the geochemical mass-balance approach. Finally, the applications, advantages and limitations of the three groups of models/approaches are discussed.The single-sample-based models constitute the prevailing use of 14C data in hydrogeology and hydrological studies. This is in part because the models are applied to an individual water sample to estimate the 14C age, therefore the measurement data are easily available. These models have been shown to provide realistic radiocarbon ages in many studies. However, they usually are limited to simple carbonate aquifers and selection of model may have significant effects on 14C0 often resulting in a wide range of estimates of 14C ages.Of the single-sample-based models, four are recommended for the estimation of 14C0 of DIC in groundwater: Pearson's model, (Ingerson and Pearson, 1964; Pearson and White, 1967), Han & Plummer's model (Han and Plummer, 2013), the IAEA model (Gonfiantini, 1972; Salem et al., 1980), and Oeschger's model (Geyh, 2000). These four models include all processes considered in single-sample-based models, and can be used in different ranges of

  2. Repeat Rifaximin for Irritable Bowel Syndrome: No Clinically Significant Changes in Stool Microbial Antibiotic Sensitivity.

    Science.gov (United States)

    Pimentel, M; Cash, B D; Lembo, A; Wolf, R A; Israel, R J; Schoenfeld, P

    2017-09-01

    Rifaximin has demonstrated efficacy and safety for diarrhea-predominant irritable bowel syndrome (IBS-D). To determine the rifaximin repeat treatment effect on fecal bacterial antibiotic susceptibility. Patients with IBS in Trial 3 (TARGET 3) study who responded to open-label rifaximin 550 mg three times daily for 2 weeks, with symptom recurrence within 18 weeks, were randomized to double-blind treatment: two 2-week repeat courses of rifaximin or placebo, separated by 10 weeks. Prospective stool sample collection occurred before and after open-label rifaximin, before and after the first repeat course, and at the end of the study. Susceptibility testing was performed with 11 antibiotics, including rifaximin and rifampin, using broth microdilution or agar dilution methods. Of 103 patients receiving open-label rifaximin, 73 received double-blind rifaximin (n = 37) or placebo (n = 36). A total of 1429 bacterial and yeast isolates were identified, of which Bacteroidaceae (36.7%) and Enterobacteriaceae (33.9%) were the most common. In the double-blind phase, Clostridium difficile was highly susceptible to rifaximin [minimum inhibitory concentration (MIC) range 0.008-1 µg/mL] and rifampin (MIC range 0.004-0.25 µg/mL). Following double-blind rifaximin treatment, Staphylococcus isolates remained susceptible to rifaximin at all visits (MIC 50 range ≤0.06-32 µg/mL). Rifaximin exposure was not associated with long-term cross-resistance of Bacteroidaceae, Enterobacteriaceae, and Enterococcaceae to rifampin or nonrifamycin antibiotics tested. In this study, short-term repeat treatment with rifaximin has no apparent long-term effect on stool microbial susceptibility to rifaximin, rifampin, and nonrifamycin antibiotics. CLINICALTRIALS. NCT01543178.

  3. A safety equipment list for rotary mode core sampling systems operation in single shell flammable gas tanks

    International Nuclear Information System (INIS)

    SMALLEY, J.L.

    1999-01-01

    This document identifies all interim safety equipment to be used for rotary mode core sampling of single-shell flammable gas tanks utilizing Rotary Mode Core Sampling systems (RMCS). This document provides the safety equipment for RMCS trucks HO-68K-4600, HO-68K-4647, trucks three and four respectively, and associated equipment. It is not intended to replace or supersede WHC-SD-WM-SEL-023, (Kelly 1991), or WHC-SD-WM-SEL-032, (Corbett 1994), which classifies 80-68K-4344 and HO-68K-4345 respectively. The term ''safety equipment'' refers to safety class (SC) and safety significant (SS) equipment, where equipment refers to structures, systems and components (SSC's). The identification of safety equipment in this document is based on the credited design safety features and analysis contained in the Authorization Basis (AB) for rotary mode core sampling operations in single-shell flammable gas tanks. This is an interim safety classification since the AB is interim. This document will be updated to reflect the final RMCS equipment safety classification designations upon completion of a final AB which will be implemented with the release of the Final Safety Analysis Report (FSAR)

  4. Frequency of single nucleotide polymorphisms of some immune response genes in a population sample from São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Léa Campos de Oliveira

    2011-09-01

    Full Text Available Objective: To present the frequency of single nucleotide polymorphismsof a few immune response genes in a population sample from SãoPaulo City (SP, Brazil. Methods: Data on allele frequencies ofknown polymorphisms of innate and acquired immunity genes werepresented, the majority with proven impact on gene function. Datawere gathered from a sample of healthy individuals, non-HLA identicalsiblings of bone marrow transplant recipients from the Hospital dasClínicas da Faculdade de Medicina da Universidade de São Paulo,obtained between 1998 and 2005. The number of samples variedfor each single nucleotide polymorphism analyzed by polymerasechain reaction followed by restriction enzyme cleavage. Results:Allele and genotype distribution of 41 different gene polymorphisms,mostly cytokines, but also including other immune response genes,were presented. Conclusion: We believe that the data presentedhere can be of great value for case-control studies, to define whichpolymorphisms are present in biologically relevant frequencies and toassess targets for therapeutic intervention in polygenic diseases witha component of immune and inflammatory responses.

  5. Incidence of Giardia lamblia Subspecies by PCR-RFLP in Stool Specimens of Hospitalized Children at Urmia Mutahhari Hospital, West Azerbaijan Province, Iran.

    Directory of Open Access Journals (Sweden)

    Khosro Hazrati Tappeh

    2014-12-01

    Full Text Available Giardia lamblia is one of the most prevalent intestinal flagellate protozoa that infects a wide range of vertebrate hosts causing severe intestinal disorder in children.This study was performed to determine subspecies of G.lamblia by the PCR-RFLP method, targeting the glutamate dehydrogenase(gdhlocus, in hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province,Iran and determining the infection transformational storages in this area.Overall, 720 stool specimens were collected from the hospitalized children, 34 samples were positive and Giardia cysts were detected under the microscope. Cysts were partially purified by the sucrose density gradient method and then washed with sterile distilled water to remove effectively the PCR inhibitors. Genomic DNA of G. lamblia isolates was extracted by freeze-thaw cycles followed by phenol/ chloroform/isoamyl alcohol method. The single step PCR-RFLP assay was used to differentiate the assemblages between A and B, which were found in humans. In this method, 432 bp expected size was amplified, and then for detection of subspecies, specific restriction RsaI and BspLI enzymes were used.Totally 34 samples were positive in terms of Giardia cyst out of 720 examined samples microscopically, so the parasite spread rate is reported 4.72%. Analysis PCR-RFLP on these samples revealed that 28 samples (93.3% have the genotype BIII and 2 samples (6.7% belong to the subgroup BIV.PCR-RFLP is a proper analytical method for determining the genotype among parasite types, using the glutamate dehydrogenizes zone's genes. Based on the results, an animal origin of infection cycle is suggested.

  6. Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identification of Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review.

    Science.gov (United States)

    Laude, A; Valot, S; Desoubeaux, G; Argy, N; Nourrisson, C; Pomares, C; Machouart, M; Le Govic, Y; Dalle, F; Botterel, F; Bourgeois, N; Cateau, E; Leterrier, M; Le Pape, P; Morio, F

    2016-02-01

    Microscopy is the reference standard for routine laboratory diagnosis in faecal parasitology but there is growing interest in alternative methods to overcome the limitations of microscopic examination, which is time-consuming and highly dependent on an operator's skills and expertise. Compared with microscopy, DNA detection by PCR is simple and can offer a better turnaround time. However, PCR performances remain difficult to assess as most studies have been conducted on a limited number of positive clinical samples and used in-house PCR methods. Our aim was to evaluate a new multiplex PCR assay (G-DiaParaTrio; Diagenode Diagnostics), targeting Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica. To minimize the turnaround time, PCR was coupled with automated DNA extraction (QiaSymphony; Qiagen). The PCR assay was evaluated using a reference panel of 185 samples established by routine microscopic examination using a standardized protocol including Ziehl-Neelsen staining and adhesin detection by ELISA (E. histolytica II; TechLab). This panel, collected from 12 French parasitology laboratories, included 135 positive samples for G. intestinalis (n = 38), C. parvum/C. hominis (n = 26), E. histolytica (n = 5), 21 other gastrointestinal parasites, together with 50 negative samples. In all, the G-DiaParaTrio multiplex PCR assay identified 38 G. intestinalis, 25 C. parvum/C. hominis and five E. histolytica leading to sensitivity/specificity of 92%/100%, 96%/100% and 100%/100% for G. intestinalis, C. parvum/C. hominis and E. histolytica, respectively. This new multiplex PCR assay offers fast and reliable results, similar to microscopy-driven diagnosis for the detection of these gastrointestinal protozoa, allowing its implementation in routine clinical practice. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  7. Using the Bristol Stool Scale and Parental Report of Stool Consistency as Part of the Rome III Criteria for Functional Constipation in Infants and Toddlers.

    Science.gov (United States)

    Koppen, Ilan J N; Velasco-Benitez, Carlos A; Benninga, Marc A; Di Lorenzo, Carlo; Saps, Miguel

    2016-10-01

    To evaluate among parents of infants and toddlers the agreement between parental report and the Bristol Stool Scale (BSS) in assessing stool consistency and the effect of both methods on determining the prevalence of functional constipation (FC) according to the Rome III criteria. Parents of children ≤48 months of age who were seen for a well-child visit completed a questionnaire about their child's bowel habits during the previous month. Cohen kappa coefficient (κ) was used to measure intrarater agreement between parental report of stool consistency ("hard," "normal," "soft/mucous/liquid") and the BSS (types 1-2, hard; types 3-5, normal; types 6-7, loose/liquid). The prevalence of FC was assessed based on the questionnaire according to the Rome III criteria, comparing both methods of stool consistency assessment. Parents of 1095 children (median age, 15 months; range, 1-48) were included. Only fair agreement existed between the 2 methods of stool consistency assessment (κ = 0.335; P < .001). According to the Rome III criteria, using parental report the prevalence of FC was 20.5% and using the BSS the prevalence was 20.9% (P = .87). The agreement between these 2 methods for assessing the prevalence of FC was excellent (κ = 0.95; P < .001). Only fair agreement exists between the BSS and parental report of stool consistency among parents of infants and toddlers. Different methods of stool consistency assessment did not result in a difference in the prevalence of FC. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. How to: Establish and run a stool bank.

    Science.gov (United States)

    Terveer, E M; van Beurden, Y H; Goorhuis, A; Seegers, J F M L; Bauer, M P; van Nood, E; Dijkgraaf, M G W; Mulder, C J J; Vandenbroucke-Grauls, C M J E; Verspaget, H W; Keller, J J; Kuijper, E J

    2017-12-01

    Since 2013, several stool banks have been developed following publications reporting on clinical success of 'faecal microbiota transplantation' (FMT) for recurrent Clostridium difficile infections (CDI). However, protocols for donor screening, faecal suspension preparation, and transfer of the faecal suspension differ between countries and institutions. Moreover, no European consensus exists regarding the legislative aspects of the faecal suspension product. Internationally standardized recommendations about the above mentioned aspects have not yet been established. In 2015, the Netherlands Donor Feces Bank (NDFB) was founded with the primary aim of providing a standardized product for the treatment of patients with recurrent CDI in the Netherlands. Standard operation procedures for donor recruitment, donor selection, donor screening, and production, storage, and distribution of frozen faecal suspensions for FMT were formulated. Our experience summarized in this review addresses current donor recruitment and screening, preparation of the faecal suspension, transfer of the faecal microbiota suspension, and the experiences and follow-up of the patients treated with donor faeces from the NDFB. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  9. Detection of Campylobacter in Stool and Determination of Significance by Culture, Enzyme Immunoassay, and PCR in Developing Countries

    Science.gov (United States)

    Platts-Mills, James A.; Liu, Jie; Gratz, Jean; Mduma, Esto; Amour, Caroline; Swai, Ndealilia; Taniuchi, Mami; Begum, Sharmin; Peñataro Yori, Pablo; Tilley, Drake H.; Lee, Gwenyth; Shen, Zeli; Whary, Mark T.; Fox, James G.; McGrath, Monica; Kosek, Margaret; Haque, Rashidul

    2014-01-01

    Campylobacter is a common bacterial enteropathogen that can be detected in stool by culture, enzyme immunoassay (EIA), or PCR. We compared culture for C. jejuni/C. coli, EIA (ProSpecT), and duplex PCR to distinguish Campylobacter jejuni/C. coli and non-jejuni/coli Campylobacter on 432 diarrheal and matched control stool samples from infants in a multisite longitudinal study of enteric infections in Tanzania, Bangladesh, and Peru. The sensitivity and specificity of culture were 8.5% and 97.6%, respectively, compared with the results of EIA and 8.7% and 98.0%, respectively, compared with the results of PCR for C. jejuni/C. coli. Most (71.6%) EIA-positive samples were positive by PCR for C. jejuni/C. coli, but 27.6% were positive for non-jejuni/coli Campylobacter species. Sequencing of 16S rRNA from 53 of these non-jejuni/coli Campylobacter samples showed that it most closely matched the 16S rRNA of C. hyointestinalis subsp. lawsonii (56%), C. troglodytis (33%), C. upsaliensis (7.7%), and C. jejuni/C. coli (2.6%). Campylobacter-negative stool spiked with each of the above-mentioned Campylobacter species revealed reactivity with EIA. PCR detection of Campylobacter species was strongly associated with diarrhea in Peru (odds ratio [OR] = 3.66, P < 0.001) but not in Tanzania (OR = 1.56, P = 0.24) or Bangladesh (OR = 1.13, P = 0.75). According to PCR, Campylobacter jejuni/C. coli infections represented less than half of all infections with Campylobacter species. In sum, in infants in developing country settings, the ProSpecT EIA and PCR for Campylobacter reveal extremely high rates of positivity. We propose the use of PCR because it retains high sensitivity, can ascertain burden, and can distinguish between Campylobacter infections at the species level. PMID:24452175

  10. Detection of Campylobacter in stool and determination of significance by culture, enzyme immunoassay, and PCR in developing countries.

    Science.gov (United States)

    Platts-Mills, James A; Liu, Jie; Gratz, Jean; Mduma, Esto; Amour, Caroline; Swai, Ndealilia; Taniuchi, Mami; Begum, Sharmin; Peñataro Yori, Pablo; Tilley, Drake H; Lee, Gwenyth; Shen, Zeli; Whary, Mark T; Fox, James G; McGrath, Monica; Kosek, Margaret; Haque, Rashidul; Houpt, Eric R

    2014-04-01

    Campylobacter is a common bacterial enteropathogen that can be detected in stool by culture, enzyme immunoassay (EIA), or PCR. We compared culture for C. jejuni/C. coli, EIA (ProSpecT), and duplex PCR to distinguish Campylobacter jejuni/C. coli and non-jejuni/coli Campylobacter on 432 diarrheal and matched control stool samples from infants in a multisite longitudinal study of enteric infections in Tanzania, Bangladesh, and Peru. The sensitivity and specificity of culture were 8.5% and 97.6%, respectively, compared with the results of EIA and 8.7% and 98.0%, respectively, compared with the results of PCR for C. jejuni/C. coli. Most (71.6%) EIA-positive samples were positive by PCR for C. jejuni/C. coli, but 27.6% were positive for non-jejuni/coli Campylobacter species. Sequencing of 16S rRNA from 53 of these non-jejuni/coli Campylobacter samples showed that it most closely matched the 16S rRNA of C. hyointestinalis subsp. lawsonii (56%), C. troglodytis (33%), C. upsaliensis (7.7%), and C. jejuni/C. coli (2.6%). Campylobacter-negative stool spiked with each of the above-mentioned Campylobacter species revealed reactivity with EIA. PCR detection of Campylobacter species was strongly associated with diarrhea in Peru (odds ratio [OR] = 3.66, P PCR, Campylobacter jejuni/C. coli infections represented less than half of all infections with Campylobacter species. In sum, in infants in developing country settings, the ProSpecT EIA and PCR for Campylobacter reveal extremely high rates of positivity. We propose the use of PCR because it retains high sensitivity, can ascertain burden, and can distinguish between Campylobacter infections at the species level.

  11. Stool screening of Syrian refugees and asylum seekers in Germany, 2013/2014: Identification of Sabin like polioviruses.

    Science.gov (United States)

    Böttcher, Sindy; Neubauer, Katrin; Baillot, Armin; Rieder, Gabriele; Adam, Maja; Diedrich, Sabine

    2015-10-01

    Germany is a partner of the Global Polio Eradication Initiative. Assurance of polio free status is based on enterovirus surveillance, which focuses on patients with signs of acute flaccid paralysis or aseptic meningitis/encephalitis, representing the key symptoms of poliovirus infection. In response to the wild poliovirus outbreak in Syria 2013 and high number of refugees coming from Syria to Germany, stool samples from 629 Syrian refugees/asylum seekers aged Syrian refugees and asylum seekers at that time. Copyright © 2015 Elsevier GmbH. All rights reserved.

  12. Stool frequency recording in severe acute malnutrition ('StoolSAM'); an agreement study comparing maternal recall versus direct observation using diapers.

    Science.gov (United States)

    Voskuijl, Wieger; Potani, Isabel; Bandsma, Robert; Baan, Anne; White, Sarah; Bourdon, Celine; Kerac, Marko

    2017-06-07

    Approximately 50% of the deaths of children under the age of 5 can be attributed to undernutrition, which also encompasses severe acute malnutrition (SAM). Diarrhoea is strongly associated with these deaths and is commonly diagnosed solely based on stool frequency and consistency obtained through maternal recall. This trial aims to determine whether this approach is equivalent to a 'directly observed method' in which a health care worker directly observed stool frequency using diapers in hospitalised children with complicated SAM. This study was conducted at 'Moyo' Nutritional Rehabilitation Unit, Queen Elizabeth Central Hospital, Malawi. Participants were children aged 5-59 months admitted with SAM. We compared 2 days of stool frequency data obtained with next-day maternal-recall versus a 'gold standard' in which a health care worker observed stool frequency every 2 h using diapers. After study completion, guardians were asked their preferred method and their level of education. We found poor agreement between maternal recall and the 'gold standard' of directly observed diapers. The sensitivity to detect diarrhoea based on maternal recall was poor, with only 75 and 56% of diarrhoea cases identified on days 1 and 2, respectively. However, the specificity was higher with more than 80% of children correctly classified as not having diarrhoea. On day 1, the mean stool frequency difference between the two methods was -0.17 (SD; 1.68) with limits of agreement (of stool frequency) of -3.55 and 3.20 and, similarly on day 2, the mean difference was -0.2 (SD; 1.59) with limits of agreement of -3.38 and 2.98. These limits extend beyond the pre-specified 'acceptable' limits of agreement (±1.5 stool per day) and indicate that the 2 methods are non-equivalent. The higher the stool frequency, the more discrepant the two methods were. Most primary care givers strongly preferred using diapers. This study shows lack of agreement between the assessment of stool frequency in SAM

  13. Evaluation of mixing downstream of tees in duct systems with respect to single point representative air sampling.

    Science.gov (United States)

    Kim, Taehong; O'Neal, Dennis L; Ortiz, Carlos

    2006-09-01

    Air duct systems in nuclear facilities must be monitored with continuous sampling in case of an accidental release of airborne radionuclides. The purpose of this work is to identify the air sampling locations where the velocity and contaminant concentrations fall below the 20% coefficient of variation required by the American National Standards Institute/Health Physics Society N13.1-1999. Experiments of velocity and tracer gas concentration were conducted on a generic "T" mixing system which included combinations of three sub ducts, one main duct, and air velocities from 0.5 to 2 m s (100 to 400 fpm). The experimental results suggest that turbulent mixing provides the accepted velocity coefficients of variation after 6 hydraulic diameters downstream of the T-junction. About 95% of the cases achieved coefficients of variation below 10% by 6 hydraulic diameters. However, above a velocity ratio (velocity in the sub duct/velocity in the main duct) of 2, velocity profiles were uniform in a shorter distance downstream of the T-junction as the velocity ratio went up. For the tracer gas concentration, the distance needed for the coefficients of variation to drop 20% decreased with increasing velocity ratio due to the sub duct airflow momentum. The results may apply to other duct systems with similar geometries and, ultimately, be a basis for selecting a proper sampling location under the requirements of single point representative sampling.

  14. Single-layer centrifugation separates spermatozoa from diploid cells in epididymal samples from gray wolves, Canis lupus (L.).

    Science.gov (United States)

    Muñoz-Fuentes, Violeta; Linde Forsberg, Catharina; Vilà, Carles; Morrell, Jane M

    2014-09-15

    Sperm samples may be used for assisted reproductive technologies (e.g., farmed or endangered species) or as a source of haploid DNA or sperm-specific RNA. When ejaculated spermatozoa are not available or are very difficult to obtain, as is the case for most wild endangered species, the epididymides of dead animals (e.g., animals that have been found dead, shot by hunters or poachers, or that that require euthanasia in zoological collections) can be used as a source of sperm. Such epididymal sperm samples are usually contaminated with cellular debris, erythrocytes, leukocytes, and sometimes also bacteria. These contaminants may be sources of reactive oxygen species that damage spermatozoa during freezing or contribute undesired genetic material from diploid cells. We used single-layer centrifugation through a colloid formulation, Androcoll-C, to successfully separate wolf epididymal spermatozoa from contaminating cells and cellular debris in epididymal samples harvested from carcasses. Such a procedure may potentially be applied to epididymal sperm samples from other species. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Mixed infection and clonal representativeness of a single sputum sample in tuberculosis patients from a penitentiary hospital in Georgia

    Directory of Open Access Journals (Sweden)

    Portaels Françoise

    2006-07-01

    Full Text Available Abstract Background Studies on recurrent tuberculosis (TB, TB molecular epidemiology and drug susceptibility testing rely on the analysis of one Mycobacterium tuberculosis isolate from a single sputum sample collected at different disease episodes. This scheme rests on the postulate that a culture of one sputum sample is homogeneous and representative of the total bacillary population in a patient. Methods We systematically analysed several pre-treatment isolates from each of 199 smear-positive male adult inmates admitted to a prison TB hospital by standard IS6110 DNA fingerprinting, followed by PCR typing based on multiple loci containing variable number of tandem repeats (VNTRs on a subset of isolates. Drug susceptibility testing (DST was performed on all isolates for isoniazid, rifampicin, streptomycin and ethambutol. Results We found mixed infection in 26 (13.1% cases. In contrast, analysis of a single pre-treatment isolate per patient would have led to missed mixed infections in all or 14 of these 26 cases by using only standard DNA fingerprinting or the PCR multilocus-based method, respectively. Differences in DST among isolates from the same patient were observed in 10 cases, of which 6 were from patients with mixed infection. Conclusion These results suggest that the actual heterogeneity of the bacillary population in patients, especially in high TB incidence settings, may be frequently underestimated using current analytical schemes. These findings have therefore important implications for correct interpretation and evaluation of molecular epidemiology data and in treatment evaluations.

  16. Consistency of direct microscopic examination and ELISA in detection of Giardia in stool specimen among children

    Directory of Open Access Journals (Sweden)

    Zohreh Torabi

    2014-09-01

    Full Text Available Objective: To investigate the consistency of direct microscopic examination and ELISA for determination of Giadia in stool specimen. Method: Study population consisted of children with any clinical symptoms of Giardia infestation since last two weeks. Fresh stool specimen was collected from each child. The stools specimens were assessed by two methods of direct microscopic examination and ELISA.The degree of agreement between direct stool exam and ELISA was calculated by Cohen's kappa coefficient. Results: In this study, 124 children with age range 2-12 years were investigated. A total of 64 (61.7% and 79 (65.7% of children had Giardia by direct stool exam and ELISA test respectively. There was association between frequency of constipation and Giardia infection (P=0.036. The Cohen's kappa coefficient calculated for degree of agreement between direct stool exam and ELISA showed κ=0.756 (P<0.001. Conclusions: The frequency of Giardia infection in symptomatic children was high and there was high agreement rate between ELISA and direct stool smear.

  17. Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool

    Directory of Open Access Journals (Sweden)

    Zoheiry Mona K

    2011-09-01

    Full Text Available Abstract Background This research was carried out to develop a reliable monoclonal antibody (MoAb-based sandwich enzyme linked immunosorbent assay (ELISA for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. Methods From a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags, a pair (12B/11D/3F and 10A/9D/10G was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA. Results The two MoAbs were of the IgG1 and IgG2a subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p Conclusions These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.

  18. Comparison of three stool antigen assays with the 13C- urea breath test for the primary diagnosis of Helicobacter pylori infection and monitoring treatment outcome.

    LENUS (Irish Health Repository)

    Hooton, Carmel

    2012-02-03

    BACKGROUND: The urea breath test (UBT) is the gold-standard non-invasive test for the detection of Helicobacter pylori infection, however, the lack of availability of the UBT due to the high cost of the test, and in particular the need for expensive analytical instrumentation, limits the usefulness of this method. Stool antigen assays may offer an alternative non-invasive method for the diagnosis of infection. OBJECTIVE: To compare the accuracy of three stool antigen assays (HpSA, IDEIA HpStAR, and ImmunoCard STAT) against the UBT for the primary diagnosis of H. pylori infection and for monitoring treatment outcome. METHODS: A total of 102 patients attending two gastroenterology day-case clinics for the investigation of dyspepsia were included. Each patient provided breath and stool samples for analysis. Patients who tested positive for H. pylori by the validated UBT were prescribed triple therapy and invited to return for repeat breath and stool sample analysis 6 weeks post-treatment. RESULTS: Of the 102 patients tested, 48 were diagnosed with H. pylori infection by the UBT. The HpSA assay interpreted 38 of these as positive (79% sensitive). Of the 54 UBT-negative patients the HpSA assay interpreted all 54 as negative (100% specific). The IDEIA HpStAR assay correctly identified 44 patients as positive (92% sensitive) and 50 as negative (92.5% specific). The ImmunoCard STAT assay interpreted 38 patients as positive (79% sensitive) and 52 as negative (96.3% specific). CONCLUSION: The findings indicate that the IDEIA HpStAR stool antigen kit is the most accurate assay of the three assays evaluated, and possibly represents a viable alternative to the UBT for the primary diagnosis of H. pylori infection and for monitoring treatment outcome.

  19. Relating Stool Microbial Metabolite Levels, Inflammatory Markers and Dietary Behaviors to Screening Colonoscopy Findings in a Racially/Ethnically Diverse Patient Population

    Directory of Open Access Journals (Sweden)

    Kristina M. Bridges

    2018-02-01

    Full Text Available Colorectal cancer (CRC is the third leading cause of cancer death for both men and women in the United States, yet it is treatable and preventable. African Americans have higher incidence of CRC than other racial/ethnic groups, however, it is unclear whether this disparity is primarily due to environmental or biological factors. Short chain fatty acids (SCFAs are metabolites produced by bacteria in the colon and are known to be inversely related to CRC progression. The aim of this study is to investigate how stool SCFA levels, markers of inflammation in stool and dietary intake relate to colonoscopy findings in a diverse patient population. Stool samples from forty-eight participants were analyzed for SCFA levels and inflammatory markers (lysozyme, secretory IgA, lactoferrin. Additionally, participants completed the National Cancer Institute’s Diet History Questionnaire II (DHQ II to report dietary intake over the past year. Subsequently, the majority of participants underwent screening colonoscopy. Our results showed that African Americans had higher total levels of SCFAs in stool than other racial/ethnic groups, significantly lower intake of non-starchy vegetables and similar inflammatory marker expression and colonoscopy outcomes, compared to others. This work is an initial exploration into the biological and clinical factors that may ultimately inform personalized screening approaches and clinical decision-making to improve colorectal cancer disparities for African Americans.

  20. Longitudinal studies of neutralizing antibody responses to rotavirus in stools and sera of children following severe rotavirus gastroenteritis.

    Science.gov (United States)

    Coulson, B S

    1998-11-01

    Rotavirus-neutralizing antibody responses in sera and stools of children hospitalized with rotavirus gastroenteritis and then monitored longitudinally were optimally detected by using local rotavirus strains. Stool responses were highest on days 5 to 8 after the onset of diarrhea. Longitudinal monitoring suggested that serum neutralizing antibody responses were a more useful measure of severely symptomatic rotavirus infection than stool responses but that stool antibody responses may be a useful measure of rotavirus immunity.

  1. Longitudinal Studies of Neutralizing Antibody Responses to Rotavirus in Stools and Sera of Children following Severe Rotavirus Gastroenteritis

    OpenAIRE

    Coulson, Barbara S.

    1998-01-01

    Rotavirus-neutralizing antibody responses in sera and stools of children hospitalized with rotavirus gastroenteritis and then monitored longitudinally were optimally detected by using local rotavirus strains. Stool responses were highest on days 5 to 8 after the onset of diarrhea. Longitudinal monitoring suggested that serum neutralizing antibody responses were a more useful measure of severely symptomatic rotavirus infection than stool responses but that stool antibody responses may be a use...

  2. Development of the Brussels Infant and Toddler Stool Scale ('BITSS'): protocol of the study.

    Science.gov (United States)

    Vandenplas, Yvan; Szajewska, Hania; Benninga, Marc; Di Lorenzo, Carlo; Dupont, Christophe; Faure, Christophe; Miqdadi, Mohamed; Osatakul, Seksit; Ribes-Konickx, Carmen; Saps, Miguel; Shamir, Raanan; Staiano, Annamaria

    2017-03-29

    The Bristol Stool Form Scale (BSS) which consists of 7 photographs of different stool forms allows assessment of stool consistency (scale 1 for hard lumps to scale 7 for watery stools), in an objective manner in adults. The BSS is also sometimes used to characterise the stools of infants and young children. Despite its use, there is general agreement among paediatric gastroenterologists that the BSS is not adequate to be used in infants and young children who wear diapers; thus, a new scale specifically designed for this population is needed. Our aim is to develop a paediatric stool scale, the Brussels Infant and Toddler Stool Scale ('BITSS'), and to evaluate the interobserver agreement of stool assessment with the BITSS between the patient's parent and healthcare providers (physicians and nurses). This study has two phases. In the first phase, 11 key-opinion leaders in the field of paediatric gastroenterology representing different areas of the world selected seven coloured photographs of infants and/or young children wearing diapers to match the original descriptors of the BSS. The selected photographs were used to create a new scale in which the drawings of stools of the BSS were replaced by infant/toddlers stool photographs. In phase II, we aim at demonstrating that parents, nurses and primary healthcare physicians interpret the stool-pictures of the BITSS with a high degree of consensus and that the agreement is independent of whether it is a parent or a healthcare provider. Interobserver variability of stool assessment with the BITSS between the patient's parent and healthcare providers will be assessed. The study will be approved by the Ethics Committee of the participating centres. The findings of this study will be submitted to a peer-reviewed journal. Abstracts will be submitted to national and international conferences. NCT02913950. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go

  3. Development of a novel method for simultaneous concentration of viruses and protozoa from a single water sample.

    Science.gov (United States)

    Haramoto, Eiji; Katayama, Hiroyuki; Asami, Mari; Akiba, Michihiro

    2012-06-01

    A novel method, electronegative membrane-vortex (EMV) method, was developed for simultaneous concentration of viruses and protozoa from a single water sample. Viruses and protozoa in a water sample were mixed with a cation solution and adsorbed on an electronegative membrane. Concentrated virus and protozoa samples were obtained as supernatant and pellet fractions, respectively, by vigorous vortex mixing of the membrane and centrifugation of the eluted material. The highest recovery efficiencies of model microbes from river water and tap water by this EMV method were obtained using a mixed cellulose ester membrane with a pore size of 0.45 μm (Millipore) as the electronegative membrane and MgCl(2) as the cation solution. The recovery was 27.7-86.5% for poliovirus, 25.7-68.3% for coliphage Qβ, 28.0-60.0% for Cryptosporidium oocysts, and 35.0-53.0% for Giardia cysts. The EMV method detected successfully indigenous viruses and protozoa in wastewater and river water samples from the Kofu basin, Japan, showing an overall positive rate of 100% (43/43) for human adenovirus, 79% (34/43) for norovirus GI, 65% (28/43) for norovirus GII, 23% (10/43) for Cryptosporidium oocysts, and 60% (26/43) for Giardia cysts. By direct DNA sequencing, a total of four genotypes (AI, AII, B, and G) of Giardia intestinalis were identified in the water samples, indicating that the river water was contaminated with feces from various mammals, including humans. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Optimization of plasma sampling depth and aerosol gas flow rates for single particle inductively coupled plasma mass spectrometry analysis.

    Science.gov (United States)

    Kálomista, Ildikó; Kéri, Albert; Galbács, Gábor

    2017-09-01

    We performed experiments to assess the separate and also the combined effect of the sampling depth and the aerosol gas flow rates on the signal formation in single particle inductively coupled plasma mass spectrometry (spICP-MS) measurements by using dispersions containing Ag and Au NPs. It was found that the NP signal can significantly be improved by the optimization of the sampling depth. With respect to the "robust" setting, a signal improvement of nearly 100% could be achieved, which translates into a 25-30% improvement in size detection limits. It was also found that the shape of the spICP-MS signal histograms also change with the change of the plasma sampling depth. It was demonstrated that nanoparticle peak separation can also be significantly enhanced by using sampling depth optimization. The effect of the aerosol dilution gas flow, now standard in most ICP-MS instruments, on the spICP-MS signal formation was also studied for the first time in the literature, as this flow was hoped to make spICP-MS measurements more practical and faster via the on-line dilution of the aerosol generated from nano-dispersions. Our experimental results revealed that the dilution gas flow can only be used for a moderate aerosol dilution in spICP-MS measurements, if the gas flow going to the pneumatic nebulizer is proportionally lowered at the same time. This however was found to cause a significant worsening in the operation of the sample introduction system, which gives rise to a strong NP signal loss. Thus it was concluded that the use of the aerosol dilution gas flow, in its present form, can not be suggested for spICP-MS analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample.

    Science.gov (United States)

    Salem, Mohamed; Bernach, Michal; Bajdzienko, Krzysztof; Giavalisco, Patrick

    2017-06-01

    Understanding of complex biological systems requires the measurement, analysis and integration of multiple compound classes of the living cell, usually determined by transcriptomic, proteomic, metabolomics and lipidomic measurements. In this protocol, we introduce a simple method for the reproducible extraction of metabolites, lipids and proteins from biological tissues using a single aliquot per sample. The extraction method is based on a methyl tert-butyl ether: methanol: water system for liquid: liquid partitioning of hydrophobic and polar metabolites into two immiscible phases along with the precipitation of proteins and other macromolecules as a solid pellet. This method, therefore, provides three different fractions of specific molecular composition, which are fully compatible with common high throughput 'omics' technologies such as liquid chromatography (LC) or gas chromatography (GC) coupled to mass spectrometers. Even though the method was initially developed for the analysis of different plant tissue samples, it has proved to be fully compatible for the extraction and analysis of biological samples from systems as diverse as algae, insects, and mammalian tissues and cell cultures.

  6. Could stool collection devices help increase uptake in bowel cancer screening programmes?

    Science.gov (United States)

    Morling, J R; Barke, A N; Chapman, C J; Logan, R F

    2018-01-01

    Objective To understand the usage and acceptability of a faecal collection device amongst participants in the National Health Service Bowel Cancer Screening Programme, with the aim of influencing future uptake. Setting Participants completing faecal occult blood test retests as part of the routine Bowel Cancer Screening Programme in Eastern England. Methods A faecal collection device and questionnaire were sent to all potential retest participants during a one-month period to collect information on prior stool collection methods and ease of use and usefulness of the enclosed faecal collection device. Results Out of 1087 participants invited, 679 (62.5%) returned their questionnaire. Of these, 429 (63.2%) trialled the faecal collection device at least once, 163 (38.4%) found the device made collecting their sample easier than previously, 189 (44.6%) found it made collection more difficult and 72 (17.0%) said it made no difference. Similar numbers reported finding that the faecal collection device made collecting the sample more pleasant (130, 31.5%), less pleasant (103, 25.0%) and no different (179, 43.4%) compared with previous collection without a faecal collection device. Conclusion Although a small proportion of participants found the faecal collection device helpful, a considerable majority did not or did not use it at all. Offering faecal collection devices is unlikely to produce a substantial increase in bowel cancer screening uptake.

  7. Integrative analysis of single nucleotide polymorphisms and gene expression efficiently distinguishes samples from closely related ethnic populations

    Directory of Open Access Journals (Sweden)

    Yang Hsin-Chou

    2012-07-01

    Full Text Available Abstract Background Ancestry informative markers (AIMs are a type of genetic marker that is informative for tracing the ancestral ethnicity of individuals. Application of AIMs has gained substantial attention in population genetics, forensic sciences, and medical genetics. Single nucleotide polymorphisms (SNPs, the materials of AIMs, are useful for classifying individuals from distinct continental origins but cannot discriminate individuals with subtle genetic differences from closely related ancestral lineages. Proof-of-principle studies have shown that gene expression (GE also is a heritable human variation that exhibits differential intensity distributions among ethnic groups. GE supplies ethnic information supplemental to SNPs; this motivated us to integrate SNP and GE markers to construct AIM panels with a reduced number of required markers and provide high accuracy in ancestry inference. Few studies in the literature have considered GE in this aspect, and none have integrated SNP and GE markers to aid classification of samples from closely related ethnic populations. Results We integrated a forward variable selection procedure into flexible discriminant analysis to identify key SNP and/or GE markers with the highest cross-validation prediction accuracy. By analyzing genome-wide SNP and/or GE markers in 210 independent samples from four ethnic groups in the HapMap II Project, we found that average testing accuracies for a majority of classification analyses were quite high, except for SNP-only analyses that were performed to discern study samples containing individuals from two close Asian populations. The average testing accuracies ranged from 0.53 to 0.79 for SNP-only analyses and increased to around 0.90 when GE markers were integrated together with SNP markers for the classification of samples from closely related Asian populations. Compared to GE-only analyses, integrative analyses of SNP and GE markers showed comparable testing

  8. Evaluation of Patients with an Apparent False Positive Stool DNA Test: The Role of Repeat Stool DNA Testing.

    Science.gov (United States)

    Cooper, Gregory S; Markowitz, Sanford D; Chen, Zhengyi; Tuck, Missy; Willis, Joseph E; Berger, Barry M; Brenner, Dean E; Li, Li

    2018-03-07

    There is uncertainty as to the appropriate follow-up of patients who test positive on multimarker stool DNA (sDNA) testing and have a colonoscopy without neoplasia. To determine the prevalence of missed colonic or occult upper gastrointestinal neoplasia in patients with an apparent false positive sDNA. We prospectively identified 30 patients who tested positive with a commercially available sDNA followed by colonoscopy without neoplastic lesions. Patients were invited to undergo repeat sDNA at 11-29 months after the initial test followed by repeat colonoscopy and upper endoscopy. We determined the presence of neoplastic lesions on repeat evaluation stratified by results of repeat sDNA. Twelve patients were restudied. Seven patients had a negative second sDNA test and a normal second colonoscopy and upper endoscopy. In contrast, 5 of 12 subjects had a persistently positive second sDNA test, and 3 had positive findings, including a 3-cm sessile transverse colon adenoma with high-grade dysplasia, a 2-cm right colon sessile serrated adenoma with dysplasia, and a nonadvanced colon adenoma (p = 0.045). These corresponded to a positive predictive value of 0.60 (95% CI 0.17-1.00) and a negative predictive value of 1.00 (95% CI 1.00-1.00) for the second sDNA test. In addition, the medical records of all 30 subjects with apparent false positive testing were reviewed and no documented cases of malignant tumors were recorded. Repeat positive sDNA testing may identify a subset of patients with missed or occult colorectal neoplasia after negative colonoscopy for an initially positive sDNA. High-quality colonoscopy with careful attention to the right colon in patients with positive sDNA is critically important and may avoid false negative colonoscopy.

  9. Classifying the Progression of Ductal Carcinoma from Single-Cell Sampled Data via Integer Linear Programming: A Case Study.

    Science.gov (United States)

    Catanzaro, Daniele; Shackney, Stanley E; Schaffer, Alejandro A; Schwartz, Russell

    2016-01-01

    Ductal Carcinoma In Situ (DCIS) is a precursor lesion of Invasive Ductal Carcinoma (IDC) of the breast. Investigating its temporal progression could provide fundamental new insights for the development of better diagnostic tools to predict which cases of DCIS will progress to IDC. We investigate the problem of reconstructing a plausible progression from single-cell sampled data of an individual with synchronous DCIS and IDC. Specifically, by using a number of assumptions derived from the observation of cellular atypia occurring in IDC, we design a possible predictive model using integer linear programming (ILP). Computational experiments carried out on a preexisting data set of 13 patients with simultaneous DCIS and IDC show that the corresponding predicted progression models are classifiable into categories having specific evolutionary characteristics. The approach provides new insights into mechanisms of clonal progression in breast cancers and helps illustrate the power of the ILP approach for similar problems in reconstructing tumor evolution scenarios under complex sets of constraints.

  10. Helminth prevalence among adults in rural Kenya: a stool survey for soil-transmitted helminths and schistosomiasis in Nyanza province.

    Science.gov (United States)

    Andereck, Jonathan W; Kipp, Aaron M; Ondiek, Michael; Vermund, Sten H

    2014-12-01

    Soil-transmitted helminth (STH) prevalence in children is high in rural southwestern Kenya, but adult prevalence data are scarce. A 2010 study of a village in Nyanza province found a pediatric STH prevalence of 44% using a direct stool-smear method. Adult STH prevalence and associated predictors was measured in the same village. Adults (≥18 years) presenting at the out-patient department of the small hospital or community outreach events completed a short questionnaire and provided stool samples. Light microscopy for ova and larvae was conducted using a stool concentration technique to improve sensitivity. Multivariable regression models were used to identify predictors of STH prevalence. Among 344 adults, STH prevalence was 15.7% (54/344). Hookworm was most common (13.1%; 45/344), followed by Ascaris lumbricoides (6.1%; 21/344) and Trichuris trichiura (0.6%; 2/344). Twelve participants (3.5%; 12/344) had multiple STHs and three (0.9%; 3/344) had Schistosoma mansoni. Female sex, older age and lower education level were significant STH predictors. Adult STH prevalence was lower than previous studies of children from the same village. Adults with the identified risk factors had a prevalence of ≥20%, which may warrant periodic, targeted deworming of adults with these risk factors given the low cost and low toxicity of anthelmintic drugs. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Algorithm and program for precise determination of unit-cell parameters of single crystal taking into account the sample eccentricity

    Science.gov (United States)

    Dudka, A. P.; Smirnova, E. S.; Verin, I. A.; Bolotina, N. B.

    2017-07-01

    A technique has been developed to refine the unit-cell parameters of single crystals with minimization of the influence of instrumental errors on the result. The corresponding computational procedure HuberUB is added to the software package of Huber-5042 diffractometer with a point detector and closedcycle helium cryostat Displex DE-202. The parameters of unit cell, its orientation, the goniometer zero angles, the sample eccentricity, the distances in the goniometer, and the radiation wavelength were refined by the nonlinear least-squares method, which allows imposition of constraints on the unit-cell parameters, depending on the crystal symmetry. The technique is approved on a LuB12 single crystal. The unit-cell parameters are determined in a temperature range of 20-295 K, with an absolute error not larger than 0.0004 Å (the relative error is of 5 × 10-5). The estimates of the unit-cell parameters obtained by the proposed method are evidenced to be unbiased. Some specific features of the behavior of parameters in the ranges of 120-140 and 20-50 K are revealed, which correlate with the anomalies of the physical properties of the crystal.

  12. Antimicrobial resistance of bacterial enteropathogens isolated from stools in Madagascar.

    Science.gov (United States)

    Randrianirina, Frederique; Ratsima, Elisoa Hariniana; Ramparany, Lova; Randremanana, Rindra; Rakotonirina, Hanitra Clara; Andriamanantena, Tahiry; Rakotomanana, Fanjasoa; Rajatonirina, Soatiana; Richard, Vincent; Talarmin, Antoine

    2014-02-25

    Diarrheal diseases are a major public health problem in developing countries, and are one of the main causes of hospital admissions in Madagascar. The Pasteur Institute of Madagascar undertook a study to determine the prevalence and the pathogenicity of bacterial, viral and protozoal enteropathogens in diarrheal and non-diarrheal stools of children aged less than 5 years in Madagascar. We present here the results of the analysis of antimicrobial susceptibility of the bacteria isolated during this study. The study was conducted in the community setting in 14 districts of Madagascar from October 2008 to May 2009. Conventional methods and PCR were used to identify the bacteria; antimicrobial susceptibility was determined using an agar diffusion method for enterobacteriaceae and MICs were measured by an agar dilution method for Campylobacter sp. In addition to the strains isolated during this study, Salmonella sp and Shigella sp isolated at the Pasteur Institute of Madagascar from 2005 to 2009 were included in the analysis to increase the power of the study. Twenty-nine strains of Salmonella sp, 35 strains of Shigella sp, 195 strains of diarrheagenic E. coli, 203 strains of C. jejuni and 71 strains of C. coli isolated in the community setting were tested for antibiotic resistance. Fifty-five strains of Salmonella sp and 129 strains of Shigella sp isolated from patients referred to the Pasteur Institute of Madagascar were also included in the study. Many E. coli and Shigella isolates (around 80%) but fewer Salmonella isolates were resistant to ampicillin and trimethoprim/sulfamethoxazole. A small proportion of strains of each species were resistant to ciprofloxacin and only 3% of E. coli strains presented a resistance to third generation cephalosporins due to the production of extended-spectrum beta-lactamases. The resistance of Campylobacter sp to ampicillin was the most prevalent, whereas less than 5% of isolates were resistant to each of the other antibiotics. The

  13. Feeding, stooling and sleeping patterns in infants with colic - a randomized controlled trial of minimal acupuncture

    Directory of Open Access Journals (Sweden)

    Hallström Inger

    2011-10-01

    Full Text Available Abstract Background The aim was to describe the feeding- and stooling patterns of infants with colic and evaluate the influence of minimal acupuncture. Methods A prospective, randomized, controlled, blind clinical study was conducted at a private acupuncture clinic in Sweden. 90 otherwise healthy 2-8 weeks old infants, born after gestational week 36, fulfilling the criteria for infantile colic and not medicated with dicyclomine, were included. 81 infants went through a structured program consisting of six visits to the clinic, twice weekly. Infants randomized to receive acupuncture were given minimal, standardized acupuncture for two seconds in LI4. Frequency and size of stooling, as well as duration of, and intervals between, feeding sessions were reported by parents in a diary. Parental assessment of sleep and comments on stooling and side effects were collected in a questionnaire. Results At baseline when the mean age was five weeks, infants in both groups were fed a median of eight times/day, 148 min/day, with considerable variations. No differences were found between groups in the frequency and duration of feeding during the intervention weeks. Furthermore there were no significant differences between the groups regarding the frequency of stooling, neither at baseline, at which point the infants of both groups had bowel movements 4.2 times/day, nor during the intervention weeks. There was an expected decrease in frequency of stooling in both groups, reaching 2.1 (p = 0,001 in the acupuncture group and 3.1 (p Conclusions Infants with colic in the present study had a higher frequency of stooling than reported internationally in healthy infants. Minimal acupuncture had no major effect on feeding, stooling and sleep, although a minor effect of minimal acupuncture on stooling and sleep cannot be ruled out. Trial registration ClinicalTrials.govID NCT00860301

  14. Aeromonas Isolates from Human Diarrheic Stool and Groundwater Compared by Pulsed-Field Gel Electrophoresis

    OpenAIRE

    Borchardt, Mark A.; Stemper, Mary E.; Standridge, Jon H.

    2003-01-01

    Gastrointestinal infections of Aeromonas species are generally considered waterborne; for this reason, Aeromonas hydrophila has been placed on the United States Environmental Protection Agency Contaminant Candidate List of emerging pathogens in drinking water. In this study, we compared pulsed-field gel electrophoresis patterns of Aeromonas isolates from stool specimens of patients with diarrhea with Aeromonas isolates from patients? drinking water. Among 2,565 diarrheic stool specimens submi...

  15. pH(stat) vs. single extraction tests to evaluate heavy metals and arsenic leachability in environmental samples.

    Science.gov (United States)

    Rigol, A; Mateu, J; González-Núñez, R; Rauret, G; Vidal, M

    2009-01-19

    Here we compared the pH(stat) test, which examines the leachability of major elements (Ca, Mg, Al, Fe, and Mn), dissolved organic carbon, and trace elements (Cd, Zn, Cu, Pb, and As) in a wide pH range, with single extraction tests based on the use of mild extractants (calcium chloride, acetic acid or EDTA). For this purpose, we examined samples from a variety of environmental conditions (sludges, mineral soils, organic soils, and soils with particulate and/or soluble contamination). Extraction yields obtained with CaCl(2) (0.01 mol L(-1)) and CH(3)COOH (0.43 mol L(-1)) correlated well with those from the pH(stat) at the same pH (r=0.98 and 0.95, respectively), while the use of EDTA (0.05 mol L(-1)) led to systematically higher extraction yields than those quantified with the pH(stat) at the same pH. However, the pH(stat) test had three distinct advantages: (1) it revealed the relationship between the solubility of the main soil phases and pH; (2) it showed the variation in pollutant leachability due to changes in pH; and (3) it better predicted the maximum contaminant availability. Thus we propose that the pH(stat) is the best laboratory tests to evaluate the contaminant leachability over a wide range of sample types (soil, sludge, and sediment).

  16. Microbiome Analysis of Stool Samples from African Americans with Colon Polyps

    NARCIS (Netherlands)

    Brim, H.; Yooseph, S.; Zoetendal, E.G.; Lee, E.; Torralbo, M.; Laiyemo, A.O.; Shokrani, B.; Nelson, K.; Ashktorab, H.

    2013-01-01

    Background: Colonic polyps are common tumors occurring in similar to 50% of Western populations with similar to 10% risk of malignant progression. Dietary agents have been considered the primary environmental exposure to promote colorectal cancer (CRC) development. However, the colonic mucosa is

  17. Remediation of intramacrophageal Shigella dysenteriae type 1 by probiotic lactobacilli isolated from human infants' stool samples

    Directory of Open Access Journals (Sweden)

    Radhika Trikha

    2017-01-01

    Interpretation & conclusions: Probiotic cocktail of L. pentosus, L. paraplantarum and L. rhamnosus showed ex vivo killing of S. dysenteriae residing inside the rat macrophages significantly. This cocktail has the potential to be used as a natural alternative for treating S. dysenteriae infection, especially in infants, however, further studies need to be done to confirm these finding in vivo.

  18. Stool frequency recording in severe acute malnutrition ('StoolSAM'); an agreement study comparing maternal recall versus direct observation using diapers

    NARCIS (Netherlands)

    Voskuijl, Wieger; Potani, Isabel; Bandsma, Robert; Baan, Anne; White, Sarah; Bourdon, Celine; Kerac, Marko

    2017-01-01

    Background: Approximately 50% of the deaths of children under the age of 5 can be attributed to undernutrition, which also encompasses severe acute malnutrition (SAM). Diarrhoea is strongly associated with these deaths and is commonly diagnosed solely based on stool frequency and consistency

  19. Single Particle-Inductively Coupled Plasma Mass Spectroscopy Analysis of Metallic Nanoparticles in Environmental Samples with Large Dissolved Analyte Fractions.

    Science.gov (United States)

    Schwertfeger, D M; Velicogna, Jessica R; Jesmer, Alexander H; Scroggins, Richard P; Princz, Juliska I

    2016-10-18

    There is an increasing interest to use single particle-inductively coupled plasma mass spectroscopy (SP-ICPMS) to help quantify exposure to engineered nanoparticles, and their transformation products, released into the environment. Hindering the use of this analytical technique for environmental samples is the presence of high levels of dissolved analyte which impedes resolution of the particle signal from the dissolved. While sample dilution is often necessary to achieve the low analyte concentrations necessary for SP-ICPMS analysis, and to reduce the occurrence of matrix effects on the analyte signal, it is used here to also reduce the dissolved signal relative to the particulate, while maintaining a matrix chemistry that promotes particle stability. We propose a simple, systematic dilution series approach where by the first dilution is used to quantify the dissolved analyte, the second is used to optimize the particle signal, and the third is used as an analytical quality control. Using simple suspensions of well characterized Au and Ag nanoparticles spiked with the dissolved analyte form, as well as suspensions of complex environmental media (i.e., extracts from soils previously contaminated with engineered silver nanoparticles), we show how this dilution series technique improves resolution of the particle signal which in turn improves the accuracy of particle counts, quantification of particulate mass and determination of particle size. The technique proposed here is meant to offer a systematic and reproducible approach to the SP-ICPMS analysis of environmental samples and improve the quality and consistency of data generated from this relatively new analytical tool.

  20. Pharmacokinetics of tulathromycin in plasma and milk samples after a single subcutaneous injection in lactating goats (Capra hircus).

    Science.gov (United States)

    Grismer, B; Rowe, J D; Carlson, J; Wetzlich, S E; Tell, L A

    2014-04-01

    Eight adult female dairy goats received one subcutaneous administration of tulathromycin at a dosage of 2.5 mg/kg body weight. Blood and milk samples were assayed for tulathromycin and the common fragment of tulathromycin, respectively, using liquid chromatography/mass spectrometry. Pharmacokinetic disposition of tulathromycin was analyzed by a noncompartmental approach. Mean plasma pharmacokinetic parameters (±SD) following single-dose administration of tulathromycin were as follows: C(max) (121.54 ± 19.01 ng/mL); T(max) (12 ± 12-24 h); area under the curve AUC(0→∞) (8324.54 ± 1706.56 ng·h/mL); terminal-phase rate constant λz (0.01 ± 0.002 h⁻¹); and terminal-phase rate constant half-life t1/2λz (67.20 h; harmonic). Mean milk pharmacokinetic parameters (±SD) following 45 days of sampling were as follows: Cmax (1594 ± 379.23 ng/mL); Tmax (12 ± 12-36 h); AUC(0→∞) (72,250.51 ± 18,909.57 ng·h/mL); λz (0.005 ± 0.001 h⁻¹); and t(1/2λz) (155.28 h; harmonic). All goats had injection-site reactions that diminished in size over time. The conclusions from this study were that tulathromycin residues are detectable in milk samples from adult goats for at least 45 days following subcutaneous administration, this therapeutic option should be reserved for cases where other treatment options have failed, and goat milk should be withheld from the human food chain for at least 45 days following tulathromycin administration. © 2013 John Wiley & Sons Ltd.

  1. Quantification and size characterisation of silver nanoparticles in environmental aqueous samples and consumer products by single particle-ICPMS.

    Science.gov (United States)

    Aznar, Ramón; Barahona, Francisco; Geiss, Otmar; Ponti, Jessica; José Luis, Tadeo; Barrero-Moreno, Josefa

    2017-12-01

    Single particle-inductively coupled plasma mass spectrometry (SP-ICPMS) is a promising technique able to generate the number based-particle size distribution (PSD) of nanoparticles (NPs) in aqueous suspensions. However, SP-ICPMS analysis is not consolidated as routine-technique yet and is not typically applied to real test samples with unknown composition. This work presents a methodology to detect, quantify and characterise the number-based PSD of Ag-NPs in different environmental aqueous samples (drinking and lake waters), aqueous samples derived from migration tests and consumer products using SP-ICPMS. The procedure is built from a pragmatic view and involves the analysis of serial dilutions of the original sample until no variation in the measured size values is observed while keeping particle counts proportional to the dilution applied. After evaluation of the analytical figures of merit, the SP-ICPMS method exhibited excellent linearity (r 2 >0.999) in the range (1-25) × 10 4 particlesmL -1 for 30, 50 and 80nm nominal size Ag-NPs standards. The precision in terms of repeatability was studied according to the RSDs of the measured size and particle number concentration values and a t-test (p = 95%) at the two intermediate concentration levels was applied to determine the bias of SP-ICPMS size values compared to reference values. The method showed good repeatability and an overall acceptable bias in the studied concentration range. The experimental minimum detectable size for Ag-NPs ranged between 12 and 15nm. Additionally, results derived from direct SP-ICPMS analysis were compared to the results conducted for fractions collected by asymmetric flow-field flow fractionation and supernatant fractions after centrifugal filtration. The method has been successfully applied to determine the presence of Ag-NPs in: lake water; tap water; tap water filtered by a filter jar; seven different liquid silver-based consumer products; and migration solutions (pure water and

  2. Genotyping of human parvovirus B19 in clinical samples from Brazil and Paraguay using heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing

    Directory of Open Access Journals (Sweden)

    Marcos César Lima de Mendonça

    2011-06-01

    Full Text Available Heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing were utilised to genotype human parvovirus B19 samples from Brazil and Paraguay. Ninety-seven serum samples were collected from individuals presenting with abortion or erythema infectiosum, arthropathies, severe anaemia and transient aplastic crisis; two additional skin samples were collected by biopsy. After the procedure, all clinical samples were classified as genotype 1.

  3. Cryptosporidium infection in Bedouin infants assessed by prospective evaluation of anticryptosporidial antibodies and stool examination.

    Science.gov (United States)

    Robin, G; Fraser, D; Orr, N; Sela, T; Slepon, R; Ambar, R; Dagan, R; Le Blancq, S; Deckelbaum, R J; Cohen, D

    2001-01-15

    An enzyme-linked immunosorbent assay system using oocyst lysate as antigen was used to detect serum- specific antibody responses to Cryptosporidium parvum between 1989 and 1994 in consecutive sera obtained at birth, and at the age of 6, 12, and 23 months, from 52 infants living in a Bedouin town located in the south of Israel. The serologic tests revealed high levels of immunoglobulin G anti-Cryptosporidium at birth that dropped significantly by the age of 6 months and then rose continuously to a geometric mean titer of 481 at age 23 months. The serum immunoglobulin M Cryptosporidium antibodies rose continuously from nearly undetectable levels at birth to a geometric mean titer of 471 (157-fold increase) at age 23 months. All the subjects already showed at 6 months a significant rise in immunoglobulin M. A significant rise in immunoglobulin A titers was detected in 48% and 91% of subjects at 6 and 23 months, respectively. By monthly surveillance, microscopy using the modified Ziehl-Neelsen method and confirmed by indirect immunofluorescence assay detected Cryptosporidium antigens in only 11% at age 6 months and 48% at age 23 months. The extent of exposure to Cryptosporidium immediately after birth as detected by serology is much higher than that predicted by frequent prospective assessment of stool samples.

  4. Moellerella wisconsensis, a new genus and species of Enterobacteriaceae found in human stool specimens.

    Science.gov (United States)

    Hickman-Brenner, F W; Huntley-Carter, G P; Saitoh, Y; Steigerwalt, A G; Farmer, J J; Brenner, D J

    1984-04-01

    The name Moellerella wisconsensis is proposed for a group of the family Enterobacteriaceae previously called enteric group 46. The species name, wisconsensis, was coined because six of the nine strains were isolated in Wisconsin. M. wisconsensis strains were negative for indole production, Voges-Proskauer, H2S production, urea, phenylalanine deaminase, lysine and ornithine decarboxylases, arginine dihydrolase, gas production from D-glucose, acid production from trehalose, and motility; the strains were positive for methyl red, citrate (Simmons), and acid production from lactose and raffinose and resistant to colistin. DNAs from five strains of M. wisconsensis were highly related (80 to 93% in reactions assayed on hydroxyapatite at 60 degrees C and 78 to 97% at 75 degrees C) to 32P-labeled DNA of the proposed type strain (CDC 2896-78, ATCC 35017). Labeled DNA from this type strain was only 2 to 32% related (at 60 degrees C) to DNA from 49 strains of named and unnamed species of Enterobacteriaceae. Eight of nine M. wisconsensis strains were isolated from human stool samples. Clinical information on one strain was available, and it was found to be associated with a case of diarrhea. On MacConkey agar, colonies of M. wisconsensis were bright red with precipitated bile around them and thus were indistinguishable from Escherichia coli colonies. Future studies should focus on the isolation of this new organism and its relationship to human disease.

  5. A safety assessment of rotary mode core sampling in flammable gas single shell tanks: Hanford Site, Richland, Washington

    Energy Technology Data Exchange (ETDEWEB)

    Raymond, R.E.

    1996-04-15

    This safety assessment (SA) addresses each of the required elements associated with the installation, operation, and removal of a rotary-mode core sampling (RMCS) device in flammable-gas single-shell tanks (SSTs). The RMCS operations are needed in order to retrieve waste samples from SSTs with hard layers of waste for which push-mode sampling is not adequate for sampling. In this SA, potential hazards associated with the proposed action were identified and evaluated systematically. Several potential accident cases that could result in radiological or toxicological gas releases were identified and analyzed and their consequences assessed. Administrative controls, procedures and design changes required to eliminate or reduce the potential of hazards were identified. The accidents were analyzed under nine categories, four of which were burn scenarios. In SSTS, burn accidents result in unacceptable consequences because of a potential dome collapse. The accidents in which an aboveground burn propagates into the dome space were shown to be in the ``beyond extremely unlikely`` frequency category. Given the unknown nature of the gas-release behavior in the SSTS, a number of design changes and administrative controls were implemented to achieve these low frequencies. Likewise, drill string fires and dome space fires were shown to be very low frequency accidents by taking credit for the design changes, controls, and available experimental and analytical data. However, a number of Bureau of Mines (BOM) tests must be completed before some of the burn accidents can be dismissed with high confidence. Under the category of waste fires, the possibility of igniting the entrapped gases and the waste itself were analyzed. Experiments are being conducted at the BOM to demonstrate that the drill bit is not capable of igniting the trapped gas in the waste. Laboratory testing and thermal analysis demonstrated that, under normal operating conditions, the drill bit will not create high

  6. A safety assessment of rotary mode core sampling in flammable gas single shell tanks: Hanford Site, Richland, Washington

    International Nuclear Information System (INIS)

    Raymond, R.E.

    1996-01-01

    This safety assessment (SA) addresses each of the required elements associated with the installation, operation, and removal of a rotary-mode core sampling (RMCS) device in flammable-gas single-shell tanks (SSTs). The RMCS operations are needed in order to retrieve waste samples from SSTs with hard layers of waste for which push-mode sampling is not adequate for sampling. In this SA, potential hazards associated with the proposed action were identified and evaluated systematically. Several potential accident cases that could result in radiological or toxicological gas releases were identified and analyzed and their consequences assessed. Administrative controls, procedures and design changes required to eliminate or reduce the potential of hazards were identified. The accidents were analyzed under nine categories, four of which were burn scenarios. In SSTS, burn accidents result in unacceptable consequences because of a potential dome collapse. The accidents in which an aboveground burn propagates into the dome space were shown to be in the ''beyond extremely unlikely'' frequency category. Given the unknown nature of the gas-release behavior in the SSTS, a number of design changes and administrative controls were implemented to achieve these low frequencies. Likewise, drill string fires and dome space fires were shown to be very low frequency accidents by taking credit for the design changes, controls, and available experimental and analytical data. However, a number of Bureau of Mines (BOM) tests must be completed before some of the burn accidents can be dismissed with high confidence. Under the category of waste fires, the possibility of igniting the entrapped gases and the waste itself were analyzed. Experiments are being conducted at the BOM to demonstrate that the drill bit is not capable of igniting the trapped gas in the waste. Laboratory testing and thermal analysis demonstrated that, under normal operating conditions, the drill bit will not create high

  7. [Microsporidian spores in the stool specimens of toddlers, with or without diarrhea, from Tucumán, Argentina].

    Science.gov (United States)

    Valperga, S M; de Jogna Prat, S A; de Valperga, G J; Lazarte, S G; de Trejo, A V; Díaz, N; Hüttman, H M

    1999-01-01

    An investigation has been carried out from September 1995 to December 1997 to search for microsporidian spores in the stool specimens of 344 toddlers aged 1 to 24 months, hospitalized at a pediatric institution in Tucumán. They were classified in two groups: I, made up of 222 children suffering from severe diarrheas, and II by 122 affected by different pathologies, except gastroenteritis. The detection of microsporidia was done by light microscopy in smears of stained stool specimens by using the Weber modified Kokoskin method. Copro-parasitological and coprobacteriological studies were also carried out and the nutritional status of each child was determined. In group I, microsporidia were found in 12/122 cases (7.2%), 4/68 belong to eutrophic children (5.9%), and 12/137 to undernourished children (8.8%); 8/16 positives were found to be related with other enteropatogenics. In group II, microsporidia were detected in 10/122 (8.2%), 4/47 in eutrophic children (8.5%), 4/54 in undernourished children (7.4%) and without data in two cases. They were related with other enteropatogenics in 5/10 positives. Tucumán can be estimated as an area with a low rate of HIV infection in toddlers, then it can be estimated that the studied sample was essentially HIV negative. The occurrence of microsporidia was important and did not show significant differences between toddlers with or without diarrhea, eutrophic or undernourished children.

  8. [Detection of stool antigens of Echinococcus granulosus in dogs belonging to slaughterhouse workers and offal merchants in Metropolitan Lima].

    Science.gov (United States)

    Merino, Veronika; Falcón, Néstor; Morel, Noelia; González, Gualberto

    2017-04-20

    To demonstrate the presence of Echinoccocus granulosus in the definitive host in the city of Lima, Perú, by detecting parasite antigens in the stool of dogs belonging to offal handlers and merchants in authorized slaughterhouses in Metropolitan Lima. Stool samples were collected from 58 dogs and examined using the coproELISA technique for the detection of secretory/excretory antigens of E. granulosus. A survey was conducted to obtain information on pet feeding and handling practices. Positivity to E. granulosus was detected in 13.8% (8/58) of the dogs. In 27.8% (5/18) of the homes, at least one animal showed positivity, and in families that had more than four dogs the chances of finding positivity in at least one dog were higher (P dog tested positive the pets were fed on offal. Of study participants, 94.4% (17) knew nothing about the routes of transmission of hydatid disease. Results show the presence of definitive hosts in the urban area of Lima and underscore the need to more widely disseminate practices for the prevention of parasite transmission.

  9. The positivity of Helicobacter pylori Stool Antigen in patients with Hyperemesis gravidarum.

    Science.gov (United States)

    Bezircioğlu, Incim; Elveren, Hatice Barın; Baloğlu, Ali; Biçer, Merve

    2011-01-01

    We aimed to investigate the possible association between Helicobacter pylori infection and Hyperemesis gravidarum. Thirty-six pregnant women with Hyperemesis gravidarum with severe vomiting (more than 4 times a day), weight loss (≥3 kg), ketonuria and 36 pregnant women gestational age-matched, without nausea and vomiting attending our outpatient clinic for antenatal care were enrolled the study. Demographic data of the patients were registered. Blood samples for hemogram, serum electrolytes (sodium, potassium, chloride, and calcium), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatine, thyroid stimulating hormone (TSH), free T3-T4, total T3-T4, and urine samples for ketonuria, stool samples for HpSA were studied. The data of both groups were compared. Eight Hyperemesis gravidarum patients (22.2%) and 1 control patient (2.8%) were established HpSA positive and it was statistically significant (p:0.037). There was no significant difference between Hyperemesis gravidarum and control subjects in terms of age, gestational week, parity, educational level, socioeconomic status and smoking. There was anemia in 5 Hyperemesis gravidarum patients, 4 of them were HpSA positive. HpSA positivity was more prevalent in Hyperemesis gravidarum patients with anemia (p=0.003). Severe vomiting (more than 4 times a day), heartburn, epigastric pain, duration of hospitalization (more than 4 days) and weight loss (≥5 kg) were not correlated to HpSA positivity. The pregnant women with Hyperemesis gravidarum have a significantly higher prevalence of Helicobacter pylori compared with control subjects.

  10. Prevalence of intestinal protozoa infection among school-aged children on Pemba Island, Tanzania, and effect of single-dose albendazole, nitazoxanide and albendazole-nitazoxanide.

    Science.gov (United States)

    Speich, Benjamin; Marti, Hanspeter; Ame, Shaali M; Ali, Said M; Bogoch, Isaac I; Utzinger, Jürg; Albonico, Marco; Keiser, Jennifer

    2013-01-04

    Pathogenic intestinal protozoa infections are common in school-aged children in the developing world and they are frequently associated with malabsorption syndromes and gastrointestinal morbidity. Since diagnosis of these parasites is difficult, prevalence data on intestinal protozoa is scarce. We collected two stool samples from school-aged children on Pemba Island, Tanzania, as part of a randomized controlled trial before and 3 weeks after treatment with (i) single-dose albendazole (400 mg); (ii) single-dose nitazoxanide (1,000 mg); (iii) nitazoxanide-albendazole combination (1,000 mg-400 mg), with each drug given separately on two consecutive days; and (iv) placebo. Formalin-fixed stool samples were examined for the presence of intestinal protozoa using an ether-concentration method to determine the prevalence and estimate cure rates (CRs). Almost half (48.7%) of the children were diagnosed with at least one of the (potentially) pathogenic protozoa Giardia intestinalis, Entamoeba histolytica/E. dispar and Blastocystis hominis. Observed CRs were high for all treatment arms, including placebo. Nitazoxanide showed a significant effect compared to placebo against the non-pathogenic protozoon Entamoeba coli. Intestinal protozoa infections might be of substantial health relevance even in settings where they are not considered as a health problem. Examination of a single stool sample with the ether-concentration method lacks sensitivity for the diagnosis of intestinal protozoa, and hence, care is indicated when interpreting prevalence estimates and treatment effects.

  11. Searching for stable Na-ordered phases in single-crystal samples of γ-NaxCoO2

    Science.gov (United States)

    Shu, G. J.; Prodi, Andrea; Chu, S. Y.; Lee, Y. S.; Sheu, H. S.; Chou, F. C.

    2007-11-01

    We report on the preparation and characterization of single-crystal γ phase NaxCoO2 with 0.25≤x≤0.84 using a nonaqueous electrochemical chronoamperemetry technique. By carefully mapping the overpotential versus x (for xfind six distinct stable phases with Na levels corresponding to xtilde 0.75, 0.71, 0.50, 0.43, 0.33, and 0.25. The composition with x≃0.55 appears to have a critical Na concentration which separates samples with different magnetic behavior as well as different Na ion diffusion mechanisms. Chemical analysis of an aged crystal reveals different Na ion diffusion mechanisms above and below xc˜0.53 , where the diffusion process above xc has a diffusion coefficient about five times larger than that below xc . The series of crystals were studied with x-ray diffraction, susceptibility, and transport measurements. The crystal with x=0.5 shows a weak ferromagnetic transition below T=27K in addition to the usual transitions at T=51 and 88K . The resistivity of the Curie-Weiss metallic Na0.71CoO2 composition has a very low residual resistivity, which attests to the high homogeneity of the crystals prepared by this improved electrochemical method. Our results on the various stable crystal compositions point to the importance of Na ion ordering across the phase diagram.

  12. A device for the application of uniaxial strain to single crystal samples for use in synchrotron radiation experiments

    Energy Technology Data Exchange (ETDEWEB)

    Gannon, L. [Clarendon Laboratory, University of Oxford Physics Department, Parks Road, Oxford OX1 3PU (United Kingdom); Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 ODE (United Kingdom); Bosak, A. [European Synchrotron Radiation Facility, BP 220, F-38043 Grenoble Cedex (France); Burkovsky, R. G. [European Synchrotron Radiation Facility, BP 220, F-38043 Grenoble Cedex (France); Peter the Great Saint-Petersburg Polytechnic University, 29 Politekhnicheskaya, 195251, St.-Petersburg (Russian Federation); Nisbet, G.; Hoesch, M., E-mail: Moritz.Hoesch@diamond.ac.uk [Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 ODE (United Kingdom); Petrović, A. P. [DPMC-MaNEP, Université de Genève, Quai Ernest-Ansermet 24, 1211 Genève 4 (Switzerland)

    2015-10-15

    We present the design, construction, and testing of a straining device compatible with many different synchrotron radiation techniques, in a wide range of experimental environments (including low temperature, high field and ultra-high vacuum). The device has been tested by X-ray diffraction on single crystal samples of quasi-one-dimensional Cs{sub 2}Mo{sub 6}Se{sub 6} and K{sub 2}Mo{sub 6}Se{sub 6}, in which microscopic strains up to a Δc/c = 0.12% ± 0.01% change in the c lattice parameters have been achieved. We have also used the device in an inelastic X-ray scattering experiment, to probe the strain-dependent speed of sound ν along the c axis. A reduction Δν/ν of up to −3.8% was obtained at a strain of Δc/c = 0.25% in K{sub 2}Mo{sub 6}Se{sub 6}.

  13. A disposable screen-printed silver strip sensor for single drop analysis of halide in biological samples.

    Science.gov (United States)

    Chiu, Mei-Hsin; Cheng, Wan-Ling; Muthuraman, Govindan; Hsu, Cheng-Teng; Chung, Hsieh-Hsun; Zen, Jyh-Myng

    2009-06-15

    A screen-printed silver strip with three-electrode configuration of Ag-working, Ag-counter and Ag/Ag(x)O reference electrodes was developed for simultaneous determination of chloride, bromide and iodide in aqueous solutions. It was fabricated simply by screen-printing silver ink onto a polypropylene (PP) base. The in-situ prepared Ag/Ag(x)O reference electrode can avoid the leaching interference in chloride detection while using a conventional Ag/AgCl reference electrode. A single drop of analyte (50 microl) is enough to determine iodide, bromide and chloride by measuring the well-separated oxidation peak currents of respective silver halides. The calibration graph was linear from 10 microM to 20 mM for iodide and bromide and 100 microM to 20 mM for chloride and the detection limit (S/N=3) was 3.05 microM, 2.95 microM and 18.83 microM for iodide, bromide and chloride, respectively. The strip is designed to be disposable and as such manual polishing is not necessary. The proposed sensor is not only simple to manufacture and easy to operate but also fast and precise with little detection volume. It is successfully applied to the determination of halide ions in real samples.

  14. LUMINEX®: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools.

    Science.gov (United States)

    Santos, Helena Lúcia Carneiro; Bandyopadhyay, Kakali; Bandea, Rebecca; Peralta, Regina Helena Saramago; Peralta, José Mauro; Da Silva, Alexandre Januário

    2013-03-15

    Six species of the genus Entamoeba, i.e., E. histolytica, E. dispar, E. moshkovskii, E. polecki, E. coli, and E. hartmanii can be found in human stools. Among these, only E. histolytica is considered to be pathogenic, causing intestinal and extra-intestinal disease, but it is morphologically identical to E. dispar and E. moshkovskii. In general, E. polecki, E. coli, and E. hartmanii can be differentiated morphologically from E. histolytica, but some of their diagnostic morphologic features may overlap creating issues for the differential diagnosis. Moreover, the previous inability to differentiate among Entamoeba species has limited epidemiologic information on E histolytica. The objective of this study was to develop a rapid, high-throughput screening method using Luminex technique for the simultaneous detection and differentiation of Entamoeba species. PCR amplification was performed with biotinylated Entamoeba sp 18S rRNA gene primers, designed to amplify a fragment ranging from 382 to 429 bp of the Entamoeba spp studied. Regions of this fragment that could differentiate among E. histolytica, E. moshkovskii, E. dispar, E. hartmanii and E. coli were selected to design hybridization probes to link to Luminex beads. The assay was standardized with cloned DNA samples of each species and evaluated with 24 DNA extracts from samples obtained from individuals diagnosed with these amebas in their stools. Using this approach we were able to correctly identify E. histoltyica, E. dispar, E hartmanni, E. coli and E. moshkovskii in all specimens studied. From twenty four samples tested by microscopy, PCR/DNA Sequencing and real-time PCR, 100% agreed with PCR-Luminex assay for identification of E. dispar, E. moshkovskii, E. hartmanni, E. histolytica, and E. coli. These results show that this method could be used in the diagnostic detection of Entamoeba spp in fecal samples. This diagnostic test was useful to clearly distinguish E histolytica from other species and also to

  15. Modification of stool's water content in constipated infants: management with an adapted infant formula

    Directory of Open Access Journals (Sweden)

    Alvarez Marina M

    2011-05-01

    Full Text Available Abstract Background Constipation is a common occurrence in formula-fed infants. The aim of this preliminary study was to evaluate the impact of a formula with high levels of lactose and magnesium, in compliance with the official regulations, on stool water content, as well as a parental assessment of constipation. Materials and methods Thirty healthy term-born, formula-fed infants, aged 4-10 weeks, with functional constipation were included. All infants were full-term and fed standard formula. Exclusion criteria were preterm and/or low birth weight, organic constipation, being breast fed or fed a formula specially designed to treat constipation. Stool composition was measured by near-infrared reflectance analysis (NIRA and parents answered questions about crying associated with defecation and stool consistency at baseline and after two weeks of the adapted formula. Results After 2 weeks of the adapted formula, stool water content increased from 71 +/- 8.1% to 84 +/- 5.9%, (p Conclusions This preliminary study suggests that an adapted formula with high levels of lactose and magnesium increases stool water content and improves symptoms of constipation in term-born, formula-fed infants. A larger randomized placebo-controlled trial is indicated.

  16. Detection of Giardia lamblia and Cryptosporidium parvum by direct immunofluorescence assay in stool specimen.

    Science.gov (United States)

    Rahman, M M; Hossain, M A; Paul, S K; Ahmed, S; Islam, A; Ehsan, M A; Alam, M M; Kabir, M R; Sarkar, S R

    2014-07-01

    Giardia and Cryptosporidium are the pathogens which transmitted through contaminated soil and contaminated water are significant causes of diarrhea and nutritional disorders in institutional and community peoples. Children and immune compromise persons are more vulnerable for these infections. Both Giardiasis and Cryptosporidiosis were included in 2004 as WHO Neglected Disease. So this is a major public health problem in developing countries. The present study was carried out to detect the Giardia and Cryptosporidium from diarrheic or patient having loose stool by Direct Immunofluorescence assay. The study was conducted during July 20012 to February 2013 and the work was done in Mymensingh Medical College in the department of Microbiology and in Bangladesh Agricultural University in the department of Veterinary Medicine. A total of 100 loose stools were collected from school children of different area and hospital under sadar upazilla, Mymensingh. The detection of Giardia lamblia and Cryptosporidium parvum showed the individual prevalence 8% and 4% respectively. The highest cyst/oocyst count was 85,000 and 1,000/gm of stool and the lowest being 100 and 50/gm of stool for Giardiasis and Cryptosporidiosis respectively. The detection rate of Giardia and Cryptosporidium by Immunofluorescence assay was relatively higher than the previous study done in Bangladesh and this was the first report from Bangladesh over human stool specimen using Immunofluorescence assay. So, Immunofluorescence assay could be adapted for rapid and accurate detection of Giardia and Cryptosporidium.

  17. Picky eating in preschool children: Associations with dietary fibre intakes and stool hardness

    Science.gov (United States)

    Taylor, Caroline M.; Northstone, Kate; Wernimont, Susan M.; Emmett, Pauline M.

    2018-01-01

    It has been suggested that constipation may be associated with picky eating. Constipation is a common condition in childhood and a low intake of dietary fibre may be a risk factor. Differences in fibre intake between picky and non-picky children and its relation to stool consistency is currently not well-understood. Children enrolled in the Avon Longitudinal Study of Parents and Children identified as picky eaters (PE) were compared with non-picky eaters (NPE): (1) to determine dietary fibre intake at 38 months; (2) to investigate whether any difference in dietary fibre intake was predictive of usual stool hardness at 42 months. PE was identified from questionnaires at 24 and 38 months. Usual stool hardness was identified from a questionnaire at 42 months. Dietary intake was assessed at 38 months with a food frequency questionnaire. Dietary fibre intake was lower in PE than NPE (mean difference −1.4 (95% CI −1.6, −1.2) g/day, p children was associated with an increased prevalence of usually having hard stools. This association was mediated by low dietary fibre intake, particularly from vegetables, in PE. For children with PE, dietary advice aimed at increasing fibre intake may help avoid hard stools. PMID:26879221

  18. Measuring word complexity in speech screening: single-word sampling to identify phonological delay/disorder in preschool children.

    Science.gov (United States)

    Anderson, Carolyn; Cohen, Wendy

    2012-01-01

    Children's speech sound development is assessed by comparing speech production with the typical development of speech sounds based on a child's age and developmental profile. One widely used method of sampling is to elicit a single-word sample along with connected speech. Words produced spontaneously rather than imitated may give a more accurate indication of a child's speech development. A published word complexity measure can be used to score later-developing speech sounds and more complex word patterns. There is a need for a screening word list that is quick to administer and reliably differentiates children with typically developing speech from children with patterns of delayed/disordered speech. To identify a short word list based on word complexity that could be spontaneously named by most typically developing children aged 3;00-5;05 years. One hundred and five children aged between 3;00 and 5;05 years from three local authority nursery schools took part in the study. Items from a published speech assessment were modified and extended to include a range of phonemic targets in different word positions in 78 monosyllabic and polysyllabic words. The 78 words were ranked both by phonemic/phonetic complexity as measured by word complexity and by ease of spontaneous production. The ten most complex words (hereafter Triage 10) were named spontaneously by more than 90% of the children. There was no significant difference between the complexity measures for five identified age groups when the data were examined in 6-month groups. A qualitative analysis revealed eight children with profiles of phonological delay or disorder. When these children were considered separately, there was a statistically significant difference (p speech from those with delayed or disordered speech patterns. The Triage 10 words can be used as a screening tool for triage and general assessment and have the potential to monitor progress during intervention. Further testing is being undertaken to

  19. A generic-tee-plenum mixing system for application to single point aerosol sampling in stacks and ducts.

    Science.gov (United States)

    Han, Taewon; O'Neal, Dennis L; Ortiz, Carlos A

    2007-01-01

    The ANSI/HPS-N13.1-1999 standard is based on the concept of obtaining a single point representative sample from a location where the velocity and contaminant profiles are relatively uniform. It is difficult to predict the level of mixing in an arbitrary stack or duct without experimental data to meet the ANSI/HPS N13.1-1999 requirements. The goal of this study was to develop experimental data for a range of conditions in "S" (S-shaped configuration) duct systems with different mixing elements and "S" systems having one or two mixing elements. Results were presented in terms of the coefficients of variation (COVs) for velocity, tracer gas, and 10-mum aerodynamic diameter (AD) aerosol particle profiles at different downstream locations for each mixing element. Five mixing elements were tested, including a 90 degrees elbow, a commercial static mixer, a Small-Horizontal Generic-Tee-Plenum (SH-GTP), a Small-Vertical Generic-Tee-Plenum (SV-GTP), and a Large-Horizontal Generic-Tee-Plenum (LH-GTP) system. The COVs for velocity, gas concentration, and aerosol particles for the three GTP systems were all determined to be less than 8%. Tests with two different sizes of GTPs were conducted, and the results showed the performance of the GTPs was relatively unaffected by either size or velocity as reflected by the Reynolds number. The pressure coefficients were 0.59, 0.57, and 0.65, respectively, for the SH-GTP, SV-GTP, and LH-GTP. The pressure drop for the GTPs was approximately twice that of the round elbow, but a factor of 5 less than a Type IV Air Blender. The GTP was developed to provide a sampling location less than 4-duct diameters downstream of a mixing element with low pressure drop condition. The object of the developmental effort was to provide a system that could be employed in new stack; however, the concept of GTPs could also be retrofitted onto existing system applications as well. Results from these tests show that the system performance is well within the ANSI

  20. Feasibility study of a single-shot 3D electron bunch shape monitor with an electro-optic sampling technique

    Directory of Open Access Journals (Sweden)

    Yuichi Okayasu

    2013-05-01

    Full Text Available We developed a three-dimensional electron bunch charge distribution (3D-BCD monitor with single-shot detection, and a spectral decoding based electro-optic (EO sampling technique for a nondestructive monitor enables real-time reconstruction of the three-dimensional distribution of a bunch charge. We realized three goals by simultaneously probing a number of Pockels EO crystals that surround the electron beam axis with hollow and radial polarized laser pulses. First, we performed a feasibility test as a simple case of a 3D-BCD monitor probing two ZnTe crystals as EO detectors installed on the opposite angle to the electron beam axis and confirmed that we simultaneously obtained both EO signals. Since the adopted hollow probe laser pulse is not only radially polarized but also temporally shifted azimuthally, some disorders in the radial polarization distribution of such a laser pulse were numerically analyzed with a plane-wave expansion method. Based on the above investigations, the 3D-BCD monitor is feasible both in experimental and numerical estimations. Furthermore, we previously developed a femtosecond response organic crystal as a Pockels EO detector and a broadband probe laser (≥350  nm in FWHM; the 3D-BCD monitor realizes 30- to 40-fs (FWHM temporal resolution. Eventually, the monitor is expected to be equipped in such advanced accelerators as XFEL to measure and adjust the electron bunch charge distribution in real time. The 3D-BCD measurement works as a critical tool to provide feedback to seeded FELs.

  1. Yield of Stool Culture with Isolate Toxin Testing versus a Two-Step Algorithm Including Stool Toxin Testing for Detection of Toxigenic Clostridium difficile▿

    Science.gov (United States)

    Reller, Megan E.; Lema, Clara A.; Perl, Trish M.; Cai, Mian; Ross, Tracy L.; Speck, Kathleen A.; Carroll, Karen C.

    2007-01-01

    We examined the incremental yield of stool culture (with toxin testing on isolates) versus our two-step algorithm for optimal detection of toxigenic Clostridium difficile. Per the two-step algorithm, stools were screened for C. difficile-associated glutamate dehydrogenase (GDH) antigen and, if positive, tested for toxin by a direct (stool) cell culture cytotoxicity neutralization assay (CCNA). In parallel, stools were cultured for C. difficile and tested for toxin by both indirect (isolate) CCNA and conventional PCR if the direct CCNA was negative. The “gold standard” for toxigenic C. difficile was detection of C. difficile by the GDH screen or by culture and toxin production by direct or indirect CCNA. We tested 439 specimens from 439 patients. GDH screening detected all culture-positive specimens. The sensitivity of the two-step algorithm was 77% (95% confidence interval [CI], 70 to 84%), and that of culture was 87% (95% CI, 80 to 92%). PCR results correlated completely with those of CCNA testing on isolates (29/29 positive and 32/32 negative, respectively). We conclude that GDH is an excellent screening test and that culture with isolate CCNA testing detects an additional 23% of toxigenic C. difficile missed by direct CCNA. Since culture is tedious and also detects nontoxigenic C. difficile, we conclude that culture is most useful (i) when the direct CCNA is negative but a high clinical suspicion of toxigenic C. difficile remains, (ii) in the evaluation of new diagnostic tests for toxigenic C. difficile (where the best reference standard is essential), and (iii) in epidemiologic studies (where the availability of an isolate allows for strain typing and antimicrobial susceptibility testing). PMID:17804652

  2. Yield of stool culture with isolate toxin testing versus a two-step algorithm including stool toxin testing for detection of toxigenic Clostridium difficile.

    Science.gov (United States)

    Reller, Megan E; Lema, Clara A; Perl, Trish M; Cai, Mian; Ross, Tracy L; Speck, Kathleen A; Carroll, Karen C

    2007-11-01

    We examined the incremental yield of stool culture (with toxin testing on isolates) versus our two-step algorithm for optimal detection of toxigenic Clostridium difficile. Per the two-step algorithm, stools were screened for C. difficile-associated glutamate dehydrogenase (GDH) antigen and, if positive, tested for toxin by a direct (stool) cell culture cytotoxicity neutralization assay (CCNA). In parallel, stools were cultured for C. difficile and tested for toxin by both indirect (isolate) CCNA and conventional PCR if the direct CCNA was negative. The "gold standard" for toxigenic C. difficile was detection of C. difficile by the GDH screen or by culture and toxin production by direct or indirect CCNA. We tested 439 specimens from 439 patients. GDH screening detected all culture-positive specimens. The sensitivity of the two-step algorithm was 77% (95% confidence interval [CI], 70 to 84%), and that of culture was 87% (95% CI, 80 to 92%). PCR results correlated completely with those of CCNA testing on isolates (29/29 positive and 32/32 negative, respectively). We conclude that GDH is an excellent screening test and that culture with isolate CCNA testing detects an additional 23% of toxigenic C. difficile missed by direct CCNA. Since culture is tedious and also detects nontoxigenic C. difficile, we conclude that culture is most useful (i) when the direct CCNA is negative but a high clinical suspicion of toxigenic C. difficile remains, (ii) in the evaluation of new diagnostic tests for toxigenic C. difficile (where the best reference standard is essential), and (iii) in epidemiologic studies (where the availability of an isolate allows for strain typing and antimicrobial susceptibility testing).

  3. Use of string test and stool specimens to diagnose pulmonary tuberculosis.

    Science.gov (United States)

    DiNardo, Andrew R; Hahn, Andrew; Leyden, Jacinta; Stager, Charles; Jo Baron, Ellen; Graviss, Edward A; Mandalakas, Anna M; Guy, Elizabeth

    2015-12-01

    The Xpert MTB/RIF (MTB/RIF) test has advanced the field of tuberculosis (TB) diagnostics; however, depending on age and HIV status, 10-85% of individuals with presumed pulmonary TB (PTB) are unable to produce sputum. The feasibility of using MTB/RIF and culture on stool and string test specimens from 13 adult patients with presumed PTB was studied. The string test was well tolerated with a median Wong Baker Faces score of 2. The string test had 100% sensitivity and specificity by MTB/RIF and 87.5% sensitivity and 100% specificity by culture. In stool, Mycobacterium tuberculosis DNA was detected in all cases of culture-confirmed PTB. The string test and stool provide diagnostic specimens that warrant further investigation. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  4. Evaluation of a PCR/DNA Probe Colorimetric Membrane Assay for Identification of Campylobacter spp. in Human Stool Specimens

    OpenAIRE

    Collins, Evelyn; Glennon, Maura; Hanley, Shirley; Murray, Anne-Marie; Cormican, Martin; Smith, Terry; Maher, Majella

    2001-01-01

    DNA was extracted from 50 human stool specimens using the QIAamp DNA stool minikit. PCR amplification was followed by post-PCR hybridization to DNA probes specific for the Campylobacter genus, Campylobacter jejuni, and Campylobacter coli in a colorimetric membrane assay. Thirty-two of 38 culture-positive specimens were PCR/DNA probe positive for C. jejuni. The assay is rapid and simple and can be applied to stool specimens for the detection of Campylobacter.

  5. On the preparation of as-produced and purified single-walled carbon nanotube samples for standardized X-ray diffraction characterization

    International Nuclear Information System (INIS)

    Allaf, Rula M.; Rivero, Iris V.; Spearman, Shayla S.; Hope-Weeks, Louisa J.

    2011-01-01

    The aim of this research was to specify proper sample conditioning for acquiring representative X-ray diffraction (XRD) profiles for single-walled carbon nanotube (SWCNT) samples. In doing so, a specimen preparation method for quantitative XRD characterization of as-produced and purified arc-discharge SWCNT samples has been identified. Series of powder XRD profiles were collected at different temperatures, states, and points of time to establish appropriate conditions for acquiring XRD profiles without inducing much change to the specimen. It was concluded that heating in the 300-450 deg. C range for 20 minutes, preferably vacuum-assisted, and then sealing the sample is an appropriate XRD specimen preparation technique for purified arc-discharge SWCNT samples, while raw samples do not require preconditioning for characterization. - Graphical Abstract: A sample preparation method for XRD characterization of as-produced and purified arc-discharge SWCNT samples is identified. The preparation technique seeks to acquire representative XRD profiles without inducing changes to the samples. Purified samples required 20 minutes of heating at (300-450)deg. C, while raw samples did not require preconditioning for characterization. Highlights: → Purification routines may induce adsorption onto the SWCNT samples. → Heating a SWCNT sample may result in material loss, desorption, and SWCNTs closing. → Raw arc-discharge samples do not require preparation for XRD characterization. → Heating is appropriate specimen preparation for purified and heat-treated samples. → XRD data fitting is required for structural analysis of SWCNT bundles.

  6. Determination of the technetium-99m mercaptoacetyltriglycine plasma clearance in children by means of a single blood sample: a multicentre study

    International Nuclear Information System (INIS)

    Piepsz, A.; Gordon, I.; Hahn, K.; Kolinska, J.; Kotzerke, J.; Sixt, R.

    1993-01-01

    A multicentree European study was undertaken in order to determine a reasonable algorithm allowing the determination of overall technetium-99m mercaptoacetyltriglycine clearance using a single blood sample. Employing multiple blood sample clearance as a reference method, it was shown that an acceptable estimation of the MAG3 renal clearance could be obtained using a blood sample taken at any time between 30 and 40 min after tracer injection. After correction for body surface area comparison of clearance determined using (a) the single blood sample and (b) the multiple blood samples provided a coefficient of correlation of 0.949 and an SEE of 27 ml/min. This algorithm is valid for clearance values higher than 100 ml/min/1.73 m 2 and for children older than 1 year of age. (orig.)

  7. Sample Preparation Methods Following CellSearch Approach Compatible of Single-Cell Whole-Genome Amplification: An Overview

    NARCIS (Netherlands)

    Swennenhuis, Joost Franciscus; Terstappen, Leonardus Wendelinus Mathias Marie; Kroneis, Thomas

    2015-01-01

    Single cells are increasingly used to determine the heterogeneity of therapy targets in the genome during the course of a disease. The first challenge using single cells is to isolate these cells from the surrounding cells, especially when the targeted cells are rare. A number of techniques have

  8. Use of Bristol Stool Form Scale to predict the adequacy of bowel preparation - a prospective study.

    Science.gov (United States)

    Malhotra, A; Shah, N; Depasquale, J; Baddoura, W; Spira, R; Rector, T

    2016-02-01

    Inadequate bowel preparation continues to be a substantial problem for colonoscopy. The seven-point Bristol Stool Form Scale (BSFS) has been associated with delayed colonic transit in adults. We evaluated the utility of the BSFS to identify patients more likely to present with an inadequate preparation. Two large community-based academic medical centres in New Jersey, USA, studied a prospective cohort of 411 consecutive patients undergoing outpatient colonoscopy who were prescribed similar bowel preparations. The BSFS and several other study variables were collected by gastroenterology fellows during an outpatient visit prior to scheduling colonoscopy. All colonoscopy examinations were performed in the morning by a gastroenterologist who graded the adequacy of bowel preparation. Inadequate preparation was defined as one resulting in a repeat colonoscopy at a shorter time interval than would generally be recommended based solely on risk factors or pathological findings. The ability of study variables to discriminate those who did or did not have an adequate preparation was summarized by the c-statistic. The relationship between variables that provided some discrimination and the probability of an adequate preparation was modelled using logistic regression. The mean age of the study sample was 56 ± 8 (SD) years and 63% were women. Bowel preparation was adequate in 337 (82%) of the patients. The BSFS ratings ranged from 1 to 7. The score was <3 in 144 (35%) indicating lower gastrointestinal motility. There was a statistically significant association between the score and the probability of an adequate bowel preparation (odds ratio 1.4; 95% confidence interval 1.2-1.7; P < 0.001) and the c-statistic was 0.64 (0.58-0.70). Use of the BSFS may help identify patients for whom standard bowel preparation most probably will not be adequate. Colorectal Disease © 2015 The Association of Coloproctology of Great Britain and Ireland.

  9. Testing single-grain quartz OSL methods using sediment samples with independent age control from the Bordes-Fitte rockshelter (Roches d'Abilly site, Central France)

    DEFF Research Database (Denmark)

    Thomsen, Kristina Jørkov; Murray, Andrew Sean; Buylaert, Jan-Pieter

    2016-01-01

    We present quartz single-grain dose distributions for four well-bleached and unmixed sediment samples with independent age control (22–48 ka), from the archaeologically important Bordes-Fitte rockshelter at Roches d'Abilly, France. This site has previously been dated using 14C AMS dating and stan......We present quartz single-grain dose distributions for four well-bleached and unmixed sediment samples with independent age control (22–48 ka), from the archaeologically important Bordes-Fitte rockshelter at Roches d'Abilly, France. This site has previously been dated using 14C AMS dating...

  10. Estimating the sensitivity and specificity of Kato-Katz stool examination technique for detection of hookworms, Ascaris lumbricoides and Trichuris trichiura infections in humans in the absence of a 'gold standard'.

    Science.gov (United States)

    Tarafder, M R; Carabin, H; Joseph, L; Balolong, E; Olveda, R; McGarvey, S T

    2010-03-15

    The accuracy of the Kato-Katz technique in identifying individuals with soil-transmitted helminth (STH) infections is limited by day-to-day variation in helminth egg excretion, confusion with other parasites and the laboratory technicians' experience. We aimed to estimate the sensitivity and specificity of the Kato-Katz technique to detect infection with Ascaris lumbricoides, hookworm and Trichuris trichiura using a Bayesian approach in the absence of a 'gold standard'. Data were obtained from a longitudinal study conducted between January 2004 and December 2005 in Samar Province, the Philippines. Each participant provided between one and three stool samples over consecutive days. Stool samples were examined using the Kato-Katz technique and reported as positive or negative for STHs. In the presence of measurement error, the true status of each individual is considered as latent data. Using a Bayesian method, we calculated marginal posterior densities of sensitivity and specificity parameters from the product of the likelihood function of observed and latent data. A uniform prior distribution was used (beta distribution: alpha=1, beta=1). A total of 5624 individuals provided at least one stool sample. One, two and three stool samples were provided by 1582, 1893 and 2149 individuals, respectively. All STHs showed variation in test results from day to day. Sensitivity estimates of the Kato-Katz technique for one stool sample were 96.9% (95% Bayesian Credible Interval [BCI]: 96.1%, 97.6%), 65.2% (60.0%, 69.8%) and 91.4% (90.5%, 92.3%), for A. lumbricoides, hookworm and T. trichiura, respectively. Specificity estimates for one stool sample were 96.1% (95.5%, 96.7%), 93.8% (92.4%, 95.4%) and 94.4% (93.2%, 95.5%), for A. lumbricoides, hookworm and T. trichiura, respectively. Our results show that the Kato-Katz technique can perform with reasonable accuracy with one day's stool collection for A. lumbricoides and T. trichiura. Low sensitivity of the Kato-Katz for detection

  11. Effect of single-layer centrifugation or washing on frozen-thawed donkey semen quality: Do they have the same effect regardless of the quality of the sample?

    Science.gov (United States)

    Ortiz, I; Dorado, J; Morrell, J M; Crespo, F; Gosálvez, J; Gálvez, M J; Acha, D; Hidalgo, M

    2015-07-15

    The aims of this study were to determine the sperm quality of frozen-thawed donkey sperm samples after single-layer centrifugation (SLC) using Androcoll-E in comparison to sperm washing or no centrifugation and to determine if the effect on the sperm quality after SLC or sperm washing depends on the quality of the sample. Frozen-thawed sperm samples from Andalusian donkeys were divided into three aliquots, and they were processed using three different techniques after thawing: uncentrifuged diluted control (UDC), sperm washing (SW), and SLC. Afterward, sperm quality index was estimated by integrating all parameters (total and progressive sperm motility, membrane integrity, and DNA fragmentation) in a single value. The relationship between the sperm quality of thawed UDC samples and the effect on sperm parameters in SW and SLC-selected samples was assessed. Sperm quality index was significantly higher (P < 0.001) in SLC (0.8 ± 0.0) samples than that in UDC (0.6 ± 0.0) and SW (0.6 ± 0.0) samples, regardless of the sperm quality index after thawing of the sperm sample. In conclusion, SLC of frozen-thawed donkey spermatozoa using Androcoll-E-Small can be a suitable procedure for selecting frozen-thawed donkey sperm with better quality, in particular in those samples where an improvement in motility is needed. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Providing Quantitative Information and a Nudge to Undergo Stool Testing in a Colorectal Cancer Screening Decision Aid: A Randomized Clinical Trial.

    Science.gov (United States)

    Schwartz, Peter H; Perkins, Susan M; Schmidt, Karen K; Muriello, Paul F; Althouse, Sandra; Rawl, Susan M

    2017-08-01

    Guidelines recommend that patient decision aids should provide quantitative information about probabilities of potential outcomes, but the impact of this information is unknown. Behavioral economics suggests that patients confused by quantitative information could benefit from a "nudge" towards one option. We conducted a pilot randomized trial to estimate the effect sizes of presenting quantitative information and a nudge. Primary care patients (n = 213) eligible for colorectal cancer screening viewed basic screening information and were randomized to view (a) quantitative information (quantitative module), (b) a nudge towards stool testing with the fecal immunochemical test (FIT) (nudge module), (c) neither a nor b, or (d) both a and b. Outcome measures were perceived colorectal cancer risk, screening intent, preferred test, and decision conflict, measured before and after viewing the decision aid, and screening behavior at 6 months. Patients viewing the quantitative module were more likely to be screened than those who did not ( P = 0.012). Patients viewing the nudge module had a greater increase in perceived colorectal cancer risk than those who did not ( P = 0.041). Those viewing the quantitative module had a smaller increase in perceived risk than those who did not ( P = 0.046), and the effect was moderated by numeracy. Among patients with high numeracy who did not view the nudge module, those who viewed the quantitative module had a greater increase in intent to undergo FIT ( P = 0.028) than did those who did not. The limitations of this study were the limited sample size and single healthcare system. Adding quantitative information to a decision aid increased uptake of colorectal cancer screening, while adding a nudge to undergo FIT did not increase uptake. Further research on quantitative information in decision aids is warranted.

  13. Preparation and Loading Process of Single Crystalline Samples into a Gas Environmental Cell Holder for In Situ Atomic Resolution Scanning Transmission Electron Microscopic Observation.

    Science.gov (United States)

    Straubinger, Rainer; Beyer, Andreas; Volz, Kerstin

    2016-06-01

    A reproducible way to transfer a single crystalline sample into a gas environmental cell holder for in situ transmission electron microscopic (TEM) analysis is shown in this study. As in situ holders have only single-tilt capability, it is necessary to prepare the sample precisely along a specific zone axis. This can be achieved by a very accurate focused ion beam lift-out preparation. We show a step-by-step procedure to prepare the sample and transfer it into the gas environmental cell. The sample material is a GaP/Ga(NAsP)/GaP multi-quantum well structure on Si. Scanning TEM observations prove that it is possible to achieve atomic resolution at very high temperatures in a nitrogen environment of 100,000 Pa.

  14. Characterization of bacteria in biopsies of colon and stools by high throughput sequencing of the V2 region of bacterial 16S rRNA gene in human.

    Science.gov (United States)

    Momozawa, Yukihide; Deffontaine, Valérie; Louis, Edouard; Medrano, Juan F

    2011-02-10

    The characterization of the human intestinal microflora and their interactions with the host have been identified as key components in the study of intestinal disorders such as inflammatory bowel diseases. High-throughput sequencing has enabled culture-independent studies to deeply analyze bacteria in the gut. It is possible with this technology to systematically analyze links between microbes and the genetic constitution of the host, such as DNA polymorphisms and methylation, and gene expression. In this study the V2 region of the bacterial 16S ribosomal RNA (rRNA) gene using 454 pyrosequencing from seven anatomic regions of human colon and two types of stool specimens were analyzed. The study examined the number of reads needed to ascertain differences between samples, the effect of DNA extraction procedures and PCR reproducibility, and differences between biopsies and stools in order to design a large scale systematic analysis of gut microbes. It was shown (1) that sequence coverage lower than 1,000 reads influenced quantitative and qualitative differences between samples measured by UniFrac distances. Distances between samples became stable after 1,000 reads. (2) Difference of extracted bacteria was observed between the two DNA extraction methods. In particular, Firmicutes Bacilli were not extracted well by one method. (3) Quantitative and qualitative difference in bacteria from ileum to rectum colon were not observed, but there was a significant positive trend between distances within colon and quantitative differences. Between sample type, biopsies or stools, quantitative and qualitative differences were observed. Results of human colonic bacteria analyzed using high-throughput sequencing were highly dependent on the experimental design, especially the number of sequence reads, DNA extraction method, and sample type.

  15. Characterization of bacteria in biopsies of colon and stools by high throughput sequencing of the V2 region of bacterial 16S rRNA gene in human.

    Directory of Open Access Journals (Sweden)

    Yukihide Momozawa

    Full Text Available BACKGROUND: The characterization of the human intestinal microflora and their interactions with the host have been identified as key components in the study of intestinal disorders such as inflammatory bowel diseases. High-throughput sequencing has enabled culture-independent studies to deeply analyze bacteria in the gut. It is possible with this technology to systematically analyze links between microbes and the genetic constitution of the host, such as DNA polymorphisms and methylation, and gene expression. METHODS AND FINDINGS: In this study the V2 region of the bacterial 16S ribosomal RNA (rRNA gene using 454 pyrosequencing from seven anatomic regions of human colon and two types of stool specimens were analyzed. The study examined the number of reads needed to ascertain differences between samples, the effect of DNA extraction procedures and PCR reproducibility, and differences between biopsies and stools in order to design a large scale systematic analysis of gut microbes. It was shown (1 that sequence coverage lower than 1,000 reads influenced quantitative and qualitative differences between samples measured by UniFrac distances. Distances between samples became stable after 1,000 reads. (2 Difference of extracted bacteria was observed between the two DNA extraction methods. In particular, Firmicutes Bacilli were not extracted well by one method. (3 Quantitative and qualitative difference in bacteria from ileum to rectum colon were not observed, but there was a significant positive trend between distances within colon and quantitative differences. Between sample type, biopsies or stools, quantitative and qualitative differences were observed. CONCLUSIONS: Results of human colonic bacteria analyzed using high-throughput sequencing were highly dependent on the experimental design, especially the number of sequence reads, DNA extraction method, and sample type.

  16. Use of stool culture as a non invasive method for the diagnosis of ...

    African Journals Online (AJOL)

    Background: Helicobacter pylori has been associated with chronic diarrhoea, iron deficiency anaemia, growth retardation, gastric malignancies, peptic ulcer disease, and gastritis among children. Diagnosis of this infection has been invasive using biopsies while stool culture is not common or routinely practiced. This study ...

  17. Not Your Run-of-the-Mill Art-Room Stools

    Science.gov (United States)

    Chrzanowski, Rose-Ann C.

    2010-01-01

    An art room should be a garden of visual stimulation, born of creativity, inquiry, critical thinking and intellectual conversation--and a little collaboration is not a bad thing either! When the author unpacked the new stools for her art room at the high school, she envisioned something more beautiful than the brown masonite circles that…

  18. Bristol scale stool form. A still valid help in medical practice and clinical research.

    Science.gov (United States)

    Riegler, G; Esposito, I

    2001-12-01

    The collection of clinical data concerning bowel habit is always empirical. A more extended use of visual descriptive stool form scales could contribute to a clearer and more standardized reporting of data about bowel function. This could be helpful for both clinical practice and research purposes.

  19. Differential clinical features and stool findings in shigellosis and amoebic dysentery

    NARCIS (Netherlands)

    Speelman, P.; McGlaughlin, R.; Kabir, I.; Butler, T.

    1987-01-01

    To obtain information that could assist the clinician to differentiate between shigellosis and amoebic dysentery, we compared clinical features and stool findings in 58 adult male patients in Bangladesh. Mean values indicated that patients with invasive amoebiasis were older and had a longer

  20. An uncommon case of late-onset congenital diaphragmatic hernia with bloody stool

    Directory of Open Access Journals (Sweden)

    Keisuke Jimbo

    2016-10-01

    Full Text Available Late-onset congenital diaphragmatic hernia (CDH is an uncommon subset of CDH and distinct from neonatal CDH with respect to presenting symptoms, diagnosis, management, and prognosis. In particular, CDH diagnosed after 30 days of age (late-onset CDH is uncommon and has an atypical presentation and a more favorable prognosis. In the present report, an infantile late-onset CDH case that presented with bloody stool and had a severe clinical course is described. In previous reports, no late-onset CDH case developed bloody stool. After diagnosis with image inspections, emergency surgery was performed. At operation, via the hernia orifice, the jejunum was seen to have prolapsed into the thoracic cavity with focally significant intestinal and mesenteric congestion, but no intestinal necrosis. In general, other disorders such as intussusception may be considered in the differential diagnosis of acute abdomen with bloody stool in patients of this age. However, in such late-onset CDH cases, immediate differentiation from other causes of acute abdomen that present with bloody stool is life-saving.

  1. New ultrasonographic evaluation of stool and/or gas distribution for treatment of chronic constipation.

    Science.gov (United States)

    Manabe, Noriaki; Kamada, Tomoari; Hata, Jiro; Haruma, Ken

    2018-03-01

    The first aim of this study was to develop a new ultrasonographic method (US) to evaluate stool and/or gas distribution. The second aim was to apply this method to compare stool and/or gas distribution between healthy subjects and patients with chronic constipation and evaluate whether US parameters could be an alternative to the colonic transit time (CTT). We enrolled seven healthy volunteers (four men, three women; mean age 29.3 ± 5.2 years) who underwent US and computed tomography (CT) on the same day to evaluate the reproducibility of US results. We then enrolled 268 patients with chronic constipation (94 men, 174 women; mean age 63.3 ± 4.2 years) and 66 age- and sex-matched healthy subjects (controls). The transverse diameters of four segments of the colon [ascending (AC), transverse (TC), descending (DC), and sigmoid (SC)] and the rectum (R) were measured, and their stool and/or gas distribution was evaluated using the constipation index (CI) [AC + TC + DC + SC + R/5] and left/right (L/R) distribution [(DC + SC)/(AC + TC)]. The CTT was assessed using radiopaque markers. All healthy subjects underwent US and CT successfully, with a sufficiently high reproducibility coefficient for this method and significant correlation between the US and CT parameters. The stool and/or gas distribution evaluated by US showed a significant difference in one of the US parameters between healthy subjects and patients, and the CI was an indirect indicator for the CTT. These findings may assist physicians evaluate stool and/or gas distribution of patients with chronic constipation, which is an indirect indicator for CTT.

  2. Development of single step RT-PCR for detection of Kyasanur forest disease virus from clinical samples

    Directory of Open Access Journals (Sweden)

    Gouri Chaubal

    2018-02-01

    Discussion and conclusion: The previously published sensitive real time RT-PCR assay requires higher cost in terms of reagents and machine setup and technical expertise has been the primary reason for development of this assay. A single step RT-PCR is relatively easy to perform and more cost effective than real time RT-PCR in smaller setups in the absence of Biosafety Level-3 facility. This study reports the development and optimization of single step RT-PCR assay which is more sensitive and less time-consuming than nested RT-PCR and cost effective for rapid diagnosis of KFD viral RNA.

  3. Cholera diagnosis in human stool and detection in water: protocol for a systematic review of available technologies.

    Science.gov (United States)

    Diaconu, Karin; Falconer, Jennifer; O'May, Fiona; Jimenez, Miguel; Matragrano, Joe; Njanpop-Lafourcade, Betty; Ager, Alastair

    2018-02-20

    Cholera is a highly infectious diarrheal disease spread via fecal contamination of water and food sources; it is endemic in parts of Africa and Asia and recent outbreaks have been reported in Haiti, the Zambia and Democratic Republic of the Congo. If left untreated, the disease can be fatal in less than 24 h and result in case fatality ratios of 30-50%. Cholera disproportionately affects those living in areas with poor access to water and sanitation: the long-term public health response is focused on improving water and hygiene facilities and access. Short-term measures for infection prevention and control, and disease characterization and surveillance, are impaired by diagnostic delays: culture methods are slow and rely on the availability of infrastructure and specialist equipment. Rapid diagnostic tests have shown promise under field conditions and further innovations in this area have been proposed. This paper is the protocol for a systematic review focused on identifying current technologies and methods used for cholera diagnosis in stool, and detection in water. We will synthesize and appraise information on product technical specifications, accuracy and design features in order to inform infection prevention and control and innovation development. Embase, MEDLINE, CINAHL, Proquest, IndMed and the WHO and Campbell libraries will be searched. We will include studies reporting on field evaluations, including within-study comparisons against a reference standard, and laboratory evaluations reporting on product validation against field stool or water samples. We will extract data according to protocol and attempt meta-analyses if appropriate given data availability and quality. The systematic review builds on a previous scoping review in this field and expands upon this by synthesising data on both product technical characteristics and design features. The review will be of particular value to stakeholders engaged in diagnostic procurement and manufacturers

  4. Cholera diagnosis in human stool and detection in water: protocol for a systematic review of available technologies

    Directory of Open Access Journals (Sweden)

    Karin Diaconu

    2018-02-01

    Full Text Available Abstract Background Cholera is a highly infectious diarrheal disease spread via fecal contamination of water and food sources; it is endemic in parts of Africa and Asia and recent outbreaks have been reported in Haiti, the Zambia and Democratic Republic of the Congo. If left untreated, the disease can be fatal in less than 24 h and result in case fatality ratios of 30–50%. Cholera disproportionately affects those living in areas with poor access to water and sanitation: the long-term public health response is focused on improving water and hygiene facilities and access. Short-term measures for infection prevention and control, and disease characterization and surveillance, are impaired by diagnostic delays: culture methods are slow and rely on the availability of infrastructure and specialist equipment. Rapid diagnostic tests have shown promise under field conditions and further innovations in this area have been proposed. Methods This paper is the protocol for a systematic review focused on identifying current technologies and methods used for cholera diagnosis in stool, and detection in water. We will synthesize and appraise information on product technical specifications, accuracy and design features in order to inform infection prevention and control and innovation development. Embase, MEDLINE, CINAHL, Proquest, IndMed and the WHO and Campbell libraries will be searched. We will include studies reporting on field evaluations, including within-study comparisons against a reference standard, and laboratory evaluations reporting on product validation against field stool or water samples. We will extract data according to protocol and attempt meta-analyses if appropriate given data availability and quality. Discussion The systematic review builds on a previous scoping review in this field and expands upon this by synthesising data on both product technical characteristics and design features. The review will be of particular value to

  5. Geographical Variation in Antibiotic-Resistant Escherichia coli Isolates from Stool, Cow-Dung and Drinking Water

    Science.gov (United States)

    Sahoo, Krushna Chandra; Tamhankar, Ashok J.; Sahoo, Soumyakanta; Sahu, Priyadarshi Soumyaranjan; Klintz, Senia Rosales; Lundborg, Cecilia Stålsby

    2012-01-01

    Little information is available on relationships between the biophysical environment and antibiotic resistance. This study was conducted to investigate the antibiotic resistance pattern of Escherichia coli isolated from child stool samples, cow-dung and drinking water from the non-coastal (230 households) and coastal (187 households) regions of Odisha, India. Susceptibility testing of E. coli isolates (n = 696) to the following antibiotics: tetracycline, ampicillin/sulbactam, cefuroxime, cefotaxime, cefixime, cotrimoxazole, amikacin, ciprofloxacin, norfloxacin and nalidixic acid was performed by the disk diffusion method. Ciprofloxacin minimum inhibitory concentration (MIC) values were determined for ciprofloxacin-resistant isolates (n = 83). Resistance to at least one antibiotic was detected in 90% or more of the E. coli isolates. Ciprofloxacin MIC values ranged from 8 to 32 µg/mL. The odds ratio (OR) of resistance in E. coli isolates from children’s stool (OR = 3.1, 95% CI 1.18–8.01), cow-dung (OR = 3.6, 95% CI 1.59–8.03, P = 0.002) and drinking water (OR = 3.8, 95% CI 1.00–14.44, P = 0.049) were higher in non-coastal compared to coastal region. Similarly, the co-resistance in cow-dung (OR = 2.5, 95% CI 1.39–4.37, P = 0.002) and drinking water (OR = 3.2, 95% CI 1.36–7.41, P = 0.008) as well as the multi-resistance in cow-dung (OR = 2.2, 95% CI 1.12–4.34, P = 0.022) and drinking water (OR = 2.7, 95% CI 1.06–7.07, P = 0.036) were also higher in the non-coastal compared to the coastal region. PMID:22690160

  6. Improved methodology for identification of protists and microalgae from plankton samples preserved in Lugol's iodine solution: combining microscopic analysis with single-cell PCR.

    Science.gov (United States)

    Auinger, Barbara M; Pfandl, Karin; Boenigk, Jens

    2008-04-01

    Here we introduce a method for quantitative analysis of planktonic protists and microalgae from preserved field samples combining morphological and small-subunit (SSU) rRNA gene sequence analysis. We linked a microscopic screening with PCR of single cells using field samples preserved with Lugol's iodine solution. Cells possessing a rigid cell wall were incubated with Viscozyme and subsequently with proteinase K for cell disruption; this was unnecessary for fragile cells. The addition of sodium thiosulfate to the PCR tube considerably decreased the inhibiting effect of the fixative (iodine) on the PCR and thus allowed for successful single-cell PCR even of long DNA fragments (up to as many as 3,000 base pairs). We further applied the protocol to investigate the dominant SSU rRNA genotypes in distinct flagellate morphospecies originating from different samples. We hypothesized that despite the morphological similarity, protist morphospecies in different habitats or sampled during different seasons are represented by different genotypes. Our results indicate species-specific differences: the two species Ochromonas sp. and Dinobryon divergens were represented by several different genotypes each, and for the latter species, the dominating genotype differed with habitat. In contrast, Dinobryon pediforme, Dinobryon bavaricum, and Synura sphagnicola were exclusively represented by a single genotype each, and the respective genotype was the same in different samples. In summary, our results highlight the significance of molecular variation within protist morphospecies.

  7. Improved Methodology for Identification of Protists and Microalgae from Plankton Samples Preserved in Lugol's Iodine Solution: Combining Microscopic Analysis with Single-Cell PCR▿ †

    Science.gov (United States)

    Auinger, Barbara M.; Pfandl, Karin; Boenigk, Jens

    2008-01-01

    Here we introduce a method for quantitative analysis of planktonic protists and microalgae from preserved field samples combining morphological and small-subunit (SSU) rRNA gene sequence analysis. We linked a microscopic screening with PCR of single cells using field samples preserved with Lugol's iodine solution. Cells possessing a rigid cell wall were incubated with Viscozyme and subsequently with proteinase K for cell disruption; this was unnecessary for fragile cells. The addition of sodium thiosulfate to the PCR tube considerably decreased the inhibiting effect of the fixative (iodine) on the PCR and thus allowed for successful single-cell PCR even of long DNA fragments (up to as many as 3,000 base pairs). We further applied the protocol to investigate the dominant SSU rRNA genotypes in distinct flagellate morphospecies originating from different samples. We hypothesized that despite the morphological similarity, protist morphospecies in different habitats or sampled during different seasons are represented by different genotypes. Our results indicate species-specific differences: the two species Ochromonas sp. and Dinobryon divergens were represented by several different genotypes each, and for the latter species, the dominating genotype differed with habitat. In contrast, Dinobryon pediforme, Dinobryon bavaricum, and Synura sphagnicola were exclusively represented by a single genotype each, and the respective genotype was the same in different samples. In summary, our results highlight the significance of molecular variation within protist morphospecies. PMID:18296536

  8. Hot topic: Bovine milk samples yielding negative or nonspecific results in bacterial culturing--the possible role of PCR-single strand conformation polymorphism in mastitis diagnosis.

    Science.gov (United States)

    Schwaiger, K; Wimmer, M; Huber-Schlenstedt, R; Fehlings, K; Hölzel, C S; Bauer, J

    2012-01-01

    A large proportion of mastitis milk samples yield negative or nonspecific results (i.e., no mastitis pathogen can be identified) in bacterial culturing. Therefore, the culture-independent PCR-single strand conformation polymorphism method was applied to the investigation of bovine mastitis milk samples. In addition to the known mastitis pathogens, the method was suitable for the detection of fastidious bacteria such as Mycoplasma spp., which are often missed by conventional culturing methods. The detection of Helcococcus ovis in 4 samples might indicate an involvement of this species in pathogenesis of bovine mastitis. In conclusion, PCR-single-strand conformation polymorphism is a promising tool for gaining new insights into the bacteriological etiology of mastitis. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. [Detection of Coxiella burnetii in dairy cattle bulk tank milk and single tank milk samples by confirmatory testing].

    Science.gov (United States)

    Hilbert, Angela; Andres, Tatjana; Werner, Ralf; Wehr, Roswitha; Fröhlich, Andreas; Conraths, Franz J; Henning, Klaus

    2015-01-01

    Q fever is a worldwide zoonotic disease caused by the pathogen Coxiella (C.) burnetii. A wide range of animal species is susceptible to this intracellular bacterium with great importance in ruminants. Human infections occur mainly by airborne transmission. C burnetii was detected in animal products such as raw milk, raw-milk cheese and butter prepared from raw milk as well as in the meat of infected animals. In cattle milk, the pathogen was detected up to 13 months after calving. The risk of human foodborne C. Burnetii infection is still considered to be low, but cannot be completely ruled out and remains under discussion. The aim of this study was to compare different laboratory diagnostic methods for C. burnetii in milk sample. The bulk tank and individual milk samples were sent and studied at the National Reference Laboratory for Q-fever in the context of confirmatory laboratory testing after clinical suspicion or retesting of previously antibody detection was in the analysis of 888 individual milk samples a match of 93.3% (Cohen-kappa). A total of 173 bulk milk samples and 2,807 individual milk samples from bovine herds for the presence of C. burnetii DNA and antibodies were tested against the pathogen. The pathogen was detected in 62.5% of the bulk milk samples and up to 60% in individual milk samples. The highest proportion of positive bulk milks was determined as 68.3% in 2012. In individual milk samples, the highest proportion of seropositive samples was 62.2%.

  10. Sample preparation for semivolatile organics analysis of Hanford single-shell tank waste with high nitrate/nitrite and water content

    International Nuclear Information System (INIS)

    Stromatt, R.W.; Hoppe, E.W.; Steele, M.J.

    1993-11-01

    This report describes research work carried out to solve sample preparation problems associated with applying gas chromatography with mass spectrometric detection (GC/MS) to the analysis of single shell tank (SST) samples from Hanford for semivolatile organic compounds. Poor performance was found when applying the procedures based on the U.S. Environmental Protection Agency (EPA), Contract Laboratory Program, Statement of Work (CLP SOW). Analysis work was carried out on simulated drainable liquid modeled after the SST core samples which had evidenced analysis problems. Some work was also conducted on true SST samples. It was found that the pH range was too broad in the original procedure. It was also found that by decreasing the amount of methanol used in the extraction process, problems associated with the formation of an azeotrope phase could be avoided. The authors suggest a new procedure, whose eventual application to a wide array of SST samples will lend itself to better quality control limits

  11. Evidence of heterozygosity and recombinant alleles in single cysts of Giardia duodenalis

    OpenAIRE

    Aguiar, Juliana Martins; Silva, Sheila Oliveira; Santos, Valdir Azevedo dos; Taniwaki, Sueli Akemi; Oliveira, Tricia Maria Ferreira de Sousa; Ferreira, Helena Lage; Keid, Lara Borges; Gregori, Fábio; Soares, Rodrigo Martins

    2016-01-01

    Abstract Giardia duodenalis is divided into eight assemblages (named A to H). Isolates of assemblage A are divided into four sub-assemblages (AI, AII, AIII and AIV). While isolates of sub-assemblage AII are almost exclusively detected in human hosts, isolates of assemblage B are encountered in a multitude of animal hosts and humans. Here, we isolated single cysts of G. duodenalis from a human stool sample and found that one of them had overlaps of assemblage AII and B alleles and an unexpecte...

  12. Performance evaluation of point-of-care test for detection of Cryptosporidium stool antigen in children and HIV infected adults.

    Science.gov (United States)

    Shimelis, Techalew; Tadesse, Endale

    2014-05-16

    Gastro-enteritis is associated with significant morbidity and mortality in patients with HIV/AIDS and children, and Cryptosporidium is the most important parasite implicated. To date, several commercial companies have developed simple and rapid point-of-care tests for the detection of Cryptosporidium infection; however, information is scarce regarding their diagnostic significance in Ethiopia. This study aimed at evaluating the performance of a rapid diagnostic test (RDT) for the detection of Cryptosporidium stool antigen. A hospital-based cross-sectional study was conducted in Hawassa University Hospital, southern Ethiopia from May to November 2013. Faecal samples were collected from a total of 100 children and 250 HIV infected individuals with diarrhea or CD4 T-cell count lower than 200 cells/μl. Specimens were processed using direct, formol-ether concentration and modified Ziehl-Neelsen techniques for diagnosis of Cryptosporidium and other parasites. One hundred faecal samples (50 positives for Cryptosporidium, 35 positives for other parasites and 15 negatives for any intestinal parasites) were tested using the CoproStrip™Cryptosporidium kit (Savyon Diagnostics Ltd, Israel). Test parameters were calculated using microscopy of the modified Ziehl-Neelsen stained stool smear as reference method. The performance of the RDT was first compared to routine microscopic analysis (examination ≤10 min). The CoproStrip™Cryptosporidium RDT correctly detected 31 of 42 positive samples and 49 of 50 negative samples (i.e., 11 false negatives and 1 false positive). Sensitivity, specificity, PPV, NPV and accuracy were calculated to be 74, 98, 97, 84 and 88%, respectively. Upon thorough microscopic analysis (examination >10 min), 8 more samples with very low oocyst density were found. However, these were missed by the kit and lower the sensitivity and NPV to 62 and 72%, respectively. No cross-reactivity was observed with any of the helminthic or other protozoan parasites

  13. Single-molecule X-ray free-electron laser imaging : Interconnecting sample orientation with explosion data

    OpenAIRE

    Östlin, Christofer

    2014-01-01

    X-ray crystallography has been around for 100 years and remains the preferred technique for solving molecular structures today. However, its reliance on the production of sufficiently large crystals is limiting, considering that crystallization cannot be achieved for a vast range of biomolecules. A promising way of circumventing this problem is the method of serial femtosecond imaging of single-molecules or nanocrystals utilizing an X-ray free-electron laser. In such an approach, X-ray pulses...

  14. A novel method for single sample multi-axial nanoindentation of hydrated heterogeneous tissues based on testing great white shark jaws.

    Directory of Open Access Journals (Sweden)

    Toni L Ferrara

    Full Text Available Nanomechanical testing methods that are suitable for a range of hydrated tissues are crucial for understanding biological systems. Nanoindentation of tissues can provide valuable insights into biology, tissue engineering and biomimetic design. However, testing hydrated biological samples still remains a significant challenge. Shark jaw cartilage is an ideal substrate for developing a method to test hydrated tissues because it is a unique heterogeneous composite of both mineralized (hard and non-mineralized (soft layers and possesses a jaw geometry that is challenging to test mechanically. The aim of this study is to develop a novel method for obtaining multidirectional nanomechanical properties for both layers of jaw cartilage from a single sample, taken from the great white shark (Carcharodon carcharias. A method for obtaining multidirectional data from a single sample is necessary for examining tissue mechanics in this shark because it is a protected species and hence samples may be difficult to obtain. Results show that this method maintains hydration of samples that would otherwise rapidly dehydrate. Our study is the first analysis of nanomechanical properties of great white shark jaw cartilage. Variation in nanomechanical properties were detected in different orthogonal directions for both layers of jaw cartilage in this species. The data further suggest that the mineralized layer of shark jaw cartilage is less stiff than previously posited. Our method allows multidirectional nanomechanical properties to be obtained from a single, small, hydrated heterogeneous sample. Our technique is therefore suitable for use when specimens are rare, valuable or limited in quantity, such as samples obtained from endangered species or pathological tissues. We also outline a method for tip-to-optic calibration that facilitates nanoindentation of soft biological tissues. Our technique may help address the critical need for a nanomechanical testing method

  15. Towards diffractive imaging with single pulses of FEL radiation. Dynamics within irradiatied samples and their influence on the analysis of imaging data

    International Nuclear Information System (INIS)

    Wang, Fenglin

    2010-08-01

    3D single particle coherent diffraction imaging (CDI) of bioparticles (such as proteins, macromolecules and viruses) is one of the main possible applications of the new generation of light sources: free-electron lasers (FELs), which are now available at FLASH (Hamburg, Germany) and LCLS (Stanford, U.S.A.). The extremely bright and ultrashort FEL pulses potentially enable CDI to achieve high resolution down to subnanometer length scale. However, intense FEL pulses cause serious radiation damage in bioparticles, even during single shots, which may set the resolution limits for CDI with FELs. Currently, since the signal-to-noise ratio is very low for small biological particles, direct experimental study of radiation damage in the single particle imaging is fairly difficult. Single atomic (noble gas) clusters become good objects to reveal effects of radiation damage processes on CDI with FEL radiation. This thesis studies three aspects of the radiation damage problem, which are treated in three independent chapters: (1) Molecular Dynamics simulations to quantitively describe radiation damage processes within irradiated atomic clusters during single pulses; (2) reconstruction analysis of single-shot CDI diffraction patterns of atomic clusters, which may potentially help to understand the radiation damage occurring in biological samples; and (3) testing the effects of coating water layers in CDI, which is supposed to minimize the radiation damage in irradiated bioparticles. (orig.)

  16. Towards diffractive imaging with single pulses of FEL radiation. Dynamics within irradiatied samples and their influence on the analysis of imaging data

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Fenglin

    2010-08-15

    3D single particle coherent diffraction imaging (CDI) of bioparticles (such as proteins, macromolecules and viruses) is one of the main possible applications of the new generation of light sources: free-electron lasers (FELs), which are now available at FLASH (Hamburg, Germany) and LCLS (Stanford, U.S.A.). The extremely bright and ultrashort FEL pulses potentially enable CDI to achieve high resolution down to subnanometer length scale. However, intense FEL pulses cause serious radiation damage in bioparticles, even during single shots, which may set the resolution limits for CDI with FELs. Currently, since the signal-to-noise ratio is very low for small biological particles, direct experimental study of radiation damage in the single particle imaging is fairly difficult. Single atomic (noble gas) clusters become good objects to reveal effects of radiation damage processes on CDI with FEL radiation. This thesis studies three aspects of the radiation damage problem, which are treated in three independent chapters: (1) Molecular Dynamics simulations to quantitively describe radiation damage processes within irradiated atomic clusters during single pulses; (2) reconstruction analysis of single-shot CDI diffraction patterns of atomic clusters, which may potentially help to understand the radiation damage occurring in biological samples; and (3) testing the effects of coating water layers in CDI, which is supposed to minimize the radiation damage in irradiated bioparticles. (orig.)

  17. Determination of the thermal neutron absorption cross section for rock samples by a single measurement of the time decay constant

    International Nuclear Information System (INIS)

    Krynicka, E.

    1993-01-01

    A calibration method for the determination of the thermal neutron macroscopic mass absorption cross section for rock samples is presented. The standard deviation of the final results is discussed in detail. A big advantage of the presented method is that the calibration curves have been found using the results obtained for a variety of natural rock samples of different stratigraphies and lithologies measured by Czubek's methods. An important part of the paper is a through analysis of the standard deviation of the final result. (author). 13 refs, 11 figs, 5 tabs

  18. Optimal sample size for predicting viability of cabbage and radish seeds based on near infrared spectra of single seeds

    DEFF Research Database (Denmark)

    Shetty, Nisha; Min, Tai-Gi; Gislum, René

    2011-01-01

    The effects of the number of seeds in a training sample set on the ability to predict the viability of cabbage or radish seeds are presented and discussed. The supervised classification method extended canonical variates analysis (ECVA) was used to develop a classification model. Calibration sub...... and radish data. The misclassification rates at optimal sample size were 8%, 6% and 7% for cabbage and 3%, 3% and 2% for radish respectively for random method (averaged for 10 iterations), DUPLEX and CADEX algorithms. This was similar to the misclassification rate of 6% and 2% for cabbage and radish obtained...

  19. Quantitative Detection of Shiga Toxins Directly from Stool Specimens of Patients Associated with an Outbreak of Enterohemorrhagic Escherichia coli in Japan--Quantitative Shiga toxin detection from stool during EHEC outbreak.

    Science.gov (United States)

    Yamasaki, Eiki; Watahiki, Masanori; Isobe, Junko; Sata, Tetsutaro; Nair, G Balakrish; Kurazono, Hisao

    2015-10-27

    Detection of Shiga toxins (Stx) is important for accurate diagnosis of Enterohemorrhagic Escherichia coli infection. In this study, we quantitatively analyzed Stx protein in nine patients' stool during an outbreak that occurred in Japan. Highly sensitive immunoassay (bead enzyme-linked immunosorbent assay (bead-ELISA)) revealed that the concentrations of toxins in stool of patients ranged from 0.71 to 10.44 ng/mL for Stx1 and 2.75 to 51.61 ng/mL for Stx2. To our knowledge, this is the first report that reveals the range of Stx protein concentrations in human stools.

  20. Randomized controlled trial: Standard versus supplemental bowel preparation in patients with Bristol stool form 1 and 2.

    Science.gov (United States)

    Li, Yueyue; Jia, Xinyong; Liu, Baozhen; Qi, Yanmei; Zhang, Xiubin; Ji, Rui; Yu, Yanbo; Zuo, Xiuli; Li, Yanqing

    2017-01-01

    Bristol stool form 1 and 2 is an important predictor of inadequate bowel preparation. To evaluate the efficacy of supplemental preparation in bowel cleansing quality among patients with Bristol stool form 1 and 2, as well as the feasibility of tailored bowel preparation guided by Bristol stool form scale. Patients with Bristol stool form 1 and 2 from 3 Chinese tertiary hospitals randomly received either 2 L PEG-ELP (group A) or 10 mg bisacodyl plus 2 L PEG-ELP (group B); patients with Bristol stool form 3 to 7 received 2 L PEG-ELP (group C) for bowel preparation. The primary endpoint is the rate of adequate bowel reparation for the whole colon. The adequate bowel preparation rate for separate colon segments, the polyp detection rate (PDR), tolerability, acceptability, sleeping quality and compliance were evaluated as secondary endpoints. 700 patients were randomized. In per-protocol analysis, patients in group B attained significantly higher successful preparation rate than group A (88.7% vs. 61.2%, pBristol stool form 1 and 2. Bristol stool form scale may be an easy and efficient guide for tailored bowel preparation before colonoscopy.

  1. Prediction of the systemic exposure to oral 9-amino-20(S)-camptothecin using single-sample analysis

    NARCIS (Netherlands)

    Sparreboom, A.; de Jonge, M. J.; Punt, C. J.; Loos, W. J.; Nooter, K.; Stoter, G.; Porro, M. G.; Verweij, J.

    1999-01-01

    The purpose of this study was to develop and validate limited-sampling strategies for prediction of the area under the plasma-concentration time curves (AUCs) of the lactone and total (i. e., lactone plus carboxylate) forms of the novel topoisomerase-I inhibitor 9-amino-20(S)-camptothecin (9-AC).

  2. Prediction of the systemic exposure to oral 9-amino-20(S)-camptothecin using single-sample analysis

    NARCIS (Netherlands)

    A. Sparreboom (Alex); M.J.A. de Jonge (Maja); C.J.A. Punt (Cornelis); W.J. Loos (Walter); K. Nooter (Kees); G. Stoter (Gerrit); M.G. Porro; J. Verweij (Jaap)

    1999-01-01

    textabstractThe purpose of this study was to develop and validate limited-sampling strategies for prediction of the area under the plasma-concentration time curves (AUCs) of the lactone and total (i. e., lactone plus carboxylate) forms of the novel topoisomerase-I

  3. Single Image Super-resolution Using Gaussian Process Regression with Dictionary-based Sampling and Student-t Likelihood.

    Science.gov (United States)

    Wang, Haijun; Gao, Xinbo; Zhang, Kaibing; Li, Jie

    2017-05-03

    Gaussian process regression (GPR) is an effective statistical learning method for modeling non-linear mapping from an observed space to an expected latent space. When applying it to example learning-based super-resolution (SR), two outstanding issues remain. One is its high computational complexity restricts SR application when a large dataset is available for learning task. The other is that the commonly used Gaussian likelihood in GPR is incompatible with the true observation model for SR reconstruction. To alleviate the above two issues, we propose a GPR-based SR method by using dictionary-based sampling and student-t likelihood, called DSGPR. Considering that dictionary atoms effectively span the original training sample space, we adopt a dictionary-based sampling strategy by combining all the neighborhood samples of each atom into a compact representative training subset so as to reduce the computational complexity. Based on statistical tests, we statistically validate that Studentt likelihood is more suitable to build the observation model for SR problem. Extensive experimental results show that the proposed method outperforms other competitors and produces more pleasing details in texture regions.

  4. Mites in dust samples from mattress surfaces from single beds or cribs in the south Brazilian city of Londrina.

    Science.gov (United States)

    da Silva, Dagoberto Ribeiro; Binotti, Raquel Soares; da Silva, Cleide Moreira; de Oliveira, Celso Henrique; Condino-Neto, Antônio; de Capitani, Eduardo Mello

    2005-03-01

    The aim of this study was to investigate the mite fauna in mattresses dust samples from cribs or beds in the south Brazilian city of Londrina, State of Parana. A total of 133 dust samples from upper and lower mattress surfaces, and bed frames were aspirated once from 38 dwellings (18 cribs and 21 beds), and one day nursery (six cribs). A total of 758 mite bodies were counted in slides: 233 (30.7%) from cribs and 525 (69.3%) from beds (pdust mites--mainly Dermatophagoides pteronyssinus, represented 72% and 84% of total mite count in crib and bed dust samples, respectively. The mean HDM body concentration in crib or bed slides were, respectively, 289.9+/-136.7 and 875.0+/-183.6 mites/g. Statistical analysis showed a significantly higher mite bodies count on lower mattress surface compared with upper surface in bed samples only (p=0.025). Data herein show that cribs like mattress have sufficient mite bodies to cause sensitization to humans. The use of mattress covers for cribs and beds should be encouraged in order to avoid allergens exposure. Copyright (c) 2005 Blackwell Munksgaard

  5. Single-shot all-optical sampling oscilloscope using a polarization-maintaining resonator for pulse replication

    Czech Academy of Sciences Publication Activity Database

    Komanec, M.; Honzátko, Pavel; Zvánovec, S.

    2010-01-01

    Roč. 52, č. 11 (2010), s. 2452-2456 ISSN 0895-2477 R&D Projects: GA AV ČR 1ET300670502 Institutional research plan: CEZ:AV0Z20670512 Keywords : optical sampling oscilloscope * four-wave mixing * fiber pulse replicator * highly nonlinear fiber Subject RIV: BH - Optics, Masers, Lasers Impact factor: 0.656, year: 2010

  6. Simple methodology to directly genotype Trypanosoma cruzi discrete typing units in single and mixed infections from human blood samples.

    Science.gov (United States)

    Bontempi, Iván A; Bizai, María L; Ortiz, Sylvia; Manattini, Silvia; Fabbro, Diana; Solari, Aldo; Diez, Cristina

    2016-09-01

    Different DNA markers to genotype Trypanosoma cruzi are now available. However, due to the low quantity of parasites present in biological samples, DNA markers with high copy number like kinetoplast minicircles are needed. The aim of this study was to complete a DNA assay called minicircle lineage specific-PCR (MLS-PCR) previously developed to genotype the T. cruzi DTUs TcV and TcVI, in order to genotype DTUs TcI and TcII and to improve TcVI detection. We screened kinetoplast minicircle hypervariable sequences from cloned PCR products from reference strains belonging to the mentioned DTUs using specific kDNA probes. With the four highly specific sequences selected, we designed primers to be used in the MLS-PCR to directly genotype T. cruzi from biological samples. High specificity and sensitivity were obtained when we evaluated the new approach for TcI, TcII, TcV and TcVI genotyping in twenty two T. cruzi reference strains. Afterward, we compared it with hybridization tests using specific kDNA probes in 32 blood samples from chronic chagasic patients from North Eastern Argentina. With both tests we were able to genotype 94% of the samples and the concordance between them was very good (kappa=0.855). The most frequent T. cruzi DTUs detected were TcV and TcVI, followed by TcII and much lower TcI. A unique T. cruzi DTU was detected in 18 samples meantime more than one in the remaining; being TcV and TcVI the most frequent association. A high percentage of mixed detections were obtained with both assays and its impact was discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Three-step stool examination for cryptosporidiosis in 10 homosexual men with protracted watery diarrhea.

    Science.gov (United States)

    Ma, P; Soave, R

    1983-05-01

    Cryptosporidiosis, a zoonosis caused by Cryptosporidium species, is a newly recognized coccidial protozoan infection causing severe protracted watery diarrhea in humans. In August 1981, the first case of cryptosporidiosis in a homosexual man with acquired immune deficiency syndrome (AIDS) was reported; diagnosis was determined by intestinal biopsy. It is necessary to adopt a simple laboratory diagnostic procedure to screen large numbers of suspected cases. A three-step stool examination was developed to demonstrate Cryptosporidium oocysts and the diagnostic and infective stages of the infection in 10 homosexual men with AIDS. This is a less invasive, less costly, and more sensitive test than intestinal biopsy and has been designed to prevent confusion caused by yeast cells that are frequently present in stool, leading to a false diagnosis. The examination consists of preliminary differential determination by iodine wet mount, definitive identification by modified Kinyoun acid-fast staining, and a more effective method of concentrating oocysts, by Sheather's sugar cover-slip flotation method.

  8. Water absorption kinetics in different wettability conditions studied at pore and sample scales in porous media by NMR with portable single-sided and laboratory imaging devices

    Science.gov (United States)

    Bortolotti, V.; Camaiti, M.; Casieri, C.; De Luca, F.; Fantazzini, P.; Terenzi, C.

    2006-08-01

    NMR relaxation time distributions of water 1H obtained by a portable single-sided surface device have been compared with MRI internal images obtained with a laboratory imaging apparatus on the same biocalcarenite (Lecce Stone) samples during capillary water uptake. The aim of this work was to check the ability of NMR methods to quantitatively follow the absorption phenomenon under different wettability conditions of the internal pore surfaces. Stone wettability changes were obtained by capillary absorption of a chloroform solution of Paraloid PB72, a hydrophobic acrylic resin frequently used to protect monuments and buildings, through one face of each sample. Both relaxation and imaging data have been found in good quantitative agreement each other and with masses of water determined by weighing the samples. In particular the Washburn model of water capillary rise applied to the imaging data allowed us to quantify the sorptivity in both treated and untreated samples. Combining relaxation and imaging data, a synergetic improvement of our understanding of the water absorption kinetics at both pore and sample scales is obtained. Since relaxation data have been taken over the course of time without interrupting the absorption process, simply by keeping the portable device on the surface opposite to the absorption, the results show that the single-sided NMR technique is a powerful tool for in situ evaluation of water-repellent treatments frequently used for consolidation and/or protection of stone artifacts.

  9. The effect of inclined step stool on the quality of chest compression during in-hospital cardiopulmonary resuscitation.

    Science.gov (United States)

    Yun, Seong-Woo; Lee, Byung Kook; Jeung, Kyung Woon; Park, Sang Wook; Choi, Sung Soo; Lee, Chang-Hee; Ryu, So-Yeon

    2014-08-01

    A step stool is an ordinary device to improve the quality of chest compression (CC) during in-hospital cardiopulmonary resuscitation (CPR). We investigated the effect of an inclined step stool on the quality of CC during CPR on a hospital bed. We conducted a randomized crossover study of simulation using a manikin. Two different methods of CC were performed and compared: CC using a flat stool and CC using an inclined (20°) stool. Each session of CC was performed for 2 minutes using a metronome at a rate of 110 beats per minute. The primary outcome was the depth of CC. The adequate CC rate, duty cycle, rate of incomplete recoil, and the angle between the arm of the participants and the bed were also measured. The median value of the mean depth of CC was 50.5 mm (45.0-57.0 mm) in the flat stool group and 54.5 mm (47.0-58.3 mm) in the inclined stool group (P = .014). The adequate CC rate was significantly higher in the inclined stool group (84.2% [37.6%-99.1%] vs 57.0% [15.2%-95.0%]; P = .016). The duty cycle and the rate of incomplete recoil were comparable between the 2 groups. The angles between the arm of the participants and the bed were more vertical in the inclined stool group (84.0° ± 5.2° vs 81.0° ± 4.8°; P = .014). Using an inclined stool resulted in an improvement in the depth of CC and the adequate CC rate without increasing the rate of incomplete chest recoil. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens.

    OpenAIRE

    Garcia, L S; Bruckner, D A; Brewer, T C; Shimizu, R Y

    1983-01-01

    Due to increasing numbers of patients with documented infections with Cryptosporidium and other coccidia, it is important for the physician and clinical laboratory to be aware of the appropriate diagnostic techniques necessary for organism recovery and identification. Although Cryptosporidium is found in the gastrointestinal tract, tissue biopsies may be insufficient for organism recovery; the examination of stool specimens is a noninvasive procedure and will provide better overall opportunit...

  11. Multicenter clinical evaluation of the portrait toxigenic C. difficile assay for detection of toxigenic Clostridium difficile strains in clinical stool specimens.

    Science.gov (United States)

    Buchan, Blake W; Mackey, Tami-Lea A; Daly, Judy A; Alger, Garrison; Denys, Gerald A; Peterson, Lance R; Kehl, Sue C; Ledeboer, Nathan A

    2012-12-01

    We compared the Portrait Toxigenic C. difficile Assay, a new semiautomated sample-to-result molecular test, to a toxigenic bacterial culture/cell cytotoxin neutralization assay (TBC/CCNA) for the detection of toxigenic Clostridium difficile in 549 stool specimens. Stool specimens were also tested by one of three alternative FDA-cleared molecular tests for toxigenic C. difficile (Xpert C. difficile, Illumigene C. difficile, or GeneOhm Cdiff). The sensitivities and specificities of the molecular tests compared to TBC/CCNA were as follows: 98.2% and 92.8% for the Portrait assay, 100% and 91.7% for the Xpert assay, 93.3% and 95.1% for the Illumigene assay, and 97.4% and 98.5% for the GeneOhm assay, respectively. The majority of Portrait false-positive results (20/31; 64.5%) were also positive for C. difficile by an alternative molecular test, suggesting an increased sensitivity compared to the culture-based "gold standard" method. The Portrait test detected an assay input of 30 CFU in 100% of spiked samples and detected an input of 10 CFU in 96.7% of samples tested.

  12. Determination of the optional time for taking blood samples by single intravenous injection of 3H-leucine

    International Nuclear Information System (INIS)

    Meng Delian; Yao Junhu; Lu Jinyin; Wu Xiaobin; Liu Jun

    2003-01-01

    Twenty four young hens (1.5 kg of body weight, BW) were randomly divided into 4 groups. Every group was diet free (FAS) or force-fed a nitrogen-free diet (NFD) or the diet with 20% crude protein in which soybean meal or cotton seed meal was the sole nitrogen source (30 g DM/kg BW). 30 μCi 3 H-Leu/kg BW was intravenously injected into all birds just after force-fed or on fasting. Venous blood samples were taken at 5, 30 min, 4,24,36 and 48h after injection. The excreta during the whole period of 48h after injection was collected. Special radioactivities of nonprotein plasma at every time point and excreta were measured. The optional time of taking blood samples was 20-24 hours after injected 3 H-Leu

  13. Forensic Sampling and Analysis from a Single Substrate: Surface-Enhanced Raman Spectroscopy Followed by Paper Spray Mass Spectrometry.

    Science.gov (United States)

    Fedick, Patrick W; Bills, Brandon J; Manicke, Nicholas E; Cooks, R Graham

    2017-10-17

    Sample preparation is the most common bottleneck in the analysis and processing of forensic evidence. Time-consuming steps in many forensic tests involve complex separations, such as liquid and gas chromatography or various types of extraction techniques, typically coupled with mass spectrometry (e.g., LC-MS). Ambient ionization ameliorates these slow steps by reducing or even eliminating sample preparation. While some ambient ionization techniques have been adopted by the forensic community, there is significant resistance to discarding chromatography as most forensic analyses require both an identification and a confirmation technique. Here, we describe the use of a paper substrate, the surface of which has been inkjet printed with silver nanoparticles, for surface enhanced Raman spectroscopy (SERS). The same substrate can also act as the paper substrate for paper spray mass spectrometry. The coupling of SERS and paper spray ionization creates a quick, forensically feasible combination.

  14. Stool Tests

    Science.gov (United States)

    ... be messy, so be sure to wear latex gloves and wash your hands and your child's hands ... sometimes give clues about certain illnesses, especially in newborns or very ill children. Viral cultures can take ...

  15. Stool Culture

    Science.gov (United States)

    ... illness by producing toxins . These bacteria may be cultured , but many of the tests used to detect ... been contaminated with pathogenic bacteria, such as undercooked meat or raw eggs, or the same food that ...

  16. Rapid Treponema pallidum clearance from blood and ulcer samples following single dose benzathine penicillin treatment of early syphilis.

    Science.gov (United States)

    Tipple, Craig; Jones, Rachael; McClure, Myra; Taylor, Graham

    2015-02-01

    Currently, the efficacy of syphilis treatment is measured with anti-lipid antibody tests. These can take months to indicate cure and, as a result, syphilis treatment trials require long periods of follow-up. The causative organism, Treponema pallidum (T. pallidum), is detectable in the infectious lesions of early syphilis using DNA amplification. Bacteraemia can likewise be identified, typically in more active disease. We hypothesise that bacterial clearance from blood and ulcers will predict early the standard serology-measured treatment response and have developed a qPCR assay that could monitor this clearance directly in patients with infectious syphilis. Patients with early syphilis were given an intramuscular dose of benzathine penicillin. To investigate the appropriate sampling timeframe samples of blood and ulcer exudate were collected intensively for T. pallidum DNA (tpp047 gene) and RNA (16S rRNA) quantification. Sampling ended when two consecutive PCRs were negative. Four males were recruited. The mean peak level of T. pallidum DNA was 1626 copies/ml whole blood and the mean clearance half-life was 5.7 hours (std. dev. 0.53). The mean peak of 16S rRNA was 8879 copies/ml whole blood with a clearance half-life of 3.9 hours (std. dev. 0.84). From an ulcer, pre-treatment, 67,400 T. pallidum DNA copies and 7.08 x 107 16S rRNA copies were detected per absorbance strip and the clearance half-lives were 3.2 and 4.1 hours, respectively. Overall, T. pallidum nucleic acids were not detected in any sample collected more than 56 hours (range 20-56) after treatment. All patients achieved serologic cure. In patients with active early syphilis, measuring T. pallidum levels in blood and ulcer exudate may be a useful measure of treatment success in therapeutic trials. These laboratory findings need confirmation on a larger scale and in patients receiving different therapies.

  17. A comparison of three fecal steroid metabolites for pregnancy detection used with single sampling in bighorn sheep (Ovis canadensis)

    Science.gov (United States)

    Schoenecker, K.A.; Lyda, R.O.; Kirkpatrick, J.

    2004-01-01

    We compared three fecal steroid metabolite assays for their usefulness in detecting pregnancy among free-ranging Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from Bighorn Canyon National Recreation Area, Wyoming and Montana (USA) and captive bighorn ewes at ZooMontana in Billings, Montana. Fecal samples were collected from 11 free-ranging, radio-collared bighorn ewes in late January–May 2001 and from 20 free-ranging, radio-collared ewes in late March to mid-May 2002. Free-ranging ewes were monitored the following spring to determine whether or not they lambed. In addition, two captive ewes were studied at Zoo-Montana. With three exceptions, free-ranging bighorn ewes that produced lambs had nonspecific progesterone metabolite (iPdG) levels of >1,800 ng/g feces and iPdG levels >7,000 ng/gm feces when samples were collected between early March and mid-May Samples collected earlier in the year were inconclusive. One false negative was suspected to be the result of sample collection error. Of the captive ewes, nonspecific pregnanediol-3α–glucuronide (PdG) and iPdG followed a predictable curve over the course of the 180-day pregnancies. We conclude that estrone conjugates are not useful in diagnosing pregnancy; however, fecal steroid analysis of PdG and iPdG can be used to accurately determine pregnancy and reproductive function in bighorn sheep. This holds great potential as a noninvasive technique for understanding the role of reproductive disease in wild bighorn sheep.

  18. Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens.

    Science.gov (United States)

    Garcia, L S; Bruckner, D A; Brewer, T C; Shimizu, R Y

    1983-07-01

    Due to increasing numbers of patients with documented infections with Cryptosporidium and other coccidia, it is important for the physician and clinical laboratory to be aware of the appropriate diagnostic techniques necessary for organism recovery and identification. Although Cryptosporidium is found in the gastrointestinal tract, tissue biopsies may be insufficient for organism recovery; the examination of stool specimens is a noninvasive procedure and will provide better overall opportunities for organism recovery. Human clinical specimens were examined from 45 patients with confirmed cryptosporidiosis or suspected of having the infection. Tissue biopsy sections, fecal wet preparations, and permanent stained smears were examined. Stool specimens were submitted in 10% Formalin, 2.5% potassium dichromate, and polyvinyl alcohol and were examined for oocysts by using 15 different methods: phase-contrast and light microscopy; Sheather's sugar flotation; Formalin concentration techniques; 10% potassium hydroxide; Giemsa; trichrome; periodic acid-Schiff; modified periodic acid-Schiff; silver methenamine; acridine orange; auramine-rhodamine; Kinyoun acid-fast; Ziehl-Neelsen carbolfuchsin; and a modified acid-fast procedure. Each technique or combination of techniques was assessed by organism quantitation, organism morphology, and ease of visual recognition. Based on these comparative studies, the modified Ziehl-Neelsen carbolfuchsin stain on 10% Formalin-preserved stool is recommended for the recovery and identification of Cryptosporidium.

  19. Detection of Colorectal Cancer by a Quantitative Fluorescence Determination of DNA Amplification in Stool

    Directory of Open Access Journals (Sweden)

    Daniele Calistri

    2004-09-01

    Full Text Available DNA amplification of exfoliated cells in stool repre sents an inexpensive and rapid test, but has only 50% to 60% sensitivity. A new quantitative method, calle( fluorescence long DNA, was developed and validate( in our laboratory on stool obtained from 86 patient., with primary colorectal cancer and from 62 health individuals. It consists of the amplification of stoo DNA with fluorescence primers and the quantification of the amplification using a standard curve. Results are arbitrarily expressed in nanograms. The potential of thi new method compared to the conventional approact was analyzed in a subgroup of 94 individuals (51 patients and 38 healthy volunteers. In the presen series, DNA amplification analysis showed a specific ity of 97% and a sensitivity of only 50%. Conversely fluorescence DNA evaluation, using the best cutoff o 25 ng, showed a sensitivity of about 76% and a spec ificity of 93%. Similar sensitivity was observed regard less of Dukes stage, tumor location, and size, thu., also permitting the detection of early-stage tumors The present study seems to indicate that quantitative fluorescence DNA determination in stool successfully identifies colorectal cancer patients with a sensitivity comparable, if not superior, to that of multiple gene analysis but at a lower cost and in a shorter time.

  20. Multifaceted empathy of healthy volunteers after single doses of MDMA: A pooled sample of placebo-controlled studies.

    Science.gov (United States)

    Kuypers, Kim Pc; Dolder, Patrick C; Ramaekers, Johannes G; Liechti, Matthias E

    2017-05-01

    Previous placebo-controlled experimental studies have shown that a single dose of MDMA can increase emotional empathy in the multifaceted empathy test (MET) without affecting cognitive empathy. Although sufficiently powered to detect main effects of MDMA, these studies were generally underpowered to also validly assess contributions of additional parameters, such as sex, drug use history, trait empathy and MDMA or oxytocin plasma concentrations. The present study examined the robustness of the MDMA effect on empathy and investigated the moderating role of these additional parameters. Participants ( n = 118) from six placebo-controlled within-subject studies and two laboratories were included in the present pooled analysis. Empathy (MET), MDMA and oxytocin plasma concentrations were assessed after oral administration of MDMA (single dose, 75 or 125 mg). Trait empathy was assessed using the interpersonal reactivity index. We confirmed that MDMA increased emotional empathy at both doses without affecting cognitive empathy. This MDMA-related increase in empathy was most pronounced during presentation of positive emotions as compared with negative emotions. MDMA-induced empathy enhancement was positively related to MDMA blood concentrations measured before the test, but independent of sex, drug use history and trait empathy. Oxytocin concentrations increased after MDMA administration but were not associated with behavioral effects. The MDMA effects on emotional empathy were stable across laboratories and doses. Sex did not play a moderating role in this effect, and oxytocin levels, trait empathy and drug use history were also unrelated. Acute drug exposure was of significant relevance in the MDMA-induced emotional empathy elevation.

  1. Efficient sampling of reversible cross-linking polymers: Self-assembly of single-chain polymeric nanoparticles

    Science.gov (United States)

    Oyarzún, Bernardo; Mognetti, Bortolo Matteo

    2018-03-01

    We present a new simulation technique to study systems of polymers functionalized by reactive sites that bind/unbind forming reversible linkages. Functionalized polymers feature self-assembly and responsive properties that are unmatched by the systems lacking selective interactions. The scales at which the functional properties of these materials emerge are difficult to model, especially in the reversible regime where such properties result from many binding/unbinding events. This difficulty is related to large entropic barriers associated with the formation of intra-molecular loops. In this work, we present a simulation scheme that sidesteps configurational costs by dedicated Monte Carlo moves capable of binding/unbinding reactive sites in a single step. Cross-linking reactions are implemented by trial moves that reconstruct chain sections attempting, at the same time, a dimerization reaction between pairs of reactive sites. The model is parametrized by the reaction equilibrium constant of the reactive species free in solution. This quantity can be obtained by means of experiments or atomistic/quantum simulations. We use the proposed methodology to study the self-assembly of single-chain polymeric nanoparticles, starting from flexible precursors carrying regularly or randomly distributed reactive sites. We focus on understanding differences in the morphology of chain nanoparticles when linkages are reversible as compared to the well-studied case of irreversible reactions. Intriguingly, we find that the size of regularly functionalized chains, in good solvent conditions, is non-monotonous as a function of the degree of functionalization. We clarify how this result follows from excluded volume interactions and is peculiar of reversible linkages and regular functionalizations.

  2. Evolving microstructure, magnetic properties and phase transition in a mechanically alloyed Ni0.5Zn0.5Fe2O4 single sample

    Science.gov (United States)

    Ismail, Ismayadi; Hashim, Mansor; Kanagesan, Samikannu; Ibrahim, Idza Riati; Nazlan, Rodziah; Wan Ab Rahman, Wan Norailiana; Abdullah, Nor Hapishah; Mohd Idris, Fadzidah; Bahmanrokh, Ghazaleh; Shafie, Mohd Shamsul Ezzad; Manap, Masni

    2014-02-01

    We report on an investigation to unravel the dependence of magnetic properties on microstructure while they evolve in parallel under the influence of sintering temperature of a single sample of Ni0.5Zn0.5Fe2O4 synthesized via mechanical alloying. A single sample, instead of the normally practiced approach of using multiple samples, was sintered at various sintering temperatures from 500 °C to 1400 °C. The morphology of the samples was studied by means of scanning electron microscopy (SEM) equipped with EDX; density measurement was conducted using the Archimedes principle; and hysteresis measurement was carried out using a B-H hysteresisgraph system. XRD data showed that the first appearance of a single phase was at 800 °C and an amorphous phase was traced at lower sintering temperatures. We correlated the microstructure and the magnetic properties and showed that the important grain-size threshold for the appearance of significant ordered magnetism (mainly ferromagnetism) was about ≥0.3 µm. We found that there were three stages of magnetic phase evolution produced via the sintering process with increasing temperatures. The first stage was dominated by paramagnetic states with some superparamagnetic behavior; the second stage was influenced by moderately ferromagnetic states and some paramagnetic states; and the third stage consisted of strongly ferromagnetic states with negligible paramagnetic states. We found that three factors sensitively influenced the sample's content of ordered magnetism—the ferrite-phase crystallinity degree, the number of grains above the critical grain size and the number of large enough grains for domain wall accommodation.

  3. A single laboratory setup for investigating the anisotropy of both seismic and electrical properties in core samples

    Science.gov (United States)

    David, Christian; Robion, Philippe; Louis, Laurent

    2017-09-01

    A fully automated versatile device is presented for analysing the anisotropy of both seismic and electrical properties on cylindrical rock samples. Initially devoted to the study of P-wave velocity anisotropy (APV), the system has been upgraded to allow the measurement of electrical conductivity anisotropy (AEC) as well. The improved setup allows for the estimation of the APV and AEC, from measurements across multiple diameters with an angular spacing selected by the operator. In order to determine the APV simplified tensor and the true AEC tensor, at least three mutually orthogonal samples are required. Switching from velocity to electrical measurements only requires replacing the pair of ultrasonic transducers by a pair of electrodes. Whereas the procedure is quite straightforward for the P-wave velocity analysis, different calibration steps and approximations are required in the processing of the electrical data. As an example the methodology was applied to a set of two rocks, the Leopard and the Castlegate sandstones, for which the APV and AEC are compared in a unified scheme. Good agreement was found between the orientation of the symmetry axes for both properties, suggesting that in the studied rocks pore space anisotropy accounts for both velocity and electrical anisotropy. This methodology can also be incorporated into a core analysis workflow to obtain a first-hand assessment of the rock fabric orientation and anisotropy with respect to seismic and transport properties. The information thus obtained can be used as a screening step for sampling and stress testing as well as for direct core to log integration and cross-property correlations.

  4. Rapid Treponema pallidum clearance from blood and ulcer samples following single dose benzathine penicillin treatment of early syphilis.

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    Craig Tipple

    2015-02-01

    Full Text Available Currently, the efficacy of syphilis treatment is measured with anti-lipid antibody tests. These can take months to indicate cure and, as a result, syphilis treatment trials require long periods of follow-up. The causative organism, Treponema pallidum (T. pallidum, is detectable in the infectious lesions of early syphilis using DNA amplification. Bacteraemia can likewise be identified, typically in more active disease. We hypothesise that bacterial clearance from blood and ulcers will predict early the standard serology-measured treatment response and have developed a qPCR assay that could monitor this clearance directly in patients with infectious syphilis. Patients with early syphilis were given an intramuscular dose of benzathine penicillin. To investigate the appropriate sampling timeframe samples of blood and ulcer exudate were collected intensively for T. pallidum DNA (tpp047 gene and RNA (16S rRNA quantification. Sampling ended when two consecutive PCRs were negative. Four males were recruited. The mean peak level of T. pallidum DNA was 1626 copies/ml whole blood and the mean clearance half-life was 5.7 hours (std. dev. 0.53. The mean peak of 16S rRNA was 8879 copies/ml whole blood with a clearance half-life of 3.9 hours (std. dev. 0.84. From an ulcer, pre-treatment, 67,400 T. pallidum DNA copies and 7.08 x 107 16S rRNA copies were detected per absorbance strip and the clearance half-lives were 3.2 and 4.1 hours, respectively. Overall, T. pallidum nucleic acids were not detected in any sample collected more than 56 hours (range 20-56 after treatment. All patients achieved serologic cure. In patients with active early syphilis, measuring T. pallidum levels in blood and ulcer exudate may be a useful measure of treatment success in therapeutic trials. These laboratory findings need confirmation on a larger scale and in patients receiving different therapies.

  5. Single-particle characterization of ice-nucleating particles and ice particles residuals sampled by three different techniques

    Science.gov (United States)

    Kandler, Konrad; Worringen, Annette; Benker, Nathalie; Dirsch, Thomas; Mertes, Stephan; Schenk, Ludwig; Kästner, Udo; Frank, Fabian; Nillius, Björn; Bundke, Ulrich; Rose, Diana; Curtius, Joachim; Kupiszewski, Piotr; Weingartner, Ernest; Vochezer, Paul; Schneider, Johannes; Schmidt, Susan; Weinbruch, Stephan; Ebert, Martin

    2015-04-01

    During January/February 2013, at the High Alpine Research Station Jungfraujoch a measurement campaign was carried out, which was centered on atmospheric ice-nucleating particles (INP) and ice particle residuals (IPR). Three different techniques for separation of INP and IPR from the non-ice-active particles are compared. The Ice Selective Inlet (ISI) and the Ice Counterflow Virtual Impactor (Ice-CVI) sample ice particles from mixed phase clouds and allow for the analysis of the residuals. The combination of the Fast Ice Nucleus Chamber (FINCH) and the Ice Nuclei Pumped Counterflow Virtual Impactor (IN-PCVI) provides ice-activating conditions to aerosol particles and extracts the activated INP for analysis. Collected particles were analyzed by scanning electron microscopy and energy-dispersive X-ray microanalysis to determine size, chemical composition and mixing state. All INP/IPR-separating techniques had considerable abundances (median 20 - 70 %) of instrumental contamination artifacts (ISI: Si-O spheres, probably calibration aerosol; Ice-CVI: Al-O particles; FINCH+IN-PCVI: steel particles). Also, potential sampling artifacts (e.g., pure soluble material) occurred with a median abundance of metal oxides were the major INP/IPR particle types separated by all three techniques. Soot was a minor contributor. Lead was detected in less than 10 % of the particles, of which the majority were internal mixtures with other particle types. Sea-salt and sulfates were identified by all three methods as INP/IPR. Most samples showed a maximum of the INP/IPR size distribution at 400 nm geometric diameter. In a few cases, a second super-micron maximum was identified. Soot/carbonaceous material and metal oxides were present mainly in the submicron range. ISI and FINCH yielded silicates and Ca-rich particles mainly with diameters above 1 µm, while the Ice-CVI also separated many submicron IPR. As strictly parallel sampling could not be performed, a part of the discrepancies between

  6. Stool management systems for preventing environmental spread of Clostridium difficile: a comparative trial.

    Science.gov (United States)

    Gray, Mikel; Omar, Amin; Buziak, Brenton

    2014-01-01

    The purpose of this study was to compare contamination of the immediate environment with Clostridium difficile spores and vegetative cells from 2 stool management systems over a period of 30 days in a controlled laboratory setting. In vitro, comparison trial. Two stool management systems were compared over a 30-day period in a controlled laboratory setting. Sixteen systems were filled with sterile loose canine stool inoculated with 10 colony-forming units (CFUs) per milliliter of C difficile; specially prepared culture media were used to detect C difficile contamination on various surfaces of the device and in the immediate environment. Containment bags were changed daily and devices were refilled with inoculated stool to more closely imitate use in the clinical setting. A dichotomous outcome variable (growth vs no growth) was used to analyze contamination on a daily basis via the generalized estimating equation; devices were also compared on days 3, 10, 20, and 30 by measuring CFUs per device surface. Logistic regression analysis was used to analyze growth over time. When observations showed no growth, the Cochran-Mantel Haenszel test was used to compare study devices. Analysis revealed that 20.8% of anterior surfaces of the collection bags for device 1 were contaminated versus 83.9% of collection bags for device 2 (P < .001). Comparison of the tubing/hub interface resulted in similar findings; 20.8% of device 1 group were contaminated versus 86.3% of device 2 group (P < .001). Analysis of an absorbent pad placed under the device during daily changes found that 0.5% of device 1 were contaminated versus 38.1% of pads placed under device 2 (P < .001). Findings from this in vitro study show that stool management systems can limit or prevent environmental contamination of C difficile. Results also reveal significant differences in the 2 systems tested; we hypothesize that these differences are attributable to the interface between the tubing and collection bag, the

  7. Two-dimensional strandness-dependent electrophoresis: a method to characterize single-stranded DNA, double-stranded DNA, and RNA-DNA hybrids in complex samples.

    Science.gov (United States)

    Gunnarsson, Gudmundur H; Gudmundsson, Bjarki; Thormar, Hans G; Alfredsson, Arni; Jonsson, Jon J

    2006-03-01

    We describe two-dimensional strandness-dependent electrophoresis (2D-SDE) for quantification and length distribution analysis of single-stranded (ss) DNA fragments, double-stranded (ds) DNA fragments, RNA-DNA hybrids, and nicked DNA fragments in complex samples. In the first dimension nucleic acid molecules are separated based on strandness and length in the presence of 7 M urea. After the first-dimension electrophoresis all nucleic acid fragments are heat denatured in the gel. During the second-dimension electrophoresis all nucleic acid fragments are single-stranded and migrate according to length. 2D-SDE takes about 90 min and requires only basic skills and equipment. We show that 2D-SDE has many applications in analyzing complex nucleic acid samples including (1) estimation of renaturation efficiency and kinetics, (2) monitoring cDNA synthesis, (3) detection of nicked DNA fragments, and (4) estimation of quality and in vitro damage of nucleic acid samples. Results from 2D-SDE should be useful to validate techniques such as complex polymerase chain reaction, subtractive hybridization, cDNA synthesis, cDNA normalization, and microarray analysis. 2D-SDE could also be used, e.g., to characterize biological nucleic acid samples. Information obtained with 2D-SDE cannot be readily obtained with other methods. 2D-SDE can be used for preparative isolation of ssDNA fragments, dsDNA fragments, and RNA-DNA hybrids.

  8. Simultaneous estimation of population receptive field and hemodynamic parameters from single point BOLD responses using Metropolis-Hastings sampling.

    Science.gov (United States)

    Adaszewski, Stanisław; Slater, David; Melie-Garcia, Lester; Draganski, Bogdan; Bogorodzki, Piotr

    2018-01-30

    We introduce a new approach to Bayesian pRF model estimation using Markov Chain Monte Carlo (MCMC) sampling for simultaneous estimation of pRF and hemodynamic parameters. To obtain high performance on commonly accessible hardware we present a novel heuristic consisting of interpolation between precomputed responses for predetermined stimuli and a large cross-section of receptive field parameters. We investigate the validity of the proposed approach with respect to MCMC convergence, tuning and biases. We compare different combinations of pRF - Compressive Spatial Summation (CSS), Dumoulin-Wandell (DW) and hemodynamic (5-parameter and 3-parameter Balloon-Windkessel) models within our framework with and without the usage of the new heuristic. We evaluate estimation consistency and log probability across models. We perform as well a comparison of one model with and without lookup table within the RStan framework using its No-U-Turn Sampler. We present accelerated computation of whole-ROI parameters for one subject. Finally, we discuss risks and limitations associated with the usage of the new heuristic as well as the means of resolving them. We found that the new algorithm is a valid sampling approach to joint pRF/hemodynamic parameter estimation and that it exhibits very high performance. Copyright © 2018. Published by Elsevier Inc.

  9. Single-particle characterization of ice-nucleating particles and ice particle residuals sampled by three different techniques

    Science.gov (United States)

    Worringen, A.; Kandler, K.; Benker, N.; Dirsch, T.; Mertes, S.; Schenk, L.; Kästner, U.; Frank, F.; Nillius, B.; Bundke, U.; Rose, D.; Curtius, J.; Kupiszewski, P.; Weingartner, E.; Vochezer, P.; Schneider, J.; Schmidt, S.; Weinbruch, S.; Ebert, M.

    2015-04-01

    In the present work, three different techniques to separate ice-nucleating particles (INPs) as well as ice particle residuals (IPRs) from non-ice-active particles are compared. The Ice Selective Inlet (ISI) and the Ice Counterflow Virtual Impactor (Ice-CVI) sample ice particles from mixed-phase clouds and allow after evaporation in the instrument for the analysis of the residuals. The Fast Ice Nucleus Chamber (FINCH) coupled with the Ice Nuclei Pumped Counterflow Virtual Impactor (IN-PCVI) provides ice-activating conditions to aerosol particles and extracts the activated particles for analysis. The instruments were run during a joint field campaign which took place in January and February 2013 at the High Alpine Research Station Jungfraujoch (Switzerland). INPs and IPRs were analyzed offline by scanning electron microscopy and energy-dispersive X-ray microanalysis to determine their size, chemical composition and mixing state. Online analysis of the size and chemical composition of INP activated in FINCH was performed by laser ablation mass spectrometry. With all three INP/IPR separation techniques high abundances (median 20-70%) of instrumental contamination artifacts were observed (ISI: Si-O spheres, probably calibration aerosol; Ice-CVI: Al-O particles; FINCH + IN-PCVI: steel particles). After removal of the instrumental contamination particles, silicates, Ca-rich particles, carbonaceous material and metal oxides were the major INP/IPR particle types obtained by all three techniques. In addition, considerable amounts (median abundance mostly a few percent) of soluble material (e.g., sea salt, sulfates) were observed. As these soluble particles are often not expected to act as INP/IPR, we consider them as potential measurement artifacts. Minor types of INP/IPR include soot and Pb-bearing particles. The Pb-bearing particles are mainly present as an internal mixture with other particle types. Most samples showed a maximum of the INP/IPR size distribution at 200

  10. Evaluation of a QIAamp DNA stool purification kit for Shiga-toxigenic Escherichia coli detection in bovine fecal swabs by PCR Evaluación del kit QIAamp DNA stool purification para la detección de Escherichia coli productor de toxina Shiga en hisopados de materia fecal bovina por PCR[

    Directory of Open Access Journals (Sweden)

    A. Gioffré

    2004-03-01

    Full Text Available A commercial kit intended for Taq polymerase inhibitor removal was tested to detect Shiga-toxigenic Escherichia coli (STEC by polymersase chain reaction (PCR directly from cattle fecal samples. Forty-five samples were analysed for the presence of stx genes. Results were compared to those obtained by two other methods: amplification of DNA purified by a non-commercial procedure (heat lysis protocol, and amplification of DNA from samples cultured in solid media, commonly used in our lab. Identical numbers of positive samples (33/45, 73 % were obtained with the QIAamp DNA stool purification kit and the culturing procedure, suggesting an adequate removal of inhibitors that interfere in PCR amplification from the feces. Besides, the number of positive samples detected using DNA purified by the non-commercial protocol was lower, 25/39 (64% than that achieved by using the kit. In conclusion, the use of the QIAamp DNA stool purification kit provided a rapid stx gene detection by PCR in bovine fecal samples.Un kit comercial diseñado para la eliminación de inhibidores de la polimerasa Taq fue ensayado para la detección de STEC por PCR en muestras fecales de bovinos. Cuarenta y cinco muestras fueron evaluadas por la presencia de genes stx. Los resultados fueron comparados con aquéllos obtenidos por otros dos métodos: amplificación de ADN purificado por un procedimiento no comercial (protocolo de lisis por calor, y amplificación de ADN de muestras cultivadas en medio sólido, comúnmente usado en nuestro laboratorio. El mismo número de muestras positivas (33/45, 73 %, fueron obtenidas con el QIAamp DNA stool purification kit y el procedimiento de cultivo, sugiriendo una eliminación adecuada de inhibidores que interfieren con la amplificación en materia fecal. Por otro lado, el número de muestras positivas detectadas usando ADN purificado por el protocolo no comercial fue menor, 25/39 (64%. En conclusión, el uso del kit QIAamp DNA stool

  11. Ruthenium(II) piano stool coordination compounds with aminomethylphosphanes: Synthesis, characterisation and preliminary biological study in vitro.

    Science.gov (United States)

    Płotek, Michał; Starosta, Radosław; Komarnicka, Urszula K; Skórska-Stania, Agnieszka; Kołoczek, Przemysław; Kyzioł, Agnieszka

    2017-05-01

    Reaction of {[Ru(η 6 -p-cymene)Cl] 2 (μ-Cl) 2 } (1) with aminomethylphosphane derived from morpholine (P{CH 2 N(CH 2 CH 2 ) 2 O} 3 (A), PPh 2 {CH 2 N(CH 2 CH 2 ) 2 O} (B)) or piperazine (P{CH 2 N(CH 2 CH 2 ) 2 NCH 2 CH 3 } 3 (C), PPh 2 {CH 2 N(CH 2 CH 2 ) 2 NCH 2 CH 3 } (D)) results in four new piano stool ruthenium(II) coordination compounds: [Ru(η 6 -p-cymene)Cl 2 (A)] (2A), [Ru(η 6 -p-cymene)Cl 2 (B)] (2B), [Ru(η 6 -p-cymene)Cl 2 (C)] (2C) and [Ru(η 6 -p-cymene)Cl 2 (D)] (2D). Every complex was fully characterized using spectroscopic methods ( 1 H, 13 C{ 1 H}, 31 P{ 1 H} NMR and ESI-MS), elemental analysis, X-ray single crystal diffraction and DFT calculations. Preliminary studies of in vitro cytotoxicity on the A549 (human lung adenocarcinoma) and MCF7 (human breast adenocarcinoma) cell lines revealed 2A-2D activity in the same order of magnitude as in the case of cisplatin. Additionally, the study confirmed the ability of 2A-2D to interact with DNA helix and transferrin. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Application of a stool antigen test to evaluate the burden of Helicobacter pylori infection in dyspepsia patients

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    Rumpa Saha

    2016-01-01

    Full Text Available Helicobacter pylori (HP is causally associated with peptic ulcer disease and gastric carcinoma. Determination of the prevalence of HP infection in dyspepsia patients′ in particular geographical area is imperative for the appropriate management of dyspepsia. HP antigen detection in stool is a noninvasive diagnostic test of HP infection. This prospective study was conducted to find out the prevalence of HP infection based on stool antigen testing in dyspeptic patients who had also undergone upper gastrointestinal (GI endoscopy. This study highlights the high prevalence of HP infection in dyspeptic Indian patients, particularly males, and emphasizes the growing importance of the bacterium causing infection among children. We also found HP stool antigen testing to be superior to upper GI endoscopy for detecting HP infection. Hence, we recommend initial testing for HP stool antigen in dyspeptic patients before initiating treatment and before carrying out any invasive procedure such as endoscopy.

  13. A Single-Chip 64-Channel Ultrasound RX-Beamformer Including Analog Front-End and an LUT for Non-Uniform ADC-Sample-Clock Generation.

    Science.gov (United States)

    Kim, Yoon-Jee; Cho, Sung-Eun; Um, Ji-Yong; Chae, Min-Kyun; Bang, Jihoon; Song, Jongkeun; Jeon, Taeho; Kim, Byungsub; Sim, Jae-Yoon; Park, Hong-June

    2017-02-01

    A 64-channel RX digital beamformer was implemented in a single chip for 3-D ultrasound medical imaging using 2-D phased-array transducers. The RX beamformer chip includes 64 analog front-end branches including 64 non-uniform sampling ADCs, a FIFO/Adder, and an on-chip look-up table (LUT). The LUT stores the information on the rising edge timing of the non-uniform ADC sampling clocks. To include the LUT inside the beamformer chip, the LUT size was reduced by around 240 times by approximating an ADC-sample-time profile w.r.t. focal points (FP) along a scanline (SL) for a channel into a piece-wise linear form. The maximum error between the approximated and accurate sample times of ADC is eight times the sample time resolution (Ts) that is 1/32 of the ultrasound signal period in this work. The non-uniform sampling reduces the FIFO size required for digital beamforming by around 20 times. By applying a 9-dot image from Field-II program and 2-D ultrasound phantom images to the fabricated RX beamformer chip, the original images were successfully reconstructed from the measured output. The chip in a 0.13-um CMOS occupies 30.25 [Formula: see text] and consumes 605 mW.

  14. Utility of a stool antigen test to detect the incidence of helicobacter pylori infection and familial and community enviromental risk factors for this infection in pediatric age

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    T. Sabbi

    2012-04-01

    Full Text Available Background: helicobacter pylori (hp infection is mainly acquired during childhood; it is recognised as a cause of gastritis and peptic ulcer and it has been classified as a group A carcinogen by World health organization. the exact mode of transmission is as yet, not known. Aim of our study has been to identify risk factors associated with helicobacter pylori infection in a preschool and school population and to confirm if hp antigen in faeces is useful as screening in epidemiological studies. Methods: We interviewed, with questionnaire, 400 children (203 male; age range 3-10 years; mean age 6 years of 3 different schools and stool samples were collected of all children too. 35 of 400 (8% children underwent to upper gastrointestinal endoscopy because of a suspect of upper gastrointestinal disease. Results: stool were collected from 400 school children and 35 of them shown positivity of hp antigen test. A questionnaire about presence of nausea, vomit, recurrent abdominal pain, family size, parent’s occupations and education, use of antibiotics, country of birth of child and parents, personal hygiene, breast feeding, presence of the animals was completed. 35 children with positive hp stool antigen test and a suspicious of upper gastrointestinal disease (recurrent abdominal pain, diurnal or nocturnal abdominal pain, nausea, vomiting, iron deficiency underwent to esophagogastroduodenoscopy (egdS that demonstrated antral gastritis and positive histology and urease rapid test. Conclusions: the results of this study suggest that risk factors for hp infection are low socioeconomics factors, hygiene and living conditions and that hp antigen in faeces is useful as screening test.

  15. Prevalence and antibiotic susceptibility pattern of floroquinolone resistant S h i g e l l a species isolated from infants stool in Gulbarga district, Karnataka, India

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    Prabhurajeshwar

    2015-06-01

    Full Text Available Objective: To resolve the incidence rate of Shigella species and emergence of multiple drug resistance to cephalosporins and fluoroquinolones generation drugs in Gulbarga district and nearby area is a matter of concern for the better management of Shigellosis in children. Methods: The study work involved all infants with acute diarrhoea, ranging 1 month to 5 years of age. Total 334 stool samples were collected during 2013-2014. Incidence, phenotypic and genotypic (16S rRNA characteristics and antimicrobial resistance were determined for emerging antibiotics against Shigella strains were studied. Results: Total 43 (12.87% positive Shigella species retrieved from 334 stool samples, Shigella dysentriae (48.83%, Shigella flexneri (32.55% were the predominant, followed by Shigella sonnei (18.60% and no case of Shigella boydii (0%. Among the Shigella isolates 32 (74.41% were MDR strains, confer the high resistance rate like 100%, 95.33%, 87.5% and 85% to different class of antibiotics studied. Conclusions: This study illustrates the appearance of the results of long-term supervision and antimicrobial resistance in clinical Shigella strains. Data collected as part of the examination will be involved for developing treatment, allocate for the community and to present control report with which to discriminate outbreak strains in the future.

  16. Recombinase Polymerase Amplification Compared to Real-Time Polymerase Chain Reaction Test for the Detection of Fasciola hepatica in Human Stool

    Science.gov (United States)

    Cabada, Miguel M.; Malaga, Jose L.; Castellanos-Gonzalez, Alejandro; Bagwell, Kelli A.; Naeger, Patrick A.; Rogers, Hayley K.; Maharsi, Safa; Mbaka, Maryann; White, A. Clinton

    2017-01-01

    Fasciola hepatica is the most widely distributed trematode infection in the world. Control efforts may be hindered by the lack of diagnostic capacity especially in remote endemic areas. Polymerase chain reaction (PCR)–based methods offer high sensitivity and specificity but require expensive technology. However, the recombinase polymerase amplification (RPA) is an efficient isothermal method that eliminates the need for a thermal cycler and has a high deployment potential to resource-limited settings. We report on the characterization of RPA and PCR tests to detect Fasciola infection in clinical stool samples with low egg burdens. The sensitivity of the RPA and PCR were 87% and 66%, respectively. Both tests were 100% specific showing no cross-reactivity with trematode, cestode, or nematode parasites. In addition, RPA and PCR were able to detect 47% and 26% of infections not detected by microscopy, respectively. The RPA adapted to a lateral flow platform was more sensitive than gel-based detection of the reaction products. In conclusion, the Fasciola RPA is a highly sensitive and specific test to diagnose chronic infection using stool samples. The Fasciola RPA lateral flow has the potential for deployment to endemic areas after further characterization. PMID:27821691

  17. A one-step immune-chromatographic Helicobacter pylori stool antigen test for children was quick, consistent, reliable and specific.

    Science.gov (United States)

    Kalach, Nicolas; Gosset, Pierre; Dehecq, Eric; Decoster, Anne; Georgel, Anne-France; Spyckerelle, Claire; Papadopoulos, Stephanos; Dupont, Christophe; Raymond, Josette

    2017-12-01

    This French study assessed a quick, noninvasive, immuno-chromatographic, Helicobacter pylori (H. pylori) stool antigen test for detecting infections in children. We enrolled 158 children, with a median age of 8.5 years (range eight months to 17 years), with digestive symptoms suggesting upper gastrointestinal tract disease. Upper digestive endoscopy was performed with gastric biopsy specimens for histology, a rapid urease test, culture test and quantitative real-time polymerase chain reaction. The H. pylori stool antigen test was performed twice for each child and the results were compared to the reference method. The reference methods showed that 23 (14.6%) of the 158 children tested were H. pylori positive. The H. pylori stool antigen test showed 91.3% sensitivity, with a 95% confidence interval (95% CI) of 86.9-95.6 and 97% specificity (95% CI 94.3-99.6), 30.84 positive likelihood ratio and 0.09 negative likelihood ratio. The test accuracy was 96.2% (95% CI 93.2-99.1). The two blinded independent observers produced identical H. pylori stool antigen test results and the Kappa coefficient for the H. pylori stool antigen test was one. The H. pylori stool antigen test was found to be a consistent, reliable, quick and specific test for detecting the H. pylori infection in children. ©2017 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  18. Magnetic resonance colonography without bowel cleansing using oral and rectal stool softeners (fecal cracking) - a feasibility study

    International Nuclear Information System (INIS)

    Ajaj, Waleed; Lauenstein, Thomas C.; Kuehle, Christiane; Herborn, Christoph U.; Goehde, Susanne C.; Schneemann, Hubert; Ruehm, Stefan G.; Goyen, Mathias

    2005-01-01

    The aim of our study was to assess the effect of oral and rectal stool softeners on dark-lumen magnetic resonance (MR) colonography without bowel cleansing. Ten volunteers underwent MR colonography without colonic cleansing. A baseline examination was performed without oral or rectal administration of stool softeners. In a second set, volunteers ingested 60 ml of lactulose 24 h prior to MR examination. In a third examination, water as a rectal enema was replaced by a solution of 0.5%-docusate sodium (DS). A fourth MR examination was performed, in conjunction with both oral administration of lactulose and rectal application of DS. A T1-weighted data set was acquired at scanning times of 0, 5 and 10 min after colonic filling. A fourth data set was acquired 75 s after i.v. injection of contrast agent. Signal intensity of stool was calculated for all colonic segments. Without oral ingestion of lactulose or rectal enema with DS stool signal intensity was high and did not decrease over time. However, lactulose and DS caused a decrease in stool signal intensity. Both substances together led to a decreasing signal intensity of feces. Combination of lactulose and DS provided the lowest signal intensity of stool. Thus, feces could hardly be distinguished from dark rectal enema allowing for the assessment of the colonic wall. (orig.)

  19. Non-Destructive Study of Bulk Crystallinity and Elemental Composition of Natural Gold Single Crystal Samples by Energy-Resolved Neutron Imaging

    Science.gov (United States)

    Tremsin, Anton S.; Rakovan, John; Shinohara, Takenao; Kockelmann, Winfried; Losko, Adrian S.; Vogel, Sven C.

    2017-01-01

    Energy-resolved neutron imaging enables non-destructive analyses of bulk structure and elemental composition, which can be resolved with high spatial resolution at bright pulsed spallation neutron sources due to recent developments and improvements of neutron counting detectors. This technique, suitable for many applications, is demonstrated here with a specific study of ~5–10 mm thick natural gold samples. Through the analysis of neutron absorption resonances the spatial distribution of palladium (with average elemental concentration of ~0.4 atom% and ~5 atom%) is mapped within the gold samples. At the same time, the analysis of coherent neutron scattering in the thermal and cold energy regimes reveals which samples have a single-crystalline bulk structure through the entire sample volume. A spatially resolved analysis is possible because neutron transmission spectra are measured simultaneously on each detector pixel in the epithermal, thermal and cold energy ranges. With a pixel size of 55 μm and a detector-area of 512 by 512 pixels, a total of 262,144 neutron transmission spectra are measured concurrently. The results of our experiments indicate that high resolution energy-resolved neutron imaging is a very attractive analytical technique in cases where other conventional non-destructive methods are ineffective due to sample opacity. PMID:28102285

  20. Isolation and cloning of exendin precursor cDNAs from single samples of venom from the Mexican beaded lizard (Heloderma horridum) and the Gila monster (Heloderma suspectum).

    Science.gov (United States)

    Chen, Tianbao; Kwok, Hangfai; Ivanyi, Craig; Shaw, Chris

    2006-03-01

    Reptile venoms are complex cocktails of bioactive molecules, including peptides. While the drug discovery potential of most species remains unrealized, many are endangered and afforded protection under international treaties. In this study, we describe how potential clinically important bioactive peptides and their corresponding mRNAs can be structurally characterized from single, small samples of reptile venom. The potential type-2 diabetes therapeutics, exendin-3 and exendin-4, from the Mexican beaded lizard (Heloderma horridum) and the Gila monster (Heloderma suspectum), respectively, have been characterized at both protein and nucleic acid levels to illustrate the efficacy of the technique and its contribution to biodiversity conservation.

  1. Relative effectiveness of kinetic analysis vs single point readings for classifying environmental samples based on community-level physiological profiles (CLPP)

    Science.gov (United States)

    Garland, J. L.; Mills, A. L.; Young, J. S.

    2001-01-01

    The relative effectiveness of average-well-color-development-normalized single-point absorbance readings (AWCD) vs the kinetic parameters mu(m), lambda, A, and integral (AREA) of the modified Gompertz equation fit to the color development curve resulting from reduction of a redox sensitive dye from microbial respiration of 95 separate sole carbon sources in microplate wells was compared for a dilution series of rhizosphere samples from hydroponically grown wheat and potato ranging in inoculum densities of 1 x 10(4)-4 x 10(6) cells ml-1. Patterns generated with each parameter were analyzed using principal component analysis (PCA) and discriminant function analysis (DFA) to test relative resolving power. Samples of equivalent cell density (undiluted samples) were correctly classified by rhizosphere type for all parameters based on DFA analysis of the first five PC scores. Analysis of undiluted and 1:4 diluted samples resulted in misclassification of at least two of the wheat samples for all parameters except the AWCD normalized (0.50 abs. units) data, and analysis of undiluted, 1:4, and 1:16 diluted samples resulted in misclassification for all parameter types. Ordination of samples along the first principal component (PC) was correlated to inoculum density in analyses performed on all of the kinetic parameters, but no such influence was seen for AWCD-derived results. The carbon sources responsible for classification differed among the variable types with the exception of AREA and A, which were strongly correlated. These results indicate that the use of kinetic parameters for pattern analysis in CLPP may provide some additional information, but only if the influence of inoculum density is carefully considered. c2001 Elsevier Science Ltd. All rights reserved.

  2. The effect of changing stool collection processes on compliance in nationwide organized screening using a fecal occult blood test (FOBT) in Korea: study protocol for a randomized controlled trial.

    Science.gov (United States)

    Shin, Hye Young; Suh, Mina; Baik, Hyung Won; Choi, Kui Son; Park, Boyoung; Jun, Jae Kwan; Lee, Chan Wha; Oh, Jae Hwan; Lee, You Kyoung; Han, Dong Soo; Lee, Do-Hoon

    2014-11-26

    Colorectal cancer (CRC) screening by fecal occult blood test (FOBT) significantly reduces CRC mortality, and compliance rates directly influence the efficacy of this screening method. The aim of this study is to investigate whether stool collection strategies affect compliance with the FOBT. In total, 3,596 study participants aged between 50 and 74 years will be recruited. The study will be conducted using a randomized controlled trial, with a 2 × 2 factorial design resulting in four groups. The first factor is the method of stool-collection device distribution (mailing vs. visiting the clinic) and the second is the type of stool-collection device (sampling kit vs. conventional container). Participants will be randomly assigned to one of four groups: (1) sampling kit received by mail; (2) conventional container received by mail; (3) sampling kit received at the clinic; (4) conventional container received at the clinic (control group). The primary outcome will be the FOBT compliance rate; satisfaction and intention to be rescreened in the next screening round will also be evaluated. The rates of positive FOBT results and detection of advanced adenomas or cancers through colonoscopies will also be compared between the two collection containers. Identifying a method of FOBT that yields high compliance rates will be a key determinant of the success of CRC screening. The findings of this study will provide reliable information for health policy makers to develop evidence-based strategies for a high compliance rate. KCT0000803 Date of registration in primary registry: 9 January, 2013.

  3. Results of a screening method used in a 12-month stool survey for Escherichia coli O157:H7.

    Science.gov (United States)

    Harris, A A; Kaplan, R L; Goodman, L J; Doyle, M; Landau, W; Segreti, J; Mayer, K; Levin, S

    1985-10-01

    Escherichia coli serotype O157:H7 has been epidemiologically linked to outbreaks of hemorrhagic colitis associated with fast-food restaurants and nursing homes. Sporadic cases now exceed those associated with outbreaks. The incidence of the organism in patients with common diarrhea syndromes and in asymptomatic persons is unknown. Routine serotyping of E. coli isolates is impractical for most clinical microbiology laboratories. We developed a screening plate by utilizing sorbitol fermentation as a biochemical marker to identify organisms for serotyping. A total of 2,552 stool samples were screened. In 106 (4.1%), sorbitol-negative E. coli were identified. Of these, two were serotype O157:H7, and both produced a Vero cell toxin. One patient had hemorrhagic colitis and the other a mild, febrile, self-limited diarrhea with no other bacterial pathogen identified. This plate provides an easy, effective method of screening for sorbitol-negative E. coli, a process facilitating the selection of organisms for serotyping and one that may help clarify this organism's role in human disease.

  4. A precise evaluation of glomerular filtration rate (GFR) in two plasma samples following a single administration of 57Co-B12 vitamin

    International Nuclear Information System (INIS)

    Camargo, E.E.; Rockmann, R.L.; Barreto, T.M.; Eston, T.E.; Papaleo Netto, M.; Carvalho, N.

    1974-01-01

    Through a logarithmic regression performed with the contings of 4 plasma samples withdrawn at 20,40,60 and 80 minutes after a venous injection of vitamin B 12 - 57 Co, the glomerular filtration-rate(GFR) in 11 patients, performing simultaneously the same study with EDTA- 51 Cr in 3 of them, is evaluated. The values obtained through the regression straight line are compared with those given by only 2 points, in the 6 possible combinations: 20 and 40 minutes, 20 and 60 minutes, 20 and 80 minutes, 40 and 60 minutes, 40 and 80 minutes, 60 and 80 minutes. The pair of points obtained at 20 and 80 minutes determined the straight line most similar to the logarithmic regression and as a simplification of the method, the withdraw of only 2 plasma samples, at and 80 minutes after a single injection of vitamin B 12 -57 Co is proposed [pt

  5. Association Between Stool Enteropathogen Quantity and Disease in Tanzanian Children Using TaqMan Array Cards: A Nested Case-Control Study

    Science.gov (United States)

    Platts-Mills, James A.; Gratz, Jean; Mduma, Esto; Svensen, Erling; Amour, Caroline; Liu, Jie; Maro, Athanasia; Saidi, Queen; Swai, Ndealilia; Kumburu, Happiness; McCormick, Benjamin J. J.; Kibiki, Gibson; Houpt, Eric R.

    2014-01-01

    Etiologic studies of diarrhea are limited by uneven diagnostic methods and frequent asymptomatic detection of enteropathogens. Polymerase chain reaction-based stool pathogen quantification may help distinguish clinically significant infections. We performed a nested case-control study of diarrhea in infants from a community-based birth cohort in Tanzania. We tested 71 diarrheal samples and pre-diarrheal matched controls with a laboratory-developed TaqMan Array Card for 19 enteropathogens. With qualitative detection, no pathogens were significantly associated with diarrhea. When pathogen quantity was considered, rotavirus (odds ratio [OR] = 2.70 per log10 increase, P < 0.001), astrovirus (OR = 1.49, P = 0.01), and Shigella/enteroinvasive Escherichia coli (OR = 1.47, P = 0.04) were associated with diarrhea. Enterotoxigenic E. coli (0.15 SD decline in length-for-age z score after 3 months per log10 increase, P < 0.001) and Campylobacter jejuni/C. coli (0.11 SD decline, P = 0.003) in pre-diarrheal stools were associated with poor linear growth. Quantitative analysis can help refine the association between enteropathogens and disease in endemic settings. PMID:24189366

  6. Reversed-phase single drop microextraction followed by high-performance liquid chromatography with fluorescence detection for the quantification of synthetic phenolic antioxidants in edible oil samples.

    Science.gov (United States)

    Farajmand, Bahman; Esteki, Mahnaz; Koohpour, Elham; Salmani, Vahid

    2017-04-01

    The reversed-phase mode of single drop microextraction has been used as a preparation method for the extraction of some phenolic antioxidants from edible oil samples. Butylated hydroxyl anisole, tert-butylhydroquinone and butylated hydroxytoluene were employed as target compounds for this study. High-performance liquid chromatography followed by fluorescence detection was applied for final determination of target compounds. The most interesting feature of this study is the application of a disposable insulin syringe with some modification for microextraction procedure that efficiently improved the volume and stability of the solvent microdrop. Different parameters such as the type and volume of solvent, sample stirring rate, extraction temperature, and time were investigated and optimized. Analytical performances of the method were evaluated under optimized conditions. Under the optimal conditions, relative standard deviations were between 4.4 and 10.2%. Linear dynamic ranges were 20-10 000 to 2-1000 μg/g (depending on the analytes). Detection limits were 5-670 ng/g. Finally, the proposed method was successfully used for quantification of the antioxidants in some edible oil samples prepared from market. Relative recoveries were achieved from 88 to 111%. The proposed method had a simplicity of operation, low cost, and successful application for real samples. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Relationship between single and multiple perpetrator rape perpetration in South Africa: A comparison of risk factors in a population-based sample.

    Science.gov (United States)

    R, Jewkes; Y, Sikweyiya; K, Dunkle; R, Morrell

    2015-07-07

    Studies of rape of women seldom distinguish between men's participation in acts of single and multiple perpetrator rape. Multiple perpetrator rape (MPR) occurs globally with serious consequences for women. In South Africa it is a cultural practice with defined circumstances in which it commonly occurs. Prevention requires an understanding of whether it is a context specific intensification of single perpetrator rape, or a distinctly different practice of different men. This paper aims to address this question. We conducted a cross-sectional household study with a multi-stage, randomly selected sample of 1686 men aged 18-49 who completed a questionnaire administered using an Audio-enhanced Personal Digital Assistant. We attempted to fit an ordered logistic regression model for factors associated with rape perpetration. 27.6 % of men had raped and 8.8 % had perpetrated multiple perpetrator rape (MPR). Thus 31.9 % of men who had ever raped had done so with other perpetrators. An ordered regression model was fitted, showing that the same associated factors, albeit at higher prevalence, are associated with SPR and MPR. Multiple perpetrator rape appears as an intensified form of single perpetrator rape, rather than a different form of rape. Prevention approaches need to be mainstreamed among young men.

  8. Current sharing temperature of NbTi SULTAN samples compared to prediction using a single pinning mechanism parametrization for NbTi strand

    International Nuclear Information System (INIS)

    Pong, Ian; Vostner, Alexander; Devred, Arnaud; Bessette, Denis; Mitchell, Neil; Bordini, Bernardo; Bottura, Luca; Jewell, Matthew; Long Feng; Wu Yu

    2012-01-01

    NbTi strands to be used in four of the six ITER poloidal field (PF) coils, all the correction coils (CC) and all the superconducting feeder busbars are being produced in China. Short full-size qualification conductor (cabled and jacketed) samples have been developed at ASIPP and tested at CRPP. Single pinning mechanism parametrization for this Chinese strand (type S2) has been obtained using the Bottura scaling law. The determination of the scaling parameters using a Kramer-type regression method will be described. A comparison between the critical temperature at the operating current and field of a single strand as determined by the parametrization and the current sharing temperature (T CS ) of a few conductor samples tested at the SULTAN facility will be made. The validity and limitation of the estimation will be discussed. The estimated T CS dependence on various (superconducting critical as well as geometric and volumetric) parameters will be assessed using the modelled critical surface. Errors propagated from critical current (I c ) measurements of the strands and parameter fitting, and other uncertainties, will be quantified. (paper)

  9. Replica exchange enveloping distribution sampling (RE-EDS): A robust method to estimate multiple free-energy differences from a single simulation.

    Science.gov (United States)

    Sidler, Dominik; Schwaninger, Arthur; Riniker, Sereina

    2016-10-21

    In molecular dynamics (MD) simulations, free-energy differences are often calculated using free energy perturbation or thermodynamic integration (TI) methods. However, both techniques are only suited to calculate free-energy differences between two end states. Enveloping distribution sampling (EDS) presents an attractive alternative that allows to calculate multiple free-energy differences in a single simulation. In EDS, a reference state is simulated which "envelopes" the end states. The challenge of this methodology is the determination of optimal reference-state parameters to ensure equal sampling of all end states. Currently, the automatic determination of the reference-state parameters for multiple end states is an unsolved issue that limits the application of the methodology. To resolve this, we have generalised the replica-exchange EDS (RE-EDS) approach, introduced by Lee et al. [J. Chem. Theory Comput. 10, 2738 (2014)] for constant-pH MD simulations. By exchanging configurations between replicas with different reference-state parameters, the complexity of the parameter-choice problem can be substantially reduced. A new robust scheme to estimate the reference-state parameters from a short initial RE-EDS simulation with default parameters was developed, which allowed the calculation of 36 free-energy differences between nine small-molecule inhibitors of phenylethanolamine N-methyltransferase from a single simulation. The resulting free-energy differences were in excellent agreement with values obtained previously by TI and two-state EDS simulations.

  10. Randomized controlled trial: Standard versus supplemental bowel preparation in patients with Bristol stool form 1 and 2.

    Directory of Open Access Journals (Sweden)

    Yueyue Li

    Full Text Available Bristol stool form 1 and 2 is an important predictor of inadequate bowel preparation.To evaluate the efficacy of supplemental preparation in bowel cleansing quality among patients with Bristol stool form 1 and 2, as well as the feasibility of tailored bowel preparation guided by Bristol stool form scale.Patients with Bristol stool form 1 and 2 from 3 Chinese tertiary hospitals randomly received either 2 L PEG-ELP (group A or 10 mg bisacodyl plus 2 L PEG-ELP (group B; patients with Bristol stool form 3 to 7 received 2 L PEG-ELP (group C for bowel preparation. The primary endpoint is the rate of adequate bowel reparation for the whole colon. The adequate bowel preparation rate for separate colon segments, the polyp detection rate (PDR, tolerability, acceptability, sleeping quality and compliance were evaluated as secondary endpoints.700 patients were randomized. In per-protocol analysis, patients in group B attained significantly higher successful preparation rate than group A (88.7% vs. 61.2%, p<0.001 and similar with group C (88.7% vs. 85.0%, p = 0.316. The PDR in group B was significantly higher than group A (43.2% vs. 25.7%, p<0.001. Acceptability was much higher in group B and C.10 mg bisacodyl plus 2 L PEG-ELP can significantly improve both bowel preparation quality and PDR in patients with Bristol stool form 1 and 2. Bristol stool form scale may be an easy and efficient guide for tailored bowel preparation before colonoscopy.

  11. Simultaneous determination of PPCPs, EDCs, and artificial sweeteners in environmental water samples using a single-step SPE coupled with HPLC-MS/MS and isotope dilution.

    Science.gov (United States)

    Tran, Ngoc Han; Hu, Jiangyong; Ong, Say Leong

    2013-09-15

    A high-throughput method for the simultaneous determination of 24 pharmaceuticals and personal care products (PPCPs), endocrine disrupting chemicals (EDCs) and artificial sweeteners (ASs) was developed. The method was based on a single-step solid phase extraction (SPE) coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and isotope dilution. In this study, a single-step SPE procedure was optimized for simultaneous extraction of all target analytes. Good recoveries (≥ 70%) were observed for all target analytes when extraction was performed using Chromabond(®) HR-X (500 mg, 6 mL) cartridges under acidic condition (pH 2). HPLC-MS/MS parameters were optimized for the simultaneous analysis of 24 PPCPs, EDCs and ASs in a single injection. Quantification was performed by using 13 isotopically labeled internal standards (ILIS), which allows correcting efficiently the loss of the analytes during SPE procedure, matrix effects during HPLC-MS/MS and fluctuation in MS/MS signal intensity due to instrument. Method quantification limit (MQL) for most of the target analytes was below 10 ng/L in all water samples. The method was successfully applied for the simultaneous determination of PPCPs, EDCs and ASs in raw wastewater, surface water and groundwater samples collected in a local catchment area in Singapore. In conclusion, the developed method provided a valuable tool for investigating the occurrence, behavior, transport, and the fate of PPCPs, EDCs and ASs in the aquatic environment. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. [Determination of bile acids in stools of patients with colonic neoplasms and adenomatous polyps].

    Science.gov (United States)

    Paniagua Estévez, M; Roque Lozano, J; Cerdán Cordoví, A; Rodríguez Miranda, A

    1994-01-01

    Since several years ago, the biliar acids have been incriminated in the etiopathogeny of colon cancer and adenomatous polyps, above all the secondary ones, involved by its aggressive action over the colonic epithelium in these mechanisms. The dietetical habits of developed countries have the high responsibility for this situation, their food pattern being a high animal fat diet, high in refined carbohydrates, animal proteins and low in dietetic fiber (diet type "occidental") unlike to developing countries that have a high natural fiber diet, having a much lower incidence in colon cancer and adenomatous polyps. Dietetic fiber has been studied considering it with a protector effect over the aggressive action of biliar acids on the colon mucous. We have studied 60 patients, 20 of them with colon cancer, 20 with adenomatous and 20 case controls without colonic pathology. All of them had total high biliar acids in stools, a dietetical screening was carried out to determine the intake of animal fat and dietetic fibre during a week. There was a significant correlation in cases of cancer, polyps and biliar acids high in stools. There was also a significant correlation between the undue dietetic habits in colon cancer patients and high bilar acids. In those cases of adenomatous polyps, there was not a significant relation to dietetic habits.

  13. Oral Supplementation with Bovine Colostrum Decreases Intestinal Permeability and Stool Concentrations of Zonulin in Athletes

    Directory of Open Access Journals (Sweden)

    Maciej Hałasa

    2017-04-01

    Full Text Available Increased intestinal permeability has been implicated in various pathologies, has various causes, and can develop during vigorous athletic training. Colostrum bovinum is a natural supplement with a wide range of supposed positive health effects, including reduction of intestine permeability. We assessed influence of colostrum supplementation on intestinal permeability related parameters in a group of 16 athletes during peak training for competition. This double-blind placebo-controlled study compared supplementation for 20 days with 500 mg of colostrum bovinum or placebo (whey. Gut permeability status was assayed by differential absorption of lactulose and mannitol (L/M test and stool zonulin concentration. Baseline L/M tests found that six of the participants (75% in the colostrum group had increased intestinal permeability. After supplementation, the test values were within the normal range and were significantly lower than at baseline. The colostrum group Δ values produced by comparing the post-intervention and baseline results were also significantly lower than the placebo group Δ values. The differences in stool zonulin concentration were smaller than those in the L/M test, but were significant when the Δ values due to intervention were compared between the colostrum group and the placebo group. Colostrum bovinum supplementation was safe and effective in decreasing of intestinal permeability in this series of athletes at increased risk of its elevation.

  14. Oral Supplementation with Bovine Colostrum Decreases Intestinal Permeability and Stool Concentrations of Zonulin in Athletes.

    Science.gov (United States)

    Hałasa, Maciej; Maciejewska, Dominika; Baśkiewicz-Hałasa, Magdalena; Machaliński, Bogusław; Safranow, Krzysztof; Stachowska, Ewa

    2017-04-08

    Increased intestinal permeability has been implicated in various pathologies, has various causes, and can develop during vigorous athletic training. Colostrum bovinum is a natural supplement with a wide range of supposed positive health effects, including reduction of intestine permeability. We assessed influence of colostrum supplementation on intestinal permeability related parameters in a group of 16 athletes during peak training for competition. This double-blind placebo-controlled study compared supplementation for 20 days with 500 mg of colostrum bovinum or placebo (whey). Gut permeability status was assayed by differential absorption of lactulose and mannitol (L/M test) and stool zonulin concentration. Baseline L/M tests found that six of the participants (75%) in the colostrum group had increased intestinal permeability. After supplementation, the test values were within the normal range and were significantly lower than at baseline. The colostrum group Δ values produced by comparing the post-intervention and baseline results were also significantly lower than the placebo group Δ values. The differences in stool zonulin concentration were smaller than those in the L/M test, but were significant when the Δ values due to intervention were compared between the colostrum group and the placebo group. Colostrum bovinum supplementation was safe and effective in decreasing of intestinal permeability in this series of athletes at increased risk of its elevation.

  15. Single-beam integrating sphere spectrophotometer for reflectance and transmittance measurements versus angle of incidence in the solar wavelength range on diffuse and specular samples

    Science.gov (United States)

    Nostell, Per; Roos, Arne; Rönnow, Daniel

    1999-05-01

    A multipurpose instrument for the measurement of reflectance and transmittance versus angle of incidence for both specular and diffuse samples in the solar wavelength range has been constructed and evaluated. The instrument operates in the single-beam mode and uses a common light source for three experimental setups. Two integrating spheres, 20 cm in diameter, are used for diffuse transmittance and reflectance measurements. The transmittance sphere can be turned around an axis through the sample to vary the angle of incidence. The reflectance sphere uses a center mounted sample and a special feature is the position of the detector, which is mounted on the sample holder at the center of the sphere. This way the detector always sees the same part of the sphere wall and no light can reach the detector directly from the sample. The third setup is an absolute instrument for specular samples. It uses a small averaging sphere as a detector. The detector is mounted on an arm which rotates around the center of the sample, and it can thus pick up both the reflected and transmitted beams including all multiply reflected components. The averaging sphere detector is insensitive to small side shifts of the detected beams and no multiple reflections between detector and optical system occur. In this report a number of calibration procedures are presented for the three experimental setups and models for the calculation of correct transmittance and reflectance values from measured data are presented. It is shown that for integrating sphere measurements, the geometry of the sphere and the diffusivity of the sample as well as the sphere wall reflectance and port losses are important factors that influence the result. For the center mounted configuration these factors are particularly important and special emphasis is given to the evaluation of the reflectance sphere model. All three instrument setups are calibrated using certified reference materials and nonscattering mirrors and

  16. The use of inulin-type fructans improves stool consistency in constipated children. A randomised clinical trial: pilot study.

    Science.gov (United States)

    Closa-Monasterolo, Ricardo; Ferré, Natalia; Castillejo-DeVillasante, Gemma; Luque, Veronica; Gispert-Llaurado, Mariona; Zaragoza-Jordana, Marta; Theis, Stephan; Escribano, Joaquin

    2017-08-01

    Constipation is a common disorder in children. The objective of this study is to assess the beneficial effects of a daily supplementation with Orafti ® inulin-type fructans in 2-5 year old constipated children. Double-blind, randomised, placebo-controlled parallel group trial where constipated children received two doses of 2 g Orafti ® inulin-type fructans (OF:IN) or placebo (maltodextrin) for 6 weeks. Primary outcome was stool consistency. Secondary outcomes were stool frequency and gastrointestinal symptoms. Twenty-two children were included, 17 completed the study protocol (nine and eight for the control and the OF:IN group, respectively). Results showed that Orafti ® inulin-type fructans supplemented children had softer stools (p = .003). The longitudinal analysis showed no significant changes in controls, whereas supplemented children increased their stool consistency from 2.2 to 2.6 on the modified Bristol scale for children (five items instead of seven) (p = .040). Prebiotic inulin-type fructans supplementation improves stool consistency in constipated 2-5-year old children. Clinicaltrials.gov, with number NCT02863848.

  17. High throughput and accurate serum proteome profiling by integrated sample preparation technology and single-run data independent mass spectrometry analysis.

    Science.gov (United States)

    Lin, Lin; Zheng, Jiaxin; Yu, Quan; Chen, Wendong; Xing, Jinchun; Chen, Chenxi; Tian, Ruijun

    2018-03-01

    Mass spectrometry (MS)-based serum proteome analysis is extremely challenging due to its high complexity and dynamic range of protein abundances. Developing high throughput and accurate serum proteomic profiling approach capable of analyzing large cohorts is urgently needed for biomarker discovery. Herein, we report a streamlined workflow for fast and accurate proteomic profiling from 1μL of blood serum. The workflow combined an integrated technique for highly sensitive and reproducible sample preparation and a new data-independent acquisition (DIA)-based MS method. Comparing with standard data dependent acquisition (DDA) approach, the optimized DIA method doubled the number of detected peptides and proteins with better reproducibility. Without protein immunodepletion and prefractionation, the single-run DIA analysis enables quantitative profiling of over 300 proteins with 50min gradient time. The quantified proteins span more than five orders of magnitude of abundance range and contain over 50 FDA-approved disease markers. The workflow allowed us to analyze 20 serum samples per day, with about 358 protein groups per sample being identified. A proof-of-concept study on renal cell carcinoma (RCC) serum samples confirmed the feasibility of the workflow for large scale serum proteomic profiling and disease-related biomarker discovery. Blood serum or plasma is the predominant specimen for clinical proteomic studies while the analysis is extremely challenging for its high complexity. Many efforts had been made in the past for serum proteomics for maximizing protein identifications, whereas few have been concerned with throughput and reproducibility. Here, we establish a rapid, robust and high reproducible DIA-based workflow for streamlined serum proteomic profiling from 1μL serum. The workflow doesn't need protein depletion and pre-fractionation, while still being able to detect disease-relevant proteins accurately. The workflow is promising in clinical application

  18. Synthesis, crystal structure and cytotoxic activity of ruthenium(II) piano-stool complex with N,N-chelating ligand

    Science.gov (United States)

    Rogala, Patrycja; Jabłońska-Wawrzycka, Agnieszka; Kazimierczuk, Katarzyna; Borek, Agnieszka; Błażejczyk, Agnieszka; Wietrzyk, Joanna; Barszcz, Barbara

    2016-12-01

    A mononuclear compound of the general formula [(η6-p-cymene)RuIICl(2,2‧-PyBIm)]PF6 has been synthesized from a bidentate N,N-donor ligand, viz. 2,-(2‧-pyridyl)benzimidazole (2,2‧-PyBIm) and the corresponding chloro-complex [(η6-p-cymene)Ru(μ-Cl)Cl]2 (precursor). The isolated coordination compound was characterized by IR, UV-vis and 1H, 13C NMR spectroscopies. The single crystal X-ray analysis of the complex reveals that the asymmetric part of the unit cell consists of two symmetrically independent, [(η6-p-cymene)RuCl(2,2‧-PyBIm)]+ cationic complexes. Each cation exhibits a pseudo-octahedral three-legged piano-stool geometry, in which three "legs" are occupied by one chloride ion and two nitrogen donor atoms of the chelating ligand 2,2‧-PyBIm. The Hirshfeld surface analysis of obtained complex was determined, too. The ionic nature of the compound is identified by a strong band at around 830 cm-1 due to the νP-F stretching mode of the PF6- counter ion. The electronic spectrum of this monomeric complex displays high intensity bands in the ultraviolet region assignable to π→π*/n→π* transitions, as well as a band attributable to the metal-to-ligand charge transfer (MLCT) dπ(Ru)→π*(L) transition. Additionally, the complex has been screened for its cytotoxicity against three human cancer lines: non-small cell lung carcinoma (A549), colon adenocarcinoma (HT29) and breast adenocarcinoma (MCF-7) as well as normal mice fibroblast cells (BALB/3T3). The complex demonstrated a moderate antiproliferative activity against the cell lines tested.

  19. Thermal transfer and apparent-dose distributions in poorly bleached mortar samples: Results from single grains and small aliquots of quartz

    DEFF Research Database (Denmark)

    Jain, M.; Thomsen, Kristina Jørkov; Bøtter-Jensen, L.

    2004-01-01

    -300 mum and compare the dose-distributions obtained from small aliquots and single-grain procedures. A comparison of three different methods viz. (a) first 5%, (b) probability plot and (c) comparison of internal and external uncertainties, is made for equivalent dose estimation. The results have......;, this process releases all the prior trapped charge and simultaneously sensitises the quartz. Unfortunately unheated materials such as mortar and concrete are more common in industrial sites and particularly in nuclear installations. These materials are usually exposed to daylight during quarrying...... and construction, but in general this exposure is insufficient to completely empty (bleach) any geological trapped charge. This leads to a distribution of apparent doses in the sample at the time of construction with only some (if any) grains exposed to sufficient light to be considered well bleached for OSL...

  20. Ultra-Trace Determination of Copper and Silver in Environmental Samples by Using Ionic Liquid-Based Single Drop Microextraction-Electrothermal Atomic Absorption Spectrometry

    Directory of Open Access Journals (Sweden)

    J. Abolhasani

    2013-11-01

    Full Text Available A sensitive, selective and effective ionic liquid-based single drop microextraction technique wasdeveloped by using ionic liquid, 1-hexyl-3-methylimidazolium hexafluorophosphate, C6MIMPF6, coupledwith electrothermal atomic absorption spectrometry (ETAAS for the determination of copper and silver inenvironmental samples. Dithizone was used as chelating agent. Several factors that influence themicroextraction efficiency and ETAAS signal, such as pH, dithizone concentration, extraction time, amounts ofionic liquid, stirring rate, pyrolysis and atomization temperature were investigated and the microextractionconditions were established. In the optimum experimental conditions, the detection limits (3 s of the methodwere 4 and 8 ng L-1 and corresponding relative standard deviations (0.1 μg L-1, n = 6 were 4.2% and 4.8% forAg and Cu, respectively. The developed method was validated by analysis of a certified reference material andapplied to the determination of silver and copper.

  1. Capillary electrophoresis coupled to evaporative light scattering detection for direct determination of underivatized amino acids: application to tea samples using carboxyled single-walled carbon nanotubes for sample preparation.

    Science.gov (United States)

    Bouri, Mohamed; Salghi, Rachid; Zougagh, Mohammed; Ríos, Angel

    2013-09-01

    An improved and efficient method for the determination of underivatized amino acids based on the use of CE coupled to evaporative light scattering detector (ELSD), involving carbon nanotubes, was successfully developed. Carboxyled single-walled carbon nanotubes were used for the first time to perform the clean-up of the analyzed samples, which were afterwards analyzed by CE-ELSD. White tea samples were used to demonstrate the usefulness of the CE-ELSD coupled methodology. A suitable interface, based on a triple tube design sprayer, was developed and successfully used for coupling both instruments. Parameters affecting the separation and determination, including the elimination of interferences, were studied and properly optimized. Under the optimized conditions good resolution was achieved for the separation of seven amino acids. The precision of the method, expressed as RSD, was found within the 3.5-5.3% range. The LOD obtained for the proposed method were in the 1.2-2.1 pg range and the LOQ, were in the 2.0-11.5 pg range, with injection pressure of 5 KPa for 20 s (15.3 nL). This method is simple, rapid, and selective compared with other conventional techniques. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Fluorescent dye imaging of the volume sampled by single well forced-gradient tracer tests evaluated in a laboratory-scale aquifer physical model.

    Science.gov (United States)

    Barns, Gareth L; Wilson, Ryan D; Thornton, Steven F

    2012-02-01

    This study presents a new method to visualise forced-gradient tracer tests in 2-D using a laboratory-scale aquifer physical model. Experiments were designed to investigate the volume of aquifer sampled in vertical dipole flow tracer tests (DFTT) and push-pull tests (PPT), using a miniature monitoring well and straddle packer arrangement equipped with solute injection and recovery chambers. These tests have previously been used to estimate bulk aquifer hydraulic and transport properties for the evaluation of natural attenuation and other remediation approaches. Experiments were performed in a silica glass bead-filled box, using a fluorescent tracer (fluorescein) to deduce conservative solute transport paths. Digital images of fluorescein transport were captured under ultraviolet light and processed to analyse tracer plume geometry and obtain point-concentration breakthrough histories. Inorganic anion mixtures were also used to obtain conventional tracer breakthrough histories. Concentration data from the conservative tracer breakthrough curves was compared with the digital images and a well characterised numerical model. The results show that the peak tracer breakthrough response in dipole flow tracer tests samples a zone of aquifer close to the well screen, while the sampling volume of push-pull tests is limited by the length of the straddle packers used. The effective sampling volume of these single well forced-gradient tests in isotropic conditions can be estimated with simple equations. The experimental approach offers the opportunity to evaluate under controlled conditions the theoretical basis, design and performance of DFTTs and PPTs in porous media in relation to measured flow and transport properties. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Integration of GC-MSD and ER-Calux® assay into a single protocol for determining steroid estrogens in environmental samples.

    Science.gov (United States)

    Avberšek, Miha; Žegura, Bojana; Filipič, Metka; Heath, Ester

    2011-11-01

    There are many published studies that use either chemical or biological methods to investigate steroid estrogens in the aquatic environment, but rarer are those that combine both. In this study, gas chromatography with mass selective detection (GC-MSD) and the ER-Calux(®) estrogenicity assay were integrated into a single protocol for simultaneous determination of natural (estrone--E1, 17β-estradiol--E2, estriol--E3) and synthetic (17α-ethinylestradiol--EE2) steroid estrogens concentrations and the total estrogenic potential of environmental samples. For integration purposes, several solvents were investigated and the commonly used dimethyl sulphoxide (DMSO) in the ER-Calux(®) assay was replaced by ethyl acetate, which is more compatible with gas chromatography and enables the same sample to be analysed by both GC-MSD and the ER-Calux(®) assay. The integrated protocol was initially tested using a standard mixture of estrogens. The results for pure standards showed that the estrogenicity calculated on the basis of GC-MSD and the ER-Calux(®) assay exhibited good correlation (r(2)=0.96; α=0.94). The result remained the same when spiked waste water extracts were tested (r(2)=0.92, α=1.02). When applied to real waste water influent and effluent samples the results proved (r(2)=0.93; α=0.99) the applicability of the protocol. The main advantages of this newly developed protocol are simple sample handling for both methods, and reduced material consumption and labour. In addition, it can be applied as either a complete or sequential analysis where the ER-Calux(®) assay is used as a pre-screening method prior to the chemical analysis. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Fluorescent dye imaging of the volume sampled by single well forced-gradient tracer tests evaluated in a laboratory-scale aquifer physical model

    Science.gov (United States)

    Barns, Gareth L.; Wilson, Ryan D.; Thornton, Steven F.

    2012-02-01

    This study presents a new method to visualise forced-gradient tracer tests in 2-D using a laboratory-scale aquifer physical model. Experiments were designed to investigate the volume of aquifer sampled in vertical dipole flow tracer tests (DFTT) and push-pull tests (PPT), using a miniature monitoring well and straddle packer arrangement equipped with solute injection and recovery chambers. These tests have previously been used to estimate bulk aquifer hydraulic and transport properties for the evaluation of natural attenuation and other remediation approaches. Experiments were performed in a silica glass bead-filled box, using a fluorescent tracer (fluorescein) to deduce conservative solute transport paths. Digital images of fluorescein transport were captured under ultraviolet light and processed to analyse tracer plume geometry and obtain point-concentration breakthrough histories. Inorganic anion mixtures were also used to obtain conventional tracer breakthrough histories. Concentration data from the conservative tracer breakthrough curves was compared with the digital images and a well characterised numerical model. The results show that the peak tracer breakthrough response in dipole flow tracer tests samples a zone of aquifer close to the well screen, while the sampling volume of push-pull tests is limited by the length of the straddle packers used. The effective sampling volume of these single well forced-gradient tests in isotropic conditions can be estimated with simple equations. The experimental approach offers the opportunity to evaluate under controlled conditions the theoretical basis, design and performance of DFTTs and PPTs in porous media in relation to measured flow and transport properties. DFTT = Dipole Flow Tracer Test, PPT = Push Pull Tracer Test.

  5. Amostragem por larva-única na vigilância de Aedes aegypti Single-larva sampling for Aedes aegypti surveillance

    Directory of Open Access Journals (Sweden)

    José Eduardo Bracco

    1995-04-01

    Full Text Available Com a finalidade de testar a metodologia de amostragem por larva-única na vigilância entomológica do Aedes aegypti, foram pesquisados domicílios do Município de Araraquara, SP (Brasil. Nos criadouros que continham larvas de Aedes uma delas foi coletada. Como controle, após a coleta da larva-única, todas as larvas foram coletadas para identificação posterior. Esse processo foi repetido no laboratório. Dos 447 domicílios visitados, apenas 12 foram considerados positivos e 20 criadouros foram identificados; destes, 13 continham larvas de Aedes; 5, larvas de Aedes e Culex e 2, larvas de Culex. Os resultados mostram o reconhecimento correto, no campo, de todos os criadouros, evidenciando que o método poderia ser utilizado na vigilância entomológica de municípios sem infestação domiciliar ou infestados apenas com uma única espécie de Aedes.Buildings in Araraquara city, Southeastern Brazil, were searched during a year for the presence of Aedes larvae using single larva sampling in order to check the single-larva methodology. In those breeding places in wich Aedes larvae were found, one of them was collected. As a control, after the single larva had been collected, all the larvae from the breeding place were collected for later identification. This process was repeated in the laboratory. Of the 447 domiciles searched, 12 were considered positive and 20 breeding places were found. Of the breeding places, 13 contained Aedes larvae, 5 both Aedes and Culex larvae and 2 Culex larvae only. The results show that all the breeding places in the field were properly recognited showing the method may be used for Aedes surveillance in cities infested with one species only or without any domiciliary infestation.

  6. The structural and magnetic properties of CsxFe2-ySe2 as determined by x ray and neutron scattering of powder and single crystal samples

    Science.gov (United States)

    Taddei, Keith; Chmaissem, Omar; Sturza, Mihai; Avci, Sevda; Claus, Helmut; Kanatzids, Mercouri; Rosenkranz, Stephan; Osborn, Ray

    2014-03-01

    The AxFe2-ySe2 family of iron selenides (A = K, Rb and Cs) has proven an intricate system for the study of unconventional superconductivity, exhibiting high temperature superconductivity (~ 30 K) and a complex structural phase transition into a biphasic state coupled with a high temperature magnetic transition (~ 500 K). While isostructural to the 122 arsenides, significant structural differences are identified. In the selenides, iron vacancies in the tetrahedral FeSe layers become ordered below a high temperature structural transition defining a main phase √ 5 × √ 5 superstructure. Coexistent with the main phase, a secondary phase of a previously contested structure is observed and it is in this biphasic state that superconductivity arises at ~ 30 K. Both powder and single crystal samples show similar phase separation and coexistence. In this talk, I will discuss structural results and lattice parameter evolution obtained from neutron powder diffraction as well as single crystal x-ray diffraction with an emphasis on a novel magnetic structural model, the identification of the secondary phase, and the nature of coincidence of the magnetic, structural and secondary phase transitions.

  7. Determination of {sup 236}U in environmental samples by single extraction chromatography coupled to triple-quadrupole inductively coupled plasma-mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Guosheng [Department of Radiation Chemistry, Institute of Radiation Emergency Medicine, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, 036-8564 (Japan); Division of Nuclear Technology and Applications, Institute of High Energy Physics, Chinese Academy of Sciences (China); Beijing Engineering Research Center of Radiographic Techniques and Equipment, Beijing, 100049 (China); Tazoe, Hirofumi [Department of Radiation Chemistry, Institute of Radiation Emergency Medicine, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, 036-8564 (Japan); Yamada, Masatoshi, E-mail: myamada@hirosaki-u.ac.jp [Department of Radiation Chemistry, Institute of Radiation Emergency Medicine, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, 036-8564 (Japan)

    2016-11-09

    In order to measure trace {sup 236}U and {sup 236}U/{sup 238}U in environmental samples with a high matrix effect, a novel and simple method was developed that makes the digestion and purification procedures compatible with advanced triple-quadrupole inductively coupled plasma-mass spectrometry. A total dissolution of sample with HF + HNO{sub 3} + HClO{sub 4} was followed by chromatographic separation with a single resin column containing normal type DGA resin (N,N,N′,N’-tetra-n-octyldiglycolamide) as the extractant system. The analytical accuracy and precision of {sup 236}U/{sup 238}U ratios, measured as {sup 236}U{sup 16}O{sup +}/{sup 238}U{sup 16}O{sup +}, were examined by using the reference materials IAEA-135, IAEA-385, IAEA-447, and JSAC 0471. The low method detection limit (3.50 × 10{sup −6} Bq kg{sup −1}) makes it possible to perform routine monitoring of environmental {sup 236}U due to global fallout combined with the Fukushima Daiichi Nuclear Power Plant accident fallout (>10{sup −5} Bq kg{sup −1}). Finally, the developed method was successfully applied to measure {sup 236}U/{sup 238}U ratios and {sup 236}U activities in soil samples contaminated by the accident. The low {sup 236}U/{sup 238}U atom ratios ((1.50–13.5) × 10{sup −8}) and {sup 236}U activities ((2.25–14.1) × 10{sup −2} mBq kg{sup −1}) indicate {sup 236}U contamination was mainly derived from global fallout in the examined samples. - Highlights: • A simple {sup 236}U/{sup 238}U analytical method has been developed. • The separation required just one DGA column chromatography. • {sup 236}U/{sup 238}U atom ratios in soil were measured by ICP-MS/MS. • {sup 236}U/{sup 238}U atom ratios of (1.50–13.5) × 10{sup −8} were observed in Japanese samples. • {sup 236}U activities of (2.25–14.1) × 10{sup −2} mBq kg{sup −1} were found in Japanese samples.

  8. Photothermal desorption of single-walled carbon nanotubes and coconut shell-activated carbons using a continuous light source for application in air sampling.

    Science.gov (United States)

    Floyd, Evan L; Sapag, Karim; Oh, Jonghwa; Lungu, Claudiu T

    2014-08-01

    Many techniques exist to measure airborne volatile organic compounds (VOCs), each with differing advantages; sorbent sampling is compact, versatile, has good sample stability, and is the preferred technique for collecting VOCs for hygienists. Development of a desorption technique that allows multiple analyses per sample (similar to chemical desorption) with enhanced sensitivity (similar to thermal desorption) would be helpful to field hygienists. In this study, activated carbon (AC) and single-walled carbon nanotubes (SWNT) were preloaded with toluene vapor and partially desorbed with light using a common 12-V DC, 50-W incandescent/halogen lamp. A series of experimental chamber configurations were explored starting with a 500-ml chamber under static conditions, then with low ventilation and high ventilation, finally a 75-ml high ventilation chamber was evaluated. When preloaded with toluene and irradiated at the highest lamp setting for 4min, AC desorbed 13.9, 18.5, 23.8, and 45.9% of the loaded VOC mass, in each chamber configuration, respectively; SWNT desorbed 25.2, 24.3, 37.4, and 70.5% of the loaded VOC mass, respectively. SWNT desorption was significantly greater than AC in all test conditions (P = 0.02-<0.0001) demonstrating a substantial difference in sorbent performance. When loaded with 0.435mg toluene and desorbed at the highest lamp setting for 4min in the final chamber design, the mean desorption for AC was 45.8% (39.7, 52.0) and SWNT was 72.6% (68.8, 76.4) (mean represented in terms of 95% confidence interval). All desorption measurements were obtained using a field grade photoionization detector; this demonstrates the potential of using this technique to perform infield prescreening of VOC samples for immediate exposure feedback and in the analytical lab to introduce sample to a gas chromatograph for detailed analysis of the sample. © The Author 2014. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.

  9. Detection of non-jejuni and -coli Campylobacter species from stool specimens with an immunochromatographic antigen detection assay.

    Science.gov (United States)

    Couturier, Brianne A; Couturier, Marc Roger; Kalp, Kim J; Fisher, Mark A

    2013-06-01

    The STAT! Campy immunochromatographic assay for Campylobacter antigen was compared to culture for 500 clinical stool specimens. Antigen was detected in six culture-negative, PCR-positive specimens. C. upsaliensis, a pathogenic species that is traditionally difficult to recover in routine stool cultures, was detected in two of these culture-negative specimens. This study provides evidence that antigen testing may cross-react with at least one additional non-jejuni and -coli Campylobacter species that may be missed by routine culture for campylobacteriosis.

  10. Antibacterial Screening on Leaves of Argyreia Cymosa Roxb. Against Pathogenic Bacteria Isolated from Infected Pateints Samples Wound, Sputum and Stool

    OpenAIRE

    N. Packialakshmi; H. Fazila Beevi

    2014-01-01

    The present study deals with the aqueous leaf extract and synthesized silver nanoparticle of Argyreia cymosa (Roxb) were evaluated for antibacterial activity. The aqueous leaf extract and synthesized nanoparticle of Argyreia cymosa is active against E. coli, P. aeruginosa, Bacillus and S. aureus. The aqueous leaf extract showed maximum activity against Bacillus (20mm), S.aureus (18mm), P. aeruginosa (13mm) E.coli (12mm) and the synthesized silver nano particle showed maximum activity against ...

  11. Whole-genome sequencing of Campylobacter jejuni isolated from Danish routine human stool samples reveals surprising degree of clustering

    DEFF Research Database (Denmark)

    Joensen, K G; Kuhn, K G; Müller, L

    2018-01-01

    OBJECTIVES: Outbreaks of Campylobacter are traditionally considered to be rare, however rather than being the true nature of the disease, this may reflect our present inability to detect them. The aim of this study was to determine the genetic and epidemiological degree of clustering among Campyl...

  12. Diagnostic Algorithm Using a Sensitive Broth Culture Method for Detection of Clostridium difficile Toxin from Stool Samples

    Directory of Open Access Journals (Sweden)

    Paul Bayardelle

    2009-01-01

    Full Text Available BACKGROUND: The two-step glutamate dehydrogenase antigencytotoxicity neutralization assay algorithm has been found to be reliable for the diagnosis of toxigenic Clostridium difficile. However, the high sensitivity of the screening method is compromised by the relative low sensitivity of the second step, the direct cytotoxin neutralization assay (DCNA using a fecal filtrate. The objective of the present study was to compare the DCNA with an indirect cytotoxin neutralization assay (ICNA.

  13. The Bristol Stool Form Scale: its translation to Portuguese, cultural adaptation and validation.

    Science.gov (United States)

    Martinez, Anna Paula; de Azevedo, Gisele Regina

    2012-01-01

    The Bristol Stool Form Scale is used for describing feces. The objective of this research was its translation, cultural adaptation and validation for Brazil. The methodology was translation, back-translation and discussion. Validation involved 85 nurses, 80 doctors, and 80 patients, who correlated images of seven types of feces with the descriptions. there was a difference in sex distribution, with males predominating among the doctors and females among nurses and patients. In relation to concordance between definitions and pictures, the highest percentage was in type 5 in all three groups and the lowest was in types 6 and 7 for the doctors, in type 3 for the nurses, and type 6 for the patients. The general Kappa index was 0.826. the scale demonstrated high reliability for all the groups studied.

  14. A Comparative Study of Single-pulse and Double-pulse Laser-Induced Breakdown Spectroscopy with Uranium-containing Samples.

    Science.gov (United States)

    Skrodzki, Patrick J; Becker, Jason R; Diwakar, Prasoon K; Harilal, Sivanandan S; Hassanein, Ahmed

    2016-03-01

    Laser-induced breakdown spectroscopy (LIBS) holds potential advantages in special nuclear material (SNM) sensing and nuclear forensics, which require rapid analysis, minimal sample preparation, and stand-off distance capability. SNM, such as U, however, result in crowded emission spectra with LIBS, and characteristic emission lines are challenging to discern. It is well-known that double-pulse LIBS (DPLIBS) improves the signal intensity for analytes over conventional single-pulse LIBS (SPLIBS). This study investigates the U signal in a glass matrix using DPLIBS and compares it to signal obtained using SPLIBS. Double-pulse LIBS involves sequential firing of a 1.06 µm Nd:YAG pre-pulse and 10.6 µm TEA CO2 heating pulse in a near collinear geometry. Optimization of experimental parameters including inter-pulse delay and energy follows identification of characteristic lines for the bulk analyte Ca and the minor constituent analyte U for both DPLIBS and SPLIBS. Spatial and temporal coupling of the two pulses in the proposed DPLIBS technique yields improvements in analytical merits with a negligible increase in damage to the sample compared to SPLIBS. Subsequently, the study discusses optimum plasma emission conditions of U lines and relative figures of merit in both SPLIBS and DPLIBS. Investigation into plasma characteristics also addresses plausible mechanisms related to the observed U analyte signal variation between SPLIBS and DPLIBS. © The Author(s) 2016.

  15. Helicobacter pylori from Peptic Ulcer Patients in Uganda Is Highly Resistant to Clarithromycin and Fluoroquinolones: Results of the GenoType HelicoDR Test Directly Applied on Stool

    Directory of Open Access Journals (Sweden)

    Denish Calmax Angol

    2017-01-01

    Full Text Available Background. Around 70–90% of peptic ulcer disease (PUD is due to Helicobacter pylori and requires treatment with antimicrobials to which these bacteria are susceptible. Common H. pylori diagnostic tests do not provide drug susceptibility data. Using the GenoType HelicoDR PCR test designed for gastric biopsies for simultaneous detection of H. pylori and its resistance to clarithromycin (CLA/fluoroquinolones (FLQ, we present evidence for stool as an optional test specimen and also provide data on prevalence of H. pylori resistance to CLA and FLQ in Uganda. Methods. Stool from 142 symptomatic PUD patients at three hospitals in Kampala was screened for H. pylori using a rapid antigen test. The GenoType HelicoDR test was run on all H. pylori antigen positives to determine PCR positivity and resistance to CLA/FLQ. Results. Thirty-one samples (22% were H. pylori antigen positive, and 21 (68% of these were H. pylori PCR positive. Six of the 21 (29% were resistant to CLA and eight to FLQ (42%, while two gave invalid FLQ resistance results. Conclusion. Stool is a possible specimen for the GenoType HelicoDR test for rapid detection of H. pylori and drug resistance. In Uganda, Helicobacter pylori is highly resistant to CLA and FLQ.

  16. Morphologic changes of the anal sphincter musculature during and after temporary stool deviation.

    Science.gov (United States)

    Sailer, M; Fein, M; Fuchs, K H; Bussen, D; Grun, C; Thiede, A

    2001-04-01

    Temporary stool deviation, using a stoma, is a well-known surgical principle to protect low colorectal or coloanal anastomoses. The purpose of this study was to evaluate any morphologic changes with regard to the anal sphincter muscles during and after temporary ileostomy. Forty-four patients with rectal carcinomas were studied prospectively. All patients underwent low anterior resection. Reconstruction was performed using either a coloanal pouch or a straight end-to-end anastomosis. A protective stoma was fashioned in all 44 patients (ileostomy n=41; colostomy n=3). Stoma closure was carried out after a median of 85 days (41-330 days). Using a standard protocol, anal-sphincter thickness [m. puborectalis, external anal sphincter (EAS) and internal anal (IAS) sphincter] was assessed by means of endoanal ultrasonography preoperatively, at the time of stoma closure, and every 3 months thereafter for 1 year. The diameter of the puborectal muscle decreased from a median preoperative value of 6.3 mm to 5.7 mm at the time of stoma closure (P=0.03). After 3 months, 6.2 mm was measured. This value remained stable for the complete follow-up period. Similar results were recorded for the EAS. The IAS thickness remained stable throughout the study period, measuring between 2.1 mm and 2.4 mm. Temporary stool deviation does lead to morphologic changes of the anal sphincter. While the smooth muscle remains unchanged, the striated counterpart undergoes atrophic transformation. However, after passage reconstruction, i.e., stoma closure, a rapid regeneration of the voluntary muscles is observed.

  17. Environmental Stress Testing of the Single Sample Cylinder: A Proven Consensus Standard for Internal Gas Analysis (IGA) or Residual Gas Analysis (RGA)

    Science.gov (United States)

    Schuessler, Philipp WH

    2010-01-01

    In August 2008, Schuessler Consulting was contracted by NASA GSFC in support of the NASA Electronic Parts and Packaging (NEPP) program to perform two separate studies on moisture laden air in a stainless steel cylinder that had been designed to become a consensus standard for Test Method 1018. This Test Method was originally released for hybrids under Mil. Std. 883 but was quickly utilized on other microelectronic devices under the auspice of Mil. Std. 750. The cylinder had subsequently been fabricated for the 750 community. It was back-filled with moist air and subsequently analyzed over a period of time under a previous NASA contract. It had been shown that moisture in the 4000 - 5000 ppm range could be analyzed rather precisely with a mass spectrometer, commonly referred to as a Residual Gas Analyzer (RGA). The scope of this study was to ascertain if the composition and precision varied as a function of thermal shock at sub-zero temperatures and whether there was consensus when the standard was submitted to other RGA units. It was demonstrated and published that the consensus standard would yield precise RGA data for moisture within +/- 1% when optimized for a given RGA unit. It has been subsequently shown in this study at Oneida Research Services, that sub-zero storage did not affect that precision when a well-defined protocol for the analysis was followed. The consensus standard was taken to a second facility for analysis where it was found that moisture adsorption on the transfer lines caused precision to drop to +/- 12%. The Single Sample Cylinder (SSC) is a one liter stainless steel cylinder with associated sampling valves and has considerable weight and volume. But this considerable size allows for approximately 300 gas samples of the same composition to be delivered to any RGA unit. Lastly, a smaller cylinder, approximately 75 cc, of a second consensus standard was fabricated and tested with a different mix of fixed gases where moisture was kept in the

  18. Estimating the effective number of breeders from single parr samples for conservation monitoring of wild populations of Atlantic salmon Salmo salar.

    Science.gov (United States)

    Bacles, C F E; Bouchard, C; Lange, F; Manicki, A; Tentelier, C; Lepais, O

    2018-03-01

    This study assesses whether the effective number of breeders (N b ) can be estimated using a time and cost-effective protocol using genetic sibship reconstruction from a single sample of young-of-the-year (YOY) for the purposes of Atlantic salmon Salmo salar population monitoring. N b was estimated for 10 consecutive reproductive seasons for S. salar in the River Nivelle, a small population located at the rear-edge of the species distribution area in France, chronically under its conservation limit and subjected to anthropogenic and environmental changes. Subsampling of real and simulated data showed that accurate estimates of N b can be obtained from YOY genotypes, collected at moderate random sampling intensity, achievable using routine juvenile electrofishing protocols. Spatial bias and time elapsed since spawning were found to affect estimates, which must be accounted for in sampling designs. N b estimated in autumn for S. salar in the River Nivelle was low and variable across years from 23 (95% C.I. 14-41) to 75 (53-101) and was not statistically correlated with the estimated number of returning adults, but it was positively correlated with the estimated number of YOY at age 9 months. N b was found to be lower for intermediate levels of redd aggregation, suggesting that the strength of the competition between males to access females affects reproductive success variance depending on redd spatial configuration. Thus, environmental factors such as habitat availability and quality for spawning and YOY development predominate over demographic ones (number of returning adults) in driving long-term population viability for S. salar in the River Nivelle. This study showcases N b as an integrated parameter, encompassing demographic and ecological information about a reproductive event, relevant to the assessment of both short-term effects of management practices and long-term population conservation status. © 2018 The Fisheries Society of the British Isles.

  19. Spectroelectrochemical Sensing Based on Multimode Selectivity simultaneously Achievable in a Single Device. 11. Design and Evaluation of a Small Portable Sensor for the Determination of Ferrocyanide in Hanford Waste Samples

    International Nuclear Information System (INIS)

    Stegemiller, Michael L.; Heineman, William R.; Seliskar, Carl J.; Ridgway, Thomas H.; Bryan, Samuel A.; Hubler, Timothy L.; Sell, Richard L.

    2003-01-01

    Spectroelectrochemical sensing based on multimode selectivity simultaneously achievable in a single device. 11. Design and evaluation of a small portable sensor for the determination of ferrocyanide in Hanford waste samples

  20. ‘N-of-1-pathways’ unveils personal deregulated mechanisms from a single pair of RNA-Seq samples: towards precision medicine

    Science.gov (United States)

    Gardeux, Vincent; Achour, Ikbel; Li, Jianrong; Maienschein-Cline, Mark; Li, Haiquan; Pesce, Lorenzo; Parinandi, Gurunadh; Bahroos, Neil; Winn, Robert; Foster, Ian; Garcia, Joe G N; Lussier, Yves A

    2014-01-01

    Background The emergence of precision medicine allowed the incorporation of individual molecular data into patient care. Indeed, DNA sequencing predicts somatic mutations in individual patients. However, these genetic features overlook dynamic epigenetic and phenotypic response to therapy. Meanwhile, accurate personal transcriptome interpretation remains an unmet challenge. Further, N-of-1 (single-subject) efficacy trials are increasingly pursued, but are underpowered for molecular marker discovery. Method ‘N-of-1-pathways’ is a global framework relying on three principles: (i) the statistical universe is a single patient; (ii) significance is derived from geneset/biomodules powered by paired samples from the same patient; and (iii) similarity between genesets/biomodules assesses commonality and differences, within-study and cross-studies. Thus, patient gene-level profiles are transformed into deregulated pathways. From RNA-Seq of 55 lung adenocarcinoma patients, N-of-1-pathways predicts the deregulated pathways of each patient. Results Cross-patient N-of-1-pathways obtains comparable results with conventional genesets enrichment analysis (GSEA) and differentially expressed gene (DEG) enrichment, validated in three external evaluations. Moreover, heatmap and star plots highlight both individual and shared mechanisms ranging from molecular to organ-systems levels (eg, DNA repair, signaling, immune response). Patients were ranked based on the similarity of their deregulated mechanisms to those of an independent gold standard, generating unsupervised clusters of diametric extreme survival phenotypes (p=0.03). Conclusions The N-of-1-pathways framework provides a robust statistical and relevant biological interpretation of individual disease-free survival that is often overlooked in conventional cross-patient studies. It enables mechanism-level classifiers with smaller cohorts as well as N-of-1 studies. Software http://lussierlab.org/publications/N-of-1-pathways

  1. Fully automated ionic liquid-based headspace single drop microextraction coupled to GC-MS/MS to determine musk fragrances in environmental water samples.

    Science.gov (United States)

    Vallecillos, Laura; Pocurull, Eva; Borrull, Francesc

    2012-09-15

    A fully automated ionic liquid-based headspace single drop microextraction (IL-HS-SDME) procedure has been developed for the first time to preconcentrate trace amounts of ten musk fragrances extensively used in personal care products (six polycyclic musks, three nitro musks and one polycyclic musk degradation product) from wastewater samples prior to analysis by gas chromatography and ion trap tandem mass spectrometry (GC-IT-MS/MS). Due to the low volatility of the ILs, a large internal diameter liner (3.4 mm i.d.) was used to improve the ILs evaporation. Furthermore, a piece of glass wool was introduced into the liner to avoid the entrance of the ILs in the GC column and a guard column was used to prevent analytical column damages. The main factors influencing the IL-HS-SDME were optimized. For all species, the highest enrichments factors were achieved using 1 μL of 1-octyl-3-methylimidazolium hexafluorophosphate ([OMIM][PF(6)]) ionic liquid exposed in the headspace of 10 mL water samples containing 300 g L(-1) of NaCl and stirred at 750 rpm and 60 °C for 45 min. All compounds were determined by direct injection GC-IT-MS/MS with a chromatographic time of 19 min. Method detection limits were found in the low ng mL(-1) range between 0.010 ng mL(-1) and 0.030 ng mL(-1) depending on the target analytes. Also, under optimized conditions, the method gave good levels of intra-day and inter-day repeatabilities in wastewater samples with relative standard deviations varying between 3% and 6% and 5% and 11%, respectively (n=3, 1 ng mL(-1)). The applicability of the method was tested with different wastewater samples from influent and effluent urban wastewater treatment plants (WWTPs) and one potable treatment plant (PTP). The analysis of influent urban wastewater revealed the presence of galaxolide and tonalide at concentrations of between 2.10 ng mL(-1) and 0.29 ng mL(-1) and 0.32 ng mL(-1) and MQL (Method Quantification Limit), respectively; while the remaining

  2. Collective Ion-Pair Single-Drop Microextraction Attenuated Total Reflectance Fourier Transform Infrared Spectroscopic Determination of Perchlorate in Bioenvironmental Samples.

    Science.gov (United States)

    Chandrawanshi, Swati; Verma, Santosh K; Deb, Manas K

    2017-09-28

    Perchlorate (ClO₄ - ) is an environmental pollutant that affects human health. Perchlorate acts as a competitive inhibitor of iodine uptake in the thyroid gland (sodium-iodide symporter inhibitor); thus, its determination is important for public health concerns. Water and milk constitute a significant portion of the human diet. Because regular intake leads to an increase in perchlorate concentration in the human body, the estimation of perchlorate is of great concern. In this work, ion-pair single-drop microextraction (SDME) combined with attenuated total reflectance (ATR)-FTIR spectroscopy has been developed for the determination of perchlorate in bioenvironmental (soil, water, dairy milk, breast milk, and urine) samples. Perchlorate was extracted in a single drop of methyl isobutyl ketone as an - with the cationic surfactant cetyltrimethylammonuim bromide under optimized conditions. The strongest IR peak (at 1076 cm -1 ) was selected for the quantification of perchlorate among three observed vibrational peaks. Eight calibration curves for different concentration ranges of perchlorate were prepared, and excellent linearity was observed for absorbance and peak area in the range of 0.03-100 ng/mL perchlorate, with r values of 0.977 and 0.976, respectively. The RSDs ( n = 8) for the perchlorate concentration ranges of 0.03-100, 0.03-0.5, 0.5-10, and 10-100 ng/mL were in the range of 1.9-2.7% for the above calibration curves. The LOD and LOQ in the present work were 0.003 and 0.02 ng/mL, respectively. The extracted microdrop was analyzed directly by ATR-FTIR spectroscopy. The parameters affecting SDME, i.e, effect of pH, stirring rate, reagent concentration, microdrop volume, and extraction time, were optimized, and the role of foreign species was also investigated. F - and t -tests were performed to check the analytical QA of the method. A noteworthy feature of the reported method is the noninterference of any of the associated ions. The results were compared with

  3. The accuracy of the Helicobacter pylori stool antigen test in diagnosing H-pylori in treated and untreated patients

    NARCIS (Netherlands)

    Arents, NL; van Zwet, AA; Thijs, JC; de Jong, A; Pool, MO; Kleibeuker, JH

    Objective and design To evaluate the performance of the Helicobacter pylori stool antigen test (HpSA test) in detecting H. pylori infection and monitoring the effect of treatment. This was done in two separate studies using either a biopsy or the C-13-urea breath test based 'gold standard' (in

  4. Evaluation of stool microbiota signatures in two cohorts of Asian (Singapore and Indonesia newborns at risk of atopy

    Directory of Open Access Journals (Sweden)

    Chua Kaw Yan

    2011-08-01

    Full Text Available Abstract Background Studies have suggested that demographic and lifestyle factors could shape the composition of fecal microbiota in early life. This study evaluated infant stool microbiota signatures in two Asian populations, Singapore (n = 42 and Indonesia (n = 32 with contrasting socioeconomic development, and examined the putative influences of demographic factors on these human fecal associated bacterial signatures. Results Longitudinal analysis showed associations of geographical origin with Clostridium leptum, Atopobium and Bifidobacterium groups. Mode of delivery had the largest effect on stool microbiota signatures influencing the abundance of four bacterial groups. Significantly higher abundance of bacterial members belonging to the Bacteroides-Prevotella, Bifidobacterium and Atopobium groups, but lower abundance of Lactobacilli-Enterococci group members, were observed in vaginal delivered compared to caesarean delivered infants. Demographic factors influencing the structure of infants stool microbiota during the first year of life included breastfeeding, age of weaning, sibship size and exposure to antibiotics. Conclusions Differences in stool microbiota signatures were observed in relation to various demographic factors. These features may confound studies relating to the association of the structure of fecal microbiota and the predisposition to human modern disease.

  5. Helicobacter pylori Stool Antigen test: a reliable non-invasive test for the diagnosis of Helicobacter pylori infection in children

    NARCIS (Netherlands)

    van Doorn, O. J.; Bosman, D. K.; van't Hoff, B. W.; Taminiau, J. A.; ten Kate, F. J.; van der Ende, A.

    2001-01-01

    OBJECTIVE: To evaluate the Helicobacter pylori Stool Antigen (HpSA) test for the diagnosis of H. pylori infection in children. DESIGN AND SETTING: Prospective cohort study in an academic medical centre. PATIENTS AND METHODS: A total of 106 consecutive children who underwent gastroscopy were

  6. Effective testing for pulmonary tuberculosis using Xpert MTB/RIF assay for stool specimens in immunocompetent Pakistani children

    Directory of Open Access Journals (Sweden)

    Zahra Hasan

    2016-01-01

    Conclusion: Use of Xpert MTB/RIF assay for stool-based diagnosis of pulmonary TB in immunocompetent children is useful in a resource poor setting. This is a valuable and noninvasive diagnostic alternative for the diagnosis of childhood TB and can be adapted by pediatric arms of national TB programs.

  7. Highly sensitive stool DNA testing of Fusobacterium nucleatum as a marker for detection of colorectal tumours in a Japanese population.

    Science.gov (United States)

    Suehiro, Yutaka; Sakai, Kouhei; Nishioka, Mitsuaki; Hashimoto, Shinichi; Takami, Taro; Higaki, Shingo; Shindo, Yoshitaro; Hazama, Shoichi; Oka, Masaaki; Nagano, Hiroaki; Sakaida, Isao; Yamasaki, Takahiro

    2017-01-01

    Background Accumulating evidence shows an over-abundance of Fusobacterium nucleatum in colorectal tumour tissues. Although stool DNA testing of Fusobacterium nucleatum might be a potential marker for the detection of colorectal tumours, the difficulty in detecting Fusobacterium nucleatum in stool by conventional methods prevented further explorations. Therefore, we developed a droplet digital polymerase chain reaction (PCR) assay for detecting Fusobacterium nucleatum in stool and investigated its clinical utility in the management of colorectal tumours in a Japanese population. Methods Feces were collected from 60 healthy subjects (control group) and from 11 patients with colorectal non-advanced adenomas (non-advanced adenoma group), 19 patients with colorectal advanced adenoma/carcinoma in situ (advanced adenoma/carcinoma in situ (CIS) group) and 158 patients with colorectal cancer of stages I to IV (colorectal cancer group). Absolute copy numbers of Fusobacterium nucleatum were measured by droplet digital PCR. Results The median copy number of Fusobacterium nucleatum was 17.5 in the control group, 311 in the non-advanced adenoma group, 122 in the advanced adenoma/CIS group, and 317 in the colorectal cancer group. In comparison with that in the control group, the Fusobacterium nucleatum level was significantly higher in the non-advanced adenoma group, the advanced adenoma/CIS group and the colorectal cancer group. Conclusions This study illustrates the potential of stool DNA testing of Fusobacterium nucleatum by droplet digital PCR to detect individuals with colorectal tumours in a Japanese population.

  8. High transport efficiency of nanoparticles through a total-consumption sample introduction system and its beneficial application for particle size evaluation in single-particle ICP-MS.

    Science.gov (United States)

    Miyashita, Shin-Ichi; Mitsuhashi, Hiroaki; Fujii, Shin-Ichiro; Takatsu, Akiko; Inagaki, Kazumi; Fujimoto, Toshiyuki

    2017-02-01

    In order to facilitate reliable and efficient determination of both the particle number concentration (PNC) and the size of nanoparticles (NPs) by single-particle ICP-MS (spICP-MS) without the need to correct for the particle transport efficiency (TE, a possible source of bias in the results), a total-consumption sample introduction system consisting of a large-bore, high-performance concentric nebulizer and a small-volume on-axis cylinder chamber was utilized. Such a system potentially permits a particle TE of 100 %, meaning that there is no need to include a particle TE correction when calculating the PNC and the NP size. When the particle TE through the sample introduction system was evaluated by comparing the frequency of sharp transient signals from the NPs in a measured NP standard of precisely known PNC to the particle frequency for a measured NP suspension, the TE for platinum NPs with a nominal diameter of 70 nm was found to be very high (i.e., 93 %), and showed satisfactory repeatability (relative standard deviation of 1.0 % for four consecutive measurements). These results indicated that employing this total consumption system allows the particle TE correction to be ignored when calculating the PNC. When the particle size was determined using a solution-standard-based calibration approach without an NP standard, the particle diameters of platinum and silver NPs with nominal diameters of 30-100 nm were found to agree well with the particle diameters determined by transmission electron microscopy, regardless of whether a correction was performed for the particle TE. Thus, applying the proposed system enables NP size to be accurately evaluated using a solution-standard-based calibration approach without the need to correct for the particle TE.

  9. Sensitive and rapid detection of campylobacter species from stools of children with diarrhea in Japan by the loop-mediated isothermal amplification method.

    Science.gov (United States)

    Ushijima, Hiroshi; Nishimura, Shuichi; Thongprachum, Aksara; Shimizu-Onda, Yuko; Tran, Dinh Nguyen; Pham, Ngan Thi Kim; Takanashi, Sayaka; Dey, Shuvra Kanti; Okitsu, Shoko; Yamazaki, Wataru; Mizuguchi, Masashi; Hayakawa, Satoshi

    2014-01-01

    We detected Campylobacter spp. in 5% (20/380) of diarrheal stool samples collected at an outpatient clinic in Kyoto using a commercial loop-mediated isothermal amplification (LAMP) kit with a fluorescent detection reagent after DNA extraction. The sensitivity and specificity were 100% in comparison with those of semi-nested PCR for the differentiation of Campylobacter jejuni and Campylobacter coli. Fourteen of the 20 samples were already determined as C. jejuni by the culture method. All 20 samples were also positive for C. jejuni by the PCR method. Among the 58 cultured samples, the sensitivity of the culture method against the LAMP method was 93.3% (14/15) and the specificity was 100% (43/43). The detection rate of Campylobacter spp. from the heated supernatants by the LAMP method was lower than that from the supernatant after DNA extraction. In total, 25% (5/20) of the Campylobacter-positive samples by the LAMP method were co-infected with norovirus (3/20), rotavirus (1/20), and human parechovirus (1/20), although no other bacterial co-infection was identified by the culture method. C. jejuni was mostly detected in children aged >5 years throughout the year. Based on these results, we concluded that care should be taken while diagnosing Campylobacter infection in children. Our newly modified LAMP method is a rapid, easy, and useful method for this diagnosis.

  10. Feasibility of a Humor Training to Promote Humor and Decrease Stress in a Subclinical Sample: A Single-Arm Pilot Study

    Directory of Open Access Journals (Sweden)

    Nektaria Tagalidou

    2018-04-01

    Full Text Available The present study investigates the feasibility of a humor training for a subclinical sample suffering from increased stress, depressiveness, or anxiety. Based on diagnostic interviews, 35 people were invited to participate in a 7-week humor training. Evaluation measures were filled in prior training, after training, and at a 1-month follow-up including humor related outcomes (coping humor and cheerfulness and mental health-related outcomes (perceived stress, depressiveness, anxiety, and well-being. Outcomes were analyzed using repeated-measures ANOVAs. Within-group comparisons of intention-to-treat analysis showed main effects of time with large effect sizes on all outcomes. Post hoc tests showed medium to large effect sizes on all outcomes from pre to post and results remained stable until follow-up. Satisfaction with the training was high, attrition rate low (17.1%, and participants would highly recommend the training. Summarizing the results, the pilot study showed promising effects for people suffering from subclinical symptoms. All outcomes were positively influenced and showed stability over time. Humor trainings could be integrated more into mental health care as an innovative program to reduce stress whilst promoting also positive emotions. However, as this study was a single-arm pilot study, further research (including also randomized controlled trials is still needed to evaluate the effects more profoundly.

  11. Applying meta-pathway analyses through metagenomics to identify the functional properties of the major bacterial communities of a single spontaneous cocoa bean fermentation process sample.

    Science.gov (United States)

    Illeghems, Koen; Weckx, Stefan; De Vuyst, Luc

    2015-09-01

    A high-resolution functional metagenomic analysis of a representative single sample of a Brazilian spontaneous cocoa bean fermentation process was carried out to gain insight into its bacterial community functioning. By reconstruction of microbial meta-pathways based on metagenomic data, the current knowledge about the metabolic capabilities of bacterial members involved in the cocoa bean fermentation ecosystem was extended. Functional meta-pathway analysis revealed the distribution of the metabolic pathways between the bacterial members involved. The metabolic capabilities of the lactic acid bacteria present were most associated with the heterolactic fermentation and citrate assimilation pathways. The role of Enterobacteriaceae in the conversion of substrates was shown through the use of the mixed-acid fermentation and methylglyoxal detoxification pathways. Furthermore, several other potential functional roles for Enterobacteriaceae were indicated, such as pectinolysis and citrate assimilation. Concerning acetic acid bacteria, metabolic pathways were partially reconstructed, in particular those related to responses toward stress, explaining their metabolic activities during cocoa bean fermentation processes. Further, the in-depth metagenomic analysis unveiled functionalities involved in bacterial competitiveness, such as the occurrence of CRISPRs and potential bacteriocin production. Finally, comparative analysis of the metagenomic data with bacterial genomes of cocoa bean fermentation isolates revealed the applicability of the selected strains as functional starter cultures. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Detection of Cryptosporidium sp infection by PCR and modified acid fast staining from potassium dichromate preserved stool

    Directory of Open Access Journals (Sweden)

    Agnes Kurniawan

    2009-09-01

    Full Text Available Aim To identify the frequency of Cryptosporidium infection in children below 3 years old by examining concentrated long term preserved stool using PCR detection of 18S rRNA gene and compared with modified acid fast staining technique.Methods Hundred eighty eight stools from children ≤ 3 years old were stored for 13 months in 2.5% K2Cr2O7 solution at 40C. Cryptosporidium oocysts were isolated by water-ether concentration technique. The concentrates were smeared onto object glass and stained with modified acid fast staining, and the rest of the concentrates were DNA extracted by freezing and thawing cycles and proteinase K digestion, then direct PCR was done to detect 18S rRNA gene.Result The proportion of positive stools for Cryptosporidium sp by acid fast staining from concentrated stools and 18S rRNA PCR were 4.8% and 34.6% respectively, which showed statistically significant difference.Conclusion The frequency of Cryptosporidium infection among children ≤ 3 years old was very high and stool storage in K2Cr2O7 for 13 months did not affect the PCR result. High prevalence of Cryptosporidium infection indicated high transmission in that area and the potential to be transmitted to other individuals such as the immunocompromised. (Med J Indones 2009;18:147-52Key words: 18S rRNA, cryptosporidiosis

  13. Stool cultures for Shigella spp: improved specificity by using MacConkey agar with xylose.

    Science.gov (United States)

    Altwegg, M; Buser, J; von Graevenitz, A

    1996-03-01

    A total of 678 stool specimens were cultured on four different agars: on xylose-lysine-desoxycholate agar (XLD), MacConkey agar (Mac), MacConkey agar supplemented with xylose (Mac-X), and Hektoen enteric agar (HE). Isolation rates for shigellae were 77% on HE, 86% on Mac and Mac-X, and 91% on XLD. The specificities of the media were 61% for Mac, 75% for HE, and 78% for XLD and Mac-X. After overnight incubation, Mac-X is much easier to read than XLD, which requires incubation for at least 22 hours. Based on these results and also on the practical aspect that incubation for 22-21 hours does not fit well in our schedule, we now use Mac-X whenever shigellae need to be looked for (i.e. mainly patients with recent travel to tropical countries). As compared to our previous procedure the workload in the laboratory could be reduced by about 20%.

  14. Helicobacter pylori Seropositivity and Stool Antigen in Patients With Hyperemesis Gravidarum

    Science.gov (United States)

    Sinan Karadeniz, R.; Ozdegirmenci, Ozlem; Metin Altay, M.; Solaroglu, Ayse; Dilbaz, Serdar; Hızel, Nedret; Haberal, Ali

    2006-01-01

    The objective of this paper is to investigate whether Helicobacter pylori is an etiologic factor in hyperemesis gravidarum. Thirty one patients with hyperemesis gravidarum and twenty nine pregnant controls without hyperemesis gravidarum were included in this prospective study. All pregnant women were examined both for Helicobacter pylori serum immunoglobulin G antibodies (HpIgG Ab), showing chronic infection, and Helicobacter pylori stool antigens (HpSA), showing active gastrointestinal colonization. Chi-square and Student t tests were used accordingly for statistical analysis. Helicobacter pylori seropositivity was 67.7% in the patients with hyperemesis gravidarum and 79.3% in the control group (χ2 = 1.02, P = .31). HpSA was detected in 22.6% of patients with hyperemesis gravidarum, whereas 6.9% of patients in the control group. The difference was not statistically significant (χ2 = 2.89, P = .08). In this study, no relation was found between Helicobacter pylori and hyperemesis gravidarum. The low social status of women in both groups could be one of the reasons for the high prevalence of Hp infection. PMID:17093356

  15. Helicobacter pylori Seropositivity and Stool Antigen in Patients With Hyperemesis Gravidarum

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available The objective of this paper is to investigate whether Helicobacter pylori is an etiologic factor in hyperemesis gravidarum. Thirty one patients with hyperemesis gravidarum and twenty nine pregnant controls without hyperemesis gravidarum were included in this prospective study. All pregnant women were examined both for Helicobacter pylori serum immunoglobulin G antibodies (HpIgG Ab, showing chronic infection, and Helicobacter pylori stool antigens (HpSA, showing active gastrointestinal colonization. Chi-square and Student t tests were used accordingly for statistical analysis. Helicobacter pylori seropositivity was 67.7% in the patients with hyperemesis gravidarum and 79.3% in the control group ( χ 2 =1.02,P=.31 . HpSA was detected in 22.6% of patients with hyperemesis gravidarum, whereas 6.9% of patients in the control group. The difference was not statistically significant ( χ 2 =2.89,P=.08 . In this study, no relation was found between Helicobacter pylori and hyperemesis gravidarum. The low social status of women in both groups could be one of the reasons for the high prevalence of Hp infection.

  16. Stool-specimen testing practices adopted by clinical microbiology laboratories in the Veneto Region, Italy

    Directory of Open Access Journals (Sweden)

    Paolo Spolaore

    2008-03-01

    Full Text Available In order to correctly analyze data of laboratory diagnoses of infectious gastroenteritis for epidemiological purposes, a survey on analytical methods applied by hospital-based clinical microbiology laboratories has been conducted in the Veneto Region (Italy. The survey has been carried out in 2005 through a questionnaire collecting data on laboratory protocols and materials used for faecal specimens analysis. Laboratories from all the Local Health Units and University Hospitals of the Region returned the questionnaire. Almost all the laboratories routinely tested for the main foodborne pathogens: 23/23 for Salmonella, 22/23 for Shigella and 19/23 for Campylobacter jejuni. A great variety of analytical methods was applied for pathogen isolation; among these is worth of notice the inappropriate use of selenite broth for Shigella enrichment.Among noncultural methods, immunoassays were largely adopted. The survey allowed to appraise stool-specimen testing practices among laboratories of the Veneto Region; overall the compliance with guidelines proposed by the main national and international scientific societies resulted rather good.

  17. Endoscopy in acquired immunodeficiency syndrome patients with diarrhea and negative stool studies.

    Science.gov (United States)

    Wei, S C; Hung, C C; Chen, M Y; Wang, C Y; Chuang, C Y; Wong, J M

    2000-04-01

    Diarrhea is a frequent gastrointestinal symptom in patients with acquired immuno-deficiency syndrome (AIDS) and is a major source of morbidity and mortality. A stepwise diagnostic approach is often recommended to search for treatable causes. However, whether the stepwise diagnostic approach is adequate for planning treatment and whether specific treatment for infectious etiologies will affect the survival of patients with AIDS remain unknown. From March 1996 to September 1997, endoscopy was performed in AIDS patients with diarrhea, the etiology of which was not identified by noninvasive methods. Specific treatment was given according to the identified etiologies and symptomatic treatment was given for those without definite diagnosis. The clinical symptoms, signs, and duration of follow-up were recorded and survival patterns were analyzed. Etiologic diagnoses were made in 26 of 40 patients (65%) who underwent endoscopic studies. Amebic colitis and cytomegalovirus colitis were the 2 leading causes of prolonged diarrhea in patients with AIDS. Thirty-five patients (87.5%) recovered after treatment. The difference in survival time after diarrhea between patients whose symptoms resolved after treatment and those who continued to have diarrhea was statistically significant (p AIDS patients who had negative stool studies and did not respond to 2 weeks of empiric treatment. Specific treatment according to the results of endoscopy may improve survival in these patients.

  18. Combining single-particle inductively coupled plasma mass spectrometry and X-ray absorption spectroscopy to evaluate the release of colloidal arsenic from environmental samples.

    Science.gov (United States)

    Gomez-Gonzalez, Miguel Angel; Bolea, Eduardo; O'Day, Peggy A; Garcia-Guinea, Javier; Garrido, Fernando; Laborda, Francisco

    2016-07-01

    Detection and sizing of natural colloids involved in the release and transport of toxic metals and metalloids is essential to understand and model their environmental effects. Single-particle inductively coupled plasma mass spectrometry (SP-ICP-MS) was applied for the detection of arsenic-bearing particles released from mine wastes. Arsenic-bearing particles were detected in leachates from mine wastes, with a mass-per-particle detection limit of 0.64 ng of arsenic. Conversion of the mass-per-particle information provided by SP-ICP-MS into size information requires knowledge of the nature of the particles; therefore, synchrotron-based X-ray absorption spectroscopy (XAS) was used to identify scorodite (FeAsO4·2H2O) as the main species in the colloidal particles isolated by ultrafiltration. The size of the scorodite particles detected in the leachates was below 300-350 nm, in good agreement with the values obtained by TEM. The size of the particles detected by SP-ICP-MS was determined as the average edge of scorodite crystals, which show a rhombic dipyramidal form, achieving a size detection limit of 117 nm. The combined use of SP-ICP-MS and XAS allowed detection, identification, and size determination of scorodite particles released from mine wastes, suggesting their potential to transport arsenic. Graphical abstract Analytical approach for the detection and size characterization of As-bearing particles by SP-ICP-MS and XAS in environmental samples.

  19. Chicken single-chain antibody fused to alkaline phosphatase detects Aspergillus pathogens and their presence in natural samples by direct sandwich enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Xue, Sheng; Li, He-Ping; Zhang, Jing-Bo; Liu, Jin-Long; Hu, Zu-Quan; Gong, An-Dong; Huang, Tao; Liao, Yu-Cai

    2013-11-19

    A sensitive and specific analytical method to detect ubiquitous aflatoxigenic Aspergillus pathogens is essential for monitoring and controlling aflatoxins. Four highly reactive chicken single-chain variable fragments (scFvs) against soluble cell wall proteins (SCWPs) from Aspergillus flavus were isolated by phage display. The scFv antibody AfSA4 displayed the highest activity toward both A. flavus and A. parasiticus and specifically recognized a surface target of their cell walls as revealed by immunofluorescence localization. Molecular modeling revealed a unique compact motif on the antibody surface mainly involving L-CDR2 and H-CDR3. As measured by surface plasmon resonance, AfSA4 fused to alkaline phosphatase had a higher binding capability and 6-fold higher affinity compared with AfSA4 alone. Immunoblot analyses showed that the fusion had good binding capacity to SCWP components from the two fungal species. Direct sandwich enzyme-linked immunosorbent assays with mouse antiaspergillus monoclonal antibody mAb2A8 generated in parallel as a capture antibody revealed that the detection limit of the two fungi was as low as 10(-3) μg/mL, 1000-fold more sensitive than that reported previously (1 μg/mL). The fusion protein was able to detect fungal concentrations below 1 μg/g of maize and peanut grains in both artificially and naturally contaminated samples, with at least 10-fold more sensitivity than that reported (10 μg/g) thus far. Thus, the fusion can be applied in rapid, simple, and specific diagnosis of Aspergillus contamination in field and stored food/feed commodities.

  20. 3D-TROSY-based backbone and ILV-methyl resonance assignments of a 319-residue homodimer from a single protein sample

    Energy Technology Data Exchange (ETDEWEB)

    Krejcirikova, Anna; Tugarinov, Vitali, E-mail: vitali@umd.edu [University of Maryland, Department of Chemistry and Biochemistry (United States)

    2012-10-15

    The feasibility of practically complete backbone and ILV methyl chemical shift assignments from a single [U-{sup 2}H,{sup 15}N,{sup 13}C; Ile{delta}1-{l_brace}{sup 13}CH{sub 3}{r_brace}; Leu,Val-{l_brace}{sup 13}CH{sub 3}/{sup 12}CD{sub 3}{r_brace}]-labeled protein sample of the truncated form of ligand-free Bst-Tyrosyl tRNA Synthetase (Bst-{Delta}YRS), a 319-residue predominantly helical homodimer, is established. Protonation of ILV residues at methyl positions does not appreciably detract from the quality of TROSY triple resonance data. The assignments are performed at 40 Degree-Sign C to improve the sensitivity of the measurements and alleviate the overlap of {sup 1}H-{sup 15}N correlations in the abundant {alpha}-helical segments of the protein. A number of auxiliary approaches are used to assist in the assignment process: (1) selection of {sup 1}H-{sup 15}N amide correlations of certain residue types (Ala, Thr/Ser) that simplifies 2D {sup 1}H-{sup 15}N TROSY spectra, (2) straightforward identification of ILV residue types from the methyl-detected 'out-and-back' HMCM(CG)CBCA experiment, and (3) strong sequential HN-HN NOE connectivities in the helical regions. The two subunits of Bst-YRS were predicted earlier to exist in two different conformations in the absence of ligands. In agreement with our earlier findings (Godoy-Ruiz in J Am Chem Soc 133:19578-195781, 2011), no evidence of dimer asymmetry has been observed in either amide- or methyl-detected experiments.

  1. Robustness of two single-item self-esteem measures: cross-validation with a measure of stigma in a sample of psychiatric patients.

    Science.gov (United States)

    Bagley, Christopher

    2005-08-01

    Robins' Single-item Self-esteem Inventory was compared with a single item from the Coopersmith Self-esteem. Although a new scoring format was used, there was good evidence of cross-validation in 83 current and former psychiatric patients who completed Harvey's adapted measure of stigma felt and experienced by users of mental health services. Scores on the two single-item self-esteem measures correlated .76 (p self-esteem in users of mental health services.

  2. Performance enhancement of the single-phase series active filter by employing the load voltage waveform reconstruction and line current sampling delay reduction methods

    DEFF Research Database (Denmark)

    Senturk, O.S.; Hava, A.M.

    2011-01-01

    This paper proposes the waveform reconstruction method (WRM), which is utilized in the single-phase series active filter's (SAF's) control algorithm, in order to extract the load harmonic voltage component of voltage harmonic type single-phase diode rectifier loads. Employing WRM and the line...

  3. Early Adoption of a Multitarget Stool DNA Test for Colorectal Cancer Screening.

    Science.gov (United States)

    Finney Rutten, Lila J; Jacobson, Robert M; Wilson, Patrick M; Jacobson, Debra J; Fan, Chun; Kisiel, John B; Sweetser, Seth; Tulledge-Scheitel, Sidna M; St Sauver, Jennifer L

    2017-05-01

    To characterize early adoption of a novel multitarget stool DNA (MT-sDNA) screening test for colorectal cancer (CRC) screening and to test the hypothesis that adoption differs by demographic characteristics and prior CRC screening behavior and proceeds predictably over time. We used the Rochester Epidemiology Project research infrastructure to assess the use of the MT-sDNA screening test in adults aged 50 to 75 years living in Olmsted County, Minnesota, in 2014 and identified 27,147 individuals eligible or due for screening colonoscopy from November 1, 2014, through November 30, 2015. We used electronic Current Procedure Terminology and Health Care Common Procedure codes to evaluate early adoption of the MT-sDNA screening test in this population and to test whether early adoption varies by age, sex, race, and prior CRC screening behavior. Overall, 2193 (8.1%) and 974 (3.6%) individuals were screened by colonoscopy and MT-sDNA, respectively. Age, sex, race, and prior CRC screening behavior were significantly and independently associated with MT-sDNA screening use compared with colonoscopy use after adjustment for all other variables (Padoption of MT-sDNA screening increased over time and were highest in those aged 50 to 54 years, women, whites, and those who had a history of screening. The use of the MT-sDNA screening test varied predictably by insurance coverage. The rates of colonoscopy decreased over time, whereas overall CRC screening rates remained steady. The results of the present study are generally consistent with predictions derived from prior research and the diffusion of innovation framework, pointing to increasing use of the new screening test over time and early adoption by younger patients, women, whites, and those with prior CRC screening. Copyright © 2017 Mayo Foundation for Medical Education and Research. Published by Elsevier Inc. All rights reserved.

  4. Occurrence of virulent and antibiotic-resistant Shiga toxin-producing Escherichia coli in some food products and human stool in Egypt.

    Science.gov (United States)

    Hamed, Osman Mohamed; Sabry, Maha Ahmed; Hassanain, Nawal A; Hamza, Eman; Hegazi, Ahmed G; Salman, Marwa Badawy

    2017-10-01

    Shiga toxin-producing Escherichia coli (STEC) represent a severe public health issue worldwide, causing life-threatening diseases in the human gastrointestinal tract. This study aimed to determine the occurrence of virulent and antibiotic-resistant STEC in retail meat and milk products and human stool samples and to characterize the genes encoding for virulence and antibiotic resistance among the identified STEC isolates. A total of 260 food samples were randomly collected from retail markets in different localities of El Giza Governorate, Egypt. 50 stool specimens were obtained from children that had diarrhea at Embaba Fever Hospital. All collected samples were initially subjected to bacteriological examination and serotyping, and then subsequently, the isolates were exposed to polymerase chain reaction application and sequencing for the identification of the virulence-related genes. Finally, the virulent STEC isolates were tested for antibiotic susceptibility. Serotyping of the 76 biochemically identified isolates showed that 18 were STEC with a predominance of non-O157 (16) while 2 O157:K-serotype was detected only in one food and one human isolate. Molecular identification of the virulence genes illustrated that the minced meat showed the highest prevalence of STEC (8%) as compared to the other food products. In the humans, the O157 was the only serotype that expresses the Shiga toxin-associated gene ( eaeA ). Antibiotic susceptibility test displayed that 13 of the 17 food and human isolates (76.47%) were resistant to cephalothin (KF30). 9 of the 13 cephalothin-resistant isolates harbor the β lactamase ( bla TEM )-resistant gene. All isolates were sensitive to chloramphenicol, ciprofloxacin, amikacin, and gentamicin. DNA sequencing and phylogenetic analysis of the stx2- positive minced meat isolate revealed a high genetic relatedness with beef minced meat from the USA and Australia. This study showed the predominance of non-O157 among the identified isolates

  5. Structural and EPR studies on single-crystal and polycrystalline samples of copper(II) and cobalt(II) complexes with N2S2-based macrocyclic ligands.

    Science.gov (United States)

    Tamayo, Abel; Casabó, Jaume; Escriche, Lluís; González, Pablo; Lodeiro, Carlos; Rizzi, Alberto C; Brondino, Carlos D; Passeggi, M C G; Kivekäs, Raikko; Sillanpää, Reijo

    2007-07-09

    The properties of Cu(II) and Co(II) complexes with oxygen- or nitrogen-containing macrocycles have been extensively studied; however, less attention has been paid to the study of complexes containing sulfur atoms in the first coordination sphere. Herein we present the interaction between these two metal ions and two macrocyclic ligands with N2S2 donor sets. Cu(II) and Co(II) complexes with the pyridine-containing 14-membered macrocycles 3,11-dithia-7,17-diazabicyclo[11.3.1]heptadeca-1(17),13,15-triene (L) and 7-(9-anthracenylmethyl)-3,11-dithia-7,17-diazabicyclo[11.3.1]heptadeca-1(17),13,15-triene (L1) have been synthesized. The X-ray structural analysis of {[Co(ClO4)(H2O)(L)][Co(H2O)2(L)]}(ClO4)3 shows two different metal sites in octahedral coordination. The EPR spectra of powdered samples of this compound are typical of distorted six-coordinated Co(II) ions in a high-spin (S=3/2) configuration, with the ground state being S=1/2 (g1=5.20, g2=3.20, g3=1.95). The EPR spectrum of [Cu(ClO4)(L)](ClO4) was simulated assuming an axial g tensor (g1=g2=2.043, g3=2.145), while that of [Cu(ClO4)(L1)](ClO4) slightly differs from an axial symmetry (g1=2.025, g2=2.060, g3=2.155). These results are compatible with a Cu(II) ion in square-pyramidal coordination with N2S2 as basal ligands. Single-crystal EPR experiment performed on [Cu(ClO4)(L1)](ClO4) allowed determining the eigenvalues of the molecular g tensor associated with the copper site, as well as the two possible orientations for the tensor. On the basis of symmetry arguments, an assignment in which the eigenvectors are nearly along the Cu(II)-ligand bonds is chosen.

  6. Single-Molecule Counting of High-Sensitivity Troponin I in Patients Referred for Diagnostic Angiography: Results From the CASABLANCA (Catheter Sampled Blood Archive in Cardiovascular Diseases) Study.

    Science.gov (United States)

    McCarthy, Cian P; Ibrahim, Nasrien E; Lyass, Asya; Li, Yiwei; Gaggin, Hanna K; Simon, Mandy L; Mukai, Renata; Gandhi, Parul; Kelly, Noreen; Motiwala, Shweta R; van Kimmenade, Roland R J; Massaro, Joseph M; D'Agostino, Ralph B; Januzzi, James L

    2018-03-08

    The meaning of high-sensitivity troponin I (hsTnI) concentrations in patients without acute myocardial infarction (MI) requires clarity. We hypothesized that among patients referred for diagnostic coronary angiography without acute MI, hsTnI concentrations would correlate with prevalent coronary artery disease (CAD) and predict incident cardiovascular events and mortality. We measured hsTnI using a single-molecule counting assay (99th percentile, 6 ng/L) in samples from 991 patients obtained at the time of angiography. Concentrations of hsTnI were assessed relative to the severity of CAD and prognosis during mean follow-up of 3.7 years. Median hsTnI concentration was 4.19 ng/L; 38% of patients had hsTnI concentrations ≥99th percentile. Across increasing hsTnI quartiles, patients had higher prevalence of angiographic CAD; in multivariate models, hsTnI ≥99th percentile independently predicted obstructive CAD (odds ratio: 2.57; P 70% coronary stenosis, hsTnI ≥99th percentile independently predicted incident MI (HR: 1.87; P =0.01), cardiovascular mortality (HR: 2.74; P =0.001), and the composite end point of MI and all-cause death (HR: 2.06; P <0.001). In participants with coronary stenosis <70%, hsTnI ≥99th percentile even more strongly predicted incident MI (HR: 8.41; P <0.001), cardiovascular mortality (HR: 3.60; P =0.03), and the composite end point of MI and all-cause death (HR: 3.62; P <0.001). In a large prospective cohort of patients who were free of prevalent MI and undergoing diagnostic coronary angiography, hsTnI concentrations were associated with higher prevalence of CAD and predicted incident MI, cardiovascular death, and all-cause death. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00842868. © 2018 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

  7. Identification of Shigella species in stool specimens by DNA amplification of different loci of the Shigella virulence plasmid.

    Science.gov (United States)

    Yavzori, M; Cohen, D; Wasserlauf, R; Ambar, R; Rechavi, G; Ashkenazi, S

    1994-03-01

    The sensitivity and specificity of the polymerase chain reaction (PCR) for detection of DNA sequences specific to Shigella spp. and enteroinvasive Escherichia coli (EIEC) in stools was evaluated. Stool specimens were obtained from patients with acute gastroenteritis before and after antibiotic treatment. Fecal material was pre-incubated in phosphate-buffered saline, gram-negative broth or brain heart infusion (BHI) broth, and DNA was extracted and amplified. Primers complementary to the ial or the virF loci of the 140 MDa plasmid of Shigella were evaluated. The highest sensitivity for detection of Shigella DNA in stools (higher than that of culture) was reached by pre-incubation of the fecal material in BHI broth and use of virF primers for amplification. The specificity of this PCR protocol was documented by the negative results obtained with non-Shigella enteric organisms. These findings point out the important diagnostic and epidemiologic potential of the virF-specific PCR protocol in the investigation of Shigella infections.

  8. Treatment of severe cholera: a review of strategies to reduce stool output and volumes of rehydration fluid.

    Science.gov (United States)

    Butler, Thomas

    2017-05-01

    Severe cholera is a life-threatening illness of hypovolemic shock and metabolic acidosis due to rapid and profuse diarrheal fluid loss. Emergency life-saving therapy is i.v. saline, optionally supplemented with potassium and alkali to correct the fluid deficit, potassium losses and acidosis. After this initial rehydration, for the next 2 days ongoing stool losses are replaced with oral rehydration solution (ORS), which contains sodium chloride, potassium and alkali together with glucose or rice powder as a source of glucose to serve as a carrier for sodium. In actual field trials, antibiotics are given to reduce fluid requirements, but large volumes averaging about 7 liters of i.v. fluid followed by about 14 liters of ORS have been given to adult patients. Disturbing trends during therapy have included overhydration, hyponatremia and polyuria. It is suggested that stool output and fluid requirements could be reduced, if borne out in future research, by avoiding overhydration by restricting ORS intake to match stool output and promoting intestinal reabsorption of luminal fluid by early introduction of glucose without salts into the intestine, more gradual correction of dehydration, giving mineralocorticoid and vasopressin, and infusing glucose or short-chain fatty acids into the proximal colon. © The Author 2017. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Pre-operative stool analysis for intestinal parasites and fecal occult blood in patients with acute appendicitis.

    Science.gov (United States)

    Hatipoğlu, Sinan; Lök, Uğur; Gülaçtı, Umut; Çelik, Tuncay

    2016-09-01

    Etiology of acute appendicitis (AA) rarely involves parasitic infections of gastrointestinal (GI) tract. Preoperative diagnosis of parasitic infections in appendix remains difficult, although parasites can sometimes be observed inside the lumen during histopathological examination. The aim of the present study was to prospectively screen prevalence and species of intestinal parasites and adherence of fecal occult blood (FOB) in patients admitted to emergency department (ED) with clinical symptoms of AA who underwent appendectomy. Demographic and stool analysis data of a total of 136 patients (≥13 years old) who underwent appendectomy between July 2009 and December 2014 were prospectively assessed, and histopathological data of all patients were retrospectively assessed. In histopathological examination after appendectomy, of 136 patients, 75.5% (n=103) had AA, 11.1% (n=15) had perforated appendicitis (PA), and 13.2% (n=18) had a negative appendicitis (normal appendix, NA). Pre-operative stool analysis revealed that 25% (n=34) had intestinal parasites and 14.7% (n=20) of patients had positive fecal occult blood test (FOBT). Those with positive FOBT represented 9.7% (n=10) of 103 AA patients, 53.3% (n=8) of 15 PA patients, and 11.1% (n=2) of 18 NA patients; this was statistically more significant for PA than other groups (pparasites in stool might not be associated with appendicitis, but it can occasionally lead to pathological findings of appendicitis. A positive FOBT may be a predictor for PA.

  10. Evaluation of the TRCRtest NV-W for norovirus detection in stools by the Transcription-Reverse Transcription Concerted method.

    Science.gov (United States)

    Medici, Maria Cristina; Tummolo, Fabio; Albonetti, Valeria; Pinardi, Federica; Ferraglia, Francesca; Chezzi, Carlo; Arcangeletti, Maria Cristina; De Conto, Flora; Calderaro, Adriana

    2013-11-01

    A novel molecular assay, TRCRtest NV-W, based on a transcription-reverse transcription concerted reaction (TRC) for isothermal amplification and real-time detection of norovirus in stools was assessed and compared with an RT-nPCR. Archived stools positive for either different types or variants of norovirus genogroups I and II or other enteric viruses were used to assess the sensitivity and specificity of the novel assay. The TRC assay was 100% specific since it detected all the noroviruses tested and it did not display cross reactivity with other enteric viruses. When screening a collection of 387 stools with the TRC and RT-nPCR assays, the TRC displayed concordance, sensitivity, specificity, positive and negative predictive values of 96.6%, 81%, 99.7%, 98.1%, and 96.3%, respectively, after retesting the negative specimens. Additional PCRs and/or sequencing, used to understand inconsistent results between TRC and RT-nPCR, confirmed all positive results and did not reveal nucleotide variations in the TRC probe and primers binding sites. The TRC assay may be a rapid and ease of use tool for the detection of noroviruses in clinical virology laboratories even in the face of rapidly evolving noroviruses. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Prevalence of Shiga toxin-producing and enteropathogenic Escherichia coli marker genes in diarrhoeic stools in a New Zealand catchment area.

    Science.gov (United States)

    Thomas, Rowan R; Brooks, Heather J L; O'Brien, Rory

    2017-01-01

    Shiga toxin-producing (STEC) and enteropathogenic (EPEC) Escherichia coli are gastrointestinal pathogens causing diarrhoeal and extraintestinal disease. Due to lack of EPEC screening and use of Sorbitol-MacConkey (SMAC) agar in faecal screening, the true prevalence of EPEC and non-O157 STEC in New Zealand diarrhoeal cases is unknown. Diarrhoeic stools sourced from Dunedin hospital were pre-enriched, DNA extracted with Chelex-100 resin and screened using a multiplex TaqMan quantitative PCR assay amplifying stx1, sxt2 and EPEC (eae) gene markers. Of the 522 diarrhoeic samples surveyed, 8 (1.53%) were PCR positive for stx1/stx2 and 23 (4.41%) were positive for eae. Six (75%) of the stx + samples were uncommon non-O157 serotypes, and the remainder were found to be positive for both O103 and O157 STEC somatic antigens. Results revealed shortcomings in current screening protocols for pathogenic E. coli; SMAC is not sufficiently discriminatory to detect emergent STEC serotypes and EPEC likely has an unappreciated role in cases of diarrhoea in New Zealand. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  12. Interaction of z component of magnetic field between two samples of GO material in the round rotational single sheet tester (RRSST)

    International Nuclear Information System (INIS)

    Gorican, Viktor; Hamler, Anton; Jesenik, Marko; Stumberger, Bojan; Trlep, Mladen

    2006-01-01

    The magnetic properties of two grain-oriented (GO) samples of the same grade were measured under alternating and rotational magnetic flux conditions. Two samples were measured separately and then together in different arrangement to each other. The interaction of magnetic field between two samples were measured by using a coil, which was placed in between. The results show that the H z component influence measured magnetic properties in the x-y plane

  13. and improvement of bowel function by increasing stool frequency pursuant to Article 13(5) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    2014-01-01

    to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improvement of bowel function by increasing stool frequency. The food that is the subject of the health claim is a combination of four bacterial strains—B. longum...... proposed by the applicant is "improves stool frequency". The Panel considers that improvement of bowel function by increasing stool frequency, provided that it does not result in diarrhoea, is a beneficial physiological effect. The Panel considers that the human study provided for the substantiation...... of the claim did not find an increase in stool frequency following consumption of a combination of the bacterial strains which is the subject of the claim. The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of B. longum LA 101, L...

  14. Intestinal Parasite Profile in the Stool of HIV Positive Patients in relation to Immune Status and Comparison of Various Diagnostic Techniques with Special Reference to Cryptosporidium at a Tertiary Care Hospital in South India

    Directory of Open Access Journals (Sweden)

    Vishnu Kaniyarakkal

    2016-01-01

    Full Text Available Acquired immunodeficiency syndrome and related opportunistic infections are a significant cause of morbidity and mortality in susceptible population. This study aims to negate the paucity of data regarding the relation between CD4 levels, prevalence of enteric parasites, and the outcome of treatment with HAART (highly active antiretroviral therapy and Cotrimoxazole in Kerala, India. Multiple stool samples from 200 patients in a cross-sectional study were subjected to microscopy and Cryptosporidium stool antigen ELISA. Parasites were identified in 18 samples (9%. Cystoisospora and Cryptosporidium spp. were seen in 9 cases (4.5% and 5 cases (2.5%, respectively. Microsporidium spores and Chilomastix mesnili cysts were identified in 1 case each (0.5% each. Seven cases of Cystoisospora diarrhoea recovered after treatment with Cotrimoxazole. Diarrhoea due to Cryptosporidium spp. in all 5 cases subsided after immune reconstitution with HAART. This study concludes that a positive association was seen between low CD4 count (<200 cells/μL and overall parasite positivity (P value < 0.01. ELISA is a more sensitive modality for the diagnosis of Cryptosporidium diarrhoea. Chilomastix mesnili, generally considered a nonpathogen, may be a cause of diarrhoeal disease in AIDS. Immune reconstitution and Cotrimoxazole prophylaxis remain to be the best therapeutic approach in AIDS-related diarrhoea.

  15. Intestinal Parasite Profile in the Stool of HIV Positive Patients in relation to Immune Status and Comparison of Various Diagnostic Techniques with Special Reference to Cryptosporidium at a Tertiary Care Hospital in South India.

    Science.gov (United States)

    Kaniyarakkal, Vishnu; Mundangalam, Nizamuddin; Moorkoth, Anitha Puduvail; Mathew, Sheela

    2016-01-01

    Acquired immunodeficiency syndrome and related opportunistic infections are a significant cause of morbidity and mortality in susceptible population. This study aims to negate the paucity of data regarding the relation between CD4 levels, prevalence of enteric parasites, and the outcome of treatment with HAART (highly active antiretroviral therapy) and Cotrimoxazole in Kerala, India. Multiple stool samples from 200 patients in a cross-sectional study were subjected to microscopy and Cryptosporidium stool antigen ELISA. Parasites were identified in 18 samples (9%). Cystoisospo