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Artykuły w czasopismach na temat "Blood samples and stains"
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Buchanan, Ruaridh, Stanley Fan i Caoimhe NicFhogartaigh. "Performance of Gram Stains and 3 Culture Methods in the Analysis of Peritoneal Dialysis Fluid". Peritoneal Dialysis International: Journal of the International Society for Peritoneal Dialysis 39, nr 2 (marzec 2019): 190–92. http://dx.doi.org/10.3747/pdi.2018.00087.
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Microbiological diagnosis of peritoneal dialysis (PD)-related peritonitis includes PD fluid cell count, Gram stain, and culture, as recommended by the International Society for Peritoneal Dialysis. In this retrospective study, we examined the utility of Gram stains and compared 3 culture methods. We examined a laboratory cohort (samples sent to the laboratory for any reason; n = 251) and a clinical cohort (samples sent from patients felt clinically to have peritonitis; n = 264). Culture positivity rates were higher in the clinical cohort (39.4%) than the laboratory cohort (21.5%), with no difference in the distribution of organisms between the cohorts; cell counts were significantly higher in culture-positive samples in both cohorts. Rates of positivity in the laboratory and clinical cohorts, respectively, were as follows: Gram stains 1.9% and 7.7%; direct plate culture 13% and 30.8% and “bedside” inoculated blood culture bottles 82.1% and 92.8%. Enrichment culture was never negative when another method was positive. Our data indicate that enrichment culture can be used as a single culture methodology for analyzing PD fluid without loss of sensitivity. They also suggest that Gram stains are of relatively low yield; consideration could be given to ceasing their routine performance provided that broad antimicrobial therapy is administered pending culture results.
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Yudianto, Ahmad, i Yeti Eka Sispitasari. "DNA ISOLATION FROM HUMAN URINE STAIN AS AN ALTERNATIVE MATERIAL FOR PERSONAL IDENTIFICATION EXAMINATION". Folia Medica Indonesiana 52, nr 4 (14.08.2017): 277. http://dx.doi.org/10.20473/fmi.v52i4.5476.
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Accurate determination of personal identity is crucial for an investigation since any inaccuracy may lead to fatal consequences in the judicial process. Identification through DNA analysis involves somatic chromosomes and mtDNA. Each part of the human body can be taken as a specimen since every nucleated cell in the body of an individual has identical DNA sequence. To date, samples for identification through DNA analysis are obtained from blood stains, semen stains, bones, vaginal swab, buccal swab etc. In certain cases, urine stains on the clothing have frequently been overlooked. So far, personal identification through DNA analysis by the use of urine stains has not been commonly carried out. The present study detected bands in the loci CSF1PO, THO1, TPOX and 106bp-112bp amelogenin in all samples visualized from the results of Polymerase Chain Reaction (PCR) with Polyacrylamid Agarose Gel Electrophoreses-silver staining for exposure durations of 1, 7 and 14 days. However, for exposure duration of 20 days (the maximum in the study), bands were only detected in the loci THO1 and TPOX in all samples (100%), whereas the loci CSF1PO and 50% amelogenin exhibited obvious bands. This indicated that DNA analysis of urine stains through detection of the locus STR CSF1PO, THO1, TPOX exhibited different detection responses for different exposure durations assigned to the samples of urine stain. Successful detection of these loci was supported by the differences in amplicon product and GC content at each locus. Of the loci studied, the ratio of GC content of the primers, sorted from the lowest, were as follows: locus CSF1PO of 42.6 1%, TPOX of 56.25%, and THO1 of 63.83%. In conclusion, the loci THO1 and TPOX had the same probability of success in the STR examination compared with the locus CSF1PO.
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Lacerenza, Daniela, Giorgio Caudullo, Elena Chierto, Serena Aneli, Giancarlo Di Vella, Marco Barberis, Samuele Voyron, Paola Berchialla i Carlo Robino. "Evaluation of the Effects of Different Sample Collection Strategies on DNA/RNA Co-Analysis of Forensic Stains". Genes 13, nr 6 (30.05.2022): 983. http://dx.doi.org/10.3390/genes13060983.
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The aim of this study was to evaluate the impact of different moistening agents (RNase-free water, absolute anhydrous ethanol, RNAlater®) applied to collection swabs on DNA/RNA retrieval and integrity for capillary electrophoresis applications (STR typing, cell type identification by mRNA profiling). Analyses were conducted on whole blood, luminol-treated diluted blood, saliva, semen, and mock skin stains. The effects of swab storage temperature and the time interval between sample collection and DNA/RNA extraction were also investigated. Water provided significantly higher DNA yields than ethanol in whole blood and semen samples, while ethanol and RNAlater® significantly outperformed water in skin samples, with full STR profiles obtained from over 98% of the skin samples collected with either ethanol or RNAlater®, compared to 71% of those collected with water. A significant difference in mRNA profiling success rates was observed in whole blood samples between swabs treated with either ethanol or RNAlater® (100%) and water (37.5%). Longer swab storage times before processing significantly affected mRNA profiling in saliva stains, with the success rate decreasing from 91.7% after 1 day of storage to 25% after 7 days. These results may contribute to the future development of optimal procedures for the collection of different types of biological traces.
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Sun, Mao-ling, Ji-long Zheng, Bao-jie Wang i Jun Yao. "Sperm Cell Capture Based on ABH Antigen Differences to Separate Two Men in Mixed Seminal Stains". BioMed Research International 2021 (27.11.2021): 1–5. http://dx.doi.org/10.1155/2021/7269237.
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Personal identification of two individuals in mixed semen samples in forensic DNA testing in general usually involves analysis using autosomal and Y chromosome short tandem repeats (STRs). Results may exclude unrelated donors but cannot identify individuals. In this study, sperm cell capture based on ABH antigen differences was used to obtain the cells with the single ABO blood type. Immunohistochemical staining using labeled anti-A, anti-B, and anti-H antibodies and the laser microdissection system can be used to enrich sperm with different ABO types in mixed seminal stains from two individuals. Then, PCR amplification and capillary electrophoresis were performed to genotype the STR loci. To some extent, after sperm cell capture based on ABH antigen differences, autosomal STR typing using enriched single blood group cells can be utilized to partially identify different individuals in a mixed seminal stain sample from two individuals.
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Alama, Khalid, i Prof/ Adam Dawoud Abakar. "THE ACCURACY OF LIQUID BASE PAP SMEAR VS CONVENTIONAL PAP SMEAR CYTOLOGY". European Journal of Health Sciences 4, nr 1 (7.10.2019): 32. http://dx.doi.org/10.47672/ejhs.414.
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The main aim of the present study was to assess the diagnostic accuracy of Liquid base versus Conventional smears (CS). The specific objective was to evaluate and compare efficacy liquid base cytology with conventional cytology (CS) as a screening tool and to assess the quality of immunohistochemical stain in conventional smears. A prospective study including 100 cervical samples over a period of six month. Split sample was obtained using cervex-brush. CS was prepared from the brush and the brush head was suspended in the LBC vial and processed by thin prep 5000 machine. The smears were stained with Pap stain and extra five conventional and thin prep slides prepared and stained with immunomarker. Results showed that there were 4.0% unsatisfactory (U/S) cases in CS and 1.0% in LBC; the main cause was ranging between obscuring blood and inflammation in CS and low squamous cellularity in LBC. About 5% split samples had epithelial abnormalities both in CS and LBC (3% atypical squamous cells of undetermined significance (ASCUS), devided between LBS 2% while CS1%.Low grade squamous intraepithelial lesion (LSIL) 2%, devided between LBC 1% and CS 1%. Infections as Trichomonas vaginalis (TV) and spores of candida species, 1% and 2% respectively detected only in LBC smear and missed in CS preparations of the same samples, considering 2-3 minutes for LBC screening and 5-6 minutes for CS screening following the international standards. Conventional smears did not appear to confer a cytomorphological advantage and has a lower diagnostic accuracy using IHC. The sensitivity of Thin Prep was significantly higher than that of CS due to cellular clumps and presence of marked inflammatory cells and blood which compete other epithelial cellular elements in staining affinity in addition to the length of the smear which need large volume of stains to cover the whole area. While the confined area of thin prep smear and homogenous cellular distribution support the advantages of thin prep over the conventional smear when using IHC stain. The study concluded that LBC technique leads to significant reduction of U/S rate. LBC samples offered better clarity, uniform spread of smears, less time for screening and better handling of hemorrhagic and inflammatory samples. In addition to feasibility to do further special stains and HPV tests. LBC had equivalent sensitivity and specificity to CS.
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Maryani, Maryani, Desvita Sari, Ridha Wahyutomo i Masfiyah Masfiyah. "Errors in Interpretation of Gram Stain in The First Notification from Positive BACTEC Blood Cultures in Clinical Microbiology Laboratory of Dr. Kariadi Hospital". Sains Medika : Jurnal Kedokteran dan Kesehatan 4, nr 1 (8.12.2012): 23. http://dx.doi.org/10.30659/sainsmed.v4i1.381.
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Background: Blood cultures in conjunction with the initial Gram stain of positive cultures have often been considered the “gold standard†for the diagnosis of bacteremia. When blood cultures turn positive, the attending physicians are usually notified immediately about Gram stain findings. However, information on the accuracy of Gram staining is very limited. We examined the error of preliminary blood culture reports provided by a local laboratory in an observational study.Design and Method: This was an observational study with a cross sectional approach. In this study, 369 blood cultures were examined. The positive blood cultures (135 samples) were then examined by Gram stain. Blood cultures handled on Bactec 9050, while the Gram stain was done in standard procedure Gram. Interpretation errors of Gram stain were confirmed by cultures result.Result: During one month (April 2011) we examined 369 blood cultures which 135 are positive (36.5%). Positive blood cultures were misread for 6 (4.4%) of 135 patients, they were two read as gram positive cocci had gram negative organisms by culture which were Acinetobacter baumannii, one read as gram positive bacilli had gram negative bacilli by culture which was Klebsiella pneumoniae. One isolate read as gram negative bacilli had gram positive bacilli which was Bacillus species, while two sample read as gram negative bacilli only had polymicrobial by culture, of these one isolate grew to be Enterobacter aerogenes and Staphylococcus aureus and the other were Escherichia coli and Acinetobacter spp.Conclusion: The overall 4.4% error rate of misinterpreted Gram stains from positive blood culture bottles is relatively high, so laboratory professionals and clinical microbiologist must be aware of the potential types of error that occur (Sains Medika, 4(1):23-29).
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Mosiiuk, E., i O. Karasova. "Features of establishing the presence of blood in old stains by the method of thin-layer vertical chromatography". Theory and Practice of Forensic Science and Criminalistics 23, nr 1 (27.07.2021): 400–409. http://dx.doi.org/10.32353/khrife.1.2021.31.
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Due to the fact that the bloodstain pattern analysis takes the first place in the structure of the study of biological objects, the question of the study of blood patterns is relevant today. The main task (that will allow to solve the following, which puts the investigation before the experts), is to prove that these patterns, which are examined, contain blood, (regardless of their remoteness, attempts to destroy patterns of blood), find out its types or refute traces of blood. If the expert fails to prove the presence of blood, he must refuse to address the following issues. Otherwise, the expert may obtain results that will ultimately lead him to an erroneous conclusion.
The article considers options for extraction of old blood stains in order to establish the presence of blood by the method of thin-layer vertical chromatography. This question is still relevant, as biological material can change under the influence of environmental factors and time and this can be an obstacle to establishing the fact of its presence on physical evidence, and also it can do harm to the quality of the study and the correctness of the conclusions, made as a result of the study. In order to select substances, that could provide rapid extraction of old blood stains, a study was performed using weak solutions of alkalis and acids and subsequent determination of the presence of blood by thin layer vertical chromatography. In immunological studies, there have been cases where blood crusts on non-hygroscopic surfaces under the influence of natural factors (such as high temperatures and direct sun rays) have not been extracted in saline and distilled water for several days. In addition, when using known blood samples in the reactions, the dependence of the saturation of the extracts on the age of the samples and the time of their extraction. These observations prompted researches in the sector of biological research and accounting of the Cherkasy NDEKTs of the Ministry of Internal Affairs on methods for extracting old blood stains, which would allow to detect blood in the shortest possible time by the method of thin-layer vertical chromatography.
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Oliveira, Naila Francis Paulo de, Mary Anne Heidi Dolder i Selma Candelária Genari. "Light microscope observation of circulating human lymphocytes cultured in vitro". Brazilian Archives of Biology and Technology 53, nr 5 (październik 2010): 1097–100. http://dx.doi.org/10.1590/s1516-89132010000500013.
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The purpose of this work was to study the isolation and a light microscopy technique for cultured lymphocytes. Blood samples were obtained by venipuncture with an anticoagulant added and centrifuged in a Percoll density gradient to separate the leukocytes. Lymphocytes were placed in 25 cm ³ tissue culture flasks at 37ºC. After culturing, they were fixed and stained with the methods used for blood smears. Results showed that not all fixing solutions and stains were an equally good choice for cultured lymphocytes.
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Stazio, Alice, Juan G. Victores, David Estevez i Carlos Balaguer. "A Study on Machine Vision Techniques for the Inspection of Health Personnels’ Protective Suits for the Treatment of Patients in Extreme Isolation". Electronics 8, nr 7 (30.06.2019): 743. http://dx.doi.org/10.3390/electronics8070743.
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The examination of Personal Protective Equipment (PPE) to assure the complete integrity of health personnel in contact with infected patients is one of the most necessary tasks when treating patients affected by infectious diseases, such as Ebola. This work focuses on the study of machine vision techniques for the detection of possible defects on the PPE that could arise after contact with the aforementioned pathological patients. A preliminary study on the use of image classification algorithms to identify blood stains on PPE subsequent to the treatment of the infected patient is presented. To produce training data for these algorithms, a synthetic dataset was generated from a simulated model of a PPE suit with blood stains. Furthermore, the study proceeded with the utilization of images of the PPE with a physical emulation of blood stains, taken by a real prototype. The dataset reveals a great imbalance between positive and negative samples; therefore, all the selected classification algorithms are able to manage this kind of data. Classifiers range from Logistic Regression and Support Vector Machines, to bagging and boosting techniques such as Random Forest, Adaptive Boosting, Gradient Boosting and eXtreme Gradient Boosting. All these algorithms were evaluated on accuracy, precision, recall and F 1 score; and additionally, execution times were considered. The obtained results report promising outcomes of all the classifiers, and, in particular Logistic Regression resulted to be the most suitable classification algorithm in terms of F 1 score and execution time, considering both datasets.
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Shabbir, Sheeba, Nusrat Ali, Iftikhar ,. Ahmad, Aina Khurshid, Muhammad Farhan Siddiq Rao, Sabika Hussain i Muhammad Rizwan. "Enzyme Immunoassay for the Detection of Human Chorionic Gonadotropin (HCG) and Placental MRNA Marker a Practical Technique for Forensic Pregnancy Identification in Bloodstains". Pakistan Journal of Medical and Health Sciences 16, nr 10 (30.10.2022): 804–6. http://dx.doi.org/10.53350/pjmhs221610804.
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Introduction: The diagnosis of pregnancy from forensic bloodstains can be useful in cases of infanticide, criminal abortions and missing person identification. Objective: This research illustrated the use of a rapid, precise, and tremendously responsive enzyme immunoassay kit designed for medical usage, which is put to good use in our lab for qualitative HCG detection in blood stains and has proven to be a useful tool in forensic pregnancy identification. Methods: HCG concentrations had previously been generally known, and total eighty whole blood samples were taken: forty expectant females (every single one between months one to six of pregnancy), twenty healthy young males, and twenty postmenopausal females with good health ration has been cleared in all data. Results: Enzyme immunoassay is a useful forensic technique for detecting human chorionic gonadotropin hormone in pregnant women. In the forty sample batch that was dated for six months, 37 samples (92.5%) yielded positive results, 38 of which (95%) yielded good outcomes in the qualitative analysis, most of them within the sensitivities boundary. The samples that were diluted to 1/100 and 1/200 and kept at room temperature for one week and six months period, respectively, produced just two (5.0%) and five (12.5%) successful results and the samples showed negative results when diluted to 1/200 and stored for six months. Practical implication This paper reports on human chorionic gonadotropin (HCG) detection in bloodstains based on enzyme immunoassay. Conclusion: Enzyme immunoassay has proven to be a suitable method intended for detecting an hCG hormone in blood stains, allowing for the qualitative assessment of hCG, and making it particularly interesting for use in forensic science applications. Keywords: Enzyme immunoassay, human chorionic gonadotropin (hCG), pregnancy, bloodstain
Rozprawy doktorskie na temat "Blood samples and stains"
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Hwang, Suk-Moon. "Assessments of blood flow in portwine stains by laser Doppler flowmetry". Thesis, University of Strathclyde, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.366897.
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Abu-Rabie, Paul. "Direct analysis of dried blood spot samples". Thesis, University of Greenwich, 2015. http://gala.gre.ac.uk/18203/.
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The aim of the research reported herein was to identify and develop a dried blood spot (DBS) direct analysis technique that could support high sample throughput quantitative bioanalysis in a regulated drug development environment. An initial literature review, coupled with proof of concept testing of the most prominent direct analysis techniques coupled to mass spectrometers (MS), resulted in direct elution (direct extraction of DBS via a confined solvent, producing a liquid extract) being selected as the most suitable technique to develop for this application. Direct elution technology was then developed into fully automated techniques with sufficient functionality to enable compatibility with high sample throughput quantitative bioanalysis. Proof of concept robustness data demonstrated that direct elution, despite the lack of sample clean up, was a reliable technique which had no detrimental effects on detector or chromatographic performance compared to conventional wet plasma extraction and analysis. A proof of concept investigation also demonstrated that a method of improving internal standard (IS) performance by spraying IS solution onto DBS samples prior to extraction, allowed the analyte of interest and IS to be co-extracted, while retaining adequate analytical performance. The foregoing proof of concept data was then combined to produce a fully automated DBS direct elution instrument designed to introduce sample extracts into a LC-MS/MS system. This instrument incorporated a 500 DBS card capacity, an intelligent visual recognition system, a dynamic IS applicator module, and a highly effective wash system that virtually eliminates carryover. Ultimately, this work led to the production of a fully automated DBS direct elution system that is now commercially available. Subsequent research focused on optimising the system, and using this technology to address some of the issues that are currently inhibiting the development of DBS usage in drug development applications, namely haematocrit (HCT) based assay bias, and the decreased sensitivity on offer from DBS sampling. It was demonstrated that using the IS sprayer enabled the IS to integrate sufficiently with the DBS sample prior to extraction to nullify HCT based recovery bias. The direct elution mechanism was also optimised with a view to maximising assay sensitivity while retaining acceptable analytical and chromatographic (LC-MS/MS) performance. Generic optimised direct elution conditions were developed which demonstrated that increases in assay sensitivity of up to 30 fold (compared to conventional manual extraction methods) were possible using a set of representative small molecule compounds.
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Hackett, Jeffery James. "Analysis of drugs in artificially aged blood samples". Thesis, Liverpool John Moores University, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.515391.
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Del, Valle Mendoza Juana, Caso Wilmer Silva, Valdez Carmen Tinco, Maria J. Pons, Valle Luis J. Del, Oré Verónica Casabona, Michelena Denisse Champin i in. "Diagnosis of Carrion’s Disease by Direct Blood PCR in Thin Blood Smear Negative Samples". Public Library of Science (PLoS), 2014. http://hdl.handle.net/10757/315714.
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Bartonella bacilliformis is the etiologic agent of Carrion’s disease. This disease has two well established phases, the most
relevant being the so called Oroya Fever, in which B. bacilliformis infect the erythrocytes resulting in severe anemia and
transient immunosuppression, with a high lethality in the absence of adequate antibiotic treatment. The presence of
B. bacilliformis was studied in 113 blood samples suspected of Carrion’s disease based on clinical criteria, despite the
absence of a positive thin blood smear, by two different PCR techniques (using Bartonella-specific and universal 16S rRNA
gene primers), and by bacterial culture. The specific 16S rRNA gene primers revealed the presence of 21 B. bacilliformis
and 1 Bartonella elizabethae, while universal primers showed both the presence of 3 coinfections in which a concomitant
pathogen was detected plus Bartonella, in addition to the presence of infections by other microorganisms such as
Agrobacterium or Bacillus firmus. These data support the need to implement molecular tools to diagnose Carrion’s
disease.
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Du, Preez Marlize. "A comparative study of ROTEM-EXTEM results obtained from EDTA-treated whole blood samples and Sodium Citrate-treated whole blood samples in healthy volunteers". Master's thesis, University of Cape Town, 2016. http://hdl.handle.net/11427/23399.
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Background: A number of anticoagulants are available in clinical use to preserve blood samples in liquid form until a suitable time for laboratory testing. Rotational thromboelastography is usually performed on a blood sample that has been anticoagulated with sodium citrate and then recalcified immediately prior to testing. In our institution we have had shortages of citrated Vacutainer® sample tubes. The use of a single in vitro anticoagulant promises to cut costs, simplify laboratory processes as well as limit the amount of blood drawn from patients. This together with the known problems with using citrate as an anticoagulant for viscoelastic testing (VET) prompted us to investigate the suitability of EDTA as anticoagulant for VET. Method: Blood samples from 20 healthy volunteers were divided into citrated and EDTA Vacutainer® tubes. A ROTEM EXTEM® assay was performed on each sample in both groups following the manufacturer's guidelines. Clotting time (CT), clot formation time (CFT), alpha angle (α-angle) and maximum clot firmness (MCF) results were compared. Ionised calcium concentrations were measured on each sample before and after recalcification with CaCl2 to determine if there was a significant difference in post - recalcification ionised calcium concentrations between the groups. Results: The results from the two groups were treated by Bland-Altman analysis. Apart from MCF values there was significant bias between all parameters measured in the two groups. The limits of agreement for all parameters apart from MCF were unacceptable. Conclusion: We found that ROTEM EXTEM® results from EDTA samples were not comparable to or interchangeable with those from citrated samples. The difference in results is not due to differences in ionised calcium concentration levels in the samples post-recalcification as the ionised calcium concentrations in both groups post-recalcification were adequate for coagulation. EDTA samples did show superior consistency in all parameters and may be a suitable alternative for sample preservation for VET if reference ranges can be established.
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Sidiq, Farida P. "Identification of culture-negative fungi in blood and respiratory samples". Bowling Green State University / OhioLINK, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1393517514.
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Anderson, Rachel. "A review of the techniques for the forensic investigation and differentiation of human blood and decomposition fluid stains". Thesis, Anderson, Rachel (2016) A review of the techniques for the forensic investigation and differentiation of human blood and decomposition fluid stains. Masters by Coursework thesis, Murdoch University, 2016. https://researchrepository.murdoch.edu.au/id/eprint/35107/.
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An important aspect of forensic science is the detection and identification of biological fluids at a crime scene (Virkler and Lednev 2009). The determination of the type and origin of a biological sample can yield valuable information that supports a link between the criminal act and donor, which in turn may assist in the reconstruction and sequencing of a crime scene (An et al. 2012). A body and therefore any associated biological stains may not be located for a period of time, during which the decedent will begin to decompose. Blood and decomposition fluid stains have been reported to be visually similar (Comstock 2014) and therefore, it is important to determine the source of the stain. The presence of blood would suggest an injury has occurred before or shortly after death, whereas decomposition fluid is produced as a part of the naturally occurring decomposition process. Several approaches including visual examination, pH measurements, presumptive testing for blood, spectroscopic techniques, the analysis of volatile organic compounds, genomics, and proteomics may provide potential methods of biological stain identification and differentiation (Harbison and Fleming 2016; Stuart 2013; Virkler and Lednev 2009). However, there are associated limitations to these methods. This dissertation reviewed the effectiveness of these methods, which then informed the development of a proof--‐of--‐ concept study to assess if the technique of microfluidic proteomics by protein electrophoresis can identify potential differences between blood and decomposition fluid stains. When compared to conventional techniques, microfluidic devices offer many advantages including improved efficiency, a decrease in sample and reagent consumption, and automation (Li 2015). The potential results obtained from the proposed study design will assist in enhancing the knowledge base surrounding the differentiation of blood and decomposition fluid stains.
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Wellington, Emily. "Effects of Different Haematocrit Values on Estimation of Time since Deposition of Human Blood Stains Using Diffuse Reflectance Spectroscopy". Thesis, Wellington, Emily (2017) Effects of Different Haematocrit Values on Estimation of Time since Deposition of Human Blood Stains Using Diffuse Reflectance Spectroscopy. Masters by Coursework thesis, Murdoch University, 2017. https://researchrepository.murdoch.edu.au/id/eprint/37794/.
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Blood stains are a relatively common occurrence at scenes of violent crime and are a source of much information including biological, circumstantial, positional and potentially, information on the timeline of the crime. Successfully aging blood on scene could aid in time of death estimation, time since incident or even verify alibies of potential suspects. Methods in use thus far for time since death currently hold high error rates due to fluctuations in environmental factors. Several different techniques for aging of blood stains have been analysed and compared for suitability as well as similar spectroscopic techniques. Some of these aging methods were found to be complimentary, as reflectance spectroscopy has a relatively low error rate for short term aging, followed by a gradual increase, making it unsuitable for long term aging. Whereas, RNA marker analysis which follows the general degradation pattern of select RNA, was able to show a more steady error rate making it a more viable method for long term aging but not well suited to short term. The major variable factors in determining the time since deposition of blood stains include; daily or person to person fluctuations in blood protein percentages, haematocrit values, drugs present and environmental considerations such as temperature, humidity and UV light exposure. In order for any method to be acceptable in court, the full extent to these factors needs determination and full error analysis implemented. Additionally, improvement in portability, efficiency and development of non-destructive methods should be prioritised.
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Hagardson, Karin. "Comparison of DNA isolation methods to detect Leishmania parasites in blood samples". Thesis, Uppsala University, Department of Medical Biochemistry and Microbiology, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7014.
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Leishmaniasis is a disease affecting more than 12 million people worldwide. It is caused by the protozoan parasite Leishmania, which is transmitted to humans and dog hosts through bites of infected sand flies belonging to genus Phlebotomine. Several studies have shown Polymerase Chain Reaction (PCR) to be effective for the diagnosis of VL in clinical samples compared to the classical methods. The aims of this study were first to compare four different sample preparation methods for the PCR diagnosis of visceral leishmaniasis (VL) using peripheral blood samples and furthermore to find a method that is sensitive, rapid, cost benefit, simple and easy to perform. Two preparation methods were compared for the isolation of leukocytes (with Ficoll and Tris –EDTA buffer) and two DNA isolation methods (with Proteinase K and QIAgen kit). From the methods that were compared, lysis of erythrocytes with TE and the QIAgen kit seems to be the most suitable to use.
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Dinh, Louie. "Estimating cell type proportions in human cord blood samples from DNAm arrays". Thesis, University of British Columbia, 2017. http://hdl.handle.net/2429/63224.
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Epigenome-wide association studies are used to link patterns in the epigenome to human phenotypes and disease. These studies continue to increase in num- ber, driven by improving technologies and decreasing costs. However, results from population-scale association studies are often difficult to interpret. One major chal- lenge to interpretation is separating biologically relevant epigenetic changes from changes to the underlying cell type composition. This thesis focuses on computa- tional methods for correcting cell type composition in epigenome-wide association studies measuring DNAm in blood. Specifically, we focus on a class of methods, called reference-based methods, that rely on measurements of DNAm from puri- fied constituent cell types. Currently, reference-based correction methods perform poorly on human cord blood. This is unusual because adult blood, a closely related tissue, is a case-study in successful computational correction. Several previous attempts at improving cord blood estimation were only partially successful. We demonstrate how reference-based estimation methods that rely on for cord blood can be improved. First, we validated that existing methods perform poorly on cord blood, especially in minor cell types. Then, we demonstrated how this low per- formance stems from missing cell type references, data normalization and violated assumptions in signature construction. Resolving these issues improved estimates in a validation set with experimentally generated ground truth. Finally, we com- pared our reference-based estimates against reference-free techniques, an alterna- tive class of computational correction methods. Going forward, this thesis provides a template for extending reference-based estimation to other heterogeneous tissues.Science, Faculty of
Computer Science, Department of
Graduate
Książki na temat "Blood samples and stains"
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Copyright Paperback Collection (Library of Congress), red. Blood stains. New York: Kensington Pub. Corp., 2002.
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Blood stains. Waterville, Me: Thorndike Press, 2011.
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Loewen, Arley. Blood stains: A novel Afghan story. Kabul: Rahmat Publications, 2013.
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Blood stains on the veils of the kaaba. [Cairo]: State Publishing House, 1996.
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Miller, Tobin L. The use of search warrants to obtain blood samples from juveniles in "drunk driving" cases. Lansing, MI (P.O. Box 30205 Lansing 48909): Michigan Judicial Institute, 2000.
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Institute of Medicine (U.S.). Roundtable on Translating Genomic-Based Research for Health, red. Challenges and opportunities in using residual newborn screening samples for translational research: Workshop summary. Washington, D.C: National Academies Press, 2010.
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F, Siems David, i Geological Survey (U.S.), red. Analyses and descriptions of geochemical samples, Tray Mountain Wilderness, Chattachoochee Roadless Area, and Blood Mountain Wilderness, northeastern Georgia. [Reston, Va.?]: U.S. Dept. of the Interior, Geological Survey, 1988.
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Hunter, Connie. Blood Stains. Lulu.com, 2019.
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Blood Stains. MIRA, 2011.
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Blood Stains. Harlequin Enterprises, Limited, 2011.
Znajdź pełny tekst źródłaCzęści książek na temat "Blood samples and stains"
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Nielsen, Rasmus Philip. "Blood Samples". W Management of Severe Traumatic Brain Injury, 69–72. Berlin, Heidelberg: Springer Berlin Heidelberg, 2012. http://dx.doi.org/10.1007/978-3-642-28126-6_14.
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Bellander, Bo-Michael, i Rasmus Philip Nielsen. "Blood Samples". W Management of Severe Traumatic Brain Injury, 129–33. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-39383-0_19.
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Chace, Donald H., i Nicholas T. Lappas. "The Use of Dried Blood Spots and Stains in Forensic Science". W Dried Blood Spots, 140–50. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2014. http://dx.doi.org/10.1002/9781118890837.ch11.
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Brousseau, Pauline, Yves Payette, Helen Tryphonas, Barry Blakley, Herman Boermans, Denis Flipo, Michel Fournier i in. "Collection of Peripheral Blood Samples". W Manual of Immunological Methods, 7–11. Boca Raton: CRC Press, 2021. http://dx.doi.org/10.1201/9780429156977-2.
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Abu-Rabie, Paul. "Direct Analysis of Dried Blood Spot Samples". W Dried Blood Spots, 243–97. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2014. http://dx.doi.org/10.1002/9781118890837.ch22.
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Wegener, R., i J. Rummel. "Bf Typing in Blood Stains by Isoelectric Focusing". W Advances in Forensic Haemogenetics, 405–8. Berlin, Heidelberg: Springer Berlin Heidelberg, 1986. http://dx.doi.org/10.1007/978-3-642-71150-3_88.
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Spinella, A., R. Biondo, L. Garofano, A. Quaresima i D. Pellegrini. "PCR Amplification of DNA from Old Blood Stains". W Advances in Forensic Haemogenetics, 302–3. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-78782-9_78.
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Tiligada, Ekaterini, Maria Kakolyri i Madeleine Ennis. "Histamine Quantification in Human Blood Samples". W Methods in Pharmacology and Toxicology, 489–508. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-6843-5_17.
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Hannon, W. Harry, i Bradford L. Therrell. "Overview of the History and Applications of Dried Blood Samples". W Dried Blood Spots, 1–15. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2014. http://dx.doi.org/10.1002/9781118890837.ch1.
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Tutsch-Bauer, E., G. M. Weichhold i E. Josephi. "Blood Group Typing and PCR-Analysis in Stored Blood Samples". W Advances in Forensic Haemogenetics, 304–6. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-78782-9_79.
Pełny tekst źródłaStreszczenia konferencji na temat "Blood samples and stains"
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Zeng, Hansong, i Yi Zhao. "Study of Whole Blood Viscosity Using a Microfluidic Device". W ASME 2008 International Mechanical Engineering Congress and Exposition. ASMEDC, 2008. http://dx.doi.org/10.1115/imece2008-67855.
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Cardiovascular diseases include a wide range of disorders that affect heart and blood vessels, and are the leading cause of death in the United States. Whole blood viscosity, a parameter to describe the rheologic properties of blood, is an important measure of various cardiovascular diseases. It is used clinically to assess the risks of heart attack, hypertension, thrombosis and strokes. Currently used viscometers measure whole blood viscosity by inducing Couette flow to drive the blood at a certain shear rate. The blood viscosity is derived from the resistance toque measured by the toque sensor integrated within the shaft. Although effective, this method is limited due to the expensive toque sensor and the relatively large amount of blood required. More important, the fluidic conditions within the viscometer are vastly different from those in natural blood vessels (Poiseuille flow), which makes this method inappropriate to predict actual blood viscosity and its effect under natural conditions. In this work, we demonstrate whole blood viscosity measurement from the electrical resistance of the blood sample using a microfluidic device. Since the predominant parameters of the blood viscosity also determine the electrical impedance of the blood sample, the microdevice can be used as a new route of measure for blood viscosity. Blood samples with different hematocrit levels were flowed through a microchannel at different velocities that correspond to different shear rates. The electrical resistance at 20 kHz AC stimulation was recorded and compared with the viscosities measured by a commercialized rheometer. The results showed that the representative rheologic parameters (hematocrit and shear rate) are measurable by the electrical impedance. The correlation between the blood viscosity and the electrical resistance was quantitatively determined by regression analysis with a high determination coefficient. This study provides a solution for low cost, quick measurement of blood viscosity with minimal blood consumption. It also enables the in-depth investigation of blood rheology under in vivo like conditions.
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Dintenfass, L. "AGGREGATES OF RED BLOOD CELLS, AND AGGREGATES OF PLATELETS UNDER ZERO GRAVITY: EXPERIMENT ON NASA SPACE SHUTTLE "DISCOVERY" STS 51-C, JANUARY l985". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644538.
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The aim of experiment "ARC" on the space shuttle "Discovery STS 51-C, was to define effect of zero gravity on kinetics and morphology of aggregation of red cells in blood obtained from patients suffering from ischaemic heart disease, colon cancer, insulin-dependent diabetes, hyperlipidaemia, IgG and IgM papa-proteins. Space-rated automated slit-capillary photo-viscometer contained a motorized infusion pump capable of handling eight different blood samples. Two cameras and a microscope allowed micro and macrophotography, and total of 500 photographs was obtained in space; and equivalent number on the ground, in the Kennedy Space Center, where a duplicate ground photo-viscometer was present. Identical blood samples have been used in the ground experiments. The slit had a gap of 12.5 microns (micrometers). Blood was anticoagulated with EDTA and adjusted to haematocrit of 0.30 using native plasma. Samples were kept at -5°C prior to the experiment, and at 25°C during experiment; duration of experiment was 91/2 hours. The same computer program was used in both instruments. Photography was carried out at set intervals up to six minutes from the moment of stasis. There was a drastic difference between aggregation on the ground and at zero gravity. Blood from patients was greatly sludged on the ground, but normal rouleaux were formed under zero gravity. Also, aggregates uikder zero g were much smaller. However, red cell shape was not changed. Blood samples from normal donors, which showed normal rouleaux on the ground, exhibited random swarm pattern under zero gravity. Platelets, which tended to aggregate on the ground, and tended to accummulate at the slit entrance, remained monodisperse under zero gravity and no pseudopodia have been noted; under zero g platelet moved through the slit. Subject to future confirmation, it is suggested that zero gravity affects cell-to-cell interaction, and probably causes a modification of the cell membrane. If this is true, a new vista opens in the studies of immunology and oncology under zero gravity.
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Korolevich, A. N., i N. P. Prigun. "Possibilities of Lasers Optical Methods for Diagnostics of Morphological And Optical Parameters of Whole Blood Under Normal and Pathological State". W The European Conference on Lasers and Electro-Optics. Washington, D.C.: Optica Publishing Group, 1996. http://dx.doi.org/10.1364/cleo_europe.1996.cthi71.
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Sizes and number of blood aggregates of a person are known to change considerably under different diseases, such as atherosclerosis, dilates, et cetera. This works studies the possibility to use dynamical spectroscopy methods for determining sizes and their variations under normal and pathological states. Samples of whole blood were in vitro investigated for ills with different diagnoses and extends of cardio-vascular diseases (strong aggregation) as well as for essentially healthy donors.
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Spannagl, M., G. Valet i W. Schramm. "VARIATION OF FUNCTIONAL PLATELET PARAMETERS DURING STORAGE AND DURING VENOUS OCCLUSSION MEASURED". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644569.
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Information on platelet function would be of great importance for many clinical situations in addition to platelet count and bleeding time. It was the purpose of this study to test Di0c6 (3,3-dihexyl-oxacarbocyanine: transmembrane potential), AO (acridine orange: granular content), and ADB (1,4-di-acetoxy-2,3-dicyanobenzene: intracellular esterase-activity and pH) stained platelets after 1 and 5 hours storage (anticoagulated with EDTA, Heparin and Sodium-Citrate) and 2, 6, 12 and 20 minutes after venous occlussion (immediately diluted in HEPES-Buffer (1:50)). A fresh whole blood sample diluted in Buffer served as control. All blood samples were gained from normal persons. Platelets were finally diluted 1:200 in HEPES buffered saline and stained. Cell volume, green and blue fluorescence were then measured in a Fluovo-Metricell-II flow cytometer.- The mean platelet volume increased to 111% (1 h) and 117% (5 h) of control during storage. The volume remained stable during venous occlussion. - The transmembrane potential (Di0c6) decreased to 52% of control after 5 hours storage. We saw an increase to 141% after 20 minutes venous occlussion.- The granular content (A0) decreased to 81% of control during storage. There was no variation during VOT.- Esterase activity (ADB) remained constant during storage and had the lowest coefficient of variation (CV = 51%). There was an increase to 132% after 20 minutes venous occlussion.- We saw most increase in volume and decrease in Di0c6 and A0 dye content after storage in EDTA compared to Citrate and Heparin.The present results show that the dyes of functional platelet parameters are sensitively picked up by flow cytometry. The methodology seems attractive for clinical purposes because measurements can be performed in diluted blood samples within less than five minutes after venipuncture.
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"Prevalence of Hepatitis B virus and Haemoparasites among Apparently Healthy Individuals in College of Health Sciences Ladoke Akinola University of Technology Ogbomoso". W International Conference on Public Health and Humanitarian Action. International Federation of Medical Students' Associations - Jordan, 2022. http://dx.doi.org/10.56950/fynm6569.
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Hepatitis B virus (HBV) infection, a viral disease, is of great concern to health community due to its adverse effects on the liver of infected individuals. Haemoparasites are blood-dwelling parasites whose effects span from mild to severe infections. This study focused on Plasmodium falciparum Trypanosoma brucei gambiense and microfilaria which causes malaria, sleeping sickness and microfilaramia in humans respectively. This is a retrospective study that was designed to determine the prevalence of hepatitis B infection and haemoparasistes among apparently healthy individuals in the College of Health Sciences, Ladoke Akintola University of technology, Ogbomoso. Paucity of data regarding prevalence of HBV and haemoparasites among apparently healthy individuals in Ogbomoso necessitated this study. A total number of one hundred and fifty five (155) blood samples were collected within 3months for this study. Out of the one hundred and fifty five (155) blood samples collected, ten (10) tested positive to HBsAg giving a prevalence rate of 6.5%. The samples were also examined for haemoparasites on thin and thick blood smears stained with Giemsa dye using oil immersion (100X) objective of the light microscope. Only one type of haemoparasite was detected: malaria parasite with a prevalence of 87.1%. Prevalence rate for HBV and malaria parasite with respect to age group was found to be higher in age group of 25-30 and in term of sex, males have higher prevalence rate than females. The prevalence of 7.4% for co-infection of HBV and malaria parasite within the study population confirmed the high endemicity of both infections in the studied area being an urban area. It could be recommended that the Nigerian government HBV vaccination program should be extended to the adult population and not just limited to the national childhood immunization program. This is important because none of the subjects that participated in the study were vaccinated.
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Sethson, Magnus. "Extremal Optimisation Approach to Component Placement in Blood Analysis Equipment". W SICFP’21 The 17:th Scandinavian International Conference on Fluid Power. Linköping University Electronic Press, 2021. http://dx.doi.org/10.3384/ecp182p332.
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This reports present an initial study on generative mechatronic design of equipment for blood analysis where the samples and chemicals are forwarded in thin single millimeter vessels. The system of vessels in the equipment transfers the fluids to different stations where chemical reactions and studies are performed. One of the stations is an optical inspection that requires controllable lighting conditions using an array of LEDs of different types. The focus is on the generative design of the placement and configuration of the LEDs. The placement of the LEDs has been taken as a studying case for the method of Extremal Optimisation (EO) approach to mechatronic design. This method forms an opposing strategy to methods like genetic algorithms and simulated annealing. This is because it discriminate the individual parts or components of the configuration that underperform in a particular aspect instead of the more classical strategy of favouring good configurations from global measures. The presented study also relates to the class of many-objective optimisation methods (MaOP) and originates from the concept of self-organised criticality (SOC). The characteristics of avalanche barrier crossings in the parameter search space is inherited from such systems. The test case used for the evaluation places occupying circles onto a quarter ring domain representing LEDs and circuit board. The fluid vessels are represented by lit up small domains that are also approximated by a circular disc. Some conclusion upon the methods capability to form a valid solution are made. A framework for describing a set of local flaw-improvement rules, called D2FI is introduced.
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Voeikov, Vladimir, Ekaterina Buravleva, Yulia Bulargina i Yuri Gurfinkel. "Diagnostic applications of an opto-electronic device for high temporal resolution of erythrocytes sedimentation (ESR-graphy)". W European Conference on Biomedical Optics. Washington, D.C.: Optica Publishing Group, 2001. http://dx.doi.org/10.1364/ecbo.2001.4434_208.
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An automatic device for high-temporal resolution of the process of erythrocytes sedimentation in blood was designed. The position of the boundary between red blood and plasma is registered each 30 sec in several pipettes simultaneously with ±10 mkm precision. Data are processed by a PC and presented as velocity-time curves (ESR-grams) and the curves describing time evolution of the boundary position. ESR-grams demonstrate non-monotonous character of erythrocytes sedimentation in blood. Blood of particular donor being in a stable physiological state taken on different days is characterized by similar ESR-grams. Pathological deviations from a normal physiological state are reflected in the shortening of duration of each process stage and increasing of average sedimentation rate. Intravenous infusion of some medical preparations may lead either to improving (prolonging of macrokinetic stages, decreasing of sedimentation rate), or to worsening of studied parameters depending on an individual. The low extent of blood dilution with saline in vitro lead as a rule to decreasing of sedimentation rate and improving of microkinetic parameters of the process. Adding of highly diluted hydrogen peroxide to blood samples of patients resulted in the improving of sedimentation kinetics. ESR-graphy may widen opportunities of practical medicine in diagnostics, prognostics and drug therapy.
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Goodfield, M. J. D., M. A. Orchard i N. R. Rowell. "REDUCED THRESHOLD FOR COLLAGEN INDUCED PLATELET AGGREGATION IN WHOLE BLOOD FROM PATIENTS WITH SYSTEMIC SCLEROSIS". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643478.
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Systemic sclerosis is a multi-system disorder characterised by skin lesions and narrowing of arterioles with reduction in blood supply to many organs. Previous reports have suggested that platelet aggregation is abnormal and could contribute to the pathogenesis. Platelet aggregation in whole blood was investigated in 10 patients with systemic sclerosis and 9 age matched controls. Subjects rested for 30 minutes before blood samples were taken without stasis, via a 19G needle into 3.8% trisodium citrate anti-coagulant. 0.5ml of blood was transferred to cuvettes, and equilibrated gt 22°C for 30 minutes, before being stirred at 750 r.p.m. at 37 C. Aggregation was induced by collagen (0.01 ug/1 to 1.0 ug/1), adrenaline (0,1,0.5 and 1.0 ug/1) and ADP (0.1,0.25 and 2.5 ug/1). Aliquots for counting free platelets were taken and assessed using the Ultraflow 100 particle counter.Platelet aggregation was significantly increased in patients with systemic sclerosis at all doses of collagen (p<0.01), and the mean dose giving 50% aggregation was significantly lower in the disease group (0.026+/-0.06mg/l) than in the controls (0.23+/-0.06mg/l, p<0.01). Aggregation was not significantly different with the other aggregating agents.Increased sensitivity of platelets to aggregation by collagen in whole blood may be of importance in the pathogenesis of the vascular and fibrotic lesions of this disease
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Kuula, J., H. H. Puupponen, H. Rinta i I. Pölönen. "The challenges of analysing blood stains with hyperspectral imaging". W SPIE Sensing Technology + Applications, redaktorzy Šárka O. Southern, Mark A. Mentzer, Isaac Rodriguez-Chavez i Virginia E. Wotring. SPIE, 2014. http://dx.doi.org/10.1117/12.2050180.
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Bolyahina, S. A., i E. A. Efremova. "COMPARISON OF MORPHOLOGICAL CHANGES IN THE BLOOD OF HENS DURING EXPERIMENTAL INFECTION WITH T. SPIRALIS AND T. PSEUDOSPIRALIS". W THEORY AND PRACTICE OF PARASITIC DISEASE CONTROL. All-Russian Scientific Research Institute for Fundamental and Applied Parasitology of Animals and Plant – a branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV”, 2023. http://dx.doi.org/10.31016/978-5-6048555-6-0.2023.24.95-99.
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A comparison was done for changes in the morphological composition of blood in
hens experimentally infected with T. pseudospiralis and T. spiralis larvae at a dose
of 2 larvae/g m.a., intragastrically. It was established that trichinellosis in the hens
due to the parasitism of a non-encapsulated Trichinella species caused regenerative
hypochromic anemia. Indicators of the trichinellosis process are granulocytic series
cells, heterophils. The onset of the body's reaction to infection was detected on
day 4 after invasion and was expressed by mild neutrophilia. Then an increase was
observed in the content of heterophils in the blood of the birds with a maximum on
day 29, which corresponds to the migration and muscular stage of the trichinellosis
process. Subsequently, there is a relative and absolute quantitative decrease in
these groups of cells in the bloodstream of the poultry, however, these indicators
are higher during the entire observation period (61 days) than in the hens of the
control group. Lymphocytes in the blood samples from the experimental hens have
an absolute increase on day 50 from the infection, which can be characterized as an
immunological reaction of the hens in the invasion. The dynamics of the level of
eosinophils and basophils circulating in the blood of the poultry is due to the stages
of the trichinellosis process and confirms the predominance of an allergic reaction in the nature of the disease. However, more pronounced quantitative changes in
hematological parameters, especially on the part of heterophils and eosinophils were
recorded in the poultry infected with a non-encapsulated Trichinella species.
Raporty organizacyjne na temat "Blood samples and stains"
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Shpigel, Nahum, Raul Barletta, Ilan Rosenshine i Marcelo Chaffer. Identification and characterization of Mycobacterium paratuberculosis virulence genes expressed in vivo by negative selection. United States Department of Agriculture, styczeń 2004. http://dx.doi.org/10.32747/2004.7696510.bard.
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Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of a severe inflammatory bowel disease (IBD) in ruminants, known as Johne’s disease or paratuberculosis. Johne’s disease is considered to be one of the most serious diseases affecting dairy cattle both in Israel and worldwide. Heavy economic losses are incurred by dairy farmers due to the severe effect of subclinical infection on milk production, fertility, lower disease resistance and early culling. Its influence in the United States alone is staggering, causing an estimated loss of $1.5 billion to the agriculture industry every year. Isolation of MAP from intestinal tissue and blood of Crohn's patients has lead to concern that it plays a potential pathogenic role in promoting human IDB including Crohn’s disease. There is great concern following the identification of the organism in animal products and shedding of the organism to the environment by subclinically infected animals. Little is known about the molecular basis for MAP virulence. The goal of the original proposed research was to identify MAP genes that are required for the critical stage of initial infection and colonization of ruminants’ intestine by MAP. We proposed to develop and use signature tag mutagenesis (STM) screen to find MAP genes that are specifically required for survival in ruminants upon experimental infection. This research projected was approved as one-year feasibility study to prove the ability of the research team to establish the animal model for mutant screening and alternative in-vitro cell systems. In Israel, neonatal goat kids were repeatedly inoculated with either one of the following organisms; MAP K-10 strain and three transposon mutants of K-10 which were produced and screened by the US PI. Six months after the commencement of inoculation we have necropsied the goats and taken multiple tissue samples from the jejunum, ileum and mesenteric lymph nodes. Both PCR and histopathology analysis indicated on efficient MAP colonization of all the inoculated animals. We have established several systems in the Israeli PI’s laboratory; these include using IS900 PCR for the identification of MAP and using HSP65-based PCR for the differentiation between MAV and MAP. We used Southern blot analysis for the differentiation among transposon mutants of K-10. In addition the Israeli PI has set up a panel of in-vitro screening systems for MAP mutants. These include assays to test adhesion, phagocytosis and survival of MAP to/within macrophages, assays that determine the rate of MAPinduced apoptosis of macrophages and MAP-induced NO production by macrophages, and assays testing the interference with T cell ã Interferon production and T cell proliferation by MAP infected macrophages (macrophage studies were done in BoMac and RAW cell lines, mouse peritoneal macrophages and bovine peripheral blood monocytes derived macrophages, respectively). All partners involved in this project feel that we are currently on track with this novel, highly challenging and ambitious research project. We have managed to establish the above described research systems that will clearly enable us to achieve the original proposed scientific objectives. We have proven ourselves as excellent collaborative groups with very high levels of complementary expertise. The Israeli groups were very fortunate to work with the US group and in a very short time period to master numerous techniques in the field of Mycobacterium research. The Israeli group has proven its ability to run this complicated animal model. This research, if continued, may elucidate new and basic aspects related to the pathogenesis MAP. In addition the work may identify new targets for vaccine and drug development. Considering the possibility that MAP might be a cause of human Crohn’s disease, better understanding of virulence mechanisms of this organism might also be of public health interest as well.
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Udey, R., T. Corzett i A. Williams. Extraction of Butyrylcholinesterase (BuChE) and Organophosphorus Nerve Agent (OPNA)-BChE Adducts from Blood and Plasma Samples and Analysis by Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS). Office of Scientific and Technical Information (OSTI), luty 2014. http://dx.doi.org/10.2172/1149044.
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Glazer, Itamar, Alice Churchill, Galina Gindin i Michael Samish. Genomic and Organismal Studies to Elucidate the Mechanisms of Infectivity of Entomopathogenic Fungi to Ticks. United States Department of Agriculture, styczeń 2013. http://dx.doi.org/10.32747/2013.7593382.bard.
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The overall goal of this research was to elucidate the factors affecting early development of Metarhizium spp. (previously named M. anisopliae) on ticks or tick cuticle extracts and the molecular basis of these early infection processes. The original objectives were: 1. Characterize the pre-penetration events (adhesion, germination and appressorium formation) of spores of M. anisopliae strains with high or low virulence during tick infection. 2. Create GFP-expressing strains of M. anisopliae tick pathogens having high and low virulence to compare their progress of infection by microscopy. 3. Use microarray analyses, primarily with existing M. anisopliae EST sequences in GenBank, to identify and characterize fungal genes whose expression is regulated in response to host cuticle extracts. Objective 3 was later modified (as approved by BARD) to use RNAseq to characterize the early stages of fungal gene expression during infection of intact host cuticles. This new method provides a massively larger and more informative dataset and allows us to take advantage of a) recently published genomes of Metarhizium robertsii and M. acridum for RNAseq data analysis, and b) newly developed and highly efficient cDNA sequencing technologies that are relatively low cost and, therefore, allow deep sequencing of multiple transcriptome samples. We examined pre-penetration and penetration events that differentiate high and low virulence strains of Metarhizium spp., focusing on spore adhesion, germination, appressorium formation, and penetration of tick integuments. Initiation of fungal infection was compared on susceptible and resistant tick species at different tick developmental stages. In vitro studies comparing the effects of protein and fatty acid profiles from tick cuticle extracts demonstrated that resistant tick cuticles contain higher concentrations of specific lipids that inhibit fungal development than do susceptible tick cuticles, suggesting one mechanism of Ixodidae resistance to fungal entomopathogens (Objective 1). We used molecular markers to determine that the three M. anisopliae strains from Israel that we studied actually were three distinct species. M. brunneum is highly virulent against the tick Rhipicephalus annulatus, M. pingshaense and M. robertsii are intermediate in virulence, and M. majus is of low virulence. We transformed all four Metarhizium species to express GFP and used them in pathogenicity assays against diverse tick species. Key findings were that a) resistant ticks inhibit Metarhizium infection prior to hemocoel invasion by reducing fungal viability on the cuticle surface (Objective 2), as was supported by the in vitro studies of Objective 1, and b) Metarhizium kills susceptible ticks after cuticle penetration but prior to hemocoel colonization. Transcriptome studies of the most virulent species, M. brunneum, are in progress and include analyses of ungerminated conidia and conidia germination and development on a low nutrient medium or on susceptible R. annulatus exoskeleton (Objective 3). We anticipate these studies will contribute to identifying fungal genetic factors that increase virulence and speed of kill and may help reveal tick chemistries that could be included in biocontrol formulations to increase efficacy. Methodologies developed to screen tick cuticle extracts for ability to support conidia germination and development may help in the selection of wild fungi with increased virulence against resistant ticks. The overall knowledge gained should contribute not only to the improvement of tick control but also to the control of other blood-sucking arthropods and related plant pests. Use of bio-based agents for controlling arthropods will contribute to a healthier, more sustainable environment and serve a growing number of organic food farmers.
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Peng, Ciyan, Jing Chen, Sini Li i Jianhe Li. Comparative Efficacy of Chinese Herbal Injections Combined Western medicine for Non-small cell lung cancer: A Bayesian Network Meta-Analysis of randomized controlled trials. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, listopad 2021. http://dx.doi.org/10.37766/inplasy2021.11.0068.
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Review question / Objective: Advanced lung cancer has become the top malignant tumor in terms of morbidity and mortality, and Chinese herbal injections combined with western drugs have been widely used to treat advanced non-small cell lung cancer. For this purpose, we conducted a Bayesian network analysis to systematically evaluate the efficacy of different herbal injections combined with western drugs in the treatment of NSCLC. Subjects: Patients diagnosed with NSCLC by pathological or cytological examination, locally advanced or those who refused surgical treatment were included, regardless of gender, age, stage, race, nationality and sample size; Interventions: Chinese herbal injections combined with three types of commonly used western drugs (platinum, targeted and immune agents) were used in the experimental group, while the control group was treated with western drugs alone; Study type: to report the efficacy of Chinese herbal injections combined with western drugs in the treatment of non-small cell lung cancer efficacy in a randomized controlled trial (rct) Eligible. No restrictions were imposed on language, year of publication, or publication status. Ending indicators: Main ending indicators: (1) disease control rate (DCR), DCR = (complete remission + partial remission + stable)/total number of cases. Efficacy rate = (number of improvement cases + number of stable cases)/total number of cases. (2) Secondary outcome indicators: quality of life, determined according to the KPS behavioral status scale, improvement was defined as an increase of ≥10 points in KPS score after treatment; stability was defined as an increase or decrease of <10 points in KPS score; decline was defined as a decrease of ≥10 points in KPS score. (3) The incidence of adverse reactions, including gastrointestinal reactions, white blood cell (WBC) reduction, hemoglobin (HGB) reduction, platelet (PLT) reduction, etc.
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Cahaner, Avigdor, Sacit F. Bilgili, Orna Halevy, Roger J. Lien i Kellye S. Joiner. effects of enhanced hypertrophy, reduced oxygen supply and heat load on breast meat yield and quality in broilers. United States Department of Agriculture, listopad 2014. http://dx.doi.org/10.32747/2014.7699855.bard.
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Original objectivesThe objectives of this project were to evaluate the growth performance, meat yield and quality attributes of broiler strains widely differing in their genetic potential under normal temperature vs. warm temperature (short and long-term) conditions. Strain differences in breast muscle accretion rate, metabolic responses under heat load and, gross and histopathological changes in breast muscle under thermal load was also to be characterized. BackgroundTremendous genetic progress has been made in broiler chicken growth rate and meat yield since the 1950s. Higher growth rate is driven by higher rates of feed intake and metabolism, resulting in elevated internal heat production. Hot rearing conditions negatively affect broiler growth by hindering dissipation of heat and may lead to a lethal elevation in body temperature. To avoid heat-induced mortality, broilers reduce feed intake, leading to depressed growth rate, lower weight gain, reduce breast meat yield and quality. Thus, the genetic potential of contemporary commercial broilers (CCB) is not fully expressed under hot conditions. Major conclusions, solutions, and achievementsResearch conducted in Israel focused on three broiler strains – CCB, Featherless, Feathered sibs (i.e., sharing similar genetic background). Complimentary research trials conducted at Auburn utilized CCB (Cobb 500, Cobb 700, Ross 308, Ross 708), contrasting their performance to slow growing strains. Warm rearing conditions consistently reduced feed intake, growth rate, feed efficiency, body weight uniformity and breast muscle yield, especially pronounced with CCB and magnified with age. Breast meat quality was also negatively affected, as measured by higher drip loss and paler meat color. Exposure to continuous or short-term heat stress induced respiratory alkalosis. Breast muscle histomorphometrics confirmed enhanced myofiber hypertrophy in CCB. Featherless broilers exhibited a significant increase in blood-vessel density under warm conditions. Rapid growth and muscle accretion rate was correlated to various myopathies (white striping, woody and necrotic) as well as to increases in plasma creatinekinase levels. Whether the trigger(s) of muscle damage is loss of cellular membrane integrity due to oxidative damage or tissue lactate accumulation, or to loss of inter-compartmental cation homeostasis is yet to be determined. Based on genome-wide single-nucleotide polymorphism array genotyping, identification of the gene with the recessive mutation Scaleless (sc) facilitated the development a dCAPS assay to discriminate between sc carrier (sc/+) and non-carrier (+/+) individuals. ImplicationsThis project confirmed that featherless broiler strains grow efficiently with high yield and quality of breast meat, even under warm rearing conditions that significantly depress the overall performance of CCB. Therefore, broiler meat production in hot regions and climates can be substantially improved by introducing the featherless gene into contemporary commercial broiler stocks. This approach has become more feasible with the development of dCAPS assay. A novel modification of the PCR protocol (using whole blood samples instead of extracted DNA) may contribute to the efficient development of commercial featherless broiler strains. Such strains will allow expansion of the broiler meat production in developing countries in warm climates, where energy intensive environmental control of rearing facilities are not economical and easily achievable.
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D’Agostino, Martin, Nigel Cook, Liam O’Connor, Annette Sansom, Dima Semaan, Anne Wood, Sue Keenan i Linda Scobie. Optimising extraction and RT-qPCR-based detection of hepatitis E virus (HEV) from pork meat and products. Food Standards Agency, lipiec 2023. http://dx.doi.org/10.46756/sci.fsa.ylv958.
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Hepatitis E is an infection of the liver caused by the hepatitis E virus (HEV). HEV infection usually produces a mild disease, hepatitis E. However, disease symptoms can vary from no apparent symptoms to liver failure. There are 4 main types (genotypes) of the virus that cause concern in humans. Genotypes 1 and 2 infections are mainly restricted to humans but 3 and 4 can be identified in numerous other animal species including pigs. Transmission routes of HEV genotypes 3 and 4 have been identified to include the consumption of food products derived from infected animals and shellfish, and via transfusion of infected blood products. Hepatitis E infection is still an emerging issue in the UK and there is evidence to suggest an association of this virus with undercooked pork and pork products. Currently, there is no standardized method for evaluating the stability of HEV that may be present in food during cooking processes. There is also lack of a suitable method that can detect only infectious HEV. The proposed project aimed to address a key gap in resources for methodology related to the detection of HEV in pork and pork products. Currently the lack of a standardised method for the detection of HEV has resulted in individual laboratories either utilising their own methods or adapting methods from previously published work. This leads to a high degree of variability between the interpretation of results and does nothing to progress or provide benefit to the food industry. By interrogating the existing published methods, the project sought to refine and optimise elements of existing protocols in order to enhance the performance characteristics of the method and to simplify the methodology wherever possible. The aim was to produce a validated method which is both robust and repeatable which can be easily integrated into food laboratories capable of performing virus related work. Overall, the final method chosen was devoid of hazardous reagents and utilised easily accessible equipment. To verify the robustness of the method, an international collaborative trial was performed, with 4 UK and 3 European participant laboratories. The participating laboratories conducted analyses of pork liver samples artificially contaminated with various levels of HEV (including uncontaminated samples). The trial showed that the HEV DETECT method was just as reproducible between laboratories as it was repeatable within a laboratory. It is envisaged that the developed system will be put forward as a suitable candidate for ISO certification as a standard method. The establishment of these methods in UK laboratories could result in the availability of independent testing services for both domestic and imported pork /pork-based products. The availability of this method is in essence innovation. This work is essential to industry to help support further research to ensure that public health safety and confidence in pork and other “HEV risk” food products is maintained and improved.
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Davidson, Irit, Hsing-Jien Kung i Richard L. Witter. Molecular Interactions between Herpes and Retroviruses in Dually Infected Chickens and Turkeys. United States Department of Agriculture, styczeń 2002. http://dx.doi.org/10.32747/2002.7575275.bard.
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Tumors in commercial poultry are caused mainly by infection with avian herpes and retroviruses, the herpesvirus Marek's disease virus (MDV) and the retroviruses, reticuloendotheliosis (REV), lymphoid leukosis, subgroups A-I and J (ALV and ALV-J) in chickens, or Iymphoprolipherative disease (LPDV) in turkeys. Infection with one virus aggravates the clinical outcome of birds that are already infected by another oncogenic virus. As these viruses do not interfere for infection, MDV and one or more retroviruses can infect the same flock, the same bird and the same cell. While infecting the same cell, herpes and retroviruses might interact in at least three ways: a) Integration of retrovirus genomes, or genomic fragments (mainly the LTR) into MDV;b) alteration of LTR-driven expression of retroviral genes by MDV immediate- early genes, and c) by herpesvirus induced cellular transcriptional factors. The first type of molecular interaction have been demonstrated to happen efficiently in vitro by Dr. Kung, in cases multiple infection of cell cultures with MDV and REV or MDV and ALV. Moreover, Dr. Witter showed that an in vitro-created recombinant, RM1, had altered in vitro replication and in vivo biological properties. A more comprehensive characterization of RM1 was carried out in the present project. We sought to highlight whether events of such integrations occur also in the bird, in vivo. For that, we had first to determine the prevalence of dually-infected individual birds in commercial flocks, as no systematic survey has been yet reported. Surprisingly, about 25% of the commercial flocks infected with avian oncogenic viruses had a multiple virus infection and 5% of the total samples ana lysed had multiple virus sequences. Then, we aimed to evaluate and characterize biologically and molecularly the resulting recombinants, if formed, and to analyse the factors that affect these events (virus strains, type and age of birds and time interval between the infection with both viruses). The perception of retrovirus insertions into herpesviruses in vivo is not banal, as the in vivo and in vitro systems differ in the viral-target cells, lymphocytes or fibroblasts, in the MDV-replicative type, transforming or productive, and the immune system presence. We realized that previous methods employed to study in vitro created recombinant viruses were not adequate for the study of samples taken directly from the bird. Therefore, the Hot Spot-combined PCR was developed based on the molecularly known RM1 virus. Also, the PFGE that was used for tissue cultured-MDV separation was inefficient for separating MDV from organs, but useful with feather tips as a source of bird original MDV. Much attention was dedicated now to feathers, because if a recombinant virus would be formed in vivo, its biological significance would be evident by horizontal dissemination through the feathers. Major findings were: a) not only in vitro, but also in vivo MDV and retrovirus co-infections lead to LTR integrations into MDV. That was shown by the detection of chimeric molecules. These appeared in low quantities and as quasispecies, thus interfering with sequence analysis of cloned gel-purified chimeric molecules. Mainly inserts were located in the repeat long MDV fragments. In field birds chimeric molecules were detected at a lower frequency (2.5%) than in experimentally infected birds (30-50%). These could be transmitted experimentally to another birds by inoculation with chimeric molecules containing blood. Several types of chimeric molecules were formed, and same types were detected in birds infected by a second round. To reproduce viral integrations, in vivo infection trials were done with field inoculate that contained both viruses, but the chimeric molecule yield was undetectable.
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Weinberg, Zwi G., Adegbola Adesogan, Itzhak Mizrahi, Shlomo Sela, Kwnag Jeong i Diwakar Vyas. effect of selected lactic acid bacteria on the microbial composition and on the survival of pathogens in the rumen in context with their probiotic effects on ruminants. United States Department of Agriculture, styczeń 2014. http://dx.doi.org/10.32747/2014.7598162.bard.
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This research project was performed in context of the apparent probiotic effect of selected lactic acid bacteria (LAB) silage inoculants on the performance of ruminants (improved feed intake, faster live-weight gain, higher milk yields and improved feed efficiency). The overall objective was to find out how LAB affect ruminant performance. The project included several “chapters” as follows: 1. The effect of LAB silage inoculants on the survival of detrimental bacteria in rumen fluid, in vitro study (Weinberg et al., The Volcani Center). An in vitro model was developed to study the interaction between selected LAB and an E. coli strain tagged with green fluorescence protein (GFP) in buffered RF. Results indicated that both LAB inoculants and E. coli survived in the RF for several days; both LAB inoculants and LAB-treated silages did not affect survival of E. coli in rumen fluid in vitro. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the performance of high-lactating cows (Weinberg et al., The Volcani Center). Treatments included control (no additive), Lacobacillusbuchneri40788 (LB), Lactobacillus plantarumMTD1 40027 (LP) and Pediococcuspentosaceus30168 (PP), each applied at 10⁶ cfu/g FM. The silages were included in the TMR of 32 high milking Holstein cows in a controlled feeding experiment. All baled silages were of good quality. The LB silage had the numerically highest acetic acid and were the most stable upon aerobic exposure. The cows fed the LB silages had the highest daily milk yields, percent milk fat and protein. The microbiome of baled wheat silages and changes during ensiling of wheat and corn (Sela et al., The Volcani Center). Bacterial community of the baled silages was dominated mainly of two genera in total, dominated by Lactobacillus and Clostridium_sensu_stricto_12 with 300 other genera at very low abundance. Fungal community was composed mainly of two genera in total, dominated by Candida and Monascuswith 20 other genera at very low abundance. In addition, changes in the microbiome during ensiling of wheat and corn with and without addition of L. plantarumMTD1 was studied in mini-silos. Overall 236 bacterial genera were identified in the fresh corn but after 3 months Lactobacillus outnumbered all other species by acquiring 95% of relative abundance. The wheat silage samples are still under analysis. The effect of applying LAB inoculants at ensiling on survival of E. coli O157:H7 in alfalfa and corn silages(Adesogan et al., University of Florida). E. coli (10⁵ cfu/g) was applied to fresh alfalfa and corn at ensiling with or without L. plantarumor L. buchneri. The pathogen was added again after about 3 moths at the beginning of an aerobic exposure period. The inoculants resulted in faster decrease in pH as compared with the control (no additives) or E. coli alone and therefore, the pathogen was eliminated faster from these silages. After aerobic exposure the pathogen was not detected in the LAB treated silages, whereas it was still present in the E. coli alone samples. 5. The effect of feeding corn silage treated with or without L. buchnerion shedding of E. coli O157:H7 by dairy cows (Adesogan et al., UFL). BARD Report - Project 4704 Page 2 of 12 Five hundred cows from the dairy herd of the University of Florida were screened for E. coli shedding, out of which 14 low and 13 high shedders were selected. These cows were fed a total mixed ration (TMR) which was inoculated with E. coli O157:H7 for 21 days. The TMR included corn silage treated with or without L. buchneri. The inoculated silages were more stable upon aerobic exposure than the control silages; the silage inoculant had no significant effect on any milk or cow blood parameters. However, the silage inoculant tended to reduce shedding of E. coli regardless of high or low shedders (p = 0.06). 6. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the rumen microbiome (Mizrahi et al., BGU). Rumen fluid was sampled throughout the feeding experiment in which inoculated wheat silages were included in the rations. Microbial DNA was subsequently purified from each sample and the 16S rRNA was sequenced, thus obtaining an overview of the microbiome and its dynamic changes for each experimental treatment. We observed an increase in OTU richness in the group which received the baled silage inoculated with Lactobacillus Plantarum(LP). In contrast the group fed Lactobacillus buchneri(LB) inoculated silage resulted in a significant decrease in richness. Lower OTU richness was recently associated in lactating cows with higher performance (Ben Shabatet al., 2016). No significant clustering could be observed between the different inoculation treatments and the control in non metric multi-dimentional scaling, suggesting that the effect of the treatments is not the result of an overall modulation of the microbiome composition but possibly the result of more discrete interactions. Significant phylum level changes in composition also indicates that no broad changes in taxa identity and composition occurred under any treatment A more discrete modulation could be observed in the fold change of several taxonomic groups (genus level analysis), unique to each treatment, before and after the treatment. Of particular interest is the LB treated group, in which several taxa significantly decreased in abundance. BARD Report - Project 4704 Page 3 of 12
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Fourth national report on human exposure to environmental chemicals. National Center for Environmental Health, marzec 2021. http://dx.doi.org/10.15620/cdc:105345.
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"The Updated Tables, March 2021) presents nationally representative, cumulative biomonitoring data gathered from 1999–2000 through 2015–2016. It includes all the data from each of the previous National Reports on Human Exposure to Environmental Chemicals and each of the previous Updated Tables (collectively, the Report and Updated Tables). In each survey period, the reported chemicals or their metabolites were measured in blood, serum, and urine samples from random subsamples of the National Health and Nutrition Examination Survey (NHANES). These subsamples typically consisted of about 2,500 participants – exact numbers are included in the tables. Survey data and samples are collected by the Centers for Disease Control and Prevention (CDC) National Center for Health Statistics. CDC’s Environmental Health Laboratory (Division of Laboratory Sciences (DLS), National Center for Environmental Health) used mass spectrometry methods to obtain the blood, serum, and urine exposure measurements presented in the Report and Updated Tables. Volume One (1999-2010) and Volume Two (2011-2016) contain data tables for chemicals measured in the general U.S. population Volume Two: NHANES 2011-2016 provides data on the general U.S. population from NHANES 2011-2012, 2013–2014, and 2015-2016. CS272983-A FourthReport_UpdatedTables_Volume2_Mar2021-508.pdf"
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Fourth national report on human exposure to environmental chemicals. Updated tables, March 2021 : volume two: NHANES 2011-2016. National Center for Environmental Health (U.S.), marzec 2021. http://dx.doi.org/10.15620/105345.
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"The Fourth National Report on Human Exposure to Environmental Chemicals: Updated Tables, March 2021 (the Updated Tables, March 2021) presents nationally representative, cumulative biomonitoring data gathered from 1999–2000 through 2015–2016. It includes all the data from each of the previous National Reports on Human Exposure to Environmental Chemicals and each of the previous Updated Tables (collectively, the Report and Updated Tables). In each survey period, the reported chemicals or their metabolites were measured in blood, serum, and urine samples from random subsamples of the National Health and Nutrition Examination Survey (NHANES). These subsamples typically consisted of about 2,500 participants – exact numbers are included in the tables. Survey data and samples are collected by the Centers for Disease Control and Prevention (CDC) National Center for Health Statistics. CDC’s Environmental Health Laboratory (Division of Laboratory Sciences (DLS), National Center for Environmental Health) used mass spectrometry methods to obtain the blood, serum, and urine exposure measurements presented in the Report and Updated Tables. Volume One (1999-2010) and Volume Two (2011-2016) contain data tables for chemicals measured in the general U.S. population Volume Two: NHANES 2011-2016 provides data on the general U.S. population from NHANES 2011-2012, 2013–2014, and 2015-2016. CS272983-A FourthReport_UpdatedTables_Volume2_Mar2021-508.pdf"