Samples Of Biological Material Research Articles

Cytomorphometric Analysis of Inflammation Dynamics in the Periodontium Following the Use of Fixed Dental Prostheses.

Cytomorphometry is used in the sampling of biological materials and diagnostic procedures. The use of cytological studies in periodontal diseases is not well described in the literature. Our study aimed to quantitatively assess the inflammation dynamics using cytomorphometric analysis of the periodontium before and after the use of fixed dental prostheses. Following ethics approval, a total of 105 subjects were divided in 3 groups as gingivitis (n = 23), periodontitis (n = 58), and healthy periodontium (control) (n = 24). The fixed dental prostheses (crowns and fixed partial dentures) were fabricated from cobalt-chrome metal-ceramic prostheses using the conventional method (C/M-CoCr), cobalt-chrome metal-ceramic prostheses by the computer-aided design and computer-aided manufacturing (CAD/CAM) technique (C/C-CoCr), and zirconia-based ceramic prostheses by the CAD/CAM technique (C/C-Zr) among subjects with gingivitis and periodontitis. The gingival crevicular fluid (GCF) was obtained from subjects before and after the use of the prostheses. The total count of epithelial cells and the connective tissue cells or polymorphonuclear neutrophils (PMNs) in GCF were studied using cytomorphometric analysis. The Statistical Package Tor the Social Sciences (SPSS), Version 20 (IBM Company, Chicago, IL, USA) was used to analyze the results and the significance level was set at p = 0.05. The data for before and after the use of the prostheses were compared using independent t-Tests. Similarly, the results after the use of prostheses in gingivitis, periodontitis, and control in each type of prostheses were compared using One-way ANOVA with post hoc using Scheffe. The total epithelial cells and the PMNs were determined along with the epithelium/leukocyte index. Regardless of the prostheses type used, no significant change in the parameters was identified among patients with a healthy periodontium, before and after prosthetic treatment. In all study groups, a statistically increase (p value < 0.05) was observed in the oral epithelial cell counts and a statistically decrease (p < 0.05) in the PMNs count following the use of the fixed prostheses. Data on cytomorphometric analysis could enable the selection of the most appropriate prostheses for use in patients with periodontal pathologies. When choosing prostheses, changes in the composition of GCF could be considered as a useful criterion for their use.

Liquid biopsy as a perioperative biomarker of digestive tract cancers: review of the literature.

Tissue biopsies are the gold-standard for investigating the molecular characterization of tumors. However, a "solid" biopsy is an invasive procedure that cannot capture real-time tumor dynamics and may yield inaccurate information because of intratumoral heterogeneity. In this review, we summarize the current state of knowledge about surgical treatment-associated "liquid" biopsy for patients with digestive organ tumors. A liquid biopsy is a technique involving the sampling and testing of non-solid biological materials, including blood, urine, saliva, and ascites. Previous studies have reported the potential value of blood-based biomarkers, circulating tumor cells, and cell-free nucleic acids as facilitators of cancer treatment. The applications of a liquid biopsy in a cancer treatment setting include screening and early diagnosis, prognostication, and outcome and recurrence monitoring of cancer. This technique has also been suggested as a useful tool in personalized medicine. The transition to precision medicine is still in its early stages. Soon, however, liquid biopsy is likely to form the basis of patient selection for molecular targeted therapies, predictions regarding chemotherapy sensitivity, and real-time evaluations of therapeutic effects.

Characteristics of allele pool of cattle of simmental breed in Kazakhstan

One of the key aspects of studying of new breeds, types and lines of farm animals is to evoluation the state of their allele pool by using DNA markers. At the same time, it should be noted that a significant contribution to the characteristic of allelic diversity of breeds is made by regional populations, the formation of which in most cases took place on the basis of local cattle, which are carriers of their own unique allele pool. It is known that studies on microsatellites make it possible to determine the genetic diversity within and between breeds, and the average number of alleles, observed and expected heterozygosity, allow us to calculate the genetic parameters of populations and evaluate their diversity. The purpose of the research was to study the features of the allele pool of a new intra-breed type Ertis of Red-and-White dairy cattle in the Simmental breed. The sampling of biological materials has been carried out according to generally accepted methods. Genotyping has been performed with a set of StockMarks Cattle at 12 loci. Identification of amplification products has been performed on the abi Prism 310 genetic analyzer (Applied Biosystems, USA) using capillary electrophoresis. Decoding of the obtained graphical results has been carried out in the program GeneMapper 4,0. The results of researches on the genetic structure of the studied population of cattle have been reflected in the article. It has been found that there are 65 alleles in 12 loci of the microsatellite, of which 7 alleles with a frequency of occurrence of ≤1 % have been identified. The most informative loci were TGLA227, TGLA122, ETH3, and TGLA53. The average level of expected heterozygosity was 0,68, with random inbreeding – 0,0488. Studies of the genetic profile of the new intra-breed type of dairy cattle Ertis allowed us to establish the wide variety of allele pool in it, and the small number of rare alleles allocated gives grounds for the possibility of determining this population as an independent structural unit in Simmental breed.

Water security and detection of Cryptosporidium spp. in samples from the Guarapiranga Reservoir, São Paulo

Water security depends, among other factors, on the hydrological cycle, quantity and quality of water. Several microorganisms that pose risks to human and animal health are waterborne, such as the protozoan Cryptosporidium spp. The parasitological diagnosis of Cryptosporidium spp. in biological material and environmental samples can be accomplished through several techniques that include optical microscopy with analysis of fresh material or through the use of concentration techniques, indirect immunofluorescence and molecular biology that allows differentiation between species. Oocysts of Cryptosporidium spp. are eliminated through feces from parasitized hosts and released into drinking water reservoirs through untreated sewage. Therefore, its proper identification is necessary when causing human gastrointestinal changes. The objective of this work is to analyze the presence of oocysts of Cryptosporidium spp. in recreational areas of the Guarapiranga Resevoir located in the state of São Paulo, Brazil. Water samples were collected and concentrated between July and November and analyzed through the technique of Ziehl Neelsen. Samples showed positive results for Cryptosporidium spp. and cyanobacteria, which are commonly found in the aquatic biota. The latter can be found in large quantities in eutrophic environments due to high amount of organic matter released in water bodies. Oocysts were observed in 60% of the analyzed samples and the distribution between positive and negative samples was not dependent on the month of sampling, but showed statistically significant differences according to the sampling site.

Developing and Testing a Real-Time Polymerase Chain Reaction to Identify and Quantify Bovine Respiratory Syncytial Viruses.

The bovine respiratory syncytial virus (BRSV) known as Bovine orthopneumovirus according to the international classification is one of the most important etiological agents of respiratory diseases in calves. At present, rapid and reliable methods to detect and measure the concentrations of this pathogen are needed. The objectives of the survey are developing the real-time polymerase chain reaction (PCR) to identify and quantify the BRSV RNA and, based on it, determining the number of the virus genomes in the respiratory tract of sick animals during the disease outbreaks. The nucleocapsid (N) protein gene of the virus served as the target for amplification. Messenger RNA (mRNA) of bovine GAPDH was used as a reference gene. A panel of positive control samples at known concentrations was used to estimate the virus and GAPDH numbers. The concentration of viral RNA extracted from the biomaterial samples was quantified relative to the bovine GAPDH mRNA level. The analytical sensitivity of PCR demonstrating high specificity and reproducibility was 1 × 103 genome equivalents per 1 cm3. All 273 samples of biological material taken from the animals with the respiratory diseases were analyzed. The virus genome was detected in 19.4% of samples. The viral RNA was more frequently detected in the lungs, which comprised 10.61% of positive samples. It was less frequently found in the mucous membranes of trachea and bronchi and the lymph nodes of the lungs, which comprised 0.73% of positive samples each. Concentrations of the virus in samples varied. The highest concentration was recorded in the lungs (1.3 ± 0.5—4.8 ± 0.47 log10 copies of BRSV/GAPDH RNA). The developed test kit may be used to quantify the concentration of the bovine respiratory syncytial virus in disease pathogenesis and to estimate the efficiency of vaccine or antivirus preparations for animals.

Results of scientific expedition to natural biotopes of the Republic of Tyva in 2019 with the purpose of infectious disease monitoring in wild bird populations

The results of the scientific expedition to Tere Khol and Uvs Nuur Lakes in the Republic of Tyva with the purpose of active monitoring of highly dangerous diseases in wild migratory waterfowl and epidemic analysis of these biotope water areas are presented in the paper. The Uvs Nuur Lake is a kind of an indicator for avian influenza introduction to the Russian Federation, because this is the resting and nesting area for many migratory wild birds during the period of mass migrations from Central and South-East Asian countries. In the process of active monitoring the complete autopsy of bird carcasses with description of organs and systems and sampling for laboratory diagnostics were performed. Droppings (pooled samples), parts of internal organs from dead and shot birds, blood (if possible) served as biological and pathological material for testing. While sampling, species were identified using an ornithological guide. The autopsy of dead waterfowl and birds shot for diagnostic purposes demonstrated a high worm burden of nematodes and cestodes. Two samples from European herring gulls were positive for avian influenza type A virus genome and subtype H13N6 was identified in one of them. Avian paramyxovirus serotype 1 (APMV-1), the agent of Newcastle disease, was found in one sample from gulls. The lakes of the Republic of Tyva are the most significant sites for sampling of biological material from wild birds, as the primary detection of highly pathogenic avian influenza virus in this territory is a serious signal of potential further virus spread and a precursor to a probable epizooty. Notwithstanding the absence of AIV very virulent isolate detections in wild bird populations the middle term prognosis for 2020 can be designated as cautious, as the avian influenza epidemic situation is deteriorating globally, especially in the European countries, and the threat of the virus introduction to the Russian territory with migratory birds still exists.

Банк биологического материала Северского биофизического научного центра

Purpose: Description of the collection, structure and development dynamics during the period from 2015 to 2019 of the biological material bank of the Seversk Biophysical Research Center, as well as research works carried out using biological material from its collection. &#x0D; Material and methods: The collection of bank of biological material includes the following types of biological material: venous blood, genomic DNA of white blood cells, cytogenetic suspensions of blood lymphocytes, surgical, biopsy and autopsy material. Biological material was obtained from people exposed to chronic radiation exposure: employees of the nuclear industry and the population permanently residing in the territory located in the area of the nuclear industry. &#x0D; Results: At present, the collection of the biological material bank contains more than 21,000 samples of biological material obtained from more than 8,000 donors. The collection of bank of biological material is represented by 4 main blocks: biological material of conditionally healthy employees of company of the nuclear industry (Siberian Chemical Combine); biological material of conditionally healthy people permanently residing in the territory located in the area of the nuclear industry enterprise (Seversk); biological material of patients (workers of the Siberian Chemical Combine and residents of Seversk) with malignant neoplasms and biological material of patients (workers of the Siberian Chemical Combine and residents of Seversk) with acute myocardial infarction. In the collection of bank of biological material, the proportion of samples of biological material of relatively healthy workers of the Siberian Chemical Combine is 33 %, residents of Seversk — 40.8 %, patients with malignant neoplasms — 22.2 %, patients with acute myocardial infarction — 4 %. &#x0D; Conclusion: Biological material bank Seversk Biophysical Research Center is unique collection of biological material samples of healthy people and patients with socially significant diseases. A feature of the biological material of the biological material bank is that it is obtained from people who have been exposed to chronic low-intensity radiation exposure and can be used to evaluate its radiogenic effects.

Exposure to mercury from dental amalgam: actual contribution for risk assessment.

Mercury dental amalgam restorations are an important source of chronic exposure to mercury in the whole population and special attention should be paid not only to occupational exposure to mercury during the preparation and administration of amalgam. The authors' report is an up-to-date contribution to the health risk assessment of mercury use in dentistry, namely occupational exposure to mercury in dentists working with dental amalgam and exposure to mercury in persons treated with amalgam dental restorations. Determination of total mercury in samples of biological material (urine, hair) was performed during 2017 and 2018 in 50 persons by the AAS method using the mercury vapour generation technique at 254.6 nm. Current dental exposures based on the most recent findings do not exceed acceptable risk levels and are below the biological limit of mercury in urine valid for occupationally exposed persons (100 μg.g-1 of creatinine), namely median value was 1.48 (min. < limit of detection (LOD), max. 17.14) μg.g-1 of creatinine (40 persons), total mercury content in hair of dental personnel expressed as median value was 0.340 (min. 0.060, max.1.628) μg.g-1. In controls (10 persons) was total mercury content in urine expressed as median value 0.36 (min. < LOD, max. 2.74) μg.g-1 of creatinine, in hair was median value 0.224 (min. 0.059, max. 0.453) μg.g-1. Authors support opinion that amalgam fillings in the oral cavity are a permanent source of mercury for the body itself.

Characteristic of urethral microbiota of men with idiopathic urethritis

Aim: to study the urethral microbiota composition in urogenital infections in men and to determine the sensitivity of isolated microorganisms to antibacterial drugs.Materials and Methods. 957 samples of biological material from the urethra of men were examined. The identification of the microorganisms was carried out according to the Bergey's Manual of Systematic Bacteriology. Sensitivity to antibiotics was determined by the disco-diffusion method, using standard disks of industrial production in accordance with the order of the Ministry of Health of Ukraine No. 167 of 05.04.2007. The mathematical processing of the results was performed, using the computer program Origin Pro 7.0.Results of the research. According to the study, the presence of conditionally pathogenic microorganisms was found in 716 people (74.82% of the total number of the surveyed contingent). Of the subjects studied, 1574 strains were isolated, of which the vast majority consisted of bacteria of the intestinal group (43.52%) and staphylococci (32.46%). The incidence of streptococci was 18.61%, corynebacteria - 3.81%, pseudomonads - 1.14%, candida - 0.44%. When studying the species composition of urethral microbiota, the presence of monoinfection was found in 267 people, in the other 449 persons different associations of representatives of conditionally pathogenic microbiota were detected. The analysis of the level of sensitivity of the studied strains to antibiotics showed that the isolated staphylococci showed the highest sensitivity to ampicillin / sulbactam (81.60% sensitive strains), most cephalosporins (79,26 ± 4,49%), carbapenems (93,35 ± 1,38 %), vancomycin (100%), rifampicin (92.56%), most fluoroquinolones (82.68 ± 3.31%) and linezolid (98.63). Whereas the highest number of susceptible strains of bacteria of the intestinal group was determined for carbapenems (89.64 ± 2.68%) and Co-trimoxazole (82.19%).Conclusions. Isolated bacterial strains of the intestinal group and staphylococci are most commonly found in association with other microorganisms. The analysis of the level of antibiotic sensitivity indicates that staphylococci showed the highest sensitivity to ampicillin / sulbactam, most cephalosporins, carbapenems, vancomycin, rifampicin, most aminoglycosides and linezolid. Whereas the highest number of susceptible strains of bacteria of the intestinal group was determined for carbapenems and Co-trimoxazole. Which makes the use of these antibiotics appropriate for the treatment of diseases of the men's genitourinary system

Does legislation affect elasmobranch conservation and research in Brazil? A case study from Paraná State

In Brazil, regulations for fishing for elasmobranchs are scarce, as well as insufficient monitoring and management. The first catch regulation for the group was published in 1998 (IBAMA No. 121/98). Despite the ban on finning, the legislation provides legal support for evisceration and decapitation prior to landing. Such practices have become particularly problematic for proper monitoring since decharacterization impairs the correct identification of species. In addition, processing is detrimental to research, as many studies rely on sampling of biological material (e.g., blood), posing an additional challenge for researchers using commercial fishing animals for scientific purposes. This study aimed to characterize the frequency of this practice in a representative artisanal fishing community in the fishing of elasmobranchs. The three years of monitoring show that landing patterns of gutted and beheaded animals are influenced by the species caught and the catch volumes of the day. In addition, endangered species protected by legislation were mostly landed already processed. We discuss the findings focusing on the need to revise the legal framework for the commercial landing of elasmobranchs and possible promising ways to address this challenge.

Diag nostics of gonorrhoea in Ukraine following World Health Organization recommendations

Objective — to evaluate the compliance of gonorrhoea diagnostics in regional skin and venereological dispensaries (RSVD) in Ukraine during 2013—2014 to World Health Organization (WHO) laboratory recommendations and identify areas for its improvement. Materials and methods. The study analyzed statistics data regarding the number of performed laboratory tests and their positive results for the detection of gonococci, the collection of biological samples and the use of laboratory methods for N. gonorrhoeae identification obtained from 14 RSVD of Ukraine by questionnaire. Results and discussion. Dispensary laboratories performed microscopy, culture and point-of-care-tests in 100, 85.7 and 7.1 % of patients, respectively. Gonococci were found in 2.3 % of the patients, equally by the results of microscopy (1740/76 348) and by culture (1322/56 273). Dispensary laboratories did not perform nucleic acid amplification tests. Sampling of biological material from three anatomical sites (urogenital, pharyngeal and rectal specimens) in heterosexual patients with symptoms of sexually transmitted infection was performed by 25 % of respondents, from two sites (urogenital and pharyngeal) — by 41.7 %, and from one site (urogenital sample) — by 33.3 % of dispensaries. External quality control of laboratory tests for the detection of N. gonorrhoeae was performed by 50 % of the respondents. Non-selective nutrient media for isolation of N. gonorrhoeae was used by 16.7 % of dispensaries, and prepared nutrient media in test tubes instead of the recommended Petri dishes, were used in 58.3 % of dispensaries. The vast majority of respondents (78.6 %) had an experience of determining gonococcal sensitivity to antibiotics. Two institutions (14.3 %) participated in an international pilot project to determine the antibiotic resistance of gonococci. Conclusions. Diagnostics of gonococcal infection in Ukraine is not optimal and needs improvement by using quality-tested bacteriological cultures and point-of-care-tests, implementation of nucleic acid amplification tests, monitoring of gonococcal sensitivity to antibiotics, collection of extragenital biological samples. Adherence to evidence-based international guidelines and adapted national guidelines for the diagnostics of gonorrhoea is an essential factor of the controlling of gonococcal infection.

Specific composition of indigenous microflora (Lactobacillus spp., Bifidobacterium spp., Lactococcus spp.) in farm animals

To maintain a stable composition of the gastrointestinal tract microflora in farm animals it is necessary to use probiotic agents to ensure the full functioning of the digestive, hormonal, and immune systems of the body. Most modern probiotics include lactic acid bacteria and bifidobacteria, which are the most physiologically valuable components of a healthy organism’s an indigenous microflora. The aim of this study was to provide indication and identification from the milk of healthy cows and gastric tract of healthy pigs and calves of the genus bacteria Lactobacillus, Bifidobacterium, and Lactococcus. The objects of research were cultures of microorganisms isolated from cows milk (82), the gastrointestinal tract of cattle (317), and piglets of different age groups (114). Bacteriological studies were carried out on the basis of the veterinary sanitation and parasitology laboratory of the National Scientific Center "Institute of Experimental and Clinical Veterinary Medicine" (Kharkiv) in accordance with current regulatory documents. According to the research of the gastrointestinal tract of clinically healthy calves and piglets isolated and typified to 317 and 114 cultures of microorganisms, the species composition of the microflora (82 bacterial cultures) of the cisternous and parenchymatous milk of clinically healthy cows was determined. A total of 513 isolates of microorganisms were isolated, including: Enterobacter spp. –2 (0,39%), Staphylococcus spp. – 7 (1,37%), Bacillus spp. – 11 (2,14%), Enterococcus spp. – 33 (6.43%), Lactococcus spp. – 75(14,62%), Bifidоbacterium spp. – 170 (33,14%), and Lactobacillus spp. – 215 (41,91%). In the study of the biological properties of isolated microorganisms Lactobacillus spp. (215) established their species identity: L. brevis – 7 (3.26%), L. delbrueckii – 9 (4,19%), L. acidophilus – 21 (9,77%), L. fermentum – 23 (10,69%), L. casei – 57 (26,51%), and L. plantarum – 98 (45.58%). Cultures of Bifidobacterium spp. (170) belong to B. suis – 2 (1,18%), B. breve – 7 (4,12%), B. lactentis – 15 (8,82%), B. bifidum – 21 (12,35%), B. longum – 22 (12,94%), B. infantis – 25 (14,71%), and B. adolescentis – 78 (45,88%). From samples of biological material of farm animals, 75 cultures of the genus Lactococcus spp. were isolated (75) of which Lactococcus lactis is representative. Isolated bacteria Lactobacillus spp., Bifidobacterium spp. and Lactococcus spp. promising when creating innovative probiotic products for farm animals.

DNA barcoding in some Belarusian insects

Due to a number of factors in the anthropogenic load on ecosystems, environmental changes, and competition between native and invasive species, the loss of species diversity has currently risen to an alarming scale. One of the prospective approaches to remedy the current state and halt the loss is the international project “Barcode of Life”. The report describes the results of molecular genetic research on the insect representatives of such orders as Coleoptera (beetles), Lepidoptera (butterflies), and Trichoptera (caddisflies) using the cytochrome c oxidase subunit 1 of the mitochondrial region (COI). We proposed a technique for the sampling of biological material to isolate DNA from the hind legs of individuals. The technique prevents removal of the individuals of the most valuable species collected on the territory of the Republic of Belarus from their habitats. The DNA sample collection of order representatives was investigated. For the subfamily Cetoniinae (flower chafer), significant differences in haplotypes among the representatives of the Belarusian and European parts of their areal were found. For Trichoptera, it was shown that the COI gene has high variability to differentiate species. It was also revealed that some species of caddisflies, which Belarusian researchers believe to be synonymous ones, have sequences with big differences according to the BoldSystem database. This fact should be explored in the future. So, our analysis allows considering the COI gene as a satisfactory marker for the species identification in the Belarusian insects’ taxa studied.

ПОВЕДЕНИЕ ЭЛЕМЕНТОВ ПРИ ВЫСОКОТЕМПЕРАТУРНОМ ОЗОЛЕНИИ КОСТНОЙ ТКАНИ

The number of studies of ashed biosubstrates in terms of elemental composition is sharply increasing due to the growing interest of living matter. Thus, it is extremely important to know, is the composition of the ashed material reflects the composition of the original object and how elements behave during the process of combustion. To answer these questions, an experiment on the samples of biological material was made. The purpose of the study is to analyze the behavior of elements in the process of high-temperature combustion of bone tissue. During the experiment, the following methods of analysis were used: instrumental neutron activation and mass spectrometry with inductively coupled plasma. The study shows that most of the elements are concentrated in bone ash during combustion (Li, Na, Mg, P, Ca, Ti, Cu, Zn, Ga, U, Al, Sc, Cr, Mn, Fe, As, Y,, Zr , Nb, Mo, Cd, Sb, La, Ce, Pr, Sm, Gd, Au, W, Bi), while volatilization is observed only for Br, Hg and Sn. The exclusive role of the chemichal compound of the element is shown for the analysis of its behavior. The experiment is aslo shows the role of combustion temperature to predict the behavior of element: it was found that lower temperature of ashing (500 °C) leads to more intensive volatilization of elements. In addition, an algorithm is propesed to convert the content of the element in ash of the bone to dry matter. At the same time, the dependence of the content of elements in the ash on their content in dry tissue were established using mathematical functions with high accuracy of approximation (for Fe, Mg, Zn, As, Pr, Ce, W, Zr, Cd, Cr, Sb, Mn, Ga, Nb, Pb, and U).

Geographic information mapping methods as a basis for systematization of environmental monitoring data on the Angara River system

This work proposes a model for the application of cartographic and geographic information mapping methods to systematize environmental monitoring data. In particular, based on the results of sampling and study of biological material (roach, Rutilus rutilus (Cyprinidae, Cypriniformes)) from the Angara River system, we obtained an array of information this could be processed and organized on its basis by means of a cartographic presentation of research materials on the state of the environment of water bodies in the Irkutsk region.

Analysis of the linkage between viral, bacterial, and protozoal infections and the level of reproductive losses in dairy herds in the Ural region

The analysis of the linkage between infection of dairy herds in the Ural region with BVDV, BHV-1, C. abortus and N. caninum with indices of reproductive function and reproductive losses was performed. The results of studies of 1453 samples of biological material by Elisa, PCR and Real-Time PCR were taken into account. The level of BVDV infection in animals of various physiological groups in the examined farms ranged from 15.6 to 66.7%, and the seroprevalence of the BHV-1 field strain was 31.8%. The amount of seroprevalent to C. abortus animals in dairy herds was 55.1 %, to N. caninum-43.3%. The lowest rates of fertilization were found in farms with simultaneous circulation of chlamydia and neosporosis pathogens and in mono-chlamydia infection. In these same herds, there was a longer post-partum interval. The highest mean rate of abortions in cows and heifers was in farms with N. caninum circulation, as well as in mixed infections, with increased action of infectious factors. In the analysis of viral infections, it was established that the highest level of perinatal losses was seen in the presence of BVDV.

Instrument to streamline the process of sending biological samples to an epidemiological surveillance center, from a Reference Unit for infectious and parasitic diseases.

Introduction: The sending of biological materials for specific exams to the Public Health Laboratory of Amazonas (LACEN-AM) depends on documents containing essential information to guide the performance of the tests, which need to go through a quality process and in most cases the lack of data compromises the streamflow to LACEN-AM. Objective: Create a checklist for the process of sending samples to the Public Health Laboratory (LACEN-AM). Methods: The Health Surveillance Guide (GVS), Integrated Manual for Epidemiological Surveillance of botulism, technical notes, reference technicians for information on registration and tests performed at LACEN-AM were consulted and the sequence of activities was also observed. carried out in the notification epidemiology sector, the orientation of the collection of biological material until the sending of the samples. Results: Between 2017 and 2019, 6 lists were sent to send samples to LACEN-AM, one list for each disease (Paracoccidioidomycosis, Hydatidosis, Severe Acute Respiratory Syndrome, Acute Flaccid Paralysis, Human Rabies and Botulism). Conclusion: It is necessary to simplify the procedures related to the notification activities for sending biological material samples, improving operationalization, especially for paracocociodiomycosis, hydatidosis and SRAG, diseases that are not included in the GVS, making it difficult to understand the flow of sending biological materials when requested or, in the cases of (PFA), human rabies and botulism which, because they are in the process of monitoring, have their flows difficult to discern. Therefore, disseminating these procedures by means of an instrument can contribute to the improvement and understanding of the technicians who work in epidemiological surveillance, and will certainly speed up their actions, ensuring speed in sending the samples.

Comparative Characteristics of the Culture-enzymatic and Molecular-genetic Methods for Indicating Mollicutes – Mycoplasma hominis and Ureaplasma spp., Isolated in Patients with Acute Uncomplicated Pyelonephritis

The objective: is to analyze the frequency of detection of M.hominis and Ureaplasma spp. in the biological material of patients with acute uncomplicated pyelonephritis (AUP) by cultural-enzymatic and molecular-genetic methods and to prove their involvement in the development of the inflammatory process of the urinary and genital tracts.Materials and methods. The study involved 224 women of reproductive age, patients with AUP and concomitant chronic diseases of the pelvic organs. We examined urine samples, scrapings of the urethral mucosa and cervical canal using cultural-enzymatic (CEM) and polymerase chain reaction (PCR). Іn the serum of 217 patients determined the level of specific IgG-antibodies to mollicutes by enzymelinked immunosorbent assay.Results. The introduction of the PCR method for the detection of mollicutes in the urinary and genital tracts in patients with AUP revealed possible to increase the efficiency of their indication, primarily when M.hominis is detected. The frequency of Ureaplasma spp. detection by both methods was significantly higher than M.hominis in samples of biological material of different localization in the examined patients. A high frequency of coincidence in the detection frequency of M.hominis and Ureaplasma spp. was noted using two methods (PCR and CЕM): in the urine – 85.9% and 87.5%, in scrapings of the mucous membrane of the urethra – 81.3% and 93.3% and the cervical canal – 78.0% and 88.8%. The coincidence of diagnostic levels of antibodies to Ureaplasma spp. in the presence of this pathogen in the biological material studied in 65 (76.5%) of 85 patients were registered, and M.hominis – in 34 (41.0%) of 83 patients.Conclusion. The complex use of various methods of laboratory diagnostics allows obtaining accurate data on the presence of mycoplasma infection in patients with AUP and concomitant chronic diseases of the pelvic organs. The presence of diagnostically significant titers of IgG-antibodies to M.hominis and Ureaplasma spp. proves their influence on the course of the inflammatory process of the urinary and genital tracts of women with AUP.

Granulation tissue morphology of the laparotomic wound with different types of sutures

The aim — to study experimentally the granulation tissue morphological features of the laparotomy wound after suturing it by simple knots and continuous wrapping sutures.Materials and methods. The experiment was performed on 108 rats undergoing a median laparotomy. The first group of animals (54 rats) edges of the musculoaponeurotic layer of the anterior abdominal wall were connected by simple sutures and in the second group (54 rats) by continuous wrapping. A sampling of biological material was performed on the 1st, 7th and 14th days after surgery, by excision of the musculoaponeurotic layer of the anterior abdominal wall along with the skin.Results and discussion. On the 1st day of the study, in the first group, lymphoid cells and neutrophils statistically significantly prevailed in the granulation tissue, the difference between their number was statistically insignificant.The second group of animals is likely to have a higher percentage of lymphoid cells only. At the 7th and 14th days of observation, the percentage of fibroblasts in the granulation tissue of animals of both groups probably prevails. On the 1st day after surgery, fibrin is observed in both experimental groups of animals, in the form of a grid, and in the second experimental group, there are still hemorrhages. The morphology of the laparotomy wound granulation tissue for the 14th day of observation does not differ when applying a simple knot and continuous suture, except that the latter is likely to be dominated by the specific volume of collagen fibers.Conclusions. The use of continuous suture, unlike simple knots, on the 14th day after suturing of the laparotomy wound lead to a probable increase in the percentage of fibroblasts, a decrease in macrophages, as well as a predominance of the specific volume of collagen fibers, indicating a relatively rapid maturation of the tissue.

Statistical modeling of STR capillary electrophoresis signal

BackgroundIn order to isolate an individual’s genotype from a sample of biological material, most laboratories use PCR and Capillary Electrophoresis (CE) to construct a genetic profile based on polymorphic loci known as Short Tandem Repeats (STRs). The resulting profile consists of CE signal which contains information about the length and number of STR units amplified. For samples collected from the environment, interpretation of the signal can be challenging given that information regarding the quality and quantity of the DNA is often limited. The signal can be further compounded by the presence of noise and PCR artifacts such as stutter which can mask or mimic biological alleles. Because manual interpretation methods cannot comprehensively account for such nuances, it would be valuable to develop a signal model that can effectively characterize the various components of STR signal independent of a priori knowledge of the quantity or quality of DNA.ResultsFirst, we seek to mathematically characterize the quality of the profile by measuring changes in the signal with respect to amplicon size. Next, we examine the noise, allele, and stutter components of the signal and develop distinct models for each. Using cross-validation and model selection, we identify a model that can be effectively utilized for downstream interpretation. Finally, we show an implementation of the model in NOCIt, a software system that calculates the a posteriori probability distribution on the number of contributors.ConclusionThe model was selected using a large, diverse set of DNA samples obtained from 144 different laboratory conditions; with DNA amounts ranging from a single copy of DNA to hundreds of copies, and the quality of the profiles ranging from pristine to highly degraded. Implemented in NOCIt, the model enables a probabilisitc approach to estimating the number of contributors to complex, environmental samples.