Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain.

BACKGROUND: Bovine genital campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis (Cfv) including its biovar intermedius (Cfvi). This sexually transmitted disease induces early reproductive failure causing considerable economic losses in the cattle industry. Using a collection of well-characterized isolates (n = 13), C. fetus field isolates (n = 64) and saprophytic isolates resembling Campylobacter (n = 75) obtained from smegma samples of breeding bulls, this study evaluated the concordance of the most used phenotypic (H2S production in cysteine medium and 1% glycine tolerance) and molecular (PCR) methods for the diagnosis of BGC and assessed possible cross-reactions in the molecular diagnostic methods. RESULTS: Characterization at the subspecies level (fetus vs. venerealis) of C. fetus isolated from bull preputial samples using phenotypic and molecular (PCR targeting nahE and ISCfe1) methods showed moderate concordance (κ = 0.462; CI: 0.256-0.669). No cross-reactions were observed with other saprophytic microaerophilic species or with other Campylobacter species that can be present in preputial samples. Whole genome sequencing (WGS) of discrepant isolates showed 100% agreement with PCR identification. For the differentiation of Cfv biovars, comparison of the H2S test (at 72 h and 5 days of incubation) and a PCR targeting the L-cysteine transporter genes showed higher concordance when H2S production was assessed after 5 days (72 h; κ = 0.553, 0.329-0.778 CI vs. 5 days; κ = 0.881, 0.631-1 CI), evidencing the efficacy of a longer incubation time. CONCLUSIONS: This study confirmed the limitations of biochemical tests to correctly identify C. fetus subspecies and biovars. However, in the case of biovars, when extended incubation times for the H2S test (5 days) were used, phenotypic identification results were significantly improved, although PCR-based methods produced more accurate results. Perfect agreement of WGS with the PCR results and absence of cross-reactions with non-C. fetus saprophytic bacteria from the smegma demonstrated the usefulness of these methods. Nevertheless, the identification of new C. fetus subspecies-specific genes would help to improve BGC diagnosis.

Metabolic causes of pediatric developmental & epileptic encephalopathies (DEE)- genetic variant analysis in a south Indian cohort.

PURPOSE: Drug-resistant epilepsy is seen in patients with inborn errors of metabolism and metabolic dysfunction in neurons is crucial to brain disorders associated with psychomotor impairment. Diagnostic rates of metabolic causes of developmental and epileptic encephalopathy (DEE) using next generation sequencing have been rarely studied in literature. METHODS: A prospective hospital study was carried out in 384 children with DEE, who underwent genetic testing. Metabolic disorders were evaluated with biochemical blood/urine assays and when required CSF estimations performed. RESULTS: A total of 154 pathogenic/likely pathogenic variants in 384 children were identified. Out of 384 children, 89 were clinically suspected to have probable or possible metabolic disorders. Pathogenic/likely pathogenic variants in metabolic genes were identified in 39 out of 89 (43.8 %) and promising VUS in 28 (31.4 %). These included variants for progressive myoclonus epilepsies (21; 53.8 %), DEE with focal/multifocal seizures (8; 20.5 %), generalized epilepsy (5;12.8 %), early myoclonic encephalopathy (2; 5.1 %), LGS (1; 2.6 %) and West syndrome (2; 5.1 %). CONCLUSION: Our cohort demonstrates for the first time from the Indian subcontinent that identification of metabolic variants can guide investigations and has therapeutic implications in patients with variable DEE phenotypes. A high utility is noted with regard to diagnosis and prognostication, given the low yield of available biochemical tests, indicating cost-effectiveness of this approach.

The predictive value of liver tests for the presence of liver metastases.

Aim: To analyze the predictive value of biochemical liver tests in patients with malignant melanoma, breast, colorectal or lung cancers at the time of diagnosis of liver metastases. Methods: A retrospective review of patients with the above-mentioned solid tumors at MedStar Georgetown University Hospital from 2016-2020. Results: The highest optimal cutoff according to sensitivity and specificity for the presence of liver metastases was for AST ≥1.5 × ULN for melanoma, lung, and breast cancers and ≥2 × ULN for colorectal cancer, ALT ≥1.25 × ULN for melanoma, breast and colorectal cancers and ≥1.5 × ULN for lung cancer, and ALP ≥1.5 × ULN for melanoma, breast and colorectal cancers. Conclusion: Using thresholds of liver enzymes above the ULN may improve the diagnostic accuracy for the presence of liver metastases.

Isolation and Molecular Identification of Salmonella pullorum from Broiler Chicken in Iraqi Fields.

Pullorum disease (PD) is one of the most common diseases in the world, with devastating consequences. In the chicken sector, there have been financial losses. It is brought on by Salmonella enteric subspecies serovar Gallinarum biovar pullorum; definitive detection requires culture followed by biochemistry analysis and serotyping. This study aimed to verify the presence of bacteria by culture, biochemical characterization, PCR assay, and sequencing. One hundred samples were collected from 12 broiler chicken flocks of different ages for 8districts of Baghdad province, including cloacal swabs (65), visceral organs (15), and dropping (20). Salmonella colonies were identified by selective culture broth and agar with biochemical description for 75% of the total samples, with a higher incidence in visceral organs than dropping and cloacal swabs. ،The Sequencing and phylogenetic tree analysis of 16S rRNA gene for representative Salmonella isolates. The presence of Salmonella pullorum isolates in global genetic strains; was revealed a matching NCBI isolates similarity of 99.02% with (MF445124.1) and 98% with (MH352164.1), respectively. In the current state of molecular and genetic research, phlyogentic research announced the real presence of Salmonella pullorum in Baghdadprovince's broiler chicken, also showing the phylogentic characteristics and links to some global isolates. The detection of Salmonella pullorum in broiler flocks of the current study extent of health risks to other uninfected birds present in the free range.

Coffee intake one hour prior to phlebotomy produces no clinically significant changes in routine biochemical test results.

Introduction: Although current guidelines recommend not drinking coffee prior to phlebotomy, our hypothesis is that drinking coffee does not affect the clinical interpretation of biochemical and haematological test results. Materials and methods: Twenty-seven volunteers were studied in basal state (T0) and 1h after (T1) drinking coffee. Routine haematological (Sysmex-XN1000 analyser) and biochemistry parameters (Vitros 4600 analyser) were studied. Results were compared using the Wilcoxon test (P < 0.05). A clinical change was considered when mean percent difference (MD%) was higher than the reference change value (RCV). Results: Coffee intake produced statistically, but not clinically, significant: i) increases in haemoglobin (P = 0.009), mean cell haemoglobin concentration (P = 0.044), neutrophils (P = 0.001), albumin (P = 0.001), total protein (P = 0.000), cholesterol (P = 0.025), high density lipoprotein cholesterol (P = 0.007), uric acid (P = 0.011), calcium (P = 0.001), potassium (P = 0.010), aspartate aminotransferase (P = 0.001), amylase (P = 0.026), and lactate dehydrogenase (P = 0.001), and ii) decreases in mean cell volume (P = 0.002), red cell distribution width (P = 0.001), eosinophils (P = 0.002), and lymphocytes (P = 0.001), creatinine (P = 0.001), total bilirubin (P = 0.012), phosphorus (P = 0.001), magnesium (P = 0.007), and chloride (P = 0.001). Conclusion: Drinking a cup of coffee 1 hour prior to phlebotomy produces no clinically significant changes in routine biochemical and haematological test results.

Turmeric-Associated Liver Injury: A Rare Case of Drug-Induced Liver Injury.

Turmeric is popularly used as a naturopathic supplement associated with myriad benefits and has long been generally regarded as safe. However, increasing reports of turmeric-associated liver injury have emerged over recent years. This case presents a female patient without significant past medical history who presents with signs and symptoms of acute hepatitis after consuming a turmeric-containing tea. Her case adds to a growing body of evidence that dosage safety, manufacturing, and pharmacologic delivery practices for turmeric supplements should be investigated.

A Previously Undiagnosed Case of Alkaptonuria in an 80-Year-Old Patient: A Case Report.

Alkaptonuria is a rare genetic metabolic disorder of autosomal recessive inheritance characterised by the accumulation of homogentisic acid in the body. It is diagnosed upon identification of characteristic symptoms, using various biochemical investigations, radiographic pictures, and a variety of specialised tests. Here we are discussing the case of an 80-year-old female patient with incidental findings of alkaptonuria. It is crucial to understand the fundamental diagnostic investigations that can be used in low-income nations or facilities where investigations like genetic testing, gas chromatography, and mass spectrometry are not readily available for the diagnosis of alkaptonuria.

Biochemical indexes and gut microbiota testing as diagnostic methods for Penaeus monodon health and physiological changes during AHPND infection with food safety concerns.

Severe shrimp disease outbreaks have a destructive impact on shrimp aquaculture and its associated downstream food processing industries. Thus, it is essential to develop proper methods for shrimp disease control, which emphasizes the importance of food safety. In this study, we performed biochemical tests and gut microbiome analysis using uninfected control and Vp AHPND-infected Penaeus monodon samples. Biochemical tests were performed to assess the phenoloxidase (PO) activity, respiratory Burst (RB) activity, nitrite concentration, superoxide dismutase (SOD) activity, total hemocyte count (THC), and total protein concentrations. Overall, upregulations were detected in these biochemical tests, which showed the activation of the immune response in P. monodon during acute hepatopancreatic necrosis disease (AHPND) infection, especially at 6 hpi and 12 hpi. Besides that, shrimp gut samples were collected and pooled (n = 3), followed by DNA extraction, PCR amplification targeting the V3/V4 16S ribosomal RNA (rRNA) region, next-generation sequencing (NGS), and bioinformatics analysis. Proteobacteria was the most abundant phylum in both samples. The Rhodobacteraceae family and Maritimibacter genus were proposed to be vital forshrimp health maintenance. Vp AHPND bacterial colonization and secondary Vibrio infections were postulated to have occurred based on the higher abundances of Vibrionaceae family and Vibrio genus in the Vp AHPND-infected sample. Firmicutes phylum together with Photobacterium and Aliiroseovarius genera were inferred to be pathogenic or related factors of AHPND infections. In conclusion, physiology (immune response activation) and gut microbiome changes of disease tolerant P. monodon during AHPND infection were identified. Both biochemical tests and 16S rRNA analysis are proposed as a combined strategy for shrimp health diagnosis for ensuring shrimp health maintenance, disease control, and food safety.

Co-Inheritance of Variation in All-Cause Mortality and Biochemical Risk Factors.

Biomarkers may be useful endophenotypes for genetic studies if they share genetic sources of variation with the outcome, for example, with all-cause mortality. Australian adult study participants who had reported their parental survival information were included in the study: 14,169 participants had polygenic risk scores (PRS) from genotyping and up to 13,365 had biomarker results. We assessed associations between participants' biomarker results and parental survival, and between biomarker results and eight parental survival PRS at varying p-value cut-offs. Survival in parents was associated with participants' serum bilirubin, C-reactive protein, HDL cholesterol, triglycerides and uric acid, and with LDL cholesterol for participants' fathers but not for their mothers. PRS for all-cause mortality were associated with liver function tests (alkaline phosphatase, butyrylcholinesterase, gamma-glutamyl transferase), metabolic tests (LDL and HDL cholesterol, triglycerides, uric acid), and acute-phase reactants (C-reactive protein, globulins). Association between offspring biomarker results and parental survival demonstrates the existence of familial effects common to both, while associations between biomarker results and PRS for mortality favor at least a partial genetic cause of this covariation. Identification of genetic loci affecting mortality-associated biomarkers offers a route to the identification of additional loci affecting mortality.

A dPCR-NIPT assay for detections of trisomies 21, 18 and 13 in a single-tube reaction-could it replace serum biochemical tests as a primary maternal plasma screening tool?

BACKGROUND: The next generation sequencing (NGS) based non-invasive prenatal test (NIPT) has outplayed the traditional serum biochemical tests (SBT) in screen of fetal aneuploidies with a high sensitivity and specificity. However, it has not been widely used as a primary screen tool due to its high cost and the cheaper SBT is still the choice for primary screen even with well-known shortages in sensitivity and specificity. Here, we report a multiplex droplet digital PCR NIPT (dPCR-NIPT) assay that can detect trisomies 21, 18 and 13 (T21, T18 and T13) in a single tube reaction with a better sensitivity and specificity than the SBT and a much cheaper price than the NGS-NIPT. METHODS: In this study, the dPCR-NIPT assay's non-clinical characteristics were evaluated to verify the cell free fetal DNA (cffDNA) fraction enrichment efficiencies, the target cell free DNA (cfDNA) concentration enrichment, the analytical sensitivity, and the sample quality control on the minimum concentration of cfDNA required for the assay. We validated the clinical performance for this assay by blindly testing 283 clinical maternal plasma samples, including 36 trisomic positive samples, from high risk pregnancies to access its sensitivity and specificity. The cost effectiveness of using the dPCR-NIPT assay as the primary screen tool was also analyzed and compared to that of the existing contingent strategy (CS) using the SBT as the primary screen tool and the strategy of NGS-NIPT as the first-tier screen tool in a simulating situation. RESULTS: For the non-clinical characteristics, the sample processing reagents could enrich the cffDNA fraction by around 2 folds, and the analytical sensitivity showed that the assay was able to detect trisomies at a cffDNA fraction as low as 5% and the extracted cfDNA concentration as low as 0.2 ng/µL. By testing the 283 clinical samples, the dPCR-NIPT assay demonstrated a detection sensitivity of 100% and a specificity of 95.12%. Compared to the existing CS and the NGS-NIPT as the first-tier screen strategy, dPCR-NIPT assay used as a primary screen tool followed by the NGS-NIPT rescreen is the most economical approach to screen pregnant women for fetal aneuploidies without sacrificing the positive detection rate. CONCLUSION: This is the first report on a dPCR-NIPT assay, consisting of all the necessary reagents from sample processing to multiplex dPCR amplification, can detect T21, T18 and T13 in a single tube reaction. The study results reveal that this assay has a sensitivity and specificity superior to the SBT and a cost much lower than the NGS-NIPT. Thus, from both the test performance and the economic benefit points of views, using the dPCR-NIPT assay to replace the SBT as a primary screen tool followed by the NGS-NIPT rescreen would be a better approach than the existing CS for detection of fetal aneuploidies in maternal plasma.

Verificação dos parâmetros bioquímicos, lipídico e glicêmico em pacientes submetidos à hemodiálise em um hospital-escola do noroeste paulista / Verification of biochemical, lipid and glycemic parameters in patients undergoing hemodialysis in a teaching hospital in the noroeste paulista / Verificación de parámetros bioquímicos, lipídicos y glucémicos en pacientes sometidos a hemodiálisis en un hospital-escuela de la región noroeste de São Paulo

Introdução: A doença renal crônica consiste na alteração da função renal de forma lenta, irreversível e progressiva, que provoca danos na estrutura dos néfrons, levando à diminuição da capacidade de depuração do sangue. Isso resulta em desequilíbrio hidroeletrolítico e distúrbios bioquímicos, metabólicos, hormonais e acúmulo de substâncias tóxicas. Para esses casos é indicada a hemodiálise, uma terapia renal substitutiva, capaz de remover os metabólitos tóxicos do sangue. No entanto, ao longo do tempo, podem ocorrer alterações significativas nos exames bioquímicos dos pacientes. Objetivo: O objetivo deste trabalho foi coletar e analisar os dados dos prontuários dos pacientes submetidos à hemodiálise, referentes aos exames bioquímicos, lipídicos e glicêmicos e correlacionar as possíveis alterações bioquímicas com os valores de referência nos pacientes em hemodiálise. Material e Método: Trata-se de um estudo quantitativo, realizado em um hospital escola do interior paulista. No estudo, foram analisados 54 prontuários (n = 54) no período de fevereiro de 2020 a fevereiro de 2021, com faixa etária acima de 18 anos. Os dados colhidos nos prontuários foram referentes ao cálcio, fósforo, potássio, creatinina, ureia, glicose e lipídio. Resultados: Os resultados indicaram que todos (100%) os pacientes submetidos à hemodiálise apresentam alterações bioquímicas em todos os parâmetros, principalmente na creatinina e ureia pré-diálise, tanto no sexo masculino como no feminino. Conclusão: Analisando os resultados, ficou evidente o perfil do paciente hemodialítico e, também, a incidência de alteração para cada tipo de exame realizado no decorrer da hemodiálise, dada a sua importância na qualidade de vida do paciente.(AU)

Introduction: Chronic kidney disease consists of a slow, irreversible, and progressive alteration of the renal function, which causes damage to the structure of the nephrons, leading to a decrease in the blood clearance capacity. This results in hydroelectrolytic imbalance and biochemical, metabolic, and hormonal disturbances, and accumulation of toxic substances. Hemodialysis, a renal replacement therapy, is indicated for these cases and is able to remove toxic metabolites from the blood. However, over time, significant changes may occur in the biochemical tests of patients. Objective: The objective of this work was to collect and analyze data from the medical records of patients undergoing hemodialysis, regarding biochemical, lipid and glycemic tests and correlate the possible biochemical alterations with the reference values in hemodialysis patients. Material and Method: This is a quantitative study, carried out in a teaching hospital in the interior of São Paulo. In the study, 54 medical records (n = 54) were analyzed from February 2020 to February 2021, aged over 18 years. The data collected from the medical records were related to calcium, phosphorus, potassium, creatinine, urea, glucose and lipid. Results: The results indicated that all (100%) patients undergoing hemodialysis present biochemical alterations in all parameters, mainly in pre-dialysis creatinine and urea, both in males and females. Conclusion: Analyzing the results, the profile of the hemodialysis patient became evident, as well as the incidence of alteration for each type of exam performed during hemodialysis, given its importance in the quality of life of the patient.(AU)

Introducción: La enfermedad renal crónica consiste en una alteración lenta, irreversible y progresiva de la función renal, que provoca daños en la estructura de las nefronas, lo que conlleva una disminución de la capacidad de depuración de la sangre. Esto provoca un desequilibrio hidroelectrolítico y alteraciones bioquímicas, metabólicas y hormonales, así como la acumulación de sustancias tóxicas. Para estos casos, está indicada la hemodiálisis, una terapia de sustitución renal, para eliminar los metabolitos tóxicos de la sangre. Sin embargo, con el tiempo, pueden producirse alteraciones significativas en los exámenes bioquímicos de los pacientes. Objetivo: El objetivo de este trabajo fue recoger y analizar los datos de las historias clínicas de los pacientes sometidos a hemodiálisis, referentes a las pruebas bioquímicas, lipídicas y glucémicas y correlacionar las posibles alteraciones bioquímicas con los valores de referencia en los pacientes en hemodiálisis. Material and Método: Se trata de un estudio cuantitativo, realizado en un hospital...(AU)

Influence of the Oral Administration of Fish Oil on Hematological and Biochemical Performance in Local Male Rabbits.

In some developing countries in Africa, Asia, and Latin America, rapid population growth is a complicated problem, and thus, it is necessary to raise the efficiency of foodstuff production to preserve human life. Therefore, this study was designed to investigate the effect of fish oil supplementation on male rabbits' productivity, as well as their biochemical and physiological characteristics. This experiment was conducted in the Animal House of the College of Veterinary Medicine, University of Baghdad (Baghdad, Iraq) for eight weeks. Twenty-four local male rabbits were randomly divided into three groups (each containing eight rabbits) as follows: the control group that was fed with a basal diet only, the first treated group in which each animal received fish oil orally at a dose of 0.75 ml every day, and another treated group in which each animal received fish oil orally at a dose of 1.5 ml every day. Both treated groups received the treatment for 60 days. Blood samples were collected from cardiac veins by cardiac puncture at the beginning of the experiment (Zero time). On day 60 of the experiment, samples were collected again to analyze potential changes in blood characteristics, including white blood cells count, hemoglobin, and red blood cells count. These blood samples recorded a significant decrease in cholesterol in the two treatment groups, compared to the control. In conclusion, the current findings recommend daily oral administration of fish oil at a dose of 0.75 mg/ml or 1.5 mg/ml to rabbits for 60 days before conception to improve performance trials, as well as biochemical and hematological results. These alterations, however, have a small impact on these features, compared to higher doses administered to rabbits.

Detection of Salmonella spp. by Traditional and PCR Assays in Raw Milk, Maysan, Iraq.

Salmonella spp are characterized as rod- shaped, motile, gram- negative bacteria which has the ability to infect animals and human. Salmonella spp occasionally causes sickness while in most cases not lead to severe symptoms. Analyzing milk for Salmonella spp. is not routine but traditional culture methods are used to evaluate the health condition of the dairy products. However, the antibody-based and nucleic-acid- based methods are practical for identifying Salmonella spp. Therefore, this research was designed to evaluate the use of traditional culture methods and PCR in detection of the presence of Salmonella spp. in raw milk samples in, Maysan Iraq. A total number of 130 raw milk samples collected from Maysan Iraq. All the samples were analyzed for the presence of Salmonella spp. using traditional culture method and polymerase chain reaction (PCR). The culture method used in this experiment were done by using pre-enrichment, enrichment, selective plating and biochemical tests. The results of this traditional technique were compared with the results obtained from PCR method. The PCR was performed using a 284bp sequence of the invA gene. The results showed that 8 (7.07%) of samples were identified as salmonella positive using traditional culture technique but 14 (12.3%) samples were detected as salmonella positive by PCR method. The results of the current research revealed that the traditional culture based methods are generally time costuming and labor intensive but the development of new rapid methods including DNA based methods such as PCR are more sensitive and have dramatically decreased the time necessary for the detection of bacteria.

Evaluation of presumptive identification of Enterobacterales using CHROMagar Orientation medium and rapid biochemical tests.

BACKGROUND: The use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is gradually spreading among large-scale laboratories; however, this method is impractical for small-scale laboratories. In laboratories without access to these rapid identification methods, problems related to them remain unsolved. In this study, we aimed to develop a rapid and inexpensive method to presumptively identify Enterobacterales using CHROMagar Orientation medium. METHODS: The algorithm for presumptive identification of Enterobacteriaceae using CHROMagar Orientation medium was based on our previous studies. Modified property tests for indole, lysine decarboxylase, ornithine decarboxylase, and hydrogen sulfide were performed to evaluate the differentiation of the bacterial species. RESULTS: Using the type strains and clinical isolates, it was possible to conduct the property tests at a low cost, within 4 hours. The spot indole test was performed without any nonspecific reactions for the bacteria forming colored colonies. The presumptive identification of bacteria was thereby possible within 24 hours after specimen submission. CONCLUSION: All these results suggest that the rapid presumptive identification of Enterobacterales is possible with this new identification method using CHROMagar Orientation medium. This is therefore a prompt and economical method that can be used in routine laboratory work.

Morphological, biochemical and molecular identification of petroleum hydrocarbons biodegradation bacteria isolated from oil polluted soil in Dhahran, Saud Arabia.

Accumulation of petroleum hydrocarbon residual considered a major environmental problem in the kingdom of Saudi Arabia cause of intensive efforts for oil detecting. Until now, In situ biodegradation considered the most effective method for petroleum hydrocarbon residual biodegradation. The aim of this study is isolation and identification biodegradable capability bacteria from contaminated sites in Khurais oil field, Dhahran, Saud Arabia via Different morphological and biochemical and molecular methods. Furthermore, degradation level in contaminated liquid medium and soil were evaluated. Three bacterial strains were selected from petroleum-contaminated soils of Khurais oil field depending on their capacity to grow in the existence of hydrocarbon components and identified according to morphological, biochemical. Interestingly, 16S rDNA sequencing fingerprinting results confirmed our bacterial identification as Bacillus subtilis, Pseudomonas aeruginosa and Bacillus cereu. Phyllogenetic tree was constructed and genetic similarity was calculated according to alignments results. Biodegradation patterns for different three isolates were reflected varied degradation ability for three isolates regarding incubation time. Different features were studied for three biodegrading bacterial strains and identified as Bacillus subtilis, Pseudomonas aeruginosa and Bacillus cereus. Remarkable degradation rate % patterns for hydrocarbons residual were recorded for all three isolates with varied.

Transcriptomic and biochemical analysis of upland cotton (Gossypium hirsutum) and a chromosome segment substitution line from G. hirsutum × G. barbadense in response to Verticillium dahliae infection.

BACKGROUND: Verticillium wilt (VW), also known as "cotton cancer," is one of the most destructive diseases in global cotton production that seriously impacts fiber yield and quality. Despite numerous attempts, little significant progress has been made in improving the VW resistance of upland cotton. The development of chromosome segment substitution lines (CSSLs) from Gossypium hirsutum × G. barbadense has emerged as a means of simultaneously developing new cotton varieties with high-yield, superior fiber, and resistance to VW. RESULTS: In this study, VW-resistant investigations were first conducted in an artificial greenhouse, a natural field, and diseased nursery conditions, resulting in the identification of one stably VW-resistant CSSL, MBI8255, and one VW-susceptible G. hirsutum, CCRI36, which were subsequently subjected to biochemical tests and transcriptome sequencing during V991 infection (0, 1, and 2 days after inoculation). Eighteen root samples with three replications were collected to perform multiple comparisons of enzyme activity and biochemical substance contents. The findings indicated that VW resistance was positively correlated with peroxidase and polyphenol oxidase activity, but negatively correlated with malondialdehyde content. Additionally, RNA sequencing was used for the same root samples, resulting in a total of 77,412 genes, of which 23,180 differentially expressed genes were identified from multiple comparisons between samples. After Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis on the expression profiles identified using Short Time-series Expression Miner, we found that the metabolic process in the biological process, as well as the pathways of phenylpropanoid biosynthesis and plant hormone signal transduction, participated significantly in the response to VW. Gene functional annotation and expression quantity analysis indicated the important roles of the phenylpropanoid metabolic pathway and oxidation-reduction process in response to VW, which also provided plenty of candidate genes related to plant resistance. CONCLUSIONS: This study concentrates on the preliminary response to V991 infection by comparing the VW-resistant CSSL and its VW-susceptible recurrent parent. Not only do our findings facilitate the culturing of new resistant varieties with high yield and superior performance, but they also broaden our understanding of the mechanisms of cotton resistance to VW.

Variations in biochemical values for common laboratory tests: a comparison among multi-ethnic Israeli women cohort.

BACKGROUND: Biochemical laboratory values are an essential tool in medical diagnosis, treatment, and follow-up; however, they are known to vary between populations. Establishment of ethnicity-adjusted reference values is recommended by health organizations. AIM: To investigate the ethnicity element in biochemical lab values studying women of different ethnic groups. METHODS: Biochemical lab values (n = 27) of 503 adult Israeli women of three ethnicities (Jewish Ashkenazi, Jewish Sephardic, and Bedouin Arab) attending a single medical center were analyzed. Biochemical data were extracted from medical center records. Ethnic differences of laboratory biochemicals were studied using ANCOVA to analyze the center of the distribution as well as quartile regression analysis to analyze the upper and lower limits, both done with an adjustment for age. RESULTS: Significant ethnic differences were found in almost half (n = 12) of the biochemical laboratory tests. Ashkenazi Jews exhibited significantly higher mean values compared to Bedouins in most of the biochemical tests, including albumin, alkaline phosphatase, calcium, cholesterol, cholesterol LDL and HDL, cholesterol LDL calc., folic acid, globulin, and iron saturation, while the Bedouins exhibited the highest mean values in the creatinine and triglycerides. For most of these tests, Sephardic Jews exhibited biochemical mean levels in between the two other groups. Compared to Ashkenazi Jews, Sephardic Jews had a significant shift to lower values in cholesterol LDL. CONCLUSIONS: Ethnic subpopulations have distinct distributions in biochemical laboratory test values, which should be taken into consideration in medical practice enabling precision medicine.

Diferentes métodos para aislamiento y detección de Salmonella spp. en canales porcinas / Different methods for isolation and detection of Salmonella spp. in pig carcasses

RESUMEN La Organización Mundial de la Salud (OMS) define salmonelosis como una de las enfermedades transmitida por alimentos (ETA) de mayor casuística, ampliamente extendida en todo el mundo. La enfermedad es producida por Salmonella spp. y causa una de las zoonosis más frecuentes y de mayor impacto económico. El hombre adquiere la infección después de la ingestión de alimentos contaminados, aunque también puede transmitirse de persona a persona o por vía fecal-oral. Actualmente, las técnicas micro-biológicas de aislamiento convencional para detección de Salmonella spp. son establecidas por el Código Alimentario Argentino para verificar la aptitud de un producto para consumo, pero éstas requieren de 4 a 5 días para la obtención de un resultado, tiempo que juega en contra para el productor y la conservación de dichos alimentos. Por este motivo en este trabajo se analizan los métodos de diagnóstico tradicional según Normas ISO 6579:2002 con algunas modificaciones, los métodos de inmunoensayo comerciales y la Reacción en Cadena de la Polimerasa técnica (PCR) de detección del gen invA implicado en el proceso de invasión de cepas patógenas. Se analizaron 60 muestras procedentes de canales porcinas destinadas a comercialización. Se detectó un 10% de Salmonella spp. Se pudo determinar que el diagnóstico molecular por PCR posee alta sensibilidad, pero no es alentador el resultado que reflejan los test comerciales inmunocromatográficos ya que queda en evidencia la necesidad de alta carga microbiana para un diagnóstico certero.

ABSTRACT The World Health Organization (WHO) defines salmonellosis as one of the most important foodborne diseases, widely spread worldwide. The disease is produced by Salmonella spp. and causes one of the most frequent zoonoses and of greater economic impact. The infection is acquired after ingestion of contaminated food, although it can also be transmitted from person to person or by fecal-oral route. Currently, conventional isolation microbiological techniques for detection of Salmonella spp. are established by the Argentine Food Code to verify the suitability of a product for consumption. But microbiological techniques require 4 to 5 days to obtain a result, time that plays against the producer and the conservation of such foods. For this reason in this work we analyze the traditional diagnostic methods according to ISO standards 6579: 2002 with some modifications, the commercial immunoassay methods and the Polymerase Chain Reaction technique (PCR) detection of the invA gene involved in the process of invasion of pathogenic strains. Sixty samples from pigs destined for commercialization were analyzed. 10% of Salmonella spp. was detected. It was possible to determine that the molecular diagnosis by PCR has high sensitivity, but it is not encouraging the result that reflect the commercial immunochromatographic tests since it is evident the need of high microbial load for a correct diagnosis.

Evaluation of relationship between dental caries, diabetes mellitus and oral microbiota in diabetics.

BACKGROUND: Diabetes mellitus is a metabolic disease leading to abnormal fat, carbohydrate and protein metabolism. Reduced salivary flow rate caused by hyperglycemia is characteristic mainly for periods of poor metabolic control of diabetes, thereby facilitating the growth of aciduric bacteria and caries-lesion development. The objective of our study was to evaluate the effects of diabetes mellitus on dental caries micro-organisms responsible for caries. MATERIALS AND METHODS: This study was carried out on 60 subjects consisting of 2 groups. The Group A (study group) consisted of 30 subjects with diabetes mellitus and dental caries, and the Group B (control group) consisted of 30 subjects with dental caries but no systemic disease. DFS/dfs index in all subjects was evaluated and compared. Unstimulated salivary flow was collected and levels of Streptococcus mutans were analyzed. RESULTS: It was found that the fasting blood sugar in Group A subjects because of which there was increased streptococcus mutans count and hence high caries index as compared to that of Group B. CONCLUSION: From our study, we could conclude that with increased age, blood sugar levels, DMFT values, dental caries increases in diabetics than in normal (control) subjects and therefore relationship does exist between diabetis mellitus, oral microbiota and dental caries.

Searching whole genome sequences for biochemical identification features of emerging and reemerging pathogenic Corynebacterium species.

Biochemical tests are traditionally used for bacterial identification at the species level in clinical microbiology laboratories. While biochemical profiles are generally efficient for the identification of the most important corynebacterial pathogen Corynebacterium diphtheriae, their ability to differentiate between biovars of this bacterium is still controversial. Besides, the unambiguous identification of emerging human pathogenic species of the genus Corynebacterium may be hampered by highly variable biochemical profiles commonly reported for these species, including Corynebacterium striatum, Corynebacterium amycolatum, Corynebacterium minutissimum, and Corynebacterium xerosis. In order to identify the genomic basis contributing for the biochemical variabilities observed in phenotypic identification methods of these bacteria, we combined a comprehensive literature review with a bioinformatics approach based on reconstruction of six specific biochemical reactions/pathways in 33 recently released whole genome sequences. We used data retrieved from curated databases (MetaCyc, PathoSystems Resource Integration Center (PATRIC), The SEED, TransportDB, UniProtKB) associated with homology searches by BLAST and profile Hidden Markov Models (HMMs) to detect enzymes participating in the various pathways and performed ab initio protein structure modeling and molecular docking to confirm specific results. We found a differential distribution among the various strains of genes that code for some important enzymes, such as beta-phosphoglucomutase and fructokinase, and also for individual components of carbohydrate transport systems, including the fructose-specific phosphoenolpyruvate-dependent sugar phosphotransferase (PTS) and the ribose-specific ATP-binging cassette (ABC) transporter. Horizontal gene transfer plays a role in the biochemical variability of the isolates, as some genes needed for sucrose fermentation were seen to be present in genomic islands. Noteworthy, using profile HMMs, we identified an enzyme with putative alpha-1,6-glycosidase activity only in some specific strains of C. diphtheriae and this may aid to understanding of the differential abilities to utilize glycogen and starch between the biovars.