Types Of Samples Research Articles

Stability of Gastric Fluid and Fecal Microbial Populations in Healthy Horses under Pasture and Stable Conditions.

Equine gastrointestinal microbial communities vary across the gastrointestinal tract and in response to diet or disease. Understanding the composition and stability of gastric fluid microbiota in healthy horses is a prerequisite to understanding changes associated with the development of disease. The objective of this study was to describe microbial communities in the gastric fluid and feces of healthy horses longitudinally. Horses were maintained on pasture (6 weeks), stabled (5 weeks), then returned to pasture. A consistent forage diet was provided throughout. Native gastric fluid and feces were collected weekly for full-length 16S ribosomal DNA sequencing and microbial profiling analysis. Fewer taxa were identified in the gastric fluid (770) than in the feces (5284). Species richness and diversity were significantly different between sample types (p < 0.001), but not between housing locations (p = 0.3). There was a significant effect of housing and horse on the Bray-Curtis compositional diversity of gastric (p = 0.005; p = 0.009) and fecal (p = 0.001; p = 0.001) microbiota. When horses moved from pasture to stable, the relative proportions of gastric fluid Lactobacillaceae increased and Streptococcaceae decreased, while fecal Firmicutes increased and Bacteriodota decreased. Within each housing condition, there was no significant week-to-week variation in gastric (p = 0.9) or fecal (p = 0.09) microbiota. Overall, these findings support the maintenance of stable gastric and fecal microbial populations under each management condition, providing a basis for further investigation of gastric fluid microbiota in diseases of the foregut.

Activated Carbon and Biochar Derived from Sargassum sp. Applied in Polyurethane-Based Materials Development.

Activated carbon (AC) and biochar (BC) are porous materials with large surface areas and widely used in environmental and industrial applications. In this study, different types of AC and BC samples were produced from Sargassum sp. by a chemical activation and pyrolysis process and compared to commercial activated carbon samples. All samples were characterized using various techniques to understand their structure and functionalities. The metal content of the samples was characterized by using an inductively coupled optical emission spectrometer (ICP-OES). A toxicity test was applied to investigate the effect of AC/BC on organisms, where Sinapis alba seed and Escherichia coli bacteria-based toxicity tests were used. The results revealed that the samples did not negatively affect these two organisms. Thus, it is safe to use them in various applications. Therefore, the samples were tested as fillers in polyurethane composites and, thus, polyurethane-AC/BC samples were prepared. The amounts of AC/BC mixed into the polyurethane formulation were 1%, 2%, and 3%. Mechanical and acoustic properties of these composites were analyzed, showing that by adding the AC/BC to the system an increase in the compression strength for all the samples was achieved. A similar effect of the AC/BC was noticed in the acoustic measurements, where adding AC/BC enhanced the sound adsorption coefficient (α) for all composite materials.

Clinical Test Comparison for Improved Diagnosis of Mucormycosis

Abstract Mucormycosis is a rapidly progressive and invasive fungal infection caused by members of the order Mucorales. Proper treatment relies on a timely diagnosis. However, traditional fungal culture takes several days to weeks and has low sensitivity for recovering Mucorales, as these organisms are sensitive to specimen preparation, including tissue homogenization, and low storage temperatures. More rapid and sensitive diagnostic methods are needed to promptly detect infection. Our laboratory previously developed a real-time PCR assay that detects the most common genera associated with mucormycosis to help improve diagnosis. The assay was analytically validated for a variety of specimen types including serum, tissue (both fresh and FFPE), respiratory secretions and body fluids. Little is known about the agreement between our laboratory developed PCR and other detection methods such as culture or sequencing. We reviewed 6 years of laboratory data to assess overall PCR positivity rate, positivity rate across sample types, and agreement with culture and/or pan-fungal sequencing. We also calculated turnaround time to results to gauge the diagnostic value of Mucorales PCR. Test comparisons with fungal culture and/or pan-fungal sequencing were performed if testing was ordered within 30 days of each other. In our study, we reviewed 3,804 total PCR orders for 2,195 patients. The overall positivity rate of Mucorales PCR orders was 3.81% and varied among different sample types, with fresh tissue and body fluid having the highest positivity rates (18.11% total tissue (33.3% lung tissue) and 10.94% all body fluid (12.5% pleural fluid)) and sputum and CSF having the lowest positivity rates (&amp;lt;1% positivity for both). In a cohort of 428 patients with fungal culture and/or sequencing also performed, 29 patients tested positive for Mucorales by at least one method. PCR detected Mucorales on average 7.83 days sooner than fungal culture (average time to positive result: 3.65 vs. 11.47 days). In patients with culture- or sequencing-confirmed Mucorales infection, PCR had a 100% agreement when comparing shared samples from the same source and collection time. Mucorales PCR was also able to detect infection in 10 (34.5%) other patients who were culture-negative for Mucorales. Our findings suggest that PCR could be a valuable diagnostic tool for suspected mucormycosis, especially tissue invasive disease. PCR increased case detection by 34.5% with a faster turnaround time than culture with lesser cost than sequencing. The main limitation of our study was lack of access to patient information to assess pretest probability of Mucormycosis.

PROTAC’ING CENTROSOME AMPLIfiCATION PROTEINS: TARGETED PROTEIN DEGRADATION AS A NOVEL THERAPEUTIC STRATEGY IN GLIOBLASTOMA

Abstract AIMS Glioblastomas (GBMs) are the most common and aggressive brain tumours, notoriously difficult to treat with universal treatment resistance. A key feature of cancer cells is centrosome amplification (CA), leading to chromosomal instability and worse patient outcomes, including metastasis development and treatment resistance. In this work, I identified CA proteins overexpressed in GBM and aimed to investigate how the targeted degradation of proteins associated with centrosome amplification can influence the growth and survival of GBM cells. METHOD Based on well-established literature data concerning CA-related genes, I evaluated the expression level of these targets in different types of samples from the Tumour Cancer Genome Atlas dataset of GBM patient samples. Following the identification of CA targets differentially overexpressed, to test the hypothesis that CA proteins could be involved in the survival of GBM cells and be potential therapeutic targets, I used a novel drug modality, named Proteolysis Targeting Chimeras (PROTACs) to degrade two CA proteins. PROTACs can quickly destroy specific proteins, potentially overcoming resistance to treatment. Target degradation was confirmed via immunoblotting and cell viability and cell cycle analysis were performed to evaluate the impact on GBM cells. RESULTS Both primary and recurrent GBM samples display high levels of CA gene signature when compared to noncancerous samples, and some CA genes are particularly upregulated in recurrent samples. The PROTACs were effective in degrading both CA targets in GBM cells, leading to a significant decrease in cell viability and G2 cell cycle arrest. CONCLUSION This study presents initial evidence that CA protein degradation could be a promising avenue for GBM research and therapy. Further research is needed to fully assess the translational potential of PROTACs in GBM treatment.

Widespread mutagenesis and chromosomal instability shape somatic genomes in systemic sclerosis

Systemic sclerosis is a connective tissue disorder characterized by excessive fibrosis that primarily affects women, and can present as a multisystem pathology. Roughly 4-22% of patients with systemic sclerosis develop cancer, which drastically worsens prognosis. However, the mechanisms underlying systemic sclerosis initiation, propagation, and cancer development are poorly understood. We hypothesize that the inflammation and immune response associated with systemic sclerosis can trigger DNA damage, leading to elevated somatic mutagenesis, a hallmark of pre-cancerous tissues. To test our hypothesis, we culture clonal lineages of fibroblasts from the lung tissues of controls and systemic sclerosis patients and compare their mutation burdens and spectra. We find an overall increase in all major mutation types in systemic sclerosis samples compared to control lung samples, from small-scale events such as single base substitutions and insertions/deletions, to chromosome-level changes, including copy-number changes and structural variants. In the genomes of patients with systemic sclerosis, we find evidence of somatic hypermutation or kategis (typically only seen in cancer genomes), we identify mutation signatures closely resembling the error-prone translesion polymerase Polη activity, and observe an activation-induced deaminase-like mutation signature, which overlaps with genomic regions displaying kataegis.

In-depth characterization of food and environmental microbiomes across different meat processing plants

BackgroundProcessing environments can be an important source of pathogenic and spoilage microorganisms that cross contaminate meat and meat products. The aim of this study was to characterize the microbiome of raw materials, processing environments and end products from 19 facilities producing different meat products.ResultsThe taxonomic profiles of the microbial communities evolved along processing, from raw materials to end products, suggesting that food contact (FC) surfaces play an important role in modulating the microbiome of final products. Some species persisted with the highest relative abundance in raw materials, food processing environments and/or in the final product, including species from the genera Pseudomonas, Staphylococcus, Brochothrix, Acinetobacter and Psychrobacter. Processing environments showed a very diverse core microbiota, partially shared with the products. Pseudomonas fragi and Pseudomonas sp. Lz4W (in all sample and facility types) and Brochothrix thermosphacta, Psychrobacter sp. and Psychrobacter sp. P11F6 (in raw materials, FC surfaces and end products) were prominent members of the core microbiota for all facilities, while Latilactobacillus sakei was found as a dominant species exclusively in end products from the facilities producing fermented sausages. Processing environments showed a higher amount of antimicrobial resistance genes and virulence factors than raw materials and end products. One thousand four hundred twenty-one medium/high-quality metagenome-assembled genomes (MAGs) were reconstructed. Of these, 274 high-quality MAGs (completeness > 90%) corresponded to 210 putative new species, mostly found in processing environments. For two relevant taxa in meat curing and fermentation processes (S. equorum and L. sakei, respectively), phylogenetic variation was observed associated with the specific processing facility under study, which suggests that specific strains of these taxa may be selected in different meat processing plants, likely contributing to the peculiar sensorial traits of the end products produced in them.ConclusionsOverall, our findings provide the most detailed metagenomics-based perspective up to now of the microbes that thrive in meat, meat products and associated environments and open avenues for future research activities to better understand the microbiome functionality and potential contribution to meat quality and safety.BW5-yJYgBWXLb4-yN7W47nVideo

Towards high throughput analysis using 96-well plate solid phase extraction to determine sedatives and β-blocker residues in food control monitoring

BackgroundVeterinary drugs are widely used in animal production to prevent infections and treat diseases but, this may cause a risk to consumers. Due to the high number of food samples required to monitor yearly, simple, fast, sensitive and selective analytical methods are needed in control laboratories to ensure consumers safety. Nevertheless, many analytical methodologies available in these laboratories include multiple steps and therefore are time-consuming and hinder the analysis throughput requiring significant amounts of solvents and reagents. ResultsThis work developed a 96-well plate solid phase extraction (SPE) method for the extraction of seven sedatives and a β-blocker in animal kidney by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC−MS/MS). The developed method was validated based on Implementing Regulation (EU) 2021/808 in kidney, meat and fish. The performance characteristics of the validation (1–5 μg kg−1) showed good linearity (R2 > 0.998) while decision limits (CCα) were between 1 and 1.2 μg kg−1. Trueness and precision were determined at three levels (n = 7) and the results showed values ranging from 85 to 103 % and from 1 to 9 %, respectively. The feasibility of the method was demonstrated for the residue control requirement established by EU. Method was applied to 201 samples of kidney, meat and fish. SignificanceThis study is the first to present an optimized and validated holistic method for sedatives and β-blocker using 96-well plate SPE and UHPLC−MS/MS. The method showed good performance in kidney, meat and fish samples being an universal method for any species along the same type of sample. The fast, easy, efficient, reliable and universal method showed high throughput and so reduced the analysis time by nine-fold and the required solvent amount by four times fulfilling the green chemistry principals.

Changes in vaginal cytokines concentrations during artificial insemination and natural service in beef heifers

ObjectiveHeifer development is crucial for the optimization of reproductive efficiency in beef production. Heifer development is largely influenced by nutrition. Nutritional status of these heifers can influence immunological responses that are crucial for reproduction. Commercial Angus heifers (n = 9) were utilized, in a pilot study, to elucidate the effects of sampling time (days 0, 7, 14, 21, 28, and 35), pregnancy status, and type of mating on individual cytokine concentrations and cytokine profiles in the vagina following breeding.ResultsCytokine profiles were analyzed using MetaboAnalyst 5.0 and one-way ANOVAs were performed in R Studio to identify differences in individual cytokines based on sampling time, pregnancy status, and type of mating. Cytokine profiles were different (P = 0.05) 7 days after either mating type. Cytokines, IL-1β, IL-17a, MCP-1, and TNFα were different based on the mating type and pregnancy status. Multiple cytokines, IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-17a, VEGFa, and MIP1α, were different between days regardless of pregnancy status. In conclusion, vaginal cytokines differ based on pregnancy status, type of mating, and time which may be indicative of vital pathways that need to be activated for pregnancy.

Parental mentalization and children's externalizing problems: A systematic review and meta-analysis.

Parental mentalization, as the ability to understand mental states (e.g., desires) behind their children's actions, may play a relevant role in the prevention of future externalizing problems. We conducted a meta-analysis to examine the relationship between parental mentalization and children's externalizing problems. Six electronic databases were searched for studies, published in English or Spanish, linking empirically those two variables. Participants included caregivers and children between 0 and 18 years. The filtering process yielded 42 studies with 52 effect sizes. Random-effect analysis revealed higher parental mentalization associated with fewer externalizing problems, with an effect size of r = -.19 (95% CI [-.25, -.13]). Due to high heterogeneity (I2 = 83.750), further analyses were conducted to explore factors affecting such association. Parenting experience and children's developmental stage moderated the relationship, but approaches to operationalize mentalization (MM or PRF), sample type (clinical/at-risk vs. community), and reporting figure (primary caregiver vs. other informants) did not. The study highlights the significance of parental mentalization as a potential contributor to the prevention of externalizing behaviors among infants, children, and adolescents. Our findings may underscore practical implications for equipping caregivers with mentalization skills, helping them to answer appropriately to their children needs.

Aeschynite Group Minerals Are a Potential Recovery Target for Niobium Resources at the Giant Bayan Obo Nb–REE–Fe Deposit in China

With the development of the steel industry, China’s demand for niobium is increasing. However, domestic niobium resources are not yet stably supplied and are heavily dependent on imports from abroad (nearly 100%). It is urgent to develop domestic niobium resources. The Bayan Obo deposit is the largest rare earth element deposit in the world and contains a huge amount of niobium resources. However, the niobium resource has not been exploited due to the fine-grained size and heterogeneous and scattered occurrences of Nb minerals. To promote the utilization of niobium resources in the Bayan Obo deposit, we focused on the mineralogical and geochemical characterization of six types of ores and mineral processing samples from the Bayan Obo deposit, using optical microscopes, EPMA, TIMA, and LA–ICP–MS. Our results show that: (1) the niobium mineral compositions are complex, with the main Nb minerals including aeschynite group minerals, columbite–(Fe), fluorcalciopyrochlore, Nb–bearing rutile, baotite, fergusonite–(Y), fersmite, and a small amount of samarskite–(Y). Aeschynite group minerals, columbite–(Fe), and fluorcalciopyrochlore are the main niobium-carrying minerals and should be the primary focus of industrial recycling and utilization. Based on mineralogical and geochemical investigation, the size of the aeschynite group minerals is large enough for mineral processing. Aeschynite group minerals are thus a significant potential recovery target for niobium, as well as for medium–heavy REE resources. The Nb–rich aegirine-type ores with aeschynite group mineral megacrysts are suggested to be the most significant niobium resource for mineral processing and prospecting. Combined with geological features, mining, and mineral processing, niobium beneficiation efforts of aeschynite group minerals are crucial for making breakthroughs in the utilization of niobium resources at the Bayan Obo.

Direct genome sequencing of respiratory viruses from low viral load clinical specimens using the target capture sequencing technology.

The use of metagenomic next-generation sequencing technology to obtain complete viral genome sequences directly from clinical samples with low viral load remains challenging-especially in the case of respiratory viruses-due to the low copy number of viral versus host genomes. To overcome this limitation, target capture sequencing for the enrichment of specific genomes has been developed and applied for direct genome sequencing of viruses. However, as the efficiency of enrichment varies depending on the probes, the type of clinical sample, etc., validation is essential before target capture sequencing can be applied to clinical diagnostics. In this study, we evaluated the utility of target capture sequencing with a comprehensive viral probe panel for clinical respiratory specimens collected from patients diagnosed with SARS-CoV-2 or influenza type A. We focused on clinical specimens containing low copy numbers of viral genomes. Target capture sequencing yielded approximately 180- and 2,000-fold higher read counts of SARS-CoV-2 and influenza A virus, respectively, than metagenomic sequencing when the RNA extracted from specimens contained 59.3 copies/µL of SARS-CoV-2 or 625.1 copies/µL of influenza A virus. In addition, the target capture sequencing identified sequence reads in all SARS-CoV-2- or influenza type A-positive specimens with <26 RNA copies/µL, some of which also yielded >70% of the full-length genomes of SARS-CoV-2 or influenza A virus. Furthermore, the target capture sequencing using comprehensive probes identified co-infections with viruses other than SARS-CoV-2, suggesting that this approach will not only detect a wide range of viruses but also contribute to epidemiological studies.IMPORTANCETarget capture sequencing has been developed and applied for direct genome sequencing of viruses in clinical specimens to overcome the low detection sensitivity of metagenomic next-generation sequencing. In this study, we evaluated the utility of target capture sequencing with a comprehensive viral probe panel for clinical respiratory specimens collected from patients diagnosed with SARS-CoV-2 or influenza type A, focusing on clinical specimens containing low copy numbers of viral genomes. Our results showed that the target capture sequencing yielded dramatically higher read counts than metagenomic sequencing for both viruses. Furthermore, the target capture sequencing using comprehensive probes identified co-infections with other viruses, suggesting that this approach will not only detect a wide range of viruses but also contribute to epidemiological studies.

Determinants of Intention to Choose Halal Certified Restaurant in West Java

The establishment of halal-labeled restaurants that offer a diverse range of food and beverages is in line with the growing awareness of halal certification. However, the importance of halal certification was not fully known by the public. This study aimed to investigate the potential influence of knowledge, attitude, and religiosity on consumers' purchase interest in halal restaurants in West Java. In addition, certain eateries emphasize the natural and cultural aspects of the region. The varied traditional cuisine, business centers, and the presence of Muslim communities make West Java an attractive destination for those seeking a religiously oriented and convenient dining experience. This study used primary data collected from 100 respondents through a questionnaire distributed via Google Forms. The study utilized purposive sampling, a type of non-probability sampling, and employed multiple linear regression analysis using SPSS version 29 for data analysis, the study was conducted in 3 stages, namely t-test, f-test, and coefficient of determination test. The findings indicate that attitude and religiosity factors influence consumer purchase intention at the halal restaurant. Conversely, knowledge was not proven to significantly affect public purchase intention at the halal restaurant in this study. This research has significant implications for halal restaurant owners. Halal restaurant owners in West Java can better understand the needs and preferences of Muslim consumers, strengthen their market position, and promote the growth of the halal industry in the region.

Inferring post-transcriptional regulation within and across cell types in human testis.

Single-cell tissue atlases commonly use RNA abundances as surrogates for protein abundances. Yet, protein abundance also depends on the regulation of protein synthesis and degradation rates. To estimate the contributions of such post transcriptional regulation, we quantified the proteomes of 5,883 single cells from human testis using 3 distinct mass spectrometry methods (SCoPE2, pSCoPE, and plexDIA). To distinguish between biological and technical factors contributing to differences between protein and RNA levels, we developed BayesPG , a Bayesian model of transcript and protein abundance that systematically accounts for technical variation and infers biological differences. We use BayesPG to jointly model RNA and protein data collected from 29,709 single cells across different methods and datasets. BayesPG estimated consensus mRNA and protein levels for 3,861 gene products and quantified the relative protein-to-mRNA ratio (rPTR) for each gene across six distinct cell types in samples from human testis. About 28% of the gene products exhibited significant differences at protein and RNA levels and contributed to about 1, 500 significant GO groups. We observe that specialized and context specific functions, such as those related to spermatogenesis are regulated after transcription. Among hundreds of detected post translationally modified peptides, many show significant abundance differences across cell types. Furthermore, some phosphorylated peptides covary with kinases in a cell-type dependent manner, suggesting cell-type specific regulation. Our results demonstrate the potential of inferring protein regulation in from single-cell proteogenomic data and provide a generalizable model, BayesPG , for performing such analyses.

Assessing the Effectiveness of Wet Mount and PCR Methods in Diagnosing Trichomonas vaginalis Infections in Endocervical and Urine samples

Trichomonas vaginalis is the cause of trichomonas’s infection, which is referred to be the global sexually transmitted disease (STD) worldwide. This parasite primarily spreads through sexual activity, though it can also spread through intimate. Vaginal inflammation in females causes a variety of clinical symptoms, such as a profusion of greenish-yellow, frothy, foul-smelling discharge, valvar enlargement, itching, and punctate erythematous lesions of the cervix, commonly referred to as "strawberry cervix." There are numerous scenarios in which women may be sick but not exhibit any symptoms. The purpose of the current study is to compare the diagnostic efficacy of polymerase chain reaction (PCR) and wet mount preparation for detecting T. vaginalis in various sample types, including endocervical discharge, endocervical swabs, and urine, of 300 female patients (married and single), ages ranging from 15 to 62, who were referred to Dohuk Maternity Hospital, Shariya Camp, either inside or outside of Dohuk city. Based on the clinical symptoms of the patients, the diagnosis of parasite infection was validated by microscopically confirming the infection in the laboratory using two methods: PCR and direct wet mount preparation. The study findings demonstrated, out of 240 samples, 61 (25.4%) at Dohuk Maternity Hospital and 15 (25%) out of 60 samples in Shariya Camp were contaminated with T. vaginalis. Of the 300 samples, 74 (24.6%) had wet mount preparation infection, and 75 (25.0%) had PCR infection. In the current study, 42 single women had 10 (23.8%) and 258 married women had 66 (25.5%) infected. The age range of 15 to 25 had a higher infection rate (32.8%).

Assessment of radiological contamination due to gold mining in soil and food crops of Babban Tsauni, Gwagwalada, Nigeria.

This work assessed the activity concentrations of 238U(226Ra), 232Th, and 40K and their associated radiological risks due to exposure to soil and consumption of food crops in Babban Tsauni artisanal gold mine, Gwagwalada, Nigeria, using the gamma spectrometry technique. The mean activity concentrations of 226Ra, 232Th, and 40K in the mine soil were 60.2±9.9, 161.4±16.2, and 664.6±138.2, while they were 46.4±4.9, 79.9±39.3, 266.4±185.4 for tubers and 46.9±15.7, 100.5±35.8, 311.4±132.7 (Bq/kg) for grains, respectively. The results reveal that the activity concentrations of radionuclides in all samples exceeded the recommended values set by United Nations Scientific Committee on the Effects of Atomic Radiation (33, 45, and 420 Bq/kg) except 40K, which fell within the acceptable limit in all food crop sample types. Estimated results for radiological hazard parameters, radium equivalent, annual effective dose due to ingestion, and excess life cancer risk were within safe limits, while the annual effective dose due to external gamma radiation in soil and annual gonadal equivalent dose were significantly high in all investigated samples; these call for constant radiological monitoring.

An automated commercial open access assay for detection of Mycoplasma genitalium macrolide resistance.

Azithromycin, a macrolide antibioticum, is the first-line treatment for Mycoplasma genitalium (MG), but resistant MG is an increasing problem. Macrolide resistance-mediated mutations (MRM) has been linked to point mutations in region V of the MG 23S rRNA gene. We have evaluated an open access analyzer (Panther Fusion, Hologic Inc) for detectability of MRM (mutations A2071G and A2072G) and MG wild type (WT) in clinical samples. Also, the agreement of the Panther Fusion assay results with a corresponding established In-house MRM-WT PCR (ABI 7500) was calculated. Left over material from 55 clinical samples positive for MG by the Aptima test (Hologic) based on transcription-mediated amplification (TMA), collected from January to February 2023 in Region Skåne, Sweden, was analyzed. Specific amplification curves were generated for positive controls of MG mutations (A2071G and A2072G) and WT by the Panther Fusion assay. The limit of detection (LOD) was 5.3 copies/mL for WT, 8.1 copies/mL for mutation A2071G, and 81 copies/mL for mutation A2072G. The overall concordance was 91% between the Panther Fusion and the In-house PCR (Kappa 0.621, 95% CI; 0.327-0.914) for detection of WT or MRM in MG-positive clinical samples. The Panther Fusion detected MRM in 20% (11/55) and WT in 62% (34/55) of the samples. The corresponding In-house PCR results were 25% (14/55) and 65% (36/55). In summary, the Panther Fusion assay demonstrated detection of low copy number of MRM and WT of MG. Among clinical samples substantial agreement between the Panther Fusion and In-house PCR results was observed. Integrating MG-analysis (TMA) and MRM-WT assay on the Panther platform could make MRM testing more readily available. However, the Panther Fusion had a lower success rate (82% vs 90%) for macrolide susceptibility testing, hence testing with a complementary method should be considered for samples where neither WT nor MRM MG are detectable.

The Power of Reagent Titration in Flow Cytometry.

Flow cytometry facilitates the detection of multiple cell parameters simultaneously with a high level of resolution and throughput, enabling in-depth immunological evaluations. High data resolution in flow cytometry depends on multiple factors, including the concentration of reagents used in the staining protocol, and reagent validation and titration should be the first step in any assay optimization. Titration is the process of finding the concentration of the reagent that best resolves a positive signal from the background, with the saturation of all binding sites, and minimal antibody excess. The titration process involves the evaluation of serial reagent dilutions in cells expressing the antigen target for the tested antibody. The concentration of antibody that provides the highest signal to noise ratio is calculated by plotting the percentage of positive cells and the intensity of the fluorescence of the stained cells with respect to the negative events, in a concentration-response curve. The determination of the optimal antibody concentration is necessary to ensure reliable and reproducible results and is required for each sample type, reagent clone and lot, as well as the methods used for cell collection, staining, and storage conditions. If the antibody dilution is too low, the signal will be too weak to be accurately determined, leading to suboptimal data resolution, high variability across measurements, and the underestimation of the frequency of cells expressing a specific marker. The use of excess antibodies could lead to non-specific binding, reagent misuse, and detector overloading with the signal off scale and higher spillover spreading. In this publication, we summarized the titration fundamentals and best practices, and evaluated the impact of using a different instrument, sample, staining, acquisition, and analysis conditions in the selection of the optimal titer and population resolution.

Oropharyngeal swab sampling for PRRSV detection in large-scale pig farms: a convenient and reliable method for mass sampling

BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) has significant productivity and economic impacts in swine herds. Accurately determining the PRRSV status at the herd level is crucial for producers and veterinarians to implement strategies to control and eliminate the virus from infected herds. This study collected oropharyngeal swabs (OSs), nasal swabs (NSs), oral fluid swabs (OFs), rectal swabs (RSs), and serum samples continuously from PRRSV challenged pigs under experimental conditions and growing pigs under field conditions. Additionally, OSs and serum samples were collected from individual sows from 50 large-scale breeding farms, and the collection of OSs does not require the sows to be restrained. Ct values of PRRSV were detected in all samples using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR).ResultsIn PRRSV challenged pigs, OSs showed a higher PRRSV-positive rate until the end of the observation period. The Ct values of OSs were significantly lower than those of NSs, OFs, and RSs at 2, 8, 12, 14 and 20 days post-challenge (DPC) (P < 0.05). For growing pigs, the positivity rate of PRRSV in OSs was higher than that in other sample types at 30, 70, and 110 days of age. In sows, 24,718 OSs and 6259 serum samples were collected, with PRRSV-positive rate in OSs (9.4%) being significantly higher than in serum (4.1%) (P < 0.05). However, the Ct values of PRRSV RNA in serum were significantly lower than those in OSs (P < 0.001).ConclusionsThe OSs sample type yielded higher PRRSV-positive rates for longer periods compared to NSs, RSs, OFs and serum samples for PRRSV detection in infected pigs. Therefore, OSs has a good potential to be a convenient, practical, and reliable sample type for implementing mass sampling and testing of PRRSV in large-scale pig farms.

Correlation Between Level of Physical Activity Using GPAQ and Exercise Capacity Using 6 Min Walk Test in Non-Insulin Dependent Type 2 Diabetes Mellitus

Background: This study was designed to find the Correlation Between the Level of Physical Activity and Exercise Capacity in Non-Insulin Dependent Type 2 Diabetes Mellitus. Purpose: To find the Correlation between the Level of Physical Activity and Exercise Capacity in Non-Insulin Dependent Type 2 Diabetes Mellitus. Materials and Methods: Materials required for this study were Countdown timer (or stopwatch), Two small cones, Chair, Worksheet on a clipboard, Sphygmomanometer. It was a correlational study design. The sampling method used was Convenient type of sampling. Sixty subjects with non-insulin dependent type 2 were included in this study. The written informed consent was taken. Informed consent document was signed, retained by the principal investigator and a copy was given to the participant. Global Physical Activity Questionnaire and 6 Minutes walk test was used to study the Physical Activity and Exercise Capacity respectively. All the data obtained was statistically analysed using proper tests. Results: The results showed positive significant (p value &lt; 0.0001) correlation between the level of physical activity and exercise capacity, Physical activity level correlates statistically with distance walked in percentage. Conclusion: Our study concluded that Physical activity level correlate statistically with distance walked in type 2 diabetic patients.

Pengaruh Work Life Balance, Job Satisfaction dan Burnout Terhadap Turnover Intention pada Generasi Z Di PT. Rocket Chicken DIY

This research aims to determine the influence of work life balance, job satisfaction, and burnout on turnover intention. The respondents in this research were employees of Rocket Chicken Companny Generation Z in the Special Region of Yogyakarta.This research was conducted from February to Mart 2024. This research is quantitative research with a total of 100 respondents with a sampling technique using non-probability sampling with purposive sampling type. Data collection uses a questionnaire distributed. The analysis technique uses multiple linear aggression with the help of SPSS Version 23 for Windows.The research results concluded that work life balance had a negative and significant effect on turnover intention. Job satisfaction has a negative and significant effect on turnover intention. Burnoout has a positive and significant effect on turnover intention. Work life balance, job satisfaction, and burnout simultaneously have a significant effect on turnover intention.This research is interesting to carry out because there is an opinion that Generation Z is a generation that does not have a long-term commitment to the company