Standard Dilution Research Articles

Method for the detection and quantification of viral contamination during the preparation of gene therapy drugs in a hospital pharmacy

ObjectiveThe objective of the present study was to develop a method for sampling and detecting an adenovirus-derived gene therapy (GT) vector on isolator worksurfaces.MethodsWe used a quantitative PCR (q-PCR) to...

Fighting H. pylori with Medicinal Plants: A Study on Jordan's Traditional Remedies

Aims of the Study: This study was designed to evaluate the antimicrobial activity of some medicinal plants used among Jordanians for the treatment of gastritis and gastric ulcers against H. pylori. Moreover, plants' inhibitory activity against the H. pylori urease enzyme was also evaluated. Materials and Methods: The activity of 11 medicinal plants used by common people and herbalists to treat ulcers was evaluated against H. pylori (NCTC 11916). Ethanol and essential oil extracts from the tested plants were evaluated using a standard agar dilution method and the MICs were determined. Furthermore, the potential inhibitory effect of each preparation was tested against the enzyme urease using a kinetic colorimetric assay. Results: Cinnamomum cassia oil showed the highest efficiency against H. pylori with the lowest MIC (0.0122 mg.mL-1), followed by Origanum syriacum and Foeniculum vulgare (MICs of 0.39 mg.mL-1). Furthermore, significant urease inhibition activity was recorded for Carum carvi oil (IC50~0.45 mg.mL-1). C. cassia oil (IC50 ~2.8 mg.mL-1), Aloysia citriodora, and Artemisia Judaica (IC50 5.8 mg.mL-1) reported potential urease inhibition activities. Conclusion: Herbs used in Jordanian traditional medicine were found to have anti-H. pylori and significant urease inhibitory activity. These findings might support the use of medicinal plants as adjuvant or alternative therapy for the treatment of H. pylori.

Design and implementation of a portable high-altitude combustible gas alarm detection device

Abstract High-altitude operations present significant safety risks and challenges in accurately detecting combustible gases due to the inherent dangers of working at heights and the technical limitations of traditional detection methods. This study proposes a portable high-altitude combustible gas alarm detection device designed to address the safety risks and accuracy issues associated with high-altitude operations. By integrating wireless communication technology, precise gas concentration control, and automated flow adjustment, this device enables remote ground-based detection, eliminating the safety hazards of high-altitude operations while ensuring measurement accuracy. The device’s real-time monitoring of gas concentration within the calibration hood mitigates the issue of standard gas dilution in the air, and automated control further enhances detection precision. Experimental results demonstrate the device’s suitability for high-altitude alarm calibration, with a concentration measurement error of less than ±5% FS. This research offers a novel technical solution for the on-site detection of high-altitude combustible gas alarms.

Особливості протимікробної дії препаратів полігексаніду до домінуючих збудників ранової інфекції в Україні: сучасні реалії

The treatment of acute and chronic wounds with critical colonization by wound pathogens remains a serious problem, which is becoming more and more threatening every year due to the global problem of antibiotic resistance. Aim - to study the antimicrobial activity of modern antiseptic Polyhexanide and antimicrobial biomaterials based on it against most common pathogens of wound infections. Materials and methods. Antimicrobial activity of polyhexanide solution were studied against MLS -resistant clinical isolates of S. aureus, MDR- strains of E. coli, clinical XDR-isolates of K. pneumoniae, P. aeruginosa, and A. baumannii as well as referent strains S. aureus ATCC 25923, E. coli ATCC 25922, K. pneumoniae ATCC 708603, P. aeruginosa ATCC 27853, A. baumannii ATCC ВАА-747. MIC and MBC of polyhexanide were detected by standard dilution method; bactericidal index of antiseptic activity (BC IAA) for 0.1% polyhexamethylen-biguanide solution for wound irrigation (PHMB) was calculated. The bioactivity of wound antimicrobial dressings with polyhexamethylenguanidyn-hydrochoridi (PHMB-HCl) and PHMB was determined by standard disk diffusion method; growth inhibition zones (GIZ) were measured and compared. Results. MLS-resistant strains of S. aureus were the most susceptible to the action of the antiseptic: the MIC and MBC of PHMB were on average 22.69±4.6 μg/ml and 40.55±8.57 μg/ml, respectively. The MIC values against S. aureus was lower than those against E. coli by 1.97 times, against K. pneumoniae – by 1.78 times, against A. baumannii – by 2.97, against P. aeruginosa – 3.9 times. The MBC of PHMB against clinical strains of S. aureus had the lowest values and differed significantly from those against E. coli by 2.21 times, K. pneumoniae by 1.99 times, A. baumannii by 3.4 times, P. aeruginosa by 4.52 times. The bactericidal index of antiseptic activity (BC IAA) of PHMB had the highest values in relation to clinical strains of S. aureus (BC IAA=24.7), K. pneumoniae (BC IAA=12.4) and E. coli (BC IAA=11.2). Wound dressings PHMB-HCl and PHMB demonstrated high antimicrobial properties against reference and clinical strains of S. aureus, E. coli, K. pneumoniae. Reference and clinical strains of P. aeruginosa were the least susceptible to PHMB-based wound dressings. Conclusions. Polyhexanide antiseptic solution for wound irrigation and polyhexanide-containing dressings with PHMB and PHMB-HCl have the high antimicrobial properties in relation to the most common types of wound pathogens. No conflict of interests was declared by the authors.

Development and validation of a quantitative PCR assay for detection of Sulawesi tortoise adenovirus

In 2007, a mortality event involving over 100 Sulawesi tortoises (Indotestudo forsteni), two Impressed tortoises (Manouria impress) and a critically endangered Burmese star tortoise (Geochelone platynota) was attributed to Sulawesi tortoise adenovirus (STADV; genus Siadenovirus). We developed a TaqMan quantitative PCR assay targeting the DNA polymerase gene of STADV for use in clinical diagnosis and epidemiologic surveillance. This assay failed to amplify five closely-related chelonian adenoviruses, indicating high analytical specificity. The assay performed with high efficiency (slope = −3.337; R2 = 0.999) and high inter- and intra-assay repeatability (coefficient of variation <1.36 % at all standard curve dilutions). Dynamic range included 1.00 × 107 to 1.00 × 101 target copies per reaction and limit of detection was 101 target copies per reaction, though 100 target copies per reaction were intermittently detected. This qPCR assay provides a valuable diagnostic tool for characterization of STADV epidemiology, including potential identification of the North American reservoir host.

Antimicrobial susceptibility of Peruvian Yersinia ruckeri isolates to oxytetracycline and florfenicol using MIC testing.

Yersiniosis, caused by Yersinia ruckeri, has become the most common disease in farmed rainbow trout Oncorhynchus mykiss in Peru, affecting Puno and Junín Regions, important aquaculture areas in the country. Florfenicol (FLO) and oxytetracycline (OXY) are the antimicrobials most frequently used to mitigate losses attributed to this pathogen. This study presents an analysis of the susceptibility patterns of 60 Y. ruckeri isolates (30 isolates each from Puno and Junín), including the type strain CECT 4319T and the strains CECT 955 and CECT 956, against FLO and OXY. Minimum inhibitory concentrations (MICs) were determined following the guideline for standard broth dilution method published by the Clinical and Laboratory Standards Institute. MIC results ranged from 4.0 to 8.0 µg ml-1 for FLO and 0.5 to 4.0 µg ml-1 for OXY. Normalized resistance interpretation (NRI) analysis identified epidemiological cut-off values of ≤16.0 µg ml-1 for FLO and ≤4.0 µg ml-1 for OXY. All Peruvian isolates, including the collection strains, were categorized as wild-type for both antimicrobials. Even though the number of Y. ruckeri isolates with MIC values of 8 µg ml-1 for FLO is more than double in Puno than in Junín (15 vs. 7 isolates), the NRI analysis showed the same epidemiological cutoff of 16 µg ml-1; while for OXY, it was 4.0 µg ml-1 for Puno and 2.0 µg ml-1 for Junín. This study establishes the basis for monitoring susceptibility to FLO and OXY in new Y. ruckeri isolates in Peruvian rainbow trout farming.

Nitazene test strips: a laboratory evaluation

Background2-Benzylbenzimidazole ‘nitazene’ opioids pose a growing threat to public health. Nitazene analogues are increasingly found mixed with or (mis)sold as heroin and in falsified (non-)opioid medications, posing a great risk of intoxication in users (un)knowingly exposed to these potent opioids. Lateral flow immunoassay nitazene test strips (NTS; BTNX Rapid Response™) became commercially available in Q1 2024, with the aim to enable rapid detection of nitazene analogues in drug samples. As only limited independent data is available on the performance of these strips, this lab-based study aimed at evaluating their potential for drug checking applications.MethodsFollowing dilution of drug standards in water, the NTS readouts were analyzed independently by two individuals and by ImageJ. The limit of detection for isotonitazene was determined using two manufacturing lots of NTS. Cross-reactivity with 32 other nitazene analogues was evaluated. Six sourced drug samples were tested to explore the ability of NTS to detect the presence of a nitazene analogue in authentic samples.ResultsThe limits of detection for isotonitazene were 2000 or 3000 ng/mL, depending on the lot. Twenty-four of the 33 tested nitazene analogues cross-reacted with the NTS at concentrations ≤ 9000 ng/mL. Structural analysis indicated that either substitution or removal of the 5-nitro group, or lengthening the linker between the two aromatic rings, generally hampered detection. All six authentic drug samples consistently tested positive, with no observed false negatives.ConclusionsThis study provides a better understanding of the potential of NTS for drug checking purposes. Our findings indicate that NTS can theoretically alert to the presence of most nitazene analogues that have emerged on recreational drug markets. However, ‘desnitazenes’ (lacking the 5-nitro group) may yield false negative results due to low cross-reactivity. Although factors like specificity, lot-to-lot variability, nitazene analogue content in drug samples, solubility, and different testing conditions should be considered, our study results indicate that, at least under the conditions evaluated here (using reference standards and sourced powders), NTS are capable of detecting the presence of a wide range of nitazene analogues. Hence, NTS may alert users of the presence of nitazene analogues in drug samples.

Performance evaluation of the quantitative assay Adenovirus ELITe MGB® Kit on EDTA-plasma, using the ELITe BeGenius® system

BackgroundAdenovirus infections constitute an important cause of morbidity and mortality after hematopoietic stem cell transplantation. Detection and monitoring of adenovirus in EDTA-plasma by real-time quantitative PCR is a sensitive tool for identification and management of patients at risk of a potentially fatal infection. ObjectivesThe aim of this study was to evaluate the analytical and clinical performance of the quantitative Adenovirus ELITe MGB® Kit (ELITechGroup S.p.A.) using the ELITe BeGenius® (ELITechGroup S.p.A.) system and compare the assay to a laboratory-developed quantitative real-time PCR assay. Study designAnalytical sensitivity of the Adenovirus ELITe MGB® Kit was determined by testing serial dilutions of the WHO standard. Detection of adenovirus serotypes was assessed using a panel of 51 serotypes. Clinical sensitivity and specificity were determined by comparing the Adenovirus ELITe MGB® Kit results with the laboratory-developed assay results of 155 retrospective and prospective EDTA-plasma samples from transplant recipients. ResultsThe analytical sensitivity of the Adenovirus ELITe MGB® Kit was at least 54 (1.7 Log) IU/mL and the quantitative results showed a high correlation with the WHO standard (R2 = 0.9978; Pearson) within the range of 1.7 to 6.6 Log IU/mL. All 51 adenovirus serotypes were detected. The clinical specificity and sensitivity for EDTA plasma of the Adenovirus ELITe MGB® Kit were 97.4 % and 99.1 % respectively. ConclusionThe Adenovirus ELITe MGB® Kit performed on the ELITe BeGenius® system is a highly sensitive and specific assay for the detection of adenovirus in EDTA-plasma from transplantation patients.

Combination of Standard Addition and Isotope Dilution Mass Spectrometry for the Accurate Determination of Melamine and Cyanuric Acid in Infant Formula.

In the melamine scandals of the early 2000s, different companies of the dairy industry cheated their products by applying chemical substances to feign a higher content of nitrogen. However, this had a severe toxic impact on the kidney health of consumers. As a result, tremendous effort was put into the prevention of further harm to the public. In the present study, a fast-screening method for the determination of melamine and cyanuric acid in infant formula was developed. While a 1D-LC approach is faster and easier to set up, a 2D-LC approach allows for a more accurate result with better selectivity and sensitivity. For both instrumental approaches, the signal ratio of the isotopologues was crucial and had a dominant effect on the results and the measurement uncertainty. For this reason, the different contributions to the measurement uncertainty were determined experimentally using Matched Standard Addition-IDMS and compared to the Exact Matching Double IDMS.

A FORMAÇÃO EX-CÊNTRICA DE KAMBILI EM HIBISCO ROXO DE CHIMAMANDA NGOZI ADICHIE

Postmodern aesthetics can be understood by its significant deconstructive content and this is observed in the dilution of artistic and literary standards, especially when issues of race, class, gender, or even nation are highlighted. It is through the deconstruction of paradigms that this research is directed, by analyzing the work of the Nigerian writer Chimamanda Ngozi Adichie entitled Hibisco Roxo (2011) in which she narrates the formation process of the teenager Kambili and the shy resistance cultivated in the girl that in the end points out for their possible emancipation. In this way, we analyze how the process of formation and de-formation occurs in its trajectory, which in addition to deconstructing the standard formation of perfectibility propagated by the traditional literary model, the Bildungsroman (Formation Novel), subverts it to the ex-perspective -centric of a girl, Nigerian and suffering from the sad legacies of colonialism and patriarchy. To do this, we will base ourselves on studies on the Bildungsroman and the chronotopic change undergone by the literary genre in the works of Bakhtin (1992), Cristina Ferreira Pinto (1990) and Wilma Patrícia Maas (2000); on the concept of eccentricity we are guided by the studies of literary critic Linda Hutcheon (1988) and on Emancipation we draw on discussions present in Kant (1783) and Adorno (1995).

In vitro antimicrobial resistance of Escherichia coli, Serratia marcescens, Klebsiella oxytoca, and Klebsiella pneumoniae on Bavarian dairy farms between 2014 and 2022

The objective of this study was to describe the prevalence of antimicrobial resistance of E. coli, K. oxytoca, K. pneumoniae, and S. marcescens from quarter milk samples submitted to the udder health laboratory of the Bavarian Animal Health Services (TGD) in Southern Germany between 2014 and 2022. All samples were tested with the California Mastitis Test and analyzed with a standard microbroth dilution to determine the minimum inhibitory concentrations (MIC). The antimicrobials tested were amoxicillin/clavulanate, cefazoline, kanamycin/cefalexin, cefoperazone, cefquinome, and marbofloxacin. Breakpoints were chosen in accordance with CLSI. Over the study period, E. coli, K. oxytoca, and K. pneumoniae showed only few resistances to all antimicrobials tested. For those pathogens MIC 50 and MIC 90 were below breakpoint for all antimicrobials except cefoperazone over the 9 years. A decrease in MIC could be seen for E. coli and K. oxytoca for all of the antimicrobials. While the MIC for K. pneumoniae stayed more stagnant, the prevalence of resistance still decreased overall. S. marcescens isolates were proven intrinsically resistant to amoxicillin/clavulanate and cefazolin and while in vitro resistances were low for all other antimicrobials tested, S. marcescens tended toward higher MIC for most of the antimicrobials over the years. Over time, there was also an overall increase in the number of isolates for all 4 pathogens per year. Starting 2018 there was steep increase in the number of isolates particularly from clinical cases. This jump in numbers coincided with a change of the regulation for veterinary drug prescriptions in Germany in 2018 that required, among other things, antimicrobial resistance testing before a change of antibiotics in the course of treatment and the use of critically important antimicrobials. Overall, while the pathogens increased in numbers, the prevalence of their antimicrobial resistance remained low.

Autocalibration based on dilution of a single concentrated standard is used for the determination of silicate in sea water by the modified molybdenum blue method

The silicate (Si) molybdenum blue method was modified by combining oxalate and ascorbic acid into a single reagent and was used for determining Si in sea water samples. The first step of this automated assay protocol was designed to perform either a calibration by a single Si standard prepared in deionized (DI) water, or to dilute samples in the range of 0–160 μM Si to fit into 0–20 μM Si calibration range using a 20 cm flow cell. By designing the assay protocol to function in batch mode, the influence of salinity on calibration was eliminated, thus making the method suitable for analysis of samples collected in the open ocean, coastal areas, or rivers. Reproducibility and accuracy of this method were evaluated by analysis of certified sea water reference materials. Phosphate (P) does not interfere significantly if the Si:P ratio is 4:1 or larger. The limit of detection was 514 nM Si, r.s.d. 2.1 %, sampling frequency 40 s/h, reagent consumption 700 μL/sample, and using deionized water as the carrier solution.

Phenotypic and genomic characterization of a multidrug-resistant Salmonella enterica serovar Kentucky ST198 isolated from a patient in China

ObjectivesThe objective of this study was to investigate the resistance mechanisms of a multidrug-resistant Salmonella Kentucky ST198 FJ-2064 isolated from a patient in China. MethodsThe antimicrobial susceptibility of FJ-2064 was determined by the standard disc dilution and broth microdilution methods. The complete genome of FJ-2064 was sequenced using PacBio and Illumina MiSeq platforms. Polymerase chain reaction (PCR) and S1-PFGE were utilized to confirm the mutation sites and the genomic plasmids, respectively. ResultsIsolate FJ-2064 belongs to sequence type ST198 and harboured no visible large plasmids, but was concurrent resistant to 22 detected antimicrobial agents including cefotaxime, ciprofloxacin, and azithromycin. The complete genome sequence identified 20 acquired antibiotic resistance genes (ARGs) and five chromosomal mutations in the gyrA and parC genes of the quinolone resistance determining regions (QRDRs) in FJ-2064. In addition, PCR sequencing confirmed that most of the ARGs were clustered on one multidrug-resistant region and a variant of SGI1-K. In particular, the bla-TEM-1 and bla-CTX−M-55, qnrS1, mph(A) genes, which confer resistance to cephalosporins, quinolones, and macrolides respectively, were all located on the multidrug-resistant region. ConclusionsWe have demonstrated one multidrug-resistant region and a variant of SGI1-K in a Salmonella Kentucky ST198 that is co-resistant to cefotaxime, ciprofloxacin, and azithromycin.

Streamlining Protein Fractional Synthesis Rates Using SP3 Beads and Stable Isotope Mass Spectrometry: A Case Study on the Plant Ribosome.

Ribosomes are an archetypal ribonucleoprotein assembly. Due to ribosomal evolution and function, r-proteins share specific physicochemical similarities, making the riboproteome particularly suited for tailored proteome profiling methods. Moreover, the structural proteome of ribonucleoprotein assemblies reflects context-dependent functional features. Thus, characterizing the state of riboproteomes provides insights to uncover the context-dependent functionality of r-protein rearrangements, as they relate to what has been termed the ribosomal code, a concept that parallels that of the histone code, in which chromatin rearrangements influence gene expression. Compared to high-resolution ribosomal structures, omics methods lag when it comes to offering customized solutions to close the knowledge gap between structure and function that currently exists in riboproteomes. Purifying the riboproteome and subsequent shot-gun proteomics typically involves protein denaturation and digestion with proteases. The results are relative abundances of r-proteins at the ribosome population level. We have previously shown that, to gain insight into the stoichiometry of individual proteins, it is necessary to measure by proteomics bound r-proteins and normalize their intensities by the sum of r-protein abundances per ribosomal complex, i.e., 40S or 60S subunits. These calculations ensure that individual r-protein stoichiometries represent the fraction of each family/paralog relative to the complex, effectively revealing which r-proteins become substoichiometric in specific physiological scenarios. Here, we present an optimized method to profile the riboproteome of any organism as well as the synthesis rates of r-proteins determined by stable isotope-assisted mass spectrometry. Our method purifies the r-proteins in a reversibly denatured state, which offers the possibility for combined top-down and bottom-up proteomics. Our method offers a milder native denaturation of the r-proteome via a chaotropic GuHCl solution as compared with previous studies that use irreversible denaturation under highly acidic conditions to dissociate rRNA and r-proteins. As such, our method is better suited to conserve post-translational modifications (PTMs). Subsequently, our method carefully considers the amino acid composition of r-proteins to select an appropriate protease for digestion. We avoid non-specific protease cleavage by increasing the pH of our standardized r-proteome dilutions that enter the digestion pipeline and by using a digestion buffer that ensures an optimal pH for a reliable protease digestion process. Finally, we provide the R package ProtSynthesis to study the fractional synthesis rates of r-proteins. The package uses physiological parameters as input to determine peptide or protein fractional synthesis rates. Once the physiological parameters are measured, our equations allow a fair comparison between treatments that alter the biological equilibrium state of the system under study. Our equations correct peptide enrichment using enrichments in soluble amino acids, growth rates, and total protein accumulation. As a means of validation, our pipeline fails to find "false" enrichments in non-labeled samples while also filtering out proteins with multiple unique peptides that have different enrichment values, which are rare in our datasets. These two aspects reflect the accuracy of our tool. Our method offers the possibility of elucidating individual r-protein family/paralog abundances, PTM status, fractional synthesis rates, and dynamic assembly into ribosomal complexes if top-down and bottom-up proteomic approaches are used concomitantly, taking one step further into mapping the native and dynamic status of the r-proteome onto high-resolution ribosome structures. In addition, our method can be used to study the proteomes of all macromolecular assemblies that can be purified, although purification is the limiting step, and the efficacy and accuracy of the proteases may be limited depending on the digestion requirements. Key features • Efficient purification of the ribosomal proteome: streamlined procedure for the specific purification of the ribosomal proteome or complex Ome. • Accurate calculation of fractional synthesis rates: robust method for calculating fractional protein synthesis rates in macromolecular complexes under different physiological steady states. • Holistic ribosome methodology focused on plants: comprehensive approach that provides insights into the ribosomes and translational control of plants, demonstrated using cold acclimation [1]. • Tailored strategies for stable isotope labeling in plants: methodology focusing on materials and labeling considerations specific to free and proteinogenic amino acid analysis [2].

Systematic evaluation of the influence of Taq enzyme choice and amplification conditions on targeted environmental DNA assay performance and detection in field samples

AbstractTargeted environmental DNA (eDNA) studies mainly rely on quantitative real‐time polymerase chain reaction (qPCR) to amplify extremely low concentrations of DNA present in environmental samples. Understanding factors that influence targeted eDNA assay performance and detection in field samples, such as Taq DNA polymerase enzyme type and thermocycle protocol, is critical for the interpretation of qPCR results. We completed a systematic performance evaluation of five distinct targeted eDNA assays (eANFI6, eFISH1, eANBO5, eGLIN1, and eLICA3 targeting sablefish, general fish, Boreal toad, wood turtle, and the American bullfrog) by subjecting the same samples to analysis by five different Taq enzyme reagent mixes (Immolase, Environmental Master Mix (EMM), Amplitaq, QIAcuity, and QuantiNova) and the commonly used 2‐step (95°C, 60°C) and 3‐step (95°C, 64°C, 72°C) thermocycle protocols. We evaluated assay performance using a standardized dilution series of synthetic dsDNA target sequences and calculated limits of detection (LOD) and quantification (LOQ) and 95% confidence intervals for each combination of Taq enzyme and thermocycle protocol. All assays performed within acceptable performance criteria as defined by the Canadian national standard for targeted eDNA assays. Based on data generated by synthetic dsDNA fragments, the eDNA assays performed comparably regardless of the enzyme reagent mix and protocol used, except for eANFI6 and eGLIN1 using EMM and the 3‐step protocol, where no amplification was observed. On freshwater field samples, EMM and Immolase performed best. On marine field samples, Immolase, EMM, Qiacuity, and QuantiNova performed equally well, although EMM failed to amplify some samples. The work reveals that an enzyme reaction mix or thermocycle protocol can affect the result of an eDNA assay, but the appropriate choice also depends on the nature of the field sample. It is therefore imperative that these are considered when selecting appropriate reaction conditions and that they are clearly reported.

GC-MS analysis of hospital waste soil and isolation of antimicrobial agent producing bacteria

The current study, deals with the GC-Mass analysis of soil and antibiosis activity of soil bacteria, isolated from district hospital at Kabir Chaura, Varanasi, India. Standard serial dilution plate procedures were used to isolate the bacteria. Gram staining was used to identify the isolates morphologically. Strain KC14 was Gram-negative and all other isolates were Gram-positive. Isolated bacteria were tested against six human pathogens viz., Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Staphylococcus aureus, Shigella flexneri and Bacillus subtilis. Primary and secondary screening were done by perpendicular streaking and seed overlay techniques. Based on the results most potential isolates were selected for tertiary screening. The GC-MS of soil samples revealed some major medical pollutants such as para-hydroxy acetanilide, 2-methyl-5-nitro-imidazole and 2-acetoxybenzoic acid. The KC14 isolate was chosen for further studies after demonstrating positive results against S. aureus with a maximum antagonistic activity of 17.3 ± 0.5 zone of inhibition. The molecular identification of KC14 isolate was determined by using 16S rRNA sequencing. The obtained nucleotide sequences were subjected to BLASTN in NCBI, and found similar to the available sequence of Acinetobacter junii. The nucleotide sequences have been deposited in GenBank with accession no. OP265396. The active compound was partially purified by ethyl acetate and tested its activity against S. aureus. This study defines the composition of hospital waste soil and identified a Gram-negative bacterium producing antibacterial activity against important human pathogens.

Экспрессия ARID1a в эндометрии женщин репродуктивного возраста с аномальными маточными кровотечениями и ожирением

Objective. The aim of the study was to evaluate the expression of the proliferative marker ARID1a in endometrial biopsies of patients with abnormal uterine bleeding (AUB) and obesity and its role in the pathogenesis of endometrial dysfunction. Materials and methods. The study included 40 endometrial samples from patients of reproductive age with AUB-E: 20 from patients with AUB-E and obesity (group 1) and 20 from patients with normal body mass index (BMI) and AUB (group 2). 10 endometrial samples were obtained from healthy women of reproductive age, taken on the 2nd-3rd day of the menstrual cycle. All patients underwent histological examination according to the standard procedure. At the second stage, an immunohistochemical study of the ARID1a proliferation marker was performed using polyclonal rabbit antibodies to Anti-ARID1a (clone EPR13501-73) in standard dilution (1:500) according to the protocol of the manufacturer Abcam (Great Britain). The IBM SPSS Statistics v.26 program (IBM Corporation, USA) was used for statistical data analysis. To determine statistically significant differences between quantitative parameters with a normal distribution, one-factor analysis of variance (ANOVA) was used with the calculation of a 95% confidence interval followed by the post-hoc Games-Howell test for multiple comparisons. The differences were considered significant at p&lt;0.05. Results. ARID1a expression was evaluated in the glandular and stromal compartments of the endometrium. In the endometrial glands, the expression of ARID1a was statistically significantly higher in the first group of obese patients compared with the normal and control groups. In the endometrial stroma, the expression of the proliferative marker was higher in the main group compared with the control group. However, when comparing between groups within the main group, statistical significance in the level of expression was not achieved. Conclusion. The results obtained demonstrate the possible pathological effect of obesity on the structural and functional properties of the endometrium, which leads to delayed regeneration of the endometrium due to the glandular component and, consequently, to AUB-E.

Evaluation of Aeris mid-infrared absorption (MIRA), Picarro CRDS (cavity ring-down spectroscopy) G2307, and dinitrophenylhydrazine (DNPH)-based sampling for long-term formaldehyde monitoring efforts

Abstract. Current formaldehyde (HCHO) measurement networks rely on the TO-11A offline chemical derivatization technique, which can be resource intensive and limited in temporal resolution. In this work, we evaluate the field performance of three new commercial instruments for continuous in situ formaldehyde monitoring: the Picarro cavity ring-down spectroscopy G2307 gas concentration analyzer and Aeris Technologies' mid-infrared absorption Pico and Ultra gas analyzers. All instruments require regular drift correction, which is accomplished through instrument zeroing using dinitrophenylhydrazine (DNPH)-coated cartridges, Drierite, or molecular sieves, while heated Hopcalite failed to remove all incoming HCHO. We show that a modified precision estimate accounting for regular instrument zeroing results in values of 0.09, 0.20, and 0.22 ppb at a 20 min integration time for the G2307, Ultra, and Pico, respectively. After applying standard addition and dynamic dilution calibrations, all instruments agreed within 13 % and were well correlated with each other (all r ≥ 0.90). TO-11A HCHO observations resulted in a normalized mean bias of −58 % compared to co-located Picarro G2307 measurements (r=0.62, slope = 0.38, int = 0.07 ppb HCHO). Using a 6-month deployment period in the Atlanta metropolitan area, we determined that the Picarro G2307 and Aeris units have sufficient accuracy and precision to capture the Atlanta spatial HCHO gradient. We find that midday HCHO concentrations have decreased by 22.3 % since 1999 in the city's urban core, and DNPH measurements at a nearby Photochemical Assessment Monitoring Station (PAMS) site show a greater decrease of 53 %.

Association between Physical Activity and Health Outcomes (High Body Fatness, High Blood Pressure) in Namibian Adolescents and Adult women.

Regular physical activity (PA) is known to promote the physical and mental health of children and adolescents and further prevent the development of health problems in adulthood. Information on body composition and PA is crucial for health promotion strategies and for epidemiological studies informing policies. However, there is limited data on the association between body composition and PA in Namibia. This dearth of published data is a significant shortcoming in the development of strategies and policies to promote PA in Namibia. Therefore, this cross-sectional study was conducted to determine the association between PA as a dependent variable and independent variables such as high blood pressure and body fatness as measured by different methods (gold standard deuterium dilution, body mass index, mid upper arm circumference, and waist circumference). The study included 206 healthy adolescent girls aged 13-19 years and 207 young adult females aged 20-40 years from Windhoek, Namibia. PA was measured using the PACE+ questionnaire in adolescents, and the GPAQ questionnaire was used for adults. In adolescents, only 33% of the participants met the recommended guidelines for PA, compared to only 2% for adults. Nevertheless, the study found no statistically significant association between PA and blood pressure indices (p-value < 0.05) among adolescents and adults. However, there was a significant association between PA and high body fatness (p-value < 0.001) and waist circumference (p-value = 0.014) in adolescents. Among adults, PA was significantly related to waist circumference only. In conclusion, failure to meet recommended PA guidelines is strongly associated with abdominal obesity and high body fatness. The knowledge gained from this study may be used by policymakers in the development of strategic policies and interventions aimed at promoting PA as a public priority and improving health outcomes.

Роль ангиогенного фактора в развитии эндометриальной дисфункции у женщин репродуктивного возраста с аномальными маточными кровотечениями и ожирением

The steady increase in the number of patients with abnormal uterine bleeding (AUB) has a negative impact on a woman's reproductive health. Obesity is defined as one of the risk factors for AUB and its role in the development of AUB-M and AUB-O has been studied to date. However, our earlier analysis of the results of histological examination of endometrial biopsies showed that in obese women with AUB, the desquamation phase (AUB-E) is significantly more common in the endometrium than in normal-weight women. One of the possible pathogenetic mechanisms may be abnormal endometrial vascularization due to systemic proinflammatory status and local tissue hypoxia. Aim. To identify the features of the expression of endothelial markers CD34+ and eNOS in endometrial biopsies of patients with AUB and obesity and their role in the pathogenesis of endometrial dysfunction. Materials and methods. A prospective randomized cohort study included 40 patients of reproductive age with AUB-E. Depending on the presence of obesity, women were stratified into 2 groups: 20 patients with AUB-E and obesity (group 1) and 20 normal-weight patients with AUB-E (group 2). 10 healthy women of reproductive age were included as a control group. All patients underwent endometrial biopsy followed by histological examination according to the standard procedure. At the second stage, an immunohistochemical study of CD34+ and eNOS angiogenesis markers was performed using monoclonal mouse antibodies in standard dilution. The IBM SPSS Statistics v.26 program (IBM Corporation, USA) was used for statistical data analysis. Results. CD34+ expression in patients with AUB and obesity was significantly higher than in normal-weight patients with AUB and healthy women (2,5±0,2, respectively; 95% CI 2,1–2,8 vs. 1,8±0,2; 95% CI 1,5–2,2 and 0,5±0,1; 95% CI 0,4–0,6; p1-2=0,03; p1-3&lt;0,001; p2-3&lt;0,001), as well as eNOS expression (respectively 8,7±0,6; 95% CI 7,3–10,0 vs. 4,0±0,5; 95% CI 3–5 and 1,3±0,2; 95% CI 0,9–1,7; p1-2&lt;0,001; p1-3&lt;0,001; p2-3&lt;0,001). Conclusion. Patients with AUB-E and obesity are characterized by increased expression of CD34+ and eNOS in endometrial tissue, which indicates pathological activation of neoangigenesis. Perhaps one of the mechanisms of pathogenesis is the formation of a proinflammatory profile against the background of obesity, which contributes to the pathological hypervascularization of the endometrium. Effective weight loss measures can probably reduce the incidence and recurrence of AUB-E.