1st Sample Research Articles

Dysbiosis of the initial stool microbiota increases the risk of developing necrotizing enterocolitis or feeding intolerance in newborns

Several perinatal factors influence the intestinal microbiome of newborns during the first days of life, whether during delivery or even in utero. These factors may increase the risk of developing necrotizing enterocolitis (NEC) by causing dysbiosis linked to a NEC-associated microbiota, which may also be associated with other gastrointestinal problems. The objective of our study was to evaluate the potential risks associated with microbial shifts in newborns with gastrointestinal symptoms and identify the intestinal microbiota of neonates at risk for NEC.During the study period, 310 preterm and term newborns’ first passed meconium occurring within 72 h of birth were collected, and the microbiome was analyzed. We identified the risk factors in the NEC/FI group. Regarding microbiota, we compared the bacterial abundance between the NEC/FI group at the phylum and genus levels and explored the differences in the microbial composition of the 1st stool samples. A total of 14.8% (n = 46) of the infants were diagnosed with NEC or FI. In univariate analysis, the mean gestational age and birth weight were significantly lower in the NEC/FI group (p < 0.001). Prolonged rupture of membranes (PROM) > 18 h, chorioamnionitis, and histology were significantly higher in the NEC/FI group (p < 0.001). Multivariate analysis showed that gestational age (GA), prolonged membrane rupture (> 18 h), and early onset sepsis were consistently associated with an increased risk of NEC/FI. Infants diagnosed with NEC/FI exhibited a significantly lower abundance of Actinobacteria at the phylum level than the control group (p < 0.001). At the genus level, a significantly lower abundance of Streptococcus and Bifidobacterium which belong to the Actinobacteria phylum, was observed in the NEC/FI group (p < 0.001). Furthermore, the NEC/FI had significantly lower alpha diversities (Shannon Index,3.39 vs. 3.12; P = 0.044, respectively). Our study revealed that newborns with lower diversity and dysbiosis in their initial gut microbiota had an increased risk of developing NEC, with microbiota differences appearing to be associated with NEC/FI. Dysbiosis could potentially serve as a predictive marker for NEC- or GI-related symptoms.

A-031 Development of novel ELISAs for total GDF-15 and H-specific GDF-15

Abstract Background To develop sensitive and specific ELISAs for the quantitative measurement of total human GDF-15 and H-specific (H202D mutant) GDF-15 in human serum, and other biological fluids. Growth/differentiation Factor 15 (GDF-15) is a divergent member of the TGF-β superfamily of growth factors. It is encoded in humans by a gene in chromosome 19. The human GDF-15 gene encodes for a protein of 308 amino acid residues which consists of a signal sequence (residues 1-29), pro-domain (30-194), and mature growth factor domain (195-308). The molecular weight of a mature GDF-15 dimer is 25 kDa. Approximately 25% of humans have a missense polymorphism in the GDF-15 gene resulting in mutation of histidine 202 to aspartate (histidine 6 in the mature domain), close to the N-terminus of the mature growth factor. This variant is associated with phenotypes in prostate cancer, hyperemesis gravidarum, (severe morning sickness in pregnancy), and rheumatoid arthritis. Methods Highly specific and reproducible total GDF-15 (AL-1014-r) and H-specific GDF-15 (AL-1018-r) ELISAs have been developed using specific monoclonal antibodies to help estimate the total and mutant (DD) concentration in serum in the respective assays. The ELISAs use 10 uL sample volume and are calibrated to recombinant human GDF-15 from R &amp; D Systems (Biotechne, USA). These ELISAs were validated for their specificity using HH, DD, HD recombinant preparations and their circulating levels in male, female, 1st, and 2nd-trimester serum samples. Results The Total GDF-15 ELISA assay detects total GDF-15 including histidine 202 to aspartate mutation (HH, HD &amp; DD variant) in the mature domain in equimolar proportions. The H-specific GDF-15 assay is specific to histidine at 202aa in the GDF-15 sequence and does not detect histidine 202 to aspartate mutation (DD variant). Median Levels of total GDF-15 in healthy males (n=18), females (n=17), 1st trimester (n=20), and 2nd trimester (n=20) were 847.9 pg/mL, 1182.4 pg/mL, 12759.7 pg/mL, and 12951.2 pg/mL. Median Levels of H-specific GDF-15 in healthy males (n=18), females (n=17), 1st trimester (n=20), and 2nd trimester (n=20) were 723.2 pg/mL, 641.3 pg/mL, 8381.4 pg/mL and 4652.1 pg/mL. Method comparison between total GDF-15 and commercial GDF-15 assay using serum samples (HH, wild type) in the range of 400-2500 pg/mL yielded a slope of 0.98 (r=0.97). The HD and DD mutant samples in the total assay recovered at twice the concentrations of the commercial assay. The total and intact assays are highly reproducible with the total coefficient of variation less than 7%. Conclusions Use of total GDF-15 assay in combination with the GDF-15 H-specific assay (DD nondetectable) ELISA can accurately estimate the mutant (DD) concentration in serum. This will enhance the knowledge of the GDF-15 measurements in the literature as the commercial assays were compromised for its immunoreactivity to the HD DD genotypes which is highly prevalent.

Assessment of Antibody Titers after 6 Months of Vaccination against SARS-COV-2 in Patients with CKD Stage 4, 5 and CKD 5d

Background: Since the pandemic of COVID-19 started from December 2019, remarkable numbers of infections and deaths associated with COVID-19 have been recorded worldwide. Chronic kidney disease patients are particularly at high risk of infections due to impairments in the innate and adaptive immune systems. Adequate humoral (antibody) and cellular (T cell-driven) immunity are required to minimize pathogen entry and promote pathogen clearance to enable infection control. Vaccination can generate cellular and humoral immunity against this specific pathogen. COVID-19 prevention through successful vaccination is therefore paramount in chronic kidney disease population. But vaccination efficacy is diminished in these patients because premature ageing of the immune system and chronic systemic low- grade inflammation are the main causes of immune alteration in these patients. Therefore, it is urgently necessary to establish a different vaccination strategy for chronic kidney disease and dialysis patient in terms of the dose and administration time. Aims: This study aimed to assessment of antibody titers after vaccination against SARS-COV-2 in patients with chronic kidney disease stage 4, 5 on conservative management and maintenance haemodialysis. Methods: This prospective observational comparative was conducted in Nephrology department of Dhaka Medical College Hospital. Selectionof patients was done by purposive sampling according to inclusion and exclusion criteria. Total 135 patients distributed in three groups: 45 patients of chronic kidney disease (CKD) stage 4, 5 on conservative management, 45 patients on maintenance haemodialysis (MHD) and 45 healthy controls were approached for the study who were receiving SARS-COV-2 vaccination. Demographic, clinical and laboratory data were collected initially. At first a pre vaccination sample or 1st sample was taken for antibody measurement. Then participants from all groups were given 2 doses MODERNA vaccine containing 100 µg in 0.5 ml each in 28 days apart. Then after 14 days of 1st dose of vaccination the 2nd samples were taken, 3rd samples were taken 14 days after the 2nd dose vaccination. Study populations were subdivided into two groups according to pre vaccination SARS-COV-2 antibody titer; seropositive- positive response before vaccination and seronegative- negative response before vaccination. They were also divided into two groups according to quantitive antibody response; positive response- values ≥10 DU/mL were positive Negative response- values of &lt;10 DU/mL were negative. Result: Seroconversion rate was around 20% among study participants before vaccination. 14 days after the 1st dose of vaccination, 90.04% patients had positive immune response in CKD stage 4, 5 on conservative management group whereas in MHD group 84.82% responded to vaccination and immune response in control group was 100%. Immune response is 100% among all the groups after 14 days of 2nd dose of vaccination but the concentration of antibody differs significantly among the study groups. Responders were comparatively younger with normal BMI. Conclusion: Haemodialysis patients as well as patients with chronic kidney disease stage 4, 5 on conservative management showed a favorable but profoundly lower early antibody response, which decreased substantially during follow-up.

Ineffectiveness of carbapenems in real-world clinical practice according to therapeutic drug monitoring data and Roszdravnadzor AIS reports

Relevance. The use of carbapenems is associated with significant variability in pharmacokinetics/pharmacodynamics (FC/PD) parameters, particularly in critically ill patients. The combination of variability in these parameters and standardized dosing regimens can lead to irrational dosing, excessively high or low doses, and consequently less effective treatment and resistance. Optimal management of these factors is essential for combating the development of resistance, particularly for reserve antibiotics.The aim of this study was to evaluate the achievement of the target levels of carbapenems (imipenem/cilastatin) in plasma concentrations in patients with burn injury based on therapeutic drug monitoring and to analyze spontaneous reports registered in the AIS Roszdravnadzor database regarding the effectiveness of therapy.Methods. The analysis included patients receiving antibiotic therapy with imipenem/cilastatin in the burn unit for adults at the University Hospital of the Volga Region Research Medical University for burn trauma who were hospitalized from 01.03.2023 to 30.06.2023. The study was conducted without correcting the trade name of imipenem/cilastatin. Therapeutic drug monitoring was performed after the 4th administration of imipenem/cilastatin. Blood was drawn at 3 h (1st sample), 5 h (2nd sample), and 8 h (3rd sample) after infusion in a clotting activator tube. The analysis was performed using a liquid chromatograph "LC-20 Prominance" (Shimadzu, Japan) in reversed-phase mode with a matrix photodiode detector for UV and visible spectra. Data processing was performed using the LCsolution program. Spontaneous reports regarding the use of carbapenems recorded in the AIS of Roszdravnadzor from January 2020 to November 2023 were also analyzed as the object of the study.Results. The results of the study of carbapenems (imipenem/cilastatin) concentration level achievement in the plasma of patients with burn injury showed that the effective imipenem concentration exceeding the MPC value in relation to the isolated Gram-negative pathogen was found only in 1 out of 5 patients included in the study. Two patients showed dynamically changing IPC values during treatment, indicating the necessity of therapeutic drug monitoring, as well as the probability of failure to achieve target concentrations and optimal FC/FD values. In another two patients, imipenem concentrations were insufficient to maintain optimal FC/PD values, indicating that the antimicrobial regimen was ineffective. An analysis of spontaneous reports registered in the AIS of Roszdravnadzor regarding identified cases of the ineffectiveness of carbapenems (imipenem/cilastatin, meropenem) revealed 5,2% of reports regarding meropenem and 1.4% regarding imipenem/cilastatin in the total structure of reports.Conclusion. The implementation of therapeutic drug monitoring procedures can reduce therapy ineffectiveness and antibiotic resistance through personalized antimicrobial dosing.

Ambient exposure to fine particulate matter with oxidative potential affects oxidative stress biomarkers in pregnancy.

Prenatal exposures to ambient particulate matter (PM2.5) from traffic may generate oxidative stress, and thus contribute to adverse birth outcomes. We investigated whether PM2.5 constituents from brake and tire wear affect levels of oxidative stress biomarkers (malondialdehyde (MDA), 8-hydroxy-2'-deoxyguanosine (8-OHdG)) using urine samples collected up to three times during pregnancy in 156 women recruited from antenatal clinics at the University of California Los Angeles. Land use regression models with co-kriging were employed to estimate average residential outdoor concentrations of black carbon (BC), PM2.5 mass, PM2.5 metal components, and three PM2.5 oxidative potential metrics during the 4-weeks prior to urine sample collection. 8-OHdG concentrations in mid-pregnancy increased by 24.8% (95% CI: 9.0, 42.8) and 14.3% (95% CI: 0.4%, 30.0%) per interquartile range (IQR) increase in PM2.5 mass and BC, respectively. The brake wear marker (barium) and the oxidative potential metrics were associated with increased MDA concentration in the 1st sample collected (10-17 gestational week), but 95% CIs included the null. Traffic-related air pollution contributed in early to mid-pregnancy to oxidative stress generation previously linked to adverse birth outcomes.

Elecsys® AMH assay: Determination of Anti-Müllerian hormone levels and evaluation of the relationship between superovulation response in Holstein dairy cows.

Anti-Müllerian Hormone (AMH) serves as a crucial parameter in assessing the reproductive herd life and ovarian reserve in cattle. Consequently, extensive research is conducted on AMH levels. Various measurement methods can be employed to determine AMH levels. However, to our knowledge, no study has been conducted on Holstein donors using the Elecsys® AMH kit. This study was designed to determine AMH levels in donors utilising the Elecsys® AMH kit and to evaluate the relationship between superovulation response parameters and AMH levels. In this study, we measured the serum AMH levels of 36 cows using the Elecsys® AMH automated assay before the superovulation protocol (1st sample) and FSH injections (2nd sample). The cows were categorised into three groups based on their AMH levels: low, medium, and high AMH. Positive correlations were identified between AMH and parameters associated with superovulation response. The high AMH level group exhibited significantly greater numbers of corpus luteum, total embryos, transferable embryos, and grade 1 embryos compared to the medium and low AMH groups (p < 0.05) There was no significant difference between AMH levels before the superovulation protocol and FSH injections(p > 0.05). Body condition score and parity did not significantly affect AMH levels in cows (p > 0.05). Also, AMH cut-off values for the number of corpus luteum, total embryo, and transferable embryoswere detectedas 234, 227, and 210 pg/mL, respectively. These findings demonstrate that a high serum AMH level has a positive influence on the superovulation response. AMH can be used as a reliable marker for the selection of donors in Holstein cows.

SPECIES CHARACTERISTICS OF MICROBIAL ASSOCIATIONS IN THE COURSE OF TREATMENT OF 10 TO 14-YEAR-OLD CHILDREN WITH PLAQUE-INDUCED GINGIVITIS

Aim: Еvaluation of the species characteristics of subgingival biofilm microbial associations during treatment of children with plaque-induced gingivitis aged 10-14 years. Materials and methods: The subject of the study were 34 children with plaque-induced gingivitis. Three samples of subgingival biofilm are taken per patient: 1st sample - at the first visit; 2nd sample - two weeks later; 3rd the sample - one month later. Five teeth per child are selected from which a sample is taken for PCR testing for 9 control strains of subgingival microorganisms (A. actinomycetemcomitans, P. gingivalis, T. denticola, T. forsythia, P. intermedia, P. micros, F. nuleatum, E. nodatum, C. gingivalis). Results: C. gingivalis (green complex) is isolated in all examined children before the start of treatment and is indispensably observed in microbial associations with P. intermedia (orange complex). In other cases, associations with 7-8 microorganisms are observed and in a more complex composition. One month after the start of treatment, no periodontopathogens or only C. gingivallis as a single microorganism was is isolated in more than 55% of the samples examined. Microbial associations are predominantly 2 to 4 microorganisms.

A Comparative Study on the General and Otolaryngological Manifestations of COVID-19 in the Hospitalized Population of the Telangana Region During the First and Second Waves.

Introduction Some common symptoms of coronavirus disease 2019 (COVID-19) are fever, cough, and shortness of breath. But ear, nose, and throat (ENT) manifestations such as loss of smell and taste are also very common. Objectives To compare the general and otorhinolaryngological manifestations of COVID-19 and to compare the treatments given and mortality rate during its two waves. Methods This retrospective study was conducted on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cases in a tertiary care teaching hospital. Six hundred patients were included in the 1st wave sample and 516 were in the 2nd wave sample. The data collected included demographics, comorbidities, general, and ENT symptoms, need for ventilatory support, oxygen therapy, and mortality for both the waves. Results Fever, malaise, and myalgia were more frequently presented in the first wave than in the second, whereas shortness of breath was more common in the second wave. In the second wave, a significant increase in anosmia cases was reported, whereas sore throat, nasal obstruction, dysphagia, nasal discharge, and sneezing were significantly reduced compared with the first wave ( p < 0.001). The case fatality rate increased from 11.33 to 21.55% ( p < 0.001) from the 1 st to the 2 nd wave. The patients who died in the second wave were younger than those in the first wave. Two doses of vaccination showed protection from the death over those not vaccinated and those who only received one dose ( p < 0.05). Conclusion Ear, nose, and throat (ENT) manifestations are very common along with the general symptoms. As anosmia and dysgeusia are early presenting symptoms in COVID-19 patients, all physicians should screen patients for ENT symptoms.

Исследование факторов риска развития катаракты нерадиационной природы у лиц, подвергшихся хроническому радиационному воздействию

Relevance. Epidemiological data have revealed a significant association between exposure to ionizing radiation, loss of lens optical function, and cataract formation. Cataract is a multifactorial disease that is traditionally associated with age, exposure to ultraviolet radiation, diabetes, smoking, alcohol abuse, obesity, steroid hormones, hypertension, etc. The chronic nature of the effects of radiation on the body, including both cell damage and regenerative processes, complicates the issue of assessment of the nature and magnitude of the dose-dependence of effects. Purpose. To investigate the potential risk factors of cataract development in different morphological layers, depending on social conditions and several diseases (non-radiation factors) in the population affected by two radiation accidents (dumping of radioactive waste into the Techa River and the East Ural Radioactive Trace) in the Southern Urals. Material and methods. The data from an in-depth examination of patients by an ophthalmologist in the Clinical Department of the URCRM were analyzed. The study involved 1377 patients. The «case» and «control» groups, comparable in sex and age, were selected from the general population. The impact of each factor was assessed on the basis of the Odds Ratio (OR) indicator. Risk factors associated with the development of cataracts in different layers of the lens included arterial hypertension, body mass index, age at the onset of exposure, level of education, alcohol consumption, smoking, ethnicity. Results. An increased risk of developing cataracts in the lens nucleus (1st sample: OR=0.54 (1st sample: 95% CI: 0.34–0.86; 2nd sample: OR=0.51 (95% CI: 0.32–0.82) (p&lt;0.05)) and the intensity of its color (1st sample: OR=0.48 (95% CI: 0.29–0.80); 2nd sample: OR=0.56 (95% CI: 0.34–0.94) (p&lt;0.05)) was established in persons who were at the age over 6 at the onset of exposure. Conclusion. The analysis did not reveal a statistically significant relationship of changes in the cortical layers, the nucleus and the posterior capsule of the lens with other studied risk factors in persons exposed to chronic radiation at a low to medium dose range. There was an increased risk of cataract development in the lens nucleus and the intensity of its color in persons who began to be exposed at the age of over 6 years. Key words: risk factors, cortical cataract, nuclear cataract, posterior subcapsular cataract, chronic exposure

P11.19.A MACROPHAGES-BASED IMMUNOTHERAPY OF SOLID TUMORS MICROENVIRONMENT: THE TEM-GBM STUDY (NCT03866109)

Abstract BACKGROUND We have developed an HSC-based platform to switch from pro-tumor to anti-tumor bone-marrow derived tumor-infiltrating macrophages. Specifically, Temferon cell therapy delivers IFN-a into the tumor microenvironment (TME) in a targeted manner via Tie-2 expressing monocytes, with transcriptional and post-transcriptional controls mediated by miRNA target sequences. MATERIAL AND METHODS TEM-GBM is an open-label, Phase 1/2a dose-escalation study evaluating the safety and efficacy of Temferon in up to 27 newly diagnosed, unmethylated MGMT glioblastoma (GBM) patients. Twenty-four patients are assigned to 8 cohorts and will receive a single increasing dose of Temferon, 3 additional patients will be further enrolled to confirm the safety profile of the chosen Temferon dose to be tested in a broader group of patients (Phase 2). Patients are followed up to 2 years after treatment with Temferon. Here we report the analysis on the biological activity of Temferon on patients who underwent a second surgery intervention. RESULTS As of 31st March 2023, 6 out of 19 Temferon treated patients underwent a second surgical intervention; of these, 4 belonged to low dose cohorts (C1-C2) and 2 to high dose cohorts (C4-C5). The second surgery occurred at a median of 11 months after 1st surgery (range 7-17 months). Where enough fresh material was collected to enrich for CD45+ cells within the tumor, a 3% to 5% of gene-marked tumor infiltrate was found (2/2 specimens) suggesting that Temferon-derived cells homed to the TME. To verify if the local delivery of IFN-a by TEMs was able to upregulate the Interferon Stimulated Genes (ISGs) in tumor tissue, the transcripts for IFIT1, IRF7, OAS1a genes were evaluated by RT-PCR in formalin-fixed paraffin sections obtained at first and second surgery. The analysis was performed on eighteen (18) 1st surgery samples (from C1 to C6), and on six (6) 2nd surgery specimens. At 1st surgery, transcripts were detected in 17 out of 17 specimens (one sample excluded since housekeeping genes did not amplify) and most of the patients showed a relatively low quantity of ISGs expression. The comparison of the overall ISGs expression at first surgery and second surgery highlighted the activation of IFN-responsive genes at second surgery suggesting the occurrence of the local delivery within the TME of biologically active IFN-a. CONCLUSION The results provide initial evidence of Temferon’s potential to modulate the TME of GBM patients.

A descriptive analysis of first trimester urinary concentrations of 14 bisphenol analogues in the MIREC Canadian pregnancy cohort

BackgroundConcern over the health effects of BPA, particularly for the developing fetus, has led to an increasing use of bisphenol analogues in industrial and consumer products, which may be as hormonally active as BPA. Biomonitoring data for many bisphenol analogues, especially in pregnant populations, are limited. MethodsWe measured concentrations of 14 bisphenol analogues in 1st trimester urine samples (n = 1851) from the Maternal-Infant Research on Environmental Chemicals (MIREC) Canadian pregnancy cohort (2008–2011). We examined patterns of exposure according to sociodemographic and sampling characteristics as well as occupation and frequency of consumption of canned fish within the previous 3 months. ResultsBPA was detected in 89% of participants with a specific gravity standardized geometric mean concentration of 0.990 μg/L. Biphenol 4,4’ (BP 4,4′), 4,4′-dihydroxydiphenyl ether (DHDPE), and bisphenol E (BPE) were detected in >97% of participants. Bisphenol F (BPF) and bisphenol S (BPS) were detected in >60% of participants. Specific gravity standardized geometric mean concentrations of these 5 compounds ranged from 0.024 to 0.564 μg/L. Nine bisphenol analogues were detected in <9% of participants. Concentrations of BP 4,4′, DHDPE, and BPE were higher in younger women and those with higher pre-pregnancy BMI, lower household income, lower education, and among smokers. We found a similar pattern of differences in BPF for age, education, and smoking status while BPS similarly differed across categories of pre-pregnancy BMI. Participants who were unemployed or working in the service industry had higher molar sum of 7 bisphenol analogues than those working in healthcare, education, or an office setting. Canned fish consumption was not related to bisphenol analogue concentrations. ConclusionBP 4,4’, DHDPE, BPE, BPF, and BPS were highly detected in 1st trimester urine samples in this large pan-Canadian pregnancy cohort. This suggests widespread exposure to these analogues around 2008–2011 and warrants further investigation into associations with health outcomes.

Measurement of 24 phthalate metabolites in 1st trimester urine samples: The MIREC study

Phthalates are non-persistent chemicals measured as metabolites in urine. Over time, new metabolites have been identified. In the original Maternal-Infant Research on Environmental Chemicals (MIREC) study (2008–2011), we measured 11 phthalate metabolites in first trimester urine samples. The goal of the present study was to develop a method to measure new metabolites, to increase the sensitivity for some previously measured metabolites, and to measure these new metabolites in biobanked urine samples from MIREC participants.Using Ultra Performance Liquid Chromatography with a tandem mass spectrometer, we developed a method to measure 24 metabolites from 10 different parent phthalates. Chromatographic interpretation of some of the di-iso-decyl phthalate metabolites (mono-(2-propyl-6oxoheptyl) phthalate (MOiDP), mono-(2,7-methyl-7-carboxyheptyl) phthalate (MCiNP), mono-(2-propyl-6-hydroxy-heptyl) phthalate (MHiDP)) and di-iso-nonyl phthalate metabolites (mono(oxo-isononyl) phthalate (MOiNP), mono(carboxy-isooctyl) phthalate (MCiOP), mono(hydroxy-isononyl) phthalate (MHiNP) and mono-isononyl phthalate (MiNP)) was challenging as these are complex isomeric mixtures.To validate and confirm our quantitation peaks, an assay using a high-resolution detection technique was developed on a Quadrupole Time-of-Flight (QToF) system. This system has a mass resolution of at least 0.005 amu, compared to 0.5 amu for the MS/MS detector. Using the QToF system, the distinction between an isomer and possible interference was achieved with the use of the exact mass.In about 1800 MIREC samples, mono-cyclo-hexyl phthalate (MCHP), mono-(7-carboxy-n-heptyl) phthalate (MCHpP), mono-iso-decyl phthalate (MiDP), and mono-n-octyl phthalate (MnOP) were rarely detected, while detection of MMP was improved. MCiOP, MiNP and MCiNP had to be reported semi-quantitatively.Given the complexity of isomeric mixtures of some phthalates, researchers must be careful in their determination of the analytes and the approach used in their quantification when generating biomonitoring data.This study produced biomonitoring data for a large population of pregnant people that can be used in risk assessment of phthalates. Future work will examine associations with birth and child outcomes.

SARS-CoV-2 RNAemia and Disease Severity in COVID-19 Patients

Objective: The clinical implications of SARS-CoV-2 RNA viremia in blood (RNAemia) remain uncertain despite gaining more prognostic implications for coronavirus disease 2019 (COVID-19). However, the clinical relevance of SARS-CoV-2 RNAemia has not been well documented. Methods: We conducted a cohort study on 95 confirmed COVID-19 patients and explored the prospects with evidence of SARS-CoV-2 RNAemia in association with various clinical characteristics. We performed reverse transcription-polymerase chain reaction and studied the risk factors of SARS-CoV-2 RNAemia using logistic regression analysis. Results: The presence of SARS-CoV-2 RNAemia in critical or fatal cases was the highest (66.7%), followed by severe (12.5%) and mild to moderate (1.7%) in admission samples. SARS-CoV-2 viral RNAemia was detected on admission and 1st week samples; however, RNAemia was not detected on the samples collected on the second week post-symptom onset. Multiple regression analysis showed that the severity of the disease was an independent predictor of RNAemia (p &lt; 0.021), and the Kaplan–Meier survival curve estimated an increased mortality rate in SARS-CoV-2 RNAemia cases (p &lt; 0.001). Conclusions: Our study demonstrated that SARS-CoV-2 RNAemia is a predictive risk factor for clinical severity in COVID-19 patients. Hence, we showed that blood RNAemia might be a critical marker for disease severity and mortality.

P-053 Sperm aneuploidy and DNA integrity pre- and post-chemotherapy in men with testicular cancer

Abstract Study question Are fragmentation levels (single-double stranded DNA breaks) or aneuploidy frequencies in sperm increased in patients with testicular cancer who recovered spermatogenesis after chemotherapy (CT)? Summary answer Similar levels of sperm DNA fragmentation and sperm aneuploidy frequency are found in men with testicular cancer pre- and post-chemotherapy. What is known already Testicular cancer is the most frequent solid tumor among young men. Therefore, the restoration of fertility and achievement of fatherhood in survivors have become important concerns. The integrity of genetic material in sperm is crucial since some paternal genes are essential for early embryo development. Although previous studies analyzed the effect of CT on sperm DNA integrity, they did not differentiate single vs. double-stranded DNA breaks. Few data are available on aneuploidy in sperm from testicular cancer survivors, so it remains to be elucidated whether cancer treatments may lead to an increased risk of aneuploidy syndromes in offspring. Study design, size, duration Prospective multicenter study including 31 males with a history of testicular cancer who requested semen cryopreservation between 2007-2021. All patients included underwent CT and recovered spermatogenesis once treatment ended. Post-CT samples were collected and seminal parameters, single-double stranded DNA breaks and the proportion of sperm aneuploidies were compared with one aliquot of the initial ejaculate. The project was approved by an Institutional Review Board and written informed consent for participation was obtained from each subject. Participants/materials, setting, methods Patient´s age prior CT was 31.23yrs (IC 95%: 29.29-33.17). Mean interval time between 1st and 2nd samples was 4.53yrs (IC 95%: 3.42-5.64). TUNEL quantified overall sperm DNA fragmentation by flow cytometry analyzing at least 20,000 cells, while γH2AX staining was applied for double DNA fragmentation assessment. Sperm aneuploidies were calculated by using fluorescence in situ hybridization (FISH) probes for chromosomes 13, 18, 21, X, Y. A paired t-test was used to compare variables pre- and post-CT. Main results and the role of chance Similar results were found when sperm quality parameters were compared. Mean sperm concentration pre-CT was 33.03M/mL (IC 95%: 23.98-42.07) and 36.41 M/mL (IC 95%: 27.85-44.96) in post-CT ejaculates (N.S.). A comparable rate of motile spermatozoa was observed in pre-CT vs. post-CT samples [(43.81% (IC 95%: 38.42-49.19) vs. 44.73% (IC 95%: 38.95-50.51), N.S.]. Regarding TUNEL assay results, the sperm DNA fragmentation rate was 21.65% (IC 95%: 17.66-25.63) in pre-CT samples and 23.00% (IC 95%: 19.90-26.61) in post-CT samples (N.S.). Furthermore, similar results were obtained when double-stranded fragmentation levels (H2AX) were compared between pre and post-CT [10.2% (IC 95%: 6.45-13.96) vs. 12.44% (IC 95%: 8.75-16.13) N.S.]. Comparable sperm disomic proportions were found in PRE and POST-CT samples: [(Cr.13 =0.02%, Cr.18 =0.01%, Cr.21=0.30%, Cr.XY=0.84%) vs. (Cr.13 =0%, Cr.18 =0%, Cr.21=0%, Cr.XY=0.33%), N.S]. Results were also evaluated according to the time elapsed between the PRE and POST -CT samples. No significant differences were found for seminal values (sperm concentration, p = 0.26; motility, p = 0.68), overall sperm DNA fragmentation (p = 0.68), double-strand DNA fragmentation (p = 0.90) and the frequency of sperm disomies (Cr. 13 p = 0.79, Cr.18 p = 0.46, Cr. 21 p = 0.85, Cr.XY p = 0.05). Limitations, reasons for caution The main limitations of the present study are that we only recruited patients who recovered spermatogenesis and that our data evaluated a long-term effect of CT. Wider implications of the findings All men wishing to have their own children after gonadotoxic treatment should receive adequate counseling about the side effects of the therapy. Based on our results on sperm DNA integrity and euploidy, patients who recovered spermatogenesis should not be advised to use samples that were preserved before undergoing CT. Trial registration number na

First results of the cultivation of prairie grass (Bromus catharticus Vahl) in Hungary

Yield elements (plant height, grass yield) of prairie grass, the little known grass species in Hungary, was investigated in a three-factor field experiment set up on the Mohács-island. One-grass and grass/legume mixture were established under irrigated and non-irrigated conditions at different nitrogen levels (0-67-133 kg N/ha/year). The effects of these factors on the grass productivity were analysed by a three-factor analysis of variance at 5% significance level in the first year of the experiment. Nitrogen supplementation and the interaction of legumeXirrigation had a significant positive effect on the annual hay yield. The yield of the first two cuts, which accounted for 61% of the annual yield, was not significantly affected by any of the examined factors, while their effect was already clearly visible in the yield of the 3rd and 4th cuts. A similar result was obtained with the model fitted to the average height of the four cuts, but the legumeXirrigation and legumeXnitrogen interactions were already significant in the 2nd cut. Nitrogen had a positive effect on grass height from the 3rd cut. Without irrigation, the 1st cut showed a significantly lower grass height with legume addition. In our experiment, we estimated 65 t/ha annual fresh grass yield on average of the treatments, and 18 t/ha of hay was actually harvested. In the treatment of irrigation with legume addition, 1 kg/ha of excess nitrogen resulted in an excess of 150 kg/ha of fresh grass yield in the case of the higher dose nitrogen treatment, compared to the nitrogen control. The laboratory analysis of the 1st cut-hay samples from selected treatments confirmed favorable protein (16.7-18.2%) and crude fat content, its lower crude fiber content and higher calcium and phosphorus content compared to cocksfoot and smooth brome.

THE RISK OF FOREST FRUITS CONTAMINATION BY HEAVY METALS IN THE HOREHRONIE REGION (SLOVAKIA)

The Horehronie region belongs to the oldest industrial areas in Slovakia. It is contaminated by residual acid tar dumps from times of Petrochema Dubová factory production since 1964. In the presented study, heavy metals (Cu, Cd, Ni, Zn, Pb) were determined in soils and forest fruits (Vaccinium myrtillus L., Vaccinium vitis-idaea L., Fragaria vesca L., Rosa canina L., Rubus idaeus L.) collected over two years from five locations in the Horehronie using Flame AAS method (Zn, Cu, Ni) and Graphite Furnace AAS method (Cd, Pb). The cadmium content, which was higher than its limit value in all analysed soils, poses the biggest threat. We determined more than 20 times the limit value for cadmium in the sample Veľká Smrekovica (9.17 mg/kg).The contamination factor (Cf) values and the geoaccumulation index (Igeo) indicate that this hazardous metal is involved in soil contamination to a large extent. The highest values of the degree of contamination (Cdeg) and the pollution load index (PLI) were determined in samples from the Ždiarska dolina and the Veľká Smrekovica. The contents of nickel and zinc were higher than the limit values only in the 1st sampling year in the Ždiarska dolina sample (46.30 mg/kg; 138.30 mg/kg). The results showed that soil monitoring and remediation of old environmental burdens are important in the territory of the Horehronie region. The highest permissible amounts for cadmium were exceeded in all types of forest fruits (0.03-0.18 mg/kg FW). The contents of nickel and lead were exceeded only in the sample of rosehips (Rosa canina). The contents of zinc and copper were lower than the permissible amounts in all samples. The estimated daily intake (EDI) was lower than the acceptable daily intake of all heavy metals. The bioaccumulation factor (BAF), total hazardous quotient and hazard index of the analysed samples was &lt; 1, which means that the consumption of forest fruits from the Horehronie does not pose a health risk for the consumer.

The genetics of autism and steroid-related traits in prenatal and postnatal life.

Autism likelihood is a largely heritable trait. Autism prevalence has a skewed sex ratio, with males being diagnosed more often than females. Steroid hormones play a mediating role in this, as indicated by studies of both prenatal biology and postnatal medical conditions in autistic men and women. It is currently unclear if the genetics of steroid regulation or production interact with the genetic liability for autism. To address this, two studies were conducted using publicly available datasets, which focused respectively on rare genetic variants linked to autism and neurodevelopmental conditions (study 1) and common genetic variants (study 2) for autism. In Study 1 an enrichment analysis was conducted, between autism-related genes (SFARI database) and genes that are differentially expressed (FDR<0.1) between male and female placentas, in 1st trimester chorionic villi samples of viable pregnancies (n=39). In Study 2 summary statistics of genome wide association studies (GWAS) were used to investigate the genetic correlation between autism and bioactive testosterone, estradiol and postnatal PlGF levels, as well as steroid-related conditions such as polycystic ovaries syndrome (PCOS), age of menarche, and androgenic alopecia. Genetic correlation was calculated based on LD Score regression and results were corrected for multiple testing with FDR. In Study 1, there was significant enrichment of X-linked autism genes in male-biased placental genes, independently of gene length (n=5 genes, p<0.001). In Study 2, common genetic variance associated with autism did not correlate to the genetics for the postnatal levels of testosterone, estradiol or PlGF, but was associated with the genotypes associated with early age of menarche in females (b=-0.109, FDR-q=0.004) and protection from androgenic alopecia for males (b=-0.135, FDR-q=0.007). The rare genetic variants associated with autism appear to interact with placental sex differences, while the common genetic variants associated with autism appear to be involved in the regulation of steroid-related traits. These lines of evidence indicate that the likelihood for autism is partly linked to factors mediating physiological sex differences throughout development.

INVESTIGATION OF GREEN SPACE COOLING POTENTIAL ON LAND SURFACE TEMPERATURE IN ANTALYA CITY OF TURKEY

Abstract. The aim of this study is to investigate the potential cooling effect of urban green spaces on land surface temperature (LST). Two significant green areas and their surroundings in the Konyaaltı district of Antalya province were selected as the sample area. Antalya is one of the most important tourist destinations in Turkey and has experienced rapid population growth along with increasing construction. Because the sample area is located in the Mediterranean Climate Zone, the research datasets were limited to a 6-month time series covering May and October, as recommended by previous studies. In the study, cloudless Landsat 8 satellite images were utilized to obtain vegetation density and land surface temperature. After making atmospheric corrections to the satellite images, the researchers used a series of algorithms to calculate NDVI and LST values from Landsat 8's NIR, Red, and thermal bands. A linear cross-section was assigned from the maps, and statistical analysis was performed by applying the Pearson correlation test to the data obtained from the pixels in contact with the section. The analysis revealed that there is a strong opposite correlation between plant density and land surface temperature, and the land surface temperature decreases as the plant density increases. The cooling effect of green areas range from 0.13–1.04 °C, although it varies according to the month. The 1st sample area had an average cooling effect of 0.77 °C, while the 2nd sample area had an average of 0.28 °C cooling effect. Combining the results from both sample areas, the researchers found that urban green spaces provide a cooling effect of 0.52 °C during the hot months of the spring and summer seasons in the Mediterranean climate zone. The results suggest that green areas play an important role in cooling today's cities that have turned into concrete jungles. Therefore, it is suggested that increasing the presence of urban green areas through green infrastructure planning can not only enhance the cooling effect of green spaces in urban areas but also help combat global climate change.

INFLUENCE OF FRESH BIOLOGICAL CHICKEN DROPPINGS ON THE MICROBIOLOGICAL INDICATORS OF LOW-PRODUCTIVE SODDY PODZOLIC SOIL

The article presents fresh biological dropping of caged chickens in crop production. After a thorough study of its environmental friendliness to the environment, this new biological dropping will reduce the cost of poultry farms for the disposal of dropping and use for more efficient cultivation of crops on marginal lands. The experiment used a high dose of organic matter - 120 t/ha, applied by spreading (control - without fertilisers). Sequencing of the metagenomes of soil samples showed that the soil microbiome’s phylogenetic composition was preserved due to the introduction of bacterial-fermented probiotic dropping.The authors also note that its taxonomic structure and functionality changed due to increased bacteria (with the different activity of taxa). Thus, the authors report an increase in the abundance of bacteria in the 1st sampling period from 6.9•107 to 1.3•108 copies/g of the soil of the 16S rRNA gene of bacteria in the plough horizon, and after 30 days - from 1.6•107 to 1.3×108 copies/g. For these purposes, opportunistic and pathogenic microorganisms in the fertilised soil found in all soil samples were also studied. A higher proportion of enterobacteria was noted in the control sample (0.6%), and with dropping, it did not exceed 0.15%. The content of pathogenic bacteria on a fertilised background in the 1st period of soil sampling practically did not change (0.7–0.8%), but in the 2nd period, it decreased sharply (from 2.5 to 1.1%). However, the authors note a decrease in the content of pathogenic bacteria. This decrease was accompanied by an increase in their abundance (from 3.9•105 to 1.4•106 copies/g), mainly Pseudomonas sp bacteria (from 2.1•105 to 9.3•105 copies/g), which is a consequence of a general increase in the abundance of bacteria in the microbiome against the background of enrichment of the soil with organic matter and an increase in soil fertility. The authors did not reveal a significant negative microbiological effect after the action of fresh biological chicken dropping on the soil’s ecological state at the specified research stage.

Influence of &lt;i&gt;Artemisiae absinthil herba&lt;/i&gt; and &lt;i&gt;Inulae rhizomata et radices&lt;/i&gt; on fermentation processes and methanogenesis in the rumen of young cattle

Relevance. Herbal preparations and their biologically active compounds can improve the use of feed and increase the productivity of animals by changing the microbial fermentation of the rumen. In our work, we evaluated the effect of herbal preparations — rhizomes and roots of elecampane and wormwood herb, on metabolic processes in the rumen and methanogenesis.Methods. For the study: Inulae rhizomata et radices were used in dosages per 1 kg of CB: 3.0 g — 1st sample, 1.0 g — 2nd sample, 6.0 g — 3rd sample; Artemisiae absinthil herba in dosages per 1 kg of CB: 5.0 g — 4th sample, 2.0 g — 5th sample, 10.0 g — 6th sample. The study was carried out in vitro using the ANKOM Daisy II incubator (modifications D200 and D200I) according to a specialized technique. The scar content was obtained from Kazakh white-headed bulls with chronic scar fistula with an average weight of 220–225 kg at the age of 9–10 months. Air sampling to determine the level of methane was carried out on the device «Kristallux-2000M» by gas chromatography. The level of volatile fatty acids (VFA) in the contents of the scar was determined by gas chromatography on a gas chromatograph «Crystallux-4000M», determination of nitrogen forms — according to GOST 26180-84.Results. It was found that different dosages of herbal preparations did not significantly affect the characteristics of fermentation in the gastrointestinal tract. Wormwood grass at a dosage of 10.0 g per 1 kg of SV reduced methane production than its other dosages (p ≤ 0.05). Thus, a dose of 2.0 g showed an increase in the level of methane in rye by 36.5% (p ≤ 0.01), a dose of 5.0 g — by 47.5% (p ≤ 0.01). Rhizomes and roots of elecampane showed a high level of methane production, however, min and max administered doses of this phytobiotic showed a decrease of 38.1–38.6%.