Commercial Kits Research Articles

Seroprevalence and associated risk factors for Fasciola hepatica in sheep in Nile Delta of Egypt

Fascioliasis is a globally distributed zoonotic parasitic disease that affects ruminants, including sheep. This study conducted from January to December 2023, aimed to determine the prevalence of Fasciola hepatica in sheep across three governorates in Egypt's Nile Delta and to assess associated risk factors. A total of 455 serum samples were analyzed using a commercial ELISA kit, revealing antibodies against F. hepatica in 22.2 % of the tested sheep. There was no significant association between locality or sex and the seroprevalence of F. hepatica in sheep; however, the highest prevalence was observed in Kafr Sheikh and in female sheep.Concerning risk factors, poor conditioned sheep aged between 1 and 2 years showed 2.1 and 3.8 times higher of infection probability than others. In addition, the risk of F. hepatica infection in sheep increased significantly in winter season (OR = 6.6, 95 %CI: 2.6–16.8), in absence of prophylactic treatment (OR = 2.2, 95 %CI: 1.3–3.3) and in presence of snail (OR = 3, 95 %CI: 3–5.4). The existence of antibodies against F. hepatica in examined sheep raising in Nile Delta indicating that the disease is reported in the studied areas and needs to be managed on farms through control and preventative measures.

Existencia/ausencia de los genes <i>stx2e</i> y <i>f18</i> en aislados de<i> Escherichia coli </i>de cerdos libres de diarrea

The aim of the study was to evaluate Escherichia coli isolates for the presence or absence of the f18 and stx2e genes. In total, 186 isolates obtained from piglets under 50 days of age that did not have diarrhoea at the time of sampling were studied. The animals were from four high-tech breeding farms located in the Lima region. Sampling was done in 2019 as part of a preliminary study using rectal swabs. A commercial purification kit was used for DNA extraction and a conventional PCR protocol was used for gene detection. No PCR product for stx2e or f18 could be detected in any of the E. coli isolated from these piglets.

Emerging zooanthroponotic risks: Detection of the human norovirus GII.4 Sydney[P31] strain in a domestic dog in Brazil

Recent increases in zoonotic diseases underscore the integration of companion animals into urban environments, posing complex transmission risks and highlighting the necessity of One Health approaches. Respiratory and enteric viruses have been consistently linked to interspecies transmission between humans and animals. This study aimed to assess the circulation of human noroviruses (NoV), human adenoviruses (HAdV), enteroviruses (EV), parechoviruses (PeV-A), human bocaviruses (HBoV), hepatitis A (HAV) and E viruses (HEV), Influenza A and B viruses (Flu A/B), respiratory syncytial virus (RSV), and SARS-CoV-2 in domestic dogs and cats in Brazil to understand potential zooanthroponosis risks. Between 2012 and 2021, 600 fecal samples from dogs and cats (516 and 84, respectively) were collected at small animal clinics in São Paulo state, Brazil. The specimens underwent in-house qPCR screening for HBoV and HAdV, while EV, PeV-A, NoV, and HEV were tested using in-house RT-qPCR. SARS-CoV-2, Flu A/B, and RSV were investigated with a commercial RT-qPCR kit assay. HAV detection utilized conventional nested (RT)-PCR. Positive samples were sequenced for molecular characterization and phylogenetic analysis. NoV was detected in 0.2 % (1/600) of the animals, while all other investigated viruses tested negative. The NoV-positive sample, collected in 2012 from a pet dog, was identified as genotype GII.4_Sydney[P31]. The Dog/BRA/2012/GII.4_Sydney[P31]/IAL-M21 strain exhibited a close genetic relationship to Brazilian human and environmental NoV GII.4_Sydney[P31] strains, with 98.1–99.2 % nucleotide similarity in ORF1 and 99.2–99.6 % in ORF2 sequences, suggesting interspecies transmission. Pet dogs are frequently exposed to human fecal-borne viruses, highlighting the potential for zooanthroponotic transmission due to their close interaction with humans in shared environments. There is an urgent need to enhance surveillance studies in companion animals to better understand the implications of detecting human NoV strains in pets, as NoV could potentially act as a reverse zoonotic disease in households, animal hospitals, or shelters worldwide.

Comparison of Three Different IBD Vaccination Protocols in Broiler Chicken-Efficacy, Serological Baselines and Histo-Pathological Lesions in the Bursa of Fabricius.

Infectious bursal disease (IBD) is a highly infectious disease of chicken found in all latitudes, due to the very high resistance to environmental conditions and commonly used disinfectants of the IBD virus (IBDV). This study was conducted to evaluate three IBD vaccination protocols in broiler chicken in terms of their effectiveness (clinical observations and production performance of the flock), estimation of serological baseline values (with the use of two different commercial ELISA kits) and the degree of progression of BF lesions (histopathological lesion score (HLS)) after vaccination. The three protocols were (I) single vaccination using an intermediate plus vaccine, (II) double vaccination with an intermediate vaccine and (III) double vaccination with an intermediate plus vaccine. Birds on farms vaccinated with protocol II were characterized by the lowest antibody titers in both ELISA tests and the lowest average HLS. The highest IDEXX titers were obtained in birds vaccinated with protocol III, while in the BIOCHEK test the highest titers were obtained for birds vaccinated with protocol I. Protocols I and III were characterized by similar HLSs. Birds vaccinated with protocols I and III had higher immune uniformity. The estimated serological baseline values and the degree of HLSs presents a clear picture of the differences between the different vaccination protocols and allows their adaptation for different farms depending on the current IBD epidemiological situation.

Genetic Relationships and Population Structure among Nigerian Ethnic Groups (Ibibio, Igbo, Hausa, Tiv and Yoruba) Based on Nine DNA Loci

Abstract Context: Population-specific attributes typically determine the behavior of human genotypes and traits in individuals. Examining the genetic relationships between populations presents an opportunity to observe patterns of genetic changes over time. By comparing populations, it is possible to see factors that may have sparked the evolution of a trait or genotype as well as the genetic diversity of the populations. Genetic diversity in the groups in Nigeria is still largely undefined. This can be achieved with the use of genetic markers. Aims: To determine the genetic relationships between Igbo, Ibibio, Yoruba, Tiv, and Hausa ethnic groups using nine short tandem repeat DNA markers. Settings and Design: Observational studies of 250 consenting participants. Methods and Material: Individuals of Igbo, Yoruba, Hausa, Ibibio, and Tiv were randomly sampled from their ethnic homes in Nigeria at participating institutions. DNA was extracted from blood samples using commercial DNA kits, and tested for nine autosomal short tandem repeat loci alleles using PCR and electrophoresis. The allele types and sizes were read and recorded for each locus for every individual. Statistical Analysis Used: Allelic frequencies, population pairwise genetic distances (FST and RST, analysis of molecular variance (AMOVA), and principal component analysis (PCA) were calculated using GenAlEx v6.502. Results: The fixation index (FST) was between 0.001 and 0.500 in the five populations. The estimated variation was 99.98% within individuals and 0.10% among populations. Four heterogeneous clusters were observed with three axes accounting for 32.86% variation. Conclusions: The genetic relationships closely mirrored linguistic groupings showing that language may have shaped early interaction and ultimately gene transfer between these groups.

CircSEC24A induces KLF8 expression to promote the malignant progression of non-small cell lung cancer by regulating miR-1253.

This study aimed to analyze the role of circSEC24A in non-small cell lung cancer (NSCLC) and its underlying mechanism. RNA levels of circSEC24A, microRNA-1253 (miR-1253), and KLF transcription factor 8 (KLF8) were detected by quantitative real-time polymerase chain reaction. Protein expression was analyzed by western blot or immunohistochemistry assay. Cell proliferation and apoptosis were investigated by colony formation assay, 5-ethynyl-2'-deoxyuridine assay, and flow cytometry analysis. Glycolysis was evaluated by commercial kits. Dual-luciferase reporter assay and RNA immunoprecipitation assay were conducted to identify the associations among circSEC24A, miR-1253, and KLF8. Xenograft mouse model assay was used to evaluate the effect of circSEC24A on tumor tumorigenesis. CircSEC24A and KLF8 were upregulated, while miR-1253 was downregulated in NSCLC. CircSEC24A knockdown inhibited proliferation and glycolysis but induced the apoptosis of NSCLC cells. CircSEC24A acted as a miR-1253 sponge and regulated NSCLC cell malignancy by targeting miR-1253. KLF8 was identified as a target of miR-1253, and its overexpression attenuated miR-1253-induced effects in NSCLC cells. Besides, circSEC24A upregulated KLF8 by sponging miR-1253. Further, circSEC24A knockdown suppressed NSCLC cell tumorigenesis in vivo. CircSEC24A silencing inhibited NSCLC cell malignancy through the miR-1253/KLF8 pathway, providing a potential therapeutic target for NSCLC.

Extraction of RNA from the fungus Moniliophthora Perniciosa (Stahel) Aime and Phillips-Mora cultivated in induction medium for characterization of chitininases

Chitin is considered the second most important biopolymer in nature, both for its physiological functions in different organisms and for its high availability in the biosphere. However, its natural accumulation does not occur in the environment, this demonstrates that there are natural mechanisms responsible for its degradation. The chitinolytic enzymes found in living beings are targets of great biotechnological interest because they degrade chitin into useful derivatives for various sectors of the economy. Given this, this study aimed to extract RNA for subsequent molecular characterization of chitinases from the fungus Moniliophthora perniciosa. To this end, the fungus was cultivated in WY medium, remaining in the greenhouse for 10-14 days. After growth, RNA was extracted using the traditional Trizol method and the Direct-zol commercial kit. The resulting samples were analyzed using the spectrophotometric technique and verified by photodocumentation after agarose electrophoresis. It was observed that the culture medium was considered satisfactory for RNA extraction and the ideal growth time was 8-10 days for the strain to present sufficient mycelial mass for extraction. Furthermore, the commercial kit used was more favorable than the traditional Trizol method, the product had a low level of contaminants and electrophoresis demonstrated high RNA quality due to the presence of intact rRNA bands. Therefore, as it was extracted from cultivation in a medium that induces chitinase production, the RNA extraction described becomes the most suitable method for the molecular characterization of chitinase production.

Evaluation of Label Compliance to Allergens in Imported Asian Food Sold in Northern Italy

Abstract Background The import of Asian food is strongly increasing in the EU. In Italy, the Asian market has expanded over the last 3 decades. Unfortunately, the labelling of these products often implies errors, mainly due to linguistic differences. Consequently, it can become a consistent problem regarding food safety, in particular for allergens declaration. According to EU Reg. 1169/2011 (Annex II), allergens must be labelled in bold and translated in the current language of the Member State. Asian food non-compliant labelling could enhance the probability of allergic consumers to get adverse events; accordingly, it is important to increase controls on the imported products. The present work investigates the label compliance to allergens on Asiatic products imported in Northern Italy. Methods Totally, 39 foodstuffs were tested for gluten, 93 for white egg proteins and 118 for milk proteins by commercial ELISA kits according to manufacturers’ instructions. Samples were considered non-compliant when the results were above 20 mg/Kg, the law limit for gluten, or above the milk LOD (10 µg/Kg) and the egg LOD (0.4 mg/Kg). Results Data showed 18% (CI 95% 0.09-0.32) of non-compliant samples for gluten: 6 samples > 80 mg/Kg (snacks and ready to eat meal) and 1 sample containing 48 mg/Kg (snack). In addition, 19.4 % (CI 95% 0.13-0.28) showed presence of egg proteins (noodles, snacks, patties and pasty). Finally, 13.6% (CI 95% 0.08-0.21) samples were irregular for undeclared milk (noodles and snacks). Conclusions Products label report their content and represent an important element for food trading and tracing. The results obtained confirm a high percentage of non-compliant labels, underlining how the consumption of imported products can represent a health risk for allergic consumers. The present work highlights the importance of imported food monitoring to guarantee the consumers health, and of the implementation of standardized/international regulated labelling rules. Key messages • Many non-compliant labels about allergens was observed on Asiatic products. • Correct labelling of allergens in imported food should be monitored to avoid adverse events.

Serum Malondialdehyde-Modified Low-Density Lipoprotein Level May Be a Biomarker Associated with Aortic Stiffness Among Patients Undergoing Peritoneal Dialysis

Background: Serum malondialdehyde-oxidized low-density lipoprotein (MDA-oxLDL) is associated with atherosclerosis and increased risk of cardiovascular disease (CVD). Vascular calcification frequently occurs with arterial stiffness in patients on peritoneal dialysis (PD). This cross-sectional study aimed to elucidate the correlation between aortic stiffness and MDA-oxLDL levels in patients on PD. Methods: Overall, 92 patients on PD were included. The carotid–femoral pulse wave velocity (cfPWV) was evaluated using cuff-based volumetric displacement, and blood samples were obtained from all patients. Aortic stiffness was classified based on cfPWV values (>10 m/s indicating aortic stiffness). Serum MDA-ox-LDL levels were quantified using commercial enzyme-linked immunosorbent assay kits. Results: In total, 33 (35.9%) patients were classified into the aortic stiffness group. Factors, including systolic blood pressure (SBP), serum triglyceride levels, C-reactive protein levels, age, weight, body mass index (BMI), waist circumference, MDA-oxLDL levels, and diabetes mellitus (DM) prevalence, were significantly higher in the aortic stiffness group. Multivariable logistic regression analysis revealed significant associations between aortic stiffness and MDA-oxLDL levels, BMI, and SBP. Furthermore, multivariable forward stepwise linear regression analysis revealed serum MDA-oxLDL levels as a significant independent predictor of cfPWV values. Conclusions: Serum MDA-oxLDL levels correlate positively with cfPWV values and may predict aortic stiffness among PD patients, highlighting its potential role in assessing CVD risk in this population.

Comparative evaluation of PCR and loop-mediated isothermal amplification (LAMP) assays for detecting Pasteurella multocida in poultry

ABSTRACT Aims To develop a colourimetric loop-mediated isothermal amplification (LAMP) assay for the detection of Pasteurella multocida in clinical poultry samples and compare the performance of this assay with PCR. A secondary aim was to evaluate a simple DNA extraction method that could enable LAMP-based testing in the field without the need for specialised laboratory equipment. Methods Primer sets for both LAMP and PCR were designed to amplify the KMT1 gene of P. multocida. DNA was extracted from 12 P. multocida isolates using a commercial extraction kit, and subjected to analysis using both LAMP and PCR. The analytical specificity of the LAMP assay was evaluated by testing it against a panel of 12 unrelated bacterial species, and the analytical sensitivity (limit of detection) was determined through testing of serial dilutions of the target DNA and compared to that of PCR. Subsequently, cloacal swabs (n = 40) from a commercial turkey flock were subjected to analysis using both LAMP and PCR assays, using a rapid DNA extraction method and a commercial extraction kit. Clinical sensitivity and specificity of the LAMP assay were calculated in comparison to PCR. Results A single DNA fragment of the expected size (∼ 200 base pairs), was amplified by PCR from 12 P. multocida isolates, which were also all positive by the LAMP assay. The identity of all PCR amplicons was confirmed by sequencing. Both PCR and LAMP showed similar analytical sensitivity, with a LOD of 20 pg of target DNA. As neither PCR nor LAMP assays produced positive results with 12 non-related bacterial species, the analytical specificity was assessed as 100%. However, LAMP demonstrated lower clinical specificity (94.74%) compared to PCR (100%) when 40 clinical samples were tested. None of the DNA samples extracted using the simplified DNA extraction method were amplified by either LAMP or PCR. Conclusion The LAMP assay developed in this study exhibits comparable performance to PCR in detecting P. multocida. Clinical relevance The use of a rapid and portable DNA extraction method, in conjunction with LAMP assays, could create opportunities for point-of-care testing for fowl cholera in field settings.

Evaluation of apoptosis markers in patients with atopic dermatitis

The purpose of the study was to analyze indicators of apoptosis in patients with atopic dermatitis (AD). Patients (N = 88) aged from 12 months to 3 years with varying degrees of disease severity were involved into trial. SCORAD indices were: 68.14±2.63 – in patients with severe AD; 32.03±1.43 – with moderate; and 12.12±1.43 – with mild AD, respectively. Determination of apoptosis markers (Caspase 8, Caspase 9, sCD153, sFas-L, Annexin 5) in blood serum was performed using an enzyme-linked immunosorbent assay (ELISA) using commercial kits from Bender MedSystems® (Austria). A study of apoptosis markers in the blood serum of children with AD revealed a significant decrease (p 0.05) in the concentration of Annexin 5 compared to standard values (by 20 times). A statistically significant decrease (p 0.05) in the concentration levels of Caspase 8 and Caspase 9 in the serum of AD patients was revealed. Thus, the concentrations of Caspase 8 were reduced on by 3 times, and Caspase 9 by 8 times compared to the norm. Also In addition, when determining sFas-L indicators, a statistically significant (p 0.05) decrease in concentration was noted compared to standard indicators. The content of sCD153 in the blood serum in AD patients was significantly higher (p 0.05) than the normative values by 5 times (not detected normally). The most pronounced disturbances in apoptosis indicators were observed in patients with severe atopic dermatitis, (p 0.05), highlighting the importance of studying apoptosis in the context of AD and the possible link between disruptions in this process and the severity of clinical manifestations in patients. The study demonstrates the peculiarities of apoptosis processes in children with AD. On the one hand, these patients have significantly increased activation of signaling systems, on the other hand, the elimination of altered immunocompetent cells does not occur properly, which contributes to the progression and chronicity of immune inflammation in the skin.

Comparison of the Characteristics of Circulating Small Extracellular Vesicles Isolated by Ultracentrifugation and a Commercial Kit.

The market offers a wide range of extracellular vesicles (EVs) isolation products, but their lack of standardization is a concern. Therefore, it is important to carefully assess the quality of the EVs obtained using these products. In this study, we compared the EXOCIB kit with the ultracentrifuge method, which is considered the gold standard for small EV isolation. After overnight fasting, small plasma EVs were extracted from four individuals using both the ultracentrifuge and the EXOCIB kit methods. The pooled EVs were then compared for the presence of the cluster of differentiation 63 (CD63) protein using the western blot analysis, and their size and zeta potential were performed by Dynamic Light Scattering (DLS). In addition, the size and morphology of small EVs were determined by using the Transmission Electron Microscopy (TEM) technique. An average hydrodynamic size of 135.7 nm and a zeta potential of -6.33 Mv at 25°C was found for small EVs isolated by the ultracentrifuge, whereas the kit method resulted in small EVs with a hydrodynamic size of 102.8 nm and a zeta potential of -0.907. Notably, the size of the particles in the kit samples was smaller compared to those obtained through the ultracentrifuge (P < 0.001). The western blot method confirmed the expression of CD63 in both methods, so the ultracentrifuge yielded small EVs with a higher level of purity compared to the kit-based approach (P = 0.036). The DLS findings revealed the existence of vesicles within the appropriate size range for small EVs like exosomes in both isolation techniques. The results of the western blot analysis, in conjunction with DLS, displayed that the ultracentrifuge method extracted small EVs with a greater degree of purity than the kit-based approach.

Effect of Particle Size on Physical Properties, Dissolution, In Vitro Antioxidant Activity, and In Vivo Hepatoprotective Properties of Tetrastigma hemsleyanum Diels et Gilg Powders

Objective: The aim of this study was to analyze the effects of different particle sizes of Tetrastigma hemsleyanum Diels et Gilg (TDG) powders on physical properties, dissolution, in vitro antioxidant activity, and in vivo hepatoprotective properties. Methods: The particle size of TDG coarse powders (TDG-CP), TDG fine powders (TDG-FP), and TDG micro powders (TDG-MP) were measured by a laser particle size analyzer. The physical properties were measured according to the latest version of the Chinese Pharmacopoeia (Committee Chinese Pharmacopoeia 2020). The content of the total flavonoids, total polysaccharides, kaempferol-3-O-rutinoside, and rutin of TDG powders were determined using the NaNO2-Al (NO3)3 colorimetric method, the sulphate-phenol colorimetric method, and HPLC, respectively. In vitro dissolution and antioxidant activity were determined by the paddle method in phosphate buffer (pH 6.8) and the DPPH radical scavenging method, respectively. In addition, the liver tissue pathology was evaluated by hematoxylin and eosin staining (H&amp;E), and the AST and ALT activities were measured by automatic biochemical analyzer. The superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) activities were measured by using commercial analysis kits. Results: As the particle size decreases, the fluidity of TDG powders decreased and the porosity increased. In addition, there were no significant differences in physical properties between low temperature pulverized powders and room temperature pulverized powders. The final dissolution rates of the four bioactive ingredients in TDG-MP were found to be 85.06%, 85.61%, 83.88%, and 83.26%, respectively, whereas in TDG-CP, the dissolution rates were significantly lower at 18.79%, 17.96%, 22.46%, and 24.35%. The EC50 values of TDG-CP, TDG-FP, and TDG-MP on DPPH scavenging activity were 0.82, 0.31, and 0.10 mg/mL, respectively. The AST and ALT activities of the TDG-FP group and the TDG-MP group were significantly decreased and the SOD, CAT, and GSH activities were significantly increased when compared with that of the model group. The inflammatory cell infiltration and vacuolar degeneration of liver cells in the TDG-FP group and the TDG-MP group were significantly improved. Conclusions: The particle size of TDG powders had a significant effect on the physical properties and in vivo bioactivity. TDG pulverized to a fine particle size or smaller is a promising approach for clinical applications with improved physicochemical and biological properties.

Genetic variability and discriminatory power of RM Y-STRs in gilgit men: a population study

AbstractShort Tandem Repeats (STRs) found on the Y chromosome have multiple applications, such as in forensic investigations, male identification for legal reasons, as well as population genetics. However, commercially accessible Y-STR tests are limited in their capability to distinguish closely linked male individuals in forensic genetics. Recent studies have revealed that Rapidly Mutating Y-STRs (RM Y-STRs) offer significantly greater haplotype diversity around the worldwide populations associated to conventional Y-STRs, though some RM Y-STR loci are not contained, current commercial kits. The objective of this research is to create a database of RM Y-STR haplotype frequencies for the Gilgit population in Pakistan and evaluate the effectiveness of these genetic markers in distinguishing individuals with genetic variations. The research involved examining several RM Y-STRs in 56 unrelated Gilgit men. The statistical analysis determined that the majority of the loci-maintained haplotype values, as specific varied in certain cases. The results presented that the RM Y-STRs were very effective in distinguishing genetic changes among the Gilgit population, through a GD value of 0 for DYD403S1 and GD values greater than 0.7 for all other loci, with maximum value of 0.9920 for the RM Y-STR DYS612.

TRP36-ELISA for E. canis detection: Concordance with TaqMan real-time PCR and point-of-care testing

Ehrlichia canis, a widely distributed tick-borne pathogen, requires prompt and accurate diagnosis for effective management. Real-time PCR serves as the reference standard for diagnosing E. canis infection, whereas serological tests are crucial for detecting antibody responses indicative of infection or post-exposure status. Early detection during the acute phase of the disease is essential for optimal treatment and recovery. E. canis Tandem Repeat Protein 36 (TRP36) elicits early acute phase responses. We developed an enzyme-linked immunosorbent assay (ELISA) using recombinant TRP36 to detect IgM and IgG antibodies against E. canis. We assessed the diagnostic agreement, sensitivity, and specificity of the rTRP36-ELISA and a commercial test kit (SNAP 4Dx Plus Test) with TaqMan Real-time PCR (qPCR) using the NCSS 2023 program version 23.0.1. Leftover serum samples from 32 dogs at the Veterinary Teaching Hospital were randomly selected and examined for E. canis infection using qPCR, rTRP36-ELISA, and SNAP 4Dx Plus Test. qPCR detected positive results in 12 (37.5 %), whereas ELISA and SNAP 4Dx Plus detected positive results in 10 (31.25 %) and 13 (40.62 %), respectively. Moderate agreement (κ = 0.545) was observed between rTRP36-ELISA and qPCR, and fair agreement (κ = 0.379) between SNAP 4Dx Plus and qPCR. Substantial agreement (κ = 0.79) was found between rTRP36-ELISA and SNAP 4Dx Plus. Compared to qPCR, the sensitivity and specificity of rTRP36-ELISA were 70 % and 77.27 %, respectively, compared to 53.85 % and 73.86 %, respectively, for SNAP 4Dx Plus. Our findings suggest that TRP36 is a promising antigen for E. canis serodiagnosis, potentially improving sensitivity and specificity.

Prognostic value of serum 25-hydroxyvitamin D levels and malnutrition status on postoperative complications in patients following laryngectomy with neck dissection.

Postoperative complications (PCs) following total laryngectomy remain a significant challenge, with recent investigations directed toward the impact of nutrition status and vitamin D deficiency. To elucidate the association between preoperative vitamin D level status, malnutrition risk score, and surgical and survival outcomes in patients with advanced laryngeal cancer following total laryngectomy. Prospective cohort study. Sixty-four patients with advanced laryngeal carcinoma treated with total laryngectomy were included in the study. Serum levels of 25(OH) D3 were measured employing a commercial chemiluminescent immunoassay kit, while nutrition status was evaluated using the nutrition risk index (NRI) and Malnutrition universal screening tool (MUST). The mean serum 25(OH) D level was 37.1 ± 19.4 nmol/L (range 11.0-100.6 nmol/L), with 47% of patients exhibiting vitamin D deficiency and 31% displaying insufficiency. Medium/high MUST score had 53% of patients, and moderate/severe NRI was verified in 48% of patients. Univariate logistic regression analysis identified MUST score, GPS score, neutrophil-to-lymphocyte ratio, and circulating 25(OH) D levels as predictive for the occurrence of PCs. In multivariate analysis, MUST score and circulating 25(OH) D levels remained significantly associated with PCs. Patients with high nutrition risk had significantly lower two-year OS rates compared to the medium and low nutrition risk groups, respectively (30% vs. 62% and 83%, p = 0.010). Early identification of malnourished or patients with vitamin D deficiency and those who would benefit from specific nutritional support could be beneficial for minimizing the risk of development of surgical complications and help improve our clinical outcomes.

Emodin-based Regulation and Control of Serum Complement C5a, Oxidative Stress, and Inflammatory Responses in Rats with Urosepsis via AMPK/SIRT1

Emodin, derived from Rheum officinale and aloe, is known for its diverse benefits such as anti-inflammatory, antioxidant, and antibacterial properties. Currently, the impact of emodin on urosepsis is unclear. This study aims to investigate the mechanism of action of emodin in urosepsis. Peripheral blood mononuclear cells (PBMCs) were purchased from Cloud-Clone Animal Inc. and treated with emodin. Cell viability and the lactate dehydrogenase (LDH) level were then assessed. In a separate experiment, a urosepsis model was established in Sprague Dawley rats which were subsequently treated with emodin. The levels of oxidative stress-related factors, serum complements and inflammatory factors were measured using commercial kits. Blood urea nitrogen and serum creatinine levels were determined using a fully automatic biochemical analyzer. The levels of pro-inflammatory proteins and AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway-related proteins were evaluated via Western blot. PBMCs were unaffected by emodin concentrations below 60 μg/mL, and minimal LDH levels were detected in the cells. Emodin attenuated the effects of Escherichia coli and diminished the production of serum complements, oxidative stress-related proteins, and inflammatory factors in PBMCs. Notably, the effects of emodin were lessened by an AMPK pathway inhibitor. Additionally, emodin alleviated oxidative stress, complement system activation, inflammation, and kidney injury in urosepsis rats through the AMPK/SIRT1 signaling pathway. Emodin improved kidney damage in urosepsis rats by activating the AMPK/SIRT1 signaling pathway, which reduced oxidative stress, inflammation, and complement system activation.

Association of HPA Antigens with Immune Thrombocytopenia: A Case-Control Study by PCR-SSP Method

Background: Human platelet antigens (HPAs) play a clinically significant role in alloimmunization and the development of immune-mediated disorders such as immune thrombocytopenia (ITP), fetal and neonatal alloimmune thrombocytopenia (FNAIT), and post-transfusion purpura (PTP). Understanding the genetic profiles of HPAs is critical for preventing and treating these conditions. Given the limitations of serological methods in determining HPA genotypes, this study aims to investigate the association between the genotypes of HPA1, HPA2, HPA3, HPA4, and HPA15 antigens and autoimmune thrombocytopenia in Lorestan Province, utilizing the PCR-SSP method. Materials and Methods: This case-control study involved 80 individuals diagnosed with ITP and 120 healthy controls. DNA samples were extracted using a commercial DNA extraction kit, with concentrations quantified via a Nanodrop spectrophotometer. Genotyping was performed using the PCR-SSP method with specific primers for each HPA gene. The genotype data were verified using previously established sample sets. The frequencies of each HPA genotype were recorded, and a comparative analysis was conducted between the patient and control groups to evaluate the study hypothesis. Results: The results revealed that individuals carrying the HPA2b allele had a 5.31-fold increased risk of developing ITP, a statistically significant finding (P &lt; 0.05, OR = 5.31). Similarly, the presence of the HPA15b allele was associated with a 6.54-fold increased risk (P &lt; 0.05, OR = 6.54). Conclusion: These findings, in conjunction with previous studies, suggest the need for larger-scale investigations across different populations. Such research could aid in the early diagnosis and prediction of thrombocytopenia severity, inform treatment strategies, and facilitate the removal of pathogenic antibodies from circulation.

β-Sitosterol Mitigates Apoptosis, Oxidative Stress and Inflammatory Response by Inactivating TLR4/NF-кB Pathway in Cell Models of Diabetic Nephropathy.

Podocyte injury plays a pivotal role in the pathogenesis of diabetic nephropathy (DN), leading to proteinuria formation. β-Sitosterol is a natural compound with anti-inflammatory, anti-diabetic, nephroprotective and antioxidant properties. The studyaimed to explore whether and how β-Sitosterol protected podocytes against high glucose (HG)-induced inflammatory andoxidative injury. DN cell models were established by stimulating podocytes or renal tubular epithelial cells (HK-2) cells with 25 mM glucose. Cell viability and apoptosis were evaluated using cell counting kit-8 assays and flow cytometry analyses. Westernblotting was used to quantify protein levels of genes related to podocyte injury, HK-2 cell damage, inflammation, and TLR4/NF-кB pathway. Contents of oxidative stress biomarkers were evaluated by corresponding commercial kits while proinflammatorycytokine levels were determined by enzyme-linked immunosorbent assay. Immunofluorescence staining was performed todetect intracellular levels of reactive oxygen species (ROS) and Nrf2 nuclear translocation. Experimental results revealed that HG treatment induced podocyte dysfunction by impairing cell viability while accelerating theapoptosis, and the changes were reversed by β-sitosterol treatment. Moreover, β-sitosterol repressed HG-evoked oxidative stressby reducing ROS and malondialdehyde (MDA) levels while increasing activities of antioxidant enzymes. The reduction ofproinflammatory cytokines mediated by β-sitosterol in HG-stimulated podocytes suggested the anti-inflammatory role of β-sitosterol. Additionally, the activation of the TLR4/NF-кB signaling induced by HG was inhibited by β-sitosterol in podocytes.Inactivation of the TLR4 using TAK-242 enhanced the protective effects of β-sitosterol against HG-mediated oxidative stressand inflammation. Similarly, β-sitosterol also protected HK-2 cells from HG-induced oxidative stress, inflammation, andapoptosis. In summary, β-sitosterol exerts anti-inflammatory, anti-oxidative, and anti-apoptotic activities in HG-induced podocytes or HK-2 cells by inhibiting TLR4/NF-кB signaling.

Transplacental Transfer of Oxytocin and Its Impact on Neonatal Cord Blood and In Vitro Retinal Cell Activity.

The development of fetal organs can be impacted by systemic changes in maternal circulation, with the placenta playing a pivotal role in maintaining pregnancy homeostasis and nutrient exchange. In clinical obstetrics, oxytocin (OXT) is commonly used to induce labor. To explore the potential role of OXT in the placental homeostasis of OXT, we compared OXT levels in neonatal cord blood among neonates (23-42 weeks gestation) whose mothers either received prenatal OXT or experienced spontaneous labor. Our previous research revealed that the oxytocin receptor (OXTR), essential in forming the blood-retina barrier, is expressed in the retinal pigment epithelium (RPE). We hypothesized that perinatal OXT administration might influence the development of the neural retina and its vasculature, offering therapeutic potential for retinal diseases such as retinopathy of prematurity (ROP). Plasma OXT levels were measured using a commercial OXT ELISA kit. Human fetal RPE (hfRPE) cells treated with OXT (10 µM) were assessed for gene expression via RNA sequencing, revealing 14 downregulated and 32 upregulated genes. To validate these differentially expressed genes (DEGs), hfRPE cells were exposed to OXT (0.01, 0.1, 1, or 10 µM) for 12 h, followed by RNA analysis via real-time PCR. Functional, enrichment, and network analyses (Gene Ontology term, FunRich, Cytoscape) were performed to predict the affected pathways. This translational study suggests that OXT likely crosses the placenta, altering fetal OXT concentrations. RNA sequencing identified 46 DEGs involved in vital metabolic and signaling pathways and critical cellular components. Our results indicate that the perinatal administration of OXT may affect neural retina and retinal vessel development, making OXT a potential therapeutic option for developmental eye diseases, including ROP.