GraphPad Software Research Articles

Molecular genetic characteristic of pulmonary tuberculosis associated with ABCB1 gene expression of multidrugresistance protein P-gp

Background: Tuberculous inflammation is mediated by a complex molecular signaling pathway, the analysis of which makes it possible to identify promising biomarkers and targets for the development of new diagnostic, prognostic and pharmacological approaches in order to improve the effectiveness of anti-tuberculosis chemotherapy. Determining the relationship between key inflammatory cytokines, the multidrug-resistant protein P-gp and the activity of specific inflammation in the surgical material of patients with pulmonary tuberculosis may prove to be a novel tool in the development of pathogenetic therapy and personalized medicine. Aims: to characterize molecular and genetic profiles of tuberculomas and identify genes that correlate with the expression of the ABCB1 gene of the P-gp protein in the surgical material of patients with pulmonary tuberculosis. Research objectives: 1) to obtain molecular and genetic characteristics of tuberculosis by real-time PCR and compare it with the activity of tuberculous inflammation; 2) to carry out a correlation analysis between the expression of the ABCB1 gene and key cytokines of the tuberculosis process: IL-6, IL-10, IFN-γ, TGF-β, TNF-α, IL-1β. Materials and methods: A prospective cohort study was conducted on the basis of the FSBI CTRI. The object of the study was the surgical material of 35 patients diagnosed with multiple pulmonary tuberculomas. Histological examination methods were used for the morphological assessment of the surgical material. A real-time quantitative PCR method was used to analyze gene expression. Statistical processing was performed using the GraphPad Prism Version 7.04 software package (GraphPad Software, USA). The data is presented as a median with an interquartile range. The nonparametric Mann-Whitney U-test was used to compare the two groups. All p-values were two-sided and p 0.05 was considered statistically significant. The correlation between the variables was estimated using the Spearman correlation coefficient. The correlation analysis was carried out in the Microsoft Office Excel 2010 software. Results: The study revealed that the highest level of expression of ABCB1 gene of the P-gp protein is observed in tuberculomas with high activity of tuberculous inflammation, and its expression is correlated with the expression of the IL6 gene (p0.001) and the expression of the IL10 gene (p0.01). Tuberculomas of this group are also characterized by higher expression of the TGFB1, TNF and IL1B genes, compared with the group of moderate activity of specific inflammation. Conclusions: The data obtained indicate that in addition to pro-/anti-inflammatory cytokines, the P-gp protein plays an important role in the pathogenesis of tuberculous inflammation, especially with its high activity. Further clarification of the P-gp role in tuberculous inflammation may be an important step for the development of new approaches to treat tuberculosis using methods of HDT and personalized medicine.

The use of new psychoactive substances among young adults in Romania

Background and objectives. The consumption of new psychoactive substances (NPS), particularly among young people, remains a significant public health issue. The principal objective of this study was to identify the incidence and prevalence of NPS use among young people in Romania and the impact of the consumption on their mental health. Materials and methods. A cross-sectional study was conducted on a sample of 231 Romanian participants on March-April 2023. Data were collected through an online questionnaire and analyzed using GraphPad and Excel statistical software. Results. The prevalence of NPS use was 38.10% within the sample The analyzed data indicated an increased incidence of NPS use among male participants. Notably, most respondents who reported NPS use were students, accounting for 23.38% of users. Among the total of 88 NPS users, 22.68% consumed synthetic cannabinoids, 14.09% bath salts, and 13.75% used mephedrone. Less frequently consumed substances included synthetic opioids, methamphetamines, synthetic piperazines, Salvia divinorum, and MDPV. A significant proportion of users reported severe psychiatric symptoms including psychosis, depression and anxiety. Among non-users, awareness of the side effects of NPS influenced over half of them to avoid such substances. Conclusions. NPS use impacts predominantly young people. Organizing prevention and information campaigns about the implications of NPS use and its immediate harmful effects could help reduce its prevalence among young individuals.

Leptin/Melanocortin Pathway in Cholelithiasis Patients: A Diagnostic Perspective?

Cholelithiasis is the most prevalent inflammatory condition of the gallbladder. The regulation of biological processes, including energy homeostasis, and control of body weight are key mechanisms that the leptin and melanocortin pathways play a role in. Cholelithiasis is the most prevalent inflammatory condition of the gallbladder. There are various risk factors for the development of gallstone disease, especially weight gain, and obesity is just one of them. This risk factor can be minimized by maintaining appetite and energy balance. Here, leptin and melanocortin pathways are the key mechanisms in maintaining appetite and energy homeostasis. The aim of this study was to investigate the relationship between the levels of LEP, LEPR, TrkB, BDNF, POMC, and MC4R proteins in patients with Cholelithiasis. This study aims to determine the relationship between LEP, LEPR, TrkB, BDNF, POMC, and MC4P protein levels, which play a role in maintaining appetite and energy homeostasis, and cholelithiasis. This study examined 44 patients diagnosed with Cholelithiasis and 44 healthy control subjects who had not previously been diagnosed with any form of Cholelithiasis. The levels of leptin (LEP), Leptin Binds To Leptin Receptors (LEPR), Tropomyosin Receptor Kinase B (TrkB), Brain-Derived Neurotrophic Factor (BDNF), Pro-OpioMelanoCortin (POMC), and Melanocortin- 4 Receptors (MC4R) molecules were analyzed using the Enzyme-Linked Immunosorbent Assay (ELISA) method. The results were analyzed using the SPSS Software (Version 22.0) program and GraphPad Prism 8.0.1 software. The study found a statistically significant decrease (p < 0.05) in MC4R, TrkB, BDNF, and POMC protein levels in Cholelithiasis patients compared to the control group. There was no statistically significant difference in LEP and LEPR concentration values between the two groups (p=0.247, p=0.674). The proteins MC4R, TrkB, BDNF, and POMC, which are involved in the leptin and melanocortin pathways may play a significant role in Cholelithiasis disease. However, more detailed research on the relevant proteins is needed. Nevertheless, this research will guide new studies.

Pengaruh Penambahan Ekstrak Bunga Sepatu (Hibiscus rosa sinensis Linn.) Terhadap Peningkatan Aktivitas Antioksidan, Total Flavonoid, dan Warna Pada Mie Kering Berbahan Mocaf

Dry noodles are a popular type of noodle, but product innovation needs to be carried out to promote food diversity. Hibiscus flower extract can be used not only as a natural coloring agent but also for its health benefits, such as antioxidant activity and total flavonoids. The aim of the research was to identify the effect of adding hibiscus extract to dry noodles on antioxidant activity, total flavonoids, moisture content and color. A non-factorial Completely Randomized Design with five treatments and three replications was used. The ratio of hibiscus extract was 5, 10, 15, 20 and 25% with a wheat flour and mocaf ratiom of 60:40. Mocaf was used in this product because it is affordable, widely available, and gluten free. The result of ANOVA (5% level) and Tukey’s test using Graphpad software showed that the addition of hibiscus extract to dry noodles had a significant effect on all test. The antioxidant activity in the best treatment, with 25% hibiscus extract was 47.482%, which was classified as very strong. The total flavonoids in the best treatment with 25% hibiscus extract was 147.14 mgQE/g, The mositure content ranged between 2.95-7.68% (meeting standards of SNI 01-8217-2015). The color of the dry noodles ranged from 0.34-1.44, with differences classified from not clearly visible to very small.

Investigating the effect of Iaraj Fiqra capsule and dry cupping on knee osteoarthritis: A clinical trial study

Background: Knee arthritis is a joint disease caused by the destruction of the underlying cartilage and bone. Common symptoms of this disease are stiffness and pain. Dry cupping and Iaraj Fiqra capsules are used in Persian medicine to treat knee arthritis. Therefore, the present study aimed to assess the effect of the Iaraj Fiqra capsule and dry cupping on arthritis and compare their effects. Materials and Methods: This randomized, single-blind trial was conducted on 61 subjects aged 60 to 69 years old with knee osteoarthritis after obtaining informed consent. Patients were randomly assigned to two groups. One group received 500 mg/day of Iaraj Fiqra capsules for four weeks (n=31), and the other group received dry cupping on the knee for 10 min twice a week for four weeks (n=30). Patients completed the Knee Injury and Osteoarthritis Outcome Score questionnaire, which assessed five areas: pain, symptoms, daily activities, sports and recreation function, and knee-related quality of life, both before and after the study. Independent and paired t-tests, Fisher's exact test, and the Chi-square test were used to evaluate the data in GraphPad software (version 8). Results: The results demonstrated that Iaraj Fiqra capsules and dry cupping for four weeks improved pain (Iaraj Fiqra capsule: before: 92.1±1.3 and after: 73.1±1.4, p=0.002, dry cupping: before: 96.6±2.3 and after: 83.0±3.1; P=0.02), symptoms (Iaraj Fiqra capsule: before: 84.12±3.1 and after: 73.23±1.5; P=0.005, dry cupping: before: 82.11±1.2 and after: 67.31±1.1; P=0.001), daily activities (Iaraj Fiqra capsule: before: 32.22±2.5 and after: 76.21±0.6; P=0.002, dry cupping: before: 34.15±1.4 and after: 72.12±1.2; P=0.001), sports performance (Iaraj Fiqra capsule: before: 92.1±1.3 and after: 73.1±1.4; P=0.002, dry cupping: before: 96.6±2.3 and after: 83.0±3.1; P=0.001,) and recreational (Iaraj Fiqra capsule: before: 24.21±1.2 and after: 68.11±0.5; P=0.002, dry cupping: before: 28.95±1.2 and after: 62.25±2.7; P=0.001), and knee-related quality of life scores (Iaraj Fiqra capsule: before: 31.11±0.6 and after: 82.32±11.4; P=0.002, dry cupping: before: 11.6±33.11 and after: 81.21±1.4; P=0.002). Furthermore, the results demonstrated that the pain score of patients improved significantly compared to the dry cupping group (P&lt;0.05). Conclusion: As evidenced by the obtained results, Iaraj Fiqra capsules and dry cupping can be used to improve the pain, symptoms, daily activities, sports and recreation function, and knee-related quality of life of individuals with knee osteoarthritis.

SINGLE INSTITUTION RETROSPECTIVE ANALYSIS OF COMPLICATIONS ASSOCIATED WITH SURGICAL DEBULKING OF BRAIN METASTASIS

Abstract AIMS Surgical resection of brain metastasis is associated with complications, which worsen patient’s quality of life, results in oncological treatment delays and death. Surgical complication data aids decision making, leads to objective risk benefit analysis and informed consent. The aim of this study is to quantify rates, types and risk factors for complications related to surgical debulking of brain metastasis. METHOD Retrospective analysis on adult patients with brain metastasis, who underwent surgical debulking between 2017 and 2023 at Royal Preston Hospital. Data collected included demographics, diagnosis, 30-days post-operative complications, 30-days mortality, 30-days post-operative palliation, primary tumour, Karnofsky-Performance Score (KPS), first vs. repeat resection, use of drain and type of suture materials etc. Univariate analysis was carried out using Graphpad software. RESULTS Two-hundred and twenty-seven patients qualified. The most common primaries were lung (n=93, 41%), breast (n=43, 18.9%), and skin (n=31, 13.7%). The most common complications were Hospital-acquired pneumonia (HAP) (n=27, 11.9%), surgical site infection (SSI) (n=19, 8.4%), wound dehiscence (n=14, 6.2%), thromboembolic events (n=12, 5.3%) and CSF leak (n=8, 3.5%). Thirty days mortality and post-operative palliation (due to surgical complications) were both 3.5% each (n=8). Two-hundred and six (93.2%) had KPS ³70 whilst 15 (6.8%) had KPS &amp;lt; 70 [range 30-60 pre-operatively (median 60)). KPS &amp;lt; 70 was statistically significant for post-operative palliation (p=0.01), HAP (0=0.02) and urine infection (p=0.002), but not for 30-day mortality. Twenty-five (11.1%) patients undergoing repeat resections, experienced higher wound infections (p=0.002). CONCLUSION Surgical debulking is associated with significant post-operative complications, with 7% either dying or not receiving any further treatment, 18.1% percent developing serious wound complications (SSI, CSF leak, Wound dehiscence) and 19.4% experiencing significant medical complications (HAP/UTI/Thromboembolism). Pre- operative KPS &amp;lt; 70 and repeat resections are associated with statistically increased risk of complications.

Biomarker Profiles and Clinicopathological Features in Head and Neck Squamous Cell Carcinoma Patients.

Background and Objectives: Head and neck squamous cell carcinomas (HNSCCs) vary significantly in terms of invasiveness, growth rate, and metastatic potential. This study aimed to investigate the expression of several prognostic biomarkers (Ki67, p53, EGFR, COX-2, Cx43, and p16) in HNSCC from various anatomical regions and to correlate these expressions with clinicopathological parameters. Materials and Methods: We performed immunohistochemistry on 91 histologically verified HNSCC cases from the County Emergency Hospital, Targu Mures. Biomarker expression for Ki67, COX-2, and Cx43 was assessed using a standard immunoexpression scoring system: S1: 0-10%, S2: 11-25%, S3: 26-50%, S4 > 50%; EGFR was scored based on membrane staining intensity: 0, 1+, 2+, 3+; we classified p16 as positive or negative; p53 was grouped into mutant and wild-type; and we compared these across histopathological types, tumor grades, anatomical locations, gender, and different age groups. We performed a comparative analysis of Cx43 expression levels in relation to the expression of the rest of the markers. Statistical analysis was conducted using GraphPad InStat 3 software, version 3.06 (GraphPad Software Inc., San Diego, USA). Results: The majority of tumors were in males (95.6%) aged 51-60 years. Mutant p53 expression was prevalent in most cases. Elevated Ki67 and EGFR expression were associated with more aggressive tumors. COX-2 levels varied, with a higher proportion of moderate and high immunoexpression (S3 + S4) observed in patients under 70 years old. Cx43 expression was generally low, especially in extralaryngeal tumors. Conclusions: HNSCC primarily affects older males, with the larynx being the most common site. High levels of Ki-67 and EGFR suggest more aggressive tumors, while low COX-2 levels reflect varying prognoses. Women may develop more aggressive tumors, and extralaryngeal tumors often present with more challenging prognoses. Low Cx43 expression may be more likely to coincide with higher Ki67 and COX-2 levels, possibly indicating a link with more aggressive tumor behavior.

A bibliometric analysis and visualization of literature on the relationship between vitamin D and obesity over the last two decades

ObjectiveThe purpose of this study was to employ bibliometric analysis to visualize hot spots and evolving trends in the studies on the relationship between vitamin D and obesity. MethodsFrom the Web of Science Core Collection database, articles on vitamin D and obesity from 2001 to 2021 were retrieved. For the bibliometric visualization analysis, CiteSpace was employed. Some of the figures were created using GraphPad software. Results4454 pieces of articles and reviews were found, with an average citation of 30.68 times. There are many more published papers in the area of "nutrition dietetics" (1166, 26.179 %). The United States possesses the largest number of publications (1297, 29.12 %) and demonstrates definitive leadership in this field. The League of European Research Universities generates a higher percentage of publications (256, 5.748 %) than other institutions. Major studies are funded by the United States Department of Health and Human Services (531, 11.922 %) and the National Institutes of Health, USA (528, 11.855 %). The top five keywords with the highest co-occurrence frequency are “obesity” (1260), “vitamin d” (943), “insulin resistance” (651), “risk” (642), and “d deficiency” (636). The biggest keyword cluster was #0 "adolescent" among the 18 keyword clusters. The three latest keywords in the keyword burst were "mineral density"、"d insufficiency" and "25 hydroxyvitamin d concentration". ConclusionThis bibliometric analysis shows an overview of the current status of the research on the association between vitamin D and obesity. The prevalence of vitamin D deficiency and the relationship between vitamin D and metabolic syndrome in obese individuals remains hot topics. We speculate that the effect of obesity on vitamin D levels and bone mineral density, and the influence of vitamin D insufficiency on various body systems in obese populations will be future trends.

Camel FGF21 Protein Enhances FGF21 Signaling and its Targeted Compounds Screening.

The hormone Fibroblast Growth Factor 21 (FGF21), as a novel glucose and lipid metabolism regulator, has become a promising therapeutic target for metabolic disorders. Camel, the characteristic species adapted to arid and semi-arid desert climates, has shown exceptional ability for the regulation of lipid reserve and utilization. This study found camel FGF21 to have a stronger regulatory effect for downstream signaling compared to human and mouse through sequence analysis and FGF21 signal downstream markers detection. FGF21 protein has been found to have one potential drug-binding pocket, which has been predicted using the CavityPlus online platform. Four small compounds, resorcinol monoacetate, tropisetron, nylidrin, and stiripentol, targeting FGF21 protein have been screened by molecular docking using the UCSF DOCK6 program. The inhibitory concentration 50% (IC50) values of the four small compounds have been determined by MTT assay and the values have been simulated by the software GraphPad Prism. The biological effect testing has indicated the four compounds to be involved in the regulation of the FGF21 signaling pathway and serve as agonists for FGF21 signaling transduction. While the blocking experiment of compound and protein cotreatment has indicated the four small compounds to not inhibit FGF21-induced pathway activation. Even, resorcinol monoacetate and stiripentol have shown to synergistically activate downstream signaling pathways with the FGF21 protein. This study has provided new ideas for developing therapeutic strategies based on FGF21 protein modification and exploring novel disease treatment strategies based on the compounds-protein combination.

MRNA Relative Expression Catalase in Hypertension

Hypertension is number one worldwide disease which lead to death. Initially hypertension is caused by changes in the walls of blood vessels. These vascular changes are due to the accumulation of ROS (Reactive Oxygen Species). ROS is the result of the balance between antioxidants and oxidants, but uncontrolled conditions will cause vascular damage. One of ROS product from metabolism which could played role in downregulation antioxidant gene expression is hydrogen peroxide. Hydrogen peroxide could be neutralized by catalase enzyme. Studies about catalase expression in hypertension subject is still limited. This research aims to determine mRNA relative expression catalase in hypertension. In this case control study, thrity subject hypertension and normotension subject were recruited within the age of 50-60 years. Hypertension subject was chosen according to JNC 8. Two millilitres vein blood was isolated into RNA. mRNA expression was detected by qRT PCR with 2 steps. Relative expression was determined using livak method. The data was analyzed using Mann Withney test with GraphPad software. Research results show that (1) catalase mRNA relative expression was 0,6 fold in hypertension; (2) catalase relative expression was not significantly lower in hypertension; (3) other antioxidants might have other mechanism to detoxify ROS and prevent hypertension.

Children With Acute Lymphoblastic Leukemia in Romania: Results From a Decade-Long Single-Center Study.

Acute lymphoblastic leukemia (ALL) is the most prevalent cancer in children, with continuously improving survival rates. As few studies in Romania have analyzed ALL patients and disease characteristics or survival, we conducted a retrospective study on 158 patients diagnosed with ALL admitted to the Department of Pediatric Oncology and Hematology at the Emergency Hospital for Children, Cluj-Napoca, Romania, from January 2011 until April 2021. The most important objectives of the study are to establish full profiles of the patients and ALL, remission rates, relapses, and deaths, an epidemiology analysis to determine the incidence of ALL for comparison with the standard European population, and also to assess survival by the most important parameters, including minimal residual disease (MRD). This was a retrospective study that focused on patients with newly diagnosed ALL from January 2011 to April 2021 in the Department of Pediatric Hematology and Oncology of the Emergency Hospital for Children, Cluj-Napoca, Romania. The inclusion criteria were: patients with de novo ALL, patients who were younger than 18 years of age, and patients with signed informed consents. The exclusion criteria included patients who were older than 18 years of age; patients with relapsed ALL; and patients who did not have a signed informed consent. A total of 158 patients were included in the study, aged between 0 and 17 years. The information about patient characteristics and all variables was taken from the patients' files, where informed consent is mandatorily stored. Descriptive statistics were used to evaluate variables such as age, gender, leukocyte number, immunophenotype, prednisone response, risk group assessment, cytogenetics, relapse, and MRD. Kaplan-Meier curves were used to assess survival, for which patients were divided into two subcohorts: a 2011-2015 subcohort and a 2016-2020 subcohort. The statistical analyses used Prism version 10.2.3 (GraphPad Software, La Jolla, CA). For the analysis, we used Kaplan-Meier curves, the log-rank (Mantel-Cox) test, the log-rank test for trends, the Gehan-Breslow-Wilcoxon test, survival proportions, the chi-square test, and correlations. A shift in risk groups was observed after the introduction of MRD testing in 2017, with more patients being stratified in the medium-risk group (MRG) and high-risk group (HRG). At the survival analyses between bone marrow (BM) aspiration and MRD on day 15, we discovered that patients with MRD>10% had much higher overall survival (OS) and event-free survival (EFS) compared to patients with >25% blasts in the BM; 24(11.4%) patients relapsed, of which, nine (3.8%) were very early relapses and 10 (4.1%) were late relapses. The five-year OS and EFS for patients with the T-cell immunophenotype of ALL and those with leukocyte counts >100,000/mm3were identical to those at one year. The OS of the 2016-2020 subcohort was higher than that of the 2011-2015 subcohort, and more patients were stratified into MRG and HRGs due to the implementation of MRD testing. Minimal residual disease testing helped to improve significantly the survival rates of patients with more than 10% residual disease. None of the patients with very early relapses entered complete remission (CR), but all late relapses achieved CR. All events experienced by patients with the T-cell immunophenotype, or leukocyte counts of >100,000/mm3 occurred in the first year after diagnosis.

Evaluating the Efficacy of Actinidia deliciosa (Kiwi Fruit) Extract in Inhibiting Pseudomonas aeruginosa Biofilm Formation: An In Vitro Study With Therapeutic Implications.

Background Gram-negative Pseudomonas aeruginosa is a common bacteria that is well-known for its capacity to build biofilms, which are organized cell communities encased in a self-produced polymeric matrix. Treating infections becomes more challenging due to biofilms' capacity to provide immunity and resistance to antibiotics. The search for novel anti-biofilm agents has gained significant momentum, and the diverse range of bioactive compounds found in natural products offers a promising avenue. Rich in vitamins, antioxidants, and various phytochemicals, Actinidia deliciosa (kiwi fruit) has demonstrated potential as an antibacterial agent. Aim of the study This study aims to assess the efficacy of A. deliciosa extract in inhibiting biofilm formation by P. aeruginosa in vitro, providing valuable insights into its potential as a natural therapeutic agent for preventing recurrent bacterial infections. Materials and methodology The antibacterial and antibiofilm properties of A. deliciosa (kiwi fruit) methanolic extract were assessed in this study against P. aeruginosa (PAO1). The fruit was gathered, examined by a botanist for authenticity, and then cold macerated in methanol to create an extract. A two-fold broth dilution procedure was used to calculate the minimum inhibitory concentration (MIC), and agar well-diffusion was used to evaluate the antimicrobial activity. Pyocyanin pigment quantification was carried out after the extract was applied, and the antibiofilm impact was evaluated using a crystal violet assay. GraphPad Prism (GraphPad Software, San Diego, CA) was used for statistical analysis. Results Based on our findings, A. deliciosa was shown to have significant antibacterial and antibiofilm properties against P. aeruginosa (PAO1). At 5 mg/mL, the methanolic extract of A. deliciosa exhibited antibacterial activity with an 8 mm zone of inhibition and suppressed PAO1 growth. At 2.5 mg/mL and 1.25 mg/mL doses, PAO1 significantly decreased the production of biofilms by 60% and 29%, respectively. In addition, pyocyanin pigment synthesis was decreased by 30% and 9.25%, respectively, at sub-MIC doses of 2.5 mg/mL and 1.25 mg/mL. When evaluated at 2.5 mg/mL, the extract did not, however, appreciably affect bacterial growth. Conclusion This study enhances the understanding of antibiotic resistance, alternative treatments, and pathogenic microbes. The in vitro findings suggest that A. deliciosa fruit extract may inhibit pyocyanin production in PAO1. Further research with different formulations is recommended to explore its anti-biofilm properties and potential pharmacological applications.

PSLBII-3 Influence of iron supplementation on insulin and glucose dynamics in horses

Abstract Iron is an essential micromineral involved in various physiological functions such as oxygen transport. The National Research Council’s Nutrient Requirements of Horses has iron requirements set at 400 mg iron daily for a 500 kg horse at maintenance; however, a survey found that Thoroughbreds consumed well over the daily requirement at 3,900 mg of iron from hay and grain alone. Additionally, a previous study has found a correlation between hyperinsulinemia and increased ferritin, an indicator of iron body stores. Therefore, the objective of the present study was to determine the effects of iron supplementation on body iron stores as well as insulin and glucose dynamics. It was hypothesized that iron supplementation would influence body iron stores and insulin and glucose responses. Mixed-breed geldings [n = 12; 558.9 ± 74.4 kg body weight (BW)] were housed individually for the study. Horses were fed 2% of their BW in grass hay and an iron-free vitamin mineral supplement, consuming an average of 600 mg of iron daily for the first 28 d (Hay Phase). Horses were then assigned to continue on the hay diet (CTRL; n = 4) or an iron-supplemented diet (IRON; n = 8), in which an oral iron supplement was given daily in the form of ferrous sulfate, with IRON horses consuming an average of 4,000 mg of iron daily for 28 d (Supplement Phase). Oral sugar tests (OSTs) were performed at the beginning of the study as well as the end of each phase, using a 0.45 mL/kg of BW Karo Light Syrup dose to determine insulin response, with jugular venous samples taken prior to the dose and 60 min after. All statistical analysis was performed in GraphPadPrism Version 10.2.0 (GraphPad Software, Boston, MA), using analysis of variance for repeated measures and correlation analysis. All OSTs throughout the study had a significant time effect (P &amp;lt; 0.05) for both insulin and glucose. There was a time X treatment trend (P = 0.1) for insulin during the Supplement Phase and when using multiple comparisons, insulin response was significantly greater (P &amp;lt; 0.0007) at 0 versus 60 min in IRON horses. Although there was no correlation found between 60-min insulin and ferritin concentrations in both IRON and CTRL horses, mean ± SD insulin and ferritin in the IRON group were 23.6 ± 10 uU/mL and 570 ± 195.8 ng/mL, respectively, while in the CTRL group these were 12.7 ± 2.9 uU/mL and 453 ± 31.1 ng/mL, respectively. In conclusion, iron supplementation with an inorganic form at 10 times the daily requirement appears to result in increased insulin responses as well as body iron stores. Further research needs to determine if feeds naturally enriched in iron impact glucose metabolism.

Effect of smoking on the ceramide profile of adipose tissue in patients with cardiovascular diseases

Aim. To describe ceramide profile of subcutaneous, epicardial, perivascular adipose tissue (SAT, EAT, PVAT, respectively) in patients with cardiovascular diseases (CVD) depending on smoking.Material and methods. Ceramide (CER) profile in the SAT, EAT, PVAT of patients with CVD (30 patients with coronary artery disease and 30 patients with degenerative non-rheumatic acquired heart defects) was determined using a high-resolution chromatography mass spectrometry (Agilent 1200 series (Agilent Technologies, Germany)) with a maXis impact mass spectrometer (Bruker, Germany)). Biopsies of SAT, EAT, PVAT were obtained during surgery. Statistical analysis was performed in GraphPad Prism 8 (GraphPad Software).Results. Associations of CER levels in visceral adipose tissue and smoking were found, manifested by an increase in the concentration of CER d18:1/16:0, 18:0, 24:1, 22:0. In the SAT of smoking patients, an increased content of Cer d18:1/14:0, 17:0 was found.Conclusion. The ceramide profile of local fat depots differed in smoking and non-smoking patients with CVD. The obtained results indicate the modulation of CER synthesis during smoking and the accumulation of CER mainly in the EAT and PVAT in CVD. Taking into account the wide range of metabolic effects, the use of CER as biomarkers and targets for personalized therapy is promising for identifying high-risk individuals and improving therapeutic and diagnostic strategies for CVD.

Impact of Previous Surgery and/or Radiation Therapy on Endoscopic Reconstruction Outcomes.

The impact of prior local therapies, including radiation and surgery, on reconstruction outcomes after endonasal surgery is currently not well known. Reconstruction nuances in the preoperative setting merit further evaluation to avoid potential postoperative complications that can hinder overall tumor management and negatively impact patient outcome. We sought to determine whether prior treatments increase risk of reconstruction-related postoperative morbidity and to evaluate the effectiveness of our current treatment paradigm for skull base reconstruction. A retrospective review of all endonasal surgeries for tumor resection between March 2000 and March 2022 was performed. Patients were grouped based on treatment history. Patient demographics, operative, and postoperative reconstruction-related morbidity data were collected, including cerebrospinal fluid leak, sinonasal morbidity, and infectious complications. Variables significantly associated with postoperative complications in the univariate analysis were included in the multivariate Cox proportional hazards regression model. Complication-free survival curves were generated, and the log-rank test evaluated the relationship between complication-free survival and the different clinical, surgical, and treatment parameters. All statistical analyses were performed with SPSS 26 (IBM Corp) and Graph Pad 9.0 (GraphPad Software). A total of 418 patients were included. 291 patients had no prior treatments, 49 patients had previously received radiation, and 78 patients had prior surgeries. Of the 49 patients who had prior radiation, 27% underwent reconstruction with tunneled pericranial flaps vs 16% of treatment-naïve patients. On multivariate analysis, prior treatment was not significantly associated with reconstruction-related complications. Negative smoking history, no leak or small intraoperative leak, and use of vascularized flap in reconstruction were protective factors. In patients undergoing endonasal surgery, prior radiation and/or surgery does not appear to significantly increase the risk of immediate or delayed reconstruction complications using our current reconstructive management plan, which incorporates an upfront regional flap for high-risk cases.

Restoration of Radiosensitivity by Soya Isoflavone Genistein is Accomplished by Facilitating DNA Damage Response in Radioresistant Cervical Cancer in vitro

ABSTRACT Context: Enhancing radiotherapeutic efficacy in tumor cells and sparing the normal tissues are major clinical concerns for the betterment of cancer therapy. Genistein (GEN) being a radiosensitizer ameliorates the effectiveness of radiation-induced cell killing by inducing DNA damage. This molecule is accountable for minimizing radiation-related toxicity and protecting healthy cells. However, the explicit mechanism of action of such molecules needs exploration. Aims: The objective of this study is to investigate the mechanistic action of GEN in cervical cancer cell radiosensitization. Settings and Design: Cervical squamous carcinoma cell SiHa and a radioresistant subline SiHa/RR (developed and isolated from SiHa) were taken for this study. The experiments were performed by pretreating the cells with IC30 dose of GEN, followed by acute irradiation to detect the impact of GEN in imparting radiosensitivity. Subjects and Methods: Optimal dose selection of GEN was performed by MTT assay, and radiosensitizing potency was determined by pretreating the cells with IC30 dose of GEN, followed by challenging with acute incremental doses of radiation. Mechanistic parameters were checked by clonogenic assay, cell cycle analysis, DNA damage estimation, apoptosis, and wound healing-sphere-forming assay. Statistical Analysis Used: Statistical analysis was performed in GraphPad software by performing the Student’s t-test. Results: Results depicted decreased numbers of colonies, increased frequency of DNA damage and apoptotic cells, and suppressed wound healing ability along with restrained sphere-forming ability upon the intervention of cells with GEN before radiation exposure. Such observations implied that GEN pretreatment renders improved radiosensitivity in cervical cancer by increased DNA damage-mediated G2/M arrest with subsequent apoptosis. Conclusions: GEN by inducing DNA damage stimulates radiation-induced cell killing in vitro.

BURN INJURY RESULTS IN MYELOID PRIMING DURING EMERGENCY HEMATOPOIESIS.

Introduction: Hematopoiesis proceeds in a tiered pattern of differentiation, beginning with hematopoietic stem cells (HSC) and culminating in erythroid, myeloid, and lymphoid lineages. Pathologically altered lineage commitment can result in inadequate leukocyte production or dysfunctional cell lines. Drivers of emergency hematopoiesis after burn injury are inadequately defined. Burn injury induces a myeloid predominance associated with infection that worsens outcomes. This study aims to further profile bone marrow HSCs following burn injury in a murine model. Methods: C57BL/6 mice received burn or sham injury with ~12% total body surface area scald burn on the dorsal surface with subsequent sacrifice at 1, 2, 3, 7, and 10 days postinjury. Bone marrow from hindlimbs was analyzed for HSC populations via flow cytometry and analyzed using FlowJo Software (version 10.6). Event counts and frequencies were analyzed with multiple unpaired t tests and linear mixed-effect regression. Real-time polymerase chain reaction performed on isolated lineage-negative bone marrow cell RNA targeted PU.1, GATA-1, and GATA-3 with subsequent analysis conducted with QuantStudio 3 software. Statistical analysis and representation were performed on GraphPad software (Prism). Results: Flow cytometry revealed significantly elevated proportions of long-term HSCs at 3 days post-injury ( P < 0.05) and short-term HSCs at days 2, 3, and 10 (all P < 0.05) in burn-injured mice. There was a sustained, but not significant, increase in proportions in the multipotent progenitor (MPP) 2 and 3 subpopulations in the burn cohort compared to sham controls. The common myeloid progenitor (CMP) proportion was significantly higher on days 3 and 10 (both P < 0.01), whereas the granulocyte-macrophage progenitor (GMP) proportion increased on days 1, 2, and 10 ( P < 0.05, P < 0.01, P < 0.01, respectively). Although the megakaryocyte-erythrocyte progenitor (MEP) proportion appeared consistently lower in the burn cohort, this did not reach significance. mRNA analysis resulted in a downregulation of PU.1 on day 1 ( P = 0.0002) with an upregulation by day 7 ( P < 0.01). GATA-1 downregulation occurred by day 7 ( P < 0.05), and GATA3 showed downregulation on days 3 and 7 ( P < 0.05). Discussion: Full-thickness burn results in an emergency hematopoiesis via proportional increase of long-term HSC and short-term HSC/MPP1 subpopulations beginning in the early postinjury period. Subsequent lineage commitment displays a myeloid predominance with a shift toward myeloid progenitors with mRNA analysis corroborating this finding with associated upregulation of PU.1 and downregulation of GATA-1 and GATA-3. Further studies are needed to understand how burn-induced emergency hematopoiesis may predispose to infection by pathologic lineage selection.

Correlation study: Bone density and circulating inflammatory markers in postmenopausal patients.

This study aims to investigate the correlation between changes in bone mineral density (BMD) in postmenopausal women and circulating inflammatory markers. This retrospective study focused on postmenopausal women admitted to the orthopedic department of Suzhou Benq Medical Center from June 2022 to December 2023, following predetermined inclusion and exclusion criteria. We retrospectively collected data on initial blood routine test results and bone density measurements for all study subjects upon admission, including parameters such as white blood cell count (WBC), C-reactive protein, interleukin-6 (IL-6), and procalcitonin (PCT). Additionally, the systemic immune-inflammation index (SII) was calculated using neutrophil count, lymphocyte count, and platelet count. Statistical analyses using SPSS and GraphPad software were performed to assess the correlation between bone density and inflammatory markers. Patients were classified into three groups based on BMD results, including 60 individuals in the osteoporosis (OP) group, 127 individuals in the osteopenia group, and 37 individuals in the Normalgroup, respectively. Principal component analysis analysis suggested that WBC, SII, and postmenopausal OP(PMOP) held significant feature values. Correlation analysis indicated a correlation between WBC (p = 0.021), IL-6 (p = 0.044), SII (p = 0.034), and PMOP. One-way ANOVA analysis revealed significant differences in IL-6 (p = 0.0179), SII (p = 0.0210), and PCT (p = 0.0200) among the three groups. Finally, ROC curve analysis demonstrated that SII (area under the curve = 0.716) has predictive value for PMOP. This study identified a certain predictive value for PMOP through the assessment of inflammatory markers in peripheral blood using routine blood tests.

Prescribing Trend of Antibiotics Among the Patients Admitted in Intensive Medical Care Unit: A Prospective Observational Study.

Intensive medical care units (IMCUs) usually admit patients who are in critical medical need and require the utmost attention of healthcare professionals, along with the best treatment options available. These patients are prone to infections and require various antibiotics for the treatment. Varying costs of antibiotics, variable dosage forms, and antibiotic resistance cause an economic burden on patients Methodology: This study was designed and conducted prospectively to evaluate the prescribing pattern of antibiotics at the IMCU in a tertiary care hospital. A total of 102 patients were included in the study based on the exclusion and inclusion criteria, and the collected data was tabulated in an Excel sheetand analyzed using Prism GraphPad software. Data were presented as numbers and percentages. Most of the patients were in the age group of 41-50 years. The number of male patients was slightly more than that of female patients. The majority of the patients admitted to the IMCU had acute pulmonary edema and cerebrovascular accidents. Most antibiotics were prescribed empirically and administered parenterally, of which Amoxicillin + Potassium clavulanate was the most commonly used antibiotic. Tigecycline had the highest daily defined dose per 100 bed days value, and injection Sulbactam + Cefoperazone was the costliest of all antibiotic therapy. Antibiotic therapy used in the IMCU consisting of Sulbactam + Cefoperazone was found to be costlier, and Amoxicillin + Potassium clavulanate were the commonly prescribed antibiotics among the other prescribed antibiotics. The average cost of antibiotics was found to be higher, which increased the economic healthcare burden for patients and their families.

The Increased Frequency of Type 1 Regulatory T (Tr1) Cells and the Altered Expression of Aryl Hydrocarbon Receptor (AHR) and Interferon Regulatory Factor-4 (IRF4) Genes in Type 1 Diabetes: A Case-Control Study.

Background and aim Type 1 diabetes is an autoimmune disorder characterized by the destruction of pancreatic beta cells, leading to insulin deficiency and hyperglycemia. Regulatory T cells (Tregs), particularly type 1 regulatory T (Tr1) cells, play a crucial role in modulating autoimmune responses. Therefore, this study aimed to evaluate the frequency of Tr1 cells and their association with aryl hydrocarbon receptor (AHR) and interferon regulatory factor-4 (IRF4) gene expression levels in type 1 diabetes mellitus (T1DM) compared to the healthy controls. Method A case-control study design was used. The case group included patients diagnosed with T1DM, while the control group consisted of healthy individuals, matched for age and sex. Blood samples were collected, and peripheral blood mononuclear cells (PBMCs) were isolated. Serum interleukin 10 (IL-10) and interleukin 21 (IL-21) levels were measured using enzyme-linked immunosorbent assay (ELISA). The gene expression of AHR and IRF4 was analyzed using quantitative real-time polymerase chain reaction (qPCR), and Tr1 cell populations were determined using flow cytometry. Data were summarized with mean and standard error of the mean (SEM) for quantitative variables. Independent sample t-test, chi-square test, and the Mann-Whitney U test were used to compare groups. Statistical analyses were performed using SPSS version 25 (IBM SPSS Statistics, Armonk, NY), with significance levels set at p < 0.05. Figures were created using GraphPad Prism (GraphPad Software, San Diego, CA). Results A total of 45 cases were enrolled in the study, with 30 T1DM patients and 15 healthy controls. The mean IL-10 concentration was significantly higher in the patients (10.4 ± 1.1 pg/mL) compared to the healthy controls (5.1 ± 0.7 pg/mL), with a p-value of 0.001. There was no significant difference in IL-21 levels between the patients (76.1 ± 9.0 pg/mL) and healthy controls (88.2 ± 17.5 pg/mL), indicated by a p-value of 0.480. AHR gene expression was significantly lower in patients, with a p-value of 0.037. Although IRF4 gene expression was higher in patients, the difference was not statistically significant (p= 0.449). Tr1 cell frequency was significantly higher in T1DM patients (1.45% of cluster of differentiation 4+ {CD4+} T cells) compared to the healthy controls (0.40% of CD4+ T cells), with a p-value of 0.045. Conclusions The study demonstrated that T1DM is associated with higher IL-10 levels, decreased AHR gene expression, and a higher frequency of Tr1 cells. Policymakers should focus on developing targeted immunomodulatory therapies to address these immunological abnormalities. Healthcare providers should prioritize monitoring cytokine levels and gene expression in T1DM patients to tailor treatment plans effectively. Further research is needed to explore the therapeutic potential of modulating Tr1 cells and their related pathways in T1DM management.