Exploring the multifaceted effects of Ammi visnaga: subchronic toxicity, antioxidant capacity, immunomodulatory, and anti-inflammatory activities.

Ammi visnaga (A. visnaga) is an annual herb that has been used in traditional medicine to treat various ailments attributed to the presence of its bioactive compounds. The purpose of this study was to identify and examine the phytochemical properties of the hydroalcoholic extract of A. visnaga using in vitro and in vivo models. Our findings demonstrated that the extract contained a variety of beneficial components, including phenols, flavonoids, tannins, coumarins, saponins, khellin, and visnagin. The total polyphenolic content and total flavonoid content were 23.26 mg/GAE/g dry weight and 13.26 mg/GAE/g dry weight, respectively. In vitro tests demonstrated that the extract possessed antioxidant properties as evidenced by the ability to scavenge free radicals, including DPPH, ABTS, nitric oxide (NO), phosphomolybdate, and ferric-reducing antioxidant power (FRAP). Further, the extract was found to inhibit hydrogen peroxide (H2O2)-induced hemolysis. In a 90-d in vivo study, female Wistar rats were administered 1 g/kg of A. visnaga extract orally resulting in a significant increase in total white blood cell count. Although morphological changes were observed in the liver, no marked alterations were noted in kidneys and spleen. In a female Swiss albino mice model of acetic acid-induced vascular permeability, A. visnaga significantly inhibited extravasations of Evans blue at doses of 0.5 or 1 g/kg with inhibition percentages of 51 and 65%, respectively, blocking tissue necrosis. The extract also demonstrated potential immunomodulatory properties in mice by enhancing antibody production in response to antigens. In silico molecular docking studies demonstrated a strong affinity between khellin or visnagin and immunomodulatory proteins, NF-κB, p52, and TNF-α. These findings suggest that A. visnaga may be considered a beneficial antioxidant with immunomodulatory properties and might serve as a therapeutic agent to combat certain diseases.

Single-cell RNA-Seq reveals intracellular microbial diversity within immune cells during SARS-CoV-2 infection and recovery.

Intracellular microorganisms, like viruses, bacteria, and fungi, pose challenges in detection due to their non-culturable forms. Transcriptomic analysis at cellular level enables exploration of distributions and the impact of these microorganisms on host cells, a domain that remains underexplored because of methodological limitations. Single-cell technology shows promise in addressing this by capturing polyadenine-tailed transcripts, because recent studies confirmed polyadenylation in microbial transcriptomes. We utilized single-cell RNA-seq from PBMCs to probe intracellular microbes in healthy, SARS-CoV-2-positive, and recovered individuals. Among 76 bacterial species detected, 16 showed significant abundance differences. Buchnera aphidicola, Streptomyces clavuligerus, and Ehrlichia canis emerged significantly in memory-B, Naïve-T, and Treg cells. Staphylococcus aureus, Mycoplasma mycoides, Leptospira interrogans, and others displayed elevated levels in SARS-CoV-2-positive patients, suggesting possible disease association. This highlights the strength of single-cell technology in revealing potential microorganism's cell-specific functions. Further research is essential for functional understanding of their cell-specific abundance across physiological states.

Evaluation of oxidative stress markers in Rwanda during the SARS-CoV-2 pandemic: A cross-sectional study.

COVID-19 is mainly described as endothelial dysfunction, and due to the bidirectional link between oxidative stress and endothelial dysfunction, we initiated a program directed to the evaluation of the oxidative status of the population of Rwanda by measuring spectrophotometrically their plasma Reactive Oxygen Metabolites (d-ROMs) and Plasma Antioxidant Potential (PAT). The reference population was chosen to reflect the absence of actual or past SARS-CoV-2 infections as well as other clinically established infective status and reference intervals for d-ROM and PAT were identified. The average d-ROM was 378.6 UCARR with a standard deviation of 105.2, a value significantly higher than that reported for Caucasian or East Asian population (260-300 UCARR). The average PAT value was 2853.6, with a standard deviation of 635.7 UCOR, at the upper limit according to the averaged values for healthy Caucasian populations. The results of this study, the first so far reported on a sub-Saharan population, can effectively be used as a baseline value for clinical management of inflammatory conditions, for the stratification of at-risk individuals and to inform recommendations for effective use of public health resources.

Correlation of Malaria Rapid Test and Peripheral Blood Smear Microscopy among Patients attending Byumba Health Centre.

Background: Malaria presents a diagnostic challenge in most tropical countries including Rwanda. Microscopy remains the gold standard for diagnosing malaria, however, it is labour intensive and depends upon the skill of the examiner. Malaria rapid diagnostic tests (MRDTs) have been developed as an easy, convenient alternative to microscopy. Methods: A cross sectional study was conducted from October to November 2019 on 130 febrile patients who were directed to the laboratory department for blood screening for malaria parasites at Byumba Health centre. The main objective of this study was to correlate Microscopy and MRDTs in diagnosis of malaria. Results: After signing a consent form, blood samples were collected and screened for malaria parasites microscopically and by using MRDTs. Data collection forms were filled with relevant information and obtained results for MRDTs and for peripheral blood smear were recorded. The collected data were statistically analyzed using GraphPad Prism 9 software. The mean age found to be 16 years old. In this study peripheral blood smear microscopy was considered as a reference method. The sensitivity and specificity of RDT Histidine-Rich Protein 2 (HRP-2) were calculated and found to be 96.6% and 60% respectively. The negative predictive value was found to be 92.85% where positive predictive value was 73.3%. Conclusion: MRDTs should be used along with microscopy to avert complications associated with delayed diagnosis and similar studies are required to identify alternative techniques with high specificity for the diagnosis of malaria.

Bacteriological Profile and Antimicrobial Susceptibility Patterns of Bloodstream Infection at Kigali University Teaching Hospital.

BACKGROUND: Worldwide, bacterial bloodstream infections (BSIs) constitute an important cause of morbidity and mortality in clinical settings. Due to the limited laboratory facilities in sub-Saharan Africa, poor diagnosis of BSIs results in poor clinical outcomes and leads to a risk of antimicrobial resistance. The present work was carried out to describe the microbiological features of BSIs using the data collected from Centre Hospitalier Universitaire de Kigali (CHUK). METHODS: A retrospective study was carried out at CHUK. The blood culture results of 2,910 cases - from adults, children and infants - were reviewed in the Microbiology service from October 2017 to October 2018. The following variables were considered: age, gender, admitting department, blood culture results, and antimicrobials sensitivity test results. Data were entered and analyzed using Microsoft Excel 2013. RESULTS: Twelve percent (341/2,910) of blood culture results reviewed were positive with 108 (31.7%) Gram positive bacteria and 233 (68.3%) Gram negative bacteria. The most prevalent pathogens were Klebsiella pneumoniae 108 (31.7%) and Staphylococcus aureus 100 (29.3%). This study revealed a high resistance to commonly prescribed antibiotics such as penicillin, trimethoprim sulfamethoxazole, and Ampicillin with 91.8, 83.3, and 81.8% of resistance, respectively. However, bacteria were sensitive to imipenem and vancomycin with 98.1 and 94.3% of sensitivity, respectively. The pediatrics and neonatology departments showed a high number of positive culture with 97/341 (28.4%), and 93/341 (27%) respectively. The overall prevalence of multidrug resistance was 77.1%. CONCLUSION: The prevalence of bacterial pathogens in BSIs was found to be high. The antibiotic resistance to the commonly used antibiotics was high. Appropriate treatment of BSIs should be based on the current knowledge of bacterial resistance pattern. This study will help in formulating management of diagnostic guidelines and antibiotic policy.

Bacteriological Study of Electronic Devices Used by Healthcare Workers at Ruhengeri Referral Hospital.

Electronic devices have become one of the most essential accessories being used in hospitals. Those devices increase the communication and contact making healthcare delivery more efficient and quality service oriented. The study was designed to collect reliable information about the spreading of pathogens through electronic devices especially in sensitive departments. The objectives of this study were to evaluate the bacterial colonization of electronic devices and determine the effectiveness of disinfection with alcohol 70% (w/v) to reduce the bacterial colonization of electronic devices. It was a cross-sectional study where samples were collected by means of moistened swabs in sterile saline solution from 30 electronic devices used by healthcare workers at Ruhengeri Referral Hospital within four different units: maternity, neonathology, intensive care, and theater room. To evaluate the effects of disinfection using 70% isopropyl alcohol, the second sample collection was carried out after decontamination with 70% isopropyl alcohol. Samples were analyzed in the microbiology lab of INES-Ruhengeri. The result showed that Staphylococcus aureus was the most predominant with 22.5%. Lactobacillus and Citrobacter spp. were 12.5%; Pseudomonas aeruginosa, coagulase-negative Staphylococci, and Serratia marcescens were 10%; Escherichia coli was 7.5%; Klebsiella spp. and Providencia spp. were at 5%. The lowest prevalence was 2.5% of Enterobacter spp. and Salmonella spp. The threat of dissemination of isolated microorganisms is valid, since all devices evaluated in this study showed bacterial contamination of species associated to hospital-acquired infections. Special care should be taken when using electronic devices in healthcare settings in addition to disinfection to reduce the risk of transmission of bacterial agents. Further studies should evaluate the antibiotic susceptibility for better conclusive results since all isolated bacteria in this study were subjected to high resistance and were associated with nosocomial infections.

Risk factors associated with hepatitis B and C in rural population of Burera district, Rwanda.

INTRODUCTION: Several studies have shown that older people have a higher risk of exposure to viral hepatitis B and C than younger people. This study aimed to determine the seroprevalence of hepatitis B and C and their associated factors in people aged 45+ years old in Burera, a rural district of Rwanda. METHODS: A cross sectional study was conducted from July to December 2017 during a mass campaign of hepatitis B (HBV) and hepatitis C (HCV) screening and vaccination of eligible populations against HBV in Burera District. Blood samples were collected and hepatitis B surface antigen (HBsAg) and an antibody against hepatitis C (Anti-HCV) were detected using an Enzyme-Linked Immuno-Sorbent Assay (ELISA). The associated factors were identified using a structured questionnaire and the data was analyzed using SPSS software. RESULTS: Of the 374 people included in this study, 53.2% were females. The median age was 56 years old with an interquartile range (IQR) of 50 - 63 years old. The prevalence of HBV and HCV infection was 6.4% and 9.4%, respectively, with 0.3% co-infection rate. Age, social economic level, history of blood transfusion, history of never using a condom, as well as a history of injury with a used sharp material were significantly associated with HCV infection. CONCLUSION: The study showed a high seroprevalence of both HBV and HCV in Burera's elderly population aged 45+ years. Several factors associated with HBV and HCV in this study could be prevented through education and improved hygiene.

Risk factors associated with hepatitis B and C in rural population of Burera district, Rwanda

Introduction: several studies have shown that older people have a higher risk of exposure to viral hepatitis B and C than younger people. This study aimed to determine the seroprevalence of hepatitis B and C and their associated factors in people aged 45+ years old in Burera, a rural district of Rwanda.Methods: a cross sectional study was conducted from July to December 2017 during a mass campaign of hepititis B (HBV) and hepititis C (HCV) screening and vaccination of eligible populations against HBV in Burera District. Blood samples were collected and hepatitis B surface antigen (HBsAg) and an antibody against hepatitis C (Anti-HCV) were detected using an Enzyme-Linked Immuno-Sorbent Assay (ELISA). The associated factors were identified using a structured questionnaire and the data was analyzed using SPSS software.Results: of the 374 people included in this study, 53.2% were females. The median age was 56 years old with an Interquartile range (IQR) of 50 - 63 years old. The prevalence of HBV and HCV infection was 6.4% and 9.4%, respectively, with 0.3% co-infection rate. Age, social economic level, history of blood transfusion, history of never using a condom, as well as a history of injury with a used sharp material were significantly associated with HCV infection.Conclusion: the study showed a high seroprevalence of both HBV and HCV in Burera's elderly population aged 45+ years. Several factors associated with HBV and HCV in this study could be prevented through education and improved hygiene

Association between glutathione peroxidase 1 codon 198 variant and the occurrence of breast cancer in Rwanda.

BACKGROUND: Glutathione peroxidase 1 gene (GPX1) is one of the antioxidant enzyme that remove the reactive oxygen species in a continuous process. Since the identification of a well-characterized functional polymorphism named p.Pro198Leu (rs1050450 C>T) in GPX1 gene, abundant studies have evaluated the association between p.Pro198Leu polymorphism and tumor risk in diverse population. But, the available results related to breast cancer are conflicting and absent in Africa. The present case-control study was planned to assess the presence of GPX1 Pro198Leu polymorphism in Rwanda population to determine whether it is associated with the risk of developing breast cancer. METHODS: Genomic DNA from peripheral blood leukocytes of 41 patients with breast cancer and 42 healthy controls were enrolled and genotyped GPX1 Pro198Leu polymorphism by PCR amplification and DNA sequencing. RESULTS: No significant difference in the frequencies of Pro/Pro (49%) and Pro/Leu (51%) genotypes in cancer cases and in controls (50% each) were found. The allelic frequencies of Pro and Leu were 74% versus 26% and 75% versus 25% in breast cancer cases and controls respectively. No association was observed in allele frequencies of Pro and Leu, and familial history. Only an overall association of GPX1 Pro198Leu with grade of cancer (Pro/Leu vs. Pro/Pro: p = .0200) was detected. CONCLUSION: The result of this study suggested that GPX1 Pro198Leu polymorphism could not be a risk factor for breast cancer in Rwanda. However, large-scale studies on the effect of this polymorphism on the factors disturbing the redox homeostasis are needed for conclusive understanding.

CHEK2 Germ Line Mutations are Lacking among Familial and Sporadic Breast Cancer Patients in Rwanda

Worldwide, breast cancer is the most frequent neoplasm and the second leading cause of cancer death among females. It dominates in both developed and developing countries and represents a major public health problem. The etiology is multifactorial and involves exogenous agents as well as endogenous factors. Although they account for only a small fraction of the breast cancer burden, mutations in the BRCA1 and BRCA2 genes are known to confer a high risk predisposition. Mutations in moderate/low-penetrance genes may also contribute to breast cancer risk. Previous studies have shown that mutations in the CHEK2 gene are involved in breast cancer susceptibility due to its impact on DNA repair processes and replication checkpoints. This study was conducted to evaluate the frequencies of three germline mutations in CHEK2 gene (c.1100delC, R145W and I157T) in breast cancers in Rwanda. Using direct DNA sequencing, we analyzed 41 breast cancer patients and 42 normal breast controls but could not detect any positives. CHEK2 mutations may be a rare event in Rwandan population and may only play a minor if an role in breast cancer predisposition among familial and sporadic cases.

Detection of human papillomavirus DNA in tumors from Rwandese breast cancer patients.

BACKGROUND: During the last decades, a great interest was given to viral etiology of breast cancer. Indeed, due to recent technical improvements and some encouraging new results, it has been a resurgence of interest in the possibility that a substantial proportion of human breast cancers may be caused by viral infections. High-risk genotypes of human papillomavirus (HPV) have been found in breast cancer cases. In the present study, we aimed to assess the presence of HPV DNA in breast cancer cases from Rwanda and to evaluate the association between HPV infection and clinico-pathological features. METHODS: Therefore, a total of 47 archived formalin-fixed paraffin-embedded biopsies were collected and complete information was recorded. HPV detection and genotyping were done by PCR amplification and DNA sequencing. RESULTS: Overall, HPV DNA was found in 46.81% of cases, HPV16 being the most prevalent subtype (77.27%) followed by HPV33 (13.64%) and HPV31 (9.09%). Comparison of HPV with clinico-pathological features showed no significant difference between HPV infection and breast localization, histological subtype, clinical stage, tumor grade, and intrinsic molecular subtypes. CONCLUSIONS: These findings provide evidence of high prevalence of high-risk HPV in Rwandese patients with breast cancer and suggest that high-risk HPV infections could be a risk factor associated with human breast cancer development.

Association of p53 Codon 72 Polymorphism with Breast Cancer in a Rwandese Population.

BACKGROUND AND AIMS: A common polymorphism in the tumor suppressor gene p53 at codon 72 has been suggested to play a role in the development of a number of cancers. This polymorphism has been studied in many populations worldwide, with conflicting results. The present study was planned to assess the association of p53 codon 72 polymorphism with breast cancer development in a Rwandese population. METHODS: In this study, the polymorphism was examined by allele-specific PCR analysis in 40 patients with breast cancer and 39 healthy controls. RESULTS: The heterozygous genotype Pro/Arg prevailed in both breast cancer patients and controls, and was present in 80% (32/40) and 92.3% (36/39) of cases, respectively. No statistically significant association was observed between p53 codon 72 polymorphism and breast cancer risk. Distribution of p53 genotypes was also studied according to familial history, tumor grade, and clinical stage, and results clearly showed no statistically significant difference. CONCLUSION: These results suggest that p53 codon 72 polymorphism could not be assessed as a risk factor marker for predisposition to breast cancer in Rwanda. However, further studies using larger sample sizes are needed to provide more conclusive results and to investigate other genetic mutations affecting the activity of p53.