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Background: Data available support that ninety percent of male infertility cases are due to low sperm counts. There is a scarcity of data on the medicinal effects of cannabis on fertility. This study evaluated testicular function and sperm quality modulation with cannabis in rats., Methodology: Twenty-five male Wistar rats were randomly grouped into five: A, B, C, and D, each group have 5 rats. A (control): 0.2 ml 2% DMSO, B (vitamin C): 90 mg/kg body weight, C, D, and E were administered: 5 mg/kg, 10 mg/kg and 20 mg/kg body weight of ethanolic leaf extract of cannabis (ELEC) respectively. The rats were sacrificed 24 h after the last day of the 60 day oral administrations. Flavonoids were the predominant phytochemical present in the extract while quercetin, kemferol, silyman and gallic acid were identified., Results: The results showed a significant improvement (p < 0.05) in sperm quality and a significant increase in the concentrations of follicle-stimulating hormone, luteinizing hormone, triglycerides, cholesterol, and total protein determination compared to the normal control. Similarly, there was a significant increase (p < 0.05) in the activities of acid phosphatase, alkaline phosphatase, and superoxide dismutase compared to the normal control. RAC-alpha serine/threonine-protein kinase (AKT1)-silymarin complexes (-8.30 kcal/mol) and androgen receptor (AR)-quercetin complexes (9.20 kcal/mol) had the highest affinity., Conclusion: The antioxidant effects of the flavonoids in the ethanolic extract of cannabis may have protected testicular and sperm cells from oxidative damage. Biochemical processes and histopathological morphology were preserved by cannabis. The docking prediction suggests that the bioactive principle of cannabis may activate the androgenic receptors. The androgenic receptor modulation may be attributed to silymarin and quercetin., (© 2022. The Author(s).)

Background: Hepatitis B infection is a public health concern globally. HBV can be associated with type II diabetes mellitus, as HBV outbreaks have been observed among diabetics in healthcare facilities. This study evaluates the prevalence of HBV infection among patients with type II diabetes mellitus., Method: A total of one hundred and eighty (180) diabetic patients and one-hundred non-diabetics (Controls) were recruited for this study. Structured questionnaires were administered to the consented participants to obtain relevant data. Sera samples obtained were screened using the HBsAg ELISA kit; CTK Biotech, Inc, while the 5 panel kit-rapid diagnostic test, was used to assay for serological markers. Questionnaires were used to obtain relevant information and demographic data., Result: Overall prevalence of HBV infection among diabetes patients was 13.3%. Breakdown showed 9 (5.0%) seropositivity was obtained among male subjects compared to 15(8.3%) recorded among the females, P = .834; P < .05. Subjects aged 41-50 years recorded, 7(3.9%) positivity P = .774; P > .05. Educational status of participants showed 22 (12.2%) positivity among subjects with tertiary level of education P = .032; P < .05). Risk factors considered showed that 5(2.8%).seropositive subjects were alcoholic consumers (P value = .9711; P > .05). Result among non-diabetics (Control) subjects showed (4%) seropositivity among the male subjects compared to (5.0%) seropositivity recorded among the female subjects (P = .739; P > .05)., Conclusion: There is an indication of higher risk of HBV infection among type 2 diabetic patients when compared to non-diabetics. There is the need for more research on this area of study, to further validate the association between HBV infection and Diabetes Mellitus., (© 2021 The Authors. Journal of Clinical Laboratory Analysis Published by Wiley Periodicals LLC.)

Epidemiologic studies have established a relationship between pediatric patients and typhoid fever infection. This study was carried out to ascertain if specific hematological measurements of the pediatric patients discriminate between their positive and negative status to typhoid infection and to produce a rule for classifying other pediatric patients. Discriminant analysis was applied to predict the probability of a specific categorical outcome based on several explanatory variables (predictors). This study analyzed the differentiation between two hundred pediatric patients attending Landmark University Medical Centre based on their typhoid fever status. The hematological parameters considered were Packed Cell Volume, White Blood Cell count; Neutrophil, Erythrocyte level, Hemoglobin and Platelet count, Assay of samples were performed using standard procedures. Fisher's Linear Discriminant Method was used for classification of variables in this study. With the use of the Fisher's Linear Discrimination method for classification of the obtained data, a minimum value of -0.0067 was obtained implying that any new pediatric patient with a discriminant score above -0.0067 would be diagnosed to be typhoid negative; otherwise, they would be classified as typhoid positive pediatric patients. The efficiency of this method of classification was tested using two approaches; Retribution estimate approach and leaving-one out approach which showed a prevalence rate of typhoid positive patients at 75.8% and 74.7% respectively. This data analysis hypotheses that typhoid fever is highly endemic amongst our study subjects. A point-of-care diagnosis with a strong positive predictive value, which improves pediatric enteric fever diagnosis, is strongly advocated., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Enteric fever is an invasive bacterial infection mostly caused by Salmonella enterica serovar Typhi, which is a common agent of enteric fever. This illness has been a major public health issue, as it affects a large number of individuals globally. The box-plot analytic method is involved in exploratory data analysis using statistical techniques to identify patterns that may be hidden in a group of numbers used to visually summarize and compare groups of data. We evaluted the effect of enteric fever on various haematologic parameters using the box-plot distribution model. Samples were obtained from 400 volunteer patients as well as healthy subjects (controls). Assay for typhoid fever was carried out using obtained serum samples to detect specific O and H antigens. Antibody titres of 1:80 and higher for anti-TO and 1:160 and higher for anti-TH antibodies were taken as cutoff values to indicate recent infection of typhoid fever. The haematologic parameters were evaluated using an automated haematology analyser. A statistically significant decrease was observed in packed cell volume, white blood cell count, erythrocyte sedimentation rate and haemoglobin concentration, while a statistically insignificant difference was observed in the neutrophils, lymphocytes and monocytes seen in the box-plot distribution analysis. Typhoid fever causes significant haematologic changes which could be helpful in diagnosis. The box-and-whisker plots compared the distributions of the haematologic parameters, spread and overall ranges. Awareness of these parameters could be useful in providing accurate diagnosis and therapy, particularly in underresourced endemic regions in developing countries., (© 2020 The Authors.)

For longer than a century, the "meningitis belt" of sub-Saharan Africa has experienced the largest-ever global meningitis epidemic. Whereas HIV-associated immunosuppression drives higher susceptibility to environmental infectious organisms with tropism for the central nervous system (CNS), most diagnostic laboratories in the belt stick to  N. meningitidis , H. influenzae , and S. pneumoniae . Cryptococcus neoformans has been the leading cause of death (incidence, 89%; death, 75%). To establish whether diagnostic services target geographically important pathogens, there is a need to know the current spectrum of etiology. Given Africa's agro-silvo-pastoralism, the One Health diagnostic approach is recommended. Considering  multipathogen detection capacity, needed speed for corticosteroid therapy decision, and susceptibility/resistance to antimicrobials with improved CNS penetration, proposed laboratory categorization will help neurologists to choose suitable services.

The genus Staphylococcus includes pathogenic and non-pathogenic facultative anaerobes. Due to the plethora of virulence factors encoded in its genome, the species Staphylococcus aureus is known to be the most pathogenic. S. aureus strains harboring genes encoding virulence and antibiotic resistance are of public health importance. In clinical samples, however, pathogenic S. aureus is often mixed with putatively less pathogenic coagulase-negative staphylococci (CoNS), both of which can harbor mecA, the genetic driver for staphylococcal methicillin-resistance. In this chapter, the detailed practical procedure for operating a real-time pentaplex PCR assay in blood cultures is described. The pentaplex real-time PCR assay simultaneously detects markers for the presence of bacteria (16S rRNA), coagulase-negative staphylococcus (cns), S. aureus (spa), Panton-Valentine leukocidin (pvl), and methicillin resistance (mecA).

Background: Staphylococcal toxicity and antibiotic resistance (STAAR) have been menacing public health. Although vancomycin-resistant Staphylococcus aureus (VRSA) is currently not as widespread as methicillin-resistant S. aureus (MRSA), genome evolution of MRSA into VRSA, including strains engineered within the same patient under anti-staphylococcal therapy, may build up to future public health concern. To further complicate diagnosis, infection control and anti-microbial chemotherapy, non-sterile sites such as the nares and the skin could contain both S. aureus and coagulase-negative staphylococci (CoNS), either of which could harbour mecA the gene driving staphylococcal methicillin-resistance and required for MRSA-VRSA evolution., Results: A new heptaplex PCR assay has been developed which simultaneously detects seven markers for: i) eubacteria (16S rRNA), ii) Staphylococcus genus (tuf), iii) Staphylococcus aureus (spa), iv) CoNS (cns), v) Panton-Valentine leukocidin (pvl), vi) methicillin resistance (mecA), and vii) vancomycin resistance (vanA). Following successful validation using 255 reference bacterial strains, applicability to analyse clinical samples was evaluated by direct amplification in spiked blood cultures (n = 89) which returned 100 % specificity, negative and positive predictive values. The new assay has LoD of 1.0x10(3) CFU/mL for the 16S rRNA marker and 1.0x10(4) CFU/mL for six other markers and completes cycling in less than one hour., Conclusion: The speed, sensitivity (100 %), NPV (100 %) and PPV (100 %) suggest the new heptaplex PCR assay could be easily integrated into a routine diagnostic microbiology workflow. Detection of the cns marker allows for unique identification of CoNS in mono-microbial and in poly-microbial samples containing mixtures of CoNS and S. aureus without recourse to the conventional elimination approach which is ambiguous. In addition to the SA-CoNS differential diagnostic essence of the new assay, inclusion of vanA primers will allow microbiology laboratories to stay ahead of the emerging MRSA-VRSA evolution. To the best of our knowledge, the new heptaplex PCR assay is the most multiplexed among similar PCR-based assays for simultaneous detection of STAAR.

Staphylococcus aureus strains harbouring genes encoding virulence and antibiotic resistance are of public health importance. In clinical samples, pathogenic S. aureus is often mixed with putatively less pathogenic coagulase-negative staphylococci (CoNS), both of which can harbour mecA, the gene encoding staphylococcal methicillin-resistance. There have been previous attempts at distinguishing MRSA from MRCoNS, most of which were based on the detection of one of the pathognomonic markers of S. aureus, such as coa, nuc or spa. That approach might suffice for discrete colonies and mono-microbial samples; it is inadequate for identification of clinical specimens containing mixtures of S. aureus and CoNS. In the present study, a real-time pentaplex PCR assay has been developed which simultaneously detects markers for bacteria (16S rRNA), coagulase-negative staphylococcus (cns), S. aureus (spa), Panton-Valentine leukocidin (pvl) and methicillin resistance (mecA). Staphylococcal and non-staphylococcal bacterial strains (n = 283) were used to validate the new assay. The applicability of this test to clinical samples was evaluated using spiked blood cultures (n = 43) containing S. aureus and CoNS in mono-microbial and poly-microbial models, which showed that the 5 markers were all detected as expected. Cycling completes within 1 h, delivering 100% specificity, NPV and PPV with a detection limit of 1.0 × 10(1) to 3.0 × 10(1) colony forming units (CFU)/ml, suggesting direct applicability in routine diagnostic microbiology. This is the most multiplexed real-time PCR-based PVL-MRSA assay and the first detection of a unique marker for CoNS without recourse to the conventional elimination approach. There was no evidence that this new assay produced invalid/indeterminate test results., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Staphylococcus aureus produces several toxins, including Panton-Valentine leukocidin (PVL). The involvement of PVL in primary skin infections, necrotizing pneumonia, musculoskeletal disorders, brain abscess, and other diseases, some of which are life-threatening, has been reported. Following expert opinion, we aimed to provide the tools for establishment of sequence-based diagnostics and therapeutics for those conditions. We engineered the synergistic S and F (LukS-PV and LukF-PV respectively) pro-toxin subunits from Staphylococcus aureus USA400 into separate expression E. coli BL21(DE3)-pLysS hosts., Results: Following Nickel affinity chromatography (NAC), the F subunit came out without bands of impurity. The S sub-unit did not come off very pure after NAC thus necessitating further purification by size exclusion and ion-exchange chromatography. The purification plots showed that the BioLogic-LP and AKTA systems are reliable for following the progress of the chromatographic purification in real-time. Computer predicted Mw for the 6His-LukF-PV and 6His-LukS-PV were 35645.41 Da and 33530.04 Da respectively, while the mass spectrometry results were 35643.57 Da and 33528.34 Da respectively., Conclusion: The BioLogic-LP and AKTA systems are commendable for reliability and user-friendliness. As a recent work elsewhere also reported that a second round of chromatography was necessary to purify the S subunit after the first attempt, we speculate that the S subunit might contain yet unidentified motif(s) requiring further treatment. The purified S and F sub-units of PVL were supplied to the Nottingham Cancer Immunotherapy group who used them to establish sequence-based monoclonal antibodies for diagnostic and therapeutic uses targeting PVL.