Department of Biochemistry and Biotechnology

Permanent URI for this collectionhttps://koha2.tukenya.ac.ke/handle/123456789/1396

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    South-to-south mentoring as a vehicle for implementing sustainable health security in Africa
    (2023-03-07) Okanya, Patrick W.; Norlock, Stephanie Marie; Trataris, Anastasia
    Background: While sustainability has become a universal precept in the development of global health security systems, supporting policies often lack mechanisms to drive policies into regular practice. ‘On-paper’ norms and regulations are to a great extent upheld by frontline workers who often lack the opportunity to communicate their first-hand experiences to decisionmakers; their role is an often overlooked, yet crucial, aspect of a sustainable global health security landscape. Initiatives and programs developing transdisciplinary professional skills support the increased bidirectional dialogue between these frontline workers and key policy- and decisionmakers which may sustainably narrow the gap between global health security policy design and implementation. Methods: The International Federation of Biosafety Associations’ (IFBA) Global Mentorship Program recruits biosafety and biosecurity champions across Africa to provide local peer mentorship to developing professionals in their geographic region. Mentors and mentees complete structured one year program cycles, where they are provided with written overviews of monthly discussion topics, and attend optional virtual interactive activities. Feedback from African participants of the 2019–2020 program cycle was collected using a virtual Exit Survey, where aspects of program impact and structure were assessed. Results: Following its initial call for applications, the IFBA Global Mentorship Program received considerable interest from professionals across the African continent, particularly in East and North Africa. The pilot program cycle matched a total of 62 individuals from an array of professional disciplines across several regions, 40 of which were located on the African continent. The resulting mentorship pairs shared knowledge, skills, and experiences towards translating policy objectives to action on the front lines. Mentorship pairs embraced multidisciplinary approaches to harmonize health security strategies across the human and animal health sectors. South-to-South mentorship therefore provided mentees with locally relevant support critical to translation of best technical practices to local capacity and work. Conclusion: The IFBA’s South-to-South Global Mentorship Program has demonstrated its ability to form crucial links between frontline biosafety professionals, laboratory workers, and policy- and decision-makers across several implicated sectors. By supporting regionally relevant peer mentorship programs, the gap between health security policy development and implementation may be narrowed.
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    Coenzyme Q10 Ameliorates potassium cyanide-induced toxicosis in a mouse model
    (2023-03-07) Okanya, Patrick W.; Biwott, Kipchumba; Gitaua, Grace Wairimu
    Potassium cyanide (KCN) is one of the most lethal and feared poison; which devastates cellular respiration resulting in death due to hypoxia. Several antidotes exist, but most face major limitations of safety and efficacy. Moreover, there is a need for new strategies to minimize post-exposure pathological sequel, which includes harmful oxidant and in- flammatory changes. Coenzyme Q10 (CoQ10) is a powerful antioxidant, which has shown efficacy against chemical-induced toxicity. In the present study, the potential protective ef- fect of CoQ10 against KCN-induced toxicosis was evaluated. Female Swiss white mice (3–4 weeks old) were divided into three treatment groups. The first group was used as the con- trol, the second group was supplemented with 200 mg/kg of CoQ10 for one month before administration with 8 mg/kg of KCN. For this group, co-administration of CoQ10 and KCN was continued to the end of the experiment. The third group was administered 8 mg/kg of KCN. The experiment was terminated after 42 days post-treatment to enable investiga- tions into the effect of KCN and CoQ10 on various physiological, biochemical, and cellular processes. The results of this study showed that KCN severely impaired the health of mice, more so, the neurological integrity. KCN-driven depletion of cellular glutathione (GSH) was noted in the liver and brain. This constitutes a characteristic impairment of the antioxidant capacity due to the induction of severe oxidative stress. CoQ10 significantly reinforced the neurological integrity and restored cellular glutathione (reduced form) in both the liver and brain, a clear indication of reduced oxidative stress. Remarkably, KCN-induced ane- mia, leukocytosis, and suppression of platelets were reversed by CoQ10 supplementation. Moreover, histopathological analysis revealed that CoQ10 supplementation blocked KCN- driven liver, kidney, and brain inflammation, and characteristic hypoxia-induced lesions. These findings open possibilities for further scrutiny and development of adjunct therapy utilizing CoQ10 to treat KCN poisoning.
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    Screening and Characterization of Hydrocarbonoclastic Bacteria Isolated from Oil-contaminated Soils from Auto Garages
    (2023-03-06) Okanya, Patrick; Mbatia, Betty Nyambura; Muge, Edward Kirwa
    Release of petroleum oil and its products into the environment is a worldwide concern. The present study focused on isolation, molecular identification, morphological and biochemical characterization of bacteria possessing hydrocarbon- degrading properties. The study also aimed at optimizing appropriate culture conditions for the isolates as well as screening for alkane hydroxylase enzyme. Out of twenty one microbes isolated, nine were selected based on their ability to utilize different hydrocarbons. The isolates were observed to mineralize heating oil, hexane, octane, toluene and diesel oil. PCR amplification of 16S rDNA gene revealed that the isolates belong to six different genera; Pseudomonas, Acinetobacter, Klebsiella, Enterobacter, Salmonella and Ochrobactrum. Based on their ability to degrade diesel oil, three isolates were selected and their growth conditions optimized. Optimum degradation was recorded at less than 1% substrate concentration, pH 7 and temperature range of between 30°C and 37°C and using yeast extract as nitrogen source. GC-MS analyses confirmed their diesel oil degrading properties. Alkane hydroxylase gene from one of the isolates (isolate 1C) was successfully amplified indicating its catabolic capabilities in degrading alkanes. Overall, the characterized isolates may constitute potential candidates for biotechnological application in environmental cleanup of petroleum contaminants.
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    Rotavirus prevalence and seasonal distribution post vaccine introduction in Nairobi county Kenya
    (2023-03-06) Okanya, Patrick; Mbatia, Betty; Gikonyo, Joshua
    Rotaviruses are one of the leading etiological agents of gastroenteritis in young children, for which a monovalent G1P(8) vaccine has been provided for free in Kenyan since July 2014. The main objective was to estimate the post vaccine prevalence and seasonal distribution of rotavirus diarrhea in children less than 5 years in Nairobi County, Kenya. Rotavirus positive samples were collected from children below 5 years of age in two hospitals within Nairobi County where vaccination status was card-confirmed. The children were examined and the demographic and clinical profiles of the children were recorded. Fecal specimens were analyzed for rotavirus antigen using an ELISA kit, followed by characterization by PAGE. Out of the total 323 samples, 49 had detectable rotavirus infection, representing 15.2% prevalence. Age distribution of rotavirus prevalence was as follows: ≤ 6 months-8.5%, 7-12 months-27.4%, 13-24 months - 41.4%, 25-36 months - 16.4% while 36-65 months had 6.3%. Rotavirus diarrhea was more common in wet and cold months of the year, the highest prevalence being observed in August (24.5%), 12.3% in both July and March, while April scored a prevalence of 10.2%. Out of the 49 rotavirus positive children, 48 had vomiting and abdominal cramps while all had fever and watery stool. The prevalence of Rotaviral diarrhea in children less than 5 years in Nairobi County Kenya has greatly reduced following the vaccine introduction and is more common during the wet and cold seasons of the year.
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    Development and evaluation of a loop-mediated isothermal amplification (LAMP) diagnostic test for detection of whipworm, Trichuris trichiura, in faecal samples
    (2023-03-06) Okanya, Patrick; Ngari, M.G.; Mwangi, I.N.
    Whipworm infection or trichuriasis caused by Trichuris trichiura is of major public health concern in sub-Saharan Africa, particularly among pre-school and school-going children. It is among the neglected tropical diseases targeted for elimination through mass drug administration (MDA). One of the outcomes of MDA is a rapid decline in levels of infection intensity, making it difficult to monitor effectiveness of control measures using the conventional Kato–Katz procedure, which relies on the microscopic detection of parasite ova in faecal samples. In the present study, a loop-mediated isothermal amplification (LAMP) test was developed for the detection of T. trichiura infection in faecal samples. LAMP technology offers greater sensitivity and specificity than the microscopy-based tests. A set of four specific primers targeting the internal transcribed spacer 2 region of the ribosomal DNA were designed using Primer Explorer software. DNA was extracted from faecal samples using the alkaline lysis method (HotSHOT) and the LAMP reaction performed at 63°C for 1 h. The amplicons were visualized by both gel electrophoresis and with the naked eye following staining with SYBR green dye. Sensitivity and specificity tests were determined using the standard Kato–Katz diagnostic procedure as a reference test. The developed LAMP assay reliably detected T. trichiura DNA in faecal samples, with a specificity and sensitivity of 88% and 77%, respectively. No cross-reactivity was observed with several common helminth parasites. The developed LAMP assay is an appropriate diagnostic method for the detection of T. trichiura DNA in human faecal samples due to its simplicity, low cost, high sensitivity and specificity.
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    Lanyamycin, a macrolide antibiotic from Sorangium cellulosum, strain Soce 481 (Myxobacteria)
    (2023-03-06) Okanya, Patrick; Mulwa, Lucky; Jansen, Rolf
    Lanyamycin (1/2), a secondary metabolite occurring as two epimers, was isolated from the myxobacterium Sorangium cellulosum, strain Soce 481. The structures of both epimers were elucidated from HRESIMS and 1D and 2D NMR data and the relative configuration of their macrolactone ring was assigned based on NOE and vicinal 1H NMR coupling constants and by calculation of a 3D model. Lanyamycin inhibited HCV infection into mammalian liver cells with an IC50 value of 11.8 µM, and exhibited a moderate cytotoxic activity against the mouse fibroblast cell line L929 and the human nasopharyngeal cell line KB3 with IC50 values of 3.1 and 1.5 μM, respectively, and also suppressed the growth of the Gram-positive bacterium Micrococcus luteus.