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Isolation and Characterization of SARS-CoV-2 in Kenya

American Journal of Molecular Biology 2024年第2期14卷 66-83页

作者： Albina Makio Robinson Mugasiali Irekwa Matthew Mutinda Munyao Caroline Wangui Njoroge Peter Kipkemboi Rotich tonny teya nyandwaro Joanne Jepkemei Yego Anne Wanjiru Mwangi James Hungo Kimotho Ronald Tanui Vincent Rutto Samson Muuo Nzou Center for Traditional Medicine and Drug Research Kenya Medical Research Institute Nairobi Kenya Center for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Department of Biochemistry and Microbiology Rhodes University Grahamstown South Africa Center for Virus Research Kenya Medical Research Institute Nairobi Kenya

The discovery of Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) in Wuhan, Hubei province, China, in December 2019 raised global health warnings. Quickly, in 2020, the virus crossed borders and infected individuals across the world, evolving into the COVID-19 pandemic. Notably, early signs of the virus’s existence were observed in various countries before the initial outbreak in Wuhan. As of 12th of April, the respiratory disease had infected over 762 million people worldwide, with over 6.8 million deaths recorded. This has led scientists to focus their efforts on understanding the virus to develop effective means to diagnose, treat, prevent, and control this pandemic. One of the areas of focus is the isolation of this virus, which plays a crucial role in understanding the viral dynamics in the laboratory. In this study, we report the isolation and detection of locally circulating SARS-CoV-2 in Kenya. The isolates were cultured on Vero Cercopithecus cell line (CCL-81) cells, RNA extraction was conducted from the supernatants, and reverse transcriptase-polymerase chain reaction (RT-PCR). Genome sequencing was done to profile the strains phylogenetically and identify novel and previously reported mutations. Vero CCL-81 cells were able to support the growth of SARS-CoV-2 in vitro, and mutations were detected from the two isolates sequenced (001 and 002). Genome sequencing revealed the circulation of two isolates that share a close relationship with the Benin isolate with the D614G common mutation identified along the S protein. These virus isolates will be expanded and made available to the Kenya Ministry of Health and other research institutions to advance SARS-CoV-2 research in Kenya and the region.

关键词： SARS-CoV-2 COVID-19 Whole Genome Sequencing Phylogenetic Analysis Nucleotide Substitutions Amino Acid Changes

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Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya

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American Journal of Molecular Biology 2022年第2期12卷 43-53页

作者： Otilmoi Poul Stephen tonny teya nyandwaro Robinson Mugasiali Irekwa Rebecca Wanjiku Waihenya Matthew Mutinda Munyao Peter Kipkemboi Rotich Caroline Wangui Njoroge Anne Wanjiru Mwangi Joanne Jepkemei Yego Nicole Sian Tanchu Dawala Koromtili Oumar Grace Ngendo Kanyita Primrose Muthoni Ndungu Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Innovation Technology Transfer Department Kenya Medical Research Institute (ITTD-KEMRI) Nairobi Kenya Zoology Department School of Biological Sciences College of Pure and Applied Sciences Jomo Kenyatta University of Agriculture and Technology Nairobi Kenya Production Department KEMRI Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine Kenya Medical Research Institute (NUITM-KEMRI) Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya

Background: A marked decrease in malaria-related deaths worldwide has been attributed to the administration of effective antimalarials against Plasmodium falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller gene (Pfk13) has been reported to develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) involved in chloroquine (CQ) resistance. Method: In this study, archived 280 samples were collected from Alupe primary school children in Busia, Western Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive giving a prevalence rate of 24%. The sixty-seven were subjected to PCR amplification for the molecular marker resistance to Pfcrt. After PCR amplification, the amplicons were purified and sequenced using Sanger Sequencing. The sequence data were analyzed using BioEdit software to identify point mutations. Results: 14 samples sequences were analyzed on Bioedit software giving the following amino acid changes F76C, Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F). New mutations have been reported at position 76 leading to an amino acid change, one of Pfcrt gold standard biomarkers. However, amino acid changes Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F are newly reported giving an increase in Pfcrt prevalence of concern from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as shown below. Generally, the results showed a continuous resistance of *** to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene pre

关键词： P. falciparum Chloroquine Drug Resistance Malaria P. falciparum Chloroquine Resistance Transporter Gene Mutations

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Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis

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American Journal of Molecular Biology 2023年第2期13卷 127-139页

作者： Dawala Koromtili Oumar Matthew Mutinda Munyao Anne Wanjiru Mwangi Rebecca Wanjiku Waihenya Peter Kipkemboi Rotich Robinson Mugasiali Irekwa tonny teya nyandwaro Caroline Wangui Njoroge Joanne Jepkemei Yego Primrose Muthoni Ndungu Sharlene Kerubo Mageto Damaris Mutethya Kilei Otilmoi Poul Stephen Nicole Sian Tanchu Grace Ngendo Kanyita Shingo Inoue Lucy Gitau Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine-Kenya Medical Research Institute Project Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Zoology Department Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya Department of Microbiology Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Center for Biotechnology Research and Development Kenya Medical Research Institute Nairobi Kenya

Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of the primary phase are not specific, the existing diagnostic methods have low sensitivity and specificity, and the available drugs have some toxicity. New, robust, and cost-effective techniques are needed for the early identification of parasites. This study aimed to assess the sensitivity and specificity of two different types of polyclonal antibodies against T. b. gambiense using antigen detection ELISA. Polyclonal antibodies against the expressed proteins Tbg I2 and Tbg I17 were produced using New Zealand white rabbits. The antibody titer measured was greater than 32 g/L after the 3rd immunization for the expressed protein Tbg I2. For the expressed protein Tbg I17, the antibody titer measured was greater than 32 g/L after the 4th immunization. The sensitivity and specificity of the Tbg I2 polyclonal antibody confirmed with Polymerase Chain Reaction (PCR) as gold standard were respectively 89.5% and 80.6%, while for the Tbg I17 polyclonal antibody, the sensitivity and specificity were respectively 92.1% and 88.9%. The area under the curve for the Tbg I2 polyclonal antibody was 0.90 ± 0.032, while for the Tbg I17 polyclonal antibody, the area under the curve was 0.92 ± 0.0. The Tbg I17 polyclonal antibody produced in New Zealand white rabbits has good sensitivity and good specificity;it can be successfully used in the diagnosis of HAT.

关键词： Human African Trypanosomiasis Polyclonal Antibody Tbg I2 Expressed Protein Tbg I17 Expressed Protein Sensitivity Specificity

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Assessing adaptability and response of vegetation to glacier recession in the afro-alpine moorland terrestrial ecosystem of Rwenzori Mountains

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Journal of Mountain Science 2016年第9期13卷 1584-1597页

作者： tonny J.OYANA Bob R.NAKILEZA Spatial Analytics and Informatics Core Department of Preventive Medicine The University of Tennessee Health Science Center Mountain Resource Centre Department of Geography Geo-informatics and Climate Sciences Makerere University

The objective of this study was to explore vegetation adaptability in a changing afro-alpine moorland terrestrial ecosystem on Mt. Rwenzori and to determine whether there were any links with response of vegetation to glacier recession. We analyzed the composition and distribution of plant species in relation to soils, geomorphic processes, and landscape positions in the Alpine zone. To accomplish this objective, archival data sources and published reports for this ecosystem were reviewed. A field trip was conducted in 2010 to study in detail seven vegetation sampling plots that were systematically selected using GIS maps and a nested-quadrat sampling design framework along an altitudinal gradient in the lower and upper alpine zones. Using these sampling plots, 105 vegetation and 13 soil samples were assessed in the alpine zone. Soil samples were taken for laboratory testing and analysis. The results show statistically significant differences in pH, OM, N, P, Ca, Mg, and K pools between soils samples drawn from the lower and upper alpine sites （p 〈 0.0033）. Furthermore, we observed a significant vegetation formation with numerous structural forms, but there was a limited diversity of speeies. The most significant forms included Alchemilla carpets, Bogs, Dendrosenecio woodland, and Scree slopes. The lower alpine area （3500-3900 masl） had a more diverse plant species than other areas, especially Alchemilla argyrophylla and Dendrosenecio adnivalis species that were evident due to well-drained deeper soils. The Alchemilla subnivalis were evident at a higher altitude of above 4000 mask Shifts in the Astareeeae （e.g. Senecio species） were particularly prominent even on recently deglaciated areas. The spatial variations of species distribution, structure, and composition suggest there are serious implications in terms of ecosystem adaptability, resilience, and stability that require further evaluation.

关键词： Alpine Plant species Species abundance Tropical afro-alpine ecosystems Afro-alpinemoorland

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Comparative short-term effects of sewage sludge and its biochar on soil properties, maize growth and uptake of nutrients on a tropical clay soil in Zimbabwe

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Journal of Integrative Agriculture 2016年第6期15卷 1395-1406页

作者： Willis Gwenzi Moreblessing Muzava Farai Mapanda tonny P Tauro Department of Soil Science and Agricultural Engineering Faculty of Agriculture University of Zimbabwe Marondera College of Agricultural Science and Technology University of Zimbabwe

Soil application of biochar from sewage could potentially enhance carbon sequestration and close urban nutrient balances. In sub-Saharan Africa, comparative studies investigating plant growth effect and nutrients uptake on tropical soils amended with sewage sludge and its biochar are very limited. A pot experiment was conducted to investigate the effects of sewage sludge and its biochar on soil chemical properties, maize nutrient and heavy metal uptake, growth and biomass partitioning on a tropical clayey soil. The study compared three organic amendments; sewage sludge （SS）, sludge biochar （SB） and their combination （SS＋SB） to the unamended control and inorganic fertilizers. Organic amendments were applied at a rate of 15 t ha-1 for SS and SB, and 7.5 t ha-1 each for SS and SB. Maize growth, biomass production and nutrient uptake were significantly improved in biochar and sewage sludge amendments compared to the unamended control. Comparable results were observed with F, SS and SS＋SB on maize growth at 49 d of sowing. Maize growth for SB, SS, SS＋SB and F increased by 42, 53, 47, and 49%, respectively compared to the unamended control. Total biomass for SB, SS, SS＋SB, and F increased by 270,428, 329, and 429%, respectively compared with the unamended control. Biochar amendments reduced Pb, Cu and Zn uptakes by about 22% compared with sludge alone treatment in maize plants. However, there is need for future research based on the current pot experiment to determine whether the same results can be produced under field conditions.

关键词： maize growth nutrient uptake sludge biochar soil chemical properties

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Extraction,quantification and characterization of oil from pumpkin seeds

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International Journal of Agricultural and Biological Engineering 2015年第1期8卷 98-102页

作者： tonny Kukeera Noble Banadda Peter Tumutegyereize Nicholas Kiggundu Ratibu Asuman Department of Agricultural&Bio-Systems Engineering Makerere UniversityKampalaUganda Chemistry Laboratory Uganda Industrial Research InstituteKampalaUganda

A quantification and characterization study of oil from pumpkin seeds was carried out on three pumpkin varieties,namely,Japanese type of the Curcubita maxima species,Green Kabocha and butternut squash of the Cucurbita moschanta *** extraction was done using the Soxhlet method with petroleum ether as the *** physicochemical properties of acid value,iodine value,specific gravity and refractive index were *** results were analyzed statistically using one way ANOVA at a 5% level of *** mean values of oil content,acid value,iodine value,specific gravity and refractive index for Japanese type pumpkin were found to be 35.67%,2.24 mg KOH/g,26.45 mg I2/100 g,0.9126 and 1.47,*** Green Kabocha,the mean values of the above five properties were 30.12%,3.35 mg KOH/g,25.3 mg I2/100 g,0.9126 and 1.469,*** the Butternut squash type the mean values of the above five properties were 34.76%,3.74 mg KOH/g,26.61 mg I2,0.9131 and 1.471,*** can be noted that from the low acid values of the pumpkin oil,pumpkin oil can be identified as *** the low iodine value indicated that the pumpkin oil was *** addition,the low iodine value also suggested that the oil contains few unsaturated bonds and therefore has low susceptibility to oxidative *** a comparison of extraction rates from other oil sources in Uganda such as cotton(15%-20%),soybeans(19%-21%),the obtained results in this study suggested that pumpkin seeds are a viable source of vegetable oils that can be utilized for commercial vegetable oil extraction in Uganda.

关键词： pumpkin seeds pumpkin varieties oil extraction physicochemical properties vegetable oil characterization

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Efficient photoelectrochemical conversion of CO_(2)to syngas by photocathode engineering

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Green Energy & Environment 2022年第3期7卷 545-553页

作者： Sheng Chu Pengfei Ou Roksana tonny Rashid Yuyang Pan Daolun Liang Huiyan Zhang Jun Song Key Laboratory of Energy Thermal Conversion and Control of Ministry of Education School of Energy and EnvironmentSoutheast UniversityNanjing210096China Department of Electrical and Computer Engineering McGill UniversityMontrealQC H3A 0E9Canada Department of Mining and Materials Engineering McGill UniversityMontrealQC H3A 0C5Canada

The synthesis of renewable chemical fuels from CO_(2) and H_(2)O via photoelectrochemical(PEC)route reprensents a promising room-temperature approach for transforming greenhouse gas into value-added chemicals(e.g.,syngas),but to date it has been hampered by the lack of efficient photocathode for CO_(2) ***,we report efficient PEC CO_(2) reduction into syngas by photocathode *** photocathode is consisting of a planar p-n Si junction for strong light harvesting,GaN nanowires for efficient electron extraction and transfer,and Au/TiO_(2)for rapid electrocatalytic syngas *** photocathode yields a record-high solar energy conversion efficiency of 2.3%.Furthermore,desirable syngas compositions with CO/H_(2)ratios such as 1:2 and 1:1 can be produced by simply varying the size of Au *** calculations reveal that the active sites for CO and H_(2)generation are the facet and undercoordinated sites of Au particles,respectively.

关键词： syngas photocathode photoelectrochemical

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Power hardware in the loop validation of fault ride through of VSC HVDC connected offshore wind power plants

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Journal of Modern Power Systems and Clean Energy 2014年第1期2卷 23-29页

作者： Ranjan SHARMA Qiuwei WU Seung Tae CHA Kim H.JENSEN tonny W.RASMUSSEN JacobØSTEGAARD Siemens Wind Power A/S Borupvej 167330 BranderDenmark Centre for Electric Power and Energy Department of Electrical EngineeringTechnical University of DenmarkElektrovej 3252800 Kongens LyngbyDenmark

This paper presents the power hardware in the loop(PHIL)validation of a feed forward DC voltage control scheme for the fault ride through(FRT)of voltage source converter(VSC)high voltage DC(HVDC)connected offshore wind power plants(WPPs).In the proposed FRT scheme,the WPP collector network AC voltage is actively controlled by considering both the DC voltage error and the AC current from the WPP AC collector system which ensures fast and robust FRT of the VSC HVDC connected offshore *** PHIL tests were carried out in order to verify the efficacy of the proposed feed forward DC voltage control scheme for enhancing the FRT capability of the VSC HVDC connected *** PHIL test results have demonstrated the proper control coordination between the offshore WPP and the WPP side VSC and the efficient FRT of the VSC HVDC connected WPPs.

关键词： Fault ride through High voltage DC(HVDC) Offshore wind power plant Power hardware in the loop(PHIL) Voltage source converter(VSC)

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Pitfalls in an International Development (ID) Project Implementation: A Case Study in a Developing Country

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海外英语 2011年第2X期 272-275页

作者： tonny Kepou School of Management Shanghai University

Literature review noted that donor funded projects and programs are sometimes referred to as International Development: ID projects. ID projects and programs are often being provided for the developing countries like an emerging, developing or least developed country. Such projects and programs are under public sector development specifically designed for economic and social needs of developing countries. These projects or programs are either implemented by recipient governments under a bilateral (or multilateral) agreement with the donor country or through an 'implementing partner' of the donor, frequently a non-governmental organization or professional contractor. The operating environment and culture of the host country also make ID projects different from traditional business projects and make traditional project management tools in the developed world less appropriate (Blunt and Jones, 1992). This sometimes provides big challenges to various project teams. The paper through a Case study approach evaluates an ID Chinese Government funding construction project and its pitfalls through qualitative research approach. Qualitative data was presented and discussed to provide an insight into major challenges in the implementation of ID project of such scale.

关键词： international development (ID) contract documents donor agency

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Audit Committee, Value Creation Efficiency and Capital Employed Efficiency

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Management Studies 2018年第1期6卷 20-29页

作者： Fanny Soewignyo tonny Irianto Soewignyo Universitas Klabat Airmadidi Manado Indonesia

This study contributes a number of innovative and interesting empirical findings with a view of four audit committee characteristics to predict overall value creation efficiency and capital employed efficiency using value added intellectual coefficient （VAICTM） method. Using purposive sampling, 34 property, real estate, and building construction firms listed on Indonesia Stock Exchange in 2011 were selected. Empirical findings could not provide a significant relationship between audit committee characteristics and the overall value creation efficiency as well as capital employed efficiency. This implies that currently, the number of members, number of meetings, number of independent commissioners, and accounting or finance expertise in audit committee cannot be expected as drivers of business value creation in Indonesian context, more specifically for property, real estate, and building construction industry. The overall lack of significant relationships may potentially result from limited human capacity, lack of financial expertise, and inadequate knowledge about the role of audit committee to add value to the business.

关键词： audit committee capital employed efficiency VAICTM value added intellectual coefficient value creation efficiency

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