Generation of cell suspensions of East African highland bananas through scalps

The improvement of East African highland bananas (EAHBs) using conventional breeding methods is difficult due to their biology and therefore focus on improving them has shifted to exploring methodsfor establishment of embryogenic cell suspensions, which can then be targeted, for genetic transformation. Shoots of four cultivars namely ‘Musakala’, ‘Kibuzi’, ‘Mbwazirume’ and ‘Lwadungu’ were transferred to a multiplication media modified by adding a combination of N6-benzylaminopurine (BAP) and thidiazuron (TDZ) at concentrations of 24.8/0.45, 16.2/1.14, 14.4/3.50, 12.4/4.55, 10/5.68 mM forscalp generation. These media are referred to as M1, M2, M3, M4 and M5, respectively. Two other treatments designated as M6 and M7 with concentrations of 10 mM TDZ and 100 mM BAP, respectivelywere included for comparison purposes. The scalps developed were excised and inoculated into liquid induction medium supplemented with either BAP or Zeatin to generate cell suspensions. Scalpformation was achieved earlier and at much lower concentrations of combined BAP and TDZ than when singly applied. Combinations of 12.4/4.55 and 10/5.68 M BAP/TDZ produced the best scalps. Thestructure of the cell suspension and the rate of cell growth were found to be dependent on the cultivar regardless of the hormone treatment in the induction medium. Cultivars ‘Musakala’, ‘Kibuzi’ and ‘Mbwazime’ produced cell culture of clustered and aggregated cells and high cell numbers, which are a prerequisite for embryo cells development

Mannitol in diuretic resistant nephrotic syndrome: a case report

A 6 year old presented with generalized oedema to Kagando hospital, Uganda, and was diagnosed with nephrotic syndrome. Despite treatment with a fluid restriction, prednisolone and furosemide, the oedema worsened and the weight increased. Mannitol was added to the treatment regime and the oedema improved. The patient was able to be discharged four days later without any diuretic therapy. The case highlights the use of mannitol in the treatment of diuretic resistant oedema with nephrotic syndrome. It provides an alternative therapy for the resource-poor setting to the expensive albumin-furosemide combination often used elsewhere

Social Mobilization and Compliance with Mass Treatment for Lymphatic Filariasis Elimination in Kenya

This study aimed to establish the role of social mobilization in mass drug administration (MDA) uptake during the National Programme to Eliminate Lymphatic Filariasis (LF) in Kenya. MDA for LF based on diethylcarbamazine (DEC) and albendazole using community-based treatment approach has been conducted for three years (2003, 2005 and 2008) in Kwale and Malindi districts. In each district, one high and one low, compliance locations were selected based on 2008 MDA data. From the four locations, nine villages were systematically sampled and a total of 965 randomly selected household heads interviewed. Sixteen focus group discussions with adult and youth male and female groups and separate in-depth interviews with eighty opinion leaders and eighty LF patients with clinical manifestations, purposively selected were conducted. Semi-structured interviews were held separately with fifteen community drug distributors, five health personnel and four LF coordinators also purposively selected. The results showed that knowledge about MDA for LF was not significantly associated with compliance (P>0.05). Seventy three percent in low and 78% in high compliance villages knew about MDA. The most common source of MDA information given by 49% of respondents in high and 40% in low compliance villages were the community drug distributors (CDDs). The content of MDA information received influenced compliance (P< 0.001), 71% in high compared to 61% in low compliance villages received correct information. The frequency of receiving MDA information also influenced compliance (P< 0.001), 65.5% in high compared to 50% in low compliance villages received the correct information at least once before treatment. Opinion towards the source of MDA information was also associated with compliance, 46% in high compared to 43% in low compliance villages considered the source as adequate (P< 0.001). The study results show that for MDA to be successful, information dissemination should be done by all stakeholders with the health personnel taking the lead role so that more adequate and factual content is relayed. Community sensitization and mobilization should be done repeatedly for all to get the information in good time to comply with treatment. Keywords: Compliance; Lymphatic Filariasis; Mass Drug Administration; Social Mobilizatio

In vitro embryo rescue and plant regeneration following self-pollination with irradiated pollen in cassava (Manihot esculenta Crantz)

Cassava is a highly heterozygous species; hence, current methods used in classical cassava breedingcannot match the urgent need to high yielding varieties. Recently, progress was made through androgenesis and gynogenesis as pathways for raising doubled cassava haploid lines to overcome problems associated with cassava’s inherent reproductive biology, but these efforts were limited (nocandidate cassava plantlets were regenerated). For the first time, this study shows that pollen irradiation coupled with self-pollination and embryo rescue regenerated 62 candidate cassava plantlets. Plants of an elite cassava variety, Nase14, served as a mother plant and as the pollen donor for the irradiation. Irradiation dosages of 50 to 250 Gray studied across five pollination events and 300 or 500 Gray in one pollination event caused a reduction in pollen germination up to 67.0%. By 15 days after pollination (DAP) with irradiated pollen, up to 89.7% of the pollinated flowers had aborted. By embryo rescue time (42 DAP), significant differences were observed in number of fruits, seeds and embryos generated, with the non-irradiated pollen treatments having significantly higher numbers. Sixteen (16) heterozygous SSR markers in the parent and ploidy analysis showed that none of the regenerated plants was haploid or homozygous. However, the plantlets resulting from pollination with non-irradiated pollen had 56.2% homozygous loci, while progeny derived from irradiated treatments had frequencies of homozygous loci between 28.1 and 55.0%. This is the first time to use irradiated pollen in cassava as a pathway to generate candidate plantlets as an initial step in double haploid production.Key words: Cassava, doubled haploids, embryo rescue, plant regeneration, pollen germination, pollenirradiation

Phenotypic and genotypic analyses to guide selection of reverse transcriptase inhibitors in second-line HIV therapy following extended virological failure in Uganda

Objectives We investigated phenotypic and genotypic resistance after 2 years of first-line therapy with two HIV treatment regimens in the absence of virological monitoring. Methods NORA [Nevirapine OR Abacavir study, a sub-study of the Development of AntiRetroviral Therapy in Africa (DART) trial] randomized 600 symptomatic HIV-infected Ugandan adults (CD4 cell count <200 cells/mm3) to receive zidovudine/lamivudine plus abacavir (cABC arm) or nevirapine (cNVP arm). All virological tests were performed retrospectively, including resistance tests on week 96 plasma samples with HIV RNA levels ≥1000 copies/mL. Phenotypic resistance was expressed as fold-change in IC50 (FC) relative to wild-type virus. Results HIV-1 RNA viral load ≥1000 copies/mL at week 96 was seen in 58/204 (28.4%) cABC participants and 21/159 (13.2%) cNVP participants. Resistance results were available in 35 cABC and 17 cNVP participants; 31 (89%) cABC and 16 (94%) cNVP isolates had a week 96 FC below the biological cut-off for tenofovir (2.2). In the cNVP arm, 16/17 participants had resistance mutations synonymous with high-level resistance to nevirapine and efavirenz; FC values for etravirine were above the biological cut-off in 9 (53%) isolates. In multivariate regression models, K65R, Y115F and the presence of thymidine analogue-associated mutations were associated with increased susceptibility to etravirine in the cABC arm. Conclusions Our data support the use of tenofovir following failure of a first-line zidovudine-containing regimen and shed further light on non-nucleoside reverse transcriptase inhibitor hypersusceptibility

Integrated Mapping of Neglected Tropical Diseases: Epidemiological Findings and Control Implications for Northern Bahr-el-Ghazal State, Southern Sudan

Integrated control of neglected tropical diseases (NTDs) is being scaled up in a number of developing countries, because it is thought to be more cost-effective than stand-alone control programmes. Under this approach, treatments for onchocerciasis, lymphatic filariasis (LF), schistosomiasis, soil-transmitted helminth (STH) infection, and trachoma are administered through the same delivery structure and at about the same time. A pre-requisite for implementation of integrated NTD control is information on where the targeted diseases are endemic and to what extent they overlap. This information is generated through surveys that can be labour-intensive and expensive. In Southern Sudan, all of the above diseases except onchocerciasis require further mapping before a comprehensive integrated NTD control programme can be implemented. To determine where treatment for which disease is required, integrated surveys were conducted for schistosomiasis, STH infection, LF, and loiasis, throughout one of ten states of the country. Our results show that treatment is only required for urinary schistosomiasis and STH in a few, yet separate, geographical area. This illustrates the importance of investing in disease mapping to minimize overall programme costs by being able to target interventions. Integration of survey methodologies for the above disease was practical and efficient, and minimized the effort required to collect these data