Neurological & Psychiatric Society of Zambia's Evidence-Based Guidelines for EEG Utilization at the University Teaching Hospital

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Pre-clinical development of BCG.HIVA(CAT), an antibiotic-free selection strain, for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG

In the past, we proposed to develop a heterologous recombinant BCG prime-recombinant modified vaccinia virus Ankara (MVA) boost dual pediatric vaccine platform against transmission of breast milk HIV-1 and Mycobacterium tuberculosis (Mtb). In this study, we assembled an E. coli-mycobacterial shuttle plasmid pJH222.HIVACAT expressing HIV-1 clade A immunogen HIVA. This shuttle vector employs an antibiotic resistance-free mechanism based on Operator-Repressor Titration (ORT) system for plasmid selection and maintenance in E. coli and lysine complementation in mycobacteria. This shuttle plasmid was electroporated into parental lysine auxotroph (safer) strain of BCG to generate vaccine BCG.HIVACAT. All procedures complied with Good Laboratory Practices (GLPs). We demonstrated that the episomal plasmid pJH222.HIVACAT was stable in vivo over a 20-week period, and genetically and phenotypically characterized the BCG.HIVACAT vaccine strain. The BCG.HIVACAT vaccine in combination with MVA.HIVA induced HIV-1- and Mtb-specific interferon γ-producing T-cell responses in newborn and adult BALB/c mice. On the other hand, when adult mice were primed with BCG.HIVACAT and boosted with MVA.HIVA.85A, HIV-1-specific CD8+ T-cells producing IFN-γ, TNF-α, IL-2 and CD107a were induced. To assess the biosafety profile of BCG.HIVACAT-MVA.HIVA regimen, body mass loss of newborn mice was monitored regularly throughout the vaccination experiment and no difference was observed between the vaccinated and naïve groups of animals. Thus, we demonstrated T-cell immunogenicity of a novel, safer, GLP-compatible BCG-vectored vaccine using prototype immunogen HIVA. Second generation immunogens derived from HIV-1 as well as other major pediatric pathogens can be constructed in a similar fashion to prime protective responses soon after birth

Modern Contraceptive and Dual Method Use among HIV-Infected Women in Lusaka, Zambia

HIV-infected women in sub-Saharan Africa are at substantial risk of unintended pregnancy and sexually transmitted infections (STIs). Linkages between HIV and reproductive health services are advocated. We describe implementation of a reproductive health counseling intervention in 16 HIV clinics in Lusaka, Zambia. Between November 2009 and November 2010, 18,407 women on antiretroviral treatment (ART) were counseled. The median age was 34.6 years (interquartile range (IQR): 29.9–39.7), and 60.1% of women were married. The median CD4+ cell count was 394 cells/uL (IQR: 256–558). Of the women counseled, 10,904 (59.2%) reported current modern contraceptive use. Among contraceptive users, only 17.7% reported dual method use. After counseling, 737 of 7,503 women not previously using modern contraception desired family planning referrals, and 61.6% of these women successfully accessed services within 90 days. Unmet contraceptive need remains high among HIV-infected women. Additional efforts are needed to promote reproductive health, particularly dual method use

Comparison of real-time PCR and microscopy for malaria parasite detection in Malawian pregnant women

Abstract Background New diagnostic tools for malaria are required owing to the changing epidemiology of malaria, particularly among pregnant women in sub-Saharan Africa. Real-time PCR assays targeting Plasmodium falciparum lactate dehydrogenase (pfldh) gene may facilitate the identification of a high proportion of pregnant women with a P. falciparum parasitaemia below the threshold of microscopy. These molecular methods will enable further studies on the effects of these submicroscopic infections on maternal health and birth outcomes. Methods The pfldh real-time PCR assay and conventional microscopy were compared for the detection of P. falciparum from dried blood spots and blood smears collected from the peripheral blood of 475 Malawian women at delivery. A cycle threshold (Ct) of the real-time PCR was determined optimizing the sensitivity and specificity of the pfldh PCR assay compared to microscopy. A real-time PCR species-specific assay was applied to identify the contribution to malaria infections of three Plasmodium species (P. falciparum P. ovale and P. malariae) in 44 discordant smear and pfldh PCR assay results. Results Of the 475 women, P. falciparum was detected in 11 (2.3%) by microscopy and in 51 (10.7%) by real-time PCR; compared to microscopy, the sensitivity of real-time PCR was 90.9% and the specificity 91.2%. If a Ct value of 38 was used as a cut-off, specificity improved to 94.6% with no change in sensitivity. The real-time PCR species-specific assay detected P. falciparum alone in all but four samples: two samples were mixed infections with P. falciparum and P. malariae, one was a pure P. malariae infection and one was a pfldh PCR assay-positive/species-specific assay-negative sample. Of three P. malariae infections detected by microscopy, only one was confirmed by the species-specific assay. Conclusions Although microscopy remains the most appropriate method for clinical malaria diagnosis in field settings, molecular diagnostics such as real-time PCR offer a more reliable means to detect malaria parasites, particularly at low levels. Determination of the possible contribution of these submicroscopic infections to poor birth outcomes and maternal health is critical. For future studies to investigate these effects, this pfldh real-time PCR assay offers a reliable detection method

Duration of cART Before Delivery and Low Infant Birthweight Among HIV-Infected Women in Lusaka, Zambia

To estimate the association between duration of combination antiretroviral therapy (cART) during pregnancy and low infant birthweight (LBW), among women ≥37 weeks gestation

Molecular characteristics of Tomato mosaic virus infecting tomato in Uganda

Viral diseases are part of the limiting factors to tomato ( Solanum lycopersicum L.) cultivation worldwide, reducing both the quality and quantity of yield. Tomato mosaic virus (ToMV) is one of the damaging viruses of tomato. This paper describes molecular characteristics of the full length genome of ToMV isolated from tomato in Uganda (ToMV-Ug). The genomic, ribonucleic acid (RNA), of this isolate is 6383 nucleotides (nts) in length, encoding four open reading frames (ORFs). Based on the homology with other ToMV strains, the 5\u2019 proximal 130 kilo dalton (kDa) ORF and its read-through product (180 kDa) are expected to encode two proteins required for viral genome replication; while the 30 kDa middle ORF and the 17.5 kDa 3\u2019 proximal ORF are expected to encode the movement protein (MP) and coat protein (CP), respectively. The 5\u2019- and 3\u2019- untranslated regions (UTRs) are 71 and 201 nts, respectively. Comparison with previously published ToMV sequences showed that ToMV-Ug is 99% identical to ToMV strains from Africa (Egypt and Zimbabwe), as well as diverse locations such as China, Australia, Germany and Japan; suggesting high levels of sequence conservation within this virus. This is the first report detailing molecular analysis of a ToMV isolate from Uganda and the Eastern and Central Africa regions.Les maladies virales font partie des facteurs limitant la production mondiale de la tomate ( Solanum lycopersicum L.), r\ue9duisant \ue0 la fois la quantit\ue9 et la qualit\ue9 du rendement. Le virus de la mosa\uefque de la tomate (ToMV) est l\u2019un des virus endommageant la tomate. Ce papier d\ue9crit les caract\ue9ristiques mol\ue9culaires de la longueur du g\ue9nome de l\u2019isolat ToMV de la tomate en Ouganda (ToMV-Ug). L\u2019acide g\ue9nomique, ribonucl\ue9ique (ARN), de l\u2019isolat a une longueur de 6383 nucl\ue9otides (nts), codant quatre cadres de lecture ouverts (ORFs). Sur la base de l\u2019homologie avec les autres souches de ToMV, le proximal 5\u2019 de 130 kilo dalton (kDa) de l\u2019ORF et sa lecture \ue0 travers le produit (180 kDa) sont esp\ue9r\ue9s coder pour deux prot\ue9ines n\ue9cessaires \ue0 la r\ue9plication du g\ue9nome viral\ua0; alors que les 30 kDa du ORF moyen et les 17,5 kDa du proximal 3\u2019 du ORF sont esp\ue9r\ue9s coder pour le mouvement de la prot\ue9ine (MP) et la prot\ue9ine de l\u2019enveloppe (CP), respectivement. Les r\ue9gions non traduites du 5\u2019 et 3\u2019 (UTRs) sont de 71 et 201 nts, respectivement. La comparaison avec les s\ue9quences (ToMV) pr\ue9c\ue9demment publi\ue9es a montr\ue9 que ToMV-Ug est \ue0 99% identique aux souches ToM de l\u2019Afrique (Egypte et Zimbabw\ue9), ainsi que diverses localit\ue9s telles que la Chine, l\u2019Australie, la Germanie et le Japon\ua0; sugg\ue9rant de hauts niveaux de s\ue9quence de conservation dans ce virus. Ceci est le premier rapport d\ue9taillant l\u2019analyse mol\ue9culaire d\u2019un isolat ToMV d\u2019Ouganda et les r\ue9gions Est et Centre de l\u2019Afrique

Increased prevalence of dhfr and dhps mutants at delivery in Malawian pregnant women receiving intermittent preventive treatment for malaria

In the context of an Intermittent preventive treatment (IPTp) trial for pregnant women in Malawi, P. falci-parum samples from 85 women at enrollment and 35 women at delivery were genotyped for mutations associated with sulfadoxine-pyrimethamine resistance. The prevalence of the highly resistant haplotype with mutations at codons 51 and 108 of dihydrofolate reductase (dhfr) and codons 437 and 540 of dihydropteroate synthase (dhps) increased from 81% at enrollment to 100% at delivery (p=0.01). Pregnant women who were smear-positive at enrollment were more likely to have P. falciparum parasitemia at delivery. These results lend support to concerns that IPTp use may lead to increased drug resistance in pregnant women during pregnancy and emphasize the importance of screening pregnant women for malaria parasites in areas with prevalent SP resistance even when they are already on IPTp

Identifying barriers to ART initiation and adherence: An exploratory qualitative study on PMTCT in Zambia

Background Though antiretroviral therapy (ART) is widely available, HIV positive pregnant women in Zambia are less likely to start and remain on therapy throughout pregnancy and after delivery. This study sought to understand readiness to start ART among HIV pregnant women from the perspectives of both women and men in order to suggest more holistic programs to support women to continue life-long ART after delivery. Methods We conducted a qualitative study with HIV positive pregnant women before and after ART initiation, and men with female partners, to understand readiness to start lifelong ART. We conducted 28 in-depth interviews among women and 2 focus group discussions among male partners. Data were transcribed verbatim and analyzed in NVivo 12 using thematic analysis. Emerging themes from the data were organized using the social ecological framework. Results Men thought of their female partners as young and needing their supervision to initiate and stay on ART. Women agreed that disclosure and partner support were necessary preconditions to ART initiation and adherence and, expressed fear of divorce as a prominent barrier to disclosure. Maternal love and desire to look after one’s children instilled a sense of responsibility among women which motivated them to overcome individual, interpersonal and health system level barriers to initiation and adherence. Women preferred adherence strategies that were discrete, the effectiveness of which, depended on women’s intrinsic motivation. Conclusion The results support current policies in Zambia to encourage male engagement in ART care. To appeal to male partners, messaging on ART should be centered on emphasizing the importance of male involvement to ensure women remain engaged in ART care. Programs aimed at supporting postpartum ART adherence should design messages that appeal to both men’s role in couples’ joint decision-making and women’s maternal love as motivators for adherence