Bismark Sarfo

Cerebrospinal fluid and serum biomarkers of cerebral malaria mortality in Ghanaian children

BackgroundPlasmodium falciparum can cause a diffuse encephalopathy known as cerebral malaria (CM), a major contributor to malaria associated mortality. Despite treatment, mortality due to CM can be as high as 30% while 10% of survivors of the disease may experience short- and long-term neurological complications. The pathogenesis of CM and other forms of severe malaria is multi-factorial and appear to involve cytokine and chemokine homeostasis, inflammation and vascular injury/repair. Identification of prognostic markers that can predict CM severity will enable development of better intervention.MethodsPostmortem serum and cerebrospinal fluid (CSF) samples were obtained within 2–4 hours of death in Ghanaian children dying of CM, severe malarial anemia (SMA), and non-malarial (NM) causes. Serum and CSF levels of 36 different biomarkers (IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, Eotaxin, FGF basic protein, CRP, G-CSF, GM-CSF, IFN-γ, TNF-α, IP-10, MCP-1 (MCAF), MIP-1α, MIP-1β, RANTES, SDF-1α, CXCL11 (I-TAC), Fas-ligand [Fas-L], soluble Fas [sFas], sTNF-R1 (p55), sTNF-R2 (p75), MMP-9, TGF-β1, PDGF bb and VEGF) were measured and the results compared between the 3 groups.ResultsAfter Bonferroni adjustment for other biomarkers, IP-10 was the only serum biomarker independently associated with CM mortality when compared to SMA and NM deaths. Eight CSF biomarkers (IL-1ra, IL-8, IP-10, PDGFbb, MIP-1β, Fas-L, sTNF-R1, and sTNF-R2) were significantly elevated in CM mortality group when compared to SMA and NM deaths. Additionally, CSF IP-10/PDGFbb median ratio was statistically significantly higher in the CM group compared to SMA and NM groups.ConclusionThe parasite-induced local cerebral dysregulation in the production of IP-10, 1L-8, MIP-1β, PDGFbb, IL-1ra, Fas-L, sTNF-R1, and sTNF-R2 may be involved in CM neuropathology, and their immunoassay may have potential utility in predicting mortality in CM.

Proinflammatory Cytokines and the Hypermetabolism of Children with Sickle Cell Disease

Sickle cell anemia (HbSS) includes chronic inflammation, but the origin is unclear. We hypothesized that in stable HbSS patients the inflammation was associated with hypermetabolism. We compared selected hypermetabolic and key Immuno-modulator indicators in HbSS versus control children and examined associations between measures of hypermetabolism and inflammation. Twelve fasting asymptomatic HbSS children 6–12 years and 9 controls matched for age, gender and fat mass (FM) were studied. Proportional reticulocyte count (retic%) and resting energy expenditure (REE) represented hypermetabolism, and C-reactive protein (CRP) Indicated Inflammation. Proinflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), chemokine monocyte chemoattractant proteln-1 (MCP-1), and energy balance cytokine leptin were measured. Methods were indirect calorimetry, enzyme-linked immunosorbent assay, and radioimmunoassay. Statistical analysis included simple correlation and regression analysis. REE (51 ± 6 vs. 43 ± 12 kcal/kg per fat-free mass (FFM), mean ± SD), retic% (12 ± 4 vs. 0.7 ± 0.3%), CRP (5 ± 3 vs. 0.3 ± 0.4 mg/liter), and IL-6 (71 ± 40 vs. 20 ± 7 pg/ml) were significantly higher for HbSS than controls (P < 0.05). Conversely, leptin (0.1 ± 0.1 vs. 2 ± 1 µg/liter per kgFM) and MCP-1 (34 ± 5 vs. 41 ± 4 pg/ml) were significantly lower for the HbSS subjects (P < 0.01). TNF-α was not significantly different. There were no significant associations between REE or retic% and any cytokine measured. However, CRP was significantly associated with REE in HbSS (r = 0.8, P = 0.003) and an important predictor of REE/FFM. We provide new evidence for low circulating levels of inflammatory chemokine MCP-1 in stable HbSS children, confirm mostly low cytokine levels, inflammation, and hypermetabolism and demonstrate association of hypermetabolism with inflammation via CRP but not via cytokines.

Plasmodium berghei ANKA infection increases Foxp3, IL-10 and IL-2 in CXCL-10 deficient C57BL/6 mice

BackgroundCerebral malaria (CM) is a major cause of malaria mortality. Sequestration of infected red blood cells and leukocytes in brain vessels coupled with the production of pro-inflammatory factors contribute to CM. CXCL-10 a chemokine that is chemotactic to T cells has been linked to fatal CM. Mice deficient for CXCL-10 gene are resistant to murine CM, while antibody ablation of CXCL-10 enhanced the production of regulatory T cells (CD4+Cd25+Foxp3+) and IL-10 which regulate the immune system. Interleukin-2 (IL-2), a pro-inflammatory cytokine implicated in malaria pathogenesis has also been shown to be a key regulator of Foxp3. However the role of Foxp3 in resistant murine CM is not well understood.MethodsThe hypothesis that resistance of CXCL-10-/- mice to murine CM may be due to enhanced expression of Foxp3 in concert with IL-10 and IL-2 was tested. CXCL-10-/- and WT C57BL/6 mice were infected with Plasmodium berghei ANKA and evaluated for CM symptoms. Brain, peripheral blood mononuclear cells (PBMCs) and plasma were harvested from infected and uninfected mice at days 2, 4 and 8. Regulatory T cells (CD4+CD25+) and non-T regs (CD4+CD25-) were isolated from PBMCs and cultured with P. berghei antigens in vitro with dendritic cells as antigen presenting cells. Regulatory T cell transcription and specific factor Foxp3, was evaluated in mouse brain and PBMCs by realtime-PCR and Western blots while IL-10, and IL-2 were evaluated in plasma and cultured supernatants by ELISA.ResultsWild type mice exhibited severe murine CM symptoms compared with CXCL-10-/- mice. Foxp3 mRNA and protein in brain and PBMCs of CXCL-10-/- mice was significantly up-regulated (p < 0.05) by day 4 post-infection (p.i) compared with WT. Plasma levels of IL-10 and IL-2 in infected CXCL-10-/- were higher than in WT mice (p < 0.05) at days 2 and 4 p.i. Ex-vivo CD4+CD25+ T cells from CXCL-10-/- re-stimulated with P. berghei antigens produced more IL-10 than WT CD4+CD25+ T cells.ConclusionThe results indicate that in the absence of CXCL-10, the resulting up-regulation of Foxp3, IL-10 and IL-2 may be involved in attenuating fatal murine CM.

Identification of surface-membrane P-type ATPases resembling fungal K+- and Na+-ATPases, in Trypanosoma brucei, Trypanosoma cruzi and Leishmania donovani

Abstract Genomic DNA fragments encoding nine, novel, P-type ATPases in trypanosomatid organisms were amplified in PCR, using degenerate oligonucleotide primers that recognize the ATP-binding and -phosphorylation sites present in all P-type ATPases. Subsequent phylogenetic analysis, based on the presence of conserved motifs in predicted peptide sequences for six Trypanosoma brucei, T. cruzi or Leishmania donovani PCR fragments, identified calcium-, proton- and phospholipid-translocating ATPases. DNA fragments that predict proteins homologous to the fungal, type-IID, P-type, ATPase pumps that transport Na+ or K+ ions were also present in T. brucei (TBCA1; 1022 nucleotides representing 340 amino acids), T. cruzi (TCNA1; 1022 nucleotides representing 340 amino acids) and L. donovani (LDCA1; 1031 nucleotides representing 343 amino acids). Southern blots showed that the Na+-ATPases were each present as a single-copy gene. The LDCA1 fragment was used to clone the complete LDCA1 gene from an L. donovani genomic-DNA library. The LDCA1 gene encodes a protein, of 1047 amino acids, with a predicted molecular mass of 115,501 Da. The results of analyses based on northern blots and the rapid amplification of cDNA ends (RACE) indicated that LDCA1 was expressed in promastigotes and amastigotes from axenic cultures and in animal-derived amastigotes. TBCA1 was expressed, as a 5.0-kb transcript, in procyclic culture stages and bloodstream trypomastigotes, with the 5.0-kb message up-regulated six-fold in the trypomastigote stage. Western blots probed with an antibody to the partial TBCA1 peptide identified a 150-kDa protein that was detected, by immunofluorescence, on the surface membrane of procyclic T. brucei.

HIV-related knowledge, perceptions, attitudes, and utilisation of HIV counselling and testing: a venue-based intercept commuter population survey in the inner city of Johannesburg, South Africa

Background HIV counselling and testing (HCT) and knowledge about HIV have been key strategies utilised in the prevention and control of HIV/AIDS worldwide. HIV knowledge and uptake of HCT services in sub-Saharan Africa are still low. This study was conducted to determine factors associated with HCT and HIV/AIDS knowledge levels among a commuter population in Johannesburg, South Africa. Objective To identify the factors associated with HCT uptake among the commuter population. Design A simple random sampling method was used to select participants in a venue-based intercept survey at a taxi rank in the Johannesburg Central Business District. Data were collected using an electronic questionnaire. Logistic regression analysis assessed factors associated with HIV testing stratified by gender. Results 1,146 respondents were interviewed, the maority (n=579, 50.5%) were females and (n=780, 68.1%) were over 25 years of age. Overall HCT knowledge was high (n=951, 83%) with more females utilising HCT facilities. There was a significant difference in HIV testing for respondents living closer to and further away from health facilities. Slightly more than half of the respondents indicated stigma as one of the barriers for testing (n=594, 52%, p-value=0.001). For males, living with a partner (aOR: 1.68, 95% CI: 1.02–2.78, p-value: 0.041) and possessing a post-primary education were positively associated with testing (aOR: 2.00, 95% CI: 1.15–3.47, p-value: 0.014), whereas stigma and discrimination reduced the likelihood of testing (aOR: 0.40, 95% CI: 0.31–0.62, p-value: <0.001). For females, having one sexual partner (aOR: 2.65, 95% CI: 1.19–5.90, p-value: 0.017) and a low perceived benefit for HIV testing (aOR: 0.54, 95% CI: 0.30–0.96, p-value: 0.035) were associated with HIV testing. Conclusion The overall HIV/AIDS knowledge was generally high. Gender-specific health education and HIV intervention programmes are needed for improved access to HCT services. One favourable intervention would be the use of home-based HCT programmes.

Factors associated with syphilis screening uptake among pregnant women in health facilities in Brong Ahafo Region of Ghana

BackgroundCongenital syphilis is a consequence of undiagnosed, untreated, or inadequately treated maternal syphilis and results in serious adverse outcomes. It is easily diagnosed and in Ghana it is treated at points of care free; yet most pregnant women attending antenatal clinic (ANC) in Ghana are not screened. This study identified some factors influencing syphilis screening uptake at medical facilities during pregnancy in the Brong Ahafo Region of Ghana.MethodA cross-sectional study was conducted in two districts in Brong Ahafo Region of Ghana. All the health facilities in the selected districts that run antenatal services were assessed on their preparedness to screen syphilis for pregnant women. Interviews were conducted among 390 pregnant women attending ANC at five hospitals in the two districts to identify individual and community level barriers to syphilis screening.ResultsIn all 37 health facilities conducted antennal clinics in the two districts in 2013, 75.7% of the health facilities were public; Techiman had the higher number of health facilities (64.9%), test kits were available in only 29.7% of the health facilities and 43.2% of 37 health facilities were conducting syphilis screening. Majority of the pregnant women (57.7%) were within the age range of 20–29 years, 53.1% were in their third trimester, 32.6% in second trimester and 14.3% were in the first trimester. Syphilis screening was 52.4% among 37 health facilities. Among 390 pregnant women who participated in the study syphilis screening was 21.1%. At the health facility level, screening was significantly associated with the type of hospital (whether private or public), availability of test kits, and trained personnel, such as doctors/midwives and syphilis education. At the individual level, attending a public hospital (OR=5.49; 95% CI=1.71-17.65), willingness to request screening (OR=2.72; 95% CI=1.09-5.88), and being in the third trimester of pregnancy (OR=16.47; 95% CI=2.02-132.81) were significantly associated with syphilis screening uptake.ConclusionDespite government’s free screening policy for syphilis among pregnant women, the coverage of antenatal screening is still low. Training of lower level health workers and regular supply of logistics are crucial for the success of the syphilis prevention programme.

HIV Case Management Support Service Is Associated with Improved CD4 Counts of Patients Receiving Care at the Antiretroviral Clinic of Pantang Hospital, Ghana

Background Factors associated with individual patient-level management of HIV have received minimal attention in sub-Saharan Africa. This study determined the association between support services and cluster of differentiation 4 (CD4) counts among HIV patients attending ART clinic in Ghana. Methodology This was a cross-sectional study involving adults with HIV recruited between 1 August 2014 and 31 January 2015. Data on support services were obtained through a closed-ended personal interview while the CD4 counts data were collected from their medical records. Data were entered into EpiData and analyzed using Stata software. Results Of the 201 patients who participated in the study, 67% (129/191) received case management support service. Counseling about how to prevent the spread of HIV (crude odds ratio (cOR) (95% confidence interval (CI)) (2.79 (1.17–6.68)), mental health services (0.2 (0.04–1.00)), and case management support service (2.80 (1.34–5.82))) was associated with improved CD4 counts of 350 cells/mm3 or more. After adjusting for counseling about how to prevent the spread of HIV and mental health services, case management support service was significantly associated with CD4 counts of 350 cells/mm3 or more (aOR = 2.36 (CI = 1.01–5.49)). Conclusion Case management support service for HIV patients receiving ART improves their CD4 counts above 350 cells/mm3. Incorporating HIV case management services in ART regimen should be a priority in sub-Saharan Africa.

Potential barriers to rapid testing for human immunodeficiency virus among a commuter population in Johannesburg, South Africa.

Background This study aimed to determine barriers to accessing human immunodeficiency virus (HIV) counseling and testing (HCT) services among a commuter population. Methods A cross-sectional, venue-based intercept survey was conducted. Participants were recruited during a 2-day community campaign at the Noord Street taxi rank in Johannesburg, South Africa. Data were collected using a self-administered questionnaire loaded onto an electronic data collection system and analyzed using Stata software. Factors contributing to barriers for HCT were modeled using multivariate logistic regression. Results A total of 1,146 (567 male and 579 female) individuals were interviewed; of these, 51.4% were females. The majority (59.5%) were aged 25–35 years. Significant factors were age group (15–19 years), marital status (married), educational level (high school), distance to the nearest clinic (>30 km), area of employment/residence (outside inner city), and number of sexual partners (more than one). Participants aged 15–19 years were more likely to report low-risk perception of HIV as a barrier to HCT (odds ratio [OR] 1.62; 95% confidence interval [CI] 1.01–2.59), the married were more likely to report low-risk perception of HIV as a barrier to HCT (OR 1.49; 95% CI 1.13–1.96), and those living outside the inner city were more likely to report lack of partner support as a potential barrier (OR 1.94; 95% CI 1.34–2.80), while those with a high school education were more likely to report poor health worker attitude as a potential barrier to HIV testing (OR 2.17; 95% CI 1.36–3.45). Conclusion Age, marital status, occupation, educational level, area of employment and residence, distance to the nearest clinic, and number of sexual partners were factors significantly associated with barriers to HIV testing in the study population. Future HIV intervention targeting this population need to be reinforced in order to enhance HIV testing while taking cognizance of these factors.