Evaluating the impact of COVID-19 pandemic on complicated malaria admissions and outcomes in the paediatric Ho Teaching Hospital of the Volta Region of Ghana.

Since Ghana recorded its first cases of COVID-19 in early March 2020, healthcare delivery in the country has been hugely affected by the pandemic. Malaria continues to be an important public health problem in terms of morbidity and mortality among children, and it is responsible for significant hospital visits and admission. It is likely that, as with other illnesses, the COVID-19 pandemic may have impacted health seeking behaviour, hospital visits, and admissions of malaria among the paediatric population in Ghana. The aim of this study was to evaluate the impact of COVID-19 pandemic on the admissions and outcome of complicated malaria in the Ho Teaching Hospital of the Volta Region of Ghana. The medical records of children admitted for complicated malaria (cerebral and severe malaria) from 2016 to 2020, were obtained from the admission records of the children. Both demographics and clinical details were collected, and data was analysed using SPSS version 25 statistical software. The yearly differences in the trend and proportions of complicated malaria admissions were performed using rate comparison analysis and Pearson chi-square was used to assess the association between the various demographic factors and yearly admission rates. Clopper-Pearson test statistic was employed to determine the 95% confidence intervals of outcome variables of interest. The year 2020 had the lowest admission for complicated malaria (149, 11.5%; 95% CI: 9.7-13.5) but proportionally had, more cases of cerebral malaria (25, 16.8%; 95% CI: 10.9-24.8), and more deaths (6, 4.0%; 95% CI: 1.5-8.8), compared to the years under review. Children admitted in 2020 had the shortest mean stay on admission (4.34 ±2.48, p<0.001). More studies are needed to further elucidate the impact of the COVID-19 pandemic on the health of children in malaria endemic areas.

Pattern and predictors of immunologic recovery in HIV infected patients.

BACKGROUND: Although anti-retroviral therapy has generally improved the survival of HIV infected patients in many developing countries including Ghana, specific socio-demographic factors could still influence outcome of treatment. This study was designed to identify patient-specific factors that could influence the immune recovery of absolute CD4 count in HIV infected patients. FINDINGS: Hospital records were extracted from two health facilities in Ghana. The impact of socio-demographic factors type of ART and baseline category of CD4 counts were assessed at six monthly interval using robust linear mixed models. RESULTS: A total of 214 follow up records were reviewed at Komfo Anokye Teaching Hospital (KATH) and the Kumasi South Hospital (KSH). One hundred (46.7%) were from KATH and 114 (53.3%) were from KSH. There was a general increase in the level of CD4 counts with time, however this increase significantly slowed down with subsequent reviews (p < 0.001). On the average the rate of CD4 count recovery slowed down by 43.6 cells/µl for every 6 months of follow up (SE = 7.69; p < 0.001). Similarly the recovery of CD4 counts in subjects with an initial high baseline CD4 counts decreased by 192.6 cells/µl (SD error = 42.3, p value ≤0.001). All other variables were not significantly associated with recovery of CD4 counts. CONCLUSION: Our study has demonstrated the well-known phenomenon of CD4 counts increasing after administration of ARTs. CD4 counts increased more rapidly in those with relatively lower initial counts, catching up with those with high CD4 count by 2 years post treatment.

Novel characterization of a breakpoint in F8: an individualized approach to gene analysis when PCR and MLPA results contradict.

Haemophilia A is an X-linked bleeding disorder caused by heterogeneous mutations in the F8 gene. Two inversion hotspots in intron 22 and intron 1, as well as point mutations, small insertions and deletions in the F8 gene account for causal mutations leading to severe haemophilia A. Rarely, novel molecular mechanisms lead to a haemophilia A phenotype which cannot be completely characterized by routine molecular diagnostic methods. Here, we characterized the molecular abnormality in a boy with a severe haemophilia A phenotype. On investigation by PCR and DNA sequencing, exon 18 of F8 repeatedly failed to amplify. However, analysis by multiplex ligation-dependent probe amplification demonstrated the presence of exon 18 sequence, suggesting a more complex rearrangement than a single exon deletion. The analysis of exon 18 and its flanking regions by inverse PCR revealed a complex mutation comprising insertions of extragenic sequences from Xq28 along with a partial duplication of exon 18. Based on the successful analysis and characterization of the familial breakpoint, we developed a PCR-based diagnostic approach to detect this defect in family members in whom no diagnostic test could be offered until this time.

Measuring naturally acquired ex vivo IFN-γ responses to Plasmodium falciparum cell-traversal protein for ookinetes and sporozoites (CelTOS) in Ghanaian adults.

BACKGROUND: A malaria vaccine that targets the sporozoite/liver stage parasites could potentially prevent blood stage infection and the associated clinical symptoms. Identification of sporozoite/liver stage antigens is, therefore, crucial for the development of effective vaccines. Cell-traversal protein for ookinetes and sporozoites (CelTOS) is a highly conserved antigen involved in sporozoite motility and hepatocyte invasion and has been shown to induce significant IFN-γ production in PBMCs from radiation-attenuated sporozoite-immunized malaria-naïve individuals. The aim of this study was to ascertain whether such CelTOS-specific recall responses are also induced in individuals with natural exposure to Plasmodium falciparum. METHODS: Ex vivo IFN-γ responses to 15mer overlapping peptide pools covering the entire sequence of CelTOS and five other candidate antigens, CSP, AMA1, MSP1, TRAP and LSA1, were characterized using PBMCs from 35 malaria exposed adults. Responses to four CelTOS peptide pools (CelTp1, CelTp2, CelTp3 and CelTp4), a pool containing peptides from the entire CelTOS antigen (CelTTp), and pools comprised of overlapping peptides from each of the other five malaria antigens were assessed by ex vivo ELISpot assay. A positive IFN-γ response for stimulants was defined by two criteria; a stimulation index of two or greater relative to the unstimulated control, and a difference of 10 or greater in spot forming cells between stimulant and the unstimulated control. RESULTS: Of the 35 volunteers tested, five had positive IFN-γ recall responses against the four different CelTOS pools while four volunteers made responses against the CelTTp pool; six volunteers were, therefore, positive with CelTOS. By contrast, six volunteers responded to AMA1, seven to LSA1, 15 to MSP1 and two volunteers responded against CSP and TRAP. CONCLUSIONS: These results suggest natural malaria transmission induces CelTOS-specific ex vivo IFN-γ in Ghanaian adults and that the frequency of these responses was similar to those of other previously characterized malaria antigens. These findings support the further evaluation of CelTOS as a pre-erythrocytic candidate antigen for inclusion in a potential multi-antigen vaccine.

Risk factors associated with hepatitis B exposure and the reliability of five rapid kits commonly used for screening blood donors in Ghana.

BACKGROUND: Hepatitis B virus infection (HBV) is one of the most widespread, chronic viral infections in sub-Saharan Africa, and parts of South America. Therefore, efforts are being made to implement strategies aimed at reducing the incidence of hepatitis B viral infections. One route of HBV transmission is through unsafe blood transfusion, which could occur from the use of less sensitive laboratory diagnostic kits. Information on the sensitivity and specificity of these kits is however limited in many developing countries. This study was therefore performed to describe the prevalence of HBV infections and also to evaluate the performance of five rapid immunochromatographic kits commonly used in Ghana. METHODS: A cross-sectional study was designed to describe the prevalence of HBsAg infection and also evaluate the performance of rapid kits used for screening hepatitis B in the northern part of Ghana. RESULTS: A total of 164 prospective blood donors were enrolled in this study from January 2012 to December 2013. The overall true prevalence of HBsAg was 14.6 (95% CI = 9.6 - 21.0). There was no significant association between transmission related factors and HBV infection. The specificities of all five rapid kits were above 97%, however the sensitivities and Youden's indexes were below 60%. A comparison of the reported kit sensitivities to those generated by this study showed significant difference with the study results being lower than the ones reported in the kit literature. CONCLUSION: Our study has shown that rapid HBsAg kits on the Ghanaian markets may not be helpful for screening blood donor samples. We therefore recommend the use of commercially available enzyme linked immunosorbent assays.

Respiratory viruses in children hospitalized for acute lower respiratory tract infection in Ghana.

BACKGROUND: Acute respiratory tract infections are one of the major causes of morbidity and mortality among young children in developing countries. Information on the viral aetiology of acute respiratory infections in developing countries is very limited. The study was done to identify viruses associated with acute lower respiratory tract infection among children less than 5 years. METHOD: Nasopharyngeal samples and blood cultures were collected from children less than 5 years who have been hospitalized for acute lower respiratory tract infection. Viruses and bacteria were identified using Reverse Transcriptase Real-Time Polymerase Chain Reaction and conventional biochemical techniques. RESULTS: Out of 128 patients recruited, 33(25.88%%, 95%CI: 18.5% to 34.2%) were positive for one or more viruses. Respiratory Syncytial Virus (RSV) was detected in 18(14.1%, 95%CI: 8.5% to 21.3%) patients followed by Adenoviruses (AdV) in 13(10.2%, 95%CI: 5.5% to 16.7%), Parainfluenza (PIV type: 1, 2, 3) in 4(3.1%, 95%CI: 0.9% to 7.8%) and influenza B viruses in 1(0.8%, 95%CI: 0.0 to 4.3). Concomitant viral and bacterial co-infection occurred in two patients. There were no detectable significant differences in the clinical signs, symptoms and severity for the various pathogens isolated. A total of 61.1% (22/36) of positive viruses were detected during the rainy season and Respiratory Syncytial Virus was the most predominant. CONCLUSION: The study has demonstrated an important burden of respiratory viruses as major causes of childhood acute respiratory infection in a tertiary health institution in Ghana. The data addresses a need for more studies on viral associated respiratory tract infection.

Mutation analysis in F9 gene of 17 families with haemophilia B from Iran.

Seventeen haemophilia B families from Iran were investigated to determine the causative mutation. All the essential regions of the F9 gene were initially screened by conformational sensitive gel electrophoresis and exons with band shift were sequenced. Seven of the 15 mutations identified in these families were novel mutations. The mutations were authenticated in nine families as other affected members or heterozygous female carriers were available for verification.

Site and type of mutations in the factor VIII gene in patients and carriers of haemophilia A.

Haemophilia A is an X-linked bleeding disorder caused by reduced or absent FVIII (FVIII) protein caused by mutations in the FVIII gene. We have used Southern blotting and chemical mismatch analysis (CMA) to identify the mutations causing haemophilia A in 59 local or referred patients or carriers of haemophilia A. Southern blot analysis of 87 families with FVIII : C < 5% identified 31 as positive for the intron 22 inversion. Analysis of 19 of the inversion-negative families and a further nine families with mild or moderate haemophilia A by CMA resulted in the identification of a heterogeneous spectrum of mutations in the FVIII gene comprising 21 single base-pair substitutions and nine deletions. Seventeen of the base-pair substitutions are missense, two nonsense, and two are splice-site mutations. Two patients were found to have compound mutations with two mutations identified on a single X chromosome. Six of the point mutations and six of the deletions have not been reported previously in the haemophilia A mutation database. Unusually, a missense mutation, as well as deletion and splice-site mutations, was found to be associated with exon-skipping events.

A new candidate missense mutation (Leu 1657 IIe) in an apparently asymptomatic type 2A (phenotype IIA) von Willebrand disease family.

Type 2A von Willebrand disease (VWD) is mostly an autosomal dominantly inherited bleeding disorder characterised by a qualitative defect of von Willebrand factor (VWF). Mutation screening was used to screen the whole of VWF gene followed by direct sequencing to detect the mutation in a father and son diagnosed with type 2A (phenotype IIA) von Willebrand disease. A C5219 to A transversion was detected predicting Leucine to Isoleucine substitution in codon 1657. This novel missense mutation which was also identified by MboI restriction enzyme analysis, was found in both patient and his father but not in any other unaffected family member or 50 unrelated normal individuals. This substitution was reproduced by in vitro site directed mutagenesis of full-length VWF cDNA and transiently expressed in COS-7 cells. The corresponding recombinant VWF protein exhibited the full spectrum of VWF multimers, suggesting that the abnormal multimer seen in the patient results from increased proteolysis.

Gender and the stigma of onchocercal skin disease in Africa.

This paper reports results from a multicenter study of gender differences in the stigma associated with onchocercal skin disease (OSD) in five African sites: Cameroon, Ghana, Nigeria (Awka and Ibadan) and Uganda. The studies used a common protocol to compare affected and unaffected respondents, that is, men and women with onchodermatitis in highly endemic areas and respondents from communities with low endemicity or no onchocerciasis. The methods were both quantitative and qualitative, allowing for the comparison of stigma scores and people's verbal descriptions of their experiences and attitudes. Questions to the unaffected were asked after providing them with photographs and short descriptions (vignettes) depicting typical cases. We found that stigma was expressed more openly by the unaffected, who perceived OSD as something foreign or removed from themselves, whereas the affected tended to deny that they experienced stigma as a result of the condition. Gender differences in stigma scores were not significantly different for men and women, but qualitative data revealed that stigma was experienced differently by men and women, and that men and women were affected by it in distinctive ways. Men were more concerned about the impact of the disease on sexual performance and economic prospects, whereas women expressed more concern about physical appearance and life chances, especially marriage. Similar trends were found in the different sites in the responses of affected and unaffected respondents, and differences between them, despite geographical and cultural variations.

Independent occurrence of the novel Arg2163 to His mutation in the factor VIII gene in three unrelated families with haemophila A with different phenotypes. Mutations in brief no. 126. Online.

Using chemical mismatch analysis or denaturing gradient gel electrophoresis followed by nucleotide sequencing, we have identified the same G6545A mutation leading to an Arg2163 His subsitution in the factor VIII gene of three haemophiliacs from unrelated families. One of the affected individuals has severe haemophilia, while the other two are moderately severe. While we cannot exclude the possibility that these differences in phenotype arise from differences in VIII:C assay methods, other studies have also identified different clinical phenotypes in individuals with the same mutations, and suggested that they may arise from extragenic factors that affect or modify gene expression or protein function. The G6545A mutation occurs at a CG dinucleotide which is a known mutation hotspot, and which may explain the independent occurrence in unrelated families.

De novo mutation causing sporadic type 2A von Willebrancd's disease: report of three cases.

Chemical mismatch detection has been used to screen selectively part of the A2 domain of exon 28 of the von Willebrand factor gene of three unrelated patients with apparently sporadic type 2A von Willebrand disease (vWD) and their parents and siblings. Mismatches have been defined by DNA sequencing and mutations authenticated by restriction enzyme analysis. While a mutation was identified in all three patients, no evidence of mutation could be found in their asymptomatic/un-affected parents or siblings, proving the disease to be truly sporadic in these cases. Of these, two with severe clinical bleeding had a serine 743 to leucine substitution while the third patient with clinically less severe bleeding had an arginine 834 to tryptophan substitution. The possible genetic mechanisms for sporadic type 2A vWD in these families are discussed.

Fundamentals of the polymerase chain reaction: a future in clinical diagnostics?

The polymerase chain reaction (PCR) is an example of a technique that is having a profound impact on both fundamental and applied clinical science research. The availability of PCR-based diagnostic kits for the detection of polymorphisms within the HLA-DQA1 locus portends a technology that will undoubtedly become part of the clinical laboratory's diagnostic arsenal, and will extend and/or refine laboratory-based diagnosis in many areas. With current research effort directed to increase our knowledge of the overall structure of the human genome, and the identification of disease-associated genes and sequences, we can anticipate correspondingly rapid advances in its applications. This paper briefly reviews the basic facets of the PCR, which suggest it will fulfill such a role in future clinical diagnosis.