Psychological Distress and Its Associated Factors Among Parents of Children With Congenital Heart Disease: A Cross-Sectional Mixed Method Study at the National Cardiothoracic Center, Ghana.

BACKGROUND: The aim of this study was to examine the prevalence and factors associated with psychological distress among parents of children with congenital heart disease (PCCHD) receiving treatment at the National Cardiothoracic Centre (NCTC), Accra. METHODS: A cross-sectional mixed method study involving 248 parents of children with congenital heart disese receiving treatment at the NCTC was employed. Systematic sampling was used to select participants. The outcome variables were depression, anxiety, and somatization, assessed using the brief symptom inventory. A semistructured interview guide was used to understand the contextual factors that contribute to parents' psychological distress. Poisson regression and inductive thematic analyses were employed for quantitative and qualitative data, respectively. RESULTS: The prevalence of depression, anxiety, and somatization was 42.9% [95%CI = 36.7-49.3], 42.4% [95%CI = 36.3-48.9], and 41.2% [95%CI = 35.1-47.6] respectively. Parents who were divorced [aPR (95%CI) = 1.60(1.02-2.51)], hadonly basic level education [aPR (95%CI) = 2.13(1.34-3.39)], were without a religious affiliation [aPR (95%CI)= 2.24(1.58-3.19)], and in the low asset quintile significantly increased the likelihood of psychological distress. Delay/postponement of scheduled surgery, worsening state of child's condition, feeling a sense of helplessness, societal stigma, and lack of funding or health insurance for the treatment of the condition were contextual factors that contributed to increased psychological distress among parents. CONCLUSION: The prevalence of psychological distress was quite high among parents of children with congenital heart disease in Ghana and was associated with both parent and patient factors. Psychosocial care should be integrated into the clinical health services of this group of patients and their parents to improve psychological health outcomes.

Satisfaction levels and associated influencing factors among inpatients and outpatients with breast cancer at a tertiary health facility in Ghana.

Objectives: To uncover variables linked to breast cancer patient satisfaction in order to improve policy choices and actions for breast cancer care in Ghana. Design: We employed a cross-sectional design using a quantitative approach. Setting: The Radiotherapy, Oncology and Surgery Departments of the Korle Bu Teaching Hospital, Accra. Participants: Inpatient and outpatient breast cancer patients. Main outcome measures: The level of inpatient and outpatient satisfaction was measured using descriptive and inferential statistical analyses. The Shapiro-Wilk test was employed to assess normality, while the Heckman selection model assessed significance with outcomes of interest. Results: A total of 636 participants, with a mean age of 52.64±14.07 years, were recruited. The measured inpatient and outpatient levels of satisfaction out of 100 were 74.06±7.41 and 49.99±1.00 respectively, while the self-reported satisfaction levels out of 5 were 4.22±0.63 and 4.11±0.85 respectively. The level of inpatient satisfaction was significantly influenced by age, marital status, income level, and number of previous facilities visited (p<0.05). Outpatient satisfaction level was significantly associated with place of residence and income level (p<0.05). Conclusions: The study offers insight into the satisfaction levels of breast cancer patients receiving inpatient and outpatient services at the largest tertiary referral centre and teaching hospital in Ghana, as well as the factors influencing attendance and satisfaction levels. Understanding and improving breast cancer patients' levels of satisfaction is a way that providers can safeguard their emotional well-being. Improvement in patient satisfaction at our institution among outpatients is an area for future growth. Funding: Gardner-Holt Women's Health Grant program, Centre for Global Surgery 2021.

Self-reported smoking, urine cotinine, and risk of type 2 diabetes: Findings from the PREVEND prospective cohort study.

BACKGROUND: Smoking is a major risk factor for type 2 diabetes (T2D), but the evidence has mostly relied on self-reports. We aimed to compare the associations of smoking exposure as assessed by self-reports and urine cotinine with T2D. METHODS: Using the PREVEND prospective study, smoking status was assessed at baseline by self-reports and urine cotinine in 4708 participants (mean age, 53 years) without a history of diabetes. Participants were classified as never, former, light current and heavy current smokers according to self-reports and analogous cut-offs for urine cotinine. Hazard ratios (HRs) with 95% CIs were estimated for T2D. RESULTS: During a median follow-up of 7.3 years, 259 participants developed T2D. Compared with self-reported never smokers, the multivariable adjusted HRs (95% CI) of T2D for former, light current, and heavy current smokers were 1.02 (0.75-1.4), 1.41 (0.89-2.22), and 1.30 (0.88-1.93), respectively. The corresponding adjusted HRs (95% CI) were 0.84 (0.43-1.67), 1.61 (1.12-2.31), and 1.58 (1.08-2.32), respectively, as assessed by urine cotinine. Urine cotinine-assessed but not self-reported smoking status improved T2D risk prediction beyond established risk factors. CONCLUSION: Urine cotinine assessed smoking status may be a stronger risk indicator and predictor of T2D compared to self-reported smoking status.

Macrophage susceptibility to infection by Ghanaian Mycobacterium tuberculosis complex lineages 4 and 5 varies with self-reported ethnicity.

Background: The epidemiology of Mycobacterium tuberculosis complex (MTBC) lineage 5 (L5) infections in Ghana revealed a significantly increased prevalence in Ewes compared to other self-reported ethnic groups. In that context, we sought to investigate the early phase of tuberculosis (TB) infection using ex vivo infection of macrophages derived from the blood of Ewe and Akan ethnic group volunteers with MTBC L4 and L5 strains. Methods: The study participants consisted of 16 controls, among which self-reported Akan and Ewe ethnicity was equally represented, as well as 20 cured TB cases consisting of 11 Akans and 9 Ewes. Peripheral blood mononuclear cells were isolated from both healthy controls and cured TB cases. CD14+ monocytes were isolated and differentiated into monocyte-derived macrophages (MDMs) before infection with L4 or L5 endemic strains. The bacterial load was assessed after 2 hours (uptake) as well as 3 and 7 days post-infection. Results: We observed a higher capacity of MDMs from Ewes to phagocytose L4 strains (p < 0.001), translating into a higher bacillary load on day 7 (p < 0.001) compared to L5, despite the higher replication rate of L5 in Ewe MDMs (fold change: 1.4 vs. 1.2, p = 0.03) among the controls. On the contrary, within macrophages from Akans, we observed a significantly higher phagocytic uptake of L5 (p < 0.001) compared to L4, also translating into a higher load on day 7 (p = 0.04). However, the replication rate of L4 in Akan MDMs was higher than that of L5 (fold change: L4 = 1.2, L4 = 1.1, p = 0.04). Although there was no significant difference in the uptake of L4 and L5 among cured TB cases, there was a higher bacterial load of both L4 (p = 0.02) and L5 (p = 0.02) on day 7 in Ewe MDMs. Conclusion: Our results suggest that host ethnicity (driven by host genetic diversity), MTBC genetic diversity, and individual TB infection history are all acting together to modulate the outcome of macrophage infections by MTBC.

A three-year review of lung cancer patient characteristics in a tertiary hospital.

Objective: The study sought to determine clinical characteristics and histologic subtypes of a cohort of lung cancer patients in a tertiary facility. Design: Retrospective review of the medical records of histology-confirmed lung cancer cases at the respiratory clinic over a 3-year period. Setting: Respiratory Clinic, Korle-Bu Teaching Hospital, Accra, Ghana. Participants: All adult patients with histologically diagnosed lung cancer were enrolled. Main outcome measures: Lung cancer histological types. Results: The proportion of lung cancer cases was 12.4%. The majority were women (57.8%) and the mean age at diagnosis was 55.8±16.0 years. The patients were predominantly non-smokers (61%). Common symptoms were chronic cough and chest pain. More than two-thirds of the cases presented in clinical stages III and IV with the predominant histological subtype being adenocarcinoma in smokers and non-smokers. Genetic testing for epidermal growth factor receptor (EGFR) and Anaplastic Lymphoma kinase (ALK) mutations were largely absent. Conclusions: The majority of lung cancer patients presented late with advanced disease. Adenocarcinoma was the predominant histological subtype in a predominantly non-smoking population, with an increased prevalence among women less than 60 years. This should encourage testing for genetic mutations to improve patient survival. Funding: None declared.

Marijuana use and suicidal behaviours among school-going adolescents in Africa: assessments of prevalence and risk factors from the Global School-Based Student Health Survey.

BACKGROUND: Marijuana use among adolescents, including high school students, has been consistently reported to be associated with a high incidence of suicidal behaviours. Little empirical research has been conducted on the propensity impact of marijuana use on suicidal behaviours in Africa. AIMS: To assess factors associated with marijuana use and further quantify marijuana use as an associated factor of suicidal behaviours, including repeated attempted suicide, suicidal ideation and suicide planning, among high school students in Africa. METHODS: A cross-sectional study was conducted among 32 802 school-going adolescents using the Global School-Based Student Health Survey data from 10 African countries grouped into West Africa, North Africa, South-East Africa, South Africa and East Africa subregions. Marijuana use and repeated attempted suicide were the main outcome variables. We employed double selection least absolute shrinkage and selection operator poisson regression model to assess risk factors associated with marijuana use and dominance analysis to establish ranked important and common risk factors. Inverse probability weighting poisson regression adjustment was applied to assess impact. RESULTS: The prevalence of marijuana use and repeated attempted suicide were 3.7% (95% CI: 3.1 to 4.3) and 6.6% (95% CI: 5.9 to 7.4), respectively. The most important risk factor for marijuana use generally across the countries and specifically in three subregions was alcohol consumption, which constituted approximately 40% of the impact. The average treatment effect on the treated (ATT) indicated that marijuana use significantly increased the risk of suicidal ideation, suicide planning and repeated attempted suicide by 12% (ATT=0.12 (95% CI: 0.02 to 0.22)), 18% (ATT=0.18 (95% CI: 0.13 to 0.22)) and 31% (ATT=0.31 (95% CI: 0.20 to 0.41)), respectively. CONCLUSIONS: Marijuana use was significantly associated with suicidal behaviours (suicidal ideation, planning and repeated attempted suicide) among the students. To achieve Sustainable Development Goal 3.5 (to strengthen prevention and treatment of substance abuse), school-based psychosocial interventions should be streamlined to adequately assess and manage marijuana use. Targeting the most dominant risk factors in this population could translate into the reduction of suicidal behaviours in countries within Africa.

Unmet Needs of Healthcare Services and Associated Factors among a Cohort of Ghanaian Adults: A Nationally Stratified Cross-Sectional Study Design.

We estimated the prevalence of unmet needs of healthcare services (UNHS) and its associated factors among a cohort of older Ghanaian adults. World Health Organization (WHO) Study on Global AGEing and Adult Health for Ghana was used with a total of 4735 participants. Logistics regression analysis was performed using Stata 16 to assess associated factors. The overall UNHS was 3.7% (95% CI = 2.7-4.8) and the prevalence was significantly high amongst older adults aged 60 to 69 years (5.9%). Could not afford the healthcare (56.4%) was the main contender for UNHS. UNHS was influenced by; those aged 60 to 69 years [OR (95% CI) = 1.86 (1.19-2.91)]; no formal educational [aOR (95% CI) = 4.71 (1.27-17.38)], and no NHIS [OR (95% CI) = 1.78 (1.03-3.09)]. Participants needed care for joint pain (25.4%), and communicable diseases (19.1%). The inability to access healthcare was relatively higher for older adults more advanced in age, with low education, and for those without health insurance. Health system strengthening including financial protection by expanding the National Health Insurance Scheme to all Ghanaians in line with Ghana's Universal Health Coverage Roadmap would reduce the unmet healthcare needs of older adults.

Utilization of eye health services and diabetic retinopathy: a cross-sectional study among persons living with diabetes visiting a tertiary eye care facility in Ghana.

BACKGROUND: There have been a major advance made in screening, early diagnosis, and prompt treatment of Diabetic Retinopathy among Person living with diabetes (PLWD). However, screening services remain a challenge in Low-Middle-Income-Countries where access to eye care professionals is inadequate. This study assesses the utilization of Eye Health Service prevalence (UEHS) among PLWD and associated factors and further quantifies its association with Non-Proliferative Diabetic Retinopathy (NPDR). METHODS: A cross-sectional study design with a random sample of 360 PLWD was conducted at Korle-Bu Teaching Hospital, a National Referral Centre in Ghana from May to July 2019. UEHS and DR were the study outcomes. We adopted Poisson and Probit regression analysis to assess factors associated with UEHS over the past year. We employed pairwise and phi correlation (fourfold correlational analysis) to assess the relationship between UEHS and DR (ordinal and binary respectively). Ordered Logistic and Poisson regression were applied to assess the association between the UEHS and DR. Stata 16.1 was used to perform the analyses and a p-value ≤ 0.05 was deemed significant. RESULTS: The prevalence of UEHS over the past year and DR was 21.7 %(95 %CI = 17.7-26.2) and 65.0 %(95 %CI = 59.9-69.8 respectively. The prevalence of severe NPDR with Clinically Significant Macular Edema (CSME) was 23.9 %(19.8-28.6). Type of diabetes, increasing age, educational level, mode of payment for healthcare services, marital status, years since diagnosis, and current blood glucose significantly influenced UEHS. There was a negative relationship between DR and UEHS (Pairwise and φ correlation were - 20 and - 15 respectively; p < 0.001). Non-UEHS among PLWD doubles the likelihood of experiencing severe NPDR with CSME compared with UEHS among PLWD [aOR(95 %CI) = 2.05(1.03-4.08)]. Meanwhile, the prevalence of DR among patients per non-UEHS was insignificantly higher [12 %; aPR(95 %CI) = 0.89-1.41)] compared with patients who utilized eye care health service. CONCLUSIONS: Most of the PLWD did not utilize the eye health service even once in a year and that was highly influenced by type of diabetes and increasing age. Type 2 diabetes patients and middle age decreased the likelihood of UEHS. There was a negative relationship between DR and UEHS among PLWD and this doubled the likelihood of experiencing severe NPDR with CSME. Structured health education and screening interventions are key to improving UEHS.

Flexible bronchoscopy in a tertiary healthcare facility: a review of indications and outcomes.

Objectives: Flexible Fibreoptic bronchoscopy (FFB) is a major diagnostic and therapeutic tool employed largely in respiratory medicine but its use in our country has been quite limited. We performed a retrospective review of the indications, overall diagnostic yield and safety of FFB at the Korle-Bu Teaching Hospital (KBTH). Study Design: Retrospective study. Study Setting: Cardiothoracic Unit, Korle-Bu Teaching Hospital. Study Participants: All bronchoscopy records from January 2017 - December 2018. Interventions: Eight-five bronchoscopy reports generated over a 2-year period were reviewed. Using a data extraction form, patient's demographic details, indications for FFB, sedation given, specimen obtained and results of investigation, and complications encountered were recorded and entered into SPSS version 22. Descriptive analysis was performed and presented as means and percentages. Results: Suspected lung cancer was the predominant indication for bronchoscopy requests (55.3%). Diagnostic yield of endobronchial biopsy was 86.7% increased to 93.3% when biopsy was combined with bronchial washing cytology. Bronchial washing geneXpert was positive in 20.8% of sputum negative cases, and 20.7% of patients with unresolved pneumonia and bronchiectasis had a positive microbial yield. Overall mild complications occurred in 5.9% of patients with no mortality. Conclusion: Flexible bronchoscopy has a significantly high diagnostic yield, particularly in evaluating lung cancers and undiagnosed lung infections with minimal associated complications, hence increasing its availability in the country and widening the diagnostic scope at the cardiothoracic unit of the Korle-Bu Teaching Hospital. Funding: None declared.

Marijuana use and repeated attempted suicide among senior high school students in Ghana: Evidence from the WHO Global School-Based Student Health Survey, 2012.

BACKGROUND: The association between substance use including marijuana use and attempted suicide has been well documented. However, little is known about marijuana use and its association with attempted suicide repetition among young people in low-income and middle-income contexts. AIMS: This analysis was conducted to assess the factors associated with marijuana use and ascertain marijuana use as a determinant of repeated attempted suicide among senior high school (SHS) students in Ghana. METHODS: Data from the 2012 Global School-Based Student Health Survey in Ghana was used for this study. Modified Poisson, Logistic and Probit models weighted with Mahalanobis distance matching within propensity calliper were employed separately to determine the hypothetical association between marijuana use and repeated attempted suicide. All analysis was performed using Stata 16 and p≤0.05 was deemed statistically significant. RESULTS: The prevalence estimates of marijuana use and repeated attempted suicide among SHS students in Ghana were 3.4% (95% CI: 2.3 to 5.1) and 11.5% (95% CI: 9.1 to 14.4), respectively. The prevalence of marijuana use was significantly associated with school grade, smoking exposure, parent smoker, alcohol intake and truancy. Marijuana use was positively associated with repeated attempted suicide among SHS in Ghana (φ correlation=0.23, p<0.001). Repeated attempted suicide among students who use marijuana was approximately threefold and fivefold significant compared with non-marijuana use students, based on the Poisson (adjusted prevalence ratio: 3.02; 95% CI: 1.67 to 5.43, p<0.001) and Logistic (adjusted OR:5.06; 95% CI: 3.19 to 11.64, p<0.001) estimates respectively. Also, the Probit model showed that marijuana use significantly increased the log count of repeated attempted suicide by 95% (aß: 0.95; 95% CI: 0.49 to 1.41, p<0.001). CONCLUSION: Marijuana use does not only influence the onset of suicidal attempts but also repeated attempted suicide among SHS students in Ghana. Special attention is required for suicide attempters with a history of repeated attempts and current marijuana use among SHS students in Ghana. Early identification of the potential risk and protective factors is recommended to inform school-based interventions. National level structured school-based substance abuse interventions and health promotion programmes would be useful.

Leishmanicidal Potential of Hardwickiic Acid Isolated From Croton sylvaticus.

Leishmania is a parasitic protozoon responsible for the neglected tropical disease Leishmaniasis. Approximately, 350 million people are susceptible and close to 70,000 death cases globally are reported annually. The lack of effective leishmanicides, the emergence of drug resistance and toxicity concerns necessitate the pursuit for effective antileishmanial drugs. Natural compounds serve as reservoirs for discovering new drugs due to their chemical diversity. Hardwickiic acid (HA) isolated from the stembark of Croton sylvaticus was evaluated for its leishmanicidal potential against Leishmania donovani and L. major promastigotes. The susceptibility of the promastigotes to HA was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide/phenazine methosulfate colorimetric assay with Amphotericin B serving as positive control. HA showed a significant antileishmanial activity on L. donovani promastigotes with an IC50 value of 31.57± 0.06 µM with respect to the control drug, amphotericin B with IC50 of 3.35 ± 0.14 µM). The cytotoxic activity was observed to be CC50 = 247.83 ± 6.32 µM against 29.99 ± 2.82 µM for curcumin, the control, resulting in a selectivity index of SI = 7.85. Molecular modeling, docking and dynamics simulations of selected drug targets corroborated the observed antileishmanial activity of HA. Novel insights into the mechanisms of binding were obtained for trypanothione reductase (TR), pteridine reductase 1 (PTR1), and glutamate cysteine ligase (GCL). The binding affinity of HA to the drug targets LmGCL, LmPTR1, LdTR, LmTR, LdGCL, and LdPTR1 were obtained as -8.0, -7.8, -7.6, -7.5, -7.4 and -7.1 kcal/mol, respectively. The role of Lys16, Ser111, and Arg17 as critical residues required for binding to LdPTR1 was reinforced. HA was predicted as a Caspase-3 stimulant and Caspase-8 stimulant, implying a possible role in apoptosis, which was shown experimentally that HA induced parasite death by loss of membrane integrity. HA was also predicted as antileishmanial molecule corroborating the experimental activity. Therefore, HA is a promising antileishmanial molecule worthy of further development as a biotherapeutic agent.

Alcohol consumption and fruits and vegetable intake among older adults in Ghana: a cross-sectional survey based on WHO-SAGE Wave 2 data.

BACKGROUND: Alcohol consumption and inadequate fruits and vegetable (FnV) intake are major reasons for the shift from communicable to non-communicable diseases (NCDs) over the years. The older Ghanaian adult is at high risk of NCD and data on alcohol and FnV consumption are required to guide policy to mitigate its effect. This analysis aimed to determine the factors associated with alcohol consumption and assess the relationship between alcohol consumption and FnV intake among Ghanaians aged 50 years and older. METHODS: This analysis used WHO Study on Global Ageing and Adult Health (SAGE) Wave 2, Ghana data set conducted between 2014 and 2015. Data on demographic characteristics, FnV intake, and alcohol consumption were collated and analysed. Multivariable Poisson, logistic and probit regression analyses were performed to assess the associations between alcohol consumption and inadequate FnV intake. RESULTS: A total of 3533 Ghanaians aged 50 years and older, 41.0% men and 59.0% women, were included in this study. The prevalence of lifetime alcohol consumption was 22.8% (95% CI 20.7% to 25.1%). Alcohol consumption was significantly associated with sex, age group, marital status, religion, place of residence and history of smoking. The prevalence of adequate FnV intake was 52.6% with a mean daily intake of 6.45 servings: 2.98 for fruits and 3.47 for vegetables. There was a significant positive correlation between inadequate FnV intake and alcohol consumption. Inadequate FnV consumption was significantly higher among lifetime alcohol consumers compared with non-alcohol consumers. (Poisson estimate; adjusted Prevalence Ratio (aPR) (95% CI)=1.35 (1.12 to 1.63), logistic estimate; adjusted Old Ratio (aOR) (95% CI)=1.13 (1.05 to 1.21) and probit estimate; adjusted normalized coefficient (aß) (95% CI)=0.19 (0.07 to 0.31)). CONCLUSION: About a quarter and nearly half of older Ghanaian adults consume alcohol and inadequate FnV, respectively. Alcohol consumption is significantly associated with inadequate FnV intake. Interventions to address inadequate FnV intake among older adults in Ghana should also include policies that regulate the use of alcohol in this population.

High Levels of IL-10 and CD4+CD25hi+ Treg Cells in Endemic Burkitt's Lymphoma Patients.

BACKGROUND: The interplay between Epstein-Barr virus infection, malaria, and endemic Burkitt's Lymphoma is not well understood. Reports show diminished EBV-specific Th1 responses in children living in malaria endemic areas and deficiency of EBNA1-specific IFN-γ T cell responses in children with endemic Burkitt's Lymphoma (eBL). This study, therefore, examined some factors involved in the loss of EBNA-1-specific T cell responses in eBL. METHODS: T-cell subset frequencies, activation, and IFN-γ- or IL-4-specific responses were analyzed by flow-cytometry. Plasma cytokine levels were measured by ELISA. RESULTS: CD4+ and CD8+ cells in age- and sex-matched healthy controls (n = 3) expressed more IFN-γ in response to all immunostimulants than in pediatric endemic BL (eBL) patients (n = 4). In healthy controls, IFN-γ expression was higher than IL-4 expression, whereas in eBL patients the expression of IL-4 by CD4+ cells to EBNA-1 was slightly higher than IFN-γ. Moreover, the blood levels of TNF-α was significantly lower (p = 0.004) while IL-10 was significantly higher (p = 0.038), in eBL patients (n = 21) compared to controls (n = 16). Additionally, the frequency of CD4+CD25hi+ T cells was higher in both age-matched acute uncomplicated malaria (n = 26) and eBL (n = 14) patients compared to healthy controls (n = 19; p = 0.000 and p = 0.027, respectively). CONCLUSION: The data suggest that reduced Th1 response in eBL might be due to increased levels of IL-10 and T reg cells.

Cytokine response to selected MTB antigens in Ghanaian TB patients, before and at 2 weeks of anti-TB therapy is characterized by high expression of IFN-γ and Granzyme B and inter- individual variation.

BACKGROUND: There has been a long held belief that patients with drug-susceptible TB are non-infectious after two weeks of therapy. Recent microbiological and epidemiological evidence has challenged this dogma, however, the nature of the Mtb-specific cellular immune response during this period has not been adequately investigated. This knowledge could be exploited in the development of immunological biomarkers of early treatment response. METHODS: Cellular response to four Mtb infection phase-dependent antigens, ESAT-6/CFP-10 fusion protein and three DosR encoded proteins (Rv1733c, Rv2029c, Rv2628) were evaluated in a Ghanaian TB cohort (n=20) before and after 2 weeks of anti TB therapy. After 6-days in vitro stimulation, Peripheral blood mononuclear cell (PBMC) culture supernatant was harvested and the concentration of IFN-γ, Granzyme B, IL-10, IL-17, sIL2Rα and TNF-α were determined in a 6-plex Luminex assay. Frequencies of IFN-γ + CD4 and CD8 T cells were also determined in an intracellular cytokine assay. RESULTS: All antigens induced higher levels of IFN-γ, followed by Granzyme B, TNF-α and IL-17 and low levels of IL-10 and sIL-2R-α in PBMC before treatment and after 2 weeks of treatment. Median cytokine levels of IFN-γ, Granzyme B, IL-17 and sIL-2R-α increased during week two, but it was significant for only Rv1733-specific production of Granzyme B (P = 0. 013). The median frequency of antigen specific IFN-γ + CD4 T cells increased at week two; however, only the increase in the ESAT-6/CFP-10-specific response was significant (P = 0. 0008). In contrast, the median frequency of ESAT-6/CFP-10- specific IFN-γ + CD8 T cell responses declined during week two (P = 0. 0024). Additionally, wide inter-individual variation with three distinct patterns were observed; increase in all cytokine levels, decrease in all cytokine levels and fluctuating cytokine levels after 2 weeks of treatment. CONCLUSION: The second week of effective chemotherapy was characterized by a general increase in cytokine response to Mtb-specific antigens suggestive of an improvement in cellular response with therapy. However, the wide inter-individual variation observed would limit the utility of cytokine biomarkers during this period.

An integrated buccal delivery system combining chitosan films impregnated with peptide loaded PEG-b-PLA nanoparticles.

Peptide (insulin) loaded nanoparticles (NPs) have been embedded into buccal chitosan films (Ch-films-NPs). These films were produced by solvent casting and involved incorporating in chitosan gel (1.25% w/v), NPs-Insulin suspensions at three different concentrations (1, 3, and 5mg of NPs per film) using glycerol as plasticiser. Film swelling and mucoadhesion were investigated using 0.01M PBS at 37°C and texture analyzer, respectively. Formulations containing 3mg of NPs per film produced optimised films with excellent mucoadhesion and swelling properties. Dynamic laser scattering measurements showed that the erosion of the chitosan backbone controlled the release of NPs from the films, preceding in vitro drug (insulin) release from Ch-films-NPs after 6h. Modulated release was observed with 70% of encapsulated insulin released after 360h. The use of chitosan films yielded a 1.8-fold enhancement of ex vivo insulin permeation via EpiOral™ buccal tissue construct relative to the pure drug. Flux and apparent permeation coefficient of 0.1µg/cm(2)/h and 4×10(-2)cm(2)/h were respectively obtained for insulin released from Ch-films-NPs-3. Circular dichroism and FTIR spectroscopy demonstrated that the conformational structure of the model peptide drug (insulin) released from Ch-films-NPs was preserved during the formulation process.

Insights into deregulated TNF and IL-10 production in malaria: implications for understanding severe malarial anaemia.

BACKGROUND: Severe malarial anaemia (SMA) is a major life-threatening complication of paediatric malaria. Protracted production of pro-inflammatory cytokines promoting erythrophagocytosis and depressing erythropoiesis is thought to play an important role in SMA, which is characterized by a high TNF/IL-10 ratio. Whether this TNF/IL-10 imbalance results from an intrinsic incapacity of SMA patients to produce IL-10 or from an IL-10 unresponsiveness to infection is unknown. Monocytes and T cells are recognized as the main sources of TNF and IL-10 in vivo, but little is known about the activation status of those cells in SMA patients. METHODS: The IL-10 and TNF production capacity and the activation phenotype of monocytes and T cells were compared in samples collected from 332 Ghanaian children with non-overlapping SMA (n = 108), cerebral malaria (CM) (n = 144) or uncomplicated malaria (UM) (n = 80) syndromes. Activation status of monocytes and T cells was ascertained by measuring HLA-DR+ and/or CD69+ surface expression by flow cytometry. The TNF and IL-10 production was assessed in a whole-blood assay after or not stimulation with lipopolysaccharide (LPS) or phytohaemaglutinin (PHA) used as surrogate of unspecific monocyte and T cell stimulant. The number of circulating pigmented monocytes was also determined. RESULTS: Monocytes and T cells from SMA and CM patients showed similar activation profiles with a comparable decreased HLA-DR expression on monocytes and increased frequency of CD69+ and HLA-DR+ T cells. In contrast, the acute-phase IL-10 production was markedly decreased in SMA compared to CM (P = .003) and UM (P = .004). Although in SMA the IL-10 response to LPS-stimulation was larger in amplitude than in CM (P = .0082), the absolute levels of IL-10 reached were lower (P = .013). Both the amplitude and levels of TNF produced in response to LPS-stimulation were larger in SMA than CM (P = .019). In response to PHA-stimulation, absolute levels of IL-10 produced in SMA were lower than in CM (P = .005) contrasting with TNF levels, which were higher (P = .001). CONCLUSIONS: These data reveal that SMA patients have the potential to mount efficient IL-10 responses and that the TNF/IL-10 imbalance may reflect a specific monocyte and T cell programming/polarization pattern in response to infection.

Cerebral malaria is associated with low levels of circulating endothelial progenitor cells in African children.

Damage to the cerebral microvasculature is a feature of cerebral malaria. Circulating endothelial progenitor cells are needed for microvascular repair. Based on this knowledge, we hypothesized that the failure to mobilize sufficient circulating endothelial progenitor cells to the cerebral microvasculature is a pathophysiologic feature of cerebral malaria. To test this hypothesis, we compared peripheral blood levels of CD34 (+)/VEGFR2(+) and CD34 (+)/CD133(+) cells and plasma levels of the chemokine stromal cell-derived growth factor 1 (SDF-1) in 214 children in Accra, Ghana. Children with cerebral malaria had lower levels of CD34 (+)/VEGFR2(+) and CD34 (+)/CD133(+) cells compared with those with uncomplicated malaria, asymptomatic parasitemia, or healthy controls. SDF-1 levels were higher in children with acute malaria compared with healthy controls. Together, these results uncover a potentially novel role for endothelial progenitor cell mobilization in the pathophysiology of cerebral malaria.

Selective activation of TCR-gammadelta+ cells in endemic Burkitt's lymphoma.

BACKGROUND: The overlap in geographical distribution of Plasmodium falciparum malaria and endemic Burkitt's lymphoma (eBL)--an aggressive Epstein-Barr virus (EBV)-associated B-cell tumour occurring almost exclusively in the tropics--strongly suggests a link between the two diseases. It is suspected that the polyclonal B-cell activation in P. falciparum malaria may precipitate a breakdown in homeostatic T-cell control of EBV-immortalized B-cell proliferation. Previous studies have suggested that a particular T-cell subset, characterized by expression of Vdelta1+ gammadelta T-cell receptors, is important for maintaining B-cell homeostasis, both in P. falciparum- exposed populations and in individuals subject to polyclonal B-cell activation of other aetiology. The objective of the present study was, therefore, to characterize lymphocyte phenotypes and to investigate possible differences in T-cell subset composition and activation status in P. falciparum-exposed Ghanaian children with and without eBL. METHODS: Venous blood samples in heparin from 21 eBL patients (mean age: 7.0 years; range: 3-11 years), referred to the Burkitt's Tumour Centre at Korle-Bu Teaching Hospital, Accra and 15 healthy, age and sex matched children, were stained with fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)-, R-phycoerythrin (RPE)- and RPE-Cy5-conjugated antibodies (CD3, CD4, CD8, CD25, CD69, CD95, HLA-DR, TCR-gammadelta, Vdelta1, Vdelta3, Vgamma9 and B-cells) and acquired on a flow cytometer. RESULTS: A reduction in the proportion of CD3+ cells in eBL patients, due mainly to perturbations among TCR-gammadelta+ cells was observed. In contrast, the proportions of CD4+ or CD8+ cells were relatively unaffected, as were the mean numbers of peripheral blood mononuclear cells. CONCLUSION: Selective changes in numbers and activation status of TCR-gammadelta+ cells occurs in Ghanaian children with eBL, a pattern which is similar to P. falciparum-induced changes. The data supports the hypothesis of a regulatory role for Vdelta1+ TcR-gammadelta T-cells in maintaining B-cell homeostasis and provides insights into the pathogenesis of eBL.

Increased levels of inflammatory mediators in children with severe Plasmodium falciparum malaria with respiratory distress.

BACKGROUND: Respiratory distress (RD), a symptom of underlying metabolic acidosis, has been identified as a major risk factor for mortality in children with severe malaria in Africa, yet the molecular mediators involved in the pathogenesis of RD have not been identified. METHODS: We studied circulating levels of mediators of inflammation--including the cytokines tumor necrosis factor (TNF)- alpha and interleukin (IL)-10; the chemokines macrophage inflammatory protein (MIP)-1 alpha , MIP-1 beta , and IL-8; and the immune activation marker neopterin--in children with RD, severe malarial anemia (SMA), cerebral malaria (CM), and uncomplicated malaria (UM). RESULTS: Children with RD had significantly higher plasma levels of TNF- alpha , IL-10, and neopterin and a significantly higher TNF- alpha : IL-10 ratio than those without RD. In addition, the results demonstrated that, relative to UM, CM was associated with increased levels of TNF- alpha and decreased levels of MIP-1 alpha , whereas SMA was associated with decreased levels of IL-10. Circulating levels of neopterin were inversely correlated with hemoglobin, whereas levels of MIP-1 beta were positively correlated with parasitemia. CONCLUSIONS: We conclude that distinct clinical presentations of severe malaria are associated with specific patterns of inflammatory mediators. In particular, we show, to our knowledge for the first time, that patients with malaria and RD have a strong and unbalanced proinflammatory response that may be involved in the pathogenesis of the underlying metabolic acidosis.

Characteristics of hepatitis B virus in Ghana: full length genome sequences indicate the endemicity of genotype E in West Africa.

Hepatitis B virus (HBV) genotypes from A to H have distinct geographical distributions and have been shown to affect the clinical features as well as the course of the HBV infection. HBV genotype E has been found only in Africa. However, the complete genomes of this genotype, which were isolated mainly from West Africa, were available only for a few samples. In this study, five full-length genomes and seven other small S genes of HBV strains from Ghanaian blood donors were sequenced and investigated. Following phylogenetic analysis, all of the Ghanaian HBV strains were clustered closely in genotype E. All of the 12 small S genes showed the same characteristic of subtype ayw4. The complete genomes of the five Ghanaian strains showed marked similarity with each other and with the reported genotype E strains (96.7%-99.1%). Genotype E strains showed low intra-genotypic diversity (1.8%) and carried the conserved signature pattern in pre-S1 as well as in the full genome sequence. Of note, the finding of the G145R escape mutant in an unvaccinated Ghanaian blood donor might raise concern as to the ongoing nation-wide hepatitis B vaccination program in Ghana.