Diversity of Culicoides in the middle belt of Ghana with Implications on the transmission of Mansonella perstans; a molecular approach.

BACKGROUND: Culicoides, also known as biting midges, carry pathogens which include Mansonella perstans. Mansonella perstans is a nematode parasite implicated in a number of disease outcomes. Even though a high prevalence of about 75% M. perstans infection has been recorded in some communities in the middle belt of Ghana, and a wide diversity of Culicoides species has been identified, the exact Culicoides species transmitting M. perstans in Ghana has not yet been deciphered. This study therefore aimed at assessing the species diversity of Culicoides and their role in the transmission of M. perstans in the middle belt of Ghana. METHODS: Culicoides species were sampled from 11 communities in the Asante-Akim North and Sene West districts in the middle belt of Ghana. Centre for Disease Control (CDC) UV light traps, as well as human bait (i.e. human landing catch and engorged catch) methods were used to assess the species abundance and diversity of Culicoides in the study communities in the wet and dry season. A colorimetric Loop-Mediated Isothermal Amplification (LAMP) assay was performed to assess the vector competence of the various Culicoides species. RESULTS: A total of 4810 Culicoides from 6 species were sampled. These included Culicoides inornatipennis, C. milnei, C. schultzei, C. grahamii, C. neavei, and C. imicola. Culicoides imicola was the most abundant species (56%) followed by C. grahamii (16%). Light traps sampled the most diverse species (6 species). Human landing catch and engorged catch methods identified three anthropophilic species, C. grahamii, C. milnei, and C. inornatipennis, with C. grahamii being the most anthropophilic with a peak biting time between the hours of 5 p.m. to 6 p.m. Generally, there was relatively higher species abundance in the wet than dry season. LAMP assay identified C. grahamii as the potential vector for M. perstans transmission in the middle belt of Ghana. CONCLUSIONS: For the first time, we have demonstrated that C. grahamii is the potential competent vector for M. perstans transmission in the middle belt of Ghana. It is more abundant in the rainy season and has a peak biting time between the hours of 5 and 6 p.m.

Differences in PPD- and mitogen-induced T-cell activation marker expression characterize immunopathology in acute tuberculosis patients.

Impaired T-cell responses to mitogens and high T-cell activation marker (TAM) expression on Mycobacterium tuberculosis-specific T-cells characterize immunopathology in patients with tuberculosis (TB). In a study of patients with TB (n = 60) and asymptomatic contacts (controls, n = 37), we found that TB patients had higher CD38+ T-cell proportions specific for M. tuberculosis protein (PPDMtb), yet total proportions of PPDMtb-specific T-cells were comparable. Notably, both activated (CD38+) and total IFN-γ+ T-cells from TB patients had lower mitogen (phytohemagglutinin, PHA)-induced responses. This impaired mitogen response improved the classification efficacy of the TAM-TB assay, especially employing the PPD/PHA-induced T-cell ratio.

The Role of Maternal Depression Symptoms and Maternal Attachment in Predicting Exclusive Breastfeeding: A Multisite Prospective Study.

Background: Previous research shows that 61% of children younger than 6 months in low- and middle-income countries (LMICs) are not exclusively breastfed. Although data on the role of pre- and postnatal depression on breastfeeding exclusivity is mixed, fetomaternal attachment might foster breastfeeding exclusivity. Thus, we tested the potential mediating role of fetomaternal attachment and postnatal depression in the relationship between maternal prenatal depression and exclusive breastfeeding. Materials and Methods: Data were collected as part of a prospective, cross-cultural project, Evidence for Better Lives Study, which enrolled 1208 expectant mothers, in their third trimester of pregnancy across eight sites, from LMICs. Of the whole sample, 1185 women (mean age = 28.32, standard deviation [SD] = 5.77) completed Computer-Aided Personal Interviews on prenatal depressive symptoms, fetomaternal attachment, and socioeconomic status. A total of 1054 women provided follow-up data at 3-6 months after birth, about postnatal depressive symptoms, exclusive breastfeeding, and infant health indicators. Path analysis was used to assess parallel mediation. Results: In the whole sample, the effect of prenatal depression on breastfeeding exclusivity was completely mediated by postnatal depression, whereas fetomaternal attachment did not mediate the relationship. The full mediation effect was replicated individually in Pakistan and Sri Lanka. Conclusions: The study results indicate that prenatal depression symptoms contributed to the development of depressive symptoms after birth, negatively affecting the probability of exclusive breastfeeding. Future research should explore this in early prevention interventions, increasing the chances of healthy child development in LMICs. Considering the mixed results around the sites, it is important to better understand the relationship between maternal depression, fetomaternal attachment and breastfeeding behavior in each site's socio-cultural context.

Diminished Interleukin-7 receptor expression on T-cell subsets in tuberculosis patients.

Immunopathology in human tuberculosis affects T-cell phenotype and functions. Previous studies identified impaired T-cell sensitivity to Interleukin (IL)-7 accompanied by lower IL-7 receptor α-chain (IL-7Rα) expression in patients with acute tuberculosis. In the present study, we characterized affected T-cell subsets and determined the influence of tuberculosis disease severity and treatment response. Tuberculosis patients (n = 89) as well as age- and gender-matched asymptomatic contacts (controls, n = 47) were recruited in Ghana. Mycobacterium (M.) tuberculosis sputum burden was monitored prior to and during treatment. Blood samples from all patients and controls were analyzed for IL-7Rα expression and T-cell markers by multi-colour flow cytometry. CD4+ and CD8+ T-cells of tuberculosis patients showed generally lower IL-7Rα expression as compared to controls. Concomitantly, tuberculosis patients had higher proportions of naïve and lower proportions of memory CD4+ T-cells. Notably, a subset of CD27 positive central memory T-cells (Tcm), which lacked IL-7Rα expression was enriched in tuberculosis patients as compared to controls. M. tuberculosis sputum burden was not associated with differences in IL-7Rα expression. Treatment duration and response showed no clear effects although IL-7Rα expression patterns were highly variable. These results suggested generally impaired generation of memory CD4+ T-cells and enrichment of a Tcm subset without IL-7Rα expression in patients with tuberculosis.

Monocyte pathology in human tuberculosis is due to plasma milieu changes and aberrant STAT signalling.

Monocyte-derived macrophages contribute centrally to immune protection in Mycobacterium tuberculosis infection and changes in monocyte phenotype characterize immunopathology in tuberculosis patients. Recent studies highlighted an important role of the plasma milieu in tuberculosis immunopathology. Here, we investigated monocyte pathology in patients with acute tuberculosis and determined tuberculosis plasma milieu effects on phenotype as well as cytokine signalling of reference monocytes. Patients with tuberculosis (n = 37) and asymptomatic contacts (controls n = 35) were recruited as part of a hospital-based study in the Ashanti region of Ghana. Multiplex flow cytometry phenotyping of monocyte immunopathology was performed and effects of individual blood plasma samples on reference monocytes prior to and during treatment were characterized. Concomitantly, cell signalling pathways were analysed to elucidate underlying mechanisms of plasma effects on monocytes. Multiplex flow cytometry visualization characterized changes in monocyte subpopulations and detected higher expression of CD40, CD64 and PD-L1 in monocytes from tuberculosis patients as compared to controls. Aberrant expression normalized during anti-mycobacterial treatment and also CD33 expression decreased markedly. Notably, higher CD33, CD40 and CD64 expression was induced in reference monocytes when cultured in the presence of plasma samples from tuberculosis patients as compared to controls. STAT signalling pathways were affected by the aberrant plasma milieu and higher levels of STAT3 and STAT5 phosphorylation was found in tuberculosis plasma-treated reference monocytes. Importantly, high pSTAT3 levels were associated with high CD33 expression and pSTAT5 correlated with CD40 as well as CD64 expression. These results suggested plasma milieu effects with potential implications on monocyte phenotype and function in acute tuberculosis.

BCG-Vaccinated Children with Contact to Tuberculosis Patients Show Delayed Conversion of Mycobacterium tuberculosis-Specific IFN-γ Release.

Mycobacterium (M.) bovis BCG vaccination is recommended for healthy babies after birth in several countries with a high prevalence of tuberculosis, including Ghana. Previous studies showed that BCG vaccination prevents individuals from developing severe clinical manifestations of tuberculosis, but BCG vaccination effects on the induction of IFN-γ after M. tuberculosis infection have hardly been investigated. Here, we performed IFN-γ-based T-cell assays (i.e., IFN-γ Release Assay, IGRA; T-cell activation and maturation marker assay, TAM-TB) in children who had contact with index tuberculosis patients (contacts). These contacts were classified as either being BCG vaccinated at birth (n = 77) or non-BCG-vaccinated (n = 17) and were followed up at three timepoints for a period of one year to determine immune conversion after M. tuberculosis exposure and potential infection. At baseline and month 3, BCG-vaccinated contacts had significantly lower IFN-γ levels after stimulation with M. tuberculosis-specific proteins as compared to non-BCG-vaccinated contacts. This resulted in decreased proportions of positive IGRA results (BCG-vaccinated: 60% at baseline, 57% at month 3; non-BCG-vaccinated: 77% and 88%, respectively) at month 3. However, until month 12, immune conversion in BCG-vaccinated contacts led to balanced proportions in IGRA responders and IFN-γ expression between the study groups. TAM-TB assay analyses confirmed higher proportions of IFN-γ-positive T-cells in non-BCG-vaccinated contacts. Low proportions of CD38-positive M. tuberculosis-specific T-cells were only detected in non-BCG-vaccinated contacts at baseline. These results suggest that BCG vaccination causes delayed immune conversion as well as differences in the phenotype of M. tuberculosis-specific T-cells in BCG-vaccinated contacts of tuberculosis patients. These differences are immune biomarker candidates for protection against the development of severe clinical tuberculosis manifestations.

Impaired T-cell response to phytohemagglutinin (PHA) in tuberculosis patients is associated with high IL-6 plasma levels and normalizes early during anti-mycobacterial treatment.

PURPOSE: Human tuberculosis is characterized by immunopathology that affects T-cell phenotype and functions. Previous studies found impaired T-cell response to phytohemagglutinin (PHA) in patients with acute tuberculosis. However, the influence of disease severity, affected T-cell subsets, and underlying mechanisms remain elusive. METHODS: Here we investigated PHA-induced and antigen-specific T-cell effector cytokines in tuberculosis patients (n = 55) as well as in healthy asymptomatic contacts (n = 32) from Ghana. Effects of Mycobacterium (M.) tuberculosis sputum burden and treatment response were analyzed and compared during follow-up. Finally, cytokine characteristics of the aberrant plasma milieu in tuberculosis were analyzed as a potential cause for impaired PHA response. RESULTS: PHA-induced IFN-γ expression was significantly lower in sputum-positive tuberculosis patients as compared to both, contacts and paucibacillary cases, and efficiently discriminated the study groups. T-cell responses to PHA increased significantly early during treatment and this was more pronounced in tuberculosis patients with rapid treatment response. Analysis of alternative cytokines revealed distinct patterns and IL-22, as well as IL-10, showed comparable expression to IFN-γ in response to PHA. Finally, we found that high IL-6 plasma levels were strongly associated with impaired IFN-γ and IL-22 response to PHA. CONCLUSION: We conclude that impaired T-cell response to PHA stimulation in acute tuberculosis patients (i) was potentially caused by the aberrant plasma milieu, (ii) affected differentially polarized T-cell subsets, (iii) normalized early during treatment. This study shed light on the mechanisms of impaired T-cell functions in tuberculosis and yielded promising biomarker candidates for diagnosis and monitoring of treatment response.

Plasma cytokine levels characterize disease pathogenesis and treatment response in tuberculosis patients.

BACKGROUND: Mycobacterium (M.) tuberculosis-caused immunopathology is characterized by aberrant expression of plasma cytokines in human tuberculosis. Disease severity and long-term anti-mycobacterial treatment are potentially influenced by immunopathology and normalization of plasma cytokine levels during therapy may indicate treatment efficacy and recovery. STUDY DESIGN AND METHODS: In this study, we analyzed the concentrations of selected plasma cytokines (i.e., IL-6, IP-10, IL-10, IL-22, IFNγ, GM-CSF, IL-8) and M. tuberculosis sputum burden in patients with tuberculosis (n = 76). Cytokine levels were compared to healthy contacts (n = 40) and changes under treatment were monitored (i.e., 6 and 16 weeks after treatment start). According to differences in M. tuberculosis sputum burden and conversion, tuberculosis patients were classified as paucibacillary as well as 'rapid' or 'slow' treatment responders. A subgroup of tuberculosis patients had fatal disease courses. RESULTS: Six of seven cytokines were significantly higher in tuberculosis patients as compared to contacts and four of these (i.e., IL-6, IP-10, IL-10, and IL-22) were detectable in the majority of tuberculosis patients. IL-6 showed the strongest discriminating capacity for tuberculosis disease and in combination with IL-10 concentrations efficiently classified paucibacillary tuberculosis cases as well as those with fatal disease outcome. In addition, IL-6 and IP-10 levels decreased significantly after 6 weeks of treatment and analyses of subgroups with differential treatment response showed delayed decline of IL-6 levels in slow treatment responders. CONCLUSIONS: Combinations of different plasma cytokine (namely, IL-6, IL-10, and IP-10) efficiently classified tuberculosis patients with differential mycobacterial burden and especially IL-6 qualified as a biomarker candidate for early treatment response.

Physiological, anthropometric parameters, and balance skill response of healthy bankers to fitness training.

Sedentary lifestyle as a predisposing factor of chronic diseases like hypertension, diabetes, stroke and obesity is a common phenomenon in the banking job. Studies suggest that fitness training improves health of bankers but has not been established among Ghanaian bankers. This study examined the physiological, anthropometric parameters, and balance skill responses of relatively healthy bankers to fitness training. Twelve bankers aged 28 to 55 years (36.41±7.16 years) in Kumasi completed a 6-month fitness training program (FTP) of 30-min gym workouts and 1-hr swimming per session. Physiological, anthropometric parameters, and balance skill variables assessments were conducted in three trials: pretraining, midtraining and post-FTP. FTP caused significant decrease in pre-post systolic blood pressure (P=0.001), diastolic blood pressure (P=0.000), heart rate (P=0.006), waist circumference (P= 0.007), waist-to-hip ratio (P=0.007), and bone density (P=0.038). There was significance decrease in body mass index (P=0.047) between pre- and midtraining status. Weight significantly decreased among the three trials (P=0.017). Pre-post opened (P=0.043) and closed (P=0.015) eye balance skills increased significantly. Effects of FTP were significantly higher in female (P<0.05). Participants who were at the stage 1 and 2 hypertensions pretraining became normotensive posttraining. Six months FTP has beneficial effects on the physiological, anthropometric parameters, and balance skill of relatively healthy bankers.

Perceived satisfaction with health services under National Health Insurance Scheme: Clients' perspectives.

The study aims to explore the perceived satisfaction of insured clients in financing health services through National Health Insurance in Ghana. A quantitative method was used to recruit 380 respondents, selected by multistage cluster sampling. Data were collected through the administration of questionnaires. More than half, 57.9%, of respondents were males, and the average age was 34 years. Most respondents, 74.3%, were insured. Overall, 53.12% of insured clients were dissatisfied with the services of providers. Factors, such as benefit package of insurance, willingness to pay higher premium, and perceived discrimination were significantly associated with poor satisfaction with health services. The current advocacy for and awareness about the use of health insurance as a prepayment plan should be prioritised in policy initiatives. The benefit package for the insurance should be increased in order to cover all disease conditions that afflict the Ghanaian population.

Effectiveness of combined and conventional exercise trainings on the biochemical responses of stroke patients.

Stroke is the topmost cause of mortality and disability in Ghana. Conventional exercise is mostly used aside pharmacological management technique but the complimentary effects of combined exercise training (CET) have not been reported. This study investigated the effectiveness of combined and conventional exercise trainings on lipid-cardiovascular profiles and body composition among stroke patients in Kumasi. Thirteen stroke survivors between ages 35-68 years (mean±standard deviation: age, 59.88±10.88 years; duration of illness, 18.11±7.72 years) participated in the study. Participants were randomly assigned to CET and conventional exercise group. The CET had 10 weeks of 3 types of exercise trainings, 3 days/wk; conventional exercise group monitored for daily conventional activities during the same period. Post weight (P= 0.003), body mass index (P=0.004), systolic blood pressure (P=0.022), diastolic blood pressure (P=0.004), heart rate (P=0.003), and total cholesterol (P=0.044) of the CET were significantly improved. CET significantly improved total cholesterol (P=0.005) and low-density lipoprotein (P=0.006) better than the conventional exercises. Effectiveness of CET to positively enhance biochemical responses in the management of strove patients was established.

Total lymphocyte count as a surrogate marker for CD4 count in resource-limited settings.

BACKGROUND: CD4 testing is the recognized gold standard used to stage HIV/AIDS, guide treatment decisions for HIV-infected persons and evaluate effectiveness of therapy. The need for a less expensive surrogate marker that can be used in resource-limited setting is however necessary. The study sought to assess the suitability of Total lymphocyte count (TLC) as a surrogate marker for CD4 count in resource-limited localities in Ghana. METHODS: This observational study was conducted at the Central Regional Hospital, which has one of the established antiretroviral therapy centres in Ghana. A total of one hundred and eighty-four (184) confirmed HIV I seropositive subjects were included in the study. Blood samples were taken from all the subjects for estimation of CD4 and total lymphocyte counts. The study subjects were further categorised into three (3) groups according to the Centers for Disease Control and Prevention (CDC) classification criteria as follows: CD4 counts (1) ≥ 500 cells/mm3 (2) 200-499 cells/mm3 and (3) <200 cells/mm3. Positive predictive value (PPV), negative predictive value (NPV), sensitivity and specificity of various TLC cut-offs were computed for three groups. Correlation and Receiver Operator Characteristic analysis was performed for the various CD4 counts and their corresponding Total Lymphocyte count obtained. RESULTS: The sensitivity, specificity, positive and negative predictive values of TLC 1200 cells/ mm3 to predict CD4 count were <200 cells/mm3 72.2%, 100%, 100% and 95.7% respectively. A TLC of 1500 cells/ mm3 was found to have maximal sensitivity (96.67%), specificity (100%), PPV (100%) and NPV (75.0%) for predicting a CD4 cell count of 200-499 cell/mm3. A TLC of 1900 cells/mm3 was also found to have a maximal sensitivity (98.45%), specificity (100%), PPV (100%) and NPV (100%) for predicting CD4 count ≥500 cells/mm3. A positive correlation was noted between 184 paired CD4 and TLC counts (r = 0.5728). CONCLUSION: Total Lymphocyte count can therefore adequately serve as a surrogate marker for CD4 count in HIV patients who are naïve for antiretroviral therapy in resource-limited areas.