Genetic diversity and microevolution in clinical Cryptococcus isolates from Cameroon.

Cryptococcal meningitis is the second most common cause of death in people living with HIV/AIDS, yet we have a limited understanding of how cryptococcal isolates change over the course of infection. Cryptococcal infections are environmentally acquired, and the genetic diversity of these infecting isolates can also be geographically linked. Here, we employ whole genome sequences for 372 clinical Cryptococcus isolates from 341 patients with HIV-associated cryptococcal meningitis obtained via a large clinical trial, across both Malawi and Cameroon, to enable population genetic comparisons of isolates between countries. We see that isolates from Cameroon are highly clonal, when compared to those from Malawi, with differential rates of disruptive variants in genes with roles in DNA binding and energy use. For a subset of patients (22) from Cameroon, we leverage longitudinal sampling, with samples taken at days 7 and 14 post-enrollment, to interrogate the genetic changes that arise over the course of infection, and the genetic diversity of isolates within patients. We see disruptive variants arising over the course of infection in several genes, including the phagocytosis-regulating transcription factor GAT204. In addition, in 13% of patients sampled longitudinally, we see evidence for mixed infections. This approach identifies geographically linked genetic variation, signatures of microevolution, and evidence for mixed infections across a clinical cohort of patients affected by cryptococcal meningitis in Central Africa.

Cryptococcal meningitis, caused by Cryptococcus, results in approximately half a million deaths per year globally. We compare clinical Cryptococcus samples from Cameroon and Malawi to explore the genetic diversity of these isolates. We find instances of mixed-strain infections and identify genetic variants arising in Cryptococcus over disease.

Cryptococcal Antigen in Serum and Cerebrospinal Fluid for Detecting Cryptococcal Meningitis in Adults Living With Human Immunodeficiency Virus: Systematic Review and Meta-Analysis of Diagnostic Test Accuracy Studies.

Cryptococcal antigen (CrAg) detection could direct the timely initiation of antifungal therapy. We searched MEDLINE and Embase for studies where CrAg detection in serum/cerebrospinal fluid (CSF) and CSF fungal culture were done on adults living with human immunodeficiency virus (HIV) who had suspected cryptococcal meningitis (CM). With Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2), we evaluated the risk of bias in 11 included studies with 3600 participants, and used a random-effects meta-analysis to obtain summary sensitivity and specificity of serum and CSF CrAg, as well as agreement between CSF CrAg and CSF culture. Summary sensitivity and specificity of serum CrAg were 99.7% (97.4-100) and 94.1% (88.3-98.1), respectively, and summary sensitivity and specificity of CSF CrAg were 98.8% (96.2-99.6) and 99.3% (96.7-99.9), respectively. Agreement between CSF CrAg and CSF culture was 98% (97-99). In adults living with HIV who have CM symptoms, serum CrAg negativity may rule out CM, while positivity should prompt induction antifungal therapy if lumbar puncture is not feasible. In a first episode of CM, CSF CrAg positivity is diagnostic.

Infertility and associated factors in three hospitals in Douala, Cameroon: a cross-sectional study.

AIM: We determined the prevalence and factors associated with couple infertility in three hospitals in Douala, Cameroon. METHODS: We conducted a cross-sectional study from December 18th 2015 to March 18th 2016 in three public hospitals in Douala. Three hundred and sixty participants were studied prospectively for associated factors using a multivariate logistic regression model and 4732 files were studied retrospectively for the prevalence of infertility. Statistical significance was set at p < 0.05. RESULTS: The prevalence of couple infertility was 19.2%. In logistic models, the factors which independently increased the risk of couple infertility were a history of reproductive tract infection/STI, a history of uterine fibroids, a history of dysmenorrhea and abortion for the females while for males it was a history of mumps, erectile dysfunction and exposure to chemicals/toxic substances/pesticides. CONCLUSION: One in every five couples in this study was infertile. Several factors affect the risks associated with couple infertility. The identification of these factors could help detect subgroups of couples at high risk of infertility. Reproductive health education, screening programmes for STI's that may lead to infertility should be offered to couples.

Addition of Flucytosine to Fluconazole for the Treatment of Cryptococcal Meningitis in Africa: A Multicountry Cost-effectiveness Analysis.

BACKGROUND: Mortality from cryptococcal meningitis remains very high in Africa. In the Advancing Cryptococcal Meningitis Treatment for Africa (ACTA) trial, 2 weeks of fluconazole (FLU) plus flucytosine (5FC) was as effective and less costly than 2 weeks of amphotericin-based regimens. However, many African settings treat with FLU monotherapy, and the cost-effectiveness of adding 5FC to FLU is uncertain. METHODS: The effectiveness and costs of FLU+5FC were taken from ACTA, which included a costing analysis at the Zambian site. The effectiveness of FLU was derived from cohorts of consecutively enrolled patients, managed in respects other than drug therapy, as were participants in ACTA. FLU costs were derived from costs of FLU+5FC in ACTA, by subtracting 5FC drug and monitoring costs. The cost-effectiveness of FLU+5FC vs FLU alone was measured as the incremental cost-effectiveness ratio (ICER). A probabilistic sensitivity analysis assessed uncertainties and a bivariate deterministic sensitivity analysis examined the impact of varying mortality and 5FC drug costs on the ICER. RESULTS: The mean costs per patient were US $847 (95% confidence interval [CI] $776-927) for FLU+5FC, and US $628 (95% CI $557-709) for FLU. The 10-week mortality rate was 35.1% (95% CI 28.9-41.7%) with FLU+5FC and 53.8% (95% CI 43.1-64.1%) with FLU. At the current 5FC price of US $1.30 per 500 mg tablet, the ICER of 5FC+FLU versus FLU alone was US $65 (95% CI $28-208) per life-year saved. Reducing the 5FC cost to between US $0.80 and US $0.40 per 500 mg resulted in an ICER between US $44 and US $28 per life-year saved. CONCLUSIONS: The addition of 5FC to FLU is cost-effective for cryptococcal meningitis treatment in Africa and, if made available widely, could substantially reduce mortality rates among human immunodeficiency virus-infected persons in Africa.

New Insights Into Cryptococcus Spp. Biology and Cryptococcal Meningitis.

PURPOSE OF REVIEW: Defective cell-mediated immunity is a major risk factor for cryptococcosis, a fatal disease if untreated. Cryptococcal meningitis (CM), the main presentation of disseminated disease, occurs through hematogenous spread to the brain from primary pulmonary foci, facilitated by yeast virulence factors. We revisit remarkable recent improvements in the prevention, diagnosis and management of CM. RECENT FINDINGS: Cryptococcal antigen (CrAg), main capsular polysaccharide of Cryptococcus spp. is detectable in blood and cerebrospinal fluid of infected patients with point of care lateral flow assays. Recent World Health Organization guidelines recommend 7-day amphotericin B plus flucytosine, then 7-day high dose (1200 mg/day) fluconazole for induction treatment of HIV-associated CM. Management of raised intracranial pressure, a consequence of CM, should rely mainly on daily therapeutic lumbar punctures until normalisation. In HIV-associated CM, following introduction of antifungal therapy, (re)initiation of antiretroviral therapy should be delayed by 4-6 weeks to prevent immune reconstitution inflammatory syndrome, common in CM. CM is a fatal disease whose diagnosis has recently been simplified. Treatment should always include antifungal combination therapy and management of raised intracranial pressure. Screening for immune deficiency should be mandatory in all patients with cryptococcosis.

Case fatality rate and viral aetiologies of acute respiratory tract infections in HIV positive and negative people in Africa: The VARIAFRICA-HIV systematic review and meta-analysis.

BACKGROUND: To set priorities for efficient control of acute respiratory tract infection (ARTI) in Africa, it is necessary to have accurate estimate of its burden, especially among HIV-infected populations. OBJECTIVES: To compare case fatality rate (CFR) and viral aetiologies of ARTI between HIV-positive and HIV-negative populations in Africa. STUDY DESIGN: We searched PubMed, EMBASE, Web of Knowledge, Africa Journal Online, and Global Index Medicus to identify studies published from January 2000 to April 2018. Random-effect meta-analysis method was used to assess association (pooled weighted odds ratios (OR) with 95% confidence interval (CI)). RESULTS: A total of 36 studies (126,526 participants) were included. CFR was significantly higher in patients with HIV than in HIV-negative controls (OR 4.10, 95%CI: 2.63-6.27, I²: 93.7%). The risk was significantly higher among children ≤5 years (OR 5.51, 95%CI 2.83-10.74) compared to people aged >5 years (OR 1.48, 95%CI 1.17-1.89); p = 0.0002. There was no difference between children (15 years) and adults and between regions of Africa. There was no difference for viral respiratory aetiologies (Enterovirus, Adenovirus, Bocavirus, Coronavirus, Metapneumovirus, Parainfluenza, Influenza, and Respiratory Syncytial Virus) of ARTI between HIV-positive and HIV-negative people, except for Rhinovirus where being HIV-negative was associated with Rhinovirus (OR 0.70; 95%CI 0.51-0.97, I²: 63.4%). CONCLUSIONS: This study shows an increased risk of deaths among HIV-infected individuals with ARTI, however with no difference in viral aetiologies compared to HIV-negative individuals in Africa. ARTI deserves more attention from HIV health-care providers for efficient control. Specific strategies are needed for HIV-positive children under 5.

Recent advances in managing HIV-associated cryptococcal meningitis.

The recent development of highly sensitive and specific point-of-care tests has made it possible to diagnose HIV-associated cryptococcal meningitis within minutes. However, diagnostic advances have not been matched by new antifungal drugs and treatment still relies on old off-patent drugs: amphotericin B, flucytosine and fluconazole. Cryptococcal meningitis treatment is divided in three phases: induction, consolidation and maintenance. The induction phase, aimed at drastically reducing cerebrospinal fluid fungal burden, is key for patient survival. The major challenge in cryptococcal meningitis management has been the optimisation of induction phase treatment using the limited number of available medications, and major progress has recently been made. In this review, we summarise data from key trials which form the basis of current treatment recommendations for HIV-associated cryptococcal meningitis.

Healthcare Costs and Life-years Gained From Treatments Within the Advancing Cryptococcal Meningitis Treatment for Africa (ACTA) Trial on Cryptococcal Meningitis: A Comparison of Antifungal Induction Strategies in Sub-Saharan Africa.

BACKGROUND: Mortality from cryptoccocal meningitis remains high. The ACTA trial demonstrated that, compared with 2 weeks of amphotericin B (AmB) plus flucystosine (5FC), 1 week of AmB and 5FC was associated with lower mortality and 2 weeks of oral flucanozole (FLU) plus 5FC was non-inferior. Here, we assess the cost-effectiveness of these different treatment courses. METHODS: Participants were randomized in a ratio of 2:1:1:1:1 to 2 weeks of oral 5FC and FLU, 1 week of AmB and FLU, 1 week of AmB and 5FC, 2 weeks of AmB and FLU, or 2 weeks of AmB and 5FC in Malawi, Zambia, Cameroon, and Tanzania. Data on individual resource use and health outcomes were collected. Cost-effectiveness was measured as incremental costs per life-year saved, and non-parametric bootstrapping was done. RESULTS: Total costs per patient were US $1442 for 2 weeks of oral FLU and 5FC, $1763 for 1 week of AmB and FLU, $1861 for 1 week of AmB and 5FC, $2125 for 2 weeks of AmB and FLU, and $2285 for 2 weeks of AmB and 5FC. Compared to 2 weeks of AmB and 5FC, 1 week of AmB and 5FC was less costly and more effective and 2 weeks of oral FLU and 5FC was less costly and as effective. The incremental cost-effectiveness ratio for 1 week of AmB and 5FC versus oral FLU and 5FC was US $208 (95% confidence interval $91-1210) per life-year saved. CLINICAL TRIALS REGISTRATION: ISRCTN45035509. CONCLUSIONS: Both 1 week of AmB and 5FC and 2 weeks of Oral FLU and 5FC are cost-effective treatments.

Impact of Routine Cryptococcal Antigen Screening and Targeted Preemptive Fluconazole Therapy in Antiretroviral-naive Human Immunodeficiency Virus-infected Adults With CD4 Cell Counts <100/µL: A Systematic Review and Meta-analysis.

Cryptococcal antigen (CrAg) screening and targeted preemptive fluconazole in antiretroviral-naive human immunodeficiency virus-infected adults with CD4 cell counts <100/µL seems promising as a strategy to reduce the burden of cryptococcal meningitis (CM). We searched MEDLINE, EMBASE, and Web of Science and used random-effect meta-analysis to assess the prevalence of blood CrAg positivity (31 studies; 35644 participants) and asymptomatic CM in CrAg-positive participants and the incidence of CM and the all-cause mortality rate in screened participants. The pooled prevalence of blood CrAg-positivity was 6% (95% confidence interval [CI], 5%-7%), and the prevalence of asymptomatic CM in CrAg-positive participants was 33% (95% CI, 21%-45%). The incidence of CM was 21.4% (95% CI, 11.6%-34.4%) without preemptive fluconazole and 5.7% (95% CI, 3.0%-9.7%) with preemptive fluconazole therapy initiated at 800 mg/d. In CrAg-positive participants, postscreening lumbar puncture before initiating preemptive fluconazole at 800 mg/d further reduced the incidence of CM to null and showed some survival benefits. However, the all-cause mortality rate remained significantly higher in CrAg-positive than in CrAg-negative participants (risk ratio, 2.2; 95% CI, 1.7-2.9; P < .001).

Leave no one behind: response to new evidence and guidelines for the management of cryptococcal meningitis in low-income and middle-income countries.

In 2018, WHO issued guidelines for the diagnosis, prevention, and management of HIV-related cryptococcal disease. Two strategies are recommended to reduce the high mortality associated with HIV-related cryptococcal meningitis in low-income and middle-income countries (LMICs): optimised combination therapies for confirmed meningitis cases and cryptococcal antigen screening programmes for ambulatory people living with HIV who access care. WHO's preferred therapy for the treatment of HIV-related cryptococcal meningitis in LMICs is 1 week of amphotericin B plus flucytosine, and the alternative therapy is 2 weeks of fluconazole plus flucytosine. In the ACTA trial, 1-week (short course) amphotericin B plus flucytosine resulted in a 10-week mortality of 24% (95% CI -16 to 32) and 2 weeks of fluconazole and flucytosine resulted in a 10-week mortality of 35% (95% CI -29 to 41). However, with widely used fluconazole monotherapy, mortality because of HIV-related cryptococcal meningitis is approximately 70% in many African LMIC settings. Therefore, the potential to transform the management of HIV-related cryptococcal meningitis in resource-limited settings is substantial. Sustainable access to essential medicines, including flucytosine and amphotericin B, in LMICs is paramount and the focus of this Personal View.

Diagnostic and therapeutic strategies in cryptococcosis: impact on outcome.

Cryptococcosis diagnosis has been recently improved by the use of rapid cryptococcal antigen testing with lateral flow assays, which have proved sensitive and specific. Using "test and treat" screening strategies for cryptococcal disease with these tests has been showed effective in reducing cryptococcal meningitis (CM) in HIV-infected patients. Recommended induction, consolidation, and maintenance therapeutic strategy for CM is widely unavailable and/or expensive in low and middle-income settings. New therapeutic strategies, mostly using reduced duration, have recently shown acceptable outcome or are currently tested. Diagnostic and therapeutic guidelines for cryptococcal disease in limited resources countries are undergoing a paradigmatic shift.

Antifungal Combinations for Treatment of Cryptococcal Meningitis in Africa.

BACKGROUND: Cryptococcal meningitis accounts for more than 100,000 human immunodeficiency virus (HIV)-related deaths per year. We tested two treatment strategies that could be more sustainable in Africa than the standard of 2 weeks of amphotericin B plus flucytosine and more effective than the widely used fluconazole monotherapy. METHODS: We randomly assigned HIV-infected adults with cryptococcal meningitis to receive an oral regimen (fluconazole [1200 mg per day] plus flucytosine [100 mg per kilogram of body weight per day] for 2 weeks), 1 week of amphotericin B (1 mg per kilogram per day), or 2 weeks of amphotericin B (1 mg per kilogram per day). Each patient assigned to receive amphotericin B was also randomly assigned to receive fluconazole or flucytosine as a partner drug. After induction treatment, all the patients received fluconazole consolidation therapy and were followed to 10 weeks. RESULTS: A total of 721 patients underwent randomization. Mortality in the oral-regimen, 1-week amphotericin B, and 2-week amphotericin B groups was 18.2% (41 of 225), 21.9% (49 of 224), and 21.4% (49 of 229), respectively, at 2 weeks and was 35.1% (79 of 225), 36.2% (81 of 224), and 39.7% (91 of 229), respectively, at 10 weeks. The upper limit of the one-sided 97.5% confidence interval for the difference in 2-week mortality was 4.2 percentage points for the oral-regimen group versus the 2-week amphotericin B groups and 8.1 percentage points for the 1-week amphotericin B groups versus the 2-week amphotericin B groups, both of which were below the predefined 10-percentage-point noninferiority margin. As a partner drug with amphotericin B, flucytosine was superior to fluconazole (71 deaths [31.1%] vs. 101 deaths [45.0%]; hazard ratio for death at 10 weeks, 0.62; 95% confidence interval [CI], 0.45 to 0.84; P=0.002). One week of amphotericin B plus flucytosine was associated with the lowest 10-week mortality (24.2%; 95% CI, 16.2 to 32.1). Side effects, such as severe anemia, were more frequent with 2 weeks than with 1 week of amphotericin B or with the oral regimen. CONCLUSIONS: One week of amphotericin B plus flucytosine and 2 weeks of fluconazole plus flucytosine were effective as induction therapy for cryptococcal meningitis in resource-limited settings. (ACTA Current Controlled Trials number, ISRCTN45035509 .).

Long-term Mortality and Disability in Cryptococcal Meningitis: A Systematic Literature Review.

Cryptococcal meningitis (CM) is the primary cause of meningitis in adults with human immunodeficiency virus (HIV) infection and an emerging disease in HIV-seronegative individuals. No literature review has studied the long-term outcome of CM. We performed a systematic review on the long-term (≥3-month) impact of CM (Cryptococcus neoformans and Cryptococcus gattii) on mortality and disability in HIV-infected and non-HIV-infected adults. Although the quality of current evidence is limited, the long-term impact of CM on survival and disability seems to be high. One-year mortality ranged from 13% in an Australian non-HIV-infected C. gattii-infected cohort to 78% in a Malawian HIV-infected cohort treated with fluconazole monotherapy. One-year impairment proportions among survivors ranged from 19% in an Australian C. gattii cohort to >70% in a Taiwanese non-HIV- and HIV-infected cohorts. Ongoing early therapeutic interventions, early detection of impairments and access to rehabilitation services may significantly improve patients' survival and quality of life.

Diagnostic and therapeutic strategies in cryptococcosis: impact on outcome

Cryptococcosis diagnosis has been recently improved by the use of rapid cryptococcal antigen testing with lateral flow assays, which have proved sensitive and specific. Using "test and treat" screening strategies for cryptococcal disease with these tests has been showed effective in reducing cryptococcal meningitis (CM) in HIV-infected patients. Recommended induction, consolidation, and maintenance therapeutic strategy for CM is widely unavailable and/or expensive in low and middle-income settings. New therapeutic strategies, mostly using reduced duration, have recently shown acceptable outcome or are currently tested. Diagnostic and therapeutic guidelines for cryptococcal disease in limited resources countries are undergoing a paradigmatic shift.

Malnutrition in patients admitted to the medical wards of the Douala General Hospital: a cross-sectional study.

BACKGROUND: Malnutrition is common in acutely ill patients occurring in 30-50% of hospitalized patients. Awareness and screening for malnutrition is lacking in most health institutions in sub-Saharan Africa. This study aimed at screening for malnutrition using anthropometric and laboratory indices in patients admitted to the internal medicine wards. METHODS: A cross-sectional study. We screened for malnutrition in 251 consecutive patients admitted from January to March 2013 in the internal medicine wards. Malnutrition defined as body mass index (BMI) less than 18.5 kg/m2 and/or mid upper arm circumference (MUAC) less than 22 cm in women and 23 cm in men. Weight loss greater than 10% in the last 6 months prior to admission, relevant laboratory data, diagnosis at discharge and length of hospital stay (LOS) were also recorded. RESULTS: Mean age was 47 (SD 16) years. 52.6% were male. Mean BMI was 24.44 (SD 5.79) kg/m2 and MUAC was 27.8 (SD 5.0) cm. Median LOS was 7 (IQR 5-12) days. 42.4% of patients reported weight loss greater than 10% in the 6 months before hospitalization. MUAC and BMI correlated significantly (r = 0.78; p < 0.0001) and malnutrition by the two methods showed moderate agreement (κ = 0.56; p < 0.0001). Using the two methods in combination, the prevalence of malnutrition was 19.34% (35/251). Blood albumin and hemoglobin were significantly lower in malnourished patients. Malnourished patients had a significantly longer LOS (p = 0.019) when compared to those with no malnutrition. Malnutrition was most common amongst patients with malignancy. CONCLUSION: Malnutrition is common in patients admitted to the medical wards of the Douala General Hospital. Nutritional screening and assessment should be integrated in the care package of all admitted patients.

Characteristics of anti-hepatitis C virus antibody-positive patients in a hospital setting in Douala, Cameroon.

INTRODUCTION: Hepatitis C virus (HCV) infection is a major public health problem, especially in resource-limited settings where many patients are diagnosed at the stage of complications. In Cameroon, where HCV is endemic, little is known about the clinical, biological, and virological profile of HCV-infected patients. METHODS: A clinical case note review of all patients positive for antibodies against HCV diagnosed at the gastroenterology outpatient clinic of the Douala General Hospital, Cameroon, from January 2008 to December 2014, was performed. RESULTS: A total of 524 patients were included in the study, 53% of whom were female. The mean age was 56±13 years. A history of blood transfusion and a history of scarification were the most common potential risk factors for HCV exposure, as found in 16% and 13% of the study population, respectively. Current alcohol use was found in 24% of patients. Co-infection with hepatitis B virus and HIV was 3.6% and 3.4%, respectively. Among the patients, 39% had no complaint at diagnosis; only 16% were diagnosed through a routine medical checkup. Clinically, the most common finding was hepatomegaly (26.1% of patients). Transaminases above the upper limit of normal were found in 55.2% of patients, particularly those aged >57 years (p=0.001). Genotypes 1 (43.95%), 2 (25.11%), and 4 (28.25%) were the most common. Liver cirrhosis was present in 11% of patients and hepatocellular carcinoma in 4%, the latter being more common in males (p<0.001) and in those aged >57 years (p=0.03). CONCLUSIONS: In the gastroenterology clinic of Douala General Hospital, while almost 40% of patients who were anti-HCV antibody-positive were asymptomatic and diagnosed fortuitously, some already presented complications, including cirrhosis and hepatocellular carcinoma. There is an urgent need to put in place programs to increase awareness and diagnosis of HCV infection and to develop extensive and targeted anti-HCV treatment guidelines to improve the management of these patients in Cameroon.

The spectrum of cardiac disease in the West Region of Cameroon: a hospital-based cross-sectional study.

BACKGROUND: Cardiovascular disease is a growing public health problem in Africa. The extent of heart disease in Cameroon remains largely unknown. This study aimed at reporting the etiology of cardiac disease in a cardiologic clinic situated in a semi-urban area in the West region of Cameroon. METHODS: This is an analysis of echocardiographic diagnosis of cardiac disease done between July 2008 and October 2010 at the "Centre Medical de la Trinité" in the West region of Cameroon. Data included age, sex and echocardiographic findings. RESULTS: A total of 1252 patients presented with abnormal echocardiograms, 60.4% (n = 756) being female and 85.8% (n = 1074) aged over 50 years. Overall, the most important conditions were hypertensive heart disease (41.5%, n = 520) and cardiomyopathies (30.5%, n = 382). Among patients aged less than 10 years, congenital heart diseases were the most frequent (52.4%, n = 22), and rheumatic heart disease was the most important cardiac condition in patients aged 10 to 19 years (62.1%, n = 18) and those aged 20 to 39 years (53.3%, n = 8). Congenital heart diseases included persistent ductus arteriosus (27.6%, n = 8), tetralogy of Fallot (20.7%, n = 6) and inter-atria/interventricular communication (20.7%, n = 6). CONCLUSION: Hypertension is the leading cause of cardiac disease among the elderly in our setting, emphasizing the necessity to strengthen the preventive strategies against hypertension in Cameroon. Rheumatic heart disease and congenital heart disease frequent in children and youths highlight the need of early detection and treatment of throat infections, and of routine cardiac surgery services in Cameroon.