AP2A2 mutation and defective endocytosis in a Malian family with hereditary spastic paraplegia.

Hereditary spastic paraplegia (HSP) comprises a large group of neurogenetic disorders characterized by progressive lower extremity spasticity. Neurological evaluation and genetic testing were completed in a Malian family with early-onset HSP. Three children with unaffected consanguineous parents presented with symptoms consistent with childhood-onset complicated HSP. Neurological evaluation found lower limb weakness, spasticity, dysarthria, seizures, and intellectual disability. Brain MRI showed corpus callosum thinning with cortical and spinal cord atrophy, and an EEG detected slow background in the index patient. Whole exome sequencing identified a homozygous missense variant in the adaptor protein (AP) complex 2 alpha-2 subunit (AP2A2) gene. Western blot analysis showed reduced levels of AP2A2 in patient-iPSC derived neuronal cells. Endocytosis of transferrin receptor (TfR) was decreased in patient-derived neurons. In addition, we observed increased axon initial segment length in patient-derived neurons. Xenopus tropicalis tadpoles with ap2a2 knockout showed cerebral edema and progressive seizures. Immunoprecipitation of the mutant human AP-2-appendage alpha-C construct showed defective binding to accessory proteins. We report AP2A2 as a novel genetic entity associated with HSP and provide functional data in patient-derived neuron cells and a frog model. These findings expand our understanding of the mechanism of HSP and improve the genetic diagnosis of this condition.

Evaluation of Intussusception Following Pentavalent Rotavirus Vaccine (RotaTeq) Administration in 5 African Countries.

BACKGROUND: A low-level risk of intussusception following rotavirus vaccination has been observed in some settings and may vary by vaccine type. We examined the association between RotaTeq vaccination and intussusception in low-income settings in a pooled analysis from 5 African countries that introduced RotaTeq into their national immunization program. METHODS: Active surveillance was conducted at 20 hospitals to identify intussusception cases. A standard case report form was completed for each enrolled child, and vaccination status was determined by review of the child's vaccination card. The pseudo-likelihood adaptation of self-controlled case-series method was used to assess the association between RotaTeq administration and intussusception in the 1-7, 8-21, and 1-21 day periods after each vaccine dose in infants aged 28-245 days. RESULTS: Data from 318 infants with confirmed rotavirus vaccination status were analyzed. No clustering of cases occurred in any of the risk windows after any of the vaccine doses. Compared with the background risk of naturally occurring intussusception, no increased risk was observed after dose 1 in the 1-7 day (relative incidence = 2.71; 95% confidence interval [CI] = 0.47-8.03) or the 8-21 day window (relative incidence = 0.77; 95%CI = 0.0-2.69). Similarly, no increased risk of intussusception was observed in any risk window after dose 2 or 3. CONCLUSIONS: RotaTeq vaccination was not associated with increased risk of intussusception in this analysis from 5 African countries. This finding mirrors results from similar analyses with other rotavirus vaccines in low-income settings and highlights the need for vaccine-specific and setting-specific risk monitoring.

Endoscopic Third Ventriculostomy with Choroid Plexus Cauterization in Infantile Hydrocephalus: An Experience from Mali.

INTRODUCTION: Pediatric hydrocephalus is a common disease in sub-Saharan Africa. In Mali, 350-400 new cases are diagnosed in our center yearly. With a total land mass of 1,241,000 km2, patients in remote areas must travel up to 1,500 km to access neurosurgical care. Hence, treatment and follow-ups of "shunted" patients are difficult. In this context, endoscopic third ventriculostomy with choroid plexus cauterization (ETV/CPC) provides an opportunity for an affordable and less constraining treatment for hydrocephalus children under 12 months of age. METHODS: We performed a retrospective analysis of ETV/CPC performed on infants from July 2013 to January 2015. Patients were followed postoperatively on day 15, month 6, and month 12. Statistical analysis was conducted using Prism 9 GraphPad software. ETV successes were categorized according to the patient's age into 3 groups: ≤3 months, 3-6 months, and 6-12 months. Statistical significance was defined at p < 0.05. RESULTS: During the study period, 199 patients were included with 40% of patients aged between 0 and 6 months. The head circumference ranged from 35 cm to 79 cm. The etiology was congenital malformation in 55%. ETV/CPC was a success in 69% of 6- to 12-month-old patients, 54% in the 3- to 6-month-old patients, and 29% in ≤3-month-old patients. Overall, 94 (47%) patients were successfully treated without a shunt. The postoperative infection rate was 1% and mortality at 12 months was 8%. CONCLUSION: In a low-income environment such as Mali, ETV/CPC stands as a viable and alternative treatment option for pediatric hydrocephalus patients; our findings suggest that age is an important factor in predicting ETV success.

Hereditary spastic paraplegia in Mali: epidemiological and clinical features.

BACKGROUND AND PURPOSE: Hereditary spastic paraplegia (HSP) is a group of neurodegenerative diseases divided into pure and complex forms, with spasticity in lower limbs only, or associated with other neurologic and non-neurologic manifestations, respectively. Although widely reported in other populations, very little data exist in sub-Saharan Africa. METHODS: Patients with neurodegenerative features were evaluated over a 19-month period at the Department of Neurology, Teaching Hospital of Point "G", Bamako, Mali. The diagnosis of HSP was considered based on family history and the absence of other known non-genetic causes. Genetic analysis including candidate gene and whole exome sequencing was performed and variant pathogenicity was tested using prediction tools and ACMG guidelines. RESULTS: Of the 170 families with hereditary neurological disorders enrolled, 16 had features consistent with HSP, a frequency of 9%. The average age of onset was 14.7 years with 46% starting before age 6. The male/female ratio was 2.6:1. Complex forms were seen in 75% of cases, and pure forms in 25%. Pyramidal findings were present in all patients. Associated features included mental retardation, peripheral neuropathy, epilepsy, oculomotor impairment and urinary urgency. Most patients were treated with a muscle relaxant and physical therapy, and restorative surgery was done in one. Genetic testing identified novel variants in three families (19%). CONCLUSION: This study confirms the clinical variability of HSPs and adds African data to the current literature.

Morbidity and Mortality of Typhoid Intestinal Perforation Among Children in Sub-Saharan Africa 1995-2019: A Scoping Review.

BACKGROUND: Typhoid fever incidence and complications, including intestinal perforation, have declined significantly in high-income countries, with mortality rates <1%. However, an estimated 10.9 million cases still occur annually, most in low- and middle-income countries. With the availability of a new typhoid conjugate vaccine licensed for children and recommended by the World Health Organization, understanding severe complications, including associated mortality rates, is essential to inform country-level decisions on introduction of this vaccine. This scoping review summarizes over 20 years of the literature on typhoid intestinal perforation in sub-Saharan Africa. METHODS: We searched EMBASE, PubMed, Medline, and Cochrane databases for studies reporting mortality rates due to typhoid intestinal perforation in children, under 18 years old, in sub-Saharan Africa published from January 1995 through June 2019. RESULTS: Twenty-four papers from six countries were included. Reported mortality rates ranged from 4.6-75%, with 16 of the 24 studies between 11 and 30%. Thirteen papers included postoperative morbidity rates, ranging from 16-100%. The most documented complications included surgical site infections, intra-abdominal abscesses, and enterocutaneous fistulas. High mortality rates can be attributed to late presentation to tertiary centers, sepsis and electrolyte abnormalities requiring preoperative resuscitation, prolonged perforation-to-surgery interval, and lack of access to critical care or an intensive care unit postoperatively. CONCLUSIONS: Current estimates of mortality related to typhoid intestinal perforation among children in sub-Saharan Africa remain unacceptably high. Prevention of typhoid fever is essential to reduce mortality, with the ultimate goal of a comprehensive approach that utilizes vaccination, improvements in water, sanitation, and hygiene, and greater access to surgical care.

The impact of African-trained neurosurgeons on sub-Saharan Africa.

OBJECTIVE: Sub-Saharan Africa (SSA) represents 17% of the world's land, 14% of the population, and 1% of the gross domestic product. Previous reports have indicated that 81/500 African neurosurgeons (16.2%) worked in SSA-i.e., 1 neurosurgeon per 6 million inhabitants. Over the past decades, efforts have been made to improve neurosurgery availability in SSA. In this study, the authors provide an update by means of the polling of neurosurgeons who trained in North Africa and went back to practice in SSA. METHODS: Neurosurgeons who had full training at the World Federation of Neurosurgical Societies (WFNS) Rabat Training Center (RTC) over the past 16 years were polled with an 18-question survey focused on demographics, practice/case types, and operating room equipment availability. RESULTS: Data collected from all 21 (100%) WFNS RTC graduates showed that all neurosurgeons returned to work to SSA in 12 different countries, 90% working in low-income and 10% in lower-middle-income countries, defined by the World Bank as a Gross National Income per capita of ≤ US$995 and US$996-$3895, respectively. The cumulative population in the geographical areas in which they practice is 267 million, with a total of 102 neurosurgeons reported, resulting in 1 neurosurgeon per 2.62 million inhabitants. Upon return to SSA, WFNS RTC graduates were employed in public/private hospitals (62%), military hospitals (14.3%), academic centers (14.3%), and private practice (9.5%). The majority reported an even split between spine and cranial and between trauma and elective; 71% performed between 50 and more than 100 neurosurgical procedures/year. Equipment available varied across the cohort. A CT scanner was available to 86%, MRI to 38%, surgical microscope to 33%, endoscope to 19.1%, and neuronavigation to 0%. Three (14.3%) neurosurgeons had access to none of the above. CONCLUSIONS: Neurosurgery availability in SSA has significantly improved over the past decade thanks to the dedication of senior African neurosurgeons, organizations, and volunteers who believed in forming the new neurosurgery generation in the same continent where they practice. Challenges include limited resources and the need to continue expanding efforts in local neurosurgery training and continuing medical education. Focus on affordable and low-maintenance technology is needed.

Epidemiology of human dermatophytoses in Africa.

In this critical literature review, we summarize the epidemiological trends of dermatophytoses reported in Africa. Our findings clearly emphasize the heavy burden of dermatophytosis in Africa. Tinea capitis is the primary clinical presentation of dermatophytosis in African children throughout the entire African continent. The disease affects more than 20% of school-age children in West Africa, while the prevalence ranges from 10% to more than 70% in other regions of Africa. In African adults, the presence of tinea corporis is the most frequent indicator of dermatophytosis. However, epidemiological studies have been primarily conducted on particular patient groups that are not representative of the general population. We examined dermatophyte species distribution patterns. We observed a predominance of anthropophilic dermatophytes, mainly T. violaceum, in the North and East of Africa and both T. soudanense and M. audouinii in the Western and Central regions of the continent. Interestingly, the zoophilic species, M. canis, has recently emerged in North and East Africa. Optimization of both mycology diagnosis capacities and epidemiological methodology would provide insight into the role that climate and other global aspects of the human environment play in dermatophyte epidemiology. We advocate that using a multisectoral and collaborative strategy would strengthen such future studies.

Dermatophytosis among Schoolchildren in Three Eco-climatic Zones of Mali.

BACKGROUND: Dermatophytosis, and particularly the subtype tinea capitis, is common among African children; however, the risk factors associated with this condition are poorly understood. To describe the epidemiology of dermatophytosis in distinct eco-climatic zones, three cross-sectional surveys were conducted in public primary schools located in the Sahelian, Sudanian and Sudano-Guinean eco-climatic zones in Mali. PRINCIPAL FINDINGS: Among 590 children (average age 9.7 years) the overall clinical prevalence of tinea capitis was 39.3%. Tinea capitis prevalence was 59.5% in the Sudano-Guinean zone, 41.6% in the Sudanian zone and 17% in the Sahelian eco-climatic zone. Microsporum audouinii was isolated primarily from large and/or microsporic lesions. Trichophyton soudanense was primarily isolated from trichophytic lesions. Based on the multivariate analysis, tinea capitis was independently associated with male gender (OR = 2.51, 95%CI [1.74-3.61], P<10(-4)) and residing in the Sudano-Guinean eco-climatic zone (OR = 7.45, 95%CI [4.63-11.99], P<10(-4)). Two anthropophilic dermatophytes species, Trichophyton soudanense and Microsporum audouinii, were the most frequent species associated with tinea capitis among primary schoolchildren in Mali. CONCLUSIONS: Tinea capitis risk increased with increasing climate humidity in this relatively homogenous schoolchild population in Mali, which suggests a significant role of climatic factors in the epidemiology of dermatophytosis.

A double-blind randomized placebo-controlled clinical trial of squalamine ointment for tinea capitis treatment.

BACKGROUND: Novel treatments against for tinea capitis are needed, and the natural aminosterol squalamine is a potential topical antidermatophyte drug candidate. OBJECTIVES: This phase II randomized double-blind placebo-controlled clinical trial aimed at testing the efficacy and safety of a three-week squalamine ointment regimen for the treatment of tinea capitis. PATIENTS: Males aged 6-15 years presenting with tinea capitis were treated with either topical squalamine ointment or placebo for 3 weeks. The primary endpoint was complete clinical cure. The secondary endpoints were the occurrence of local and/or systemic adverse events, mycological cure, and partial clinical response. Prospective follow-up of clinical adverse events was performed daily. RESULTS: Five patients were treated with 1% squalamine ointment and 15 with placebo. No complete cure was observed. No clinical or biological adverse event was recorded. A significantly (p = 0.03) better hair-growth score, indicating a partial clinical improvement of the tinea capitis lesion, was observed in the patients treated with squalamine compared to those treated with placebo. CONCLUSION: This three-week squalamine ointment regimen was well tolerated and showed an encouraging partial clinical activity for the treatment of tinea capitis. Further studies are needed to evaluate the efficacy of topical squalamine alone against tinea corporis or in combination with a systemic antidermatophyte drug against tinea capitis.

[Knowledge and practices of traditionnal birth attendant on birth asphyxia in Kolokani (Mali)]. / Connaissance et pratique des accoucheuses traditionnelles sur l'asphyxie du nouveau-ne dans le district sanitaire de Kolokani.

INTRODUCTION: In Mali, according to the DHS IV in 2006, the neonatal mortality rate is 46 per thousand live births and is higher in rural areas (61‰) than in urban areas (45‰). The proportion of home births is 54%.The aim of our study was to assess the knowledge and practices of traditional birth attendants (TBAs) on pregnancy and newborn asphyxia in the health district of Kolokani. METHODOLOGY: It was a descriptive cross-sectional study conducted in the health district Kolokani 1st September to 04 October 2004. The study population consisted of traditional birth attendants TBAs. Our study covered 10% of the villages is Kolokani 28 villages randomly selected in 22 health areas. A total of 43 TBAs were interviewed. Data were entered with software EPI-Info version 6 and analyzed on SPSS11. RESULTS: TBAs have said that the main pathologies compromising reproductive health were toxoplasmosis (79%), pelvic pain (74.4%), malaria (46.5%). The main diseases were heralds of a difficult delivery were toxoplasmosis (16.2%), pelvic pain (16.2%), polyhydramnios (11.6%), but 67.4% of TBAs unaware of these pathologies.The main actions carried out before and during childbirth were displays of loin cloth (48.8%), floor sweeping (34.8%), display of drawsheet (30.2%), incantations (27.9%), the knotted strings "tafo" (32.5%). Deliveries conducted by TBAs have been estimated at 52% for all areas. CONCLUSION: At the end of our study we recommend TBA training to reduce the risks associated with home births.

INTRODUCTION: Au Mali, selon l'EDSM IV de 2006, le taux de mortalité néonatale est de 46 pour mille naissances vivantes et est plus élevé en zone rurale (61‰)qu'en zone urbaine(45‰).La proportion d'accouchement à domicile est de 54%. L'objectif de notre étude était d'évaluer les connaissances etles pratiques des accoucheuses traditionnelles sur la grossesse et l'asphyxie du nouveau-né dans le district sanitaire de Kolokani. MÉTHODOLOGIE: Il s'agissait d'une étude transversale descriptivemenée dans le district sanitaire de Kolokani du 1er septembre au 04 Octobre 2004. La population d'étude était constituée par les accoucheuses traditionnelles (AT). Notre étude couvrait 10% des villages de Kolokani soit 28 villages sélectionnés de façon aléatoire dans les 22 aires de santé. Au total 43 AT ont étéinterrogées.Les données ont été saisies sur EPI-Info version 6fr, analysé sur SPSS11. RÉSULTATS: Les AT ont déclaré que les principales pathologies compromettant la santé de la reproduction étaient la toxoplasmose (79%), la douleur pelvienne (74,4%), le paludisme (46,5%). Les principales pathologies annonciatrices d'accouchement difficile étaient la toxoplasmose (16,2%), la douleur pelvienne (16,2%), l'hydramnios (11,6%) et par contre 67,4% des AT ignoraient ces pathologies. Les principaux gestes effectués avant et pendant l'accouchement ont été l'étalage de pagne (48,8%), le balayage de sol (34,8%), l'étalage de l'alèze (30,2%), les incantations (27,9%), les cordelettes à nœuds « tafo ¼ (32,5 %).Les accouchements effectués par les AT on été estimés à 52% pour l'ensemble des aires. CONCLUSION: Au terme de notre étude nous recommandons la formation des AT en vue de réduire les risques liés aux accouchements à domicile.

A MALDI-TOF MS procedure for clinical dermatophyte species identification in the routine laboratory.

The conventional identification of dermatophytes requires a long turnaround time and highly skilled mycologists. We have recently developed a tandardized matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) assay to routinely identify molds of potential clinical significance. This study objective was to determine if this same assay could also be employed to identify clinical dermatophytes in the routine laboratory setting. The effects of the inclusion of cycloheximide in the culture medium and incubation time were tested after building a reference spectra library that included 48 well-characterized isolates of 17 dermatophyte species. Then these same isolates were prospectively identified using this library. MALDI-TOF MS-based identification was effective regardless of the presence of cycloheximide or incubation time as 130/133 (97.8%) of the clinical isolates were appropriately identified. Two Microsporum canis isolates yielded uninformative spectra and one M. audouinii isolate was misidentified. Since one only requires a small colony for MALDI-TOF MS analysis, accurate identifications were obtained in 3-6 days and, specifically, before the appearance of their characteristic morphological features. Consequently, identification turnaround time was dramatically reduced as compared to that needed for conventional morphological identification. In conclusion, this standardized MALDI-TOF MS-based identification procedure for filamentous fungi effectively identifies clinical dermatophyte isolates and drastically reduces the response times in the routine clinical laboratory.

In vitro activity of aminosterols against dermatophytes.

We recently reported that aminosterols are fungicidal due to their disrupting the outer membranes of yeasts and that they have a significant in vitro activity against various mould species. Yet, their activity against dermatophytes had never been tested. This study's objective was to evaluate the in vitro activity of squalamine and a synthetic aminosterol derivative (ASD) against various dermatophytes. Susceptibility testing of squalamine, ASD, terbinafine, and griseofulvin was performed, in triplicate, in accord with the Clinical Laboratory and Standards Institute's M38-A2 procedure, using an 80% growth inhibition endpoint. The studies included the following dermatophytes: Trichophyton rubrum, T. mentagrophytes, T. soudanense, Microsporum canis, M. audouinii, M. persicolor; M. cookie and M. gypseum. Squalamine and ASD showed significant in vitro activity against these dermatophytes. The minimum inhibitory concentrations (MICs) ranged from 4-16 mg/l and from 2-8 mg/l for squalamine and ASD, respectively. These findings support further clinical studies of aminosterols activity against superficial dermatophyte infections.

Pseudallescheria/Scedosporium complex species identification by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry.

Because timely and accurate identification of members of the Pseudallescheria/ Scedosporium species complex (PSC) is clinically relevant, the objective of this investigation was to study the stability and influence of the main variable factors in the routine clinical laboratory to the potential use the Matrix-Assisted Laser Desorption Ionization-Time-Of-Flight (MALDI-TOF MS) in the identification of these fungi. Twenty-two PSC reference strains, three clinical isolates, an αHCCA matrix, and an Autoflex I spectrometer with BioTyper software (Bruker) were employed in this study. Intra-and inter-specimen composite correlation indices for each MS spectrum as compared to a reference spectrum were computed. MS identification was stable after the fungi were subcultured over a 1-month period. While neither culture medium (Sabouraud vs. Malt extract) nor protein extraction methods (formic acid vs. trifluoroacetic acid) significantly influenced the quality of the MS identifications, they were considerably increased from day 3 to day 6 of incubation. MALDI-TOF MS can be used in the routine clinical laboratory in the identification of members of the complex provided that valid spectra libraries are developed. Although preliminary results are encouraging, further studies are warranted to demonstrate whether MS can distinguish the species that have recently been described using multilocus sequence analysis within P. boydii sl. and to validate its use in the routine clinical laboratory for identifying clinically relevant moulds.

Molecular markers of resistance to sulphadoxine-pyrimethamine one year after implementation of intermittent preventive treatment of malaria in infants in Mali.

BACKGROUND: Intermittent preventive treatment in infants (IPTi) with sulphadoxine-pyrimethamine (SP) given during routine vaccinations is efficacious in preventing malaria disease and shows no interaction with the vaccines. However, there is a fear that IPTi may result in a rapid increase of parasite resistance to SP. METHODS: To evaluate the impact of IPTi on SP-resistance point mutations, the 22 health sub-districts in the district of Kolokani, Mali, were randomized in a 1:1 ratio and starting in December 2006, IPTi with SP was implemented in 11 health sub-districts (intervention zone), while the other 11 health sub-districts served as the control (non-intervention zone). Blood smears and blood dots on filter paper were obtained from children aged 0-5 years, randomly selected in each of heath sub-districts during two cross-sectional surveys. The first survey was conducted in May 2007 before the start of the transmission season to collect baseline prevalence of the molecular markers of resistance to SP and the second in December 2007 after the end of the transmission season and one year after implementation of IPTi. A total of 427 and 923 randomly selected blood samples from the first and second surveys respectively were analysed by PCR for dhfr and dhps mutations. RESULTS: Each of the three dhfr mutations at codons 51, 59 and 108 was present in 35% and 57% of the samples during the two surveys with no significant differences between the two zones. Dhps mutations at codons 437 and 540 were present respectively in about 20% and 1% of the children during the two surveys in both zones at similar proportion. The prevalence of quadruple mutants (triple dhfr-mutants + dhps-437G) associated with in-vivo resistance to SP in Mali after one year implementation of IPTi was also similar between the two zones (11.6% versus 11.2%, p = 0.90) and to those obtained at baseline survey (10.3% versus 8.1%). CONCLUSION: This study shows no increase in the frequency of molecular markers of SP resistance in areas where IPTi with SP was implemented for one year.