Adaptation of a microbead assay for the easy evaluation of traditional anti-sickling medicines: application to DREPANOSTAT and FACA.

CONTEXT: Sickle cell disease is a common inherited blood disorder affecting millions of people worldwide. Due to lack of progress in drug discovery for a suitable treatment, sufferers often turn to traditional medicines that take advantage of the plant extracts activity used by traditional healers. OBJECTIVE: This study optimizes an anti-sickling screening test to identify preparations capable of reverting sickle cells back to the morphology of normal red blood cells. We focused on the miniaturization and practicability of the assay, so that it can be adapted to the laboratory conditions commonly found in less developed countries. MATERIALS AND METHODS: We tested two traditional anti-sickling herbal medicines, FACA® and DREPANOSTAT®, composed of Zanthoxylum zanthoxyloides (Lam.) Zepern. & Timler (Rutaceae) and Calotropis procera (Aiton) Dryand. (Apocynaceae) at screening concentrations of hydroethanol extracts from 0.2 to 1 mg/mL. Potential bioactive molecules present in the extracts were profiled using Ultra High Performance Liquid Chromatography coupled with High Resolution Mass Spectrometry (UHPLC-HRMS/MS) method, identified through HRMS, MS/MS spectra and in silico fragmentation tools. RESULTS: Hydroethanol extracts of FACA® and DREPANOSTAT® showed low anti-sickling activity, inhibiting less than 10% of the sickling process. The UHPLC-HRMS/MS profiles identified 28 compounds (18 in FACA® and 15 in DREPANOSTAT®, including common compounds) among which l-phenylalanine is already described as potential anti-sickling agent. When used as positive control, 7 mg/mL phenylalanine reduced the sickled RBC to 52%. DISCUSSION AND CONCLUSIONS: This assay has been optimized for the easy screening of plant extracts or extracted compounds from bioassay guided fractionation, valuable to laboratories from less developed countries.

Characteristics of Moyamoya Syndrome in Sickle-Cell Disease by Magnetic Resonance Angiography: An Adult-Cohort Study.

Background: Sickle cell disease (SCD) can be complicated by moyamoya syndrome. Brain magnetic resonance angiography (MRA) is a non-invasive method to diagnose this syndrome and, steno-occlusion and moyamoya vessels (MMV) scores have been proposed to evaluate its severity. Previous studies of SCD moyamoya syndrome did not evaluate the severity according to MRA scores. The objective was to assess the characteristics of moyamoya syndrome in an adult cohort of SCD using these MRA scores. Methods: Twenty-five SCD patients with moyamoya syndrome were included using MRA with 3D time of flight technique. We evaluate steno-occlusion score for each hemisphere (range 0-10) from: steno-occlusion severity of internal carotid (ICA) (0-3), anterior cerebral (ACA) (0-3), middle cerebral (MCA) (0-2), and posterior cerebral (PCA) (0-2) arteries. MMV score for each hemisphere (range 0-5) depended from 5 MMV areas: (1) anterior communicating artery (2) basal ganglia (3) ICA/MCA (4) posterior communicating artery/PCA (5) basilar artery. Results: Eight patients (32%) showed unilateral moyamoya syndrome. ICA steno-occlusion was involved in 22 patients (88%), MCA in 23 patients (92%), ACA in 9 patients (36%), and PCA in 3 patients (12%). MMV involved ACoA area in 10 patients (40%), basal ganglia in 13 patients (52%), PCoA/PCA in 10 patients (40%), MCA/ICA in 7 patients (28%), and BA in 1 patient (4%). Steno-occlusion and MMV mean hemisphere scores were 3.4/10 (± 1.42) and 1.6/5 (± 0.71), respectively. Conclusion: Frequent unilateral moyamoya syndrome, uncommon PCA involvement and, moderate steno-occlusion and MMV scores seem to be features of SCD moyamoya syndrome. In future studies, MRA scores could be collected to assess the follow-up in these patients.

A diagnostic protocol designed for determining allergic causes in patients with blood eosinophilia.

BACKGROUND: Blood eosinophilia is a common laboratory abnormality, and its characterization frequently represents a quandary for primary care physicians. Consequently, in France, specialists and particularly hematologists, often must investigate patients who present with blood eosinophilia that often, but not always, occurs because of allergic causes. Both the Departments of Hematology and Parasitology at Toulouse University Hospitals established a collaboration to rule out allergic causes of eosinophilia, particularly helminthiases, prior to initiating more sophisticated investigations. METHODS: Since 2004, the authors employed the same protocol to investigate eosinophilic outpatients who attended the clinic of Parasitology at Toulouse University Hospitals, and they reported the performance of this diagnostic procedure that was designed to be rapid (no hospitalization required) and only moderately expensive. RESULTS: A total of 406 patients who presented with blood eosinophilia greater than 0.5 (×109, giga cells per litter, G/L) had an allergic etiology in 350 (86.2%) cases. Among the remaining 56 subjects, 17 did not undergo a follow-up and 39 were referred to another specialized department, mostly Hematology. However, only 21 patients attended then were subsequently investigated. Non-allergic causes of eosinophilia, including 3 cases of the lymphoid variant of hypereosinophilic syndrome and 2 cases of myeloproliferative disorder, were identified in 14 patients, whereas 7 remained diagnosed as having idiopathic eosinophilia. CONCLUSION: This study underlines the need to investigate patients presenting with even moderate blood eosinophilia. The work-up that was employed appears to be efficient and versatile and may be used by any medical specialist, such as in hematology, infectious disease, or internal medicine departments, who needs to investigate eosinophilic patients and should initially rule out any etiology of allergic eosinophilia.

Young Sprague Dawley rats infected by Plasmodium berghei: A relevant experimental model to study cerebral malaria.

Cerebral malaria (CM) is the most severe manifestation of human malaria yet is still poorly understood. Mouse models have been developed to address the subject. However, their relevance to mimic human pathogenesis is largely debated. Here we study an alternative cerebral malaria model with an experimental Plasmodium berghei Keyberg 173 (K173) infection in Sprague Dawley rats. As in Human, not all infected subjects showed cerebral malaria, with 45% of the rats exhibiting Experimental Cerebral Malaria (ECM) symptoms while the majority (55%) of the remaining rats developed severe anemia and hyperparasitemia (NoECM). These results allow, within the same population, a comparison of the noxious effects of the infection between ECM and severe malaria without ECM. Among the ECM rats, 77.8% died between day 5 and day 12 post-infection, while the remaining rats were spontaneously cured of neurological signs within 24-48 hours. The clinical ECM signs observed were paresis quickly evolving to limb paralysis, global paralysis associated with respiratory distress, and coma. The red blood cell (RBC) count remained normal but a drastic decrease of platelet count and an increase of white blood cell numbers were noted. ECM rats also showed a decrease of glucose and total CO2 levels and an increase of creatinine levels compared to control rats or rats with no ECM. Assessment of the blood-brain barrier revealed loss of integrity, and interestingly histopathological analysis highlighted cyto-adherence and sequestration of infected RBCs in brain vessels from ECM rats only. Overall, this ECM rat model showed numerous clinical and histopathological features similar to Human CM and appears to be a promising model to achieve further understanding the CM pathophysiology in Humans and to evaluate the activity of specific antimalarial drugs in avoiding/limiting cerebral damages from malaria.

[Epidemiological and Clinical features of colorectal cancer at the hematology and oncology ward of Point G in Bamako, Mali, from 2005 to 2011: 113 cases]. / Particularites epidemiologiques et cliniques du cancer colorectal dans le service d'hematologie oncologie medicale du Point G de Bamako au Mali de 2005 a 2011: 113 cas.

AIMS: Epidemiological, clinical and biological characteristics of colorectal cancer vary across continents probably because of different risk factors that are not yet fully listed in countries with limited resources. This study describes the epidemiological and clinical features of colorectal cancer at a University hospital in Bamako, Mali. PATIENTS AND METHODS: A retrospective study that concerned the period from 2005 to 2011 was carried out. It included records of patients with colorectal cancer histologically documented in the service. Were analyzed epidemiological, clinical and biological data stored in files. RESULTS: One hundred-thirteen (113) cases of colorectal cancer were diagnosed representing 15% of all cancers documented over the 7 years. The number of cases increased year by year. The cases of colorectal cancer are not uncommon in young patients (23% under 30 years of age, 60% under 50). The male is significantly more represented in this population of colorectal carriers with a sex ratio M / F = 2. The time between first the symptom and care is long and the diagnosis is made in 85% of cases at a metastatic stage. Localization of the cancer is more frequent in the colon (56%) than in the rectum (44%). Cancer is an adenocarcinoma in 97% of cases. CONCLUSION: These epidemiological and clinical features invite practitioners to reconsider the concept of rarity of colorectal cancer in Africa and vigilance in front of a digestive disorder, even the patient is young, they also emphasize the need to conduct prospective studies to identify specific risk factors and develop appropriate strategies for the prevention and treatment of colorectal cancer in Mali.

BUT: Les caractéristiques épidémiologiques, cliniques et biologiques du cancer colorectal sont variables selon les continents, probablement à cause de facteurs de risque différents et non encore complètement répertoriés dans les pays à faibles moyens. Cette étude décrit les particularités épidémiologiques et cliniques du cancer colorectal dans un service hospitalier de référence spécialisé de Bamako au Mali. PATIENTS ET MÉTHODES: L'étude rétrospective a concerné la période de 2005 à 2011. Elle a inclus les dossiers des patients atteints de cancer colorectal documenté histologiquement. Ont été analysés, les données épidémiologiques, cliniques et biologiques enregistrées dans les dossiers. RÉSULTATS: Cent-treize (113) cas de cancer colorectal ont été colligés et représentaient 15% de l'ensemble des cancers diagnostiqués sur 7 ans. Ces cas étaient en augmentation d'année en année et s'observaient chez une population souvent jeune (23% avant 30 ans, 60% avant 50 ans). Le sexe masculin était significativement plus représenté, soit 2 hommes pour une femme. Le délai entre le premier symptôme et la prise en charge du malade était long avec dans 85% des cas, un diagnostic porté à un stade métastatique. La localisation était plus souvent colique (56%) que rectale (44%). Le cancer était un adénocarcinome dans 97% des cas. CONCLUSION: Ces particularités épidémiologiques et cliniques invitent à reconsidérer le concept de rareté du cancer colorectal en Afrique et à la vigilance de la part des praticiens devant un trouble digestif même chez un sujet jeune, ils soulignent aussi la nécessité de conduire des études prospectives pour identifier des facteurs de risque particuliers et élaborer les stratégies les mieux adaptées pour la prévention et la prise en charge du cancer colorectal au Mali.