On-site verification of the CBEU method using UriSelect4 chromogenic medium at the National Institute of Hygiene (INH) of Lomé.

PURPOSE: The Cytobacteriological Examination of Urine (CBEU) is one of the most requested microbiological analyses in the medical bacteriology laboratory. According to the ISO 15189 standard, Medical Biology Laboratory (MBL) must validate the techniques before their use. It is in this perspective that, within the Medical Bacteriology Laboratory of the INH of Lome, we have carried out the on-site verification of the CBEU method using UriSelect4 chromogenic medium at the INH of Lomé. MATERIAL AND METHODS: The biological material was composed of reference bacterial strains and clinical bacterial strains isolated at INH. Failure Modes Effects and Criticality Analysis (FMECA) with 5 â€‹M model was used to perform risk analysis and performances as repeatability, intermediate fidelity/reproducibility, sensitivity and inter-operator variability were evaluated using references strains and different operators. "Comité Français d'Accréditation" (COFRAC) SH GTA 04 was used as reference. Single-factor ANOVA was used to analyze the data. RESULTS: Operating procedures, Patients preparation, Samples and Preparation of culture media were identified as main critical points with a criticality index of 8, 9, 9 and 12 respectively. The use of uriselect4 to perform ECBU was shown to be repeatable and reproducible. The sensibility of Uriselect4 to detect urinary infection was 100% with negligible inter-operator variability. CONCLUSION: This study performed on-site verification of Uriselect4 at the bacteriology lab of INH of Lome and identified some critical points to master. The overall performance criteria from COFRAC SH GTA 04 were conform.

Systemic T Cell Subsets and Cytokines in Patients With Homozygous Sickle Cell Disease and Asymptomatic Urinary Tract Infections in Togo.

Background: In sickle cell disease (SCD), cytokine expression influences the pivotal pathways that contribute to disease pathogenesis. Additional infection could affect the immune profile of patients with SCD and increase disease mortality. The aim of this study was to investigate the cytokines and T helper cells profile in patients with asymptomatic urinary tract infection and homozygous SCD (HbSS). Methods: From July to September 2018, 22 HbSS subjects were recruited at Centre Hospitalier Universitaire Campus in Lomé, Togo, 12 of whom had urinary tract bacterial infections and 10 of whom were uninfected. Cytokines from plasma were measured by the enzyme-linked immunosorbent assay (ELISA) sandwich method, and immune cell profiles were performed by flow cytometry. The immunogenicity of bacteria-derived antigens isolated from the urine of HbSS subjects with asymptomatic urinary tract infections was studied in a cell culture system, and the induction of the cytokines was measured. Results: The mean age of HbSS subjects with urinary tract infections was 20.33 ± 3.58 years, and the male/female ratio was 0.09 (1:11). HbSS subjects with asymptomatic urinary tract infections had elevated plasma levels of interferon gamma (IFN-γ) and interleukin (IL)-10. CD4+Tbet+IFN-γ+ and CD4+FoxP3+IL-10+ T cell populations were decreased in HbSS subjects with asymptomatic urinary tract infections. Bacterial antigens from HbSS subjects induced the production of IL-10 but not IFN-γ in uninfected volunteer donors (HbAA). Conclusion: Our study demonstrated that patients with SCD and asymptomatic urinary tract infections had elevated IFN-γ and IL-10 levels. This chronic inflammatory condition could be a risk for this group of patients in terms of vaso-occlusive crisis. Systematic cytobacteriologic examination of the urine of HbSS subjects would be of interest.

Molecular characterization of high-risk human papillomavirus (HR-HPV) in women in Lomé, Togo.

BACKGROUND: The causative agent of cervical cancer referred to as Human papillomavirus (HPV) remains a real public health problem. Many countries in West Africa, such as Togo have no data on the high-risk HPV (HR-HPV) infection and genotypes distribution. In order to fill the knowledge gap in the field in Togo, the main objective of this study was to determine the prevalence of pre-cancerous lesions of the cervix and HR-HPV genotypes among Togolese women. METHODS: Samples were collected from 240 women by introducing a swab in the cervix. Then, the screening of precancerous cervical lesions using the visual inspection with acetic acid and lugol (VIA / VIL) was conducted. The HR-HPV genotypes were characterised by real-time multiplex PCR. RESULTS: Out of 240 women recruited, 128 (53.3%) were infected by HR-HPV. The most common genotypes were HPV 56 (22.7%), followed by HPV 51 (20.3%), HPV 31 (19.5%), HPV 52 (18.8%) and HPV 35 (17.2%). The least common genotypes were HPV 33 (2.3%) and HPV 16 (2.3%). Among the women, 1.3% (3/240) were positive to VIA/VIL. CONCLUSION: This study allowed HR-HPV genotypes to be characterised for the first time in Lomé, Togo. This will help in mapping the HR-HPV genotypes in West Africa.

Molecular characterization of high-risk humanpapillomavirus genotypes in women with or without cervical lesions at VIA/VILI in Kara, Togo.

Background: Persistent infection with high-risk (HR) papillomavirus (HPV) genotypes plays a central role in the pathogenesis of invasive cervical cancer. Objectives: This study aimed to determine the prevalence and distribution of HR-HPV among women with or without cervical lesions at VIA/VILI in Togo. Methods: Cervical samples were collected from 238 women with or without cervical lesions at VIA / VILI and[c3] DNA [c4]was extracted and analyzed by real-time multiplex PCR. Logistic regression analysis was used to determined risk factors associated with HPV infection.inPietro Annigoni Biomolecular Research Center (CERBA / LABIOGENE) in Burkina Faso. Results: The age of the women ranged from 17 to 61 years old, and most were married (73.5%). The prevalence of HRHPV was 35.71% and this was higher in the age range 35-39 years. The six most common genotypes were HPV 31 (18.7%), HPV 52 (13.82%), HPV 68 (13.01%), HPV 66 (9.76%), HPV 58 (8.13%) and HPV 56 (8.13%). Genotypes HPV 18 (4.07%) and HPV 16 (0.81%) were less frequent.[c5] Married or living with a partner was associated with HPV infection (OR=2,17, IC [1.20-3.91], p<0,009). Conclusion: This study allowed characterizing for the first time in Togo, HR-HPV genotypes. This will help mapping-HR-HPV genotypes circulating in West Africa.

Garlic-derived bioactive compound S-allylcysteine inhibits cancer progression through diverse molecular mechanisms.

The purpose of this review is to discuss the molecular mechanisms underlying the anticancer properties of S-allylcysteine (SAC). Over the decades, evidence derived from in vitro and in vivo studies has shown that this predominant organosulfur component of aged garlic extract has multiple anticancer properties; hence, some potential mechanisms responsible for the anticarcinogenic action have been suggested. These mechanisms include induction of carcinogen detoxification, inhibition of cell proliferation and growth, mediation of cell cycle arrest, induction of cell death, inhibition of epithelial-mesenchymal transition and cell invasion, suppression of metastasis, and induction of immunomodulation in cancer cells. However, the actions and mechanisms are not comprehensive, and important aspects of the anticancer activities of SAC still need to be explored. In light of the current evidence, more specific studies, specifically clinical and epidemiological, are required to advance the promising use of SAC as a chemopreventive and therapeutic agent in cancer.

THERAPEUTIC POTENTIALS OF ETHANOLIC EXTRACT OF LEAVES OF HOLARRHENA FLORIBUNDA (G. DON) DUR. AND SCHINZ (APOCYNACEAE).

BACKGROUND: Holarrhena floribunda is a plant of wide usage in the Togolese folk medicine. A previous ethnobotanical survey on the latex plants of the Maritime region of the country revealed that this plant was included in several recipes curing malaria and microbial infections. Therefore, this study aimed to seek for the effectiveness of the ethanolic extract of the plant in the treatment of these diseases. METHODS: The antimicrobial test was performed using the agar well-diffusion and the NCCLS broth microdilution methods, while the in vivo antimalarial activity was evaluated following the four-day suppressive test of Peters. The acute toxic effects of the extract were monitored after a single oral dose (5,000 mg/kg body weight) administration in NMRI mice. RESULTS: The results indicated that the ethanolic extract of leaves of H. floribunda was active on Staphylococcus aureus ATCC 29213 and clinical strains of Staphylococcus aureus, Salmonella typhi and Klebsiella pneumoniae with MICs ranging from 0.62 to 1.25 mg/mL. The extract also showed significant parasitaemia suppression in a dose-dependent manner. In the acute toxicity assay, the oral administration of the extract to the mice did not affect the relative weight of vital organs, and there were no signs of toxicity or death during the study period. The LD50 of the tested extract was found to be greater than 5,000 mg/kg, indicating its safety. CONCLUSION: This study demonstrates the antibacterial and antimalarial activities of leaves of H. floribunda and then, supports its medicinal use in the treatment of microbial infections.

Major Polymorphisms of Genes Involved in Homocysteine Metabolism in Malaria Patients in Ouagadougou, Burkina Faso.

This study analyzed the four main polymorphisms of the genes in homocysteine metabolism in malaria patients. Forty-two randomly selected subjects, diagnosed positive for Plasmodium falciparum, were included. The four genotypes were detected by real-time PCR using the MTHFR 677C>T, MTHFR 1298A>C, MTR 2756A>G, and MTRR 66A>G detection kit (Sacace Biotechnologies REF: T01002-96-S). The results revealed frequencies of 90% 677CC, 10% 677CT, and 00% 677TT for MTHFR C677T; 78.6% 1298AA, 19% 1298AC, and 2.4% 1298CC for MTHFR A1298C; 61.9% 2756AA, 33.3% 2756AG, and 4.8% 2756GG for MTR A2756G; and 50% of 66AA, 45% of 66AG, and 5% of 66GG for MTRR A66G. Correlations were found between A2756G MTR genotypes and parasitaemia (P = 0.02), MTRR A66G and hemoglobin genotypes (P = 0.009), and MTHFR A1298C and sex (P = 0.01). This study demonstrated for the first time an association between the A2756G MTR alleles and Plasmodium falciparum malaria in Burkina Faso and gave an overview of the genotypic distribution of the major SNPs influencing the metabolism of homocysteine.

Ethnobotanical Study of Latex Plants in the Maritime Region of Togo.

BACKGROUND: In Togo, a little is known about latex plants of the flora used for medicinal purposes. OBJECTIVE: The aim of this study was to identify the latex plant species and their medicinal uses in the Maritime Region of Togo. MATERIALS AND METHODS: The methodology was based on ethnobotanical semi-structural individual interviews of 220 informants. Quantitative ethnobotanical index was used to analyze the data. RESULTS: A total of 33 latex plants species were recorded, from 12 botanical families and 24 genera. The most represented families were Euphorbiaceae and Moraceae with eight species each. The relative importance (RI) value of each species and the informant consensus factor (ICF) of the ailments categories showed that Pergularia daemia (Forssk.) Chiov. (RI = 2.00) and Euphorbia hirta L. (RI = 1.91) were the most versatile in relation to their uses, and infectious diseases (ICF = 0.922) were the category with the greatest consensus among 17 categories. CONCLUSION: These latex plants of Togolese flora are variously used in traditional medicine and it would be important to undertake further investigations in phytochemistry, pharmacology, and toxicology to validate their uses. Abbreviations Used: UV: use value ICF: informant consensus factor RI: relative importance PP: pharmacological properties attributed to a species for a specific ailments AC: ailment categories treated by a given species.

Antimicrobial, Anti-inflammatory and Antioxidant Activities of Jatropha multifida L. (Euphorbiaceae).

BACKGROUND: Jatropha multifida is used in Togolease folk medicine for the healing of chronic wounds. OBJECTIVE: This study aims to investigate antibacterial, anti-inflammatory and antioxidant activities of the leaves ethanolic extract. MATERIALS AND METHODS: The antimicrobial activity was assayed by National Committee for Clinical Laboratory Standards broth microdilution method on strains of Staphylococcus aureus and Pseudomoas aeruginosa isolated from wounds, whereas the anti-inflammatory activity was performed by carrageenan and histamine induced paw edema method in rat modele. The 2, 2-diphenyl-1picrylhydrazyl (DPPH) free radical scavenging and ferric reducing antioxidant power (FRAP) were used for the antioxidant activity. RESULTS: The antibacterial assay showed an in vitro growth inhibition of P. aeruginosa and S. aureus in dose-dependent manner, with minimum inhibitory concentration values ranging from 2.5 to 3.12 mg/mL for S. aureus and from 6.25 to 12.5 mg/mL for P. aeruginosa. The maximum paw anti-inflammatory effect occurred after 3 and 5 h administration of histamine and carrageenan, respectively. The DPPH radical scavenging and the FRAP assays yielded weak antioxidant activity. CONCLUSION: J. multifida possesses antibacterial and anti-inflammatory activities that could justify the use of the plant for the treatment of wounds in the folk medicine. SUMMARY: Antibacterial on germs isolated from wound, anti-inflammatory and antioxidant activities of Jatropha multifida were assayed by NCCLS broth method, carrageenan and histamine, DPPH and FRAP respectively. The results indicated that Jatropha multifida possesses antibacterial and anti-inflammatory and weak antioxidant activities that could justify its use for the treatment of wounds in the folk medicine.

Molecular diagnosis of the human immunodeficiency, Hepatitis B and C viruses among blood donors in Lomé (Togo) by multiplex real time PCR.

This study aimed to compare the sensitivity of multiplex PCR to ELISA technique in the instantaneous detection of HBV, HCV and HIVin blood samples from donors of the National blood Transfusion Centre in Togo. A total of 440 blood samplesfrom volunteer were collected and tested by ELISA and multiplex PCR for HBV, HCV and HIV detection. Among the 440 volunteer blood donors, 83% were female and 17% were male. Age range of 20-29 years was more represented (73%). Whereas, multiplex PCR detected more cases of HBV than ELISA (50% vs 33%, P=0.0155);ELISA more detected HCV than PCR (34% vs 3%, P<0.0001) and HIV (26% vs 7%, P<0.0001). Confirming these observations our data showed that multiplex PCR was more sensitive in the detection of HBV. The sensitivity of ELISA for the detection of HCV and HIV was elevated compared to multiplex PCR. Multiplex PCR was more specific that ELISA for the detection of HCV and HIV.Interestingly, our data showed that the gender do not influenced the sensitivity of either ELISA or multiplex PCR to detect these viruses. This study showed the limit of both ELISA and multiplex PCR in the detection of HBV, HCV and HIV.

Effects of hydroethanolic extracts of Balanites aegyptiaca (L.) Delile (Balanitaceae) on some resistant pathogens bacteria isolated from wounds.

BACKGROUND: The bark of Balanites aegyptiaca (L.) Delile (Balanitaceae) is widely used in the Togolese folk medicine for the treatment of wounds, malaria and skin diseases. This study aimed to evaluate the antioxidant properties and the activity of the bark extract of this plant against some resistant bacteria isolated from wounds. MATERIALS AND METHODS: The antimicrobial activity was assayed using the NCCLS microdilution while the DPPH free radical scavenging and ferric reducing antioxidant power (FRAP) were used for the antioxidant activity. The susceptibility to conventional antibiotics of bacteria was assessed by an agar disk diffusion method using commercial disk. Phenolics were quantified using spectrophotometric methods. RESULTS: The hydroethanolic extracts of the bark inhibited in vitro the growth of Pseudomonas aeruginosa and Staphylococcus aureus in a dose-dependent manner. Bacteriostatic effects were observed on 15 (33%) and 21 (47%) strains of Staphylococcus aureus and Pseudomonas aeruginosa respectively. The MICs and MBCs against Staphylococcus aureus strains ranged respectively from 2.5 to 12.5 µg/mL and 2.5 to 50 µg/mL. Concerning Pseudomonas aeruginosa, these values were equal (12.5 µg/mL). Multidrug resistance was 95% against six antibiotics for Pseudomonas aeruginosa and 91% against five antibiotics for Staphylococcus aureus. The DPPH radical scavenging and the FRAP assays yielded weak antioxidant activity, closely linked with the total phenolic contents. CONCLUSION: This study indicated that the bark extract of Balanites aegyptiaca possess bioactive compounds implicated in the free radical scavenging and antibacterial activities, justifying the use of the plant in the traditional medicines.

Ethnobotanical study of medicinal plants used in the management of diabetes mellitus and hypertension in the Central Region of Togo.

CONTEXT: The Tem tribe in the Central Region of Togo is a population with an extensive knowledge of medicinal plants. However, little is known about their medical practices, principally the use of plants in the management of diabetes mellitus (DM) and hypertension (HTN). OBJECTIVE: The present study documented the indigenous medicinal plant utilization for the management of DM and HTN in the Togo Central Region. METHODOLOGY: From March to October 2010, 55 traditional healers were interviewed about their knowledge on the use of plants for DM and HTN treatment. RESULTS: The results revealed that 35/55 (63.64%) healers had treated at least one case of DM and/or HTN. They highlighted the use of 64 species belonging to 31 families in the treatment of DM and/or HTN. The most used plants against diabetes were Psidium guajava L. (Myrtaceae), Khaya senegalensis A. Juss. (Meliaceae), Sarcocephalus latifolius (Sm.) E.A. Bruce (Rubiaceae), Annona muricata L. (Annonaceae), Bridelia ferruginea Benth. (Phyllanthaceae), and Securidaca longepedunculata Fresen. (Polygalacae), while Allium sativum L. (Liliaceae) and Parkia biglobosa Benth. (Fabaceae), followed by Khaya senegalensis A. Juss. (Meliaceae), Gardenia ternifolia Schumach. (Rubiaceae), and Persea americana Mill. (Lauraceae) were the most commonly cited as antihypertensive. CONCLUSION: The issue revealed that traditional healers of the above mentioned region have basic knowledge regarding herbal medicine for DM and HTN in comparison with previous published reports. Further pharmacological screening of the identified plants should be conducted to ascertain the effectiveness of these plants.

Enteric parasites prevalence at Saint Camille Medical Centre in Ouagadougou, Burkina Faso.

OBJECTIVE: To assess the prevalence of parasitic infections among patients attending Saint Camille Medical Centre and to estimate co-parasitic infections rates. METHODS: From January to December 2009, stool samples were collected from 11 728 persons, aged from five months to 72 years and suffering from gastroenteritis. After macroscopic description, the stools were examined by light microscopy to search for the presence of parasites. RESULTS: From the 11 728 analyzed stools, 6 154 (52.47%) were infected with at least one parasite. Protozoan frequently encountered were: Giardia intestinalis (43.47%), Entamoeba histolytica/Entamoeba dispar (30.74%) and Trichomonas intestinalis (21.72%), while Hymenolepis nana (2.25%) was the most common helminth. Co-infections occurred in 22.34% cases. Within the multi-infected samples, dual and triple infections accounted for 71.18% and 20.00%, respectively. Giardia intestinalis for protozoan and Hymenolepis nana for helminths were the most implicated co-infections. CONCLUSIONS: This study confirms that intestinal parasites are still a public health problem in Burkina Faso. To reduce the incidence of parasitic infections, it is necessary to promote the education of people so that they practice the rules of individual and collective hygiene.