Low-cost liquid medium for in vitro cultivation of Leishmania parasites in low-income countries

Background: Cutaneous Leishmaniasis (CL) induced by Leishmania aethiopica has two clinical manifestations: ulcerating, self-healing CL and non-ulcerating, non-healing CL. The grossly disfiguring multiple nodules on the face and exterior surface of limbs during non-ulcerative CL are sometimes misdiagnosed as other skin infections. Thus the need for definitive and prompt laboratory diagnosis will be required. Identifying Leishmania parasite by culture method is considered as a definitive method for initiation of treatment and as an effective component of leishmaniasis control methods. Recently the involvement of Fas (CD95) and Tumor Necrosis Factor (TNF) Related Apoptosis Inducing Ligand (TRAIL) induced apoptotic pathways were proposed to be involved in tissue destruction and ulceration during L. major induced CL. Aims: 1) to develop an alternative culture media that could minimize the cost for culturing Leishmania from patient lesions. 2) to investigate if the expression of FasL and TRAIL differs in ulcerating and non- ulcerative CL. Methods: GALF-1 media was formulated in our lab and compared to RPMI 1640 medium and conventional Locke s semi solid media (LSSM) which is one of the modifications of Novy-MacNeal-Nicolle (NNN) culture media. Amastigotes transformation, cryopreservation, recovery of parasites, cost and mass cultivation were analysed. Expression of Fas ligand (FasL), TRAIL and apoptosis were assessed by immunohistology in human skin biopsies from L. aethiopica induced ulcerative or non-ulcerative CL. FasL and TRAIL blocking experiments were performed in a murine model of CL. Results and discussion: GALF-1 is cheap and its ingredients available in a low income country such as Ethiopia. GALF-1 was able to transform amastigotes from Ethiopian patients samples and could be used to cultivate promastigotes in large quantities. Cost analysis showed 80% to 95 % decreased costs as compared to conventional media. Promastigotes cultured with GALF-1 could be cryopreserved in liquid nitrogen with comparable re-culture potential to conventional media. Affordability of diagnostic assays is a key issue for resource poor countries and the possibility to cut the cost of the efficient culture method for diagnosis through the use of inexpensive local formulated reagents could improve the diagnosis of leishmaniasis in low income endemic countries. More FasL expressing cells were detected in dermis of ulcerative CL as compared to non-ulcerative CL and controls. TRAIL expression was higher in ulcerative CL as compared to non-ulcerative CL and controls in both epidermis and dermis. Increased dermal expression of FasL and TRAIL was associated with ulcer formation during CL. This correlated with an inhibition of the ulcerative process in a murine CL model during FasL and TRAIL neutralisation.The mechanisms of the involvement of FasL and TRAIL in ulceration was not elucidated and putative reason(s) for the difference in dysregulation of apoptosis are discussed

Treatment of head lice with dimeticone 4% lotion: comparison of two formulations in a randomised controlled trial in rural Turkey

<p>Abstract</p> <p>Background</p> <p>Dimeticone 4% lotion was shown to be an effective treatment for head louse infestation in two randomised controlled trials in England. It is not affected by insecticide resistance but efficacy obtained (70-75%) was lower than expected. This study was designed to evaluate efficacy of dimeticone 4% lotion in a geographically, socially, and culturally different setting, in rural Turkey and, in order to achieve blinding, it was compared with a potential alternative formulation.</p> <p>Methods</p> <p>Children from two village schools were screened for head lice by detection combing. All infested students and family members could participate, giving access to treatment for the whole community. Two investigator applied treatments were given 7 days apart. Outcome was assessed by detection combing three times between treatments and twice the week following second treatment.</p> <p>Results</p> <p>In the intention to treat group 35/36 treated using dimeticone 4% had no lice after the second treatment but there were two protocol violators giving 91.7% treatment success. The alternative product gave 30/36 (83.3%) treatment success, a difference of 8.4% (95% CI -9.8% to 26.2%). The cure rates per-protocol were 33/34 (97.1%) and 30/35 (85.7%) respectively. We were unable to find any newly emerged louse nymphs on 77.8% of dimeticone 4% treated participants or on 66.7% of those treated with the alternative formulation. No adverse events were identified.</p> <p>Conclusion</p> <p>Our results confirm the efficacy of dimeticone 4% lotion against lice and eggs and we found no detectable difference between this product and dimeticone 4% lotion with nerolidol 2% added. We believe that the high cure rate was related to the lower intensity of infestation in Turkey, together with the level of community engagement, compared with previous studies in the UK.</p> <p>Trial Registration</p> <p>Current Controlled Trials ISRCTN10431107</p

Synthesis and antileishmanial activity of novel pyridinium-hydrazone derivatives

A series of substituted phenylethylidenehydrazinylpyridinium derivatives bearing methyl, ethyl, propyl, and propylphenyl groups on the pyridinium nitrogen were synthesized and evaluated for in vitro antileishmanial activity against Leishmania tropica by using the microdilution method. Among the tested compounds, 3d, 5c, 3b, and 3c were found to be the most active derivatives against the promastigotes of L. tropica (IC50 values are 6.90, 9.92, 11.69 and 12.03 mu M, respectively) and to be more active than reference drug meglumine antimonaite (glucantime) (IC50 value: 20.49 mu M). The derivatives investigated in this study may have the potential to be lead compound against leishmanial infection

Epidemiological profile of onychomycosis in the elderly living in the nursing homes

Introduction: Few data is available about the prevalence and the causative microorganisms of onychomycosis in the elderly. The aim of this study was to determine the prevalence and types of onychomycosis in the elderly people living in the nursing homes and to determine the responsible fungal agents. Methods: Elderly people living in nursing homes were examined for onychomycosis. Nails with color and structure changes were suspected of onychomycosis. Scraping materials were evaluated by direct KOH examination and fungal cultures were performed. Results: Totally, 214 elderly persons (59 female, 155 male) from nursing homes were studied. Onychomycosis was suspected clinically in 102 cases and scraping materials were obtained from 122 nails. Clinical types of onyhomycosis were distal subungual in 87, proximal subungual in 21 and white superficial in 14 cases. Fungal spores were seen in 81 (37.8%) samples (3 hand nails, 78 toenails) with direct KOH examination and fungal culture was positive in 54 (25.2%) (3 hand nails, 51 toenails) of them. Trichophyton rubrum was the most frequently isolated fungus (75.9%) followed by Candida glabrata (12.9%). Conclusions: In the elderly, onychomycosis is more common than the general population. However causative fungi are similar; dermatophytes are the most common causative organisms followed by yeasts. (C) 2013 Elsevier Masson SAS and European Union Geriatric Medicine Society. All rights reserved

Investigation of inducible clindamycin resistance among clinical isolates of staphylococci

The aim of this study was to investigate the inducible clindamycin resistance in 306 strains of staphylococci, isolated from various clinical samples over a period of two years. Inducible clindamycin resistance was investigated by D-test using clindamycin and erythromycin discs as per the CLSI guidelines. Out of 177 Staphylococcus aureus strains 53 (17.3%) were methicilline resistant S. aureus (MRSA) and 124 (40.5 %) were methicillin sensitive S. aureus (MSSA). Among the coagulase negative staphylococci, 121 (39.6%) were methicillin sensitive and 8 (2.6%) were methicillin resistant. Out of 306 clinical isolates 99 (36.4%) showed erythromycin resistance. Among the erythromycin resistant isolates 46 (46.5%) were CoNS of which 8 (15.1%) were MRCoNS and 38 (31.4%) MSCoNS. Out of thirty-one erythromycin resistant isolates 20 (6.5% of the total isolates) belonged to MLSBi phenotype, eleven isolates belonged to MLSBc phenotype. We conclude that it is necessary to perform D-test for detection of inducible clindamycin in staphylococci in routine antibiotic susceptibility testing

The prevalence of intestinal parasites and nasal S. aureus carriage among food handlers

food handlers play a major role in the transmission of food-borne diseases, Nasal Staphylococcus aureus (S. aureus) carriage and intestinal parasitism are important risk factors in contamination. The purpose of the authors' study was to determine the prevalence of intestinal parasites and nasal S. aureus carriage among food handlers in Manisa, Turkey, The authors investigated 8,895 people for nasal S. aureus carriage and intestinal parasites. Nasal swab materials and stool samples were examined, and anal cellophane band method was performed. The authors found that S. aureus was isolated in 69 (0.77%) samples. All S. aureus strains were oxacilline sensitive. Intestinal parasites were found in 784 (8.8%) samples. The most common parasites were Entamoeba histolytica (69.9%) and Giardia intestinalis (24.6%). The authors conclude that food handlers should be screened and treated from time to time and that a periodic program of health education on food safety and hygiene should be given