Ethnodiagnostic Skills of the Digo Community for Malaria: A Lead to Traditional Bioprospecting

Malaria is a major public health problem that is presently complicated by the development of resistance by Plasmodium falciparum to the mainstay drugs. Thus, new drugs with unique structures and mechanism of action are required to treat drug-resistant strains of malaria. Historically, compounds containing a novel structure from natural origin represent a major source for the discovery and development of new drugs for several diseases. This paper presents ethnophytotherapeutic remedies, ethnodiagnostic skills, and related traditional knowledge utilized by the Digo community of the Kenyan Coast to diagnose malaria as a lead to traditional bioprospecting. The current study was carried out in three Digo villages of Diani sub-location between May 2009 and December 2009. Data was collected using semi-structured interviews, and open and close-ended questionnaires. A total of 60 respondents (34 men and 26 women) provided the targeted information. The results show that the indigenous knowledge of Digo community on malaria encompasses not only the symptoms of malaria but also the factors that are responsible for causing malaria, attributes favoring the breeding of mosquitoes and practices employed to guard against mosquito bites or to protect households against malaria. This knowledge is closely in harmony with scientific approaches to the treatment and control of the disease. The Digo community uses 60 medicinal plants distributed in 52 genera and 27 families to treat malaria. The most frequently mentioned symptoms were fever, joint pains, and vomiting while the most frequently mentioned practices employed to guard against mosquito bites and/or to protect households against malaria was burning of herbal plants such as Ocimum suave and ingestion of herbal decoctions and concoctions. The Digo community has abundant ethnodiagnostic skills for malaria which forms the basis of their traditional bioprospecting techniques

Influence of Aspilia pluriseta Schweinf (Asteraceae) on the healing of dermal excision wounds (mouse model) and skin sensitization activity (Guinea pig model)

Background: The skin is highly predisposed to injury because of its direct contact with the environment. The aim of treating of wounds is to both hasten healing, and to minimise the occurrence of perturbations of the healing process. Many plants traditionally used to treat wounds have been proven to support the healing process using scientific models. Aspilia pluriseta has been used by a number of communities in East and Southern Africa to treat wounds. Objectives: This study aimed at testing ethnomedical claims of wound healing activity of A. pluriseta using preclinical models. Methods: Aerial parts of the plant were ground and incorporated into an ointment base (10% and 20% w/w) to evaluate the influence of the plant on the healing of acute excision wounds in mice compared to Silverex Cream® and Simple Ointment (B.P.). The 20% ointment was tested for skin sensitization in guinea pigs. Results: The effects of the plant-based ointments on wound contraction and gross epithelialisation time were less than significantly different from the controls (p≥0.05), but histopathologic examination revealed remarkable epithelialisation and collagen deposition in the wounds treated with the these ointments. The 20% A. pluriseta-based ointment induced moderate allergic contact dermatitis. Key words: Aspilia pluriseta, wound healing, skin sensitization, excision wound mode

Plasmacytoid Dendritic Cells Sequester High Prion Titres at Early Stages of Prion Infection

In most transmissible spongiform encephalopathies prions accumulate in the lymphoreticular system (LRS) long before they are detectable in the central nervous system. While a considerable body of evidence showed that B lymphocytes and follicular dendritic cells play a major role in prion colonization of lymphoid organs, the contribution of various other cell types, including antigen-presenting cells, to the accumulation and the spread of prions in the LRS are not well understood. A comprehensive study to compare prion titers of candidate cell types has not been performed to date, mainly due to limitations in the scope of animal bioassays where prohibitively large numbers of mice would be required to obtain sufficiently accurate data. By taking advantage of quantitative in vitro prion determination and magnetic-activated cell sorting, we studied the kinetics of prion accumulation in various splenic cell types at early stages of prion infection. Robust estimates for infectious titers were obtained by statistical modelling using a generalized linear model. Whilst prions were detectable in B and T lymphocytes and in antigen-presenting cells like dendritic cells and macrophages, highest infectious titers were determined in two cell types that have previously not been associated with prion pathogenesis, plasmacytoid dendritic (pDC) and natural killer (NK) cells. At 30 days after infection, NK cells were more than twice, and pDCs about seven-fold, as infectious as lymphocytes respectively. This result was unexpected since, in accordance to previous reports prion protein, an obligate requirement for prion replication, was undetectable in pDCs. This underscores the importance of prion sequestration and dissemination by antigen-presenting cells which are among the first cells of the immune system to encounter pathogens. We furthermore report the first evidence for a release of prions from lymphocytes and DCs of scrapie-infected mice ex vivo, a process that is associated with a release of exosome-like membrane vesicles

Erythrina abyssinica prevents meningoencephalitis in chronic Trypanosoma brucei brucei mouse model

Human African trypanosomiasis is prevalent in Sub-sahara African countries that lie between 14° North and 29° south of the equator. Sixty million people are at risk of infection. Trypanosoma brucei gambesience occurs in West and Central Africa while Trypanosoma brucei rhodesience occurs in East and Southern Africa. The neurological stage of the disease is characterized by neuroinflammation. About 10% of patients treated with the recommended drug, melarsoprol develop post treatment reactive encephalopathy, which is fatal in 50% of these patients, thus melarsoprol is fatal in 5% of all treated patients. This study was aimed at establishing the potential activity of Erythrina abyssinica in reducing neuroinflammation following infection with Trypanosoma brucei brucei. Swiss white mice were divided into ten groups, two control groups and eight infected groups. Infected mice received either methanol or water extract of Erythrina abyssinica at 12.5, 25, 50 or 100 mg/kg body weight. Parasite counts were monitored in peripheral circulation from the third day post infection up to the end of the study. Brains were processed for histology, immunohistochemistry scanning and transmission electron microscopy. Following infection, trypanosomes were observed in circulation 3 days post-infection, with the parasitaemia occurring in waves. In the cerebrum, typical brain pathology of chronic trypanosomiasis was reproduced. This was exhibited as astrocytosis, perivascular cuffing and infiltration of inflammatory cells into the neuropil. However, mice treated with Erythrina abyssinica water extract exhibited significant reduction in perivascular cuffing, lymphocytic infiltration and astrocytosis in the cerebrum. The methanol extract did not have a significant difference compared to the non-treated group. This study provides evidence of anti-inflammatory properties of Erythrina abyssinica and may support its wide use as a medicinal plant by various communities in Kenya

Host cell responses of Salmonella typhimurium infected human dendritic cells

Live attenuated Salmonella are attractive vaccine candidates for mucosal application because they induce both mucosal immune responses and systematic immune responses. After breaking the epithelium barrier, Salmonella typhimurium is found within dendritic cells (DC) in the Peyer's patches. Although there are abundant data on the interaction of S. typhimurium with murine epithelial cells, macrophages and DC, little is known about its interaction with human DC. Live attenuated S. typhimurium have recently been shown to efficiently infect human DC in vitro and induce production of cytokines. In this study, we have analysed the morphological consequences of infection of human DC by the attenuated S. typhimurium mutant strains designated PhoPc, AroA and SipB and the wild-type strains of the American Type Culture Collection (Manassas, VA, USA), ATCC 14028 and ATCC C53, by electron microscopy at 30 min, 3 h and 24 h after exposure. Our results show that genetic background of the strains profoundly influence DC morphology following infection. The changes included (i) membrane ruffling; (ii) formation of tight or spacious phagosomes; (iii) apoptosis; and (iv) spherical, pedunculated membrane-bound microvesicles that project from the plasma membrane. Despite the fact that membrane ruffling was much more pronounced with the two virulent strains, all mutants were taken up by the DC. The microvesicles were induced by all the attenuated strains, including SipB, which did not induce apoptosis in the host cell. These results suggest that Salmonella is internalized by human DC, inducing morphological changes in the DC that could explain immunogenicity of the attenuated strains

Host cell responses of Salmonella typhimurium

Live attenuated Salmonella are attractive vaccine candidates for mucosal application because they induce both mucosal immune responses and systematic immune responses. After breaking the epithelium barrier, Salmonella typhimurium is found within dendritic cells (DC) in the Peyer's patches. Although there are abundant data on the interaction of S. typhimurium with murine epithelial cells, macrophages and DC, little is known about its interaction with human DC. Live attenuated S. typhimurium have recently been shown to efficiently infect human DC in vitro and induce production of cytokines. In this study, we have analysed the morphological consequences of infection of human DC by the attenuated S. typhimurium mutant strains designated PhoPc, AroA and SipB and the wild-type strains of the American Type Culture Collection (Manassas, VA, USA), ATCC 14028 and ATCC C53, by electron microscopy at 30 min, 3 h and 24 h after exposure. Our results show that genetic background of the strains profoundly influence DC morphology following infection. The changes included (i) membrane ruffling; (ii) formation of tight or spacious phagosomes; (iii) apoptosis; and (iv) spherical, pedunculated membrane-bound microvesicles that project from the plasma membrane. Despite the fact that membrane ruffling was much more pronounced with the two virulent strains, all mutants were taken up by the DC. The microvesicles were induced by all the attenuated strains, including SipB, which did not induce apoptosis in the host cell. These results suggest that Salmonella is internalized by human DC, inducing morphological changes in the DC that could explain immunogenicity of the attenuated strains

Late seroconversion in HIV-resistant Nairobi prostitutes despite pre-existing HIV-specific CD8(+) responses

Resistance to HIV infection in a small group of Kenyan sex workers is associated with CD8(+)-lymphocyte responses to HIV cytotoxic T-lymphocyte (CTL) epitopes. Eleven prostitutes meeting criteria for HIV resistance seroconverted between 1996 and 1999. The occurrence and specificity of preexisting HIV-1 epitope–specific responses were examined using the IFN-γ enzyme–linked immunospot assay, and any epitopes recognized were cloned and sequenced from the infecting viral isolate. Immunologic and behavioral variables were compared between late seroconverters and persistently uninfected sex worker controls. HIV-1 CTL epitope responses were present in four of six cases, 5–18 months before seroconversion, and their presence was confirmed by bulk CTL culture. A possible viral escape mutation was found in one of six epitopes. The key epidemiologic correlate of late seroconversion was a reduction in sex work over the preceding year. In persistently uninfected controls, a break from sex work was associated with a loss of HIV-specific CD8(+) responses. Late seroconversion may occur in HIV-1–resistant sex workers despite preceding HIV-specific CD8(+) responses. Seroconversion generally occurs in the absence of detectable CTL escape mutations and may relate to the waning of HIV-specific CD8(+) responses due to reduced antigenic exposure