Environmental factors associated with the distribution of visceral leishmaniasis in endemic areas of Bangladesh: modeling the ecological niche.

BACKGROUND: Visceral leishmaniasis (VL) is a parasitic infection (also called kala-azar in South Asia) caused by Leishmania donovani that is a considerable threat to public health in the Indian subcontinent, including densely populated Bangladesh. The disease seriously affects the poorest subset of the population in the subcontinent. Despite the fact that the incidence of VL results in significant morbidity and mortality, its environmental determinants are relatively poorly understood, especially in Bangladesh. In this study, we have extracted a number of environmental variables obtained from a range of sources, along with human VL cases collected through several field visits, to model the distribution of disease which may then be used as a surrogate for determining the distribution of Phlebotomus argentipes vector, in hyperendemic and endemic areas of Mymensingh and Gazipur districts in Bangladesh. The analysis was carried out within an ecological niche model (ENM) framework using a maxent to explore the ecological requirements of the disease. RESULTS: The results suggest that VL in the study area can be predicted by precipitation during the warmest quarter of the year, land surface temperature (LST), and normalized difference water index (NDWI). As P. argentipes is the single proven vector of L. donovani in the study area, its distribution could reasonably be determined by the same environmental variables. The analysis further showed that the majority of VL cases were located in mauzas where the estimated probability of the disease occurrence was high. This may reflect the potential distribution of the disease and consequently P. argentipes in the study area. CONCLUSIONS: The results of this study are expected to have important implications, particularly in vector control strategies and management of risk associated with this disease. Public health officials can use the results to prioritize their visits in specific areas. Further, the findings can be used as a baseline to model how the distribution of the disease caused by P. argentipes might change in the event of climatic and environmental changes that resulted from increased anthropogenic activities in Bangladesh and elsewhere.

Single domain antibodies for the knockdown of cytosolic and nuclear proteins.

Single domain antibodies (sdAbs) from camels or sharks comprise only the variable heavy chain domain. Human sdAbs comprise the variable domain of the heavy chain (VH) or light chain (VL) and can be selected from human antibodies. SdAbs are stable, nonaggregating molecules in vitro and in vivo compared to complete antibodies and scFv fragments. They are excellent novel inhibitors of cytosolic/nuclear proteins because they are correctly folded inside the cytosol in contrast to scFv fragments. SdAbs are unique because of their excellent specificity and possibility to target posttranslational modifications such as phosphorylation sites, conformers or interaction regions of proteins that cannot be targeted with genetic knockout techniques and are impossible to knockdown with RNAi. The number of inhibiting cytosolic/nuclear sdAbs is increasing and usage of synthetic single pot single domain antibody libraries will boost the generation of these fascinating molecules without the need of immunization. The most frequently selected antigenic epitopes belong to viral and oncogenic proteins, followed by toxins, proteins of the nervous system as well as plant- and drosophila proteins. It is now possible to select functional sdAbs against virtually every cytosolic/nuclear protein and desired epitope. The development of new endosomal escape protein domains and cell-penetrating peptides for efficient transfection broaden the application of inhibiting sdAbs. Last but not least, the generation of relatively new cell-specific nanoparticles such as polymersomes and polyplexes carrying cytosolic/nuclear sdAb-DNA or -protein will pave the way to apply cytosolic/nuclear sdAbs for inhibition of viral infection and cancer in the clinic.

A disulfide-stabilized human VL single-domain antibody library is a source of soluble and highly thermostable binders.

We have previously shown that incorporation of a second intradomain disulfide linkage into camelid VHH and human VH/VL single-domain antibodies confers increased thermostability. Here, we explored the effects of introducing an additional disulfide linkage, formed between Cys48 and Cys64 (Kabat numbering), into a phage-displayed synthetic human VL library. In comparison to an identical library bearing only the highly conserved Cys23-Cys88 disulfide linkage, the disulfide-stabilized VL library tolerated a similar degree of randomization but retained a higher level of functional diversity after selection with protein L. Both libraries yielded soluble, antigen-specific VLs that recognized a model antigen (maltose-binding protein) with similar affinities, in the micromolar range; however, the disulfide-stabilized antigen-specific VLs were much more thermostable (average ΔTm ∼10°C) than non-disulfide-stabilized VLs. This work provides proof-of-concept for building synthetic antibody libraries using disulfide-constrained immunoglobulin domains, thus avoiding pitfalls of post-hoc disulfide linkage engineering such as impaired antigen binding and reduced expression yield.