Mapping the Antigenicity of the Parasites in Leishmania donovani Infection by Proteome Serology

BACKGROUND: Leishmaniasis defines a cluster of protozoal diseases with diverse clinical manifestations. The visceral form caused by Leishmania donovani is the most severe. So far, no vaccines exist for visceral leishmaniasis despite indications of naturally developing immunity, and sensitive immunodiagnostics are still at early stages of development. METHODOLOGY/PRINCIPLE FINDINGS: Establishing a proteome-serological methodology, we mapped the antigenicity of the parasites and the specificities of the immune responses in human leishmaniasis. Using 2-dimensional Western blot analyses with sera and parasites isolated from patients in India, we detected immune responses with widely divergent specificities for up to 330 different leishmanial antigens. 68 antigens were assigned to proteins in silver- and fluorochrome-stained gels. The antigenicity of these proteins did not correlate with the expression levels of the proteins. Although some antigens are shared among different parasite isolates, there are extensive differences and no immunodominant antigens, but indications of antigenic drift in the parasites. Six antigens were identified by mass spectrometry. CONCLUSIONS/SIGNIFICANCE: Proteomics-based dissection of the serospecificities of leishmaniasis patients provides a comprehensive inventory of the complexity and interindividual heterogeneity of the host-responses to and variations in the antigenicity of the Leishmania parasites. This information can be instrumental in the development of vaccines and new immune monitoring and diagnostic devices

PFTAIRE Kinase L63 Interactor 1A (Pif1A Protein) Is Required for Actin Cone Movement during Spermatid Individualization in Drosophila melanogaster

A useful model for determining the mechanisms by which actin and actin binding proteins control cellular architecture is the Drosophila melanogaster process of spermatogenesis. During the final step of spermatogenesis, 64 syncytial spermatids individualized as stable actin cones move synchronously down the axonemes and remodel the membranes. To identify new genes involved in spermatid individualization, we screened a collection of Drosophila male-sterile mutants and found that, in the line Z3-5009, actin cones formed near to the spermatid nuclei but failed to move, resulting in failed spermatid individualization. However, we show by phalloidin actin staining, electron microscopy and immunocytochemical localization of several actin binding proteins that the early cones had normal structure. We sequenced the genome of the Z3-5009 line and identified mutations in the PFTAIRE kinase L63 interactor 1A (Pif1A) gene. Quantitative real-time PCR showed that Pif1A transcript abundance was decreased in the mutant, and a transgene expressing Pif1A fused to green fluorescent protein (GFP) was able to fully rescue spermatid individualization and male fertility. Pif1A-GFP localized to the front of actin cones before initiation of movement. We propose that Pif1A plays a pivotal role in directing actin cone movement

DMRT5, DMRT3, and EMX2 Cooperatively Repress at the Pallium-Subpallium Boundary to Maintain Cortical Identity in Dorsal Telencephalic Progenitors

Specification of dorsoventral regional identity in progenitors of the developing telencephalon is a first pivotal step in the development of the cerebral cortex and basal ganglia. Previously, we demonstrated that the two zinc finger doublesex and mab-3 related (Dmrt) genes, Dmrt5 (Dmrta2) and Dmrt3, which are coexpressed in high caudomedial to low rostrolateral gradients in the cerebral cortical primordium, are separately needed for normal formation of the cortical hem, hippocampus, and caudomedial neocortex. We have now addressed the role of Dmrt3 and Dmrt5 in controlling dorsoventral division of the telencephalon in mice of either sex by comparing the phenotypes of single knock-out (KO) with double KO embryos and by misexpressing Dmrt5 in the ventral telencephalon. We find that DMRT3 and DMRT5 act as critical regulators of progenitor cell dorsoventral identity by repressing ventralizing regulators. Early ventral fate transcriptional regulators expressed in the dorsal lateral ganglionic eminence, such as Gsx2, are upregulated in the dorsal telencephalon of Dmrt3;Dmrt5 double KO embryos and downregulated when ventral telencephalic progenitors express ectopic Dmrt5. Conditional overexpression of Dmrt5 throughout the telencephalon produces gene expression and structural defects that are highly consistent with reduced GSX2 activity. Further, Emx2;Dmrt5 double KO embryos show a phenotype similar to Dmrt3;Dmrt5 double KO embryos, and both DMRT3, DMRT5 and the homeobox transcription factor EMX2 bind to a ventral telencephalon-specific enhancer in the Gsx2 locus. Together, our findings uncover cooperative functions of DMRT3, DMRT5, and EMX2 in dividing dorsal from ventral in the telencephalon. SIGNIFICANCE STATEMENT We identified the DMRT3 and DMRT5 zinc finger transcription factors as novel regulators of dorsoventral patterning in the telencephalon. Our data indicate that they have overlapping functions and compensate for one another. The double, but not the single, knock-out produces a dorsal telencephalon that is ventralized, and olfactory bulb tissue takes over most remaining cortex. Conversely, overexpressing Dmrt5 throughout the telencephalon causes expanded expression of dorsal gene determinants and smaller olfactory bulbs. Furthermore, we show that the homeobox transcription factor EMX2 that is coexpressed with DMRT3 and DMRT5 in cortical progenitors cooperates with them to maintain dorsoventral patterning in the telencephalon. Our study suggests that DMRT3/5 function with EMX2 in positioning the pallial-subpallial boundary by antagonizing the ventral homeobox transcription factor GSX2

Studies on The Interaction of leishmania Parasites With Human Chemokines

Since times immemorial, mankind has been plagued by epidemics. They came in different forms and swept through civilization in the form of outbreaks leaving a tale of devastation in their trail, often in catastrophic proportions. The curse of the gods, as epidemics were known in most early civilizations, brought a shudder of fear to every soul from kings to paupers. The curse spared none. However, definitive historical evidence of epidemics is rare. Innumerable references have been made to curses throughout ancient literature from across the globe, though little of these have ever been confirmed. After all, a term as broad as ‘curse’ could mean a lot of other things as well

The Eyes Absent Proteins: Unusual HAD Family Tyrosine Phosphatases

Here, we review the haloacid dehalogenase (HAD) class of protein phosphatases, with a particular emphasis on an unusual group of enzymes, the eyes absent (EYA) family. EYA proteins have the unique distinction of being structurally and mechanistically classified as HAD enzymes, yet, unlike other HAD phosphatases, they are protein tyrosine phosphatases (PTPs). Further, the EYA proteins are unique among the 107 classical PTPs in the human genome because they do not use a Cysteine residue as a nucleophile in the dephosphorylation reaction. We will provide an overview of HAD phosphatase structure-function, describe unique features of the EYA family and their tyrosine phosphatase activity, provide a brief summary of the known substrates and cellular functions of the EYA proteins, and speculate about the evolutionary origins of the EYA family of proteins

Role of chemokines in Leishmania infection

Chemokines are a growing group of chemoattractant cytokines that play important roles in physiological as well as pathological processes. Their roles in various aspects of pathogenesis and inflammation have come to light in the past decade or so. It is becoming increasingly clear that chemokines play a major, perhaps decisive role in Leishmania infections. In this review, we recapitulate important works linking the chemokine system with relation to visceral and cutaneous leishmaniasis over the past decade and attempt to put it all together to propose a single yet unfinished model to account for all the findings

Otoendoscopy - A New Horizon in Ear Surgery

Introduction:-Middle ear surgery is usually performed with an operating microscope. Surgical eradication of cholesteatoma involves canal wall down or intact canal wall techniques. One of the important causes of failure is residual disease in the posterior tympanum specially the sinus tympani. Endoscopic intraoperative visualization of these regions and subsequent disease clearance has been demonstrated to reduce the recurrence of cholesteatoma in mastoidectomy. Objective:-To demonstrate the effectiveness of otoendoscopy along with microscopy vis-a-vis microscopy alone for disease clearance from the sinus tympani. Materials and Methods:-In this prospective study, a total of 40 ears with acquired cholesteatoma were operated upon between January 2012 and August 2013. Twenty three underwent Canal Wall Up (CWU) technique and rest (17) by canal wall down (CWD) procedure. Otoendoscopic evaluation was done after disease clearance of middle ear cleft by microscopy and remaining disease if any cleared. ResuIts:-There were five cases (12.5%) (three CWU mastoidectomy and two CWD mastoidectomy) in which the sinus tympani contained cholestatoma after clearance by microscopy. Otoendoscopy helped to clear remaining cholesteatoma completely in all cases. All patients were followed up for at least six months and no recurrence of cholesteatoma has been detected. So otoendoscopy assisted mastoid surgery provides 100% disease clearance in comparison to 87.5% disease clearance with conventional microsurgery alone. Condusion:-Otoendoscopy provides a panoramic view of the middle ear cavity and should be added to surgical armamentarium during mastoidectomy procedure for better disease clearance and surgical outcome. Keywords:-Otoendoscopy, ear, sinus tympani, cholesteatoma, canal wall up, canal wall down, microscopic surger

PFTAIRE kinase L63 interactor 1A (Pif1A protein) is required for actin cone movement during spermatid individualization in Drosophila melanogaster

A useful model for determining the mechanisms by which actin and actin binding proteins control cellular architecture is th

Evidence of Direct Interactions Between the CC-Chemokines CCL3, CCL4 and CCL5 and Leishmania Promastigotes

Parasites of the genus Leishmania are very successful parasites, possessing multiple host evasive machineries and unique molecules that aid in their survival. It is presumed that a freshly inoculated parasite needs to survive in the hostile extracellular environment for several hours before it is internalized by a macrophage [1,2]. How the parasite survives within this extracellular environment and find their way to a macrophage are matters of great interest and enigma to clinicians and biologists alike. Chemotactic molecules secreted by the parasite, that are to this day uncharacterized, can home certain host cells and probably play a major role in the process of inviting host cells to the site of infection [3]. Host derived chemotactic molecules, especially chemokines also play an important role in the homing of cells to the site of infection [4]. What is not clear is whether these host derived chemotactic factors can also be sensed and utilized by the parasites