Proteomic Analysis of the Human Anterior Pituitary Gland.

The pituitary function is regulated by a complex system involving the hypothalamus and biological networks within the pituitary. Although the hormones secreted from the pituitary have been well studied, comprehensive analyses of the pituitary proteome are limited. Pituitary proteomics is a field of postgenomic research that is crucial to understand human health and pituitary diseases. In this context, we report here a systematic proteomic profiling of human anterior pituitary gland (adenohypophysis) using high-resolution Fourier transform mass spectrometry. A total of 2164 proteins were identified in this study, of which 105 proteins were identified for the first time compared with high-throughput proteomic-based studies from human pituitary glands. In addition, we identified 480 proteins with secretory potential and 187 N-terminally acetylated proteins. These are the first region-specific data that could serve as a vital resource for further investigations on the physiological role of the human anterior pituitary glands and the proteins secreted by them. We anticipate that the identification of previously unknown proteins in the present study will accelerate biomedical research to decipher their role in functioning of the human anterior pituitary gland and associated human diseases.

Human malaria in C57BL/6J mice: an in vivo model for chemotherapy studies.

The present work deals with the development of Plasmodium falciparum stages in mouse model and its potential for the study of efficacy of antimalarial drugs. C57BL/6J mice were infected with multidrug resistant P. falciparum strain then treated with arteether and artesunate. A response was observed to antimalarial drugs in terms of decrease in parasitemia. Mice infected with P. falciparum strain were successfully cured after treatment with either arteether or artesunate. The speed of parasite clearance time and burden of parasitemia differed for each drug and matched the previously reported observations, hence stressing the relevance of the model. These findings thus suggest that P. falciparum infected human RBC (iRBC) - C57BL/6J mice can provide a valuable in vivo system and should be included in the short list of animals that can be used for the evaluation of P. falciparum responses to drugs.

Aggregation-prone near-native intermediate formation during unfolding of a structurally similar nonlenticular ßγ-crystallin domain.

The folding and unfolding of structurally similar proteins belonging to a family have long been a focus of investigation of the structure-(un)folding relationship. Such studies are yet to reach a consensus about whether structurally similar domains follow common or different unfolding pathways. Members of the ßγ-crystallin superfamily, which consists of structurally similar proteins with limited sequence similarity from diverse life forms spanning microbes to mammals, form an appropriate model system for exploring this relationship further. We selected a new member, Crybg3_D3, the third ßγ-crystallin domain of non-lens vertebrate protein Crybg3 from mouse brain. The crystal structure determined at 1.86 Å demonstrates that the ßγ-crystallin domain of Crybg3 resembles more closely the lens ßγ-crystallins than the microbial crystallins do. However, interestingly, this structural cousin follows a quite distinct (un)folding pathway via formation of an intermediate state. The intermediate species is in a nativelike conformation with variation in flexibility and tends to form insoluble aggregates. The individual domains of lens ßγ-crystallins (and microbial homologues) do not follow such an unfolding pattern. Thus, even the closest members of a subfamily within a superfamily do not necessarily follow similar unfolding paths, suggesting the divergence acquired by these domains, which could be observed only by unfolding. Additionally, this study provides insights into the modifications that this domain has undergone during its recruitment into the non-lens tissues in vertebrates.

Monoclonal antibodies to Plasmodium falciparum and their characterization.

An Indian isolate of P. falciparum was propagated in vitro and monoclonal antibodies were raised against this parasite. Of 11 positive hybrids obtained, 3 were cloned and characterized. When tested in IFA with synchronized cultures dominated by mature schizonts, all three monoclonal antibodies showed distinct fluorescent patterns. These three monoclonals reacted with seven of the eight P. falciparum isolates tested. One of these monoclonal antibodies, which showed a fluorescence pattern similar to the merozoite dot fluorescence reported by previous investigators, also inhibited parasite growth in vitro. The antigens that reacted with the monoclonal antibodies have been isolated by using MAb-affinity columns.

Incorporation of radioactive 35SO4(2-) into immunoreactive pituitary lutropin.

The terminal hexosamines of bovine pituitary lutropin are thought to contain a sulfate moiety. In order to test this, a biosynthetic approach was adopted. When rat and buffalo (bovine) pituitaries were incubated with radioactive 35SO4(2-) for 2 h in vitro it was observed that radioactivity test incorporated into trichloroacetic acid-precipitable proteins. When the radioactive proteins were treated with an anti-sheep lutropin serum, radioactivity was found in the immunoprecipitate. The incorporation into rat lutropin like material was very marginal while it was very significant in the case of buffalo lutropin.