Profiling of Germline Mutations in Major Hotspot Codons of TP53 Using PCR-RFLP.

Tumor suppressor protein, TP53 also known as the "guardian of the genome" plays a key role in preventing malignant transformation. Almost 50% of human tumors carry mutations in this gene; in the remaining tumors, the TP53 network is functionally inoperative. The majority of TP53 mutations are missense mutations and more than 90% of the missense mutations affect specific codons in the DNA-binding domain, called "hotspot codons." The present study was aimed at analyzing the germline mutation status of four hotspot codons in TP53 namely, codon 175, codon 245, codon 248 (within the DNA binding domain) and codon 72 (outside the DNA binding domain) in cancer cases encountered in a tertiary care hospital in South India by PCR-RFLP. The case-control study included 85-10 subjects respectively. The results of the study indicated that majority of the cancer cases did not harbor germline mutations in the four hot spot codons of TP53. The study further highlights the usefulness of PCR-RFLP as a simple and cost effective tool for checking gene mutations.

Purification, characterization and allergenicity assessment of 26kDa protein, a major allergen from Cicer arietinum.

Chickpea (CP), a legume of the family Fabaceae, is an important nutrient-rich food providing protein, essential amino acids, vitamins, dietary fibre, and minerals. Unfortunately, several IgE-binding proteins in CP have been detected that are responsible for allergic manifestations in sensitized population. Therefore, the prevalence of CP induced allergy prompted us towards purification, characterization and allergenicity assessment of a major ∼26kDa protein from chickpea crude protein extract (CP-CPE). Purification of CP 26kDa protein was done using a combination of fractionation and anion exchange chromatography. This protein was further characterized as "Chain A, crystal structure of a plant albumin" from Cicer arietinum with Mol wt 25.8kDa by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Further, allergenic potential of purified 25.8kDa protein was assessed using in vivo and in vitro model. Purified protein showed IgE-binding capacity with sensitized BALB/c mice and CP allergic patient's sera. Enhanced levels of specific and total IgE, MCP-1, MCPT-1, myeloperoxidase, histamine, prostaglandin D2, and cysteinyl leukotriene were found in sera of mice treated with CP ∼26kDa protein. Further, expressions of Th2 cytokines (i.e. IL-4, IL-5, IL-13), transcription factors (i.e. GATA-3, STAT-6, SOCS-3) and mast cell signaling proteins (Lyn, cFgr, Syk, PLC-γ2, PI-3K, PKC) were also found increased at mRNA and protein levels in the intestines of mice treated with CP ∼26kDa protein. In addition, enhanced release of ß-hexosaminidase, histamine, cysteinyl leukotriene and prostaglandin D2 were observed in RBL2H3 cell line when treated (125µg) with CP 26kDa protein. Conclusively, in vivo and in vitro studies revealed the allergenic potential of purified CP 26kDa protein. Being a potential allergen, plant albumin may play a pivotal role in CP induced allergenicity. Current study will be helpful for better development of therapeutic approaches to prevent the allergenicity in CP sensitized individuals.

An Adult with Haemoptysis: A Rare Case of Congenital Anomaly.

A non-smoker adult male presented with haemoptysis of short duration. Chest radiograph (postero-anterior view) suggested an opaque left hemithorax. Further evaluation of lung lesion pointed towards a left lung hypoplasia with absent left pulmonary artery and a right-sided aortic arch (RAA). Both kidneys were enlarged with multiple cysts and thinning of parenchyma. This case describes a unique coexistence RAA and probable autosomal dominant polycystic kidney disease.

Allergic manifestation by black gram (Vigna mungo) proteins in allergic patients, BALB/c mice and RBL-2H3 cells.

The prevalence of black gram (Vigna mungo) induced allergic reactions are reported from several parts of the world including Asia and Australia. But, a thorough exploration of the allergic reactions induced by black gram proteins is still lacking. Therefore, efforts have been made to explore black gram allergy using in vivo and in vitro approaches. In this study, Simulated Gastric Fluid (SGF) assay and IgE immunoblotting were carried out to identify clinically relevant allergens of black gram. BALB/c mice and RBL-2H3 cells were used for elucidation of allergenic reactions of black gram proteins. Further, this study was extended to screen black gram sensitive patients among nasobronchial allergic patients on the basis of clinical history, skin prick test (SPT), specific IgE levels and IgE immunoblotting. Enhanced levels of specific IgE, IgG1/IgG2a (p < 0.05), histamine (p < 0.05), clinical symptoms, pathological indications in the lungs, intestine and spleen were evident in black gram sensitized BALB/c mice. Moreover, the expression of Th2 cytokine transcripts and GATA-3/T-bet ratio was found enhanced in the treated group. In vitro studies on RBL-2H3 cells,showed increased release of ß-hexosaminidase (p < 0.05), histamine (p < 0.05), cysteinyl leukotriene (p<0.05) and prostaglandin D2 (p < 0.05). Further, 8.5% of screened patients were found allergic to black gram and concomitant sensitization with other allergens has shown the possibility of further enhancement in allergenic problem. Conclusively, the present study suggested that black gram consumption may be responsible for inducing immediate type of allergic sensitization in susceptible subjects.

Factors associated with non-adherence to specific allergen immunotherapy in management of respiratory allergy.

BACKGROUND: Non-adherence to specific allergen immunotherapy is a major hurdle faced by the allergist, contributing to poor clinical outcomes. OBJECTIVES: To assess the independent association of various factors with non-adherence to specific allergen immunotherapy. METHODS: Fifty consecutive (non-adherent) and control (adherent) subjects receiving specific allergen immunotherapy were included in the study and various factors related to non-adherence including socio-demographic, clinical and immunotherapy related variables were compared between the two groups by univariate and multivariate analysis. RESULTS: On univariate analysis, gender, allergic conjunctivitis, family history, progression of disease, perception of immunotherapy, medicine requirement, and the pattern of missed doses greater than two in the last 10, 20 and 30 doses were found to be significantly associated with non-adherence. On multivariate analysis, independent association was observed with allergic conjunctivitis, family history, perception of immunotherapy, missed doses greater than two in the last 10 doses of immunotherapy and medicine requirement. CONCLUSIONS: The independent factors associated with non-adherence may vary between different settings and countries. There is a need for developing individual case holding programmes to improve clinical outcomes in patients receiving specific allergen immunotherapy.