Expression of apoptosis regulating proteins p53 and bcl‑2 in psoriasis.

Background: Dysfunctional apoptosis has an important role in the development of several skin diseases. Psoriatic keratinocytes possess an enhanced ability to resist apoptosis, which might be one of the key pathogenetic mechanisms in psoriasis. P53 and bcl‑2 are two proteins which control apoptosis. Several studies have evaluated the expression of these two proteins in the psoriatic skin, but the results are controversial. Methods: Fifty‑eight cases of psoriatic skin biopsies were studied, and the grade of p53 and bcl‑2 immunostaining was correlated with the histopathological indices of severity. Results: Bcl‑2 expression in the epidermis strongly correlated with the expression in the basal cells and lymphocytes (P – 0.001 and 0.035). There was no correlation with epidermal hyperplasia or with p53 expression in the three compartments. Bcl‑2 expression in the basal layer correlated with the p53 expression in the epidermis (P – 0.027), basal layer (P – 0.015) and the lymphocytes (P – 0.034). There was a strong correlation among the p53 expression in all the compartments. There was also a weak correlation of the p53 expression in the epidermis with the epidermal hyperplasia (P – 0.042). Conclusions: Bcl‑2 does not appear to play an important role in the apoptotic process in psoriasis. In contrast, it is likely that p53 has a far more important role to play. Mutation analysis of the p53 protein is necessary to evaluate if the protein has mutated or if it is of the wild type.

Expression of tumor necrosis factor-α and nuclear factor-kappaB/RelA and the pathogenesis of psoriasis.

Context: Tumor necrosis factor-α (TNF-α) is an important mediator in the pathogenesis of psoriasis. Nuclear factor-kappaB (NF-κB) is a transcription factor that regulates the activity of the proinfl ammatory genes. Psoriasis is an infl ammatory disease and the role of TNF-α and NF-κB, should be considerable. Aims: We studied the role of TNF-α and NF-κB in psoriasis. Materials and Methods: A total of 61 cases of psoriatic skin biopsies were studies and the grade of TNF-α and NF-κB, staining was correlated with the histopathological indices of severity. Statistical Analysis Used: Pearson’s correlation coeffi cient and Chi-square test. Statistical Package for Social Sciences version 13 was used. Results: The TNF-α immunostain in the cytoplasm of the epidermal cells and basal cells showed a strong inverse correlation with the grade of epidermal hyperplasia (P –0.019 and P –0.009, respectively). The epidermal cytoplasmic positivity and lymphocyte positivity for TNF-α did not correlate with the grade of NF-κB immunostaining in the epidermal cell nuclei, basal cells or lymphocytes. The basal cell cytoplasmic positivity for TNF-α correlated with the grade of NF-κB immunostaining in the nucleus of basal cells at a P – 0.005. There was a strong correlation between the epidermal cytoplasmic TNF-α immunostaining with the lymphocyte immunostaining (P –0.08); however, there was no correlation between the TNF-α expression in the other two locations. Conclusions: The study outlines the relationship between NF-κB and TNF-α and their combined role in the development of the characteristic histopathological changes in psoriasis. We hypothesize that NF-κB is involved in stimulating the release of TNF-α which would account for the characteristic histopathological changes of psoriasis. However, it is likely that NF-κB can act independently of TNF-α also in the pathogenesis of psoriasis.

Is necropsy obsolete - An audit of the clinical autopsy over six decades: A study from Indian sub continent.

Introduction: Several studies have documented a decrease in the autopsy rate. This study was taken up to analyse the cause of mortality, the discrepancies between the ante mortem and post mortem diagnosis and the discrepancies between diagnoses according to the type of the disease over a period of six decades. Materials and Methods: Autopsy reports and medical records were retrospectively analyzed over a 63 year period from 1947 to 2010. Results: In our study, there was a steady increase in the percentage of neoplastic cases from 1947 to 1994 after which there has been a signifi cant drop. The cases dying due to infection has also shown a steady decline over the years until 1994. After 1994, there has been a signifi cant increase in the deaths until 2010 (p < 0.05). Death due to cardiac causes has shown an increase until 1962 which has been followed by a steady decline. There has been a sudden rise in the number of cases dying due to renal causes between 1994 and 2000 (p < 0.05). There has been a statistically signifi cant decrease in the discrepancies between the ante mortem and the post mortem diagnosis over the years. Discussion: This study shows that therapeutic and preventive measures correctly instituted have signifi cantly reduce the mortality, particularly with reference to cardiac and infectious causes. The discrepancy between antemortem and post-mortem diagnosis in 2010 is still very high at 9.30 percent. The autopsy will continue to remain relevant especially in elucidating the molecular cause of disease.

Morphometric study of microvessels, epidermal characteristics and inflammation in psoriasis vulgaris with their correlations.

Background: Vascular proliferation, inflammation and epidermal changes are important features in the pathogenesis of psoriasis. Aims: In this study we attempted an objective evaluation of these parameters using morphometry. Methods: Inflammation, microvessels and epidermal parameters were assessed in 50 newly diagnosed cases of psoriasis vulgaris (between 01 Nov 2008 and 31 Oct 2011) by morphometry. Parameters studied were microvessel density, microvessel caliber, inflammatory cell density in dermis, ratio between inner and outer epidermal length, maximum epidermal thickness, minimum epidermal thickness and difference between maximum epidermal thickness and minimum epidermal thickness. Results: Microvessel caliber showed moderate correlation (r = 0.645) and microvessel density, weak correlation (r = 0.226) with inflammatory cell density in dermis. Both these parameters also showed mild positive correlation with "ratio between inner and outer epidermal length". All parameters except minimum epidermal thickness showed mild positive correlation with inflammatory cell density in dermis. Conclusion: All microvessels and epidermal parameters showed positive correlation with dermal inflammation; and epidermal parameters exhibited positive correlation with micro-vascular dilation. It is likely that inflammation is a key factor in the pathogenesis of psoriasis.

Is high pressure liquid chromatography an effective screening tool for characterization of molecular defects in hemoglobinopathies.

Introduction: Hemoglobinopathies constitute entities that are generated by either abnormal hemoglobin or thalassemias. high pressure liquid chromatography (HPLC) is one of the best methods for screening and detection of various hemoglobinopathies but it has intrinsic interpretive problems. The study was designed to evaluate the different mutations seen in cases of hemoglobinopathies and compare the same with screening tests. Materials and Methods: 68 patients of hemoglobinopathies were screened by HPLC. Mutation studies in the beta globin gene was performed using the polymerase chain reaction (PCR)-based allele-specific Amplification Refractory Mutation System (ARMS). Molecular analysis for the sickle cell mutation was done by standard methods. Results: The IVS 1/5 mutation was the commonest mutation seen and it was seen in 26 (38.23%) of the cases. This was followed by the IVS 1/1, codon 41/42, codon 8/9, del 22 mutation, codon 15 mutation and the -619 bp deletion. No mutation was seen in eight cases. There was a 100% concordance between the sickle cell trait as diagnosed by HPLC and genetic testing. Discussion and Conclusion: Our study underlies the importance of molecular testing in all cases of hemoglobinopathies. Although HPLC is a useful screening tool, molecular testing is very useful in accurately diagnosing the mutations. Molecular testing is especially applicable in cases with an abnormal hemoglobin (HbD, HbE and HbS) because there may be a concomitant inheritance of a beta thalassemia mutation. Molecular testing is the gold standard when it comes to the diagnosis of hemoglobinopathies.

Clonality assessment of lymphoproliferative lesions using the polymerase chain reaction: An analysis of two methods.

Background: Lymphoid malignancies are a heterogeneous group of disorders which may be difficult to differentiate from reactive proliferations even after immunohistochemistry. Polymerase chain reaction (PCR) is believed to be a good adjunct tool for diagnosis. Materials and Methods: We examined 24 cases of neoplastic and non-neoplastic lymphoproliferative lesions in this study and evaluated the PCR as an additional tool in the confirmation of the diagnosis. Two different PCR methodologies were evaluated. Results: In the evaluation of the T-cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was highly significant at a P value of <0.05. In the evaluation of the B-cell PCR, it was seen that the correlation using both the commercial kits and the custom-synthesized primers was not significant using either method (P > 0.05). Conclusions: Both the methods showed an excellent concordance for T-cell γ gene rearrangements, However, the same was not seen in the B-cell receptor rearrangements. This may be because of the small sample size or the inability of consensus V primers to recognize complementary DNA sequences in all of the V segments.

Nuclear factor kappa-B and histopathology of chronic gastritis.

Background: Studies suggest that nuclear factor kappa-B (NFκB) activation may be a critical event in the production of proinflammatory molecules in Helicobacter pylori-associated gastritis. Materials and Methods: This study examines the expression and activity of NFκB in situ in antral biopsies of 42 consecutive patients with immunohistochemical techniques. Results: NFκB was highly expressed in the gastric epithelial cells. The number of cells showing activated NFκB correlated with the activity of gastritis (P < 0.05), a measure of neutrophil influx, whereas no correlation was found with the chronicity of inflammation, a measure of the presence of mononuclear inflammatory cells. There was also a strong inverse association with the presence and grade of atrophy. Conclusion: This correlation is direct evidence of the importance of NFκB dependent signal transduction for neutrophil influx in gastritis. The role of NFκB appears to be only in the initial stages of gastritis, there is no role for the molecule in the development of chronic inflammation or atrophy.

The histopathology of chronic gastritis.

Chronic gastritis is a multifactorial disorder thought to be influenced by bacterial and host genetic factors. Histopathological examination is the mainstay of diagnosis, however features like the presence of Helicobacter pylori are difficult to evaluate on biopsy. We evaluated 120 gastric antral biopsies using the revised Sydney system. The density of the inflammatory infiltrate, H pylori and mast cells were evaluated. It was seen that the presence of H pylori is strongly associated with an acute and a chronic inflammatory infiltrate. The presence of neutrophils on biopsy is strongly associated with the presence of H pylori and with the density and the grade of the chronic inflammatory infiltrate. The chronic inflammatory response is an intermediary between the acute inflammatory process and glandular atrophy. The lymphocytic infiltrate is also a precursor lesion of the lymphoid follicles. The presence of mast cells does not appear to be related to any of the other inflammatory parameters. The presence of one feature is a strong indicator for the presence of other inflammatory features.