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The therapeutic benefits of medicinal plants in terms of anthelmintic properties are known since time immemorial in India, particularly among natives of the Northeast India. However, only sporadic and scarce reports on scientific validation of these plants are available. The present study was conducted on the cestode Raillietina echinobothrida, to establish whether the anthelmintic activity of Potentilla fulgens, Alpinia nigra and Millettia pachycarpa was mediated by apoptosis or not. Light microscopic observation following MTT assay revealed the highest percentage of inhibition of viability among the worms by methanol extract of M. pachycarpa (89.33%), followed by A. nigra (65%) and P. fulgens (37%). Ultrastructural observations revealed swelling of mitochondria, disruption of mitochondrial membrane, vacuolization of mitochondria, appearance of apoptotic bodies in the cytoplasm, disintegration of nuclear membrane and nucleolus were very common throughout the tegument. DAPI stained specimens showed typical morphology of apoptosis, like nuclear condensation and fragmentation in the extracts treated parasites. A decrease in mitochondrial membrane potential was also recorded in the treated groups. Confirmatory TUNEL assay and DNA fragmentation assay of the extracts treated parasites also confirmed the apoptotic nature of cell death and is concluded to be responsible for paralysis and death of the parasite., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Preeclampsia is a unique life-threatening disorder of human pregnancy associated with the abnormal placentation caused by the inadequate trophoblastic invasion due to altered apoptosis of these cells. The aim of the present study was to assess and compare the apoptosis in trophoblastic cells in various zones (villous and extravillous) of placentas of preeclamptic and normotensive nonproteinuric pregnant women., Methods: Hematoxylin eosin staining, TUNEL assay and M30 immunostaining techniques were used for studying apoptosis in trophoblastic cells of placentas of two groups. The results of apoptotic indices by these techniques were compared in preeclamptic group., Results: The TUNEL apoptotic indices were higher in all the zones of placentas of preeclamptic group as compared to control group though the results were not statistically significant. M30 immunostaining also gave higher apoptotic indices in all the zones of preeclamptic placentas as compared to the normal group but the result of apoptotic index of basal plate was not statistically significant. M30 immunostaining was absent in stromal cells., Conclusion: In our study, the trophoblastic apoptotic rates were more in villous as well as extravillous zones of preeclamptic placentas as compared to normal placentas and M30 immunostaining was found to be more sensitive and specific for the apoptotic trophoblastic cells as compared to TUNEL assay., (© 2012 Wiley Periodicals, Inc.)

Angiogenesis is a fundamental process in tumor growth and metastasis. Expression of vascular endothelial growth factor (VEGF) as prognostic indicator has been documented in many types of human tumors. However, the mechanisms responsible for angiogenesis in urinary bladder carcinoma patients are not well defined. Certain carcinogens in tobacco cause DNA damage and may produce specific mutations. In order to investigate the relationship between tobacco smoking, altered patterns of VEGF expression and apoptosis, we have analyzed a group of 125 incident patients with transitional cell carcinoma and 100 cases of control with inflammatory lesions of the bladder. We assessed VEGF overexpression by the use of anti- VEGF antibody through immunohistochemistry, and apoptosis by TUNEL Assay. Expression of VEGF and apoptosis was noticed in 43.2% and 52.8% cases respectively. Both VEGF and apoptosis increased with increasing tumor grade. Apoptosis was seen to be significantly higher in both sexes in the age group of ≥ 50 years (p<0.05) but expression of VEGF was significantly higher among males in the age group of ≥ 50 years (p<0.05). We observed an insignificant association between cigarettes smoking and VEGF overexpression (p>0.05) and significant association with apoptosis. These data support the hypothesis that certain carcinogens derived from cigarette smoking may induce VEGF mutations and apoptosis which in turn are involved in early steps of bladder carcinogenesis.

In the present investigation, the cytotoxic, hydroxyl radical scavenging and topoisomerase inhibition activities of Tabernaemontana divaricata (Apocynaceae) were evaluated. The extracts from leaves of the plant were prepared with different solvents viz. chloroform, methanol, ethyl acetate and hexane. In, in vitro cytotoxicity assay, with cell lines viz HCT-15 (Colon), HT-29 (Colon), 502713 (Colon), MCF-7 (Breast), PC- 3 (Prostrate), it was observed that the ethyl acetate extract was effective against only one colon cell line (502713) at the lowest dose i.e. 10 micro g/ml, whereas the chloroform extract was effective against all the three colon cancer cell lines, at 30 microg/ ml. In order to evaluate the mechanism of cytotoxicity of these extracts, they were assessed for their ability to scavenge hydroxyl radicals in plasmid nicking assay with pBR322. It was observed that all the extracts effectively inhibited the unwinding of supercoiled DNA except hexane extract, which showed the least effect. Since the expression of topo enzymes is linked with cell proliferation so the extracts were also checked for topo I and topo II inhibitory activities. It was noticed that ethyl acetate extract selectively showed inhibition of topo II in topoisomerase II relaxation assay.

Objective: Bcl-2 family of proteins plays a critical role in the regulation of apoptosis. This pathway may be dysregulated leading to an altered ratio of pro- and anti-apoptotic molecules, hence rendering cells resistant to chemotherapy. The objective of this study was to understand the role of Bcl-2 family members in mediation of apoptosis in squamous cell carcinoma of the lung., Results: Bronchoscopically obtained lung biopsies from 30 cases of histologically proven squamous cell carcinoma of the lung in stage III were assessed for the expression of Bcl-2, Bcl-XL, and Bax at the mRNA and protein levels by semi-quantitative reverse transcription (RT-PCR) and immunohistochemistry. The apoptotic index (AI) was determined by the TUNEL assay. The AI ranged from <0.1 to 6.0% with a median of 1.3%. Bcl-2/Bax transcript ratio ranged from 1.5 to 4.5 and Bcl-XL/Bax from 1.3 to 4.0 indicating increased levels of anti-apoptotic molecules at the transcript levels. There was no correlation of the mRNA levels to the apoptotic index. (Wilcoxon-signed rank test.) Immunohistochemistry for proteins revealed that majority of the tumors were Bax predominated. p53 protein immunohistochemical expression was present in 66% cases. The apoptotic index correlated with Bax expression (P < 0.05; Wilcoxon-signed rank test and chi-square test) but not with Bcl-2, Bcl-XL or p53 levels. There was a positive association of p53 with Bax expression., Conclusion: The results of this study indicate that in locally advanced squamous cell carcinoma of the lung, Bax protein is up regulated and determines the level of apoptosis.

Background: Tissue homeostasis and the maintenance of cell populations depend on a delicate balance between the rates of cell proliferation and cell death. Disruption of this balance is an important factor in development and progression of tumors. In the present study we evaluated the growth index in a large group of breast cancer patients and correlated it with various clinical and histopathological features of the tumor., Methods: Estimation of apoptosis was determined by TUNEL assay while immunocytochemistry for proliferating nuclear cell antigen (PCNA) was used as a measure of proliferation. Necrosis was identified morphologically by haematoxylin and eosin staining., Results: A positive correlation was observed between the percentage of PCNA positive cells and the frequency of mitosis (r=0.6117, p<0.0001). A highly statistical significant correlation was observed between type of tissue analyzed and growth index (r=0.46869, p<0.0001). No significant association was observed between hormone receptor status and growth index., Conclusions: The growth index was found to be higher in carcinoma cells that metastasised into lymph nodes compared with primary lesions with no nodal metastasis. Growth index was particularly prominent in high-grade tumors in which increased proliferative activity was evident. Apoptotic cells were detected more frequently in tumor cells with higher rather than lower proliferative activity. This suggests that not only proliferative activity but also capacity for apoptosis is altered in breast tumors.