Estimating autoimmune hepatitis in patients with hepatic dysfunction.

Autoimmune Hepatitis (AIH) is a chronic inflammatory disease of the liver of unknown aetiology characterized by polyclonal hypergammaglobulinemia and circulating autoantibodies. Objective of the study is to find out the prevalence of autoimmune hepatitis and its causative autoantibody (Anti-nuclear, anti-smooth muscle, anti-liver kidney microsomes-1) following comprehensive diagnostic criteria given by International Autoimmune Hepatitis Group. MATERIAL AND METHODS: 250 patients with raised aminotransferase but, negative for the commonest three clinical conditions viz, viral hepatitis, drug induced liver injury and alcohol induced liver injury were suspected to be a case of autoimmune hepatitis. Recent IAIHG criteria was followed in making diagnosis of autoimmune hepatitis. We did the investigation for viral hepatitis (acute/chronic), drug induced liver injury and alcohol induced liver injury and then measured serum IgG. Indirect Immunofluorescence (IIF) was done for Anti-nuclear antibody and was also done for anti-smooth muscle antibody and anti-liver kidney microsomes-1 antibody simultaneously. RESULT: As per IAIHG criteria, out of 250 patients, Ig "G" was elevated in 88 patients with significant titres for Anti-nuclear antibody (50 patients), anti-smooth muscle antibody (65 patients), anti-liver kidney microsomes-1 antibody (7 patients) and Anti-mitochondrial antibody (18 patients) were found. As per International Autoimmune Hepatitis Group Comprehensive scoring system, probable diagnosis was made in 83 patients (33.2%) and 12 were confirmed for autoimmune hepatitis (4.8%). CONCLUSION: Female preponderance was observed and type 1 Autoimmune Hepatitis was most common whereas type 2 Autoimmune Hepatitis was found in few cases. Most patients had anti-smooth muscle antibody and anti-nuclear antibody positivity indicating the type, but no significant difference was seen in clinical symptoms or biochemical parameters between the different types. 162 patients couldn't be evaluated for autoimmune hepatitis because of serum IgG cut-off criteria >1500 â€‹mg/l.

Factors Affecting Quality of Life of Cancer Patients in North India.

INTRODUCTION: Cancer is the second leading reason of death in many countries, and its burden and prognosis are highly dependent on disease stage at diagnosis. Lack of perception of the population about cancer and its related risk factors usually sequel in defer/delayed presentation and increased treatment morbidity and decrease survival chances. Quality of life (QOL) is defined as Individual view of life, merits, intentions, standards and interests in the framework of culture. AIMS AND OBJECTIVES: To study the socio-demographic profile and factors affecting quality of life of cancer patients. MATERIAL AND METHOD: Using a cross-sectional study, between 1 January 2021 to 31 December 2021, 192 cancer confirmed patients at cancer registry centre at District civil hospital, Ambala cantt. were interviewed using a pre tested semi-structured questionnaire along with EORTC QLQ-C30 (version 3) questionnaire scale for QOL. The data was entered into an excel sheet and was analysed using IBM SPSS version 28. RESULTS: Out of 192 patients, 95 were male and 97 were female. Results shows that female (60.05±19.167) had better mean score of QL2 than male (58.68±18.906) with P value=0.619. Overweight (BMI25.0-29.9) had better mean QL2 score (65.50±18.147) than obese (BMI>30.0) mean QL2 score (61.67±14.803), normal weight (BMI 18.0-24.9) mean QL2 score (59.57±18.575) and underweight (BMI<18.0) mean QL2 score (49.46±19.595) with P value of 0.004. CONCLUSION: Present study found that QOL had significant association with BMI, type of cancer, history of smoking, lost income after cancer diagnosis and stage of cancer at the time of diagnosis. While gender, occupation, age group, religion, residence, marital status, type of family, income, tobacco alcohol and physical activity showed no significance association with QOL.

Male participation in reproductive health care of women and factors associated with interpersonal relationship: A cross-sectional study in a rural community of Ambala District in Haryana.

BACKGROUND: Male participation plays an important role in affecting reproductive health outcomes. Communication between a wife and husband regarding reproductive matters is well recognized as a factor influencing good interpersonal relationships. OBJECTIVES: The objectives of this study were to assess male participation in reproductive health care of women and spousal communications and also to identify factors associated with interpersonal relationship. METHODS: The present descriptive study was conducted among 104 married women aged 18-49 years in a rural community of Haryana during the months of June and July 2019. Study variables included sociodemographic characteristics, reproductive health characteristics, male participation in reproductive health care of women, and interpersonal communication. RESULTS: Male participation was reported by 72 (69.2%) women with an overall involvement of spouses in antenatal care by 71 (68.3%) women. Joint decisions concerning desired number of children, use of contraceptive, and choice of contraceptive use were reported by 47 (45.2%), 48 (46.2%), and 55 (52.9%) women, respectively. Good interpersonal relationship was reported by 80 (76.9%) women and 67 (64.4%) felt their expectations were fulfilled. Male partners played dominant roles in decisions concerning contraceptive use and place of delivery only. Low socioeconomic status came out to be a significant risk factor of not having good interpersonal relationships (P = 0.02). CONCLUSION: Male participation in reproductive health care of women was reasonably high. Spouses were having good interpersonal relationships meeting expectations of women. Efforts should be made for increasing male involvement and encouraging better interpersonal communications for attaining desired reproductive health outcomes for women.

The perspective of our future doctors towards organ donation: a national representative study from India.

INTRODUCTION: Timely donation of organs has helped many get a new life. But in India, the pace of Organ Donation and Transplant (ODT) has been disturbingly sluggish. There is a wide gap between patients who need transplants and the organs that are available in India. Doctors can play a pertinent role in decreasing this gap. Therefore, the present study was done to assess the knowledge, attitude, and perception of the medical students in India regarding Organ Donation (OD). MATERIAL AND METHODS: A cross-sectional study was carried out among the undergraduate students (n=1463) in 10 different medical colleges across the country. Data were collected electronically through Google forms by using a pre-designed, semi-structured, self-administered questionnaire. RESULTS: About 65% of students had a positive attitude regarding ODT. Most of the students acknowledged that Internet sources (57.7%) and print media (46.5%) played a significant role in their awareness. 83% of the students felt that there is a need for effective laws to regulate the process of ODT. It was encouraging that most of the students (69.6%) were not biased for the type of recipients for the organ. CONCLUSION: In the face of acute shortage of donated organs in India, the medical students can be the torchbearers of a positive change. Topics on ODT should be stressed in the initial years and during the early clinical exposure through rapid implementation of the revised Medical curriculum. They can motivate the general public and patients during their routine interactions to make this idea more accessible and acceptable.

Patterns and determinants of healthcare-seeking behavior among hypertensive patients in a rural population of north India: A mixed-method study.

The prevalence of hypertension in india over the last 30 years is known to have increased rapidly. Hypertension is a chronic disease that needs to be managed within its sociological context as experienced by individuals. This study was conducted to identify the quantitative and qualitative determinants of healthcare-seeking behavior among hypertensive individuals in a rural population of north india. METHODOLOGY: A mixed-method study involving 100 adult hypertensive patients was employed. Data were collected on demographic characteristics, health-seeking behavior, and the reasons for choosing a particular health service using a structured questionnaire and interview guide. Descriptive statistics were used to examine the influence of determinants on health-seeking behavior and utilization of healthcare facilities and expressed as percentages. Qualitative data were thematically analyzed. RESULTS: A total of 100 participants with hypertension between 38 and 76 years of age were included in the study. Out of the total study participants, 66.0% of the respondents were males and 34.0% were females. Participants demonstrated a generalized knowledge of hypertension and showed significant gaps and misconceptions in their understanding of the disease and the need for seeking expert care. There was also a lack of sense of trust in the healthcare system. Various demographic, sociocultural, and economic determinants of healthcare-seeking behavior in these rural patients' services were explored. This evidence can prove helpful in streamlining interventional programs and realigning the existing interventions to prevent and control hypertension in rural settings.

A comparative study of sleep quality in different phases of the medical course: A study from Haryana (North India).

BACKGROUND: Inadequate sleep quality is a crucial problem in today's hectic lifestyle. Sleep is known to facilitate cognitive skills, aid memory, and is important for physical, emotional and intellectual health. OBJECTIVES: To determine the sleep quality of the medical undergraduate students and to explore differences in various phases of medical curriculum. MATERIAL AND METHODS: Sleep quality was assessed in 400 medical students in various phases of the medical course using the Pittsburgh Sleep Quality Index (PSQI). Chi square test was used for comparisons of the all seven components of PSQI and sleep quality for different study year, whereas ANOVA was used for comparison of the components and global score. RESULTS: Of the total, 25.3% of the participants classified their sleep quality as either very or fairly bad, and 31.1% reported taking more than 30 min to fall asleep. The average hours slept per night was 7.1 ± 1.21. Of the total, 8.4% of the participants reported using sleep medication at least once a week. Subjective sleep quality and sleep duration were found to be significantly different among the four groups. CONCLUSION: Poor subjective sleep quality was high for students in all class years of the undergraduate medical course. The comparison across the various phases of the course showed that first years reported worse sleep quality than did those in other class years.

Assessment of Quality of Life of the Elderly Living in Rural and Urban Areas of Ambala District: A Comparative Study.

CONTEXT: Aging is a natural process which universally affects all the human beings in the society. As the geriatric population is quiet vulnerable, They might suffer from mental and physical disabilities which consequently threatens their independence. Quality of life among the geriatric population is a global concern as it reflects the status of health and of well being among the set population. AIMS: To assess the quality of life of elderly living in rural and urban areas and compare the role of socio-demographic factors influencing the quality of life of elderly. SETTINGS AND DESIGN: It is a Community based Cross sectional study conducted in urban and rural field practice areas of MMIMSR, Mullana. METHODS AND MATERIAL: Convenience sampling was used. A total of 200 elderly were included in the study. A pretested semi structured questionnaire was used. STATISTICAL ANALYSIS: Data was analysed using SPSS 20.0. RESULTS: According to the sex of the participants, male participants had a higher mean score for QOL as compared to the female participants. Higher mean score was found in each domain for the participants living with their spouses. CONCLUSIONS: The quality of life is better among the individuals who do not suffer from any chronic illness'. The health care services should be strengthened to provide for better healthcare to the elderlies for their morbid conditions.

Knowledge Attitude and Perception of Sex Education among School Going Adolescents in Ambala District, Haryana, India: A Cross-Sectional Study.

INTRODUCTION: Adolescence is a highly dynamic period characterised by rapid growth and development. Adolescents have limited knowledge about sexual and reproduction health, and know little about the natural processes of puberty, sexual health, pregnancy or reproduction. Sex education should be an integral part of the learning process beginning in childhood and continuing into adult life, because it is lifelong process. AIM: This study was carried out to identify the knowledge and attitude of imparting sex education in school going adolescents in rural and urban area of Ambala district. MATERIALS AND METHODS: A cross sectional study design was used to study the knowledge of reproductive and sexual health among school going children. A total of 743 adolescents from age group of 13-19 year were studied, using self designed semi-structured questionnaire to assess the knowledge regarding reproductive and sexual health among adolescents. RESULTS: The mean age of study subjects was 15.958±1.61 years, majority of adolescents i.e., 93.5% favour sex education. An 86.3% said sex education can prevent the occurrence of AIDS and 91.5% of adolescents prefer doctors should give them sex education followed by 83.0% school/teacher and least preference was parents 37.3%. CONCLUSION: There were substantial lacunae in the knowledge about reproductive and sexual health. Students felt that sex education is necessary and should be introduced in the school curriculum.

No Need of Fascia Closure to Reduce Trocar Site Hernia Rate in Laparoscopic Surgery: A Prospective Study of 200 Non-Obese Patients.

BACKGROUND: Laparoscopy is widely practiced and offers realistic benefits over conventional surgery. Port closure is important after a laparoscopic procedure to prevent port site incisional hernia. Larger port size and increasing numbers of ports needed to perform more complex laparoscopic procedures are likely to increase the incidence of port site hernias (PSHs). PSHs tend to develop more frequently at umbilical and midline port sites due to the thinness of the umbilical skin and weaknesses in the linea alba. More than 90% of PSHs occur through 10 mm and large ports can occur through 5 mm ports also. The aim was to study the outcomes and complications in laparoscopic surgery without fascial sheath closure of port site. We compared the results with another group in which fascial closure was done by a standard method. METHODS: This was a prospective study carried out in the Department of Surgery, MMIMSR, Mullana, Ambala, from August 2013 to 2015 in a single unit by a single surgeon. A total of 200 patients were selected randomly for the different laparoscopic procedures. Patients were divided into group A (only skin closure was done without fascia closure) and group B (fascial closure of the port in addition to skin closure). In both groups, we used blunt trocar for the 10 mm port. Skin of the 5 mm port was closed simply. The results in two groups were compared in terms of complications like PSH, bleeding, and wound infection. RESULTS: The outcomes in two groups were compared with and without fascia closure of 10 mm trocar port site. Patients operated for lap cholecystectomy were 170 (85%), 10 (5%) for lap appendicectomy, and 20 (10%) for lap hernia. The study compared the results in two groups mainly for PSH formation. The P value was insignificant and Fischer's exact test result came as 1.00. There were no significant differences between the two groups in terms of PSH, bleeding and infection in non-obese cases. CONCLUSION: In both groups, blunt trocar was introduced into the abdomen. We concluded that this is safe, without visceral injury, and no bleeding was seen in both the groups. We had not encountered any case with PSH formation in follow-up of 6 - 8 months. There was no infection over the port site.

An Epidemiological Study of Malnutrition Among Under Five Children of Rural and Urban Haryana.

INTRODUCTION: A child is future of nation. Malnutrition is a big public health problem in India as it can be attributed for more than half (54 percent) of all under five mortality in India. AIM: To assess prevalence of malnutrition among urban and rural population of Haryana using newly developed WHO growth standards. SETTINGS AND DESIGN: A community based cross-sectional survey was conducted in children of 3-60 months age living in the urban and rural field practice areas of Department of Community Medicine MMIMSR, Mullana, Ambala during January 2012 to December 2012. MATERIALS AND METHODS: Seven hundred and fifty children, aged 3-60 months, were studied for nutritional status, socio-demographic measures were obtained from structured questionnaire and followed by anthropometric assessment using standards methods. Z score for Anthropometric data was calculated by WHO Anthro 2010 software (beta version). STATISTICAL ANALYSIS: Descriptive statistics as well as simple proportion were calculated with SPSS 20. RESULTS: We found that 41.3% children were underweight and 14% were severe underweight. Female children were more nutritionally deprived than males. Among sociodemographic factors maternal educational and working status as well as SES class and rural background of family had greater impact on nutritional status of child. CONCLUSION: We found that almost half of our under five children are underweight, girl child being affected more. For attainment of best possible nutrition and growth in children, targeted short-term strategies addressing underlying risk factors and more long-term poverty alleviation strategies may be needed.

Diagnosis of pulmonary and extrapulmonary tuberculosis based on detection of mycobacterial antigen 85B by immuno-PCR.

We developed a novel indirect sandwich immuno-polymerase chain reaction (I-PCR) assay for the detection of mycobacterial antigen 85B (Ag85B, 30kDa, Rv1886c) in pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB) patients. The amino-modified reporter DNA was covalently attached with the antidetection antibody through a heterobifunctional cross-linking agent succinimidyl 4-[N-maleimidomethyl]-cyclohexane-1-carboxylate. The detection limit of Ag85B by I-PCR was found to be 1 femtogram (fg)/mL, which was 10(6)-fold lower than an analogous enzyme-linked immunosorbent assay (ELISA). The sensitivities of 85% and 77% with I-PCR and 77.6% and 62.5% with ELISA were observed in smear-positive and smear-negative PTB patients, respectively, with high specificity. On the other hand, sensitivities of 84% and 63.7% with I-PCR and 68% and 47.5% with ELISA were observed in confirmed and clinically suspected EPTB cases, respectively, with high specificity.

Women and AIDS: awareness and attitudes in a peri-urban setting in Punjab.

Women account for 39.3% of HIV/AIDS infection in India. There are many biological and socio-cultural factors which make her highly vulnerable to HIV infection. Knowledge regarding HIV/AIDS can help her in protecting herself from the disease. This study was conducted to the level of awareness of young females with respect to knowledge of the HIV/AIDS, its modes of transmission, methods to prevent it, availability of testing and treatment and attitudes in relation to HIV/AIDS. A cross sectional study was done in a peri-urban setting in Patiala, Punjab among 1044 females in the age group 15-24 yrs using a self designed semistructured questionnaire. 93.9% had heard about AIDS. Television was the most common source of information (79.7%). Knowledge about the different modes of transmission was high (68-78%), but knowledge about preventive methods especially condom use was low (34-54%). Increase in educational status was seen to significantly increase the knowledge of the modes of transmission and preventive methods (p < .001). Knowledge of symptoms of HIV was low. (2-15%). Only 28.5% knew about the availability of HIV testing facility. 33.8% knew about treatment available for HIV. The study subjects were having favorable attitude towards people living with HIV/AIDS. There is a need to focus the education programs and the mass media towards preventive methods rather than just the knowledge of the modes of transmission. Greater emphasis needs to be placed on making people aware about the existence of HIV testing facility and availability of treatment.

High Cks1 expression in transgenic and carcinogen-initiated mammary tumors is not always accompanied by reduction in p27Kip1.

Cks1 plays an essential role in SCFSkp2-mediated ubiquitination, and consequently turnover, of the cdk2 inhibitor and tumor supressor p27Kip1. High Cks1 expression is associated with aggressive breast tumors and correlates with low p27Kip1 levels in some cases, although it is also an independent prognostic marker for survival, and provides predictive information in addition to that provided by p27Kip1 alone. In this report we demonstrate that Cks1 protein and mRNA are elevated to very high levels in mammary tumors initiated by erbB2, c-myc and polyoma middle-T (PyMT) in transgenic mice, whereas Cks1 protein is hardly detectable in the normal mammary epithelium. Cks1 is also highly upregulated in rat mammary tumors initiated by methylnitrosourea (MNU). Despite high levels of Cks1 expression, p27Kip1 levels were not reduced, and were in fact slightly higher in mammary tumors initiated by erbB2, PyMT and MNU. In contrast mammary tumors from MMTV-c-myc mice did exhibit low p27Kip1 and higher levels of Skp2. Together, these data suggest that deregulated Cks1 expression might play roles in oncogene and carcinogen-initiated mammary tumorigenesis independent of p27Kip1 turnover in certain tumors. Stable overexpression of Cks1 in human breast carcinoma MCF-7 cells did not significantly reduce p27Kip1 expression, although it conferred resistance to Faslodex (ICI 182780)-mediated inhibition of colony outgrowth in these cells. In contrast, Cks1-depleted MCF-7 cells formed fewer colonies in estrogen-containing medium. Therefore, our studies also suggest that Cks1 levels regulate the responsiveness of ER+ breast cancers to estrogens and anti-estrogens.

Outcome of multi-drug resistant tuberculosis cases treated by individualized regimens at a tertiary level clinic.

AIM: To determine the clinical, radiological and drug resistance profile as well as the factors associated with treatment outcome of Multi-Drug Resistant Tuberculosis (MDR-TB). MATERIAL AND METHODS: All newly diagnosed patients with pulmonary MDR-TB from August 2002 to December 2004 enrolled at New Delhi Tuberculosis Centre, were included in the study. They were followed up clinically, radiologically and bacteriologically by sputum smear, culture and Drug Susceptibility Testing (DST) at regular intervals. According to their DST pattern and previous history of Anti-Tubercular Treatment (ATT), individualized treatment regimens were tailored for each patient. RESULTS: Out of total 27 bacteriologically proven cases of MDR-TB included in this study, 19 were males (mean age and weight 38.5 years and 52.6 kgs, respectively) and eight females (mean age and weight 34.3 years and 40.7 kgs, respectively). A majority (18) were residents of Delhi and the rest hailed from different parts of North India. All of them had a history of previous treatment ranging from six to 34 months. Cavity on chest X-rays was seen in 81%, while 44% showed extensive involvement. The patients received at least four "second line drugs" during their treatment with a mean of 6.2 anti-tubercular drugs during their intensive phase. Of the 27 patients, 13 were cured, 10 defaulted, one died, one is still on treatment and two were referred for surgery. Radiological improvement was observed in two third of cases and chest X-ray of two patients showed a complete resolution. Six predictors were identified for successful outcome of MDR-TB. They include weight gain at six months, culture conversion, radiological improvement during treatment, disease with M. tuberculosis strains exhibiting resistance to less than or up to three anti-tubercular drugs, use of less than or up to three second line drugs in treatment and no change of regimen during treatment. CONCLUSION: Default from treatment was observed to be a major challenge in the treatment of MDR-TB due to long duration and expense of ATT.

Knowledge, attitude and practices regarding tuberculosis and dots among interns in delhi, India.

OBJECTIVE: To assess the knowledge, attitude and practices regarding tuberculosis (TB) and DOTS among young medical graduates (interns). DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Lady Hardinge Medical College and Maulana Azad Medical College, New Delhi, India, during November and December 2002, simultaneously. SUBJECTS AND METHODS: A pre-tested semi-structured questionnaire consisting of 30 items was administered to the young medical graduates/interns posted in different departments. The questions covered mode of transmission, symptoms of pulmonary TB, investigations, short course chemotherapy including DOTS as well as conventional anti-tubercular treatment, special situations, health education and chemoprophylaxis. RESULTS: A total of 287 interns were surveyed. Majority of the study subjects (92.7%) ranked DOTS strategy as more successful for treating tuberculosis in comparison to self-administered therapy. However, a mere 4.2% study subjects were aware of all modes of transmission. One hundred and eighty-nine (65.9%) correctly chose sputum examination for acid fast bacilli as the single most confirmatory test for diagnosing pulmonary TB. The rest either gave incorrect responses (including ELISA-17.0%, PCR-9.8%, X-ray chest-4.2%, ESR-1.7%) or did not respond (1.4%). Only 2.1% marked pyrazinamide and rifampicin as the agents to be avoided in patients with liver disease. One hundred and forty-one different treatment regimens were mentioned in the responses received and of those only 11(7.8%) were scientifically acceptable. CONCLUSION: TB is a major health problem in South-East Asian countries. There is a need for appropriate changes to be made in the undergraduate medical teaching/training curriculum in the concerned countries with regard to TB.

Green tea polyphenols prevent ultraviolet light-induced oxidative damage and matrix metalloproteinases expression in mouse skin.

Chronic exposure of solar ultraviolet (UV) light to human skin results in photoaging. UV-induced oxidative damage and induction of matrix metalloproteinases (MMP) have been implicated in this process. Because polyphenols from green tea (GTP) prevent other cutaneous adverse effects of UV radiation we hypothesized that UV irradiation-induced oxidative damage and induction of MMP might be prevented in vivo in mouse skin by oral administration of GTP. GTP was administered in drinking water (0.2%, wt/vol) to SKH-1 hairless mice, which were then exposed to multiple doses of UVB (90 mJ per cm2, for 2 mo on alternate days) following in vivo photoaging animal protocol. Treatment of GTP resulted in inhibition of UVB-induced protein oxidation in vivo in mouse skin, a hallmark of photoaging, when analyzed biochemically, by immunoblotting, and immunohistochemistry. GTP treatment also inhibited UVB-induced protein oxidation in vitro in human skin fibroblast HS68 cells, which supports in vivo observations. Moreover, oral administration of GTP also resulted in inhibition of UVB-induced expression of matrix degrading MMP, such as MMP-2 (67%), MMP-3 (63%), MMP-7 (62%), and MMP-9 (60%) in hairless mouse skin. These data suggest that GTP as a dietary supplement could be useful to attenuate solar UVB light-induced premature skin aging.

EGCG down-regulates telomerase in human breast carcinoma MCF-7 cells, leading to suppression of cell viability and induction of apoptosis.

Telomerase is elevated in >90% of breast carcinomas and therefore has received much attention as a target for breast cancer therapy and cancer diagnostic research. Dietary components that are capable of inhibiting the growth of cancer cells without affecting the growth of normal cells are receiving considerable attention in developing novel cancer-preventive approaches. Studies have shown that (-)-epigallocatechin-3-gallate (EGCG) from green tea imparts a growth inhibitory effect on cancer cells. Here, we show that treatment of EGCG dose-dependently inhibited (20-100%) the reproductive or colony forming potential, and also decreased cell viability at different time points studied ( approximately 80% inhibition) in human breast carcinoma MCF-7 cells but had no adverse effect on the growth of normal mammary cells. Treatment of EGCG for 48 and 72 h markedly increased the percentage of apoptotic cells (32-51%) in MCF-7 cells compared to that of non-EGCG treated cells (8-14%). In order to identify the possible mechanism of decreased cell viability and induction of apoptosis in breast carcinoma cells by EGCG, we found that treatment of MCF-7 cells with EGCG dose-dependently inhibited telomerase activity (40-55%), and also inhibited the mRNA expression (40-55%) of hTERT, a catalytic subunit of telomerase. Additional studies demonstrated that EGCG also inhibited the protein expression of hTERT, which indicated that inhibition of telomerase was associated with down-regulation of hTERT. Together, our results indicate that EGCG down-regulates telomerase in human breast carcinoma MCF-7 cells, leading to the suppression of cell viability and induction of apoptosis, thus providing the molecular basis for the development of EGCG as a novel chemopreventive and pharmacologically safe agent against breast cancer.

Proanthocyanidins from grape seeds inhibit expression of matrix metalloproteinases in human prostate carcinoma cells, which is associated with the inhibition of activation of MAPK and NF kappa B.

Prostate cancer (PCA) is the second most frequently diagnosed and leading cause of cancer-related deaths in men in the USA. The recognition that matrix metalloproteinases (MMPs) facilitate tumor cell invasion and metastasis of PCA has led to the development of MMP inhibitors as cancer therapeutic agents. As part of our efforts to develop newer and effective chemopreventive agents for PCA, we evaluated the effect of proanthocyanidins from grape seeds (GSP) on metastasis-specific MMP-2 and -9 in human prostate carcinoma DU145 cells by employing western blot and gelatinolytic zymography. Treatment of GSP dose-dependently inhibited cell proliferation (15-100% by 5-80 microg/ml of GSP), viability (30-80% by 20-80 microg/ml of GSP) and fibroblast conditioned medium (FCM)-induced expression of MMP-2 and -9 in DU145 cells. Since the signaling cascade of mitogen-activated protein kinases (MAPK) have been shown to regulate the expression of MMPs in tumor cells, we found that the treatment of DU145 cells with GSP (20-80 microg/ml) resulted in marked inhibition of FCM-induced phosphorylation of extracellular signal regulated kinase (ERK)1/2 and p38 but had little effect on c-Jun N-terminal kinase under similar experimental conditions. GSP treatment (20-80 microg/ml) to DU145 cells also dose-dependently inhibited FCM-induced activation of NF kappa B concomitantly with inhibition of MMP-2 and -9 expression in the same system. Additionally, the treatment of inhibitors of MEK (PD98059) and p38 (SB203580) to DU145 cells resulted in the reduction of FCM-induced phosphorylation of ERK1/2 and p38 concomitantly marked reduction in MMP-2 and -9 expressions. In further studies, treatment of androgen-sensitive LNCaP cells with a synthetic androgen R1881, resulted in an increase of MMP-2 and -9, which were completely abrogated in the presence of GSP (20-60 microg/ml). These data suggest that inhibition of metastasis-specific MMPs in tumor cells by GSP is associated with the inhibition of activation of MAPK and NF kappa B pathways, and thus provides the molecular basis for the development of GSP as a novel chemopreventive agent for both androgen-sensitive and -insensitive prostate cancer therapies.

Dietary feeding of proanthocyanidins from grape seeds prevents photocarcinogenesis in SKH-1 hairless mice: relationship to decreased fat and lipid peroxidation.

The use of dietary botanicals is receiving considerable interest in the protection of skin from the adverse biological effects of solar ultraviolet (UV) radiation. Dietary feeding of proanthocyanidins extracted from grape seeds (GSP) (0.2 and 0.5%, w/w) in AIN76 control diet to SKH-1 hairless mice resulted in prevention of photocarcinogenesis in terms of tumor incidence (20-95%), tumor multiplicity (46-95%) and tumor size (29-94%) against UVB-induced complete (both initiation + promotion), initiation and promotion stages of photocarcinogenesis. Feeding of GSP (0.5%, w/w) also resulted in prevention of malignant transformation of UVB-induced papillomas to carcinomas in terms of carcinoma incidence (45%), carcinoma multiplicity (61%) and carcinoma size (75%) compared with non-GSP treated mice following UVB-induced complete carcinogenesis protocol at the end of 30 weeks. Biochemical analysis revealed that treatment of GSP in vivo and in vitro systems significantly inhibited UVB- or Fe3+-induced lipid peroxidation by 57-66% (P<0.01) and 41-77% (P< 0.05-0.001), respectively, thus suggesting the antioxidant mechanism of photoprotection by GSP. Long-term feeding of GSP did not show apparent signs of toxicity in mice when determined in terms of body weight, diet consumption and physical characteristics of internal body organs like spleen, liver and kidney. Feeding of GSP also did not show apparent signs of toxicity when determined in terms of total body mass (mass of lean + fat), total bone mineral density and total bone mineral content by employing dual-energy X-ray absorptiometry (DXA). DXA analysis also revealed that feeding of GSP significantly decreased tissue fat level (24-27%, P<0.05) without changing the total body mass of the animals compared with non-GSP-fed animals. This can be attributed to increased lipolysis or decreased synthesis of fat due to administration of GSP. Together, it can be suggested that inhibition of photocarcinogenesis by GSP treatment may be associated with the reduction in UVB-induced oxidative damage and tissue fat content.

CD11b+ cells are the major source of oxidative stress in UV radiation-irradiated skin: possible role in photoaging and photocarcinogenesis.

Exposure of skin to solar UV radiation induces oxidative stress and suppression of cell-mediated immune responses. These effects are associated with the greater risk of several skin disorders including photoaging and photocarcinogenesis. We have shown that UV-induced infiltrating leukocytes contribute in developing oxidative stress in UV-irradiated skin. The peak period of UV-induced infiltrating leukocytes lies between 48 and 72 h after UV exposure of the skin. In this study we demonstrated that UV (90 mJ/cm2)-induced infiltrating CD11b+ cells in C3H/HeN mice skin were the major source of oxidative stress. Hydrogen peroxide (H2O2) was determined as a marker of oxidative stress. Flow cytometric analysis of viable cells revealed that the number of CD11b+H2O2+ cells were significantly higher (31.8%, P < 0.001) in UV-irradiated skin in comparison with non-UV-exposed skin (0.4%). Intraperitoneal administration of monoclonal antibodies to CD11b (rat IgG2b) to C3H/HeN mice inhibited UVB-induced infiltration of leukocytes, as evidenced by reduction in myeloperoxidase activity (64-80%, P < 0.0005), concomitant with significant reduction in H2O2 production both in epidermis and dermis (66-83%, P < 0.001-0.0005) when compared with the administration of rat IgG2b isotype of anti-CD11b. Furthermore, CD11b+ and CD11b- cell subsets were separated by immunomagnetic cell isolation technique from total epidermal and dermal single cell suspensions obtained 48 h after UV irradiation of the skin and analyzed for H2O2 production. Analytical data revealed that CD11b+ cell population from UV-irradiated skin resulted in significantly higher production of total H2O2 in both epidermis and dermis (87-89%, P < 0.0001) in comparison with CD11b- cell population (11-13% of total H2O2). These data revealed that infiltrating CD11b+ cells were the major source of oxidative stress in UV-irradiated skin and thus may contribute to photoaging and promotion of skin tumor growth within the UV-irradiated skin. Together, these data suggest that reduction in UV-induced skin infiltration of CD11b+ cells may be an alternative and effective strategy to reduce solar UV light-induced oxidative stress-mediated skin disorders including photoaging and photocarcinogenesis.