Reproductive Biology of Rheum webbianum Royle, a Vulnerable Medicinal Herb From Alpines of North-Western Himalaya.

Information on reproductive biology and pollination ecology studies of threatened plants are essential to develop strategies for their sustainable utilization and effective conservation. As such, these studies were conducted on Rheum webbianum, a high-value "vulnerable" medicinal herb of the north-western Himalaya. This species presents a unique mode of reproductive behavior through the involvement of different floral events, including the movement of reproductive organs. The plants survive extremely cold conditions through underground perennating rhizomes that sprout into juvenile shoots with the onset of the favorable climatic conditions. The peduncle arises from the axils of the radical leaves, bearing a globular collection of densely arranged hermaphrodite flowers with temporally separated male and female phases; the male phase precedes the female phase (protandry). Anther dehiscence and stigma receptivity is post-anthesis. Anthers dehisce longitudinally along margins, liberating a large mass of spherical and tricolpate pollen with spinulose exine. Pollen viability decreased to < 10% on day 9. Pistil is tristylous, with each style terminating into a fan-shaped stigma lobe. The pollen receptive surface of each stigmatic lobe remains incurved at an angle of 360° and shows upward movement after anthesis, forming a funnel-like structure at an angle of 180° with respect to the ovary. Pollination syndrome is ambophilous. Spontaneous autogamy or geitonogamy to a certain extent is achieved in this species due to the arrangement of flowers in the inflorescence and overlapping of male and female reproductive phases among them. Incurved stigmatic lobes and outward movement of stamens too facilitate outcrossing. Pollen/ovule ratio estimates, results of pollination experiments on breeding behavior, outcrossing, and self-compatibility indices demonstrated that plants are self-compatible and cross-fertile.

Expression of MiR-335 and its target metalloproteinase genes: clinical significance in breast cancer.

BACKGROUND: Early diagnosis of breast cancer decreases mortality rate; therefore, novel diagnostic methods are urgently required. In this study, authors aimed to investigate the role of serum-derived miR-335 in breast cancer, and the expression of matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9) and evaluating their feasibility and clinical utility as biomarkers for the early detection of breast cancer. MATERIALS AND METHODS: Blood samples were collected from a total of 210 individuals who were enrolled in this study. The participants were divided into newly diagnosed breast cancer patients (n = 115), patients with benign breast lesions (n =55) and healthy individuals as control group (n =40). The expression profile of miR-335, MMP2 and MMP9 were determined using quantitative polymerase chain reaction (qPCR). RESULTS: MiR 335 expression level was down-regulated in primary breast cancer group as compared to benign breast group and healthy individuals with 98% and 94.9% sensitivity and specificity, respectively. MMP2 and MMP9 showed significantly higher expression levels in breast cancer group as compared to both benign and healthy group and reporting 92.7% and 93% sensitivity, respectively. The relations between investigated markers and pathologic types, staging, grading, and lymph node involvement were significant with these factors. Expression level of miR-335 was decreased with increased MMP2 and MMP9 at significant level. CONCLUSION: MiR-335, MMP2, and MMP9 can be used as diagnostic markers in breast cancer.

Development of efficient strain of Ganoderma lucidum for biological stripping of cotton fabric dyed Reactive Blue 21.

One of the most common dyeing problems of textile industries is uneven and faulty dyeing over the finished quality of fabrics due to different reasons. These problems are usually tackled through chemical degradation in which uneven and faulty dye is removed from the surface of fiber but fabric quality is compromised. Chemical process also reduces the strength of the fabric and durability of textile material by reduction in reactive dye ability. The fabric cannot be reused due to the reduced strength. To overcome above mentioned problem, biological method of stripping in which enzymes produced by different micro-organisms are used. This process has no harmful effect on the fabric and is safe for environment. In this research work reactive blue 21 dye with 0.5, 2 and 4% shade strengths was used to dye cotton fabric. The Ganoderma lucidum fungal strains were mutated by UV mutagen, and five were selected for further processing. These mutant strains were grown at temperature ranges (20 °C to 40 °C); pH(3-5); inoculum size(1-5 mL) and fermentation time (3-15 days) . The required nutrients media to produce the ligninolytic enzymes was added to the flask. The strain which gave the fast decolourization results was selected for further optimization. Optimization was done by observing the variables: incubation time 12 days, pH 4, temperature 30 °C, and inoculum size 3 mL by applying Response Surface Methodology (RSM) in Central Composite Design (CCD). During the process of fabric color stripping, the enzyme assay revealed that the respective mutant UV-60 strain produced active enzymes with their Vmax, Mnp (427U/mL), LiP (785U/mL), and Lac (75 U/mL) enzymes decolorized 89% of the dye which is 25% more than the parent strain and also the production of enzyme is Mnp (344U/mL), LiP (693U/mL), and Lac (59 U/mL) enzymes which is lower than mutant strain.

Deciphering allelic variability and population structure in buckwheat: An analogy between the efficiency of ISSR and SSR markers.

Food and nutritional security continue to be the issues of concern in developing countries like ours. Exploring the reservoir of high potential unexplored genetic resources could address the world's food and nutritional insecurity. The availability of diverse data and the population structure of any crop germplasm is a valuable genetic resource for discovering genes that can help achieve food and nutritional stability. We used seven ISSR and seven SSR markers to investigate diversity among 63 buckwheat genotypes, including landraces from India's northwestern Himalayas. Various parameters such as percent polymorphism, PIC, resolving power, and marker index was used to evaluate the inequitable efficacy of these markers. We foundthat both marker systems are effective in detecting polymorphism in buckwheat germplasm. Seven ISSRs produced 55 polymorphic bands, while seven SSRs produced 32bands. When compared to ISSRs, SSRs had a greater average PIC value (0.43) than that of (0.36). ISSRs, on the other hand, had a resolving power of (4.38) compared to (1.42) for SSRs. The hierarchical cluster analysis dendrogram divided genotypes into three major clusters. We found that both marker systems were equally accurate in grouping buckwheat genotypes according to their geographical origins. Using 7 ISSR and 7 SSR markers, the model-based STRUCTURE analysis established a population with two sub-populations that correspond to species-based groupings. Within the population, there was a high level of genetic diversity. These results have consequences for both buckwheat breeding and conservation efforts.

Antagonistic, Anti-oxidant, Anti-inflammatory and Anti-diabetic Probiotic Potential of Lactobacillus agilis Isolated From the Rhizosphere of the Medicinal Plants.

Potential probiotic bacteria can be used as a biotherapeutic agent and a sustainable alternative to antibiotics, as an anti-oxidative, anti-inflammatory, and anti-diabetic agent without causing any serious side effects. Mostly human-friendly Lactic acid bacteria (LAB) have been isolated from the animal-human origin to be used as biotherapeutic agents or to produce useful metabolites (nutraceutical). However, less information is known about the role of medicinal plants associated LAB as biotherapeutic agents. The isolation of 115 human-friendly Lactobacillus strains was done from the rhizosphere of the medicinal plants Ocimum tenuiflorum, Azadirachta indica, Ficus carica. The obtained bacteria were then tested for their safe status before being using it for a beneficial purpose. Out of 115 strains, 29 (25%) were negative for blood hemolytic activities. Among these 29 isolates, three isolates did not show a breakdown of gelatin and were recognized as safe. Antibiotic resistance assay showed resistance of two of them against antibiotics discs of Streptomycin (10 µg), Ciprofloxacin (20 µg), Vancomycin (30 µg), Metronidazole (10 µg), Ampicillin (5 µg), Chloramphenicol (30 µg), Kanamycin (30 µg), Erythromycin (15 µg), Penicillin (10 µg) and Tetracycline (30 µg). The bacterial isolate (T-2) was found safe that was identified as Lactobacillus agilis by sequence analysis of 16 s rRNA gene and processed in vitro as an anti-bacterial, anti-oxidant, anti-diabetic, and anti-inflammatory agent. Free radical scavenging activities and inhibition of α-amylase activities for Lactobacillus agilis were found relative to standard drug values as 68% and 73% and 51.3% and 65.3%, respectively. The in-vitro anti-inflammatory assay showed 61.6% (Lactobacillus agilis) while showed 69% (aspirin) activity for denaturation albumin protein. The results suggested that Lactobacillus agilis can be used as a potential probiotic strain as well as can be used to produce nutraceuticals.

Ecological assessment and environmental niche modelling of Himalayan rhubarb (Rheum webbianum Royle) in northwest Himalaya.

In an era of anthropocene, threatened and endemic species with small population sizes and habitat specialists experience a greater global conservation concern in view of being at higher risk of extinction. Predicting and plotting appropriate potential habitats for such species is a rational method for monitoring and restoring their dwindling populations in expected territories. Ecological niche modelling (ENM) coalesces species existence sites with environmental raster layers to construct models that describe possible distributions of plant species. The present study is aimed to study the potential distribution and cultivation hotspots for reintroducing the high value, vulnerable medicinal herb (Rheum webbianum) in the Union territories of Jammu and Kashmir and Ladakh using population attributes and ecological niche modelling approach. Sixty-three populations inventoried from twenty-eight areas display a significant change in the phytosociological attributes on account of various anthropogenic threats. The current potential habitats coincide with actual distribution records and the mean value of Area Under Curve (AUC) was 0.98 and the line of predicted omission was almost adjacent to omission in training samples, thus validating a robustness of the model. The potential habitat suitability map based on the current climatic conditions predicted a total of 103760 km2 as suitable area for the growth of Rheum webbianum. Under the future climatic conditions, there is a significant reduction in the habitat suitability ranging from -78531.34 Km2 (RCP 4.5 for 2050) to -77325.81 (RCP 8.5 for 2070). Furthermore, there is a slight increase in the suitable habitats under future climatic conditions, ranging from +21.99 Km2 under RCP 8.5 (2050) to +3.14 Km2 under RCP 4.5 (2070). The Jackknife tests indicated Precipitation of Driest Month (BIO14) as the most contributing climatic variable in governing the distribution of R. webbianum. Therefore, scientifically sound management strategies are urgently needed to save whatever populations are left in-situ to protect this species from getting extinct. Present results can be used by conservationists for mitigating the biodiversity decline and exploring undocumented populations of R. webbianum on one hand and by policymakers in implementing the policy of conservation of species with specific habitat requirements by launching species recovery programmes in future on the other.

Genome-Wide Transcriptome Profiling, Characterization, and Functional Identification of NAC Transcription Factors in Sorghum under Salt Stress.

Salinity stress has become a significant concern to global food security. Revealing the mechanisms that enable plants to survive under salinity has immense significance. Sorghum has increasingly attracted researchers interested in understanding the survival and adaptation strategies to high salinity. However, systematic analysis of the DEGs (differentially expressed genes) and their relative expression has not been reported in sorghum under salt stress. The de novo transcriptomic analysis of sorghum under different salinity levels from 60 to 120 mM NaCl was generated using Illumina HiSeq. Approximately 323.49 million high-quality reads, with an average contig length of 1145 bp, were assembled de novo. On average, 62% of unigenes were functionally annotated to known proteins. These DEGs were mainly involved in several important metabolic processes, such as carbohydrate and lipid metabolism, cell wall biogenesis, photosynthesis, and hormone signaling. SSG 59-3 alleviated the adverse effects of salinity by suppressing oxidative stress (H2O2) and stimulating enzymatic and non-enzymatic antioxidant activities (SOD, APX, CAT, APX, POX, GR, GSH, ASC, proline, and GB), as well as protecting cell membrane integrity (MDA and electrolyte leakage). Significant up-regulation of transcripts encoding the NAC, MYB, and WRYK families, NHX transporters, the aquaporin protein family, photosynthetic genes, antioxidants, and compatible osmolyte proteins were observed. The tolerant line (SSG 59-3) engaged highly efficient machinery in response to elevated salinity, especially during the transport and influx of K+ ions, signal transduction, and osmotic homeostasis. Our data provide insights into the evolution of the NAC TFs gene family and further support the hypothesis that these genes are essential for plant responses to salinity. The findings may provide a molecular foundation for further exploring the potential functions of NAC TFs in developing salt-resistant sorghum lines.

Aberrant Lighting Causes Anxiety-like Behavior in Mice but Curcumin Ameliorates the Symptoms.

In the modern research field, laboratory animals are constantly kept under artificial lighting conditions. However, recent studies have shown the effect of artificial light on animal behavior and metabolism. In the present study on mice, following three weeks of housing in dim light at night (dLAN; 5lux) and complete darkness (DD; 0lux), we monitored the effect on body weight, daily food intake, anxiety-like behavior by employing the open field test, and expression of the period (PER1) gene. We also studied the effect of oral administration of different concentrations of curcumin (50, 100, and 150 mg/kg) for three weeks in the same mice and monitored these parameters. The exposure to dLAN had significantly increased the anxiety-like behavior and body weight possibly through the altered metabolism in mice, whereas exposure to DD caused increased anxiety but no significant difference in weight gain. Moreover, the expression of the PER1 gene involved in sleep was also found to be decreased in the aberrant light conditions (dLAN and DD). Although the treatment of curcumin had no effect on body weight, it ameliorated the anxiety-like behavior possibly by modulating the expression of the PER1 gene. Thus, alteration in the light/dark cycle had a negative effect on laboratory animals on the body weight and emotions of animals. The present study identifies the risk factors associated with artificial lighting systems on the behavior of laboratory animals and the ameliorative effects of curcumin, with a focus on anxiety-like behavior.

Clinical impact of PTEN methylation status as a prognostic marker for breast cancer.

BACKGROUND: Aberrant DNA methylation of phosphatase and tensin homolog (PTEN) gene has been found in many cancers. The object of this study was to evaluate the clinical impact of PTEN methylation as a prognostic marker in breast cancer. The study includes 153 newly diagnosed females, and they were divided according to their clinical diagnosis into breast cancer patients (n = 112) and females with benign breast lesion (n = 41). A group of healthy individuals (n = 25) were recruited as control individuals. Breast cancer patients were categorized into early stage (0-I, n = 48) and late stage (II-III, n = 64), and graded into low grade (I-II, n = 42) and high grade (III, n = 70). Their pathological types were invasive duct carcinoma (IDC) (n = 66) and duct carcinoma in situ (DCI) (n = 46). Tumor markers (CEA and CA15.3) were detected using ELISA. DNA was taken away from the blood, and the PTEN promoter methylation level was evaluated using the EpiTect Methyl II PCR method. RESULTS: The findings revealed the superiority of PTEN methylation status as a good discriminator of the cancer group from the other two groups (benign and control) with its highest AUC and increased sensitivity (96.4%) and specificity (100%) over tumor markers (50% and 84% for CEA and 49.1% and 86.4% for CA15.3), respectively. The frequency of PTEN methylation was 96.4% of breast cancer patients and none of the benign and controls showed PTEN methylation and the means of PTEN methylation (87 ± 0.6) were significantly increased in blood samples of breast cancer group as compared to both benign and control groups (25 ± 0.7 and 12.6 ± 0.3), respectively. Methylation levels of PTEN were higher in the blood of patients with ER-positive than in patients with ER-negative cancers (P = 0.007) and in HER2 positive vs. HER2 negative tumors (P = 0.001). The Kaplan-Meier analysis recognizes PTEN methylation status as a significant forecaster of bad progression-free survival (PFS) and overall survival (OS), after 40 months follow-up. CONCLUSIONS: PETN methylation could be supposed as one of the epigenetic aspects influencing the breast cancer prognosis that might foretell more aggressive actions and worse results in breast cancer patients.

Mini-mental state examination (MMSE) scores in elderly Jordanian population.

Background: The Mini-Mental State Examination (MMSE) is a simple informative and validated screening test of cognitive functions. No data of MMSE scores has been published about elderly Jordanian population. Objective: To study the MMSE scores in the elderly Jordanian population (defined as age of 50 years or more). Methods: This was a cross sectional study. A 250 healthy elderly Jordanians were interviewed and tested for their MMSE score. Their scores were analyzed and the effect of age, gender, education, marital status, diabetes mellitus (DM), hypertension (HTN), smoking, dyslipidemia, heart disease (HD), and family history of dementia (FHD) on the score was studied. Results: MMSE scores of 236 elderly subjects, aged 63±8.4 years (range 50-86 years) were analyzed. There were 111 (47%) males. There was a direct correlation between MMSE score and education level. People with a higher education (college) had the highest scores in comparison with people who are illiterates or have a high school level. The score correlated negatively with age (Pearson correlation r = -0.23, p = 0.00). Other studied variables did not correlate with score on multivariate analysis. Conclusion: Jordanians ≥50 years old with no previous brain disease scored 26.7 ± 3.2. Education was the most important determining factor of this score. the score showed also a negative correlation with age. We suggest that in this population a cut off score of 24 for those with high education beyond high school and 21 for those with up to high school education and it is not appropriate for illiterate.

Clinical aspects of circulating miRNA-335 in breast cancer patients: A prospective study.

BACKGROUND: The impact of aberrant expression of miRNAs, small noncoding RNAs of 19 to 23 nucleotide, has been reported in different types of cancer, such as breast cancer. Authors aim to investigate the role of circulating miRNA-335 as a diagnostic and prognostic marker for breast cancer. MATERIALS AND METHODS: miRNA-335 expression was measured in primary breast cancer patients (n = 106), patients with benign breast lesion (n = 49) and healthy individuals as control (n = 40) using quantitative real-time polymerase chain reaction and its diagnostic efficacy, relation with clinicopathological factors, and disease-free survival (DFS) and overall survival (OS) were assessed. RESULTS: A significant decrease in miRNA-335 expression was reported in patients with breast cancer as compared to the other two investigated groups. The positivity rate for miRNA were related to adverse clinical features of primary breast cancer as high histological grading (X2 = 7.72, P = 0.016), presence of metastasis to lymph node (X2 = 21.8, P < 0.001), large tumor size (X2 = 6.41, P = 0.041), and hormonal status (P < 0.001). miRNA-335 mean rank level was significantly different among breast cancer subtypes and its level was inferior in triple negative breast cancer. Worse DFS (X2 = 7.76, P = 0.005) and OS (X2 = 9.3, P = 0.002) were reported with decreased miRNA-335 level. CONCLUSION: Assessment of circulating miRNA expression level is a promising minimal invasive marker for diagnosis and prediction of breast cancer prognosis with significant discrepancies among molecular breast cancer subtypes.

Effectiveness of vinpocetine and isosorbide-5-mononitrate on experimental schistosomiasis mansoni: Biochemical and immunohistochemical study.

Schistosomiasis is one of the most important tropical and subtropical devastating diseases, where praziquantel is the sole drug of choice. Praziquantel effectively kills the adult worms, however, drug resistance has been repeatedly reported. Moreover, there is currently no efficient anti-fibrotic therapy available for chronic schistosomiasis. So, novel drugs which exert anti-fibrotic efficacy are urgently needed. This research is complementary to our previous work that evaluated the anti-schistosomal effects of the anti-inflammatory vinpocetine, as well as the vasodilator and the anti-oxidant isosorbide-5-mononitrate. In the present study, we assessed the therapeutic efficacies of drugs in Swiss albino female mice experimentally infected with an Egyptian strain of Schistosoma mansoni, using some biochemical and immunohistochemical parameters. Our results revealed that both vinpocetine and isosorbide-5-mononitrate monotherapy significantly decreased hepatic nuclear factor-kappaB, 10 weeks post infection. The best effects were seen in mice administered praziquantel combined with isosorbide-5-mononitrate, as detected by reduction in hydroxyproline and collagen contents of the liver, and significant increase in the hepatic nitric oxide content. The data provides insight into the potential effects of the assessed drugs with isosorbide-5-mononitrate being more superior to vinpocetine, hence it can be used as novel adjuvant to praziquantel to alleviate schistosomal hepatic fibrosis. However, molecular mechanism/s and clinical trials are worthy to be scrutinized.

Aberrant Expression of Some Circulating miRNAs in Childhood Acute Lymphoblastic Leukemia.

Acute lymphoblastic leukemia (ALL) is a heterogeneous cancer commonly affecting children due to dysregulation of miRNA expression. In the current study, authors investigated the expression profile for miRNA-125b-1 and miRNA-203 among childhood ALL. Blood samples were collected from newly diagnosed childhood ALL and healthy control children. The expression profile for candidate miRNAs was detected using quantitative RT-PCR analysis. Statistical analysis were performed using receiver operating characteristic curve (ROC) to examine the diagnostic efficacy of the two miRNA and their levels among ALL clinicopathological factors and phenotypes. The median expression level for miRNA-125b-1 was significantly high in childhood ALL; while miRNA-203 level was significantly low in childhood ALL as compared to control ones. MiRNA-125-1 reported significant increase in T-ALL as compared to other ALL phenotypes. Median miRNA-203 level was high in T-ALL followed by pre-B-ALL although no significant difference was reported. Clinicopathological factors did not emphasize significance with either detected miRNAs. Using ROC curve the diagnostic efficacy was significant with an area under the curve 0.858 for miRNA-125b-1 (83.72, 100%) and 0.878 for miRNA-203 (97.67, 86.96%). The combination of the two key miRNAs revealed absolute sensitivity (100%). MiRNA-125b-1 and miRNA-203 can be useful molecular markers for diagnosis of ALL. Further studies with large cohort are warranted to validate these results.

A comparative study on the anti-schistosomal and hepatoprotective effects of vinpocetine and isosorbide-5-mononitrate on Schistosoma mansoni-infected mice.

Schistosomiasis is a remarkable public health problem in developing countries. Presently, praziquantel is the optional drug for all human schistosomiasis. Owing to the increased praziquantel resistance, there is an urgent need to develop new alternatives. This study aims at determining the anti-schistosomal and/or the hepatoprotective effects of the anti-inflammatory drug; vinpocetine, and the vasodilator and the nitric oxide donor; isosorbide-5-mononitrate, in comparison to praziquantel. In the present research, the therapeutic efficacies of these drugs were assessed in Swiss albino female mice (CD-I strain) experimentally infected with an Egyptian strain of Schistosoma mansoni, using some general, parasitological, and histopathological parameters. In this work, praziquantel significantly reduced worm burden and hepatic egg load, increased the percentage of dead eggs in the small intestine and decreased granuloma count, but did not reduce granuloma diameter. While, either vinpocetine or isosorbide-5-mononitrate monotherapy did not induce significant reduction in the worm count, hepatic egg load or shift in the oogram pattern, but significantly reduced granuloma count and diameter. Moreover, isosorbide-5-mononitrate significantly reduced hepatic inflammation and necrosis. The best results were obtained in the mice groups treated with isosorbide-5-mononitrate combined with praziquantel or vinpocetine. Our results point to vinpocetine and isosorbide-5-mononitrate as a convenient and promising adjuvant to praziquantel for ameliorating schistosomal liver pathology. Further studies are recommended to reveal the actual pathways responsible for the different activities of vinpocetine and isosorbide-5-mononitrate.

Diagnostic role of glypican 3 and CD34 for differentiating hepatocellular carcinoma from nonmalignant hepatocellular lesions.

Well-differentiated hepatocellular carcinoma (HCC) may be difficult to distinguish from a benign lesion. Glypican 3 (GPC-3) is an oncofetal protein, which has been demonstrated to be up-regulated in HCC. The aim of this study is to evaluate the diagnostic role of combined GPC-3 and CD34 immunoassaying in the distinction between HCC and benign hepatic mimickers. This study was performed on 100 cases of formalin-fixed, paraffin-embedded cases of hepatic focal lesions obtained from the files of pathology laboratory of our university from 2009 to 2012. The following groups were studied: group A (n = 60) (hepatocellular malignant lesions) and group B (n = 40) (Hepatocellular nonmalignant lesions). All cases were stained with GPC-3 and CD34. Sensitivity, specificity, and positive and negative predictive values were calculated for both antibodies. Glypican 3 and complete CD34 staining pattern expression in group A was significantly higher than in group B. The results of costaining showed that, in HCCs, almost all the GPC-3-positive cases had a complete CD34 staining pattern, whereas in the 40 hepatocellular nonmalignant lesions, none stained up with the 2 markers. Therefore, although the sensitivity declined (82%), the specificity and positive predictive value (PPV) of costaining reached 100% and were greater than that observed for single staining with GPC-3 (specificity, 92.5%; PPV, 94.3%) or CD34 (specificity, 97.5%; PPV, 98.3%). Our data demonstrate that GPC-3 and CD34 costaining has better diagnostic value for differentiating HCC from nonmalignant hepatocellular lesions than does single staining.

Potential value of estrogen receptor beta expression in colorectal carcinoma: interaction with apoptotic index.

PURPOSE: The study was designed to investigate the clinicopathological correlations, relationship to apoptotic index, and prognostic significance of estrogen receptor beta expression in colorectal carcinoma. METHODS: The study was carried out on 40 patients with newly diagnosed colorectal cancer. The patients' data were collected prospectively and the 2 years overall survival was the endpoint. Estrogen receptor beta expression was assessed by immunohistochemistry. Apoptotic body index was calculated by counting apoptotic cells using the modified TUNEL assay. RESULTS: Estrogen receptor beta positivity was detected in 65% of colorectal cancer cases, while estrogen receptor alpha positivity was found in only 7% of cases. The rate of estrogen receptor beta immunoreactivity was significantly higher in low-grade colorectal tumors. The median apoptotic index in estrogen receptor beta positive cases was significantly higher than in estrogen receptor beta negative cases (6% versus 3%; p = 0.01). The median overall survival was higher in estrogen receptor beta positive cases (22 versus 18 months); however, the difference was not statistically significant. CONCLUSIONS: The study results reinforce the importance of the estrogen receptor beta rather than the estrogen receptor alpha in colorectal cancer. Lack of estrogen receptor beta expression is associated with loss of differentiation and decreased apoptosis. Future studies should include validation of estrogen receptor beta as a prognostic marker and exploration of its role as a target in the management of colorectal cancer.

Prognostic significance of telomerase activity and some tumor markers in non-small cell lung cancer.

The prognosis of non small cell lung cancer (NSCLC) has remained disappointing over the last decades even in localized stages. Numerous prognostic factors have been investigated which might select patients for additional treatment. The objective of the current study was to assess the prognostic significance of telomerase activity, serum anti-p53 antibodies (anti-p53a), c-erbB-2 and CEA in patients with NSCLC. The study included 60 patients with histological proven NSCLC besides 60 controls (30 smokers and 30 nonsmokers). Patients were divided into four stages according to their histopathology. All patients were subjected to; determination of telomerase activity by telomeric repeat amplification protocol (TRAP) assay in tumor tissue specimens and adjacent normal lung tissues, also, determination of preoperative serum anti-p53a, c-erbB-2 and CEA. Telomerase activity was detected in 40 of 60 (66.6%) of NSCLC tissue specimens using the TRAP assay. As regard the stages, telomerase activity was positive in 5 of 15 patients (33.3%) with stage I NSCLC, in 11 of 20 patients (55%) with stage II NSCLC, in 9 of 10 patients (90%) with stage III NSCLC and in all patients (100%) with stage IV NSCLC. More cases of positive telomerase activity were observed in the group with advanced disease and in the group with poorly differentiated tumors. Telomerase activity was not detected in any normal lung tissue. The concentrations of serum anti-p53a, c-erbB-2, CEA were significantly higher in patients with NSCLC in comparison to the smoker and nonsmoker controls and their levels increased according to the stage of disease. Logistic regression test showed a relation between telomerase positivity and anti- p53a but no relation with c-erbB2, CEA. Telomerase activity was detected in most of NSCLC tissues; it was detected more frequently in advanced disease than early-stage disease. Anti-p53, c-erbB-2 and CEA were significantly higher in patients with NSCLC than controls and this increment was more evident in late stages of the disease. So, these biological markers might be useful predictors of prognosis. They may be helpful in defining groups of patients with NSCLC who could benefit from adjuvant treatments, also these markers can be used as therapeutic targets.

Clinical significance of inflammatory markers in polycystic ovary syndrome: their relationship to insulin resistance and body mass index.

BACKGROUND: Women with polycystic ovary syndrome (PCOS) have an increased prevalence of insulin resistance (IR) and related disorders. Elevated serum levels of high sensitivity CRP (hs-CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) reflect low-grade chronic inflammation and have been associated with several insulin-resistant states; they are useful cardiovascular risk markers. The objective of this study was to investigate whether soluble inflammatory markers are altered in PCOS focusing on its relationship with obesity and indexes of insulin resistance. PATIENTS AND METHODS: One hundred and eight women with PCOS and 75 healthy women were recruited. Patients were divided according to body mass index (BMI) into two groups; group I (BMI < 27 kg/m(2)) and group II (BMI > or = 27 Kg/m(2)). Serum levels of hs-CRP, IL-6, and TNF-alpha, lipid and hormone profiles were measured. RESULTS: PCOS patients had increased levels of testosterone, luteinizing hormone (LH), androstendione, insulin level and HOMA index compared to healthy BMI matched controls. High-density lipoprotein (HDL) concentrations were significantly reduced in both patient groups compared to their controls, while triglyceride levels were significantly increased in obese group compared to controls. There were no significant difference in serum inflammatory markers hs-CRP, IL-6 and TNF-alpha between group I and their matched controls. On the other hand, there were significant increase in these markers between group II and their matched controls. There were highly significant positive correlation between hs-CRP and IL-6 (r=0.702, P< 0.001) and between hs-CRP and TNF-alpha (r=0.621, P<0.001), also between IL-6 and TNF-alpha (r=0.543, P< 0.001). These inflammatory markers correlated significantly with BMI and HOMA index. Multiple regression analysis revealed that BMI and HOMA were predictors of IL-6 levels (b=11.173, P< 0.001, b=13.564, P< 0.001 respectively) and BMI was the only predictor of hs-CRP levels (b=12.578, P< 0.001) and TNF-alpha levels (b=0.134, P<0.001). CONCLUSION: PCOS and obesity induce an increase in serum inflammatory cardiovascular risk markers. The precise mechanisms underlying these associations require additional studies to clarify the state of the cardiovascular system in women with PCOS compared with controls in large numbers of patients to determine the relative contribution of different factors including insulin resistance, androgen status and BMI.