Lordosis facilitated by GPER-1 receptor activation involves GnRH-1, progestin and estrogen receptors in estrogen-primed rats.

The present study assessed the participation of membrane G-protein coupled estrogen receptor 1 (GPER-1) and gonadotropin releasing hormone 1 (GnRH-1) receptor in the display of lordosis induced by intracerebroventricular (icv) administration of G1, a GPER-1 agonist, and by unesterified 17ß-estradiol (free E2). In addition, we assessed the participation of both estrogen and progestin receptors in the lordosis behavior induced by G1 in ovariectomized (OVX), E2-benzoate (EB)-primed rats. In Experiment 1, icv injection of G1 induced lordosis behavior at 120 and 240min. In Experiment 2, icv injection of the GPER-1 antagonist G15 significantly reduced lordosis behavior induced by either G1 or free E2. In addition, Antide, a GnRH-1 receptor antagonist, significantly depressed G1 facilitation of lordosis behavior in OVX, EB-primed rats. Similarly, icv injection of Antide blocked the stimulatory effect of E2 on lordosis behavior. In Experiment 3, systemic injection of either tamoxifen or RU486 significantly reduced lordosis behavior induced by icv administration of G1 in OVX, EB-primed rats. The results suggest that GnRH release activates both estrogen and progestin receptors and that this activation is important in the chain of events leading to the display of lordosis behavior in response to activation of GPER-1 in estrogen-primed rats.

Radio-induced apoptosis of peripheral blood CD8 T lymphocytes is a novel prognostic factor for survival in cervical carcinoma patients.

BACKGROUND AND PURPOSE: A close relationship exists between immune response and tumor behavior. This study aimed to explore the associations between radiation-induced apoptosis (RIA) in peripheral blood lymphocytes (PBL) and clinical pathological variables. Furthermore, it assessed the role of RIA as a prognostic factor for survival in cervical carcinoma patients. PATIENTS AND METHODS: Between February 1998 and October 2003, 58 consecutive patients with nonmetastatic, localized stage I-II cervical carcinoma who had been treated with radiotherapy (RT) ± chemotherapy were included in this study. Follow-up ended in January 2013. PBL subpopulations were isolated and irradiated with 0, 1, 2 and 8 Gy then incubated for 24, 48 and 72 h. Apoptosis was measured by flow cytometry and the ß value, a parameter defining RIA of lymphocytes, was calculated. RESULTS: Mean follow-up duration was 111.92 ± 40.31 months. Patients with lower CD8 T lymphocyte ß values were at a higher risk of local relapse: Exp(B) = 5.137, confidence interval (CI) 95 % = 1.044-25.268, p = 0.044. Similar results were observed for regional relapse: Exp(B) = 8.008, CI 95 % = 1.702-37.679, p = 0.008 and disease relapse: Exp(B) = 6.766, CI 95 % = 1.889-24.238, p = 0.003. In multivariate analysis, only the CD8 T lymphocyte ß values were found to be of prognostic significance for local disease-free survival (LDFS, p = 0.049), regional disease-free survival (RDFS, p = 0.002), metastasis-free survival (MFS, p = 0.042), disease-free survival (DFS, p = 0.001) and cause-specific survival (CSS p = 0.028). CONCLUSION: For the first time, RIA in CD8 T lymphocytes was demonstrated to be a predictive factor for survival in cervical carcinoma patients.

On-line surface plasmon resonance biosensing of vascular endothelial growth factor signaling in intact-human hepatoma cell lines.

Surface plasmon resonance (SPR) monitoring of biorecognition events at intracellular levels is a valuable tool for studying the angiogenic response of carcinoma living cells during tumor growth and proliferation. We report here a comparative study of two different strategies to detect human hepatoma cell interactions between transmembrane vascular endothelial growth factor receptor (VEGFR2) and vascular endothelial growth factor (VEGF). To monitor VEGFR2 activation after VEGF stimulation, intact hepatocellular carcinoma HepG2 or Huh7 cells (2 × 10(5) cells per mL) were directly immobilized on the sensor chip. Distinguishable SPR sensorgrams were obtained for each cell line depending on the time required for VEGFR2 activation. SPR signals for VEGF-VEGFR2 binding were inhibited by the VEGFR inhibitor, CBO-P11. The SPR response after VEGF stimulation/inhibition was in good agreement with the results observed by immunoblotting analysis. In a second approach we used intact cell lines as analytes. SPR analysis was done by injecting HepG2 and HuH7 cell suspensions (2-4 × 10(4) cells per mL) onto a sensor surface previously immobilized with VEGF via a thiol self-assembled monolayer (SAM). Specificity and reproducibility were evaluated reusing the same chip surface over more than 60 complete regeneration cycles. Comparison between both methods yielded differences in terms of reliability, making the latter strategy more effective for the analysis of real samples. The investigation of VEGF signaling in intact human hepatoma living cells by SPR monitoring comprises a novel and promising design for the study of tumor angiogenesis via downregulation of VEGF and VEGFR2 pathways. Further investigation on VEGFR activation and vascular function could contribute to establish a robust and meaningful tool for early cancer diagnostics.

Inhibition of VEGF expression through blockade of Hif1α and STAT3 signalling mediates the anti-angiogenic effect of melatonin in HepG2 liver cancer cells.

BACKGROUND: Hepatocellular carcinoma (HCC) growth relies on angiogenesis via vascular endothelial growth factor (VEGF) release. Hypoxia within tumour environment leads to intracellular stabilisation of hypoxia inducible factor 1 alpha (Hif1α) and signal transducer and activator of transcription (STAT3). Melatonin induces apoptosis in HCC, and shows anti-angiogenic features in several tumours. In this study, we used human HepG2 liver cancer cells as an in vitro model to investigate the anti-angiogenic effects of melatonin. METHODS: HepG2 cells were treated with melatonin under normoxic or CoCl2-induced hypoxia. Gene expression was analysed by RT-qPCR and western blot. Melatonin-induced anti-angiogenic activity was confirmed by in vivo human umbilical vein endothelial cells (HUVECs) tube formation assay. Secreted VEGF was measured by ELISA. Immunofluorescence was performed to analyse Hif1α cellular localisation. Physical interaction between Hif1α and its co-activators was analysed by immunoprecipitation and chromatin immunoprecipitation (ChIP). RESULTS: Melatonin at a pharmacological concentration (1 mM) decreases cellular and secreted VEGF levels, and prevents HUVECs tube formation under hypoxia, associated with a reduction in Hif1α protein expression, nuclear localisation, and transcriptional activity. While hypoxia increases phospho-STAT3, Hif1α, and CBP/p300 recruitment as a transcriptional complex within the VEGF promoter, melatonin 1 mM decreases their physical interaction. Melatonin and the selective STAT3 inhibitor Stattic show a synergic effect on Hif1α, STAT3, and VEGF expression. CONCLUSION: Melatonin exerts an anti-angiogenic activity in HepG2 cells by interfering with the transcriptional activation of VEGF, via Hif1α and STAT3. Our results provide evidence to consider this indole as a powerful anti-angiogenic agent for HCC treatment.

Water network cost optimization in a paper mill based on a new library of mathematical models.

The increasing costs associated with water supply and the disposal of wastewater has stimulated industries to seek more efficient water management systems. Mathematical modelling and simulation can be a very valuable tool for the study of the multiple alternatives available whilst assessing optimum solutions for water management in industry. This study introduces a new steady state model library able to reproduce industrial water circuits. It has been implemented in a novel software framework for the representation, simulation and optimization of industrial water networks. A water circuit representing a paper mill has been modelled and simulated showing the capability to reproduce real case studies. Alternative scenarios for the water network have also been tested to assess the capability of the models to optimize water circuits minimizing total cost.

Health related quality of life and late side effects of long-term survivors of Ewing's sarcoma of bone.

PURPOSE: The data examining the Health Related Quality of Life (HRQOL) after definitive treatment for Ewing's sarcoma (EWS) is sparse. The objective of this study was to assess the HRQOL and late side effects in EWS of bone survivors treated in the past 2 decades. METHODS: Seventeen long-term (≥ 5 years) EWS survivors (age range, 11-27 years) treated from 1990 to 2004 completed the EORTC-QLQ-C30. We compared the HRQOL of our data set with a reference group of cancer survivors and also with the general population. Musculoskeletal late treatment toxicity was also evaluated. The Mann-Whitney and Wilcoxon tests were used for analyses. RESULTS: EWS survivors of our series reported significantly better global, physical, role and cognitive HRQOL and less symptoms of fatigue, pain and insomnia than the normative sample of cancer survivors. Moreover, the HRQOL was equivalent to the general population, with the exception of social functioning. Additionally, patients with tumor location in the extremities did not report significant HRQOL differences compared with those with tumor at other locations. Common chronic grade 2 side effects were generalized muscle weakness (23%) and decreased joint range of motion (23%). Only one patient experienced musculoskeletal chronic grade 3 toxicity. CONCLUSION: Our findings suggest that EWS survivors treated in the modern era do not experience lower HRQOL than other cancer survivors. Rather, the HRQOL appears to be equivalent to the general population. Further and larger studies are needed to confirm these results.

Replacement of fresh water use by final effluent recovery in a highly optimized 100% recovered paper mill.

A further closure of the water circuit in paper mills with a relative high optimization of their water network is limited by the increase of contamination in the water and runnability problems of the paper machine. Therefore, new strategies for saving water must be focussed on the treatment of final effluents of the paper mill, aiming to obtain high quality water that may replace fresh water use in some applications. An appropriate treatment train performed at pilot scale, consisting on a previous clarification stage followed by anaerobic and aerobic treatments, ultrafiltration, and reverse osmosis, made possible producing the highest water quality from the final effluent of the mill. Anaerobic pre-treatment showed very good performance assisting the aerobic stage on removing organics and sulphates, besides it produced enough biogas for being considered as cost-effective. Permeate recovery depended on the silica content of the paper mill effluent, and it was limited to a 50-60%. The reject of the membranes fully met the legislation requirements imposed to effluents arriving to municipal wastewater treatment plants.

Two thalidomide analogs induce persistent estrous behavior and inhibit uterus contractility in rats: The central role of cAMP.

The effects of 4NO2PDPMe and 4APDPMe, which are thalidomide (Tha) analogs that act as selective phosphodiesterase (PDE-4) inhibitors, on estrous behavior (lordosis and proceptive behaviors) and on uterine contraction were studied in ovariectomized (OVX) estrogen-primed Sprague Dawley (SD) and in intact non-pregnant Wistar rats, respectively. We found that intracerebroventricular (ICV) infusion of either 4NO2PDPMe or 4APDPMe (20 to 80 µg) stimulated intense lordosis and proceptive behavior in response to mounts from a sexually active male, within the first 4 h after infusion, and persisting for up to 24 h. Inhibitors of the progesterone receptor (RU486, administered subcutaneously), the estrogen receptor (tamoxifen, ICV), the adenylate cyclase (AC)/ cyclic AMP (cAMP)/protein kinase A (PKA) pathway (administered ICV), and the mitogen activated protein kinase (MAPK) pathway (administered ICV) significantly decreased lordosis and proceptive behavior induced by Tha analogs. Uterine contractility studies showed that Tha analogs inhibited both the K+- and the Ca2+-induced tonic contractions in rat uterus. Tha analogs were equally effective, but 4APDPMe was more potent than 4NO2PDPMe. These results strongly suggest the central role of cAMP in both processes, sexual behavior, and uterine relaxation, and suggest that Tha analogs may also act as Ca2+-channel blockers.

Acid invertase localization in leaves of the fern Pteris deflexa link.

The localization of invertase, a key enzyme in plant carbohydrate metabolism, has been established in several higher plants, but there are no reports of it in ferns. The aim of the present work was to establish the localization of the previously reported acid invertase activity of Pteris deflexa in fronds tissues and to compare the findings with invertase localization in higher plants. Acid invertase, localized by immuno-histochemical and histochemical techniques on fresh tissues, was evident in vascular tissue, mainly in phloem. It was also detected in parenchymatic, sclerenchymatic and epidermic cells of petiole, rachis and rachis branches as well as in veins of leaf blades. Our results demonstrate that P. deflexa acid invertase localization is the same to that of higher plants. Hence, potential roles of the fern enzyme in relation to the storage and utilization of sucrose and to control carbon flux could be the same of those proposed to higher plants.

Radiotherapy in soft-tissue sarcoma of the extremities.

Sarcomas are an infrequent and heterogeneous group of neoplasia. Surgery with or without associated radiotherapy (RT) is the basic treatment for this type of tumour. To increase the therapeutic ratio (the index between cytotoxic effects in tumours and normal tissue complications with a certain dose of radiation), new advances are being investigated to increase local and distant control and to decrease the morbidity of the treatment. The aim of this review was to analyse the different strategies, based on technology and biology, which are being investigated to increase the therapeutic ratio of this disease.

mRNA expression of MAGE-A3 gene in leukemia cells.

Leukemia-associated antigens such as proteins encoded by MAGE genes might provide tools for immunotherapy of leukemia. Positive and negative results of MAGE-A gene expression in hematological malignancies have been reported. This led us to study MAGE-A gene expression in human leukemias using RT-PCR. Among 115 leukemias from various subtypes, 14/34 (41.17%) AML were positive for one of the three genes analyzed (MAGE-A1 1/32; MAGE-A3 10/32; MAGE-B2 3/12). Expression was also detected in 23/76 (30.26%) B-cell ALL patients (MAGE-A1 2/53; MAGE-A3 20/53; MAGE-B2 1/32). One of these patients expressed both MAGE-A1 (weak signal) and -A3 (strong signal) genes. Other patient with CML were positive for MAGE-B2 (1/5, 20%). MAGE-A3 expression data were corroborated by real time RT-PCR through determination of MAGE-A3 transcript levels. We concluded that the MAGE-A3 gene is expressed at the mRNA level in a proportion of human leukemias.

The breast dynamic contrast enhanced MRI: preliminary results of a quantitative analysis.

The objective of our study was to apply a quantitative analysis to the dynamic contrast enhanced MR imaging of the breast. Automated criteria increase the objectivity and reproducibility of the diagnostic interpretation of the imaging for differentiating benign and malignant lesions. The validation of this applied method was evaluated by analysing the time- signal intensity curves and the performance of the extracted enhancement parameters. The performance of some extracted parameters was evaluated by ROC (Receiver Operating Characteristic) analysis. These parameters were found to be particularly accurate in differentiating lesions.

Asian-American variants of human papillomavirus 16 and risk for cervical cancer: a case-control study.

BACKGROUND: Human papillomavirus 16 (HPV16) has a number of variants, each with a different geographic distribution and some that are associated more often with invasive neoplasias. We investigated whether the high incidence of cervical cancer in Mexico (50 cases per 100 000 women) may be associated with a high prevalence of oncogenic HPV16 variants. METHODS: Cervical samples were collected from 181 case patients with cervical cancer and from 181 age-matched control subjects, all from Mexico City. HPV16 was detected with an E6/E7 gene-specific polymerase chain reaction, and variant HPV classes and subclasses were identified by sequencing regions of the E6 and L1/MY genes. Clinical data and data on tumor characteristics were also collected. All statistical tests were two-sided. RESULTS: HPV16 was detected in cervical scrapes from 50.8% (92 of 181) of case patients and from 11% (20 of 181) of control subjects. All HPV16-positive samples, except one, contained European (E) or Asian-American (AA) variants. AA and E variants were found statistically significantly more often in case patients (AA = 23.2% [42 of 181]; E = 27.1% [49 of 181]) than in control subjects (AA = 1.1% [two of 181]; E = 10% [18 of 181]) (P<.001 for case versus control subjects for either E or AA variants, chi2 test). However, the frequency of AA variants was 21 times higher in cancer patients than in control subjects, whereas that ratio for E variants was only 2.7 (P =.006, chi2 test). The odds ratio (OR) for cervical cancer associated with AA variants (OR = 27.0; 95% confidence interval [CI] = 6.4 to 113.7) was higher than that associated with E variants (OR = 3.4; 95% CI = 1.9 to 6.0). AA-positive case patients (46.2 +/- 12.5 years [mean +/- standard deviation]) were 7.7 years younger than E-positive case patients (53.9 +/- 12.2 years) (P =.004, Student's t test). AA variants were associated with squamous cell carcinomas and adenocarcinomas, but E variants were associated with only squamous cell carcinomas (P =.014, Fisher's exact test). CONCLUSIONS: The high frequency of HPV16 AA variants, which appear to be more oncogenic than E variants, might contribute to the high incidence of cervical cancer in Mexico.

Cuticular hydrocarbon variability among Triatoma dimidiata (Hemiptera: Reduviidae) populations from Mexico and Guatemala.

The geographic variation in the cuticular hydrocarbon pattern among 11 populations of Triatoma dimidiata Latreille (Hemiptera: Reduviidae: Triatominae) from different regions of Mexico and Guatemala, was studied using capillary gas chromatography. T. dimidiata populations were differentiated based on the relative amounts of 71 hydrocarbon components. Insect population classification was mostly in agreement with their geographical vicinity; Mexican populations from the Yucatan peninsula grouped together with those from northern Guatemala, insects from the Mexican Gulf coast states were closely related to those collected from northern Oaxaca, and to a lesser extent, to insects from Chiapas. Insects from southern Oaxaca were clustered together with those from southern Guatemala. All these populations were clearly separated from Guatemalan specimens collected in caves from Alta Verapaz.

Understanding nutritional interventions and physical exercise in non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in adults and its prevalence is rising around the world. This pathology is characterized by accumulation of liver fat, which exceeds 5% of liver weight in absence of alcohol consumption, viral infection or other hepatic etiology. Since NAFLD has been associated with obesity, insulin resistance, diabetes or alteration of lipid profiles, it is considered as the liver manifestation of metabolic syndrome. Pathogenic mechanisms of NAFLD have not been clearly elucidated, but different events such as lipid accumulation, insulin resistance, oxidative and endoplasmic reticulum stress, mitochondrial dysfunction and inflammation are involved. Modifications in lifestyle constitute the first line for the management of NAFLD. Nutritional interventions include low fat and carbohydrate diet with higher polyunsaturated fatty acids ingestion. Moreover, supplementation with antioxidant and cytoprotective agents could be useful to decrease oxidative stress, inflammation and fibrosis. Physical activity enables to reduce the expression of lipogenic genes, fat accumulation, or insulin resistance and improves cardiorespiratory fitness. Benefits have been found following both aerobic exercise and resistance training, and remain even after exercise cessation. However, more studies are required to analyze the molecular and cellular mechanisms involved in nutritional and physical intervention, and to define the volume of activity required and its association with weight loss. In this paper, we offer an updated overview of the mechanisms implicated in the progression of NAFLD, and analyze the beneficial effects of nutritional interventions and physical exercise in the prevention and treatment of this condition.

Isoenzyme variability of five principal triatomine vector species of Chagas disease in Mexico.

Triatoma barberi, T. dimidiata, T. longipennis, T. pallidipennis and T. picturata, all key Chagas disease vectors in Mexico, were analysed by multilocus enzyme electrophoresis (MLEE) at 17 putative loci. The majority of insect specimens studied were collected from domestic and peridomestic structures from multiple geographic locations while others were collected from sylvatic areas. T. barberi was the least polymorphic species (P(0.95)=0.18), with polymorphism rates of the other species ranging from 0.29 to 0.50. T. barberi, a member of the protracta complex, clustered apart from the other studied species by Nei's genetic distance with >1.36, and at least eight loci were found to be diagnostic for this species. T. dimidiata was more related to T. longipennis, T. pallidipennis and T. picturata (phyllosoma complex) than to T. barberi, with a genetic distance averaging 0.36 with the phyllosoma complex species. In contrast, the genetic distances between the three phyllosoma complex species were not significantly different from zero, and there were no species-specific loci differentiating among them. The results strongly support the grouping of these three species in one complex, separate from the two other species studied.

Peptidergic, catecholaminergic and morphological properties of avian chromaffin cells are modulated distinctively by growth factors.

Most neurons and endocrine cells are known to co-express a 'classical neurotransmitter' with one or more neuropeptides. Although their expression has been shown to be modulated by differentiation factors, it is not known if particular combinations of neurotransmitter/neuropeptide(s) are co-regulated. We have analyzed the effect of nerve growth factor (NGF), neurotrophin-3 (NT-3), brain derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-beta 1) on the modulation of neuroactive substances co-expressed by avian chromaffin cells. The content of the neuropeptides neuropeptide Y (NPY), enkephalin (ENK) and somatostatin (SS) was measured by radioimmunoanalysis, and the content of the catecholamines norepinephrine (NE) and epinephrine (E) by high pressure liquid chromatography-electrochemistry (HPLC-EC). In addition, the morphological differentiation of chromaffin cells in response to the growth factors was assessed. All of the studied factors had distinct effects on the chromaffin content of neuropeptides and catecholamines. Our results show that the modulation of CAs and neuropeptides, and among the neuropeptides themselves is completely dissociated. Moreover, the cellular responses to the different growth factors show that neurochemical properties are modulated independently of morphological ones.

Growth factors effects on the expression of morphological and biochemical properties of avian embryonic sympathetic cells. Emphasis on NGF.

Growth factors are known to be important agents in the differentiation and modulation of neuronal phenotypes. We have analyzed the effect of several growth factors on the modulation of morphological and biochemical properties of avian embryonic sympathetic neurons. The growth factors studied include: nerve growth factor (NGF), neurotrophin-3 (NT-3), brain derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF) and transforming growth factor beta-1 (TGF-beta1). Morphological properties were analyzed by immunocytochemistry to neurofilament proteins and visualization of fibers after glyoxylic acid-induced fluorescence. Biochemical modulation was determined by radioimmunoanalysis for the peptides enkephalin (ENK), somatostatin (SS) and neuropeptide Y (NPY) and by HPLC-electrochemistry quantification of catecholamines. Similar to previous results using chromaffin cell cultures [R. Ramírez-Ordóñez, J.E. García-Arrarás, Peptidergic, catecholaminergic and morphological properties of avian chromaffin cells are modulated distinctively by growth factors, Dev. Brain Res., 87 (1995) 160-171], we found a dissociation in the modulation of biochemical and morphological properties, however, the effect of specific factors differed between the chromaffin and sympathetic cultures. We have focused on NGF to analyze its effect on the sympathetic peptide phenotypes and its lack of an effect on the chromaffin cell peptide phenotypes. The results presented here, establish interesting differences between chromaffin cells and sympathetic neurons that are of importance to studies of cell lineage and differentiation.