Serum profiling and biomarker discovery of rat mammary tumors using mass-coded abundance tags (MCAT)

Advances in Mass-spectrometry techniques allow for the rapid processing and evaluation of complex biological mixtures such as blood/serum. These samples represent a protein rich environment as well as a sentinel monitoring system of the entire organism. The central tenet of these studies is that changes in the microenvironment of a tissue, brought about by a disease process, will lead to sufficient changes in the protein and peptide pattern of the serum, such that the differences can be accurately detected and correctly associated with a particular disease state. Using mass-spectrometry approaches we have developed techniques that allow us to compare samples from tumor-free and tumor present serum samples simultaneously to find biomarkers that indicate the presence of cancer. To examine potentially important but less abundant proteins, ultrafiltration (UF) was used to eliminate the more abundant proteins and combine this with the non-isotopic peptide tags (S-methylthioacetimidate and S-methyl thiopropionimidate) described by Beardsley and Reilley (J. Proteome Res. 2: 15-21, 2003) to differentiate our samples. Use of these mass-coded abundance tags (MCAT) allows for simultaneous evaluation of serum samples from tumor present, and tumor free animals. Using an oa time-of-flight mass-spectrometer (Q-tof) with electrospray ionization we produce high quality spectrums to screen for peptides that have only one tag. Specificity of tagging increases the likelihood that the peptide resulted from a protein unique to either the control or conditioned state. Using the ms/ms function of the Q-tof we sequence the peptide and identify the parent protein. Specifically, our lab is using UF, MCAT and the Q-tof to evaluate rat models of chemically-induced tumors. By using animal models we overcome much of the variability that may exist in human serum samples due to differences in gender, diet and cancer initiation. We have shown that these systems allow for the identification of both small molecules such as Alpha S1 casein precursor (24 kDa) as well as proteins greater than the MCO such as Fibrinogen alpha/alpha E precursor and Coagulation factor 2 (86 and 70 kDa, respectively). With positive sequence identification we can now evaluate the tumors themselves to determine if the proteins are over-expressed in the tumor vs. normal tissues. Using this method of “bottom-up” analysis provides information on the nature and composition of our samples to more rapidly identify those proteins that are unique to the tumor state of the animals

Resveratrol, but not EGCG, in the diet suppresses DMBA-induced mammary cancer in rats

Despite the advent of new and aggressive therapeutics, breast cancer remains a leading killer among women; hence there is a need for the prevention of this disease. Several naturally occurring polyphenols have received much attention for their health benefits, including anti-carcinogenic properties. Two of these are resveratrol, a component of red grapes, and epigallocatechin-3-gallate (EGCG), the major catechin found in green tea. In this study, we tested the hypothesis that these two polyphenols protect against chemically-induced mammary cancer by modulating mammary gland architecture, cell proliferation, and apoptosis. Female Sprague-Dawley CD rats were exposed to either resveratrol (1 g/kg AIN-76A diet), EGCG (0.065% in the drinking water), or control diet (AIN-76A) for the entirety of their life starting at birth. At 50 days postpartum, rats were treated with 60 mg dimethylbenz[a]anthracene (DMBA)/kg body weight to induce mammary cancer. Resveratrol, but not EGCG, suppressed mammary carcinogenesis (fewer tumors per rat and longer tumor latency). Analysis of mammary whole mounts from 50-day-old rats revealed that resveratrol, but not EGCG, treatment resulted in more differentiated lobular structures. Bromodeoxyuridine (BrdU) incorporation studies showed that resveratrol treatment caused a significant reduction in proliferative cells in mammary terminal ductal structures at 50 days postpartum, making them less susceptible to carcinogen insult. The epithelial cells of terminal end buds in the mammary glands of resveratrol-treated rats also showed an increase in apoptotic cells compared to the control or EGCG-treated rats as measured by a DNA fragmentation assay. At the given doses, resveratrol treatment resulted in a serum resveratrol concentration of 2.00 μM, while treatment with EGCG resulted in a serum EGCG concentration of 31.06 nM. 17β-Estradiol, progesterone, and prolactin concentrations in the serum were not significantly affected by resveratrol or EGCG. Neither polyphenol treatment resulted in toxicity as tested by alterations in body weights, diet and drink consumptions, and day to vaginal opening. We conclude that resveratrol in the diet can reduce susceptibility to mammary cancer, while EGCG in the drinking water at the dose used was not effective

Genistein chemoprevention of prostate cancer in TRAMP mice

Epidemiological studies suggest an inverse association between soy intake and prostate cancer risk. Genistein, the predominant phytoestrogen in soy food, has been proposed as a potential chemopreventive agent due to its anti-estrogen and tyrosine kinase inhibitory effects. To determine the most effective period for genistein chemoprevention, the Transgenic adenocarcinoma mouse prostate (TRAMP) model was used. The treatments were 250 mg genistein/kg AIN-76A diet 1) prepubertally only, 2) in adulthood only or 3) through out life. Controls received AIN-76A diet. By 28 weeks of age, 100% TRAMP mice fed control diet developed prostatic intraepithelial neoplasia (PIN) or adenocarcinomas with 6%, 16%, 44% and 34% developing high grade PIN, well differentiated, moderately differentiated and poorly differentiated prostatic adenocarcinomas, respectively. Prepubertal only (1–35 days postpartum) and adult only genistein treatments (12 – 28 weeks) resulted in 6% and 29% decreases in poorly-differentiated cancerous lesions compared with controls, respectively. The most significant effect was seen in the TRAMP mice exposed to genistein throughout life (1–28 weeks) with a 50% decrease in poorly-differentiated cancerous lesions. In a separate experiment in castrated TRAMP mice, dietary genistein suppressed the development of advanced prostate cancer by 35% compared with controls. Of the tumors that developed in castrated TRAMP mice, 100% were poorly-differentiated in contrast to the 37% of noncastrated TRAMP mice that developed poorly-differentiated tumors. ICI 182,780 (ICI), genistein and estrogen down-regulated androgen receptor (AR), estrogen receptor alpha (ER-α) and progesterone receptor (PR) in the prostates of C57BL/6 mice, and act independently of ER. Our data obtained in intact and castrated transgenic mice suggest that genistein may be a promising chemopreventive agent against androgen-dependent and independent prostate cancers

Three Attempts to Determine the Effects of an Appetitive Secondary Reinforcer on the Extinction of an Avoidance Response

A study by Pavlov (5, p. 30) indicated that normally painful stimuli applied while hungry dogs were feeding produced less overall avoidance behavior than at times when they were not feeding. In a still earlier study, Jones (3) reported that extinction of avoidance to a feared stimulus was hastened by gradually introducing the stimulus while the child subject was eating. More recently, Farber (1) has presented evidence that feeding in the presence of anxiety-producing cues hastened later extinction to those cues as compared to control animals which did not receive the feeding experience. In Miller\u27s words (4): Eating and the emotional responses that accompany it are apparently incompatible with fear, and the attaching of these responses to the stimuli that arouse the fear suppresses it

Limitações da fertilidade do solo de áreas com capoeira e sistemas agroflorestais no Assentamento Tarumã-Mirim (Manaus, AM).

Este trabalho teve como objetivo avaliar os limitantes químicos de qualidade do solo, em capoeiras e sistemas agroflorestais (com média a alta diversidade) de pequenas propriedades agrícolas localizadas no Assentamento Tarumã-Mirim, zona rural de Manaus

A systematic review and meta-synthesis of patients’ experiences and perceptions of seeking and using benzodiazepines and z-drugs: towards safer prescribing

Background Benzodiazepines and Z-drugs are used to treat complaints like insomnia, anxiety and pain. These drugs are recommended for short-term use only, but many studies report long-term use, particularly in older people. Objective The aim of this study was to identify and synthesise qualitative studies exploring patients’ experiences and perceptions of receiving benzodiazepines and Z-drugs, and through this identify factors which perpetuate use of these drugs, and strategies for achieving safer prescribing. Methods A systematic search of six databases for qualitative studies exploring patients’ experiences and perceptions of primary care benzodiazepine and z-drug prescribing published between January 2000 and April 2014 in a European language, and conducted in Europe, the United States, Australia or New Zealand. Reference lists of included papers were also searched. Study quality was assessed using the Critical Appraisal Skills Programme qualitative checklist. Findings were synthesised using thematic synthesis. Results Nine papers were included and seven analytical themes were identified relating to patients’ experiences and perceptions and, within that, strategies for safer prescribing of benzodiazepines and Z-drugs: (1) patients’ negative perceptions of insomnia and its impact, (2) failed self-care strategies, (3) triggers to medical help-seeking, (4) attitudes towards treatment options and service provision, (5) varying patterns of use, (6) withdrawal, (7) reasons for initial or ongoing use. Conclusions Inappropriate use and prescribing of benzodiazepines and Z-drugs is perpetuated by psychological dependence, absence of support and patients’ denial/lack of knowledge of side effects. Education strategies, increased availability of alternatives, and targeted extended dialogue with patients could support safer prescribing

Comparison of a Lightweight Experimental Shaker and an Orchard Tractor Mounted Trunk Shaker for Fresh Market Citrus Harvesting

[EN] A designed lightweight experimental shaker successfully used to collect ornamental oranges has been tested to harvest fresh market citrus. The aim of this study was to evaluate the removal efficiency and operational times of this experimental device compared to an orchard trunk shaker. Three different collecting systems were studied. 'Caracara' citrus trees were tested. Removal efficiency, vibration parameters, fruit and tree damages, and fruit quality were measured. A high-speed camera was used to record operational times and determine cumulative removal percentage over vibration time. The canvases on the ground reduced the severe fruit damages but were not useful to protect against light damages. The experimental shaker produced a higher percentage of slightly damaged oranges. No significant differences in removal efficiency were found between the two harvesting systems. However, removal efficiency using the experimental device could be reduced by 40 percent and working time increase by more than 50 percent when access to the main branches was difficult. In agreement with previous results, the curve representing the branch cumulative removal percentage in time followed a sigmoidal pattern. A model was built showing that during the first 5 s more than 50 percent of the fruits were detachedThis research has been fund by the European Agricultural Fund for Rural Development and cofounded by the Ministerio de Agricultura, Pesca y Alimentacion (project GO "Avances tecnologicos para la modernizacion y la sostenibilidad en la produccion de citricos CITRUSTECH").Ortiz Sánchez, MC.; Torregrosa, A.; Castro-García, S. (2021). Comparison of a Lightweight Experimental Shaker and an Orchard Tractor Mounted Trunk Shaker for Fresh Market Citrus Harvesting. Agriculture. 11(11):1-10. https://doi.org/10.3390/agriculture11111092110111

Evaluation of Citriculture Mechanisation Level in Valencia Region (Spain): Poll Results

[EN] The increase of the technology level of citrus production operations is required to improve production profitability and reduce production costs. In the framework of the project CITRUSTECH ("Technological advances for modernisation and sustainability in citrus production"), three different poll questionnaires were developed and conducted in the Valencia region to assess the citriculture mechanisation level. In total, 142 questionaries for small and medium-size plantations, 32 for cooperative technicians and 16 for large-size plantations were conducted. From a socioeconomic point of view, clear age and sex inequalities were found. From the technological point of view, relevant differences were found between plantation sizes. The role of the cooperative mechanisation services (custom cost) and other customer services was revealed, with a higher percentage of the area under cultivation at the expense of the small-size plantations. The use of some manual tools was confirmed in pruning, even in large-size orchards. In small-size orchards, the use of backpack sprayers was verified. Regarding farm machinery, besides tractors, hydro-pneumatic sprayers and agricultural shredders were employed. No farm machinery was used during harvesting operations, apart from transport equipment, due to the reduced plantation frames.This research has been funded by the European Agricultural Fund for Rural Development and cofounded by the Ministerio de Agricultura, Pesca y Alimentacion (project GO "Avances tecnologicos para la modernizacion y la sostenibilidad en la produccion de citricos CITRUSTECH").Ortiz Sánchez, MC.; Torregrosa, A.; Martínez, JM. (2022). Evaluation of Citriculture Mechanisation Level in Valencia Region (Spain): Poll Results. Agronomy. 12(6):1-6. https://doi.org/10.3390/agronomy120613661612

Conflitos na Área de Preservação Ambiental Tarumã-Açu/Tarumã-Mirim: proteger para quem?.

Conflitos em Áreas de Proteção Ambiental (APAs) revelam a falta de elo entre os mecanismos de gestão ambiental, as comunidades envolvidas e as políticas governamentais. Neste trabalho, são apresentadas as contradições entre os objetivos da criação de uma APA e as atividades, de alto impacto ambiental negativo, realizadas pelas comunidades na APA Tarumã-Açu/Tarumã-Mirim. Compatibilizar o uso sustentável dos recursos naturais com a presença humana exige esforços conjuntos. O não envolvimento da população que vive na área pode acarretar problemas ambientais irreversíveis. Palavras-chave: Conflitos, Área de Proteção Ambiental, Comunidades Rurais.Resumo 10596