Application of a digital PCR method for WT1 to myeloid neoplasms in CR and deep ELN WT1 molecular response (< 10 copies).

Bone marrow WT1 mRNA levels assessed by the ELN method are useful to establish prognostic correlations in myeloid malignancies treated with chemotherapy or hematopoietic stem cell transplantation (HCT). Those patients with WT1 levels below ten copies have a good outcome. However, some of these patients relapse. To further characterize this group of cases, we applied a new and sensitive digital (ddPCR) WT1 method. A consecutive series of 49 patients with treated myeloid malignancies and with an ELN WT1 quantitation of < 10 copies were included in the study. All cases (47 AML and 2 MDS) have received intensive chemotherapy or HCT. One to four micrograms of total RNA were retrotranscribed to obtain ≥ 10,000 ABL1 copies using the ELN protocol. Only those cases with a good quality cDNA were used in the ddPCR WT1 test. The ddPCR Gene Expression WT1 Assay of Bio-Rad© was used to perform the PCR amplification, and the microdroplets were quantified in the Bio-Rad's QX200 droplet reader. Eighteen patients showed a negative WT1 ddPCR assay (0 copies/µl), whereas 31 cases were positive (results ranged from 1 to 15.2 copies/µl). Survival analysis showed statistically significant differences in terms of OS between both groups, 83 ± 8% vs. 46 ± 9% (p = 0.024). A statistically significant correlation was also found between ddPCRWT1 results and CD123+ cell number detected by flow cytometry (p = 0.024). Larger series of patients tested with the current ddPCRWT1 method will solve whether it could be used to stratify patients with myeloid malignancies achieving deep WT1 molecular response (< 10 copies).

A Spanish Society joint SECO and SEEDO approach to the Post-operative management of the patients undergoing surgery for obesity.

PURPOSE: Bariatric surgery is the method of choice for the management or treatment of obesity. Bariatric surgery brings about several physiological changes in the body and is associated with set of complications. The aim of this study is to provide guidelines on post bariatric surgery management based on consensus by the Spanish society for Obesity Surgery (Sociedad Española de Cirugía de la Obesidad) (SECO) and the Spanish Society for the Study of Obesity (Sociedad Española para el Estudio de la Obesidad) (SEEDO). METHOD: The boards proposed seven experts from each society. The experts provided the evidence and a grade of recommendation on the selected topics based on systematic reviews/meta-analysis. A list of clinical practical recommendations levels of evidence and grades of these recommendations was derived from the consensus statements from the members of these societies. RESULTS: Seventeen topics related to post-operative management were reviewed after bariatric surgery. The experts came with 47 recommendations and statements. The mean number of persons voting at each statement was 54 (range 36-76). CONCLUSION: In this consensus, we have designed a set of guidelines to be followed while managing patients after bariatric surgery. Expertise and knowledge of the clinicians are required to convey suitable considerations to the post-bariatric patients. There should also be extensive follow-up plans for the bariatric surgery patients.

Incidence of new-onset benign anal disorders after bariatric surgery.

New-onset benign anal disorders (NBADs) represent a potential complication following bariatric surgery, although their incidence in this population is not well studied. The preoperative characteristics, weight loss, bowel habits and NBADs data of 196 patients with bariatric surgery were collected by telephone interviews and medical records review and evaluated retrospectively. Ninety-nine patients had undergone gastric bypass (GBP) and 97 had a modified biliopancreatic diversion (MBPD). Fifty-nine patients were excluded. The mean follow-up of the remaining 137 patients was 87.8 months, and 51 of them (37.2%) developed NBADs. Haemorrhoids were the most common diagnosis and 27.5% of the patients that developed NBADs were treated surgically. Patients who developed NBADs had a longer follow-up time (92.5 vs. 85.1 months, P = 0.003), and those with an abnormal bowel habit (diarrhoea or constipation) had a higher percentage of NBADs (54.5 vs. 28.3%, P = 0.003). NBADs were more frequent after MBPD (52.9%) than after GBP (21.7%) (P < 0.001). Multivariate analysis found that only an abnormal bowel habit was associated with NBADs, with an odds ratio of 3.2 (95% CI: 1.5-6.9, P = 0.003). As NBADs are a common complication after bariatric surgery, further studies should be performed to find the reasons for these complications.

Evaluation of water quality at the source of streams of the Sinos River Basin, southern Brazil / Avaliacão da qualidade da água das nascentes de arroios da Bacia do Rio dos Sinos, região sul do Brasil

The Sinos River Basin (SRB) is located in the northeastern region of the state of Rio Grande do Sul (29º20' to 30º10'S and 50º15' to 51º20'W), southern Brazil, and covers two geomorphologic provinces: the southern plateau and the central depression. It is part of the Guaíba basin, has an area of approximately 800 km 2 and contains 32 counties. The basin provides drinking water for 1.6 million inhabitants in one of the most important industrial centres in Brazil. This study describes different water quality indices (WQI) used for the sub-basins of three important streams in the SRB: Pampa, Estância Velha/Portão and Schmidt streams. Physical, chemical and microbiological parameters assessed bimonthly using samples collected at each stream source were used to calculate the Horton Index (HI), the Dinius Index (DI) and the water quality index adopted by the US National Sanitation Foundation (NSF WQI) in the additive and multiplicative forms. These indices describe mean water quality levels at the streams sources. The results obtained for these 3 indexes showed a worrying scenario in which water quality has already been negatively affected at the sites where three important sub-basins in the Sinos River Basin begin to form.

A Bacia Hidrográfica do Rio dos Sinos (BHRS) está localizada na região Nordeste do Estado do Rio Grande do Sul (29º20' to 30º10'S and 50º15' to 51º20'W), na região Sul do Brasil, envolvendo duas regiões geomorfológicas, o planalto Sul e a depressão central. Esta bacia faz parte da Bacia do Guaíba e tem uma área de aproximadamente 800 km 2, atingindo 32 municípios. A BHRS fornece água de abastecimento para cerca de 1.6 milhões de habitantes e para um dos mais importantes centros industriais do Brasil. Este artigo apresenta a determinação de diferentes índices de qualidade de água (IQA) para as regiões de nascente de três importantes sub-bacias pertencentes à BHRS: arroios Pampa, Estância Velha/Portão e Schmidt. Utilizando parâmetros físicos, químicos e microbiológicos, analisados em amostras coletadas bimestralmente em cada ponto próximo à nascente, foram calculados os índices de Horton (IH), de Dinius (ID) e o índice de qualidade de água da National Sanitation Foundation, EUA (IQANSF) usando os métodos somatório e produtório. Os resultados foram analisados para verificação da qualidade da água nos pontos amostrais a fim de comparar e classificar a água na região das nascentes das três sub-bacias. Uma avaliação conjunta dos índices calculados indicou que a água nos três pontos amostrais apresenta qualidade média. Os resultados dos três índices convergem a um cenário preocupante que indica que, desde os pontos iniciais de formação de três importantes sub-bacias do rio dos Sinos, já ocorrem problemas de qualidade de água.

Mild caloric restriction reduces blood pressure and activates endothelial AMPK-PI3K-Akt-eNOS pathway in obese Zucker rats.

Genetic obesity models exhibit endothelial dysfunction associated to adenosine monophosphate-activated protein kinase (AMPK) dysregulation. This study aims to assess if mild short-term caloric restriction (CR) restores endothelial AMPK activity leading to an improvement in endothelial function. Twelve-week old Zucker lean and obese (fa/fa) male rats had access to standard chow either ad libitum (AL, n=8) or 80% of AL (CR, n=8) for two weeks. Systolic blood pressure was significantly higher in fa/fa AL rats versus lean AL animals, but was normalized by CR. Endothelium-dependent relaxation to acetylcholine (ACh, 10(-9) to 10(-4) M) was reduced in fa/fa AL compared to control lean AL rats (p<0.001), and restored by CR. The AMPK activator AICAR (10(-5) to 8·10(-3) M) elicited a lower relaxation in fa/fa AL rings that was normalized by CR (p<0.001). Inhibition of PI3K (wortmannin, 10(-7) M), Akt (triciribine, 10(-5) M), or eNOS (L-NAME, 10(-4) M) markedly reduced AICAR-induced relaxation in lean AL, but not in fa/fa AL rats. These inhibitions were restored by CR in Zucker fa/fa rings. These data show that mild short-term CR improves endothelial function and lowers blood pressure in obesity due to the activation of the AMPK-PI3K-Akt-eNOS pathway.

Effects of nickel on the fern Regnellidium diphyllum Lindm. (Marsileaceae).

The heterosporous fern Regnellidium diphyllum occurs in southern Brazil and some adjoining localities in Uruguay and Argentina. Currently it is on the list of threatened species in the state of Rio Grande do Sul. Anthropic alterations such as the conversion of wetlands into agricultural areas or water and soil contamination by pollutants may compromise the establishment and survival of this species. Nickel (Ni) is an essential nutrient for plants but increasing levels of this metal due to pollution can cause deleterious effects especially in aquatic macrophytes. Megaspore germination tests were performed using Meyer's solution, at concentrations of 0 (control), 0.05, 0.5, 1, 5, 10, 20, 30, 50 and 100 mg L(-1) of Ni. The initial development of apomictic sporophytes was studied using solutions containing 0 (control) to 4.8 mg L(-1) of Ni. A significant negative relation was observed between the different Ni concentrations and the megaspore germination/sporophyte formation rates. Primary roots, primary leaves and secondary leaves were significantly shorter at 3.2 and 4.8 mg L(-1) of Ni, when compared with the treatment without this metal. At 4.8 mg L(-1), leaves also presented chlorosis and necrosis. The introduction of pollutants with Ni in the natural habitat of Regnellidium diphyllum may inhibit the establishment of plants in the initial stage of development, a problem to be considered in relation to the conservation of this species.

[Indicators in primary care: reality or desire. SESPAS report 2012]. / Indicadores en atención primaria: la realidad o el deseo. Informe SESPAS 2012.

The aim of this article is two-fold: firstly, to illustrate international experiences of assessing primary care performance by using examples of both cross-country comparisons and national assessment efforts and, secondly, to describe the progress achieved to date in the articulation of the Spanish Primary Care Information System. The central role assigned to primary care in conceptual frameworks clashes with the severe limitations in the quality and availability of the data required to construct the indicators. The lack of systematic and standardized databases covering diagnosis, procedures and intermediate results in individual patients is endemic across countries. Filling this gap has become a priority in most countries, especially during the last decade. In Spain, the concept of the Primary Care Information System appeared in 2003 and the first data/results were published in 2006. Since then, distinct elements have been progressively incorporated with the involvement of the various autonomous regions in Spain. Currently, the system includes various aspects such as the population assigned, the available resources, a national catalogue of primary care centers, and activity. The next challenge in this work in progress is to build a database of clinical information.

Germination and sporophytic development of Regnellidium diphyllum Lindman (Marsileaceae) in the presence of hexavalent chromium.

Regnellidium diphyllum Lindman is a heterosporous fern, growing in aquatic environments and surrounding wetlands, which is assumed to be threatened by increasing water pollution and disappearance of its natural habitats. Among contaminants, hexavalent chromium - Cr(VI) - is known to be present in effluents from some leather tanning factories. Megaspore germination tests were performed using Meyer's solution, at concentrations 0 (control), 0.1, 0.5, 1, 5, 10, 15, 20, 30, 50, and 80 mg.L⁻¹, from a standard solution of Titrisol® 1000 mg.L⁻¹. The primary development of apomictic sporophytes was studied using solutions containing 0.025 to 4.8 mg.L⁻¹ of Cr(VI). The experiments were conducted in a growth chamber at 24 ± 1 ºC and for a 12-hour photoperiod under fluorescent lights, providing a nominal irradiance of 77 µmol.m⁻²/s. Significant differences in megaspore germination, with subsequent sporophytic development, were verified from 0.5 mg.L⁻¹ Cr(VI) concentration onwards. Growth of primary root and primary and secondary leaves was significantly reduced at 3.2 mg.L⁻¹ Cr(VI) concentration or higher. Considering the pollution from Cr(VI) in some areas of R. diphyllum natural occurrence, these data indicate that low reproductive rates and disappearance of populations are likely to occur in these situations.

Effects of hTERT on genomic instability caused by either metal or radiation or combined exposure.

Genomic instability is considered to be an important component in carcinogenesis. It can be caused by low-dose exposure to agents, which appear to act through induction of stress-response pathways related to oxidative stress. These agents have been studied mostly in the radiation field but evidence is accumulating that chemicals, especially heavy metals such as Cr (VI), can also act in the same manner. Previous work showed that metal ions could initiate long-term genomic instability in human primary fibroblasts and this phenomenon was regulated by telomerase. The aim of this study was to examine the difference in clonogenic survival and cytogenetic damage after exposure to Cr (VI) and radiation both singly and in combination in normal human fibroblasts (hTERT- cells) and engineered human fibroblasts, infected with a retrovirus carrying a cDNA encoding hTERT, which rendered these cells telomerase positive and replicatively immortal (hTERT+ cells). Cr (VI) induced genomic instability in hTERT- cells but not in hTERT+ cells, whereas radiation induced genomic instability in hTERT+ cells and to a lesser extent in hTERT- cells. Combined exposure caused genomic instability in both types of cells. However, this genomic instability was more pronounced in hTERT- cells after radiation followed by Cr (VI) and more pronounced in hTERT+ cells after Cr (VI) followed by radiation. Moreover, the biological effects provoked by combined exposure of Cr (VI) and radiation also led to a synergistic action in both types of cells, compared to either Cr (VI) treatment only or radiation exposure only. This study suggests that telomerase can prevent genomic instability caused by Cr (VI), but not by radiation. Furthermore, genomic instability may be prevented by telomerase when cells are exposed to radiation and then Cr (VI) but not after exposure to Cr (VI) and then radiation.

[Nutritional implications of bariatric surgery on the gastrointestinal tract]. / Implicaciones nutricionales de la cirugía bariátrica sobre el tracto gastrointestinal.

Anatomical change in the anatomy of the gastrointestinal tract after bariatric surgery leads to modification of dietary patterns that have to be adapted to new physiological conditions, either related with the volume of intakes or the characteristics of the macro- and micronutrients to be administered. Restrictive diet after bariatric surgery (basically gastric bypass and restrictive procedures) is done at several steps. The first phase after surgery consists in the administration of clear liquids for 2-3 days, followed by completely low-fat and high-protein content (> 50-60 g/day) liquid diet for 2-4 weeks, normally by means of formula-diets. Soft or grinded diet including very soft protein-rich foods, such as egg, low-calories cheese, and lean meats such as chicken, cow, pork, or fish (red meats are not so well tolerated) is recommended 2-4 weeks after hospital discharge. Normal diet may be started within 8 weeks from surgery or even later. It is important to incorporate hyperproteic foods with each meal, such egg whites, lean meats, cheese or milk. All these indications should be done under the supervision of an expert nutrition professional to always advise the patients and adapting the diet to some special situations (nausea/vomiting, constipation, diarrhea, dumping syndrome, dehydration, food intolerances, overfeeding, etc.). The most frequent vitamin and mineral deficiencies in the different types of surgeries are reviewed, with a special focus on iron, vitamin B12, calcium, and vitamin D metabolism. It should not be forgotten that the aim of obesity surgery is making the patient loose weight and thus post-surgery diet is designed to achieve that goal although without forgetting the essential role that nutritional education has on the learning of new dietary habits contributing to maintain that weight loss over time.

Effects of hTERT on metal ion-induced genomic instability.

There is currently a great interest in delayed chromosomal and other damaging effects of low-dose exposure to a variety of pollutants which appear collectively to act through induction of stress-response pathways related to oxidative stress and ageing. These have been studied mostly in the radiation field but evidence is accumulating that the mechanisms can also be triggered by chemicals, especially heavy metals. Humans are exposed to metals, including chromium (Cr) (VI) and vanadium (V) (V), from the environment, industry and surgical implants. Thus, the impact of low-dose stress responses may be larger than expected from individual toxicity projections. In this study, a short (24 h) exposure of human fibroblasts to low doses of Cr (VI) and V (V) caused both acute chromosome damage and genomic instability in the progeny of exposed cells for at least 30 days after exposure. Acutely, Cr (VI) caused chromatid breaks without aneuploidy while V (V) caused aneuploidy without chromatid breaks. The longer-term genomic instability was similar but depended on hTERT positivity. In telomerase-negative hTERT- cells, Cr (VI) and V (V) caused a long lasting and transmissible induction of dicentric chromosomes, nucleoplasmic bridges, micronuclei and aneuploidy. There was also a long term and transmissible reduction of clonogenic survival, with an increased beta-galactosidase staining and apoptosis. This instability was not present in telomerase-positive hTERT+ cells. In contrast, in hTERT+ cells the metals caused a persistent induction of tetraploidy, which was not noted in hTERT- cells. The growth and survival of both metal-exposed hTERT+ and hTERT- cells differed if they were cultured at subconfluent levels or plated out as colonies. Genomic instability is considered to be a driving force towards cancer. This study suggests that the type of genomic instability in human cells may depend critically on whether they are telomerase-positive or -negative and that their sensitivities to metals could depend on whether they are clustered or diffuse.

Severe hypocalcemia following total thyroidectomy after biliopancreatic diversion.

Patients undergoing malabsoprtive operations for bariatric surgery are prone to disturbances of bone metabolism, but this does not commonly lead to clinical symptoms. We present a morbidly obese patient who had undergone the biliopancreatic diversion of Larrad, and presented clinical symptoms of severe hypocalcemia and tetany after total thyroidectomy. Very high doses of i.v. calcium and calcitriol and 10 days of hospitalization were required to control the symptoms and correct plasma levels. The physiological aspects that contribute to this situation are discussed.

[Implications of fiber in different pathologies]. / Implicaciones de la fibra en distintas patologías.

Three decades ago, the observations of Trowell and Burkitt gave rise to the "fibre theory", in which it was contended that there was a link between the consumption of a diet rich in fibre and non-processed carbohydrates and the level of protection against many of the "first world diseases" such as constipation, diverticulosis, cancer of the colon, diabetes, obesity and cardiovascular disease. Since that time, numerous studies have been presented to analyze the relationship between fibre and disorders of the gastrointestinal tract and other processes with severe health implications. The present revision looks at the experience accumulated over this period regarding the importance of the consumption of fibre for certain phatologies. It not only deals with the epidemiological relationship existing between fibre intake and the development of diseases such as cancer of the colon or cardiovascular disorders but also reviews the interest of fibre a therapeutic agent, in view of the current information available on its different mechanism of action. Thus the possibility of using soluble fibre has taken on renewed interest for the treatment of inflammatory intestinal disease, for control of diarrhoea, in irritable bowel syndrome or no modulate the concentrations of glycaemia or cholesterol. Three is a discussion of the discrepancies found between the consumption of fibre and diverticular disease, the treatment of constipation and the association with obesity and cardiovascular disease. Despite the accumulated evidence on the consumption of fibre, there is currently no consensus as to recommendations on what type of fibre and the optimal amount that should be consumed. A high fibre intake (> 25-30 g/day) based on a variety of food sources (fruit, vegetable, legumes, cereals) is the only way to avoid many of the disorders mentioned. The consumption of a particular type of fibre (soluble or insoluble) is limited to the treatment of certain processes, because its individual relationship with many disorders is still pending determination.

[Clinical features and weight loss in patients with morbid obesity after bariatric surgery]. / Características clínicas y pérdida de peso en pacientes con obesidad mórbida tras cirugía bariátrica.

OBJECTIVE: To analyze the psychological status of morbidly obese patients suffering changes after surgical procedure for weight reduction. METHODS: We analyze the weight loss in 100 morbidly obese patients (85 female, 15 male) after bariatric surgery (vertical banded gastroplasty). We correlated the percentage of weight loss with the age of these patients, initial weight (body mass index at surgery), quality of life, eating behaviour, psychopathology, personality, and marital status. RESULTS: We have found relation between initial weight and weight loss (heavier patients lose more weight). The percentage of weight loss is associated with the Eating Behaviour (Body Dissatisfaction, Disinhibition), the Quality of Life (Positive Affect), the Psychopathology (Hostility), the Personality Traits (Narcissist), and the Marital Status (Idealistic Distortion). CONCLUSIONS: The weight loss in morbidly obese patients after surgery is major in the more obese one. The satisfactory weight loss after surgery improve the eating behaviour, quality of life, psychological and marital status.

Eating behavior in morbidly obese patients undergoing gastric surgery: differences between obese people with and without psychiatric disorders.

BACKGROUND: This study analyzes eating behavior in a group of morbidly obese patients who have undergone gastric reduction surgery for weight loss, and evaluates whether the existence of psychiatric comorbidity marks significant differences in their eating behavior. METHODS: The study group was composed of 100 morbidly obese patients (85 females, 15 males) who had received surgical treatment for weight reduction (vertical banded gastroplasty). 40 of these patients (40%) met ICD-10 criteria for the diagnosis of psychiatric disorders and were included in the "Psychiatric Obese group" (PO). The other 60 patients (60%) did not show ICD-10 diagnostic criteria and were included in the "Non-Psychiatric Obese group" (NO). Each patients completed the Binge Eating Scale (BES), the Three Factor Eating Questionnaire, the Bulimia Investigatory Test-Edinburgh (BITE), and the Eating Disorder Inventory (EDI). RESULTS: Significant differences were found between the two groups (PO and NO) in the Binge Eating Scale (p < 0.001), Three Factor Eating Questionnaire subscale Disinhibition (p < 0.001), BITE (p < 0.001), Eating Disorder Inventory subscale Perfectionism (p < 0.002), and Global EDI (p < 0.001). Logistic regression analysis showed correlation between PO group and Global EDI (Odds Ratio OR = 1.43) and BITE (OR = 1.16). No significant gender differences were found for eating behavior, clinical diagnosis, age, percentage of weight loss, time after operation, and BMI before surgery. CONCLUSION: Surgically treated morbidly obese patients with a psychiatric disorder (PO) have a more destructured eating pattern (with a predominance of binge eating and disinhibition) than NO.

Selective importation of RNA into isolated mitochondria from Leishmania tarentolae.

All mitochondrial tRNAs in kinetoplastid protozoa are encoded in the nucleus and imported from the cytosol. Incubation of two in vitro-transcribed tRNAs, tRNA(Ile)(UAU) and tRNA(Gln)(CUG), with isolated mitochondria from Leishmania tarentolae, in the absence of any added cytosolic fraction, resulted in a protease-sensitive, ATP-dependent importation, as measured by nuclease protection. Evidence that nuclease protection represents importation was obtained by the finding that Bacillus subtilis pre-tRNA(Asp) was protected from nuclease digestion and was also cleaved by an intramitochondrial RNase P-like activity to produce the mature tRNA. The presence of a membrane potential is not required for in vitro importation. A variety of small synthetic RNAs were also found to be efficiently imported in vitro. The data suggest that there is a structural requirement for importation of RNAs greater than approximately 17 nt, and that smaller RNAs are apparently nonspecifically imported. The signals for importation of folded RNAs have not been determined, but the specificity of the process was illustrated by the higher saturation level of importation of the mainly mitochondria-localized tRNA(Ile) as compared to the level of importation of the mainly cytosol-localized tRNA(Gln). Furthermore, exchanging the D-arm between the tRNA(Ile) and the tRNA(Gln) resulted in a reversal of the in vitro importation behavior and this could also be interpreted in terms of tertiary structure specificity.

The pur7 gene from the puromycin biosynthetic pur cluster of Streptomyces alboniger encodes a nudix hydrolase.

Pur7 is the product of a gene from the puromycin biosynthetic pur cluster of Streptomyces alboniger. It was expressed in Escherichia coli as a recombinant protein fused to a His tag and then was highly purified through a Ni(2+) column. It showed a 3'-amino-3'-dATP pyrophosphohydrolase (nudix) activity which produced 3'-amino-3'-dAMP and pyrophosphate. This is consistent with the presence of a nudix box in its amino acid sequence. As observed with other nudix hydrolases, Pur7 has an alkaline pH optimum and a requirement for Mg(2+). Among a large variety of other nucleotides tested, only 3'-amino-3'-dTTP was a Pur7 substrate, although at lower reaction rates than 3'-amino-3'-dATP. These findings suggest that Pur7 has a high specificity for the 3' amino group at the ribofuranoside moiety of these two substrates. The K(m) and V(max) values for these dATP and dTTP derivatives were 120 microM and 17 microM/min and 3.45 mM and 12.5 microM/min, respectively. Since it is well known that 3'-amino-3'-dATP is a strong inhibitor of DNA-dependent RNA polymerase, whereas 3'-amino-3'-dAMP is not, Pur7 appears to be similar to other nudix enzymes in terms of being a housecleaning agent that permits puromycin biosynthesis to proceed through nontoxic intermediates. Finally, the identification of this activity has allowed a revision of the previously proposed puromycin biosynthetic pathway.