Exopolymeric substances from drinking water biofilms: Dynamics of production and relation with disinfection by products.

Exopolymeric substances (EPS) as an external matrix of biofilm could react with disinfectants in drinking water networks forming disinfection by-products (DBP). Based on an experimental setup using two chlorine conditions-biofilm 1 (2.6 ± 0.8 mgCl/L) and biofilm 2 (0.7 ± 0.2 mg Cl/L)-samples of biofilms were recovered during 9 campaigns and EPS were extracted. Analyses of SUVA, fluorescence and amino acid (AA) content were carried out on the EPS to observe variation over time and correlations with DBP formation potential (DBPfp) after chlorination. SUVA values were under 2 L/mgC*m showing that both EPS were hydrophilic. Slightly higher SUVA in biofilm 2 with low variation over time was observed. Fluorescence showed that aromatic proteins and fulvic like substances were the principal components and increased in biofilm 1 over time. AA decreased with time, and higher values of alanine, threonine, proline and isoleucine were observed in biofilm 2. Based on general associations, the SUVA of biofilm 2 correlated well with chloroform (CF) (r = 0.80). Generally, in both biofilms, tryptophan-like substances were negatively correlated with DBP while humic acid-like substances correlated positively, but with low indexes (r = 0.3-0.6). Correlations of data from individual sampling increased the indices (r over 0.8), suggesting a temporal influence of other factors on DBPfp such as inorganics, filtered water and the structural composition of EPS. In biofilm 1, Br-haloacetic acids (Br-HAA), dibromoacetonitrile and bromochloro acetonitrile were inversely associated with arginine and valine, as were di and trichloropropanone to arginine. On the contrary, in biofilm 2, the following amino acids correlated positively with DBP: alanine with Br-HAA, alanine with CF, alanine with N-DBP (chloropicrin, di and tri-chloro acetonitrile), and valine with CF. As this is the first report about the relation between temporal variation of EPS and DBPfp of biofilms in two different chlorinated conditions, it provides new evidence about the function of these complex substances in drinking water systems.

Dynamic Subcellular Localization of Iron during Embryo Development in Brassicaceae Seeds.

Iron is an essential micronutrient for plants. Little is know about how iron is loaded in embryo during seed development. In this article we used Perls/DAB staining in order to reveal iron localization at the cellular and subcellular levels in different Brassicaceae seed species. In dry seeds of Brassica napus, Nasturtium officinale, Lepidium sativum, Camelina sativa, and Brassica oleracea iron localizes in vacuoles of cells surrounding provasculature in cotyledons and hypocotyl. Using B. napus and N. officinale as model plants we determined where iron localizes during seed development. Our results indicate that iron is not detectable by Perls/DAB staining in heart stage embryo cells. Interestingly, at torpedo development stage iron localizes in nuclei of different cells type, including integument, free cell endosperm and almost all embryo cells. Later, iron is detected in cytoplasmic structures in different embryo cell types. Our results indicate that iron accumulates in nuclei in specific stages of embryo maturation before to be localized in vacuoles of cells surrounding provasculature in mature seeds.

Angiotensin II AT1 receptors mediate neuronal sensitization and sustained blood pressure response induced by a single injection of amphetamine.

A single exposure to amphetamine induces neurochemical sensitization in striatal areas. The neuropeptide angiotensin II, through AT1 receptors (AT1-R) activation, is involved in these responses. However, amphetamine-induced alterations can be extended to extra-striatal areas involved in blood pressure control and their physiological outcomes. Our aim for the present study was to analyze the possible role for AT1-R in these events using a two-injection protocol and to further characterize the proposed AT1-R antagonism protocol. Central effect of orally administered AT1-R blocker (Candesartan, 3mg/kg p.o.×5days) in male Wistar rats was analyzed by spontaneous activity of neurons within locus coeruleus. In another group of animals pretreated with the AT1-R blocker or vehicle, sensitization was achieved by a single administration of amphetamine (5mg/kg i.p. - day 6) followed by a 3-week period off drug. On day 27, after receiving an amphetamine challenge (0.5mg/kg i.p.), we evaluated: (1) the sensitized c-Fos expression in locus coeruleus (LC), nucleus of the solitary tract (NTS), caudal ventrolateral medulla (A1) and central amygdala (CeAmy); and (2) the blood pressure response. AT1-R blockade decreased LC neurons' spontaneous firing rate. Moreover, sensitized c-Fos immunoreactivity in TH+neurons was found in LC and NTS; and both responses were blunted by the AT1-R blocker pretreatment. Meanwhile, no differences were found neither in CeAmy nor A1. Sensitized blood pressure response was observed as sustained changes in mean arterial pressure and was effectively prevented by AT1-R blockade. Our results extend AT1-R role in amphetamine-induced sensitization over noradrenergic nuclei and their cardiovascular output.

Model-based analysis supports interglacial refugia over long-dispersal events in the diversification of two South American cactus species.

Pilosocereus machrisii and P. aurisetus are cactus species within the P. aurisetus complex, a group of eight cacti that are restricted to rocky habitats within the Neotropical savannas of eastern South America. Previous studies have suggested that diversification within this complex was driven by distributional fragmentation, isolation leading to allopatric differentiation, and secondary contact among divergent lineages. These events have been associated with Quaternary climatic cycles, leading to the hypothesis that the xerophytic vegetation patches which presently harbor these populations operate as refugia during the current interglacial. However, owing to limitations of the standard phylogeographic approaches used in these studies, this hypothesis was not explicitly tested. Here we use Approximate Bayesian Computation to refine the previous inferences and test the role of different events in the diversification of two species within P. aurisetus group. We used molecular data from chloroplast DNA and simple sequence repeats loci of P. machrisii and P. aurisetus, the two species with broadest distribution in the complex, in order to test if the diversification in each species was driven mostly by vicariance or by long-dispersal events. We found that both species were affected primarily by vicariance, with a refuge model as the most likely scenario for P. aurisetus and a soft vicariance scenario most probable for P. machrisii. These results emphasize the importance of distributional fragmentation in these species, and add support to the hypothesis of long-term isolation in interglacial refugia previously proposed for the P. aurisetus species complex diversification.

Change in prevalence of congenital defects in children with Prader-Willi syndrome.

OBJECTIVE: The aim of this study was to assess the prevalence of congenital defects observed in patients with Prader-Willi syndrome (PWS) and to compare this prevalence with that described in the general population. In addition, these findings were correlated with the different etiologic subtypes. METHODS: A total of 180 children with PWS followed for 13 years were included in this study. Diagnosis was confirmed by the methylation test, and genetic subtypes were established by using fluorescence in situ hybridization or multiplex ligation-dependent probe amplification and microsatellite analyses. The prevalence of congenital defects was compared with national and international registries of congenital defects in the general population (Estudio Colaborativo Latinoamericano de Malformaciones Congénitas, European Surveillance of Congenital Anomalies, and the New York Registry). RESULTS: Twenty-two percent of the patients presented congenital defects with a risk of 5.4 to 18.7 times higher than that of the general population. The most frequent congenital defects were heart defects, renoureteral malformations, vertebral anomalies, hip dysplasia, clubfoot, and agenesis/hypoplasia of the corpus callosum. Each of these congenital defects was significantly more frequent in the children with PWS than in the general population. The congenital heart defects were more frequent in girls than in boys with PWS. No significant differences were found when the defects were correlated with the different etiologic subtypes. CONCLUSIONS: An increased prevalence of congenital defects was found in our PWS patients. This finding suggests the need for further studies in PWS children that allow physicians to detect the congenital defects found in this series and, thus, to anticipate complications, with the ultimate aim of enhancing the management of PWS patients.

Haga su diagnóstico / Make your diagnosis

El diagnóstico es Pitiriasis liquenoide crónica

Hippocampal effects of neuronostatin on memory, anxiety-like behavior and food intake in rats.

A 13-amino acid peptide named neuronostatin (NST) encoded in the somatostatin pro-hormone has been recently reported. It is produced throughout the body, particularly in brain areas that have significant actions over the metabolic and autonomic regulation. The present study was performed in order to elucidate the functional role of NST on memory, anxiety-like behavior and food intake and the hippocampal participation in these effects. When the peptide was intra-hippocampally administered at 3.0 nmol/µl, it impaired memory retention in both, object recognition and step-down test. Also, this dose blocked the hippocampal long-term potentiation (LTP) generation. When NST was intra-hippocampally administered at 0.3 nmol/µl and 3.0 nmol/µl, anxiolytic effects were observed. Also, the administration in the third ventricle at the higher dose (3.0 nmol/µl) induced similar effects, and both doses reduced food intake. The main result of the present study is the relevance of the hippocampal formation in the behavioral effects induced by NST, and these effects could be associated to a reduced hippocampal synaptic plasticity.

Different chronic cocaine administration protocols induce changes on dentate gyrus plasticity and hippocampal dependent behavior.

Hippocampus is a limbic structure that participates in learning and memory formation. Specifically the dentate gyrus has been described as a hippocampal subregion with high rates of plasticity and it is targeted by different psychoactive drugs modulating synaptic plasticity. Repeated cocaine administration induces sensitization to the locomotor effects and it is believed that sensitization involves the same mechanisms of drug seeking and relapse. Although, the mechanisms underlying sensitization is not fully understood. In this work we investigated the impact of repeated intraperitoneal administration of cocaine (15 or 20 mg/kg/day along 5 or 15 days respectively; and 15 mg/kg/day along 5 day followed by a challenge dose after three days of withdrawal) on the dentate gyrus synaptic plasticity, differentiating between sensitized and nonsensitized rats. Furthermore, we correlated changes on the hippocampal synaptic plasticity to memory retention. Our results revealed that the prevalence of cocaine sensitization (around 50%) was identical in all protocols used. The results found in the threshold to generate LTP were similar for all protocols used, being the threshold values cocaine-treated groups (sensitized and nonsensitized) significantly reduced compared to controls, observing the highest reduction in the sensitized group. Moreover, we observed a facilitated retention of recent memory formation only in sensitized animals the nonsensitized subjects remained at the control levels. In conclusion, sensitization to cocaine generates a high efficiency of hippocampal synaptic plasticity that may underlie the aberrant engagement of learning processes occurred during drug addiction.

The plastic phenomenon underlying the associative processes in the addictive properties of diazepam and other psychoactive drugs.

Benzodiazepines are commonly prescribed for the therapy of disorders such as anxiety and sleep disturbance, but develop dependence in many patients. In this review we discuss the impact of different brain areas that modulate the reward system in the development of tolerance and dependence to benzodiazepine and the associative processes underlying those fenomena.

Huperzia saururus, activity on synaptic transmission in the hippocampus.

Huperzia saururus (Lam.) Trevis. (Lycopodiaceae) known as cola de quirquincho is used in folk medicine to improve memory. The cholinergic neurons of the basal forebrain, including those in the medial septum, and in the vertical limbs of the diagonal band of Broca and the nucleus basalis of Meynert, provide a major source of cholinergic enervation of the cortex and hippocampus. These neurons have also been shown to play an important role in learning and memory processes. Thus, the effects of this traditional Argentinean species were studied in relation to its activity on synaptic transmission in the hippocampus. The alkaloid extract obtained first by decoction of the aerial parts and by subsequent alkaline extraction, was purified by using a Sephadex LH 20 packed column. Electrophysiological experiments were developed with the purified extract (E(2)) on rat hippocampus slices, thus eliciting long-term potentiation (LTP). Results show a marked increase in the hippocampal synaptic plasticity. The threshold value for generation of LTP was 22 +/- 1.01 Hz on average for E(2), while for controls it was 86 +/- 0.92 Hz. All of these factors could explain the use of Huperzia saururus as a memory improver as is reported in the ethnomedicine.

Hippocampus and locus coeruleus activity on rats chronically treated with diazepam.

The neural mechanisms underlying benzodiazepine (BZD) dependence remain equivocal. The present studies tested the hypothesis that similar neural circuitry might be involved in the effects of chronic 7-chloro-1-methyl-5-phenyl-3H-1,4-benzodiazepine-2(1H)-one, diazepam (DZ, Roche), administration and withdrawal. The results of our study showed an increased hippocampal synaptic plasticity in slices from rats chronically treated with DZ (5 mg/kg/18 days), assessed as a decrease of the threshold in the stimulation rate for long-term potentiation (LTP) elicitation. Rats with the same schedule of DZ administration but without signs of withdrawal behaved similarly to vehicle-treated ones (VEH), in the threshold to induce LTP. Furthermore, the activity of locus coeruleus (LC) norepinephrine (NE) neurons in rats tested 24 h after the last DZ injection showed a significant increase. On the other hand, rats that after chronic DZ administration did not develop signs of withdrawal and exhibited a similar pattern of discharge on LC-NE nucleus compared with their controls. We conclude that chronic DZ administration enhances both hippocampal synaptic plasticity and activity of LC-NE neurons. This neural system could be the biological substrate underlying the behavioral alterations accompanying chronic DZ administration and withdrawal.

Preexposure to drug administration context blocks the development of tolerance to sedative effects of diazepam.

The development of tolerance to benzodiazepine (BDZ) and other psychoactive agents such as morphine, alcohol, and barbiturates is thought to be a contingent or learning phenomenon. In a previous report, we demonstrated a positive correlation between the development of tolerance to the sedative effects of diazepam (DZ) and hippocampal synaptic plasticity. The results of the present work show that the development of tolerance to the hypolocomotor action of DZ (5 mg/kg) for 4 days and the associated increase in synaptic plasticity are context specific. Because animal preexposure to the drug administration context blocks both the tolerance sedative effects of DZ and the increased hippocampal synaptic plasticity, observed after 4 days of DZ administration, we propose the increased synaptic plasticity on hippocampal development as one of the biological substrates to the tolerance to DZ. Besides, the continuous administration of DZ did not induce a conditioning opponent response in these animals.

[Effects of fiber administration in the prevention of gallstones in obese patients on a reducing diet. A clinical trial]. / Efecto de la administración de fibra en la prevención de litiasis vesicular en obesos sometidos a dieta de reducción. Ensayo.

Nearly 30% of the obese patients treated with hypoenergetic diets for weight reduction develop gallstone disease (GD). Until the present time, the use of ursodeoxycholic acid (UDA) is the only available therapeutic measure to avoid the development of GD. Dietary fiber induce a bile acid synthesis. A double-blind clinical trial was conducted to compare the effect of rational diet plus UDA vs a rational diet supplemented with Psyllium plantago (Pp) for the prevention of GD in obese subjects undergoing a weight-reduction diet. Patients with a body mass index (BMI = weight in Kg/square height in m) of 30 Kg/m2 or more and with normal gallbladder and biliary tree ultrasound (GBUS) were included. Weight-reduction diets were individually calculated for each patient according to their energy expenditure (EE). Patients were randomly and blindly assigned either to group I (diet + 750 mg UDA + fiber placebo) or group II (diet + 15 g Pp+ UDA placebo). An anthropometric evaluation was performed to each patient before and after the two-month treatment, as well as resting EE by indirect calorimetry, GBUS and endoscopy for the determination of cholesterol crystals in duodenal bile. Weight reduction was similar in both groups (group I = 6 +/- 2 Kg vs group II = 6 +/- 3 Kg). GD development was observed in one patient of group I (5.5%) and two patients of group II (p > 0.05). All patients with GD lost a minimum of 4 Kg during the study period. GD development did not correlate with the presence of crystals in the duodenal bile at the beginning of the study. Our results suggest a beneficial effect of a rational diet with fiber supplementation to prevent GD development in obese patients included in a weight reduction program.

[Utilization of different microbiological markers in the study of Haemophilus influenzae]. / Utilización de diferentes marcadores microbiológicos en el estudio de Haemophilus influenzae.

The present study includes 178 Haemophilus influenzae strains isolated in different pediatric hospitals from Havana, Cuba, during 1991-1994, associated to divers infections (meningitis, respiratory sepsis, primary bacteremia). A combination of various typing and subtyping methods was used as epidemiological markers: serotyping (slide agglutination with diagnostical serum a-f and latex agglutination), biotyping according to Killian's procedures (by determination of indole production, urease and ornithine decarboxylase activity), subtyping by fermentative profiles according to Roberts' methods (glucose, maltose, xylose and fructose) and outer membrane protein profile subtyping (vesicles extraction by a modified Barenkamp's method, analysis by lineal and gradient SDS-PAGE and assessment according to our own classification system). Serotype b was identified in 89.3%, biotype I was the most frequent (79.1%), other biotypes (II, III, IV and V) were also identified. Fermentative profile D (glucose, maltose, xylose and fructose positive) was the most frequent (52.8%) while profile G (glucose, maltose, xylose positive and fructose negative) represented 20.2%. Other known profiles were present. PA2 (33.7%) was the most frequent OMP subtype. Even though 11 different protein subtypes were found, the 77.5% of the strains were located in only three OMP electrophoretic subtypes (PA2, PC1, LA2).

Resistencia antimicrobiana en Haemophilus influenzae en la ciudad de La Habana, Cuba / Antimicrobial resistance in Haemophilus influenzae in Habana City, Cuba

Se realizó un estudio de la resistencia a los antimicrobianos en 55 cepas de Haemophilus influenzae, aisladas en el período comprendido entre junio de 1992 y mayo de 1993 de igual número de pacientes pediátricos con infecciones invasivas y no invasivas; se utilizó el método de dilución de placas de agar, según las recomendaciones del NCCLS. De las cepas estudiadas, 49 por ciento, 47,3 por ciento y 27,3 por ciento fueron resistentes a la ampicilina, tetraciclina y cloranfenicol, respectivamente. No se detectó resistencia a la ceftriaxona, cefotaxima ni rifampicina. El 36,4 por ciento de las cepas fueron multiresistentes, presentándose una distribución heterogénea en 7 patrones de resistencia. Se demostró la producción de ß-lactamasa en 22 cepas (40 por ciento). La tasa de resistencia a las diferentes drogas fue mayor en cepas del serotipo b que en aquellas no b

Resistencia antimicrobiana en Haemophilus influenzae en la ciudad de La Habana, Cuba / Antimicrobial resistance in Haemophilus influenzae in Habana City, Cuba

Se realizó un estudio de la resistencia a los antimicrobianos en 55 cepas de Haemophilus influenzae, aisladas en el período comprendido entre junio de 1992 y mayo de 1993 de igual número de pacientes pediátricos con infecciones invasivas y no invasivas; se utilizó el método de dilución de placas de agar, según las recomendaciones del NCCLS. De las cepas estudiadas, 49 por ciento, 47,3 por ciento y 27,3 por ciento fueron resistentes a la ampicilina, tetraciclina y cloranfenicol, respectivamente. No se detectó resistencia a la ceftriaxona, cefotaxima ni rifampicina. El 36,4 por ciento de las cepas fueron multiresistentes, presentándose una distribución heterogénea en 7 patrones de resistencia. Se demostró la producción de ß-lactamasa en 22 cepas (40 por ciento). La tasa de resistencia a las diferentes drogas fue mayor en cepas del serotipo b que en aquellas no b (AU)

[Antimicrobial resistance in Haemophilus influenzae in the city of Havana, Cuba]. / Resistencia antimicrobiana en Haemophilus influenzae en la ciudad de La Habana, Cuba.

Fifty five Haemophilus influenzae strains were studied to determine their resistance to different antimicrobial drugs. They were isolated in Habana City, Cuba, during June 1992 to May 1993, from invasive and non invasive infections. An agar dilution method, according to NCCLS guidelines, was employed. We observed that 49%, 47.3% and 27.3% were resistant to ampicillin, tetracycline and chloramphenicol, respectively. beta-lactamase production was demonstrated in 22 strains (40%). There was neither resistance to ceftriaxone, cefotaxime nor rifampicin. 36.4% of the strains were multiresistant, being described 7 different resistance patterns. The rate of resistance to the drugs was substantially higher among serotype b than among non type b strains.