On-site extraction using a 3D printed device coated with Zn/Co-ZIF-derived carbon followed by an on-line SIA-HPLC-FL system for fluoroquinolones determination in wastewater.

A 3D printed device covered with Zn/Co-ZIF-derived carbon allows the on-site extraction of fluoroquinolones (FQs) from wastewater, avoiding the sample transportation to the laboratory, and the subsequent elution, separation and determination using an on-line flow system based on sequential injection analysis (SIA) coupled to HPLC-FL. Several parameters that affect the extraction efficiency and desorption were optimized including the sorption phase immobilization technique on the 3D device, extraction time, pH effect, sample volume as well as the type of eluent, eluent volume, and flow rate. Under optimum conditions, detection limits of 3-9 ng L-1 were achieved for norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin and difloxacin. The precision expressed as relative standard deviation (%RSD, n = 3), showed intraday and interday ranges of 1.5-5.3% and 2.8-5.7%, respectively, demonstrating a good precision of the proposed methodology. To assess matrix effects and accuracy of the proposed method in real samples, recovery studies were performed without and with FQs spiked at different concentrations (0.5-10 µg L-1) to wastewater samples, showing good recoveries in the range of 91-104%. The results allow to confirm the applicability of MOF-derived carbons as adsorbents for on-site extraction, and the satisfactory separation and quantification of FQs by a SIA-HPLC-FL on-line system after their desorption with small eluent volumes.

Pregnancy and birth rate outcomes in alpacas (Vicugna pacos) inseminated with frozen semen using two commercial extenders.

The aim of this study was to evaluate alpaca pregnancy outcomes and birth rates of females inseminated with frozen semen using two commercial extenders. A total of 18 ejaculates from 8 adult alpaca males were obtained with artificial vagina, and macroscopic and microscopic semen characteristics were assessed. Afterwards, samples were divided into two aliquots, diluted with Biladyl® B or AndroMed®, and cooled for 2 h at 5°C. At that moment, sperm motility was evaluated, and samples were frozen through a gradual descent of temperature using a liquid nitrogen tank. To analyse frozen sperm quality, samples were thawed at 38°C for 30 s. Even though a significant decrease in sperm motility and viability was detected when thawed (p < .05), no superiority was found between the two commercial extenders (Biladyl® B vs. AndroMed®). A total of 36 alpaca females were artificially inseminated (AI) between 30 and 34 h post-injection of a GnRH analogue, administered when a growing dominant follicle was detected through transrectal palpation and ultrasonography. Obtained pregnancy rates were similar between Biladyl® B (33.3%, 6/18) and AndroMed® (22.2%, 4/18). No significant differences were detected in birth rates between the two tested extenders, obtaining 4 and 3 births for Biladyl® and AndroMed®, respectively. In conclusion, alpaca pregnancies and alive offspring can be obtained through AI with frozen semen at similar efficiency rates using commercial diluents, Biladyl® B or AndroMed®.

Functional identification of a Streptomyces lividans FKBP-like protein involved in the folding of overproduced secreted proteins.

Some bacterial peptidyl-prolyl cis/trans isomerases (PPIases) are involved in secretory protein folding after the translocation step. Streptomyces lividans has been used as a host for engineering extracellular overproduction of homologous and heterologous proteins in industrial applications. Although the mechanisms governing the major secretory pathway (Sec route) and the minor secretory pathway (Tat route) are reasonably well described, the function of proteins responsible for the extracellular secretory protein folding is not characterized as yet. We have characterized a Tat-dependent S. lividans FK506-binding protein-like lipoprotein (FKBP) that has PPIase activity. A mutant in the sli-fkbp gene induces a secretion stress response and affects secretion and activity of the Sec-dependent protein α-amylase. Additionally, propagation in high copy number of the sli-fkbp gene has a positive effect on the activity of both the overproduced α-amylase and the overproduced Tat-dependent agarase, both containing proline cis isomers. Targeted proteomic analyses showed that a relevant group of secreted proteins in S. lividans TK21 are affected by Sli-FKBP, revealing a wide substrate range. The results obtained indicate that, regardless of the secretory route used by proteins in S. lividans, adjusting the expression of sli-fkbp may facilitate folding of dependent proteins when engineering Streptomyces strains for the overproduction of homologous or heterologous secretory proteins.

Comparative genomics to bridge Vicia faba with model and closely-related legume species: stability of QTLs for flowering and yield-related traits.

This study presents the development of an enhanced map in faba bean. The map contains 258 loci, mostly gene-based markers, organized in 16 linkage groups that expand 1,875 cM, with an average inter-marker distance of 7.26 cM. The combination of EST-derived markers with a number of markers physically located or previously ascribed to chromosomes by trisomic segregation, allowed the allocation of eight linkage groups (229 markers), to specific chromosomes. Moreover, this approach provided anchor points to establish a global homology among the faba bean chromosomes and those of closely-related legumes species. The map was used to identify and validate, for the first time, QTLs controlling five flowering and reproductive traits: days to flowering, flowering length, pod length, number of seeds per pod and number of ovules per pod. Twelve QTLs stable in the 2 years of evaluation were identified in chromosomes II, V and VI. Comparative mapping suggested the conservation of one of the faba bean genomic regions controlling the character days to flowering in other five legume species (Medicago, Lotus, pea, lupine, chickpea). Additional syntenic co-localizations of QTLs controlling pod length and number of seeds per pod between faba bean and Lotus japonicus are likely. The new genetic map opens the way for further translational studies between faba bean and related legume species, and provides an efficient tool for breeding applications such as QTL analysis and marker-assisted selection.

Depressive-type emotional response pattern in impulsive-aggressive patients with borderline personality disorder.

INTRODUCTION: Borderline personality disorder (BPD) is typically characterized by severe affective dysregulation leading to impulsive behaviors. Accordingly, preliminary data suggest the hypothesis that BPD patients could have a specific and altered pattern of subjective emotional response to stimuli. The nature of the emotional response in BPD can be compared with other affective disorders and provide further insight on the nosological proximity with other psychiatric disorders. METHODS: Subjective emotional response was investigated in 19 patients with DSM-IV BPD with no current depressive episode and in 19 healthy control subjects by using the International Affective Picture System (IAPS). The intensity of arousal, valence and dominance was rated in response to 60 images categorized as pleasant, unpleasant and neutral by using a self-assessment instrument. ANOVA of multiple factors was used for between-groups comparisons. RESULTS: The obtained pattern showed that BPD patients considered the unpleasant and neutral images as less aversive than controls, but the activation that these images induced was higher. Patients showed significantly greater arousal than controls for unpleasant and neutral images (p<0.05) but presented greater valence (more positive emotion) for these images (p<0.05). In addition, BPD patients showed lower dominance (greater insecurity and dyscomfort) for positive images (p<0.05). CONCLUSIONS: The subjective emotional response pattern of BPD patients suggests a trait of vulnerability to pleasant stimuli and is similar to the pattern found in depressive patients in previous studies. This supports the evidence that BPD could in part be related with the spectrum of the affective temperament and affective disorders.

Identification and characterization of NBS-LRR class resistance gene analogs in faba bean (Vicia faba L.) and chickpea (Cicer arietinum L.).

A PCR approach with degenerate primers designed from conserved NBS-LRR (nucleotide binding site-leucine-rich repeat) regions of known disease-resistance (R) genes was used to amplify and clone homologous sequences from 5 faba bean (Vicia faba) lines and 2 chickpea (Cicer arietinum) accessions. Sixty-nine sequenced clones showed homologies to various R genes deposited in the GenBank database. The presence of internal kinase-2 and kinase-3a motifs in all the sequences isolated confirm that these clones correspond to NBS-containing genes. Using an amino-acid sequence identity of 70% as a threshold value, the clones were grouped into 10 classes of resistance-gene analogs (RGA01 to RGA10). The number of clones per class varied from 1 to 30. RGA classes 1, 6, 8, and 9 were comprised solely of clones isolated from faba bean, whereas classes 2, 3, 4, 5, and 7 included only chickpea clones. RGA10, showing a within-class identity of 99%, was the only class consisting of both faba bean and chickpea clones. A phylogenetic tree, based on the deduced amino-acid sequences of 12 representative clones from the 10 RGA classes and the NBS domains of 6 known R genes (I2 and Prf from tomato, RPP13 from Arabidopsis, Gro1-4 from potato, N from tobacco, L6 from flax), clearly indicated the separation between TIR (Toll/interleukin-1 receptor homology: Gro1-4, L6, N, RGA05 to RGA10)- and non-TIR (I2, Prf, RPP13, RGA01 to RGA04)-type NBS-LRR sequences. The development of suitable polymorphic markers based on cloned RGA sequences to be used in genetic mapping will facilitate the assessment of their potential linkage relationships with disease-resistance genes in faba bean and chickpea. This work is the first to report on faba bean RGAs.

Glucose sensing through the Hxk2-dependent signalling pathway.

In this work, we describe the hexokinase 2 (Hxk2) signalling pathway within the yeast cell. Hxk2 and Mig1 are the two major factors of glucose repression in Saccharomyces cerevisiae. The functions of both proteins have been extensively studied but there is no information about possible interactions among them in the repression pathway. Our results demonstrate that Hxk2 interacts directly with Mig1 in vivo and in vitro and that the ten amino acids motif between K6 and M15 is required for their interaction. This interaction has been detected at the DNA level both in vivo by chromatin immunoprecipitation experiments and in vitro using purified proteins and a DNA fragment containing the MIG1 site of the SUC2 promoter. This demonstrates that the interaction is of physiological relevance. Our findings show that the main role of Hxk2 in the glucose signalling pathway is the interaction with Mig1 to generate a repressor complex located in the nucleus of S. cerevisiae.

Population dose in the vicinity of old Spanish uranium mines.

Regional surveys were conducted to determine exposure to natural sources of radiation for people in the vicinity of old Spanish uranium mines. The surveys evaluated indoor radon concentrations and outdoor and indoor external gamma dose rates. Indoor radon concentrations were measured in 222 dwellings by means of nuclear track-etched detectors. The terrestrial gamma ray dose rate was measured outdoors and indoors at a total of 256 points and 115 points, respectively. Estimates mean annual effective doses for the six areas studied ranged from 3.2 to 5.1 mSv per year, which is between 1.2 and 2 times higher than the average national value.

Natural radiation exposure in the vicinity of Spanish nuclear power stations.

Nationwide and regional surveys have been conducted to evaluate natural radiation exposure of people in the vicinity of the Spanish nuclear power stations. In these surveys, indoor radon, external gamma dose rates outdoors and indoors and radioactivity in soils were measured. Measurements of indoor radon concentrations were performed in 214 dwellings by using nuclear track-etched detectors. External exposure from terrestrial gamma rays outdoors and indoors were measured in a total of 596 points. Radioactivity in soil samples collected at 293 sampling sites are also reported in this paper. By taking into account all sources of natural radiation exposure, mean annual effective doses to the population in the surroundings of the six Spanish nuclear power stations have been estimated and compared with those coming from their normal operation.

Eyaculación precoz: utilidad del tiempo-al-orgasmo como criterio diagnóstico / Early ejaculatión: time to orgasm utility as diagnosis criteria

Aunque no existe un criterio cuantitativo de tiempo de la definición de Eyaculación Precoz (EP) del DSM-IV, sería útil disponer de un parámetro objetivo referente al tiempo más común en alcanzar el orgasmo, a fin de orientar el diagnóstico así como las expectativas del paciente que padece de eyaculación rápida. Nuestro propósito es el de establecer este punto de referencia. 56 pacientes masculinos que consultaron por disfunción sexual (20por disfunción eréctil-DE-sola, 28 por EP y 8 por DE+EP, sumado así 36 pacientes refiriendo EP) respondieron relativo a aspectos de su eyaculación y orgasmo. comparamos luego los tiempos-al-orgasmo reportados por los pacientes con EP con los mismos tiempos en el grupo de no EP. Elaboramos entonces un gráfico de distribución utilizando un cuadro de doble entrada para determinar la sensibilidad y especificidad de cada párametro de tiempo reportado a fin de evaluar la EP. Los tiempos extremos de cada grupo fueron excluidos. El tiempo-al-orgasmopromedio en el grupo de EP fue de 1,44 minutos vs. 5,18 minutos en el grupo de no EP. Un punto de corte de 3 minutos nos da la mejor combinación de sensibilidad y especificidad diagnóstica para nuestro grupo, siendo de 94 por ciento y 47 por ciento, respectivamente. Aunque el tiempo-al-orgasmo no es un parámetro considerado por el DSM-IV para la definición de EP, existe una diferencia obvia en el tiempo promedio mostrado en ambos grupos. Un punto de corte de 3 minutos da una buena referencia para distinguir y orientar al paciente y al médico acerca de lo que es más común.

[Errors in the oral administration of medicines in two health centres] / Errores en la administración de medicamentos por vía oral en dos centros sociosanitarios.

OBJECTIVE: To know how medicines should be administered, to detect if there are any interaction whith food and the most frequent errors. METHOD: A descriptive study has been carried out in two health centres where we have evaluated if the oral medicines have been correctly dispensed. So as to do this, the summary of medicines' characteristics and/or medicine prospects were used. RESULTS: The daily administered medication for 40 patients was evaluated during one day in two different hospitals. We administered 274 medicines in hospital A and 238 medicines in hospital B wich correspond to 344 and 330 oral dosis. In hospital A 138 (23,4%) mistakes were made and another 65 potential errors were detected. It is unsure if these potential errors were made or not as we do not know if the medication was taken correctly by the patient. In hospital B 95 (25,0%) errors were committed. No other possible errors were observed. The most frequent errors that have been commited are: not to dilute the medication with water, to administer the medicine with milk and/or antiacid medicine, to take medicine with meals instead of taking them in abstinence. Not warning the pacient to swallow medicines without chewing them. Other errors were concerning the relationship between medication and meals. DISCUSSION: Many of the mistakes that are made when taking medicines are due to the lack of knowledge of the correct usage or lack of information.

[Typhoid fever in school children: by what measures is the modification of the clinical course due to oral vaccination?]. / Fiebre tifoidea en escolares: en que medida la clinica es modificada por el uso de una vacuna oral?

The clinical course of infection by Salmonellae was compared between patients who had been vaccinated against typhoid fever using the Ty21a vaccine and those who had not. Of 2566 bacteriological confirmed cases 84% were infected with S typhi, 14% with S paratyphi B and 2% with S paratyphi A. Among patients with typhoid fever, 34% were treated in hospital, 3.5% had relapses, 5.4% developed complications and 1 patient died (0.05%). Among patients with paratyphoid fever, 18% were treated in hospital, 0.6% had relapses, 1.4% developed complications and there were no deaths. These figures were similar among vaccinated and non-vaccinated cases. A slightly greater proportion of vaccinated cases were treated in hospital (38 vs 30%). Thus, use of oral vaccination against typhoid fever does not alter the clinical course of infection with Salmonellae.

Acid-base disturbances in cardiogenic pulmonary edema.

Eighty-one consecutive cases of uncomplicated cardiogenic pulmonary edema (CPE) were retrospectively graded for severity of chest roentgenogram (CXR) changes and grouped according to primary acid-base abnormalities, either single or mixed. Mean age was 72, 50 male, 31 female. Twenty-three percent had no acid-base disturbances (ABD). Isolated respiratory alkalosis was most common (41%), followed by metabolic acidosis, 22%; metabolic alkalosis, 10%, and respiratory acidosis, 9%. Age, sex, race distribution, morbidity and mortality were not significantly different between the groups. Overall mortality was 17%. Significantly higher mortality was associated with age over 70, pH less than 7.4, and presence of acute myocardial infarction. CXR scores did not correlate with pH, pCO2 or pO2, mortality or morbidity. Some patients with the most severe ABDs recovered while others, who had no ABD on presentation, eventually died. Thus, in 81 consecutive episodes of uncomplicated CPE, isolated respiratory alkalosis was the commonest ABD, occurring in 41%. No correlation was found between ABD and severity of CPE, morbidity or mortality.

Chronic effects of nitrendipine on renal hemodynamics and tubular transport.

The effects of nitrendipine (10 mg, during acute clearance experiments) given both acutely and after 2 weeks of administration were examined in normal and hypertensive subjects. At the initiation of therapy, nitrendipine caused an increase in glomerular filtration rate and effective renal plasma flow in the hypertensive, but not in the normal, group. Percentage excretion rates of sodium (%ENa) and phosphate (%EPi) and free water clearance (CW) increased in both groups at the initiation of therapy. After 2 weeks of nitrendipine therapy repeat acute clearance studies showed that the drug no longer increased glomerular filtration rate or effective renal plasma flow in hypertensive subjects, the increases in %ENa and CW persisted in normal subjects and trended upward in hypertensive subjects, and the increase in %EPi persisted in both normal and hypertensive individuals. We conclude that nitrendipine is not sodium retentive after chronic therapy and the acute increase in %ENa, %EPi, and CW after its initial administration suggests a proximal tubular effect.

Antibodies to porin antigens of Salmonella typhi induced during typhoid infection in humans.

Immunoglobulin G (IgG)- and IgM-specific antibody titers against Salmonella typhi Ty2 porins have been measured in 30 paired typhoid sera by enzyme-linked immunosorbent assay. These studies have found that IgG serum titers of acute and convalescent sera were 625 and 5,000 times higher, respectively than the control serum titers. The same typhoid sera were titrated with S. typhi Ty2 flagellin and S. typhi lipopolysaccharide. The titers against these antigens were considerably lower than those against the porins. The highest IgM-specific titer has also been found against porins in convalescent-phase sera. However, the largest increase in IgM-specific titer compared with the control group titer was obtained against flagellin during the acute phase of typhoid. The lowest increases in antibody titer were obtained with the IgM-specific anti-lipopolysaccharide in both types of sera. This may be because many normal individuals in endemic areas already have IgM titers against lipopolysaccharide. This study has provided good evidence that porins are excellent antigens and that IgG-specific antiporin titers may be of diagnostic value in typhoid infections in endemic areas.

Detection of urinary Vi antigen as a diagnostic test for typhoid fever.

Since Vi antigen is limited primarily to Salmonella typhi, it has been thought that detection of the antigen may be a useful method for diagnosing acute typhoid fever. The slide coagglutination method and enzyme-linked immunosorbent assay have recently been suggested as ways to detect small quantities of Vi antigen in urine. In Santiago, Chile, we compared the results of these two methods in patients with acute typhoid fever, paratyphoid fever, and other febrile illnesses and in afebrile control subjects. Using a cut-off value that maximally separated typhoid patients from controls, the enzyme-linked immunosorbent assay was positive in 62.4% of 141 patients with culture-proven typhoid infections and in 13.2% of 159 afebrile control subjects. The enzyme-linked immunosorbent assay was false positive in 64.7% of 34 culture-proven paratyphoid A or B patients and 47.1% of 21 patients with other nontyphoidal febrile illnesses. The coagglutination test was positive in 34% of typhoid patients, 14% of afebrile control subjects, and 46% of febrile control subjects. We conclude that these tests when performed with the Vi antibodies employed in this study are of little value for the diagnosis of typhoid fever in this setting.