Glucose transporter GLUT8 mRNA expression in intestinal Caco-2 cells is regulated by growth and metabolism.
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Caco-2 Cells↗
Biomedical subjects
A Romero
Publications and source records attributed to A Romero.
Copeptin, derived from isotocin precursor, is a probable prolactin releasing factor in carp.
Control of prolactin (PRL) release is of crucial importance for the multiple functions exerted by PRL in vertebrates. Recently identified hypothalamic PRL-releasing peptides displayed additional neuromodulatory activities and in fish only few could be detected close to lactotrophs. Here we describe the C-terminal peptide processed from the carp isotocin precursor as probable physiologically relevant regulator of PRL release in carp. The amino acid sequence derived from the complete isotocin precursor gene of Cyprinus carpio, predicted a C-terminal peptide uncleaved between the neurophysin (Np) and copeptin (Cp) domain. Accordingly, antibodies against synthetic Np- and Cp-specific oligopeptides both immunodetected a 13kDa protein (cNpCp) in total pituitary proteins and showed abundant immunoreaction in hypothalamic axons in direct contact with lactotrophs in the rostral pars distalis of carp pituitary gland sections. Finally, incubation of cultured carp pituitary explants with purified carp cNpCp resulted in a potent stimulation of PRL release.
Amino Acid Sequence↗
Modular changes of cis-regulatory elements from two functional Pit1 genes in the duplicated genome of Cyprinus carpio.
The pituitary-specific transcription factor Pit1 is involved in its own regulation and in a network of transcriptional regulation of hypothalamo-hypophyseal factors including prolactin (PRL) and growth hormone (GH). In the ectotherm teleost Cyprinus carpio, Pit1 plays an important role in regulation of the adaptive response to seasonal environmental changes. Two Pit1 genes exist in carp, a tetraploid vertebrate and transcripts of both genes were detected by RT-PCR analysis. Powerful comparative analyses of the 5'-flanking regions revealed copy specific changes comprising modular functional units in the naturally evolved promoters. These include the precise replacement of four nucleotides around the transcription start site embedded in completely conserved regions extending upstream of the TATA-box, an additional transcription factor binding site in the 5'-UTR of gene-I and, instead, duplication of a 9 bp element in gene-II. Binding of nuclear factors was assessed by electro mobility shift assays using extracts from rat pituitary cells and carp pituitary. Binding was confirmed at one conserved Pit1, one conserved CREB and one consensus MTF1. Interestingly, two functional Pit1 sites and one putative MTF1 binding site are unique to the Pit1 gene-I. In situ hybridization experiments revealed that the expression of gene-I in winter carp was significantly stronger than that of gene-II. Our data suggest that the specific control elements identified in the proximal regulatory region are physiologically relevant for the function of the duplicated Pit1 genes in carp and highlight modular changes in the architecture of two Pit1 genes that evolved for at least 12 MYA in the same organism.
Animals↗
Comparative study of disc diffusion and microdilution methods in susceptibility testing of micafungin against Candida species.
OBJECTIVES: To compare the Clinical Laboratory Standards Institute CLSI M44-A disc diffusion (DD) and M27-A2 broth microdilution (MD) methods for determining the susceptibility of Candida spp. to micafungin (FK463). PATIENTS AND METHODS: A total of 355 clinical yeast isolates including 270 Candida albicans, 45 Candida glabrata, 24 Candida krusei, 11 Candida tropicalis and 5 Candida parapsilosis were studied. The MIC of micafungin was determined by following the CLSI M27-A2 guidelines (MD). Endpoints were defined as the lowest concentration of micafungin resulting in partial inhibition (IC(50)) of visual growth after 24/48 h of incubation at 35 degrees C. Final concentrations were 0.008-4 mg/L of micafungin. DD testing was performed using a CLSI M44-A document with 2.5 mug micafungin discs. Zone diameter endpoints were read after 24/48 h of incubation at 35 degrees C. Arbitrary breakpoints were 4 mg/L for MD and 15 mm for DD. RESULTS: The best correlation was observed when we read MD 48 h/DD 24 and 48 h (97%). When the reading was MD 24 h/DD 24 and 48 h the percentage of correlation was 95.2%. CONCLUSIONS: The DD method performs well for testing the susceptibility of Candida spp. to micafungin. More studies involving more Candida strains with elevated MIC values are needed.
AIDS-Related Opportunistic Infections↗
Cyclic fatty acid monomer formation in domestic frying of frozen foods in sunflower oil and high oleic acid sunflower oil without oil replenishment.
During the frying process, oxidation, hydrolysis, polymerization, isomerization, and cyclization occur. Polymers and Cyclic fatty acid monomers (CFAM) are potentially toxic, and the latter are detected at relatively low levels (0.01-0.7%) in used frying oils. Twenty fryings of different frozen foods were carried out over 10 consecutive days in sunflower oil (SO) and in high oleic acid sunflower oil (HOSO). Fatty acid methyl ester derivates were hydrogenated with platinum oxide catalyst under hydrogen. Ethyl palmitate was added as an internal standard before hydrogenation. The CFAM obtained were isolated, concentrated and quantified by HPLC using a reverse-phase column followed by gas chromatography. Linear adjustments between total and individual CFAM content and the number of frying operations performed with both oils were established by analysis of variance. The comparison between linear equation adjustments of both oils was performed by a two-way analysis of covariance. After 20 fryings 15.4 +/- 0.06 g polar content/100 g oil, 7.15 +/- 0.08 g polymers/ oil, 11.52 +/- 0.08 g polymers/100g oil and 855 +/- 8.9 mg CFAM/kg oil were detected in SO. A 10 mg/100 mg oil of altered fatty acid content correspond to 700 mg/kg CFAM, while 25% polar material and 10% polymer content would correspond to about 850-1,000 mg CFAM/kg oil. Data suggest that frying with SO produces in each new frying 9 mg CFAM/kg more than frying with HOSO (p < 0.001). After frying cyclopentyl structures were more than twice as abundant as cyclohexyl fatty acids in both oils. Bicyclic compound formation was significantly higher in SO (p < 0.001). Because digestion and absorption of polar material, polymers and CFAM occur, data clearly show the advantageousness and advisability of frying with HOSO rather than SO.
Cooking↗
Cloning of the prepro C-RFa gene and brain localization of the active peptide in Salmo salar.
In all vertebrates, the synthesis and release of prolactin (Prl) from pituitary lactotroph cells is tightly controlled by hypothalamic factors. We have cloned and characterized a hypothalamic cDNA from Atlantic salmon (Salmo salar) encoding C-RFa, a peptide structurally related to mammalian Prl-releasing peptide (PrRP). The deduced preprohormone precursor is composed of 155 amino acid residues presenting a 87.1% similarity to chum salmon C-RFa and a 100% similarity to all fish C-RFa in the bioactive precursor motifs. C-RFa-immunoreactive perikarya and fibres were located in the brain of S. salar, especially in the hypothalamus, olfactory tract, optic tectum and cerebellum. In contrast, immunolabelled fibres were not observed in the pituitary stalk or in the hypophysis. However, interestingly, we detected immunolabelled cells in the rostral pars distalis of the pituitary in the basolateral region in which Prl is synthesized. These results were confirmed by obtaining a strong signal by using reverse transcription/polymerase chain reaction (RT-PCR) on mRNA from both hypothalamus and pituitary. These data show, for the first time, by immunohistochemistry and RT-PCR, that C-RFa is produced in pituitary cells. Finally, based on these results, a possible function for C-RFa as a locally produced PrRP in this teleost is discussed.
Amino Acid Sequence↗
Association of polymorphisms in the bovine FASN gene with milk-fat content.
Fatty acid synthase (FASN) is a multifunctional protein that carries out the synthesis of fatty acids so it plays a central role in de novo lipogenesis in mammals. Previously, we defined the genetic structure and expression of the bovine FASN gene. Our mapping studies placed FASN on BTA19 (19q22) where several quantitative trait loci (QTL) affecting milk-fat content and related traits have been described. This study was conducted to identify polymorphisms in the bovine FASN gene and to study their association with milk-fat content. The bovine FASN gene was screened for polymorphisms in two cattle breeds. Sequence analysis revealed several single nucleotide polymorphisms (SNPs), and two of them were analysed: a G>C substitution in the untranslated exon 1 (g.763G>C), altering a potential Sp1 transcription factor-binding site, and an A>G substitution in exon 34 (g.16009A>G), which determines a non-conservative substitution of threonine by alanine. Allele-specific amplification of the SNPs in FASN revealed significant frequency differences for both polymorphisms in Holsteins with high and low breeding values for milk-fat content. The intragenic haplotypes comprising exon 1 (alleles G and C) and exon 34 (alleles A and G) polymorphisms were studied, and the existence of linkage disequilibrium between these SNPs was found (D(CG) = 0.048, P < 0.001). Our results suggest that the FASN gene polymorphisms contribute to variation in milk-fat content. We propose that the bovine FASN gene is a candidate gene for a milk-fat content QTL.
Alleles↗
Role of bacteria in the oviposition behaviour and larval development of stable flies.
Stable flies, Stomoxys calcitrans (L.), are the most important pests of cattle in the United States. However, adequate management strategies for stable flies, especially for pastured cattle, are lacking. Microbial/symbiont-based approaches offer novel venues for management of insect pests and/or vector-borne human and animal pathogens. Unfortunately, the fundamental knowledge of stable fly-microbial associations and their effect on stable fly biology is lacking. In this study, stable flies laid greater numbers of eggs on a substrate with an active microbial community (> 95% of total eggs oviposited) than on a sterilized substrate. In addition, stable fly larvae could not develop in a sterilized natural or artificial substrate/medium. Bacteria were isolated and identified from a natural stable fly oviposition/developmental habitat and their individual effect on stable fly oviposition response and larval development was evaluated in laboratory bioassays. Of nine bacterial strains evaluated in the oviposition bioassays, Citrobacter freundii stimulated oviposition to the greatest extent. C. freundii also sustained stable fly development, but to a lesser degree than Serratia fanticola. Serratia marcescens and Aeromonas spp. neither stimulated oviposition nor supported stable fly development. These results demonstrate a stable fly bacterial symbiosis; stable fly larval development depends on a live microbial community in the natural habitat, and stable fly females are capable of selecting an oviposition site based on the microbially derived stimuli that indicate the suitability of the substrate for larval development. This study shows a promising starting point for exploiting stable fly-bacterial associations for development of novel approaches for stable fly management.
Analysis of Variance↗
Structural and functional implications of the hexokinase-nickel interaction.
The interaction between nickel and yeast hexokinase was studied. The binding of nickel showed a positive cooperativity, and saturation was not reached. The nickel binding induced modifications in the secondary structure of the protein; thus, a lost of alpha helix and beta turns, as well as an increase of the random structure and beta sheet was observed. The monomer/dimmer equilibrium of the protein was modified in the presence of nickel, and the monomer state was mainly obtained at the highest nickel concentrations studied. These changes on the protein structure caused a decrease in the enzyme activity. According to kinetic studies, nickel caused a non-competitive inhibition when glucose was the variable substrate and a linear competitive inhibition when ATP was the variable substrate.
Adenosine Triphosphate↗
Toxic effects of cadmium on the regulatory mechanism of dopamine and serotonin on prolactin secretion in adult male rats.
This work was designed to analyze the effects of cadmium on the regulatory mechanism of prolactin in cadmium-exposed rats. Adult male rats were given cadmium at a dose of 25 ppm of cadmium chloride (CdCl2) in the drinking water for 1 month. At the end of the treatment, the rats were killed at six different time intervals throughout a 24 h cycle to measure circulating prolactin levels and dopamine and serotonin content in the median eminence and in both anterior and posterior pituitary. Control and cadmium-exposed animals exhibited significant time of day-dependent variations in plasma prolactin levels and in dopamine and serotonin concentration in all analyzed tissue. Cadmium exposure did not modify the mean values of the hormone around the clock but it modified the amplitude of the secretory peaks at 08:00 and 12:00 h. Cadmium decreased dopamine content in the median eminence, while increased its content in the posterior pituitary and induced a phase advanced peak at 20:00 h. On the other hand, serotonin content was not modified in the median eminence. Only differences in specific time points were observed, while serotonin concentration in anterior and posterior pituitary were increased in cadmium-treated rats. Differences in time peaks were also observed. The negative correlation between plasma prolactin and dopamine content in pituitary, and between plasma levels of the hormone and serotonin content in posterior pituitary, disappeared in cadmium-treated animals.
Animals↗
Toxic effects of cadmium on GABA and taurine content in different brain areas of adult male rats.
This work assesses the possible changes in gamma amino butyric acid (GABA) and taurine content in the hypothalamus, the median eminence and striatum after the exposure to various doses of cadmium. Cadmium chloride (CdCl2) was administered in the drinking water at the doses of 5, 10, 25, 50 or 100 ppm to adult male rats for 1 month. In the anterior hypothalamus, taurine and GABA content decreased with the dose of 10 ppm of CdCl2 only. Cadmium exposure decreased both GABA and taurine content in mediobasal hypothalamus except for the 50 ppm dose. In posterior hypothalamus GABA and taurine content was not affected by cadmium treatment. As far as the median eminence, 5 or 10 ppm of CdCl2 increased taurine concentration, and at a dose of 5 ppm enhanced GABA content. A significant decrease of GABA and taurine concentration was seen in the striatum at any dose of cadmium used. The concentration of cadmium increased in the hypothalamus and in the striatum in animals receiving CdCl2 in the drinking water at doses of 25, 50 or 100 ppm. The results indicate that cadmium globally decreased GABA and taurine content in the brain areas studied through effects that were not dose dependent.
Animals↗
Utility of point-of-care haemoglobin measurement in the HemoCue-B haemoglobin for the initial diagnosis of anaemia.
Measurement of haemoglobin (Hb) concentration provides a reliable indication of the presence and severity of anaemia. However, other laboratory parameters are usually requested as well, leading to an increase in socio-sanitary costs. Accordingly, this study was undertaken to ascertain the reliability of point-of-care Hb determination with the portable photometer HemoCue-B haemoglobin (HBH) and to evaluate its utility for the initial diagnosis of anaemia. Hb was measured (x3) in 20 venous blood samples diluted with saline (v/v; 1 : 0, 2 : 1, 1 : 1, 2 : 1 and 3 : 1) to obtain a wide range of Hb and in venous and capillary blood samples from 247 primary health care patients. All HBH results were compared with those yielded by the reference cell counter Pentra 120 Retic (ABX). In diluted samples, Hb values obtained with either method were not significantly different (ABX-HBH, -0.01 +/- 0.32 g/dl; 95% CI, -0.04 to 0.028 g/dl) and showed an excellent Pearson's coefficient of correlation (r = 0.992; P < 0.01). HBH provides accurate values if at least 4 mul of blood is loaded into the cuvette. There were no significant differences between Hb measured in venous (v) and capillary (c) blood samples in primary care patients. Eighteen anaemic patients were detected by ABX measurements (7.3%; 15 female/3 male), 18 by HBHv (specificity, 100%; sensitivity, 100%) and 25 by HBHc (eight false positives; one false negative; specificity, 94.4%; sensitivity, 96.5%). Compared with ABX, HBH provides accurate and precise measurements for a wide range of Hb and its use in primary health care seems to be a good method for the initial diagnosis of anaemia.
Anemia↗
[Histoplasmosis in a renal transplant patient].
The case of a Spanish kidney transplant patient who developed disseminated histoplasmosis approximately one year and a half after transplantation without having previously visited or travelled to endemic areas of histoplasmosis is presented. To our knowledge this is the first case of this disease in a kidney transplant patient in Spain without epidemiologic antecedent. The study of anti-histoplasm antibodies by complement fixation of the donor and recipient did not safely clarify the mechanism of contagion.
Histoplasmosis↗
Cadmium exposure differentially modifies the circadian patterns of norepinephrine at the median eminence and plasma LH, FSH and testosterone levels.
This work was designed to analyze the cadmium effects on time-of-day variations of norepinephrine (NE) content in median eminence and on plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone in adult male rats. Rats were given cadmium at a dose of 25 ppm of cadmium chloride (CdCl(2)) in the drinking water for 1 month. Significant 24-h changes of NE content in the median eminence, plasma LH and testosterone levels occurred in control animals. Cadmium exposure induced a phase advance of the nocturnal peak of NE content that was described in the control group, to 12h and increased its amplitude. However, the mean NE content was not changed by cadmium. Metal exposure abolished the daily pattern of plasma LH levels, although the mean levels of the hormone were not modified by cadmium. For testosterone, the metal increased the amplitude of its nocturnal peak and induced the appearance of another peak during the light phase at 12h, thus increasing the mean plasma levels of this hormone. An interaction between the metal and time for NE and plasma testosterone levels was observed. These data suggest that cadmium exerts differential effects at the median eminence, the pituitary and the testes, that may explain the changes in the 24-h pattern of plasma testosterone levels.
Animals↗
Documentation of subtype C HIV Type 1 strains in Argentina, Paraguay, and Uruguay.
HIV subtypes B, F, and BF recombinants have been previously reported in South America. This report describes the presence of HIV-1 subtype C infection in the countries of Argentina, Uruguay, and Paraguay dating back to at least 1999. Surveillance for uncommon non-B/non-F subtype viruses circulating in South America has been conducted in samples obtained from nine countries. Peripheral blood mononuclear cells (PBMC), dried filter paper (FP), and fresh blood (FB) samples were collected from HIV-positive patients from Ecuador, Colombia, Venezuela, Peru, Chile, Bolivia, Argentina, Uruguay, and Paraguay. From a total of 2962 HIV seropositive samples examined during a 9-year period (1995-2003), only 11 (0.4%) were found to be infected with non-B/non-F HIV variants. Eight of these 11 strains were determined to be subtype C by heteroduplex mobility assay (HMA). Five of these 8 strains were further characterized by sequencing and phylogenetic analysis of the protease (Pro) and reverse transcriptase (RT) region of the genome and two were sequenced full length. One of the strains was found to be a unique BC recombinant. The spread of a third subtype of HIV, subtype C, should raise the question of its potential future role in the HIV epidemic in this region.
Adult↗
Regulation of afferent transmission in the feeding circuitry of Aplysia.
Although feeding in Aplysia is mediated by a central pattern generator (CPG), the activity of this CPG is modified by afferent input. To determine how afferent activity produces the widespread changes in motor programs that are necessary if behavior is to be modified, we have studied two classes of feeding sensory neurons. We have shown that afferent-induced changes in activity are widespread because sensory neurons make a number of synaptic connections. For example, sensory neurons make monosynaptic excitatory connections with feeding motor neurons. Sensori-motor transmission is, however, regulated so that changes in the periphery do not disrupt ongoing activity. This results from the fact that sensory neurons are also electrically coupled to feeding interneurons. During motor programs sensory neurons are, therefore, rhythmically depolarized via central input. These changes in membrane potential profoundly affect sensori-motor transmission. For example, changes in membrane potential alter spike propagation in sensory neurons so that spikes are only actively transmitted to particular output regions when it is behaviorally appropriate. To summarize, afferent activity alters motor output because sensory neurons make direct contact with motor neurons. Sensori-motor transmission is, however, centrally regulated so that changes in the periphery alter motor programs in a phase-dependent manner.
Animals↗
[Freezing of gait or freezing of quadruped gait].
Parkinson's disease (PD) is frequently associated with freezing of gait. Patients with advanced PD learn to use tricks to relieve freezing of gait. Recently one of our patients informed us that he could overcome freezing of gait by crawling. Since crawling is a sort of quadruped gait that humans develop transiently, we wondered whether quadruped gait is preserved in PD. We recruited 16 patients with PD who had frequent and disabling freezing of gait. Under experimental conditions, after 12 hours without medication, seven patients developed biped freezing of gait. Of these seven patients, only two patients (the patients with the most severe freezing) also developed quadruped freezing of gait. This experiment suggests that bipedal gait is affected early in PD, while quadruped gait is preserved until late in this disease.
Aged↗
[Accuracy and reliability of the immediate determination of haemoglobin using the HemoCue B hemoglobin in patients undergoing haemodialysis].
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Anemia, Iron-Deficiency↗