[Outbreak of multiresistant and extended spectrum ß-lactamase producing Klebsiella pneumoniae in a high risk neonatal unit]. / Brote por Klebsiella pneumoniae multiresistente y productora de ß-lactamasa de espectro extendido en una unidad de alto riesgo neonatal.

Klebsiella pneumoniae as a producer of extended spectrum beta-lactamase (ESBL) has played an important role as a cause of infection in the neonatal high risk unit (NHRU) of the Autonomous Hospital Institute of the Universidad de Los Andes (AHIULA). In this paper an outbreak caused by this bacterial specie that affected neonates hospitalized in this unit during February 2007 is described. Besides, the environment and the personnel were studied as possible sources of this organism. The strains of K. pneumonia isolated from the outbreak had the same resistance phenotype, produced ESBL type TEM and SHV and belonged to the same genotype as the isolated strains from the hands and the soapy solutions, possible sources of infection. This indicates that it was the same clone. The outbreak was resolved using two important measurements: reinforcing hand washing and with the opportune treatment of neonates with imipenem.

Brote por Klebsiella pneumoniae multiresistente y productora de β-lactamasa de espectro extendido en una unidad de alto riesgo neonatal / Outbreak of multiresistant and extended spectrum β-lactamase producing Klebsiella pneumoniae in a high risk neonatal unit

Klebsiella pneumoniae productora de β-lactamasa de espectro expandido (BLEE) ha jugado un papel importante como causa de infecciones en la unidad de alto riesgo neonatal (UARN) del Instituto Autónomo Hospital Universitario de Los Andes (IAHULA). En el presente trabajo se describe un brote ocasionado por esta bacteria en los neonatos hospitalizados en dicha unidad durante el mes de febrero 2007, así como también, cepas aisladas en los meses siguientes al brote y además, se estudia el ambiente y el personal, como posible fuente de esta bacteria. Las cepas de K. pneumoniae aisladas del brote eran del mismo fenotipo de resistencia, productoras de (3LEE tipo TEM y SHV y pertenecían al mismo genotipo que las cepas aisladas de las manos y de las soluciones jabonosas, posible fuente de infección, lo cual indica que se trataba del mismo clon. El brote se resolvió usando dos importantes medidas: reforzando el lavado de manos y con la indicación oportuna de imipenem a los neonatos afectados.

Klebsiella pneumoniae as a producer of extended spectrum beta-lactamase (ESBL) has played an important role as a cause of infection in the neonatal high risk unit (NHRU) of the Autonomous Hospital Institute of the Universidad de Los Andes (AHIULA). In this paper an outbreak caused by this bacterial specie that affected neonates hospitalized in this unit during February 2007 is described. Besides, the environment and the personnel were studied as possible sources of this organism. The strains of K. pneumonia isolated from the outbreak had the same resistance phenotype, produced ESBL type TEM and SHV and belonged to the same genotype as the isolated strains from the hands and the soapy solutions, possible sources of infection. This indicates that it was the same clone. The outbreak was resolved using two important measurements: reinforcing hand washing and with the opportune treatment of neonates with imipenem.

Intracerebroventricular insulin-like growth factor-1 decreases feeding in diabetic rats.

Insulin-like growth factor-1 (IGF-1) is a hormone that is important in the regulation of growth processes and additionally has been demonstrated to modulate metabolic and autonomic responses. Some of its effects are mediated by the central nervous system (CNS), and there are IGF-1 receptors dispersed throughout the CNS. Both IGF-1 and insulin alter peripheral metabolic and autonomic nervous activity by a central mechanism, and the well-defined role of insulin in the regulation of feeding, especially in diabetes, led us to investigate the effect of chronic central administration of IGF-1 on metabolic and feeding parameters in normal and diabetic rats. Normal and diabetic rats with intracerebroventricular cannulas were given IGF-1, insulin (0.5 nmol/animal), or artificial cerebrospinal fluid via cannula twice daily for 4 d. Blood samples were collected on d 2 and 4, and the body weights and food intake were recorded daily. IGF-1 administered intracerebroventricularly did not alter plasma glucose, insulin, body weight, or food intake in normal rats. However, in diabetic animals, IGF-1 decreased food intake but did not alter blood glucose or plasma insulin. In correlated studies, intracerebroventricular insulin decreased food intake in both normal and diabetic animals. From these studies, we conclude that IGF-1 may act centrally to decrease food intake in the hyperphagic diabetic animals but not in normal animals. This suggests that diabetic animals have an increased sensitivity to CNS IGF-1.

The effect of diabetes and sex on nitric oxide-mediated cardiovascular dynamics.

Diabetes is associated with impaired cardiovascular responses that are especially prominent in females. Since nitric oxide (NO)-mediated effects on cardiovascular dynamics are altered in diabetes, we evaluated the effect of L-NAME, a nitric oxide synthase (NOS) antagonist, on mean arterial pressure (MAP), heart rate (HR), and selective vascular flows in both male and female normal and diabetic rats as an index of NO activity. Rats were made diabetic using streptozotocin and maintained for 5-6 weeks. Following anesthesia with urethane/alpha-chloralose, the femoral artery and vein were cannulated for recording and sampling, and flow probes were placed on the iliac, renal, and superior mesenteric arteries. A bolus infusion of L-NAME (10mg/ kg) resulted in a rapid +52% and +68% increase in MAP in normal female and male rats, respectively. However, diabetic females' and males' responses were significantly lower (44% and 45%, respectively) when compared with their normal counterparts. The decreased HR in response to the peak pressor effect of L-NAME was more prominent in normal females compared with normal males (-14% vs 2%). The results in diabetic females and males were equivalent (-6% vs -9%, respectively). L-NAME decreased the conductance (flow/MAP) an average of 65% in all three vascular beds in normal female rats. In diabetic females, the iliac and superior mesenteric responses to L-NAME were less, and the renal conductance was contrastingly increased 23%. The response to L-NAME was comparable (-62%) in the renal and superior mesenteric and less (-40%) in the iliacs of normal versus diabetic males. We concluded that diabetes is associated with a decreased pressor response to NOS inhibition. And the impaired constriction response of the renal vessels noted in female diabetic rats may provide a basis for the increased renal pathology observed in diabetic humans.

Autonomic and endothelial dysfunction in experimental diabetes.

The effect of streptozotocin induced diabetes on autonomic regulation of heart rate and endothelial function was examined in Sprague-Dawley rats. Weanling rats (3-4 weeks of age) of either sex were randomly assigned to a non-diabetic (male 5, female 6) or diabetic (male 4, female 5). Diabetes was induced with a single intraperitoneal (IP) injection of streptozotocin (STZ, 100 mg/kg). Nondiabetic rats received an IP injection of saline. Eight weeks after injection, rats were chronically instrumented with a left jugular venous catheter and a left carotid arterial catheter. After recovery (5 days) cardiac sympathetic tonus, parasympathetic tonus and intrinsic heart rate were determined. On an alternative day, the pressor response to nitric oxide synthase inhibition (NOS-X) was determined in areflexic rats. Cardiac sympathetic tonus (72 +/- 13 vs. 41 +/- 7), parasympathetic tonus (-51 +/- 10 vs. -22 +/- 7), and intrinsic heart rate (368 +/- 6 vs. 292 +/- 9), were reduced in diabetic rats. Furthermore, diabetic rats had a smaller pressor response (A33 +/- 7 vs. A66 +/- 5) to NOS-X. These results document impaired autonomic control of heart rate and endothelial dysfunction in 8-week streptozotocin induced diabetic rats.

Molecular basis of extended-spectrum beta-lactamase production in nosocomial isolates of Klebsiella pneumoniae from Mérida, Venezuela.

Twelve Klebsiella pneumoniae isolates resistant to expanded-spectrum cephalosporins and aztreonam, from patients with nosocomial septicaemia at the intensive care unit of the Andes University Hospital, Mérida, Venezuela, were studied for production of extended-spectrum beta-lactamase (ESbetaL) activity. All were also resistant to gentamicin, kanamycin, tetracycline and chloramphenicol but sensitive to cefoxitin, imipenem, amikacin and tobramycin. Production of ESbetaL activity was confirmed by restoring susceptibility to ceftazidime in the presence of clavulanic acid. All isolates carried an identical plasmid of approximately 87 kb. Resistance to beta-lactams, aminoglycosides, tetracycline and chloramphenicol was lost en bloc after plasmid curing by treatment with acridine orange and was transferable en bloc to Escherichia coli by conjugation. Transconjugants always showed the same plasmid profile as that of Klebsiella donors. Isoelectric focusing analysis of the crude extracts of transconjugants showed in all cases, the presence of two beta-lactamases of pI 5.4 and 8.2. Analysis of the plasmid carried by one of the transconjugants by means of hybridization assays, revealed the presence of both bla(TEM) and bla(SHV) determinants. Cloning and sequencing of each determinant identified them as bla(TEM-1) and bla(SHV-5), respectively, the latter being responsible for the ESbetaL activity. Results of this study indicate that ESbetaL determinants of the SHV-type carried by transferable elements, are spreading among nosocomial isolates of K. pneumoniae in Mérida, Venezuela.

Arterial baroreflex regulation of regional vascular conductance at rest and during exercise.

We tested the hypothesis that dynamic exercise resets the operating point and attenuates the spontaneous gain of the arterial baroreflex regulation of mesenteric and hindlimb vascular conductance in hypertensive rats. Eleven adult male spontaneously hypertensive rats were chronically instrumented with left carotid arterial catheters and Doppler ultrasonic flow probes around the superior mesenteric and left common iliac arteries. After the rats recovered, arterial baroreflex function was examined by recording reflex changes in conductance in response to spontaneous changes in mean arterial pressure before exercise and during steady-state treadmill running at 6 and 18 m/min. Dynamic exercise reduced the spontaneous baroreflex gain of mesenteric conductance (by 51 and 36%) and maximum mesenteric conductance (by 24 and 32%) at 6 and 18 m/min, respectively. In sharp contrast, dynamic exercise increased the spontaneous maximum iliac conductance (by 32 and 47%) without changing the spontaneous gain. Sinoaortic denervation eliminated the relationship between mean arterial pressure and conductance by reducing the mesenteric (92%) and iliac (68%) vascular conductance gain. These results demonstrate that dynamic exercise has differential effects on the regulation of mesenteric and iliac vascular conductance in hypertensive rats.

Vascular dilatatory responses to sodium nitroprusside (SNP) and alpha-adrenergic antagonism in female and male normal and diabetic rats.

Diabetes is associated with impaired vascular dilatatory responses that appear to be influenced by sex as well as diabetic state. Therefore, we hypothesized that vascular and sympathetic control function exhibit a greater deterioration following the induction of diabetes in female than in male rats. We conducted a comparative determination of the effect of sodium nitroprusside (SNP, a nitrous oxide donor) and that of an alpha1-adrenergic antagonist, prazosin, on selective vascular flows, mean arterial pressure (MAP), and heart rate (HR), in female and male normal and diabetic rats. Rats were made diabetic using streptozotocin (50 mg/kg, iv) and maintained for 5-6 weeks. Following anesthesia with urethane/alpha-chloralose, the femoral artery and vein were cannulated for recording and sampling. Flow probes were placed on the iliac, renal, and superior mesenteric arteries. SNP (1, 5, 10, and 20 microg/kg) infusions resulted in a dose-dependent decrease in MAP in normal and diabetic rats. The decrease in MAP in normal males was 37% less at the 20 microg/kg concentration of SNP when compared to normal females. The HR was not significantly changed in response to the hypotensive effect of SNP; however, reflex tachycardia was more prominent in diabetic males. The vascular conductance (flow/MAP) was increased by SNP in normal and diabetic rats in a dose-dependent fashion; however, the responsiveness was decreased in the iliac and superior mesenteric and increased in the renal arteries in diabetics when compared to normals. Diabetic males were 42% and 28% less responsive to SNP in the iliac and superior mesenteric arteries, respectively. On the other hand, diabetic females were 1.5-fold more responsive in the renal artery when compared to normals. Prazosin (4 mg/kg) decreased the MAP in normal and diabetic rats to a comparable degree. Prazosin increased the vascular conductance in all three vascular beds in normal and diabetic rats with the greater increase occurring in the iliac (118%) and superior mesenteric (110%) arteries. We concluded that diabetes is associated with an increased response to NO in the renal vessels and a decreased response in the iliac and superior mesenteric vessels in both females and males. alpha-Adrenergic tone was greatest in diabetic female and male rats. This study suggests that decreased vascular flow in diabetes is a result of a combination of decreased sensitivity to NO and increased adrenergic tone.

[Clinical, epidemiological and microbiological aspects of Mobiluncus sp. in bacterial vaginosis]. / Aspectos clínicos, epidemiológicos y microbiológicos de Mobiluncus sp. en la vaginosis bacteriana.

BACKGROUND: In this paper, our goal was to determine the optimal isolation conditions, biochemical characterization, and preservation of species of the genus Mobiluncus, associated with bacterial vaginosis in patients attending the family planning clinic. Also, we tried to relate its presence with demographic variables and criteria used in the clinical diagnosis of bacterial diagnosis. METHODS: The specimen from the posterior fornix were collected and transported to the laboratory in a Stuart medium, one at room temperature and the other at 4 degrees C. These samples were inoculated in anaerobic culture media. RESULTS: Of a total of 92 patients studied, 61 (66.3%) were normal, 28 (30.4%) bacterial vaginosis, and 3 (3.3%) had intermediate vaginosis. There was statistically significant relationship only with intrauterine device use (p = 0.00499). The presence of curved rod, using Gram's method, was significantly related with pH (p = 0.00000) positive amines test (p = 0.00000), and the presence of clue cells (p = 0.00000). Mobiluncus was observed in 23 samples (82%), and the majority (15) using RLK agar (cold enrichment technique). With conventional techniques, we identified 12 strains as Mobiluncus curtisii and 3 strains as Mobiluncus mulieris. The strains of Mobiluncus sp. grew better from litmus milk conserved at -30 degrees C. CONCLUSION: Isolating Mobiluncus sp. is fairly easy, if the right media and the techniques are used.

[Characterization of plasmids which mediate resistance to multiple antibiotics in gram-negative bacteria of nosocomial origin]. / Caracterización de plásmidos que median la resistencia a múltiples antibióticos en bacterias gramnegativas de origen nosocomial.

BACKGROUND: The genetic and molecular mechanisms involved in antimicrobial resistance of 10 strains of gramnegative bacilli (1 Serratia marcescens; 2 Escherichia coli; 1 Proteus mirabilis; 4 Klebsiella pneumoniae; 1 Enterobacter cloacae y 1 Alcaligenes faecalis), isolated from adult patients with nosocomial pulmonary infection at the in-patient facilities of the University Hospital of Los Andes, Mérida, Venezuela, have been studied. METHODS: The antimicrobial susceptibility was determined by minimum inhibitory concentrations using the dilution method in agar. The study of extrachromosomal genes was carried out by conjugation, bacterial infection with the bacteriophage M13 and curing of plasmid by acridine orange. The plasmids were isolated by alkaline lysis and analysis of restriction endonuclease digestion was carried out separately using the enzymes EcoRI and HindIII. A DNA probe, derived from the region which encodes the TEM-1 beta-lactamase of the plasmid pBR322 was used for dot-blot hybridization tests. RESULTS: All of the gramnegative bacilli showed resistance to ampicillin, carbenicillin and cephalothin (> 128 micrograms/ml) and 3 strains also showed resistance to gentamicin (> 64 micrograms/ml). Genetic and molecular procedures showed the presence of conjugative plasmids of approximately 54 kb in all the 10 strains. The restriction patterns obtained by using EcoRI and HindIII indicated common DNA fragments in most of the plasmids studied. The dot-blot hybridization tests confirmed homology between the plasmids and the DNA probe used (TEM-1 beta-lactamase). CONCLUSIONS: In this study, the gramnegative bacteria of nosocomial origin harbored self-transferable plasmids of approximately 54 kb, which mediate resistance to gentamicin and encode a beta-lactamase of the TEM group.

[Comparative study of 4 methods for identifying species of the genus Aeromonas]. / Estudio comparativo de 4 métodos para la identificación de especies del género aeromonas.

It is given an explanation of the results obtained with the following methods: AEROKEY II, AEROKEY II + Abbot's scheme, Api 20 NE, and the Biolog System. The study was conducted with 38 strains of Aeromonas isolated from children under 5 with acute diarrheal disease (ADD). The AEROKEY II + Abbot's scheme proved to be the best identification method.

Especies de aeromonas asociadas a diarrea: características microbiológicas y clínicas / Species of aeromonas associated a diarrhea: characteristic microbiological and clinical

Para estudiar las características microbiológicas y clínicas de Aeromonas sp. asociada a Enfermedad Diarreica Aguda (EDA) se analizaron, desde junio de 1993 1 diciembre de 1994, 397 muestras de heces de niños menores de cinco años con EDA y 121 asintomáticos. Fueron aisladas en una frecuencia de 11,8 y 5,8 por ciento en los casos de EDA y control, respectivamente. A.hydrophila y A.caviae fueron las especies más aisladas en ambos casos. Como patógenos únicos se encontraron en un 38,3 por ciento, y asociados en un 61,7 por ciento. La presentación del cuadro clínico fue 50 por ciento de tipo disentérico: 33,33 por ciento de tipo secretorio y 16.66 por ciento de tipo indeterminado. El mejor medio de cultivo fue el DNA-ampicilina. Se obtuvo un 1 por ciento con producción de H2S y un 5 por ciento fermentadoras de la lactosa. En las cepas se detectó por lo menos dos de las enzimas estudiadas; en base a esto, existe una importante circulación de este género con capacidad de producir EDA

Nitric oxide regulation of superoxide-dependent lung injury: oxidant-protective actions of endogenously produced and exogenously administered nitric oxide.

The influence of endogenous cell .NO production and .NO derived from exogenous sources on oxidant injury to cultured fetal rat lung alveolar epithelium and an animal model of pulmonary oxidant injury was examined. Confluent fetal rat alveolar epithelial cell monolayers were stimulated to produce .NO after treatment with a combination of cytokines (IL-1 beta, TNF-alpha, IFN-gamma), LPS and zymosan-activated serum (CZ). Cell injury, assessed by 14C-adenine release, was significantly increased compared to basal and CZ-induced cells after inhibition of .NO synthesis by L-NMMA. Cell monolayer macromolecule barrier function was determined by the rate of diffusion of 125I-albumin from the apical to basolateral side of monolayers. Following exposure to CZ and/or O2.- generated by xanthine oxidase + lumazine (XO), endogenous cell .NO production and exogenously administered .NO (from .NO donors S-nitrosyl-glutathione and S-nitroso-N-acetylpenicillamine) significantly inhibited the increased monolayer permeability induced by exposure to reactive oxygen species. Furthermore, inhalation of 5-10 ppm of .NO significantly reduced the toxicity of > 95% oxygen to adult rats. We conclude that when cultured pulmonary epithelial cells and lung tissue in vivo are subjected to inflammatory mediators or acute oxidative stress, .NO can play a protective role by inhibiting O2.(-)-dependent toxicity.

Interaction of surfactant mixtures with reactive oxygen and nitrogen species.

Increased concentrations of partially reduced oxygen and nitrogen reactive species damage the alveolar epithelium and either cause or exacerbate surfactant deficiency. For this reason, there is a quest to identify surfactant replacement mixtures, which in addition to repleting depleted surfactant stores can also reduce the steady-state concentrations of reactive species in the alveolar space. Herein, we evaluated the ability of natural lung surfactant (NLS) and two mixtures (Exosurf and Survanta) used clinically for the correction of surfactant deficiency to scavenge hydroxyl radical-type species (.OH), generated either by the decomposition of peroxynitrite or by Fenton reagents (FeCl3 + H2O2). Exosurf or Survanta decreased .OH only when present at high lipid concentrations (6.5 mM). On the other hand, 40 microM of NLS decreased .OH concentrations from 75 +/- 2 to 53 +/- 2 microM (P < 0.05), most likely because of the interaction of .OH with protein sulfhydryl groups. Similarly, 40 microM of NLS incubated with a bolus of H2O2 (400 microM) decreased the H2O2 concentration in the supernatant by approximately 50%, due to the presence of catalase-type activity. In contrast to NLS, neither Exosurf nor Survanta scavenged H2O2, even when present at millimolar lipid concentrations. We concluded that Exosurf and Survanta contain limited antioxidant activity compared with NLS.

Bacterias anaeróbicas como agentes etiológicos de infecciones en el ganado vacuno / Anaerobic bacteria as etiology agent in infections in the cattle

Se estudiaron 14 muestras clínicas provenientes de ganado vacuno de las Haciendas El Joque y La Atalaya, Mérida, con el propósito de aislar bacterias anaeróbicas involucradas en procesos infecciosos que afectan al ganado y que muchas veces son causas de pérdida económicas en las explotaciones lecheras de la región. En 11 casos, de los 14 estudiados, se aislaron bacterias anaeróbicas. Fusobacterium necrophorum fue el micro-organismo anaerobio más frecuente aislado, asociado con otras bacterias anaeróbicas y facultativas anaerobias como Corynebacterium (Actinomyces) pyogenes. Bacteroides pigmentados fue aislado de los casos de pododermatitis estudiados, Clostridium perfrigens de un cuadro clínico de enterotexemia fulminante en un becerro y Actinomyces sp de muestras clínicas de un absceso en la región cervical de uno de los ejemplares estudiados. Los resultados obtenidos nos permiten presumir que probablemente la incidencia de estas infecciones por bacterias anaeróbicas sea alta y que muchas veces pueden pasar desapercibidas si no se hace un diagnóstico micro-biológico. Por lo tanto, es de gran interés para el veterinario tener conocimiento de estas infecciones para prevenirlas y controlarlas

Properties of a bacteriocin produced by Clostridium bifermentans 18137.

Una particula filamentosa, con um amplio espectro de actividad similar a una bacteriocina, producida por Clostridium bifermentans 18137 fue caracterizada parcialmente. La bacteriocina inhibio el crecimiento de Clostridium aerofoetidum y causo lisis de celulas viables, pero no de celulas previamente muertas por exposicion al aire. La particula fue precipitada por sulfato de amonio a 40% de saturacion, tuvo una densidad entre l,22 y 1,29 g/ml y consistio en filamentos helicoidales. Fue inactivada por pronasa, quimotripsina y calentamiento a 95o.C por 5 min. R Nasa, Ba++ y Cd++ inhibieron las lisis cuando se anadieron junto con la bacteriocina, pero se mantuvo la actividad inhibitoria bajo las mismas condiciones. Se encontro RNA asociado con la bacteriocina y fue removido solo parcialmente por tratamiento con R Nasa. Celulas de C. aerofoetidum tratadas con la bacteriocina sufreiron dano extenso que incluyo destruccion de la membrana citoplasmatica y la pared celular, y disolucion del contenido celular. Tambien se observo la formacion de esferoplastos y fantasmas bacterianos

Growth inhibition activity and bacteriophage and bacteriocinlike particles associated with different species of Clostridium.

Thirty-four strains corresponding to 16 species of Clostridium were examined for growth inhibition activity and production of phage and bacteriocinlike particles. Twenty-five strains were found to inhibit growth of one or more of the other strains. The widest range of activity corresponded to C. bifermentans 18137 which inhibited growth of 23 strains. Cell-free filtrates of positive cultures were inactive with the exception of the one from C. bifermentans 18137. A wide variety of particles with diverse morphology was observed in lysates of cultures treated with mitomycin C (1 microgram/mL).