Can 3D power Doppler identify levator ani vascularization at its pubic insertion?

INTRODUCTION AND HYPOTHESIS: The levator ani is the most important muscle in the pelvic floor. This pilot study using 3D power Doppler aimed to identify normal vascularization of the puborectalis/pubovisceralis muscle at its insertion in nulliparous and premenopausal women. METHODS: Forty nulliparous and premenopausal women were evaluated at the Gynecological Unit of a tertiary hospital. All women underwent a translabial ultrasound (US) with 3D power Doppler. Pubic insertion of the puborectalis/pubovisceralis muscle was assessed in the axial plane, and vascularization index (VI), flow index (FI), and vascularization-flow index (VFI) were calculated bilaterally. Volumes were analyzed offline by two independent observers, and interobserver agreement and correlation between variables was calculated. RESULTS: A high, statistically significant, correlation was found between the three US indices on each side (p < 0.005). No correlation was found between the US index and either of the demographic parameters of body mass index [(BMI), p > 0.241], or age (p > 0.398). The degree of correlation between the same index on the different sides was not statistically significant. CONCLUSION: Puborectalis/pubovisceralis muscle vascularization at its insertion can be identified using 3D Doppler US, but values differ substantially from those of the contralateral side.

Presence of virulence markers in environmental Vibrio vulnificus strains.

AIMS: This work aims to demonstrate the presence of several genes and factors associated with virulence in strains isolated from the environment at Pueblo Viejo Lagoon, State of Veracruz, Mexico. METHODS AND RESULTS: In this study, we investigated the production of V. vulnificus virulence factors, as cytolysin (haemolysin), RTX toxin, metalloprotease, siderophores, capsular polysaccharide, adhesion structures (like type IV pili), and polar and lateral flagella, involved in swimming and swarming (or, at least, the presence of genes encoding some of them) in 40 strains of V. vulnificus isolated from water and food. The results indicate that strains of environmental origin possess potential virulence characteristics. CONCLUSIONS: Caution should be exercised when consuming raw shellfish (especially by those more susceptible risk groups). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first work focused on the evaluation of V. vulnificus virulence factors in Mexico.

Diversity of culturable halophilic archaea isolated from Rambla Salada, Murcia (Spain).

We have studied the diversity of culturable halophilic Archaea at Rambla Salada, Murcia (south-eastern Spain). We made 8 samplings at different places in this habitat during the years 2006 and 2007 and isolated a total of 49 strains, which were identified by means of phenotypic tests and the hypervariable V1-V3 region of the 16S rRNA gene sequences (around 500 bp). The ribosomal data showed that the isolates belonged to 12 genera within the Halobacteriaceae family, with Haloferax and Natrinema being the most abundant. Five strains showed less than 97% sequence identity with validly described species and may well represent new taxa. All the strains grew best with around 25% w/v salts, required high concentrations of NaCl and magnesium and produced red to pink colonies. They were facultative anaerobes with both respiratory and fermentative metabolisms. The diversity of the archaeal community was analysed with the MOTHUR package. We identified 14 OTUs at the 3% genetic distance level and found quite high diversity. Rarefaction curves of richness estimators and diversity indices demonstrated that our collection of isolates represented the archaeal community at Rambla Salada that can be isolated under the conditions used in this work. This is the first report to be published on the culturable archaea at Rambla Salada, an area of considerable ecological interest.

Contingent screening for Down syndrome completed in the first trimester: a multicenter study.

OBJECTIVE: To assess a new contingent screening strategy for Down syndrome completed in the first trimester. METHODS: First-trimester screening combining nuchal translucency thickness measurement and assessment of serum analytes (combined test) was offered to pregnant women who presented for prenatal care during the first trimester to nine health centers and community hospitals in the area served by the Catalan Public Health Service. If an intermediate risk (1/101-1/1000) for Down syndrome was identified, women were referred to the Hospital Clinic Barcelona for risk reassessment that included the use of secondary ultrasound markers (nasal bone, ductus venosus blood flow and tricuspid flow). Intermediate-risk women were divided into two subgroups for further analysis: high-intermediate risk (1/101-1/250) and low-intermediate risk (1/251-1/1000). We compared feasibility and efficacy of both combined and contingent screening strategies. RESULTS: The combined test, the first screening stage, was performed in 16 001 pregnant women, of whom 1617 (10.1%) were found to have an intermediate risk. Further division of this group showed that 1.8% (n = 289) of women were at high-intermediate risk and 8.3% (n = 1328) at low-intermediate risk. The contingent screening strategy significantly reduced the false-positive rate, from 3.0% to 1.3-1.8% (P < 0.001), without affecting the detection rate (which was 75-79% and 76%, with and without the contingent screening strategy, respectively). However, only 45% of intermediate-risk patients underwent the second screening step due to a preference among high-intermediate-risk (1/101-1/250) women for invasive testing and to low uptake among low-intermediate-risk (1/251-1/1000) women. CONCLUSIONS: The proposed first-trimester contingent strategy reduces the screen false-positive rate without impacting on the detection rate of Down syndrome. The low compliance observed in our study may prevent its use in certain populations.

In vivo transfer of plasmid pRAS1 between Aeromonas salmonicida and Aeromonas hydrophila in artificially infected Cyprinus carpio L.

This study investigated the possible in vivo transfer of plasmid pRAS1 between Aeromonas salmonicida and A. hydrophila inhabiting two different organs of Cyprinus carpio L. To distinguish transconjugants from naturally occurring antibiotic resistant bacteria, twelve luminescent transposon-tagged A. hydrophila strains using mini Tn5luxCDABEKm2 transposon were generated. In conjugal transfer experiments, fish were conditioned with the donor bacteria and subsequently immersed in water containing the recipient strain. Bacteria were recovered from gills and intestines and isolated by growth on selective plates. Transconjugants were identified by their resistance to the pRAS1 encoded antimicrobials and by light emission. In vivo transfer frequencies ranged between 10(-3) and 10(-6) and were somewhat lower in intestines, compared to gills. Transfer frequencies were also smaller relative to those obtained in vitro. The minimal amount of donor and recipient bacteria needed to yield detectable transconjugants in vivo was 1 x 10(4) CFU mL(-1). Implications of this plasmid transfer in natural settings and its possible consequences to human health are discussed.

Characterization of exopolysaccharides produced by three moderately halophilic bacteria belonging to the family Alteromonadaceae.

AIMS: To study the exopolysaccharides (EPSs) produced by three novel moderately halophilic species belonging to the family Alteromonadaceae to optimize EPS yields, characterize their physical and chemical properties and evaluate possible biotechnological applications for these polymers. METHODS AND RESULTS: EPSs synthesized by Idiomarina fontislapidosi F32(T), Idiomarina ramblicola R22(T) and Alteromonas hispanica F23(T) were collected and analysed under optimum conditions: MY medium supplemented with 7.5% (w/v) salts; 32 degrees C; and 1% (w/v) glucose. Polymers were synthesized mainly during the early stationary growth phase with yields ranging from 1 to 1.5 g l(-1). The Idiomarina species each produced an anionic EPS composed mainly of glucose, mannose and galactose. A. hispanica synthesized an anionic EPS composed mainly of glucose, mannose and xylose. Solutions of all the polymers were low in viscosity and pseudoplastic in their behaviour. They showed emulsifying activity and the capacity to bind some metals. CONCLUSIONS: The Alteromonadaceae species studied in this work produced EPSs with physical and chemical properties different from those produced by other halophilic and nonhalophilic bacteria, suggesting that the wide diversity of micro-organisms being encountered nowadays in hypersaline environments offers enormous potential resources for biotechnological applications. SIGNIFICANCE AND IMPACT OF THE STUDY: We have optimized the EPS production and analysed new biopolymers produced by some recently described, moderately halophilic bacteria. These biopolymers are chemically and physically different from others already in use in biotechnology and offer hopes for new applications, especially in the case of A. hispanica, which may prove to be a viable source of xylo-oligosaccharides.

Chitotriosidase activity in goat blood and colostrum.

Chitotriosidase (ChT) activity has not been investigated in ruminants, and therefore, we studied this activity in blood and colostrum of 25 pregnant goats and 60 goat kids. Blood samples were taken from pregnant goats at 3, 2, and 1 d prepartum; at partum; and at 1, 2, 3, and 4 d postpartum. Colostrum samples were obtained by machine-milking at partum and 1, 2, 3, and 4 d postpartum. Goat kid blood was collected at birth and every 7 d thereafter until goats kids were 56 d old. The ChT activity ranged from 2,368 to 3,350 nmol/ mL per hour in goat blood serum, and no statistical differences were detected through time. However, activity tended to decrease from 3 d prepartum to 2 d post-partum. Colostrum ChT activity was 3,912 nmol/mL per hour and 465 nmol/mL per hour on the day of delivery and 4 d postpartum, respectively. Colostrum ChT activity was significantly higher at partum than at any other time. The ChT activity in colostrum was significantly greater at 1 d postpartum than at 2, 3, and 4 d postpartum. Chitotriosidase activity did not differ in colostrum collected on d 2, 3, and 4 postpartum. Chitotriosidase activity in goat kid blood serum ranged from 2,664 to 9,231 nmol/mL per hour at birth and 49 d of life, respectively. Chitotriosidase activity in the blood serum increased with age: at birth, activity was significantly less than at 28, 35, 42, 49, and 56 d postpartum. The maximum ChT activity in blood serum was observed at 49 d postpartum. Activity in 49-d-old kids was significantly greater than that observed in kids at 0, 7, and 14 d postpartum.

Characteristics of bioemulsifier V2-7 synthesized in culture media added of hydrocarbons: chemical composition, emulsifying activity and rheological properties.

The bioemulsifier V2-7 is an exopolysaccharide (EPS) synthesized by strain F2-7 of Halomonas eurihalina and it has the property of emulsifying a wide range of hydrocarbons i.e. n-tetradecane, n-hexadecane, n-octane, xylene mineral light and heavy oils, petrol and crude oil. Characteristics of exopolysaccharide V2-7 produced in media supplemented with various hydrocarbons (n-tetradecane, n-hexadecane, n-octane, xylene, mineral light oil, mineral heavy oil, petrol or crude oil) were studied. Yield production varied from 0.54 to 1.45 g L(-1) according to the hydrocarbon added, in the same way chemical composition, viscosity and emulsifying activity of EPS varied with the culture conditions. Respect to chemical composition, percentage of uronic acids found in exopolymers produced in hydrocarbon media was always higher than that described for V2-7 EPS (1.32%) obtained with glucose. This large amount of uronic acid present could be useful in biodetoxification and waste water treatment. On the other hand, the highest amount of biopolymer was synthesized with mineral light oil, while the most active emulsifiers were those obtained from media added with petrol and n-octane. Furthermore, all EPS were capable of emulsifying crude oil more efficiently than the three chemical surfactants tested as control (Tween 20, Tween 80 and Triton X-100). The capacity of strain F2-7 to grow and produce bioemulsifier in presence of oil hydrocarbons together with the high emulsifying activity and low viscosity power of the biopolymers synthesized in hydrocarbons media could be considered highly beneficial for application of both bioemulsifier and producing strain in bioremediation of oil pollutants.

Short communication: Addition of milk replacer to colostrum whey: effect on immunoglobulin G passive transfer in Majorera kids.

Forty-two Majorera kids (21 males and 21 females) were assigned to 3 groups, a colostrum group (C), a colostrum whey group (CW), and a colostrum whey plus milk replacer group (CWMR). All kids were fed twice on the first day and received 4 g of IgG/kg of body weight. No differences were found in serum IgG among the different treatments. Kid serum IgG concentrations on d 2 were 14.57, 17.25, and 13.32 mg/mL in the C, CW, and CWMR group, respectively. Labor time per animal was higher in the C and CW treatments than in the CWMR group (24.2 +/- 2.3, 20.9 +/- 3.4, and 16.1 +/- 1.5 min, respectively). This new management system may decrease labor costs during the colostrum feeding period.

Comparison of chromaffin cells from several animal sources for their use as an in vitro model to study the mechanism of organophosphorous toxicity.

It had been observed that the chromaffin cells of bovine adrenal medulla contain high levels of neuropathy target esterase (NTE), the esterase whose inhibition and aging is associated with induction of the organophosphorous induced delayed neuropathy. In this study, total esterase and NTE activities, and their inhibition kinetics by OPs are characterized in adrenal medulla of several species in order to find the best source for chromaffin cells. Total esterase activity in membrane fraction of bovine, equine, porcine, ovine and caprine were 6100+/-840, 4200+/-270, 5000+/-120, 28800+/-3000, and 10800+/-2400mU/gtissue, respectively (mean+/-S.D., n=3-4). NTE represented around 70%, 24%, 58%, 10% and 24% of the total esterases in the same tissues, respectively. It was deduced that NTE represents between 69% and 89% of the "B-activity" (activity resistant to 40microM paraoxon) in the membrane fraction of all species. The mipafox I(50) calculated for 30-min inhibition of NTE at 37 degrees Celsius ranged between 7.4 and 12microM. These values are in the range of that for brain NTE in hen (the usual model for testing OP delayed neurotoxicity). Considering that bovine adrenal medulla contains high NTE activity, that it represents a high proportion of total activity, it is easier to dissect than adrenal medulla from equine, caprine or ovine, and is more readily available than species cited previously, and that its inhibitory properties are similar to the classical hen brain model, it is deduced that bovine adrenal medulla is the most appropriate source of chromaffin cells to study OP toxicity, with porcine as the second alternative. The kinetic properties of chromaffin cell cultures from bovine and porcine were in accordance with their properties in homogenate and subcellular fractions, and they displayed an appropriate stability and viability of the primary culture to be used in in vitro toxicological studies for both mechanistic and testing purposes.

Transposon mutagenesis in Halomonas eurihalina.

We have established a transposon mutagenesis procedure for the moderate halophile Halomonas eurihalina, a bacteria that produces an exopolysaccharide (EPS) of considerable biotechnological interest. We used suicide plasmids pUT and pSUP102 to introduce the transposons mini-Tn5 and Tn1732 into H. eurihalina via Escherichia coli mediated conjugation. Southern hybridization analysis demonstrated that insertions of the transposon mini-Tn5 into H. eurihalina occurred randomly at single sites in the chromosome, whereas Tn1732 insertion also took place at random, but simultaneously, at several sites. Phenotypic analysis revealed that different mutants were generated by using mini-Tn5. The isolation of exopolysaccharide-defective strains is the first stage towards carrying out genetic studies on EPS production by this microorganism.

Influence of salt concentration on the cellular fatty acid composition of the moderately halophilic bacterium Halomonas salina.

The cellular fatty acid composition of Halomonas salina, a moderately halophilic bacterium grown at different salt concentrations, is reported. Fatty acids C16:0 and C18:1 were major components and significant amounts of C16:1, C18:0 and cyc-C19:0 were also detected. The results showed clear chemotaxonomic relationships with recognized members of the genus Halomonas. The salt concentration greatly influenced the fatty acid composition, suggesting activation of cyclopropane synthetase at high levels of salt, since increases in cyclopropane fatty acids with decreases in monounsaturated fatty acids were observed as the salt concentration in the medium rose.

Effect of external salinity changes on cellular composition of some ions and amino acids in Deleya halophila.

Changes in total cellular composition of some ions and amino acids of the moderate halophile Deleya halophila were studied in response to external salinity changes. Among the 14 amino acids investigated, the cellular glycine and aspartic acid content increased with increasing salinity. D. halophila also accumulated Na+ at the highest external salt concentrations.

Bovine chromaffin cell cultures as model to study organophosporus neurotoxicity.

Based on the high level of phenyl valerate esterase activities, and in particular of neuropathy target esterase (NTE) found in bovine adrenal medulla, chromaffin cells culture have been proposed as an alternative model for the study of organophosphorus neurotoxicity. Organophosphorus-induced polyneuropathy is a syndrome related to the inhibition and further modification by organophosphorus compounds of NTE (a protein that displays phenyl valerate esterase activity resistant to mipafox and sensitive to paraoxon). Total phenyl valerate esterase activities found in homogenate, particulate and soluble fractions of bovine adrenal medulla were 5200+/-35, 5000+/-280 and 1700+/-260 mU/g tissue, respectively. Cultured chromaffin cells displayed a total hydrolysing activity of 41+/-5 mU/10(6) cells. Homogenates of bovine adrenal medulla displayed only about 6% of activity sensitive to paraoxon. Most of the phenyl valerate esterase activity inhibited by mipafox (a neuropathy inducing compound) was found in particulate fraction. Cultured chromaffin cells displayed kinetics of inhibition by mipafox similar to the kinetics displayed by homogenates of bovine adrenal medulla. We conclude that NTE could be assayed in this system by only using one inhibitor (mipafox) instead of two (paraoxon and mipafox). Also, the proposal is supported of using chromaffin cells as in vitro model for the study of the role of NTE and related esterases in organophosphorus-induced polyneuropathy.

[Current knowledge on the strain typing of the pathogenic fungus Histoplasma capsulatum var. capsulatum: a review of the findings]. / Estado actual de la tipificación del hongo patógeno Histoplasma capsulatum var. capsulatum: una revisión de los hallazgos.

The classification of microbial strains is currently based on different typing methods, which must meet certain criteria in order to be widely used. Phenotypic and genotypic methods are being employed in the epidemiology of several fungal diseases. However, some problems associated to the phenotypic methods have fostered genotyping procedures, from DNA polymorphic diversity to gene sequencing studies, all aiming to differentiate and to relate fungal isolates or strains. Through these studies, it is possible to identify outbreaks, to detect nosocomial infection transmission, and to determine the source of infection, as well as to recognize virulent isolates. This paper is aimed at analyzing the methods recently used to type Histoplasma capsulatum, causative agent of the systemic mycosis known as histoplasmosis, in order to recommend those that yield reproducible and accurate results.

[Simple-blind clinical study with sublingual lorazepam vs. injectable diazepam as premedication in colonoscopies]. / Estudio clínico simple ciego con lorazepan sublingual vs. diazepan inyectable como premedicación en colonoscopías.

Those patients premedicated at least 90 minutes before coloscopy, with sublingual lorazepam, showed improved sedative effect, better tolerance and collaboration? Less discomfort and pain when air was, as usually insuflated, as compared with patients receiving intramuscular diazepam. Doctor's opinion coincided with the before mentioned preference for sublingual lorazepam prescribed at least 90 minutes before the endoscopic procedure. No adverse side effects were pointed out.

[Endoscopic cholangiography in biliary tract diseases in HIV+ patients]. / La colangiografía endoscópica en afecciones de la vía biliar en pacientes HIV+.

UNLABELLED: Biliary tract involvement in the course of the infection by HIV was first described in 1983. Since then, various opportunistic infections have been responsible of different biliary lesions, coming to a new entity called "cholangiopathy associated to HIV". Our aim was to determined the use of ERCP in the management of HIV patients with cholestasis. From August 1994 to October 1997, 11 HIV patients (8 men, 3 women, mean age 35 y.) were submitted to ERCP because of jaundice (n = 8), upper right abdominal pain (n = 8), fever (n = 6), pruritus (n = 5) and elevated alkaline phosphatase (n-10). In 7 the diagnosis of AIDS had already be made. All had hepatobiliary ultrasound and endoscopic periampullar duodenal mucosa biopsy was taken in 7. According to Cello, 4 types of radiologic lesions were considered: 1) Papillary stenosis with dilated extrahepatic biliary tract. 2) Sclerosing cholangitis (focal intra or extrahepatic stenosis and dilatations). 3) Association of types 1 and 2.4) Choledocal long stenosis in the absence of previous biliary surgery or chronic pancreatitis. Five patients (45%) had biliary abnormalities; in 3 related to HIV infection: sclerosing cholangitis (n = 2) and papillary stenosis (n = 1). Two had choledocal stones. Four had upper right abdominal pain and dilated bile ducts at ultrasound. Cryptosporidium was found in duodenal mucosa in one patient with sclerosing cholangitis and in the patient with papillary stenosis. Biliary stents were placed without sphincterotomy in 2, with relief of pain and improving of cholestasis in only one. The choledocal stones were removed endoscopically in one patient and by surgery in the other. CONCLUSION: The ERCP is a useful method in the diagnosis and treatment of the biliary tract abnormalities associated to HIV.

[Morphologic and physiologic characteristics of Costa Rica pathogenic fungi (Dermatiaceae)]. / Características morfológicas y fisiológicas de hongos patógenos en Costa Rica (Dematiaceae).

Forty one strains of dematiaceous fungi from the Mycology collection of the University of Costa Rica were studied. Thirty three were pathogenic (Fonsecaea pedrosoi, Cladosporium carrionii, Xylohypha bantiana, Exophiala jeanselmei, Rhinocladiella aquaspersa, Phialophora verrucosa) and the other eight were contaminants (Hormodendrum sp.). Morphological studies were done using the slide culture technique. The physiological criteria used were: urease production, gelatin and Loeffler media liquefaction; xanthine, tyrosine, starch and casein hydrolysis; nitrate utilization; carbohydrate uptake; sensitivity to cycloheximide and thermotolerance in glucose-Sabouraud medium. The physiological tests did not provide characteristic patterns for the different genera of pathogenic fungi, even though these differences were detected in non pathogenic fungi; the tests may be useful for the quick separation of both groups. Physiological test may have a limited value in the identification of fungi and the morphological analysis cannot be substituted by physiological studies.

Entry inhibitors directed towards glycoprotein gp120: an overview on a promising target for HIV-1 therapy.

In spite of the unquestionable positive impact of HAART in the treatment of HIV infection, the discovery and development of novel agents directed towards other targets of the replicative cycle of the virus that differ from those targeted by the clinically approved drugs, emerges nowadays as an imperative need. The blockade of HIV entry is a highly promising strategy against the pathogen and glycoprotein gp120 is a central actor in this process. This review discusses the current status in the research of anti-HIV agents targeting specifically the envelope protein gp120. The diverse approaches devoted to the achievement of therapeutic agents against gp120 currently under study are organized and analyzed critically according to their specific mechanism of inhibition and structural features.