Field enhancement in a doubly resonant optical parametric oscillator.

Single-resonant and (signal/idler) double-resonant optical parametric oscillators differ fundamentally on the conversion efficiency back to the pump wave. The nonpresent idler in the single-resonant case allows for signal intracavity enhancement well beyond the pump power level. This paper answers the question, how the phase-matched back conversion in a doubly-resonant system can be overcome to reveal substantial power enhancement, and what parameters it depends on. In a degenerate double-resonant OPO (DROPO) pumped by a thin-disk oscillator, an enhancement up to a factor of four is shown experimentally. Support of a semianalytical theory is presented with exceptionally simple relations between enhancement and intracavity losses. Interestingly, our theory predicts no fundamental limit to the maximal field enhancement or conversion efficiency.

Serotypes, virulence markers and cell invasion ability of Shiga toxin-producing Escherichia coli strains isolated from healthy dairy cattle.

AIM: The occurrence of virulence markers, serotypes and invasive ability were investigated in Shiga toxin-producing Escherichia coli (STEC) isolated from faecal samples of healthy dairy cattle at Rio de Janeiro State, Brazil. METHODS AND RESULTS: From 1562 stx-positive faecal samples, 105 STEC strains were isolated by immuno-magnetic separation (IMS) or plating onto MacConkey agar (MC) followed by colony hybridisation. Fifty (47·6%) strains belonged to nine serotypes (O8:H19, O22:H8, O22:H16, O74:H42, O113:H21, O141:H21, O157:H7, O171:H2 and ONT:H21). The prevalent serotypes were O157:H7 (12·4%), O113:H21 (6·7%) and O8:H19 (5·7%). Virulence genes were identified by polymerase chain reaction (PCR). E-hlyA (77·1%) was the more prevalent virulence marker, followed by espP (64·8%), saa (39%), eae (24·8%) and astA (21·9%). All O157:H7 strains carried the γ (gamma) variant of the locus of enterocyte effacement (LEE) genes and the stx2c gene, while the stx1/stx2 genotype prevailed among the eae-negative strains. None of the eae-positive STEC produced the localized adherence (LA) phenotype in HEp-2 or Caco-2 cells. However, intimate attachment (judged by the fluorescent actin staining test) was detected in some eae-positive strains, both in HEp-2 (23·1%) and in Caco-2 cells (11·5%). Most strains (87·5%) showed 'peripheral association' (PA) adherence phenotype to undifferentiated Caco-2 cells. Twenty-five (92·6%) of 27 strains invaded Caco-2 cells. The highest average value of invasion (9·6%) was observed among the eae-negative bovine strains from serotypes described in human disease. CONCLUSION: Healthy dairy cattle is a reservoir of STEC carrying virulence genes and properties associated with human disease. SIGNIFICANCE AND IMPACT OF THE STUDY: Although reports of human disease associated with STEC are scarce in Brazil, the colonization of the animal reservoir by potentially pathogenic strains offers a significant risk to our population.

The influence of microhabitat on the population dynamics of four herbaceous species in a semiarid area of northeastern Brazil.

Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.

The influence of microhabitat on the population dynamics of four herbaceous species in a semiarid area of northeastern Brazil / A influência dos microhabitats sobre a dinâmica de populações de quatro espécies herbáceas em uma área semiárida no nordeste do Brasil

Abstract Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.

Resumo A variação anual na precipitação é considerada o fator mais importante que influencia a dinâmica populacional em ambientes secos. No entanto, diferentes fatores podem controlar a dinâmica populacional em diferentes microhabitats. A hipótese deste estudo é que as variações de microhabitats podem atenuar a influência da sazonalidade climática sobre a dinâmica populacional de espécies herbáceas em uma áreas floresta seca (Caatinga) no Brasil. Então, objetivou-se avaliar a influência de três microhabitats (plano, rochoso e ciliar) sobre a dinâmica das populações de quatro espécies herbáceas (Delilia biflora, Commelina obliqua, Phaseolus peduncularis e Euphorbia heterophylla) em um fragmento de Caatinga da Estação Experimental do Instituto de Pesquisas Agronômicas de Pernambuco, no Brasil, durante um período de três anos. D. biflora, C. obliqua e P. peduncularis foram encontrados em todos os microhabitats, mas elas estavam presentes em baixas densidades no microhabitat ciliar. Não houve registro de E. heterophylla no microhabitat ciliar. O tamanho das populações, as taxas de mortalidade e de natalidade variaram ao longo do tempo em cada microhabitat. Este estudo indica que diferentes as condições de estabelecimento influenciaram o tamanho das populações e a ocorrência das quatro espécies e confirma que microhabitat pode atenuar o efeito do estresse hídrico sobre a mortalidade durante a estação seca, mas a força desse papel atenuador pode variar com o tempo e a espécie considerada.

Chronic hepatitis B and liver schistosomiasis: a deleterious association.

BACKGROUND: Chronic hepatitis B (CHB) and schistosomiasis are prevalent in several countries, but the impact of this association is unknown. We aimed to investigate the prevalence and morbidity of this co-infection in Minas Gerais, an endemic area of schistosomiasis in Brazil. METHODS: In total, 406 adults with CHB (HBsAg positive >6 months) were included in a cross-sectional study. CHB was classified as replicative (HBV DNA ≥ 2.000 IU/ml), and low replicative or inactive hepatitis B carriers (HBV DNA <2.000 IU/ml). Schistosomiasis was confirmed by epidemiological and clinical records. Liver biopsies were scored by METAVIR. The risk of severe fibrosis was estimated by multivariate analysis. RESULTS: Of the 406 patients, 64.8% (263) were male, and the median age was 45 years (IQR 35-54). In total, 57.9% (235) had replicative CHB, and 31.5% (128) had cirrhosis. Schistosoma mansoni was confirmed in 30.5% (124) patients, 81.5% (101) of which were male with a median age of 47 years (IQR 39.5-54). Of the co-infected patients, 61.3% (76) and 38.7% (48) had replicative and inactive CHB, respectively. Schistosomal portal fibrosis (PF) was detected in 69.4% (86/124) patients. Patients with replicative CHB and schistosomal PF had more advanced fibrosis and severe inflammation compared with patients without schistosomal PF (80.8% vs 43.6% for METAVIR F3-F4, p<0.01; 64.0% vs 39.8% for METAVIR A2-A3, p < 0.01). Age >50 years (OR = 1.10; 95% CI 1.06-1.14, p<0.001), male gender (OR = 2.61, 95% CI 1.12-6.09, p = 0.03), schistosomal PF (OR = 4.56, 95% CI 2.10-9.91, p<0.001) and alcoholism (OR = 2.46, 95% CI 1.16-5.19, p = 0.02) were independently associated with cirrhosis. CONCLUSIONS: The association between replicative CHB and schistosomal PF can be a risk factor for more severe liver disease, which can result in deleterious outcomes for patients from endemic areas.

Genetic relatedness of a non-motile variant O157 enteropathogenic Escherichia coli (EPEC) strain and E. coli strains belonging to pathogenic related groups.

The study was undertaken to determine the clonal relationship and the genetic diversity among Escherichia coli isolates by comparing a non-motile O157 variant with three O157:H7 EHEC isolates and one O55:H7 enteropathogenic E. coli (EPEC) strain. E. coli strains were characterized by sorbitol phenotype, multilocus enzyme electrophoresis, pulsed-field gel electrophoresis, random amplification polymorphic DNA, and the presence of specific virulence genes (stx, E-hly and LEE genes). Sorbitol fermentation was observed in O157:H- (strain 116I), O55:H7 and O157:H7 (strain GC148) serotypes. stx1 or stx2 and E-hly genes were only detected among O157:H7 isolates. LEE typing revealed specific allele distribution: eaegamma, tirgamma, espAgamma, espBgamma associated with EPEC O55:H7 and EHEC O157:H7 strains (B1/1 and EDL 933), eaealpha, tiralpha, espAalpha, espBalpha related to the 116I O157:H- strain and the GC148 strain presented non-typable LEE sequences. Multilocus enzyme profiles revealed two main clusters associated with specific LEE pathotypes. E. coli strains were discriminated by random amplification of polymorphic DNA-polymerase chain reaction and pulsed-field gel electrophoresis methodologies. The molecular approaches used in this study allowed the determination of the genetic relatedness among E. coli strains as well as the detection of lineage specific group markers.

Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk.

Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.

Estudo epidemiológico de problemas reprodutivos em rebanhos bovinos na bacia leiteira de Goiânia / An epidemiological study of reproductive failure in dairy herds from Goiânia

No período de março 2001 a julho 2002 foram estudadas 2823 fêmeas bovinas, em idade de reprodução, mestiças das raças Holandesa e Gir, pertencentes a 34 propriedades da bacia leiteira de Goiânia. A taxa de prenhez foi 47,8%. Entre os animais não gestantes (n= 1473), 17% apresentaram alterações inflamatórias do útero. Outras anormalidades encontradas foram: aplasia parcial do sistema genital (0,04%), feto macerado (0,01%), ovário aderido (0,04%). Casos de natimortalidade (0,04%), retenção placentária (0,01 %) e cervicite (0,6%) e taxas de abortos (0,88%) também foram registrados. Cocos Gram positivos (41,3%) e bastonetes Gram negativos (52,6%) foram os microrganismos mais isolados sendo Staphylococcus aureus e Escherichia colli os principais patógenos encontrados nas infeccções uterinas. Testes in vitro mostraram que esses microrganismos apresentaram maior susceptibilidade frente ao c1oranfenicol, à gentamicina e à neomicina.

Outbreaks of cholera-like diarrhoea caused by enterotoxigenic Escherichia coli in the Brazilian Amazon Rainforest.

The relationship between enteropathogens and severe diarrhoea in the Brazilian Amazon is poorly understood. In 1998, outbreaks of acute diarrhoea clinically diagnosed as cholera occurred in two small villages localized far from the main cholera route in the Brazilian rainforest. PCR was performed on some enteropathogens and heat-labile (LT) and/or heat-stable (STh) toxin genes, the virulence determinants of enterotoxigenic Escherichia coli (ETEC), were detected. Further characterization of ETEC isolates revealed the presence of two clones, one from each outbreak. One presenting serotype O167:H5 harboured LT-I and STh toxin genes and expressed the CS5CS6 colonization factor. The other, a non-typeable serotype, was positive for the LT-I gene and expressed the CS7 colonization factor. The current study demonstrates the importance of molecular diagnosis in regions such as the Amazon basin, where the enormous distances and local support conditions make standard laboratory diagnosis difficult. Here we also show that the mis-identified cholera cases were in fact associated with ETEC strains. This is the first report of ETEC, molecularly characterized as the aetiological agent of severe diarrhoea in children and adults in the Brazilian Amazon Rainforest.

Frequency and characteristics of diarrhoeagenic Escherichia coli strains isolated from children with and without diarrhoea in Rio de Janeiro, Brazil.

The frequency of diarrhoeagenic Escherichia coli (DEC) strains was investigated in 253 children up to 3 years old, with (patient group, PG, 199 children) and without (control group, CG, 54 children) diarrhoea, living in Rio de Janeiro, Brazil. DEC strains were detected in 70 (27.6%) children, including 54 (27.1%) with diarrhoea and 16 (29.6%) without diarrhoea. Enteroaggregative E. coli (EAEC) was the most frequent DEC category, accounting for 14.6% of the isolates in the PG and for 11.1% in the CG. E. coli strains carrying enteropathogenic E. coli (EPEC) virulence markers showed higher incidence in the CG (12.9%) than in the PG (8.0%). E. coli strains belonging to non-classical EPEC groups that carried eae only or eae and bfpA, designated as attaching-effacing E. coli (AEEC) were the most frequent (79.1%). Simultaneous presence of multiple EPEC virulence factors (EAF/eae/bfpA) were only detected among strains isolated from the PG. Enterotoxigenic E. coli (ETEC) strains were isolated from 5.5% of the children in the CG and from 3.5% of those in the PG. Most of the ETEC isolates were LT-probe positive (70%) and none carried both LT-I and ST-I probe sequences. One enteroinvasive E. coli (EIEC) strain was recovered from a child with diarrhoea. No stx-probe positive E. coli strains were detected. Overall, DEC strains were not found to be significantly associated with diarrhoea (p>0.05). However, the higher incidence of EAEC, the most frequent DEC category, among children with diarrhoea, suggests a potential role of EAEC as an important enteric pathogen in the community investigated.

Serobiotypes and virulence genes of Escherichia coli strains isolated from diarrheic and healthy rabbits in Brazil.

A total of 178 Escherichia coli isolates from diarrheic and healthy rabbits in the São Paulo State (Brazil) were serobiotyped and investigated by PCR for the presence of virulence genes. Among the 90 (50.6%) isolates which possessed the eae gene, 74 were from diarrheic animals and all but one encoded intimin beta. Sixty five (72.2%) of the eae+ isolates had insertion of the locus of enterocyte effacement locus in the pheU locus, 11 (12.2%) in the selC and 14 (15.6%) did not insert in either of these loci. All isolates were negative for genes of the E. coli enterotoxins, Stx1, Stx2, CNF1, CNF2 and EHEC hemolysin. The O132:H2 serotype was dominant, being present in 63 isolates (70%) of the 90 eae+ isolates, and 57 of the 63 isolates of this serotype belonged to biotype 30. PCR detected the gene for AF/R2 fimbriae in 75 (83.3%) of the 90 eae+ isolates. Adherence to HeLa cells was best detected following 6h incubation and a positive fluorescence actin staining (FAS) test was given by 52 isolates. These data show that isolates of E. coli associated with diarrhea in rabbits in Brazil possess the genotype and phenotype typically associated with rabbit enteropathogenic E. coli (EPEC). We conclude that EPEC that possess the eae gene are a common cause of diarrhea in Brazilian rabbit farms and that the pathogenic eae+ AF/R2+ isolates of O132:H2:B30 serobiotype are especially predominant.

Bacteroides fragilis adherence to Caco-2 cells.

The ability of ten Bacteroides fragilis strains isolated from intestinal and non-intestinal infections, normal flora and environment to adhere to human colon carcinoma cells, Caco-2, was examined. The adherence capacity varied among the strains tested from strongly adherent (76-100%) to non- or weakly adherent (0-25%). Negative staining with Indian ink showed that all the strains were capsulated, although strain 1032 (strongly adherent and originated from bacteremia) had the highest rate of capsulated cells in the culture. All strains studied presented an electron-dense layer and no fimbrial structures in their surface after PTA negative staining. The analysis of the strains with ruthenium red showed the presence of an acidic polysaccharide and also surface vesicles in all of them. The strain 1032 presented an aggregative adherence pattern toward Caco-2 cells monolayers. It could be seen trapped by elongated microvilli and surrounded by extracellular material in the scanning electron microscope. Treatment with sodium periodate (100 mM/1 h) reduced significantly its adherence capacity and also the expression of an electron-dense layer and of the capsule, detected with PTA and Indian ink staining, respectively. We suggest that the capsular polysaccharide might mediate the adherence of the B. fragilis to Caco-2 cells.

Interactions of Escherichia coli strains of non-EPEC serogroups that carry eae and lack the EAF and stx gene sequences with undifferentiated and differentiated intestinal human Caco-2 cells.

Escherichia coli strains of non-EPEC serotypes that carry eae and lack the EAF and the Shiga toxin (stx) gene sequences have been found in acute diarrhea. Both the cell association and the cell entry of these strains in human intestinal epithelial cells were studied as a function of cell differentiation and polarization. The eae+/EAF-/stx- non-EPEC E. coli strains invaded undifferentiated Caco-2 cells more efficiently than differentiated cells. In contrast, prototype EPEC strain E2348/69 did not show significative differences from invasion rates of undifferentiated and differentiated cells. The uptake of these strains was greatly enhanced by pretreatment of differentiated Caco-2 cells with EGTA. These results suggest that the eae+/EAF-/stx- non-EPEC E. coli invasion of intestinal cells may be dependent on receptors expressed on the surface of undifferentiated cells and the basolateral pole of differentiated cells.

eae-negative attaching and effacing Escherichia coli from piglets with diarrhea.

One hundred and ninety strains of Escherichia coli that were isolated from pigs with diarrhea in the state of São Paulo, Brazil, and that were negative for enterotoxins and cytotoxins were investigated. Strains which adhered to HeLa cells were examined for fluorescence actin staining (FAS), the ability to induce attaching and effacing (A/E) lesions on HEp-2 cells detectable by transmission electron microscopy and the presence of eae gene sequences detected by PCR. Intimin production was detected by western blot and serogrouping was performed. Forty-seven isolates adhered to HeLa cells in several patterns, but none adhered in a localized adherence pattern. However, seven of the 47 adherent strains were positive for the FAS reaction, although the reactions were usually weak or atypical. One FAS-negative and three FAS-positive strains, which were examined for their ability to induce A/E lesions, were all positive. Subsequently, testing of these strains for the eae gene showed that they all lacked this gene. These findings, along with earlier reports of eae-negative A/E E. coli, suggest that higher quantities of E. coli in this category might be detected if more reliance were placed on phenotypic tests rather than on gene detection tests alone.

Phenotypic and genotypic characteristics of Escherichia coli strains of non-enteropathogenic E. coli (EPEC) serogroups that carry EAE and lack the EPEC adherence factor and Shiga toxin DNA probe sequences.

This study was conducted to characterize the virulence potential of 59 Escherichia coli strains carrying EAE and lacking the enteropathogenic E. coli adherence factor and Shiga toxin probe sequences. In hybridization studies, all strains carried the locus of enterocyte effacement (LEE)-associated DNA sequences. Of the other 15 virulence DNA sequences tested, HLY was the most frequent (44.1%); 17 combinations of these sequences were found, but strains carrying EAE only (EAE profile) were the most frequent (35.6%). Except for 1 cytodetaching strain, all others adhered to HeLa and Caco-2 cells, most of which (approximately 75.0%) showed variations of the localized adherence pattern. Actin accumulation was detected in 75.9% of the nondetaching strains. Most strains had LEE, probably inserted in pheU (49.2%), and presented a nontypeable intimin (83.1%). Translocated intimin receptor-derived DNA sequences correlated with enteropathogenic and enterohemorrhagic E. coli in 61.0% and 32.0% of the strains, respectively. Thirty-five different serotypes were found. Only strains with the EAE profile were associated with diarrhea (P=.039).

Structural response and relative strength of a laminated composite hip prosthesis: effects of functional activity.

To obtain a better appreciation for the structural performance of a laminated composite hip prosthesis (CP), we examined in situ prosthesis structural response and relative strengths as a function of walking and stair climbing using our previously developed analysis guidelines. Accordingly, we examined overall prosthesis structural response utilizing a global continuum level modeling approach and prosthesis relative strengths using a local microstructural (or ply-level) modeling approach. As a reference and control, we examined the structural performance of the intact natural femur (NAT) and a titanium alloy (Ti) based hip prosthesis. In terms of the overall structural response, i.e., the femur/prosthesis deformational response, stem/bone interfacial stress transfer, and calcar strain energy density restored, the performance of the CP prosthesis was moderately improved over that of the control Ti prosthesis and better approximates the NAT response. In terms of relative strength, we found that the neck of the CP prosthesis failed for all activities with the exception of the mid-stance phase of level walking. However, the prosthesis appears to have sufficient relative strength for function at positions distal to the neck of the prosthesis. While these results dampen enthusiasm for consideration of laminated composite hip prostheses designed with a shape based on a metal alloy implant, they indirectly support consideration of alternate hip prosthesis structural designs such as using a better supported prosthesis neck or utilizing metal/composite hybrid constructions. Importantly, our simulation and analysis approach could be utilized in the design of other laminated composite biomedical structural components.

In vitro effects of sildenafil and phentolamine, drugs used for erectile dysfunction, on human sperm motility.

OBJECTIVE: The aim of this study was to determine the effects of sildenafil and phentolamine on sperm motility in vitro. STUDY DESIGN: Semen or washed sperm was mixed with various doses of sildenafil or phentolamine and analyzed for motility during a 30-minute period. The pH was measured for each of the samples tested. Statistical analyses were performed with analysis of variance. RESULTS: A 200-microg/mL dose of sildenafil had no effect on sperm motility. However, the highest dose (2000 microg/mL) significantly reduced motility by about 50%. The pH was reduced in this high-dose sample. The lowest dose of phentolamine (20 microg/mL) had no effect, whereas a dose of 200 microg/mL resulted in a significant reduction in sperm motility. The highest dose (2000 microg/mL) stopped virtually all sperm from moving. CONCLUSION: This study demonstrated a direct dose-related effect of phentolamine on reducing sperm motility. Only the highest dose of sildenafil had an effect, which may have been caused by a decline in pH.

Cytotoxic cell vacuolating activity from Vibrio cholerae hemolysin.

A Vibrio cholerae cytotoxin, designated VcVac, was found to cause vacuolation in Vero cells. It was originally detected in the pathogenic O1 Amazonia variant of V. cholerae and later shown to be produced in environmental strains and some El Tor strains. Comparison of VcVac production in various strains suggested that hemolysin was responsible for the vacuolating phenotype. Genetic experiments established a firm correlation between vacuolation and hemolysin production. The mammalian cell vacuolating activity of the V. cholerae hemolysin is a new property of this protein and points to a previously unknown type of interaction between V. cholerae and its host.

High occurrence of Shiga toxin-producing Escherichia coli (STEC) in healthy cattle in Rio de Janeiro State, Brazil.

In order to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) strains, 197 fecal samples of healthy cattle from 10 dairy farms, four beef farms and one slaughterhouse at Rio de Janeiro State, Brazil, were examined for Shiga toxin (Stx) gene sequences by polymerase chain reaction (PCR). For presumptive isolation of O157:H7 E. coli, the Cefixime-potassium tellurite-sorbitol MacConkey Agar (CT-SMAC) was used. A high occurrence (71%) of Stx was detected, and was more frequently found among dairy cattle (82% vs. 53% in beef cattle), in which no differences were observed regarding the age of the animals. Dot blot hybridization with stx1 and stx2 probes revealed that the predominant STEC type was one that had the genes for both stx1 and stx2 in dairy cattle and one that had only the stx1 gene for beef cattle. Three (1.5%) O157:H7 E. coli strains were isolated from one beef and two dairy animals by the use of CT-SMAC. To our knowledge, this is the first report of O157:H7 isolation in Brazil. A PCR-based STEC detection protocol led to the isolation of STEC in 12 of 16 randomly selected PCR-positive stool samples. A total of 15 STEC strains belonging to 11 serotypes were isolated, and most of them (60%) had both stx1 and stx2 gene sequences. Cytotoxicity assays with HeLa and Vero cells revealed that all strains except two of serotype O157:H7 expressed Stx. The data point to the high prevalence of STEC in our environment and suggest the need for good control strategies for the prevention of contamination of animal products.

Integrated analysis of thrombotic/vascular event occurrence in epoetin alfa-treated patients undergoing major, elective orthopedic surgery.

Data from four prospective, multicenter, randomized studies involving 869 major, elective orthopedic surgery patients were examined by means of a retrospective integrated analysis to evaluate whether perioperative Epoetin alfa use was associated with the occurrence of thrombotic/vascular events. The incidence of thrombotic/vascular events was similar between 619 patients treated with Epoetin alfa and 250 patients receiving placebo (7.4% versus 8.0%, respectively). Regression analyses identified age, cardiac history, hypertension, and cardiac medications, but not Epoetin alfa, as risk factors for thrombotic/vascular events. The analysis did not implicate an increase in the rate of rise in hematocrit or maximum hematocrit obtained prior to surgery as contributors to thrombotic events. Thus, Epoetin alfa, which enhances preoperative erythropoiesis and increases hematocrit, did not affect the probability of thrombotic/vascular events.