A Weight of Evidence approach to support the assessment of the quality of Manila clam farming sites in a coastal lagoon.

Aquaculture productivity in coastal lagoons is endangered by a complex interplay of anthropogenic and environmental factors, amplified by the effects of climate change in these sensitive areas. To reach a more comprehensive assessment of farming sites quality, a quantitative Weight of Evidence approach (QWoE) is applied for the first time to data collected at four Manila clam (R. philippinarum) farming sites in the Venice lagoon (Italy). This included sediment quality, chemical bioaccumulation, and biological responses. Results revealed a greater hazard for sites closer to the open sea. In these areas, the combination of sediment characteristics and a higher frequency of salinity and temperature stress could explain the alterations measured at a transcriptional and biomarker level. The findings demonstrate that a QWoE approach that integrates multiple sources of evidence should also include physicochemical conditions in order to better understand the impacts of human activities and other stressors on clam aquaculture productivity.

Effects of environmental concentrations of the fragrance amyl salicylate on the mediterranean mussel Mytilus galloprovincialis.

Amyl salicylate (AS) is a fragrance massively used as a personal care product and following the discharged in wastewaters may end up in the aquatic environment representing a potential threat for the ecosystem and living organisms. AS was recently detected in water of the Venice Lagoon, a vulnerable area continuously subjected to the income of anthropogenic chemicals. The lagoon is a relevant area for mollusc farming, including the Mediterranean mussels (Mytilus galloprovincialis) having an important economic and ecological role. Despite high levels of AS occurred in water of the Lagoon of Venice, no studies investigated the possible consequences of AS exposures on species inhabiting this ecosystem to date. For the first time, we applied a multidisciplinary approach to investigate the potential effects of the fragrance AS on Mediterranean mussels. To reach such a goal, bioaccumulation, cellular, biochemical, and molecular analyses (RNA-seq and microbiota characterization) were measured in mussels treated for 7 and 14 days with different AS Venice lagoon environmental levels (0.1 and 0.5 µg L-1). Despite chemical investigations suggested low AS bioaccumulation capability, cellular and molecular analyses highlighted the disruption of several key cellular processes after the prolonged exposures to the high AS concentration. Among them, potential immunotoxicity and changes in transcriptional regulation of pathways involved in energy metabolism, stress response, apoptosis and cell death regulations have been observed. Conversely, exposure to the low AS concentration demonstrated weak transcriptional changes and transient increased representation of opportunistic pathogens, as Arcobacter genus and Vibrio aestuarianus. Summarizing, this study provides the first overview on the effects of AS on one of the most widely farmed mollusk species.

The effects of glyphosate and AMPA on the mediterranean mussel Mytilus galloprovincialis and its microbiota.

Glyphosate, the most widely used herbicide worldwide, targets the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme in the shikimate pathway found in plants and some microorganisms. While the potential for glyphosate to induce a broad range of biological effects in exposed organisms has been demonstrated, the global molecular mechanisms of toxicity and potential effects in bacterial symbionts remain unclear, in particular for ecologically important marine species such as bivalve molluscs. Here, the effects of glyphosate (GLY), its degradation product aminomethylphosphonic acid (AMPA), and a mixture of both (MIX) on the mussel M. galloprovincialis were assessed in a controlled experiment. For the first time, next generation sequencing (RNA-seq and 16S rRNA amplicon sequencing) was used to evaluate such effects at the molecular level in both the host and its respective microbiota. The results suggest that the variable capacity of bacterial species to proliferate in the presence of these compounds and the impairment of host physiological homeostasis due to AMPA and GLY toxicity may cause significant perturbations to the digestive gland microbiota, as well as elicit the spread of potential opportunistic pathogens such as Vibrio spp.. The consequent host-immune system activation identified at the molecular and cellular level could be aimed at controlling changes occurring in the composition of symbiotic microbial communities. Overall, our data raise further concerns about the potential adverse effects of glyphosate and AMPA in marine species, suggesting that both the effects of direct toxicity and the ensuing changes occurring in the host-microbial community must be taken into consideration to determine the overall ecotoxicological hazard of these compounds.

Ecotoxicological effects of the herbicide glyphosate in non-target aquatic species: Transcriptional responses in the mussel Mytilus galloprovincialis.

Glyphosate has been the most widely used herbicide worldwide over the last three decades, raising increasing concerns for its potential impacts on environmental and human health. Recent studies revealed that glyphosate occurs in soil, surface water, and groundwater, and residues are found at all levels of the food chain, such as drinking water, plants, animals, and even in humans. While research has demonstrated that glyphosate can induce a broad range of biological effects in exposed organisms, the global molecular mechanisms of action still need to be elucidated, in particular for marine species. In this study, we characterized for the first time the molecular mechanisms of action of glyphosate in a marine bivalve species after exposure to environmentally realistic concentrations. To reach such a goal, Mediterranean mussels Mytilus galloprovincialis, an ecologically and economically relevant species, were exposed for 21 days to 10, 100, and 1000 µg/L and digestive gland transcriptional profiles were investigated through RNA-seq. Differential expression analysis identified a total of 111, 124, and 211 differentially regulated transcripts at glyphosate concentrations of 10, 100, and 1000 µg/L, respectively. Five genes were found consistently differentially expressed at all investigated concentrations, including SERP2, which plays a role in the protection of unfolded target proteins against degradation, the antiapoptotic protein GIMAP5, and MTMR14, which is involved in macroautophagy. Functional analysis of differentially expressed genes reveals the disruption of several key biological processes, such as energy metabolism and Ca2+ homeostasis, cell signalling, and endoplasmic reticulum stress response. Together, the results obtained suggest that the presence of glyphosate in the marine ecosystem should raise particular concern because of its significant effects even at the lowest concentration.

Clam bioaccumulation of Alkylphenols and Polyciclic aromatic hydrocarbons in the Venice lagoon under different pressures.

Biota-Sediment Accumulation Factors (BSAFs) of nonylphenols (NPs) and polycyclic aromatic hydrocarbons (PAHs) in Ruditapes philippinarum from the Venice Lagoon (Italy) were determined with the aim to verify whether the routine biomonitoring studies are reliable in contaminated sites. Clams and sediments were collected in field campaigns (October 2003 to June 2004) in three sites of the Venice Lagoon. Results showed that Marghera and Campalto sediments were more contaminated by NPs and PAHs than Poveglia. Different trends were observed in the contamination of clams with the highest BSAFs found at Poveglia. BSAF trend appeared to be inversely related to the contaminant pressure on the sites. These results suggest that clam bioaccumulation is not always representative of the chemical pressure on aquatic biota. The direct correlation between sediment and biota concentrations in contaminated sites can be lost as a function of the site-specific conditions such as sediment toxicity and food availability.

Does seawater acidification affect survival, growth and shell integrity in bivalve juveniles?

Anthropogenic emissions of carbon dioxide are leading to decreases in pH and changes in the carbonate chemistry of seawater. Ocean acidification may negatively affect the ability of marine organisms to produce calcareous structures while also influencing their physiological responses and growth. The aim of this study was to evaluate the effects of reduced pH on the survival, growth and shell integrity of juveniles of two marine bivalves from the Northern Adriatic sea: the Mediterranean mussel Mytilus galloprovincialis and the striped venus clam Chamelea gallina. An outdoor flow-through plant was set up and two pH levels (natural seawater pH as a control, pH 7.4 as the treatment) were tested in long-term experiments. Mortality was low throughout the first experiment for both mussels and clams, but a significant increase, which was sensibly higher in clams, was observed at the end of the experiment (6 months). Significant decreases in the live weight (-26%) and, surprisingly, in the shell length (-5%) were observed in treated clams, but not in mussels. In the controls of both species, no shell damage was ever recorded; in the treated mussels and clams, damage proceeded via different modes and to different extents. The severity of shell injuries was maximal in the mussels after just 3 months of exposure to a reduced pH, whereas it progressively increased in clams until the end of the experiment. In shells of both species, the damaged area increased throughout the experiment, peaking at 35% in mussels and 11% in clams. The shell thickness of the treated and control animals significantly decreased after 3 months in clams and after 6 months in mussels. In the second experiment (3 months), only juvenile mussels were exposed to a reduced pH. After 3 months, the mussels at a natural pH level or pH 7.4 did not differ in their survival, shell length or live weight. Conversely, shell damage was clearly visible in the treated mussels from the 1st month onward. Monitoring the chemistry of seawater carbonates always showed aragonite undersaturation at 7.4 pH, whereas calcite undersaturation occurred in only 37% of the measurements. The present study highlighted the contrasting effects of acidification in two bivalve species living in the same region, although not exactly in the same habitat.

First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea, Decapoda).

For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3-5 microm in diameter and 30-50 fL in volume, the second was of about 6-12 microm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20+/-1.7 microm and 272.30+/-143.5 fL, respectively. Haemocytes observed under light microscope were distinguished in three cell types: granulocytes (28%; 11.94+/-1.43 microm in diameter) with evident cytoplasmic granules, semigranulocytes (27%; 12.38+/-1.76 microm in diameter) with less granules than granulocytes, and hyalinocytes (44%; 7.88+/-1.6 microm in diameter) without granules. In addition, a peculiar cell type was occasionally found (about 1%): it was 25-30 microm in diameter and had a great vacuole and a peripheral cytoplasm with granules. Granulocyte and semigranulocyte granules stained in vivo with Neutral Red, indicating that they were lysosomes. Giemsa's dye confirmed that granulocytes and semigranulocytes were larger than hyalinocytes. Pappenheim's panoptical staining and Ehrlich's triacid mixture allowed to distinguish granule-containing cells (including semigranulocytes) in acidophils (64%), basophils (35%) and neutrophils (1%). Hyalinocytes showed always a basophilic cytoplasm. Haemocytes were positive to the PAS reaction for carbohydrates, even if cytoplasm carbohydrate distribution varied among cell types. Lastly, lipids were found on cell membrane and in cytoplasm of all haemocyte types in the form of black spots produced after Sudan Black B staining. The morphological characterisation of C. aestuarii haemocytes by light microscopy was necessary before performing both ultrastructural and functional studies of circulating cells.

Investigation of EROD, CYP1A immunopositive proteins and SOD in haemocytes of Chamelea gallina and their role in response to B[a]P.

CYP1A sub-family represents the main form of cytochrome P450 involved in benzo[a]pyrene (B[a]P) detoxification, but there are no clear evidences about its presence in invertebrates. 7-Ethoxy resorufin O-deethylase (EROD) activity is strictly related to CYP1A presence, at the same time P450-dependent oxidative metabolism leads to reactive oxygen species (ROS) production, thought to be an important mechanism of pollutant-mediated toxicity in aquatic organisms. Superoxide dismutases (SODs), EROD and CYP1A activities and/or expressions were detected in haemocytes of pooled clams (Chamelea gallina) and cell-free haemolymph after 24 h, 7 and 12 days of exposure to 0.5 mg/L of B[a]P. After 24 h, B[a]P content was maximum in whole tissues. A 61 kDa band was recognized in haemocytes and cell-free haemolymph by polyclonal anti-fish CYP1A, while 53.5 and 63.8 kDa CYP1A immunopositive proteins were discriminate without differences of expression. Differently, EROD, MnSOD activity/expression and ECSOD expression decreased in haemocytes and haemolymph. C. gallina immune system presents an interesting response dose/time exposure of B[a]P and the 7 days condition highlights the major effects of xenobiotic action. The identification of basal EROD levels supports the possible presence of the CYP1A, never identified in C. gallina and more specifically never isolated in immune cells, as confirmed by CYP1A-immunopositive proteins identification.

Uptake and elimination of 4-nonylphenol by the clam Tapes philippinarum.

The present study is the first dealing with the accumulation and elimination of 4-nonylphenol (NP) by the Manila clam, Tapes philippinarum. Specimens of T. philippinarum were exposed to NP-spiked seawater, and the NP contents in whole soft tissue, gills, digestive gland, and haemolymph were measured. Actual NP concentrations in seawater and microalgae (Isochrysis galbana) used for feeding were also determined, and the algal bioconcentration factor (BCF) value (640 ml/g) was calculated. Volatilisation was the main cause of dissipation of NP from experimental tanks, attaining up to 78% of the NP added. NP accumulated by algae used for feeding was negligible with respect to the total added NP, and we concluded that in our experiments, clams accumulated NP mainly from water and not food intake. Accumulation followed a two-compartment, first-order rate coefficient model, with an uptake rate coefficient of 13.8 +/- 0.6 mL g(-1)h(-1)(fresh weight [fw]) and an elimination coefficient of 0.0070 +/- 0.0005 h(-1). Ninety percent of the steady state was reached after 14 days of exposure, and the BCF value at the steady state was 1958 +/- 158 mL g(-1) fw (1.8 +/- 0.2 x 10(5) based on lipid weight). Slightly less than 50% of NP bioaccumulated through water was allocated into the gills, whereas the rest was found in the digestive gland. In the first 8 hours, clams eliminated 51% of the NP accumulated, and only 2% of the NP accumulated was detected in the clams at the end of the elimination phase (day 14). Two-compartment first-order decay model described the elimination of the accumulated NP by considering the clam as two compartments each with a different elimination rate. The sudden elimination of NP in the initial hours can be attributed to the elimination of NP accumulated into the gills and readily available for excretion (k ( e ) = 0.30 +/- 0.07 h(-1)). The slower step of the elimination process should be the mobilisation of NP accumulated in internal organs, which must be carried into the haemolymph for excretion (k(e) = 0.0091 +/- 0.0002 h(-1)). Because T. philippinarum has been demonstrated to accumulate NP dissolved in water, high NP levels can be hypothesised in clams from highly contaminated environments. This research was performed according to all national and international guidelines for animal welfare.

Field validation of autometallographical black silver deposit (BSD) extent in three bivalve species from the Lagoon of Venice, Italy (Mytilus galloprovincialis, Tapes philippinarum, Scapharca inaequivalvis) for metal bioavailability assessment.

The autometallographic approach was applied to three filter-feeder bivalves from differently polluted sites of the Lagoon of Venice. The area density (AD) values of black silver deposits (BSDs) were therefore seasonally quantified by image analysis in digestive cell lysosomes. Sediments were characterised in terms of grain size, organic content and metal concentrations (Cd, Cu, Ni, Pb, Zn). The physiological status of the bivalves was also evaluated by applying the "survival in air" test as a generic stress index. Chemical results showed that Zn contributed to the total metal load for more than 60% in sediments and for 85%-94% in mussel and clam digestive glands, respectively (data processed from published results obtained on the same samples). Regression analyses between extent of BSDs and both total metal and Zn concentrations in digestive gland followed a linear pattern in mussels and clams, but not in arks. Similarly, in mussels and clams only, BSD amount was significantly correlated with total metal concentrations in sediments according to a logarithmic model. Consequently, results obtained from autometallography can discriminate sites on the basis of their metal pollution levels, like chemical analyses on both sediments and digestive glands, evaluations being supported by the results of the physiological stress index. As a conclusion, it is suggested that autometallography in bivalves may provide overall estimates of metal fractions moving from sediments towards organisms through the ingestion of metal-enriched particles. Future work is needed to define the bioaccumulation model in arks and to clarify their possible use as metal biomonitors.

Effects of copper and cadmium exposure on functional responses of hemocytes in the clam, Tapes philippinarum.

The effects of Cu and Cd on functional responses of hemocytes in Tapes philippinarum, a Veneridae clam widespread in the lagoon of Venice, were evaluated. Bivalves were exposed for 7 days to various concentrations of CuCl(2) x 2H(2)O (0, 10, 60, and 110 microg/L) and CdCl(2) x H(2)O (0, 150, 300, and 450 microg/L). Hemocytes were collected from exposed clams, and the effects of Cu and Cd on phagocytosis, Neutral Red retention capacity, superoxide dismutase, and cytochrome oxidase activities were investigated. Hemocytes from animals exposed to Cu showed a significant decrease in phagocytic activity, whereas no inhibition was observed in cells from Cd-exposed animals. Diffusion of Neutral Red into the cytosol, an index of lysosomal membrane alteration, was enhanced by high concentrations of both metals. Different results in biochemical parameters were observed. Exposure to 60 and 110 microg Cu/L caused a significant reduction in hemocyte superoxide dismutase activity, whereas no changes resulted after Cd exposure. Increased cytochrome oxidase activity was observed in hemocytes from mollusks exposed to 60 microg Cu/L and 300 microg Cd/L; no significant differences were found in cells from bivalves exposed to 110 microg Cu/L and 450 microg Cd/L. These results suggest a relationship between heavy metal exposure and alterations in functional responses of hemocytes in T. philippinarum and indicate that the type of observed effects vary with the nature and concentration of heavy metals. Moreover, the data obtained in the analyzed clam support the possibility of using it as sentinel organism in biomonitoring studies, even if used biomarkers will be further evaluated in field conditions.

Prevention of nosocomial bloodstream infections: effectiveness of antimicrobial-impregnated and heparin-bonded central venous catheters.

OBJECTIVES: To examine the effectiveness of antimicrobial-impregnated and heparin-bonded catheters relative to standard central venous catheters in lessening catheter-related bloodstream infections. DATA SOURCES: Articles were identified by computer-assisted searching. STUDY SELECTION: Studies were eliminated from further consideration if they did not contain original data relevant to lessening catheter-related bloodstream infections, were nonrandomized or uncontrolled, described subjects <17 yrs of age, or used animal subjects. DATA ABSTRACTION: From each eligible article, we abstracted the following: a) citation; b) type of control; c) study setting; d) type of experimental catheter; e) catheter-specific complications; f) total numbers of patients and catheters; g) number of experimental catheters used that resulted in a catheter-related bloodstream infection; h) number of control catheters used that resulted in a catheter-related bloodstream infection; i) number of experimental catheters used without catheter-related bloodstream infections; and j) number of control catheters used without infections. We also recorded the duration of catheter use and the types of microbes cultured in association with the catheters and with catheter-related bloodstream infections. DATA SYNTHESIS: Eleven eligible studies were identified. Using meta-analysis, we showed that antimicrobial-impregnated and heparin-bonded central venous catheters significantly decreased catheter-related bloodstream infections by 2.32% (95% confidence interval, 1.04% to 3.61%). CONCLUSIONS: The modest additional cost for the use of these catheters relative to the considerable cost of treating even a single bloodstream infection makes their use cost-effective.

Adverse behavioral and sexual factors in chronic vulvar disease.

OBJECTIVE: We investigated the prevalence of adverse sexual, personal hygiene, and self-treatment behavior among women with chronic benign vulvar and vaginal disorders. STUDY DESIGN: This study was carried out by means of a scientific clinical audit producing actuarial data by way of a semistructured questionnaire. Subjects consisted of a sample of 530 cases treated at a specialist clinic for vulvar diseases. In addition to clinical and laboratory investigations, the patients were interviewed by the clinic's psychologist, who used a structured interview schedule developed specifically for the study. Descriptive analysis and analysis of variance were used to investigate data. RESULTS: Most patients remained sexually active in spite of arousal failure and discomfort and engaged in unlubricated and often painful sexual intercourse. Most women engaged in potentially harmful genital hygiene and self-treatment behavior. Adverse self-treatment practices but not adverse genital hygiene practices were associated with greater symptom duration. CONCLUSIONS: Adverse practices should be addressed during the management of cases of chronic vulvar disease.

Haemocytes of the clam Tapes philippinarum (Adams & Reeve, 1850): morphofunctional characterisation.

Tapes philippinarum is a bivalve mollusc of the Pacific Ocean, successfully imported for human consumption into the northern Adriatic Sea (Europe). For better knowledge of its considerable adaptive ability in comparison with similar autochthonous species, a morpho-functional characterisation of its haemocytes was carried out with the establishment of short-term cell cultures (60 min at 25 degrees C). Various methods of cytochemical staining identified four cell types in the haemolymph: granulocytes (48.05% +/- 1.43), hyalinocytes (32.18% +/- 0.99), haemoblasts (18.97% +/- 0.63) and serous cells (0.8% +/- 0.19). The granulocytes, possessing cytoplasmic granules with differing dye affinity, included basophils, neutrophils and acidophils. Such granules stained vitally with Neutral Red, and correspond to lysosomes. Hydrolytic and oxidative enzymes were mainly detectable after stimulation in the presence of yeast cells. Both granulocytes and hyalinocytes were positive for alkaline phosphatase, non-specific esterase, peroxidase, and cytochrome C oxidase, whereas only granulocytes were positive for beta-glucuronidase, acid esterase, and arylsulphatase. Both cell types were competent phagocytes towards yeast and plasma had an opsonising effect. Moreover, the respiratory burst accompanied phagocytosis with superoxide anion production, recognisable through cytoplasmic deposits of formazan after treatment with nitro blue tetrazolium. Haemoblasts were small undifferentiated cells which, due to their morphology and positivity to the anti-CD34 antibody, show the typical features of stem cells. Serous cells, probably arising from Keber's gland and belonging to another differentiation pathway, contained non-sulphate acid mucopolysaccharides and play an important role in early defence mechanisms, taking part in the formation of clots.

Embryotoxicity of butyltin compounds to the sea urchin Paracentrotus lividus.

Tributyltin (TBT) has been widely employed in marine anti-fouling paints as a biocide, although it represents a serious risk, particularly in estuarine and coastal water/sediment ecosystems. In this study, the embryotoxic effects of TBT and its degradation products, dibutyltin (DBT) and monobutyltin (MBT), were analyzed during the development of the sea urchin Paracentrotus lividus from post-fertilization to the pluteus stage, to better clarify ecotoxicological impact. The embryotoxicity of butyltins is concentration-dependent and increases proportionally with number of butyl groups. Significant growth reduction was observed at TBT concentrations as low as 0.01 microgram l-1; 1 microgram l-1 was the maximum concentration allowing embryos to reach the pluteus stage at 48 h post-fertilization. Development was blocked at the morula or blastula stage with higher TBT concentrations. DBT and MBT are less toxic: slowed development and a decrease in pluteus size occurred at 10 micrograms l-1 DBT and 0.5 mg l-1 MBT. Effects on both skeletal deposition and blocked embryonic development are suggested to be due to the interference of organotin compounds with intracellular calcium homeostasis.

Cost-effectiveness of a post-exposure HIV chemoprophylaxis program for blood exposures in health care workers.

We performed a cost-effectiveness analysis of a post-exposure chemoprophylaxis program for health care workers who sustained exposures to blood. We analyzed a program of (1) treatment with zidovudine alone versus no treatment and (2) treatment with three-drug therapy versus no treatment. Assuming that 35% of exposures were to HIV-positive sources, the zidovudine regimen prevented 53 HIV seroconversions per 100,000 exposures, at a societal cost of $2.0 million per case of HIV prevented. The cost per quality-adjusted life year saved was $175,222. A three-drug chemoprophylactic therapy program (postulating 100% effectiveness and 35% source HIV positivity), prevented 66 seroconversions per 100,000 exposures, at a cost of $2.1 million per case of HIV prevented and $190,392 per quality-adjusted life year saved. Treating sources known to be HIV-positive and treating severe exposures were the most cost-effective strategies.

Immunotoxic effects of organotin compounds in Tapes philippinarum.

One of the most harmful groups of coastal pollutants is the organotin compounds (OTCs) which have severe effects on both aquatic organisms and mammals including humans. The immunotoxic effects of OTCs were studied in the cultivated clam Tapes philippinarum by determining the immunosuppressant role on in vitro yeast phagocytosis at low doses (0.01, 0.05, 0.1 microM). The phagocytic index was significantly reduced in an irreversible non-lethal manner depending on concentration and lipophilic affinity. The order of inhibition was TBT > or = DBT > MBT for butyltins and TPTC > TPTA > or = TPTH for triphenyltins.

Dyspareunia and vulvar disease.

OBJECTIVE: To determine the presence and frequency of contributing factors in functional dyspareunia and if these factors differ between patients with an epithelial disorder and those with apparently healthy genitalia. STUDY DESIGN: A psychologist assessed patients reporting dyspareunia at a vulvar diseases clinic by using a structured interview questionnaire, the Beck Depression Inventory, and the Spielberger State/Trait Anxiety Inventory. The questionnaire topics included sexual behavior, stresses and coping. Data were divided into two groups based on patients' functional diagnosis or epithelial disorder. RESULTS: Both groups recorded high levels of trauma-producing sexual behavior. Such behavior included unlubricated, unaroused and undesired sex. High levels of psychological and coping difficulties were also seen. Differences between the groups were seen in undesired sex, capacity to cope, stress factors coincidental with symptom onset and distress at the time of interview. CONCLUSION: Due to the high frequency of factors contributing to dyspareunia, such factors should be address regardless of the diagnosis. In all such patients it is recommended that a sexual and stress history be taken and counseling be considered.

Low-dose methotrexate spares steroid usage in steroid-dependent asthmatic patients: a meta-analysis.

STUDY OBJECTIVES: To determine if treatment with low-dose methotrexate spares oral steroids in adult, steroid-dependent, asthmatic patients. DESIGN: Articles identified by computer search were excluded if they (1) did not contain original data relating to the primary question, (2) had no controls, or (3) described subjects younger than 18 years of age. MEASUREMENTS: From each article, the following was abstracted: reference citation; type of control; whether study incorporated a run-in period in which the baseline level of prednisone was reduced to the lowest possible dose; dosage and length of methotrexate therapy; baseline dosage; and type of steroid. Also, the following was determined during both placebo and methotrexate arms of each study: steroid dosage, FEV1, serious side effects, and alteration in level of serum aspartate aminotransferase. RESULTS: Eleven eligible studies were identified. Methotrexate treatment resulted in a decrease in prednisone or prednisolone usage by an average of 4.37 mg/d or 23.7% of the initial dosage. The summary effect size in standard deviations (SDs) was -0.53 (95% confidence interval=-0.29 to -0.77). Subgroup analysis showed that patients treated with prolonged (6-month) therapy with methotrexate, those with low long-term usage of steroids (< or = 20 mg/d), and those whose study design incorporated a run-in period, tended to have the greatest steroid-sparing effects with methotrexate. CONCLUSIONS: Low-dose methotrexate has a significant steroid-sparing effect in steroid-dependent asthmatic patients. The greatest effect was evident in patients in whom an effort was made to reduce baseline steroid dosage and in whom methotrexate was used for 24 weeks.

Hepatitis C virus genotypes in northern Italy: clinical and virological features.

We analyzed the characteristics of subjects from the same area who were infected with hepatitis C virus genotypes 1 through 4 and subtypes 1a and 1b. Our data are consistent with a rapid evolution in the epidemiology of HCV genotypes and argue against different pathogenic potentials for genotypes 1b and 2.