A Rapid Field-Deployable Reverse Transcription-Insulated Isothermal Polymerase Chain Reaction Assay for Sensitive and Specific Detection of Bluetongue Virus.

Bluetongue is a non-contagious, haemorrhagic, Culicoides-borne disease of ruminants. The causative agent, bluetongue virus (BTV), is a member of the Orbivirus genus of the Reoviridae family. So far, 26 BTV serotypes have been identified worldwide. The global distribution of bluetongue has been expanding, and rapid detection of BTV, preferably in the field, is critical for timely implementation of animal movement restrictions and vector control measures. To date, many laboratory-based, molecular assays for detection of BTV have been developed. These methods require the samples to be shipped to a central laboratory with sophisticated instruments and highly skilled technicians to perform the assays, conduct analyses and interpret the results. Here, we report the development and evaluation of a rapid, portable, user-friendly, pan-BTV reverse transcription-insulated isothermal polymerase chain reaction (RT-iiPCR) assay that can potentially be used in low-resource field conditions. The total length of the assay was <60 min, and at the end of the assay, the results were automatically displayed as '+' or '-' without the need for data interpretation. The RT-iiPCR assay detected 36 BTV isolates and two in vitro transcribed RNA samples representing all 26 BTV serotypes. The assay did not cross-react with other animal viruses tested, including two closely related orbiviruses. The analytical sensitivity of the assay was as low as nine copies of in vitro transcribed double-stranded BTV RNA. Analysis of BTV-infected whole blood samples showed that the BTV RT-iiPCR assay was as sensitive as real-time RT-PCR. The assay can potentially be used for rapid screening of animals for BTV in routine diagnostics and for monitoring bluetongue outbreaks both in ruminants and in Culicoides vectors in the field and in the laboratory.

Incidence and outcomes of ventral hernia repair after robotic retropubic prostatectomy: A retrospective cohort of 570 consecutive cases.

BACKGROUND: Robotic retropubic prostatectomy (RRP) has become one of the most commonly performed robotic procedures in the United States. Ventral hernia (VH) has been increasingly recognized as an important complication after laparoscopic procedures, in general. However, data related to VH after robotic procedures is relatively scarce, especially after RRP. With increasing popularity of RRP, the purpose of this study was to look at the incidence of VH and outcomes of ventral hernia repair (VHR) after RRP. METHODS: All patients who underwent RRP at a single institution between January 2012 and June 2014 were studied retrospectively using electronic medical records. RESULTS: A total of 570 patients underwent RRP, of which 33 (5.8%) developed VH during the study period. Fourteen (42%) patients were obese and five (15%) had diabetes. One patient (3%) had a surgical site infection after RRP and two (6%) patients were on immunomodulators/steroids. Median duration to develop VH after RRP was 12 (1-25) months. Out of the 33 patients with VH, ten (33%) underwent VHR; five laparoscopic and five open. Median size of hernia defect and mesh used was 25 (1-144) cm2 and 181 (15-285) cm2, respectively. Median length of hospital stay and follow up was 0 (0-4) days and 12 (1-14) months, respectively. One patient who had initial VHR done at an outside institution had a recurrence. Thirty-two (97%) patients were alive at their last follow up. One patient died secondary to progression of prostate cancer. There was no significant 30 day morbidity (surgical site infection, fascial dehiscence, pneumonia, acute kidney injury, myocardial infarction). Of patients who decided non-operative management of VH (n = 23, 67%), none developed a complication requiring emergent surgical intervention. CONCLUSION: The incidence of VH after RRP is likely underreported in prior studies. Repair, either laparoscopic or open, is safe and effective in experienced hands. Patients who decide on watchful waiting can be followed with minimal risk of incarceration/strangulation. Further studies are needed to analyze the extraction techniques after RRP and correlate with incidence of VH.

Nomogram to predict perioperative blood transfusion for hepatopancreaticobiliary and colorectal surgery.

BACKGROUND: Predictive tools assessing risk of transfusion have not been evaluated extensively among patients undergoing complex gastrointestinal surgery. In this study preoperative variables associated with blood transfusion were incorporated into a nomogram to predict transfusion following hepatopancreaticobiliary (HPB) or colorectal surgery. METHODS: A nomogram to predict receipt of perioperative transfusion was developed using a cohort of patients who underwent HPB or colorectal surgery between January 2009 and December 2014. The discriminatory ability of the nomogram was tested using the area under the receiver operating characteristic (ROC) curve and internal validation performed via bootstrap resampling. RESULTS: Among 4961 patients undergoing either a HPB (56·3 per cent) or colorectal (43·7 per cent) resection, a total of 1549 received at least 1 unit of packed red blood cells, giving a perioperative transfusion rate of 31·2 per cent. On multivariable analysis, age 65 years and over (odds ratio (OR) 1·52), race (versus white: black, OR 1·58; Asian, OR 1·86), preoperative haemoglobin 8·0 g/dl or less (versus over 12·0 g/dl: OR 26·79), preoperative international normalized ratio more than 1·2 (OR 2·44), Charlson co-morbidity index score over 3 (OR 1·86) and procedure type (versus colonic surgery: major hepatectomy, OR 1·71; other pancreatectomy, OR 2·12; rectal surgery, OR 1·39; duodenopancreatectomy, OR 2·65) were associated with a significantly higher risk of transfusion and were included in the nomogram. A nomogram was constructed to predict transfusion using these seven variables. Discrimination and calibration of the nomogram revealed good predictive abilities (area under ROC curve 0·756). CONCLUSION: The nomogram predicted blood transfusion in major HPB and colorectal surgery.

The difficult gall bladder: Outcomes following laparoscopic cholecystectomy and the need for open conversion.

INTRODUCTION: Surgery for the difficult gallbladder (DGB) is associated with increased risk compared to more routine laparoscopic cholecystectomies (LC). Laparoscopic "damage control" methods including cholecystostomy, fundus-down approach and subtotal cholecystectomy (SC) have been proposed to avoid conversion to open. We hypothesized that a Total LC (TLC) for DBG can be completed safely with an acceptably low conversion rate. MATERIAL AND METHODS: All patients that underwent LC from January 2005-June 2015 were retrospectively reviewed. Cases met criteria for DGB if they were necrotic/gangrenous, involved Mirizzi syndrome, had extensive adhesions, were converted to open, lasted more than 120 min, had prior tube cholecystostomy or known GB perforation. RESULTS: A total of 2212 patients underwent LC during the study time period, of which 351 (15.8%) met criteria for DGB. Of these cases, 213 (60.7%) were admitted from the emergency department and 67 (19.1%) underwent urgent/emergent cholecystectomy (within 24 h). Additionally 18 (5.1%) had pre-operative tube cholecystostomies. Seventy patients (19.9%) were converted to open. Indications for conversion included severe inflammation/adhesion (n = 31, 46.3%), difficult anatomy (n = 14, 20.9%) and bleeding (n = 6, 9.0%). Predictors for conversion included urgent/emergent intervention (OR, 0.80; 95% CI 0.351-0.881, p = 0.032), previous abdominal surgery (OR, 2.18; 95% CI, 1.181-4.035, p = 0.013) and necrotic/gangrenous cholecystitis (OR, 1.92; 95% CI, 1.356-4.044, p = 0.033). Comparing the TLC and the conversion groups, mean operative time and length of hospital stay were significantly different; 147 ± 47 min vs 185 ± 71 min; p < 0.005 and 3 ± 2 days vs 5 ± 3 days; p = 0.011, respectively. There was no significant difference in postoperative hemorrhage, subhepatic collection, cystic duct leak, wound infection, reoperation and 30 day mortality. There was no bile duct injury in either group. CONCLUSION: Total laparoscopic cholecystectomy can be safely performed in difficult gallbladder situations with a lower conversion rate than previously reported. Possible predictors of conversion include urgency, necrotic gallbladder and history of prior abdominal surgeries. For patients converted to open, similar morbidity and mortality can be expected.

Development and validation of the Australian Aboriginal racial identity and self-esteem survey for 8-12 year old children (IRISE_C).

INTRODUCTION: In Australia, there is little empirical research of the racial identity of Indigenous children and youth as the majority of the current literature focuses on adults. Furthermore, there are no instruments developed with cultural appropriateness when exploring the identity and self-esteem of the Australian Aboriginal population, especially children. The IRISE_C (Racial Identity and Self-Esteem of children) inventory was developed to explore the elements of racial identity and self-esteem of urban, rural and regional Aboriginal children. This paper describes the development and validation of the IRISE_C instrument with over 250 Aboriginal children aged 8 to 12 years. METHODS: A pilot of the IRISE C instrument was combined with individual interviews and was undertaken with 35 urban Aboriginal children aged 8-12 years. An exploratory factor analysis was performed to refine the survey and reduce redundant items in readiness for the main study. In the main study, the IRISE C was employed to 229 Aboriginal children aged 6-13 years across three sites (rural, regional and urban) in Western Australia. An exploratory factor analysis using Principal axis factoring was used to assess the fit of items and survey structure. A confirmatory factor analysis was then employed using LISREL (diagonally weighted least squares) to assess factor structures across domains. Internal consistency and reliability of subscales were assessed using Cronbach's co-efficient alpha. RESULTS: The pilot testing identified two key concepts - children's knowledge of issues related to their racial identity, and the importance, or salience, that they attach to these issues. In the main study, factor analyses showed two clear factors relating to: Aboriginal culture and traditions; and a sense of belonging to an Aboriginal community. Principal Axis Factoring of the Knowledge items supported a 2-factor solution, which explained 38.7% of variance. Factor One (Aboriginal culture) had a Cronbach's alpha of 0.835; Factor 2 (racial identity) had a Cronbach's alpha of 0.800, thus demonstrating high internal reliability of the scales. CONCLUSION: The IRISE_C has been shown to be a valid instrument useful of exploring the development of racial identity of Australian Aboriginal children across the 8-12 year old age range and across urban, rural and regional geographical locations.

Challenging a surgical dictum: results from a 10-year experience on the safety of open inguinal herniorrhaphy in patients on chronic warfarin therapy.

BACKGROUND: Previous reports document the safety of open inguinal herniorrhaphy in patients on chronic warfarin therapy; however, the practice remains controversial. This study is a 10-year update of our experience. METHODS: A retrospective review of 1,839 consecutive patients undergoing open inguinal hernia repair was conducted from 2000 to 2010. All patients on chronic warfarin therapy were included. Three groups: continuation (CW), discontinuation (DW) and case-matched control (C) not on warfarin therapy were compared for operative details and postoperative complications. RESULTS: One hundred and fifty-eight patients were on chronic warfarin therapy. Of these, 40 patients (25%) continued on warfarin during the perioperative period (CW). Average preoperative international normalized ratio (INR) was 2.15 ± 0.76 for CW and 1.38 ± 0.42 for DW, p < 0.001. Mean operative times were equivalent between all three groups (88 min CW vs. 85 min DW vs. 79 min C, p = 0.518). Although CW patients experienced higher incidences of both hematoma and urinary retention overall, no statistically significant differences in complication rates were seen between the three groups (hematoma = 10 vs. 8% DW vs. 5% C, p = 0.703; urinary retention = 15 vs. 10% DW vs. 8% C, p = 0.541). Comparing patients by INR, there were no statistically different postoperative complication rates, particularly for hematoma (8% INR <2 vs. 9.5% INR = 2-3 vs. 20% INR >3, p = 0.65). CONCLUSION: Maintenance of warfarin therapy during the perioperative period for open inguinal herniorrhaphy results in equivalent operative times and postoperative complications as discontinuation.

Inguinal hernia repair in women: is the laparoscopic approach superior?

PURPOSE: Laparoscopic inguinal hernia repair is associated with reduced post-operative pain and earlier return to work in men. However, the role of laparoscopic hernia repair in women is not well reported. The aim of this study was to review the outcomes of the laparoscopic versus open repair of inguinal hernias in women and to discuss patients' considerations when choosing the approach. METHODS: A retrospective chart review of all consecutive patients undergoing inguinal hernia repair from January 2005 to December 2009 at a single institution was conducted. Presentation characteristics and outcome measures including recurrence rates, post-operative pain and complications were compared in women undergoing laparoscopic versus open hernia repair. RESULTS: A total of 1,133 patients had an inguinal herniorrhaphy. Of these, 101 patients were female (9 %), with a total of 111 hernias. A laparoscopic approach was chosen in 44 % of patients. The majority of women (56 %) presented with groin pain as the primary symptom. Neither the mode of presentation nor the presenting symptoms significantly influenced the surgical approach. There were no statistically significant differences in hernia recurrence, post-operative neuralgia, seroma/hematoma formation or urinary retention between the two approaches (p < 0.05). A greater proportion of patients with bilateral hernias had a laparoscopic approach rather than an open technique (12 vs. 2 %, p = 0.042). CONCLUSIONS: Laparoscopic herniorrhaphy is as safe and efficacious as open repair in women, and should be considered when the diagnosis is in question, for management of bilateral hernias or when concomitant abdominal pathology is being addressed.

Type II antithrombin deficiency caused by a large in-frame insertion: structural, functional and pathological relevance.

BACKGROUND: The metastable native conformation of serpins is required for their protease inhibition mechanism, but also renders them vulnerable to missense mutations that promote protein misfolding with pathological consequences. OBJECTIVE: To characterize the first antithrombin deficiency caused by a large in-frame insertion. PATIENTS/METHODS: Functional, biochemical and molecular analysis of the proband and relatives was performed. Recombinant antithrombin was expressed in HEK-EBNA cells. Plasma and recombinant antithrombins were purified and sequenced by Edman degradation. The stability was evaluated by calorimetry. Reactive centre loop (RCL) exposure was determined by thrombin cleavage. Mutant antithrombin was crystallized as a dimer with latent plasma antithrombin. RESULTS: The patient, with a spontaneous pulmonary embolism, belongs to a family with significant thrombotic history. We identified a complex heterozygous in-frame insertion of 24 bp in SERPINC1, affecting strand 3 of ß-sheet A, a region highly conserved in serpins. Surprisingly, the insertion resulted in a type II antithrombin deficiency with heparin binding defect. The mutant antithrombin, with a molecular weight of 59 kDa, had a proteolytic cleavage at W49 but maintained the N-terminal disulphide bonds, and was conformationally sensitive. The variant was non-inhibitory. Analysis of the crystal structure of the hyperstable recombinant protein showed that the inserted sequence annealed into ß-sheet A as the fourth strand, and maintained a native RCL. CONCLUSIONS: This is the first case of a large in frame-insertion that allows correct folding, glycosylation, and secretion of a serpin, resulting in a conformationally sensitive non-inhibitory variant, which acquires a hyperstable conformation with a native RCL.

Detection of a novel reassortant epizootic hemorrhagic disease virus (EHDV) in the USA containing RNA segments derived from both exotic (EHDV-6) and endemic (EHDV-2) serotypes.

Epizootic hemorrhagic disease virus (EHDV) is a Culicoides-transmitted orbivirus that infects domestic and wild ruminants and is provisionally thought to be distributed throughout Africa, North America, Australia, East Asia and the Middle East. Historically, of the seven proposed serotypes of EHDV, only EHDV-1 and EHDV-2 have been reported from North America. In 2006, EHDV isolates were recovered from moribund or dead white-tailed deer (Odocoileus virginianus) in Indiana and Illinois that could not be identified as either EHDV-1 or EHDV-2 by virus neutralization tests or by serotype-specific RT-PCR. Additional serological and genetic testing identified the isolates as EHDV-6, a serotype that, although originally described from Australia, has recently been recognized as an emerging pathogen of cattle in Morocco, Algeria and Turkey. In 2007 and 2008, EHDV-6 was isolated again from white-tailed deer, this time in Missouri, Kansas and Texas, suggesting that the virus is capable of overwintering and that it may become, or already is, endemic in a geographically widespread region of the USA. Genetic characterization of the virus indicates that it is a reassortant, such that the outer capsid proteins determining serotype specificity (VP2 and VP5) are derived from exotic EHDV-6, whilst the remaining structural and non-structural proteins are apparently obtained from indigenous EHDV-2 (Alberta).

Distribution of bluetongue in the United States of America, 1991-2002.

Bluetongue virus (BTV) distribution in the United States of America (USA) is limited by the range of the vector Culicoides spp. Regional differences exist with the north-eastern states being free of BTV, while the central and north-western states are seasonally free of virus. Activity of the virus can be observed throughout the year in the southern USA. Serological evidence defining the distribution of BTV in selected regions of the USA is gathered regularly through serological surveys conducted on samples from slaughter cattle. From 1991 to 2002, ten serological surveys were completed. Results from Alaska, Hawaii, Michigan, Minnesota, New York, Wisconsin and New England consistently demonstrated a seropositive rate of less than 2%, confirming BTV-free status. Antibody against BTV was sporadically detected in cattle originating from states contiguous to the BTV-free regions. Additional information on BTV distribution in the USA is obtained through identification of BTV or BTV RNA in diagnostic, surveillance and export specimens submitted to the National Veterinary Services Laboratories. Results confirm that BTV serotypes 2, 10, 11, 13 and 17 are present in the USA.

Characterizing the influence of structure and route of exposure on the disposition of dithiocarbamates using toluene-3,4-dithiol analysis of blood and urinary carbon disulfide metabolites.

Differences in the toxicities observed for dithiocarbamates have been proposed to result from the influence of nitrogen substitution, oxidation state, and route of exposure. To better characterize the fate of dithiocarbmates in vivoas a function of structure and route of exposure, rats were administered equimolar doses of carbon disulfide (CS2), N-methyldithiocarbamate, pyrrolidine dithiocarbamate, N,N-diethyldithiocarbamate, or disulfiram daily for five days, either po or ip, and sequential blood samples obtained. Protein dithiocarbamates formed by the in vivo release of CS2, parent dithiocarbamate, and protein-bound mixed disulfides were assessed in plasma and hemolysate by measuring toluene trithiocarbonate generated upon treatment with toluene-3, 4-dithiol (TdT). To aid in determining the bioavailability of CS2 from the administered dithiocarbamates, the urinary CS2 metabolites, 2-thiothiazolidine-4-carboxylic acid (TTCA) and 2-thiothiazolidin-4-ylcarbonylglycine (TTCG), were also determined. The levels of TdT-reactive moieties detected depended upon both the compound administered and the route of exposure. Parent dithiocarbamates, with the exception of disulfiram, were eliminated from blood within 24 h; but protein associated TdT-reactive moieties persisted and accumulated with repeated exposure, regardless of the route of exposure. N-Methyldithiocarbamate demonstrated the greatest potential to produce intracellular globin modifications, presumably through its unique ability to generate a methylisothiocyanate metabolite. Urinary excretion of TTCA and TTCG was more sensitive than TdT analysis for detecting dithiocarbamate exposure, but TdT analysis appeared to be a better indicator of in vivo release of CS2 by dithiocarbamates than were urinary CS2 metabolites. These data suggest that CS2 is a more important metabolite, following oral exposure, than are other routes of exposure, e.g., inhalation or dermal. In addition, data also suggest that acid stability, nitrogen substitution, and route of exposure are important factors governing the toxicity observed for a particular dithiocarbamate.

Factor Xa and thrombin, but not factor VIIa, elicit specific cellular responses in dermal fibroblasts.

UNLABELLED: Coagulation factors (F)VIIa, FXa and thrombin are implicated in cellular responses in vascular, mesenchymal and inflammatory cells. Fibroblasts are the most abundant cells in connective tissue, and damage to blood vessels places coagulation factors in contact with these and other cell types. OBJECTIVES: To investigate cellular responses of primary dermal fibroblasts to FVIIa, FXa and thrombin by following changes in expression of candidate proteins: monocyte chemotactic protein-1 (MCP-1), interleukin-8 (IL-8), interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF), and to determine the expression of receptors implicated in signaling by these coagulation factors. METHODS: Steady-state mRNA levels were quantified by RNase protection assay, and protein secretion by ELISA. PAR gene expression was assessed by ribonuclease protection assay and conventional and quantitative reverse-transcription-polymerase chain reaction. RESULTS: FVIIa did not induce the candidate genes. In contrast, FXa and thrombin induced MCP-1 mRNA and protein secretion strongly, IL-8 moderately, and IL-6 weakly. Neither FXa nor thrombin induced VEGF mRNA or protein secretion, although FXa induced VEGF protein secretion in lung fibroblasts. Comparison of the presence of candidate receptors in the two fibroblast subtypes demonstrated higher levels of PAR-1 and PAR-3 in lung fibroblasts relative to their dermal counterparts and the additional expression of PAR-2. CONCLUSIONS: FXa and thrombin induce expression of MCP-1, IL-8 and IL-6, and distribution and expression of PARs on dermal fibroblasts is reduced relative to their lung counterparts. Tissue origin may influence the cellular response of fibroblasts to coagulation proteases.

Equine West Nile encephalitis, United States.

After the 1999 outbreak of West Nile (WN) encephalitis in New York horses, a case definition was developed that specified the clinical signs, coupled with laboratory test results, required to classify cases of WN encephalitis in equines as either probable or confirmed. In 2000, 60 horses from seven states met the criteria for a confirmed case. The cumulative experience from clinical observations and diagnostic testing during the 1999 and 2000 outbreaks of WN encephalitis in horses will contribute to further refinement of diagnostic criteria.

Detection of North American West Nile virus in animal tissue by a reverse transcription-nested polymerase chain reaction assay.

A traditional single-stage reverse transcription-polymerase chain reaction (RT-PCR) procedure is effective in determining West Nile (WN) virus in avian tissue and infected cell cultures. However, the procedure lacks the sensitivity to detect WN virus in equine tissue. We describe an RT-nested PCR (RT-nPCR) procedure that identifies the North American strain of WN virus directly in equine and avian tissues.

Stable recombinant expression and characterization of the two haemophilic factor VIII variants C329S (CRM(-)) and G1948D (CRM(r)).

In haemophilia A, the functional defect at the molecular level of most factor VIII (FVIII) missense mutations remains unknown. Site-directed mutagenesis of B domain-deleted FVIII cDNA (FVIIISQ) was used to introduce two mutations associated with severe cross-reacting material (CRM)-negative (FVIII-C329S) or mild/moderate CRM-reduced (FVIII-G1948D) haemophilia A. Wild-type (FVIIISQ-WT) and variant FVIIISQ proteins were successfully expressed after stable transfection in Chinese hamster ovary (CHO) cells, and partially characterized at the intracellular, molecular and functional levels. Reverse transcription polymerase chain reaction analysis confirmed that both transcription and mRNA processing appeared normal in CHO cells transfected with both the wild-type and two variant constructs. In contrast to FVIIISQ-WT, immunofluorescence analysis of both CRM(-) and CRM(r) variants showed intracellular FVIII accumulation within the rough endoplasmic reticulum, suggesting secretion defects in transfected CHO cells. Immunoblot analysis of the FVIIISQ variant proteins that were secreted showed that they were expressed as mixed populations of uncleaved 170 kDa polypeptides, processed 90 kDa heavy chains and 80 kDa light chains, similar to FVIIISQ-WT. Phenotypic analysis of the B domain-deleted FVIIISQ variants expressed in CHO cells correlated well with the patients' reduced FVIII activity and, in addition, surface plasmon resonance studies demonstrated that both missense mutations were associated with increased rates of A2 domain dissociation following thrombin activation. We conclude that the mutations found are responsible for the haemophilia A phenotype, through intracellular retention and decreased stability of the active cofactor FVIIIa.

Can costs be measured and predicted by modeling within a cooperative clinical trials group? Economic methodologic pilot studies of the radiation therapy oncology group (RTOG) studies 90-03 and 91-04.

PURPOSE: To (1) measure radiation therapy costs for patients in randomized controlled clinical trials, (2) compare measured costs to modeling predictions, (3) examine cost distributions, and (4) assess feasibility of collecting economic data within a cooperative group. METHODS: The Radiation Therapy Oncology Group conducted economic pilot studies for two Phase III studies that compared fractionation patterns. Expected quantities of Current Procedural Terminology (CPT) codes and relative value units (RVU) were modeled. Institutions retrospectively provided procedure codes, quantities, and components, which were converted to RVUs used for Medicare payments. Cases were included if the radiation therapy quality control review judged them to have been treated per protocol or with minor variation. Cases were excluded if economic quality review found incomplete economic data. RESULTS: The median and mean RVUs were within the range predicted by the model for all arms of one study and above the predicted range for the other study. CONCLUSION: The model predicted resource use well for patients who completed treatment per protocol. Actual economic data can be collected for critical cost items. Some institutions experienced difficulty collecting retrospective data, and prospective collection of data is likely to allow wider participation in future Radiation Therapy Oncology Group economic studies.

Routine, single-item screening to identify abusive relationships in women.

BACKGROUND: Abusive relationships are associated with several demographic factors and many clinical problems in women. However, practices often do not screen for abuse. METHODS: This is a descriptive study of 1526 women aged 19 to 69 years who completed a health survey in 31 office practices. The 53-item survey included a question designed to screen for an abusive relationship. Our analysis compared self-reported measures of symptoms (N = 13) and functional limitations (n = 6) of women who had abusive relationships with those who did not. We also examined the utility of using a constellation of clinical problems to identify risk for abuse. RESULTS: Women in abusive relationships were more likely to be poor (37% vs 14%; P < .001) and young (87% were younger than 51 years versus 69% of those who were not in such relationships; P < .001). They had twice as many bothersome symptoms (3.1 vs 1.7; P < .001) and functional problems (1.6 vs 0.8; P < .001). Approximately 40% (36/89) of low-income women with emotional problems were at risk for abuse versus only 6% (64/1025) of women with adequate financial resources and no emotional problems. However, because so many women were at low risk, almost twice as many in this group (n = 64) reported abusive relationships than in the high-risk group (n = 36). CONCLUSIONS: Women in abusive relationships have many symptoms and functional limitations. However, symptoms and clinical problems provide insufficient clues for abuse. It is better just to ask. A single-item screening question appears adequate for this purpose.

Bcl-2 accelerates multistep prostate carcinogenesis in vivo.

The impact of bcl-2 proto-oncogene expression on the pathogenesis and progression of prostate cancer was examined in a transgenic mouse model. Probasin-bcl-2 transgenic mice were crossed with TRAMP (TRansgenic Adenocarcinoma Mouse Prostate) mice that express the SV40 early genes (T/t antigens) under probasin control. Prostate size, cell proliferation, apoptosis, and the incidence and latency of tumor formation were evaluated. The double transgenic, probasin-bcl-2 X TRAMP F1 (BxT) mice exhibited an increase in the wet weight of the prostate. This was associated with an increase in proliferation, attributable to T/t antigens, and a decrease in apoptosis attributable to bcl-2. The latency to tumor formation was also decreased in the BxT mice compared to the TRAMP mice. The incidence of metastases was identical in both the TRAMP and BxT mice. Lastly, the incidence of hormone-independent prostate cancer was reduced in the BxT mice compared to the TRAMP mice. Together, these results demonstrate that bcl-2 can facilitate multistep prostate carcinogenesis in vivo.

Validation of a reverse transcriptase multiplex PCR test for the serotype determination of U.S. isolates of bluetongue virus.

Bluetongue (BT) is an arthropod-borne viral disease affecting ruminants primarily in tropical and temperate regions of the world. Of the 24 serotypes of BT virus (BTV) identified worldwide, five have been found in the United States. Serotype identification of BTV isolates is important to the epidemiology of the virus, but current methods are cumbersome. A single-tube multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) assay, previously developed for the serotype determination of U.S. BTV isolates, was evaluated. The determination of serotype was based on the size of the resultant amplified product. The procedure was evaluated using all 24 serotypes of BTV and nine serotypes of epizootic hemorrhagic disease virus (EHDV), a closely related orbivirus. Only the five U.S. serotypes of BTV were detected by the mRT-PCR. The assay was further tested using 132 BTV isolates originating from 24 western and southern states of the United States, from several different host species, spanning a period of 24 years. The serotypes of the isolates were determined by both a virus neutralization (VN) procedure and the mRT-PCR. Comparison of the mRT-PCR to the standard VN showed that the mRT-PCR successfully identified the serotypes of 130 of the isolates and was shown to be more reliable and specific than the VN assay.