Evaluation of enrichment media for improved PCR-based detection of V. cholerae and V. vulnificus from estuarine water and plankton.

AIMS: Pathogenic Vibrio spp., including V. cholerae and V. vulnificus, are commonly found along the estuaries of the south-east United States; however, it is often difficult to recover these species directly from environmental samples. Pre-enrichment assays are commonly used to improve the detection of pathogenic vibrios from environmental sources. Here, we evaluated a novel enrichment procedure using freshly collected and autoclaved natural estuarine water amended with 1% peptone (designated as estuarine peptone water, EPW) and compared it to traditional alkaline peptone water (APW) for detection by PCR of V. cholerae and V. vulnificus. METHODS AND RESULTS: Of the 50 samples collected in total, V. cholerae DNA was detected in APW 10% of the time and in EPW 40% of the time. Likewise, the cholera toxin gene (ctxA) was detected in 4 vs 18% of the samples using APW and EPW, respectively. Conversely, APW showed improved recovery for V. vulnificus relative to EPW with respective detection frequencies of 46 and 20%. Results showed similar patterns across different sample types (water and plankton). CONCLUSIONS: While enrichment in traditional APW was adequate for the recovery of Vibrio vulnificius, use of sterile estuarine water amended with peptone significantly improved the detection of V. cholerae and the virulence gene ctxA from estuarine sources.

Persistence of anti-zonae pellucidae antibodies following a single inoculation of porcine zonae pellucidae in the domestic equine.

In this study of equids, we investigated the antibody response and the effect on the estrous cycle following a single inoculation of porcine zonae pellucidae (pZP) employing controlled-release methodology. We also investigated the use of two different water-soluble adjuvants as an alternative to oil-based adjuvants. Twenty-seven domestic mares were inoculated with various formulations of pZP and adjuvant. We showed that the anti-pZP antibodies generated as a result of the inoculations persisted for at least 43 weeks (length of the study). Of the various formulations used in the study, pZP and QS-21 water-soluble adjuvant, administered in combination with an emulsified preparation of pZP and Freund's Complete Adjuvant generated a significantly (P < 0.05) higher titer of anti-pZP antibodies when compared with other formulations employing the water-soluble adjuvant, Carbopol. Hormone analyses for cyclicity indicated a high incidence and extended duration of persistent corpora lutea among the treated mares. The positive control group of mares receiving two standard inoculations of pZP and Freund's Complete and Incomplete Adjuvants, as well as the placebo group of mares injected with QS-21 only, also exhibited high incidences of persistent corpora lutea. However, all mares eventually returned to normal cyclicity. The basis for the high incidence and extended duration of persistent corpora lutea was unexplained. The results demonstrate for the first time the persistent generation of anti-pZP antibodies following a single inoculation of pZP incorporated into a controlled-released preparation in the horse. This study further suggests that a single inoculation of pZP sequestered in a controlled-release lactide-glycolide polymer may serve as an alternative to traditional two-inoculation protocols for contraception investigations in the equine.

Field testing of immunocontraception on white-tailed deer (Odocoileus virginianus) on Fire Island National Seashore, New York, USA.

Application of contraception for the control of suburban populations of white-tailed deer (Odocoileus virginianus) has been much debated, but few data are available on field applications and even fewer on population effects. Between 1993 and 1997, 74-164 individually known female deer living on Fire Island, New York, USA, were treated remotely with an initial shot of 65 microg porcine zona pellucida (PZP) in Freund's complete adjuvant followed by booster injections of 65 microg PZP in Freund's incomplete adjuvant. Starting in 1996, progressively increasing numbers of deer were treated with vaccinating/marking darts. Estimates of population density and composition, using distance sampling methods, began in 1995 in selected portions of the study area. Between 1993 and 1997, fawning rates among individually known, treated adult females decreased by 78.9% from pretreatment rates. Population density in the most heavily treated area increased by 11% per year from 1995 to March 1998 and then decreased at 23% per year to October 2000. In 1999-2000 surveys, fawns comprised 13-14% of the total population in the most heavily treated area, versus 16-33% in nearby untreated areas. These results show that PZP can be delivered effectively to sufficient deer to affect population density and composition in some environments, but that technical and logistical improvements are needed before contraception can be used widely to manage suburban deer populations.

Porcine zona pellucida (PZP) immunocontraception of wild horses (Equus caballus) in Nevada: a 10 year study.

Porcine zona pellucida (PZP) immunocontraception was investigated for possible use in free-roaming wild horses in the western USA. A protocol of two injections (3-4 weeks apart) of vaccine lasting 1 year was first used and a single-injection controlled-release vaccine of 1 year duration was developed and tested in the field. Studies of a presumptive vaccine of 2 year duration were initiated. The parameters of anti-PZP antibody titre response, pregnancy testing and offspring production were used, and PZP vaccine was found to provide up to 94% infertility in free-roaming wild mares. In addition, a single-injection PZP vaccine of 1 year duration and containing a controlled-release component of PZP in a polymer matrix can provide infertility equivalent to the two-injection PZP vaccine. All the PZP vaccine preparations tested were associated with a return to normal fertility within 1 year. During the course of these studies, attention was given to practical aspects of management application of PZP contraception. Preparation of the controlled-release portion of the vaccine in pellets, which fit into the needle of a dart or syringe, has simplified vaccine handling and permitted long-term storage of the controlled-release component. Vaccine delivery is now performed using a jabstick on captured mares restrained in a field stock chute during routine horse gathers. Provision of a vaccine-training programme has maximized personnel safety during vaccine preparation and use.

Media richness and social information processing: rationale for multifocal continuing medical education activities.

Academic business communication has studied the results of media selection in organizations. Little of this work has been discussed in the context of continuing medical education (CME); however, it may apply to improving the design of educational activities. This article reviews literature on media richness and social information processing theories. The concept of media richness suggests that media choice results from a match between the objective characteristics of the medium and the content requirements of a message. In this context, media include face-to-face conversation and print and electronic media. Social information processing theory suggests that media selection is also based on participants' social norms for how information is communicated in their environment and the participants' familiarity with specific media types. Appraisal of CME with respect to these theories suggests that the complex relationship of CME content and CME participant environments invites the most effective strategies of multiple media experienced over time in what might be called multifocal continuing medical education.

Stable association of hsp90 and p23, but Not hsp70, with active human telomerase.

The ribonucleoprotein telomerase holoenzyme is minimally composed of a catalytic subunit, hTERT, and its associated template RNA component, hTR. We have previously found two additional components of the telomerase holoenzyme, the chaperones p23 and heat shock protein (hsp) 90, both of which are required for efficient telomerase assembly in vitro and in vivo. Both hsp90 and p23 bind specifically to hTERT and influence its proper assembly with the template RNA, hTR. We report here that the hsp70 chaperone also associates with hTERT in the absence of hTR and dissociates when telomerase is folded into its active state, similar to what occurs with other chaperone targets. Our data also indicate that hsp90 and p23 remain associated with functional telomerase complexes, which differs from other hsp90-folded enzymes that require only a transient hsp90.p23 binding. Our data suggest that components of the hsp90 chaperone complex, while required for telomerase assembly, remain associated with active enzyme, which may ultimately provide critical insight into the biochemical properties of telomerase assembly.

Patient satisfaction with telemedicine in a prison environment.

Prison inmates were surveyed about their perceptions of the use of videoconferencing in clinical consultations. A 14-item questionnaire was used to assess satisfaction with the patient-physician clinical interaction. Of the 299 inmates surveyed immediately after their teleconsultations, 221 completed questionnaires that were suitable for analysis (74%). Only 9% of patients indicated that they were not satisfied with the teleconsultation. They rated the telemedicine context well above the midpoint of the scale (16) on both an information-exchange and a patient-comfort dimension, with means of 29 and 24, respectively. There was a significant difference (P < 0.01) in the patient-comfort dimension in terms of the location of the prisoners. No significant differences were found in patient satisfaction related to the specialty of the physician. The results suggest that many contextual factors must be considered to understand the communicative implications of patient satisfaction with telemedicine.

Sire marbling score expected progeny difference and weaning weight maternal expected progeny difference associations with age at first calving and calving interval in Angus beef cattle.

Field records from the American Angus Association were used to study the associations of sire marbling score EPD and sire weaning weight maternal (milk) EPD with age at first calving (AFC) and calving interval (CI). Cows were selected based on the accuracy of their sire's milk (> or =.7) or marbling (> or =.6) EPD. The data were screened using biological constraints, and regression models were used to identify records that were greater than 5 SD from the mean. The AFC was modeled for both milk and marbling data sets to account for effects of year, sire EPD, and their interaction. The CI was subdivided into first, second, and mature calving interval traits and modeled to account for state, year, calf sex, calf birth weight (BW), calf weaning weight (WW), sire EPD, and interactions of EPD with year and state. Derivative-free REML was used to estimate heritability and genetic correlations for AFC and CI. Sire milk EPD and marbling EPD were predictors of AFC (P < .001); however, pooled estimates were unreliable because of state x EPD interactions (P < .001). Increases in sire milk EPD resulted in reductions in AFC; however, there was no consistent pattern to effects of marbling EPD increases. Models accounted for < 8% of variation in AFC. Sire milk EPD was not a predictor of first, second, or mature CI (P > .1). Sire marbling score EPD was not a predictor of second, or mature CI (P > .1); however, it was associated (P = .059) with first CI, although regression estimates varied across states and prevented pooling. The BW, sex, and WW were predictors of CI (P < .001). Increases in BW resulted in longer mature CI, and mature CI decreased as WW increased. The AFC was heritable (.22), and CI traits had heritabilities ranging from .01 to .03. The AFC was genetically correlated with first CI (-.6) and mature CI (-.93). Genetic correlations between CI traits were uninterpretable because of low additive genetic variances. In conclusion, sire marbling score and milk EPD do not seem to be reliable predictors of AFC or CI. The BW and WW have significant but small effects on AFC and CI. Selection for AFC is possible, but earlier calving heifers may have longer calving intervals.

Flexural properties and surface finishing of acetal resin denture clasps.

PURPOSE: This study examined flexural properties of acetal resin to determine the appropriate design for an acetal resin removable partial denture clasp. The effectiveness of various acetal resin polishing procedures was also evaluated using scanning electron microscopy. MATERIALS AND METHODS: Loads of up to 1,500 g were applied to cylindrical acetal resin specimen lengths of 5, 10, and 15 mm, and the degree of deflection of the specimen tip was measured. Specimens were 2 mm and 1.5 mm in diameter with a round cross-section (R2 and R1.5, respectively), as well as 2 mm in diameter and half-round in cross-section. Half-round specimens were loaded with their flat surface vertical and then horizontal (HRV and HRH, respectively). Contour plots showing load and deflection for varying lengths of resin were developed for each cross-sectional type based on specimen length, and the stiffness was calculated. Additional acetal resin specimens underwent polishing techniques including tungsten carbide burs, aluminium oxide stones, abrasive impregnated rubber points, and "Polish-D." Polished specimens were examined with scanning electron microscopy to compare surface roughness. RESULTS: For the 5-mm-length specimens, the median stiffness values decreased in the following order: R2 > HRV > HRH > R1.5. The median stiffness values for the 10-mm and 15-mm lengths followed a similar trend. At each length, the R2 specimen was generally at least twice as stiff as other designs. Regarding polishing, the smoothest surface was seen on the specimen that was polished with the rubber point followed by "Polish-D." CONCLUSIONS: To have stiffness similar to a cast cobalt-chromium clasp 15 mm long and 1 mm in diameter, a suitable acetal resin clasp must be shorter (approximately 5 mm) with greater cross-sectional diameter (approximately 1.4 mm). To obtain a smooth surface, the acetal resin clasp should be polished with a rubber point followed by a material such as "Polish-D."

Telemedicine: emerging e-medicine.

This paper reviews the emergence of telemedicine and its recent expansion and use within the healthcare industry. Through this review, several examples of telemedicine within a variety of applications provide a broad context to discuss the challenges and opportunities facing the emergence of e-medicine. These examples provide snapshots of a teleradiology system used by the military, teleconsultations used in neurosurgery and hemodialysis, and home telemedicine used in diabetes care. Based on the discussion of telemedicine's history and expansion and the examples provided, a framework is offered for understanding the evolution of telemedicine applications through four stages. These stages include: (a) development of basic technological capabilities, (b) development of relevant applications, (c) the integration of technical applications within a complex environment, and (d) transformation of the operating environment. Implications for this framework are discussed.

Candidate gene analysis of GH1 for effects on growth and carcass composition of cattle.

We present an approach to evaluate the support for candidate genes as quantitative trait loci (QTLs) within the context of genome-wide map-based cloning strategies. To establish candidacy, a bacterial artificial chromosome (BAC) clone containing a putative candidate gene is physically assigned to an anchored linkage map to localise the gene relative to an identified QTL effect. Microsatellite loci derived from BAC clones containing an established candidate gene are integrated into the linkage map facilitating the evaluation by interval analysis of the statistical support for QTL identity. Permutation analysis is employed to determine experiment-wise statistical support. The approach is illustrated for the growth hormone 1 (GH1) gene and growth and carcass phenotypes in cattle. Polymerase chain reaction (PCR) primers which amplify a 441 bp fragment of GH1 were used to systematically screen a bovine BAC library comprising 60,000 clones and with a 95% probability of containing a single copy sequence. The presence of GH1 in BAC-110R2C3 was confirmed by sequence analysis of the PCR product from this clone and by the physical assignment of BAC110R2C3 to bovine chromosome 19 (BTA19) band 22 by fluorescence in situ hybridisation (FISH). Microsatellite KHGH1 was isolated from BAC110R2C3 and scored in 529 reciprocal backcross and F2 fullsib progeny from 41 resource families derived from Angus (Bos taurus) and Brahman (Bos indicus). The microsatellite KHGH1 was incorporated into a framework genetic map of BTA19 comprising 12 microsatellite loci, the erythrocyte antigen T and a GH1-TaqI restriction fragment length polymorphism (RFLP). Interval analysis localised effects of taurus vs. indicus alleles on subcutaneous fat and the percentage of either extractable fat from the Iongissimus dorsi muscle to the region of BTA19 harbouring GH1.

Effectiveness of telemedicine in the outpatient pulmonary clinic.

BACKGROUND: Over the past several years, there has been a resurgence of interest in telemedicine. Despite this renewed interest, some health care providers remain skeptical regarding the effectiveness of telemedicine for the delivery of health care. OBJECTIVE: The objective of this prospective, crossover study was to determine if there was any difference between care delivered using video conferencing-based telemedicine technology and that given by a traditional face-to-face encounter in a pulmonary medicine clinic. METHODS: Two pulmonologists sequentially examined 40 individuals via video conferencing-based telemedicine technology and by the traditional face-to-face method. Two additional pulmonologists, in a blinded fashion, compared the results for consistency in the history, examination with focus on auscultation of the lungs, diagnostic impression, and evaluation and treatment plans. RESULTS: Evaluation of patients by telemedicine was as effective as the traditional mode. The telemedicine physician and the physician examining the patient in the traditional manner were able to elicit the same key complaints and hear the same adventitious sounds on auscultation of the lungs. CONCLUSION: Telemedicine can enable the provision of high-quality care in a pulmonary clinic setting.

The development and partial assessment of the medical communication competence scale.

The purpose of this research was to develop and partially assess a self-report scale for measuring doctors' and patients' perceptions of self-communication and other communication competence during a medical interview. Previous research into the components of communication competence and medical discourse were used to develop the Medical Communication Competence Scale (MCCS). It was hypothesized that the items of the MCCS would form four clusters: information giving, information seeking, information verifying, and socioemotional communication. The cluster analysis results provided support for the hypothesis. Results of several other analyses provided additional support for the validity of the MCCS.

A medium density microsatellite map of BTA10: reassignment of INRA69.

We have developed a genetic map of BTA10 based on 8952 informative meioses for 13 microsatellite markers and the erythrocyte antigen Z. With the exception of OarAE64, the support for the order of all loci in the map exceeded a LOD > 3.0. The length of the BTA10 genetic map was 87.0 centimorgans (cM). The 14-marker, sex-average map in Kosambi cM was: CSSM38-8.9-BM1237-5.2-HH8A-2.6- INRA69-10.6-TGLA378-0.8-BM6305-17.2- TGLA102-17.9-INRA96-0.3-CSRM60-9.2- DIK20-3.0-EAZ-6.7-CSSM46-3.7-SRCRSP3-1.0-OarA E64 with an average interval of 6.70 cM. The microsatellite INRA69 was recently assigned to the pseudoautosomal region of the bovine X chromosome by linkage analysis. However, we found that twopoint support for linkage between INRA69 and 15 X-linked bovine microsatellites was LOD < 0.50 in 529 reciprocal backcross and F2 fullsib progeny. We performed twopoint analyses of INRA69 against 275 markers distributed throughout the bovine genome and found significant associations with a LOD > 3.0 only between INRA69 and eight BTA10 microsatellite loci. Consequently, we excluded INRA69 from the genetic map of the X chromosome and reassign this microsatellite to BTA10.

Case studies in wildlife immunocontraception: wild and feral equids and white-tailed deer.

Non-lethal management methods are required for wild equids that are protected by law and for deer inhabiting areas where lethal controls are not legal or safe. Single or multiple inoculations of porcine zona pellucida (PZP) vaccine have been delivered to wild horses and deer by means of darts. Contraceptive efficacy in horses after two inoculations ranged from 90% to 100%, and after a single inoculation ranged from 19% to 28%. Mares given a controlled-release form of the vaccine had foaling rates ranging from 7% to 20%. No detectable changes in social organization or behaviours among treated horses occurred. Contraceptive effects were reversible after 4 consecutive years of treatment but 5-7 years of treatment resulted in ovulation failure and decreased urinary oestrogen concentrations. Among deer, two inoculations were 70-100% effective in preventing fawns, but one inoculation yielded a contraceptive efficacy of < or = 20%, with pregnancies occurring late in the breeding season; a single annual booster inoculation reduced fertility to 20% in the second year. Energy costs of extended breeding seasons were less than those resulting from pregnancy. After two years of treatment, ovaries appeared normal. These studies suggest that PZP immunocontraception can be successfully applied to certain free-roaming populations of wild horses and deer.

Remotely delivered immunocontraception in free-roaming feral burros (Equus asinus).

Regulation of local overpopulations of free-roaming feral equids is in demand worldwide for ecological balance and habitat preservation. Contraceptive vaccines have proven effective in feral horses, which breed seasonally, but no data are available for equids such as the burro, which is reproductively active all year round. In the present study, 27 individually identified female feral burros (Equus asinus) roaming free in Virgin Islands National Park (St John, US Virgin Islands; Lesser Antilles) were remotely treated with pig zonae pellucidae (PZP) vaccine. Between January and May, 16 burros were darted with a 1 ml emulsion of PZP plus Freund's adjuvant. Ten to twelve months later each treated burro was given a single booster injection of PZP plus adjuvant to maintain contraception through a second year. Eleven adult untreated jennies served as controls. Beginning one year after initial vaccination, these burros were monitored for pregnancy and foal production. Collection of data to determine treatment effect was not begun until 12 months after initial treatment to ensure that pregnancies existing before vaccination were not included. Pregnancy was assessed using previously validated methods for steroid metabolite measurement in fresh faecal samples. None of the PZP-treated burros produced foals between 0 and 12 months after the last inoculation. One PZP-treated burro tested positive for pregnancy at 10 months after the final inoculation. During this same period, six of 11 untreated burros tested pregnancy-positive, and four were observed with foals. There was no difference in pregnancy rates among treated, control and randomly sampled jennies between 12 and 24 months after the last inoculation. The results demonstrate that, in free-roaming feral burros that are reproductively active all year round: (1) burros can be accessed for remotely delivered PZP vaccination; (2) PZP contraception is effective; (3) PZP contraception is reversible; and (4) pregnancy can be reliably detected by faecal steroid analysis.

Palladium-103: a new radioactive source in the treatment of unresectable carcinoma of the pancreas: a phase I-II study.

Palladium-103 (Pd-103) is introduced in brachytherapy procedures because of its favorable physical properties, including its low energy, rapid dose fall-off, short half-life, and total cumulative dose delivery at a higher dose rate than iodine-125 (I-125) isotope. Intraoperative brachytherapy using I-125 pellets was reported to provide significant palliation and meaningful prolongation of life in highly selected patients with unresectable carcinoma of the pancreas. After considering some of the advantages of Pd-103 over I-125, we designed a phase I-II clinical trial to assess the feasibility of intraoperative Pd-103 in unresectable carcinoma of the pancreas to study the related morbidity when combined with chemotherapy and external beam radiation, and to evaluate the impact on palliation and local control rates. Between December 1989 and December 1993, 15 patients with biopsy-proven unresectable adenocarcinoma of the pancreas were treated with interstitial Pd-103 implants during laparotomy. In 13 patients the lesion was located in the head of the pancreas, in one patient in the uncinate process, and in one patient in the body of the pancreas. The stage distribution was as follows: T1 = 2; T2 = 6, and T3 = 7. In addition, all patients underwent biliary and gastric bypass. The mean number of Pd-103 pellets was 45; the mean total activity to obtain a matched peripheral dose (MPD) of 11,000 cGy was 68.9 mCi. The mean tumor volume encompassing the MPD was 16.5 cc. All patients received postoperative external beam radiation (4,500 cGy over 4 1/2 weeks) and chemotherapy (5-fluorouracil and mitomycin C). This combined treatment, consisting of intraoperative brachytherapy using Pd-103 and postoperative external beam radiation with chemotherapy, was well tolerated in all patients. These were no treatment-related mortalities, and no serious complications, such as bleeding or fistula formation. Pain relief was obtained within 3-6 weeks in 10 out of 12 patients presenting with pain. Survival ranged from 6 to 24 months (median 10 months). The study suggests that Pd-103 can be considered an alternative to I-125 for interstitial brachytherapy for unresectable carcinoma of the pancreas. Symptom relief appeared to occur faster and complications are significantly less. However, this study did not show any improvement in the median survival rate over I-125 due to the advanced stage cancer in the majority of patients in the study.