Environmental investigations of Vibrio parahaemolyticus in oysters after outbreaks in Washington, Texas, and New York (1997 and 1998).

Total Vibrio parahaemolyticus densities and the occurrence of pathogenic strains in shellfish were determined following outbreaks in Washington, Texas, and New York. Recently developed nonradioactive DNA probes were utilized for the first time for direct enumeration of V. parahaemolyticus in environmental shellfish samples. V. parahaemolyticus was prevalent in oysters from Puget Sound, Wash.; Galveston Bay, Tex.; and Long Island Sound, N.Y., in the weeks following shellfish-associated outbreaks linked to these areas. However, only two samples (one each from Washington and Texas) were found to harbor total V. parahaemolyticus densities exceeding the level of concern of 10,000 g(-1). Pathogenic strains, defined as those hybridizing with tdh and/or trh probes, were detected in a few samples, mostly Puget Sound oysters, and at low densities (usually <10 g(-1)). Intensive sampling in Galveston Bay demonstrated relatively constant water temperature (27.8 to 31.7 degrees C) and V. parahaemolyticus levels (100 to 1,000 g(-1)) during the summer. Salinity varied from 14.9 to 29.3 ppt. A slight but significant (P < 0.05) negative correlation (-0.25) was observed between V. parahaemolyticus density and salinity. Based on our data, findings of more than 10,000 g(-1) total V. parahaemolyticus or >10 g(-1) tdh- and/or trh-positive V. parahaemolyticus in environmental oysters should be considered extraordinary.

Evaluation of nonisotopic DNA hybridization methods for detection of the tdh gene of vibrio parahaemolyticus.

Production of the thermostable direct hemolysin (TDH) by Vibrio parahaemolyticus is associated with pathogenicity of the organism and is encoded by the tdh gene. The timely resolution of seafood-associated outbreaks requires rapid and accurate detection of pathogenic V. parahaemolyticus. The specificity of alkaline phosphatase- and digoxigenin-labeled tdh gene probes was evaluated against 61 strains of V. parahaemolyticus (including isolates from recent outbreaks involving oysters from the Pacific Northwest, Texas, and New York), 85 strains of other vibrios, and 7 strains of non-vibrio species from clinical and environmental sources. The probes were specific for detection of the V. parahaemolyticus tdh gene.

Evaluation of alkaline phosphatase- and digoxigenin-labelled probes for detection of the thermolabile hemolysin (tlh) gene of Vibrio parahaemolyticus.

The biochemical identification and enumeration of Vibrio parahaemolyticus as described in the FDA Bacteriological Analytical Manual is expensive and labour-intensive. To reduce the time and effort necessary to verify the identity of V. parahaemolyticus, the use of a thermolabile haemolysin (tlh) gene probe is proposed. An alkaline phosphatase (AP)-labelled probe was evaluated for specificity against 26 strains of V. parahaemolyticus, 88 strains of other Vibrio species and 10 strains of non-vibrio species. Of the 124 isolates tested, the probe hybridized only with the 26 strains of V. parahaemolyticus, indicating species specificity. Two hundred and six suspect V. parahaemolyticus isolates from oysters were tested by this probe and API-20E diagnostic strips; there was 97% agreement between results. A digoxigenin (DIG)-labelled probe for detection of the tlh gene fragment was prepared by PCR and compared with the AP-labelled probe. When tested on 584 suspect V. parahaemolyticus isolates, results obtained with the AP- and DIG-labelled probes were in 98% agreement. These results suggest that the probes are equivalent for detection of the V. parahaemolyticus tlh gene.

Influence of water temperature and salinity on Vibrio vulnificus in Northern Gulf and Atlantic Coast oysters (Crassostrea virginica).

This study investigated the temperature and salinity parameters associated with waters and oysters linked to food-borne Vibrio vulnificus infections. V. vulnificus was enumerated in oysters collected at three northern Gulf Coast sites and two Atlantic Coast sites from July 1994 through September 1995. Two of these sites, Black Bay, La., and Apalachicola Bay, Fla., are the source of the majority of the oysters implicated in V. vulnificus cases. Oysters in all Gulf Coast sites exhibited a similar seasonal distribution of V. vulnificus: a consistently large number (median concentration, 2,300 organisms [most probable number] per g of oyster meat) from May through October followed by a gradual reduction during November and December to < or = 10 per g, where it remained from January through mid-March, and a sharp increase in late March and April to summer levels. V. vulnificus was undetectable (< 3 per g) in oysters from the North and South Carolina sites for most of the year. An exception occurred when a late-summer flood caused a drop in salinity in the North Carolina estuary, apparently causing V. vulnificus numbers to increase briefly to Gulf Coast levels. At Gulf Coast sites, V. vulnificus numbers increased with water temperatures up to 26 degrees C and were constant at higher temperatures. High V. vulnificus levels (> 10(3) per g) were typically found in oysters from intermediate salinities (5 to 25 ppt). Smaller V. vulnificus numbers (< 10(2) per g) were found at salinities above 28 ppt, typical of Atlantic Coast sites. On 11 occasions oysters were sampled at times and locations near the source of oysters implicated in 13 V. vulnificus cases; the V. vulnificus levels and environmental parameters associated with these samples were consistent with those of other study samples collected from the Gulf Coast from April through November. These findings suggest that the hazard of V. vulnificus infection is not limited to brief periods of unusual abundance of V. vulnificus in Gulf Coast oysters or to environmental conditions that are unusual to Gulf Coast estuaries.

Adjuvant radiation, chemotherapy, and androgen deprivation therapy for pathologic stage D1 adenocarcinoma of the prostate.

OBJECTIVES: A retrospective analysis of the results of an aggressive multimodal approach combining radical prostatectomy with adjuvant radiation, chemotherapy, or androgen deprivation therapy for patients with pathologic Stage D1 prostate carcinoma was performed to assess the impact of these therapies on survival, recurrence, local control, and morbidity. METHODS: Case records of 76 patients with pathologic Stage D1 tumors were reviewed. All had radical retropubic prostatectomy and were recommended adjuvant therapy based on the pathologic extent of the primary tumor and the number of involved lymph nodes. RESULTS: With a median follow-up of 7 years, overall survival was estimated to be 88% and 66% at 5 and 10 years, respectively, and equaled age- and race-matched controls. Prostate cancer-specific survival at 5 and 10 years was 88% and 74%, respectively. The probability of developing a clinically detectable recurrence (excluding prostate-specific antigen [PSA]) was 29% and 62% at 5 and 10 years, respectively. When PSA was added to the detection data, the probability of developing a recurrence increased to 58% and 78% at 5 and 10 years, respectively. Recurrence and cause-specific survival correlated with Gleason sum. Univariate analysis of the adjuvant therapies demonstrated no effect on survival, but adjuvant radiation alone and in combination with androgen deprivation increased the time to recurrence. Local control was excellent, surgical morbidity was equivalent to that of all patients undergoing prostatectomy during the same time period, and the morbidity of adjuvant therapy was minimal. CONCLUSIONS: Survival equivalent to age- and race-matched controls, with excellent control of the extensive primary tumor, can be achieved in patients with Stage D1 prostate carcinoma by a combination of radical prostatectomy and radiation therapy without the need for routine androgen deprivation therapy.

Effect of time and temperature on multiplication of Vibrio vulnificus in postharvest Gulf Coast shellstock oysters.

After harvest, shellstock oysters stored under controlled temperatures of 10, 13, and 18 degrees C and at ambient outside air temperature (23 to 34 degrees C) were sampled after 12 and 30 h for Vibrio vulnificus. At 13 degrees C and below, V. vulnificus failed to multiply in the oysters. In oysters held at 18 degrees C for 30 h and under ambient conditions for 12 and 30 h, V. vulnificus numbers were statistically greater (P < 0.05) than those in oysters at harvest. These data indicate that endogenous V. vulnificus can multiply in unchilled shellstock oysters.

Radical retropubic prostatectomy and postoperative adjuvant radiation for pathological stage C (PcN0) prostate cancer from 1976 to 1989: intermediate findings.

Between 1976 and 1989, 114 patients undergoing radical retropubic prostatectomy and bilateral pelvic lymph node dissection for prostatic adenocarcinoma were found to have stage PcN0 lesions. Postoperative adjuvant radiation therapy without hormonal treatment was given to 95 of these patients (83%): 26 (27%) with stage C1, 37 (39%) with stage C2 and 32 (34%) with stage C3 disease. The median radiation dose was 45 Gy. given at 180 cGy. daily. Median followup was 4.4 years (range 1.4 to 13.3). The overall 5 and 10-year actuarial rates for the patients were 94% and 70%, respectively. Disease-specific 5 and 10-year actuarial survival rates were 99% and 78%, respectively. At 5 and 10 years the chance of clinical recurrence was estimated as 6% and 13%, respectively, and the chance of recurrence (clinical or indicated by prostate specific antigen levels) was estimated to be 34% and 46%, respectively. Patients with high Gleason scores (8 to 10) and seminal vesicle involvement (stage C3) fared worst. There were 5 patients with clinical distant metastases, 1 with a clinical local recurrence and 1 with both conditions. Detectable elevation of prostate specific antigen without clinically evident recurrence was noted in 25 patients. Radiation therapy was well tolerated with minimal morbidity. Disease-specific survival and survival without clinical recurrence were improved over historical control in patients with stage PcN0 prostate cancer treated by radical prostatectomy alone. These data support a role for adjuvant radiation therapy in stage PcN0 prostate adenocarcinoma following radical prostatectomy.

Laryngeal flap for hypopharynx reconstruction.

Hypopharynx repair following resection of a pyriform sinus carcinoma may be accomplished primarily or involve a more extensive procedure, such as a free intestinal graft. Described herein is a laryngeal flap which may be used to repair a hypopharyngeal defect when a strip of mucosa, too narrow for a primary repair, is left. This method has been used successfully in 4 patients and has saved these patients from more extensive repairs.

Enzyme immunoassay for identification of Vibrio vulnificus in seawater, sediment, and oysters.

Historically, methods used to identify Vibrio vulnificus in environmental samples have been inadequate because isolation and identification procedures are time-consuming and fail to separate V. vulnificus from other bacterial species. We describe an enzyme immunoassay (EIA) and culture techniques which identified V. vulnificus in seawater, sediment, and oysters. The EIA used monoclonal antibody FRBT37 to a species-specific epitope of V. vulnificus. No cross-reactions were observed among 72 non-V. vulnificus strains comprising 34 species and 15 genera. In field trials, the EIA identified correctly 99.7% of 348 biochemically confirmed V. vulnificus isolates. The epitope corresponding to FRBT37 was found in cells lysed by Triton X-100, deionized H2O, and ultrasonication but was not found in culture supernatants, indicating that its location was intracellular. In addition, electron micrographs of V. vulnificus labeled with FRBT37-biotin-avidin-gold showed that epitope FRBT37 reacted with fragments of lysed cells but not whole cells. FRBT37 was expressed when V. vulnificus was cultured in different growth media. The minimum level of detection of the EIA was approximately 2,000 V. vulnificus cells per EIA well. Epitope FRBT37 was labile at 70 degrees C for 30 min. Immunoblot and EIA plate formats reduced assay time and facilitated handling large numbers of test samples.

Squamous cell carcinoma radioimmunoassay in squamous cell carcinoma of the head and neck.

Tumor-associated antigen has shown promise as a clinical aid in the detection and monitoring of uterine cervical squamous cell carcinoma. Antigen levels have been shown to reflect the extent of disease and response to treatment. These findings have suggested that measurements of tumor-associated antigen may be useful in monitoring other squamous cell carcinomas. To test this hypothesis, we measured tumor-associated antigen using the squamous cell carcinoma radioimmunoassay in 103 patients with previously treated squamous cell head and neck tumors and 28 patients with known squamous cell carcinoma of the head and neck. Increased squamous cell carcinoma antigen levels were found in 39 percent of patients with known tumors and in 19 percent of the patients with previous curative resection. The sensitivity of the assay limited its usefulness in predicting the presence of new and recurrent tumors.

Current status of pancreas transplantation.

Pancreas transplantation for the treatment of diabetes mellitus is being done with increasing frequency. Refined operative techniques, an improved immunosuppression regimen, and an earlier recognition of rejection have led to dramatic increases in both graft and patient survival rates. Preliminary data suggest that a functioning pancreatic allograft may arrest or reverse most of the complications of diabetes, although the effects on retinopathy remain controversial. Patients also acquire a strong sense of well-being after successful pancreas transplantation.

Hemilaryngeal reconstruction using an axial island cheek flap supported by Marlex and stainless steel wire mesh.

The defect that remains after an extended hemilaryngectomy continues to be a challenge to the reconstructing surgeon. The reconstruction ideally must provide airway protection against aspiration, allow for phonation, and provide a durable mucosal surface. It also must be accomplished in one stage. Nine Labrador dogs underwent successful reconstruction of the hemilarynx using an axial island cheek flap based on the facial artery and vein. Adequate laryngeal function was demonstrated by maintenance of body weight, normal barium swallows, return of strong bark, and no evidence of aspiration pneumonia. Pathologic review confirmed a viable mucosal surface and incorporation of the Marlex and stainless steel wire mesh in a fibrous reaction. We have concluded that this method of reconstruction provides a result superior to currently used techniques.

Mandibular reconstruction in an irradiated field using median sternotomy and anterior rib axial graft.

Mandibular reconstruction in an irradiated field continues to be a challenge to the reconstructing surgeon. Several methods use rib grafts to bring vascularized tissue to the mandibular region. A median sternotomy provides excellent exposure to harvest the internal mammary artery and anterior axial rib graft for subsequent mandibular reconstruction. The modified technique described provides an alternative method of bringing a vascularized rib graft to an irradiated field for mandibular reconstruction avoiding multiple rib resections and microvascular anastomosis. This technique has been used in two patients who had anterior mandibular resection and irradiation therapy for advanced squamous cell carcinoma. The results from this technique were excellent and without major complications.

Recommended modification of dilution procedure used for bacteriological examination of shellfish.

The dilution procedure recommended by the American Public Health Association for use in bacteriological analysis of shellfish samples may cause an underestimation of the number of bacteria in shellfish. Modifications of that procedure to include preparation of the first dilution of the shellfish homogenate on a weight rather than a volume basis, and the inoculation of 1 g portions of the shellfish from a dilution of the homogenate, have been shown to significantly increase the recovery of fecal coliform bacteria from oyster samples. It is recommended that these modifications be adopted by the American Public Health Association.

The relationship of rehabilitation center staff and client endorsement of appropriate client behaviors to outcome.

Clients who enroll in comprehensive rehabilitation centers but fail to complete their planned programs tend to be, at follow-up, less well adjusted than clients who do complete their planned program. Consequently, much research has been directed at understanding why clients fail and at identifying those clients most likely to fail. This study analyzed the relationship of client and staff expectations toward important client behaviors to outcome. Using a two-dimensional scaling technique, staff (n = 74) and clients (n = 268), at a large comprehensive rehabilitation center were asked to judge their acceptance of clients exhibiting either independent or dependent behaviors. Results indicated that both clients and staff approved of independence over dependence, that there was no difference between types of staff in their perceptions of appropriate client behaviors, that there were no significant differences between client program completers and noncompleters in perceptions toward appropriate behaviors, but that there were significant differences between clients and staff in the degree of approval of select client behaviors. Implications for rehabilitation practice and future research are discussed.

Automatic incubator for use with modified A-1 test for enumerating fecal coliform bacteria in shellfish growing waters.

An automatic water bath incubator which can be constructed from easily obtainable materials is described. This incubator provides temperatures necessary for both the 35 +/- 0.5 degrees C resuscitation period and the 44.5 +/- 0.2 degrees C final incubation period as required in the AOAC method for enumerating fecal coliforms by the modified A-1 test. Statistical analysis of experimental data indicated no significant differences (P greater than 0.50) in fecal coliform counts obtained by using the automatic incubator and the AOAC incubation method.

A multivariate analysis of motivational attributes among spinal cord injured rehabilitation clients.

Based on the assumption that understanding client motivation is central to the rehabilitation process but that motivation is complex, configural, and not a bipolar construct, this study sought to describe the motivational patterns of 110 spinal cord injured persons receiving vocational rehabilitation services. Self-report inventories purporting to measure four motivational concepts--arousal level, expectations, incentive conditions and need fulfillment--were used as descriptors. A cluster analysis procedure was used to form homogeneous subgroups of spinal cord injured persons according to motivational variable scores. Results indicated that the cluster analysis technique was reliable and that 80% of the sample was correctly classified into one or another of four subgroups on the basis of five motivational variables: personal goal choice and perceived importance of that goal, current life satisfaction, family need satisfaction, age, and arousal level. The clearest discriminator in forming the subgroups was type of goal chosen and perceived importance of that goal. Other demographic and psychological information was provided to sharpen intragroup similarities and intergroup differences. Implications for rehabilitation practice and future research were discussed.