Modeling cold curing of Pierce's disease in Vitis vinifera 'Pinot Noir' and 'Cabernet Sauvignon' grapevines in California.

Pierce's disease (PD) of Vitis vinifera grapevines is caused by the bacterium Xylella fastidiosa, a pathogen with a wide plant host range. Exposure of X. fastidiosa-infected plant tissue to cold temperatures has been shown to be effective at eliminating the pathogen from some plant hosts such as grapevines. This "cold curing" phenomenon suggests itself as a potential method for disease management and perhaps control. We investigated cold therapy of PD-affected 'Pinot Noir' and 'Cabernet Sauvignon' grapevine. In the fall, inoculated plants and controls of each cultivar were transported to each of four field sites in California (Foresthill, McLaughlin, Hopland, and Davis) that differed in the magnitude of cold winter temperatures. A model for progression of the elimination of plant disease in relation to temperature was conceptualized to be a temperature-duration effect, where temperatures below a particular threshold kill X. fastidiosa with increasing efficacy as the temperature decreases to some value <6?C. The temperature effect was modeled as a likelihood of a particular temperature killing the pathogen and is termed the ?killing index?. We developed a mathematical model for cold curing of grapevines inoculated with X. fastidiosa and calibrated the model with cold-curing data collected in a field study. Parameter estimation resulted in lowest sum of squared differences across all 10 trials to be low temperature below which the organism is killed (T(0)) = 6°C, number of hours to achieve 100% cure (N(100)) = 195 h, number of hours to achieve 10% cure (N(10)) = 20 h, and killing index (K(x)) = 0.45 for Pinot Noir and T(0) = 6°C, N(100) = 302 h, N(10) = 170 h, and K(x) = 0.41 for Cabernet Sauvignon. With the parameter estimates optimized by model calibration, the simulation model was effective at predicting cold curing in four locations during the experiment, although there were some differences between Hopland for Pinot Noir and Davis for Cabernet Sauvignon. Using historical temperature data, the model accurately predicted the known severity of PD in other grape-growing regions of California, suggesting that it may have utility in assessing the relative risk of developing PD in proposed new vineyard sites.

Hyperbaric spinal ropivacaine for cesarean delivery: a comparison to hyperbaric bupivacaine.

UNLABELLED: We evaluated the clinical efficacy and safety of spinal anesthesia with 0.5% hyperbaric ropivacaine compared with 0.5% hyperbaric bupivacaine for elective cesarean delivery. Sixty healthy, full-term parturients were randomly assigned to receive either 12 mg of 0.5% hyperbaric bupivacaine or 18 mg of 0.5% hyperbaric ropivacaine intrathecally. There were no significant differences in demographic or surgical variables or neonatal outcomes between groups. Onset time of sensory block to T10 or to peak level was later in the Ropivacaine group (P < 0.05). The median (range) peak level of analgesia was T3 (T1-5) in the Bupivacaine group and T3 (T1-4) in the Ropivacaine group. Time for sensory block to recede to T10 did not differ between groups. Duration of sensory block was shorter in the Ropivacaine group (188.5 +/- 28.2 min vs 162.5 +/- 20.2 min; P < 0.05). Complete motor block of the lower extremities was obtained in all patients. Ropivacaine also produced a shorter duration of motor blockade than bupivacaine (113.7 +/- 18.6 min vs 158.7 +/- 31.2 min; P < 0.000). The intraoperative quality of anesthesia was excellent and similar in both groups. Side effects did not differ between groups. Eighteen milligrams of 0.5% hyperbaric ropivacaine provided effective spinal anesthesia with shorter duration of sensory and motor block, compared with 12 mg of 0.5% hyperbaric bupivacaine when administered for cesarean delivery IMPLICATIONS: Eighteen milligrams of 0.5% hyperbaric ropivacaine provided effective spinal anesthesia with shorter duration of sensory and motor block, compared with 12mg of 0.5% hyperbaric bupivacaine when administered for cesarean delivery.

The prevalence of viral hepatitis (HAV, HBV and HCV) in the Christchurch community.

AIM: To determine the prevalence of hepatitis A (HAV), hepatitis B (HBV) and hepatitis C (HCV) in adults randomly selected from the Christchurch community. METHODS: A list of names was randomly generated from the Christchurch electoral roll and subjects were sequentially contacted and invited to participate. A blood sample was taken and tested for hepatitis A (IgG anti-HAV antibody), hepatitis B (HBsAg and anti-HBc) and HCV (anti-HCV antibody) using Abbott Elisa kits. Subjects positive for HBsAg were also tested for HBeAg/HBV DNA. Those positive for anti-HBc were tested for anti-HBs. HCV antibody positive samples were tested for HCV RNA using PCR. RESULTS: 1064 subjects (30.3% of those invited) participated in the study. The prevalence of HAV antibodies was 27.9%, and increased with age. The overall prevalence of HBV markers was 42/1064 (4.2%), and of these 0.3% were HBsAg positive and 3.9% were considered immune. No gender or ethnic differences in these proportions were observed. The seroprevalence of HVC antibody was 3/1064 (0.3%), two of whom were also PCR positive for HCV RNA. CONCLUSION: In the Christchurch community there was a high prevalence of antibodies to HAV, which increased with age. The prevalence of HBsAg and antibody to HCV were both low at 0.3%.

Seroprevalence of Helicobacter pylori in the adult population of Christchurch: risk factors and relationship to dyspeptic symptoms and iron studies.

AIM: To determine the prevalence of Helicobacter pylori infection in subjects randomly selected from the Christchurch population and to determine the risk factors and symptoms related to the infection. METHODS: A list of names was randomly generated from the 1996 electoral roll and subjects were sequentially contacted and invited to participate. A questionnaire on dyspeptic symptoms was completed and the subject's serum was analysed for H. pylori antibodies using the Roche method. Equivocal samples were retested by the Meridian method. RESULTS: One thousand and sixty-four subjects participated in the study. In four subjects results for H. pylori were indeterminate and these subjects were excluded from analysis. Of the remaining 1060 subjects, 254 (24.0%) were seropositive for H. pylori. The seropositivity in males (n=444) was 25.9% and in females (n=616) 22.6%. On multivariate analysis age, ethnicity, low income and smoking > 20 cigarettes per day were all independent predictors of H. pylori seropositivity. H. pylori positive subjects had shorter stature compared to those who were seronegative. The symptom scores for dyspepsia were similar in both the seropositive and seronegative subjects. In males the serum iron levels were lower in seropositive subjects but there were no significant differences in serum ferritin in either males or females between seropositive and seronegative subjects. CONCLUSION: H. pylori is a common infection in the Christchurch community with the prevalence increasing significantly with age. H. pylori positive subjects had shorter stature and in males lower serum iron levels were observed. Infection was not associated with an increased risk of dyspeptic symptoms.

Dissolution of gallstones with simvastatin, an HMG CoA reductase inhibitor.

The aim of this study was to determine whether 12 months of therapy with Simvastatin, an HMG CoA reductase inhibitor, would dissolve gallstones. Twenty-seven subjects entered the study, all had a fasting oral cholecystogram, ultrasound examination, and fasting serum lipids prior to therapy. In addition, 22 subjects had their gallbladder ejection fraction, after CCK, determined by radionucleotide scanning. Eleven subjects had the cholesterol saturation index (CSI) of bile calculated before and at the end of 12 months of therapy. Of the 27 subjects, 26 completed 12 months of treatment with Simvastatin 20 mg daily. There was a significant fall in the total serum cholesterol (27%, P < 0.0001), LDL cholesterol (31%, P < 0.0001), triglyceride (34%, P < 0.0001) but no change in HDL after 12 months of therapy. Simvastatin treatment resulted in a 28% fall in the CSI of bile at the end of therapy (P < 0.01). The concentrations of individual bile acids did not change with therapy, and apart from a slight but significant increase in arachidonate, there were no other significant changes in the fatty acid composition of the biliary phospholipids. After 12 months of Simvastatin therapy there was a small decrease in the gallstone diameter but complete dissolution of gallstones was not achieved in any subjects. In conclusion 12 months of therapy with Simvastatin was effective in lowering the serum lipids and the CSI of bile but was not effective in dissolving gallstones.

The use of enzyme-linked immunosorbent assay for the quantitation of Calloselasma rhodostoma (Malayan pit viper) venom and venom antibodies.

The specificity and sensitivity of an indirect and two (an 'ordinary' and a 'rapid') double sandwich enzyme-linked immunosorbent assay (ELISA) procedures for the quantitation of Calloselasma rhodostoma (Malayan pit viper) venom were examined. The three assays were equally sensitive and the accuracy of the assays was not substantially affected by individual variation in the venom composition. The specificity of the assays was examined against 26 venoms from snakes of the families Viperidae and Elapidae. While the double sandwich ELISA procedures were sufficiently specific to be used in the clinical immunodiagnosis of C. rhodostoma bite in Malaysia, the indirect ELISA procedure exhibited extensive cross-reactivity with other Malaysian pit viper venoms. Attempts were made to improve the specificity of the indirect ELISA procedure for the quantitation of C. rhodostoma venom. A 'low ELISA cross-reactivity' venom fraction (termed VF52) was isolated from C. rhodostoma venom by repeated Sephadex G-100 gel filtration chromatography. The indirect ELISA procedure using antibodies to VF52 as immunoreagent showed an improvement in specificity. The use of the indirect ELISA procedure for the detection of C. rhodostoma antibodies was also examined and the results show that the assay was sufficiently specific to be used for retrospective diagnosis of C. rhodostoma bite in Malaysia, in particular when VF52 was used as the coating antigen.

A competitive ELISA for lipoprotein(a).

A competitive ELISA for lipoprotein(a) (Lp(a)) is described. The method uses a commercially available polyclonal anti-Lp(a) antibody and an IgG biotinstreptavidin-horseradish peroxidase detection system. The method is simple and robust with an assay sensitivity of 0.7 ng/well (1.4 micrograms/l). The antibody cross-reactivity was 0.14% against LDL and 0.70% against plasminogen. The coefficients of variation obtained with control sera of 266 and 552 mg/l were: 5.0% and 4.6% (n = 6), respectively for the intraassay; and 10.8% and 9.5% (n = 16), respectively for the interassay. The method showed an excellent correlation with a commercial immunoradiometric assay (IRMA), y (ELISA) = 0.94x (IRMA) - 8, (r = 0.98). A recovery study in which a 200 mg/L standard and four plasma samples were diluted with different proportions of a low plasma sample, gave linear relationships and also confirmed the specificity of the antibody.

An automated solid-phase 17 alpha-hydroxyprogesterone ELISA method using a microtitre plate.

We report an automated ELISA method for the measurement of 17 alpha-hydroxyprogesterone in plasma samples. A rabbit antiserum raised against 17 alpha-hydroxyprogesterone-3-(O-carboxymethyl)oxime-bovine serum albumin is used for the assay and a homologous competitor 17 alpha-hydroxyprogesterone-3-(O-carboxymethyl)oxime-bovine thyroglobulin is coated onto a microtitre plate. A goat anti-rabbit IgG-horse radish peroxidase is used as a probe for this solid-phase assay. The assay exhibits good sensitivity, precision and accuracy. The method is used routinely for the management of patients with congenital adrenal hyperplasia.

An enzyme-linked immunosorbent assay (ELISA) for plasma progesterone: immobilised antigen approach.

A single extraction ELISA for plasma progesterone is described using the fixed antigen approach. Progesterone is covalently coupled to bovine thyroglobulin and adsorbed onto a 96-well microtitre plate in guanidine hydrochloride. The assay, performed on an automatic ELISA processor, follows an established methodology used for other steroid hormones analysed in this laboratory with concomitant advantages in assay standardisation, cost structure and result throughput. A comparison with an established RIA shows the assay to be rapid, of similar specificity and accuracy with a sensitivity of less than 0.5 nmol/l and is suitable for use in a routine endocrine laboratory for determination of luteal function.

A monoclonal antibody to prednisone: use of enzyme-linked immunosorbent assay (ELISA) for screening and characterization of antigenic determinants.

ELISA technology using immobilized steroid conjugates has allowed the rapid screening and characterization of hybridoma supernatants. Although cortisol-3CMO-BSA was the immunogen, a monoclonal antibody with exceptionally high cross-reactivity (greater than 1000%) to prednisone was obtained. Characterization of the antigenic determinants shows a requirement for overall glucocorticoid 21-hydroxyl and 17 alpha-hydroxyl groups with additional high specificity for the 1,2-double bond and 11-position. There is potential use for this antibody in the assay of prednisone.

A competitive enzyme-linked immunosorbent assay for plasma 11-deoxycortisol (17,21-dihydroxy-4-pregnene-3,20-dione).

An enzyme-linked immunosorbent assay (ELISA) for 11-deoxycortisol is described for the first time. 11-Deoxycortisol-thyroglobulin conjugate is adsorbed onto the wells of a 96-well ELISA plate and competes with 11-deoxycortisol in the standards or plasma extract for antibody binding sites. After washing, immobilized primary antibody is probed with peroxidase-labeled goat anti-rabbit IgG. The ELISA plate is further washed and o-phenylenediamine added, color developed and the absorbance read at 492 nm. The ELISA shows good agreement with our existing 11-deoxycortisol radioimmunoassay (RIA) and has similar specificity and performance which allow its use in the routine steroid laboratory for assessing pituitary adrenal function by the metyrapone test.

125I radioimmunoassay for 17 alpha-hydroxyprogesterone in plasma, for diagnosing and managing congenital adrenal hyperplasia.

This rapid, inexpensive, and sensitive radioimmunoassay (RIA) for plasma 17 alpha-hydroxyprogesterone involves radioiodination. A single extraction with toluene/hexane removes an average 93% of the hormone from 0.1 mL of plasma. The extract is evaporated and the hormone is estimated by a simple, precise, and accurate 125I RIA involving a specific rabbit antiserum. A suspension of dextran-coated charcoal is used to separate free and bound steroid. Inter- and intra-assay CVs were less than 15 and less than 10%, respectively, and the sensitivity was 3 pg per assay tube. The regression equation for data on 17 alpha-hydroxyprogesterone added to steroid-free plasmas was y = 0.94x + 2.2 (r = 0.99). However, the turnaround time is only one-half to one-tenth that for most 3H RIA (3 h vs 6 to 30 h). The ranges of values found for plasma from normal subjects, treated and untreated patients with congenital adrenal hyperplasia, and infants with newly detected congenital adrenal hyperplasia were, respectively, 1 to 11, 0 to 20, 30 to 620, and 270 to 4900 nmol/L.