A chemogenomic approach to understand the antifungal action of Lichen-derived vulpinic acid.

AIM: To determine uncovered antifungal activity of lichen-derived compound, vulpinic acid, by using chemical-genetic analyses. METHODS AND RESULTS: Haploinsufficiency and homozygous-profiling assays were performed, revealing that strains lacking GLC7, MET4, RFC2, YAE1 and PRP18 were sensitive to three concentrations (12·5, 25 and 50% of inhibitory concentration) of vulpinic acid and independently validated. To verify inhibition of those genes, cell cycle analysis using flow cytometry was performed and relative expressions were measured. Under vulpinic acid-treated condition, cell cycle was arrested in S and G2/M phases and sensitive strains' relative expressions were significantly lower than the wild type yeast. CONCLUSIONS: Vulpinic acid mainly affects cell cycle, glycogen metabolism, transcription and translation to fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Although lichen-derived compounds are commercially valuable, few studies have determined their modes of action. This study used a chemogenomic approach to gain insight into the mechanisms of one of well-known lichen-derived compound, vulpinic acid.

Antifungal effect of gamma irradiation and sodium dichloroisocyanurate against Penicillium expansum on pears.

UNLABELLED: Gamma irradiation (GI) was evaluated for its in vitro and in vivo antifungal activity against Penicillium expansum on pear fruits. GI showed a complete inhibition of spore germination, germ tube elongation and mycelial of P. expansum, especially 1·8 kGy. GI affected the membrane integrity and cellular leakage of conidia in a dose-dependent manner. Furthermore, the leakage of protein and sugar from mycelia increased along with the dose. GI was evaluated at lower doses in combination with a chlorine donor, sodium dichloro-s-triazinetrione (NaDCC), to examine the inhibition of P. expansum. Interestingly, only a combined treatment with 0·2 kGy of GI and 70 ppm of NaDCC exhibited significant synergistic antifungal activity. The mechanisms by which the combined treatment decreased the blue mould decay of pear fruits could directly associated with the disruption of the cell membrane of the fungal pathogen, resulting in a loss of cytoplasmic materials from the hyphae. SIGNIFICANCE AND IMPACT OF THE STUDY: Gamma irradiation (GI) is used as an effective nonchemical approach to inactive pathogens. This study investigated the antifungal effect of gamma irradiation and its combined treatment with a chlorine donor on this fungal pathogen, both in vitro and in vivo. This study emphasized that the integration of low-dose GI and a chlorine donor, NaDCC, exhibited a significant antifungal effect, and that its mechanisms are directly associated with membrane integrity of fungal spores, promising that GI has the potential to be an antifungal approach.

First Report of Leaf Spot in Farfugium japonicum Caused by Alternaria tenuissima in Korea.

Farfugium japonicum (L.) Kitam (common name: Leopard plant) is known as a medical herb and belongs to family Asteraceae (1). In June 2012, a leaf spot disease was observed on the leaf surface of F. japonicum at a forest research plot Jinju, Gyeongnam province, Korea. More than 95% of F. japonicum plants were infected and leaf spot symptom appeared in the regions under our investigation. Light brownish symptoms initially developed and the spot size gradually increased and turned dark brown with an irregular shape as the disease progressed (spot size 1 to 10 mm in diameter). At the late stage of disease, spots became hollow and completely dehydrated. The infected leaves were easily crumbled, possibly due to dryness. To isolate the causal agent, the infected leaves were surface disinfected and pieces of leaves were placed on water agar (WA). Nine isolates were isolated from 10 pieces of the infected leaves. Fungi mycelia from the WA were transferred on potato dextrose agar (PDA) and incubated at 28°C for 7 days. The colonies were purple navy to black and conidia spores developed on the media. The morphological characteristics of spores were multi-septate, dark brown, pyriform, and 6.7 to 12.8 × 22.2 to 38.4 µm. The spores had 1 to 4 transverse and 0 to 3 longitudinal septa. The morphological characteristics of the isolates showed considerably similar to well-known Alternaria tenuissima (2). The leaf spot disease caused by A. cinerariae of F. japonicum was reported from Japan (3). Spores of A. cinerariae are golden brown to brown with 3 to 9 transverse and 0 to 6 longitudinal septa and are 87.5 × 28.7 µm (avg.) (3). To verify pathogenicity of the isolate, the pure cultured fungi on the PDA medium was taken (4 mm in diameter) and placed on healthy leaves of Leopard plant. The artificially inoculated leaves were placed on wet filter paper in Petri dishes and incubated at 25°C and 80% humidity. At 7 days after inoculation, similar disease symptoms developed on 8 out of 10 infected Leopard plant leaves. The pathogen was reisolated from artificially infected leaves. To identify in molecular biology level, genomic DNA was extracted and the ITS-rDNA region was amplified using universal primers ITS1 and ITS4. The amplified PCR product was purified and sequenced (528 bp) with ITS1 and ITS4 primers for both directions and then deposited in GenBank (Accession No. KC415611.1). The BLAST search showed that it matched previously reported A. tenuissima with 100% identity. To the best of our knowledge, this is the first report of Leopard plant leaf spot disease in Korea. References: (1) E. Y. Kim et al. J. Ethnopharmacol. 146:40, 2013. (2) E. G. Simmons. Page 1 in: Alternaria Biology, Plant Diseases and Metabolites. J. Chelchowski and A. Visconti, eds. Elsevier, Amsterdam, 1992. (3) T. Sakoda et al. Res. Bull. Pl. Prot. Japan 46:73, 2010.

An Outbreak of Leaf Spot Caused by Corynespora cassiicola on Korean Raspberry in Korea.

In September and October 2010, leaf spots were observed on Korean raspberry (Rubus crataegifolius Bunge) plants in farmers' fields in Hapcheon, Gyeongnam Province, South Korea. Disease incidence ranged from 50 to 80% among fields. Circular- to irregular-shaped spots surrounded by yellow halos occurred frequently on the leaves of Korean raspberry plants. Brown spots became dark with wavy borders and ranged from 20 to 300 mm in diameter. Infected leaves became chlorotic, blighted, and eventually died. Fungal hyphae covered the lesions with abundant conidia and conidiophores. Fresh leaf specimens were collected from infected plants and the putative causal pathogen was isolated onto potato dextrose agar (PDA). A total of 30 isolates of the fungus were collected from diseased plants collected in the field. Fungal colonies were gray to brown on PDA. Colonies formed conidia, 38 to 210 × 8 to 20 µm, which were solitary or catenary, obclavate to cylindrical, smooth, straight or curved, and subhyaline to pale brown or brown. Conidiophores, 98 to 840 × 4 to 12 µm, were slightly or conspicuously swollen at apex, single, simple, straight or slightly flexuous, pale to midbrown, smooth, septate, thick, monotretic, and determinate or in tufts. Morphological characteristics of the fungal specimens were similar to descriptions of Corynespora cassiicola (1). A representative isolate of the pathogen was used to inoculate leaves of Korean raspberry plants for pathogenicity testing. Five leaves of a 3-month-old potted plant were sprayed with a suspension of conidia in water. Conidia were harvested from PDA cultures and adjusted to 2 × 104 conidia/ml with a hemocytometer. Five leaves sprayed with sterile distilled water served as controls. Inoculated plants were placed in a humid chamber with 100% relative humidity at 30°C for 24 h and then moved to a greenhouse. Symptoms similar to those observed in the farmers' fields developed on the inoculated leaves within 12 days, whereas the controls remained asymptomatic. The causal fungus was reisolated from the lesions of inoculated plants to satisfy Koch's postulates. To confirm the identity of the fungus, the complete internal transcribed spacer (ITS) rDNA region was amplified and sequenced (3). Amplification of the ITS region generated a 559-bp sequence (GenBank Accession No. JQ340026) with 100% similarity to sequences of C. cassiicola in GenBank (Accession No. GU138988) causing leaf spot on cassava (2). Based on the symptoms, morphological characteristics, pathogenicity, and molecular identification, this fungus was identified as C. cassiicola (1). To our knowledge, this is the first report of leaf spot caused by C. cassiicola on Korean raspberry. The recent occurrence of leaf spot on Korean raspberry suggests that C. cassiicola is spreading widely and posing a serious threat to these plants in Korea. References: (1) M. B. Ellis et al. No. 303 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Surrey, Kew, UK, 1971. (2) X.-B. Liu et al. Plant Dis. 94:916, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, Inc., New York, 1990.

Matrix effects on proficiency testing materials. Impact on accuracy of cholesterol measurement in laboratories in the nation's largest hospital system.

The objective of this collaborative study with the Department of Veterans Affairs (VA), College of American Pathologists (CAP), and the Centers for Disease Control and Prevention (CDC) was to quantitate the matrix-induced biases of cholesterol measurements on the CAP Comprehensive Chemistry Surveys materials used in proficiency testing (PT). A total of 174 VA Medical Centers outpatient clinics and clinical laboratories participate in the VA-CDC National Cholesterol Standardization and Certification Program. This study was conducted in 112 VA laboratories that have been standardized for measuring cholesterol accurately (within +/- 3.0% of the CDC reference-method values) using fresh, unfrozen, unadulterated human serum samples. Fresh serum samples and 1990 CAP Surveys materials were sent by overnight mail, and the laboratories were asked to analyze them simultaneously in triplicate in a single analytic batch run. The results showed significant matrix-effect biases with the CAP Surveys materials with six of the eight major peer groups, despite the fact that accuracy of cholesterol measurements was maintained with fresh serum samples. The magnitude and direction (positive or negative) of the matrix-effect biases were instrument, reagent, and method specific using the following peer groups: du Pont Dimension (-8.9%); Beckman CX4, CX5, and CX7 (-5.5%); Kodak Ektachem 400, 500, and 700 (+4.4%); Instrumentation Laboratory Monarch (-3.1%); Baxter Paramax (-2.4%); Technicon SMAC and RA (+1.3%); Hitachi/BMD 704 through 747 (+0.4%); and Abbott Spectrum (-0.3%). The CAP PT materials used currently do not behave in a manner identical to fresh human serum when measuring cholesterol on many, but not all, analytic systems. The observed biases due to "matrix effects" with PT materials will cause incorrect conclusions about the accuracy of many laboratory procedures performed on fresh patient specimens. This matrix-effect phenomenon will severely hamper interlaboratory accuracy transfer, standardization efforts, and monitoring performance of a laboratory's testing accuracy with the use of the current survey materials used in PT programs. Collaborative efforts are needed to (1) improve PT fluids to analytically behave more like fresh, human serum; (2) improve instrument design and reagent formulation; and (3) select methods and methodologic parameters that are more "robust" and less sensitive to the exact character of processed calibrators, quality control, and PT materials.

Renal osteodystrophy in pre-dialysis patients: ethnic difference?

The purpose of the present study is to investigate whether an ethnic difference exists in the incidence of renal osteodystrophy between Asian and Western countries in end-stage renal disease (ESRD) patients. We evaluated bone histology in 58 pre-dialysis patients (28 male, 30 female; mean age: 47.7 years). All patients had bone biopsies with quantitative histomorphometry and serological parameters such as intact PTH, osteocalcin, total alkaline phosphatase, and basal and deferoxamine-stimulated serum aluminum levels. We observed that 91.4% of all evaluated patients showed renal osteodystrophy before the start of dialytic therapy. Mild osteitis fibrosa were observed in 21 patients (36.2%), severe osteitis fibrosa in 5 patients (8.6%), mixed lesions in 7 patients (12.1%), osteomalacia in 6 patients (10.3%), aplastic bone disease in 14 patients (24.1%), and normal bone in 5 patients (8.6%). Among the bone histomorphometric parameters, fibrosis area rate (%) showed the best correlation with intact PTH, and osteocalcin and osteoid area rate (%) with total alkaline phosphatase. Aluminum-related bone disease was not observed. Among patients with aplastic bone disease, only 14.3% showed aluminum deposition of any significance (5% < stainable bone surface aluminum < 25%). In the diabetic patients, aplastic bone disease was most common, but no case was related to aluminum intoxication. In conclusion, the distribution of renal osteodystrophy in our study was different from that of Western countries in pre-dialysis patients. Our patients tended to have more mild-form osteitis fibrosa and normal findings, and less severe-form osteitis fibrosa and aplastic bone disease. Aluminum-related bone disease was not observed.

Study of drugs affecting cholesterol-induced atherosclerosis in rabbits.

1. Effects of various drugs on cholesterol-induced atherosclerosis in rabbits during the progression phase have been studied. The drugs tested are antimetabolites (mercaptopurine, hydroxyurea), surface active agents (sodiumdodecyl sulfate), inhibitor of adrenocoritcal steroid synthesis (o, p'-DDD), lysosome stablizers (chloroquine, acetylsalicylic acid) with antihistaminic (chlorpheniramine) and cholesterol binder (nystatin). 2. Mercaptopurine treatment showed marekd reduction in both atherosclerotic lesions and cholesterol concentrations of the serum and aorta. 3. Hydroxyurea reduced both the aortic cholesterol concentration and the lesions, but the serum cholesterol concentration remained high. 4. Sodiumdodecyl sulfate and o, o'-DDD showed slight inhibition of the development of atherosclerosis. 5. Pyridinocarbamate showed a slight beneficial effect on the prevention of atherosclerosis only when it was administered prior to the meal. 6. Nystatin, chloroquine and acetylsalicylic acid + chlorpheniramine showed little effect.

Systematic analysis of the effects of hepatocyte transplantation on rats with acute liver failure.

BACKGROUND/AIMS: Acute liver failure either after liver resection or as part of underlying liver disease is still associated with high mortality. Hepatocyte transplantation in various forms has attracted attention recently. However, none of those reports have investigated the thorough and systematic analysis of effect of hepatocyte transplantation on acute liver failure induced by 90% hepatectomy. Therefore, we investigated systematic analysis of effect of hepatocyte transplantation on rats with acute liver failure. METHODOLOGY: Male Sprague-Dawley rats were used. Group I rats (n = 26) received intrasplenic injection of 2 x 10(7) hepatocytes in 0.3 mL Dulbecco's modified Eagle's medium (DMEM) 24 hours prior to 90% hepatectomy. Group II rats (n = 24) received intrasplenic injection of DMEM only. Twenty-two rats from group I and 20 from group II were observed for the determination of survival time. The remaining 8 (4/each group) rats were used to assess the liver function and regeneration. RESULTS: The hepatocyte bearing spleen revealed active glucose-6-phosphatase activity. In group I rats, the survival was longer and that group had more long-term survivors than those of group II controls. In group I, there was significant increase in the ratio of weight of remnant liver lobes to body weight. At 24 hours after hepatectomy, group I rats had improved biochemical parameters compared to those of group II rats. CONCLUSIONS: In rats with acute liver failure, intrasplenic hepatocyte transplantation acts as a bridge to support experimental rats from acute liver failure to liver regeneration, prolong survival in rats with acute liver failure and improve biochemical parameters.

Accuracy of the One Touch II whole blood glucose analyzer when used by analysts with diverse technical backgrounds.

BACKGROUND: The accuracy of hand-held analytical systems is dependent on the complexity of the instrument and the technical skills of the tester. We evaluated the analytical performance of the LifeScan One Touch II glucose analyzer when used by persons with various levels of technical skill and experience. METHODS: A medical technologist conducted tests on three One Touch II systems to ensure that the units were functioning properly. The technologist then trained six analysts (a physician, a registered nurse, a licensed practical nurse, a physician assistant, and two patients) in the proper use of the glucose analyzer. RESULTS: In the hands of a medical technologist, the precision of the three glucose analyzers tested was from 1.0% to 1.9% coefficient of variation (CV) and from 2.3% to 4.2% CV for the low- and high-quality control materials, respectively. The day-to-day precision (5-day period) was from 2.1% to 3.6% CV, and from 3.5% to 4.1% CV for the low- and high-quality control materials, respectively. Comparisons of the glucose values (n = 40 fresh patient serum samples) with a reference method yielded a correlation coefficient of 0.992 to 0.993, and an overall bias of -7%. Although the values obtained by the six operators were statistically different, the differences were not clinically relevant. CONCLUSIONS: Our data suggest that the One Touch II glucose analyzer is a reliable system, and that its function is not dependent on the technical skills of the operator.

Study of user preference of graphic user interface in laboratory information system.

In order to elucidate employees satisfaction levels in using graphic user interface. (GUI) in a laboratory information system (LIS), users attitudes toward GUI versus CUI. (Character User Interface) in a similar LIS were surveyed one month after implementing upgraded version of LIS in a tertiary care university hospital laboratory. The outcome of the study showed that approximately two third of users preferred to have GUI in LIS. There was no difference in preference of GUI of users whether they had previous experience with GUI or not. However the female and older employees tend to shy away from GUI. The employee productivity was improved with GUI although it takes slightly longer program loading time than that of CUI. In conclusion, the laboratory employee satisfaction was higher with GUI than CUI and their productivity was improved with GUI.

Management and transmission of DICOM files using PC to PC multicasting methodology.

The PACS system built and used in hospitals nowadays has quite significant overload on its central server because of both treatment of very large data and full management of medical images. We suggest a distributed communication and management methodology using PC to PC multicasting strategy for efficient management of medical images produced by DICOM modalities. It is absolutely necessary for reducing strict degradation of PACS system due to large size of medical images and its very high transport rates. DICOM PC to PC component is composed a service manager to execute requested queries, a communication manager to take charge of file transmission, and a DICOM manager to manage stored data and system behavior. Each manager itself is a component to search for requested file by interaction or transmit the file to other PCs. Distributed management and transformation of medical information based on PC to PC multicasting methodology will enhance performance of central server and network capacity reducing overload on them.

Toward clinical end-user computing: programmable order protocols for efficient human computer interaction.

We have developed and implemented an efficient method of managing routine patient care information as a programmable group order protocol. The purpose of protocol is to minimize a labor-intensive manual computer interaction by grouping clinically related routine orders as a single entity, thus to greatly speed up the time taken for manual entry such as keyboard stroke and/or mouse clicking. User programmability is added to facilitate insertion, deletion and update of order items to be a locally independent operation. A sequence of menu screen is also programmable when a change of standard operation is needed. Department specific order protocols are classified into four categories to improve user convenience. The degree of efficiency is measured by a number of key strokes and entry time. In most cases the time to enter order protocol with correction is found to take less than one minute with less than five key strokes. The method of order protocol entry clearly demonstrates end-user computing capability so that department specific requirements are resolved without resorting to computer department personnel. Flexibility of managing individual physician specific protocols is also beneficial enough to enhance the morale toward a hospital information system currently in use.

Survey on medical records and EHR in Asia-Pacific region: languages, purposes, IDs and regulations.

OBJECTIVES: To clarify health record background information in the Asia-Pacific region, for planning and evaluation of medical information systems. METHODS: The survey was carried out in the summer of 2009. Of the 14 APAMI (Asia-Pacific Association for Medical Informatics) delegates 12 responded which were Australia, China, Hong Kong, India, Indonesia, Japan, Korea, New Zealand, the Philippines, Singapore, Thailand, and Taiwan. RESULTS: English is used for records and education in Australia, Hong Kong, India, New Zealand, the Philippines, Singapore and Taiwan. Most of the countries/regions are British Commonwealth. Nine out of 12 delegates responded that the second purpose of medical records was for the billing of medical services. Seven out of nine responders to this question answered that the second purpose of EHR (Electronic Health Records) was healthcare cost cutting. In Singapore, a versatile resident ID is used which can be applied to a variety of uses. Seven other regions have resident IDs which are used for a varying range of purposes. Regarding healthcare ID, resident ID is simply used as healthcare ID in Hong Kong, Singapore and Thailand. In most cases, disclosure of medical data with patient's name identified is allowed only for the purpose of disease control within a legal framework and for disclosure to the patient and referred doctors. Secondary use of medical information with the patient's identification anonymized is usually allowed in particular cases for specific purposes. CONCLUSION: This survey on the health record background information has yielded the above mentioned results. This information contributes to the planning and evaluation of medical information systems in the Asia-Pacific region.

Effect of regular exercise on senile dementia patients.

Dementia population worldwide is considerable in elderly people. Exercise regulates the brain function, but the mechanism by which it does so is unknown. The effect of regular exercise on cognitive function and exercise capacity in senile dementia patients was investigated. Thirty female patients with senile dementia who participated in the study were divided into two groups: the exercise group (EG, n = 15) and the control group (CG, n = 15). The exercise group completed a regular exercise program, and their cognitive function, activities of daily living and exercise capacity levels were evaluated at baseline, 6 months and after 12 months. Subjects exercised 30 - 60 minutes per day, 2 - 3 times per week for 12 months. Mini-mental state examination (MMSE) (pre: 14.53 +/- 5.34, post: 17.47 +/- 6.90) and ADL (pre: 14.40 +/- 5.32, post: 17.53 +/- 5.46) scores were significantly enhanced in the exercise group with senile dementia, compared to those in the control group. Exercise capacities such as cardiopulmonary function (pre: 128.47 +/- 55.43, post: 184.40 +/- 41.16), muscle strength (pre: 10.07 +/- 3.61, post: 13.7 +/- 3.90), muscular endurance (pre: 8.13 +/- 4.45, post: 12.13 +/- 5.14), flexibility (- 1.53 +/- .30, post: 2.20 +/- .70, balance (pre: 1.73 +/- .28, post: 1.20 +/- .77), and agility (pre: 21.80 +/- 3.24, post: 10.87 +/- 2.99) also increased in the exercise group. Our findings showed that regular exercise can enhance cognitive and functional activity scores in dementia patients, suggesting that senile dementia may improve by participating in a regular exercise program.

Accurate measurement of serum total cholesterol: the need for standardization.

Heightened awareness of the importance of cholesterol and heart disease has increased cholesterol testing in the United States. The demand for reliable cholesterol measurements has become a focal concern of the patient as well as the clinician. This paper covers the major analytical and preanalytical issues and factors that can affect the reliability of cholesterol results. We discuss factors that lead to impression and inaccuracy; solutions for some of the major problems; resources and techniques to help standardize cholesterol measurement; and preanalytical issues that can affect cholesterol results--i.e., patient preparation; collection, processing, storage and proper analysis of the specimen; biological and seasonal variations; age and gender; diet; alcohol consumption; weight changes; exercise; primary diseases; and infections and trauma. Many of these can be controlled by the physician, resulting in more reliable cholesterol readings under stable metabolic conditions. Accurate values will help to classify the patient's coronary heart disease risk, define appropriate treatment strategies, and simplify monitoring of dietary and/or drug intervention.

Effect of glycine on retroperitoneal and intraperitoneal organs in the rat model.

Twenty Sprague Dawley rats were administered various doses of 1.5% amino acetic acid (glycine), lactated Ringer's, and water, both intravenously and retroperitoneally, in an attempt to recreate the post-transurethral resection syndrome in a rat model. The kidneys, liver, and pancreas were harvested 6 hours after exposure and examined pathologically. Water and lactated Ringer's had no histologic effect on these organs. Glycine was found to have a toxic effect on the kidneys and liver and this effect was dose related. Based on these results, it is postulated that glycine toxicity may play a significant role as a causative factor in producing the post-transurethral resection syndrome.

Hyperprolactinaemia and verapamil: prevalence and potential association with hypogonadism in men.

OBJECTIVE: Verapamil has been associated with hyperprolactinaemia, but there have been no population-based studies. Our objective was to determine the prevalence and degree of hyperprolactinaemia associated with verapamil in the clinical setting. DESIGN: Observation with cross-sectional and longitudinal components in the setting of an urban teaching hospital and its satellite out-patient clinics. PATIENTS: Male out-patients excluding those taking other drugs known to raise PRL, renal failure and known primary hypothyroidism (1265 eligible subjects). Control subjects were drawn from eligible out-patients not taking verapamil. MEASUREMENTS: Serum PRL levels, frequency of persistent hyperprolactinaemia and total testosterone levels. RESULTS: Prolactin levels were obtained in 449 subjects on verapamil (35.5% response rate) and 166 controls. The proportions of individuals with hyperprolactinaemia (PRL > 460 mU/l) were 0.085 and 0.030 in the verapamil and control groups, respectively (P = 0.012, X2-test). The mean (+/- SD) serum PRL levels were 267 +/- 205 and 203 +/- 118 mU/l in the verapamil and control groups, respectively (P < 0.001, independent t-test). Of the 38 patients with previously determined elevated PRL levels, follow-up data were obtained in 25 (65.8%); one was found to have a pituitary adenoma and was excluded from the analysis. Fifteen of the 24 were still on verapamil (Group 1) and 14 (93.3%) continued to be hyperprolactinaemic. In 9 patients verapamil had been discontinued (Group 2) and all had normal PRL levels. Continued verapamil use was associated with persistent hyperprolactinaemia (odds ratio > 120, P < 0.00001). The mean +/- SD serum testosterone levels at follow-up were significantly lower in Group 1 (6.16 +/- 2.52 nmol/l) than in Group 2 (9.42 +/- 3.92 nmol/l, P = 0.029, independent t-test). CONCLUSIONS: The prevalence of hyperprolactinaemia associated with verapamil use in this study of male out-patients was 8.5% (95% CI 5.9-11.1%). The persistence of hyperprolactinaemia when verapamil was continued (Group 1) and the return to normal PRL levels when verapamil was discontinued (Group 2) confirm verapamil's causal role in the development of hyperprolactinaemia. While low testosterone levels were common in both groups, testosterone levels were lower in patients on verapamil. Our data suggest that screening for hyperprolactinaemia should be considered in male patients taking verapamil.

A hyperthermophilic sulfur-reducing archaebacterium, Thermococcus sp. DT1331, isolated from a deep-sea hydrothermal vent.

A hyperthermophilic archaebacterium was isolated from a deep-sea black smoker chimney (depth, 760 m) at the Minami-ensei Knoll (28 degrees 23'N, 127 degrees 38'E). The strain, designated DT1331, was a coccoid shaped bacterium about 0.5 to 1.0 microns in diameter. The cells were surrounded by a cell envelope. The temperature for growth was between 55 degrees C and 93 degrees C with an optimum 80 degrees C. The growth occurred from pH 4.5 to 8.5 and the optimum pH was 6.0. DT1331 required 1% to 5% NaCl for growth and cell lysis was observed below 1% NaCl concentration. The strain was an anaerobic chemoorganotroph requiring elemental sulfur obligately. Organic substrates used included tryptone, peptone, soytone, casein, gelatin, and yeast extract. Under the optimal conditions, DT1331 had a generation time of 50 min and could reach densities of about 1.5 x 10(8) cells/ml. DT1331 was resistant to ampicillin, chloramphenicol, erythromycin, kanamycin, streptomycin, and tetracycline, which was one of the common characteristics of archaebacteria. The G+C content of DT1331 was 52.3 mol%. Analysis of the 16S rRNA gene by restriction enzymes coincided with those of Thermococcus celer, indicating that this strain belonged to the genus Thermococcus.