The Caenorhabditis elegans GARP complex contains the conserved Vps51 subunit and is required to maintain lysosomal morphology.

In yeast the Golgi-associated retrograde protein (GARP) complex is required for tethering of endosome-derived transport vesicles to the late Golgi. It consists of four subunits--Vps51p, Vps52p, Vps53p, and Vps54p--and shares similarities with other multimeric tethering complexes, such as the conserved oligomeric Golgi (COG) and the exocyst complex. Here we report the functional characterization of the GARP complex in the nematode Caenorhabditis elegans. Furthermore, we identified the C. elegans Vps51 subunit, which is conserved in all eukaryotes. GARP mutants are viable but show lysosomal defects. We show that GARP subunits bind specific sets of Golgi SNAREs within the yeast two-hybrid system. This suggests that the C. elegans GARP complex also facilitates tethering as well as SNARE complex assembly at the Golgi. The GARP and COG tethering complexes may have overlapping functions for retrograde endosome-to-Golgi retrieval, since loss of both complexes leads to a synthetic lethal phenotype.

Influence of age and sex in exhaled breath samples investigated by means of infrared laser absorption spectroscopy.

Breath gas analysis provides insight into human metabolism of healthy and ill individuals. As an innovative and non-invasive method, it opens up options to improve diagnostics, monitoring and treatment decisions. Mid-infrared laser absorption spectroscopy is utilized to detect CH(4), H(2)O, CO(2), NH(3) and CH(3)OH in exhaled human breath. An off-line approach using breath sampling by means of Tedlar bags is applied. The breath gas samples are measured within the population-based epidemiological Study of Health in Pomerania (SHIP-TREND) performed at the University of Greifswald. The study covers about 5000 adult subjects aged 20-79 years within 3 years. Besides breath gas analysis many other examinations are conducted. It is expected to find associations between distinct concentration levels of species in the exhaled breath and diseases assessed in this study. The study will establish reference values for exhaled breath components and serve as background population for case-control studies. In the long run, morbidity and mortality follow-ups will be conducted, which will answer the question whether end-expiratory breath gas components predict future diseases and death. As first results, we present data from 45 dialysis patients (23 males, 22 females) which were recruited in a preliminary study in preparation for SHIP-TREND.

Incidence, pathology and outcome of gynaecological cancer in patients under the age of 21 years in South-west England 1995-2004: comparison of data from regional, national and international registries.

We analysed the incidence, tumour types, management and outcome of gynaecological cancer diagnosed from 1995-2004 in females <21 years in south-west England. Data from the South West Cancer Intelligence Service were compared with those from regional and national registries. A total of 63 patients had gynaecological malignancies: 49 ovarian; nine cervical; the remainder vaginal, uterine or pelvic. The median age was 16 years. Germ cell tumours (26) and carcinomas (6) were the commonest primary ovarian and cervical tumours respectively. Most patients had fertility-sparing procedures. Only seven required re-operation. Information about chemoradiotherapy was incomplete. Four deaths occurred. All patients were followed >3 years and 68% >5 years, with 94% survival to date. Fertility preservation did not impair survival. Mortality is an inadequate indicator of outcome; cancer registries should record information on fertility and pregnancy outcomes, second tumours and long-term treatment-related complications. Improved management requires greater centralised assessment of histology, follow-up and adjuvant treatment.

Paternal insulin resistance and its association with umbilical cord insulin concentrations.

AIMS/HYPOTHESIS: Fetal growth is influenced by genetic factors as well as the intra-uterine environment. We hypothesised that some genetic factors may alter fetal insulin secretion and insulin action. SUBJECTS, MATERIALS AND METHODS: To assess this, we analysed plasma insulin concentration in umbilical cord blood from 644 normal, term, UK Caucasian deliveries from the Exeter Family Study of Childhood Health. We tested for associations between cord insulin and each of parental anthropometry, fasting glucose, insulin and lipids. RESULTS: As expected, cord insulin concentrations correlated with all measures of birth size (weight, length, head and arm circumferences, sum of skinfold thicknesses, ponderal index: r=0.16-0.4, p<0.01 for all) and maternal BMI (r=0.11, p=0.005), maternal glucose (r=0.25, p<0.001) and maternal insulin resistance (r=0.23, p<0.001). Paternal fasting insulin and insulin resistance were correlated with cord insulin (r=0.15, p=0.006; r=0.13, p=0.001, respectively), and this was independent of paternal BMI. Multiple linear regression analysis revealed paternal insulin resistance to be a predictor of cord insulin concentrations, independently of maternal factors. CONCLUSION: Our results show an independent relationship between paternal insulin resistance and cord insulin concentrations. This is consistent with heritability of insulin resistance from father to offspring and a compensatory increase in fetal insulin secretion, the latter occurring pre-natally before the homeostatic feedback loop between glucose and insulin is established.

Ovarian cancer death reduction for women at high risk: workload implications for gynaecology services.

Ovarian cancer spreads early, presents late and is difficult to cure. Reducing death rates from ovarian malignancy has focussed on the unaffected females of families with a high chance of a mutant gene such as BRCA1 and BRCA2. We set up a familial ovarian cancer service in a district general hospital in the UK, serving a population of 330,000. The clinical genetics team acted as gatekeepers to the service. Risk assessment, ultrasound and biochemical screening and prophylactic oophorectomy were discussed. Gene testing was offered when appropriate. This study reviews the levels of activity for the first 8 years of the service. In all, 153 women were referred, of whom 34 (16%) did not have significant clinical histories. Of the 114 who fulfilled the UKFOCSS criteria for family history and age, four were quickly found to be gene mutation negative and 20 (17%) declined intervention. A total of 29 (25%) chose prophylactic oophorectomy and 61 (54%) chose ultrasound screening.

beta-cell genes and diabetes: molecular and clinical characterization of mutations in transcription factors.

beta-Cell transcription factor genes are important in the pathophysiology of the beta-cell, with mutations in hepatocyte nuclear factor (HNF)-1alpha, HNF-4alpha, insulin promoter factor (IPF)-1, HNF-1beta, and NeuroD1/BETA2, all resulting in early-onset type 2 diabetes. We assessed the relative contribution of these genes to early-onset type 2 diabetes using linkage and sequencing analysis in a cohort of 101 families (95% U.K. Caucasian). The relative distribution of the 90 families fitting maturity-onset diabetes of the young (MODY) criteria was 63% HNF-1alpha, 2% HNF-4alpha, 0% IPF-1, 1% HNF-1beta, 0% NeuroD1/ BETA2, and 20% glucokinase. We report the molecular genetic and clinical characteristics of these patients including 29 new families and 8 novel HNF-1alpha gene mutations. Mutations in the transactivation domain are more likely to be protein truncating rather than result in amino acid substitutions, suggesting that a relatively severe disruption of this domain is necessary to result in diabetes. Mutations in the different transcription factors result in clinical heterogeneity. IPF-1 mutations are associated with a higher age at diagnosis (42.7 years) than HNF-1alpha (20.4 years), HNF-1beta (24.2 years), or HNF-4alpha (26.3 years) gene mutations. Subjects with HNF-1beta mutations, in contrast to the other transcription factors, frequently present with renal disease. A comparison of age at diagnosis between subjects with different types and locations of HNF-1alpha mutations did not reveal genotype-phenotype correlations. In conclusion, mutations in transcription factors expressed in the beta-cell are the major cause of MODY, and the phenotype clearly varies with the gene that is mutated. There is little evidence to indicate that different mutations within the same gene have different phenotypes.

Adverse endothelial function and the insulin resistance syndrome.

Type 2 diabetes is characterized by impaired endothelial dependent vasodilatation which may contribute to the high prevalence of vascular disease in such patients. Although hyperglycaemia, dyslipidaemia and hypertension can all independently cause a similar defect, recent data suggest that endothelial dysfunction may be intrinsic to the insulin resistance syndrome that commonly precedes type 2 diabetes. Such abnormalities in endothelial function could represent the impact of subclinical disturbance of metabolism or alternatively the presence of a common cellular defect that influences both nitric oxide bioavailability and insulin mediated glucose disposal. Resolution of this puzzle is likely to lead to important advances in our knowledge and ultimately treatment of vascular disease.

Interactions of buspirone or gepirone with nicotine on schedule-controlled behavior of pigeons.

The primary purpose of the present study was to examine the interaction of buspirone with nicotine in pigeons responding under a fixed-ratio 30 schedule of food presentation. The hypothesis was that the dopamine D2 receptor antagonist activity of buspirone would attenuate the rate-decreasing effects of nicotine. When administered alone, buspirone (0.3-10mg/kg) and (-)-nicotine (0.1-3.0mg/kg) decreased response rates in a dose-related manner, with ED(50) values (+/-95% C.L.) of 3.0 (1.7-5.1) mg/kg and 1.0 (0.7-1.5) mg/kg, respectively. Low doses of buspirone (0.3-1.0mg/kg) did not significantly shift the nicotine dose-response function, while doses of buspirone (3.0-10mg/kg) that produced rate-decreasing effects shifted the nicotine dose-response function to the left. There was no significant statistical interaction between buspirone and nicotine indicating that the shifts in the nicotine dose-response function were parallel. The buspirone analog gepirone (0.3-10mg/kg), which like buspirone is a serotonin (5-HT(1A)) agonist, but unlike buspirone is relatively devoid of D2 antagonist activity, was also tested in combination with nicotine. Gepirone was less potent in decreasing response rates compared with buspirone, with an ED(56) value of 4.5 (3.1-6.7) mg/kg. Rate-decreasing doses of gepirone (3.0-10mg/kg) in combination with nicotine resulted in parallel shifts to the left of the nicotine dose-response function. There was no statistically significant difference between the effects of buspirone and those of gepirone on the nicotine dose-response function. Isobolograms indicated that the pharmacological interactions between buspirone or gepirone and nicotine were not different from additivity. These results suggest that the combined effects of buspirone and nicotine on schedule-controlled behavior are independent of antagonism at D2 receptors.

[Materials science studies on the soldering of different orthodontic wires]. / Werkstoffkundliche Untersuchungen zum Löten verschiedener orthodontischer Drähte.

In orthodontic technique both soldering and welding are standard methods for the application of auxiliaries and for the modification of force systems by joining wires of different cross-sections. Two cobalt-chromium alloys (Blue Elgiloy, Crozat) and an austenitic stainless steel alloy (Remanium) were soldered by an electrochemically generated hydrogen-oxygen flame forming an overlapped joint design. For characterization of the soldered joint testing procedures included microhardness tests, metallographic examination, tension-shear tests and surface analysis of the fractured joints by scanning electron microscopy. For any given soldering technique with an overlapped joint design the correct joint length is determined by the ratio t/s = 3 (t = overlapped length; s = diameter of the smaller wire).