Genome-wide association study of lifetime cannabis use based on a large meta-analytic sample of 32 330 subjects from the International Cannabis Consortium.

Cannabis is the most widely produced and consumed illicit psychoactive substance worldwide. Occasional cannabis use can progress to frequent use, abuse and dependence with all known adverse physical, psychological and social consequences. Individual differences in cannabis initiation are heritable (40-48%). The International Cannabis Consortium was established with the aim to identify genetic risk variants of cannabis use. We conducted a meta-analysis of genome-wide association data of 13 cohorts (N=32 330) and four replication samples (N=5627). In addition, we performed a gene-based test of association, estimated single-nucleotide polymorphism (SNP)-based heritability and explored the genetic correlation between lifetime cannabis use and cigarette use using LD score regression. No individual SNPs reached genome-wide significance. Nonetheless, gene-based tests identified four genes significantly associated with lifetime cannabis use: NCAM1, CADM2, SCOC and KCNT2. Previous studies reported associations of NCAM1 with cigarette smoking and other substance use, and those of CADM2 with body mass index, processing speed and autism disorders, which are phenotypes previously reported to be associated with cannabis use. Furthermore, we showed that, combined across the genome, all common SNPs explained 13-20% (P<0.001) of the liability of lifetime cannabis use. Finally, there was a strong genetic correlation (rg=0.83; P=1.85 × 10(-8)) between lifetime cannabis use and lifetime cigarette smoking implying that the SNP effect sizes of the two traits are highly correlated. This is the largest meta-analysis of cannabis GWA studies to date, revealing important new insights into the genetic pathways of lifetime cannabis use. Future functional studies should explore the impact of the identified genes on the biological mechanisms of cannabis use.

Logical analysis of the budding yeast cell cycle.

The budding yeast Saccharomyces cerevisiae is a model organism that is commonly used to investigate control of the eukaryotic cell cycle. Moreover, because of the extensive experimental data on wild type and mutant phenotypes, it is also particularly suitable for mathematical modelling and analysis. Here, I present a new Boolean model of the budding yeast cell cycle. This model is consistent with a wide range of wild type and mutant phenotypes and shows remarkable robustness against perturbations, both to reaction times and the states of component genes/proteins. Because of its simple logical nature, the model is suitable for sub-network analysis, which can be used to identify a four node core regulatory circuit underlying cell cycle regulation. Sub-network analysis can also be used to identify key sub-dynamics that are essential for viable cell cycle control, as well as identifying the sub-dynamics that are most variable between different mutants.

Variation in energy expenditure among black-legged kittiwakes: effects of activity-specific metabolic rates and activity budgets.

We sought to determine the effect of variation in time-activity budgets (TABs) and foraging behavior on energy expenditure rates of parent black-legged kittiwakes (Rissa tridactyla). We quantified TABs using direct observations of radio-tagged adults and simultaneously measured field metabolic rates (FMR) of these same individuals (n=20) using the doubly labeled water technique. Estimated metabolic rates of kittiwakes attending their brood at the nest or loafing near the colony were similar (ca. 1.3 x basal metabolic rate [BMR]), although loafing during foraging trips was more costly (2.9 x BMR). Metabolic rates during commuting flight (7.3 x BMR) and prey-searching flight (6.2 x BMR) were similar, while metabolic rates during plunge diving were much higher (ca. 47 x BMR). The proportion of the measurement interval spent foraging had a positive effect on FMR (R2=0.68), while the combined proportion of time engaged in nest attendance and loafing near the colony had a negative effect on FMR (R2=0.72). Thus, more than two-thirds of the variation in kittiwake FMR could be explained by the allocation of time among various activities. The high energetic cost of plunge diving relative to straight flight and searching flight suggests that kittiwakes can optimize their foraging strategy under conditions of low food availability by commuting long distances to feed in areas where gross foraging efficiency is high.

An evaluation of marine bird population trends following the Exxon Valdez oil spill, Prince William Sound, Alaska.

We examined post-spill trends (1989-1998) of marine bird populations in Prince William Sound (PWS) following the Exxon Valdez oil spill (EVOS) to evaluate recovery of injured taxa. Two criteria were employed. First, we examined population trends of injured taxa only in the oiled area of PWS using regression models. Second, we examined population trends of injured taxa in the oiled area relative to the unoiled area using homogeneity of the slopes tests. We considered a population recovering if there was a positive trend using either criteria. We considered a population not recovering if there was no trend using either criteria or a negative trend in the oiled area. A significant negative trend in the oiled area relative to the unoiled area was considered a continuing and increasing effect. Most taxa for which injury was previously demonstrated were not recovering and some taxa showed evidence of increasing effects nine years after the oil spill. Four taxa (loons Gavia spp, Harlequin Duck Histrionicus histrionicus, Bufflehead Bucephala spp, and North-western Crow Corvus caurinus) showed weak to very weak evidence of recovery. None of these taxa showed positive trends in both winter and summer. Nine taxa (grebes Podiceps spp, cormorants Phalacrocorax spp, Black Oystercatcher Haematopus bachmani, Mew Gull Larus canus, Glaucous-winged Gull Larus glaucescens, terns Sterna spp, murres Uria spp, Pigeon Guillemot Cepphus columba, and murrelets Brachyramphus spp) showed no evidence of recovery during summer or winter. Four taxa (scoters Melanitta spp, mergansers Mergus spp, goldeneyes Bucephala spp, and Black-legged Kittiwaka Rissa tridactyla) showed evidence of continuing, increasing effects. We showed evidence of slow recovery, lack of recovery, and divergent population trends in many taxa which utilize shoreline and nearshore habitats where oil is likely to persist. Potential lingering spill effects and natural variability appear to be acting in concert in delaying recovery of many PWS bird populations.

Cytologic diagnosis of malignant mesothelioma, with particular emphasis on the epithelial noncohesive cell type.

The effusion cytologies from 21 cases of malignant mesothelioma (MM) (15 pleural, 6 peritoneal) diagnosed at the Indiana University Medical Center during 1990-1997 were reviewed. Using the classification of Tao (Acta Cytol 1979;23:209-213), 13 cases of MM were of the epithelial cohesive cell type and 8 were of the epithelial noncohesive cell type. While the epithelial cohesive cell type has been discussed in the literature, the epithelial noncohesive cell type has not. The cytomorphologic features for both types are presented with particular emphasis on the noncohesive cell type. The differential diagnosis and use of ancillary confirmatory laboratory tests are briefly discussed. Because of its resemblance to florid reactive mesothelial hyperplasia and the general lack of awareness of the existence of the single-cell pattern of mesothelioma, this diagnosis can often be missed.

Atrial natriuretic peptide in preeclampsia: metabolic clearance, sodium excretion and renal hemodynamics.

To further elucidate the role of atrial natriuretic peptide (ANP) in preeclampsia, its metabolic clearance (MCRANP) was determined concomitantly with its effects on sodium excretion (UNa), glomerular filtration rate (GFR), and effective renal plasma flow (ERPF). Ten untreated preeclamptic primigravidae (PET) were studied at 29-37 wk gestation and again 4 mo postpartum (PP). Basal plasma concentration of ANP was significantly increased in PET compared with PP (14.8 +/- 1.9 vs. 4.1 +/- 0.5 pmol/l, respectively; P < 0.0001). MCRANP in PET and PP was 5.0 +/- 0.8 and 4.9 +/- 0.5 l/min [not significant (NS)], respectively. In PET, infusion of ANP produced (basal vs. ANP) a natriuresis (UNa 0.14 +/- 0.02 vs. 0.28 +/- 0.04 mmol/min, P < 0.001) and an increase in GFR (97 +/- 7 vs. 106 +/- 8 ml/min, P < 0.05), with ERPF unchanged (609 +/- 24 vs. 634 +/- 29 ml/min, NS). In PP, ANP infusion also produced a natriuresis (UNa 0.20 +/- 0.02 vs. 0.25 +/- 0.02 mmol/min, P = 0.01), no significant change in GFR (109 +/- 7 vs. 102 +/- 4 ml/min), and a significant reduction in ERPF (514 +/- 22 vs. 409 +/- 18 ml/min, P < 0.0001). Analysis of variance demonstrated a greater natriuretic effect of ANP in PET compared with PP (P < 0.05), similarly a significant difference in the effect of ANP on ERPF (P < 0.01) and GFR (P < 0.05) was seen but not on filtration fraction (P = 0.35).

The metabolic clearance of atrial natriuretic peptide during human pregnancy.

OBJECTIVE: Our purpose was to determine whether human pregnancy alters the metabolic clearance and natriuretic effect of atrial natriuretic peptide. STUDY DESIGN: The metabolic clearance rate of atrial natriuretic peptide (ANP 99-126) was measured serially in nine normotensive primigravid women studied in early and late pregnancy and again 4 months post partum (nonpregnant). Metabolic clearance of atrial natriuretic peptide was determined by use of a two-tier constant infusion technique (6 and 12 ng/kg/min, respectively). Sodium excretion was determined from 30-minute urine collections taken before and during infusion of atrial natriuretic peptide at both 6 and 12 ng/kg/min. RESULTS: Basal plasma atrial natriuretic peptide levels increased with gestation: in early pregnancy 18.0 +/- 2.7 pg/ml, in late pregnancy 22.6 +/- 4.2 pg/ml, and post partum 19.5 +/- 3.6 pg/ml. Infusion of atrial natriuretic peptide at 6 and 12 ng/kg/min produced two distinct physiologic plasma levels of atrial natriuretic peptide. The metabolic clearance rates for nonpregnant women and those in early and late pregnancy at 6 and 12 ng/kg/min, respectively, were 3.4 +/- 0.4 and 2.9 +/- 0.4 L/min at plasma atrial natriuretic peptide levels of 86.2 +/- 13.2 and 179.8 +/- 42.5 pg/ml, respectively, 4.3 +/- 0.5 and 4.3 +/- 0.5 L/min at plasma atrial natriuretic peptide levels of 61.1 +/- 4.9 and 131 +/- 20.9 pg/ml (p < 0.01, nonpregnant vs early pregnancy), and 3.8 +/- 0.6 and 3.8 +/- 0.5 L/min at plasma atrial natriuretic peptide levels of 72 +/- 8.0 and 136 +/- 18.3 pg/ml (p < 0.05, nonpregnant vs late pregnancy), respectively. Infusion of atrial natriuretic peptide produced natriuresis in both pregnant and nonpregnant states; sodium excretion (basal to atrial natriuretic peptide infusion at 12 ng/kg/min) increased from 133 +/- 19 to 207 +/- 18 mumol/min, 129 +/- 21 to 374 +/- 35 mumol/min, and 128 +/- 20 to 221 +/- 33 mumol/min in nonpregnant women and those in early and late pregnancy, respectively. CONCLUSIONS: The metabolic clearance of atrial natriuretic peptide increased by 16 weeks' gestation and remained elevated thereafter. There appears to be no attenuation of the natriuretic effect of infused atrial natriuretic peptide in normotensive human pregnancy.

Effect of atrial natriuretic peptide on renal hemodynamics and sodium excretion during human pregnancy.

The effect of infused atrial natriuretic peptide (ANP) on sodium excretion (UNa), glomerular filtration rate (GFR), and effective renal plasma flow (ERPF) was studied in 12 normotensive primigravidae at 32 wk gestation [late pregnancy (LP)] and again 4 mo postpartum [nonpregnant (NP)]. Three 20-min steady-state (renal) clearances of inulin and p-aminohippurate were used to measure GFR and ERPF, respectively, before and after infusion of ANP at 2 pmol.kg-1.min-1. Basal plasma ANP (pANP) was increased in LP compared with NP [7.8 +/- 0.6 vs. 3.3 +/- 0.4 pmol/l (P < 0.0001), respectively]. In LP, infusion of ANP increased pANP from 7.8 +/- 0.6 to 21.8 +/- 1.4 pmol/l (P < 0.00001), which produced a natriuresis [UNa of 0.18 +/- 0.02 vs. 0.25 +/- 0.03 mmol/min (P = 0.03), respectively], with no change in GFR (153 +/- 13 vs. 142 +/- 8 ml/min, P = 0.16) but a significant reduction in ERPF (766 +/- 52 vs. 660 +/- 31 ml/min, P = 0.002). In NP, ANP infusion increased pANP from 3.3 +/- 0.4 to 27.7 +/- 2.5 pmol/l (P < 0.00001), which produced no significant natriuresis [UNa of 0.22 +/- 0.07 vs. 0.26 +/- 0.09 mmol/min (P = 0.15), respectively] and no change in GFR (87 +/- 3 vs. 89 +/- 3 ml/min), but again a reduction in ERPF (486 +/- 17 vs. 414 +/- 9 ml/min, P < 0.001).

Thirst induced by a suckling episode during breast feeding and relation with plasma vasopressin, oxytocin and osmoregulation.

OBJECTIVE: There have been anecdotal reports of increased maternal thirst during breast feeding, but the physiological mechanisms remain obscure. We have assessed and quantified the stimulation of maternal thirst during breast feeding (suckling), and correlated the changes with plasma oxytocin and vasopressin levels. DESIGN: A within subject design was used with each subject acting as her own control. Each subject participated in a suckling period and a control non-suckling period the order of which was counterbalanced. SUBJECTS: Ten healthy breastfeeding women ranging from 28 to 52 days post partum. MEASUREMENTS: Thirst (assessed by a visual analogue scale), plasma vasopressin, plasma oxytocin (pOT), plasma osmolality and haematocrit, blood pressure, volume of milk transferred to baby, volume of water drunk immediately following each experimental period. RESULTS: Thirst increased significantly more over a suckling period than over a comparable control period (P = 0.013). The peak level of thirst correlated with the volume of water consumed to satiety following the suckling period (r = 0.7, P = 0.024). As expected, pOT levels rose significantly in response to suckling (P < 0.001). Six women demonstrated a close relation between thirst and oxytocin response during the suckling period. Despite significant changes in thirst during suckling there was no increase in plasma vasopressin or in osmoregulatory parameters. CONCLUSIONS: This study confirms that suckling is a potent stimulus to thirst in the mother, and is not associated with vasopressin release or dependent on any measurable alterations in osmoregulation. What actually stimulates thirst during breast feeding remains unknown, but there are two potential explanations for these observations: (1) suckling sends nerve impulses to the paraventricular and supraoptic nuclei in the hypothalamus which may have afferents within the central nervous system which stimulates a thirst response simultaneous with oxytocin release; (ii) a learned anticipation of thirst may be occurring in a situation associated with expectant fluid loss to preserve homeostasis.

The severe perinatal form of autosomal recessive polycystic kidney disease maps to chromosome 6p21.1-p12: implications for genetic counseling.

Autosomal recessive polycystic kidney disease (ARPKD) is a one of the most common hereditary renal cystic diseases in children. Its clinical spectrum is widely variable with most cases presenting in infancy. Most affected neonates die within the first few hours of life. At present, prenatal diagnosis relies on fetal sonography, which is often imprecise in detecting even the severe form of the disease. Recently, in a cohort of families with mostly milder ARPKD phenotypes, an ARPKD locus was mapped to a 13-cM region of chromosome 6p21-cen. To determine whether severe perinatal ARPKD also maps to chromosome 6p, we have analyzed the segregation of seven microsatellite markers from the ARPKD interval in 22 families with the severe phenotype. In the majority of the affected infants, ARPKD was documented by histopathology. Our data confirm linkage and refine the ARPKD region to a 3.8-cM interval, delimited by the markers D6S465/D6S427/D6S436/D6S272 and D6S466. Taken together, these results suggest that, despite the wide variability in clinical phenotypes, there is a single ARPKD gene. These linkage data and the absence of genetic heterogeneity in all families tested to date have important implications for DNA-based prenatal diagnoses as well as for the isolation of the ARPKD gene.

Evaluation of transvaginal sonography combined with a urinary luteinizing hormone monitor in timing donor insemination.

There remains controversy as to the most accurate method of predicting ovulation for the timing of donor insemination. In a prospective study based at a tertiary referral donor insemination clinic we have assessed the ability of a urinary luteinizing hormone (LH) kit combined with vaginal ultrasonography to predict ovulation for donor insemination. A total of 25 natural cycles were monitored from the first day of the LH surge (detected by the urinary kit). Daily transvaginal sonographic measurement of follicular size and endometrial thickness were used to predict ovulation; monitoring continued until post-ovulatory changes were seen. Transvaginal sonography combined with a urinary LH kit successfully predicted all those women who ovulated (20/25) and detected unfavourable conditions for insemination in the remaining 20% (5/25). In conclusion, where available transvaginal sonography combined with a urinary LH kit should be the method of choice for timing donor insemination.

A simple alternative to parenteral oxytocics for the third stage of labor.

OBJECTIVE: To determine the effect of nipple stimulation on uterine activity during the third stage of labor. METHODS: Randomized controlled study comparing; (i) 15 min of nipple stimulation (n = 6), (ii) routine syntometrine injection (n = 3), (iii) no action/control (N = 5). Uterine activity was continuously measured using the placenta as an in-situ hydrostatic bag connected to a pressure transducer. RESULTS: Compared to controls uterine pressure was higher during nipple stimulation (103 mmHg vs. 70.8 mmHg, P = 0.04). The duration of the third stage and blood loss tended to be reduced with nipple stimulation compared to controls (20.3 vs. 12.3 min) and (257 vs. 166 ml) respectively but was not significant. Similar differences were observed between syntometrine and control groups. CONCLUSIONS: For women in developing countries where parenteral oxytocics are not available, nipple stimulation might reduce the incidence of postpartum hemorrhage. A larger trial now seems warranted.

Ubombo and the site of David Bruce's discovery of Trypanosoma brucei.

The site and remains believed to be those of the camp where Sir David Bruce and his wife Mary worked between 1894 and 1897, and where Bruce discovered the causative agent of nagana and its transmission by the tsetse fly, have recently been discovered at the small village of Ubombo in northern KwaZulu (Zululand), South Africa. The site where these remnants were found fits the meagre, albeit significant, information presented by Bruce in his writings on the location of the camp.