Assessing stress physiology within a conservation breeding program for an endangered species.

Conservation breeding programs typically involve the management of individuals both in and ex situ, so it is vital to understand how the physiology of managed species changes in these environments to maximize program outcomes. The Vancouver Island marmot (VIM; Marmota vancouverensis) is one species that has been managed in a conservation breeding program to recover the critically low wild population. Previous research has shown there are differences in hair glucocorticoid concentrations for VIMs in different managed groups in the program. Therefore, we used >1000 blood samples collected since the program's inception to assess the neutrophil to lymphocyte (N:L) ratio among captive, pre-release, post-release and wild populations as another metric of stress. In situ VIM populations were found to have a significantly higher N:L ratio than ex situ populations, suggesting that the wild is a more physiologically challenging environment than managed care. Moreover, the effect of age, sex and the month of sampling on the N:L ratio were found to be different for each population. Age had the greatest magnitude of effect in the wild population, and sex was only significant in ex situ populations. This study provided previously unknown insights into the physiology of VIMs and increased post-release monitoring will be useful in the future to fully understand how physiology may be contributing to differences in survival of VIMs in the program.

Diffusion magnetic resonance imaging assessment of regional white matter maturation in preterm neonates.

PURPOSE: Diffusion magnetic resonance imaging (dMRI) studies report altered white matter (WM) development in preterm infants. Neurite orientation dispersion and density imaging (NODDI) metrics provide more realistic estimations of neurite architecture in vivo compared with standard diffusion tensor imaging (DTI) metrics. This study investigated microstructural maturation of WM in preterm neonates scanned between 25 and 45 weeks postmenstrual age (PMA) with normal neurodevelopmental outcomes at 2 years using DTI and NODDI metrics. METHODS: Thirty-one neonates (n = 17 male) with median (range) gestational age (GA) 32+1 weeks (24+2-36+4) underwent 3 T brain MRI at median (range) post menstrual age (PMA) 35+2 weeks (25+3-43+1). WM tracts (cingulum, fornix, corticospinal tract (CST), inferior longitudinal fasciculus (ILF), optic radiations) were delineated using constrained spherical deconvolution and probabilistic tractography in MRtrix3. DTI and NODDI metrics were extracted for the whole tract and cross-sections along each tract to assess regional development. RESULTS: PMA at scan positively correlated with fractional anisotropy (FA) in the CST, fornix and optic radiations and neurite density index (NDI) in the cingulum, CST and fornix and negatively correlated with mean diffusivity (MD) in all tracts. A multilinear regression model demonstrated PMA at scan influenced all diffusion measures, GA and GAxPMA at scan influenced FA, MD and NDI and gender affected NDI. Cross-sectional analyses revealed asynchronous WM maturation within and between WM tracts.). CONCLUSION: We describe normal WM maturation in preterm neonates with normal neurodevelopmental outcomes. NODDI can enhance our understanding of WM maturation compared with standard DTI metrics alone.

Early postnatal maternal trait anxiety is associated with the behavioural outcomes of children born preterm <33 weeks.

Maternal ante- and postnatal anxiety have been associated with children's socio-emotional development. Moreover, maternal anxiety has been studied as both a contributing factor and consequence of preterm birth, and children born preterm are more likely to develop behavioural problems compared to term-born controls. This study investigated the association between maternal anxiety measured soon after birth and mental health in 215 ex-preterm children, born at <33 weeks, who participated in the Evaluation of Preterm Imaging Study. Children were followed-up at a median age of 4.6 years (range 4.2-6.6), and received behavioural and cognitive evaluation. Maternal trait anxiety was assessed with the Spielberger State-Trait Anxiety Index at term corrected age. Primary outcome measures were children's Strengths and Difficulties Questionnaire (SDQ) and Social Responsiveness Scale 2 (SRS-2) scores, indicative of generalised psychopathology and autism symptomatology, respectively. IQ was assessed with the Wechsler Preschool and Primary Scales of Intelligence. The final sample, after excluding participants with missing data and multiple pregnancy (n = 75), consisted of 140 children (51.4% male). Results showed that increased maternal trait anxiety at term corrected age was associated with children's higher SDQ scores (ß = 0.25, 95% CI 0.09-0.41, p = 0.003, f2 = 0.08) and SRS-2 scores (ß = 0.15, 95% CI 0.02-0.28, p = 0.03, f2 = 0.04). Our findings indicate that children born preterm whose mothers are more anxious in the early postnatal period may show poorer mental health outcomes at pre-school age. Further research is needed to investigate preventative measures that can be offered to high-risk premature babies and their families.

MRI Findings at Term-Corrected Age and Neurodevelopmental Outcomes in a Large Cohort of Very Preterm Infants.

BACKGROUND AND PURPOSE: Brain MR imaging at term-equivalent age is a useful tool to define brain injury in preterm infants. We report pragmatic clinical radiological assessment of images from a large unselected cohort of preterm infants imaged at term and document the spectrum and frequency of acquired brain lesions and their relation to outcomes at 20 months. MATERIALS AND METHODS: Infants born at <33 weeks' gestation were recruited from South and North West London neonatal units and imaged in a single center at 3T at term-equivalent age. At 20 months' corrected age, they were invited for neurodevelopmental assessment. The frequency of acquired brain lesions and the sensitivity, specificity, and negative and positive predictive values for motor, cognitive, and language outcomes were calculated, and corpus callosal thinning and ventricular dilation were qualitatively assessed. RESULTS: Five hundred four infants underwent 3T MR imaging at term-equivalent age; 477 attended for assessment. Seventy-six percent of infants had acquired lesions, which included periventricular leukomalacia, hemorrhagic parenchymal infarction, germinal matrix-intraventricular hemorrhage, punctate white matter lesions, cerebellar hemorrhage, and subependymal cysts. All infants with periventricular leukomalacia, and 60% of those with hemorrhagic parenchymal infarction had abnormal motor outcomes. Routine 3T MR imaging of the brain at term-equivalent age in an unselected preterm population that demonstrates no focal lesion is 45% sensitive and 61% specific for normal neurodevelopment at 20 months and 17% sensitive and 94% specific for a normal motor outcome. CONCLUSIONS: Acquired brain lesions are common in preterm infants routinely imaged at term-equivalent age, but not all predict an adverse neurodevelopmental outcome.

Temporal, spatial, and management variability in the carbon footprint of New Zealand milk.

The carbon footprint of milk from year-round grazed-pasture dairy systems and its variability has had limited research. The objective of this study was to determine temporal, regional, and farm system variability in the carbon footprint of milk from New Zealand (NZ) average dairy production. Farm production and input data were collected from a national database for 2010/11 to 2017/18 across regions of NZ and weighted on relative production supplied to the major dairy cooperative Fonterra to produce an NZ-average. Total greenhouse gas emissions were calculated using a life cycle assessment methodology for the cradle-to-farm gate, covering all on- and off-farm contributing sources. The NZ-average carbon footprint of milk varied from 0.81 kg of CO2 equivalent (CO2eq)/kg of fat- and protein-corrected milk (FPCM) in 2010/11 (with widespread drought) to 0.75 to 0.78 kg of CO2eq/kg of FPCM in 2013/14 to 2017/18, with a trend for a small decrease over time. Regional variation occurred with highest carbon footprint values for the Northland region due to greatest climatic and soil limitations on pasture production. Dairy cattle diet was approximately 85% from grazed pasture with up to 15% from brought-in feeds (mainly forages and by-products). The CO2 emissions from direct fuel and electricity use constituted <2% of total CO2eq emissions, whereas enteric methane was near 70% of the total. An estimate of potential contribution from direct land use change (plantation forest to pasture) was 0.13 kg of CO2eq/kg of FPCM. This was not included because nationally there has been a net increase in forest land and a decrease in pasture land over the last 20 yr. Data used were highly representative, as evident by the same estimated carbon footprint from 368 farms (in 2017/18) from the national database compared with that from a direct survey of 7,146 farms. New Zealand-specific nitrous oxide emission factors were used, based on many validated field trials and as used in the NZ greenhouse gas inventory, resulting in an 18% lower carbon footprint than if default Intergovernmental Panel on Climate Change factors had been used. Evaluation of the upper and lower quartiles of farms based on per-cow milk production (6,044 vs. 3,542 kg of FPCM/cow) showed a 15% lower carbon footprint for the upper quartile of farms, illustrating the potential for further decrease in carbon footprint with improved farm management practices.

Early or Late Conversion From Tac-BD to Tac-BD in Renal Transplantation: When is the Right Time?

BACKGROUND: Increasing numbers of renal transplant recipients are converted from Prograf (Astellas Pharma, Tokyo, Japan) (tacrolimus twice daily [Tac-BD]) to Advagraf (Astellas) (tacrolimus once daily [Tac-QD]), but the optimal time for conversion is as yet unclear. This study aimed to investigate the correlation between the time of conversion from Tac-BD to Tac-QD after renal transplant and the dosing requirements, tacrolimus levels, renal function, and clinical outcomes. METHODS: Since September 2008, 125 renal transplant patients were converted from Tac-BD to Tac-QD and followed up for 2 years after conversion. Patients were split into early (0 to 12 months) and late (>12 months) conversion groups. Demographics, Tac-QD dose, trough levels, graft function, and patient and graft outcome were prospectively collected. RESULTS: Forty-four patients (35.2%) were converted early (3.82 ± 3.24 months), whereas 81 (64.8%) patients were converted late (77.35 ± 53.71 months). Tac-BD dose before conversion was higher in the early group (8.70 ± 6.34 vs 4.44 ± 2.15 mg) as was the initial Tac-QD dose (8.66 ± 6.20 vs 4.37 ± 2.04 mg, P < .0001), and remained higher for 18 months after conversion, as did the serum tacrolimus trough level levels. Renal function, acute rejection, and patient and graft survival were comparable between the groups. CONCLUSIONS: Patients can be safely converted to Tac-QD within the first year post-transplantation, without adverse effects on clinical outcome, despite the higher doses and tacrolimus levels for the first 18 months.

Undernutrition regulates the expression of a novel splice variant of myostatin and insulin-like growth factor 1 in ovine skeletal muscle.

Undernutrition suppresses the growth of skeletal muscles and alters the expression of insulin-like growth factor 1 (IGF1), a key mitogen, and myostatin, a potent inhibitor of myogenesis. These changes can explain, at least in part, the reduced growth of skeletal muscles in underfed lambs. We have recently identified a myostatin splice variant (MSV) that binds to and antagonizes the canonical signaling of myostatin. In the present study, we hypothesized that the expression of MSV would be reduced in conjunction with myostatin and IGF1 in response to underfeeding in skeletal muscles of sheep. Young growing ewes were fed either ad libitum or an energy-restricted diet (30% of maintenance requirements) for 28 d. This regime of underfeeding resulted in a 24% reduction in body mass (P < 0.001) and a 36% reduction in the mass of the semitendinosus muscles relative to controls (P < 0.001) by day 28. The concentrations of MSV and IGF1 messenger RNA (mRNA) were reduced (both P < 0.001), but myostatin mRNA was not altered in semitendinosus muscles. Unlike the reduced expression of mRNA, the abundance of MSV protein was increased (P < 0.05) and there was no change in the abundance of myostatin protein. Our results suggest that undernutrition for 28 d decreases the signaling of myostatin by increasing the abundance of MSV protein. Although this action may reduce the growth inhibitory activity of myostatin, it cannot prevent the loss of growth of skeletal muscles during undernutrition.

Conversion from twice-daily to once-daily tacrolimus in simultaneous pancreas-kidney transplant patients.

BACKGROUND: Data on the effectiveness of once-daily tacrolimus (Tac-QD) in simultaneous pancreas-kidney (SPK) transplant patients are limited, which is of particular concern because diabetic gastroparesis may affect absorption. The aim of this study was to evaluate the clinical impact of converting SPK patients from twice-daily (Tac-BD) to Tac-QD. METHODS: From November 2008 to August 2011, 27 SPK recipients (out of 130) were converted from Tac-BD to Tac-QD. Demographics, prescribed doses, trough levels, and creatinine, glucose, and HbA1c values were collected prospectively at the time of conversion and at 1, 2, 3, 6, and 12 months after conversion. RESULTS: The mean time from transplantation to conversion was 35.81 ± 27.31 months, with 20 patients (74.07%) converted to Tac-QD >12 months after transplantation. There were no significant differences in the tacrolimus dose and trough levels before and after conversion and at all points during the follow-up. Creatinine, glucos,e and HbA1c levels remained stable throughout. Eight patients (29.63%) with gastroparesis had clinical outcomes, drug doses, and trough levels similar to all other patients. CONCLUSIONS: Stable SPK recipients can safely be converted from Tac-BD to Tac-QD, with no clinical impact on the transplant function. Gastroparesis does not appear to influence tacrolimus dose requirements or trough levels.

Does national sharing of kidneys donated after cardiac death lead to poorer outcomes?

BACKGROUND: Kidneys donated after cardiac death (DCD) represent an increasing proportion of transplant activity. There have been concerns that wider sharing of these kidneys increases the cold ischemic time (CIT) and leads to poorer outcomes. METHODS: DCD kidney transplantation was implemented in Scotland in 2005, with each center transplanting locally donated kidneys. A national sharing scheme of DCD kidneys was introduced in 2007, whereby kidneys are shared between the 2 renal transplant centers in the country. A single national multiorgan retrieval team carries out retrievals and kidneys are shipped directly to the 2 units. Donor and recipient demographic data, cold ischemic time, and outcome data were prospectively collected and compared within each center and between centers pre- and postintroduction of the sharing policy. RESULTS: Since 2005, 152 DCD kidney transplants have been performed. Since 2007, 68 kidneys were shared between the centers. Recipient demographics were comparable before and after the introduction for the sharing scheme. The CIT was significantly higher in Glasgow (14.30 ± 3.79 hours) compared with Edinburgh (10.72 ± 2.99 hours; P < .001, one-way analysis of variance [ANOVA] prior to the introduction of the sharing scheme. Following the implementation of kidney sharing, there was no significant difference in CIT between Glasgow and Edinburgh (10.50 ± 3.34 hours vs 10.53 ± 2.71 hours). A significant reduction in the CIT in Glasgow was noted after sharing was instituted (from 14.30 ± 3.79 hours to 10.50 ± 3.34 hours, P < .001, one-way ANOVA). Patient and graft survivals, acute rejection, and delayed graft function as well as 1-year renal function were comparable in both centers before and after the introduction of the scheme. CONCLUSION: Wider sharing of DCD kidneys should be encouraged, as it does not compromise clinical outcomes. A transparent and well-established sharing agreement, with no delays in the offering of DCD kidneys, may lead to an improvement in CIT.

Fluoride in still bottled water in Australia.

BACKGROUND: This study measured the fluoride content in a range of still bottled waters available in Australia. Increasing decay rates in children have prompted speculation that bottled water consumption may be impacting on dental caries rates. International studies have demonstrated that the fluoride levels in bottled water are often negligible, however there is little information about the fluoride content of still bottled water in Australia. METHODS: One hundred different brands (300 samples) of still bottled water were obtained in Australia and tested using ion chromatography and an ion-selective electrode method. RESULTS: The tested fluoride levels ranged from <0.1 to 1.6 mg/L. Nine per cent of the samples reported fluoride levels within the range recommended for reticulated water in Australia, providing a dental health benefit. One per cent of samples recorded fluoride values greater than this range, from 1.2 to 1.6 mg/L. The majority of samples (91%) recorded fluoride levels below 0.6 mg/L. CONCLUSIONS: Better information about the fluoride levels in bottled water is of value, particularly for communities lacking access to a fluoridated drinking water supply.

Glutamate is a transmitter that mediates inhibition at the rectifying electrical motor giant synapse in the crayfish.

Spike transmission at the electrical synapse between the giant fibres (GFs) and motor giant neurone (MoG) in the crayfish can be blocked by depolarising postsynaptic chemical inhibition, which has previously been shown to be mediated in part by gamma-aminobutyric acid (GABA). The authors show that glutamate applied to the synaptic region of the MoG mimics the depolarisation of the chemical input and can also block spike transmission from the GFs. The glutamate induces an inward current mediated by a conductance increase that is 30-40% of that induced by GABA and that is blocked substantially by picrotoxin. Glutamate has no effect on the presynaptic GF, and the effects in the MoG are maintained in the presence of cadmium, indicating that the glutamate is acting directly on the MoG. Both GABA and glutamate have similar effects on the cell body, where the response reverses 10-20 mV positive to resting potential, is dependent on chloride concentration, and is inhibited by picrotoxin. Joint application of glutamate and GABA induces a nonadditive current under voltage clamp, suggesting that the transmitters can activate the same postsynaptic receptors. Immunocytochemical staining shows that, whereas some synaptic profiles impinging on the MoG contain pleomorphic agranular vesicles and are immunoreactive to GABA and not glutamate (as previously reported), there are at least as many other profiles that contain round, agranular vesicles and that are immunoreactive to glutamate and not to GABA. Thus, the authors conclude that some of the interneurones mediating inhibition of the electrical synapse use glutamate as their neurotransmitter.

Case-control study of hospital admission with asthma in children aged 5-14 years: relation with road traffic in north west London.

BACKGROUND: Evidence for an association between road traffic pollution and asthma is inconclusive. We report a case-control study of hospital admissions for asthma and respiratory illness among children aged 5-14 in relation to proxy markers of traffic related pollution. METHODS: The study was based on routine hospital admissions data in 1992/3 and 1993/4 for North Thames (West) health region within the M25 motorway. Cases were defined as emergency admissions for asthma (n = 1380) or all respiratory illness including asthma (n = 2131), and controls (n = 5703) were other emergency admissions excluding accidents. Cases and controls were compared with respect to distance of residence from nearest main road or roads with peak hour traffic >1000 vehicles and traffic volume within 150 m of residence, obtained by Geographical Information System techniques. Statistical analysis included adjustment for age, sex, admitting hospital, and a deprivation score for the census enumeration district of residence. RESULTS: Adjusted odds ratios of hospital admission for asthma and respiratory illness for children living within 150 m of a main road compared with those living further away were, respectively, 0.93 (95% CI 0.82 to 1.06) and 1.02 (95% CI 0.92 to 1.14). CONCLUSIONS: This study showed no association between risk of hospital admission for asthma or respiratory illness among children aged 5-14 and proxy markers of road traffic pollution.

Magnesium in drinking water supplies and mortality from acute myocardial infarction in north west England.

OBJECTIVES: To examine whether higher concentrations of magnesium in drinking water supplies are associated with lower mortality from acute myocardial infarction at a small area geographical level; to examine if the association is modified by age, sex, and socioeconomic deprivation. DESIGN: Small area geographical study using 13,794 census enumeration districts. Water constituent concentrations (magnesium, calcium, fluoride, lead) measured at water supply zone and assigned to enumeration districts. SETTING: 305 water supply zones in north west England. SUBJECTS: Resident population of 1,124,623 men and 1,372,036 women (1991 census) aged 45 years or more. MAIN OUTCOME MEASURE: Mortality from acute myocardial infarction, International Classification of Diseases, ninth revision (ICD-9) 410. Subsidiary analysis examined deaths from ischaemic heart disease, ICD 410-414. RESULTS: There were 21,339 male and 17,883 female deaths from acute myocardial infarction in 1990-92. Drinking water magnesium concentrations in water zones ranged from 2 mg/l to 111 mg/l (mean (SD) 19 (20) mg/l, median 12 mg/l); 24% of variation in magnesium concentrations was within zone and 76% was between zone. The relative risk of mortality from acute myocardial infarction (standardised for age, sex, and Carstairs deprivation quintile) for a quadrupling of magnesium concentrations in drinking water (for example, 20 mg/l v 5 mg/l) was 1.01 (95% confidence interval (CI) 0.99 to 1.03). When adjusted for north-south and east-west trends in mortality from acute myocardial infarction and for drinking water calcium, fluoride, and lead concentrations, this relative risk was 1.01 (95% CI 0.96 to 1.06). There was no evidence of a protective effect for acute myocardial infarction even among age, sex, and deprivation groups that were likely to be relatively magnesium deficient. For ischaemic heart disease mortality there was an apparent protective effect of magnesium and calcium (with calcium predominating in the joint model), but these were no longer significant when the geographical trends were incorporated. CONCLUSIONS: No evidence was found of an association between magnesium concentrations in drinking water supplies and mortality from acute myocardial infarction. These results do not support the hypothesis that magnesium is the key water factor in relation to mortality from heart disease.

Mapping the prevalence of sickle cell and beta thalassaemia in England: estimating and validating ethnic-specific rates.

A range of estimates for sickle cell and beta thalassaemia have been derived for the different ethnic groups living in the U.K., reflecting uncertainty over the true population value in certain countries and the heterogeneity within and between countries of origin comprising the same ethnic group. These were validated against six community screening programmes, with the estimated range correctly predicting the number of affected births observed by the programmes. In England approximately 3000 affected babies (0.47%) carry sickle cell trait and 2800 (0.44%) carry beta thalassaemia trait annually: with approximately 178 (0.28 per 1000 conceptions) affected by sickle cell disease (SCD) and 43 (0.07 per 1000) by beta thalassaemia major/intermedia. Allowing for termination, about 140-175 (0.22-0.28 per 1000) affected infants are born annually with SCD and from 10 to 25 (0.02-0.04 per 1000) with beta thalassaemia major/intermedia. These are the first evidence-based rates for sickle cell and beta thalassaemia for use in the U.K., and should underpin the future planning of services. The long-term solution to monitoring changes in the rates of trait and disease in the population is to introduce a standardized instrument for collecting ethnicity for all community screening programmes.

Lymphohaematopoietic malignancy around all industrial complexes that include major oil refineries in Great Britain.

OBJECTIVES: To examine the incidence of lymphohaematopoietic malignancy around industrial complexes that include major oil refineries in Great Britain after recent public and scientific concern of possible carcinogenic hazards of emissions from the petrochemical industry. METHODS: Small area study of the incidence of lymphohaematopoietic malignancies, 1974-91, within 7.5 km of all 11 oil refineries (grouped into seven sites) in Great Britain that were operational by the early 1970s and processed more than two million tonnes of crude oil in 1993. RESULTS: Combined analysis of data from all seven sites showed no significant (p < 0.05) increase in risk of these malignancies within 2 km or 7.5 km. Hodgkin's lymphoma, but no other malignancy, showed evidence (p = 0.02) of a decline in risk with distance from refineries, but there was an apparent deficit of cases of multiple myeloma near the refineries (p = 0.04). CONCLUSION: There was no evidence of association between residence near oil refineries and leukaemias, or non-Hodgkin's lymphoma. A weak positive association was found between risk of Hodgkin's disease and proximity to major petrochemical industry, and a negative association with multiple myeloma, which may be chance findings within the context of multiple statistical testing.

Actions of FMRFamide-related peptides on the gCa2+ of the C1 neuron in Helix aspersa.

The endogenous neuropeptides FMRFamide and FLRFamide (tetrapeptides) reversibly reduced a voltage-activated calcium current in the C1 neuron of Helix aspersa by an average of 20%. Two structurally related heptapeptides, pQDPFLRFamide and pQDPFLRIamide, both derived from another precursor protein in this species, did not reduce the current at all.

The peptide pQFYRFamide is encoded on the FMRFamide precursor of the snail Helix aspersa but does not activate the FMRFamide-gated sodium current.

The complete sequence of the FMRFamide precursor cDNA from Helix aspersa is reported here. Since the 5' end of this cDNA is identical to that of the precursor that encodes the heptapeptide analogs of FLRFamide, the two transcripts are probably derived by alternative RNA splicing. A novel pentapeptide, Glp-Phe-Tyr-Arg-Phe-NH2 (pQFYRFamide), predicted from the cDNA sequence, was purified from extracts of H. aspersa ganglia and identified by mass spectroscopy. Partial gene sequences for the FMRFamide precursors of two closely related pulmonate species-Cepaea nemoralis and Polydontes acutangula-were also determined and compared with the cDNA sequence of H. aspersa and a partial gene sequence previously determined from H. pomatia. Not only are the FMRFamide-related sequences and proteolytic processing sites conserved, but the linear arrangement of these landmarks is also retained. Synthetic pQFYRFamide has some effects on the isolated heart and on neuronal potassium currents in H. aspersa that are similar to those of FMRFamide, but it does not activate the neuronal FMRFamide-gated sodium channel.

Biliary nonmucin glycoproteins in patients with and without gallstones.

Total protein, mucin, and specific nonmucin glycoproteins are proposed pronucleating agents in cholesterol gallstone pathogenesis. However, characterization of specific nonmucin glycoproteins in patients with and without gallstones is unknown. Furthermore, nonmucin glycoproteins may be qualitatively different in patients with and without gallstones. Total protein and total and specific nonmucin glycoproteins were studied in gallbladder bile of 43 patients with cholesterol gallstones and 13 patients without gallstones. Patients with cholesterol gallstones had higher concentrations of both total protein and nonmucin glycoproteins than that observed in control patients (P < 0.05). SDS gel electrophoresis of nonmucin glycoproteins demonstrated an 84-kDa protein that was present significantly more often in patients with cholesterol gallstones (87% vs 8%, P < 0.05). Proposed 130- and 42-kDa pronucleating and 120-kDa anti-nucleating nonmucin glycoproteins were present in similar percentages in gallstone and control bile. Moreover, gallbladder bile of patients with the 84-kDa protein nucleated 50% faster than model bile and > 100% faster than that of patients without this protein (P < 0.05). The currently described gallbladder pronucleating and anti-nucleating proteins are found with equal frequency in cholesterol gallstone and control patients. However, an 84-kDa protein is found more commonly in gallstone patients and was associated with a shortened crystal observation time. Thus, this glycoprotein may be important in cholesterol gallstone pathogenesis.

The neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) directly gates two ion channels in an identified Helix neurone.

FMRFamide (i.e. Phe-Met-Arg-Phe-NH2) application to the C2 neurone of Helix caused a depolarizing response which consisted of a large, rapidly developing, and rapidly desensitizing inward current, underlain by a smaller, slower inward current which did not desensitize. Both currents were carried through sodium-selective channels which were insensitive to D-tubocurarine, and the to the fast sodium channel blockers tetrodotoxin (TTX) and lignocaine. Only the faster, desensitizing current could be blocked by amiloride. FMRFamide also activated two types of unitary inward currents with slightly differing amplitudes in outside-out patches taken from the C2 neurone, both through sodium-selective ion channels. Only the smaller unitary currents readily desensitized and were susceptible to block by amiloride, and they also activated more rapidly. Unitary currents of both types were recorded in outside-out patches in the absence of freely diffusible intracellular mediators, and were also activated when guanosine 5'-O-(2-thiodiphosphate) (GDP [beta-S]) was included in the recording pipette solution. This supports a tight receptor/channel coupling for both responses, with no involvement of GTP-binding proteins. Further, the very fast rate of activation of the smaller channels, which generally carry the major part of the FMRFamide-induced current, strongly indicates that these channels are ligand gated.