Genome-wide association study identifies a novel locus for cannabis dependence.

Despite moderate heritability, only one study has identified genome-wide significant loci for cannabis-related phenotypes. We conducted meta-analyses of genome-wide association study data on 2080 cannabis-dependent cases and 6435 cannabis-exposed controls of European descent. A cluster of correlated single-nucleotide polymorphisms (SNPs) in a novel region on chromosome 10 was genome-wide significant (lowest P=1.3E-8). Among the SNPs, rs1409568 showed enrichment for H3K4me1 and H3K427ac marks, suggesting its role as an enhancer in addiction-relevant brain regions, such as the dorsolateral prefrontal cortex and the angular and cingulate gyri. This SNP is also predicted to modify binding scores for several transcription factors. We found modest evidence for replication for rs1409568 in an independent cohort of African American (896 cases and 1591 controls; P=0.03) but not European American (EA; 781 cases and 1905 controls) participants. The combined meta-analysis (3757 cases and 9931 controls) indicated trend-level significance for rs1409568 (P=2.85E-7). No genome-wide significant loci emerged for cannabis dependence criterion count (n=8050). There was also evidence that the minor allele of rs1409568 was associated with a 2.1% increase in right hippocampal volume in an independent sample of 430 EA college students (fwe-P=0.008). The identification and characterization of genome-wide significant loci for cannabis dependence is among the first steps toward understanding the biological contributions to the etiology of this psychiatric disorder, which appears to be rising in some developed nations.

Evidence of CNIH3 involvement in opioid dependence.

Opioid dependence, a severe addictive disorder and major societal problem, has been demonstrated to be moderately heritable. We conducted a genome-wide association study in Comorbidity and Trauma Study data comparing opioid-dependent daily injectors (N=1167) with opioid misusers who never progressed to daily injection (N=161). The strongest associations, observed for CNIH3 single-nucleotide polymorphisms (SNPs), were confirmed in two independent samples, the Yale-Penn genetic studies of opioid, cocaine and alcohol dependence and the Study of Addiction: Genetics and Environment, which both contain non-dependent opioid misusers and opioid-dependent individuals. Meta-analyses found five genome-wide significant CNIH3 SNPs. The A allele of rs10799590, the most highly associated SNP, was robustly protective (P=4.30E-9; odds ratio 0.64 (95% confidence interval 0.55-0.74)). Epigenetic annotation predicts that this SNP is functional in fetal brain. Neuroimaging data from the Duke Neurogenetics Study (N=312) provide evidence of this SNP's in vivo functionality; rs10799590 A allele carriers displayed significantly greater right amygdala habituation to threat-related facial expressions, a phenotype associated with resilience to psychopathology. Computational genetic analyses of physical dependence on morphine across 23 mouse strains yielded significant correlations for haplotypes in CNIH3 and functionally related genes. These convergent findings support CNIH3 involvement in the pathophysiology of opioid dependence, complementing prior studies implicating the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate system.

Genome-wide association study on detailed profiles of smoking behavior and nicotine dependence in a twin sample.

Smoking is a major risk factor for several somatic diseases and is also emerging as a causal factor for neuropsychiatric disorders. Genome-wide association (GWA) and candidate gene studies for smoking behavior and nicotine dependence (ND) have disclosed too few predisposing variants to account for the high estimated heritability. Previous large-scale GWA studies have had very limited phenotypic definitions of relevance to smoking-related behavior, which has likely impeded the discovery of genetic effects. We performed GWA analyses on 1114 adult twins ascertained for ever smoking from the population-based Finnish Twin Cohort study. The availability of 17 smoking-related phenotypes allowed us to comprehensively portray the dimensions of smoking behavior, clustered into the domains of smoking initiation, amount smoked and ND. Our results highlight a locus on 16p12.3, with several single-nucleotide polymorphisms (SNPs) in the vicinity of CLEC19A showing association (P<1 × 10(-6)) with smoking quantity. Interestingly, CLEC19A is located close to a previously reported attention-deficit hyperactivity disorder (ADHD) linkage locus and an evident link between ADHD and smoking has been established. Intriguing preliminary association (P<1 × 10(-5)) was detected between DSM-IV (Diagnostic and Statistical Manual of Mental Disorders, 4th edition) ND diagnosis and several SNPs in ERBB4, coding for a Neuregulin receptor, on 2q33. The association between ERBB4 and DSM-IV ND diagnosis was replicated in an independent Australian sample. Recently, a significant increase in ErbB4 and Neuregulin 3 (Nrg3) expression was revealed following chronic nicotine exposure and withdrawal in mice and an association between NRG3 SNPs and smoking cessation success was detected in a clinical trial. ERBB4 has previously been associated with schizophrenia; further, it is located within an established schizophrenia linkage locus and within a linkage locus for a smoker phenotype identified in this sample. In conclusion, we disclose novel tentative evidence for the involvement of ERBB4 in ND, suggesting the involvement of the Neuregulin/ErbB signalling pathway in addictions and providing a plausible link between the high co-morbidity of schizophrenia and ND.

Cough sensors. I. Physiological and pharmacological properties of the afferent nerves regulating cough.

The afferent nerves regulating cough have been reasonably well defined. The selective effects of general anesthesia on C-fiber-dependent cough and the opposing effects of C-fiber subtypes in cough have led to some uncertainty about their regulation of this defensive reflex. But a role for C-fibers in cough seems almost certain, given the unique pharmacological properties of these unmyelinated vagal afferent nerves and the ability of many C-fiber-selective stimulants to evoke cough. The role of myelinated laryngeal, tracheal, and bronchial afferent nerve subtypes that can be activated by punctate mechanical stimuli, inhaled particulates, accumulated secretions, and acid has also been demonstrated. These "cough receptors" are distinct from the slowly and rapidly adapting intrapulmonary stretch receptors responding to lung inflation. Indeed, intrapulmonary rapidly and slowly adapting receptors and pulmonary C-fibers may play no role or a nonessential role in cough, or might even actively inhibit cough upon activation. A critical review of the studies of the afferent nerve subtypes most often implicated in cough is provided.

Effects of systemic administration of mirtazapine on respiratory muscle activity in sleeping rats.

Mirtazapine (MIRT) is an antidepressant with mixed noradrenergic and serotonergic effects in central nervous system. The present study was undertaken to assess whether MIRT can stimulate genioglossus muscle (GG) activity in the conscious, behaving rat. Nine male rats were chronically instrumented with GG and neck muscle EMG electrodes. EEG electrodes were implanted to acquire sleep stage. Results demonstrated a dose-dependent effect of MIRT on GG activity during sleep, although no changes reached statistical significance. Low dose MIRT (0.1 mg/kg) showed a slight increase in GG phasic activity. In contrast, higher doses of MIRT (0.5-1.0 mg/kg) tended to decrease GG activity relative to vehicle, in addition to decreasing total sleep time.

Genome-wide polygenic scores for age at onset of alcohol dependence and association with alcohol-related measures.

Age at onset of alcohol dependence (AO-AD) is a defining feature of multiple drinking typologies. AO-AD is heritable and likely shares genetic liability with other aspects of alcohol consumption. We examine whether polygenic variation in AO-AD, based on a genome-wide association study (GWAS), was associated with AO-AD and other aspects of alcohol consumption in two independent samples. Genetic risk scores (GRS) were created based on AO-AD GWAS results from a discovery sample of 1788 regular drinkers from extended pedigrees from the Collaborative Study of the Genetics of Alcoholism (COGA). GRS were used to predict AO-AD, AD and Alcohol dependence symptom count (AD-SX), age at onset of intoxication (AO-I), as well as maxdrinks in regular drinking participants from two independent samples-the Study of Addictions: Genes and Environment (SAGE; n=2336) and an Australian sample (OZ-ALC; n=5816). GRS for AO-AD from COGA explained a modest but significant proportion of the variance in all alcohol-related phenotypes in SAGE. Despite including effect sizes associated with large numbers of single nucleotide polymorphisms (SNPs; >110 000), GRS explained, at most, 0.7% of the variance in these alcohol measures in this independent sample. In OZ-ALC, significant but even more modest associations were noted with variance estimates ranging from 0.03 to 0.16%. In conclusion, there is modest evidence that genetic variation in AO-AD is associated with liability to other aspects of alcohol involvement.

Movement of finger joints induced by synergistic wrist motion.

BACKGROUND: An experiment has recently been conducted to evaluate and compare the differences in tendon excursions between the flexor digitorum profundus and superficialis using three mobilization techniques. No previous studies deal with the total joint excursions with constant tendon length. The purpose of this study was to investigate the coordinated motion between the finger and wrist joints resulting from passive tension of the muscles while performing synergistic wrist motion. METHODS: The relative joint positions of the hand and wrist were measured using a three-dimensional motion analysis system with external retroreflective markers 2 mm in diameter placed on the dorsal surface of the hand. Fifty normal subjects, with a 1:1 gender ration, ranging in age from 20 to 40 years, and with no previous history of upper extremity injury, were recruited for the experiment. FINDINGS: The relationships of synergistic motion between the wrist and finger joints due to passive tension in the muscles were approximately linear. The ranges of wrist motion averaged 60 degrees extension and 60 degrees flexion. Moving the wrist from flexion into extension induced synergistic finger joint motion as follows: the distal interphalangeal joint angles changed from an average of 12 degrees of flexion to 31 degrees; proximal-interphalangeal joint angles changed from 19 degrees to 70 degrees; and metacarpal phalangeal joints changed from 27 degrees to 63 degrees of flexion. INTERPRETATION: The relationships of synergistic motion between the wrist and finger joints were systematically documented. Such a relationship could be considered in optimizing the design of dynamic splints used for rehabilitation in post-surgical tendon repair, as well as providing useful information about potential diagnoses of problems with the integrity of the flexor and extensor mechanisms.

Synthesis and biological activity of angiotensin II analogues containing a Val-His replacement, Val psi[CH(CONH2)NH]His.

The dipeptide mimic Val psi[CH(CONH2)NH]His (4) was incorporated into angiotensin II (AII) analogues to provide an octapeptide saralasin derivative (29) as well as tetrapeptide analogue 19. Three C-terminal tetrapeptides (21, 25, and 28) were also prepared. All compounds were tested for their ability to displace 3H-AII from rabbit adrenal gland homogenate and as antagonists of AII and AI on guinea pig ileum. The octapeptide analogue 29 was 700 times less active than the parent peptide 30. All the C-terminal fragments 19, 21, 25, and 28 have no measurable AII antagonist activity. Of the four tetrapeptide fragments, only 21 showed any appreciable binding activity.

(Imidazolylphenyl)pyrrol-2-one inhibitors of cardiac cAMP phosphodiesterase.

Seven 3-alkyl-4-aryl-1,5-dihydro-2H-pyrrol-2-ones were prepared as potential inhibitors of cardiac cAMP phosphodiesterase (PDE). The design of these compounds made use of rolipram, a known inhibitor of the brain cAMP PDE isozyme, as a lead structure and was guided by a model which describes the features required for potent inhibition of the cardiac isozyme. Syntheses for the new compounds are described, together with the results of theoretical and crystallographic studies aimed toward ascertaining their three-dimensional structures. The activities of these compounds as inhibitors of the cardiac and brain cAMP PDE isozymes and their positive inotropic activity in ferret papillary muscle are also reported. Selected compounds were further examined in an in vivo hemodynamic model. One compound 1,5-dihydro-4-[4-(1H-imidazol-1- yl)phenyl]-3-methyl-2H-pyrrol-2-one, was identified as a potent and selective positive inotropic agent and inhibitor of cardiac cAMP PDE.

Cardiotonic agents. 2. (Imidazolyl)aroylimidazolones, highly potent and selective positive inotropic agents.

A series of 4-alkyl-1,3-dihydro-5-[(1H-imidazolyl)benzoyl]-2H-imidazol-2-ones 9 was synthesized and evaluated in vitro for positive inotropic and cyclic AMP phosphodiesterase inhibitory activity. A wide range of inotropic and enzyme-inhibitory potencies was observed, substitution on the imidazolyl moiety being the major determinant of activity. The 4-ethyl-5-[4-(1H-imidazol-1-yl)benzoyl] congener 9g exhibited the highest potency in vitro. Incorporation of a methyl group at the imidazolyl 2-position gave 9h, which was less potent but remarkably selective in vivo for positive inotropic effects over heart rate and hypotensive effects.

Cardiotonic agents. 6. Histamine analogues as potential cardiovascular selective H2 agonists.

Twenty-six alkyl and aralkyl histamine analogues were prepared as potential cardiotonic agents. Compounds were designed to allow interaction with a putative secondary aryl binding site at the H2 receptor, the presence of which was inferred from the structure of cyprohepatadine, which is known to have H2-antagonist properties. The compounds were examined for inotropic activity in ferret papillary muscle. Potent inotropic activity was generally found in N-alkyl- and N,N-dialkylimidazole-4-ethanamines, whereas N-(amidoalkyl)imidazole-4-ethanamines and N-alkylimidazole-4-propanamines were at best weakly active. Five compounds were examined in screens designed to assess hemodynamic effects and gastric acid secretion in vivo. Two of these compounds, alpha-(3-phenyl-2-transpropenyl)-1H-imidazole-4-ethanamine and N-heptyl-1H-imidazole-4-ethanamine, showed positive inotropic activity with minimal effects on heart rate and mean arterial pressure in vivo; however, both compounds were found to stimulate gastric acid secretion. These results demonstrate that selectivity between various H2-receptor-mediated activities can be obtained with substituted histamine analogues.

A topographical model for the c-AMP phosphodiesterase III active site.

With this minireview, concepts about how c-AMP and various inhibitor molecules interact with the phosphodiesterases seem to have come full-circle. It will be proposed and elaborated herein that an understanding of SAR for the newest, "second generation" PDE inhibitors is best accomplished by adopting a model that supposes that these compounds are transition state inhibitors. The analysis finds an interesting parallel with early studies where it was recognized that c-AMP adopts a trigonal bipyramid transition state during hydrolysis. The dynamic interaction of ligands with the phosphodiesterase enzymes will also be made evident when simple algebraic expressions are shown to be inadequate for predicting inhibitor potencies. The latter are apparently complicated by cooperative or synergistic relationships that occur among the various binding sites within the receptor. Finally, implications that can be derived from certain topographical features of the model are discussed relative to a range of potential therapeutic indications.

Gait analysis and energy consumption of below-knee amputees wearing three different prosthetic feet.

This study scientifically measures the dynamic gait characteristics and energy consumption of 16 male below-knee amputees, eight vascular and eight traumatic, while wearing solid ankle cushion heel (SACH), single axis and multiple axis prosthetic feet via six-camera motion analysis, metabolic measurement cart and heavy-duty treadmill. Subjective results are additionally determined via questionnaire after testing. Motion analysis showed statistically significant differences at P<0.05 between the SACH, single axis and multiple axis foot in the velocity, cadence, stride length and single limb stance. Significant differences were found in energy consumption between the traumatic and vascular groups, and significant changes in walking under different speeds and different inclines. Results provide quantitative and qualitative information about the dynamic performance of the various feet, which can be helpful in prescribing the optimal prosthetic foot for individual amputees.

Common abnormal kinetic patterns of the knee in gait in spastic diplegia of cerebral palsy.

We studied the kinetic characteristics of the knee in patients with spastic diplegia. Twenty three children with spastic diplegia were recruited and had their 46 limbs categorised into the following four groups: jump (n=7), crouch (n=8), recurvatum (n=14) and mild (n=17). In the crouch pattern, the patients usually had a larger and longer lasting internal knee extensor moments in stance suggesting that rectus femoris had a relatively high activation. In the recurvatum pattern, the internal knee flexor moment was large and long lasting in stance. The biceps femoris showed less activity on EMG although the knee flexor moment was large and we concluded that the soft tissue behind the knee joint provided this flexor moment. In the jump knee pattern there was abnormal power generation at the knee and ankle joints in initial stance, which did not contribute to normal progression but aided upward body motion. In the mild group the kinetic data was similar to that seen in normal children. Knowledge of kinetic patterns in these patients may help in their subsequent management.

The effect of changing the foot progression angle on the knee adduction moment in normal teenagers.

The effect of changing the foot progression angle on the peak knee adduction moment (KAM) during stance was investigated in 48 teenagers. They underwent gait analysis when walking in three different postures: normal walking, intentional in-toeing, and intentional out-toeing. The peak KAM when in-toeing was the highest and was statistically different from that seen in the normal walking or in the out-toeing posture. These findings may have clinical significance in adult life.

Gait analysis after ankle arthrodesis.

The purpose of this study was to employ a computerized motion analysis system to identify the effect of ankle arthrodesis on the three-dimensional kinematic behavior of the rear and fore foot during level walking. A three-segment rigid body model was used to describe the motion of the foot and ankle. The results demonstrated that sagittal plane motion of the hindfoot was significantly decreased in the foot of patients having had ankle arthrodesis compared to normal subjects. The kinematic data indicated a generalized stiffness of the hindfoot on the involved foot in the sagittal plane. Sagittal plane movement in the forefoot and transverse plane movements in the hindfoot and forefoot increased in patients compared to controls.

Measurement of organ volume by single photon emission computed tomography.

The aim of this study was to develop a new algorithm for volume determination by single photon emission computed tomography (SPECT). Different algorithms were evaluated through phantom studies. The results show that the algorithm combining moment-preserving bilevel thresholding and best-fit Laplacian second derivative edge detection can provide the most accurate measurement of volume. Besides, this method can be utilized in different SPECT systems with no need for further phantom studies. In patient studies, the results of liver volume calculation have indicated that this is a useful technique in clinical medicine.

Evaluation of enzyme-linked immunosorbent assay for the determination of polycyclic aromatic hydrocarbons in house dust and residential soil.

Two commercially available enzyme-linked immunosorbent assays (ELISA) for total polycyclic aromatic hydrocarbon (PAH) and carcinogenic PAH (C-PAH) were evaluated. The testing procedures were refined for application to screening PAH and C-PAH in house dust and soil samples for human exposure studies. The overall method precision expressed as percent relative standard deviation (%RSD) of triplicate real world dust and soil samples was within +/- 29% (12-29%) for PAH ELISA and +/- 21% (5.9-21%) for C-PAH ELISA. Spike recoveries from real world dust/soil samples were 114 +/- 30% for phenanthrene from PAH ELISA and 120 +/- 8.2% for benzo[a]pyrene from C-PAH ELISA. The overall method accuracy for PAH and C-PAH assays cannot be assessed for multiple PAH components in dust/soil samples (which represent real-world samples), because of the assays' cross reactivities with other PAH components. Over 100 dust/soil samples from 13 North Carolina homes and 22 Arizona homes were analyzed by PAH and C-PAH assays, as well as by the conventional gas chromatography/mass spectrometry (GC/MS) method. Statistical analysis showed that dust/soil PAH data from ELISA and GC/MS methods are significantly different. In general PAH ELISA responses were higher than PAH GC/MS responses. The regression analysis showed that the linear relationship between ELISA and GC/MS measurements is not strong in the combined data. The relationship became stronger for the data from the same type of dust/soil samples. The screening performance of ELISA was evaluated based on the frequency distribution of ELISA and GC/MS data. The results indicated that the ELISA PAH and C-PAH assays cannot be used as a quantitative analytical tool for determining PAH in real-world dust/soil samples. However, the ELISA is an effective screening tool for ranking PAH concentrations in similar types of real world dust/soil samples.

Foot progression angle and ankle joint complex in preschool children.

OBJECTIVE: The influence of foot progression angle on the ankle joint and the effects on gait patterns and mechanisms in skeletally normal preschool children was investigated. DESIGN: Kinematics and kinetics of the ankle joint were analyzed for preschool children who were skeletally normal but walked with different foot progression angle. BACKGROUND: The most frequent reasons for preschool children to be brought to a paediatric orthopaedic clinic are toe-in and toe-out. Without understanding the biological and biomechanical implications, treatment for these problems can be very confusing. METHODS: Gait analysis was performed in 86 skeletally normal preschool children. Children were grouped according to their foot progression angles. Analysis of the kinematics and kinetics of the ankle joint was intended to elucidate the gait mechanism. RESULTS: Children with different foot progression angles had distinctive patterns of spatio-temporal parameters, ground reaction force, joint angle, moment, power, and mechanical work of the ankle joints. The differences were organized and explained as different walking patterns and tactics. CONCLUSIONS: Skeletally normal preschool children with excessive toe-in or toe-out foot progression angles are not necessarily in some diseased status. They were instead related to different walking patterns. Aggressive treatment for these problems is not recommended.RelevanceThese results suggest that toe-in and toe-out are related to walking speed which has distinct influences on the kinematics and kinetics of the ankle joint. Though the observed problems were seemingly only in the transverse plane, they are in fact three-dimensional and have a mutually close relationship. The findings could be linked to the promptness of muscle response and the habits of walking in preschool children. Better understanding of possible mechanisms will help parents and paediatricians decide on the most appropriate treatment for these children.

The effect of magnitudes and duration of pressure on cerebral cortex in a rat model.

The aim of this biomechanical study was to investigate the pressure-time-damage relationship on the cerebral cortex using a rat model. During brain surgery, additional cortical injuries induced by traction have been an important clinical issue. A total of 84 rats underwent a unilateral craniectomy, and conduction by hydrostatic pressure loading through a modified central venous pressure device with various combinations of magnitudes ranging from 0.5 to 10 cm H2O and duration from 0.5 to 20 minutes was then performed. Histopathological examination has exhibited three patterns of clustered, spotted damaged neurons and undamaged neuron. Two best-fitted exponential curves were derived from the data to establish the damaged, critical and tolerable loadings responsible for the neuron viability. This research provides information to enhance understanding of the additional cortex injuries induced by traction. Furthermore, the results may have implications in providing clinical guidance and/or development of alarm systems for minimising cortical damage during surgery.