CC16 polymorphisms in asthma, asthma subtypes, and asthma control in adults from the Agricultural Lung Health Study.

BACKGROUND: The club cell secretory protein (CC16) has anti-inflammatory and antioxidant effects and is a potential early biomarker of lung damage. The CC16 single nucleotide polymorphism (SNP) rs3741240 risk allele (A) has been inconsistently linked to asthma; other tagging SNPs in the gene have not been explored. The aim was to determine whether CC16 tagging polymorphisms are associated with adult asthma, asthma subtypes or asthma control in the Agricultural Lung Health Study (ALHS). METHODS: The ALHS is an asthma case-control study nested in the Agricultural Health Study cohort. Asthma cases were individuals with current doctor diagnosed asthma, likely undiagnosed asthma, or asthma-COPD overlap defined by questionnaire. We also examined asthma subtypes and asthma control. Five CC16 tagging SNPs were imputed to 1000 Genomes Integrated phase 1 reference panel. Logistic regression was used to estimate associations between CC16 SNPs and asthma outcomes adjusted for covariates. RESULTS: The sample included 1120 asthma cases and 1926 controls of European ancestry, with a mean age of 63 years. The frequency of the risk genotype (AA) for rs3741240 was 12.5% (n = 382). CC16 rs3741240 was not associated with adult asthma outcomes. A tagging SNP in the CC16 gene, rs12270961 was associated with uncontrolled asthma (n = 208, ORadj= 1.4, 95% CI 1.0, 1.9; p = 0.03). CONCLUSION: This study, the largest study to investigate associations between CC16 tagging SNPs and asthma phenotypes in adults, did not confirm an association of rs3741240 with adult asthma. A tagging SNP in CC16 suggests a potential relationship with asthma control.

Asymmetric and Distant Effects of a Unilateral Lesion of the Primary Motor Cortex on the Bilateral Supplementary Motor Areas in Adult Macaque Monkeys.

A restricted lesion of the hand area in the primary motor cortex (M1) leads to a deficit of contralesional manual dexterity, followed by an incomplete functional recovery, accompanied by plastic changes in M1 itself and in other cortical areas on both hemispheres. Using the marker SMI-32 specific to pyramidal neurons in cortical layers III and V, we investigated the impact of a focal unilateral M1 lesion (hand representation) on the rostral part (F6) and caudal part (F3) of the supplementary motor area (SMA) in both hemispheres in nine adult macaque monkeys compared with four intact control monkeys. The M1 lesion induced a consistent interhemispheric asymmetry in density of SMI-32-positive neurons in F3 layer V (statistically significant in 8 of 9 lesioned monkeys), highly correlated with the lesion volume and with the duration of functional recovery, but not with the extent of functional recovery itself. Such interhemispheric asymmetry was neither present in the intact monkeys, as expected, nor in F6 in all monkeys. In addition, the M1 lesion also impacted on the basal dendritic arborization of F3 layer V neurons. Neuronal density was clearly less affected by the M1 lesion in F3 layer III compared with layer V. We interpret the remote effect of M1 lesion onto the density of SMI-32-positive neurons and dendritic arborization in the SMAs bilaterally as the consequence of multiple factors, such as changes of connectivity, diaschisis and various mechanisms involved in cortical plasticity underlying the functional recovery from the M1 lesion.SIGNIFICANCE STATEMENT The motor system of macaque monkeys, in addition to be similarly organized as in humans, is a good candidate to study the impact of a focal lesion of the main contributor to voluntary movements, the primary motor cortex (M1), on non-primary motor cortical areas also involved in manual dexterity, both at behavioral and structural levels. Our results show that a unilateral permanent lesion of M1 hand area in nine monkeys affects the interhemispheric balance of the number of SMI-32-positive pyramidal neurons in the cortical layer V of the supplementary motor area, in a way strongly correlated to the lesion volume and duration of the incomplete functional recovery.

Prevalence of human papillomavirus among oesophageal squamous cell carcinoma cases: systematic review and meta-analysis.

BACKGROUND: Oncogenic human papillomavirus (HPV) has been hypothesised as a risk factor for oesophageal squamous cell carcinoma (OSCC), but aetiological research has been limited by the varying methodology used for establishing HPV prevalence. The aims of this systematic review and meta-analysis were to estimate the prevalence of HPV DNA detected in OSCC tumours and the influence of study characteristics. METHODS: Study-level estimates of overall and type-specific HPV prevalence were meta-analysed to obtain random-effects summary estimates. RESULTS: This analysis included 124 studies with a total of 13 832 OSCC cases. The average HPV prevalence (95% confidence interval) among OSCC cases was 0.277 (0.234, 0.320) by polymerase chain reaction; 0.243 (0.159, 0.326) by in situ hybridisation; 0.304 (0.185, 0.423) by immunohistochemistry; 0.322 (0.154, 0.490) by L1 serology; and 0.176 (0.061, 0.292) by Southern/slot/dot blot. The highest HPV prevalence was found in Africa and Asia, notably among Chinese studies from provinces with high OSCC incidence rates. CONCLUSIONS: Future research should focus on quantifying HPV in OSCC cases using strict quality control measures, as well as determining the association between HPV and OSCC incidence by conducting large, population-based case-control studies. Such studies will provide a richer understanding of the role of HPV in OSCC aetiology.

Prevalence, location, grade of pressure ulcers and association with specific patient characteristics in adult spinal cord injury patients during the hospital stay: a prospective cohort study.

STUDY DESIGN: This was a prospective cohort study. OBJECTIVES: The objective was to describe the incidence, prevalence, characteristics of pressure ulcers (PUs) and the association with specific patient characteristics in a consecutive sample of in-patients with a spinal cord injury (SCI). SETTING: An acute care and rehabilitation clinic specialized in SCIs in Switzerland. METHODS: The presence and characteristics of PUs for all adult patients with a SCI admitted to the clinic from 1 September 2009 to 28 February 2010 were recorded on a daily basis during their complete hospitalization. Risk factors were analyzed in univariate and multivariate logistic regression models. RESULTS: A total of 185 patients were included in the study and observed for the entirety of their hospitalization. The prevalence of at least one PU was 49.2% in all patients, compared with 25.4% in the group of patients admitted without PUs. The incidence was 2.2 per person and year. In 91 patients, a total of 219 PUs were observed. PUs were most frequently located on the foot (36.1%), and the coccyx/sacrum (15.1%). The risk for occurrence of a PU increased with age (odds ratio (OR)=1.04) and post SCI (OR=1.03). In the multivariate analyses, the risk for PUs was lower for patients with the American Spinal Injury Association (ASIA) Impairment Scale (AIS) of C or D (ORC=0.25, ORD=0.28) compared with patients with an AIS of A. CONCLUSION: Using a daily documentation system, PUs were detected as a frequent complication of SCIs. Completeness of injury, age and time since injury were significant risk factors for PUs. The foot was a region at high risk for PUs.

Development and validation of a self-report version of the Spinal Cord Independence Measure (SCIM III).

STUDY DESIGN: Cross-sectional validation study. OBJECTIVES: To develop and validate a self-report version of the Spinal Cord Independence Measure (SCIM III). SETTING: Two SCI rehabilitation facilities in Switzerland. METHODS: SCIM III comprises 19 questions on daily tasks with a total score between 0 and 100 and subscales for 'self-care', 'respiration & sphincter management' and 'mobility'. A self-report version (SCIM-SR) was developed by expert discussions and pretests in individuals with spinal cord injury (SCI) using a German translation. A convenience sample of 99 inpatients with SCI was recruited. SCIM-SR data were analyzed together with SCIM III data obtained from attending health professionals. RESULTS: High correlations between SCIM III and SCIM-SR were observed. Pearson's r for the total score was 0.87 (95% confidence interval (CI) 0.82-0.91), for the subscales self-care 0.87 (0.81-0.91); respiration & sphincter management 0.81 (0.73-0.87); and mobility 0.87 (0.82-0.91). Intraclass correlations were: total score 0.90 (95% CI 0.85-0.93); self-care 0.86 (0.79-0.90); respiration & sphincter management 0.80 (0.71-0.86); and mobility 0.83 (0.76-0.89). Bland-Altman plots showed that patients rated their functioning higher than professionals, in particular for mobility. The mean difference between SCIM-SR and SCIM III for the total score was 5.14 (point estimate 95% CI 2.95-7.34), self-care 0.89 (0.19-1.59), respiration & sphincter management 1.05 (0.18-2.28 ) and mobility 3.49 (2.44-4.54). Particularly patients readmitted because of pressure sores rated their independence higher than attending professionals. CONCLUSION: Our results support the criterion validity of SCIM-SR. The self-report version may facilitate long-term evaluations of independence in persons with SCI in their home situation.

Skeleton of the Oldest Known Pinniped, Enaliarctos mealsi.

A nearly complete skeleton of the archaic pinniped Enaliarctos, found in late Oligocene or early Miocene rocks (approximately 23 million years old) of California, provides new evidence on the origin of pinnipeds. Enaliarctos retains many primitive features expected in the hypothesized common ancestor of pinnipeds. Skeletal modifications seen in Enaliarctos document swimming adaptations and indicate that pinnipeds primitively used the axial skeleton and both fore and hindflippers as sources of propulsion. Elongate hindlimbs with prominent bony processes (reflecting powerful musculature) suggest that Enaliarctos was more active on land than modern pinnipeds.

The Oxysterol Synthesising Enzyme CH25H Contributes to the Development of Intestinal Fibrosis.

Intestinal fibrosis and stenosis are common complications of Crohn's disease [CD], frequently requiring surgery. Anti-inflammatory strategies can only partially prevent fibrosis; hence, anti-fibrotic therapies remain an unmet clinical need. Oxysterols are oxidised cholesterol derivatives with important roles in various biological processes. The enzyme cholesterol 25-hydroxylase [CH25H] converts cholesterol to 25-hydroxycholesterol [25-HC], which modulates immune responses and oxidative stress. In human intestinal samples from CD patients, we found a strong correlation of CH25H mRNA expression with the expression of fibrosis markers. We demonstrate reduced intestinal fibrosis in mice deficient for the CH25H enzyme, using the sodium dextran sulphate [DSS]-induced chronic colitis model. Additionally, using a heterotopic transplantation model of intestinal fibrosis, we demonstrate reduced collagen deposition and lower concentrations of hydroxyproline in CH25H knockouts. In the heterotopic transplant model, CH25H was expressed in fibroblasts. Taken together, our findings indicate an involvement of oxysterol synthesis in the pathogenesis of intestinal fibrosis.

Myristylation and amino-terminal phosphorylation are required for activation of pp60c-src during mitosis.

pp60c-src, a cellular tyrosine kinase homologous to the retroviral v-src oncogene, becomes transiently activated during mitosis. Activation is accompanied by phosphorylation of three sites in the amino-terminal regulatory domain of the protein, threonine 34, threonine 46 and serine 72. These sites can be phosphorylated in vitro by a cell cycle-regulated kinase, p34cdc2, yet this does not result in increased kinase activity of pp60c-src. pp60c-src is negatively regulated by phosphorylation at tyrosine 527, and it has been shown that this site is transiently dephosphorylated in mitotic cells. The importance of tyrosine 527 in the regulation of pp60c-src is also emphasized by the fact that oncogenic mutants of pp60src lacking tyrosine 527 are constitutively active during the entire cell cycle. Here we report that a non-myristylated mutant of pp60c-src is not activated and only partially phosphorylated at the amino terminus in mitotic cells. Additional mutants lacking one (TTAc-src), two (AASc-src) and three (AAAc-src) cdc2 phosphorylation sites had slightly higher kinase activity than wild-type pp60c-src in interphase cells and were not activated during mitosis. However, all four mutant proteins were still transiently dephosphorylated at tyrosine 527 during mitosis, suggesting that myristylation and amino-terminal phosphorylation may be necessary but are clearly not sufficient for mitosis-specific activation.

Regulation of cytochrome P450 in the central nervous system.

The role of brain P450 in the physiology, pharmacology and toxicology of the brain is the subject of this study. Cytochrome P450 was isolated from the brains of rats and quantitated spectrally. The contribution of the known hepatic forms of the enzyme to the forms constitutive in the brain as well as those which are induced by hormones are xenobiotics were characterized on Western blots. We have found that the level of P450 in the brain is increased during pregnancy and lactation, by partial hepatectomy and by ethanol. In each case the profile of P450s induced is different. In pregnancy and lactation the P450 content of the hypothalamic preoptic area and olfactory lobes were increased up to 10-fold and the only subfamily identified on Western blots was 4A. There was no detectable 1A, 2A, 2B, 2C, or 2E1. Ethanol increases the level of brain P450 3- to 5-fold and P450 2C, 2E1 and 4A are induced. Upon partial hepatectomy P450 1A, 2C and 4A were detected on Western blots but there was no 2E1. The inducibility of these forms of P450 in the brain suggests that there is in situ metabolism of steroids, fatty acids, prostaglandins, ethanol and other xenobiotics in the brain and raises questions about the role of brain P450 in the development of tolerance to drugs and the neurotoxicity of xenobiotics. More importantly, the action of neurotransmitters such as dopamine which utilize fatty acids metabolites as intracellular mediators, could be influenced by the levels of 2C and 4A P450s.

A novel reactive perstraction system based on liquid-core microcapsules applied to lipase-catalyzed biotransformations.

A novel reactive perstraction system has been developed based on liquid-core capsules, involving an enzyme-catalyzed reaction coupled with simultaneous in situ product recovery. Lipase-catalyzed reactions, hydrolysis of triprionin and nitrophenyl laurate, were selected to test the system and demonstrate the feasibility of immobilization of enzymes to the membranes of liquid-core capsules and the ability to extract hydrophobic products of the reaction within the capsule core. The lipase from Candida rugosa was immobilized to the microcapsules by adsorption and by covalent binding through activation with glutaraldehyde. In both cases improved temperature and operational stability were achieved. Both types of immobilization resulted in a basic shift of the pH optimum for activity, from 7.5 to 9.0. The presence of an organic phase within the capsule core allowed direct product separation and lead to a decrease in product inhibition of the lipase-catalyzed reaction.

Novel reactive perstraction system applied to the hydrolysis of penicillin G.

The activity of penicillin acylase has been studied in aqueous and organic solvents, as free enzyme as well as immobilized within the membrane of liquid-core capsules. The activity of the enzyme is inhibited by the accumulation of the products of the hydrolysis reaction, namely phenyl acetic acid (PAA). In order to overcome this inhibition a range of organic solvents were tested for use in in situ product recovery. Of these solvents dibutyl sebacate (DBS) was chosen due to the rapid extraction rate, the high logP and to facilitate capsule production. The extraction efficiency at pH 3.5 for PAA was >80% for phase ratios of >50% free solvent with partition coefficients of 8 and 0.7 for PAA and penicillin G (PenG), respectively, thereby showing that PAA could be selectively extracted at pH 3.5 and 25 degrees C. Liquid-core capsules containing DBS were shown to efficiently remove PAA selectively and the PAA could be effectively back-extracted and the capsules re-used in a three-stage process resulting in high product separation. Immobilization of penicillin acylase onto the capsule membranes resulted in increased operational stability of the enzyme and a very high enzyme activity. Over 53.3% of the PAA formed could be recovered in the capsule core with a concentration over sevenfold higher than in the aqueous phase. Higher extraction efficiencies could be obtained by varying the substrate concentration and number of capsules. The enzyme immobilized on capsules could be stored for over 4 months at pH 8 and 4 degrees C with no loss of activity. Over 80% of the initial activity could be recovered over five repeated batch cycles of the bioconversion process. The importance of capsular perstraction and reactive capsular perstraction has been clearly demonstrated.

[Several years' experience with thumb saddle joint prosthesis by the de la Caffinière method]. / Mehrjährige Erfahrungen mit der Daumen-sattelgelenksprothese nach de la CAFFINIERE.

This type of endoprosthesis was used for four years in chronically painful idiopathic osteoarthritis of the carpometacarpal joint of the thumb. Our results match exactly those published by the originator of the endoprosthesis CAFFINIERE, i.e. 70% excellent results, nearly 30% problem cases. The problem of loosening of the stem or the cup of the prosthesis seems to depend in our cases on the operative technique rather than on the design or other factors. In the rheumatoid patient as well as in polyarthrotic hands we cannot encourage the prosthetic replacement of the carpometacarpal joint of the thumb.

Calcium-dependent and oncogenic IL-3 mRNA stabilization can be distinguished pharmacologically and by sequence requirements in the 3'UTR.

In interleukin-3 (IL-3)-dependent PB-3c mast cells, the normally short-lived IL-3 mRNA is stabilized upon calcium-ionophore treatment or following v-H-ras induced oncogenesis. We compared the underlying stabilization mechanisms by analysing the response to the post-transcriptionally acting drugs cyclosporin A (CsA), FK506 and SB202190. Stable IL-3 transcripts in the PB-3c-derived tumour cell line V2D1 decayed in response to CsA and FK506, but not in response to SB202190. Transcripts stabilized by elevating intracellular calcium levels in PB-3c cells were destabilized in response to all three drugs. In PB-3c cells, six AUUUA pentamers within the AU-rich element were sufficient to confer responsiveness to calcium-ionophore and CsA treatment. In V2D1 tumour cells, sensitivity to CsA required additional nucleotides flanking these pentamers. Our data suggest that IL-3 mRNA stabilization by either calcium-dependent or oncogenic pathways involves different intracellular mechanisms.

Recent advances in South American mammalian paleontology.

Recently discovered deposits containing terrestrial mammal fossils, together with multidisciplinary studies of classical sequences, have yielded dramatic insights into the biotic and environmental history of South America. Notable advances include several new fossil primate taxa, an improved chronology of two major immigration events (caviomorph rodents and new world monkeys), documentation of the oldest mammalian faunas dominated by grazing taxa (which suggests that grasslands appeared at least 15 million years earlier than on other continents), evidence of early biogeographical provinciality within South America, and improved sampling of the best known Cenozoic tropical South American paleofauna.

Molecular analysis of vitamin A formation: cloning and characterization of beta-carotene 15,15'-dioxygenases.

Beta-carotene 15,15'-dioxygenase cleaves beta-carotene into two molecules of retinal and is the key enzyme in the metabolism of carotene to vitamin A. Although the enzyme has been known for more than 40 years, all attempts to purify the protein to homogeneity or to clone its gene have failed until recently, when the successful cloning and sequencing of cDNAs encoding enzymes with beta-carotene 15,15'-dioxygenase activity from Drosophila (J. von Lintig and K. Vogt, 2000, J. Biol. Chem. 275, 11915-11920) and chicken (A. Wyss et al., 2000, Biochem. Biophys. Res. Commun. 271, 334-336) were reported. Very soon it became clear, that we have cloned two members of a new family of carotenoid cleaving enzymes. Overall homologies are very high, certain amino acid stretches almost identical. Thus, beta-carotene 15,15'-dioxygenase can be considered as evolutionarily well conserved. These findings open up wide perspectives for further analysis of this important biosynthetic pathway, concerning basic and medical research as well as biotechnological aspects related to vitamin A supply, which are discussed here.

Multituberculate and other mammal hair recovered from Palaeogene excreta.

Evidence of hair from several extinct mammals has been recovered from a rich accumulation of fossil excrement from the Late Palaeocene beds of Inner Mongolia, China. This highly unusual and previously undocumented depositional occurrence consists of hundreds of mammalian carnivore coprolites (fossil faeces) and a lesser number of probably raptorial bird regurgitalites (fossil pellets). The fossil hair occurs as impressions and natural casts in the extremely fine-grained, calcareous matrix that cements the skeletal remains within these faecal structures and preserves even the cuticular scale pattern on individual hair. Hair from at least four mammalian taxa, most notably the multituberculate Lambdopsalis bulla, has been identified. This record constitutes the first tangible evidence that, along with monotremes and therian mammals, multituberculates were hirsuite, and lends support for the presence of this mammalian feature in the most recent common ancestor of these three groups.