Solution structure of recombinant somatomedin B domain from vitronectin produced in Pichia pastoris.
The cysteine-rich somatomedin B domain (SMB) of the matrix protein vitronectin is involved in several important biological processes. First, it stabilizes the active conformation of the plasminogen activator inhibitor (PAI-1); second, it provides the recognition motif for cell adhesion via the cognate integrins (alpha(v)beta(3), alpha(v)beta(5), and alpha(IIb)beta(3)); and third, it binds the complex between urokinase-type plasminogen activator (uPA) and its glycolipid-anchored receptor (uPAR). Previous structural studies on SMB have used recombinant protein expressed in Escherichia coli or SMB released from plasma-derived vitronectin by CNBr cleavage. However, different disulfide patterns and three-dimensional structures for SMB were reported. In the present study, we have expressed recombinant human SMB by two different eukaryotic expression systems, Pichia pastoris and Drosophila melanogaster S2-cells, both yielding structurally and functionally homogeneous protein preparations. Importantly, the entire population of our purified, recombinant SMB has a solvent exposure, both as a free domain and in complex with PAI-1, which is indistinguishable from that of plasma-derived SMB as assessed by amide hydrogen ((1)H/(2)H) exchange. This solvent exposure was only reproduced by one of three synthetic SMB products with predefined disulfide connectivities corresponding to those published previously. Furthermore, this connectivity was also the only one to yield a folded and functional domain. The NMR structure was determined for free SMB produced by Pichia and is largely consistent with that solved by X-ray crystallography for SMB in complex with PAI-1.


Anatomic Double-bundle ACL Reconstruction.
Rupture of the anterior cruciate ligament (ACL) is one of the most frequent forms of knee trauma. The traditional surgical treatment for ACL rupture is single-bundle reconstruction. However, during the past few years there has been a shift in interest toward double-bundle reconstruction to closely restore the native ACL anatomy. This paper evaluates the basis for double-bundle ACL reconstruction including anatomy, biomechanics and kinematics, describes our surgical technique, and discusses why we prefer anatomic double-bundle ACL reconstruction, as well as its outcome, the choices, and the controversies of double-bundle ACL reconstruction. Pitfalls of traditional ACL surgery are also discussed, the recognition of which is the key to performing anatomic ACL reconstruction.


Flow cytometric analysis of human bone marrow: I. Normal erythroid development.
Flow cytometry was used to identify maturational differences of erythroid lineage cells in normal human bone marrow by combining physical characteristics, the expression of multiple cell surface antigens, and nucleic acid content. Normal low-density bone marrow cells could be divided into four populations, based on forward and right-angle light scattering. Erythroid cells, at different maturational stages, were found in three of these four marrow subpopulations. The sequentially correlated expression of three cell surface markers--HLe-1, transferrin receptor, and glycophorin--allowed us to study erythroid maturation from the colony forming cell to the mature erythrocyte. HLe-1 was expressed on the earliest identifiable erythroid cells and was progressively lost as the cells matured. Transferrin receptor began to be expressed at the BFU-E stage and disappeared at the late reticulocyte stage. Transferrin receptor expression preceded glycophorin expression, the latter beginning on morphologically recognizable erythroid precursors just after the CFU-E stage. In contrast to both HLe-1 and transferrin receptor, which were progressively lost during the maturational process, once glycophorin had been maximally expressed on the cell surface, it remained at constant quantities to the mature erythrocyte stage. Although developing nucleated erythroid cells at approximately the normoblast stage had light-scattering properties similar to those of lymphoid cells, these two cell types could be resolved by cell surface antigen expression. Normoblasts were glycophorin positive and HLe negative, whereas lymphoid cells expressed HLe and either Leu 4, Leu 11, or Leu 12. Decreases in cellular nucleic acid content, corresponding first to the extrusion of the nucleus and second to the loss of reticulum, characterized the later stages of erythroid development. These characteristics and instrumentation can be used to purify erythroid cells at various developmental stages.


Changes in resistance among coliform bacteraemia associated with a primary care antimicrobial stewardship intervention: A population-based interrupted time series study.
Primary care antimicrobial stewardship interventions can improve antimicrobial prescribing, but there is less evidence that they reduce rates of resistant infection. This study examined changes in broad-spectrum antimicrobial prescribing in the community and resistance in people admitted to hospital with community-associated coliform bacteraemia associated with a primary care stewardship intervention. Segmented regression analysis of data on all patients registered with a general practitioner in the National Health Service (NHS) Tayside region in the east of Scotland, UK, from 1 January 2005 to 31 December 2015 was performed, examining associations between a primary care antimicrobial stewardship intervention in 2009 and primary care prescribing of fluoroquinolones, cephalosporins, and co-amoxiclav and resistance to the same three antimicrobials/classes among community-associated coliform bacteraemia. Prescribing outcomes were the rate per 1,000 population prescribed each antimicrobial/class per quarter. Resistance outcomes were proportion of community-associated (first 2 days of hospital admission) coliform (Escherichia coli, Proteus spp., or Klebsiella spp.) bacteraemia among adult (18+ years) patients resistant to each antimicrobial/class. 11.4% of 3,442,205 oral antimicrobial prescriptions dispensed in primary care over the study period were for targeted antimicrobials. There were large, statistically significant reductions in prescribing at 1 year postintervention that were larger by 3 years postintervention when the relative reduction was -68.8% (95% CI -76.3 to -62.1) and the absolute reduction -6.3 (-7.6 to -5.2) people exposed per 1,000 population per quarter for fluoroquinolones; relative -74.0% (-80.3 to -67.9) and absolute reduction -6.1 (-7.2 to -5.2) for cephalosporins; and relative -62.3% (-66.9 to -58.1) and absolute reduction -6.8 (-7.7 to -6.0) for co-amoxiclav, all compared to their prior trends. There were 2,143 eligible bacteraemia episodes involving 2,004 patients over the study period (mean age 73.7 [SD 14.8] years; 51.4% women). There was no increase in community-associated coliform bacteraemia admissions associated with reduced community broad-spectrum antimicrobial use. Resistance to targeted antimicrobials reduced by 3.5 years postintervention compared to prior trends, but this was not statistically significant for co-amoxiclav. Relative and absolute changes were -34.7% (95% CI -52.3 to -10.6) and -63.5 (-131.8 to -12.8) resistant bacteraemia per 1,000 bacteraemia per quarter for fluoroquinolones; -48.3% (-62.7 to -32.3) and -153.1 (-255.7 to -77.0) for cephalosporins; and -17.8% (-47.1 to 20.8) and -63.6 (-206.4 to 42.4) for co-amoxiclav, respectively. Overall, there was reversal of a previously rising rate of fluoroquinolone resistance and flattening of previously rising rates of cephalosporin and co-amoxiclav resistance. The limitations of this study include that associations are not definitive evidence of causation and that potential effects of underlying secular trends in the postintervention period and/or of other interventions occurring simultaneously cannot be definitively excluded. In this population-based study in Scotland, compared to prior trends, there were very large reductions in community broad-spectrum antimicrobial use associated with the stewardship intervention. In contrast, changes in resistance among coliform bacteraemia were more modest. Prevention of resistance through judicious use of new antimicrobials may be more effective than trying to reverse resistance that has become established.


Left ventricular noncompaction cardiomyopathy: a case report and literature review.
Left ventricular noncompaction cardiomyopathy (LVNC) is a relatively rare congenital disorder prominently characterized by prominent trabeculations and intertrabecular recesses that communicate with the ventricular cavity rather than the coronary circulation. LVNC can occur in isolation or coexist with other cardiac and/or systemic anomalies, in especial neuromuscular disorders. The clinical presentation varies ranging from asymptomatic patients to patients who develop ventricular arrhythmias, thromboembolism, heart failure and sudden cardiac death. Although LVNC is commonly diagnosed by echocardiography, there are also other useful diagnostic techniques, including contrast ventriculography, CT and MRI. Now, it is being diagnosed more frequently in patients of all ages because of increased awareness and improvements in imaging methods. We described the case of a woman who presented with heart failure for the first time at 62 years of age. The diagnosis was LVNC. Transthoracic echocardiography showed a trabeculated, sponge-like appearance of the ventricular apical and inferolateral segments. After medical management, the patient was asymptomatic at the 1-month follow-up examination. Now we discussed the diagnosis of this case and reviewed the medical literature that pertained to LVNC.


Epidermal and dermal integumentary structures of ankylosaurian dinosaurs.
Ankylosaurian dinosaurs are most notable for their abundant and morphologically diverse osteoderms, which would have given them a spiky appearance in life. Isolated osteoderms are relatively common and provide important information about the structure of the ankylosaur dermis, but fossilized impressions of the soft-tissue epidermis of ankylosaurs are rare. Nevertheless, well-preserved integument exists on several ankylosaur fossils that shows osteoderms were covered by a single epidermal scale, but one or many millimeter-sized ossicles may be present under polygonal, basement epidermal scales. Evidence for the taxonomic utility of ankylosaurid epidermal scale architecture is presented for the first time. This study builds on previous osteological work that argues for a greater diversity of ankylosaurids in the Dinosaur Park Formation of Alberta than has been traditionally recognized and adds to the hypothesis that epidermal skin impressions are taxonomically relevant across diverse dinosaur clades.


Mechanisms of vascular damage by hemorrhagic snake venom metalloproteinases: tissue distribution and in situ hydrolysis.
Envenoming by viper snakes constitutes an important public health problem in Brazil and other developing countries. Local hemorrhage is an important symptom of these accidents and is correlated with the action of snake venom metalloproteinases (SVMPs). The degradation of vascular basement membrane has been proposed as a key event for the capillary vessel disruption. However, SVMPs that present similar catalytic activity towards extracellular matrix proteins differ in their hemorrhagic activity, suggesting that other mechanisms might be contributing to the accumulation of SVMPs at the snakebite area allowing capillary disruption. In this work, we compared the tissue distribution and degradation of extracellular matrix proteins induced by jararhagin (highly hemorrhagic SVMP) and BnP1 (weakly hemorrhagic SVMP) using the mouse skin as experimental model. Jararhagin induced strong hemorrhage accompanied by hydrolysis of collagen fibers in the hypodermis and a marked degradation of type IV collagen at the vascular basement membrane. In contrast, BnP1 induced only a mild hemorrhage and did not disrupt collagen fibers or type IV collagen. Injection of Alexa488-labeled jararhagin revealed fluorescent staining around capillary vessels and co-localization with basement membrane type IV collagen. The same distribution pattern was detected with jararhagin-C (disintegrin-like/cysteine-rich domains of jararhagin). In opposition, BnP1 did not accumulate in the tissues. These results show a particular tissue distribution of hemorrhagic toxins accumulating at the basement membrane. This probably occurs through binding to collagens, which are drastically hydrolyzed at the sites of hemorrhagic lesions. Toxin accumulation near blood vessels explains enhanced catalysis of basement membrane components, resulting in the strong hemorrhagic activity of SVMPs. This is a novel mechanism that underlies the difference between hemorrhagic and non-hemorrhagic SVMPs, improving the understanding of snakebite pathology.


[Dynamics of pulmonary hypertension after mitral valve prosthesis (late results)].
The presented analysis includes the haemodynamic parameters obtained during right and left heart catheterization in 51 patients following mitral valve prosthetic replacement for rheumatic mitral disease within 1 month to 8 years. In 30 patients the examination was conducted pre- and postoperatively, and in 21--postoperatively only. A comparative analysis of the pre- and postoperative haemodynamic indices has shown that when the effect of surgery is positive, the pressure in the pulmonary artery, the wedge pressure, the right atrial, and the end-diastolic pressures in the right and left ventricles decrease. A reduction is also noted in the pulmonary-arteriolar resistance, in the arterio-venous oxygen content gradient, and in the right ventricle work, and an increase in the cardiac output, cardiac index and effective work of the left ventricle. The dynamics of these parameters is especially dramatic in patients with severe pulmonary hypertension (the pulmonary artery pressure falls by 60%, and the pulmonary-arteriolar resistance--by 83%). When the operation fails to produce any haemodynamic effect, the intracardiac haemodynamics disorders persist along with those in the pulmonary circulation, which permits us to find the reason of the unsatisfactory result of surgery.


[Chemotaxonomic characteristics of bacteria belonging to the genus Nocardia isolated from Ukrainian SSR soils].
From soils of the Ukrainian SSR bearing oil and without it, 241 strains of Nocardia were isolated on mineral media containing n-alkanes C12-C22. The strains were divided into two groups: actinoid (N. asteroides, N. flavescens) and bacterium-like (N. rubropertincta, N. flava, N. ucrainica, N. salmonicolor, N. rubra, N. erythropolis, Nocardia ssp). Chemotaxonomic characteristics (the presence in the cells of galactose, arabinose, mesodiaminopimelic acid, lipid LCN-A) were used to differentiate Nocardia from other genera of actinomycetes and coryneforms and to confirm the taxonomy of museum strains. The majority of the Nocardia cultures belongs to the groups "asteroides" and "rhodochrous" according to location of spots of lipid LCN-A on chromatograms. The strains of N. asteroides and complex "M." rhodochrous may belong to both groups. A small number of strains belongs to the group which is represented by N. calcarea 41 (NCIB 8863).


Enhancement of ectopic bone formation by bone morphogenetic protein-2 delivery using heparin-conjugated PLGA nanoparticles with transplantation of bone marrow-derived mesenchymal stem cells.
This study was performed to determine if a combination of previously undifferentiated bone marrow-derived mesenchymal stem cells (BMMSCs) and exogenous bone morphogenetic protein-2 (BMP-2) delivered via heparin-conjugated PLGA nanoparticles (HCPNs) would extensively regenerate bone in vivo. In vitro testing found that the HCPNs were able to release BMP-2 over a 2-week period. Human BMMSCs cultured in medium containing BMP-2-loaded HCPNs for 2 weeks differentiated toward osteogenic cells expressing alkaline phosphatase (ALP), osteopontin (OPN) and osteocalcin (OCN) mRNA, while cells without BMP-2 expressed only ALP. In vivo testing found that undifferentiated BMMSCs with BMP-2-loaded HCPNs induce far more extensive bone formation than either implantation of BMP-2-loaded HCPNs or osteogenically differentiated BMMSCs. This study demonstrates the feasibility of extensive in vivo bone regeneration by transplantation of undifferentiated BMMSCs and BMP-2 delivery via HCPNs.


Cross-section Trichometry: A Clinical Tool for Assessing the Progression and Treatment Response of Alopecia.
To properly assess the progression and treatment response of alopecia, one must measure the changes in hair mass, which is influenced by both the density and diameter of hair. Unfortunately, a convenient device for hair mass evaluation had not been available to dermatologists until the recent introduction of the cross-section trichometer, which directly measures the cross-sectional area of an isolated bundle of hair. We sought to evaluate the accuracy and sensitivity of the HairCheck(®) device, a commercial product derived from the original cross-section trichometer. Bundles of surgical silk and human hair were used to evaluate the ability of the HairCheck(®) device to detect and measure small changes in the number and diameter of strands, and bundle weight. Strong correlations were observed between the bundle's cross-sectional area, displayed as the numeric Hair Mass Index (HMI), the number of strands, the silk/hair diameter, and the bundle dry weight. HMI strongly correlated with the number and diameter of silk/hair, and the weight of the bundle, suggesting that it can serve as a valid indicator of hair mass. We have given the name cross-section trichometry (CST) to the methodology of obtaining the HMI using the HairCheck(®) system. CST is a simple modality for the quantification of hair mass, and may be used as a convenient and useful tool to clinically assess changes in hair mass caused by thinning, shedding, breakage, or growth in males and females with progressive alopecia or those receiving alopecia treatment.


The Hidden Burden of Dengue and Chikungunya in Chennai, India.
Dengue and chikungunya are rapidly expanding viruses transmitted by mosquitoes of the genus Aedes. Few epidemiological studies have examined the extent of transmission of these infections in South India despite an increase in the number of reported cases, and a high suitability for transmission. We conducted a household-based seroprevalence survey among 1010 individuals aged 5-40 years living in fifty randomly selected spatial locations in Chennai, Tamil Nadu. Participants were asked to provide a venous blood sample and to complete a brief questionnaire with basic demographic and daily activity information. Previous exposure to dengue and chikungunya was determined using IgG indirect ELISA (Panbio) and IgG ELISA (Novatec), respectively. We used this data to estimate key transmission parameters (force of infection and basic reproductive number) and to explore factors associated with seropositivity. While only 1% of participants reported history of dengue and 20% of chikungunya, we found that 93% (95%CI 89-95%) of participants were seropositive to dengue virus, and 44% (95%CI 37-50%) to chikungunya. Age-specific seroprevalence was consistent with long-tem, endemic circulation of dengue and suggestive of epidemic chikungunya transmission. Seropositivity to dengue and chikungunya were significantly correlated, even after adjusting for individual and household factors. We estimate that 23% of the susceptible population gets infected by dengue each year, corresponding to approximately 228,000 infections. This transmission intensity is significantly higher than that estimated in known hyperendemic settings in Southeast Asia and the Americas. These results provide unprecedented insight into the very high transmission potential of dengue and chikungunya in Chennai and underscore the need for enhanced surveillance and control methods.


[The status of and problems in the development of Russian medical instrument making].
Recently, there have been great changes in the nomenclature of manufactured medical equipment (ME) items and hence there is a considerable renewal of products due to the fact that a lot of conversed enterprises of the military-industrial complex (MIC) have settled down in producing ME. By and large, the output of new ME items is 37% of their total medical instrument making nomenclature. Thus, in 1988 the proportion of the items was 35% of the total output of medical devices and apparatuses while in 1995 it is 65%. The output and technical level of some extremely complex ME items, Russian devices for hemodialysis and extracorporeal circulation have been increased. The Russian enterprises have designed and are manufacturing diagnostic nuclear magnetic systems, pulse oximeters, Billitest bilirubin meters, complex laboratory sets for radioimmunochemical assay, a G202-5 lit-par-lit scanning complex, Mammodiagnost apparatus, new Roentgen-48 and Roentgen-60 X-ray complexes, etc. The paper gives specific examples how a number of medical devices and apparatuses are designed and launched into production at the electronic and radio industrial and MIC enterprises.


Attention Modification to Attenuate Facial Emotion Recognition Deficits in Children with Autism: A Pilot Study.
Diminished attending to faces may contribute to the impairments in emotion recognition and expression in autism spectrum disorder (ASD). The current study evaluated the acceptability, feasibility, and preliminary efficacy of an attention modification intervention designed to attenuate deficits in facial emotion recognition (FER). During the 10-session experimental treatment, children (n = 8) with ASD watched dynamic videos of people expressing different emotions with the facial features highlighted to guide children's attention. Children and their parents generally rated the treatment as acceptable and helpful. Although FER improvement was not apparent on task-based measures, parents reported slight improvements and decreased socioemotional problems following treatment. Results suggest that further research on visual attention retraining for ASD, within an experimental therapeutic program, may be promising.


Modification of the beta- and alpha2-adrenergic sensitivity of rat submandibular glands by environmental stimuli and stress.
In man, the rate of resting salivary secretion can be influenced by environmental stimuli related to light dark cycles or by noxious stimuli (stressors) of psychological origin. The sympathetic branch of the autonomic nervous system and the adrenal medulla play an important part in homeostatic responses. Previous observations have shown that chronic exposure of rats to constant light promotes degranulation of parotid acini and desensitization of submandibular beta-adrenergic receptors. Now the submandibular secretory response elicited by beta- and alpha2-adrenergic agonists was studied in rats chronically exposed to environmental conditions that modified the activities of sympathetic efferents to the pineal, salivary and adrenal glands. Adult male rats were exposed to constant light (LL) or constant darkness (DD) for 20 days, or to stress (2 h daily immobilization) for 14 days. Control animals were kept under the usual lighting conditions and without immobilization. Dose response curves to isoproterenol (i.v), before and after administration (i.v.) of a dose (20 microg/kg) of clonidine were obtained. Beta-adrenergic desensitization was observed in all the experimental groups, while alpha2-adrenergic desensitization was only observed in the stress and LL groups. The results suggest that circulating catecholamines could mediate light and stress effects on submandibular beta-adrenergic secretory responses. Extrasynaptic alpha2-adrenoceptors might modulate the submandibular secretory response when predictable environmental stimuli (daily light phase) or unpredictable stressors raise the concentrations of circulating catecholamines.


Young adult oncology: the patients and their survival challenges.
One in every 168 Americans develops invasive cancer between age 15 to 30 years. During this age interval, cancer is unique in the distribution of types that occur and rarely related to either environmental carcinogens, a recognizable inherited predisposition, or a family cancer syndrome. Patients in this age group have the lowest rate of health insurance coverage, frequent delays in diagnosis, and the lowest accrual to clinical trials. Their psychosocial needs are unique and generally less well attended to than in any other age group. Despite an intrinsically equal ability to tolerate chemotherapy, older adolescents and young adults frequently receive lower dose intensities than do younger patients, and at times less than in older patients. Whereas the 15- to 29-year age group once had a better overall survival rate than either younger or older patients, a relative lack of progress has resulted in the majority of cancers in the age group having a worse overall survival rate than in younger patients, and several of these having a worse prognosis than in older patients. Against this background, young adults with cancer have unique survival challenges--medically, psychosocially, and economically--that are now beginning to be appreciated and addressed with a national initiative.


Prognostic value of dynamic electrocardiographic T wave changes in non-ST elevation acute coronary syndrome.
To assess the relationship between the evolution of T wave inversion (TWI) on the 24-48 h postadmission ECG and the patient characteristics, management and clinical outcomes among those with non-ST elevation acute coronary syndrome (NSTE-ACS). We evaluated admission and 24-48 h follow-up ECGs of 7201 patients with NSTE-ACS from the prospective, multicentre Global Registry of Acute Coronary Events (GRACE) and Canadian ACS Registry I. We performed multivariable analyses to determine the association between new TWI (on follow-up ECG only), resolved TWI (on admission ECG only) and persistent TWI (on both admission and follow-up ECG) and inhospital and cumulative 6-month all-cause mortality. Patients with TWI were older, more likely to have cardiovascular risk factors, higher Killip class and GRACE risk scores. After adjustment for known prognostic factors, compared with patients presenting without TWI, new TWI was associated with significantly lower inhospital mortality (OR=0.60, 95% CI 0.38 to 0.95, p=0.029), whereas resolved (OR=1.06, 95% CI 0.65 to 1.75, p=0.81) and persistent (OR=0.73, 95% CI 0.48 to 1.11, p=0.14) TWI did not predict inhospital mortality. No TWI pattern independently predicted inhospital adverse cardiovascular events or cumulative 6-month mortality. In contrast, ST depression on the admission and follow-up ECG were independent predictors of inhospital and 6-month mortality. Across the spectrum of NSTE-ACS, TWI within 48 h of presentation was associated with high-risk clinical features, but its presence or dynamic change did not provide additional prognostic value beyond other established clinical predictors.


Photodegradation of melanin by an interferometric technique.
Photodegradation of melanin thin films is investigated for a UVA wavelength of 355 nm and a UVC wavelength of 244 nm. The technique involves interferometric exposure of melanin with two coherent beams from a low-power UV laser. The periodic photodegradation-grating pattern is monitored by diffraction of a second low-power He-Ne laser. Dependence of the photodegradation rate on UV intensity as well as the effect of ambient humidity is investigated and explained with a simple model. The technique has promise for investigating photo-induced effects in other biomolecular substrates as well.


Balloon pulmonary angioplasty for chronic thromboembolic pulmonary hypertension due to septic emboli.
We present the case of a 28-year-old woman with a history of tricuspid valve endocarditis leading to chronic thromboembolic pulmonary hypertension (CTEPH) with multiple pulmonary artery chronic total occlusions (CTOs) due to septic emboli. Following a multidisciplinary care discussion, the patient was brought forward for balloon pulmonary angioplasty (BPA) with successful revascularization of all chronically occluded territories. This case highlights advances in pulmonary artery CTO interventions and demonstrates the feasibility of BPA for CTEPH patients with a history of septic emboli.


Lipoprotein(a) is robustly associated with aortic valve calcium.
To investigate the prevalence and quantity of aortic valve calcium (AVC) in two large cohorts, stratified according to age and lipoprotein(a) (Lp(a)), and to assess the association between Lp(a) and AVC. We included 2412 participants from the population-based Rotterdam Study (52% women, mean age=69.6±6.3 years) and 859 apparently healthy individuals from the Amsterdam University Medical Centers (UMC) outpatient clinic (57% women, mean age=45.9±11.6 years). All individuals underwent blood sampling to determine Lp(a) concentration and non-enhanced cardiac CT to assess AVC. Logistic and linear regression analyses were performed to investigate the associations of Lp(a) with the presence and amount of AVC. The prevalence of AVC was 33.1% in the Rotterdam Study and 5.4% in the Amsterdam UMC cohort. Higher Lp(a) concentrations were independently associated with presence of AVC in both cohorts (OR per 50 mg/dL increase in Lp(a): 1.54 (95% CI 1.36 to 1.75) in the Rotterdam Study cohort and 2.02 (95% CI 1.19 to 3.44) in the Amsterdam UMC cohort). In the Rotterdam Study cohort, higher Lp(a) concentrations were also associated with increase in aortic valve Agatston score (β 0.19, 95% CI 0.06 to 0.32 per 50 mg/dL increase). Lp(a) is robustly associated with presence of AVC in a wide age range of individuals. These results provide further rationale to assess the effect of Lp(a) lowering interventions in individuals with early AVC to prevent end-stage aortic valve stenosis.


Assessing outcomes in liver disease patients: reliability and validity of the Spanish version of the Liver Disease Quality of Life Questionnaire (LDQOL 1.0).
To assess the reliability and validity of a Spanish version of the LDQOL 1.0 (Liver Disease Quality of Life questionnaire). Observational, cross-sectional study in Spanish patients awaiting liver transplantation (LT). Feasibility was assessed by analyzing administration times and missing responses. Ceiling and floor effects were calculated and reliability was tested by examining internal consistency (Cronbach's alpha). Convergent validity was tested by examining correlations between LDQOL disease-specific and Short Form health survey with 36 questions (SF-36) dimensions. Known groups' validity was tested by examining the LDQOL's capacity to discriminate between groups defined by etiology and Child-Turcotte-Pugh (CTP) scores. A total of 200 patients were included for analysis. Mean age (SD) was 52.6 (9.8) years and 73% of the sample were male. The most common indication for LT was liver cancer (34%). Mean (SD) time to complete the questionnaire was 35.8 minutes (21.2 minutes). Missing responses were highest on the dimensions of sexual functioning and symptoms of liver disease. Ceiling effects were over 20% on 7 of the LDQOL's 12 disease-specific scales. Cronbach's alpha coefficients were over 0.70 on all but 2 dimensions. Correlations between SF-36 and LDQOL disease-specific dimensions generally fulfilled the hypotheses, with 35 of the 40 highest and lowest correlations (87.5%) being in the expected direction. The LDQOL discriminated well between patients in CTP class A and C, and as hypothesized, hepatocarcinoma and alcoholic cirrhosis patients scored better on most dimensions than patients with hepatitis C virus or other etiologies. The Spanish version of the LDQOL 1.0 has shown satisfactory reliability and validity.


Quantitative electron microscopy and fluorescence spectroscopy of the membrane distribution of influenza hemagglutinin.
Although lipid-dependent protein clustering in biomembranes mediates numerous functions, there is little consensus among membrane models on cluster organization or size. Here, we use influenza viral envelope protein hemagglutinin (HA(0)) to test the hypothesis that clustering results from proteins partitioning into preexisting, fluid-ordered "raft" domains, wherein they have a random distribution. Japan HA(0) expressed in fibroblasts was visualized by electron microscopy using immunogold labeling and probed by fluorescence resonance energy transfer (FRET). Labeled HA coincided with electron-dense, often noncircular membrane patches. Poisson and K-test (Ripley, B.D. 1977. J. R. Stat. Soc. Ser. B. 39:172-212) analyses reveal clustering on accessible length scales (20-900 nm). Membrane treatments with methyl-beta-cyclodextrin and glycosphingolipid synthesis inhibitors did not abolish clusters but did alter their pattern, especially at the shortest lengths, as was corroborated by changes in FRET efficiency. The magnitude and density dependence of the measured FRET efficiency also indicated a nonrandom distribution on molecular length scales (approximately 6-7 nm). This work rules out the tested hypothesis for HA over the accessible length scales, yet shows clearly how the spatial distribution of HA depends on lipid composition.


Correlation between Fc γ R III a and aortic atherosclerotic plaque destabilization in ApoE knockout mice and intervention effects of effective components of chuanxiong rhizome and red peony root.
To explore the correlation between Fc γ RIII A (CD16A) and aortic atherosclerotic plaque destabilization in apoE knockout (apoE KO) mice and the intervention effects of effective components of chuanxiong rhizome and red peony root. Eight 8-week-old male C57BL/6J mice were selected as the control group. Forty 8-week-old male apoE KO mice were randomly divided into the model group, apoE KO + intraperitoneal injection immunoglobulin group (IVIG), apoE KO + simvastatin group (Sm), apoE KO + high dosage of xiongshao capsule (XSC) group (XSCH), and apoE KO + low dosage of XSC group (XSCL), 8 mice in each group. Mice in the control group were put on a normal diet, and others were fed with a high-fat diet. After 10-week different interventions, monocyte CD16 expression was detected by flow cytometry, aortic matrix metalloproteinase-9 (MMP-9) mRNA expression was detected using reverse transcription polymerase chain reaction, and serum tumor necrosis factor (TNF)-α level was detected using enzyme-linked immunosorbent assay. Compared with the control group, monocyte CD16 expression, aortic MMP-9 mRNA expression, and serum TNF-α level in the model group increased obviously (P<0.01). Injections of apoE KO mice with intraperitoneal immunoglobulin during a 5-day period significantly reduced the monocyte CD16 expression, aortic MMP-9 mRNA expression, and serum TNF-α level (P<0.01 or 0.05) over a 10-week period of high-fat diet. Indices above in the Sm group, XSCH group, and XSCL group decreased in a different degree. Of them, the aortic MMP-9 mRNA expression in XSCH group was lower than that in Sm group (P<0.05) and the monocyte CD16 expression and serum TNF-α level showed no significant difference between XSCH group and Sm group (P>0.05). Correlation analyses suggested positive correlation between monocyte CD16 expression and aortic MMP-9 mRNA expression or serum TNF-α level in IVIG group, XSCH group, and XSCL group. FcγR III A mediates systemic inflammation in the progression of coronary heart disease with blood stasis syndrome. XSC could stabilize atherosclerotic plaque by suppressing inflammation and its target was relative with FcγRIII A.


Radiation protection practices of staff during extra-corporeal shock wave lithotripsy at Okada, Nigeria.
The study was designed to find out the radiation protection practices of radiologists and other staff involved in the first extra-corporeal shock wave lithotripsy in Nigeria, performed at Igbinedion Hospital and Medical Research Centre, Okada. Some members of staff who were present when the extra-corporeal shock wave lithotripsy (ESWL) was used in the hospital at Okada were interviewed between November 2002 and August 2003. Radiology records of the hospital were studied. Literature search involved available publication on the procedure in local and international journals with interest in precautions to reducing radiation exposure. Only lead apron and lead gloves were used by the radiologists for radiation protection and shielding during fluoroscopy procedures. The fluoroscopy was the screen type with TV monitor. Multiple sessions were used in several patients with multiple pre- and post- treatment radiographic studies including contrast studies with average of two sessions per patient. All the patients were adults aged between 26 and 65 years with mean age of 42.5 years. 627-6000 shock waves were delivered over 45-135 minutes at intensity of 143-19KV depending on patients build and the size of the stones. The sizes of the patients varied from very obese with large bulk to slim built. Radiation monitoring of the staff and patients was not done. Staff believed that radiation effect from the lithotripsy procedure was low therefore adequate radiation monitoring and radiation reducing alteration in the procedure was rarely adopted. Extended fluoroscopy time, multiple fluoroscopy examinations, multiple treatment sessions and multiple x-ray examinations which increased both the patients' and staff's radiation exposures were noted. Proper radiation protection and monitoring of patients and staff are necessary to avoid the risks from low-level exposure to radiation such as in ESWL.


[Implementation of the academic day for undergraduate interns].
Medical internship is an important period in the formation of a general (or medical) practitioner, in which the student becomes a member of the "health community" and also applies all the knowledge he or she has received during the university lectures. The present study aims to evaluate the impact of the implementation of the academic day programme designed for undergraduate medical interns within the Mexican second level healthcare Hospital General Regional No. 6 "Dr. Ignacio García Téllez". A comparative and also a prospective study were applied to a sample of 56 medical interns who participated in the academic day implementation. The impact of the scheme was assessed using the notes of the different exams at the beginning and end of the medical internship. The non-parametrical Wilcoxon test was used for the statistical analysis for comparison between the initial and final notes. A positive difference was observed in the notes of the modules of internal medicine, pediatrics, gynecology, emergency, general surgery, and family medicine applied at the beginning and end of the implementation of the academic day (p=0.000). Basic guidelines should be established for educational training from an active-participative perspective as the academic day scheme; these rules should significatively increase the medical intern learning process and, as a logical consequence would have a positive influence on healthcare quality.


3-T imaging of the cochlear nerve and labyrinth in cochlear-implant candidates: 3D fast recovery fast spin-echo versus 3D constructive interference in the steady state techniques.
High-resolution imaging of the internal auditory canal and labyrinth at 1.5 T is often performed by using three-dimensional (3D) fast spin-echo or T2* techniques. We evaluated both techniques at 3 T in the preoperative assessment of patients being considered for cochlear implants. Sagittal 3D fast recovery fast spin-echo (FRFSE) and 3D constructive interference in the steady state (CISS) images were acquired in eight patients at 3.0 T by using dual surface coils. Contrast-to-noise ratios (CNRs) for the intracanalicular nerve and CSF were measured in the internal auditory canal. Two neuroradiologists reviewed the images to determine whether the techniques provided images of diagnostic quality. CNRs for 3D CISS were twice those obtained with 3D FRFSE. Both techniques provided images of diagnostic quality, though spurious signal intensity loss at the apex of the superior semicircular canals was encountered on 3D FRFSE images in four of eight patients. Both 3D FRFSE and 3D CISS provide high-resolution images of the internal auditory canal and labyrinth at 3.0 T. We predict that the superior CNRs obtained with 3D CISS will prove advantageous as we move to smaller fields of view at higher field strength.


Signals of heterogeneous selection at an MHC locus in geographically proximate ecotypes of sockeye salmon.
The genes of the major histocompatibility complex (MHC) are an important component of the vertebrate immune system and can provide insights into the role of pathogen-mediated selection in wild populations. Here, we examined variation at the MHC class II peptide-binding region in 27 populations of sockeye salmon (Oncorhynchus nerka), distributed among three distinct spawning ecotypes, from a complex of interconnected rivers and lakes in south-western Alaska. We also obtained genotypes from 90 putatively neutral single nucleotide polymorphisms for each population to compare the relative roles of demography and selection in shaping the observed MHC variation. We found that MHC divergence was generally partitioned by spawning ecotype (lake beaches, rivers and streams) and was 30 times greater than variation at neutral markers. Additionally, we observed substantial differences in modes of selection and diversity among ecotypes, with beach populations displaying higher levels of directional selection and lower MHC diversity than the other two ecotypes. Finally, the level of MHC differentiation in our study system was comparable to that observed over much larger geographic ranges, suggesting that MHC variation does not necessarily increase with increasing spatial scale and may instead be driven by fine-scale differences in pathogen communities or pathogen virulence. The low levels of neutral structure and spatial proximity of populations in our study system indicate that MHC differentiation can be maintained through strong selective pressure even when ample opportunities for gene flow exist.


The role of curvature feedback in the energetics and dynamics of lamprey swimming: A closed-loop model.
Like other animals, lampreys have a central pattern generator (CPG) circuit that activates muscles for locomotion and also adjusts the activity to respond to sensory inputs from the environment. Such a feedback system is crucial for responding appropriately to unexpected perturbations, but it is also active during normal unperturbed steady swimming and influences the baseline swimming pattern. In this study, we investigate different functional forms of body curvature-based sensory feedback and evaluate their effects on steady swimming energetics and kinematics, since little is known experimentally about the functional form of curvature feedback. The distributed CPG is modeled as chains of coupled oscillators. Pairs of phase oscillators represent the left and right sides of segments along the lamprey body. These activate muscles that flex the body and move the lamprey through a fluid environment, which is simulated using a full Navier-Stokes model. The emergent curvature of the body then serves as an input to the CPG oscillators, closing the loop. We consider two forms of feedback, each consistent with experimental results on lamprey proprioceptive sensory receptors. The first, referred to as directional feedback, excites or inhibits the oscillators on the same side, depending on the sign of a chosen gain parameter, and has the opposite effect on oscillators on the opposite side. We find that directional feedback does not affect beat frequency, but does change the duration of muscle activity. The second feedback model, referred to as magnitude feedback, provides a symmetric excitatory or inhibitory effect to oscillators on both sides. This model tends to increase beat frequency and reduces the energetic cost to the lamprey when the gain is high and positive. With both types of feedback, the body curvature has a similar magnitude. Thus, these results indicate that the same magnitude of curvature-based feedback on the CPG with different functional forms can cause distinct differences in swimming performance.


The effect of eugenol anesthesia on the electric organ discharge of the weakly electric fish Apteronotus leptorhynchus.
Eugenol, the major active ingredient of clove oil, is widely used for anesthesia in fish. Yet virtually nothing is known about its effects on CNS functions, and thus about potential interference with neurophysiological experimentation. To address this issue, we employed a neuro-behavioral assay recently developed for testing of water-soluble anesthetic agents. The unique feature of this in-vivo tool is that it utilizes a readily accessible behavior, the electric organ discharge (EOD), as a proxy of the neural activity generated by a brainstem oscillator, the pacemaker nucleus, in the weakly electric fish Apteronotus leptorhynchus. A deep state of anesthesia, as assessed by the cessation of locomotor activity, was induced within less than 3 min at concentrations of 30-60 µL/L eugenol. This change in locomotor activity was paralleled by a dose-dependent, pronounced decrease in EOD frequency. After removal of the fish from the anesthetic solution, the frequency returned to baseline levels within 30 min. Eugenol also led to a significant increase in the rate of 'chirps,' specific amplitude/frequency modulations of the EOD, during the 30 min after the fish's exposure to the anesthetic. At 60 µL/L, eugenol induced a collapse of the EOD amplitude after about 3.5 min in half of the fish tested. The results of our study indicate strong effects of eugenol on CNS functions. We hypothesize that these effects are mediated by the established pharmacological activity of eugenol to block the generation of action potentials and to reduce the excitability of neurons; as well as to potentiate GABAA-receptor responses.


Infection control guidelines for prevention of health care-associated transmission of hepatitis B and C viruses.
Viral hepatitis was first identified as an occupational hazard for health care workers more than 60 years ago. For the past few decades, hepatitis B has been one of the most significant occupational infectious risks for health care providers. With the increasing prevalence of hepatitis C infections around the world, occupational transmission of this flavivirus from infected patients to their providers has also become a significant concern. Several factors influence the risk for occupational blood-borne hepatitis infection among health care providers, among them: the prevalence of infection among the population served, the infection status of the patients to whom workers are exposed (ie, the source patient's circulating viral burden), the types and frequencies of parenteral and mucosal exposures to blood and blood-containing body fluids, and whether the patient or provider has been immunized with the hepatitis B vaccine. This article reviews patient-to-provider, patient-to-patient, and provider-to-patient transmission of hepatitis B and C in the health care setting. Current prevention strategies, precautions, and guidelines are discussed.


A Novel Splice-site Allelic Variant is Responsible for Wilson Disease in an Omani Family.
The objective of this study was to characterise Wilson's Disease (WD) [OMIM 277900] genetically and test for allelic variants in the copper transport gene (ATPase, Cu(++) transporting, beta polypeptide, ATP7B) responsible for the disease in an Omani family. Three index patients from an Omani family had been previously diagnosed with WD. All three patients suffered neurological symptoms and signs. Forty-six relatives in the family were screened for WD. Eleven more individuals were positive, but asymptomatic. Thirteen non-disease-causing allelic gene variants, described previously, were identified in the ATP7B gene from 46 family members. A putative novel disease-causing splice-site variant (c.2866-2A>G), which has not been reported previously, was detected in this family. It is located upstream of exon 13 which encodes part of transmembrane copper channel (Ch/Tm6). Reverse transcription polymerase chain reaction was used to amplify a complementary DNA (cDNA) fragment containing exons 12, 13 and 14. Exon 13 was entirely skipped from the transcript which probably would result in a defective ATP7B protein. A new ATP7B splice-site allelic variant, found among the 14 WD patients segregated with the disease in a recessive manner, suggests it is a disease-causing variant.


Use of a high-throughput screen to identify Leptospira mutants unable to colonize the carrier host or cause disease in the acute model of infection.
The molecular basis for leptospirosis infection and colonization remains poorly understood, with no efficient methods available for screening libraries of mutants for attenuation. We analysed the attenuation of leptospiral transposon mutants in vivo using a high-throughput method by infecting animals with pooled sets of transposon mutants. A total of 95 mutants was analysed by this method in the hamster model of acute infection, and one mutant was identified as attenuated (M1233, lb058 mutant). All virulence factors identified in Leptospira to date have been characterized in the acute model of infection, neglecting the carrier host. To address this, a BALB/c mouse colonization model was established. The lb058 mutant and two mutants defective in LPS synthesis were colonization deficient in the mouse model. By applying the high-throughput screening method, a further five colonization-deficient mutants were identified for the mouse model; these included two mutants in genes encoding proteins with a predicted role in iron uptake (LB191/HbpA and LB194). Two attenuated mutants had transposon insertions in either la0589 or la2786 (encoding proteins of unknown function). The final attenuated mutant had an unexpected deletion of genes la0969-la0975 at the point of transposon insertion. This is the first description of defined, colonization-deficient mutants in a carrier host for Leptospira. These mutants were either not attenuated or only weakly attenuated in the hamster model of acute leptospirosis, thus illustrating that different factors that may be required in the carrier and acute models of leptospiral infection. High-throughput screening can reduce the number of animals used in virulence studies and increase the capacity to screen mutants for attenuation, thereby enhancing the likelihood of detecting unique virulence factors. A comparison of virulence factors required in the carrier and acute models of infection will help to unravel colonization and dissemination mechanisms of leptospirosis.


Hypercoagulability in End-stage Liver Disease: Review of Epidemiology, Etiology, and Management.
In this review, we analyze the epidemiology of thromboses related to end-stage liver disease (ESLD), discuss causes of hypercoagulability, describe susceptible populations, and critically evaluate proposed prophylaxis and treatment of thromboses. Classically, ESLD has been regarded as a model for coagulopathy, and patients were deemed to be at high risk for bleeding complications. Patients with ESLD are not auto-anticoagulated, and they do not have a lower risk of portal vein thrombosis, intracardiac thrombus formation, pulmonary embolism or hepatic artery thrombosis. Though the cause of hypercoagulability is multifactorial, endothelial dysfunction likely plays a central role for all patients with ESLD. Some subpopulations, such as patients with nonalcoholic steatohepatitis and autoimmune conditions, are at increased risk of thrombotic events as are patients of Hispanic ethnicity. The science behind prophylaxis of different types of clotting and treatment of thromboses is developing rapidly. A number of medications, including low molecular weight heparin, unfractionated heparin, aspirin, vitamin K antagonists, and direct oral anticoagulants can be used, but clear guidelines are lacking. Acute intraoperative clotting can be associated with high mortality. Routine use of transesophageal echocardiography can be helpful in early recognition and treatment of intraoperative thrombosis. Heparin should be reserved for cases of intracardiac thrombus/pulmonary embolism without hemodynamic instability. In unstable patients, low dose of recombinant tissue plasminogen activator can be used. In this new era of heightened awareness of thrombotic events in ESLD patients, prospective randomized trials are urgently needed to best guide clinical practice.


Distributive justice and the harm to medical professionals fighting epidemics.
The exposure of doctors, nurses and other medical professionals to risks in the context of epidemics is significant. While traditional medical ethics offers the thought that these dangers may limit the extent to which a duty to care is applicable in such situations, it has less to say about what we might owe to medical professionals who are disadvantaged in these contexts. Luck egalitarianism, a responsibility-sensitive theory of distributive justice, appears to fare particularly badly in that regard. If we want to maintain that medical professionals are responsible for their decisions to help, cure and care for the vulnerable, luck egalitarianism seems to imply that their claim of justice to medical attention in case of infection is weak or non-existent. The article demonstrates how a recent interpretation of luck egalitarianism offers a solution to this problem. Redefining luck egalitarianism as concerned with responsibility for creating disadvantages, rather than for incurring disadvantage as such, makes it possible to maintain that medical professionals are responsible for their choices and that those infected because of their choice to help fight epidemics have a full claim of justice to medical attention.


Impact of evidence-based standardized assessment on the disability clinical interview for diagnosis of service-connected PTSD: a cluster-randomized trial.
Posttraumatic stress disorder (PTSD) is one of the fastest growing compensated medical conditions. The present study compared usual disability examiner practices for PTSD with a standardized assessment that incorporates evidence-based assessments. The design was a multicenter, cluster randomized, parallel-group study involving 33 clinical examiners and 384 veterans at 6 Veterans Affairs medical centers. The standardized group incorporated the Clinician Administered PTSD Scale and the World Health Organization Disability Assessment Schedule-II into their assessment interview. The main outcome measures were completeness and accuracy of PTSD diagnosis and completeness of functional assessment. The standardized assessments were 85% complete for diagnosis compared to 30% for nonstandardized assessments (p < .001), and, for functional impairment, 76% versus 3% (p < .001). The findings demonstrate that the quality of PTSD disability examination would be improved by using evidence-based assessment.


Habituation to thaxtomin A in hybrid poplar cell suspensions provides enhanced and durable resistance to inhibitors of cellulose synthesis.
Thaxtomin A (TA), a phytotoxin produced by the phytopathogen Streptomyces scabies, is essential for the development of potato common scab disease. TA inhibits cellulose synthesis but its actual mode of action is unknown. Addition of TA to hybrid poplar (Populus trichocarpa x Populus deltoides) cell suspensions can activate a cellular program leading to cell death. In contrast, it is possible to habituate hybrid poplar cell cultures to grow in the presence of TA levels that would normally induce cell death. The purpose of this study is to characterize TA-habituated cells and the mechanisms that may be involved in enhancing resistance to TA. Habituation to TA was performed by adding increasing levels of TA to cell cultures at the time of subculture over a period of 12 months. TA-habituated cells were then cultured in the absence of TA for more than three years. These cells displayed a reduced size and growth compared to control cells and had fragmented vacuoles filled with electron-dense material. Habituation to TA was associated with changes in the cell wall composition, with a reduction in cellulose and an increase in pectin levels. Remarkably, high level of resistance to TA was maintained in TA-habituated cells even after being cultured in the absence of TA. Moreover, these cells exhibited enhanced resistance to two other inhibitors of cellulose biosynthesis, dichlobenil and isoxaben. Analysis of gene expression in TA-habituated cells using an Affymetrix GeneChip Poplar Genome Array revealed that durable resistance to TA is associated with a major and complex reprogramming of gene expression implicating processes such as cell wall synthesis and modification, lignin and flavonoid synthesis, as well as DNA and chromatin modifications. We have shown that habituation to TA induced durable resistance to the bacterial toxin in poplar cells. TA-habituation also enhanced resistance to two other structurally different inhibitors of cellulose synthesis that were found to target different proteins. Enhanced resistance was associated with major changes in the expression of numerous genes, including some genes that are involved in DNA and chromatin modifications, suggesting that epigenetic changes might be involved in this process.


Prevalence and associated factors of poly-victimization in Chinese adolescents.
Multiple types of victimization or poly-victimization (PV) can occur simultaneously during childhood, resulting in outcomes that are detrimental and difficult to reverse. Very few studies have focused on PV in Chinese adolescents. The present study is based on information collected from a stratified cluster sample of 1,561 females and 1,594 males aged 12-18 years living in Shandong Province, China. A self-administered questionnaire was used to assess background information, the Juvenile Victimization Questionnaire was used to measure the extent of victimization and PV, the Screen for Child Anxiety Related Emotional Disorders was used to measure anxiety levels, and a Depression Self-rating Scale for Children was used to ascertain depression. A majority of the study sample (66.2%) had experienced at least one form of victimization over the past year. Prevalence of PV was 16.9%, and was significantly higher among males (21.1%) than females (12.5%). Younger ages, one-child families, and lower socioeconomic status were positively associated with PV. Logistic regression analysis indicated that smoking, alcohol consumption, exposure to pornography, and anxiety- and depression-like symptoms were risk factors of PV. Results of this study highlight the need for further exploration of factors related to the PV of Chinese adolescents.


One out of four recruits drops out from elite military training due to musculoskeletal injuries in the Netherlands Armed Forces.
Musculoskeletal injuries (MSIs) are among the main causes of dropout from military training. The main purpose of this study was to provide an overview of dropout rates and MSI incidence rates during elite military training. Second, this study aimed to explore restricted training days due to MSIs and to describe MSI-care by military physicians. In a retrospective observational study, we collected dropout rates and injury surveillance data from the electronic patient records of two elite units of the Netherlands Armed Forces (NAF): the Royal Netherlands Marine Corps (RNLMC) and the Airmobile Brigade (AMB), from 1 January 2015 until 31 December 2017. In the RNLMC, total dropout rate was 53.9% and dropout due to MSIs was 23%. The most frequently affected locations were foot, knee and leg. In the AMB total dropout rate was 52.6% and dropout due to MSIs was 25%. In the AMB, the most frequently affected locations were back, knee and leg. Average restricted training days due to MSIs ranged between 8.3 and 20.8 days/injury. MSI-care by military physicians consisted mostly of the provision of injury-specific information and (self-)management options, imposing a specific activity restriction and referral to physiotherapy. Our study findings showed that one out of four recruits who dropout from elite military training in the NAF, do so due to MSIs. Redesigning training programmes with the objective to reduce MSIs should be given high priority, as this may reduce dropout substantially.


Lymphocyte recognition of high endothelium: antibodies to distinct epitopes of an 85-95-kD glycoprotein antigen differentially inhibit lymphocyte binding to lymph node, mucosal, or synovial endothelial cells.
The tissue-specific homing of lymphocytes is directed by specialized high endothelial venules (HEV). At least three functionally independent lymphocyte/HEV recognition systems exist, controlling the extravasation of circulating lymphocytes into peripheral lymph nodes, mucosal lymphoid tissues (Peyer's patches or appendix), and the synovium of inflamed joints. We report here that antibodies capable of inhibiting human lymphocyte binding to one or more HEV types recognize a common 85-95-kD lymphocyte surface glycoprotein antigen, defined by the non-blocking monoclonal antibody, Hermes-1. We demonstrate that MEL-14, a monoclonal antibody against putative lymph node "homing receptors" in the mouse, functionally inhibits human lymphocyte binding to lymph node HEV but not to mucosal or synovial HEV, and cross-reacts with the 85-95-kD Hermes-1 antigen. Furthermore, we show that Hermes-3, a novel antibody produced by immunization with Hermes-1 antigen isolated from a mucosal HEV-specific cell line, selectively blocks lymphocyte binding to mucosal HEV. Such tissue specificity of inhibition suggests that MEL-14 and Hermes-3 block the function of specific lymphocyte recognition elements for lymph node and mucosal HEV, respectively. Recognition of synovial HEV also involves the 85-95-kD Hermes-1 antigen, in that a polyclonal antiserum produced against the isolated antigen blocks all three classes of lymphocyte-HEV interaction. From these studies, it is likely that the Hermes-1-defined 85-95-kD glycoprotein class either comprises a family of related but functionally independent receptors for HEV, or associates both physically and functionally with such receptors. The findings imply that related molecular mechanisms are involved in several functionally independent cell-cell recognition events that direct lymphocyte traffic.


The hepatokine Tsukushi is released in response to NAFLD and impacts cholesterol homeostasis.
Nonalcoholic fatty liver disease (NAFLD) prevails in obesity and is linked to several health complications including dyslipidemia and atherosclerosis. How exactly NAFLD induces atherogenic dyslipidemia to promote cardiovascular diseases is still elusive. Here, we identify Tsukushi (TSK) as a hepatokine induced in response to NAFLD. We show that both endoplasmic reticulum stress and inflammation promote the expression and release of TSK in mice. In humans, hepatic TSK expression is also associated with steatosis, and its circulating levels are markedly increased in patients suffering from acetaminophen-induced acute liver failure (ALF), a condition linked to severe hepatic inflammation. In these patients, elevated blood TSK levels were associated with decreased transplant-free survival at hospital discharge, suggesting that TSK could have a prognostic significance. Gain- and loss-of-function studies in mice revealed that TSK impacts systemic cholesterol homeostasis. TSK reduces circulating HDL cholesterol, lowers cholesterol efflux capacity, and decreases cholesterol-to-bile acid conversion in the liver. Our data identify the hepatokine TSK as a blood biomarker of liver stress that could link NAFLD to the development of atherogenic dyslipidemia and atherosclerosis.


Quantitative genetic analysis of salicylic acid perception in Arabidopsis.
Salicylic acid (SA) is a phytohormone required for a full resistance against some pathogens in Arabidopsis, and NPR1 (Non-Expressor of Pathogenesis Related Genes 1) is the only gene with a strong effect on resistance induced by SA which has been described. There can be additional components of SA perception that escape the traditional approach of mutagenesis. An alternative to that approach is searching in the natural variation of Arabidopsis. Different methods of analyzing the variation between ecotypes have been tried and it has been found that measuring the growth of a virulent isolate of Pseudomonas syringae after the exogenous application of SA is the most effective one. Two ecotypes, Edi-0 and Stw-0, have been crossed, and their F2 has been studied. There are two significant quantitative trait loci (QTLs) in this population, and there is one QTL in each one of the existing mapping populations Col-4 × Laer-0 and Laer-0 × No-0. They have different characteristics: while one QTL is only detectable at low concentrations of SA, the other acts after the point of crosstalk with methyl jasmonate signalling. Three of the QTLs have candidates described in SA perception as NPR1, its interactors, and a calmodulin binding protein.


Improving outpatient warfarin use for hospitalized patients with atrial fibrillation.
Atrial fibrillation affects an estimated 5 million Americans and accounts for approximately 15% of all strokes. Few studies have successfully addressed patient screening, assessment, and introduction of appropriate antithrombotic therapy in patients with atrial fibrillation. To assess whether an intervention improved planned antithrombotic prescribing at the time of discharge in hospitalized patients. The study was a prospectively designed, retrospectively evaluated, non-blinded, historical control study of a pharmacist-initiated intervention. The intervention, consisting of pharmacist review and assessment of antithrombotic prescribing in patients with non-valvular atrial fibrillation, was conducted in an urban teaching hospital. Although antithrombotic prescribing was not significantly higher at discharge in the 252 enrolled subjects (control 67.3% vs. intervention 70.8%; p = 0.58), a significantly greater number of patients had a written discharge plan for antithrombotic therapy (control 73.5% vs. intervention 88.3%; p < 0.01). The adjusted odds ratio that the study group was associated with an improvement in planned or actual warfarin use was 2.46 (95% CI 1.63-3.74). In addition, clinicians adhered to guidelines for antithrombotic therapy in patients with atrial fibrillation more frequently in the intervention group (control 70.4% vs. intervention 88.2%; p < 0.01). A program designed to identify hospitalized patients with non-valvular atrial fibrillation, assess their need for stroke prophylaxis, and initiate appropriate antithrombotic therapy led to an increase in planned antithrombotic, and most importantly, warfarin use upon discharge from the hospital. Confirmation that an increase in planned antithrombotic use upon discharge results in an actual increase in use after discharge is needed to determine the true effectiveness of this intervention.


Physical Dosimetric Reconstruction of a Radiological Accident at Nanjing (China) for Clinical Treatment Using Thudose.
A severe radiological accident involving an industrial radiography source containing Ir occurred in China. A worker was seriously exposed, which resulted in acute radiation syndrome. Initial whole-body dose was estimated at 1.51 Gy (95% Confidence Interval: 1.40-1.61 Gy) using biological dosimetry. This work performed a physical dosimetric reconstruction to provide more detailed exposure information for clinical treatment, using sitting and standing posture phantoms constructed by adjusting the Chinese reference adult male polygon surface phantoms to the worker body. A 3D view of photon flux in the body and dose distribution of local tissue with isodose lines in his legs were displayed by THUDose, and the absorbed doses of organs were present. These results were compatible with clinical symptoms and analysis, and they were helpful in assisting in the planning of therapy and in alerting physicians of potential high-risk organs. The physical dosimetric reconstruction could provide more detailed information for clinical treatment in a radiological accident with respect to obtaining local dose estimates.


Eco-efficient approaches to land management: a case for increased integration of crop and animal production systems.
Eco-efficiency is concerned with the efficient and sustainable use of resources in farm production and land management. It can be increased either by altering the management of individual crop and livestock enterprises or by altering the land-use system. This paper concentrates on the effects of crop sequence and rotation on soil fertility and nutrient use efficiency. The potential importance of mixed farming involving both crops and livestock is stressed, particularly when the systems incorporate biological nitrogen fixation and manure recycling. There is, however, little evidence that the trend in developed countries to farm-level specialization is being reduced. In some circumstances legislation to restrict diffuse pollution may provide incentives for more diverse eco-efficient farming and in other circumstances price premia for produce from eco-efficient systems, such as organic farming, and subsidies for the provision of environmental services may provide economic incentives for the adoption of such systems. However, change is likely to be most rapid where the present systems lead to obvious reductions in the productive potential of the land, such as in areas experiencing salinization. In other situations, there is promise that eco-efficiency could be increased on an area-wide basis by the establishment of linkages between farms of contrasting type, particularly between specialist crop and livestock farms, with contracts for the transfer of manures and, to a lesser extent, feeds.


Forensic autopsies from 1984 to 1993 in Vienna, Austria.
In Austria every death is subject to an examination by a medical doctor authorized by the local health authority. If death is suspected to be natural and/or perpetrated by another person, this doctor has to report it to the police. Depending on the investigation results, the examining magistrate in charge demands a judicial autopsy at the Institute of Forensic Medicine. In 1989, 41 murders of old patients by nursing assistants in a Viennese public hospital were disclosed. The main aim of this retrospective study was to determine any change in the demand for forensic autopsies by the Viennese health authority, as well as by the criminal court, after 1989. Furthermore, it was of interest to analyze the reporting practices of medical doctors examining corpses, as well as thr reaction of the criminal court during the study period. After 1989, there was a significant increase of non-judicial and judicial autopsies, performed by Viennese forensic pathologists. In addition, there was a significant increase of reports to the police by coroners as well as by forensic pathologists, paralleled by a higher rate of forensic autopsies demanded by the examining magistrate. This increase of forensic autopsies took place even though the overall rate of deaths in Vienna significantly decreased during the 10-year study period. Thus, the disclosure of 41 murders in the Viennese hospital in 1989 can be assumed as a turning point in the reporting practices of Viennese coroners, as well as the autopsy rate handled by Viennese forensic pathologists.


Early harvest and late transplantation as an effective therapeutic strategy in multiple myeloma.
Transplantation after high-dose chemotherapy prolongs survival in patients with multiple myeloma compared with standard therapy. It is unclear whether the optimal timing of transplantation is immediately after induction chemotherapy or whether stem cells may be cryopreserved for transplantation at subsequent progression or relapse. In this study, stem cells were collected within 6 months of diagnosis, followed by transplantation only at progression of myeloma. One hundred and eighteen patients with multiple myeloma had stem cells collected and cryopreserved. Eleven had transplants early in the disease after they demonstrated failure to respond to primary therapy. The remaining 107 were eligible for transplants when there was evidence of progressive disease. Of the 118 patients, 67 had transplants, nine died of progressive disease before transplantation, and 42 remain alive in plateau phase. The median survival of the group is 58.5 months; 67 are alive. Serum beta2-microglobulin, bone marrow labeling index (S phase), and hemoglobin level predicted overall survival (P < 0.006, P < 0.001, and P < 0.01, respectively). We conclude that early cryopreservation of blood stem cells followed by transplantation at progression is a feasible approach to therapy in patients with myeloma. The underlying biology of the disease has a greater impact on survival than the timing of transplantation. A prospective randomized trial is required to answer definitively the question of the optimal timing of blood cell transplantation.


The renal effects of sodium chlorite in the drinking water of C57L/J male mice.
The increasing concern over the possibility of adverse health effects due to the chlorination of drinking water has led to a search for alternative methods of disinfection. One potential alternative is chlorine dioxide which, when used, gives rise to the end product chlorite. This study investigated the potential of chlorite as a nephrotoxic agent when administered for various lengths of time (30, 90, and 180 days) in a series of concentrations (4, 20, and 100 ppm as sodium chlorite) to C57L/J male via drinking water. Examination by light and transmission electron microscopes did not reveal any evidence of renal pathologies. In addition, no evidence of significant mortality, water consumption differences, body weight change differences, kidney weight change differences, or differences in the percent kidney to body weight ratio was observed between the various levels of treatments.


Dopaminergic and clinical correlates of pathological gambling in Parkinson's disease: a case report.
Dopaminergic medication for motor symptoms in Parkinson's disease (PD) recently has been linked with impulse control disorders, including pathological gambling (PG), which affects up to 8% of patients. PG often is considered a behavioral addiction associated with disinhibition, risky decision-making, and altered striatal dopaminergic neurotransmission. Using [(11)C]raclopride with positron emission tomography, we assessed dopaminergic neurotransmission during Iowa Gambling Task performance. Here we present data from a single patient with PD and concomitant PG. We noted a marked decrease in [(11)C]raclopride binding in the left ventral striatum upon gambling, indicating a gambling-induced dopamine release. The results imply that PG in PD is associated with a high dose of dopaminergic medication, pronounced motor symptomatology, young age at disease onset, high propensity for sensation seeking, and risky decision-making. Overall, the findings are consistent with the hypothesis of medication-related PG in PD and underscore the importance of taking clinical variables, such as age and personality, into account when patients with PD are medicated, to reduce the risk of PG.


Cell type-specific angiotensin II-evoked signal transduction pathways: critical roles of Gbetagamma subunit, Src family, and Ras in cardiac fibroblasts.
Angiotensin II (Ang II) induces hypertrophy of cardiac myocytes and hyperplasia of cardiac fibroblasts. To determine the molecular mechanism by which Ang II displayed different effects on cardiac myocytes and fibroblasts, we examined signal transduction pathways leading to activation of extracellular signal-regulated kinases (ERKs). Ang II-induced ERK activation was abolished by pretreatment with pertussis toxin and by overexpression of the Gbetagamma subunit-binding domain of the beta-adrenergic receptor kinase 1 in cardiac fibroblasts but not in cardiac myocytes. Inhibition of protein kinase C strongly inhibited activation of ERKs by Ang II in cardiac myocytes, whereas inhibitors of tyrosine kinases but not of protein kinase C abolished Ang II-induced ERK activation in cardiac fibroblasts. Overexpression of C-terminal Src kinase (Csk), which inactivates Src family tyrosine kinases, suppressed the activation of transfected ERK in cardiac fibroblasts. Ang II rapidly induced phosphorylation of Shc and association of Shc with Grb2. Cotransfection of the dominant-negative mutant of Ras or Raf-1 kinase abolished Ang II-induced ERK activation in cardiac fibroblasts. Overexpression of Csk or the dominant-negative mutant of Ras had no effects on Ang II-induced ERK activation in cardiac myocytes. These findings suggest that Ang II-evoked signal transduction pathways differ among cell types. In cardiac fibroblasts, Ang II activates ERKs through a pathway including the Gbetagamma subunit of Gi protein, tyrosine kinases including Src family tyrosine kinases, Shc, Grb2, Ras, and Raf-1 kinase, whereas Gq and protein kinase C are important in cardiac myocytes.


Comparison of the efficiency of anti-androgenic regimens consisting of spironolactone, Diane 35, and cyproterone acetate in hirsutism.
The aim of the present study was to evaluate the effects of three different anti-androgenic drug-therapy regimens, Diane 35 (cyproterone acetate (CPA) [2 mg] and ethinyl estradiol [35 microg]) plus CPA, Diane 35 plus spironolactone, and spironolactone alone, in patients with hirsutism. In this prospective, randomized clinical study, 79 subjects with idiopathic hirsutismus were studied. The patients were divided into 3 groups. Group I patients (n=32) were treated with Diane 35 plus CPA, group II patients (n=25) with Diane 35 plus spironolactone [100 mg], and group III patients (n=22) with spironolactone [100 mg] alone. Serum FSH, LH, testosterone (T), and DHEAS levels were analyzed before and after treatment at 6 and 12 months. Hirsutism scores were graded according to the Ferriman-Gallwey scoring system, and side effects were monitored. All treatment regimens were found to be efficient and well-tolerated, and none of the patients stopped therapy due to any adverse event. However, in hormone screening, only patients on the Diane 35 plus CPA regimen revealed a decrease in serum T levels after therapy. As such, treatment of each hirsute patient should be planned individually, but with regard to both cost-efficiency and potential side effects, we recommend spironolactone alone in the treatment of hirsutismus.


Rapid quantification of myocardial fibrosis: a new macro-based automated analysis.
Fibrosis is associated with various cardiac pathologies and dysfunction. Current quantification methods are time-consuming and laborious. We describe a semi-automated quantification technique for myocardial fibrosis and validated this using traditional methods. Pulmonary Hypertension (PH) was induced in adult Wistar rats by subcutaneous monocrotaline (MCT) injection(40 mg/kg). Cryosections of myocardial tissue (5 μm) of PH rats (n = 9) and controls (n = 9) were stained using Picrosirius red and scanned with a digital microscopic MIRAX slide scanner. From these sections 21 images were taken randomly of each heart. Using ImageJ software a macro for automated image analysis of the amount of fibrosis was developed. For comparison, fibrosis was quantified using traditional polarisation microscopy. Both methods were correlated and validated against stereology as the gold standard. Furthermore, the method was tested in paraffin-embedded human tissues. Automated analysis showed a significant increase of fibrosis in PH hearts vs. control. Automated analysis correlated with traditional polarisation and stereology analysis (r(2) = 0.92 and r(2) = 0.95 respectively). In human heart, lungs, kidney, and liver, a similar correlation with stereology (r(2) = 0.91) was observed. Time required for automated analysis was 22% and 33% of the time needed for stereology and polarisation analysis respectively. Automated quantification of fibrosis is feasible, objective, and time-efficient.


Treatment outcomes of regenerative endodontic procedures in nonvital mature permanent teeth: a retrospective study.
This retrospective study was undertaken to clinically and radiographically evaluate the long-term outcomes of regenerative endodontic procedures (REPs) for nonvital mature permanent teeth, to analyze predictors influencing treatment outcomes. Nonvital mature permanent teeth treated by REPs with a minimum follow-up period of 6 months were included from 2015 to 2017. Treatment outcomes were categorized as success and failure. The periapical status and lesion healing were assessed in terms of the periapical index (PAI) and the percentage changes in periapical radiolucency (PARL) area. The clinical and radiographic outcomes of REPs were assessed by Mann-Whitney test at different follow-up period. Kaplan-Meier curves and Univariate Cox regression analysis were conducted to assess the success and identify potential predictors affecting outcomes, respectively. A total of 37 mature teeth with an average follow-up of 4.3 years satisfied the criteria, and 89.2% of the teeth had a successful outcome. Significant differences in PAI scores were found between each period with respect to the baseline (p < .05). Among different periods, there was a significant difference between intervals of 3-6 months and 7-12 months (p = .039) and no significant difference between each interval of more than 12 months (p > .05). Eighty-seven percent of teeth with preoperative PARL presented completely healed. REPs significantly decreased the PARL area at the interval of 7-12 months compared to 3-6 months (p = .025), with no significant difference between each interval of more than 12 months (p > .05). No significant predictor was found for the success of outcome (p > .05). Thirteen teeth (35.1%) regained pulp sensibility, and 40.5% of the teeth exhibited intracanal calcification. Within the limitations of this study, REPs provided a high long-term success rate and promoted the resolution of PARL as a biologically-based alternative treatment option for nonvital mature teeth. REPs provide a high long-term success rate and promoted healing of apical periodontitis comparable with reported outcomes for root canal therapy of mature teeth.


Surviving in isolation: genetic variation, bottlenecks and reproductive strategies in the Canarian endemic Limonium macrophyllum (Plumbaginaceae).
Oceanic archipelagos are typically rich in endemic taxa, because they offer ideal conditions for diversification and speciation in isolation. One of the most remarkable evolutionary radiations on the Canary Islands comprises the 16 species included in Limonium subsection Nobiles, all of which are subject to diverse threats, and legally protected. Since many of them are single-island endemics limited to one or a few populations, there exists a risk that a loss of genetic variation might limit their long-term survival. In this study, we used eight newly developed microsatellite markers to characterize the levels of genetic variation and inbreeding in L. macrophyllum, a species endemic to the North-east of Tenerife that belongs to Limonium subsection Nobiles. We detected generally low levels of genetic variation over all populations (H T = 0.363), and substantial differentiation among populations (F ST = 0.188; R ST = 0.186) coupled with a negligible degree of inbreeding (F = 0.042). Obligate outcrossing may have maintained L. macrophyllum relatively unaffected by inbreeding despite the species' limited dispersal ability and the genetic bottlenecks likely caused by a prolonged history of grazing. Although several factors still constitute a risk for the conservation of L. macrophyllum, the lack of inbreeding and the recent positive demographic trends observed in the populations of this species are factors that favour its future persistence.


Genome sequence of Desulfobacterium autotrophicum HRM2, a marine sulfate reducer oxidizing organic carbon completely to carbon dioxide.
Sulfate-reducing bacteria (SRB) belonging to the metabolically versatile Desulfobacteriaceae are abundant in marine sediments and contribute to the global carbon cycle by complete oxidation of organic compounds. Desulfobacterium autotrophicum HRM2 is the first member of this ecophysiologically important group with a now available genome sequence. With 5.6 megabasepairs (Mbp) the genome of Db. autotrophicum HRM2 is about 2 Mbp larger than the sequenced genomes of other sulfate reducers (SRB). A high number of genome plasticity elements (> 100 transposon-related genes), several regions of GC discontinuity and a high number of repetitive elements (132 paralogous genes Mbp(-1)) point to a different genome evolution when comparing with Desulfovibrio spp. The metabolic versatility of Db. autotrophicum HRM2 is reflected in the presence of genes for the degradation of a variety of organic compounds including long-chain fatty acids and for the Wood-Ljungdahl pathway, which enables the organism to completely oxidize acetyl-CoA to CO(2) but also to grow chemolithoautotrophically. The presence of more than 250 proteins of the sensory/regulatory protein families should enable Db. autotrophicum HRM2 to efficiently adapt to changing environmental conditions. Genes encoding periplasmic or cytoplasmic hydrogenases and formate dehydrogenases have been detected as well as genes for the transmembrane TpII-c(3), Hme and Rnf complexes. Genes for subunits A, B, C and D as well as for the proposed novel subunits L and F of the heterodisulfide reductases are present. This enzyme is involved in energy conservation in methanoarchaea and it is speculated that it exhibits a similar function in the process of dissimilatory sulfate reduction in Db. autotrophicum HRM2.


[Drosophila tumor suppressor Merlin is essential for morphogenesis of mitochondria during sperm formation].
Drosophila Merlin, an ortholog of the merlin protein encoded by the human Neurofibromatosis 2 (NF2) gene, is important for the regulation of cell proliferation and differentiation in the eye and wing. Also, it has been shown to be involved in male fertility in flies. In the present study, we formation using the comparative light and electron microscopic research of merlin mutants (mer3 and mer4) and ectopic expression of the Mer+ construct. Our work defines specific functions for Merlin in the mitochondria association and aggregation during the nebenkern formation and unfurling mitochondrial derivates during spermatid elongation. Possible role of Merlin as an adaptor protein that can link mitochondria with cytoskeleton is discussed.


The patient's differential diagnosis. Unpredictable concerns in visits for acute cough.
Agreement between physician and patient on the nature of the patient's illness problem is an important goal in primary care consultations. Unfortunately, both patient and doctor are often uncertain about the cause of the problem. The patient's satisfaction with a visit may be correlated with his or her agreement with the physician's differential diagnosis. Patients' diagnostic concerns elicited by interview just prior to and during 17 visits to family physicians for cough were compared with the doctors' differential diagnoses expressed during and after the visits using qualitative methods. Patients expressed a mean of 6.5 possibilities, of which a mean of 2.8 were also among the doctors' differential diagnoses. While many concerns were related to widely recognized popular ideas about disease, some patients had idiosyncratic, unpredictable diagnostic concerns about serious illness based on experiences in their families. Concerns were often expressed very indirectly during the visits. The physician's exploration of patients' uncertainty about the nature of their illnesses may help to elicit unexpected concerns that might otherwise prevent agreement on the nature of the problems.


Crystal structures of apo- and holo-bovine alpha-lactalbumin at 2. 2-A resolution reveal an effect of calcium on inter-lobe interactions.
High affinity binding of Ca(2+) to alpha-lactalbumin (LA) stabilizes the native structure and is required for the efficient generation of native protein with correct disulfide bonds from the reduced denatured state. A progressive increase in affinity of LA conformers for Ca(2+) as they develop increasingly native structures can account for the tendency of the apo form to assume a molten globule state and the large acceleration of folding by Ca(2+). To investigate the effect of calcium on structure of bovine LA, x-ray structures have been determined for crystals of the apo and holo forms at 2.2-A resolution. In both crystal forms, which were grown at high ionic strength, the protein is in a similar global native conformation consisting of alpha-helical and beta-subdomains separated by a cleft. Even though alternative cations and Ca(2+) liganding solvent molecules are absent, removal of Ca(2+) has only minor effects on the structure of the metal-binding site and a structural change was observed in the cleft on the opposite face of the molecule adjoining Tyr(103) of the helical lobe and Gln(54) of the beta-lobe. Changes include increased separation of the lobes, loss of a buried solvent molecule near the Ca(2+)-binding site, and the replacement of inter- and intra-lobe H-bonds of Tyr(103) by interactions with new immobilized water molecules. The more open cleft structure in the apo protein appears to be an effect of calcium binding transmitted via a change in orientation of helix H3 relative to the beta-lobe to the inter-lobe interface. Calcium is well known to promote the folding of LA. The results from the comparison of apo and holo structures of LA provide high resolution structural evidence that the acceleration of folding by Ca(2+) is mediated by an effect on interactions between the two subdomains.


Coping by stroke caregivers: sex similarities and differences.
Informal care provision presents many challenges. As the population ages, more people are forced to enter the role of informal caretaker. Despite the increase in the need for caregivers and the importance of providing care, there is little empirical research examining how men and women approach and cope with providing care. The current study examined stroke patients and their care providers to assess possible gender differences in the impact of caretaking on caretakers and care recipients. Results indicate no significant difference in patient well-being based on the gender of the caregiver. Some measures indicated that men have advantages as caregivers. Results are discussed with regard to the accuracy and utility of sex-role stereotypes about caregivers. In addition, these data provide potential insight about how to decrease caregiver burden, delay long-term hospitalization of the patient, and increase the quality of life for caretakers and patients.


Effects of Adaptation on Discrimination of Whisker Deflection Velocity and Angular Direction in a Model of the Barrel Cortex.
Two important stimulus features represented within the rodent barrel cortex are velocity and angular direction of whisker deflection. Each cortical barrel receives information from thalamocortical (TC) cells that relay information from a single whisker, and TC input is decoded by barrel regular-spiking (RS) cells through a feedforward inhibitory architecture (with inhibition delivered by cortical fast-spiking or FS cells). TC cells encode deflection velocity through population synchrony, while deflection direction is encoded through the distribution of spike counts across the TC population. Barrel RS cells encode both deflection direction and velocity with spike rate, and are divided into functional domains by direction preference. Following repetitive whisker stimulation, system adaptation causes a weakening of synaptic inputs to RS cells and diminishes RS cell spike responses, though evidence suggests that stimulus discrimination may improve following adaptation. In this work, I construct a model of the TC, FS, and RS cells comprising a single barrel system-the model incorporates realistic synaptic connectivity and dynamics and simulates both angular direction (through the spatial pattern of TC activation) and velocity (through synchrony of the TC population spikes) of a deflection of the primary whisker, and I use the model to examine direction and velocity selectivity of barrel RS cells before and after adaptation. I find that velocity and direction selectivity of individual RS cells (measured over multiple trials) sharpens following adaptation, but stimulus discrimination using a simple linear classifier by the RS population response during a single trial (a more biologically meaningful measure than single cell discrimination over multiple trials) exhibits strikingly different behavior-velocity discrimination is similar both before and after adaptation, while direction classification improves substantially following adaptation. This is the first model, to my knowledge, that simulates both whisker deflection velocity and angular direction and examines the ability of the RS population response to pinpoint both stimulus features within the context of adaptation.


Near-infrared fluorescent probes based on piperazine-functionalized BODIPY dyes for sensitive detection of lysosomal pH.
Three acidotropic, near-infrared fluorescent probes based on piperazine-modified BODIPY dyes (A, B and C) have been developed for the sensitive and selective detection of lysosomal pH in living cells. Probes A and B display low solubilities in aqueous solutions, whereas probe C is highly water-soluble. The fluorescent responsive mechanism of these probes to lysosomal pH is based on intramolecular charge transfer (ICT) and potential photo-induced electron transfer from piperazine moieties at 3,5-positions to BODIPY cores in the near-infrared region. The sensitivity and selectivity of the probes to pH over metal ions have been investigated by spectroscopic analysis in aqueous solutions. The probes have low auto-fluorescence at physiological pH conditions, whereas their fluorescence intensities significantly increase when pH is shifted to an acidic condition. Furthermore, these three probes were successfully applied to the in vitro lysosome imaging inside normal endothelial and breast cancer cells.


Lack of association of the CD14 promoter polymorphism--159C/T with Caucasian inflammatory bowel disease.
The inflammatory bowel diseases (IBDs), including Crohn's disease (CD) and ulcerative colitis (UC), are multifactorial diseases resulting from a complex interaction of genetic and environmental factors. The recently described CARD15 and TNF-alpha risk alleles are believed to be contributors to disease by disrupting inflammatory pathways via impaired response to bacteria. Other bacterial receptors, such as CD14, may also have a role in disease. A promoter polymorphism (-159C/T) in CD14 has been implicated in IBD in a number of studies. We have analysed this CD14 promoter polymorphism in probands from 206 multiplex IBD families, 110 sporadic IBD individuals and 189 healthy controls from the Australian population, all of whom are Caucasian. We could not replicate the described association between the CD14-159T allele and CD or UC, nor did we find any evidence for an interaction between the CARD15 or TNF-alpha risk alleles and the CD14-159T allele. It is possible that the association seen in other studies may be due to population stratification or to the CD14 polymorphism being in linkage with the real disease-causing variant(s).


Growth hormone secretion in primary adrenal Cushing's syndrome is disorderly and inversely correlated with body mass index.
To evaluate the impact on the somatotropic axis of endogenous cortisol excess in the absence of primary pituitary disease, we investigated spontaneous 24-h growth hormone (GH) secretion in 12 adult patients with ACTH-independent hypercortisolism. Plasma GH concentration profiles (10-min samples) were analyzed by deconvolution to reconstruct secretion and approximate entropy to quantitate orderliness of the release process. Comparisons were made with a body mass index (BMI)-, age-, and gender-matched control group and an age- and gender-matched lean control group. GH secretion rates did not differ from BMI-matched controls but were twofold lower compared with lean subjects, mainly due to a 2.5-fold attenuation of the mean secretory burst mass (P = 0.001). In hypercortisolemic patients, GH secretion was negatively correlated with BMI (R = -0.55, P = 0.005) but not cortisol secretion. Total serum IGF-I concentrations were similar in the three groups. Approximate entropy (ApEn) was increased in patients with Cushing's syndrome compared with both control groups (vs. BMI-matched, P = 0.04; vs. lean, P = 0.001), denoting more irregular GH secretion patterns. ApEn in patients correlated directly with cortisol secretion (R = 0.77, P = 0.003). Synchrony between cortisol and GH concentration series was analyzed by cross-correlation, cross-ApEn, and copulsatility analyses. Patients showed loss of pattern synchrony compared with BMI-matched controls, but copulsatility was unchanged. We conclude that hyposomatotropism in primary adrenal hypercortisolism is only partly explained (approximately 30%) by increased body weight and that increased GH secretory irregularity and loss of synchrony suggest altered coordinate regulation of GH release.