Parthenogenetic activation of mouse eggs by microinjection of a truncated c-kit tyrosine kinase present in spermatozoa.
A truncated form of the c-kit tyrosine kinase receptor, corresponding to the phosphotransferase portion of the cytoplasmic catalytic domain and the carboxyterminus (tr-kit), is accumulated during late mouse spermiogenesis. Here we report that tr-kit is specifically localized in the residual sperm cytoplasm, with maximal accumulation in the midpiece of the flagellum, suggesting that it can enter the egg during fertilization. Microinjection of extracts from COS cells expressing a recombinant tr-kit protein into metaphase II-arrested mouse oocytes caused complete oocyte activation, including cortical granule exocytosis, completion of the 2nd meiotic division, formation of a parthenogenetic pronucleus and progression through cleavage stages. No activation above background levels was obtained with extracts from mock-transfected COS cells. Similar results were obtained by microinjection of in vitro synthesized tr-kit mRNA into metaphase II-arrested oocytes. Tr-kit-induced parthenogenetic egg activation was completely inhibited by oocyte preincubation with the Ca2(+)-chelating agent BAPTA-AM or with a specific inhibitor of phospholipase C activity. Tr-kit-induced egg activation was associated with a decrease in activity of mitogen-activated protein kinase, an essential component of the cytostatic factor. These results candidate tr-kit as a putative sperm factor required for triggering activation of mouse eggs at fertilization.


From political to personal violence: Links between conflict and non-partner physical violence in post-conflict Liberia.
War and interpersonal violence together account for a large burden on global health. Yet very few studies look at the relationship between these types of aggression. Non-partner physical violence (NPPV) is an often-understudied form of gender-based violence (GBV). This analysis draws on two datasets from one conflict-affected country, Liberia, to evaluate the impact of conflict on NPPV post-conflict. The Armed Conflict Location and Event Dataset (ACLED) measures the intensity of the conflict in Liberia from 1999-2003, while the Demographic and Heath Survey (DHS) data measure women's experiences with violence four years post-conflict. Almost half of women surveyed (45%) indicated that they experienced any kind of NPPV, highlighting the widespread nature of this issue. A multilevel modelling approach was used to account for the nesting of individuals within districts. Women living in districts that experienced conflict events in four or five years were almost three times as likely (aOR 2.93, p < .001) to experience past-year NPPV compared to individuals living in no conflict districts. Findings from this study suggest women residing in a conflict event-affected district may be at heightened risk of increased violence even years after peace is declared.


Towards a reconceptualization of striatal interactions between glutamatergic and dopaminergic neurotransmission and their contribution to the production of movements.
According to the current model of the basal ganglia organization, simultaneous activation of the striato-nigral direct pathway by glutamatergic and dopaminergic neurotransmission should lead to a synergistic facilitatory action on locomotor activity, while in contrast activation of the indirect pathway by these two neurotransmittions should lead to antagonistic effects on locomotor activity. Based on published data, as a break with the current thinking, we propose a reconceptualization of functional interactions between dopaminergic and glutamatergic neurotransmission. In this model, dopaminergic neurotransmission is seen as a motor pacemaker responsible for the basal and primary activation of striatal output neurons and glutamate as a driver providing a multiple combination of tonic, phasic, facilitatory and inhibitory influxes resulting from the processing of environmental, emotional and mnesic stimuli. Thus, in the model, glutamate-coded inputs would allow tuning the intrinsic motor-activating properties of dopamine to adjust the production of locomotor activity into goal-oriented movements.


Identification and characterization of a calmodulin-binding domain in Ral-A, a Ras-related GTP-binding protein purified from human erythrocyte membrane.
A 28-kDa protein (p28) has been purified from Triton X-100 extracts of human erythrocyte plasma membrane by calmodulin affinity chromatography. Based on internal peptide sequencing and its protein amino acid composition, this protein has been shown to be highly related, if not identical to, Ral-A, a Ras-related GTP-binding protein. This protein assignment is consistent with the findings that p28 binds [32P]GTP specifically and has low GTPase activity. In this study we describe the identification and characterization of a calmodulin-binding domain in Ral-A. The Ca2+-dependent interaction of p28 with calmodulin was first detected by a calmodulin affinity column. Gel overlay experiments of both p28 and recombinant Ral-A with biotinylated calmodulin provided strong evidence that Ral-A is a calmodulin-binding protein. A peptide of 18 residues (P18) with the sequence SKEKNGKKKRKSLAKRIR has been identified as a putative calmodulin-binding domain in Ral-A, because it comprises a basic/hydrophobic composition with the propensity to form an amphiphilic helix. P18 was synthesized, and its interaction with calmodulin by gel overlay was shown to be Ca2+-dependent. Circular dichroism analysis demonstrated that this interaction results in less alpha-helical content upon calmodulin complex formation. These results indicate that Ral-A is a calmodulin-binding protein, raising the possibility that it may be associated with Ca2+-dependent intracellular signaling pathways.


Cytotoxic T lymphocyte responses against alloantigens exhibit preferential effector cell activity for H-2K or H-2D region products similar to that for H-2 restricted responses.
The present study was designed to investigate the patterns of allogeneic cytotoxic T cell responses against H-2K and H-2D region products. B10.D2 anti-B10.A(2R) effector cells lysed K-end matched target cells approximately 5 times more efficiently than D-end matched target cells. In contrast, C57BL/10 anti-B10.A effector cells killed K-end or D-end matched target cells equally well, whereas C57BL/10 anti-B10.D2 CTL exhibited a slight preference for D-end matched target cells. These results demonstrate that whole H-2 disparate stimulating populations do not necessarily induce equivalent h-2K and H-2D region-specific CTL responses. The strength of the cytotoxic responses was found to correlate with the number of known serologically detectable specificities shared between the stimulating and target cell populations. The similarities between these allogeneic and MHC-restricted CTL response patterns (which are under Ir-like control) raise the possibility that this regulation reflects the number of K or D region self-antigens that can be recognized in association with a foreign antigenic determinant and suggests that allogeneic responses to major histocompatibility antigens are also self-restricted by MHC gene products. The possibility is considered that such Ir-like control is also a reflection of the amounts of H-2K and H-2D products shed from the cell surface.


Faunal Burrows Alter the Diversity, Abundance, and Structure of AOA, AOB, Anammox and n-Damo Communities in Coastal Mangrove Sediments.
In the present work, the diversity, community structures, and abundances of aerobic ammonia-oxidizing archaea (AOA) and bacteria (AOB), anaerobic ammonium-oxidizing (anammox) bacteria, and denitrifying anaerobic methane oxidization (n-damo) bacteria were unraveled in the bioturbated areas of the coastal Mai Po mangrove sediments. Results indicated that the bioturbation by burrowing in mangrove sediments was associated with higher concentration of NH4+ but lower concentrations of both NO2- and NO3-, and increase in diversity and richness of both AOA and AOB, but relatively lower diversity and richness of n-damo bacteria. The phylotypes of anammox bacterial community were significantly increased while their phylogenetic lineages observed in the less bioturbated areas were also maintained. Infauna also showed a great impact on the composition of n-damo bacterial phylotypes and burrowing activity altered the n-damo community structure profoundly in the sampled areas. The communities of n-damo bacteria in the surrounding bulk sediments showed similar structures to marine n-damo communities, but those on the burrow wall and in the ambient surface layer had a freshwater pattern, which was different from previous findings in Mai Po wetland. On the other hand, the abundances of AOA, AOB, and n-damo bacteria were greatly stimulated on burrow walls while the abundance of anammox bacteria remained unchanged. Infaunal burrows and mangrove roots affected the relative abundance of AOA and AOB. The benthic infauna stimulated the abundances of AOA, AOB, anammox, and n-damo bacteria. Furthermore, NH4+ and NO2- were important environmental factors changing the structure of each group. The communities of anammox and n-damo bacteria in bioturbated areas showed a competitive relationship.


Emulsion-Based Intradermal Delivery of Melittin in Rats.
Bee venom (BV) has long been used as a traditional medicine. The aim of the present study was to formulate a BV emulsion with good rheological properties for dermal application and investigate the effect of formulation on the permeation of melittin through dermatomed rat skin. A formulated emulsion containing 1% (w/v) BV was prepared. The emulsion was compared with distilled water (DW) and 25% (w/v) N-methyl-2-pyrrolidone (NMP) in DW. Permeation of melittin from aqueous solution through the dermatomed murine skin was evaluated using the Franz diffusion cells. Samples of receptor cells withdrawn at pre-determined time intervals were measured for melittin amount. After the permeation study, the same skin was used for melittin extraction. In addition, a known amount of melittin (5 μg/mL) was added to stratum corneum, epidermis, and dermis of the rat skin, and the amount of melittin was measured at pre-determined time points. The measurement of melittin from all samples was done with HPLC-MS/MS. No melittin was detected in the receptor phase at all time points in emulsion, DW, or NMP groups. When the amount of melittin was further analyzed in stratum corneum, epidermis, and dermis from the permeation study, melittin was still not detected. In an additional experiment, the amount of melittin added to all skin matrices was corrected against the amount of melittin recovered. While the total amount of melittin was retained in the stratum corneum, less than 10% of melittin remained in epidermis and dermis within 15 and 30 min, respectively. Skin microporation with BV emulsion facilitates the penetration of melittin across the stratum corneum into epidermis and dermis, where emulsified melittin could have been metabolized by locally-occurring enzymes.


Supraorbital neuralgia.
Supraorbital neuralgia is a rare disorder clinically characterized by the following triad: 1) forehead pain in the territory supplied by the supraorbital nerve, without side shift; 2) tenderness on either the supraorbital notch or traject of the nerve; and 3) absolute, but transitory relief of symptoms upon supraorbital nerve blockade. The pain presents with a chronic or intermittent pattern. In addition, there may be signs and symptoms of sensory dysfunction (hypoesthesia, paresthesia and allodynia), and typical "neuralgic features" (lightning pain and exteroceptive precipitating mechanisms). However, sensitive and neuralgic features are not constantly present and seem to be more frequent in the secondary, usually post-traumatic, forms.


The molecular mechanism of the effects of the anti-neuropathic ligands on the modulation of the Sigma-2 receptor: An in-silico study.
Neuropathic pain (NP) is a prevalent medical condition that lacks an effective treatment. Recently, the Sigma-2 receptor (S2R) has been proposed as a potential therapeutic target for NP. Some highly-selective S2R ligands (UKH1114, CM398, and YTD) have shown promising results in vivo, but the molecular interaction between the S2R and these ligands has been scarcely investigated. This work explores changes in the S2R upon interaction with the three mentioned ligands using in silico approaches. The results indicated that the ICL1, H1, ICL2, and ECL are the most dynamic regions of S2R in all systems. Binding interaction analysis identified amino acids with significant contribution to the binding free energy. Notably, the UKH1114-S2R simulation trajectory revealed that small alterations in the ICL1, H1, ICL2, and ECL form a new stable opening in the S2R, linking the occluded S2R binding pocket to the endoplasmic reticulum lumen, providing more evidence for the assumptions about the EBP and S2R mechanism of function. Further, the agreement between the membrane parameters in our study and experimental values confirms the validity of the MD simulations. Overall, this study provides new insights into the interaction between anti-NP ligands and the S2R.


Evaluation of pre- and posttransplant donor-specific transfusion/cyclosporine A in non-HLA identical living donor kidney transplant recipients. Cooperative Clinical Trials in Transplantation Research Group.
The beneficial effects of donor specific transfusion (DST) have become controversial in the cyclosporine era. This study was performed to evaluate the potential benefits of a new protocol for administering DSTs in the perioperative period. Non-HLA identical living donor kidney transplant recipients were randomized prospectively to control or to receive a DST 24 hr before transplant and 7-10 days posttransplant. All patients received similar immunosuppression according to protocol. The protocol had 212 evaluable patients (115 transfused and 97 control). There were no differences in 1- and 2-year graft and patient survival, causes of graft failure, incidence and types of infection, repeat hospitalization, or the ability to withdraw steroids. Immunological hyporesponsiveness (by mixed lymphocyte culture) occurred more frequently in transfused patients (18%) than controls (3%) (P = 0.04). Blood stored for > or =3 days was associated with fewer early rejections than blood stored < or =2 days. Overall, class II antigen mismatches were associated with more rejection episodes than class I antigen mismatches. However, transfused patients, but not control patients, with more class I antigen mismatches were more likely to have rejections. Administration of DSTs by the method described had no practical influence on patient or graft survival for up to 2 years. However, donor-specific hyporesponsiveness was more common in transfused patients (18 vs. 3%). Longer follow-up will be needed to determine whether DST will be associated with long-term benefit.


Combined quality function deployment and logical framework analysis to improve quality of emergency care in Malta.
The purpose of this paper is to develop an integrated patient-focused analytical framework to improve quality of care in accident and emergency (A & E) unit of a Maltese hospital. The study adopts a case study approach. First, a thorough literature review has been undertaken to study the various methods of healthcare quality management. Second, a healthcare quality management framework is developed using combined quality function deployment (QFD) and logical framework approach (LFA). Third, the proposed framework is applied to a Maltese hospital to demonstrate its effectiveness. The proposed framework has six steps, commencing with identifying patients' requirements and concluding with implementing improvement projects. All the steps have been undertaken with the involvement of the concerned stakeholders in the A & E unit of the hospital. The major and related problems being faced by the hospital under study were overcrowding at A & E and shortage of beds, respectively. The combined framework ensures better A & E services and patient flow. QFD identifies and analyses the issues and challenges of A & E and LFA helps develop project plans for healthcare quality improvement. The important outcomes of implementing the proposed quality improvement programme are fewer hospital admissions, faster patient flow, expert triage and shorter waiting times at the A & E unit. Increased emergency consultant cover and faster first significant medical encounter were required to start addressing the problems effectively. Overall, the combined QFD and LFA method is effective to address quality of care in A & E unit. PRACTICAL/IMPLICATIONS: The proposed framework can be easily integrated within any healthcare unit, as well as within entire healthcare systems, due to its flexible and user-friendly approach. It could be part of Six Sigma and other quality initiatives. Although QFD has been extensively deployed in healthcare setup to improve quality of care, very little has been researched on combining QFD and LFA in order to identify issues, prioritise them, derive improvement measures and implement improvement projects. Additionally, there is no research on QFD application in A & E. This paper bridges these gaps. Moreover, very little has been written on the Maltese health care system. Therefore, this study contributes demonstration of quality of emergency care in Malta.


A prospective study of patients with radiological pyelonephritis, papillary necrosis and obstructive atrophy.
Twenty-six patients with radiological unilateral chronic pyelonephritis, 36 patients with bilateral chronic pyelonephritis, 14 patients with papillary necrosis and nine patients with obstructive atrophy have been followed from five to 135 months for a total of 374 patient years. Serial changes in renal function and pyelographic appearances have been correlated with bacteriuria, analgesic ingestion, blood pressure and reflux. The calculated survival rate at five years was 95 per cent for patients with bilateral pyelonephritis and 92 per cent for patients with papillary necrosis. The ten-year survival rate was 86 per cent and 56 per cent respectively. The survival rate for patients with unilateral pyelonephritis and obstructive atrophy was 100 per cent at five and ten years. Bacteriuria was not associated with deteriorating renal function determined by serial plasma creatinine estimations. Although all patients in whom there was some radiographic change had bacteriuria on later review, other factors, including excess analgesic intake, reflux and stones were recognized in most. There was a high incidence of analgesic ingestion among patients whose renal function declined and in whom there was some change in serial radiographs. The prevalence of hypertension among patients with normal renal function was 12 and 28 per cent for patients with unilateral pyelonephritis and bilateral pyelonephritis respectively. There was a significant increase in both blood urea and plasma creatinine in all patients with hypertension (diastolic pressure greater than 90 mm Hg) and a much higher prevalence of hypertension in patients whose plasma creatinine exceeded 1.3 mg/100 ml. Thrity per cent of patients with unilateral pyelonephritis and 50 per cent with bilateral pyelonephritis had vesicoureteric reflux of varying degrees. There was no evidence to suggest that major degrees of reflux (grade 3) was associated with further renal damage. These observations indicate the benign course of the majority of patients with radiological pyelonephritis. Control of blood pressure, and analgesic intake will help to preserve renal function whilst prevention of symptomatic urinary infection by long term low dose therapy will reduce morbidity.


Site-specific incorporation of p-propargyloxyphenylalanine in a cell-free environment for direct protein-protein click conjugation.
The tyrosine analog p-propargyloxyphenylalanine (pPa), like tyrosine, has limited water solubility. It has been postulated that this limited solubility has contributed to reduced cellular uptake of pPa and thus reduced in vivo incorporation of pPa into proteins. Using a cell-free protein synthesis system (CFPS) to circumvent cellular uptake, pPa has been incorporated site-specifically into proteins with high specificity at yields up to 27 times greater than the highest previously reported yield. The alkyne group present on proteins incorporated with pPa provides a reactive residue for site-specific bioconjugation with the copper(I)-catalyzed azide-alkyne [3 + 2] cycloaddition (CuAAC). Previously, incorporation of another CuAAC-compatible unnatural amino acid p-azido-l-phenylalanine (pAz) was demonstrated with CFPS. However, incorporation of pPa may be preferred over pAz due to the instability of the pAz's aryl-azido moiety upon UV or near-UV light exposure. Also, the ability to incorporate site-specifically both reactants of the CuAAC (the alkyne group of pPa and the azido group of pAz) into proteins enables direct site-specific conjugation of heterologous proteins. We have demonstrated (for the first time to our knowledge) a one-step, linker-less, site-specific, direct protein-to-protein conjugation using CuAAC and unnatural amino acids.


The classification and diagnosis of erythrocytosis.
An absolute erythrocytosis is present when the red cell mass is raised and the haematocrit is elevated above prescribed limits. Causes of an absolute erythrocytosis can be primary where there is an intrinsic problem in the bone marrow and secondary where there an event outside the bone marrow driving erythropoiesis. This can further be divided into congenital and acquired causes. There remain an unexplained group idiopathic erythrocytosis. Investigation commencing with thorough history taking and examination and then investigation depending on initial features is required. Clear simple criteria for polycythaemia vera are now defined. Those who do not fulfil these criteria require further investigation depending on the clinical scenario and initial results. The erythropoietin level provides some guidance as to the direction in which to proceed and the order and extent of investigation necessary in an individual patient. It should thus be possible to make an accurate diagnosis in the majority of patients.


Molecular cloning and expression of human tumor necrosis factor and comparison with mouse tumor necrosis factor.
U-937 cells, a monocytic line derived from a human histiocytic lymphoma, were induced for human tumor necrosis factor (TNF) secretion into the medium and were used for the preparation of TNF mRNA. Biological activity of the latter was quantified in a Xenopus laevis oocyte injection system. TNF mRNA was enriched by gradient centrifugation and this size-fractionated mRNA was used for synthesis of cDNA and inserted into the unique PstI site of pAT153. A recombinant plasmid containing human TNF cDNA was selected by colony hybridization using an internal fragment of a mouse TNF cDNA clone [Fransen, L., Mueller, R., Marmenout, A., Tavernier, J., Van der Heyden, J., Kawashima, E., Chollet, A., Tizard, R., Van Heuverswyn, H., Van Vliet, A., Ruysschaert, M. R. & Fiers, W. (1985) Nucleic Acids Res. 13, 4417-4429] as a probe. The sequence of this human TNF cDNA is in agreement with the one published by Pennica et al. [Pennica, D., Nedwin, G. E., Hayflick, J. S., Seeburg, P. H., Derynck, R., Palladino, M. A., Kohr, W. J., Aggarwal, B. B. & Goeddel, D. V. (1984) Nature (Lond.) 312, 724-729]. The 157-amino-acid-long mature sequence is about 80% homologous to mouse TNF and its hydrophilicity plot is also very similar, in spite of the apparent species specificity of TNF. In contrast to mouse TNF, it contains no potential N-glycosylation site. When compared to other cytokines, like IFN-beta, IFN-gamma, or IL-2, there is a remarkably high preference for G X C pairs in the third-letter positions. Expression of the TNF cDNA in monkey COS cells or in Escherichia coli gives rise to a protein having similar biological and serological properties as natural human TNF. A human genomic clone was also identified and sequenced; it was found to be in good agreement with the one recently published by Shirai et al. [Shirai, T., Yamaguchi, H., Ito, H., Todd, C. W. & Wallace, R. B. (1985) Nature (Lond.) 313, 803-806], except for some differences in the introns and 5'-untranslated region.


SUN1 facilitates CHMP7 nuclear influx and injury cascades in sporadic amyotrophic lateral sclerosis.
We have recently identified the aberrant nuclear accumulation of the ESCRT-III protein CHMP7 as an initiating event that leads to a significant injury to the nuclear pore complex (NPC) characterized by the reduction of specific nucleoporins (Nups) from the neuronal NPC in sporadic ALS (sALS) and C9orf72 ALS/FTD induced pluripotent stem cell (iPSC) derived neurons (iPSNs), a phenomenon also observed in postmortem patient tissues. Importantly, this NPC injury is sufficient to contribute to TDP-43 dysfunction and mislocalization, a common pathological hallmark of neurodegenerative diseases. However, the molecular mechanisms and events that give rise to increased nuclear translocation and/or retention of CHMP7 to initiate this pathophysiological cascade remain largely unknown. Here, using an iPSN model of sALS, we demonstrate that impaired NPC permeability barrier integrity and interactions with the LINC complex protein SUN1 facilitate CHMP7 nuclear localization and the subsequent "activation" of NPC injury cascades. Collectively, our data provide mechanistic insights in the pathophysiological underpinnings of ALS/FTD and highlight SUN1 as a potent contributor to and modifier of CHMP7 mediated toxicity in sALS pathogenesis.


Two-phase learning-based 3D deblurring method for digital breast tomosynthesis images.
In digital breast tomosynthesis (DBT) systems, projection data are acquired from a limited number of angles. Consequently, the reconstructed images contain severe blurring artifacts that might heavily degrade the DBT image quality and cause difficulties in detecting lesions. In this study, we propose a two-phase learning approach for artifact compensation in a coarse-to-fine manner to mitigate blurring artifacts effectively along all viewing directions of the DBT image volume (i.e., along the axial, coronal, and sagittal planes) to improve the detection performance of lesions. The proposed method employs a convolutional neural network model comprising two submodels/phases, with Phase 1 performing three-dimensional (3D) deblurring and Phase 2 performing additional 2D deblurring. To investigate the effects of loss functions on the proposed model's deblurring performance, we evaluated several loss functions, such as the pixel-based loss function, adversarial-based loss function, and perception-based loss function. Compared with the DBT image, the mean squared error of the image and the root mean squared errors of the gradient of the image decreased by 82.8% and 44.9%, respectively, and the contrast-to-noise ratio increased by 183.4% in the in-focus plane. We verified that the proposed method sequentially restored the missing frequency components as the DBT images were processed through the Phase 1 and Phase 2 steps. These results indicate that the proposed method performs effective 3D deblurring, significantly reducing the blurring artifacts in the in-focus plane and other planes of the DBT image, thus improving the detection performance of lesions.


Coarse particulate matter associated with increased risk of emergency hospital admissions for pneumonia in Hong Kong.
Epidemiological research on the effects of coarse particles (PMc, particulate matter between 2.5 and 10 μm in aerodynamic diameter) on respiratory morbidity is sparse and inconclusive. Pneumonia is an inflammatory condition of lung caused by infections, which may be triggered and exacerbated by PMc exposure. To estimate the effect of PMc on emergency hospital admissions for pneumonia after controlling for PM(2.5) and gaseous pollutants. PMc concentrations were estimated by subtracting PM(2.5) from PM(10) measurements in each of the 10 air monitoring stations from January 2011 to December 2012 in Hong Kong and then citywide daily average concentrations of PMc were computed from the 10 stations. Generalised additive Poisson models were used to examine the relationship between PMc and daily emergency hospital admissions for pneumonia, adjusting for PM(2.5) and gaseous pollutants (NO(2), SO(2) and O(3)). Subgroup analyses by gender and age were also performed to identify the most susceptible subpopulations. PMc and PM(2.5) were significantly associated with emergency pneumonia hospitalisations. Every 10 μg/m(3) increment of PMc in the past 4 days (lag0-lag3) was associated with a 3.33% (95% CI 1.54% to 5.15%) increase in emergency hospitalisations for pneumonia. The effect estimates of PMc were robust to the adjustment of PM(2.5), NO(2) or SO(2), but attenuated on the inclusion of O(3) in the model. Women, children and older people might be more vulnerable to PMc exposure. Short-term PMc exposure is associated with emergency hospitalisations for pneumonia in Hong Kong. Air quality regulation specifically for PMc might be considered.


[Clinical-pathogenic role of the functional condition of vegetative nervous system in progressive the mixed cryoglobulinemia in patients with chronic hepatitis C].
The study of 304 patients with chronic hepatitis C is presented. It was shown that in patients with chronic hepatitis C the occurrence and increase of violations of a functional condition of vegetative nervous system it is combined with emergence and progressing of signs of the mixed cryoglobulinemia. Clinical demonstration of the HCV associated crioglobulinemia syndrome is accompanied by the most expressed vegetative dysfunction, the being characterized low capacity of variability of a rhythm of heart and development of a vegetative disbalance and sympathicotonia.


[Early postoperative occlusion].
Early postoperative occlusion is an occlusive syndrome which either occurs immediately after operation or is delayed for a few days. It may result from many causes, with the four most common being a mechanical obstacle, inflammatory bowel occlusion, vascular ileus, and metabolic ileus. Each of these causes results in specific clinical features, but circumstances at onset are the most important distinctive criteria. Reoperation, which is viewed with anxiety by both the patient and surgeon, is not always requisite, although in some instances an emergency surgical procedure is needed.


K+ modulates genetic competence and the stress regulon of Streptococcus mutans.
Potassium (K+) is the most abundant cation in dental plaque fluid. Previously, we reported the link between K+ transport via Trk2 in Streptococcus mutans and its two critical virulence attributes: acid tolerance and surface adhesion. Herein, we build further on the intimate link between K+ levels and S. mutans biology. High (>25 mM) versus low (≤5 mM) K+ concentrations in the growth medium affected conformational epitopes of cell surface-localized adhesin P1. At low K+, the expression of stress response elements gcrR and codY, cell-adhesion-associated genes such as spaP and metabolism-associated genes such as bglP was induced at stationary phase (P<0.05), suggesting that K+-mediated regulation is growth phase-dependent and stress-sensitive. Production of the newly discovered secretory protein encoded by SMU_63c was strongly dependent on the availability of K+ and growth phase. This protein is a newly discovered regulator of genetic competence and biofilm cell density. Thus, the influence of K+ on DNA transformation efficiency was also examined. Compared with 25 mM K+ concentration, the presence of low K+ reduced the transformation frequency by 100-fold. Genetic transformation was abolished in a strain lacking a Trk2 system under all K+ concentrations tested. Consistent with these findings, repression of competence-associated genes, comS and comX, was observed under low environmental K+ conditions and in the strain lacking Trk2. Taken together, these results highlight a pivotal role for environmental K+ as a regulatory cation that modulates stress responses and genetic transformation in S. mutans.


Independent component analysis: mining microarray data for fundamental human gene expression modules.
As public microarray repositories rapidly accumulate gene expression data, these resources contain increasingly valuable information about cellular processes in human biology. This presents a unique opportunity for intelligent data mining methods to extract information about the transcriptional modules underlying these biological processes. Modeling cellular gene expression as a combination of functional modules, we use independent component analysis (ICA) to derive 423 fundamental components of human biology from a 9395-array compendium of heterogeneous expression data. Annotation using the Gene Ontology (GO) suggests that while some of these components represent known biological modules, others may describe biology not well characterized by existing manually-curated ontologies. In order to understand the biological functions represented by these modules, we investigate the mechanism of the preclinical anti-cancer drug parthenolide (PTL) by analyzing the differential expression of our fundamental components. Our method correctly identifies known pathways and predicts that N-glycan biosynthesis and T-cell receptor signaling may contribute to PTL response. The fundamental gene modules we describe have the potential to provide pathway-level insight into new gene expression datasets.


Identifying the health conscious consumer.
Individuals who lead a "wellness-oriented" lifestyle are concerned with nutrition, fitness, stress, and their environment. They accept responsibility for their health and are excellent customers for health-related products and services. Those who lack a wellness orientation are identified as higher health risks and become candidates for health promotion program intervention. The authors report a new scale by which to measure the wellness-oriented lifestyle. Scale development procedures are detailed, followed by information from five studies that support its validity. The authors suggest ways health care marketers may use the Wellness Scale to segment and target potential customers and position their products and services.


Radiolocalization of xenografted human malignant melanoma by a monoclonal antibody (9.2.27) to a melanoma-associated antigen in nude mice.
A murine monoclonal antibody (9.2.27), directed to a Mr 250,000 glycoprotein-chondroitan sulfate proteoglycan complex, was radiolabeled with 125I and assessed for radiolocalization in tumor and normal tissues of normal and tumor-bearing nude mice. The 125I-9.2.27 localized in vivo preferentially in Mr 250,000 antigen-expressing human melanomas (FMX-Met, SESX) but not in low antigen-expressing tumors (LOX-L) xenografted in nude mice. The imaging index of tumor cells was positively correlated with the antigen density of the various melanoma cell lines as measured by flow cytometry. The nonspecific immunoglobulin RPC-5 of the same IgG2a subclass as 9.2.27 did not specifically localize to xenografts of melanoma. The total amount of 125I-9.2.27 accumulated in the tumor was directly correlated with tumor size. However, the specific radioactivity (cpm/g) in smaller tumors was higher than that in larger tumors. Nonspecific uptake and circulating antibody levels differed between normals and tumor-bearers. The organs of the reticuloendothelial system of normal mice accumulated more labeled antibody than did those of tumor bearers, and conversely, tumor bearers had higher levels of circulating labeled antibody in the blood than normals. The circulating labeled antibody in tumor bearers was still monomeric but had no detectable antigen-binding capacity.


Maternal undernutrition during late gestation-induced intrauterine growth restriction in the rat is associated with impaired placental GLUT3 expression, but does not correlate with endogenous corticosterone levels.
Fetal intrauterine growth restriction (IUGR) is a frequently occurring and serious complication of pregnancy. Infants exposed to IUGR are at risk for numerous perinatal morbidities, including hypoglycemia in the neonatal period, as well as increased risk of later physical and/or mental impairments, cardiovascular disease and non-insulin-dependent diabetes mellitus. Fetal growth restriction most often results from uteroplacental dysfunction during the later stage of pregnancy. As glucose, which is the most abundant nutrient crossing the placenta, fulfills a large portion of the fetal energy requirements during gestational development, and since impaired placental glucose transport is thought to result in growth restriction, we investigated the effects of maternal 50% food restriction (FR50) during the last week of gestation on rat placental expression of glucose transporters, GLUT1, GLUT3 and GLUT4, and on plasma glucose content in both maternal and fetal compartments. Moreover, as maternal FR50 induces fetal overexposure to glucocorticoids and since these hormones are potent regulators of placental glucose transporter expression, we investigated whether putative alterations in placental GLUT expression correlate with changes in maternal and/or fetal corticosterone levels. At term (day 21 of pregnancy), plasma glucose content was significantly reduced (P<0.05) in mothers subjected to FR50, but was not affected in fetuses. Food restriction reduced maternal body weight (P<0.001) but did not affect placental weight. Plasma corticosterone concentration, at term, was increased (P<0.05) in FR50 mothers. Fetuses from FR50 mothers showed reduced body weight (P<0.001) but higher plasma corticosterone levels (P<0.05). Adrenalectomy (ADX) followed by corticosterone supplementation of the mother prevented the FR50-induced rise in maternal plasma corticosterone at term. Food restriction performed on either sham-ADX or ADX mothers induced a similar reduction in the body weight of the pups at term (P<0.01). Moreover, plasma corticosterone levels were increased in pups from sham-ADX FR50 mothers (P<0.01) and in pups from ADX control mothers (P<0.01). Western blot analysis of placental GLUT proteins showed that maternal FR50 decreased placental GLUT3 protein levels in all experimental groups at term (P<0.05 and P<0.01), but did not affect either GLUT1 or GLUT4 protein levels. Northern blot analysis of placental GLUT expression showed that both GLUT1 and GLUT3 mRNA were not affected by the maternal feeding regimen or surgery. We concluded that prolonged maternal malnutrition during late gestation decreases maternal plasma glucose content and placental GLUT3 glucose transporter expression, but does not obviously affect fetal plasma glucose concentration. Moreover, the present results are not compatible with a role of maternal corticosterone in the development of growth-restricted rat fetuses.


[Abdominal lymphoma with abscess and thoracic perforation: en-bloc resection].
The authors compare a rare case of acute onset Gastric Lymphoma with that present in the literature. The patient, a white 67 year-old man, was admitted to hospital in severe general condition. On CT scan an opacity of the pleura and a subtotal collapse of the left lung associated with a mass infiltrating the diaphragm, the gastric fundus and pancreas were seen. After 48 hours from the admission an operation was performed. The abdominal mass infiltrating the stomach, the spleen and the left thorax was resected en-bloc by laparotomy. A Boulau drainage was inserted. The histology showed an high malignant, diffused, big cell lymphoma. The patient was discharged 23 days later after a regular postoperative course. Surgery can be necessary, sometime curative in gastric lymphomas while long term result and quality of life could be improved by chemotherapy.


Gait patterns after anterior cruciate ligament reconstruction are related to graft type.
Although there is a tendency toward gait normalization after anterior cruciate ligament reconstruction, altered moments about the knee flexion-extension axis have been reported. It is possible that these gait alterations relate to donor site morbidity associated with the graft harvest. There is a relationship between graft type and external knee moments during walking. Controlled laboratory study. Three groups were compared: 17 patellar tendon anterior cruciate ligament reconstruction patients (mean, 11 months after surgery), 17 hamstring tendon anterior cruciate ligament reconstruction patients (mean, 9.3 months after surgery), and 17 matched controls. A 3-dimensional motion analysis and force plate system was used to determine sagittal plane kinematics and kinetics of the lower limb during comfortable-speed walking. There were significant differences in the moments about the knee that related to graft type. The external knee flexion moment at midstance was significantly smaller than that in the control knees in 65% of patients in the patellar tendon group and 29% of patients in the hamstring tendon group. In contrast, the external knee extension moment at terminal stance was significantly smaller than that in the control knees in 53% of subjects in the hamstring tendon group and 23% of subjects in the patellar tendon group. There are graft-specific differences in knee biomechanics after anterior cruciate ligament reconstruction that appear to relate to the donor site. Considerable debate continues as to whether the patellar tendon or the hamstring tendon graft is preferable for anterior cruciate ligament reconstruction. It is therefore clinically relevant to understand the biomechanical differences in knee function associated with both graft types.


Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface.
Bacterial surface-layers (S-layers) are crystalline arrays of repeating proteinaceous subunits that coat the exterior of many cell envelopes. S-layers have demonstrated diverse functions in growth and survival, maintenance of cell integrity, and mediation of host interactions. Additionally, S-layers can act as scaffolds for the outward display of auxiliary proteins and glycoproteins. These non-covalently bound S-layer associated proteins (SLAPs) have characterized roles in cell division, adherence to intestinal cells, and modulation of the host immune response. Recently, IgdA (LBA0695), a Lactobacillus acidophilus SLAP that possesses a Group 3 immunoglobulin (Ig)-like domain and GW (Gly-Tryp) dipeptide surface anchor, was recognized for its high conservation among S-layer-forming lactobacilli, constitutive expression, and surface localization. These findings prompted its selection for examination within the present study. Although IgdA and corresponding orthologs were shown to be unique to host-adapted lactobacilli, the Ig domain itself was specific to vertebrate-adapted species suggesting a role in vertebrate adaptation. Using a counterselective gene replacement system, igdA was deleted from the L. acidophilus NCFM chromosome. The resultant mutant, NCK2532, exhibited a visibly disrupted cell surface which likely contributed to its higher salt sensitivity, severely reduced adhesive capacity, and altered immunogenicity profile. Transcriptomic analyses revealed the induction of several stress response genes and secondary surface proteins. Due to the broad impact of IgdA on the cellular physiology and probiotic attributes of L. acidophilus, identification of similar proteins in alternative bacterial species may help pinpoint next-generation host-adapted probiotic candidates.


Analysis of intrastrain recombination in herpes simplex virus type 1 strain 17 and herpes simplex virus type 2 strain HG52 using restriction endonuclease sites as unselected markers and temperature-sensitive lesions as selected markers.
The viral and host factors involved in herpes simplex virus (HSV) recombination are little understood. To identify features of the process, recombination in HSV-1 and HSV-2 has been studied by analysing the segregation of unselected markers in the form of restriction endonuclease (RE) sites. By confining parental interactions to only one strain of virus of each serotype, restrictions imposed by non-homology are overcome and differential growth phenotypes can be discounted. The analysis of unselected and selected recombinants using RE sites in conjunction with temperature-sensitive mutations is consistent with (i) HSV being highly recombinogenic, (ii) parental and progeny molecules taking part in the process, (iii) the four genomic isomers participating in recombination, (iv) genome alignment being part of the recombination process and (v) cellular factors in conjunction with genome homology influencing the efficiency of recombination.


Improving fat transfer viability: the role of aging, body mass index, and harvest site.
Fat grafting remains unpredictable in the clinical setting, and variables that influence adipocyte survival, such as age, body mass index, and specific donor sites, are still not well understood. Twenty-four female subjects were enrolled in this research after institutional review board approval and signed consent to participate was obtained. Subjects were separated into groups according to (1) age (younger, ≤45 years; and older, ≥46 years) and (2) body mass index (normal weight, body mass index<25; and overweight, body mass index≥25). All fat samples were obtained through dry liposuction of three donor sites: lower abdomen, inner thigh, and flank. They were processed identically for dissociation of adipose tissue and isolation of adipocytes. Adipocyte viability was measured using the Nexcelom Cellometer Auto T4. In younger patients, adipocyte viability was greater in the lower abdomen than in the flank; in older patients, this difference was not seen. When lower abdominal fat from younger was compared with that from older patients, the viability was higher in younger patients. However, adipocytes from the flank depot had higher viability in the older group compared with the younger group. Inner thigh fat viability was not significantly different across the two age groups. The authors also found no significant differences in fat viability for any given donor site between the normal weight and overweight body mass index groups. The optimal choice site for fat harvest should take patient age into consideration. In younger patients, both lower abdomen and inner thigh appear to be good options. Therapeutic, II.


[The role of hyperpolarization and depolarization of the membrane of the human spermatozoon].
The electric potential across the membrane (psi) is evaluated through the accumulation of the lipophilic, radiolabelled, triphenylmethylphosphonium (3H-TPMP+), cation on membrane human sperm. The washed sperm cells, were incubated in presence of 3H-TPMP+ in low-K+ medium or high-K+ and allowed to take up the cation to steady-state (ie; 20 min at 37 degrees C). By using this differential, the value obtained was inserted in the Nernst equation, this value yielded a psi of -69 +/- 2 mV. The presence of divalent cations Zn++ and Mg++ in the incubation medium both induced a hyperpolarization of 10% and 8.6%, respectively. The addition of specific reagents such as p-chloromercuribenzene-sulfonate and ethylene-diamine-tetraacetic acid sodium salt both decreased the psi 35% and 58% respectively. The agents to act upon the components of the sperm cell membrane such as the dithiothreitol and progesterone induced hyperpolarization and depolarization of the membrane 16% and 40%, respectively. The presence of propranolol and L-alpha-lysophosphatidylcholine, which affect the ionic gradients present across the plasma membrane, both induced a depolarization from 43% and 92% respectively. Finally, the addition of tetraphenylboron (TPB-) on the incubation medium, enhanced the value of the psi 75%. These studies are transcendent because with the utilization of agents depolarizing, or hyperpolarizing we obtain changes in the psi from -80 +/- 0.6 mV, until -6 +/- 0.6 mV changes of 74 +/- 1.5 mV across of the sperm cell membrane.


In situ TEM of biological assemblies in liquid.
Researchers regularly use Transmission Electron Microscopes (TEMs) to examine biological entities and to assess new materials. Here, we describe an additional application for these instruments- viewing viral assemblies in a liquid environment. This exciting and novel method of visualizing biological structures utilizes a recently developed microfluidic-based specimen holder. Our video article demonstrates how to assemble and use a microfluidic holder to image liquid specimens within a TEM. In particular, we use simian rotavirus double-layered particles (DLPs) as our model system. We also describe steps to coat the surface of the liquid chamber with affinity biofilms that tether DLPs to the viewing window. This permits us to image assemblies in a manner that is suitable for 3D structure determination. Thus, we present a first glimpse of subviral particles in a native liquid environment.


Comparison of perceived exertion ratings during graded peak cycle exercise between cigarette smokers and nonsmokers.
This study compared differentiated Ratings of Perceived Exertion (RPE) for the chest (RPE-Chest) and legs (RPE-Legs) and undifferentiated ratings (RPE-Overall) between cigarette smokers (n = 10) and nonsmokers (n = 10) during graded, peak cycle exercise. Linear regression analyses expressed RPE-Chest, RPE-Leg, and RPE-Overall as a function of VO2 for smokers and nonsmokers. RPE equivalent to 50, 70, and 90% VO2 peak were compared between. Chest, Leg, and Overall RPE were similar at each % VO2 peak. In the combined smokers and nonsmokers, RPE-Chest was lower (p < .05) than the RPE-Overall at each % VO2 peak. RPE-Leg did not differ from RPE-Overall at 50% VO2 peak, but was higher (p < .05) at 70% and 90% VO2 peak. These results validate Assumption 4 for application of the Borg 15-category scale in that this assumption is also valid for differentiated RPE. Smoking did not influence the intensity of differentiated or undifferentiated RPE during graded cycle exercise.


The Accuracy of Clinical Diagnosis in 2135 Lesions on the Face. A Retrospective Analysis of Histopathological Records.
Biopsy of facial skin lesions is an important supplement to dermatological diagnostics, especially in doubtful cases or suspected of being malignant. The aim of the retrospective study of 2135 histopathological records of lesions on the face was to: establish the most common indications for a skin biopsy in patients with facial lesions, establish the frequency of histopathological diagnoses, evaluate how often clinically suspected inflammatory lesions are identified as tumors in histopathology, evaluate the accuracy of clinical diagnoses of the most common skin tumors and dermatoses. It was a retrospective study. Histopathological records from the lesions on the face from years 2010-2017 were analyzed. The mean age of patients was 69.3 [7-98]. Fifty-eight percent of the patients were women. Among 2135 clinical diagnoses skin tumors were suspected in 1905 cases. Among 2169 obtained histopathological results (34 biopsies showed 2 diseases), we identified skin tumors in 1940 cases, with 1388 confirmed as malignant. The clinical diagnosis of a specific benign or malignant skin tumor was accurate in 1013/1634 subjects, in comparison to inflammatory lesions, which were correct in 67/148 cases, (P = 0.0001). Among all preliminary inflammatory diagnoses, 33/204 lesions were identified as skin tumors in histopathology. In conclusion in most cases of skin tumors the clinical diagnosis is confirmed by histopathological examination. In case of facial inflammatory lesions, the accuracy of clinical diagnosis is lower, with a significant number of facial lesions appearing inflammatory in clinical evaluation but being diagnosed as skin cancers in pathology.


Alcohol abuse in adolescents.
Alcohol abuse by adolescents is widespread in the United Kingdom, has complex cultural, economic and biological origins and is associated with adverse health consequences. Affected young people present medically with intoxication or its immediate consequences such as injury or self-harm. A significant number also suffer adverse psychosocial effects and longer-term adverse outcomes. Even some younger patients are now presenting with evidence of physiological dependence. Interpretation of the literature and clinical experience suggests that practitioners have greater capacity to respond effectively than they may believe. Modest concerted effort could make the medical profession much more relevant to the needs of these, often troubled, young people.


Erythrocyte permeability to lipophilic solutes changes with temperature.
Studies of permeability coefficients of biological barriers to members of homologous series can provide information of value in assessing barrier characteristics. To this end, we have determined the linear diffusion coefficients of tracer water (THO), [14C]antipyrine, [14C]acetamide, and n-[14C]alcohols over the range of 10-37 degrees C for dog erythrocytes (D), hemoglobin (D2), and plasma (D1). Permeability coefficients (Po) calculated with the series-parallel pathway model are higher than Po for water at 37 degrees C for all the alcohols except hexanol. Po for acetamide and antipyrine are considerably lower than Po for water at 37 degrees C. The apparent activation energies (Ea) for Po of acetamide and the C1, C2, and C3 alcohols are 10.5 kcal.mol-1, similar to the values obtained with epithelial or lipid bilayers. The Ea's for the larger alcohols are 2-4 kcal.mol-1. The Ea for Po of water is 5.3 kcal.mol-1, similar to the Ea for self-diffusion of water; Ea for antipyrine is 25 kcal.mol-1. We interpret these results to indicate heterogeneity of membrane-solute interactions or of membrane pathways in the erythrocyte for lipid-soluble molecules that is related to both lipid solubility and solute size, as we have suggested in an earlier study and confirmed experimentally in the present one.


Surface hopping with a manifold of electronic states. II. Application to the many-body Anderson-Holstein model.
We investigate a simple surface hopping (SH) approach for modeling a single impurity level coupled to a single phonon and an electronic (metal) bath (i.e., the Anderson-Holstein model). The phonon degree of freedom is treated classically with motion along--and hops between--diabatic potential energy surfaces. The hopping rate is determined by the dynamics of the electronic bath (which are treated implicitly). For the case of one electronic bath, in the limit of small coupling to the bath, SH recovers phonon relaxation to thermal equilibrium and yields the correct impurity electron population (as compared with numerical renormalization group). For the case of out of equilibrium dynamics, SH current-voltage (I-V) curve is compared with the quantum master equation (QME) over a range of parameters, spanning the quantum region to the classical region. In the limit of large temperature, SH and QME agree. Furthermore, we can show that, in the limit of low temperature, the QME agrees with real-time path integral calculations. As such, the simple procedure described here should be useful in many other contexts.


Effects of carnosine on volatile generation from Maillard reaction of ribose and cysteine.
Carnosine occurs naturally in meat and meat products in significant quantity, and it possesses strong antioxidant activity that inhibits lipid oxidation and enhances shelf life. In this study, the effects of carnosine on thermal flavor generation were investigated using the model system of cysteine and ribose, which was heated to the roasting temperature of 180 degrees C for 2 h at pH 5 and pH 8.5. The results indicated that carnosine affected volatile formation in a complex manner. Volatiles identified from the liquid phase of the reaction systems of ribose and cysteine showed that the sulfur-containing compounds such as thiophenes, thiazoles, and polysulfides were the most abundant compounds. The addition of carnosine into the reaction mixtures in general caused a reduction in contents of thiophenes and some important meaty flavor compounds such as 2-methyl-3-furanthiol, 2-furfurylthiol, and their associated dimers. On the other hand, it facilitated the generation of several important nitrogen-containing volatiles such as pyrazine, methylpyrazine, 2,6-dimethylpyrazine, and other alkyl pyrazines and thiazoles, which are known to elicit roasty and nutty flavor notes. The results suggested that carnosine acts as a nitrogenous source to facilitate the formation of nitrogen-containing compounds, possibly by degradation to form ammonia.


The future financing of Medicare.
Regardless of the timing and the type of federal action to reform the nation's health care system, the Medicare program's financial problems must be addressed soon. Serious concerns exist about both the Hospital Insurance and the Supplementary Medical Insurance portions of the program.


[Various quantitative characteristics of the pathoarchitectonics of the cerebral cortex in Pick disease and Alzheimer disease].
On the basis of the study of the pathological architectonics of 6 brain hemispheres in Pick's disease (PD) and 4 hemispheres in Alzheimer's disease (AD), the author presents data on changes in the size of the surface of the hemispheres on the whole and of individual areas and compares them with the mean figures. It is concluded that the degenerative, atrophic process in both groups of presenile dementia involves the entire brain cortex: the indices of the area of the entire hemisphere in all the cases studied were below the mean values. The surface of the cortex was shrinked in both phylogenetically new areas (inferior occipital, frontal and temporal) and phylogenetically older regions (pre- and postcentral, occipital), as well as in the limbic and insular regions. In all cases the general regularity was maintained: a decrease in the size of the entire hemispherical surface as well as individual regions was more expressed in AD than in PD. The exception to this rule was 1 case of PD in which all parameters were very low which may be associated with the duration of the disease (12 years). There was a certain asymmetry between the size of the hemisphere on the whole and of its individual regions. No correlation was found between the degree of macroscopic atrophy of the convolutions and the severity of pathological achitectonic changes in cortical formations.


The last glacial-Holocene transition in southern Chile.
Warming at the last glacial termination in the North Atlantic region was interrupted by a period of renewed glacial activity during the Younger Dryas chronozone (YDC). The underlying mechanism of this cooling remains elusive, but hypotheses turn on whether it was a global or a North Atlantic phenomenon. Chronological, sedimentological, and palaeoecological records from sediments of small lakes in oceanic southern Chile demonstrate that there was no YDC cooling in southern Chile. It is therefore likely that there was little or no cooling in southern Pacific surface waters and hence that YDC cooling in the North Atlantic was a regional, rather than global, phenomenon.


Asthma in children born after infertility treatment: findings from the UK Millennium Cohort Study.
Is asthma more common in children born after subfertility and assisted reproduction technologies (ART)? Yes. Asthma, wheezing in the last year and anti-asthmatic medication were all more common in children born after a prolonged time to conception (TTC). This was driven specifically by an increase in children born after ART. Few studies have investigated any association between ART and asthma in subsequent children, and findings to date have been mixed. A large registry-based study found an increase in asthma medication in ART children but suggests underlying infertility is the putative risk factor. Little is known about asthma in children after unplanned or mistimed conceptions. The Millennium Cohort Study is a UK-wide, prospective study of 18 818 children recruited at 9 months of age. Follow-up is ongoing. This study analyses data from follow-up surveys at 5 and 7 years of age (response rates of 79 and 70%, respectively). Singleton children whose natural mothers provided follow-up data were included. Mothers reported whether their pregnancy was planned; planners provided TTC and details of any ART. The population was divided into 'unplanned' (unplanned and unhappy), 'mistimed' (unplanned but happy), 'planned' (planned, TTC < 12 months), 'untreated subfertile' (planned, TTC >12 months), 'ovulation induced' (received clomiphene citrate) and 'ART' (IVF or ICSI). The primary analysis used the planned children as the comparison group; secondary analysis compared the treatment groups to the children born to untreated subfertile parents. Outcomes were parent report of asthma and wheezing at 5 and 7 years, derived from validated questions in the International Study of Asthma and Allergies in Childhood, plus use of anti-asthmatic medications. A total of 13 041 (72%) children with full data on asthma and confounders were included at 5 years of age, and 11 585 (64%) at 7 years. Compared with planned children, those born to subfertile parents were significantly more likely to experience asthma, wheezing and to be taking anti-asthmatics at 5 years of age [adjusted odds ratio (OR): 1.39 (95% confidence interval (CI): 1.07, 1.80), OR: 1.27 (1.00, 1.63) and OR: 1.90 (1.32,2.74), respectively]. This association was mainly related to an increase among children born after ART (adjusted OR: 2.65 (1.48, 4.76), OR: 1.97, (1.10, 3.53) and OR: 4.67 (2.20, 9.94) for asthma, wheezing and taking anti-asthmatics, respectively). The association was also present, though reduced, at the age of 7 years. The number of singletons born after ART was relatively small (n = 104), and as such the findings should be interpreted with caution. However, data on a wide range of possible confounding and mediating factors were available and analysed. The data were weighted for non-response to minimize selection bias. The findings add to the growing body of evidence suggesting an association between subfertility, ART and asthma in children. Further work is needed to establish causality and elucidate the underlying mechanism. These findings are generalizable to singletons only, and further work on multiples is needed.


[Risk indicators of preventable morbidity related to drug utilization].
To select clinical situations that can be used as risk indicators of preventable morbidity caused by drugs at the community pharmacies, and to study their acceptability, in terms of pertinence and relevance. We used the Delphi technique, in 2 rounds, by a panel of 14 medical doctors and pharmacists experts, to study the relevance of 68 types of clinical situations as risk indicators of preventable morbidity related to drug utilization used by health professionals in community pharmacies, with scientific evidence of foreseeable adverse result, frequent situations in ambulatory care and with controllable cause and result. 43 of the 68 indicators were considered usable and pertinent. The indicators referred three areas: drug type (medications of narrow therapeutic margin, with individualized dose and adverse reactions frequent and severe), health problem (chronic problems, especially asthma, cardiac, thyroid and prostate illness, and pain), and patient (old or with several medications. Pharmacists systematically overvalued some indicators in relation to the doctors, but differences were not significant. Forty-three indicators were selected as valuable to identify situations of preventable morbidity related to drug utilization.


Ultrasound avoidance by flying antlions (Myrmeleontidae).
The acoustic arms race between insectivorous bats and their invertebrate prey has led to the convergent evolution of ultrasound hearing in seven orders of nocturnal insects. Upon hearing the echolocation calls of an approaching bat, such insects take defensive action. Here, we document a previously unknown sense of ultrasound hearing and phonotactic flight behaviour in the neuropteran family Myrmeleontidae (antlions). The antlion Myrmeleon hyalinus was presented with sound pulses at ultrasonic frequencies used by echolocating bats and its response thresholds in tethered flight determined. Behaviours included abdominal twitches, wing flicks, brief pauses in flight and flight cessation. Such behaviours create erratic evasive flight manoeuvres in other eared insects, particularly mantids and lacewings. Antlions responded best to ultrasound between 60 and 80 kHz (75 dB peSPL at 80 kHz), showing response thresholds similar to those of the related lacewings (Neuroptera, Chrysopidae). Yet, at lower ultrasonic frequencies (20-50 kHz), antlions were far less sensitive than lacewings. Based on calculated response distances, we conclude that antlions respond only after having been detected by bats rather than using early evasive flights. We argue that the high response threshold for low-frequency ultrasound is adaptive for an insect that is mainly active close to and within vegetation, because a behavioural response to the lower ultrasonic frequencies used by high-flying bats would result in evasive action in the absence of actual predation risk.


Self-perceived knowledge and skills of job coaches in Japan.
The purpose of the study was to assess the self-perceived knowledge and skills of Japanese job coaches and to examine whether their knowledge and skills differed across employment settings. The 479 job coaches at Work Support Centers or Work Support Agencies comprised the study population. A Japanese version of the 80-item Self-Assessment for Students or Counselors (SASC-J) was mailed to all the Work Support Centers and Agencies. There was no significant difference on any of the SASC-J 8 subsystems mean scores between Work Support Agencies and Work Support Centers. The highest mean score of these 2 employment settings was the "Placement Personal" (2.30 and 2.31), and the lowest was the "Education" (1.40 and 1.46). The overall mean score of the SASC-J was 1.82 (SD=0.63). A significant relationship was found between the years of experience and the SASC-J (r=0.30, p < 0.01). Since the average below 3.0 on the SASC would mean that "you need to read a textbook on placement and/or a course in Placement", the result of the current study suggested that Japanese job coaches, regardless of the employment settings, need to learn more about the systematic placement technique. Further studies are encouraged to assess the training outcome of the job coach.


Local electronic descriptors for solute-defect interactions in bcc refractory metals.
The interactions between solute atoms and crystalline defects such as vacancies, dislocations, and grain boundaries are essential in determining alloy properties. Here we present a general linear correlation between two descriptors of local electronic structures and the solute-defect interaction energies in binary alloys of body-centered-cubic (bcc) refractory metals (such as W and Ta) with transition-metal substitutional solutes. One electronic descriptor is the bimodality of the d-orbital local density of states for a matrix atom at the substitutional site, and the other is related to the hybridization strength between the valance sp- and d-bands for the same matrix atom. For a particular pair of solute-matrix elements, this linear correlation is valid independent of types of defects and the locations of substitutional sites. These results provide the possibility to apply local electronic descriptors for quantitative and efficient predictions on the solute-defect interactions and defect properties in alloys.


Whole genome amplification of single cells from clinical peripheral blood smears.
Molecular analysis of clinical samples has been hampered by the lack of fresh or frozen specimens and the presence of contaminating background cells within samples obscuring the molecular analysis of the pathological cells of interest. Routine cytology specimens are a ubiquitous and abundant, yet largely untapped, source of clinical samples for molecular analysis. Morphologically defined single cells from peripheral blood smears can be microdissected from contaminating background cells and their whole genome amplified by primer extension preamplification, followed by polymerase chain reaction analysis of the specific DNA of interest. Thus, molecular information can be traced back to the cell of origin in these clinical specimens. This should allow studies on clonality, loss of heterozygosity, mutation, or amplification of multiple loci from one single cell in haematological smears and possibly other clinical cytology specimens.


Laparoscopic common bile duct exploration.
In recent years, laparoscopic common bile duct exploration has become the procedure of choice in the management of choledocholithiasis in several laparoscopic centers. The increasing interest for this laparoscopic approach is due to the development of instrumentation and technique, allowing the procedure to be performed safely, and it is also the result of the revised role of endoscopic retrograde cholangiopancreatography, which has been questioned because of its cost, risk of complications and effectiveness. Many surgeons, however, are still not familiar with this technique. In this article we discuss the technique and results of laparoscopic common bile duct exploration. Both the laparoscopic transcystic approach and choledochotomy are discussed, together with the results given in the literature. When one considers the costs, morbidity, mortality and the time required before the patient can return to work, it would appear that laparoscopic cholecystectomy with common bile duct exploration is more favorable than open surgery or laparoscopic cholecystectomy with preoperative or postoperative endoscopic sphincterotomy. However, the technique requires advanced laparoscopic skills, including suturing, knot tying, the use of a choledochoscope, guidewire, dilators and balloon stone extractor. Although laparoscopic common bile duct exploration appears to be the most cost-effective method to treat common bile duct stones, it should be emphasized that this procedure is very challenging, and it should be performed by well-trained laparoscopic surgeons with experience in biliary surgery.


[Preliminary functional analysis of intron in chalcone synthase gene from Scutellaria baicalensis].
To study the function of the chalcone synthese gene introns in Scutellaria baicalensis, and clarify preliminarily their role in abiotic stress. The CHS introns with specific primers were cloned and bioinformatic method was applied to predict the cis-elements in the intron of CHS. The introns were subcloned into binary vector, pCAMBIA-1301 before being transferred to tobacco. Then the activity of GUS of the transgenic tobacco seeds was analyzed. Seven cis-elements were found in the introns. Under the dark and high temperature GUS expression rose at the first (3 h), but then declined (9 h). ABA and MeJA regulated insignificantly the GUS activity in normal temperature; treatment of 10% PEG induced GUS expression. CHS introns could be play a role in the regulation of S. baicalensis phenylpropanoid biosynthetic pathway.


Neuromagnetic localization of epileptiform spike activity in the human brain.
Local paroxysmal discharges of epileptic tissue within the human brain, which may be electrically recorded as voltage spikes in the electroencephalogram, also generate extracranial magnetic fields. These fields were assessed by means of recently developed neuromagnetometric techniques. Surface measurements of magnetic spike field strength in the region of the focus appear sufficient to establish the location, depth, orientation, and polarity of currents underlying the paroxysmal discharge.


Sleep duration associated with cardiovascular conditions among adult Nevadans.
Associations between sleep duration and cardiovascular conditions have been inconsistent. Both short and long sleep duration are associated with increased risk for diabetes, hypertension, obesity, high cholesterol, depression, and overall metabolic dysfunction; all of which are cardiovascular disease risk factors. This study aimed to determine if deficient or excessive sleep duration is associated with cardiovascular conditions. Data were obtained from the 2013 Behavioral Risk Factor Surveillance System for the state of Nevada. A total of 5101 participants completed the survey and answered questions as to whether they had ever had a cardiovascular condition ie, myocardial infarction, coronary heart disease or angina, and stroke. Multiple logistic regression was implemented during analyses and yielded four models including demographics, co-morbidity, behavior, and final significant variables. Six significant predictor variables were identified in the final model. Sleep duration was statistically significant (p < 0.0001). Individuals having deficient sleep, eg, 1-4 h a night, were 2.4 times more likely to have a cardiovascular condition versus those sleeping 7-9 h per night (OR = 2.412, CI = 1.139, 5.107). As for individuals who sleep excessively, eg, 10-18 h a night, they were nearly 7.2 times more likely to have a cardiovascular condition, compared to individuals who receive a normal night's sleep (OR = 7.170, CI = 3.284, 15.654). Both deficient and excessive sleep duration were significantly associated with a cardiovascular condition, even after adjustment for covariates. The findings from this study can be used as additional evidence to encourage further research on improving sleep by developing future treatment therapies, and recommendations, that may help lower the risk of cardiovascular disease conditions.


PDGF-BB-induced DNA synthesis is delayed by angiotensin II in vascular smooth muscle cells.
The interaction of ANG II with platelet-derived growth factor (PDGF)-BB-induced DNA synthesis was studied in cultured rat aortic smooth muscle cells. PDGF-BB-induced DNA synthesis was delayed (approximately 6-8 h) by ANG II as shown by a time-course experiment. Losartan, an AT1-receptor antagonist, blocked the transient inhibitory effect of ANG II, whereas the AT2-receptor antagonist PD-123319 had no effect. Autocrine- or paracrine-acting transforming growth factor-beta 1 (TGF-beta 1), believed to be a mediator of ANG II-induced inhibitory effects, was not responsible for the delay of PDGF-BB-induced DNA synthesis, because a potent TGF-beta 1 neutralizing antibody could not reverse this effect of ANG II, nor was the delay of the PDGF-BB effect caused by inhibition of PDGF-beta-receptor phosphorylation as shown by Western blot analysis of immunoprecipitated PDGF-beta receptor. In conclusion, our results show that ANG II can exert a transient inhibitory effect on PDGF-BB-induced proliferation via the AT1 receptor.


A Review of the Analgesic Benefi ts and Potential Complications Related to Epidural Corticosteroid Injections.
The Institute of Medicine has reported that greater than 115 million adults in the United States are living with some form of chronic pain. Back pain is the most prevalent and is associated with high individual morbidity and increased healthcare costs. One approach for the management of chronic back pain involves the injection of corticosteroids in the epidural space.This interventional approach requires advanced training with techniques that vary according to the level of the vertebral column where the injection is to be performed. The primary rationale for epidural steroid injection is to reduce the inflammation surrounding the spinal nerve root as it exits the neuroforamen.Injections are performed at levels that correspond most appropriately with the patient's clinical presentation,physical findings, and radiographic findings. Epidural steroid injections are considered safe and effective, and are supported by evidence for the treatment of radicular pain. Complications from epidural steroid injections are rare but can be catastrophic, including permanent disability and death. The focus of this article is to understand how technique and selection of specific corticosteroids used for epidural injection can manage chronic back and radicular pain effectively while minimizing risk that leads to unnecessary harm.


Round window electrocochleography just before cochlear implantation: relationship to word recognition outcomes in adults.
Electrocochleography (ECoG) to acoustic stimuli can differentiate relative degrees of cochlear responsiveness across the population of cochlear implant recipients. The magnitude of the ongoing portion of the ECoG, which includes both hair cell and neural contributions, will correlate with speech outcomes as measured by results on CNC word score tests. Postoperative speech outcomes with cochlear implants vary from almost no benefit to near normal comprehension. A factor expected to have a high predictive value is the degree of neural survival. However, speech performance with the implant does not correlate with the number and distribution of surviving ganglion cells when measured postmortem. We will investigate whether ECoG can provide an estimate of cochlear function that helps predict postoperative speech outcomes. An electrode was placed at the round window of the ear about to be implanted during implant surgery. Tone bursts were delivered through an insert earphone. Subjects included children (n = 52, 1-18 yr) and postlingually hearing impaired adults (n = 32). Word scores at 6 months were available from 21 adult subjects. Significant responses to sound were recorded from almost all subjects (80/84 or 95%). The ECoG magnitudes spanned more than 50 dB in both children and adults. The distributions of ECoG magnitudes and frequencies were similar between children and adults. The correlation between the ECoG magnitude and word score accounted for 47% of the variance. ECoGs with high signal-to-noise ratios can be recorded from almost all implant candidates, including both adult and pediatric populations. In postlingual adults, the ECoG magnitude is more predictive of implant outcomes than other nonsurgical variables such as duration of deafness or degree of residual hearing.


Wide disparity in genetic admixture among Mexican Americans from San Antonio, TX.
We studied 706 participants of the San Antonio Family Diabetes Study (SAFDS) and 586 male samples from the San Antonio Center for Biomarkers of Risk of Prostate Cancer (SABOR) and used 64 ancestry informative markers to compare admixture proportions between both groups. Existence of population substructure was demonstrated by the excess association of unlinked markers. In the SAFDS sample, ancestral proportions were estimated at 50.2 ± 0.6% European, 46.4 ± 0.6% Native American, and 3.1 ± 0.2% West African. For the SABOR sample, the proportions were 58.9 ± 0.7%, 38.2 ± 0.7%, and 2.9 ± 0.2%, respectively. Additionally, in the SAFDS subjects a highly significant negative correlation was found between individual Native American ancestry and skin reflectance (R(2) = 0.07, P= 0.00006). The correlation was stronger in males than in females but clearly showed that ancestry only accounts for a small percentage of the variation in skin color and, conversely, that skin reflectance is not a robust surrogate for genetic admixture. Furthermore, a substantial difference in substructure is present in the two cohorts of Mexican American subjects from the San Antonio area in Texas, which emphasizes that genetic admixture estimates should be accounted for in association studies, even for geographically related subjects.


The low-dose ACTH stimulation test: is 30 minutes long enough?
Controversy persists regarding the use of the low-dose adrenocorticotropic hormone (ACTH) stimulation test (LDST) for the diagnosis of adrenal insufficiency (AI) and optimal test result interpretation. However, many centers are now using the LDST to assess cortisol secretion adequacy, and some only use a 30-minute cortisol level to determine adrenal sufficiency or AI. This study examined both 30- and 60-minute cortisol levels to assess whether the interpretation of the test was affected when both cortisol levels were taken into consideration. Data were obtained by retrospective chart review from a single pediatric endocrinology unit over a 7-year period. We identified 82 patients who completed the LDST. Their mean age was 11.7 years, and 37% were female. Cortisol levels were evaluated at baseline and 30 and 60 minutes after cosyntropin administration. A cutoff value ≥18 μg/dL was used to define adrenal sufficiency. We found that 54% of patients reached peak cortisol levels at 60 minutes, and 11 patients who did not pass the test at 30 minutes did so at 60 minutes. The only predictive characteristic was weight status; overweight and obese individuals tended to peak at 30 minutes, and normal and underweight individuals tended to peak at 60 minutes. Although further studies are necessary to confirm our findings, it appears that measuring cortisol both 30 and 60 minutes following synthetic ACTH administration may be necessary to avoid overdiagnosing AI.


Effects of psychotropic drugs on Nuclear Factor kappa B.
Mental disorders affect millions of people worldwide and are associated with a huge suffering and unbearable burden for patients and their care givers. The pathophysiology of mental disorders is not fully understood. In recent years, accumulating data suggest that inflammation may play a role in the pathogenesis of these illnesses and that psychotropic drugs exert some anti-inflammatory effects. Nuclear Factor κB (NF-κB) is a cellular pathway that has a prominent influence on immune and inflammatory responses in humans. Numerous studies examined the effects of psychotropic drugs on different inflammatory mediators (particularly cytokines) both in vitro and in vivo. However, relatively few studies investigated the effects of those drugs on NF-κB. This is quite surprising considering the pivotal role of NF-κB in promoting inflammation. The aim of this article is to review the data over the effects of psychotropic drugs on the NF-κB pathway. Overall, the summarized studies suggest that some psychotropic drugs (such as lithium and imipramine) exert potent inhibitory effects on NF-κB, while the results on other drugs are not conclusive and occasionally contradicting. The discrepancy in the results of different studies seems to derive from the various experimental conditions under which the drugs were tested.


Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.
The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system. The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV). Three samples with sperm concentration of about 100 × 106 /mL were diluted to evaluate the linear range. The detection values of latex beads were within the range of target values. The CVs of sperm concentration and PR were significantly and negatively correlated with sperm concentration (r = -0.561, p = 0.001) and PR value (r = -0.621, p < 0.001), respectively. The R2 values of the linear range of sperm concentration were ≥0.99. There was no significant difference in sperm motility and PR within 1-10 min at 36.5°C ± 0.5°C. The coincidence rates of sperm morphology and sperm head morphology for 36 semen samples analyzed by the GSA-810 system and manual method were 99.40% and 99.67%, respectively. The CVs of the percentage of sperm with abnormal morphology and percentage of sperm with abnormal head morphology were less than 5%. The GSA-810 system can accurately analyze normal semen samples, but the repeatability of the results is poor for oligozoospermia and asthenozoospermia samples. The future CASA system for analyzing sperm morphology should focus on recognizing the middle and tail segments of a spermatozoon.


Effect of total leucocyte count on whole blood filterability in patients with peripheral vascular disease.
An abnormal filterability of whole blood through micropore membranes in vitro has been reported in peripheral vascular disease and has been thought to indicate abnormal red cell deformability. Blood from 68 patients with symptomatic peripheral vascular disease of varying severity and from 32 age-matched controls without a history of peripheral vascular disease was studied by the technique of whole blood filtration. In agreement with earlier findings, whole blood filterability was significantly reduced in patients with symptoms of vascular disease, but also their total leucocyte count was significantly higher than that of the controls. Variation in leucocyte count was found to affect significantly whole blood filterability, and the abnormal filterability in peripheral vascular disease could be entirely ascribed to this factor and not to an alteration in red cell deformability. The raised leucocyte count was not due to smoking but its cause could not be explained.


Efficient Remyelination Requires DNA Methylation.
Oligodendrocyte progenitor cells (OPCs) are the principal source of new myelin in the central nervous system. A better understanding of how they mature into myelin-forming cells is of high relevance for remyelination. It has recently been demonstrated that during developmental myelination, the DNA methyltransferase 1 (DNMT1), but not DNMT3A, is critical for regulating proliferation and differentiation of OPCs into myelinating oligodendrocytes (OLs). However, it remains to be determined whether DNA methylation is also critical for the differentiation of adult OPCs during remyelination. After lysolecithin-induced demyelination in the ventrolateral spinal cord white matter of adult mice of either sex, we detected increased levels of DNA methylation and higher expression levels of the DNA methyltransferase DNMT3A and lower levels of DNMT1 in differentiating adult OLs. To functionally assess the role of DNMT1 and DNMT3 in adult OPCs, we used mice with inducible and lineage-specific ablation of Dnmt3a and/or Dnmt1 (i.e., Plp-creER(t);Dnmt3a-flox, Plp-creER(t);Dnmt1-flox, Plp-creER(t);Dnmt1-flox;Dnmt3a-flox). Upon lysolecithin injection in the spinal cord of these transgenic mice, we detected defective OPC differentiation and inefficient remyelination in the Dnmt3a null and Dnmt1/Dnmt3a null mice, but not in the Dnmt1 null mice. Taken together with previous results in the developing spinal cord, these data suggest an age-dependent role of distinct DNA methyltransferases in the oligodendrocyte lineage, with a dominant role for DNMT1 in neonatal OPCs and for DNMT3A in adult OPCs.


A point mutation of human nucleoside diphosphate kinase A found in aggressive neuroblastoma affects protein folding.
The point mutation serine 120 to glycine in the human nucleoside diphosphate kinase A has been identified in several aggressive neuroblastomas (Chang, C. L., Zhu, X. X., Thoraval, D. H., Ungar, D., Rawwas, J., Hora, N., Strahler, J. R., Hanash, S. M. & Radany, E. (1994) Nature 370, 335-336). We expressed in bacteria and purified wild-type and S120G mutant nucleoside diphosphate kinase A. The mutant enzyme had enzymatic and structural properties similar to the wild-type enzyme, whereas its stability to denaturation by heat and urea was markedly reduced. More importantly, upon renaturation of the urea-denatured mutant protein, a folding intermediate accumulated, having the characteristics of a molten globule. It had no tertiary structure, as shown by near UV circular dichroism, whereas the secondary structure was substantially recovered. The hydrophobic probe 8-anilino-1-naphthalene sulfonate bound to the intermediate species with an increase in fluorescence intensity and a blue shift. The hydrodynamic size was between that expected for a folded and an unfolded monomer. Finally, electrophoresis in a transverse urea gradient displayed no renaturation curve, and the protein showed the tendency to aggregate at the lowest urea concentrations. The existence of a molten globule folding intermediates resulting from an altered folding in the mutated protein might be related to the aggressiveness of neuroblastomas.


Evidence for Association Between Low Frequency Variants in CHRNA6/CHRNB3 and Antisocial Drug Dependence.
Common SNPs in nicotinic acetylcholine receptor genes (CHRN genes) have been associated with drug behaviors and personality traits, but the influence of rare genetic variants is not well characterized. The goal of this project was to identify novel rare variants in CHRN genes in the Center for Antisocial Drug Dependence (CADD) and Genetics of Antisocial Drug Dependence (GADD) samples and to determine if low frequency variants are associated with antisocial drug dependence. Two samples of 114 and 200 individuals were selected using a case/control design including the tails of the phenotypic distribution of antisocial drug dependence. The capture, sequencing, and analysis of all variants in 16 CHRN genes (CHRNA1-7, 9, 10, CHRNB1-4, CHRND, CHRNG, CHRNE) were performed independently for each subject in each sample. Sequencing reads were aligned to the human reference sequence using BWA prior to variant calling with the Genome Analysis ToolKit (GATK). Low frequency variants (minor allele frequency < 0.05) were analyzed using SKAT-O and C-alpha to examine the distribution of rare variants among cases and controls. In our larger sample, the region containing the CHRNA6/CHRNB3 gene cluster was significantly associated with disease status using both SKAT-O and C-alpha (unadjusted p values <0.05). More low frequency variants in the CHRNA6/CHRNB3 gene region were observed in cases compared to controls. These data support a role for genetic variants in CHRN genes and antisocial drug behaviors.


Characterization of the bovine tenascin-X.
The primary structure of flexilin, an extracellular matrix glycoprotein previously identified in bovine tissues (Lethias, C., Descollonges, Y., Boutillon, M.-M., and Garrone, R. (1996) Matrix Biol. 15, 11-19) was determined by cDNA cloning. The deduced amino acid sequence (4135 residues) reveals that this protein is composed of a succession of peptide motifs characteristic of the tenascin family: an amino-terminal domain containing cysteine residues and heptads of hydrophobic amino acids, 18.5 epidermal growth factor-like repeats, 30 fibronectin type III-like (FNIII) domains, and a carboxyl-terminal fibrinogen-like motif. Sequence analysis indicated that this protein is the bovine orthologue of human tenascin-X. By rotary shadowing, bovine tenascin-X was identified as monomers with a flexible aspect, which are ended by a globule. More FNIII motifs were characterized in the bovine protein than in human tenascin-X. The main difference between the human and bovine tenascin-X is found in the arrangement of the three classes of highly similar FNIII repeat types in the central region of tenascin-X. The bovine FNIII motif b10 exhibits an RGD putative cell attachment site. The functional role of this sequence is corroborated by cell adhesion on purified tenascin-X, which is inhibited by RGD peptides. Moreover, we demonstrate that this RGD site is conserved at the same location in the human molecule.


p53 expression in correlation to clinical outcome in patients with renal cell carcinoma.
The aim of this study was to evaluate the role of p53 as prognostic factor in renal cell carcinoma (RCC) and its relation to clinicopathological factors. The nuclear accumulation of p53 protein was determined by immunohistochemical analysis in RCC specimens from 90 patients and was correlated with clinical stage, grade, DNA ploidy, S-phase fraction and cancer-specific survival. p53 overexpression was observed in 17 of 90 (19%) tumours. There was a significant correlation to stage (p = 0.016) and grade (p = 0.020) but not to DNA ploidy or S-phase. Patients with high p53 immunoreactivity had shorter cancer-specific survival (p = 0.003) than those with normal p53 protein expression. This difference was found in papillary and chromophobe tumour types (p < 0.0001) but not in conventional RCC. In patients with RCC, significant correlations between p53 protein expression and tumour stage, grade and survival time were observed. For patients with chromophobe and papillary tumour types, but not in conventional RCC, p53 immunoreactivity gave prognostic information, suggesting that the prognostic differences in p53 immunoreactivity might be due to disparate genetic abnormalities in the different RCC types.


BACITRACIN: A NEW ANTIBIOTIC PRODUCED BY A MEMBER OF THE B. SUBTILIS GROUP.
A new antibiotic "bacitracin" has been recovered from a strain of the B. subtilis group of organisms. It is neutral, water-soluble, non-toxic and relatively heat stable. In vitro it is active chiefly against Grampositive organisms, but the gonococcus and meningococcus are susceptible to its action. It is also active in vivo against experimentally produced hemolytic streptococcal infections in mice and gas gangrene infections in guinea pigs. Clinical use in hemolytic streptococcal and staphylococcal infections in man have given encouraging results.


Pulmonary artery sarcomas. A review and report of a case.
We examined a case of primary pulmonary artery sarcoma and reviewed 45 cases that we found previously reported in the English literature, with particular attention given to the pathologic features. These sarcomas involve the pulmonary arterial trunk, left and right main pulmonary arteries, pulmonary valve, and right ventricular outflow tract. They have prominent intravascular growth along the arterial intima. Lung involvement commonly occurs by direct extension through the pulmonary vasculature or by distant metastases. These sarcomas contain a variety of heterologous components and areas of bone, cartilage, and fibrous tissue may be remarkably well-differentiated. Management of these sarcomas will be aided by an understanding of their biologic behavior and pathologic features.


The importance of quantitative trait differentiation in restoration: landscape heterogeneity and functional traits inform seed transfer guidelines.
For widely distributed species, understanding the scale over which genetic variation correlates to landscape structure and composition is critical. Particularly within the context of restoration, the evolution of genetic differences may impact success if seeds are maladapted to the restoration environment. In this study, we used Geum triflorum to quantify the scale over which genetic differences for quantitative traits important to adaptation have evolved, comparing the proportion of variance attributed to broad regional- and local population-level effects. Geum triflorum is a widely distributed species spanning a range of environments, including alvar and prairie habitats, which have extreme regional differences in soil-moisture availability. Alvar habitats are regions of thin soil over limestone that experience substantial seasonal variation in water availability, from flooding to desiccation annually. This contrasts with prairie habitats, whose deeper soils mitigate irregular flood-desiccation cycles. Using a common garden experiment, we evaluated 15 traits broadly grouped into three trait classes: resource allocation, stomatal characteristics, and leaf morphological traits for individuals sourced from prairie and alvar environments. We quantified the proportion of trait variance explained by regional- and population-scale effects and compared the proportion of regional- and population-trait variances explained across trait classes. Significant regional differentiation was observed for the majority of quantitative traits; however, population-scale effects were equal or greater than regional effects, suggesting that important genetic differences may have evolved across the finer population scale. Stomatal and resource allocation trait classes exhibited substantial regional differentiation relative to morphological traits, which may indicate increased strength of selection for stomatal and resource allocation traits relative to morphological traits. These patterns point towards the value in considering the scale over which genetic differences may have evolved for widely distributed species and identify different functional trait classes that may be valuable in establishing seed transfer guidelines.


Comparison of two vertical condensation obturation techniques: Touch 'n Heat modified and System B.
The aims of this study were firstly to compare the area of canal occupied by gutta-percha, sealer or voids using the System B heating device with that obtained by a modified vertical condensation technique using the Touch 'n Heat: and secondly to compare the temperature changes at the root canal wall and external root surface during obturation with the above techniques. Forty-five resin blocks, each with a standardized, simulated, prepared main root canal and five lateral canals, were assigned to three equal experimental groups. The canals were obturated using either the System B technique at two different temperature settings, or vertical condensation with a Touch 'n Heat instrument as the heat source. A heat transfer model was used to simultaneously record internal and external root surface temperature elevations during obturation by the three techniques. Data were analysed using unpaired Student's t-test and Mann-Whitney U-test. Both obturation techniques produced root fillings consisting of over 90% gutta-percha at most levels, although the percentages of sealer and voids 2-3 mm from the working length following System B obturation were higher than those found following modified vertical condensation. Modified vertical condensation resulted in more gutta-percha in lateral canals. Obturation was accomplished more quickly using the System B, and temperature elevations produced during obturation with the System B were significantly less (P < 0.001) than with vertical condensation. An elevation of external root surface temperature by more than 10 degrees C occurred during vertical condensation. The results suggest that the System B may produce an acceptable obturation and that the use of a Touch 'n Heat source during vertical condensation may result in damage to the periodontium.