Expression and localization of immunoreactive-sialomucin complex (Muc4) in salivary glands.
Sialomucin Complex (SMC; Muc4) is a heterodimeric glycoprotein consisting of two subunits, the mucin component ASGP-1 and the transmembrane subunit ASGP-2. Northern blot and immunoblot analyses demonstrated the presence of SMC/Muc4 in submaxillary, sublingual and parotid salivary glands of the rat. Immunocytochemical staining of SMC using monoclonal antisera raised against ASGP-2 and glycosylated ASGP-1 on paraffin-embedded sections of parotid, submaxillary and sublingual tissues was performed to examine the localization of the mucin in the major rat salivary glands. Histological and immunocytochemical staining of cell markers showed that the salivary glands consisted of varying numbers of serous and mucous acini which are drained by ducts. Parotid glands were composed almost entirely of serous acini, sublingual glands were mainly mucous in composition and a mixture of serous and mucous acini were present in submaxillary glands. Since immunoreactive (ir)-SMC was specifically localized to the serous cells, staining was most abundant in parotid glands, intermediate levels in submaxillary glands and least in sublingual glands. Ir-SMC in sublingual glands was localized to caps of cells around mucous acini, known as serous demilunes, which are also present in submaxillary glands. Immunocytochemical staining of SMC in human parotid glands was localized to epithelial cells of serous acini and ducts. However, the staining pattern of epithelial cells was heterogeneous, with ir-SMC present in some acinar and ductal epithelial cells but not in others. This report provides a map of normal ir-SMC/Muc4 distribution in parotid, submaxillary and sublingual glands which can be used for the study of SMC/Muc4 expression in salivary gland tumors.


Redefining High-Volume Gastric Cancer Centers: The Impact of Operative Volume on Surgical Outcomes.
Performance of technically complex surgery at high-volume (HV) centers is associated with improved outcomes. The aim of this study was to assess whether hospital gastrectomy volume is associated with surgical outcomes, and what threshold of case volume meaningfully impacts surgical outcomes. We conducted a retrospective review of adult NCDB patients with gastric adenocarcinoma undergoing gastrectomy between 2004 and 2015. A multivariable Cox proportional hazards model with restricted cubic splines was used to examine the association of annual hospital gastrectomy volume and overall survival. Bootstrap simulation was used to estimate the cut-point corresponding to maximum change in log hazard ratio. Hospitals were divided into HV (≥ 17 cases/year) and low-volume (LV; < 17 cases/year) groups. We examined the relationship between volume groups and adequate nodal examination, R0 resection, unplanned readmission, and 30- and 90-day mortality. Our cohort consisted of 29,559 patients (7.8% treated at an HV center). Treatment at an HV center was associated with an increased likelihood of adequate nodal examination [odds ratio (OR) 2.12, 95% confidence interval (CI) 1.94-2.32] and R0 resection among patients with cardia tumors (OR 1.42, 95% CI 1.07-1.88). Patients treated at HV centers had decreased 30- and 90-day postoperative mortality, which was more pronounced in those undergoing total gastrectomy. Treatment at an HV gastrectomy center is associated with improved surgical outcomes. Our study identified 17 cases/year as a clinically meaningful distinction between HV and LV centers. This definition of an HV center should be considered when evaluating regionalization of gastric cancer care to improve patient outcomes.


Long-chain n-3 fatty acids specifically affect rat coagulation factors dependent on vitamin K: relation to peroxidative stress.
Fatty acids of marine origin have been shown to affect blood coagulation in the rat. In an attempt to gain insight into the mechanisms of this phenomenon, we studied the effects of dietary linseed and fish oils on the liver antioxidant status and plasma coagulation parameters in rats on a time-course basis. Dietary enrichment in eicosapentaenoic and docosahexaenoic acids resulted in strong hypocoagulation after only 1 week and a concomitant increase in liver lipid peroxidation and tocopherolquinone content. Enrichment in linolenic acid induced similar increases in lipid peroxidation and tocopherol catabolism but negligible alteration of coagulation. A significant correlation between plasma factor II coagulant activity and liver tocopherolquinone was found in fish oil- but not in linseed oil-fed rats. Although ingestion of tocopherolquinone led to high levels of this compound in the liver, it had only marginal effects on coagulation factors. Thus, it seems unlikely that this vitamin E metabolite could be involved in the lowering of vitamin K-dependent clotting factors through inhibition of gamma-glutamylcarboxylase. Rather, our results indicate that the effects of the n-3 fatty acids of fish oil on vitamin K-dependent coagulation factors are specific and independent of liver tocopherolquinone levels.


Ecological context, concentrated disadvantage, and youth reoffending: identifying the social mechanisms in a sample of serious adolescent offenders.
Serious youthful offenders are presented with a number of significant challenges when trying to make a successful transition from adolescence to adulthood. One of the biggest obstacles for these youth to overcome concerns their ability to desist from further antisocial behavior, and although an emerging body of research has documented important risk and protective factors associated with desistance, the importance of the neighborhoods within which these youth reside has been understudied. Guided by the larger neighborhood effects on crime literature, the current study examines the direct and indirect effects of concentrated disadvantage on youth reoffending among a sample of highly mobile, serious youthful offenders. We use data from Pathways to Desistance, a longitudinal study of serious youthful offenders (N = 1,354; 13.6% female; 41.4% African American, 33.5% Hispanic, 20.2% White), matched up with 2000 Census data on neighborhood conditions for youth's main residence location during waves 7 and 8 of the study. These waves represent the time period in which youth are navigating the transition to adulthood (aged 18-22; average age = 20). We estimate structural equation models to determine direct effects of concentrated disadvantage on youth reoffending and also to examine the possible indirect effects working through individual-level mechanisms as specified by theoretical perspectives including social control (e.g., unsupervised peer activities), strain (e.g., exposure to violence), and learning (e.g., exposure to antisocial peers). Additionally, we estimate models that take into account the impact that a change in neighborhood conditions may have on the behavior of youth who move to new residences during the study period. Our results show that concentrated disadvantage is indirectly associated with youth reoffending primarily through its association with exposure to deviant peers. Taking into account youth mobility during the study period produced an additional indirect pathway by which concentrated disadvantage is associated with goal blockage (i.e., the gap between belief in conventional goals and perceived potential to reach those goals), which was then associated with exposure to deviant peers and indirectly, reoffending behavior. We conclude that the neighborhood effects literature offers a promising framework for continued research on understanding the successful transition to adulthood by serious youthful offenders.


Beyond the beat: A pioneering investigation into exercise modalities for alleviating diabetic cardiomyopathy and enhancing cardiac health.
Patients with preexisting cardiovascular disease or those at high risk for developing the condition are often offered exercise as a form of therapy. Patients with cancer who are at an increased risk for cardiovascular issues are increasingly encouraged to participate in exercise-based, interdisciplinary programs due to the positive correlation between these interventions and clinical outcomes following myocardial infarction. Diabetic cardiomyopathy (DC) is a cardiac disorder that arises due to disruptions in the homeostasis of individuals with diabetes. One of the primary reasons for mortality in individuals with diabetes is the presence of cardiac structural damage and functional abnormalities, which are the primary pathological features of DC. The aetiology of dilated cardiomyopathy is multifaceted and encompasses a range of processes, including metabolic abnormalities, impaired mitochondrial function, dysregulation of calcium ion homeostasis, excessive cardiomyocyte death, and fibrosis. In recent years, many empirical investigations have demonstrated that exercise training substantially impacts the prevention and management of diabetes. Exercise has been found to positively impact the recovery of diabetes and improve several metabolic problem characteristics associated with DC. One potential benefit of exercise is its ability to increase systolic activity, which can enhance cardiometabolic and facilitate the repair of structural damage to the heart caused by DC, leading to a direct improvement in cardiac health. In contrast, exercise has the potential to indirectly mitigate the pathological progression of DC through its ability to decrease circulating levels of sugar and fat while concurrently enhancing insulin sensitivity. A more comprehensive understanding of the molecular mechanism via exercise facilitates the restoration of DC disease must be understood. Our goal in this review was to provide helpful information and clues for developing new therapeutic techniques for motion alleviation DC by examining the molecular mechanisms involved.


Signal-dependent turnover of the bacterial flagellar switch protein FliM.
Most biological processes are performed by multiprotein complexes. Traditionally described as static entities, evidence is now emerging that their components can be highly dynamic, exchanging constantly with cellular pools. The bacterial flagellar motor contains approximately 13 different proteins and provides an ideal system to study functional molecular complexes. It is powered by transmembrane ion flux through a ring of stator complexes that push on a central rotor. The Escherichia coli motor switches direction stochastically in response to binding of the response regulator CheY to the rotor switch component FliM. Much is known of the static motor structure, but we are just beginning to understand the dynamics of its individual components. Here we measure the stoichiometry and turnover of FliM in functioning flagellar motors, by using high-resolution fluorescence microscopy of E. coli expressing genomically encoded YPet derivatives of FliM at physiological levels. We show that the approximately 30 FliM molecules per motor exist in two discrete populations, one tightly associated with the motor and the other undergoing stochastic turnover. This turnover of FliM molecules depends on the presence of active CheY, suggesting a potential role in the process of motor switching. In many ways the bacterial flagellar motor is as an archetype macromolecular assembly, and our results may have further implications for the functional relevance of protein turnover in other large molecular complexes.


Mutation site and context dependent effects of ESR1 mutation in genome-edited breast cancer cell models.
Mutations in the estrogen receptor alpha (ERα) 1 gene (ESR1) are frequently detected in ER+ metastatic breast cancer, and there is increasing evidence that these mutations confer endocrine resistance in breast cancer patients with advanced disease. However, their functional role is not well-understood, at least in part due to a lack of ESR1 mutant models. Here, we describe the generation and characterization of genome-edited T47D and MCF7 breast cancer cell lines with the two most common ESR1 mutations, Y537S and D538G. Genome editing was performed using CRISPR and adeno-associated virus (AAV) technologies to knock-in ESR1 mutations into T47D and MCF7 cell lines, respectively. Various techniques were utilized to assess the activity of mutant ER, including transactivation, growth and chromatin-immunoprecipitation (ChIP) assays. The level of endocrine resistance was tested in mutant cells using a number of selective estrogen receptor modulators (SERMs) and degraders (SERDs). RNA sequencing (RNA-seq) was employed to study gene targets of mutant ER. Cells with ESR1 mutations displayed ligand-independent ER activity, and were resistant to several SERMs and SERDs, with cell line and mutation-specific differences with respect to magnitude of effect. The SERD AZ9496 showed increased efficacy compared to other drugs tested. Wild-type and mutant cell co-cultures demonstrated a unique evolution of mutant cells under estrogen deprivation and tamoxifen treatment. Transcriptome analysis confirmed ligand-independent regulation of ERα target genes by mutant ERα, but also identified novel target genes, some of which are involved in metastasis-associated phenotypes. Despite significant overlap in the ligand-independent genes between Y537S and D538G, the number of mutant ERα-target genes shared between the two cell lines was limited, suggesting context-dependent activity of the mutant receptor. Some genes and phenotypes were unique to one mutation within a given cell line, suggesting a mutation-specific effect. Taken together, ESR1 mutations in genome-edited breast cancer cell lines confer ligand-independent growth and endocrine resistance. These biologically relevant models can be used for further mechanistic and translational studies, including context-specific and mutation site-specific analysis of the ESR1 mutations.


Binocular coordination during reading and non-reading tasks.
The goal of this review is to evaluate the literature on binocular coordination during reading and non-reading tasks in adult, child, and dyslexic populations. The review begins with a description of the basic characteristics of eye movements during reading. Then, reading and non-reading studies investigating binocular coordination are evaluated. Areas of future research in the field are identified and discussed. Finally, some general conclusions are made regarding binocular coordination. The review demonstrates that findings from traditionally independent areas of research are largely consistent and complementary. Throughout the review, theoretical and methodological commonalities are identified and clarified in order to advance current understanding of this fundamental aspect of human visual processing.


Surgical treatment of pelvic sarcoma in children: outcomes for twenty six patients.
Pelvic sarcoma is uncommon in children and challenging to treat. This study examined different surgical approaches to treat pelvic sarcoma with the aim of assessing the oncologic, and functional outcomes. We retrospectively examined the medical records of patients younger than 21 years of age who underwent surgery for pelvic sarcoma at our institution from 1992 to 2010. The functional status of the patients was examined after a minimum follow-up of two years. Twenty-six patients were included in the analysis. Nineteen (73%) patients were male and seven (27%) were female. Mean age at presentation was 12.0 ± 3.9 years. Nineteen patients had Ewing sarcoma (73%), five had osteosarcoma (19%), one had chondrosarcoma (4%) and one had rhabdomyosarcoma (4%). Iliac wing resection with no reconstruction was done in three patients. Reconstruction with free fibular graft A-frame was performed in four patients, saddle endoproshtesis in five patients, iliac autoclave in one patient, and internal hemipelvectomy in nine patients. Hindquarter amputation was performed in five patients. Median follow-up was 4.6 years (range, 2.6-16 years). Nineteen patients were alive (73%); of those, 13 were known to be without disease, three were with disease and three did not have known tumor status. Six patients were reported deceased, three had osteosarcoma and three had Ewing sarcoma. Function was assessed in 17 patients; 64% were asymptomatic and ambulatory and 36% were symptomatic and ambulatory. Salvage reconstruction for pelvic sarcoma can be performed through various procedures on the extent of necessary bony resection. Survival rate and functional outcomes were promising in the performed study.


Virulence of a polymicrobic complex, Treponema denticola and Porphyromonas gingivalis, in a murine model.
The effect of a polymicrobic infection employing Treponema denticola and Porphyromonas gingivalis in the murine lesion model was used to determine the synergistic virulence of these two periodontopathic bacteria. At high doses of P. gingivalis W50, addition of T. denticola in the infection mixture had no effect on the formation and size of the spreading lesion caused by this microorganism. However, at low P. gingivalis challenge doses, T. denticola significantly enhanced the virulence of P. gingivalis compared with monoinfection of this microorganism. A potential role of the trypsin-like protease enzyme activity of P. gingivalis in this synergistic virulence was tested using P. gingivalis mutants deficient (i.e., BEI) or devoid (i.e., NG4B19) of this protease activity. These findings demonstrated that T. denticola-P. gingivalis complexes exhibit enhanced virulence in this model and that even using a polymicrobic challenge infection, the trypsin-like protease activity was important to P. gingivalis virulence expression.


Physicochemical strategies for inhibition of amyloid fibril formation: an overview of recent advances.
Protein aggregation and amyloid fibrillation can lead to several serious human diseases and protein drug ineffectiveness. The complexity and dynamics of protein folding present unique challenges for elucidating the molecular mechanisms involved in protein aggregation and designing effective amyloid inhibitors. Continuous development of creative approaches to identify an ultimate solution for controlling protein aggregation in biopharmaceuticals and clinical pathology is clearly required. This review describes and discusses the most recent advances on the physicochemical strategies for inhibiting protein aggregation and amyloid fibrillation, with emphasis on giving a brief overview of creative approaches and chemistries used. Physical strategies for inhibiting amyloid fibril formation, including high hydrostatic pressure, low temperature, and laser irradiation, are critically evaluated. Recent advances in chemical strategies including small molecules, metal chelators, and nanomaterials, as well as in the use of biomolecules (peptide, protein, nucleic acid, and saccharide) as amyloid inhibitors, are also highlighted.


What does the absence of Helicobacter pylori from the antrum imply?
Helicobacter pylori is absent in the antrum of approximately 5, 25, and 35% of patients with duodenal ulcer, gastric ulcer, and non-ulcer dyspepsia respectively and in the majority of asymptomatic healthy subjects from the West. The absence of H. pylori from the antrum could result from failure of exposure to (Group A) or colonization by (Group B) or temporary (Group C) or permanent (Group D) clearance of H. pylori, after initial colonization. Marked differences in the incidence of absence of H. pylori in antrum of different age group of control subjects from the West and the developing nation (such as India) are discussed.


On-the-fly determination of active region centers in adaptive-partitioning QM/MM.
Quantum mechanics/molecular mechanics (QM/MM) methods are widely used in molecular dynamics (MD) simulations of large systems. By partitioning the system into active and environmental regions and treating them with different levels of theory, QM/MM methods achieve accuracy and efficiency at the same time. Adaptive-partitioning (AP) QM/MM allows the partition of the system to change during the MD simulation, making it possible to simulate processes in which the active and environmental regions exchange atoms or molecules, such as processes in solutions or solids. AP-QM/MM methods usually partition the system according to distances to centers of active regions. For energy-conserving AP-QM/MM methods, these centers are chosen beforehand and remain fixed during the MD simulation, making it difficult to simulate processes in which active regions may occur or vanish. In this paper, I develop an adaptive-center (AC) method that allows on-the-fly determination of the centers of active regions according to any geometrical criterion or any criterion dependent on the potential energy. The AC method is compatible with all existing energy-conserving AP-QM/MM methods, and the resulting potential energy surface is smooth. The application of the AC method is demonstrated with two examples in solid systems.


Structure and biosynthesis of cytoplasmic and secreted variants of gelsolin.
Gelsolin is an actin-fragmenting cytoplasmic protein. A functionally similar protein has also been identified in plasma. We have compared the structure of the cytoplasmic and plasma forms of gelsolin and examined their biosynthetic relationships. Plasma gelsolin is larger than cytoplasmic gelsolin (Mr 93,000 versus 90,000, respectively) and is more positively charged. Partial amino acid sequencing analyses show that the two gelsolins share a common 29 amino acid sequence which lies at the NH2-terminal end of cytoplasmic gelsolin and spans residues 26-55 of plasma gelsolin. Compared with cytoplasmic gelsolin, plasma gelsolin contains an additional peptide of 25 amino acids at its NH2 terminus. The human hepatoma-derived cell line, HepG2, synthesizes both the 90-kDa and the 93-kDa gelsolins but secretes only the 93-kDa form. Pulse-chase experiments demonstrate that the rate of disappearance of the 93-kDa gelsolin from the cells corresponds with the rate of appearance of the 93-kDa gelsolin in the medium, whereas the rate of disappearance of the 90-kDa gelsolin is independent of and slower than that of the secreted plasma protein. We conclude that cytoplasmic and plasma gelsolins are structurally similar but not identical, that after synthesis these proteins are processed independently, and that the fate of each is distinct.


Formation of a bi-rhodium boron tube Rh2B18 and its great CO2 capture ability.
The geometries, bonding and abilities for CO2 capture of the doubly rhodium-doped boron cluster Rh2B18 are presented. DFT calculations using the TPSSh functional show that the doubly doped Rh2B18 is stable in a high symmetry shape (D9d) in which two Rh dopants are vertically and oppositely coordinated to nonagonal faces of a (2 × 9) double ring B18 tube, having the form of a teetotum toy. Bonding of the tube is analysed using different schemes for partition of total electron density (EDI_D, AIM). The high thermodynamic stability of Rh2B18 can be rationalized in terms of its electron distribution in which both Rh atoms share delocalized bonds with B atoms. Molecular dynamic simulations also confirm its stability within a range of temperatures. Exploration of the interaction of gas phase CO2 molecules with Rh2B18 suggests that this tubular structure has a great ability for capture and transformation of carbon dioxide. Formation of two new strong bonds of CO2 with the B-B edges of Rh2B18 favour its capture through a dissociative adsorption mechanism in which a nearly spontaneous dissociation of CO2 into CO and BO groups follows its attachment.


Vitamin E for treating children's scars. Does it help reduce scarring?
A few of my patients have been told to use vitamin E cream after surgery or repair of a laceration. What is the evidence for this suggestion, and is this treatment suitable for all patients? Vitamin E is the main lipid-soluble antioxidant in the skin. Several anecdotal reports have suggested that topical use of vitamin E cream can reduce scar formation. Current evidence from the literature, however, does not support that proposition. In fact, studies report some adverse effects with use of vitamin E. Further research is needed before application of vitamin E cream becomes the standard of care.


Color stability of recent composite resins.
The objective of the study was to evaluate the color stability of 8 recently developed resin composites when exposed to various staining agents. Six disc-shaped specimens made out of 8 resin composite materials were immersed in artificial saliva, coffee, coke, tea, orange juice and red wine. The initial color (T0) of the 288 specimens was assessed by a calibrated reflectance spectrophotometer (SpectroShade, MHT) over a black as well as a white background. All specimens were kept in an incubator (INP-500, Memmert GmbH) at 37 °C for 4 weeks. Colorant solutions were changed every 7 days to avoid bacteria or yeast contamination. After 4 weeks of storage in the colorants (T1), spectrophotometric measurements were repeated and ΔE00 was calculated. When analyzed over a white background, the mean ΔE00 values varied from 1.5 (Venus Diamond) to 52.4 (Miris 2). When analyzed over a black background, the mean ΔE00 values varied from 0.8 (Venus Diamond) to 40.0 (ELS). Significant differences were detected when comparing values at T0 and T1 (ΔE00) for both backgrounds (Anova and Fisher's LSD post hoc test). Resin composite materials do behave in a significantly different way (p value < 0.01) when exposed to various staining solutions.


Paradoxic effects of metformin on endothelial cells and angiogenesis.
The biguanide metformin is used in type 2 diabetes management and has gained significant attention as a potential cancer preventive agent. Angioprevention represents a mechanism of chemoprevention, yet conflicting data concerning the antiangiogenic action of metformin have emerged. Here, we clarify some of the contradictory effects of metformin on endothelial cells and angiogenesis, using in vitro and in vivo assays combined with transcriptomic and protein array approaches. Metformin inhibits formation of capillary-like networks by endothelial cells; this effect is partially dependent on the energy sensor adenosine-monophosphate-activated protein kinase (AMPK) as shown by small interfering RNA knockdown. Gene expression profiling of human umbilical vein endothelial cells revealed a paradoxical modulation of several angiogenesis-associated genes and proteins by metformin, with short-term induction of vascular endothelial growth factor (VEGF), cyclooxygenase 2 and CXC chemokine receptor 4 at the messenger RNA level and downregulation of ADAMTS1. Antibody array analysis shows an essentially opposite regulation of numerous angiogenesis-associated proteins in endothelial and breast cancer cells including interleukin-8, angiogenin and TIMP-1, as well as selective regulation of angiopioetin-1, -2, endoglin and others. Endothelial cell production of the cytochrome P450 member CYP1B1 is upregulated by tumor cell supernatants in an AMPK-dependent manner, metformin blocks this effect. Metformin inhibits VEGF-dependent activation of extracellular signal-regulated kinase 1/2, and the inhibition of AMPK activity abrogates this event. Metformin hinders angiogenesis in matrigel pellets in vivo, prevents the microvessel density increase observed in obese mice on a high-fat diet, downregulating the number of white adipose tissue endothelial precursor cells. Our data show that metformin has an antiangiogenic activity in vitro and in vivo associated with a contradictory short-term enhancement of pro-angiogenic mediators, as well as with a differential regulation in endothelial and breast cancer cells.


Thyrotrophin estimation in diagnosis and treatment of childhood thyroid disorders.
Serum thyrotrophin (TSH) was estimated by double-antibody radioimmunoassay in 200 children aged 2 months to 16 years with normal thyroid function. There was no apparent variation in TSH with age or sex and only 4 children had TSH levels greater than 5 muU/ml. High TSH values were obtained in 9 children with primary hypothyroidism, in 3 children with thyroiditis, and in one girl with a lingual thyroid. Moderately raised TSH was found in 3 girls with thyroiditis, 2 brothers with goitres due to enzyme defect, and a girl with an ectopic thyroid. In one girl with a defect of iodine organification and in 3 boys with thyroxine binding globulin deficiency the TSH levels were normal despite very low serum thyroxine values. Serum TSH was also estimated in 20 children during treatment for primary hypothyroidism. 3 of these children showed slightly raised TSH levels despite apparently adequate replacement therapy with L-thyroxine. One girl showed a very high TSH level 3 weeks after treatment had been temporarily withdrawn.


Protective Effect of Alpha-Linolenic Acid on Human Oral Squamous Cell Carcinoma Metastasis and Apoptotic Cell Death.
Oral cancer ranks sixth among Taiwan's top 10 cancers and most patients with poor prognosis acquire metastases. The essential fatty acid alpha-linolenic acid (ALA) has been found to diminish many cancer properties. However, the anti-cancer activity of ALA in oral cancer has yet to be determined. We examined the mechanisms underlying ALA inhibition of metastasis and induction of apoptotic cell death in oral squamous cell carcinoma (OSCC). Migration and invasion assays confirmed the cancer cells' EMT capabilities, whereas flow cytometry and Western blotting identified molecular pathways in OSCC. ALA dramatically reduced cell growth in a concentration-dependent manner according to the findings. Low concentrations of ALA (100 or 200 μM) inhibit colony formation, the expression of Twist and EMT-related proteins, the expression of MMP2/-9 proteins, and enzyme activity, as well as cell migration and invasion. Treatment with high concentrations of ALA (200 or 400 μM) greatly increases JNK phosphorylation and c-jun nuclear accumulation and then upregulates the FasL/caspase8/caspase3 and Bid/cytochrome c/caspase9/caspase3 pathways, leading to cell death. Low concentrations of ALA inhibit SAS and GNM cell migration and invasion by suppressing Twist and downregulating EMT-related proteins or by decreasing the protein expression and enzyme activity of MMP-2/-9, whereas high concentrations of ALA promote apoptosis by activating the JNK/FasL/caspase 8/caspase 3-extrinsic pathway and the Bid/cytochrome c/caspase 9 pathway. ALA demonstrates potential as a treatment for OSCC patients.


[The effect of comorbid headaches in patients with epilepsy on clinical pathomorphosis of the disease].
Of 404 adult patients with epilepsy (E) with a period of follow-up from 5 to 26 years (mean 8.5 +/- 3.0) a group of 168 people was distinguished by presence of comorbid headaches (HAs) of various types: by origin--primary and secondary, by relation to occurrence of epileptic seizures--inter- and periictal. In general, the presence of comorbid HA did not affect the prognosis for remission of epileptic seizures in patients of all etiologic groups (idiopathic, cryptogenic, and symptomatic). Primary HAs were mainly observed in cryptogenic and idiopathic E, and secondary--in symptomatic E. It is in the group of patients with symptomatic epilepsy and HAs the clinical pathomorphosis depending on the etiology of the disease has been marked. HA in patients with vascular E haven't worsen the prognosis of the disease: 86.7% of patients were in remission, indicating the positive clinical pathomorphosis of this form of E. At the same time, a negative long-term clinical pathomorphosis of symptomatic E was observed in comorbidity of HA with posttraumatic E, with mesial temporal sclerosis and with E due to the brain space-occupying lesions, or the consequences of their removal. In these cases the patients developed more often drug-resistance. Pseudo-resistance was observed in patients with E and HA in multiple sclerosis and in alcoholic E.


Combined Effects of Nanoroughness and Ions Produced by Electrodeposition of Mesoporous Bioglass Nanoparticle for Bone Regeneration.
Providing appropriate biophysical and biochemical cues to the interface is a facile strategy to enhance the osteogenic ability of metallic implants. Here we exploited this through the incorporation of mesoporous bioactive glass nanoparticles (MBGN) at a high content (1:1 by weight) to a biopolymer chitosan in the electrodeposition process of titanium. The MGBN/chitosan layer thickness, tunable by electrodeposition parameters, exhibited an accelerated ability of apatite mineral induction in a body simulating medium. Of note, the involvement of MBGN could generate nanoscale roughness in a unique range of 10-25 nm. Moreover, the layer showed a slowly releasing profile of ions (calcium and silicate) over weeks at therapeutically relevant doses. The ion-releasing nanotopological surface was demonstrated to alter the preosteoblasts responses in a way favorable for osteogenic differentiation. The combinatory cues of nanotopology (25 nm roughness) and ion release enabled highly accelerated cellular anchorage with somewhat limited spreading area at initial periods. The subsequent osteoblastic differentiation behaviors on the engineered surface, as examined up to 21 days, showed significantly enhanced alkaline phosphate activity and up-regulated expression of bone-associated genes (ALP, Col I, OPN, and OCN). These results indicate that the combinatory cues provided by nanotopology (25 nm roughness) and ions released from MBGN are highly effective in stimulating osteoblastic differentiation and suggest that the MBGN/chitosan may serve as a potential composition for bone implant coatings.


Carbon Double Coated Fe3O4@C@C Nanoparticles: Morphology Features, Magnetic Properties, Dye Adsorption.
This work is devoted to the study of magnetic Fe3O4 nanoparticles doubly coated with carbon. First, Fe3O4@C nanoparticles were synthesized by thermal decomposition. Then these synthesized nanoparticles, 20-30 nm in size were processed in a solution of glucose at 200 °C during 12 h, which led to an unexpected phenomenon-the nanoparticles self-assembled into large conglomerates of a regular shape of about 300 nm in size. The morphology and features of the magnetic properties of the obtained hybrid nanoparticles were characterized by transmission electron microscopy, differential thermo-gravimetric analysis, vibrating sample magnetometer, magnetic circular dichroism and Mössbauer spectroscopy. It was shown that the magnetic core of Fe3O4@C nanoparticles was nano-crystalline, corresponding to the Fe3O4 phase. The Fe3O4@C@C nanoparticles presumably contain Fe3O4 phase (80%) with admixture of maghemite (20%), the thickness of the carbon shell in the first case was of about 2-4 nm. The formation of very large nanoparticle conglomerates with a linear size up to 300 nm and of the same regular shape is a remarkable peculiarity of the Fe3O4@C@C nanoparticles. Adsorption of organic dyes from water by the studied nanoparticles was also studied. The best candidates for the removal of dyes were Fe3O4@C@C nanoparticles. The kinetic data showed that the adsorption processes were associated with the pseudo-second order mechanism for cationic dye methylene blue (MB) and anionic dye Congo red (CR). The equilibrium data were more consistent with the Langmuir isotherm and were perfectly described by the Langmuir-Freundlich model.


Built Environment Features and Cardiometabolic Mortality and Morbidity in Remote Indigenous Communities in the Northern Territory, Australia.
Indigenous Australians experience poorer health than non-Indigenous Australians, with cardiometabolic diseases (CMD) being the leading causes of morbidity and mortality. Built environmental (BE) features are known to shape cardiometabolic health in urban contexts, yet little research has assessed such relationships for remote-dwelling Indigenous Australians. This study assessed associations between BE features and CMD-related morbidity and mortality in a large sample of remote Indigenous Australian communities in the Northern Territory (NT). CMD-related morbidity and mortality data were extracted from NT government health databases for 120 remote Indigenous Australian communities for the period 1 January 2010 to 31 December 2015. BE features were extracted from Serviced Land Availability Programme (SLAP) maps. Associations were estimated using negative binomial regression analysis. Univariable analysis revealed protective effects on all-cause mortality for the BE features of Education, Health, Disused Buildings, and Oval, and on CMD-related emergency department admissions for the BE feature Accommodation. Incidence rate ratios (IRR's) were greater, however, for the BE features Infrastructure Transport and Infrastructure Shelter. Geographic Isolation was associated with elevated mortality-related IRR's. Multivariable regression did not yield consistent associations between BE features and CMD outcomes, other than negative relationships for Indigenous Location-level median age and Geographic Isolation. This study indicates that relationships between BE features and health outcomes in urban populations do not extend to remote Indigenous Australian communities. This may reflect an overwhelming impact of broader social inequity, limited correspondence of BE measures with remote-dwelling Indigenous contexts, or a 'tipping point' of collective BE influences affecting health more than singular BE features.


Dynamic Changes of Lymphocyte Subsets in the Course of COVID-19.
Although the pathophysiology of coronavirus disease 2019 (COVID-19) is not clearly defined, among the proposed mechanisms, immune system dysfunction is more likely than others. The aim of this study was to clarify the characteristics and clinical significance of dynamic changes of lymphocyte subsets in the course of COVID-19. In this prospective study, the levels of peripheral lymphocyte subsets including CD4+, CD8+, CD4+CD25+FOXP3+, CD38+, CD3+HLA-DR+, CD19+, CD20+, and CD16+CD56+ cells were measured by flow cytometry in 52 confirmed hospitalized patients with COVID-19 at the day of admission and after 7 days of care. Clinical response was defined as improvement in symptoms (fever, dyspnea, and cough as well as blood oxygen saturation), and patients who met these criteria after 1 week of admission were classified as early responders; others who survived and finally discharged from the hospital were classified as late responders and patients who died were categorized as nonresponders. Immunophenotyping of studied cell changes on the first day of admission and 7 days after treatment were compared. Besides, the correlation between cellular subset variation and clinical response and outcome were analyzed. Total counts of white blood cell, T cells, CD4+ T cells, CD8+ T cells, CD38+ lymphocytes, and CD3+HLA-DR+ lymphocytes were significantly increased in both early and late responders. No statistically significant difference was observed in CD4+/CD8+ ratio, B cells, FOXP3+Treg lymphocytes, and FOXP3 median fluorescence intensity among studied groups. According to the multivariate analysis, an increase in CD4+ T cells (p = 0.019), CD8+ T cells (p = 0.001), and administration of interferon (p < 0.001) were independent predictors of clinical response. We found an increasing trend in total T cells, T helpers, cytotoxic T cells, activated lymphocytes, and natural killer cells among responders. This trend was not statistically significant among nonresponders. The findings of this study may enhance our knowledge about the pathogenesis of COVID-19.


The role of gender in a large international OCD sample: A Report from the International College of Obsessive-Compulsive Spectrum Disorders (ICOCS) Network.
Obsessive-compulsive disorder (OCD) is characterized by a range of phenotypic expressions. Gender may be a relevant factor in mediating the disorder's heterogeneity. The aim of the present report was to explore a large multisite clinical sample of OCD patients, hypothesizing existing demographic, geographical and clinical differences between male and female patients with OCD. Socio-demographic and clinical variables of 491 adult OCD outpatients recruited in the International College of Obsessive-Compulsive Spectrum Disorders (ICOCS) network were investigated with a retrospective analysis on a previously gathered set of data from eleven countries worldwide. Patients were assessed through structured clinical interviews, the Yale- Brown Obsessive-Compulsive Scale (Y-BOCS), the Montgomery-Asberg Depression Rating Scale (MADRS) and the Self-rating Depression Scale (SDS). Among females, adult onset (>18 years old) was significantly over-represented (67% vs. 33%, p < 0.005), and females showed a significantly older age at illness onset compared with males (20.85 ± 10.76 vs. 17.71 ± 8.96 years, p < 0.005). Females also had a significantly lower education level than males (13.09 ± 4.02 vs. 13.98 ± 3.85 years; p < 0.05), a significantly higher rate of being married (50.8% vs. 33.5%; p < 0.001) and a higher rate of living with a partner (47.5% vs. 37.6%; p < 0.001) than males. Nonetheless, no significant gender differences emerged in terms of the severity of OCD symptoms nor in the severity of comorbid depressive symptoms. No predictive effect of gender was found for Y-BOCS, MADRS and SDS severity. Our findings showed significant differences between genders in OCD. A sexually dimorphic pattern of genetic susceptibility may have a crucial role to OCD clinical heterogeneity, potentially requiring different specific therapeutic strategies. Further research is warranted to validate gender as an important determinant of the heterogeneity in OCD.


The influence of alcohol consumed with a meal on endothelial function in healthy individuals.
Alcohol and polyphenols in wine and fruit juices have been strongly implicated in the favourable effects on of these beverages on vascular function. Despite a wealth of information on the metabolic and vascular effects of alcohol and polyphenols, the combined influences of these substances on vascular function, especially when consumed with food, is poorly understood. A study was designed to determine the effects of a phenolic-rich grape juice, with or without alcohol, on vascular endothelial function in the postprandial state. Ten subjects consumed a standard meal with a test drink on three separate occasions. On each occasion, the test drink accompanying the meal was either red grape juice, red grape juice plus alcohol (12% v/v), or water. Endothelial function was measured by flow mediated dilatation (FMD) prior to then 30 and 60 minutes after consuming the meal. Blood samples were taken for the determination of plasma glucose, triacylglycerol (TAG) and non esterified fatty acids (NEFA) at regular intervals. There was a significant effect of the three treatments (P = 0.0026) and time (P = 0.021) on percentage FMD. The meals with the grape juice and grape juice plus alcohol produced similar FMD responses but were both significantly greater than the meal with water. The concentration of plasma glucose, TAG and NEFA were similar after each treatment. Alcohol had no effect on vascular function in the early postprandial phase. These findings provide new evidence to support the potential benefit of non-alcoholic components within alcoholic beverages on vascular function in the fed state.


Terminal organization of the corticospinal projection from the arm/hand region of the rostral primary motor cortex (M1r or old M1) to the cervical enlargement (C5-T1) in rhesus monkey.
High-resolution anterograde tracers and stereology were used to study the terminal organization of the corticospinal projection (CSP) from the rostral portion of the primary motor cortex (M1r) to spinal levels C5-T1. Most of this projection (90%) terminated contralaterally within laminae V-IX, with the densest distribution in lamina VII. Moderate bouton numbers occurred in laminae VI, VIII, and IX with few in lamina V. Within lamina VII, labeling occurred over the distal-related dorsolateral subsectors and proximal-related ventromedial subsectors. Within motoneuron lamina IX, most terminations occurred in the proximal-related dorsomedial quadrant, followed by the distal-related dorsolateral quadrant. Segmentally, the contralateral lamina VII CSP gradually declined from C5-T1 but was consistently distributed at C5-C7 in lamina IX. The ipsilateral CSP ended in axial-related lamina VIII and adjacent ventromedial region of lamina VII. These findings demonstrate the M1r CSP influences distal and proximal/axial-related spinal targets. Thus, the M1r CSP represents a transitional CSP, positioned between the caudal M1 (M1c) CSP, which is 98% contralateral and optimally organized to mediate distal upper extremity movements (Morecraft et al., 2013), and dorsolateral premotor (LPMCd) CSP being 79% contralateral and optimally organized to mediate proximal/axial movements (Morecraft et al., 2019). This distal to proximal CSP gradient corresponds to the clinical deficits accompanying caudal to rostral motor cortex injury. The lamina IX CSP is considered in the light of anatomical and neurophysiological evidence which suggests M1c gives rise to the major proportion of the cortico-motoneuronal (CM) projection, while there is a limited M1r CM projection.


Genetic differentiation of a southern Africa tepary bean (Phaseolus acutifolius A Gray) germplasm collection using high-density DArTseq SNP markers.
Genetic resources of tepary bean (Phaseolus acutifolius A. Gray) germplasm collections are not well characterized due to a lack of dedicated genomic resources. There is a need to assemble genomic resources specific to tepary bean for germplasm characterization, heterotic grouping, and breeding. Therefore, the objectives of this study were to deduce the genetic groups in tepary bean germplasm collection using high-density Diversity Array Technology (DArT) based single nucleotide polymorphism (SNP) markers and select contrasting genotypes for breeding. Seventy-eight tepary bean accessions were genotyped using 10527 SNPs markers, and genetic parameters were estimated. Population structure was delineated using principal component and admixture analyses. A mean polymorphic information content (PIC) of 0.27 was recorded, indicating a relatively low genetic resolution of the developed SNPs markers. Low genetic variation (with a genetic distance [GD] = 0.32) existed in the assessed tepary bean germplasm collection. Population structure analysis identified five sub-populations through sparse non-negative matrix factorization (snmf) with high admixtures. Analysis of molecular variance indicated high genetic differentiation within populations (61.88%) and low between populations (38.12%), indicating high gene exchange. The five sub-populations exhibited variable fixation index (FST). The following genetically distant accessions were selected: Cluster 1:Tars-Tep 112, Tars-Tep 10, Tars-Tep 23, Tars-Tep-86, Tars-Tep-83, and Tars-Tep 85; Cluster 3: G40022, Tars-Tep-93, and Tars-Tep-100; Cluster 5: Zimbabwe landrace, G40017, G40143, and G40150. The distantly related and contrasting accessions are useful to initiate crosses to enhance genetic variation and for the selection of economic traits in tepary bean.


Site-directed mutants of RTP of Bacillus subtilis and the mechanism of replication fork arrest.
DNA replication fork arrest during the termination phase of chromosome replication in Bacillus subtilis is brought about by the replication terminator protein (RTP) bound to specific DNA terminator sequences (Ter sites) distributed throughout the terminus region. An attractive suggestion by others was that crucial to the functioning of the RTP-Ter complex is a specific interaction between RTP positioned on the DNA and the helicase associated with the approaching replication fork. In support of this was the behaviour of two site-directed mutants of RTP. They appeared to bind Ter DNA normally but were ineffective in fork arrest as ascertained by in vitro Escherichia coli DnaB helicase and replication assays. We describe here a system for assessing the fork-arrest behaviour of RTP mutants in a bona fide in vivo assay in B. subtilis. One of the previously studied mutants, RTP.Y33N, was non-functional in fork arrest in vivo, as predicted. But through extensive analyses, this RTP mutant was shown to be severely defective in binding to Ter DNA, contrary to expectation. Taken in conjunction with recent findings on the other mutant (RTP.E30K), it is concluded that there is as yet no substantive evidence from the behaviour of RTP mutants to support the RTP-helicase interaction model for fork arrest. In an extension of the present work on RTP.Y33N, we determined the dissociation rates of complexes formed by wild-type (wt) RTP and another RTP mutant with various terminator sequences. The functional wtRTP-TerI complex was quite stable (half-life of 182 minutes), reminiscent of the great stability of the E. coli Tus-Ter complex. More significant were the exceptional stabilities of complexes comprising wtRTP and an RTP double-mutant (E39K.R42Q) bound to some particular terminator sequences. From the measurement of in vivo fork-arrest activities of the various complexes, it is concluded that the stability (half-life) of the whole RTP-Ter complex is not the overriding determinant of arrest, and that the RTP-Ter complex must be actively disrupted, or RTP removed, by the action of the approaching replication fork.


Effects of converting-enzyme inhibition on barrier function in diabetic glomerulopathy.
Differential solute clearances were used to examine the effects of a 90-day course of enalapril on glomerular barrier function in 16 proteinuric patients with diabetic glomerulopathy. By day 90, plasma renin and prorenin became elevated, and arterial pressure declined. Transglomerular passage of dextrans of broad size distribution (radii 28-60 A) was lowered significantly. In a subset of 8 patients, withdrawal of enalapril was followed after an additional 30 days by a return of renin levels and arterial pressure to pretreatment levels. The dextran-sieving profile also returned to baseline, becoming uniformly elevated above treated day-90 levels. A theoretical analysis of the serial dextran-sieving profiles indicated that enalapril shifted glomerular pore size distribution to smaller size. These changes in barrier size selectivity were associated with a reduction in fractional albumin and IgG clearances during enalapril therapy and a subsequent rise in these quantities after its withdrawal; urinary protein excretion rate tended to vary in parallel. We conclude that inhibition of converting enzyme in humans with established diabetic glomerulopathy diminishes glomerular permeability to proteins by enhancing barrier size selectivity. Because neither enalapril therapy nor its withdrawal influenced the glomerular filtration or renal plasma flow rates significantly, we propose that the primary action of enalapril may be to modulate the intrinsic membrane properties of the glomerular barrier.


Quantification of blood volume by electrical impedance tomography using a tissue-equivalent phantom.
An in vivo electrical impedance tomography (EIT) system was designed to accurately estimate quantities of intra-peritoneal blood in the abdominal cavity. For this it is essential that the response is relatively independent of the position of the high conductivity anomaly (blood) in the body. The sensitivity of the system to the introduction of blood-equivalent resistivity anomalies was assessed by using a cylindrical tissue-equivalent phantom. It was found that a satisfactorily uniform response of the system in both radial (transverse) and axial (longitudinal) directions in the phantom could be achieved by filtering resistivity profile images obtained by EIT measurement, and by using extended electrodes to collect data. Post-processing of single impedance images gave rise to a quantity denoted the resistivity index. A filter was then used to remove the remaining radial variation of the resistivity index. It was calculated by evaluating the resistivity index of a number of theoretically calculated images, and constructing a correction filter similar to those used to remove lens imperfections, such as coma, in optical components. The 30% increase in the resistivity index observed when an anomaly was moved to the maximum extent allowed by the filter calculation (0.75 of the phantom radius) was reduced by the filter to 6%. A study of the axial dependence observed in the resistivity index using electrodes extended in the axial direction by +/-5 cm found that the variation in resistivity index with axial position was about half of that observed using small circular electrodes similar to those used in the Sheffield mark I system.


Analysis of heteroplasmy in hypervariable region II of mitochondrial DNA in maternally related individuals.
Mitochondrial DNA sequences have been widely employed for identity investigation. However, the presence of a heteroplasmic site may complicate sequence analysis for forensic purposes when two samples are compared. To study this potential problem, we analyzed the hypervariable region of the displacement loop in five maternally related individuals, that is, grandmother, mother, one son, and two daughters. The results showed that three of them had a heteroplasmic site at nucleotide position (np) 204, located in the hypervariable region II. By using Bayesian inference to assess the significance of the mother-offspring pairs, a likelihood ratio of 1.78 x 10(5) was obtained. Therefore, Bayesian inference does not place the prior odds into the context of the two different likelihood ratios derived from the DNA evidence. On the other hand, the chromatogram of the denaturing high-performance liquid chromatography system proved that the single peak in a sequencing chromatogram at np 204 was, in fact, heteroplasmic in nature. This study demonstrated that heteroplasmy is a common occurrence in tissue from normal individuals and should be taken into account in forensic investigation when samples appear to differ at a single nucleotide position by direct sequencing.


The Role of Myeloid-Derived Suppressor Cells in the Treatment of Pancreatic Cancer.
Pancreatic cancer has the highest mortality rate of all major cancers, with a 5-year survival rate of about 10%. Early warning signs and symptoms of pancreatic cancer are vague or nonexistent, and most patients are diagnosed in Stage IV, when surgery is not an option for about 80%-85% of patients. For patients with inoperable pancreatic cancer, current conventional treatment modalities such as chemotherapy and radiotherapy (RT) have suboptimal efficacy. Tumor progression is closely associated with the tumor microenvironment, which includes peripheral blood vessels, bone marrow-derived inflammatory cells, fibroblasts, immune cells, signaling molecules, and extracellular matrix. Tumor cells affect the microenvironment by releasing extracellular signaling molecules, inducing peripheral immune tolerance, and promoting tumor angiogenesis. In turn, the immune cells of the tumor affect the survival and proliferation of cancer cells. Myeloid-derived suppressor cells are key cellular components in the tumor microenvironment and exert immunosuppressive functions by producing cytokines, recognizing other immune cells, and promoting tumor growth and metastasis. Myeloid-derived suppressor cells are the main regulator of the tumor immune response and a key target for tumor treatments. Since the combination of RT and immunotherapy is the main strategy for the treatment of pancreatic cancer, it is very important to understand the immune mechanisms which lead to MDSCs generation and the failure of current therapies in order to develop new target-based therapies. This review summarizes the research advances on the role of Myeloid-derived suppressor cells in the progression of pancreatic cancer and its treatment application in recent years.


A transcriptomic atlas of mammalian olfactory mucosae reveals an evolutionary influence on food odor detection in humans.
The mammalian olfactory system displays species-specific adaptations to different ecological niches. To investigate the evolutionary dynamics of olfactory sensory neuron (OSN) subtypes across mammalian evolution, we applied RNA sequencing of whole olfactory mucosa samples from mouse, rat, dog, marmoset, macaque, and human. We find that OSN subtypes, representative of all known mouse chemosensory receptor gene families, are present in all analyzed species. Further, we show that OSN subtypes expressing canonical olfactory receptors are distributed across a large dynamic range and that homologous subtypes can be either highly abundant across all species or species/order specific. Highly abundant mouse and human OSN subtypes detect odorants with similar sensory profiles and sense ecologically relevant odorants, such as mouse semiochemicals or human key food odorants. Together, our results allow for a better understanding of the evolution of mammalian olfaction in mammals and provide insights into the possible functions of highly abundant OSN subtypes.


Stable isotope labeling strategy for protein-ligand binding analysis in multi-component protein mixtures.
Described here is a stable isotope labeling protocol that can be used with a chemical modification- and mass spectrometry-based protein-ligand binding assay for detecting and quantifying both the direct and indirect binding events that result from protein-ligand binding interactions. The protocol utilizes an H(2) (16)O(2) and H(2) (18)O(2) labeling strategy to evaluate the chemical denaturant dependence of methionine oxidation in proteins both in the presence and absence of a target ligand. The differential denaturant dependence to the oxidation reactions performed in the presence and absence of ligand provides a measure of the protein stability changes that occur as a result of direct interactions of proteins with the target ligand and/or as a result of indirect interactions involving other protein-ligand interactions that are either induced or disrupted by the ligand. The described protocol utilizes the (18)O/(16)O ratio in the oxidized protein samples to quantify the ligand-induced protein stability changes. The ratio is determined using the isotopic distributions observed for the methionine-containing peptides used for protein identification in the LC-MS-based proteomics readout. The strategy is applied to a multi-component protein mixture in this proof-of-principle experiment, which was designed to evaluate the technique's ability to detect and quantify the direct binding interaction between cyclosporin A and cyclophilin A and to detect the indirect binding interaction between cyclosporin A and calcineurin (i.e., the protein-protein interaction between cyclophilin A and calcineurin that is induced by cyclosporin A binding to cyclophilin A).


Cecal infarction in neonatal calves.
Cecal infarction is an uncommon lesion in calves that results in localized peritonitis and, on occasion, perforation with secondary diffuse peritonitis and death. This lesion in calves has not been described previously. We reviewed the postmortem cases of cecal infarction in dairy calves ≤30 d of age that had been submitted over the course of 5 y to the Tulare branch of the California Animal Health and Food Safety Laboratory System. The area of cecal infarction and the associated lesion margins were examined histologically. Ischemic necrosis of the mucosal side of the cecal wall with various degrees of neutrophilic inflammation of subjacent tissues was found consistently, and thrombosis and vascular occlusion within the areas of necrosis and inflammation was found in 21 of 34 cases. Cecal infarction cases were then compared to controls using a retrospective matched case-control study design. Cases ( n = 34) and controls ( n = 86) were compared with respect to bacteremia (as defined by pure culture of a single bacterial agent from lung and/or liver), species of bacteria isolated, and for the presence of K99 Escherichia coli (calves ≤5 d), attaching and effacing E. coli, Cryptosporidium (calves ≥5 d of age), Salmonella isolation from the intestine, rotavirus, Bovine coronavirus, and Bovine viral diarrhea virus. In addition, the presence of rumenitis or abomasitis, and omphalitis were compared between cases and controls. There were no significant differences in ruminal, abomasal, or umbilical cord tissue inflammation, or pathogen test-positive status between cases and controls.


Contrasting historical and recent gene flow among African buffalo herds in the Caprivi Strip of Namibia.
Population genetic structure is often used to infer population connectivity, but genetic structure may largely reflect historical rather than recent processes. We contrasted genetic structure with recent gene-flow estimates among 6 herds of African buffalo (Syncerus caffer) in the Caprivi Strip, Namibia, using 134 individuals genotyped at 10 microsatellite loci. We tested whether historical and recent gene flows were influenced by distance, potential barriers (rivers), or landscape resistance (distance from water). We also tested at what scales individuals were more related than expected by chance. Genetic structure across the Caprivi Strip was weak, indicating that historically, gene flow was strong and was not affected by distance, barriers, or landscape resistance. Our analysis of simulated data suggested that genetic structure would be unlikely to reflect human disturbances in the last 10-20 generations (75-150 years) because of slow predicted rates of genetic drift, but recent gene-flow estimates would be affected. Recent gene-flow estimates were not consistently affected by rivers or distance to water but showed that isolation by distance appears to be developing. Average relatedness estimates among individuals exceeded random expectations only within herds. We conclude that historically, African buffalo moved freely throughout the Caprivi Strip, whereas recent gene flow has been more restricted. Our findings support efforts to maintain the connectivity of buffalo herds across this region and demonstrate the utility of contrasting genetic inferences from different time scales.


Long-Term Outcomes in Patients With Localized Ewing Sarcoma Treated With Interval-Compressed Chemotherapy on Children's Oncology Group Study AEWS0031.
Clinical trials frequently include multiple end points that mature at different times. The initial report, typically based on the primary end point, may be published when key planned coprimary or secondary analyses are not yet available. Clinical trial updates provide an opportunity to disseminate additional results from studies, published in JCO or elsewhere, for which the primary end point has already been reported.Long-term outcomes from Children's Oncology Group study AEWS0031 were assessed to determine whether the survival advantage of interval-compressed chemotherapy (ICC) was maintained over 10 years in patients with localized Ewing sarcoma (ES). AEWS0031 enrolled 568 eligible patients. Patients were randomly assigned to receive vincristine-doxorubicin-cyclophosphamide and ifosfamide-etoposide alternating once every 3 weeks (standard timing chemotherapy [STC]) versus once every 2 weeks (ICC). For this updated report, one patient was excluded because of uncertainty of original diagnosis. The 10-year event-free survival (EFS) was 70% with ICC compared with 61% with STC (P = .03), and 10-year overall survival (OS) was 76% with ICC compared with 69% with STC (P = .04). There was no difference in the 10-year cumulative incidence of second malignant neoplasms (SMNs; PC [see Data Supplement, online only] = .5). A test for interaction demonstrated that ICC provided greater risk reduction for patients with tumor volume ≥200 mL than for patients with tumors <200 mL, but no evidence for a significant interaction in other subgroups defined by age, primary site, and histologic response. With longer-term follow-up, ICC for localized ES is associated with superior EFS and OS without an increased risk for SMN compared with STC. ICC is associated with improved outcomes even in adverse-risk patient groups.


Cumulative advantage and citation performance of repeat authors in scholarly journals.
Cumulative advantage-commonly known as the Matthew Effect-influences academic output and careers. Given the challenge and uncertainty of gauging the quality of academic research, gatekeepers often possess incentives to prefer the work of established academics. Such preferences breach scientific norms of universalism and can stifle innovation. This article analyzes repeat authors within academic journals as a possible exemplar of the Matthew Effect. Using publication data for 347 economics journals from 1980-2017, as well as from three major generalist science journals, we analyze how articles written by repeat authors fare vis-à-vis less-experienced authors. Results show that articles written by repeat authors steadily decline in citation impact with each additional repeat authorship. Despite these declines, repeat authors also tend to garner more citations than debut authors. These contrasting results suggest both benefits and drawbacks associated with repeat authorships. Journals appear to respond to feedback from previous publications, as more-cited authors in a journal are more likely to be selected for repeat authorships. Institutional characteristics of journals also affect the likelihood of repeat authorship, as well as citation outcomes. Repeat authorships-particularly in leading academic journals-reflect innovative incentives and professional reward structures, while also influencing the intellectual content of science.


Phase variation of manganese oxide in the MnO@ZnO nanocomposite with calcination temperature and its effect on structural and biological activities.
Having powerful antibacterial and antioxidant effects, zinc oxide and manganese oxide nanomaterials are of great interest. Here we have synthesized manganese oxide decorated zinc oxide (MZO) nanocomposites by co-precipitation method, calcined at different temperatures (300-750 °C) and studied various properties. Here the crystalline structure of the nanocomposite and phase change of the manganese oxide are observed with calcination temperature. The average crystalline size increases and the dislocation density and microstrain decrease with the increase in calcined temperature for the same structural features. The formation of composites was confirmed by XRD pattern and SEM images. EDAX spectra proved the high purity of the composites. Here, different biological properties change with the calcination temperature for different shapes, sizes and structures of the nanocomposite. Nanomaterial calcined at 750 °C provides the best anti-microbial activity against Escherichia coli, Salmonella typhimurium, Shigella flexneri (gram-negative), Bacillus subtilis and Bacillus megaterium (gram-positive) bacterial strain at 300 µg/mL concentration. The nanomaterial with calcination temperatures of 300 °C and 450 °C provided better antioxidant properties.


Activity-related summation of pain and functional disability in patients with whiplash injuries.
This study investigated the relation between repetition-induced summation of activity-related pain (RISP) and indicators of functional disability in a sample of 62 individuals who had sustained whiplash injuries. Participants completed measures of pain severity, pain catastrophizing, fear of movement and depression prior to lifting a series of 36 weighted canisters. An index of RISP was computed as the increase in pain reported by participants over successive lifts of the weighted canisters. Measures of functional disability included physical lifting tolerance, self-reported disability and perceived work demands. Regression analyses revealed that the index of RISP accounted for significant variance in measures of lifting tolerance and perceived work demands, even when controlling for age, sex and pain severity. The index of RISP was also significantly correlated with pain catastrophizing and pain duration. The discussion addresses the mechanisms by which physiological and psychological factors might contribute to increases in pain during repeated physical activity. Discussion also addresses whether RISP might represent a risk factor for problematic recovery outcomes following whiplash injury.


Subclinical thyroid dysfunction and hip fracture and bone mineral density in older adults: the cardiovascular health study.
Subclinical thyroid dysfunction is common in the elderly, yet its relationship with hip fracture and bone mineral density (BMD) is unclear. We examined the association between endogenous subclinical hyper- and hypothyroidism and hip fracture and BMD in older adults. A total of 4936 US individuals 65 years old or older enrolled in the Cardiovascular Health Study and not taking thyroid preparations were included. Analyses of incident hip fracture were performed by thyroid status, over a median follow-up of 12 years. A cross-sectional analysis of thyroid status and BMD was performed in a subset of 1317 participants who had dual-energy x-ray absorptiometry scans. Models were adjusted for risk factors and stratified by sex. No association was found between subclinical hypothyroidism and incident hip fracture compared with euthyroidism, when assessed at a single time point or persisting at two time points, in either women [hazard ratio (HR) 0.91, 95% confidence interval (CI) 0.69-1.20 for a single and HR 0.79, 95% CI 0.52-1.21 for two time points] or men (HR 1.27, 95% CI 0.82-1.95 for a single and HR 1.09, 95% CI 0.57-2.10 for two time points). Likewise, no association was found between subclinical hyperthyroidism and incident hip fracture in either sex (HR 1.11, 95% CI 0.55-2.25 in women and HR 1.78, 95% CI 0.56-5.66 in men). No association was found between subclinical thyroid dysfunction and BMD at the lumbar spine, total hip, or femoral neck sites. Our data suggest no association between subclinical hypothyroidism or subclinical hyperthyroidism and hip fracture risk or BMD in older men and women. Additional data are needed to improve the precision of estimates for subclinical hyperthyroidism and in men.


[Anomie in the multigenerational family: children and the care of their elderly parents].
When one of the older parents starts to require intensive and lasting care, to a certain extent an anomic situation will arise in the older family, between children and parents as well as between the children themselves. There is some kind of ambivalence between the generations within the kinship system, which can also be shown historically. It is possible that this ambivalence may even be enlarged by recent structural and cultural developments within the kinship system. Based on a family crisis model a lack of clarity within the older family is shown as to the expectations and obligations to each other. The spiral of the failing competence of the family to handle the care question mostly has its roots in the anomic situation surrounding the organisation of the care activities. To support our anomy hypothesis some problems in taking care of the older parents are analyzed; how they are looked at and handled by the children and how they are negotiated by the parents. Open interviews and some cases are the basis for this analysis and some conclusions in handling the anomic situation are presented.


In vivo and In vitro Transdermal Availability of Ibuprofen using Novel Solubility Enhancing Fluid Nanosized Carrier Systems.
The objective of this study was to explore the benefits of transdermal drug delivery systems as an alternative option for patients who are unable to tolerate oral administration of drugs, such as ibuprofen (IB). To achieve this, nonionic surfactants and three cosolvents were employed to develop new microemulsions (MEs) that contained IB as nanocarriers. The aim was to enhance the solubility and bioavailability of the drug after transdermal administration. The MEs were characterised by droplet size, polydispersity index (PDI), and rheological properties. Furthermore, the flux of IB was evaluated by Franz diffusion cells using excised rat skin and in vivo bioavailability using rats. The results showed that the MEs had ideal viscosity and droplet size below 100 nm. Moreover, using the developed MEs, an improvement in the solubility (170 mg/mL) and flux through the rat skin (94.6 ± 8.0 µg/cm2.h) was achieved. In addition, IB demonstrated a maximum plasma level of 0.064 mg/mL after 8 h of transdermal administration in rats using the ME with an increase in the bioavailability of about 1.5 times in comparison to the commercial IB gel. In conclusion, the developed nonionic MEs containing IB can be ideal nanocarriers and promising formulations for the transdermal administration of IB.


Decrease in age-related tau hyperphosphorylation and cognitive improvement following vitamin D supplementation are associated with modulation of brain energy metabolism and redox state.
In the present study we examined whether vitamin D supplementation can reduce age-related tau hyperphosphorylation and cognitive impairment by enhancing brain energy homeostasis and protein phosphatase 2A (PP2A) activity, and modulating the redox state. Male F344 rats aged 20 months (aged) and 6 months (young) were randomly assigned to either vitamin D supplementation or no supplementation (control). Rats were housed in pairs and the supplementation group (n=10 young and n=10 aged) received subcutaneous injections of vitamin D (1, α25-dihydroxyvitamin D3) for 21 days. Control animals (n=10 young and n=10 aged) received equal volume of normal saline and behavioral testing in the water maze started on day 14 after the initiation of vitamin D supplementation. Tau phosphorylation, markers of brain energy metabolism (ADP/ATP ratio and adenosine monophosphate-activated protein kinase) and redox state (levels of reactive oxygen species, activity of superoxide dismutase, and glutathione levels) as well as PP2A activity were measured in hippocampal tissues. Our results extended previous findings that: (1) tau phosphorylation significantly increased during aging; (2) markers of brain energy metabolism and redox state are significantly decreased in aging; and (3) aged rats demonstrated significant learning and memory impairment. More importantly, we found that age-related changes in brain energy metabolism, redox state, and cognitive function were attenuated by vitamin D supplementation. No significant differences were seen in tau hyperphosphorylation, markers of energy metabolism and redox state in the young animal groups. Our data suggest that vitamin D ameliorated the age-related tau hyperphosphorylation and cognitive decline by enhancing brain energy metabolism, redox state, and PP2A activity making it a potentially useful therapeutic option to alleviate the effects of aging.


Parental Chronic Illness, Internalizing Problems in Young Adulthood and the Mediating Role of Adolescent Attachment to Parents: A Prospective Cohort Study.
Background: Parental chronic illness is associated with an elevated risk for developing social-emotional and behavioral problems in children, in particular internalizing symptoms. This study aimed to investigate the associations between parental chronic illness when participants were adolescents and subsequent internalizing symptoms in young adulthood and whether adolescent attachment to parents or peers mediates these associations. Methods: The study used longitudinal survey data from the Youth and Mental Health Study, a cohort study including a representative sample of youth in central Norway assessed in the period from 1999 to 2000 (mean age 14.9 years) and in 2012 (mean age 27.2 years) (N = 1,266). The data consist of youth self-reports at both time points. Parental chronic illness was reported by the adolescents, quality of attachment was measured using the Inventory of Parent and Peer Attachment (IPPA), and internalizing problems were assessed in young adulthood by using the Adult Self-Report (ASR). Data were analyzed using parallel mediation analyses, controlling for adolescent sex, parental socioeconomic status, and divorce. In addition, separate analyses were conducted for adolescent girls and boys. Results: The total longitudinal effect was significant for both maternal and paternal chronic illness on internalizing problems in young adulthood. The direct effect on internalizing problems was only significant for maternal chronic illness. Attachment to fathers partially mediated the relationship between maternal chronic illness in adolescence and internalizing symptoms in young adulthood, whereas attachment to both mothers and fathers fully mediated the relationship between paternal chronic illness in adolescence and internalizing symptoms in young adulthood. A separate analysis for girls and boys indicated that the results were only significant for girls. Parental chronic illness did not play a significant indirect effect via attachment to peers on internalizing problems. Conclusions: Identifying protective factors in the pathways between parental chronic illness and mental distress in children could guide measures that promote the well-being of the child and family. The study demonstrates the importance of targeting the entire family in chronic illness care.


Epidemiological and Mycological Aspects of Onychomycosis in Dakar (Senegal).
Onychomycosis is a fungal nails infection often caused by yeasts, dermatophytes and molds. It is an important public health concern due to its high prevalence, the problem of diagnostics, and the poor response to treatments. The objective of this study was to evaluate the epidemiological and microbiological profile of onychomycosis diagnosed at the Laboratory of Parasitology-Mycology of the National University Hospital of Fann in Dakar, Senegal, from 2012 to 2016. A retrospective and descriptive study was performed from January 2012 to December 2016 in a patient attending the laboratory of Parasitology-Mycology at the Fann teaching hospital. Socio-demographic, clinical and biological data were collected from the bench registers. Samples from the lesions were tested using direct microscopy and cultured on a Sabouraud-Chloramphenicol and Sabouraud-Chloramphenicol-Actidione medium. A descriptive analysis was done using Stata IC 12 software. The significance level of different tests was set at 5% two-side. A total of 469 patients were included in this study. The mean age of the study population was 33.2 ± 15.2 years, and the sex ratio was 0.52. The prevalence of onychomycosis was 48.4% (227/469). The main clinical presentations were disto-lateral subungual onychomycosis (37.9%) and onyxis (46.5%). Identified fungal species were Candida albicans (42.7%), Candida spp (39.5%), Trichophyton soudanense (10.1%), Fusarium spp (5.3%), and Candida tropicalis (2.6%). Candida albicans was more frequent in subjects over 15 years of age (43.6%) and women (45%). However, Trichophyton soudanense was higher in patients under 15 years old (17.4%) as well as in male subjects (18.8%). In conclusion, onychomycosis is a common cause of consultation in health facilities. Candida albicans and Trichophyton Soudanense are the main fungal species causing onychomycosis. A better understanding of the epidemiology of onychomycosis as well as the spectrum of the pathogen could contribute to improve the management of the infection.


S-Adenosylmethionine decarboxylase.
S-Adenosylmethionine decarboxylase is a key enzyme for the synthesis of polyamines in mammals, plants and many other species that use aminopropyltransferases for this pathway. It catalyses the formation of S-adenosyl-1-(methylthio)-3-propylamine (decarboxylated S-adenosylmethionine), which is used as the aminopropyl donor. This is the sole function of decarboxylated S-adenosylmethionine. Its content is therefore kept very low and is regulated by variation in the activity of S-adenosylmethionine decarboxylase according to the need for polyamine synthesis. All S-adenosylmethionine decarboxylases have a covalently bound pyruvate prosthetic group, which is essential for the decarboxylation reaction, and have similar structures, although they differ with respect to activation by cations, primary sequence and subunit composition. The present chapter describes these features, the mechanisms for autocatalytic generation of the pyruvate from a proenzyme precursor and for the decarboxylation reaction, and the available inhibitors of this enzyme, which have uses as anticancer and anti-trypanosomal agents. The intricate mechanisms for regulation of mammalian S-adenosylmethionine decarboxylase activity and content are also described.


Human metapneumovirus as a major cause of human respiratory tract disease.
Human metapneumovirus (hMPV) is a newly identified paramyxovirus that appears to be one of the most significant and common viral infections in humans. The virus, first isolated in 2001, is a clear cause of lower respiratory tract disease in both the very young and the frail elderly. The virus causes acute wheezing in children or, less commonly, croup or pneumonia. Molecular epidemiology studies have shown that field strains exhibit sufficient sequence diversity to designate 2 subgroups of circulating viruses. Small animal and nonhuman primate models of infection have been described, which will allow studies of pathogenesis and immunity. Recombinant viruses have already been generated by several groups using reverse genetics, which facilitates the study of the biology of the virus and the generation of live attenuated vaccine candidates. Ongoing research promises to elucidate the molecular basis for pathogenesis and immunity of human metapneumovirus infections and to pave the way for rapid vaccine development.


The linker histone homolog Hho1p from Saccharomyces cerevisiae represents a winged helix-turn-helix fold as determined by NMR spectroscopy.
Hho1p is assumed to serve as a linker histone in Saccharomyces cerevisiae and, notably, it possesses two putative globular domains, designated HD1 (residues 41-118) and HD2 (residues 171-252), that are homologous to histone H5 from chicken erythrocytes. We have determined the three-dimensional structure of globular domain HD1 with high precision by heteronuclear magnetic resonance spectroscopy. The structure had a winged helix-turn-helix motif composed of an alphabetaalphaalphabetabeta fold and closely resembled the structure of the globular domain of histone H5. Interestingly, the second globular domain, HD2, in Hho1p was unstructured under physiological conditions. Gel mobility assay demonstrated that Hho1p preferentially binds to supercoiled DNA over linearized DNA. Furthermore, NMR analysis of the complex of a deletion mutant protein (residues 1-118) of Hho1p with a linear DNA duplex revealed that four regions within the globular domain HD1 are involved in the DNA binding. The above results suggested that Hho1p possesses properties similar to those of linker histones in higher eukaryotes in terms of the structure and binding preference towards supercoiled DNA.


Proliferation, bcl-2 expression and angiogenesis in pituitary adenomas: relationship to tumour behaviour.
The prediction of pituitary tumour behaviour, in terms of response to treatment from which can be derived optimal management strategies, is a challenge that has been approached using several different means. Angiogenesis in other tumour types has been shown to be correlated with poor response to treatment and tumour recurrence. The aim of this paper is to assess the role of measurements of cell proliferation and angiogenesis in predicting pituitary tumour behaviour. The proliferative capacity of the tumour was assessed using the Ki-67 labelling index (LI) while bcl-2 expression was used to assess anti-apoptotic pathways. The microvessel density (MVD) was assessed using antibodies to CD31 and factor VIII-related antigen, and with biotinylated ulex europaeus agglutinin I. There was no difference between Ki-67 LI and MVD of functionless tumours that recurred and those that did not, but bcl-2 expression was significantly lower in tumours that subsequently regrew. Macroprolactinomas had significantly higher LI than microprolactinomas and than all other tumours. Cell proliferation and angiogenesis were not related, showing that both processes are under different control mechanisms in pituitary tumours. In contrast there was a positive relationship between markers of angiogenesis and bcl-2 expression in prolactinomas, GH-secreting tumours and non-recurrent functionless tumours with higher levels of bcl-2 expression being found in the more vascular tumours. These findings may suggest that angiogenesis is related to the ability of tumour cells to survive rather than their proliferative activity.


The histologic diagnosis of melanoma.
The pathologic diagnosis of melanoma is of crucial importance because it provides the patient and his or her attending physician with the knowledge of a potentially life-threatening or an almost curable condition. The histopathologic diagnosis of melanoma is based on the simultaneous or systematic assessment of several well-known histopathologic features. This article discusses the author's approach to the histopathologic diagnosis of melanoma for specific situations.


Mechanism of enhanced eicosanoid production by isolated glomeruli from rats with bilateral ureteral obstruction.
Isolated glomeruli from rats with bilateral ureteral obstruction (BUO) of 24-h duration produced significantly greater amounts of prostaglandin (PG) E2 and 6-keto-PGF1 alpha in vitro than glomeruli from sham-operated control (SOC) rats. This increase was abolished by the angiotensin-converting enzyme (ACE) inhibitor, enalaprilat, given in vivo. To elucidate the mechanisms responsible for enhanced eicosanoid production by glomeruli from rats with BUO, we measured the activities of phospholipase (PL) A2 and C and cyclooxygenase in glomeruli isolated from SOC and BUO rats. L-alpha-Phosphatidylcholine (PC)-specific and L-alpha-phosphatidylethanolamine (PE)-specific PLA2 activities were significantly greater in glomerular membranes from rats with BUO than from SOC rats. Likewise, both the activity and amount of cyclooxygenase were significantly greater in glomerular membranes of rats with BUO. Cyclooxygenase and the PE-specific PLA2 in glomerular membranes of rats with BUO remained at the levels seen in SOC rats when animals were treated in vivo before BUO with the ACE inhibitor, enalaprilat, and the thromboxane synthase inhibitor, OKY-046. Thus inhibition of vasoconstrictor formation leads to subsequent inhibition of vasodilator formation. In contrast to PE-specific PLA2, PC-specific PLA2 activities were further increased in glomerular membranes from both SOC and BUO rats pretreated with the two drugs.s The activity of phosphatidylinositol 4,5-bisphosphate-specific phospholipase C (PIP2 PLC) was significantly decreased in glomeruli from rats with BUO compared with SOC rats. We conclude that the increased synthesis of vasodilatory eicosanoids by glomeruli from rats with BUO may be mediated by enhanced activities of PE-specific PLA2 and cyclooxygenase, which are apparently stimulated by the vasoconstrictors angiotensin and thromboxane.


The roles of molecular chaperones in vivo.
Table 1 summarizes the families of chaperones mentioned in this review, and lists their proposed functions. Many of these proteins are named in the accompanying review of Burston and Clarke. Molecular chaperones are proteins which interact with other proteins and help them to reach their final, active conformation. They appear to do this by binding them in an unfolded or partially folded state and subsequently releasing them in an altered form. This property may endow them with several essential or important roles in addition to helping newly synthesized proteins to fold correctly, such as repairing damaged proteins and assisting proteins in membrane translocation. To confirm that a given protein has molecular chaperone activity in vivo, it is necessary to show that interactions between the chaperone and other proteins do occur in the cell, and that loss of the molecular chaperone leads to the accumulation of inactive or precursor protein. The hsp70 protein family are highly conserved and ubiquitous. Genetic studies confirm that their depletion leads to the accumulation of inactive precursor or other proteins, and immunochemical studies show they associate with nascent polypeptides. They are implicated not only in protein folding, but also in protein transport across membranes and reactivation of heat-damaged proteins. The hsp60 proteins are also ubiquitous and very similar in sequence. Those found in bacteria and organelles, such as mitochondria (the GroEL family), are essential at all temperatures, and particularly after heat shock. Their loss or depletion leads to the formation of protein aggregates and eventual cell death. A co-chaperone protein (GroES) is required for their function. Cytosolic homologues (the TCP1 family) are also essential, though not heat-shock induced; they are believed to have a chaperone role in tubulin assembly and their actual role in the cell may be much broader. Many other proteins may have a chaperone function in vivo. Such a function may be specific to a particular substrate (such as the PapD protein in E. coli); others may be more general (such as hsp90 and SecB). Evidence is still needed to demonstrate whether all those proteins which show chaperone behaviour in vitro actually have such a role in vivo. It seems likely that different classes of chaperone may overlap in their specificity, and it is certain that the various proteins classed as molecular chaperones fulfil a wide variety of roles in the cell.


Thyroxine 5'-deiodination mediates norepinephrine-induced lipogenesis in dispersed brown adipocytes.
In euthyroid rats, maximal sympathetic nervous system stimulation (e.g. during cold exposure) results in a 3- to 4-fold increase in brown adipose tissue lipogenesis, a response that is blunted in hypothyroid rats. To further investigate this phenomenon, the role of local type II 5'-deiodinase (5'-DII) was studied in freshly isolated brown adipocytes. In a typical experiment, 1.5 x 10(6) cells were incubated for up to 48 h in a water-saturated 5% CO2-95% O2 atmosphere. After incubation with medium alone or with different concentrations of T4, T3, and/or norepinephrine (NE), lipogenesis was studied by measuring 1) the rate of fatty acid synthesis as reflected by 3H2O incorporation into lipids and 2) the activity of key rate-limiting enzymes, i.e. acetyl coenzyme A carboxylase and malic enzyme, and the results are reported in terms of DNA content per tube. Lipogenesis decreased progressively over time (approximately 40%) when no additions were made to the incubation medium. T4 or T3 partially prevented that inhibition at physiological concentrations (65 x 10[-9] and 0.77 x 10[-9] M, respectively), whereas a receptor-saturating concentration of T3, (154 x 10[-9] M) doubled the lipogenesis rate. The addition of 10(-6) M NE inhibited lipogenesis acutely (approximately 50% by 12 h) and was followed by a progressive stimulation that reached approximately 2-fold by 48 h, but only in the presence of T4. Furthermore, NE did not attenuate T3 (154 x 10[-9] M)-induced lipogenesis. Both the inhibition and the stimulation of lipogenesis caused by NE showed a strong dose-response relationship within the range of 10(-11)-10(-5) M. The role of local 5'-DII was further tested by incubating brown adipocytes with 10(-6) M NE and T4 (65 x 10[-9] M) in the presence of 100 microM iopanoic acid, a potent inhibitor of 5'-DII. Although iopanoic acid did not affect the T3 stimulation of lipogenesis, it did block the approximately 2-fold stimulation of lipogenesis triggered by NE in the presence of T4, confirming the mediation of 5'-DII in this process. In conclusion, lipogenesis in brown adipose tissue is under complex hormonal control, with key roles played by NE, thyroid hormones, and local 5'-DII. As in other tissues, NE-generated signals acutely (12 h) inhibited lipogenesis. However, the presence of the 5'-DII generated enough T3 to stimulate lipogenesis and gradually reverse the short-lived NE-induced inhibition, leading to the 2- to 3-fold response observed at later time points.


A Multi-Task Framework for Monitoring Health Conditions via Attention-based Recurrent Neural Networks.
Monitoring the future health status of patients from the historical Electronic Health Record (EHR) is a core research topic in predictive healthcare. The most important challenges are to model the temporality of sequential EHR data and to interpret the prediction results. In order to reduce the future risk of diseases, we propose a multi-task framework that can monitor the multiple status ofdiagnoses. Patients' historical records are directly fed into a Recurrent Neural Network (RNN) which memorizes all the past visit information, and then a task-specific layer is trained to predict multiple diagnoses. Moreover, three attention mechanisms for RNNs are introduced to measure the relationships between past visits and current status. Experimental results show that the proposed attention-based RNNs can significantly improve the prediction accuracy compared to widely used approaches. With the attention mechanisms, the proposed framework is able to identify the visit information which is important to the final prediction.


Efficacy of repeated administration of intravenous acetaminophen for pain management after total knee arthroplasty.
Intravenous acetaminophen is an integral component of multimodal postoperative pain management. This prospective study aims to assess the efficacy of the repeated administration of intravenous acetaminophen and the impact on postoperative patient satisfaction with postoperative pain management after total knee arthroplasty (TKA). We enrolled 98 patients scheduled for unilateral TKA. Patients were randomly assigned to receive either 1000 mg of intravenous acetaminophen at 6-hour intervals (AAP group) or not to receive intravenous acetaminophen (control group). All patients underwent single-shot femoral nerve block after general anesthesia, as well as intraoperative periarticular infiltration of analgesia prior to implantation. The primary outcome was the postoperative numerical rating scale (NRS) pain score at rest. The NRS score was measured just before the administration of study drugs, immediately after arrival in the ward (time 0), and at 6, 12, 18, 24, and 48 h (time 1 to time 5, respectively) postoperatively. We also evaluated the mean doses of rescue opioid use for 24 h postoperatively. At time 5, the AAP group had significantly improved mean NRS score than controls (3.0 vs. 4.0; P < 0.01). Rescue opioid use was significantly lower in the AAP group for 24 hours compared to controls (0.3 μg vs. 0.9 μg; P < 0.01). Repeated intravenous acetaminophen administration after TKA may provide better analgesia and reduce opioid use.


Two essential splice lariat branchpoint sequences in one intron in a xeroderma pigmentosum DNA repair gene: mutations result in reduced XPC mRNA levels that correlate with cancer risk.
The lariat branch point sequence (BPS) is crucial for splicing of human nuclear pre-mRNA yet BPS mutations have infrequently been reported to cause human disease. Using an inverse RT-PCR technique we mapped two BPS to the adenosine residues at positions -4 and -24 in intron 3 of the human XPC DNA repair gene. We identified homozygous mutations in each of these BPS in two newly diagnosed Turkish families with the autosomal recessive disorder xeroderma pigmentosum (XP). Cells from two severely affected children in family A harbor a homozygous point mutation in XPC intron 3 (-9 T to A), located within the downstream BPS. Using a real-time quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) assay, these cells expressed no detectable (<0.1%) normal XPC message. Instead they expressed an XPC mRNA isoform with deletion of exon 4 that has no DNA repair activity in a host cell reactivation (HCR) assay. In contrast, in cells from three mildly affected siblings in family B, the BPS adenosine located at the -24 position in XPC intron 3 is mutated to a G. Real-time QRT-PCR revealed 3-5% of normal XPC message. These cells from family B had a higher level of HCR than cells from the severely affected siblings in family A, who had multiple skin cancers. Mutations identified in two BPS of the XPC intron 3 resulted in alternative splicing that impaired DNA repair function, thus implicating both of these BPS as essential for normal pre-mRNA splicing. However, a small amount of normal XPC mRNA can provide partial protection against skin cancers.


Echocardiographic assessment of ductal significance: retrospective comparison of two methods.
Patent ductus arteriosus (PDA) in preterm infants is often assessed with echocardiographic parameters, especially colour Doppler ductal diameter and pulsed Doppler flow pattern. Clinical algorithms have been proposed in which PDA treatment is indicated by either large diameter or a particular flow pattern, however it is unknown whether ductal diameter and flow pattern provide equivalent stratification of infants. Retrospectively assess both parameters in 197 echocardiograms from 104 infants (gestational age <31 weeks). Echocardiograms were independently reviewed and the internal colour Doppler diameter of the PDA and the pulsed Doppler flow pattern were characterised for each study (169 records had both parameters recorded). Diameter varied widely within each group but was significantly associated with flow pattern: mean diameter was greatest in the pulmonary hypertension (PH) group (2.6 mm), progressively narrowed across growing and pulsatile groups, and was smallest in the closing group (1.3 mm). When echocardiograms were categorised using previously published diameters, 82.4% of the PH group had diameters >2.0 mm, large diameters predominated in the growing and pulsatile groups but to a progressively smaller extent, and 98.1% of closing group had diameters <2.0 mm. Ductal diameter and flow patterns are significantly associated, consistent with a narrowing of the ductus until closure. Overall, the two parameters are in good agreement but will result in different treatment decisions in some cases. Clinicians might consider using both methods as a cross check against each other, to assist in the management of preterm infants with a clinically detectable PDA.