[Long-term results of retropubic radical prostatectomy].
Retropubic radical prostatectomy (RPPE) is the most aggressive method of treatment in patients with prostate cancer (PC), but maximally radical. The observations of men after RPPE have shown that 10.13% (40 patients) died: half of them (5.06%)--due to progress of PC, the rest because of coexisting diseases. The total, corrected and relapse-free 5 and 10 years survival was 90%, 95%, 65% and 84%, 91%, 55%. The quality of life of the men after RPPE was investigated at long-term periods of followup. RPPE is an effective method of treatment of PC patients. The most frequent complication of this intervention was erectile dysfunction and urinary incontinence which made the quality of life of such patients considerably worse. Postoperative dynamic follow-up of such patients is necessary for early detection of relapses and determination of indications for medicamentous and/or surgical treatments of the complications.


Bone tissue engineering: recent advances and challenges.
The worldwide incidence of bone disorders and conditions has trended steeply upward and is expected to double by 2020, especially in populations where aging is coupled with increased obesity and poor physical activity. Engineered bone tissue has been viewed as a potential alternative to the conventional use of bone grafts, due to their limitless supply and no disease transmission. However, bone tissue engineering practices have not proceeded to clinical practice due to several limitations or challenges. Bone tissue engineering aims to induce new functional bone regeneration via the synergistic combination of biomaterials, cells, and factor therapy. In this review, we discuss the fundamentals of bone tissue engineering, highlighting the current state of this field. Further, we review the recent advances of biomaterial and cell-based research, as well as approaches used to enhance bone regeneration. Specifically, we discuss widely investigated biomaterial scaffolds, micro- and nano-structural properties of these scaffolds, and the incorporation of biomimetic properties and/or growth factors. In addition, we examine various cellular approaches, including the use of mesenchymal stem cells (MSCs), embryonic stem cells (ESCs), adult stem cells, induced pluripotent stem cells (iPSCs), and platelet-rich plasma (PRP), and their clinical application strengths and limitations. We conclude by overviewing the challenges that face the bone tissue engineering field, such as the lack of sufficient vascularization at the defect site, and the research aimed at functional bone tissue engineering. These challenges will drive future research in the field.


Epidemiologic Profile of Erectile Dysfunction in Patients with Systemic Lupus Erythematosus: The Latin American Landscape.
The aim of this study was to describe the prevalence of erectile dysfunction (ED), as well as associated demographic and clinical features, in men with systemic lupus erythematosus (SLE), by means of a systematic, standardized evaluation. We performed a transversal study in 8 tertiary care centers in Latin America. We included male patients ≥ 16 years who fulfilled ≥ 4 American College of Rheumatology criteria for SLE and had regular sexual activity, and evaluated them with the International Index of Erectile Function-5 questionnaire. Relevant demographic, clinical, and serological characteristics were recorded. We included 2 control groups: the first was made up of healthy men and the second of men with autoimmune diseases other than SLE (non-SLE group). We included 590 subjects (174 SLE, 55 non-SLE, and 361 healthy controls). The prevalence of ED in the SLE group was 69%. Mean age in that group was 36.3 ± 1.03 years. Among SLE patients with and without ED, these factors were significantly different: the presence of persistent lymphopenia (p = 0.006), prednisone dose (9.3 ± 1.2 vs 5.3 ± 1.3 mg, p = 0.026), and the Systemic Lupus International Collaborating Clinics damage score (1.25 ± 0.14 vs 0.8 ± 0.16 points, p = 0.042). Independent risk factors for ED in patients with SLE were persistent lymphopenia (OR 2.79, 95% CI 1.37-5.70, p = 0.001) and corticosteroid use in the previous year (OR 2.15, 95% CI 1.37-3.37, p = 0.001). Regardless of comorbidities, treatment (excluding steroids), and type of disease activity, patients with SLE have a high prevalence of ED, especially considering that most patients are young. Recent corticosteroid use and persistent lymphopenia, which could be related to endothelial dysfunction, are risk factors for this complication in men with SLE.


GeneMates: an R package for detecting horizontal gene co-transfer between bacteria using gene-gene associations controlled for population structure.
Horizontal gene transfer contributes to bacterial evolution through mobilising genes across various taxonomical boundaries. It is frequently mediated by mobile genetic elements (MGEs), which may capture, maintain, and rearrange mobile genes and co-mobilise them between bacteria, causing horizontal gene co-transfer (HGcoT). This physical linkage between mobile genes poses a great threat to public health as it facilitates dissemination and co-selection of clinically important genes amongst bacteria. Although rapid accumulation of bacterial whole-genome sequencing data since the 2000s enables study of HGcoT at the population level, results based on genetic co-occurrence counts and simple association tests are usually confounded by bacterial population structure when sampled bacteria belong to the same species, leading to spurious conclusions. We have developed a network approach to explore WGS data for evidence of intraspecies HGcoT and have implemented it in R package GeneMates ( github.com/wanyuac/GeneMates ). The package takes as input an allelic presence-absence matrix of interested genes and a matrix of core-genome single-nucleotide polymorphisms, performs association tests with linear mixed models controlled for population structure, produces a network of significantly associated alleles, and identifies clusters within the network as plausible co-transferred alleles. GeneMates users may choose to score consistency of allelic physical distances measured in genome assemblies using a novel approach we have developed and overlay scores to the network for further evidence of HGcoT. Validation studies of GeneMates on known acquired antimicrobial resistance genes in Escherichia coli and Salmonella Typhimurium show advantages of our network approach over simple association analysis: (1) distinguishing between allelic co-occurrence driven by HGcoT and that driven by clonal reproduction, (2) evaluating effects of population structure on allelic co-occurrence, and (3) direct links between allele clusters in the network and MGEs when physical distances are incorporated. GeneMates offers an effective approach to detection of intraspecies HGcoT using WGS data.


Identification and functional validation of the novel antimalarial resistance locus PF10_0355 in Plasmodium falciparum.
The Plasmodium falciparum parasite's ability to adapt to environmental pressures, such as the human immune system and antimalarial drugs, makes malaria an enduring burden to public health. Understanding the genetic basis of these adaptations is critical to intervening successfully against malaria. To that end, we created a high-density genotyping array that assays over 17,000 single nucleotide polymorphisms (∼ 1 SNP/kb), and applied it to 57 culture-adapted parasites from three continents. We characterized genome-wide genetic diversity within and between populations and identified numerous loci with signals of natural selection, suggesting their role in recent adaptation. In addition, we performed a genome-wide association study (GWAS), searching for loci correlated with resistance to thirteen antimalarials; we detected both known and novel resistance loci, including a new halofantrine resistance locus, PF10_0355. Through functional testing we demonstrated that PF10_0355 overexpression decreases sensitivity to halofantrine, mefloquine, and lumefantrine, but not to structurally unrelated antimalarials, and that increased gene copy number mediates resistance. Our GWAS and follow-on functional validation demonstrate the potential of genome-wide studies to elucidate functionally important loci in the malaria parasite genome.


Abdominal pregnancy: a case report and literature review.
We report a rare and unusual case of an Abdominal pregnancy that was delivered successfully at term. It is the first documented case in the local literature in almost 30 years. What makes it even more exceptional is that despite the high mortality associated with the condition, both mother and baby survived the ordeal. She subsequently delivered another child, 2 years later by an elective lower segment caesarean section and had a tubal ligation performed concomitantly. A review of the topic and its management is also presented.


A genetic mosaic screen of essential zygotic genes in Caenorhabditis elegans.
We have devised a simple genetic mosaic screen, which circumvents the difficulties posed by phenotypic analysis of early lethal mutants, to analyze essential zygotic genes in Caenorhabditis elegans. The screen attempts to distinguish genes involved in cell type and/or lineage specific processes such as determination, differentiation or morphogenesis from genes involved in general processes such as intermediary metabolism by using the pattern of gene function to classify genes: genes required in one or a subset of early blastomeres may have specific functions, whereas genes required in all early blastomeres may have general functions. We found that 12 of 17 genes examined function in specific early blastomeres, suggesting that many zygotic genes contribute to specific early processes. We discuss the advantages and limitations of this screen, which is applicable to other regions of the C. elegans genome.


Babies and machines that go 'beep': first-year nursing students' preferred areas of future practice.
Students of nursing enter their programmes of study with preconceived ideas of what a career in their chosen profession will entail. The literature suggests that images from the media and past experiences contribute to these perceptions. Although it is positive images of the profession that will usually attract an individual to a career in nursing, often more negative perceptions will direct students away from potentially rewarding areas of specialization. This paper describes career projections of nursing students enrolled in the first year of four preservice nursing programmes at the rural campus of one Australian university. Part of a larger study, the data reported here indicate that most respondents intend to practice in the areas of midwifery, paediatrics and emergency nursing. Oncology, community nursing, aged care and mental health nursing all ranked poorly across three rounds of surveys. These findings have implications for practicing nurses and nurse educators who seek to dispel inaccurate images of these important specializations.


Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches.
Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entanglements generated during DNA replication and repair. While the catalytic cycle of Type 1A topoisomerases has been long-known to involve an enzyme-bridged ssDNA gate that allows strand passage, a deeper mechanistic understanding of these enzymes has only recently begun to emerge. This knowledge has been greatly enhanced through the combination of biochemical studies and increasingly sophisticated single-molecule assays based on magnetic tweezers, optical tweezers, atomic force microscopy and Förster resonance energy transfer. In this review, we discuss how single-molecule assays have advanced our understanding of the gate opening dynamics and strand-passage mechanisms of Type 1A topoisomerases, as well as the interplay of Type 1A topoisomerases with partner proteins, such as RecQ-family helicases. We also highlight how these assays have shed new light on the likely functional roles of Type 1A topoisomerases in vivo and discuss recent developments in single-molecule technologies that could be applied to further enhance our understanding of these essential enzymes.


Psychotropic drug prescription for nursing home residents with dementia: prevalence and associations with non-resident-related factors.
To determine psychotropic drug prescription rates in nursing home residents with dementia and to identify associations with the so far understudied psychosocial non-resident-related factors. A cross-sectional, observational, exploratory design as part of PROPER I (PRescription Optimization of Psychotropic drugs in Elderly nuRsing home patients with dementia). Participants were 559 nursing home residents with dementia, 25 physicians, and 112 nurses in the Netherlands. Psychotropic drug prescription, non-resident-related and known resident-related variables were measured to operationalize the themes of our previous qualitative analysis. Fifty-six percent of residents were prescribed any psychotropic drug, 25% antipsychotics, 29% antidepressants, 15% anxiolytics, and 13% hypnotics, with large differences between the units. Multivariate multilevel regression analyses revealed that antipsychotic prescription was less likely with higher physicians' availability (odds ratio 0.96, 95% confidence interval 0.93-1.00) and that antidepressant prescription was more likely with higher satisfaction of nurses on resident contact (odds ratio 1.50, 95% confidence interval 1.00-2.25). Resident-related factors explained 6%-15% of the variance, resident- and non-resident-related factors together 8%-17%. Prescription rates for antipsychotics are similar compared to other countries, and relatively low for antidepressants, anxiolytics, and hypnotics. Our findings indicate that improvement of prescribing could provisionally best be targeted at resident-related factors.


Lidocaine spray administration during transrectal ultrasound guided prostate biopsy modified the discomfort and pain of the procedure: results of a randomized clinical trial.
We report the results of a study about the possible benefit of lidocaine spray perineal administration before transrectal ultrasound guided biopsy of the prostate. Many patients frequently report some kind of discomfort and (or) pain during this procedure, that when pain is severe, may be necessary to interrupt. Between September 2007 and October 2009 372 consecutive male patients with elevate PSA and (or) abnormal digital rectal and (or) suspect TRUS were scheduled for prostate biopsy and divided in 3 groups. Group 1 (n = 98) underwent intrarectal instillation of a lidocaine/prilocaine cream (EMLA R), Group 2 (n = 126) of a 2.5% lidocaine gel, and Group 3 (n = 148) administration of a lidocaine spray (10 gr/100 ml) before the procedure. A verbal numerical pain score (VNS) from = 0 no discomfort to 10 = severe pain was admnistrated to the biopsied patients who were asked to evaluate separately the degree of pain associated with the insertion of the probe and the manoeuvres associated with it and the degree of pain associated with the biopsy. The mean value of pain VNS in patients of the first group was respectively 5.3 (2-8) for the insertion of the probe (first question) and 3.2 (2-7) for the biopsy by itself (second question), whereas in the second group it was 6.2 (4-9) and 3.8 (3-8), and in the third group 3.1 (1-6) and 2.8 (0-6). Pain score results showed that the use of intrarectal lidocaine spray provided significantly better pain control than cream and anaesthetic gel. Our pain score data suggests that lidocaine spray provides efficient patient comfort during prostate biopsy by reducing pain both during probe insertion and insertion of the needle through the prostate gland. The use of lidocaine spray makes an excellent alternative, causing a reduction of anal sphincter tone with better patient compliance and tolerability to the ultrasound probe during biopsies with an optimization in terms of cost-effectiveness of the procedure.


Effects of the COVID-19 pandemic on the physical activity and screen time habits of children aged 11-13 years in Sweden.
Physical activity (PA), exercise, sedentary behavior and screen time are lifestyle factors that have been shown to significantly impact child health in different ways. These lifestyle factors were affected to different degrees by global restrictions during the COVID-19 pandemic. We investigated PA and screen time in a cohort of Swedish children in both 2019 and 2021, before and during the pandemic. Adolescents born in 2008 in Halland, Sweden, and included in a previous longitudinal birth cohort study were invited to take part in follow-up questionnaires about PA, screen time and COVID-19. A total of 1041 children aged 11 (in 2019) and 13 years (in 2021) replied and 777 of them answered on both occasions. Most children (42.1%) reported that their leisure time PA was unchanged from 2019 to 2021. Compared to unchanged PA 33.9% exercised more often (p = 0.011) and 23.9% exercised less (p < 0.001), both differences statistically significant. Roughly, 43.2% of boys and 34.9% of girls in 2021 exercised so that they became breathless or broke a sweat at least 4 times a week not counting physical education in school, corresponding figures for 2019 were 38.2% for boys and 35.2% for girls. The majority of children were able to continue attending leisure time sports clubs during the pandemic, but participation decreased from 88.3% to 76.3% from 11 to 13 years of age. Most reported that sports club routines changed during the pandemic, but only 40.9% reported fewer practice opportunities. Attending a sports club gave greater protection against loss of PA during the pandemic than not belonging to one (41.0% vs. 23.2%, p < 0.001). The majority (71.1%) of children spent more time on screens in 2021 than 2019, with a mean increase of 9.4 h (95% CI 8.6 to 10.2 h) from 20.7 to 30.1 hours per week (p < 0.001) during the study. Swedish children largely maintained their levels of PA during the pandemic at 13 years of age and these were possibly safeguarded by the comparably mild pandemic restrictions in Sweden in 2021. However, they did increase their screen time between 11 and 13 years of age.


Obesity Related Coronary Microvascular Dysfunction: From Basic to Clinical Practice.
Obesity related coronary microvascular disease is a medical entity which is not yet fully elucidated. The pathophysiological basis of coronary microcirculatory dysfunction consists of a heterogeneous group of disorders with individual morphologic/functional/clinical presentation and prognosis. Coronary microcirculatory changes include mechanisms connected with vascular dysfunction, as well as extravascular and vasostructural changes in responses to neural, mechanical, and metabolic factors. Cardiometabolic changes that include obesity, dyslipidemia, diabetes mellitus type II, and hypertension are associated with atherosclerosis of epicardial coronary arteries and/or microvascular coronary dysfunction, with incompletely understood underlying mechanisms. In obesity, microvascular disease is mediated via adipokines/cytokines causing chronic, subclinical inflammation with (a) reduced NO-mediated dilatation, (b) changed endothelial- and smooth muscle-dependent vasoregulating mechanisms, (c) altered vasomotor control with increased sympathetic activity, and (d) obesity related hypertension with cardiomyocytes hypertrophy and impaired cardiac vascular adaptation to metabolic needs. From a clinical point of view it can present itself in acute or chronic form with different prognosis, as a practice problem for real-life diagnosis and treatment.


A neutralizing monoclonal antibody to bovine rotavirus VP8 neutralizes rotavirus infection without inhibiting virus attachment to MA-104 cells.
VP8*, the N-terminal cleavage product of rotavirus VP4, contains the virus neutralizing epitopes in the hemagglutination domain. To map the neutralizing epitope, we developed monoclonal antibodies specific for VP4 of bovine rotavirus C486 (BRV). A neutralizing escape mutant was generated by one of these monoclonal antibodies (2E8) and a point mutation (Glu-->Asp) was found at aa 116 of VP8*. To investigate the effect of this mutation on the cellular binding and hemagglutination activities, the VP8* genes of the escape mutant and wild type (WT) virus were expressed in E. coli and their functional activities were compared. Both the escape mutant and WT virus VP8* showed hemagglutination and MA-104 cell binding activities. However, hemagglutination activity of the WT virus VP8* was inhibited by 2E8, but that of the escape mutant VP8* was not. These data indicate that the neutralizing epitope is located in the HA domain but is not critical for rotavirus attachment to MA-104 cells. To understand virus neutralization, radiolabelled BRV was incubated with 2E8 and the distribution of radioactivity in a CsCI density gradient was analysed as was the morphology of the virions in peak fractions. Interaction of 2E8 with rotavirus led to virus morphological changes with a concomitant shift in buoyant density. These data suggest that aa 116 influences the binding of 2E8 which in turn may alter virus integrity.


Cyclin-dependent kinase 1 gene expression is associated with poor prognosis in gastric carcinoma.
A low expression level of cyclin-dependent kinase (Cdk) inhibitor p27 is associated with high aggressiveness and poor prognosis of various carcinomas. Human Cdk subunit 1 (Cks1), as well as S-phase kinase-associated protein 2 (Skp2), is an essential and specific factor in the p27 proteolysis by SCF(Skp2) ubiquitin ligase. The purpose of this study is to clarify the clinical significance of Cks1 expression and the relationship between Cks1 and p27 expression in gastric carcinomas. We measured Cks1 expression using quantitative reverse transcription-PCR in 76 human gastric carcinomas and p27 expression using immunohistochemistry in 28 cases. Moreover, we established Cks1- and/or Skp2-transfected gastric carcinoma cell lines and assessed the relationship between Cks1, Skp2, and p27 expression using quantitative reverse transcription-PCR and Western blot analysis. Cks1 high expression was correlated with poor prognosis (P < 0.05) and Cks1 expression was inversely correlated with the expression level of p27 protein in gastric carcinomas (P < 0.05). Using combined Skp2 data [T-a. Masuda, Cancer Res., 62: 3819-3825, 2002], 88.9% of the Cks1/Skp2 double-high cases expressed a low level of p27 protein and showed the poorest prognosis (P < 0.05). Western blot analysis showed that Cks1/Skp2-cotransfected cells expressed a much lower level of p27 protein than the controls. These findings indicate that Cks1, as well as Skp2, regulates the expression level of p27 protein in gastric carcinomas. Cks1 could play an important role in gastric carcinoma progression and would be a novel target for the treatment of gastric carcinomas as well as a strong prognostic marker.


The transition state structure for coupled binding and folding of disordered protein domains.
Intrinsically disordered proteins are abundant in the eukaryotic proteome, and they are implicated in a range of different diseases. However, there is a paucity of experimental data on molecular details of the coupled binding and folding of such proteins. Two interacting and relatively well studied disordered protein domains are the activation domain from the p160 transcriptional co-activator ACTR and the nuclear co-activator binding domain (NCBD) of CREB binding protein. We have analyzed the transition state for their coupled binding and folding by protein engineering and kinetic experiments (Φ-value analysis) and found that it involves weak native interactions between the N-terminal helices of ACTR and NCBD, but is otherwise "disordered-like". Most native hydrophobic interactions in the interface between the two domains form later, after the rate-limiting barrier for association. Linear free energy relationships suggest a cooperative formation of native interactions, reminiscent of the nucleation-condensation mechanism in protein folding.


Recognizing global burden of cardiovascular disease and related chronic diseases.
Cardiovascular disease is the leading cause of death worldwide, affecting not only high-income but also low- and middle-income countries. Nearly 80 percent of all estimated cardiovascular disease-related deaths worldwide now occur in low- and middle-income countries, where nearly 30 percent of all deaths are attributable to cardiovascular disease. The health burden of cardiovascular disease and other chronic diseases is also accompanied by a significant deleterious economic impact at the level of both national economies and households. The global trends in the health and economic burden of cardiovascular disease provide a compelling argument in support of prioritizing urgent yet carefully planned efforts to prevent and control cardiovascular disease worldwide-and especially in low- and middle-income countries. After decades of escalating efforts to draw attention to the high burden of cardiovascular disease and other chronic diseases, this critically important issue is now emerging as a more central part of the global health and development agenda. The breadth of behavioral, biological, social, environmental, and systems-level factors that contribute to cardiovascular disease necessitates multisectoral approaches across the lifecourse that promote healthful lifestyles, reduce risk, and reduce cardiovascular-disease morbidity and mortality through the delivery of quality health care services. Given that the complex interactions among the determinants of cardiovascular disease vary in different contexts, real progress in control efforts will come through approaches that are driven by a country's disease burden and risk profile, capacities, resources, and priorities-approaches that are led by a country's key decision-makers and stakeholders, including governments, civil society, the private sector, and communities. Many countries are already establishing efforts to address chronic diseases. In addition to these locally driven efforts, success will require active engagement and sustained action from a wide array of stakeholders operating at global and regional levels.


Promoter methylation of DAP-kinase: association with advanced stage in non-small cell lung cancer.
Death associated protein (DAP)-kinase is a 16 kDa calmodulin-dependent serine/threonine kinase that carries a death domain at its C-terminus. DAP-kinase functions as a positive mediator of apoptosis that is induced by interferon-gamma. Recent studies suggest that DAP-kinase is involved in tumor metastasis and that it can be inactivated by methylation of CpG islands in the promoter region of the gene in some human tumors. However, little is known about the factors that are associated with the occurrence of DAP-kinase promoter methylation. We investigated both the possible associations of tobacco carcinogen and asbestos exposure with DAP-kinase promoter methylation, and the demographic and clinical factors associated with DAP-kinase promoter methylation in non-small cell lung cancer (NSCLC). One hundred and eighty-five patients diagnosed with NSCLC undergoing surgical resection from June, 1992 through December, 1996 at Massachusetts General Hospital participated in this study. Methylation-Specific PCR (MSP), performed using fresh-frozen tissue, was used to determine the methylation status of the promoter region of the DAP-kinase gene. Forty-seven (25%) of 185 tumors showed DAP-kinase promoter methylation. There was a significant association between methylation and an advanced pathologic stage (P=0.003, Fisher's exact test). Methylation of the DAP-kinase promoter was also associated with an increase in tumor size (P=0.009, Fisher's exact test) and lymph node involvement (P=0.04). No association was found between promoter methylation of DAP-kinase and k-ras or p53 mutation. In addition there was no association with a history of exposure to tobacco or asbestos. Controlling for age, sex, and histology, the odds ratios describing the association of DAP-kinase hypermethylation with stage were 2.70 (1.13--6.45), 3.11 (1.37--7.08) and 7.77 (1.21--50.03) in stages II, III and IV, respectively. Stage I cases with DAP-kinase promoter methylation had worse overall survival, but with the small sample size and limited follow-up this did not reach statistical significance. Our findings suggest that methylation of the promoter region of the DAP-kinase gene is not associated with exposure to tobacco or asbestos. However, they strongly suggest that DAP-kinase may be important in the progression of non-small cell lung cancer from early to late stage disease.


Interaction between ICOS-B7RP1 and B7-CD28 costimulatory pathways in alloimmune responses in vivo.
The B7-CD28 pathway is one of the foremost costimulatory pathways involved in T-cell activation. Recently, a number of additional costimulatory pathways have been described and preliminary data suggest that they play important roles in alloimmunity. However, the interactions between these different pathways are not well understood. We studied the effect of targeting ICOS ligand, B7RP1, in a rat cardiac transplant model, with and without concomitant blockade of the B7 pathway using CTLA4Ig. In a fully mismatched WF to LEW vascularized cardiac allograft model, without therapy, grafts were acutely rejected (MST 10.8 +/- 1.6 days). Early (day of transplant) B7RP1 blockade with ICOSIg alone had little effect on graft survival and rather than being additive with B7 blockade, ICOSIg abrogated the prolonged graft survival induced by CTLA4Ig treatment. By contrast, delayed (day 2 post-transplant) blockade of B7RP1 did not have such an effect. These findings were not related to cytokine deviation but may be in part related to the pattern of down-regulation of B7.2 expression following early B7RP1 blockade. This is the first report describing the complex interactions between ICOS-B7RP1 and B7-CD28 costimulatory pathways in alloimmunity in vivo.


Learning in the Rodent Motor Cortex.
The motor cortex is far from a stable conduit for motor commands and instead undergoes significant changes during learning. An understanding of motor cortex plasticity has been advanced greatly using rodents as experimental animals. Two major focuses of this research have been on the connectivity and activity of the motor cortex. The motor cortex exhibits structural changes in response to learning, and substantial evidence has implicated the local formation and maintenance of new synapses as crucial substrates of motor learning. This synaptic reorganization translates into changes in spiking activity, which appear to result in a modification and refinement of the relationship between motor cortical activity and movement. This review presents the progress that has been made using rodents to establish the motor cortex as an adaptive structure that supports motor learning.


Abnormalities of Mitochondrial Dynamics in Neurodegenerative Diseases.
Neurodegenerative diseases are incurable and devastating neurological disorders characterized by the progressive loss of the structure and function of neurons in the central nervous system or peripheral nervous system. Mitochondria, organelles found in most eukaryotic cells, are essential for neuronal survival and are involved in a number of neuronal functions. Mitochondrial dysfunction has long been demonstrated as a common prominent early pathological feature of a variety of common neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and Huntington's disease (HD). Mitochondria are highly dynamic organelles that undergo continuous fusion, fission, and transport, the processes of which not only control mitochondrial morphology and number but also regulate mitochondrial function and location. The importance of mitochondrial dynamics in the pathogenesis of neurodegenerative diseases has been increasingly unraveled after the identification of several key fusion and fission regulators such as Drp1, OPA1, and mitofusins. In this review, after a brief discussion of molecular mechanisms regulating mitochondrial fusion, fission, distribution, and trafficking, as well as the important role of mitochondrial dynamics for neuronal function, we review previous and the most recent studies about mitochondrial dynamic abnormalities observed in various major neurodegenerative diseases and discuss the possibility of targeting mitochondrial dynamics as a likely novel therapeutic strategy for neurodegenerative diseases.


Identification and characterization of a phosphodiesterase that inversely regulates motility and biofilm formation in Vibrio cholerae.
Vibrio cholerae switches between free-living motile and surface-attached sessile lifestyles. Cyclic diguanylate (c-di-GMP) is a signaling molecule controlling such lifestyle changes. C-di-GMP is synthesized by diguanylate cyclases (DGCs) that contain a GGDEF domain and is degraded by phosphodiesterases (PDEs) that contain an EAL or HD-GYP domain. We constructed in-frame deletions of all V. cholerae genes encoding proteins with GGDEF and/or EAL domains and screened mutants for altered motility phenotypes. Of 52 mutants tested, four mutants exhibited an increase in motility, while three mutants exhibited a decrease in motility. We further characterized one mutant lacking VC0137 (cdgJ), which encodes an EAL domain protein. Cellular c-di-GMP quantifications and in vitro enzymatic activity assays revealed that CdgJ functions as a PDE. The cdgJ mutant had reduced motility and exhibited a small decrease in flaA expression; however, it was able to produce a flagellum. This mutant had enhanced biofilm formation and vps gene expression compared to that of the wild type, indicating that CdgJ inversely regulates motility and biofilm formation. Genetic interaction analysis revealed that at least four DGCs, together with CdgJ, control motility in V. cholerae.


Role of the mitochondrial genome during early development in mice. Effects of ethidium bromide and chloramphenicol.
The role of the mitochondrial genome in early development and differentiation was studied in mouse embryos cultured in vitro from the two to four cell stage to the blastocyst (about 100 cells). During this period the mitochondria undergo morphological differentiation: progressive enlargement followed by an increase in matrix density, in number of cristae, and in number of mitochondrial ribosomes. Mitochondrial ribosomal and transfer RNA synthesis occurs from the 8 to 16 cell stage on and contributes to the establishment of a mitochondrial protein-synthesizing system. Inhibition of mitochondrial RNA- and protein-synthesis by 0.1 microg/ml of ethidium bromide or 31.2 microg/ml of chloramphenicol permits essentially normal embryo development and cellular differentiation. Mitochondrial morphogenesis is also nearly normal except for the appearance of dilated and vesicular cristae in blastocyst mitochondria. Such blastocysts are capable of normal postimplantation development when transplanted into the uteri of foster mothers. Higher concentrations of these inhibitors have general toxic effects and arrest embryo development. It is concluded that mitochondrial differentiation in the early mouse embryo occurs through the progressive transformation of the preexisting mitochondria and is largely controlled by the nucleocytoplasmic system. Mitochondrial protein synthesis is required for the normal structural organization of the cristae in blastocyst mitochondria. Embryo development and cellular differentiation up to the blastocyst stage are not dependent on mitochondrial genetic activity.


[Interspecific recombinants of Bacillus thuringiensis x Bacillus cereus].
The possibility of interspecies recombination was shown by using protoplast fusion method. The Bacillus thuringiensis var. galleriae strain 48S Thi Nic Gua Rifr Strr and 56R Gua Rifr, and also Bac. cereus carrying the plasmid pBC16 responsible for resistance to tetracycline (150 mcg/ml) were used. Recombinants were selected on the medium containing rifampicin and tetracycline. They were shown to combine the properties of both parents. The majority of recombinants were resistant to phages Tg4 and Td15 and represented the mean level of sensitivity to phages Tg12, Tg13 and Td14. Examination of the plasmid profiles of recombinants revealed that their resistance to tetracycline was due to the plasmid with mobility analogous to pBC16. It was concluded that the protoplast fusion method can be used to obtain recombinants between relatively remote species of microorganisms.


Impact of a Parent Video Viewing Program in the Neonatal Intensive Care Unit.
Purpose: Video visits, or televisits, have become increasingly popular across various medical subspecialties. Within the University of California, Davis, Neonatal Intensive Care Unit, a video visitation program known as FamilyLink allows families to remotely view their babies when they are otherwise unable to visit. This study aimed to explore parents' perceived effects of video camera use as well as the relationship of video visit use with rates of breast milk feedings at hospital discharge. Materials and Methods: Families enrolled in this study completed a series of two identical surveys that gathered self-reported data on their experiences during their infant's hospitalization. Comparisons were made considering whether the FamilyLink program was utilized during the admission as well as changes in self-reported experiences over the time course of the hospital admission. The type of enteral feeding at discharge was recorded and reviewed for each baby. Results: Of 100 families enrolled in the study, 30 were found to have used FamilyLink to visit with their baby. The use of FamilyLink was associated with survey findings of sustained intention to breastfeed or provide breast milk to the baby, as well as increased perceived parental involvement in the baby's care. Improved rates of breast milk feedings at the time of discharge were also found among babies whose families conducted televisits using FamilyLink. Conclusions: Video viewing in the NICU has effected a positive impact on breast milk feedings and parents' feelings of involvement during the admission, with the potential to further improve on families' experiences with a hospitalized baby.


The Impact of Olfactory Dysfunction on the Psychological Status and Quality of Life of Patients with Nasal Obstruction and Septal Deviation.
Introduction Olfactory dysfunction may be present in patients with nasal obstruction and septal deviation. The impact of olfactory dysfunction on the psychological profile and quality of life (QoL) of these patients remains unexplored. Objective The present study aimed to investigate the emotional status and QoL of patients with olfactory dysfunction and septal deviation and to identify predictors associated with clinically significant improvement of psychological status and QoL, focusing mainly on the role of olfactory recovery after septoplasty. Methods The olfactory function was quantitatively assessed using the ''Sniffin' sticks'' test (Burghart Messtechnik GmbH, Wedel, Germany) in 60 patients and 25 controls enrolled in this prospective study. The participants completed validated questionnaires specific for general health (Short Form-36), nasal-symptom related QoL (SinoNasal Outcome Test-22), olfaction-associated QoL (Questionnaire of Olfactory Deficits) and for assessing their psychological state (Short Anxiety Screening Test and Beck Depression Inventory) preoperatively and 6 months postoperatively. The patients used the Glasgow Benefit Inventory to evaluate their personal benefit after septoplasty with. Results Septoplasty led to significantly improved olfactory function. Patients with olfactory impairment had significantly lower nasal-symptom related QoL, higher stress levels, and more depressive mood compared with normosmics and controls before and after septoplasty. Postoperatively, personal benefit from surgery was higher in normosmic patients. Improvement of nasal-symptom related QoL was significantly associated with higher likelihood of clinically significant improvement of patients' psychological profile and more personal benefit from surgery. Olfactory dysfunction was negatively correlated with the emotional status of the patients. Conclusion Olfactory dysfunction appears to significantly affect the psychological status of patients with nasal obstruction, and olfactory recovery improves the patients' perception of personal benefit from septoplasty.


Radon in harvested rainwater at the household level, Palestine.
The main objective of this study was to assess Radon concentration in the harvested rainwater (HRW) at the household level in Yatta area, Palestine. HRW is mainly used for drinking as it is the major source of water for domestic uses due to water scarcity. Ninety HRW samples from the household cisterns were collected from six localities (a town and five villages) and Radon concentrations were measured. The samples were randomly collected from different households to represent the Yatta area. Fifteen samples were collected from each locality at the same day. RAD7 device was used for analysis and each sample was measured in duplicate. Radon concentrations ranged from 0.037 to 0.26 Bq/L with a mean ± standard deviation of 0.14 ± 0.06 Bq/L. The estimated annual effective radiation doses for babies, children and adults were all far below the maximum limit of 5 mSvy-1 set by the National Council on Radiation Protection and Measurements.


The mouse Plk gene: structural characterization, chromosomal localization and identification of a processed Plk pseudogene.
The Plk gene encodes a serine/threonine protein kinase believed to be important for the normal progression of mammalian cells through the cell cycle. In this paper, we report the genomic organization of the mouse Plk gene. The mouse Plk gene encompasses 16 kb of the mouse genome and is organised into 10 exons. Based on homology with the human PLK1 promoter region, the putative mouse promoter region includes a CCAAT motif but lacks the conventional TATA motif. The proposed promoter region contains consensus binding sites for several transcriptional regulators, including Sp1 and AP2. In addition to the active copy of Plk, Plk exists as a processed pseudogene. Using RFLP analysis, we have localized the active Plk gene to mouse Chromosome 7 and the processed pseudogene to mouse Chromosome 5. Southern blot analysis of DNA from a limited number of other mammalian species suggests that the duplication is confined to the mouse. Parsimony analysis suggests that the gene duplication leading to the mouse Plk pseudogene occurred after the rat-mouse split.


Inhibition of nerve growth factor/tyrosine kinase receptor A signaling ameliorates airway remodeling in chronic allergic airway inflammation.
The molecular mechanism leading to airway remodeling in patients with allergic asthma is not fully understood. We determined the role of nerve growth factor/tyrosine kinase receptor A signaling in airway remodeling in chronic allergic airway inflammation, and proved that inhibited nerve growth factor (NGF) production ameliorates airway remodeling during chronic allergic airway inflammation. Six- to eight-week-old female BALB/c mice were used in this study. Mice were randomized into four groups: phosphate buffer saline (PBS) control group (n = 10); chronic asthmatic group (n = 12); anti-NGF group (n=12); and anti-TrkA group (n=12). First, to determine the impact of NGF on airway remodeling, antibody-blocking experiments were performed in a chronic allergic murine model characterized by matrix deposition in the subepithelial. Secondly, the number of eosinophils, macrophages, neutrophils and the total number of cells in bronchoalveolar lavage fluid (BALF) was counted. Thirdly, growth-associated protein 43 (GAP43) and NGF protein expression was measured by western blot. It was shown that the number of eosinophils and the total inflammatory cells, NGF and GAP43 protein expression in BALF were markedly higher in asthma group, compared to the other groups. And given anti-NGF or anti-TrkA antibody treatment can reduced GAP43 expression and collagen deposition in the airway. NGF triggers wound healing process and airway remodeling by inducing GAP43 production dependent on TrkA in a mouse model of chronic experimental asthma. Controlling epithelial NGF production might be an efficient therapeutic target to prevent allergic asthma.


Venous thromboembolism risk in psychiatric in-patients: a multicentre cross-sectional study.
Aims and methodWe assessed venous thromboembolism (VTE) risk, barriers to prescribing VTE prophylaxis and completion of VTE risk assessment in psychiatric in-patients. This was a cross-sectional study conducted across three centres. We used the UK Department of Health VTE risk assessment tool which had been adapted for psychiatric patients. RESULTS: Of the 470 patients assessed, 144 (30.6%) were at increased risk of VTE. Patients on old age wards were more likely to be at increased risk than those on general adult wards (odds ratio = 2.26, 95% CI 1.51-3.37). Of those at higher risk of VTE, auditors recorded concerns about prescribing prophylaxis in 70 patients (14.9%). Only 20 (4.3%) patients had a completed risk assessment.Clinical implicationsMental health in-patients are likely to be at increased risk of VTE. VTE risk assessment is not currently embedded in psychiatric in-patient care. There is a need for guidance specific to this population.Declaration of interestNone.


The ethics of 'fail first': guidelines and practical scenarios for step therapy coverage policies.
In an effort to control health costs, payers are increasingly turning to step therapy (or "fail first") policies in pharmacy benefit design. These policies restrict coverage of expensive therapies unless patients have already failed treatment with a lower-cost alternative. More than other utilization management tools such as formulary tiering, step therapy raises important ethical concerns regarding the proper balance between cost control and the ability of patients and clinicians to tailor care to the needs of the individual patient. This article provides eight design criteria to guide the ethical development and evaluation of step therapy policies and describes six clinical scenarios in which step therapy may be appropriate. The ethical criteria and scenarios are intended to provide guidance and transparency for insurers, patients, clinicians, and policy makers in choosing and paying for the appropriate therapies.


Effect of oral contraceptives on weight and body composition in young female runners.
To examine the effect of oral contraceptives (OC) on body weight, fat mass, percent body fat, and lean mass in young female distance runners. The study population consisted of 150 female competitive distance runners aged 18-26 yr who had participated in a 2-yr randomized trial of the effect of the OC Lo/Ovral (30 microg of ethinyl estradiol and 0.3 mg of norgestrel) on bone health. Weight and body composition were measured approximately yearly by balance beam scales and dual-energy x-ray absorptiometry, respectively. Women randomized to the OC group tended to gain slightly less weight (adjusted mean difference (AMD) = -0.54 +/- 0.31 kg.yr, P = 0.09) and less fat (AMD = -0.35 +/- 0.25 kg.yr, P = 0.16) than those randomized to the control group. OC assignment was associated with a significant gain in lean mass relative to controls among eumenorrheic women (those who had 10 or more menstrual cycles in the year before baseline; AMD = 0.77 +/- 0.17 kg.yr, P < 0.0001) but not among women with fewer than 10 menstrual cycles in that year (AMD = 0.02 +/- 0.35 kg.yr, P = 0.96). Treatment-received analyses yielded similar results. This randomized trial confirms previous findings that OC use does not cause weight or fat mass gain, at least among young female runners. Our finding that this OC is associated with lean mass gain in eumenorrheic runners, but not in those with irregular menses, warrants examination in other studies.


[Specific formation of esophageal autotransplant in the compromized and resected stomach].
An experience with 60 operated patients allows introduction of accuracy in the idea of possibility to use the compromised with different diseases and surgical procedures stomach in plasty of the esophagus. Technical specific features of forming the transplant are developed. Practicability of esophageal plasty with the small bowel using the loop of the bowel involved in digestive anastomoses is shown. In nonstandard situations the success of surgery is determined by an individual choice of plastic material, using reconstructive vascular surgery and rational method of placing the transplant.


Beyond the limitation of targeted therapy: Improve the application of targeted drugs combining genomic data with machine learning.
Precision oncology involves effectively selecting drugs for cancer patients and planning an effective treatment regimen. However, for Molecular targeted drug, using genomic state of the drug target to select drugs has limitations. Many patients who could benefit from molecularly targeted drugs, but they are being missed due to the insufficient labelling ability of the existing target genes. For non-specific chemotherapy drugs, most of the first-line anticancer drugs do not have biomarkers to guide doctor make treatment regimen. Furthermore, it is important to determine a long-term treatment plan based on the patient's genomic data during tumor evolution. Therefore, it is necessary to establish a tumor drug sensitivity prediction model, which can assist doctors in designing a personalized tumor treatment regimen. This paper proposed a novel model to predict tumor drug sensitivity including targeted drugs and non-specific chemotherapy drugs. This model uses statistical methods based on Bimodal distribution to select multimodal genetic data to solve dimensional challenges and reduce noise and to establish a classification model to predict the effectiveness of the drug in the tumor cell line using machine learning. The experimental test 87 molecular targeted drugs and non-specific chemotherapy drugs. The results show that the method can effectively predict the sensitivity of tumor drugs with an average sensitivity of 0.98 and specificity of 0.97. This model is worth to promotion. If it can be successfully used in clinical trials, it will effectively assist doctors to develop personalized cancer treatment programs and expand the application of molecularly targeted drugs.


Promoter hypermethylation of the p16 gene is associated with poor prognosis in recurrent early-stage hepatocellular carcinoma.
Despite significant advances in the detection and treatment of hepatocellular carcinoma, the prognosis of patients with hepatocellular carcinoma remains very poor, in part due to the high incidence of recurrence. This study was aimed at identifying a prognostic indicator of recurrence in patients with hepatocellular carcinoma. We retrospectively analyzed CpG island hypermethylation of the p14, p15, p16, GSTP1, integrin alpha4, SYK, and CDH1 genes in fresh-frozen tissues from 265 patients with hepatocellular carcinoma using the methylation-specific PCR. The expression levels of p16 and p53 were evaluated by immunohistochemistry. CpG island hypermethylation was detected in 6% for p14, 21% for p15, 67% for p16, 75% for GSTP1, 23% for integrin alpha4, 12% for SYK, and 57% for CDH1. Recurrence was observed in 102 (38%) of the 265 patients. There was no association between the risk for recurrence and hypermethylation of any gene studied. However, p16 methylation was associated with a poor survival after surgery for recurrent stage I to II hepatocellular carcinomas (hazard ratio, 4.05; 95% confidence interval, 1.15-14.20; P = 0.03). In addition, the hazard of failure after recurrence was about 3.80 (95% confidence interval, 1.03-14.20; P = 0.04) times higher in patients with p16 methylation than in those without. Negative expression of p16 at a protein level was also associated with poor survival in recurrent stage I to II hepatocellular carcinomas, but p53 expression did not have a synergistic effect on the poor prognosis. In conclusion, the present study suggests that p16 methylation may be associated with a poor prognosis in recurrent early-stage hepatocellular carcinomas.


The Political Determinants of China'S New Health Constitution.
The Basic Healthcare and Health Promotion Law 2019 became the new constitution of China's health system in June 2020, giving legal effect to ambitious health reform programmes like Healthy China 2030. The concurrent outbreak of coronavirus disease 2019 must not distract us from appreciating the fact that this Law will comprehensively overhaul the health regulatory framework of the world's most populous country during the coming decade, if not beyond. This article offers an original evaluation of the Law in its political context. The Law commendably promises to safeguard the right to health, assist citizens to live a 'complete cycle of life', and promote health using the resources of the public health system. However, it is also deeply politicised, guaranteeing extensive and penetrative political control in health campaigns, digitalised health data, the governance of health institutions, and the resolution of medical disputes. This can be explained by the consequential roles played by epidemics in China's historical dynastic cycles, but even more so by powerful tendencies of centralisation on the part of the Leninist Party-state. The Law's potential is thus subject to the overriding caveat that the Party-state's existence and influence over law and public health must be secured.


Charting pathways to climate change mitigation in a coupled socio-climate model.
Geophysical models of climate change are becoming increasingly sophisticated, yet less effort is devoted to modelling the human systems causing climate change and how the two systems are coupled. Here, we develop a simple socio-climate model by coupling an Earth system model to a social dynamics model. We treat social processes endogenously-emerging from rules governing how individuals learn socially and how social norms develop-as well as being influenced by climate change and mitigation costs. Our goal is to gain qualitative insights into scenarios of potential socio-climate dynamics and to illustrate how such models can generate new research questions. We find that the social learning rate is strongly influential, to the point that variation of its value within empirically plausible ranges changes the peak global temperature anomaly by more than 1°C. Conversely, social norms reinforce majority behaviour and therefore may not provide help when we most need it because they suppress the early spread of mitigative behaviour. Finally, exploring the model's parameter space for mitigation cost and social learning suggests optimal intervention pathways for climate change mitigation. We find that prioritising an increase in social learning as a first step, followed by a reduction in mitigation costs provides the most efficient route to a reduced peak temperature anomaly. We conclude that socio-climate models should be included in the ensemble of models used to project climate change.


H3 acetylation selectively promotes basal progenitor proliferation and neocortex expansion.
Increase in the size of human neocortex―acquired in evolution―accounts for the unique cognitive capacity of humans. This expansion reflects the evolutionarily enhanced proliferative ability of basal progenitors (BPs), including the basal radial glia and basal intermediate progenitors (bIPs) in mammalian cortex, which may have been acquired through epigenetic alterations in BPs. However, how the epigenome in BPs differs across species is not known. Here, we report that histone H3 acetylation is a key epigenetic regulation in bIP amplification and cortical expansion. Through epigenetic profiling of sorted bIPs, we show that histone H3 lysine 9 acetylation (H3K9ac) is low in murine bIPs and high in human bIPs. Elevated H3K9ac preferentially increases bIP proliferation, increasing the size and folding of the normally smooth mouse neocortex. H3K9ac drives bIP amplification by increasing expression of the evolutionarily regulated gene, Trnp1, in developing cortex. Our findings demonstrate a previously unknown mechanism that controls cortical architecture.


Excited-State Potential Energy Surfaces, Conical Intersections, and Analytical Gradients from Ground-State Density Functional Theory.
Kohn-Sham density functional theory (KS-DFT) has been a well-established theoretical foundation for ground-state electronic structure and has achieved great success in practical calculations. Recently, utilizing the eigenvalues from KS or generalized KS (GKS) calculations as an approximation to the quasiparticle energies, our group demonstrated a method to calculate the excitation energies from (G)KS calculation on the ground-state ( N - 1)-electron system. This method is now called QE-DFT (quasiparticle energies from DFT). In this work, we extend this QE-DFT method to describe excited-state potential energy surfaces (PESs), conical intersections, and the analytical gradients of excited-state PESs. The analytical gradients were applied to perform geometry optimization for excited states. In conjunction with several commonly used density functional approximations, QE-DFT can yield PESs in the vicinity of the equilibrium structure with accuracy similar to that from time-dependent DFT (TD-DFT). Furthermore, it describes conical intersection well, in contrast to TD-DFT. Good results for geometry optimization, especially bond length, of low-lying excitations for 14 small molecules are presented. The capability of describing excited-state PESs, conical intersections, and analytical gradients from QE-DFT and its efficiency based on just ground-state DFT calculations should be of great interest for describing photochemical and photophysical processes in complex systems.


Mexican Hat Modulation of Visual Acuity Following an Exogenous Cue.
Classical models of exogenous attention suggest that attentional enhancement at the focus of attention degrades gradually with distance from the attended location. On the other hand, the Attentional Attraction Field (AAF) model (Baruch and Yeshurun, 2014) suggests that the shift of receptive fields toward the attended location, reported by several physiological studies, leads to a decreased density of RFs at the attentional surrounds and hence the model predicts that the modulation of performance by spatial attention may have the shape of a Mexican Hat. Motivated by these theories, this study presents behavioral evidence in support of a Mexican Hat shaped modulation in exogenous spatial tasks that appears only at short latencies. In two experiments participants had to decide the location of a small gap in a target circle that was preceded by a non-informative attention capturing cue. The distance between cue and target and the latency between their onsets were varied. At short SOAs the performance curves were cubic and only at longer SOAs- this trend turned linear. Our results suggest that a rapid Mexican Hat modulation is an inherent property of the mechanism underlying exogenous attention and that a monotonically degrading trend, such as advocated by classical models, develops only at later stages of processing. The involvements of bottom-up processes such as the attraction of RFs to the focus of attention are further discussed.


Temporary overdriving pacing as an adjunct to antiarrhythmic drug therapy for electrical storm in acute myocardial infarction.
A-55-year-old man with diabetes mellitus was admitted to hospital because of chest pain. He was diagnosed as anterior acute myocardial infarction and treated with stent placement. After 7 days, ventricular fibrillation occurred because of a subacute reocclusion and balloon angioplasty was performed. Despite reperfusion therapy, intraaortic balloon pumping, antiarrhythmic drugs and beta-blocker, ventricular tachycardia or fibrillation relapsed and cardioversion was performed 29 times during 32 h. Temporary overdrive atrioventricular sequential pacing was initiated and the malignant arrhythmia finally disappeared. Even after stoppage of 25 h overdride pacing, it never recurred. Temporary overdrive pacing is an easy and feasible therapy for a drug-resistant electrical storm associated with AMI and should be performed in the early stage.


Movement patterns in rugby sevens: effects of tournament level, fatigue and substitute players.
Understanding of the physical demands and the effects of fatigue and substitute players in rugby sevens is limited. This study quantified the differences in movement patterns between domestic and international rugby sevens tournaments, the effects of fatigue within and between matches during tournaments, and movement patterns of second half substitute players. Movement patterns of 19 international-level male rugby sevens players were recorded using a Global Positioning System (GPS) device during 11 international and 16 domestic matches (n = 174 files). Maximum velocity, total distance covered, distance covered in velocity zones and number of moderate and high accelerations and decelerations are reported per min of match time. Movement patterns were compared between international and domestic matches, first and second half, first and last tournament match and substitute and full-match players. Substantially greater distance was covered at high velocity (~27% at ≥ 6 ms(-1)) and 4-39% more accelerations and decelerations were performed in international than domestic matches. The relative distance covered by players at velocities >2 ms(-1) and the number of changes in velocity were reduced by 1-16% from first to second half. Small differences were observed in activity at <5 ms(-1) (-8-8%) and moderate accelerations (-18%) from first to last tournament match. All movement variables were higher (2-123%) for substitute players. International rugby sevens competition is more intense than domestic matches. Despite reductions in work-rate within individual matches, there is little indication of accumulated fatigue over a multi-day tournament.


Antiphospholipid antibodies and thrombosis: association with acquired activated protein C resistance in venous thrombosis and with hyperhomocysteinemia in arterial thrombosis.
Although antiphospholipid antibodies (aPL) are associated with thrombosis, it is not known who with aPL is at higher risk for thrombosis. It was the aim of this cross-sectional study to investigate how thrombophilic factors contribute to venous or arterial thrombosis in aPL-positive individuals. In outpatient test centres at two tertiary care hospitals, two hundred and eight (208) persons requiring aPL testing were matched by age, gender and centre to 208 persons requiring a complete blood count. Persons were classified as aPL-positive (having anticardiolipin, lupus anticoagulant and/or anti-beta(2)-glycoprotein I antibodies) or aPL-negative. Several thrombophilic factors were studied using logistic regression modelling. Results showed that the aPL-positive group had three-fold more events (37%) than the aPL-negative group (12%). In unadjusted analyses, clinically important associations were observed between factor V Leiden and venous thrombosis, hyperhomocysteinemia and arterial thrombosis, and activated protein C resistance (APCR) and venous thrombosis (OR, 95% CI = 4.00, 1.35-11.91; 4.79, 2.03-11.33; and 2.03, 1.03-3.97, respectively). After adjusting for recruitment group, persons with both APCR and aPL had a three-fold greater risk (OR, 95% CI = 3.31, 1.30-8.41) for venous thrombosis than those with neither APCR nor aPL. Similarly, after adjusting for hypertension, family history of cardiovascular disease, gender and recruitment group, persons with both hyperhomocysteinemia and aPL had a five-fold increased risk (OR, 95% CI = 4.90, 1.37-17.37) for arterial thrombosis compared to those with neither risk factor. In conclusion, APCR phenotype and hyperhomocysteinemia are associated with a higher risk of venous and arterial thrombosis, respectively, in the presence of aPL.


Microstimulation mapping of precentral cortex during trained movements.
1. The precentral cortex of three Macaca mulatta monkeys were mapped with intracortical microstimulation (ICMS) while the monkeys performed alternating wrist flexion and extension movements. A forearm cocontraction task was also employed with one monkey. Electromyogram (EMG) recordings from forearm muscles were used to evaluate the results of ICMS. 2. We have found that the results of ICMS can be misleading if EMG activity is not recorded from the responding muscle. Inhibition can be interpreted as excitation if muscle palpation or joint movement are the only response criteria. 3. Movement of the stimulating electrode by as little as 200 microns in a single radial column sometimes changed EMG responses from inhibition to excitation or vice versa, indicating that cortical inhibitory areas for a muscle can be located very close to excitatory zones. 4. Both excitation and inhibition of muscles could be produced with ICMS of precentral cortex when the animal was performing a task involving the muscles being mapped. EMG responses to ICMS were stable, provided that the stimulation was applied at the same time during a repetitive task such that the motoneurons were at a given level of excitability. 5. Zones where ICMS produced inhibition of a particular forearm muscle were interspersed among zones that produced excitation for that muscle. 6. Regions exist in precentral cortex where ICMS activates antagonistic wrist muscles producing cocontraction. 7. The extensive cortical region from which any individual muscle can be activated or suppressed with ICMS and the various combinations of muscles that are activated from within this region suggest that different types of movements involving a single muscle are represented at different locations within this region. 8. At a few locations in precentral cortex, the EMG responses to ICMS were not just a function of the level of excitation of the motoneuron pool at the time of stimulation but were also dependent on the specific task the monkey was performing at the time of stimulation.


Comprehensive in Vitro Analysis of Acyltransferase Domain Exchanges in Modular Polyketide Synthases and Its Application for Short-Chain Ketone Production.
Type I modular polyketide synthases (PKSs) are polymerases that utilize acyl-CoAs as substrates. Each polyketide elongation reaction is catalyzed by a set of protein domains called a module. Each module usually contains an acyltransferase (AT) domain, which determines the specific acyl-CoA incorporated into each condensation reaction. Although a successful exchange of individual AT domains can lead to the biosynthesis of a large variety of novel compounds, hybrid PKS modules often show significantly decreased activities. Using monomodular PKSs as models, we have systematically analyzed the segments of AT domains and associated linkers in AT exchanges in vitro and have identified the boundaries within a module that can be used to exchange AT domains while maintaining protein stability and enzyme activity. Importantly, the optimized domain boundary is highly conserved, which facilitates AT domain replacements in most type I PKS modules. To further demonstrate the utility of the optimized AT domain boundary, we have constructed hybrid PKSs to produce industrially important short-chain ketones. Our in vitro and in vivo analysis demonstrated production of predicted ketones without significant loss of activities of the hybrid enzymes. These results greatly enhance the mechanistic understanding of PKS modules and prove the benefit of using engineered PKSs as a synthetic biology tool for chemical production.


[A cholesterol embolism following coronary angioplasty. A case report].
Cholesterol embolism is a rare but serious complication of heart catheterization. We report a patient in whom cholesterol embolization syndrome developed after coronary angioplasty complicated by an acute myocardial infarction which was treated with streptokinase and heparin. The clinical outcome was satisfactory. Cholesterol embolism occurrence might have been precipitated in this patient by thrombolytic and anticoagulant therapy.


Vasospasm secondary to ruptured aneurysm: assessment by digital intravenous angiography.
Digital intravenous angiography was used for the documentation and evaluation of cerebral vasospasm secondary to acute ruptured aneurysm. Attenuation of vessel caliber and/or generalized poor visualization of intracranial arteries and prolongation of circulation time were observed in seven patients with the acute clinical diagnosis of cerebral vasospasm. Posttreatment intravenous angiography demonstrated improvement in both arterial caliber and circulation time in six patients who responded to medical therapy for vasospasm. Digital intravenous angiography allowed repeat investigations of the intracranial vasculature safely and quickly without undue risk to the critically ill patient.


Stop codon in the procollagen II gene (COL2A1) in a family with the Stickler syndrome (arthro-ophthalmopathy).
Linkage analysis with restriction fragment length polymorphisms for the gene for type II procollagen (COL2A1) was carried out in a family with the Stickler syndrome, or arthro-ophthalmopathy, an autosomal dominant disorder that affects the eyes, ears, joints, and skeleton. The analysis demonstrated linkage of the disease and COL2A1 with a logarithm-of-odds score of 1.51 at zero recombination. A newly developed procedure for preparing cosmid clones was employed to isolate the allele for type II procollagen that was linked to the disease. Analysis of over 7000 nucleotides of the gene revealed a single base mutation that altered a CG dinucleotide and converted the codon CGA for arginine at amino acid position alpha 1-732 to TGA, a stop codon. From previous work on procollagen biosynthesis, it is apparent that the truncated polypeptide synthesized from an allele with a stop codon at alpha 1-732 cannot participate in the assembly of type II procollagen, and therefore that the mutation would decrease synthesis of type II procollagen. It was not apparent, however, why the mutation produced marked changes in the eye, which contains only small amounts of type II collagen, but relatively mild effects on the many cartilaginous structures of the body that are rich in the same protein.


Poor WASH (Water, Sanitation, and Hygiene) Conditions Are Associated with Leprosy in North Gondar, Ethiopia.
Access to safe water, sanitation, and hygiene (WASH) is critical for preventing the spread of neglected tropical diseases (NTDs) including leprosy. WASH-related transmission factors remain largely unexplored in the leprosy transmission cycle. The aim of this project is to better understand WASH exposures among leprosy cases through a case-control study in North Gondar, Ethiopia. We hypothesized that leprosy cases were more likely to have inadequate WASH access and were more likely to have concurrent schistosomiasis, as schistosomiasis immune consequences may facilitate leprosy infection. Forty leprosy cases (forty-one controls) were enrolled, tested for Schistosomamansoni, administered a demographic and WASH survey, and assigned a WASH index score. WASH factors significantly associated with leprosy on adjusted analyses included open defecation (aOR = 19.9, 95% CI 2.2, 176.3) and lack of access to soap (aOR = 7.3, 95% CI 1.1, 49.9). S. mansoni was detected in 26% of participants and in stratified analysis those with leprosy had a 3.6 (95% CI (0.8, 15.9)) greater odds of schistosomiasis in districts bordering the lake, compared to 0.33 lower odds of schistosomiasis in districts not bordering the lake (95% CI (0.09, 1.2)). Overall, results suggest that leprosy transmission may be related to WASH adequacy and access as well as to schistosomiasis co-infection.


Circumsolar sky radiation and turbidity of the atmosphere.
A statistical treatment of field measurements carried out by the Astrophysical Observatory of the Smithsonian Institution is presented. The brightness of a band of sky 10 degrees wide, concentric with the sun, has been determined. Measurements were obtained at two high altitude stations (Table Mountain and Montezuma) with the aid of a pyranometer constructed by C. G. Abbot and L. B. Aldrich. The primary object was to obtain an idea of the integral scattering of the sun's radiation by the atmospheric aerosol. Results have been used to determine long-periodic changes in the scattering properties of the atmosphere and their relation to other phenomena.


The collaboration of Antoine and Marie-Anne Lavoisier and the first measurements of human oxygen consumption.
Antoine Lavoisier (1743-1794) was one of the most eminent scientists of the late 18th century. He is often referred to as the father of chemistry, in part because of his book Elementary Treatise on Chemistry. In addition he was a major figure in respiratory physiology, being the first person to recognize the true nature of oxygen, elucidating the similarities between respiration and combustion, and making the first measurements of human oxygen consumption under various conditions. Less well known are the contributions made by his wife, Marie-Anne Lavoisier. However, she was responsible for drawings of the experiments on oxygen consumption when the French revolution was imminent. These are of great interest because written descriptions are not available. Possible interpretations of the experiments are given here. In addition, her translations from English to French of papers by Priestley and others were critical in Lavoisier's demolition of the erroneous phlogiston theory. She also provided the engravings for her husband's textbook, thus documenting the extensive new equipment that he developed. In addition she undertook editorial work, for example in preparing his posthumous memoirs. The scientific collaboration of this husband-wife team is perhaps unique among the giants of respiratory physiology.


A neural mechanism for contextualizing fragmented inputs during naturalistic vision.
With every glimpse of our eyes, we sample only a small and incomplete fragment of the visual world, which needs to be contextualized and integrated into a coherent scene representation. Here we show that the visual system achieves this contextualization by exploiting spatial schemata, that is our knowledge about the composition of natural scenes. We measured fMRI and EEG responses to incomplete scene fragments and used representational similarity analysis to reconstruct their cortical representations in space and time. We observed a sorting of representations according to the fragments' place within the scene schema, which occurred during perceptual analysis in the occipital place area and within the first 200 ms of vision. This schema-based coding operates flexibly across visual features (as measured by a deep neural network model) and different types of environments (indoor and outdoor scenes). This flexibility highlights the mechanism's ability to efficiently organize incoming information under dynamic real-world conditions.


Experimental and Clinical Evidence Supports the Use of Urokinase Plasminogen Activation System Components as Clinically Relevant Biomarkers in Gastroesophageal Adenocarcinoma.
Gastric and oesophageal cancers (GOCs) are lethal cancers which metastasise early and recur frequently, even after definitive surgery. The urokinase plasminogen activator system (uPAS) is strongly implicated in the invasion and metastasis of many aggressive tumours including GOCs. Urokinase plasminogen activator (uPA) interaction with its receptor, urokinase plasminogen activator receptor (uPAR), leads to proteolytic activation of plasminogen to plasmin, a broad-spectrum protease which enables tumour cell invasion and dissemination to distant sites. uPA, uPAR and the plasminogen activator inhibitor type 1 (PAI-1) are overexpressed in some GOCs. Accumulating evidence points to a causal role of activated receptor tyrosine kinase pathways enhancing uPAS expression in GOCs. Expression of these components are associated with poorer clinicopathological features and patient survival. Stromal cells, including tumour-associated macrophages and myofibroblasts, also express the key uPAS proteins, supporting the argument of stromal involvement in GOC progression and adverse effect on patient survival. uPAS proteins can be detected on circulating leucocytes, circulating tumour cells and within the serum; all have the potential to be developed into circulating biomarkers of GOC. Herein, we review the experimental and clinical evidence supporting uPAS expression as clinical biomarker in GOC, with the goal of developing targeted therapeutics against the uPAS.


ARL3 and ARL13B GTPases participate in distinct steps of INPP5E targeting to the ciliary membrane.
INPP5E, a phosphoinositide 5-phosphatase, localizes on the ciliary membrane via its C-terminal prenyl moiety, and maintains the distinct ciliary phosphoinositide composition. The ARL3 GTPase contributes to the ciliary membrane localization of INPP5E by stimulating the release of PDE6D bound to prenylated INPP5E. Another GTPase, ARL13B, which is localized on the ciliary membrane, contributes to the ciliary membrane retention of INPP5E by directly binding to its ciliary targeting sequence. However, as ARL13B was shown to act as a guanine nucleotide exchange factor (GEF) for ARL3, it is also possible that ARL13B indirectly mediates the ciliary INPP5E localization via activating ARL3. We here show that INPP5E is delocalized from cilia in both ARL3-knockout (KO) and ARL13B-KO cells. However, some of the abnormal phenotypes were different between these KO cells, while others were found to be common, indicating the parallel roles of ARL3 and ARL13B, at least concerning some cellular functions. For several variants of ARL13B, their ability to interact with INPP5E, rather than their ability as an ARL3-GEF, was associated with whether they could rescue the ciliary localization of INPP5E in ARL13B-KO cells. These observations together indicate that ARL13B determines the ciliary localization of INPP5E, mainly by its direct binding to INPP5E.


Outcomes of minimally invasive surgery for patients with endometrial carcinoma involving the cervix.
Most studies evaluating the oncologic safety of minimally invasive surgery for endometrial cancer focus on patients with stage I disease. The aim of this study was to investigate the outcomes of minimally invasive surgery for patients with endometrial carcinoma involving the cervix. Patients diagnosed between January 2010 and December 2015, with clinical stage II endometrial carcinoma, who underwent hysterectomy with lymphadenectomy, were drawn from the National Cancer Database. Inclusion criteria were clinical International Federation of Gynecology and Obstetrics (FIGO 2009) stage II, patients who underwent hysterectomy with lymphadenectomy, and known route of surgery (open or minimally invasive). Patients who received radiation therapy prior to surgery, those who had subtotal/supracervical hysterectomy, or unknown type of hysterectomy were excluded. The exposure of interest was performance of minimally invasive surgery either laparoscopic or robotic-assisted. Overall survival (primary endpoint) was assessed for patients diagnosed between January 2010 and December 2014 following generation of Kaplan-Meier curves and compared with the log-rank test. A Cox model was constructed to control for confounders. A total of 2175 patients were identified and 1282 (58.9%) had minimally invasive surgery. Of these, 339 and 943 patients had laparoscopic or robotic-assisted laparoscopic hysterectomy, respectively. Minimally invasive surgery was converted to open surgery in 74 (5.8%) patients. Those undergoing minimally invasive surgery had shorter hospital stay (median 1 vs 3 days, p<0.001), lower unplanned readmission rate (2.7% vs 4.7%, p=0.014), and 90-day mortality (0.8% vs 1.8%, p=0.05). Patients who had open surgery (n=796) had worse overall survival compared with those who had minimally invasive surgery (n=1048, p=0.003); 3-year overall survival rates were 76.8% and 83.6%, respectively. After controlling for patient age, race, type of insurance, presence of co-morbidities, performance of extensive lymphadenectomy, presence of positive lymph nodes, tumor histology, presence of lymphovascular space invasion, tumor size, and administration of radiotherapy, performance of minimally invasive surgery was not associated with worse survival (HR 0.90, 95% CI 0.73 to 1.11). In this retrospective analysis, minimally invasive surgery in patients with stage II endometrial carcinoma was associated with superior short-term peri-operative outcomes and improved 3-year overall survival.


Systemic alendronate treatment improves fixation of press-fit implants: a canine study using nonloaded implants.
Bone resorption associated with local trauma occurring during insertion of joint prostheses is recognized as an early event. Being an osteoclastic inhibitor, alendronate is a potential candidate means to decrease early periprosthetic bone resorption and thereby improve implant fixation. We investigated the influence of oral alendronate treatment on early implant fixation in two implant interface settings representing sites of an implant that are in contact with surrounding bone, and other sites without intimate bone contact. One plasma-sprayed cylindrical titanium implant (6 mm diameter) was inserted into each proximal tibia of 16 dogs. On one side the implant was inserted press-fit whereas on the contralateral side, the implants were surrounded by a 2 mm concentric gap. Oral alendronate (0.5 mg/kg/day) was given 2 weeks following surgery to eight dogs. The dogs were euthanized after 10 weeks of alendronate treatment. Bone ongrowth (bone in contact with implant surface) was estimated using the linear intercept technique and shear strength was calculated as the slope on a load-displacement curve. For the press fit implants, alendronate treatment significantly increased bone ongrowth from 24% to 29% and significantly increased ultimate shear strength from 1.26 to 3.72 MPa. Also, the fraction of periprosthetic bone significantly increased from 10% to 18%. For implants surrounded by a gap, alendronate neither stimulated nor impaired implant fixation, bone ingrowth, or new bone formation in the gaps. Because early implant stability is an important predictor of longevity, systemic alendronate treatment could be an important clinical tool to positively influence the early stages of implant incorporation.


Negative impact of Interleukin-9 on synovial regulatory T cells in rheumatoid arthritis.
In Rheumatoid Arthritis (RA), regulatory T cells (Tregs) have been found to be enriched in the synovial fluid. Despite their accumulation, they are unable to suppress synovial inflammation. Recently, we showed the synovial enrichment of interleukin-9 (IL-9) producing helper T cells and its positive correlation with disease activity. Therefore, we investigated the impact of IL-9 on synovial Tregs in RA. Here, we confirmed high synovial Tregs in RA patients, however these cells were functionally impaired in terms of suppressive cytokine production (IL-10 and TGF-β). Abrogating IL-9/ IL-9 receptor interaction could restore the suppressive cytokine production of synovial Tregs and reduce the synovial inflammatory T cells producing IFN-γ, TNF-α, IL-17. However, blocking these inflammatory cytokines failed to show any effect on IL-9 producing T cells, highlighting IL-9's hierarchy in the inflammatory network. Thus, we propose that blocking IL-9 might dampen synovial inflammation by restoring Tregs function and inhibiting inflammatory T cells.


Material Property Recovery by Controlling the Melt Memory Effects on Recrystallization and on Crystal Deformation: An Approach by the Molecular Dynamics Simulation for Polyethylene.
Degradation in the mechanical properties of recycled polymer materials has been recently appearing as a big issue in polymer science. The molecular mechanism of the degradation is considered in part due to residual memories of flow experienced during molding processes, and therefore the mechanical recycling through remolding involving melting and recrystallization has been attempted in recent years. In the present paper, the molecular processes of melting and recrystallization are investigated by the molecular dynamics simulation for polyethylene with special interest in the melt memory effects. We also studied the mechanical properties of the recrystalized materials that have undergone different recrystallization processes aiming to discover better recycling strategies. A successive step-by-step approach is adopted to study the loss of the crystal memory during retention in the melt, the effects of the melt memory on the mode of recrystallization, the relation between the recrystallization mode and the resulting higher-order structure, and the mechanical properties controlled by the higher-order structures. It is shown that the melt memory clearly remains in various order parameters that persist over time scales corresponding to the Rouse time, the remaining melt memory markedly affects the crystallization mode leading to distinct crystalline morphologies, and the distinct morphologies obtained give different mechanical responses during large deformations.


Morphology and spectral absorption characteristics of retinal photoreceptors in the southern hemisphere lamprey (Geotria australis).
The morphology and spectral absorption characteristics of the retinal photoreceptors in the southern hemisphere lamprey Geotria australis (Agnatha) were studied using light and electron microscopy and microspectrophotometry. The retinae of both downstream and upstream migrants of Geotria contained two types of cone photoreceptor and one type of rod photoreceptor. Visual pigments contained in the outer segments of these three photoreceptor types had absorbance spectra typical of porphyropsins and with wavelengths of maximum absorbance (downstream/upstream) at 610/616 nm (long-wavelength-sensitive cone, LWS), 515/515 nm (medium-wavelength-sensitive cone, MWS), and 506/500 nm (medium-wavelength-sensitive rod). A "yellow" photostable pigment was present in the myoid region of all three types of photoreceptor in the downstream migrant. The same short-wavelength-absorbing pigment, which prevents photostimulation of the beta band of the visual pigment in the outer segment, was present in the rods and LWS cones of the upstream migrant, but was replaced by a large transparent ellipsosome in the MWS cones. Using microspectrophotometric and anatomical data, the quantal spectral sensitivity of each photoreceptor type was calculated. Our results provide the first evidence of a jawless vertebrate, represented today solely by the lampreys and hagfishes, with two morphologically and physiologically distinct types of cone photoreceptors, in addition to a rod-like photoreceptor containing a colored filter (a cone-like characteristic). In contrast, all other lampreys studied thus far have either (1) one type of cone and one type of rod, or (2) a single type of rod-like photoreceptor. The evolution or retention of a second type of cone in adult Geotria is presumably an adaptation to life in the brightly lit surface waters of the Southern Ocean, where this species lives during the marine phase of its life cycle. The functional significance of the unique visual system of Geotria is discussed in relation to its life cycle and the potential for color vision.


Rhodanobacter rhizosphaerae sp. nov., isolated from soil of ginseng rhizosphere.
A Gram-stain-negative, aerobic, non-motile, non-spore-forming, yellow and rod-shaped bacterium, designated strain CR164T, was isolated from the rhizosphere soil of a ginseng field at Geumsan in Korea. CR164T grew at between 15 and 37 °C (optimal growth at 28 °C), between pH 6.0 and 9.0 (optimal growth at pH 7.0) and at salinities of 0-1.0 % (w/v) NaCl, growing optimally in the absence of NaCl. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that CR164T represents a member of the genus Rhodanobacter, showing the highest sequence similarity to Rhodanobactercaeni MJ01T (98.5 %), Rhodanobacter ginsenosidimutans Gsoil 3054T (98.4 %), Rhodanobacter thiooxydans LCS2T (98.3 %), Rhodanobacter lindaniclasticus RP5557T (98.1 %), Rhodanobacter denitrificans 2APBS1T (98.0 %), Rhodanobacter fulvus Jip2T (97.6 %), Rhodanobacter soli DCY45T (97.3 %) and 'Rhodanobacterxiangquanii' BJQ-6 (97.0 %). The major fatty acids were iso-C17 : 1ω9c (21.8 %), iso-C15 : 0 (12.1 %), iso-C11 : 0 (11.9 %) and iso-C16 : 0 (11.1 %). The predominant ubiquinone was Q-8. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The G+C content of the genomic DNA was 62.3 mol%. DNA-DNA relatedness between CR164T and the type strains of eight other species of the genus ranged from 51 to 9 %. On the basis of the polyphasic analysis, CR164T represents a novel species of the genus Rhodanobacter, for which the name Rhodanobacter rhizosphaerae sp. nov. is proposed. The type strain is CR164T (=KACC 18699T=NBRC 111845T).


Mechanisms mediating the enhanced gene transcription of P2X3 receptor by calcitonin gene-related peptide in trigeminal sensory neurons.
The molecular mechanisms underlying migraine pain remain unclear and probably require sustained facilitation in pain-sensing P2X(3) receptors gated by extracellular ATP in nociceptive sensory neurons. The major migraine mediator calcitonin gene-related peptide (CGRP) is known to sensitize P2X(3) receptors to increase impulse flow to brainstem trigeminal nuclei. This process is mediated via changes in the expression and function of P2X(3) receptors initially through enhanced trafficking and, later, perhaps through augmented synthesis of P2X(3) receptors. To clarify the mechanisms responsible for CGRP-evoked long lasting alterations in P2X(3) receptors, we used as a model mouse trigeminal ganglion neurons in culture. CGRP activated Ca(2+)-calmodulin-dependent kinase II, which became localized to the perimembrane region and neuronal processes, a phenomenon already apparent after 30 min and accompanied by a parallel increase in cAMP-response element-binding protein (CREB) phosphorylation and nuclear translocation. These effects triggered increased P2X(3) receptor transcription and were prevented by expressing a dominant negative form of CREB. Increased P2X(3) receptor synthesis was partly mediated by endogenous brain-derived neurotrophic factor (BDNF) because of its block by anti-BDNF antibodies and mimicry by exogenous BDNF. Immunocytochemistry experiments indicated distinct subpopulations of BDNF- or CGRP-sensitive trigeminal neurons with only partial overlap. The present data indicate a novel mechanism for enhancing P2X(3) receptor expression and function in trigeminal sensory neurons by CGRP via CREB phosphorylation. BDNF was an intermediate to extend the sensitizing effect of CGRP also to CGRP-insensitive neurons. This combinatorial action could serve as a powerful process to amplify and prolong pain mediated by P2X(3) receptors.


Adipose-tissue phosphofructokinase. Rapid purification and regulation by phosphorylation in vitro.
A new procedure for the purification of phosphofructokinase using Blue Dextran-Sepharose is described. This allowed an approx. 1000-fold purification of phosphofructokinase from rat white and brown adipose tissue to be achieved in essentially a single step. The purified enzymes from both tissues were found to exhibit hyperbolic kinetics with fructose 6-phosphate, to be inhibited by ATP and citrate, and to be activated by 5'-AMP, phosphate and fructose 2,6-bisphosphate. The enzymes were phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase, and phosphorylation was found to be associated with increases in activity when the enzymes were assayed under appropriate sub-optimal conditions. In particular, the phosphorylated enzymes exhibited less inhibition by ATP and the white-adipose-tissue enzyme was more sensitive to activation by fructose 2,6-bisphosphate. It is suggested that an increase in the cytoplasmic concentration of cyclic AMP in tissues other than liver may result in an increase in glycolysis through the phosphorylation of phosphofructokinase by cyclic AMP-dependent protein kinase.


Extraction of prostatic lumina and automated recognition for prostatic calculus image using PCA-SVM.
Identification of prostatic calculi is an important basis for determining the tissue origin. Computation-assistant diagnosis of prostatic calculi may have promising potential but is currently still less studied. We studied the extraction of prostatic lumina and automated recognition for calculus images. Extraction of lumina from prostate histology images was based on local entropy and Otsu threshold recognition using PCA-SVM and based on the texture features of prostatic calculus. The SVM classifier showed an average time 0.1432 second, an average training accuracy of 100%, an average test accuracy of 93.12%, a sensitivity of 87.74%, and a specificity of 94.82%. We concluded that the algorithm, based on texture features and PCA-SVM, can recognize the concentric structure and visualized features easily. Therefore, this method is effective for the automated recognition of prostatic calculi.


Cooking procedures for direct consumption of whole soybeans.
The purpose of this research was to determine adequate conditions for the preparation of soybeans at home level. In accordance with the results, soaking the soybeans for eight hours in a 0.25% bicarbonate solution, gives the appropriate hydration to the soybeans, which can then be easily cooked. Boiling them at atmospheric pressure during 20 minutes was required to destroy 80% of the soybeans antitrypsin activity, and a high PER was also obtained. Nevertheless, the soybeans texture was not satisfactory; therefore, longer boiling periods were tested so as to establish the most suitable time needed to achieve a softness similar to that of common beans as usually consumed. Softness was measured instrumentally, and the optimal conditions found were: 40 min boiling after 8 hr soaking in a 0.25% bicarbonate solution. Under these conditions, the PER and NPU values increased 18% and 33%, respectively, with respect to those of the casein PER and NPU. It is concluded that with this procedure, large amounts of protein can become available at an equivalent price to US$0.40/kg ($80.00/kg).


Age excuses: conversational management of memory failures in older adults.
The social consequences of appealing to age to excuse memory failure were examined in 2 vignette-based studies. In Study 1, 75 older (M = 72 years) and 78 young (M = 22 years) adults evaluated forgetful older targets in their 70s who used their age, lack of ability, lack of effort, or the situation to explain forgetting. In Study 2, 105 older (M = 72 years) and 105 young participants (M = 19 years) evaluated forgetful targets with no specific age given in 4 excuse conditions (age, ability, situation, and no excuse). In support of the prediction of positive consequences, age excuses were rated as more believable than situation in both studies and more believable and socially fluent than effort in Study 1. In support of predictions of negative consequences, both groups in Study 2 rated target persons who used an age excuse to be much older than their peers and, along with ability excuse users, as eliciting more worry and frustration than the others. Moreover, young adults showed additional sensitivity to the negative aspects of age excuses in terms of worry and frustration in Study 1 and anticipated repeat forgetting in Study 2. These results suggest that although age excuses may relieve socially awkward situations, this strategy reinforces negative age stereotyping of the older forgetter.


Expression profile of Wilms Tumor 1 (WT1) isoforms in undifferentiated and all-trans retinoic acid differentiated neuroblastoma cells.
Wilms tumor 1 gene (WT1) is a tumor suppressor gene originally identified in nephroblastoma. It is also expressed in neuroblastoma which represents the most aggressive extracranial pediatric tumor. Many evidences have shown that neuroblastoma may undergo maturation, by transforming itself in a more differentiated tumors such as ganglioneuroblastoma and ganglioneuroma, or progressing into a highly aggressive metastatic malignancy. To date, 13 WT1 mRNA alternative splice variants have been identified. However, most of the studies have focused their attention only on isoform of ∼49 kDa. In the present study, it has been investigated the expression pattern of WT1 isoforms in an in vitro model of neuroblastoma consisting in undifferentiated or all-trans retinoic acid (RA) differentiated cells. These latter representing the less malignant phenotype of this tumor. Results have demonstrated that WT1.1-WT1.5, WT1.6-WT1.9, WT1.10 WT1.11-WT1.12 and WT1.13 isoforms are expressed in both groups of cells, but their levels are significantly increased after RA treatment. These data have also been confirmed by immunofluorescence analysis. Moreover, the inhibition of PI3K/Akt and MAPK/ERK, that represent two signalling pathway specifically involved in NB differentiation, induces an overexpression of WT1 isoforms. These data suggest that WT1 isoforms might be involved in differentiation of neuroblastic into mature ganglion cells.