Advanced development of biomarkers for immunotherapy in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the most common liver cancer and one of the leading causes of cancer-related deaths in the world. Mono-immunotherapy and combination therapy with immune checkpoint inhibitors (ICIs) and multitargeted tyrosine kinase inhibitors (TKIs) or anti-vascular endothelial growth factor (anti-VEGF) inhibitors have become new standard therapies in advanced HCC (aHCC). However, the clinical benefit of these treatments is still limited. Thus, proper biomarkers which can predict treatment response to immunotherapy to maximize clinical benefit while sparing unnecessary toxicity are urgently needed. Contrary to other malignancies, up until now, no acknowledged biomarkers are available to predict resistance or response to immunotherapy for HCC patients. Furthermore, biomarkers, which are established in other cancer types, such as programmed death ligand 1 (PD-L1) expression and tumor mutational burden (TMB), have no stable predictive effect in HCC. Thus, plenty of research focusing on biomarkers for HCC is under exploration. In this review, we summarize the predictive and prognostic biomarkers as well as the potential predictive mechanism in order to guide future research direction for biomarker exploration and clinical treatment options in HCC.


Control of cerebral circulation in the neonate and infant.
Although much information is known about the factors controlling cerebral blood flow in the adult, there are significant physiological differences between the neonate, infant, and the adult. Therefore, it is not possible directly to transfer information concerning control of cerebral blood flow in the adult to the pediatric age population. A review of age-related differences in critical values of arterial oxygen tension (PaO2), arterial carbon dioxide tension (PaCO2), systemic arterial pressure and cerebral perfusion pressure (CCP) confirms that it is necessary to establish critical values in neonates and infants for each of these variables, as well as for any possible sympathetic nervous system influences on cerebral blood flow.


LHRH cells migrate on peripherin fibers in embryonic olfactory explant cultures: an in vitro model for neurophilic neuronal migration.
Luteinizing hormone releasing hormone (LHRH) neurons arise from progenitor cells in the olfactory placode. During prenatal development, these cells migrate via neurophilic interactions, in track-like arrangements along axons of the olfactory complex. The mechanisms by which these cells attain an adult-like distribution are unknown. In this study, we established an in vitro, embryonic mouse olfactory explant model to examine the factors directing LHRH cell migration. Cultures were generated from E11.5 embryos and maintained for up to 3 weeks. Typically 20-50% (160-400 LHRH cells) of the total LHRH neuronal population survived and maintained gene expression in these explants. Fibronectin and laminin staining delineated substratum producing cells which concentrically spread, from Days 1-7, from the periphery of the entire explanted tissue. In contrast, LHRH cells emigrated exclusively from inside olfactory pit areas to the surface of the culture after 3 days. The relationship between groups of LHRH cells emerging from bilateral olfactory pits was not random, but highly organized; in 93% of the cultures examined, the angular relationship between these groups was 180-270 degrees. After 5 or more days in vitro, LHRH cells were found in directional tracks similar to those observed in vivo forming a continuum from the olfactory pit out onto the substratum, where many bipolar LHRH cells were discretely located. The maximum distance away from the olfactory pit that LHRH cells were detected was 0.9 mm, which is compatible with the distance traversed by LHRH cells through the nasal region in vivo. The reproducible spatiotemporal appearance of LHRH cells was unrelated to the concentric spread of the fibronectin and laminin producing cells of the explant. Taken together, these data indicate that LHRH cells migrated directionally in these explants and that the molecular cues governing the initial migration of these neurons remained intact in this system in the absence of brain tissue. Double-label immunocytochemistry indicated that at least three populations of neuronal fibers existed in the explants: N-CAM-positive, peripherin-positive, and N-CAM/peripherin-positive. Although all three fibers groups showed highly organized spatiotemporal distribution patterns, only peripherin-positive fibers correlated with the location of LHRH cells. LHRH cells were always preceded by, and in close association with, peripherin-positive fibers. We hypothesize that signals arising from the peripherin-positive axons provide the appropriate guidance cues to LHRH cells as they emigrate from the olfactory pit.


Does Sex Influence Outcomes in Myocardial Infarction With Nonobstructive Coronary Arteries?
We examined differences in clinical profiles, predictors, and outcomes among patients with myocardial infarction (MI) with nonobstructive coronary arteries (MINOCAs) by sex. Data of 259 (132 males and 127 females) patients with MINOCA were consecutively collected. The primary clinical end point was major adverse cardiovascular events (MACE), including cardiovascular death, nonfatal MI, stroke, heart failure, and angina rehospitalization. Female patients with MINOCA were likely to be older than male patients with higher non-ST elevation myocardial infarction rate. Total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels were higher in female patients while male patients were more likely to have a smoking history, greater ST elevation myocardial infarction rate, higher diastolic blood pressure, and more alcohol use. During the 2-year follow-up, the incidence of MACE in males and females was similar (18% vs 20.2%, respectively; P = .673). The multivariable predictors of MACE in the female group were age, hypertension, and left ventricular ejection fraction (LVEF), whereas diabetes, smoking, and LVEF were multivariable predictors of MACE in the male group. In conclusion, there were differences in the clinical profiles between sexes. Clinical outcome was similar between male and female patients with MINOCA, whereas predictive risk factors varied.


Robot-Assisted Autism Therapy (RAAT). Criteria and Types of Experiments Using Anthropomorphic and Zoomorphic Robots. Review of the Research.
Supporting the development of a child with autism is a multi-profile therapeutic work on disturbed areas, especially understanding and linguistic expression used in social communication and development of social contacts. Previous studies show that it is possible to perform some therapy using a robot. This article is a synthesis review of the literature on research with the use of robots in the therapy of children with the diagnosis of early childhood autism. The review includes scientific journals from 2005-2021. Using descriptors: ASD (Autism Spectrum Disorders), Social robots, and Robot-based interventions, an analysis of available research in PubMed, Scopus and Web of Science was done. The results showed that a robot seems to be a great tool that encourages contact and involvement in joint activities. The review of the literature indicates the potential value of the use of robots in the therapy of people with autism as a facilitator in social contacts. Robot-Assisted Autism Therapy (RAAT) can encourage child to talk or do exercises. In the second aspect (prompting during a conversation), a robot encourages eye contact and suggests possible answers, e.g., during free conversation with a peer. In the third aspect (teaching, entertainment), the robot could play with autistic children in games supporting the development of joint attention. These types of games stimulate the development of motor skills and orientation in the body schema. In future work, a validation test would be desirable to check whether children with ASD are able to do the same with a real person by learning distrust and cheating the robot.


Multiplex fluorescent analysis of four short tandem repeats for rapid haemophilia A molecular diagnosis.
Indirect molecular diagnosis of hemophiliaA (HA) is carried out by analyzing intragenic polymorphic markers described along the coagulation factorVIII (FVIII) gene. Several studies have demonstrated that the two commonly used intronic short tandem repeats (STR13 and STR22) located in the FVIII gene are highly informative for this task. Two extragenic markers closely linked to FVIII (DXS1073 and DXS1108) have also been described as valuable tools for gene tracking. The objective of the present work was to develop a rapid, single-tube automated method to simultaneously analyze these four STRs. Consistent amplification was achieved by quadruplex fluorescent PCR and the products were analyzed by capillary electrophoresis. Validation of the method included DNA analysis of 88 individuals from a control population, 45 HA patients and 32 individuals from 10 HA-affected families. Statistical study showed that the STR13, STR22 and DXS1108 loci were in significant linkage disequilibrium, whereas DXS1073 was not. Nevertheless, the combination of the four markers offered a high heterozygosity rate (>90%) that improved tracing of FVIII gene inheritance. Optimal results with application to single cells in a HA preimplantation genetic diagnosis (PGD) protocol demonstrated the sensitivity of the technique. In conclusion, the automated fluorescent method described is an extremely rapid, simple and highly informative one that is easy to standardize and allows direct comparison of results among different groups working with genetic counseling, prenatal diagnosis and PGD in HA-affected families.


Atypical bipolar II depression compared with atypical unipolar depression and nonatypical bipolar II depression.
Aim of the study was to find out whether atypical bipolar II depression was distinct from both atypical unipolar depression and nonatypical bipolar II depression. Seventy-nine consecutive atypical bipolar II depressed outpatients were compared with 42 consecutive atypical unipolar depressed outpatients and with 53 consecutive nonatypical bipolar II depressed outpatients. Among the variables studied (age at intake, age at onset, female gender, duration of illness, psychosis, comorbidity, chronicity, recurrences, severity), age at intake and onset were significantly lower in the atypical bipolar II group than in the other groups. The other variables, apart from psychosis, were not significantly different. Findings suggest that atypical bipolar II depression may have an age at onset different from that of atypical unipolar depression and nonatypical bipolar II depression. As different ages at onset may identify distinct subtypes of depression, this finding might suggest that atypical bipolar II depression may be distinct from both atypical unipolar depression and nonatypical bipolar II depression.


[Operative strategy for redo heart valve operation].
Thirty-five redo heart valve operations were performed between Jan. 1990 and Dec. 1993 in Jichi Ohmiya Medical Center. Previous operations were mitral valve replacement (MVR) in 19, closed mitral commissurotomy in 1, open mitral commissurotomy in 6, aortic valve replacement (AVR) in 5, ASD repair in 3, VSD repair in 1. Thirty (85.7%) were in NYHA functional class III or less, and 5 (14.3%) were in functional class in IV. Twenty-three patients had undergone MVR, 5 AVR, 4 MVR+AVR, 2 TVR (TAP), 1 Cabrol operation, and 8 patients had undergone concomitant tricuspid valve repair. Emergent operation was none. Overall operative mortality was 5.7% (2/35), as the cause of death was low output syndrome. Chest CT was available for the evaluation of the distension of right atrium and right ventricle. Catastrophic hemorrhage by the injury of ascending aorta or right atrium during redo sternotomy was 4, but that was not the cause of death. We emphasize the necessity of the exposure of the femoral artery and the preparation for cardiopulmonary bypass before sternotomy. The extent of dissecting adhesion should be determined case by case respectively under consideration of the risk of cardiovascular injury and postoperative large bleeding. Furthermore the cannule of SVC or IVC with cuff was useful for the cases with strong adhesion.


COMPARING THE OUTCOME OF SINGLE VERSUS MULTIPLE SESSION LASER PHOTOABLATION OF FLAT NEOVASCULARIZATION IN ZONE 1 AGGRESSIVE POSTERIOR RETINOPATHY OF PREMATURITY: A Prospective Randomized Study.
To compare single versus 2-session laser photoablation for flat neovascularization in cases with Zone 1 aggressive posterior retinopathy of prematurity. Twenty-nine Asian Indian infants with aggressive posterior retinopathy of prematurity were randomized; each eye received 1 of 2 methods (29 each in Group A or B) proposed by the PHOTO-ROP group. Group A underwent single session laser to the avascular retina underlying the flat neovascularization by direct laser over the fronds. Group B underwent laser in 2 sessions; first, laser was delivered to the avascular periphery up to the flat neovascularization and 7 days later to the avascular bed exposed by the retraction of the fronds. Outcome and complications between the two groups were compared. Mean birthweight and gestational ages were 1,276 g and 30.1 weeks, respectively. All eyes showed favorable outcome at a minimum 12-month follow-up. Hemorrhages after laser (41.4% vs. 17.2%, P < 0.001) were more common in the single laser group. Large hemorrhages (>1 disk diameter) seen in Group A took longer than 8 weeks to resolve and developed focal fibrosis. This study demonstrates that the two-staged laser procedure produces fewer and smaller hemorrhages and no fibrosis compared with a single session. Both methods have comparable favorable outcomes in Asian Indian infants.


[Immunohistochemical diagnosis of metastasizing prostatic carcinomas].
Metastases of 47 known prostatic carcinomas were subjected to the unlabelled immunoperoxidase-procedure to localise prostaticacid-phosphatase (PAP) and prostatic-specific antigen (PSA). In bone-marrow, lymph-node, lung and liver metastases PAP was found in 64% and PSA in 78%. There was no significant difference between the intensity of staining in primary and metastatic neoplasm. In poorly differentiated metastases of prostatic adenocarcinomas less intense staining for PAP and PSA was found. The data suggest that the demonstration of PAP and PSA is a practical and sensitive test for the prostatic origin of a clinically and histologically unclassifiable metastasis.


Leiomyomas of the lower respiratory tract: a case report.
A rare case of leiomyoma of the left main bronchus in a 16-year-old female has been reported. Of the pulmonary leiomyomas reported, 18.3 per cent were found in the trachea, 33.8 per cent in the bronchi and 47.9 per cent in the parenchyma. Bronchial lesions tend to produce symptoms due to partial or complete obstruction of the affected bronchus, whereas, over 90 per cent of parenchymal lesions which are more common in women, are asymptomatic and usually discovered on routine chest radiographs. Tracheal lesions may present as bronchial asthma. The correct diagnosis is not usually made until the lesion has been resected. However, early and correct diagnosis may be made in suspicious cases by histological examination of bronchoscopic biopsy specimens or frozen section material obtained at thoracotomy. The surgical approach to these tumors is conservative but lobectomy or pneumonectomy is often necessary because of secondary pulmonary destruction.


Blocking ER export of the Golgi SNARE SYP31 affects plant growth.
We recently identified a novel and transplantable di-acidic motif (EXXD) that facilitates ER export of the Golgi syntaxin SYP31 (type IV protein) and which may function also for type I and type II proteins in plants. By mutagenesis of Arabidopsis thaliana SYP31 and live cell imaging experiments in tobacco leaf epidermal cells, we determined that replacing the MELAD sequence of SYP31 with gagag retained SYP31 in the ER, which demonstrates that the di-acidic motif ELAD is critical for SYP31 ER export. To investigate whether blockage of a Golgi SNARE in the ER have consequences for plant growth, we produced tobacco plants stably overexpressing either the wild type MELAD or the mutant gagag form of SYP31. Whereas tobacco plants overexpressing the wild-type SYP31 developed to set seed, tobacco plants overexpressing the mutant form gagag rapidly became chlorotic, ceased their growth and invariably died after several weeks. This indicated that retention of overexpressed SYP31 in the ER is likely toxic for the secretory pathway and, therefore, plant development. Putative explanations for this observation are discussed taking into account SNARE properties and possible interactions.


Research on depression in Parkinson disease: A bibliometric and visual analysis of studies published during 2012-2021.
The diagnosis and treatment rate of Parkinson disease (PD) with depression has a low diagnostic rate, and there is no consensus on the choice of treatment mode. This study evaluates the global research trends of scientific outputs related to depression in PD from multiple perspectives, using a bibliometric analysis and visualization tool to scientifically analyze the knowledge from the literature. Literature related to depression in PD published from 2012 to 2021 was included and selected from the Web of Science Core Collection database in October 2021. CiteSpace software was used to visualize and analyze co-occurrence analyses for countries, institutions, authors, and keywords. A total of 4533 articles from the Web of Science database were included. The United States made the largest contribution with the majority of publications (1215; 29.40%). Toronto University was the most productive institution. PD, depression, quality of life, dementia, nonmotor symptom, prevalence, anxiety, Alzheimer disease, symptom, and disorder would be significantly correlated with depression in PD. The current hot spots in this field focus on the following: risk factors for depression in PD, assessment scale of depression in PD, and rehabilitation of depression in PD. This analysis not only reveals the current research trends and hotspots but also provides some instructive suggestions on the development of depression in PD.


Recent advances in i-Gene tools and analysis: microarrays, next generation sequencing and mass spectrometry.
Recent advances in technology and associated methodology have made the current period one of the most exciting in molecular biology and medicine. Underlying these is an appreciation that modern research is driven by increasing large amounts of data being interpreted by interdisciplinary collaborative teams which are often geographically dispersed. The availability of cheap computing power, high speed informatics networks and high quality analysis software has been essential to this as has the application of modern quality assurance methodologies. In this review, we discuss the application of modern 'High-Throughput' molecular biological technologies such as 'Microarrays' and 'Next Generation Sequencing' to scientific and biomedical research as we have observed. Furthermore in this review, we also offer some guidance that enables the reader as to understand certain features of these as well as new strategies and help them to apply these i-Gene tools in their endeavours successfully. Collectively, we term this 'i-Gene Analysis'. We also offer predictions as to the developments that are anticipated in the near and more distant future.


Moral issues in workplace health promotion.
There is debate to what extent employers are entitled to interfere with the lifestyle and health of their workers. In this context, little information is available on the opinion of employees. Within the framework of a workplace health promotion (WHP) program, moral considerations among workers were investigated. Employees from five companies were invited to participate in a WHP program. Both participants (n = 513) and non-participants (n = 205) in the program filled in a questionnaire on individual characteristics, lifestyle, health, and opinions regarding WHP. Nineteen percent of the non-participants did not participate in the WHP program because they prefer to arrange it themselves, and 13% (also) preferred to keep private life and work separate. More participants (87%) than non-participants (77%) agreed with the statement that it is good that employers try to improve employees' health (χ(2) = 12.78, p = 0.002), and 26% of the non-participants and 21% of the participants think employer interference with their health is a violation of their privacy. Employees aged 50 year and older were more likely to agree with the latter statement than younger workers (OR = 1.56, 95% CI 1.02-2.39). This study showed that most employees support the importance of WHP, but in a modest group of employees, moral considerations may play a role in their decision whether or not to participate in WHP. Older workers were more likely to resist employer interference with their health. Therefore, special attention on such moral considerations may be needed in the communication, design, and implementation of workplace health promotion programs.


New intraventricular catheter for volume pressure response measurements.
Measurements of the volume-pressure response (VPR) to determine the relative position of a patient on the graphic volume-pressure curve have been used to derive clinically useful information. One reason that these measurements have not been used more frequently has been the fear of introducing infection into the ventricular system. We have designed an intraventricular catheter that allows repeated VPR measurements and reduces the risk of infection.


Model studies on volatile release from different semisolid fat blends correlated with changes in sensory perception.
The effect of dispersed aqueous droplets in water-in-oil (W/O)-emulsion semisolid fats on aroma release and sensory perception was investigated on margarine models where model aroma substances were added. Aroma release from W/O-emulsion fat blends and bulk fat blends with added monoglycerides combining different fatty acids of various short-chain free fatty acids, methylketones, esters, and lactones were measured using headspace solid phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS), and their perception profiles were evaluated by sensory analysis. The presence of aqueous phase in a fat blend significantly reduced the headspace concentrations of butanoic acid and hexanoic acid, and also decreased the perceived intensity of total aroma and cheesy aroma. The aroma release of methylketones, esters, and lactones from the W/O-emulsion fat blends increased with increasing carbon chain length of the volatile molecules. The intensity of aroma perception in a W/O-emulsion fat blend depended on the melting point of the fatty acids (oleic, palmitic, stearic, and behenic) of the monoglyceride used as an emulsifier. Thus, aroma release from a W/O-emulsion semisolid fat blend was influenced by interactions between aroma volatiles and the dispersed aqueous droplets and by their viscoelastic properties.


Prediction of N-myristoylation modification of proteins by SVM.
Attachment of a myristoyl group to NH(2)-terminus of a nascent protein among protein post-translational modification (PTM) is called myristoylation. The myristate moiety of proteins plays an important role for their biological functions, such as regulation of membrane binding (HIV-1 Gag) and enzyme activity (AMPK). Several predictors based on protein sequences alone are hitherto proposed. However, they produce a great number of false positive and false negative predictions; or they cannot be used for general purpose (i.e., taxon-specific); or threshold values of the decision rule of predictors need to be selected with cautiousness. Here, we present novel and taxon-free predictors based on protein primary structure. To identify myristoylated proteins accurately, we employ a widely used machinelearning algorithm, support vector machine (SVM). A series of SVM predictors are developed in the present study where various scales representing physicochemical and biological properties of amino acids (from the AAindex database) are used for numerical transformation of protein sequences. Of the predictors, the top ten achieve accuracies of >98% (the average value is 98.34%), and also the area under the ROC curve (AUC) values of >0.98. Compared with those of previous studies, the prediction accuracies are improved by about 3 to 4%.


The potential role of small fish species in improving micronutrient deficiencies in developing countries: building evidence.
To build a comprehensive overview of the potential role of fish in improving nutrition with respect to certain micronutrient deficiencies in developing countries. A comprehensive literature review was completed. For this the electronic library databases ASFA, CABD and Scopus were systematically searched and relevant references cited in these sources were carefully analysed. The search terms used were 'fish', 'small fish species', 'micronutrients', 'food-based strategies', 'fish consumption' and 'developing countries'. The quality of data on nutritional analyses was carefully reviewed and data that lacked proper information on methods, units and samples were excluded. The evidence collected confirmed the high levels of vitamin A, Fe and Zn in some of the small fish species in developing countries. These small fish are reported to be more affordable and accessible than the larger fish and other usual animal-source foods and vegetables. Evidence suggests that these locally available small fish have considerable potential as cost-effective food-based strategies to enhance micronutrient intakes or as a complementary food for undernourished children. However, the present review shows that only a few studies have been able to rigorously assess the impact of fish consumption on improved nutritional status in developing countries. Further research is required in areas such as determination of fish consumption patterns of poor households, the nutritional value of local fish and other aquatic animals and the impact of fish intake on improved nutritional status in developing countries where undernutrition is a major public health problem.


Enzyme-Loaded Flower-Shaped Nanomaterials: A Versatile Platform with Biosensing, Biocatalytic, and Environmental Promise.
As a result of their unique structural and multifunctional characteristics, organic-inorganic hybrid nanoflowers (hNFs), a newly developed class of flower-like, well-structured and well-oriented materials has gained significant attention. The structural attributes along with the surface-engineered functional entities of hNFs, e.g., their size, shape, surface orientation, structural integrity, stability under reactive environments, enzyme stabilizing capability, and organic-inorganic ratio, all significantly contribute to and determine their applications. Although hNFs are still in their infancy and in the early stage of robust development, the recent hike in biotechnology at large and nanotechnology in particular is making hNFs a versatile platform for constructing enzyme-loaded/immobilized structures for different applications. For instance, detection- and sensing-based applications, environmental- and sustainability-based applications, and biocatalytic and biotransformation applications are of supreme interest. Considering the above points, herein we reviewed current advances in multifunctional hNFs, with particular emphasis on (1) critical factors, (2) different metal/non-metal-based synthesizing processes (i.e., (i) copper-based hNFs, (ii) calcium-based hNFs, (iii) manganese-based hNFs, (iv) zinc-based hNFs, (v) cobalt-based hNFs, (vi) iron-based hNFs, (vii) multi-metal-based hNFs, and (viii) non-metal-based hNFs), and (3) their applications. Moreover, the interfacial mechanism involved in hNF development is also discussed considering the following three critical points: (1) the combination of metal ions and organic matter, (2) petal formation, and (3) the generation of hNFs. In summary, the literature given herein could be used to engineer hNFs for multipurpose applications in the biosensing, biocatalysis, and other environmental sectors.


Essential role of citron kinase in cytokinesis of spermatogenic precursors.
During spermatogenesis, the first morphological indication of spermatogonia differentiation is incomplete cytokinesis, followed by the assembly of stable intercellular cytoplasmic communications. This distinctive feature of differentiating male germ cells has been highly conserved during evolution, suggesting that regulation of the cytokinesis endgame is a crucial aspect of spermatogenesis. However, the molecular mechanisms underlying testis-specific regulation of cytokinesis are still largely unknown. Citron kinase is a myotonin-related protein acting downstream of the GTPase Rho in cytokinesis control. We previously reported that Citron kinase knockout mice are affected by a complex neurological syndrome caused by cytokinesis block and apoptosis of specific neuronal precursors. In this report we show that, in addition, these mice display a dramatic testicular impairment, with embryonic and postnatal loss of undifferentiated germ cells and complete absence of mature spermatocytes. By contrast, the ovaries of mutant females appear essentially normal. Developmental analysis revealed that the cellular depletion observed in mutant testes is caused by increased apoptosis of undifferentiated and differentiating precursors. The same cells display a severe cytokinesis defect, resulting in the production of multinucleated cells and apoptosis. Our data indicate that Citron kinase is specifically required for cytokinesis of the male germ line.


Spermidine and spermine are enriched in whole blood of nona/centenarians.
Polyamines (putrescine, spermidine, and spermine) are a family of molecules that derive from ornithine through a decarboxylation process. They are essential for cell growth and proliferation, stabilization of negative charges of DNA, RNA transcription, translation, and apoptosis. Recently, it has been demonstrated that exogenously administered spermidine promotes longevity in yeasts, flies, worms, and human cultured immune cells. Here, using a cross-sectional observational study, we determined whole-blood polyamines levels from 78 sex-matched unrelated individuals divided into three age groups: Group 1 (31-56 years, n=26, mean age 44.6±6.07), group 2 (60-80 years, n=26, mean age 68.7±6.07), and group 3 (90-106 years, n=26, mean age 96.5±4.59). The total content of polyamines is significantly lower in groups 2 and 3 compared to group 1 (p=3.6×10(-12)). Interestingly, this reduction is mainly attributable to the lower putrescine content. Group 2 displays the lowest levels of spermidine and spermine. On the other hand, nona/centenarians (group 3) display a significantly higher median relative percentage content of spermine with respect to total polyamines, compared to the other groups (13.2% vs. 14.1% vs. 30.6%, p=6.0×10(-4)). For the first time, we report profiles of polyamines from the whole blood of healthy nona/centenarians, and our results confirm and extend previous findings on the role of polyamines in determining human longevity. However, although we found an important correlation between polyamines levels and age groups, further studies are warranted to fully understand the role of polyamines in determining life span. Also, longitudinal and nutritional studies might suggest potential therapeutic approaches to sustain healthy aging and to increase human life span.


[Mortality of patients with a history of acute myocardial infarct 3 months after the disease].
The lethality rate of patients with past myocardial infarction after the third month of the infarction is discussed. Two groups of patients with past myocardial infarction were followed up for 4 years after the third month of the infarction. The basic group included patients who carried out a rehabilitation program while the patients in the control group were not included in any kind of rehabilitation. At the of the first year the lethality rate of the basic group of patients was 5.71%% while of the control group it was 8.82%. In the second year the lethality rate fell in the basic group to 4.55% and in the control group to 6.45%. In the third year the lethality rate was 3.17% for the basic group and 3.45% for the control group, and in the fourth year it was 1.63% for the basic group and 3.57% for the control group. The rehabilitation program carried out with the patients of the basic group had exerted a favorable influence on the survival of the patients and had led to a lower total lethality rate of 15.06% compared to that of 22.29% of the control group for the whole four year period of observation. The physical rehabilitation reduces the known until now factors of increased risk as well as the possibility for a new infarction.


Modeling and Optimization for a New Compliant 2-dof Stage for Locating Biomaterial Samples by an Efficient Approach of a Kinetostatic Analysis-Based Method and Neural Network Algorithm.
The nanoindentation technique is employed to characterize the behaviors of biomaterials. Nevertheless, there is a lack of development of a miniaturized precise positioner for in situ nanoindentation. Besides, modeling behaviors of the positioner are restricted due to its complex kinematic characteristics. Therefore, this paper proposes a novel compliant two degrees of freedom (dof) stage for positioning a biomaterial sample in in situ nanoindentation. In addition, a new modeling and dimensional optimization synthesis method of the stage is developed. The proposed effective methodology is developed based on a kinetostatic analysis-based calculation method, the Lagrange approach, and a neural network algorithm. With an increased advance in artificial intelligence, a neural network algorithm is proposed to extend the applicability of artificial neural networks in optimizing the parameters of the stage. First, the 2-dof stage is built via a combination of an eight-lever displacement amplifier and a symmetric parallelogram mechanism. Second, a chain of mathematical equations of the 2-dof stage is constructed using a kinetostatic analysis-based method to calculate the ratio of displacement amplification and the input stiffness of the 2-dof stage. Then, the Lagrange method is utilized to formulate the dynamic equation of the 2-dof stage. Finally, a neural network algorithm is adopted to maximize the natural first frequency of the proposed stage. The optimal results determined that the frequency of the stage can achieve a high value of 112.0995 Hz. Besides, the formed mathematical models were relatively precise by comprising the simulation verifications.


Role of endothelin-1 in regulation of the postnatal intestinal circulation.
Newborn intestine is uniquely prone to vasoconstriction in response to a wide variety of perturbations. To test the hypothesis that endothelin (ET)-1 is an important factor in this process, we determined the effects of exogenous ET-1 administration and blockade of endogenous ET-1 in vivo and in vitro in 3- and 35-day-old swine. Intramesenteric artery administration of exogenous ET-1 to vascularly isolated in vivo gut loops (10(-9) M/kg bolus) caused vasoconstriction and reduced gut O(2) uptake similarly in these age groups. Selective blockade of ET(A) or ET(B) receptors with BQ-610 or BQ-788, respectively, in vascularly isolated in vivo gut loops had no effect on gut vascular resistance or O(2) uptake in either age group; within in vitro gut loops, BQ-610 significantly increased vasoconstriction when perfusion pressure was reduced below baseline, but only in 3-day-old animals; i.e., it impaired the autoregulatory response to perfusion pressure reduction. Exogenous ET-1 significantly decreased capillary perfusion within in vitro gut loops, as evidenced by a decrease in capillary filtration coefficient, but only in 3-day-old animals; furthermore, blockade of endogenous ET-1 activity with BQ-610 significantly enhanced capillary filtration coefficient in 3-day-old animals and increased O(2) extraction ratio. ET-1 did not depress intestinal metabolic rate, as evidenced by its effect on the O(2) uptake-blood flow relationship; it did compromise tissue oxygenation because of its effects on intestinal O(2) transport. ET-1 concentration in mesenteric venous effluent exceeded arterial concentration, but only in 3-day-old intestine, suggesting production of ET-1 by newborn intestine. We conclude that ET-1 exerts an age-dependent effect on intestinal hemodynamics in postnatal intestine, having a greater impact in 3- than in 35-day-old intestine.


A subspace approach based on embedded prewhitening for voice activity detection.
This paper presents a subspace approach for voice activity detection (VAD). The proposed approach is based on an embedded prewhitening scheme for the simultaneous diagonalization of the clean speech and noise covariance matrices to provide a decision rule based on likelihood ratio test in signal subspace domain. Experimental results show that the proposed subspace-based VAD algorithm outperforms the method using a Gaussian model in a conventional discrete Fourier transform domain at the low signal-to-noise conditions.


Membrane-perturbing properties of three peptides corresponding to the ectodomain of hepatitis C virus E2 envelope protein.
Based on the predicted capacity to interact with membranes at the interface, we have found three regions in the ectodomain of the hepatitis C virus envelope glycoprotein E2 (430-449, 543-560 and 603-624) with the ability to destabilize membranes. Three peptides corresponding to the sequence of these regions have been synthesized and their interaction with liposomes have been characterized. The three peptides were able to insert deeply into the hydrophobic core of negatively charged phospholipids as stated by fluorescence depolarization of the probe 1,6-diphenyl-1,3,5-hexatriene. Peptides E2(430-449) and E2(603-624) were able to induce aggregation of phosphatidylglycerol vesicles in a concentration-dependent manner both at neutral and acidic pH while peptide E2(543-560) did not induce any increase of optical density at 360 nm in the concentration range studied. The three peptides induced lipid mixing and the release of the internal contents in a dose-dependent manner when acidic phospholipids were used. Fourier transformed infrared spectroscopy indicated that the peptides adopted mainly a beta-sheet conformation which is not modified by the presence of acidic phospholipids. Taken together, our results point out to the involvement of these three regions in the fusion mechanism of HCV at the plasma membrane level.


Sb-Doped high-voltage LiCoO2 enabled improved structural stability and rate capability for high-performance Li-ion batteries.
Sb-Doped high-voltage LiCoO2 was developed with unique properties. In situ X-ray diffraction and density functional theory reveal that the introduction of Sb helps to shorten the Li+ diffusion distance, increase the lattice spacing and keep the structural stability during deep lithiation, thus resulting in improved rate capability and cycling performance.


[Human fast negative EEG potentials before express saccades].
Fast negative EEG potentials preceding fast regular saccades and express saccades were studied by the method of backward averaging under conditions of monocular stimulation of the right and left eye. "Step" and "gap" experimental paradigms were used for visual stimulation. Analysis of parameters of potentials and their spatiotemporal dynamics suggests that, under conditions of the increased attention and optimal readiness of the neural structures, express saccades appear when the previously chosen program of the future eye movement coincides with the actual target coordinates. We assumed that the saccade latency decreases at the expense of the involvement of the main oculomotor areas of motor and saccadic planning in its initiation; an express saccade can be initiated also by means of direct transmission of the signal from the cortex to the brainstem saccadic generator passing by the superior colliculus. Moreover, anticipating release from the central fixation and attention distraction are necessary for the successful initiation of an express saccade.


Rac Inhibition Causes Impaired GPVI Signalling in Human Platelets through GPVI Shedding and Reduction in PLCγ2 Phosphorylation.
Rac1 is a small Rho GTPase that is activated in platelets upon stimulation with various ligands, including collagen and thrombin, which are ligands for the glycoprotein VI (GPVI) receptor and the protease-activated receptors, respectively. Rac1-deficient murine platelets have impaired lamellipodia formation, aggregation, and reduced PLCγ2 activation, but not phosphorylation. The objective of our study is to investigate the role of Rac1 in GPVI-dependent human platelet activation and downstream signalling. Therefore, we used human platelets stimulated using GPVI agonists (collagen and collagen-related peptide) in the presence of the Rac1-specific inhibitor EHT1864 and analysed platelet activation, aggregation, spreading, protein phosphorylation, and GPVI clustering and shedding. We observed that in human platelets, the inhibition of Rac1 by EHT1864 had no significant effect on GPVI clustering on collagen fibres but decreased the ability of platelets to spread or aggregate in response to GPVI agonists. Additionally, in contrast to what was observed in murine Rac1-deficient platelets, EHT1864 enhanced GPVI shedding in platelets and reduced the phosphorylation levels of PLCγ2 following GPVI activation. In conclusion, Rac1 activity is required for both human and murine platelet activation in response to GPVI-ligands, but Rac1's mode of action differs between the two species.


Out-of-pocket expenditure for hospitalization in Haryana State of India: Extent, determinants & financial risk protection.
India aspires to achieve universal health coverage, which requires ensuring financial risk protection (FRP). This study was done to assess the extent of out-of-pocket (OOP) expenditure and FRP for hospitalization in Haryana State, India. Further, the determinants for FRP were also evaluated. Data collected as a part of a household level survey conducted in Haryana 'Concurrent Evaluation of National Rural Health Mission: Haryana Health Survey' were analyzed. Descriptive analysis was undertaken to assess socio-demographic characteristics, hospitalization rate, extent and determinants of OOP expenditure and FRP. Prevalence of catastrophic health expenditure (CHE) (more than 40% of non-food expenditure) and impoverishment (Int$ 1.25) were estimated. Multivariate logistic regression was used to assess determinants of FRP. Hospitalization rate was found to be 3106 persons or 3307 episodes per 100,000 population. Median OOP expenditure on hospitalization was ₹ 8000 (USD 133), which was predominantly attributed to medicines (37%). Prevalence of CHE was 25.2 per cent with higher prevalence amongst males [odds ratio (OR)=1.30], those belonging to scheduled caste and scheduled tribes (OR=1.35), poorest 20 per cent households (OR=3.05), having injuries (OR=4.03) and non-communicable diseases (OR=3.13) admitted in a private hospital (OR=2.69) and those who were insured (OR=1.74). There was a 12 per cent relative increase in poverty head count due to OOP payments on healthcare. Our findings showed that hospitalization resulted in significant OOP expenditure, leading to CHEs and impoverishment of households. Impact of OOP expenditures was inequitably more on the vulnerable groups. OOP expenditure may be curtailed through provision of free medicines and diagnostics and removal of any form of user charges.


Ultra-compact MXene fibers by continuous and controllable synergy of interfacial interactions and thermal drawing-induced stresses.
Recent advances in MXene (Ti3C2Tx) fibers, prepared from electrically conductive and mechanically strong MXene nanosheets, address the increasing demand of emerging yet promising electrode materials for the development of textile-based devices and beyond. However, to reveal the full potential of MXene fibers, reaching a balance between electrical conductivity and mechanical property is still the fundamental challenge, mainly due to the difficulties to further compact the loose MXene nanosheets. In this work, we demonstrate a continuous and controllable route to fabricate ultra-compact MXene fibers with an in-situ generated protective layer via the synergy of interfacial interactions and thermal drawing-induced stresses. The resulting ultra-compact MXene fibers with high orientation and low porosity exhibit not only excellent tensile strength and ultra-high toughness, but also high electrical conductivity. Then, we construct meter-scale MXene textiles using these ultra-compact fibers to achieve high-performance electromagnetic interference shielding and personal thermal management, accompanied by the high mechanical durability and stability even after multiple washing cycles. The demonstrated generic strategy can be applied to a broad range of nanostructured materials to construct functional fibers for large-scale applications in both space and daily lives.


Increased levels of vascular endothelial growth factor and advanced glycation end products in aqueous humor of patients with diabetic retinopathy.
Clinical studies have shown a relationship between diabetic retinopathy and vascular endothelial growth factor (VEGF) levels in ocular fluid. Advanced glycation end products (AGEs) have been implicated in diabetes complications, including diabetic retinopathy. Nepsilon-(carboxymethyl) lysine (CML) is a glycoxidation product that may be a marker of oxidative stress. In this study, we used enzyme-linked immunosorbent assays to determine the levels of VEGF, non-CML AGE and CML in the aqueous humor and serum of 82 Japanese patients with type 2 diabetes and 60 non-diabetic subjects. VEGF, non-CML AGE, and CML concentrations in aqueous humor and serum were then compared with the severity of diabetic retinopathy. Immunohistochemical detection analysis of non-CML AGE and CML was also performed using retinal tissues from patients with progressive diabetic retinopathy. Aqueous levels of VEGF, non-CML AGE and CML increased along with the progression of diabetic retinopathy compared to age-matched controls. After coagulation therapy, the VEGF, non-CML AGE, and CML levels were significantly reduced. Immunostaining showed diffuse co-localization of non-CML AGE and CML around microvessels and in the glial cells of proliferative membranes from patients with progressive diabetic retinopathy. These findings suggest that glycation and glycoxidation reactions (or oxidation, as revealed by CML) may contribute to both the onset and progression of diabetic retinopathy.


Rapid emotional response and disadvantageous Iowa gambling task performance in women with borderline personality disorder.
Adults with Borderline Personality Disorder (BPD) manifest poor performance on tasks of decision making which may be congruent with their decisional and interpersonal conflicts in real life. Poor decision making is often assumed to be due to impulsive behaviour or weak inhibitory control despite inconsistent evidences of these relationships, leaving questions about the specific nature of these decisional deficits. Decision making in BPD may be compromised by different domains of impulsivity, affective dysregulatory processes or unknown co-morbid ADHD which is considered a developmental precursor to BPD. Iowa Gambling Task (IGT) decision making, 2 tasks of inhibitory control and a self report of ADHD symptoms consisting of 9 subscales were administered to 41 BPD women and 41 healthy controls. No group differences in inhibitory control were present. Net decision making performance and all ADHD subscale ratings differed significantly among BPD women and healthy controls. BPD women did not meet the threshold indicative of moderate to severe ADHD. Three subscales of attention, behaviour/ disorganized and emotive were significantly associated with poor IGT performance in 26 women with BPD. Of these 3 variables, the emotive subscale, representing a rapid emotional response, was the only significant predictor contributing 49% to the variance in poor DM. This is the 1st evidence of an emotive type of impulsivity, representing a type of affective instability that is linked to poor IGT DM in BPD. Findings support the Somatic Marker Hypothesis of IGT DM and may reflect the affective dysregulation that characterizes the disorder.


Effects of hydrostatic pressure on the electron [Formula: see text] factor and g-factor anisotropy in GaAs-(Ga, Al)As quantum wells under magnetic fields.
The hydrostatic-pressure effects on the electron-effective Landé [Formula: see text] factor and g-factor anisotropy in semiconductor GaAs-Ga(1-x)Al(x)As quantum wells under magnetic fields are studied. The [Formula: see text] factor is computed by considering the non-parabolicity and anisotropy of the conduction band through the Ogg-McCombe effective Hamiltonian, and numerical results are displayed as functions of the applied hydrostatic pressure, magnetic fields, and quantum-well widths. Good agreement between theoretical results and experimental measurements in GaAs-(Ga, Al)As quantum wells for the electron g factor and g-factor anisotropy at low values of the applied magnetic field and in the absence of hydrostatic pressure is obtained. Present results open up new possibilities for manipulating the electron-effective g factor in semiconductor heterostructures.


Cell-mediated resistance to infection with Listeria monocytogenes in nude mice.
Congenitally dysthymic nude (nu/nu) NMRI mice showed increased resistance to viable Listeria monocytogenes cells during the initial phase of infection as compared with euthymic control mice. The intravenous mean lethal dose (LD50), as determined for euthymic mice after an observation time of 7 and 14 days, amounted consistently to 6 X 10(4) Listeria. The corresponding values determined in nude mice were found to be increased by either 20-fold (1.2 X 10(6) Listeria after an observation time of 7 days) or 4-fold (2.4 X 10(5) Listeria after an observation time of 14 days). The transfer of spleen cells from immune euthymic donor mice into chronically infected nude mice caused almost complete elimination of Listeria within 1 week. The injection of dextran sulfate 24 h before a secondary infection with L. monocytogenes caused loss of antibacterial resistance in both chronically infected nude mice and Listeria-immune euthymic mice, this being expressed by a rapid increase in the numbers of bacteria in the spleens as well as the occurrence of serious signs of illness.


The prevalence of appendiceal fecaliths in patients with and without appendicitis. A comparative study from Canada and South Africa.
Appendicitis is more common in developed than in developing societies and appendiceal fecaliths are thought to have an etiologic role in the disease. The geographic distribution of appendiceal fecaliths was investigated by systematic, intraoperative palpation of the appendix in patients in Toronto, Canada and Johannesburg, South Africa. The incidences of fecaliths found on pathologic sectioning of the appendix in appendicitis patients in both societies were compared. In the Canadian population, the prevalence of fecaliths in patients whose appendices were palpated incidentally was 32% versus 52% for those with appendicitis (p less than 0.01). In the African population, the prevalence of fecaliths in patients whose appendices were palpated incidentally was four per cent versus 23% for those with appendicitis (p = 0.04). The difference in prevalence of incidental appendiceal fecaliths in the two populations was statistically significant (p less than 0.005). The prevalence of fecaliths is higher in developed countries, such as Canada, than in developing countries, such as Africa, and is also higher in patients with than in those without appendicitis. These data support the theory that the low-fiber diets consumed in developed countries lead to fecalith formation, which then predisposes to appendicitis.


Immunoperoxidase demonstration of Chlamydia inclusions in routine Pap-smears.
Indirect immunoperoxidase with antichlamydia serum has been applied to 34 Pap-smears where cytologic diagnosis of Chlamydia infection was suspected. Only 4 cases were confirmed by this technique. The limited success could be due to the need for a more active antiserum or to an overestimation of Pap-smear images. Cytologic diagnostic features are critically outlined.


Mixed Mulberry Fruit and Mulberry Leaf Fermented Alcoholic Beverages: Assessment of Chemical Composition, Antioxidant Capacity In Vitro and Sensory Evaluation.
The fruit and leaves of mulberry (Morus alba L.) contain a variety of health-beneficial components. At present, although both alcoholic and non-alcoholic mulberry beverages are commercially available, studies on fermented alcoholic beverages mixed with both mulberry fruit and mulberry leaf are rare. In this study, different amounts (1, 2 and 3%, g/mL dry weight) of mulberry leaves were added during the alcoholic fermentation of mulberry juice. After 9 days of fermentation, the 1-deoxynojirimycin (DNJ) content increased from 61.12 ± 3.10 to 153.39 ± 3.98 μg/mL, and the quercetin content increased from 0.45 ± 0.01 to 20.14 ± 0.08 mg/L in the mulberry alcoholic beverages with the addition of mulberry leaves at 2%. Moreover, the ABTS+ scavenging capacity at the end of fermentation for the same sample was enhanced by 40.9%. In addition, the total sugar, total phenols, total anthocyanins, and γ-aminobutyric acid (GABA) contents of the fermented samples all decreased significantly at the end of fermentation. A total of 33 volatile compounds and 17 free amino acids were detected in the fermented alcoholic beverages with mulberry leaves added. The total free amino acid content increased with the increase in mulberry leaf addition. Principal component analysis showed that the addition of mulberry leaves during fermentation increased the contents of GABA, DNJ, total flavonols and protocatechuic acid in mulberry alcoholic beverages. All these studies revealed the dynamic changes in functional components in the alcoholic fermentation of mulberry juices with the addition of mulberry leaves. Overall, the addition of mulberry leaf powder at 2% was selected as the appropriate addition for producing mulberry alcoholic beverages with enhanced nutritional value.


A molecular mechanism for variations in muscle function in rainbow trout.
Salmonids undergo a developmental transition from parr to smolt that involves a number of physiological and morphological changes. In recent years, my laboratory has studied shifts in red muscle function at this parr-smolt transformation (PST) in rainbow trout, Oncorhynchus mykiss. Parr red muscle has faster contraction kinetics than smolts, including faster rates of activation and relaxation and a faster maximum shortening velocity. At PST, a transition in swimming behavior is also observed, with lower tailbeat frequencies and longer EMG duty cycles in the older smolts. Lastly, there is molecular correlate to changes in kinetics and behavior. During PST, there is a developmental reduction in the number of myosin heavy chain (MHC) isoforms in the red muscle of rainbow trout. Since MHC composition of muscle can determine contractile properties, these molecular results suggest a mechanism for the transition in red muscle kinetics and steady swimming. The red muscle of parr is more likely to contain the fast-twitch or white isoform of MHC, resulting in faster contractile properties of that muscle and higher tailbeat frequencies during steady swimming. Lastly, experimental work supports the conclusion that the shift in kinetics causes the observed shift in swimming behavior.


Morphologic changes of alveolar macrophages in smoking sarcoidosis patients.
In a group of 105 patients suffering from various pulmonary diseases we have previously observed quantitative and qualitative differences between alveolar macrophages from smokers and non-smokers. In this study, we compared the cell population of bronchoalveolar lavage (BAL) fluids in non-smoking, and smoking patients suffering from pulmonary sarcoidosis. Cell yield was significantly higher in smokers than in non-smokers: (P less than 0.02), lavage fluids of smokers contained 83% of macrophages and 14% of lymphocytes, as compared with 60% and 34% respectively in non-smokers (P less than 0.002). However, the total number of lymphocytes was not significantly different between the two groups. Alveolar macrophages from smokers were bigger and contained large pigmented lipoproteinic inclusions. Complementary cytochemical studies showed that lipofuscins were the principal constituent of these intracytoplasmic inclusions. In conclusion, cigarette smoke increases the number and alters the structure of the alveolar macrophages of patients with sarcoidosis, as well as in other chronic respiratory disease as previously shown. The mechanism responsible for the increased lipofuscins concentration in the alveolar macrophages of smokers remains to be determined.


Effects of elevated carbon dioxide concentration and temperature on needle growth, respiration and carbohydrate status in field-grown Scots pines during the needle expansion period.
We determined effects of long-term elevation of carbon dioxide concentration ([CO2]) and temperature on growth, respiration and carbohydrate concentration in needles of field-grown Scots pine (Pinus sylvestris L.) trees during the needle expansion period. Sixteen 20-year-old Scots pine trees were individually enclosed in closed-top, environmentally controlled chambers for 4 years in one of four environments: ambient conditions (CON); elevated [CO2] (EC); elevated temperature (ET); and a combination of both (EC + ET). Needle growth, carbohydrate concentration and dark respiration were measured at 3-day intervals throughout the needle expansion period. Dark respiration was partitioned into growth and maintenance components by regressing specific respiration rate against specific growth rate. In all treatments, growth, carbohydrate concentration and daily dark respiration rates of needles followed a similar seasonal pattern throughout the needle expansion period. Treatments EC, ET and EC + ET increased individual needle area and dry weight compared with the CON treatment. Carbohydrate concentrations in needles were increased by EC, but reduced by ET and EC + ET. Daily respiration rates increased slightly in the early stage of needle expansion and decreased gradually in the late stage when needles were exposed to EC, but increased consistently throughout the growing period when needles were exposed to ET or EC + ET. Partitioning of respiration into its two functional components showed that the growth respiration coefficient was unaffected by the treatments, whereas maintenance respiration was reduced by EC but increased by ET and EC + ET. Maintenance respiration was more sensitive to elevated temperature than growth respiration. We conclude that the difference in respiration rates between expanding and expanded needles should be taken into account when estimating the respiratory responses of needles to elevated [CO2] and temperature.


[Infectious endocarditis in non-addict patients without predisposing heart disease. Differential features].
Although uncommonly, infective endocarditis in non-addict patients may involve people without predisponente heart disease. The aim of our study was to assess the clinical and prognostic features of this type of endocarditis and to compare them with those of the more common type of endocarditis with underlying lesion. With this aim, we have reviewed 71 consecutive cases of non-addict infective endocarditis diagnosed in our hospital in the last 7 years; there was no preexisting cardiac lesion in 9 patients (13% of all endocarditis and 21% of native valve endocarditis), while underlying heart disease, including mitral valve prolapse, was present in the remaining 62 patients. Mean age was significantly lower in 9 patients without preexistent lesion (28 +/- 18 versus 46 +/- 17 years, p < 0.01), while there was no differences for gender. Infection involved the aortic valve in 56%, the tricuspid or pulmonary valve in 33% and the mitral valve in only 11% of the patients without underlying cardiopathy (for 44%, 4% and 49%, respectively, in patients with cardiopathy). Staphylococcus aureus caused 67% of cases in patients without cardiopathy and only 9% in those with cardiopathy. Surgery was required in a similar proportion by both groups of patients (55% and 56%), although mortality was more than twice higher in patients with prior cardiac lesions (25% versus 11%). A significant proportion of non-addict infective endocarditis involves patients without predisponente heart disease. These cases have some differential features (younger age, aortic and right heart valves involvement, S. aureus as the main causative agent and lower mortality) in comparison to those of endocarditis in patients with underlying cardiopathy.


Comparison of Superior-Level Facet Joint Violations Between Robot-Assisted Percutaneous Pedicle Screw Placement and Conventional Open Fluoroscopic-Guided Pedicle Screw Placement.
To compare the superior-level facet joint violations (FJV) between robot-assisted (RA) percutaneous pedicle screw placement and conventional open fluoroscopic-guided (FG) pedicle screw placement in a prospective cohort study. This was a prospective cohort study without randomization. One-hundred patients scheduled to undergo RA (n = 50) or FG (n = 50) transforaminal lumbar interbody fusion were included from February 2016 to May 2018. The grade of FJV, the distance between pedicle screws and the corresponding proximal facet joint, and intra-pedicle accuracy of the top screw were evaluated based on postoperative CT scan. Patient demographics, perioperative outcomes, and radiation exposure were recorded and compared. Perioperative outcomes include surgical time, intraoperative blood loss, postoperative length of stay, conversion, and revision surgeries. Of the 100 screws in the RA group, 4 violated the proximal facet joint, while 26 of 100 in the FG group had FJV (P = 0.000). In the RA group, 3 and 1 screws were classified as grade 1 and 2, respectively. Of the 26 FJV screws in the FG group, 17 screws were scored as grade 1, 6 screws were grade 2, and 3 screws were grade 3. Significantly more severe FJV were noted in the FG group than in the RA group (P = 0.000). There was a statistically significant difference between RA and FG for overall violation grade (0.05 vs 0.38, P = 0.000). The average distance of pedicle screws from facet joints in the RA group (4.16 ± 2.60 mm) was larger than that in the FG group (1.92 ± 1.55 mm; P = 0.000). For intra-pedicle accuracy, the rate of perfect screw position was greater in the RA group than in the FG group (85% vs 71%; P = 0.017). No statistically significant difference was found between the clinically acceptable screws between groups (P = 0.279). The radiation dose was higher in the FG group (30.3 ± 11.3 vs 65.3 ± 28.3 μSv; P = 0.000). The operative time in the RA group was significantly longer (184.7 ± 54.3 vs 117.8 ± 36.9 min; P = 0.000). Compared to the open FG technique, minimally invasive RA spine surgery was associated with fewer proximal facet joint violations, larger facet to screw distance, and higher intra-pedicle accuracy.


A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis.
Transcriptional coregulators are vital to the efficient transcriptional regulation of nuclear chromatin structure. Coregulators play a variety of roles in regulating transcription. These include the direct interaction with transcription factors, the covalent modification of histones and other proteins, and the occasional chromatin conformation alteration. Accordingly, establishing relatively quick methods for identifying proteins that interact within this network is crucial to enhancing our understanding of the underlying regulatory mechanisms. LC-MS/MS-mediated protein binding partner identification is a validated technique used to analyze protein-protein interactions. By immunoprecipitating a previously-identified member of a protein complex with an antibody (occasionally with an antibody for a tagged protein), it is possible to identify its unknown protein interactions via mass spectrometry analysis. Here, we present a method of protein preparation for the LC-MS/MS-mediated high-throughput identification of protein interactions involving nuclear cofactors and their binding partners. This method allows for a better understanding of the transcriptional regulatory mechanisms of the targeted nuclear factors.


Semipermanent capillary coatings in mixed organic-water solvents for CE.
This report describes the creation of semipermanent capillary coatings that are compatible with organic-water solvent systems in CE. The coatings are created by simply rinsing the fused-silica capillary with long double-chain cationic surfactants, such as dimethyl-ditetradecyl ammonium bromide (2C(14)DAB), dihexadecyldimethyl ammonium bromide (2C(16)DAB), and dimethyldioctadecyl ammonium bromide (2C(18)DAB). These surfactants generate semipermanent bilayer coatings on the capillary surface, which display a high degree of stability in buffers containing up to 60% v/v of organic solvents, such as methanol and ACN. The coating stability increases with increasing hydrophobicity of the surfactant, i.e., with increasing chain length. For instance, the EOF changes by only 1.2% in a 2C(18)DAB-coated capillary after 130 capillary volumes of rinsing with 60% v/v methanol containing buffer. The bilayer coatings allow separations to be performed without the need to regenerate the coating between runs or to maintain the EOF modifier in the run buffer. Rapid separations (<2 min) of anions and basic drugs with migration time reproducibility of less than 0.5% RSD and efficiencies of 0.4-0.6 million plates/m are obtained. In addition, selectivity changes for small anions and cationic drugs are also observed when the organic solvent content is adjusted.


Risk Assessment of (Herbal) Teas Containing Pyrrolizidine Alkaloids (PAs) Based on Margin of Exposure Approach and Relative Potency (REP) Factors.
Pyrrolizidine alkaloids (PAs) present distinct toxicity potencies depending on their metabolites and in vivo toxicokinetics. To represent the potency differences of various PAs, the interim relative potency (REP) factors have been derived. However, little is known about the risk assessment for (herbal) teas when taking REP factors into account. In this study, a set of 68 individual 1,2-unsaturated PA in 21 types of (herbal) teas was analyzed using LC-MS/MS. The REP factors for these PAs were applied on the PA levels. The margin of exposure (MOE) approach was employed to assess the risks of the exposure to PAs due to consumption of (herbal) teas. The results show that the total PA levels ranged from 13.4 to 286,682.2 μg/kg d.m., which were decreased by REP correction in most of the teas. The MOE values for tephroseris, borage and lemon balm (melissa) tea based on REP-corrected PA levels were below 10,000, assuming daily consumption of one cup of tea during a lifetime, indicating that consuming these teas may raise a concern. Our study also indicates a priority for risk management for tephroseris tea, as having nephrosis tea for more than 11.2 weeks during a 75-year lifetime would result in an MOE of 10,000.


[Surgical reconstruction of long esophageal stenoses in children].
Four cases related with the long stenosis of lower part of esophagus are referred to in children. These were stenoses of fibromuscular type, due to achalasia and resulting from etching with sulphuric acid as well as from tumor. The extramucous myotomy, esophagotomy and colonic replacement of long esophageal stenosis were elected. The condition is repairable by esophagomyotomy or esophagotomy when the function of stenotic portion of esophagus is preserved. To prevent postoperative complications such as fistula, diverticulum, gastroesophageal reflux and restenosis, the covering of incision with a patch from gastric fundus is recommended with subsequent fundoplication. Anatomical and functional lesion of esophageal wall in the presence of long stenosis requires its esophagoplasty as well as replacement with the intestinal interposition.


Technology-based interventions for psychiatric illnesses: improving care, one patient at a time.
Worldwide, individuals with severe psychiatric illnesses struggle to receive evidence-based care. While science has made remarkably slow progress in the development and implementation of effective psychiatric treatments, we have witnessed enormous progress in the emergence and global penetration of personal computing technology. The present paper examines how digital resources that are already widespread (e.g., smartphones, laptop computers), can be leveraged to support psychiatric care. These instruments and implementation strategies can increase patient access to evidenced-based care, help individuals overcome the barriers associated with the stigma of mental illness, and facilitate new treatment paradigms that harness wireless communication, sensors and the Internet, to enhance treatment potency. Innovative digital treatment programmes that have been used successfully with a range of conditions (i.e., schizophrenia, posttraumatic stress disorder and borderline personality disorder) are presented in the paper to demonstrate the utility and potential impact of technology-based interventions in the years ahead.


Detection of Atypical Salmonella Infantis Phenotypes in Broiler Environmental Samples.
In numerous countries, strict and targeted measures concerning Salmonella monitoring and control are implemented and high quality of surveillance is ensured by obligatory investigation of samples from the primary production level of animals according to EN/ISO standards. Here, 2 phenotypic characteristics of Salmonella exhibited on compulsory media are crucial, namely, motility demonstrated on modified semisolid Rappaport Vassiliadis agar (MSRV), and production of hydrogen sulfide (H2S) on xylose lysine deoxycholate agar (XLD). In the present study, we describe the detection of Salmonella Infantis variants found in broiler environmental samples with major alterations in their growth characteristics on MSRV, XLD, and brilliant green-phenol red-agar (BPLS). The variants proved to be non-motile on MSRV and displayed non-confirming colony appearances on the previously mentioned selective agars. The growth spectrum comprised pinhead sized yellow colonies with small black centers, but also pinpoint sized colorless colonies, both colony types of regular shape. Our work contributes to highlight the finding of S. Infantis variants which possess more than one phenotypic deviation from the "typical" growth characteristics and by this limit the detection power of the actual obligatory used media. IMPORTANCE Salmonellosis caused by non-typhoidal Salmonella serovars is the second most frequently reported zoonotic disease in humans in the EU. The transmission of these agents is mainly via contaminated food of animal origin. In this context, poultry products are the main source of infection. Therefore, continuous and standardized surveillance of the prevalence of such Salmonella serovars at the primary production level is essential. Our findings show the phenotypic heterogeneity of the serovar Infantis and provide growth characteristics of atypical variants. Such variants pass unnoticed official screening methods, resulting in incorrect identification and being underrepresented in epidemiological surveillance programs.


[The usefulness of two-dimensional transesophageal echocardiography in follow-up studies of dissecting aortic aneurysm].
Two-dimensional transesophageal echocardiography (2D-TEE) facilitates clear imaging of the thoracic aorta, and we have already reported the diagnostic value of this procedure for detecting dissecting aortic aneurysm (DAA). Recently, we performed preoperative and postoperative 2D-TEE in nine surgically treated patients with DAA (Debakey type I: 2 cases, type II: 1 case, type IIIa: 2 cases, type IIIb: 4 cases), and repeated 2D-TEE in six medically treated patients (DeBakey type IIIa: 2 cases, type IIIb: 4 cases), and the usefulness of 2D-TEE in the follow-up study of DAA was evaluated. Two types of ultrasound probe (radial scan type, and sector type with a color-Doppler system) were used in this study. Of the surgically treated patients, four of nine cases in which the initial tears were excised or repaired showed neither leakage nor pseudoaneurysm at the site of graft insertion. Other surgically treated patients showed residual intimal tears and continued to harbor patent false lumen. One patient with type I dissection demonstrated localized saccular aneurysm in the proximal arch of the aorta, but no evidence of aortic regurgitation. Of the six medically treated patients, further extension of the initial dissection was observed in one of the two type IIIa cases. False lumen narrowing was recognizable in three of the four type IIIb patients, and complete obliteration of false lumen was observed at necropsy in one of these three cases. Double-channeled aorta was seen in another case of type IIIb dissection.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of family and peers in the initiation and continuation of smoking behavior of future physicians.
Globally researchers have long back noted that the trend of substance use was on the rise particularly in the student population. To find out the prevalence and determinants of smoking practices among undergraduate medical students. A cross-sectional study was conducted among undergraduate medical (MBBS) students of a tertiary care medical college using a predesigned and pretested semi-structured self-administered anonymous questionnaire. Among 182 participants, 55 (30%) were smokers; 85.45% were regular smokers; majority in the age group 20-22 years (70%); mostly males (98%). No significant difference was observed among urban and rural students, and religion had no association. The practice of smoking for last 6 months to 1 year was in 43.6% and 40% smoked less than 6 months. Half of them (50.9%) smoked 5-9 cigarettes per day. Peer pressure was significantly high in smokers (83.6%); 42% had other addictions. The effect of parental smoking on smoking habits of the participants was quite evident among smokers (82%), which was significantly higher than nonsmokers (χ(2) = 63.49, P < 0.05). Peer pressure was the most important risk factor (57.69%) of initiation of smoking habit followed by parental influence (16.49%). Among morbidities of smokers, 60.6% were suffering from regular cough, 6% from bronchitis, and 2% had asthma. Our survey conducted on budding doctors surprisingly showed that undergraduate medical students smoke so much.


The Escherichia coli DinD protein modulates RecA activity by inhibiting postsynaptic RecA filaments.
Escherichia coli dinD is an SOS gene up-regulated in response to DNA damage. We find that the purified DinD protein is a novel inhibitor of RecA-mediated DNA strand exchange activities. Most modulators of RecA protein activity act by controlling the amount of RecA protein bound to single-stranded DNA by affecting either the loading of RecA protein onto DNA or the disassembly of RecA nucleoprotein filaments bound to single-stranded DNA. The DinD protein, however, acts postsynaptically to inhibit RecA during an on-going DNA strand exchange, likely through the disassembly of RecA filaments. DinD protein does not affect RecA single-stranded DNA filaments but efficiently disassembles RecA when bound to two or more DNA strands, effectively halting RecA-mediated branch migration. By utilizing a nonspecific duplex DNA-binding protein, YebG, we show that the DinD effect is not simply due to duplex DNA sequestration. We present a model suggesting that the negative effects of DinD protein are targeted to a specific conformational state of the RecA protein and discuss the potential role of DinD protein in the regulation of recombinational DNA repair.


Optimizing dyes for dye-sensitized solar cells.
Dye-sensitized solar cells (DSSCs) have emerged as an important cheap photovoltaic technology. Charge separation is initiated at the dye, bound at the interface of an inorganic semiconductor and a hole-transport material. Careful design of the dye can minimize loss mechanisms and improve light harvesting. Mass application of DSSCs is currently limited by manufacturing complexity and long-term stability associated with the liquid redox electrolyte used in the most-efficient cells. In this Minireview, dye design is discussed in the context of novel alternatives to the standard liquid electrolyte. Rapid progress is being made in improving the efficiencies of such solid and quasi-solid DSSCs which promises cheap, efficient, and robust photovoltaic systems.


Society of Behavioral Medicine position statement: retain school meal standards and healthy school lunches.
In 2010, the Healthy, Hunger-Free Kids Act (HHFKA) altered nutrition standards in the U.S. National School Lunch Program in an attempt to promote healthy eating and improve children's overall health. However, it was reported that these nutrition standard changes were leading to lower consumption of meals and an increase in plate waste. Initial research was unable to validate these reports. Despite these unsubstantiated claims, the U.S. Department of Agriculture released a proposal to roll back some of the nutritional standards implemented by the HHFKA in 2017. An exploration of the current literature uncovered several new studies that further support the updated meal standards outlined in the HHFKA. Identifying and reviewing these studies was the aim of the current brief. Several new studies found that the implementation of the HHFKA standards led to healthier meals, increased fruit consumption, lower plate waste, and reductions in sodium and calories from saturated fats, to name a few. Given the continued evidence that the HHFKA standards improve dietary quality of school meals, policymakers, schools, and stakeholders are urged to continue to support the retention of the new meal standards.


Human alpha 1-microglobulin and its relationship to renal function.
To evaluate the clinical usefulness in terms of estimation for glomerular filtration rate (GFR), we determined the levels of alpha 1-microglobulin (alpha 1m) in the serum and urine of patients with various renal diseases and compared with those of beta 2-microglobulin (beta 2m) and creatinine. Serum and urinary alpha 1m levels were measured by using single-radial immunodiffusion method. 24-hour creatinine clearance (Ccr) was used as a indicator of GFR. There was a significant positive correlation between serum alpha 1m and creatinine levels (r = 0.75, p less than 0.001). Serum alpha 1m, beta 2m and creatinine inversely correlated and logarithmically correlated to Ccr as shown in the following equations: log alpha 1m = 2.30 - 0.42 X log Ccr (r = -0.74); log beta 2m = 2.06 - 0.91 X log Ccr (r = -0.92); log creatinine = 1.57 - 0.78 X log Ccr (r = -0.94). Both correlation coefficient and regression coefficient for alpha 1m were rather poor compared to those for beta 2m and creatinine. However, alpha 1m levels started to increase over normal range when Ccr fell below 80 liters/day, while beta 2m and creatinine remained within normal ranges. The daily urinary excretion of alpha 1m was increased in the patients whose Ccr was within normal limits compared to that of healthy control subjects (15.2 +/- 3.2 mg/day, n = 19, vs. 5.7 +/- 0.7, n = 7, p less than 0.001). Fractional clearance of alpha 1m increased proportionally to the decrease of Ccr. These data suggest that combined measurements of alpha 1m in the serum and urine seem to be useful to estimate GFR, especially to detect the mild reduction of CFR.


Full Digital Workflow of Nasoalveolar Molding Treatment in Infants With Cleft Lip and Palate.
The aim of this pilot study was to illustrate the feasibility of a full digital workflow to design and manufacturing a consecutive series of customized nasoalveolar molding (NAM) appliances in advance for presurgical unilateral and bilateral cleft lip and palate (CLP) treatment. The full digital workflow consisted of acquisition of 3D image data of an infant's maxilla by using intraoral scanner (TRIOS; 3Shape, Copenhagen, Denmark); the initial data were imported into an appropriate computer-aided design (CAD) software environment, the digital model was virtual modified to achieve a harmonic alveolar arch, and generated a consecutive of digital models of each movement stage; the digital model of NAM appliance was designed based on the virtual modified model; bio-compatible material MED610 was used to manufacturing the real NAM appliances by 3D printing. A consecutive series of NAM appliances was delivered to CLP infant before lip surgery. Intraoral scanning was harmless and safer than conventional impression technique for CLP infants. The CAD/3D printing procedures allowed a series of NAM appliances to be designed and manufactured in advance. The clinical results showed that this full digital workflow was efficient, viable and able to estimate the treatment objective. By the end of presurgical NAM treatment, the malposition alveolar segments had been aligned normally, the surrounding soft tissues repositioned. The full digital workflow presented has provided the potential for presurgical NAM treatment of infants with cleft lip and palate. Intraoral scanning served as a starting point, so subsequent virtual treatment planning and CAD/3D printing procedures could realize the full digital workflow, a whole series of customized NAM appliances was manufactured in advance. This method brings the benefits of safety, affectivity and time-saving.


The low density lipoprotein receptor-related protein complexes with cell surface heparan sulfate proteoglycans to regulate proteoglycan-mediated lipoprotein catabolism.
It has been proposed that clearance of cholesterol-enriched very low density lipoprotein (VLDL) particles occurs through a multistep process beginning with their initial binding to cell-surface heparan sulfate proteoglycans (HSPG), followed by their uptake into cells by a receptor-mediated process that utilizes members of the low density lipoprotein receptor (LDLR) family, including the low density lipoprotein receptor-related protein (LRP). We have further explored the relationship between HSPG binding of VLDL and its subsequent internalization by focusing on the LRP pathway using a cell line deficient in LDLR. In this study, we show that LRP and HSPG are part of a co-immunoprecipitable complex at the cell surface demonstrating a novel association for these two cell surface receptors. Cell surface binding assays show that this complex can be disrupted by an LRP-specific ligand binding antagonist, which in turn leads to increased VLDL binding and degradation. The increase in VLDL binding results from an increase in the availability of HSPG sites as treatment with heparinase or competitors of glycosaminoglycan chain addition eliminated the augmented binding. From these results we propose a model whereby LRP regulates the availability of VLDL binding sites at the cell surface by complexing with HSPG. Once HSPG dissociates from LRP, it is then able to bind and internalize VLDL independent of LRP endocytic activity. We conclude that HSPG and LRP together participate in VLDL clearance by means of a synergistic relationship.


April 2002: 35-year-old healthy man with enlarging right parotid mass.
The April 2002 Case of the Month (COM). 35-year-old healthy man developed a mass in the right parotid gland. A superifical parotidectomy was performed for a 4.5 x 1.5 x 1.5 cm mass involving the intraparotid facial nerve. Grossly the tumor was multinodular, smooth and yellow with normal surrounding salivary gland. Microscopically, the tumor showed expanding nodules composed of proliferating fibroblasts, Schwann cells, and perineural-like cells in a myxoid stroma. Normal peripheral nerve twigs were identified in the periphery of the tumor. There was no increased mitotic activity, cellularity or nuclear pleomorphism. S-100 immunohistochemical stain was positive. The tumor was diagnosed as a solitary plexiform neurofibroma. Plexiform neurofibromas in this area have been described in children with von Recklinghausen's disease or neurofibromatosis 1 (NF 1). Plexiform neurofibromas typically involve deep seated nerve trunks and is considered pathognomonic for NF 1. This unusual case represents a solitary variant of plexiform neurofibroma presenting as a parotid mass in an adult patient without a personal stigmata or family history of NF 1.


A single amino acid controls the functional switch of human constitutive androstane receptor (CAR) 1 to the xenobiotic-sensitive splicing variant CAR3.
The constitutive androstane receptor (CAR) is constitutively activated in immortalized cell lines independent of xenobiotic stimuli. This feature of CAR has limited its use as a sensor for xenobiotic-induced expression of drug-metabolizing enzymes. Recent reports, however, reveal that a splicing variant of human CAR (hCAR3), which contains an insertion of five amino acids (APYLT), exhibits low basal but xenobiotic-inducible activities in cell-based reporter assays. Nonetheless, the underlying mechanisms of this functional shift are not well understood. We have now generated chimeric constructs containing various residues of the five amino acids of hCAR3 and examined their response to typical hCAR activators. Our results showed that the retention of alanine (hCAR1+A) alone is sufficient to confer the constitutively activated hCAR1 to the xenobiotic-sensitive hCAR3. It is noteworthy that hCAR1+A was significantly activated by a series of known hCAR activators, and displayed activation superior to that of hCAR3. Moreover, intracellular localization assays revealed that hCAR1+A exhibits nuclear accumulation upon 6-(4-chlorophenyl) imidazo[2,1-b][1,3]thiazole-5-carbaldehyde-O-(3,4-dichlorobenzyl) oxime (CITCO) treatment in COS1 cells, which differs from the spontaneous nuclear distribution of hCAR1 and the nontranslocatable hCAR3. Mammalian two-hybrid and glutathione S-transferase pull-down assays further demonstrated that hCAR1+A interacts with the coactivator SRC-1 and GRIP-1 at low level before activation, while at significantly enhanced level in the presence of CITCO. Thus, the alanine residue in the insertion of hCAR3 seems in charge of the xenobiotic response of hCAR3 through direct and indirect mechanisms. Activation of hCAR1+A may represent a sensitive avenue for the identification of hCAR activators.


Mining the Giardia genome and proteome for conserved and unique basal body proteins.
Giardia lamblia is a flagellated protozoan parasite and a major cause of diarrhoea in humans. Its microtubular cytoskeleton mediates trophozoite motility, attachment and cytokinesis, and is characterised by an attachment disk and eight flagella that are each nucleated in a basal body. To date, only 10 giardial basal body proteins have been identified, including universal signalling proteins that are important for regulating mitosis or differentiation. In this study, we have exploited bioinformatics and proteomic approaches to identify new Giardia basal body proteins and confocal microscopy to confirm their localisation in interphase trophozoites. This approach identified 75 homologs of conserved basal body proteins in the genome including 65 not previously known to be associated with Giardia basal bodies. Thirteen proteins were confirmed to co-localise with centrin to the Giardia basal bodies. We also demonstrate that most basal body proteins localise to additional cytoskeletal structures in interphase trophozoites. This might help to explain the roles of the four pairs of flagella and Giardia-specific organelles in motility and differentiation. A deeper understanding of the composition of the Giardia basal bodies will contribute insights into the complex signalling pathways that regulate its unique cytoskeleton and the biological divergence of these conserved organelles.


Photothermally induced Bergman cyclization of metalloenediynes via near-infrared ligand-to-metal charge-transfer excitation.
Reaction of 1,2-bis(tert-butyldimethylsilyloxy)-4,5-diiodobenzene with 2 equiv of phenylacetylene followed by deprotection with KF/HBr yields the catechol-enediyne ligand 4,5-bis(phenylethynyl)benzene-1,2-diol (CatED, 1). Metathesis of VO(SALIMH)ACAC.CH(3)OH (2) with 1 and subsequent air oxidation yields (4,5-bis(phenylethynyl)-1,2-dihydroxyphenyl)[4-(2-(salicylideneamino)ethyl)imidazolyl]oxovanadium(V).CH(3)OH [VO(SALIMH)CatED], (3), in 85%. The thermal Bergman cyclization temperature for 3 is very high (246 degrees C), which is expected for a rigid, benzannulated enediyne motif. The electronic spectrum of 3 exhibits two strong ligand-to-metal charge transfer (LMCT) transitions centered at 584 nm (epsilon = 6063 M(-)(1) cm(-)(1)) and 1028 nm (epsilon = 8098 M(-)(1) cm(-)(1)). These transitions derive from CatED-to-V(V) ligand-to-metal charge transfer, the assignment of which is verified by resonance enhancement of several CatED vibrational modes in the Raman spectra obtained with lambda = 785 vs lambda = 457.9 nm under low power and/or temperature conditions. At elevated temperatures (113-323 K) and powers (2-5 mW), excitation of 3 in the solid state with lambda = 785 nm leads to generation of a black, sparingly soluble, fluorescent product that exhibits weak vibrational features in the 580-600, 1200-1350, and 1450-1600 cm(-)(1) regions, indicative of V-O (CatED) and aromatic ring units. The C=C ring modes correspond well with the vibrational characteristics of poly(p-phenylene) and derivatives thereof. Additionally, materials generated in both the solid-state thermal and photothermal reactions of 3 demonstrate the formation of high molecular weight species ranging from 5000 to 274 000. On the basis of these data and the literature precedent for formation of poly(p-phenylene) via thermolysis of simple enediynes, the reaction poses a unique approach for photoinitiating Bergman cyclization with long-wavelength excitation, as well as the generation of polymeric products.


Escherichia coli YtfE is a di-iron protein with an important function in assembly of iron-sulphur clusters.
Our previous analysis of the transcriptome of Escherichia coli under nitrosative stress showed that the ytfE gene was one of the highest induced genes. Furthermore, the E. coli strain mutated on the ytfE gene was found to be more sensitive to nitric oxide than the wild-type strain. In the present work, we show that the mutation of the ytfE gene in E. coli yielded a strain that grows poorly under anaerobic respiratory conditions and that has an increased sensitivity to iron starvation. Furthermore, all examined iron-sulphur proteins have decreased activity levels in the strain lacking ytfE. Altogether, the results suggest a role for ytfE in iron-sulphur cluster biogenesis. YtfE was overexpressed in E. coli and it is shown to contain a di-iron centre of the histidine-carboxylate family.


Homology between rat liver RNA populations during development, regeneration, and neoplasia.
To investigate the degree of homology which may exist between rat liver RNA populations during development, regeneration, and neoplasia, we hybridized polyadenylated RNAs from (a) normal adult, (b) 24-h regenerating, (c) 20-day fetal livers, and (d) the transplantable Morris hepatoma 5123tc to homologous and heterologous complementary DNAs and to cDNAs enriched for sequences preferentially transcribed in either adult or fetal liver. We also compared the in vitro translation products of these RNAs. Analyses of normal adult, regenerating, and fetal liver RNA populations and their translation products show that the overall pattern of gene expression during liver regeneration differs little from that of normal adult rat liver and that mature hepatocytes do not appear to revert to an "immature" state upon reentering the cell cycle. Comparisons between fetal, normal adult, and tumor RNA populations revealed that RNA populations from fetal liver and the 5123tc tumor lack sequences normally expressed in the mature adult liver. However, the tumor does not "reexpress" sequences which are preferentially expressed in fetal livers.


Further Analyses of Genetic Association Between GRM8 and Alcohol Dependence Symptoms Among Young Adults.
The gene GRM8, a metabotropic glutamate receptor, has emerged as a gene of interest for its possible role in the development of alcohol dependence, with evidence of association with an electrophysiological endophenotype and level of response to alcohol as well as suggestive evidence of association with alcohol dependence. The present study further investigated the association between GRM8 and alcohol dependence symptom counts among young adults using a new sample of individuals collected as part of the prospective sample (ages 18-26 years; N = 842) from the Collaborative Study on the Genetics of Alcoholism (COGA). Two single-nucleotide polymorphisms were significantly associated with alcohol dependence in European Americans using the Nyholt corrected p value of .007: rs886003 (β = -.212, p = .0002) and rs17862325 (β = -.234, p < .0001), but not in African Americans, likely because of the lower power to detect association in this group. These results further implicate the role of glutamate receptor genes such as GRM8 in the development of alcohol dependence.


Effects of ischemia on sarcoplasmic reticulum Ca(2+) uptake and Ca(2+) release in rat skeletal muscle.
In this study, we investigated the hypothesis that prolonged ischemia would impair both sarcoplasmic reticulum (SR) Ca(2+) uptake and Ca(2+) release in skeletal muscle. To induce total ischemia (I), a tourniquet was placed around the upper hindlimb in 30 female Sprague-Dawley rats [wt = 256 +/- 6.7 (SE) g] and inflated to 350 mmHg for 4 h. The contralateral limb served as control (C). Immediately after the 4 h of ischemia, mixed gastrocnemius and tibialis anterior muscle was sampled from both limbs, and both crude muscle homogenates and SR vesicles were prepared. In another 10 control animals (CC), muscles were sampled and prepared exactly the same way, but immediately after anesthetization. Ca(2+) uptake and Ca(2+) release were measured in vitro with Indo-I on both homogenates and SR vesicles. As hypothesized, submaximal Ca(2+) uptake was lower (P < 0.05) in I compared with CC and C, by 25 and 45% in homogenates and SR vesicles, respectively. Silver nitrate (AgNO(3))-induced Ca(2+) release, which occurred in two phases (phase 1 and phase 2), was also altered in I compared with CC and C, in both muscle homogenates and SR vesicles. With ischemia, phase 1 peak Ca(2+) release was 26% lower (P < 0.05) in SR vesicles only. For phase 2, peak Ca(2+) release was 54 and 24% lower (P < 0.05) in SR vesicles and homogenates, respectively. These results demonstrate that prolonged skeletal muscle ischemia leads to a reduced SR Ca(2+) uptake in both homogenates and SR vesicles. The effects of ischemia on SR Ca(2+) release, however, depend on both the phase examined and the type of tissue preparation.


Rac1 is required for cardiomyocyte apoptosis during hyperglycemia.
Hyperglycemia induces reactive oxygen species (ROS) and apoptosis in cardiomyocytes, which contributes to diabetic cardiomyopathy. The present study was to investigate the role of Rac1 in ROS production and cardiomyocyte apoptosis during hyperglycemia. Mice with cardiomyocyte-specific Rac1 knockout (Rac1-ko) were generated. Hyperglycemia was induced in Rac1-ko mice and their wild-type littermates by injection of streptozotocin (STZ). In cultured adult rat cardiomyocytes, apoptosis was induced by high glucose. The results showed a mouse model of STZ-induced diabetes, 7 days of hyperglycemia-upregulated Rac1 and NADPH oxidase activation, elevated ROS production, and induced apoptosis in the heart. These effects of hyperglycemia were significantly decreased in Rac1-ko mice or wild-type mice treated with apocynin. Interestingly, deficiency of Rac1 or apocynin treatment significantly reduced hyperglycemia-induced mitochondrial ROS production in the heart. Deficiency of Rac1 also attenuated myocardial dysfunction after 2 months of STZ injection. In cultured cardiomyocytes, high glucose upregulated Rac1 and NADPH oxidase activity and induced apoptotic cell death, which were blocked by overexpression of a dominant negative mutant of Rac1, knockdown of gp91(phox) or p47(phox), or NADPH oxidase inhibitor. In type 2 diabetic db/db mice, administration of Rac1 inhibitor, NSC23766, significantly inhibited NADPH oxidase activity and apoptosis and slightly improved myocardial function. Rac1 is pivotal in hyperglycemia-induced apoptosis in cardiomyocytes. The role of Rac1 is mediated through NADPH oxidase activation and associated with mitochondrial ROS generation. Our study suggests that Rac1 may serve as a potential therapeutic target for cardiac complications of diabetes.