3-aminobenzamide, one of poly(ADP-ribose)polymerase-1 inhibitors, rescuesapoptosisin rat models of spinal cord injury.
Poly(ADP-ribose)polymerase-1 (PARP-1) is anubiquitous, DNA repair-associated enzyme, which participates in gene expression, cell death, central nerve system (CNS) disorders and oxidative stress. According to the previous studies, PARP-1 over-activation may lead to over-consumption of ATP and even cell apoptosis. Spinal cord injury (SCI) is an inducement towards PARP-1 over-activation due to its massive damage to DNA. 3-aminobenzamide (3-AB) is a kind of PARP-1 inhibitors. The relationship among PARP-1, 3-AB, SCI and apoptosis has not been fully understood. Hence, in the present study, we focused on the effects of 3-AB on cell apoptosis after SCI. Accordingly, SCI model was constructed artificially, and 3-AB was injected intrathecally into the Sprague-Dawley (SD) rats. The results demonstrated an increase in cell apoptosis after SCI. Furthermore, PARP-1 was over-activated after SCI but inhibited by 3-AB injection. In addition, apoptosis-inducing factor (AIF) was inhibited but B-cell lymphoma-2 (Bcl-2) was up-regulated by 3-AB. Interestingly, caspase-3 was not significantly altered with or without 3-AB. In conclusion, our experiments showed that 3-AB, as a PARP-1 inhibitor, could inhibit cell apoptosis after SCI in caspase-independent way, which could provide a better therapeutic target for the treatment of SCI.


Reverse cholesterol transport: from classical view to new insights.
Cholesterol is of vital importance for the human body. It is a constituent for most biological membranes, it is needed for the formation of bile salts, and it is the precursor for steroid hormones and vitamin D. However, the presence of excess cholesterol in cells, and in particular in macrophages in the arterial vessel wall, might be harmful. The accumulation of cholesterol in arteries can lead to atherosclerosis, and in turn, to other cardiovascular diseases. The route that is primarily thought to be responsible for the disposal of cholesterol is called reverse cholesterol transport (RCT). Therefore, RCT is seen as an interesting target for the development of drugs aimed at the prevention of atherosclerosis. Research on RCT has taken off in recent years. In this review, the classical concepts about RCT are discussed, together with new insights about this topic.


Compartmentalization of algal bioluminescence: autofluorescence of bioluminescent particles in the dinoflagellate Gonyaulax as studied with image-intensified video microscopy and flow cytometry.
Compartmentalization of specialized functions to discrete locales is a fundamental theme of eucaryotic organization in cells. We report here that bioluminescence of the dinoflagellate alga Gonyaulax originates in vivo from discrete subcellular loci that are intrinsically fluorescent. We demonstrate this localization by comparing the loci of fluorescence and bioluminescence as visualized by image-intensified video microscopy. These fluorescent particles appeared to be the same as the previously described in vitro "scintillons." We attribute the endogenous fluorescence to that of the bioluminescence substrate, luciferin, because (a) the fluorescence excitation and emission characteristics are comparable, (b) the autofluorescence is lost after exhaustive stimulation of bioluminescence, and (c) the fluorescence of discharged particles in vitro can be restored by adding luciferin. The fluorescence in vivo exhibits a standard property of circadian (daily) rhythmicity: under constant environmental conditions, the intensity of the particle fluorescence fluctuates cyclically (it is maximal during the night phase and is low during the day). Thus, luciferin is localized within the cell at discrete loci from which the bioluminescence emanates; the cellular quantity of luciferin is rhythmically modulated by the circadian clock.


Achievements of the COVID-19 Turkey Platform in vaccine and drug development with an approach of "co-creation and succeeding together".
Mobilizing the research ecosystem for accelerating vaccine and drug development has been an important reality of the pandemic. This article reviews the scientific advances that are attained by the COVID-19 Turkey Platform for vaccine and drug development against the SARS-CoV-2 virus. The platform that is coordinated by the Scientific and Technological Research Council of Turkey is established with a "co-creation and succeeding together" approach, which involves 436 researchers across 49 different institutions working on 17 vaccine and drug development projects in total. Recent advances of the COVID-19 Turkey Platform include the fourth virus-like particle-based vaccine candidate in the world to enter clinical studies based on the World Health Organization COVID-19 vaccine tracker that is currently completing phase 2 clinical studies on the path towards initiating phase 3 clinical studies. Moreover, an adjuvanted inactivated vaccine candidate and two drug candidates that have been identified through the virtual scanning of more than 20,000 molecules are currently in clinical studies. Other vaccines and drug candidates involve additional innovative aspects, and a locally synthesized drug is found to have an impact on COVID-19. This review article discusses the advances that are achieved by the COVID-19 Turkey Platform from the ecosystem perspective, emphasizing the important scientific advances that have been achieved in the field of medical sciences.


Intravesical migration of a GyneFix intrauterine device.
A case of intravesical migration of a GyneFix intrauterine device (IUD) is described, in which the patient presented with supra-pubic pain and urinary symptoms. The diagnosis was made 34 months after the insertion of the IUD, by ultrasound scan. The GyneFix was removed endoscopically. A description of the GyneFix device, the possible adverse effects and incidences of its complications, the importance of post-insertion follow-up, and the need for awareness of the possibility of intravesical migration are discussed.


Synthesis of 3-substituted 3-fluoro-2-oxindoles by deacylative alkylation.
The fluorination of 3-acetyl-2-oxindoles with N-fluorobenzenesulfonimide under Lewis acid catalysis using Mg(ClO4)2 gives the starting compounds 3-acetyl-3-fluoro-2-oxindoles. These compounds are subjected to base-promoted deacylative alkylation (DaA) for the in situ generation of 3-fluoro-2-oxindole enolates under very mild reaction conditions using Triton B (1 equiv.) and alkyl halides and Michael acceptors as electrophilic reagents. The corresponding 3-alkylated-3-fluoro-2-oxindoles are obtained in good to very high yields. In addition, the palladium-catalyzed deacylative allylation is carried out with allylic alcohols using LiOtBu as the base and 6 mol% of Pd(OAc)2 and dppp, giving the resulting 3-allylated 3-fluoro-2-oxindoles in good yields. This methodology allows a simple synthesis of 3-alkylated-3-fluoro-2-oxindoles, which are difficult to obtain by other routes.


Selective Nonmethylated CpG DNA Recognition Mechanism of Cysteine Clamp Domains.
Methylation of DNA at CpG sites is a major mark for epigenetic regulation, but how transcription factors are influenced by CpG methylation is not well understood. Here, we report the molecular mechanisms of how the TCF (T-cell factor) and GEF (glucose transporter 4 enhancer factor) families of proteins selectively target unmethylated DNA sequences with a C-clamp type zinc finger domain. The structure of the C-clamp domain from human GEF family protein HDBP1 (C-clampHDBP1) in complex with DNA was determined using NMR spectroscopy, which adopts a unique zinc finger fold and selectively binds RCCGG (R = A/G) DNA sequences with an "Arg···Trp-Lys-Lys" DNA recognition motif inserted in the major groove. The CpG base pairs are central to the binding due to multiple hydrogen bonds formed with the backbone carbonyl groups of Trp378 and Lys379, as well as the side chain ε-amino groups of Lys379 and Lys380 from C-clampHDBP1. Consequently, methylation of the CpG dinucleotide almost abolishes the binding. Homology modeling reveals that the C-clamp domain from human TCF1E (C-clampTCF1E) binds DNA through essentially the same mechanism, with a similar "Arg···Arg-Lys-Lys" DNA recognition motif. The substitution of tryptophan by arginine makes C-clampHDBP1 prefer RCCGC DNA sequences. The two signature DNA recognition motifs are invariant in the GEF and TCF families of proteins, respectively, from fly to human. The recognition of the CpG dinucleotide through two consecutive backbone carbonyl groups is the same as that of the CXXC type unmethylated CpG DNA binding domains, suggesting a common mechanism shared by unmethylated CpG binding proteins.


Composition of the mount st. Helens ashfall in the moscow-pullman area on 18 may 1980.
Mineralogical and chemical analyses of the ashfall from Mount St. Helens on 18 May 1980 indicate that there were two distinct ashes. The early dark ash is composed principally of plagioclase and lithic fragments of plagioclase and glass with titanium-rich magnetite and some basaltic hornblende and orthopyroxene. The later pale ash, four-fifths by weight of the whole fallout, is 80 percent glass with plagioclase as the principal crystalline phase. Quartz and potassium feldspar are rare to absent in both ashes. Chemical analyses of nine ash fractions and of the glass in each type emphasize the differences between the two ash types and their chemical homogeneity.


Plant-based production and characterization of a promising Fc-fusion protein against microgravity-induced bone density loss.
Microgravity-induced bone loss is a main obstacle for long term space missions as it is difficult to maintain bone mass when loading stimuli is reduced. With a typical bone mineral density loss of 1.5% per month of microgravity exposure, the chances for osteoporosis and fractures may endanger astronauts' health. Parathyroid Hormone or PTH (1-34) is an FDA approved treatment for osteoporosis, and may reverse microgravity-induced bone loss. However, PTH proteins requires refrigeration, daily subcutaneous injection, and have a short shelf-life, limiting its use in a resource-limited environment, like space. In this study, PTH was produced in an Fc-fusion form via transient expression in plants, to improve the circulatory half-life which reduces dosing frequency and to simplify purification if needed. Plant-based expression is well-suited for space medicine application given its low resource consumption and short expression timeline. The PTH-Fc accumulation profile in plant was established with a peak expression on day 5 post infiltration of 373 ± 59 mg/kg leaf fresh weight. Once the PTH-Fc was purified, the amino acid sequence and the binding affinity to its target, PTH 1 receptor (PTH1R), was determined utilizing biolayer interferometry (BLI). The binding affinity between PTH-Fc and PTH1R was 2.30 × 10-6 M, similar to the affinity between PTH (1-34) and PTH1R (2.31 × 10-6 M). Its function was also confirmed in a cell-based receptor stimulation assay, where PTH-Fc was able to stimulate the PTH1R producing cyclic adenosine monophosphate (cAMP) with an EC50 of (8.54 ± 0.12) x 10-9 M, comparable to the EC50 from the PTH (1-34) of 1.49 × 10-8 M. These results suggest that plant recombinant PTH-Fc exhibits a similar binding affinity and potency in a PTH1R activation assay compared to PTH. Furthermore, it can be produced rapidly at high levels with minimal resources and reagents, making it ideal for production in low resource environments such as space.


Countergradient variation and secondary sexual color: phenotypic convergence promotes genetic divergence in carotenoid use between sympatric anadromous and nonanadromous morphs of sockeye salmon (Oncorhynchus nerka).
Genetically distinct anadromous (sockeye) and nonanadromous (kokanee) morphs of the Pacific salmon, Oncorhynchus nerka, develop identical, brilliant red color at maturity during sympatric breeding in freshwater streams. The marine and lacustrine environments they occupy prior to maturity, however, appear to differ in the availability of dietary carotenoid pigments necessary to produce red coloration. We tested the hypothesis that kokanee, which occupy carotenoid-poor lakes, are more efficient at using the dietary pigments than are sockeye, which occupy the more productive North Pacific Ocean. In a 2-year controlled breeding study, flesh and skin color of mature and immature crosses fed a low-carotenoid diet were quantified with both a chromameter and by chemical extraction of carotenoid pigments. Results revealed striking countergradient variation in carotenoid use, with kokanee approximately three times more efficient at sequestering the pigments to the flesh musculature than similar age sockeye. This difference translated into virtually nonoverlapping differences between pure crosses in secondary sexual color at maturity, when the pigments are mobilized and transported to the skin. Kokanee crosses turned pinkish red over most of their body, whereas sockeye turned olive green. The olive green was similar to the breeding color of residuals in the wild, the progeny of anadromous sockeye that remain in fresh water and are believed to have given rise to kokanee on numerous independent occasions. Reciprocal hybrids were similar to each other and intermediate to the pure crosses, indicating additive genetic inheritance. Mate choice trials with sockeye males in the wild showed the ancestral morph strongly preferred red over green models. These results suggest a preference for red mates maintained in nonanadromous breeding populations drove the reevolution of the red phenotype in kokanee via more efficient use of dietary carotenoid pigments. This is a novel, yet hidden, mechanism by which sexual selection promotes the genetic differentiation of these sympatric populations.


CXXC5 expression in prostate cancer: implications for cancer progression.
Identification of genes specifically deregulated in prostate adenocarcinoma may lead to discovery of new oncogenes/tumour suppressors with clinical relevance for diagnosis, prognosis and/or therapy. CXXC5 is a gene encoding a retinoid-inducible nuclear factor, whose overexpression in breast tumours, metastatic malignant melanomas and papillary thyroid carcinoma has been recently reported. We previously found differential expression of CXXC5 transcripts in metastatic prostate cancer cell lines of both rat and human origin. However, knowledge on the expression of this gene in benign or malignant human prostate tissue is lacking. The aim of this study was to determine the mRNA and protein expression pattern of CXXC5 in human benign prostate tissue, proliferative inflammatory atrophy, high-grade prostatic intra-epithelial neoplasia and prostate cancer, using qPCR, chromogenic in situ hybridization and immunohistochemistry. Our results showed that protein levels determined by immunohistochemistry were in agreement with transcript levels observed by chromogenic in situ hybridization. CXXC5 mRNA and protein expressions were significantly higher in prostate cancer, high-grade prostatic intra-epithelial neoplasia, and proliferative inflammatory atrophy, compared to benign prostate tissue. Significantly, within the same tissue specimens, CXXC5 staining was stronger in malignant acini than in matched adjacent, benign acini; immunostaining for this protein was mainly localized to the nucleus of benign epithelial cells and both the nucleus and cytoplasm of malignant epithelial cells. Our findings suggest that CXXC5 may play a role in the process of prostate carcinogenesis. Additional studies are required to determine the biological and clinical significance of CXXC5 in prostate cancer development and/or progression.


Trajectories of Self-Reported Opioid Use Among Patients With HIV Engaged in Care: Results From a National Cohort Study.
No prior studies have characterized long-term patterns of opioid use regardless of source or reason for use among patients with HIV (PWH). We sought to identify trajectories of self-reported opioid use and their correlates among a national sample of PWH engaged in care. Veterans Aging Cohort Study, a prospective cohort including PWH receiving care at 8 US Veterans Health Administration (VA) sites. Between 2002 and 2018, we assessed past year opioid use frequency based on self-reported "prescription painkillers" and/or heroin use at baseline and follow-up. We used group-based trajectory models to identify opioid use trajectories and multinomial logistic regression to determine baseline factors independently associated with escalating opioid use compared to stable, infrequent use. Among 3702 PWH, we identified 4 opioid use trajectories: (1) no lifetime use (25%); (2) stable, infrequent use (58%); (3) escalating use (7%); and (4) de-escalating use (11%). In bivariate analysis, anxiety; pain interference; prescribed opioids, benzodiazepines and gabapentinoids; and marijuana use were associated with escalating opioid group membership compared to stable, infrequent use. In multivariable analysis, illness severity, pain interference, receipt of prescribed benzodiazepine medications, and marijuana use were associated with escalating opioid group membership compared to stable, infrequent use. Among PWH engaged in VA care, 1 in 15 reported escalating opioid use. Future research is needed to understand the impact of psychoactive medications and marijuana use on opioid use and whether enhanced uptake of evidence-based treatment of pain and psychiatric symptoms can prevent escalating use among PWH.


Oral fluid replacement by a glucose glycine electrolyte formulation in E coli and rotavirus diarrhoea in pigs.
A glucose glycine electrolyte solution (GGES) was investigated to determine its suitability for oral rehydration in diarrhoea in pigs. Piglets with diarrhoea following experimental infection with eneteropathogenic Escherichia coli were given access in cube drinkers to either GGES or water. The mortality in the GGES group (11.6 per cent) was significantly (P less than 0.05) less than in the controls (24.0 per cent), and weight gain in severely diarrhoeic piglets was greater in the GGES group. Gnotobiotic piglets were challenged with pig rotavirus and either given access to GGES or milk. The GGES was either in restricted volume or ad lib. Restricted access to GGES prevented much of the weight loss due to the diarrhoea and ad lib access to GGES allowed relatively uninterrupted weight gain in comparison with controls. In naturally occurring cases of diarrhoea on farms, access to GGES reduced mortality to weaning from 19.7 per cent to 7.2 per cent (P less than 0.05). Weight gain per day was also greater in piglets which scoured for three days or more, indicating the rehydrating effect in reducing net fluid loss. Thus GGES treatment was of value in treatment e&ects of diarrhoea due to either E. coli or rotavirus.


Targetoid form of polymorphic eruption of pregnancy: A case report.
Polymorphic eruption of pregnancy, formerly known as pruritic and urticarial papules and plaques of pregnancy, is an uncommon cutaneous eruption that can affect women during their third trimester of pregnancy. As its name implies, it has a variety of morphologies and it is important to consider other diagnoses, such as pemphigoid gestationis, which polymorphic eruption of pregnancy can mimic. Sometimes, as in this case, polymorphic eruption of pregnancy can have a targetoid morphology reminiscent of erythema multiforme. A thorough workup and conservative management plan helps reassure the patient that the correct approach is being taken during the challenging period of a pregnancy nearing completion.


Improving the accuracy of template-based predictions by mixing and matching between initial models.
Comparative modeling is a technique to predict the three dimensional structure of a given protein sequence based primarily on its alignment to one or more proteins with experimentally determined structures. A major bottleneck of current comparative modeling methods is the lack of methods to accurately refine a starting initial model so that it approaches the resolution of the corresponding experimental structure. We investigate the effectiveness of a graph-theoretic clique finding approach to solve this problem. Our method takes into account the information presented in multiple templates/alignments at the three-dimensional level by mixing and matching regions between different initial comparative models. This method enables us to obtain an optimized conformation ensemble representing the best combination of secondary structures, resulting in the refined models of higher quality. In addition, the process of mixing and matching accumulates near-native conformations, resulting in discriminating the native-like conformation in a more effective manner. In the seventh Critical Assessment of Structure Prediction (CASP7) experiment, the refined models produced are more accurate than the starting initial models. This novel approach can be applied without any manual intervention to improve the quality of comparative predictions where multiple template/alignment combinations are available for modeling, producing conformational models of higher quality than the starting initial predictions.


Keratinocyte growth factor protects murine hepatocytes from tumor necrosis factor-induced apoptosis in vivo and in vitro.
Keratinocyte growth factor (KGF) promotes epithelial growth and differentiation and has potent effects on the liver. The coinjection of lipopolysaccharide (LPS) and D-galactosamine (GalN) results in hepatic failure in mice. Mechanistically, LPS-induced tumor necrosis factor (TNF) triggers hepatocyte apoptosis, which is enhanced by GalN-arrested transcription. Similarly, the combination of TNF and actinomycin D (ActD) causes hepatocyte apoptosis in vitro. We studied the effect of KGF on LPS and GalN-induced hepatic failure in vivo and on TNF- and ActD-induced hepatocyte apoptosis in vitro, where it was compared with those of hepatic growth factor (HGF) and epidermal growth factor (EGF). Mice treated with human recombinant KGF (1 mg/kg subcutaneously) 24 hours before intraperitoneal coinjection of LPS and GalN sustained prolonged survival compared with control mice, although overall mortality was not changed. The counts of apoptotic hepatocytes, serum alanine and aspartate transaminases, and DNA fragments in the cytosolic fraction of liver homogenates were higher in control mice than in treated mice 6 hours after LPS and GalN coinjection, before any mortality occurred. In vitro, hepatocytes pretreated with KGF exhibited reduced TNF- and ActD-induced cell damage and DNA fragmentation, similar to hepatocytes pretreated with HGF and EGF. In conclusion, KGF prolongs survival during LPS- and GalN-induced hepatic failure by temporarily protecting hepatocytes against apoptosis. It also protects hepatocytes in vitro against TNF- and ActD-induced apoptosis.


Long-term Adjustment After Surviving Open Heart Surgery: The Effect of Using Prayer for Coping Replicated in a Prospective Design.
despite the growing evidence for effects of religious factors on cardiac health in general populations, findings are not always consistent in sicker and older populations. We previously demonstrated that short-term negative outcomes (depression and anxiety) among older adults following open heart surgery are partially alleviated when patients employ prayer as part of their coping strategy. The present study examines multifaceted effects of religious factors on long-term postoperative adjustment, extending our previous findings concerning prayer and coping with cardiac disease. analyses capitalized on a preoperative survey and medical variables from the Society of Thoracic Surgeons' National Database of patients undergoing open heart surgery. The current participants completed a mailed survey 30 months after surgery. Two hierarchical regressions were performed to evaluate the extent to which religious factors predicted depression and anxiety, after controlling for key demographics, medical indices, and mental health. predicting lower levels of depression at the follow-up were preoperative use of prayer for coping, optimism, and hope. Predicting lower levels of anxiety at the follow-up were subjective religiousness, marital status, and hope. Predicting poorer adjustment were reverence in religious contexts, preoperative mental health symptoms, and medical comorbidity. Including optimism and hope in the model did not eliminate effects of religious factors. Several other religious factors had no long-term influences. MPLICATIONS: the influence of religious factors on the long-term postoperative adjustment is independent and complex, with mediating factors yet to be determined. Future research should investigate mechanisms underlying religion-health relations.


Influence of growth rate on nitrogen balance in adolescent sprint athletes.
This study aimed to estimate nitrogen balance and protein requirements in adolescent sprint athletes as a function of growth rate and physical development. Sixty adolescent sprint athletes were followed up biannually over a 2-yr period. Individual growth curves and age at peak height velocity were determined. Skeletal muscle mass (SMM) was estimated based on anthropometric measurements and fat mass was estimated by underwater densitometry. Seven-day diet and physical activity diaries were completed to estimate energy balance and protein intake. Nitrogen analysis of 24-hr urine samples collected on 1 weekday and 1 weekend day allowed calculation of nitrogen balance. Body height, weight, and SMM increased throughout the study period in both genders. Mean protein intakes were between 1.4 and 1.6 g kg-1 day-1 in both genders. A protein intake of 1.46 g kg-1 day-1 in girls and 1.35 g kg-1 day-1 in boys was needed to yield a positive nitrogen balance. This did not differ between participants during and after their growth spurt. None of the growth parameters was significantly related to nitrogen balance. It can be concluded that a mean protein intake around 1.5 g kg-1 day-1 was sufficient to stay in a positive nitrogen balance, even during periods of peak growth. Therefore, protein intake should not be enhanced in peak periods of linear or muscular growth.


Streamlining stroke care: from symptom onset to emergency department.
Prehospital delays in the treatment of stroke patients, including identification of stroke as a medical emergency, represent a significant and preventable obstacle to optimal stroke care. Although patient delay in seeking care represents the greatest barrier to expedient care, delays often exist in the identification, transport, and triage of stroke patients. Public education in recognizing stroke symptoms as warranting immediate care and appropriate training of emergency medical service personnel are essential parts of community-wide, coordinated stroke care. In addition, emergency physicians must be engaged in the effort to limit delays if the rates of patients eligible for thrombolytic therapy are to improve. This review presents the common inadequacies in the prehospital identification and care for stroke patients and discusses changes within the community health care system that can be implemented to improve the critical early stages of stroke management.


Effects of knee surgery on cardiac function in soccer players.
To evaluate the effects of knee surgery on hematocrit and hemoglobin concentration and on resting cardiac parameters as measured by echocardiography. Ten soccer players who underwent knee surgery were evaluated before (T1) and after (T2) hospitalization within a 7-day interval. After hospitalization, end diastolic volume and stroke volume were significantly reduced (P < 0.05) by 14 and 22%, respectively. Despite a significant increase in resting heart rate (T1: 68 +/- 3.3 beats/ min, T2: 72 +/- 3.1 beats/min, P < 0.05), cardiac output was significantly decreased (T1: 4.89 +/- 0.56 liters/min; 3.95 +/- 0.62 liters/min, P < 0.05). The ejection fraction was 65% at T1 and fell to 58% at T2 (P < 0.05). After hospitalization, significant decreases in hemoglobin concentration and hematocrit were observed, suggesting a fall in blood volume. In soccer players, knee surgery leads to resting cardiac deconditioning, which is characterized by a significant reduction in stroke volume.


Decitabine represses osteoclastogenesis through inhibition of RANK and NF-κB.
DNA methylation is essential for maintenance of stable repression of gene transcription during differentiation and tumorigenesis. Demethylating reagents including decitabine could release the repression, leading to perturbed transcription program. Recently others and we showed that, in B cell lymphomas, decitabine repressed B cell specific gene transcription and activated NF-κB signaling, causing decreased expression of translocated oncogenes including MYC and attenuated tumor cell proliferation. During osteoclastogenesis, changes in DNA methylation occurred in numerous genes, implicating important roles for DNA methylation in osteoclastogenesis. In the present study, we found that decitabine inhibited osteoclastogenesis. The inhibitory effect could be at least partially attributed to reduced expression of multiple osteoclast specific genes including RANK by decitabine. Moreover, decitabine inhibited activity of NF-κB, AP-1 and extracellular signal-regulated kinase (ERK), but not PI3K/Akt pathway. In vivo, using ovariectomized mouse as a model, we observed that decitabine reduced the osteoclast activity and bone loss. In conclusion, our findings demonstrated that decitabine was an inhibitor of osteoclastogenesis by repression of osteoclast specific transcription program including the RANK, NF-κB and AP-1 pathways. DNA methylation might be indispensable for osteoclastogenesis. The use of decitabine could represent a novel strategy in treatment of diseases associated with increased osteoclast activity.


Epidemiology of diarrhea in travelers.
To investigate diarrhea in tourists on a worldwide scale, 16,568 passengers were interviewed during their flights home from 13 destinations in various climatic regions. Significant differences in diarrheal incidence were observed between individual destinations and also between hotels in the same area. The highest rates exceeded 50%. Younger persons were more often affected. Sex, travel characteristics, and a record of former tropical journeys influenced the incidence to a minor degree. Even in the tropics, diarrhea usually takes a short (average, 3.6 days) and mild (average of 4.6 stools per day) course. Prophylactic or therapeutic medication only slightly influenced these values. The various regions showed minor differences in chronology and symptomatology. This is consistent with quantitative rather than qualitative geographic variations in causative agents. The traditional nutritional recommendations for prophylaxis seemed to be unrealistic and usually unsuccessful.


Cefadroxil in the horse: pharmacokinetics and in vitro antibacterial activity.
Sodium cefadroxil was administered as a single intravenous dose (25 mg/kg) to six healthy adult mares. Plasma samples were collected over a 24-h period and cefadroxil concentrations were measured by microbiological assay. The pharmacokinetic behavior of the drug was appropriately described in terms of a one-compartment open model. Values for the major pharmacokinetic terms were: extrapolated initial plasma concentration = 59.2 +/- 15.0 micrograms/ml; half-life = 46 +/- 20 min; apparent volume of distribution = 462 +/- 191 ml/kg; and body clearance = 7.0 +/- 0.6 ml/min.kg. In a subsequent study, a suspension of cefadroxil monohydrate was administered intragastrically (25 mg/kg) to the same six horses. Plasma concentrations of the drug peaked at 1-2 h but, in general, absorption was both poor and inconsistent. The data were unsuitable for determination of cefadroxil bioavailability from this oral dosage form. Ninety-nine isolates of eleven bacterial species obtained from clinically ill horses were tested for susceptibility to cefadroxil. All strains of Streptococcus equi, Streptococcus zooepidemicus, coagulase-positive staphylococci, Corynebacterium pseudotuberculosis and five out of six strains of Actinobacillus suis were highly susceptible to the drug (MIC less than 4 micrograms/ml). Escherichia coli, Klebsiella pneumoniae and Salmonella sp. showed intermediate susceptibility (MIC 4-16 micrograms/ml), while all isolates of Corynebacterium (Rhodococcus) equi, Enterobacter cloacae and Pseudomonas aeruginosa proved to be highly resistant to cefadroxil (MIC greater than 128 micrograms/ml).


Challenges and opportunities with glycogen synthase kinase-3 inhibitors for insulin resistance and Type 2 diabetes treatment.
The role of the serine/threonine protein kinase, glycogen synthase kinase-3 (GSK-3), in attenuating the insulin signalling pathway has led to the concept that inhibition of GSK-3 may have therapeutic benefits in the treatment of insulin resistance and Type 2 diabetes. Indeed, various selective GSK-3 inhibitors have been developed recently and have proven to promote insulin-like effects and to act as insulin sensitisers in both in vitro and in vivo systems. GSK-3 inhibition may thus present a new, effective approach for the treatment of insulin resistance and Type 2 diabetes. This review describes the qualifications of GSK-3 as a novel drug-discovery target for Type 2 diabetes and discusses the strategies and challenges in developing small-molecule inhibitors for this important protein kinase.


Metastatic adenocarcinoma of mandible: in search of the primary.
We present here a case of a 64-year-old female patient who reported with a complaint of pain in the chin region of 3-month duration. The radiographs revealed an irregular radiolucent lesion in the anterior part of the mandible. An incisional biopsy was carried out and the microscopy showed features of adenocarcinoma. Suspecting a metastatic lesion, the patient was then sent for whole body examination which showed a mass in the lungs. A whole body scan also showed metastasis to other organs such as kidney, gallbladder and vertebrae. This case is unusual in that the patient complained only of a vague jaw pain with no other systemic symptoms even in the presence of widespread disease.


Comparison of 80 × 112-Gb/s PDM-QPSK system performance over large effective area fiber and standard SMF with Raman amplification.
We experimentally investigate the transmission performance of 80 × 112-Gb/s polarization-division-multiplexed quadrature phase shift keying (PDM-QPSK) signals over large effective area fiber and standard single mode fiber (SSMF) links with Raman amplification. The large effective area fiber offers higher optimum launch power and longer reach than SSMF. The maximum reach of 5200-km is obtained using large effective area fiber. The Gaussian noise (GN) model is explored to fit with experimental data for optimum power.


The scale of repeat prescribing.
Repeat prescribing has long been seen as a potential cause of poor clinical care, despite its obvious advantage to both doctors and patients. Previous studies have had no common definition of the term, and have been small in scale, but it is clear that repeat prescribing has increased over the past 25 years with a recent acceleration due to computerization. Managing the process has become more important as the scale has increased. A computer-related standard definition would provide linkage with other information held on the practice computer about the recipients. Using aggregated practice data the current national picture could be ascertained for comparison with that of individual practices. At practice level it will be less important simply to know the scale of repeat prescribing than to make analyses of repeat prescribing of particular drug groups, and of the age and sex groups of the recipients. This could provide a valuable basis for improving clinical care. To estimate the present scale of repeat prescribing-overall, for specific age-sex groups, and for some specific drug groups; to provide a much needed standard definition of repeat prescribing, now inevitably related to computer procedures; and to show how clinically valuable audits might be simply generated as reports by a practice computer. Repeat prescriptions were defined as those printed by a practice computer from its repeat prescribing program over a period of one year. Prescribing data for a year, with demographic details of the patients involved, were obtained for 115 practices from the IMS MediPlus database. These practices had 750390 patients and issued 5.82 million prescriptions during the year. Analyses were made of the overall percentages of items and costs due to repeats; the percentage of patients receiving repeats, by age and sex; the percentage receiving repeats, by age and sex, in areas of particular concern; and percentage repeat prescribing in 46 drug groups. No differences were found between fundholding and non-fundholding practices, or between dispensing and non-dispensing practices. The ratio of acute to repeat prescriptions in the practices was stable over four years. Repeats accounted for 75% of all items and 81% of prescribing costs; 48.4% of all patients were receiving a repeat prescription. Many drugs, including hypnotics, were given almost entirely as repeats. The percentage of repeats increased with patients' age, from 36% in the 0-4 year age group to more than 90% for patients aged 85 and over. It was higher overall for males than for females, though this relationship did not hold for older patients. This study gives the best available national picture of the use of repeat prescribing. The definition employed does not allow any direct conclusions to be drawn about whether the patients involved were being given adequate clinical care, but it permits analyses at practice level that can indicate where special attention may be required. It could usefully be adopted as the much-needed standard definition.


Auditory cueing strategy for stride length and cadence modification: a feasibility study with healthy adults.
People with Parkinson's Disease experience gait impairments that significantly impact their quality of life. Visual, auditory, and tactile cues can alleviate gait impairments, but they can become less effective due to the progressive nature of the disease and changes in people's motor capability. In this study, we develop a human-in-the-loop (HIL) framework that monitors two key gait parameters, stride length and cadence, and continuously learns a person-specific model of how the parameters change in response to the feedback. The model is then used in an optimization algorithm to improve the gait parameters. This feasibility study examines whether auditory cues can be used to influence stride length in people without gait impairments. The results demonstrate the benefits of the HIL framework in maintaining people's stride length in the presence of a secondary task.Clinical relevance- This paper proposes a gait rehabilitation framework that provides a personalized cueing strategy based on the person's real-time response to cues. The proposed approach has potential application to people with Parkinson's Disease.


Comparison of Nanofiltration with Reverse Osmosis in Reclaiming Tertiary Treated Municipal Wastewater for Irrigation Purposes.
This study compares the performance of nanofiltration (NF) and reverse osmosis (RO) for the reclamation of ultrafiltered municipal wastewater for irrigation of food crops. RO and NF technologies were evaluated at different applied pressures; the performance of each technology was evaluated in terms of water flux, recovery rate, specific energy consumption and quality of permeate. It was found that the permeate from the reverse osmosis (RO) process complied with Food and Agriculture Organization (FAO) standards at pressures applied between 10 and 18 bar. At an applied pressure of 20 bar, the permeate quality did not comply with irrigation water standards in terms of chloride, sodium and calcium concentration. It was found that nanofiltration process was not suitable for the reclamation of wastewater as the concentration of chloride, sodium and calcium exceeded the allowable limits at all applied pressures. In the reverse osmosis process, the highest recovery rate was 36%, which was achieved at a pressure of 16 bar. The specific energy consumption at this applied pressure was 0.56 kWh/m3. The lowest specific energy of 0.46 kWh/m3 was achieved at an applied pressure of 12 bar with a water recovery rate of 32.7%.


Molecular epidemiology of respiratory syncytial virus in Kilifi district, Kenya.
Respiratory syncytial virus (RSV) causes significant burden of disease during infancy and childhood. This study examined the genetic relatedness of RSV positive samples from child inpatients and outpatients and a birth cohort from a rural coastal district of Kenya and also the distribution of strains between these three groups. Clinical samples were collected over a 4-year period in Kilifi District, Kenya from community and hospital surveillance. Three hundred ninety seven of 1,044 nasal specimens from children (under 5 years old) attending Kilifi District Hospital, and from community-monitored infants, were positive for RSV by multiplex RT-PCR. Of these, 376 samples were analysed further by restriction fragment length polymorphisms (RFLP) of the nucleocapsid (N) and attachment (G) protein genes. The G gene was sequenced for 109 samples and phylogenetic analysis carried out. The group A samples from Kilifi fell into two clusters based on G gene sequences, while only one group B cluster was observed. One RSV-B sample from 2003 demonstrated the presence of a 60-nucleotide duplication within the G gene, clustering with similar isolates from Buenos Aries from 1999. All had similar sequences to isolates from the UK, USA, Spain, or Uruguay. The Kilifi District samples showed greater than 97% homology to isolates from South Africa and Mozambique and 91-94% homology to isolates from The Gambia. Samples from different sources, clearly differing in disease severity, did not differ in genotype characteristics, suggesting that disease causing variants are a general reflection of infections within this community.


Identification of quantitative trait loci affecting cattle temperament.
In addition to its potential contribution to improving animal welfare, the study of the genetics of cattle behavior may provide more general insights into the genetic control of such complex traits. We carried out a genome scan in a Holstein x Charolais cross cattle population to identify quantitative trait loci (QTL) influencing temperament-related traits. Individuals belonging to the second-generation of this population (F(2) and backcross individuals) were subjected to 2 behavioral tests. The flight from feeder (FF) test measured the distance at which the animal moved away from an approaching human observer, whereas the social separation (SS) test categorized different activities which the animal engaged in when removed from its penmates. The entire population was genotyped with 165 microsatellite markers. A regression interval mapping analysis identified 29 regions exceeding the 5% chromosome-wide significance level, which individually explained a relatively small fraction of the phenotypic variance of the traits (from 3.8% to 8.4%). One of the significant associations influencing an FF test trait on chromosome 29 reached the 5% genome-wide significance level. Eight other QTL, all associated with an SS test trait, reached the 1% chromosome-wide significance level. The location of some QTL coincided with other previously reported temperament QTL in cattle, whereas those that are reported for the first time here may represent general loci controlling temperament differences between cattle breeds. No overlapping QTL were identified for the traits measured by the 2 different tests, supporting the hypothesis that different genetic factors influence behavioral responses to different situations.


Down-regulation of microRNA-23b aggravates LPS-induced inflammatory injury in chondrogenic ATDC5 cells by targeting PDCD4.
Osteoarthritis (OA), characterized by degradation of articular cartilage, is a leading cause of disability. As the only cell type present in cartilage, chondrocytes play curial roles in the progression of OA. In our study, we aimed to explore the roles of miR-23b in the lipopolysaccharide (LPS)-induced inflammatory injury. LPS-induced cell injury of ATDC5 cells was evaluated by the loss of cell viability, enhancement of cell apoptosis, alteration of apoptosis-associated proteins, and release of inflammatory cytokines. Then, miR-23b level after LPS treatment was assessed by qRT-PCR. Next, the effects of aberrantly expressed miR-23b on the LPS-induced inflammatory injury were explored. The possible target genes of miR-23b were virtually screened by informatics and verified by luciferase assay. Subsequently, whether miR-23b functioned through regulating the target gene was validated. The involved signaling pathways were investigated finally. Cell viability was decreased but cell apoptosis, as well as release of inflammatory cytokines, was enhanced by LPS treatment. MiR-23b was down-regulated by LPS and its overexpression alleviated LPS-induced inflammatory injury. PDCD4, negatively regulated by miR-23b expression, was verified as a target gene of miR-23b. Following experiments showed miR-23b alleviated LPS-induced cell injury through down-regulating PDCD4 expression. Phosphorylated levels of key kinases in the NF-κB pathway, as well as expressions of key kinases in the Notch pathways, were increased by PDCD4 overexpression. MiR-23b was down-regulated after LPS treatment, and its overexpression ameliorated LPS-induced inflammatory injury in ATDC5 cells by targeting PDCD4, which could activate the NF-κB/Notch pathways.


Life history of the predatory mites Neoseiulus paspalivorus and Proctolaelaps bickleyi, candidates for biological control of Aceria guerreronis.
The eriophyoid mite Aceria guerreronis Keifer (Eriophyidae), commonly called the coconut mite, is a key pest of coconut fruits. Surveys conducted on coconut palms in Brazil revealed the predatory mites Neoseiulus paspalivorus DeLeon (Phytoseiidae) and Proctolaelaps bickleyi Bram (Ascidae) as the most commonly associated natural enemies of A. guerreronis on coconut fruits. However, virtually nothing is known about the life history of these two predators. We conducted laboratory experiments at 25 +/- 0.1 degrees C, 70-90% RH and 12:12 h L:D photoperiod to determine the life history characteristics of the two predatory mites when feeding on A. guerreronis and other potential food sources present on coconut fruits such as Steneotarsonemus furcatus DeLeon (Tarsonemidae), coconut pollen and the fungus Rhizopus cf. stolonifer Lind (Mucoraceae). In addition, the two-spotted spider mite Tetranychus urticae Koch (Tetranychidae) was tested for its suitability as prey. Both predators, N. paspalivorus and P. bickleyi, thrived on A. guerreronis as primary food source resulting in shorter developmental time (5.6 and 4.4 days, respectively), higher oviposition rate (1.7 and 7.0 eggs/female/day, respectively) and higher intrinsic rate of increase (0.232 and 0.489 per female/day, respectively) than on any other diet but were unable to develop or lay eggs when fed T. urticae. Coconut pollen and S. furcatus were adequate alternative food sources for N. paspalivorus and Rhizopus for P. bickleyi. We discuss the relevance of our findings for natural and biological control of the coconut mite A. guerreronis.


Differential synapse formation and neurite outgrowth at two branches of the metacerebral cell of Aplysia in dissociated cell culture.
The metacerebral cell (MCC) of Aplysia californica was isolated with its bifurcate axon from the cerebral ganglion and maintained in vitro under three conditions: (a) with no targets, (b) with identified buccal ganglion neurons B1 or B2 placed near the stump of the large diameter cerebral-buccal connective (CBC) branch, and (c) with B1 or B2 placed near the stump of the small diameter posterior lip nerve (PLN) branch. After 5 days in culture, the two branches differed significantly in the formation of chemical connections and in the extent of neurite outgrowth. Chemical connections characteristic of MCC-B1(B2) connections in vivo were observed in more than 90% of the cultures in which the buccal neuron was contacted by neurites emerging from the CBC branch, but in only 20% of the cultures in which the buccal neuron was contacted by neurites extending from the PLN branch. Neurite outgrowth from the CBC stump was always greater than growth from the PLN and was not affected significantly by the presence of a buccal neuron target at either branch. In contrast, neurite outgrowth from the PLN decreased significantly when the target was contacted by neurites from the CBC branch. These results suggest that two branches of a single neuron can differ in their capacities to form chemical connections. In addition, the two branches show differential growth as a result of target interaction at one of the branches. This simple in vitro system may therefore be useful in exploring the ways in which individual neurons control neurite extension from different branches as they seek to form chemical connections with their targets.


Absence of host-site influence on angiogenesis, blood flow, and permeability in transplanted RG-2 gliomas.
The host site is believed to regulate tumor angiogenesis, which could result in site-dependent drug delivery parameters, greatly affecting experimental tumor research. In RG-2 rat gliomas we measured cellular proliferation; cell cycle time was the same for RG-2 cells in brain and s.c. tumors (25 h), and was the same for endothelial cells in these tumors (46 h). We measured the transcapillary transfer constant (K) of alpha-aminoisobutyric acid and blood flow (F) with iodoantipyrine in RG-2 gliomas transplanted into brain, liver, kidney, muscle, s.c. tissue, and into the abdominal cavity. Data was evaluated by quantitative autoradiography and direct tissue sampling. The variation of F (12.6-84.0 ml/g/min) and K (26.1-49.2 microl/g/min) in RG-2 tumors in the different host sites was less than in surrounding tumor-free tissue (F = 20-1500 ml/g/min and K = 1.6-700 microl/g/min). In contrast to other models, RG-2 does not result in tumors with host site-dependent behavior. The RG-2 tumor cells appear to participate in, if not dominate, the angiogenesis process regardless of the host site. Values of F and K were more dependent on tumor topography (center versus periphery) and local histological features (necrosis versus viable tumor) than host site. We believe that the methods used for data acquisition may introduce as much variability in Results as the tumors themselves and that to better understand how tumor angiogenesis affects the vascular phenotype, comparative studies are needed to validate the results obtained with newer methodologies.


The challenge of solvent substitution in coatings.
A brief outline of the role of solvents in surface coatings is given, to provide the background to the problems encountered in solvent substitution. The direct substitution of one solvent by another is relatively straightforward. Computer programs greatly simplify the process. However, the real challenge lies with the reduction in solvent use, through the development of low solvent coatings. While this is a process that has been going on for many years, there is no doubt that recent regulatory pressures regarding both occupational health and environmental standards have made it the single most important challenge for the industry. Examples are discussed of the development of high solids, low solvent industrial coatings as well as water borne industrial coatings. Although there has been rapid progress in recent years, there is no prospect in sight for the complete elimination of solvents. Good workplace hygiene must therefore remain a key element in the safe use of coatings.


Prematurity and school readiness in a nationally representative sample of Australian children: does typically occurring preschool moderate the relationship?
This study aims to examine the relationship between indicators of prematurity and children's cognitive and behavioral school readiness in a nationally representative sample and to investigate whether typically occurring preschool enrollment moderates this relationship, particularly for children from disadvantaged families in Australia. The Longitudinal Study of Australian Children is a nationally representative prospective sample of two cohorts of children with sequentially obtained indicators of child health and developmental outcomes. We analyzed information on 8060 children aged 4-5 years who had complete data on birth weight, gestational age, prenatal risks, social factors, and cognitive and behavioral outcomes of school readiness. Multivariate regressions were used to relate three indicators of prematurity (low birth weight, preterm birth, and small for gestational age) to cognitive and behavioral school readiness. Children born preterm, small for gestational age, or with low birth weight have significantly lower cognitive school readiness after controlling for social factors and prenatal risks. None of the premature indicators were associated with behavioral school readiness. All children benefited from attending preschool. Yet, preschool enrollment did not moderate the relationship between prematurity and school readiness. The only exception is for small for gestational age survivors with low educated mothers. Preschool enrollment was associated with an increase in cognitive school readiness skills. Prematurity was associated with lower cognitive school readiness skills. Typical occurring preschool did not eliminate this association. Findings suggest that simply expanding the preschool enrollment is inadequate to address the developmental needs of premature children from disadvantaged backgrounds.


Predicting reaction performance in C-N cross-coupling using machine learning.
Machine learning methods are becoming integral to scientific inquiry in numerous disciplines. We demonstrated that machine learning can be used to predict the performance of a synthetic reaction in multidimensional chemical space using data obtained via high-throughput experimentation. We created scripts to compute and extract atomic, molecular, and vibrational descriptors for the components of a palladium-catalyzed Buchwald-Hartwig cross-coupling of aryl halides with 4-methylaniline in the presence of various potentially inhibitory additives. Using these descriptors as inputs and reaction yield as output, we showed that a random forest algorithm provides significantly improved predictive performance over linear regression analysis. The random forest model was also successfully applied to sparse training sets and out-of-sample prediction, suggesting its value in facilitating adoption of synthetic methodology.


The relationship of asprosin with β-hydroxybutyric acid and postpartum disorders in cows.
The aim of the present study was to determine asprosin levels in cows, the relationship of this hormone with postpartum disorders and β-hydroxybutyric acid, and also the potential of asprosin to be a marker for postpartum diseases. The study was designed as a two-stage trial. In the first stage, blood asprosin and β-hydroxybutyric acid levels of 20 healthy Simmental cows aged 3-4 years were measured at the time of calving, and on days 3, 6, 9, 12 and 15 postpartum. In the second stage, 200 cows were divided into two groups: (1) healthy (n = 100) and (2) diseased (placental retention, hypocalcaemia, metritis, lameness, abomasal displacement, mastitis; n = 100); asprosin and β-hydroxybutyric acid concentrations of the blood were assessed on day 15 postpartum. In conclusion, the asprosin level was found to be at measurable levels in cows, and a negative correlation with β-hydroxybutyric acid was found. According to these findings, the data obtained from this study could be used for the prevention, control and treatment of some postpartum disorders associated with ketosis and for developing novel hypotheses concerning the actions of this hormone. It was concluded that further studies are required to reveal the associations between asprosin and postpartum disorders.


The ionization energy of the vinyl radical: a Mexican standoff with a happy ending.
The Active Thermochemical Tables (ATcT) yield 8.477 ± 0.007 eV as the ionization energy of the vinyl radical, which agrees with wave function theory results. The photoionization spectrum yielded 8.59 ± 0.03 eV, while the photoelectron spectrum yielded 8.25 ± 0.05 eV for the adiabatic ionization energy. In order to reconcile these contradictory measurements, we produced the vinyl radical by flash pyrolysis of divinyl sulfone, and recorded its photoionization (PI) and threshold photoelectron (TPE) spectrum in the 8.10-10.45 eV photon energy range at the Swiss Light Source for the first time using double imaging photoelectron photoion coincidence (i2PEPICO) spectroscopy. In contrast to the photoelectron spectrum, the ground state TPES band exhibits a weak vibrational structure that correlates with strong peaks in the photoionization spectrum and is assigned to transitions to autoionizing Rydberg states converging to the 3A'' cation state. While, at 8.462 eV, our calculations of the acetylene proton affinity confirm the vinyl adiabatic ionization energy from the ATcT, our experimental data agree with the literature photoelectron and photoionization spectra. The discrepancy is explained by negligible Franck-Condon factors for the origin transition from the bent Y-shaped neutral minimum to the bridge-shaped ionic minimum. We propose that the photoionization signal can instead be assigned in its entirety to three transitions: (i) straight Y-shaped C2H3‡→ bridged C2H3+, (ii) straight Y-shaped C2H3‡→ straight Y-shaped C2H3+ and (iii) bent Y-shaped C2H3→ straight Y-shaped C2H3+, tentatively assigned to the observed photoionization onsets at 8.25, 8.43, and 8.60 eV, respectively.


Feasibility, Acceptability, and Usability of Physiology and Emotion Monitoring in Adults and Children Using the Novel Time2Feel Smartphone Application.
The present study tests the feasibility, acceptability, and utility of the novel smartphone application-Time2Feel-to monitor family members' emotional experiences, at the experiential and physiological level, and their context. To our knowledge, Time2Feel is the first of its kind, having the capability to monitor multiple members' emotional experiences simultaneously and survey users' emotional experiences when experiencing an increase in physiological arousal. In this study, a total of 44 parents and children used Time2Feel along with the Empatica E4 wrist-wearable device for 10 days. Engagement rates were within the acceptable range and consistent with previous work using experience sampling methods. Perceived ease of use and satisfaction fell mostly in the moderate range, with users reporting challenges with connectivity. We further discuss how addressing connectivity would increase acceptability. Finally, Time2Feel was successful at identifying physiological deviations in electrodermal activity for parents and children alike, and even though responses to those deviation-generated surveys were largely consistent with random survey responses, some differences were noted for mothers and fathers. We discuss the implications of using Time2Feel for understanding families' emotional and stressful experiences day-to-day.


A systematic exploration of the influence of the protein stability on amyloid fibril formation in vitro.
There are a number of diseases in which normally soluble proteins associate into regular, insoluble amyloid fibrils. The development of in vitro model systems in which detailed structural, kinetic, and thermodynamic characterization are feasible is of critical importance to our understanding of the amyloid fibril phenomenon. The formation of amyloid fibrils by proteins that are not associated with disease has been recently described, suggesting that this may be a common property of many proteins and not only of the few proteins associated with amyloidoses. The B1 Ig-binding domain of protein G (beta1) is an extremely well-characterized model system. We have found that under certain experimental conditions, some variants of beta1 form fibrils with high reproducibility. By controlling the stability of the protein-either by mutations or by changing experimental conditions-we are able to modulate the ability of the protein to form fibrils. For all of the variants, we find that the key requirement for fibril formation is to choose conditions in which the population of intermediate conformations present during the unfolding transition is maximized.


Infectious gastroenteritis in Norfolk Island and recovery of viruses from drinking water.
A high incidence of gastroenteritis in both islanders and tourists has been recorded in recent years on Norfolk Island--a popular tourist resort for Australians and New Zealanders. No bacterial cause has been found. However, electron microscopic examination of 28 faecal specimens revealed viruses associated with gastroenteritis in 21 (75%). No viruses were isolated in cell cultures. Bore water is used for drinking purposes on the island and 32 samples from 15 bores were examined for viruses by electron microscopy and culture as well as for bacterial contamination. Seven polioviruses (all type 1 vaccine strain) and adenoviruses 1 and 5 were isolated in cell cultures. In addition one rotavirus, one adenovirus and two small round viruses were detected by electron microscopy. Six of 21 samples tested showed unacceptably high levels of bacteria for drinking water. The deep ground water has apparently become contaminated with sewage effluent and is almost certainly the main cause of the high level of gastroenteritis on the island.


Effects of a monoclonal antibody raised against nerve growth factor on skeletal pain and bone healing after fracture of the C57BL/6J mouse femur.
A closed femur fracture pain model was developed in the C57BL/6J mouse. One day after fracture, a monoclonal antibody raised against nerve growth factor (anti-NGF) was delivered intraperitoneally and resulted in a reduction in fracture pain-related behaviors of approximately 50%. Anti-NGF therapy did not interfere with bone healing as assessed by mechanical testing and histomorphometric analysis. Current therapies to treat skeletal fracture pain are limited. This is because of the side effect profile of available analgesics and the scarcity of animal models that can be used to understand the mechanisms that drive this pain. Whereas previous studies have shown that mineralized bone, marrow, and periosteum are innervated by sensory and sympathetic fibers, it is not understood how skeletal pain is generated and maintained even in common conditions such as osteoarthritis, low back pain, or fracture. In this study, we characterized the pain-related behaviors after a closed femur fracture in the C57BL/6J mouse. Additionally, we assessed the effect of a monoclonal antibody that binds to and sequesters nerve growth factor (anti-NGF) on pain-related behaviors and bone healing (mechanical properties and histomorphometric analysis) after fracture. Administration of anti-NGF therapy (10 mg/kg, days 1, 6, and 11 after fracture) resulted in a reduction of fracture pain-related behaviors of approximately 50%. Attenuation of fracture pain was evident as early as 24 h after the initial dosing and remained efficacious throughout the course of fracture pain. Anti-NGF therapy did not modify biomechanical properties of the femur or histomorphometric indices of bone healing. These findings suggest that therapies that target NGF or its cognate receptor(s) may be effective in attenuating nonmalignant fracture pain without interfering with bone healing.


Effect of exercise on serum estrogens in postmenopausal women: a 12-month randomized clinical trial.
Elevated circulating estrogens and a sedentary lifestyle increase risk for breast cancer. The effect of exercise on circulating estrogens in sedentary postmenopausal women is unknown. The objective of this study was to examine the effects of a 12-month moderate-intensity exercise intervention on serum estrogens. We randomly assigned 173 sedentary, overweight (body mass index > 24.0 kg/m(2), body fat > 33%), postmenopausal women, ages 50-75 years, not using hormone therapy, living in the Seattle, Washington, area for the next year, and willing to be randomly assigned to an exercise intervention or stretching control group. The exercise intervention included facility and home-based exercise (45 min, 5 days/week moderate intensity sports/recreational exercise). A total of 170 (98.3%) women completed the study with exercisers averaging 171 min/week of exercise. After 3 months, exercisers experienced declines in estrone, estradiol, and free estradiol of 3.8, 7.7, and 8.2%, respectively, versus no change or increased concentrations in controls (P = 0.03, 0.07, and 0.02, respectively). At 12 months, the direction of effect remained the same, although the differences were no longer statistically significant. The effect was limited to women who lost body fat: women whose percentage of body fat [by dual energy x-ray absortiometry (DEXA)] decreased by >/==" BORDER="0">2% had statistically significant (comparing exercisers versus controls) decreases at 12 months of 11.9, 13.7, and 16.7% for serum estrone, estradiol, and free estradiol, respectively. We concluded that a 12-month moderate-intensity exercise intervention in postmenopausal women resulted in significant decreases in serum estrogens. The association between increased physical activity and reduced risk for postmenopausal breast cancer may be partly explained by effects on serum estrogens.


Adsorption of organic matter on titanium surfaces with nano- and micro-scale roughness studied with the electrochemical quartz crystal microbalance dissipation technique.
Adsorption of calf serum organic matter from a phosphate-buffered solution was studied using the electrochemical quartz crystal microbalance with additional dissipation measurements. Two types of crystal surfaces were used: one rough with micrometer-range surface features and one with roughness in the low nanometer range. The results showed that the adsorption of the organic material was about 1.5 orders of magnitude larger on the rough surface and almost independent of serum concentration in the electrolyte. The adsorption rates were found to increase with increasing serum concentration. For rough crystals, the adsorption kinetics were interpreted with the Johnson-Mehl-Avrami-Kolmogorov model, indicating an initial growth phase according to the tn-law, followed by a slower growth as the nucleation sites fill up. This study suggests that specific surface sites are critical to promote adsorption of proteins on a titanium surface.


Identifying genetic changes associated with tumor progression in squamous cell carcinoma.
Tumor behavior is the result of specific genetic changes that alter gene expression. From our cytogenetic studies chromosome 18 loss emerged as a common genetic change in squamous carcinoma cell lines. In this report we summarize data that link loss of 18 to tumor progression and reduced survival, indicating that one or more tumor suppressor gene(s) are located on this chromosome. Tumors grown in vitro were karyotyped either as short-term or permanent cultures. Loss of chromosome 18 was measured by karyotype, decreased frequency of heterozygosity at the DCC locus, and loss of heterozygosity (LOH) for microsatellite repeat polymorphisms (MSRP). Loss of any part of chromosome 18 was observed in approximately 63% of cultured tumors. Primary and secondary tumors from the same individuals sometimes differed in loss of 18 indicating that this genetic change is associated with tumor progression. Heterozygosity for DCC was present in only 3/19 cultured SCC (16%), compared with 68% (11/16) of blood samples from unrelated donors, which is consistent with LOH in roughly one half of the cases. Of 4 informative cases with normal and tumor tissue, LOH was observed in 2. Microsatellite analysis also shows loss of 18q in 55% of fresh tumors. Analysis of tumor tissue and cell lines from the same patient gave identical results. There was an excess of deaths from cancer in the group with 18 loss (20/25) when compared with the group without (5/15). Loss of chromosome 18 appears to be a marker of tumor progression in SCC. It is likely that mutation affecting DCC or another gene on 18 affects tumor growth or spread, leading to more rapid progression and reduced survival.


Intravaginal gel for staging of female pelvic cancers--preliminary report of safety, distention, and gel-mucosal contrast during magnetic resonance examination.
To more fully outline cervical and vaginal contours and distend the vagina, we have filled the vagina with sterile water-based gel before the magnetic resonance (MR) examination. The technique is similar to that used for defecating MR proctography and other MR examinations, but has not been well described for MR imaging of female pelvic cancer. We present our preliminary clinical experience, including a review of safety imaging characteristics and maintenance of the distention during the examination.


De novo lumen formation and elongation in the developing nephron: a central role for afadin in apical polarity.
A fundamental process in biology is the de novo formation and morphogenesis of polarized tubules. Although these processes are essential for the formation of multiple metazoan organ systems, little is known about the molecular mechanisms that regulate them. In this study, we have characterized several steps in tubule formation and morphogenesis using the mouse kidney as a model system. We report that kidney mesenchymal cells contain discrete Par3-expressing membrane microdomains that become restricted to an apical domain, coinciding with lumen formation. Once lumen formation has been initiated, elongation occurs by simultaneous extension and additional de novo lumen generation. We demonstrate that lumen formation and elongation require afadin, a nectin adaptor protein implicated in adherens junction formation. Mice that lack afadin in nephron precursors show evidence of Par3-expressing membrane microdomains, but fail to develop normal apical-basal polarity and generate a continuous lumen. Absence of afadin led to delayed and diminished integration of nectin complexes and failure to recruit R-cadherin. Furthermore, we demonstrate that afadin is required for Par complex formation. Together, these results suggest that afadin acts upstream of the Par complex to regulate the integration and/or coalescence of membrane microdomains, thereby establishing apical-basal polarity and lumen formation/elongation during kidney tubulogenesis.


The topography of fibrous scaffolds modulates the paracrine function of Ad-MSCs in the regeneration of skin tissues.
Injuries to the skin are common in daily life, and a certain type or size of defect is not easily restored using conventional dressings or naturally. The repair of these defects requires restoration of function in regenerated tissues. In this study, a tissue engineered skin was designed and fabricated using a bio-3D printing system. Polycaprolactone and bacterial cellulose comprised the scaffold, due to their excellent biocompatibility and multifunctionality. Adipose-derived mesenchymal stem cells (Ad-MSCs) were seeded onto the scaffold to functionalize it as an artificial skin. The finished artificial skin had mechanical properties similar to that of natural skin, and its fibrous structure providing a unique micro-environment that could regulate the paracrine function of the Ad-MSCs. This effect could be greatly increased by changes in the characteristics of the biomaterials. The artificial skin exhibited high biological activity, strong induction of cell recruitment, migration, growth and up-regulation of gene expression of relevant factors, resulting in excellent wound healing characteristics. This study clarified novel design aspects of cell-material interactions in which the topographical characteristics of materials can be further developed to establish cell signaling or communication networks that take advantage of the paracrine actions of Ad-MSCs to promote specific tissue regeneration or repair characteristics.


Functional Recovery After Postpartum Psychosis: A Prospective Longitudinal Study.
Postpartum psychosis is an acute and severe mood disorder. Although the prognosis is generally good, postpartum psychosis is a highly stressful life-event presumed to have a major impact on functioning and well-being beyond the acute stage of the illness. We studied functional recovery, including psychosocial functioning and the presence of psychological distress, in patients with a recent diagnosis of postpartum psychosis. Seventy-eight patients with postpartum psychosis consecutively admitted for inpatient hospitalization between 2005 and 2011 were assessed 9 months postpartum. Included were patients with DSM-IV-TR diagnoses of psychotic disorder not otherwise specified, brief psychotic disorder, or mood disorder with psychotic features, each requiring the additional specifier "with postpartum onset." Functioning was assessed in 4 domains by the Longitudinal Interval Follow-up Evaluation-Range of Impaired Functioning Tool (LIFE-RIFT). Symptomatology was measured by the Brief Symptom Inventory and compared to a matched population-based cohort. Nine months postpartum, 74% (58/78) of women with postpartum psychosis reported good functioning on the domains of work, interpersonal relations, recreation, and global satisfaction. Moreover, 88% (69/78) of patients with postpartum psychosis had resumed their premorbid employment and household responsibilities. Compared to the general population, patients with postpartum psychosis reported a higher burden of depression and anxiety (effect sizes r ≤ 0.14). Patients who had a relapse episode (18%) experienced considerable functional impairments across several domains. Nine months postpartum, the majority of patients with postpartum psychosis reported good functional recovery. Our relatively improved functional outcomes compared to nonpostpartum onset could be attributed to the postpartum onset and/or more favorable risk factor profile.


Adenylate kinase of Escherichia coli: evidence for a functional interaction in phospholipid synthesis.
Previous genetic and biochemical experiments have suggested that the adenylate kinase of Escherichia coli may be directly involved in phospholipid synthesis through formation of a complex with sn-glycerol-3-phosphate acyltransferase, the membrane-bound enzyme that catalyzes the first step in phospholipid synthesis. In this paper we report direct experiments to test this hypothesis. A mutation within the adenylate kinase structural gene is described that results in a temperature-sensitive phospholipid synthesis (assayed in vivo) and a temperature-sensitive acyltransferase. The adenylate kinase activity of this strain is only minimally altered either in vitro or [as assayed by adenosine 5'-triphosphate (ATP) levels] in vivo. This result demonstrates that the inhibition of phospholipid synthesis is not the result of reduced ATP levels. We report the purification of E. coli adenylate kinase to homogeneity; and find that the addition of homogeneous wild-type adenylate kinase to membranes containing a mutationally altered temperature-sensitive acyltransferase results in thermal stabilization of the acyltransferase activity. Ovalbumin has no such protective effect. Purified E. coli inner membranes contain several proteins that are precipitated by addition of anti adenylate kinase antibody to detergent-solubilized membranes.


Optical magnetic field mapping using a subwavelength aperture.
Local distribution of the optical magnetic field is a critical parameter in developing materials with artificially engineered optical properties. Optical magnetic field characterization in nano-scale remains a challenge, because of the weak matter-optical magnetic field interactions. Here, we demonstrate an experimental visualization of the optical magnetic field profiles by raster scanning circular apertures in metal film or in a conical probe. Optical magnetic fields of surface plasmon polaritons and radially polarized beam are visualized by measuring the transmission through metallic apertures, in excellent agreements with theoretical predictions. Our results show that Bethe-Bouwkamp aperture can be used in visualizing optical magnetic field profiles.


The characterization of micro-organisms by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
A variety of gram-positive and gram-negative intact bacterial cells have been analysed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and shown to provide fingerprint mass spectra with discrete peaks being observed over the mass range from 3 to 40 kDa. The spectra show both more peaks and peaks at a higher mass/charge ratio than have hitherto been reported for these micro-organisms and would appear to provide a profile of cellular proteinaceous material. The spectra are shown to be reproducible over variable time periods of up to three months and factors affecting reproducibility are discussed. The procedure, which requires minimal sample preparation, yields results in 30-40 minutes and allows visual identification of species- and strain-specific biomarkers for the characterization of the organisms. The importance of accurately defining sample preparation methodologies is central to the ability of the technique to generate reliable and reproducible data.


[Computer tomography in the diagnosis of retroperitoneal nonorganic tumors].
The results of a follow-up of 128 patients with retroperitoneal nonorganic tumors showed that among diagnostic methods computerized tomography (CT) proved to be a valuable method in the recognition of nonorganic tumors of the retroperitoneal space permitting one to detect neoplasms in 92.6%. In spite of the fact that the main bulk of retroperitoneal nonorganic tumors had various images on computerized tomograms this method provided an opportunity to judge of the nature of nonorganic neoplasms on the basis of correlation of their structural elements, the prevalence of this or that tissue and the expression of disintegration processes. Diagnostic errors in CT of the patients with retroperitoneal nonorganic tumors were associated with objective difficulties determined by the peculiarities of interrelationships of a growing tumor with the adjacent organs.


Dietary supplements for dysmenorrhoea.
Dysmenorrhoea refers to painful menstrual cramps and is a common gynaecological complaint. Conventional treatments include non-steroidal anti-inflammatory drugs (NSAIDs) and oral contraceptive pills (OCPs), which both reduce myometrial activity (contractions of the uterus). A suggested alternative approach is dietary supplements. We used the term 'dietary supplement' to include herbs or other botanical, vitamins, minerals, enzymes, and amino acids. We excluded traditional Chinese medicines. To determine the efficacy and safety of dietary supplements for treating dysmenorrhoea. We searched sources including the Cochrane Gynaecology and Fertility Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, AMED, PsycINFO (all from inception to 23 March 2015), trial registries, and the reference lists of relevant articles. We included randomised controlled trials (RCTs) of dietary supplements for moderate or severe primary or secondary dysmenorrhoea. We excluded studies of women with an intrauterine device. Eligible comparators were other dietary supplements, placebo, no treatment, or conventional analgesia. Two review authors independently performed study selection, performed data extraction and assessed the risk of bias in the included trials. The primary outcomes were pain intensity and adverse effects. We used a fixed-effect model to calculate odds ratios (ORs) for dichotomous data, and mean differences (MDs) or standardised mean differences (SMDs) for continuous data, with 95% confidence intervals (CIs). We presented data that were unsuitable for analysis either descriptively or in additional tables. We assessed the quality of the evidence using Grading of Recommendations Assessment, Development and Evaluation (GRADE) methods. We included 27 RCTs (3101 women). Most included studies were conducted amongst cohorts of students with primary dysmenorrhoea in their late teens or early twenties. Twenty-two studies were conducted in Iran and the rest were performed in other middle-income countries. Only one study addressed secondary dysmenorrhoea. Interventions included 12 different herbal medicines (German chamomile (Matricaria chamomilla, M recutita, Chamomilla recutita), cinnamon (Cinnamomum zeylanicum, C. verum), Damask rose (Rosa damascena), dill (Anethum graveolens), fennel (Foeniculum vulgare), fenugreek (Trigonella foenum-graecum), ginger (Zingiber officinale), guava (Psidium guajava), rhubarb (Rheum emodi), uzara (Xysmalobium undulatum), valerian (Valeriana officinalis), and zataria (Zataria multiflora)) and five non-herbal supplements (fish oil, melatonin, vitamins B1 and E, and zinc sulphate) in a variety of formulations and doses. Comparators included other supplements, placebo, no treatment, and NSAIDs.We judged all the evidence to be of low or very low quality. The main limitations were imprecision due to very small sample sizes, failure to report study methods, and inconsistency. For most comparisons there was only one included study, and very few studies reported adverse effects. Effectiveness of supplements for primary dysmenorrhoea We have presented pain scores (all on a visual analogue scale (VAS) 0 to 10 point scale) or rates of pain relief, or both, at the first post-treatment follow-up. Supplements versus placebo or no treatmentThere was no evidence of effectiveness for vitamin E (MD 0.00 points, 95% CI -0.34 to 0.34; two RCTs, 135 women).There was no consistent evidence of effectiveness for dill (MD -1.15 points, 95% CI -2.22 to -0.08, one RCT, 46 women), guava (MD 0.59, 95% CI -0.13 to 1.31; one RCT, 151 women); one RCT, 73 women), or fennel (MD -0.34 points, 95% CI -0.74 to 0.06; one RCT, 43 women).There was very limited evidence of effectiveness for fenugreek (MD -1.71 points, 95% CI -2.35 to -1.07; one RCT, 101 women), fish oil (MD 1.11 points, 95% CI 0.45 to 1.77; one RCT, 120 women), fish oil plus vitamin B1 (MD -1.21 points, 95% CI -1.79 to -0.63; one RCT, 120 women), ginger (MD -1.55 points, 95% CI -2.43 to -0.68; three RCTs, 266 women; OR 5.44, 95% CI 1.80 to 16.46; one RCT, 69 women), valerian (MD -0.76 points, 95% CI -1.44 to -0.08; one RCT, 100 women), vitamin B1 alone (MD -2.70 points, 95% CI -3.32 to -2.08; one RCT, 120 women), zataria (OR 6.66, 95% CI 2.66 to 16.72; one RCT, 99 women), and zinc sulphate (MD -0.95 points, 95% CI -1.54 to -0.36; one RCT, 99 women).Data on chamomile and cinnamon versus placebo were unsuitable for analysis. Supplements versus NSAIDSThere was no evidence of any difference between NSAIDs and dill (MD 0.13 points, 95% CI -1.01 to 1.27; one RCT, 47 women), fennel (MD -0.70 points, 95% CI -1.81 to 0.41; one RCT, 59 women), guava (MD 1.19, 95% CI 0.42 to 1.96; one RCT, 155 women), rhubarb (MD -0.20 points, 95% CI -0.44 to 0.04; one RCT, 45 women), or valerian (MD points 0.62 , 95% CI 0.03 to 1.21; one RCT, 99 women),There was no consistent evidence of a difference between Damask rose and NSAIDs (MD -0.15 points, 95% CI -0.55 to 0.25; one RCT, 92 women).There was very limited evidence that chamomile was more effective than NSAIDs (MD -1.42 points, 95% CI -1.69 to -1.15; one RCT, 160 women). Supplements versus other supplementsThere was no evidence of a difference in effectiveness between ginger and zinc sulphate (MD 0.02 points, 95% CI -0.58 to 0.62; one RCT, 101 women). Vitamin B1 may be more effective than fish oil (MD -1.59 points, 95% CI -2.25 to -0.93; one RCT, 120 women). Effectiveness of supplements for secondary dysmenorrhoea There was no strong evidence of benefit for melatonin compared to placebo for dysmenorrhoea secondary to endometriosis (data were unsuitable for analysis). Safety of supplements Only four of the 27 included studies reported adverse effects in both treatment groups. There was no evidence of a difference between the groups but data were too scanty to reach any conclusions about safety. There is no high quality evidence to support the effectiveness of any dietary supplement for dysmenorrhoea, and evidence of safety is lacking. However for several supplements there was some low quality evidence of effectiveness and more research is justified.


The impact of stapling devices use on patients with increased body mass index treated with radical cystectomy.
The aim of this retrospective study was to evaluate the impact of stapling devices use on overweight and obese bladder cancer patients treated with radical cystectomy (RC). Fifty-two overweight and 24 obese patients underwent open RC for muscle invasive bladder cancer. Bladder removal was performed using standard suture technique (SST) or multifire autosuture articulated vascular Endo-GIA (eG). Twenty-three overweight and 11 obese patients formed the SST arms and the remaining 29 overweight and 13 obese patients formed the eG arms. Intra and postoperative parameters and early postoperative complications (30 days) using the Clavien-Dindo classification were recorded. SST and eG arms of overweight and obese patients were comparable in intra and postoperative parameters. Both overweight and obese eG arms had significantly lower estimated blood loss, lower number of transfused packed red blood cells units (PRBC) and lower cystectomy intraoperative time compared with the SST ones (p < 0.05). In obese patients, staplers use led to significantly lower total length of stay compared with SST (p = 0.041). Complications rate was significantly higher in the obese group compared with the overweight group (58.33% versus 30.77%, p = 0.042). No difference in complications was found between the SST and eG arms of the overweight and obese patients. No deaths occurred. Staplers use in RC in overweight/obese patients is accompanied by significantly shorter intraoperative time, lower blood loss and lower number of transfused PRBC units, compared with SST. In obese patients, eG use led to shorter length of stay.


Hydrogen peroxide excretion by oral streptococci and effect of lactoperoxidase-thiocyanate-hydrogen peroxide.
Approved type strains of Streptococcus sanguis, S. mitis, S. mutans, and S. salivarius were grown under aerobic and anaerobic conditions. The rate of hydrogen peroxide excretion, oxygen uptake, and acid production from glucose by washed-cell suspensions of these strains were studied, and the levels of enzymes in cell-free extracts which reduced oxygen, hydrogen peroxide, or hypothiocyanite (OSCN-) in the presence of NADH or NADPH were assayed. The effects of lactoperoxidase-thiocyanate-hydrogen peroxide on the rate of acid production and oxygen uptake by intact cells, the activity of glycolytic enzymes in cell-free extracts, and the levels of intracellular glycolytic intermediates were also studied. All strains consumed oxygen in the presence of glucose. S. sanguis, S. mitis, and anaerobically grown S. mutans excreted hydrogen peroxide. There was higher NADH oxidase and NADH peroxidase activity in aerobically grown cells than in anaerobically grown cells. NADPH oxidase activity was low in all species. Acid production, oxygen uptake, and, consequently, hydrogen peroxide excretion were inhibited in all the strains by lactoperoxidase-thiocyanate-hydrogen peroxide. S. sanguis and S. mitis had a higher capacity than S. mutans and S. salivarius to recover from this inhibition. Higher activity in the former strains of an NADH-OSCN oxidoreductase, which converted OSCN- into thiocyanate, explained this difference. The change in levels of intracellular glycolytic intermediates after inhibition of glycolysis by OSCN- and the actual activity of glycolytic enzymes in cell-free extracts in the presence of OSCN- indicated that the primary target of OSCN- in the glycolytic pathway was glyceraldehyde 3-phosphate dehydrogenase.


[Prosthetic voice rehabilitation after laryngectomy. Failures and complications after previous radiation therapy].
Indwelling voice prostheses are state of the art for post-laryngectomy voice rehabilitation. The aim of this study was to identify the impact of radiation prior to tracheoesophageal puncture on success rate and complications. We undertook a retrospective study of 145 patients who had undergone prosthetic voice restoration between 1990 and 2002 (Provox) and Provox2). Risks of functional failure and complications in 17 patients with previous radiation therapy were compared to those of 128 patients without such therapy. Previous radiation increased not only the risk of functional failure by 2.9 (P=0.023), but also the risk of shunt-related complications such as aspiration around the prosthesis (1.51; P=0.046), widening of the shunt (2.32; P=0.014), esophageal (2.51; P=0.013) or tracheal (3.29; P=0.0023) dislocation of the prosthesis and spontaneous (2.51; P=0.047) or surgical closure (3.76; P=0.037) of the shunt. Primary tracheoesophageal puncture during laryngectomy is recommended in cases without previous radiation therapy, especially when post-laryngectomy radiation is likely. In patients with previous radiation therapy, generally good success rates decrease, however, without absolute contraindication of tracheoesophageal puncture. These results may affect salvage surgery concepts.


[Endoprosthesis of great joints of extremities in difficult anatomic conditions].
Experience in endoprosthesis of great joints in 408 patients in difficult clinical cases associated with defects and deformations of bones of posttraumatic, oncological, dysplastic and systemic character, and also in the cases of complications after primary endoprosthesis is analyzed. Special constructions of implants, instruments, surgical technique were used depending on anatomic and functional changes. Results of endoprosthesis of great joints were accessed as successful in 95% patients in maximal period of follow-up during 13 years.


[On a connection of early manifestations of angle closure glaucoma and development of posterior vitreous detachment].
In author's opinion development of posterior vitreous detachment (PVD) is an involutional factor which participates in pathogenesis of primary angle closure glaucoma (PACG) in predisposed eyes. Initial increase of intraocular pressure in the eyes with narrow anterior chamber angle and predisposition to PACG is closely connected with the moment of PVD development, which can be detected with ultrasonography. The author supposes that ultrasonic examination of the posterior globe to detect PVD can be used as one of monitoring method in patients predisposed to PACG.


Molecular alignment in molecular fluids induced by coupling between density and thermal gradients.
We investigate, using non-equilibrium molecular dynamics simulations and theory, the response of molecular fluids confined in slit pores under the influence of a thermal gradient and/or an applied force. The applied force which has the same functional form as a gravitational force induces an inhomogeneous density in the confined fluid, which results in a net orientation of the molecules with respect to the direction of the force. The orientation is qualitatively similar to that induced by a thermal gradient. We find that the average degree of orientation is proportional to the density gradient of the fluid in the confined region and that the orientation increases with the magnitude of the force. The concurrent application of the external force and the thermal gradient allows us to disentangle the different mechanisms leading to the thermal orientation of molecular fluids. One mechanism is connected to the density variation of the fluid, while the second mechanism can be readily observed in molecular fluids consisting of molecules with mass or size asymmetry, even in the absence of a density gradient, hence it is connected to the application of the thermal gradient only.


Evidence of Anaplasma phagocytophilum and Borrelia burgdorferi infection in cats after exposure to wild-caught adult Ixodes scapularis.
Cats are infected by Anaplasma phagocytophilum and Borrelia burgdorferi when exposed to infected Ixodes scapularis (black-legged ticks). The purpose of our study was to allow wild-caught I. scapularis to feed on healthy research cats (n = 4) and temporally evaluate for A. phagocytophilum DNA in blood by a polymerase chain reaction (PCR) assay as well as for antibody responses to the B. burgdorferi C6 peptide, to the A. phagocytophilum P44 peptide, and to a novel A. phagocytophilum peptide (P44-4). Prior to I. scapularis infestation, all cats were negative for antibodies against both organisms based on a kit optimized for dog serum, and negative for A. phagocytophilum DNA in blood using a conventional PCR assay. Using the pre-infestation samples, an enzyme-linked immunosorbent assay for detecting antibodies against the P44-4 peptide was optimized. Cats were infested with wild-caught I. scapularis for 7 days. Genomic DNA of A. phagocytophilum was amplified from the blood before antibodies were detected in all 4 cats. Antibodies against the C6 peptide, P44 peptide, and P44-4 peptide were detected in the sera of all 4 cats. Antibodies against P44-4 were detected prior to those against P44 in 3 out of 4 cats. The results suggest that a PCR assay should be considered in acutely ill cats with suspected anaplasmosis that are seronegative.


[Determination of 13C abundance in trace amounts of plant metabolites by the infrared 13CO2 analyzer].
The infrared 13CO2 analyzer was improved to measure the 13C abundance in the trace amounts of plant metabolites separated by high performance liquid chromatography (HPLC). To improve the sensitivity to 12C, the range of wave number of 2 360 +/- 10 cm-1 was used for 12C determination. Free carbohydrates in corn leaves fed 13CO2 were separated by HPLC and the 13C abundances in sucrose, glucose and fructose were determined by this improved method. The samples containing 10-40 micrograms of carbon could be successfully analysed.


Effects of Elevated Ozone Levels on Photosynthesis, Biomass and Non-structural Carbohydrates of Phoebe bournei and Phoebe zhennan in Subtropical China.
To assess the impacts of ozone (O3) on carbon metabolism of subtropical broadleaved tree species, seedlings of Phoebe bournei and Phoebe zhennan were exposed to elevated O3 levels in open-top chambers (OTCs) from June to November 2014. Three treatments were conducted in nine total OTCs, including charcoal-filter air (CF) as a control treatment, low O3 treatment 'O3-1' (∼100 nl l-1), and high O3 treatment 'O3-2' (∼150 nl l-1). Our findings demonstrated that elevated O3 levels significantly decreased the net photosynthesis rates (Pn ) and leaf, root, and total biomass of both species, while it did not significantly affect the root/shoot ratio in P. bournei and P. zhennan. O3-1 treatments significantly increased water soluble carbohydrates (WSC) in leaves of both tree species, while only increased the total non-structural carbohydrates (TNC) and starch in leaves of P. bournei; effects on P. zhennan were equivalent in comparison to the control treatment (CF). Likewise, there was no effect of treatment on the polysaccharide content of both tree species. The contents of polysaccharide, starch contents in fine roots of both species, and TNC in fine roots of P. bournei increased significantly in O3-1 compared to CF. O3-2 treatment significantly decreased starch and TNC in the fine roots of P. bournei, and significantly decreased polysaccharide, starch, WSC, and TNC in the fine roots of P. zhennan. Elevated O3 had no effects on leaf polysaccharide in both species, but O3-1 significantly increased polysaccharide in the fine roots of both species, and O3-1 significantly increased WSC in the leaves while decreased that in the fine roots of both species. These results suggested that elevated O3 levels have significant impacts on the carbon metabolism of both tree species in our study, with differential responses between tree species and among leaves and roots.


Efficient sequence analysis of the six gene products (7-74 kDa) from the Escherichia coli thiamin biosynthetic operon by tandem high-resolution mass spectrometry.
The 10(5) resolving power and MS/MS capabilities of Fourier-transform mass spectrometry provide electrospray ionization mass spectra containing >100 molecular and fragment ion mass values of high accuracy. Applying these spectra to the detection and localization of errors and modifications in the DNA-derived sequences of proteins is illustrated with the thiCEFSGH thiamin biosynthesis operon from Escherichia coli. Direct fragmentation of the multiply-charged intact protein ions produces large fragment ions covering the entire sequence; further dissociation of these fragment ions provides information on their sequences. For ThiE (23 kDa), the entire sequence was verified in a single spectrum with an accurate (0.3 Da) molecular weight (Mr) value, with confirmation from MS/MS fragment masses. Those for ThiH (46 kDa) showed that the Mr value (1 Da error) represented the protein without the start Met residue. For ThiF (27 kDa), MS/MS localized a sequence discrepancy to a 34 residue peptide. The first 107 residues of ThiC (74 kDa) were shown to be correct, with C-terminal heterogeneity indicated. For ThiG (predicted Mr = 34 kDa), ESI/FTMS showed two components of 7,310.74 (ThiS) and 26,896.5 Da (ThiG); MS/MS uncovered three reading frame errors and a stop codon for the first protein. MS/MS ions are consistent with 68 fragments predicted by the corrected ThiS/ThiG DNA sequences.