Age differences in brain activity during perceptual versus reflective attention.
This functional magnetic resonance imaging study presented participants with a face and scene simultaneously on each trial, and assessed the impact of perceptual versus reflective selective attention on activity in parahippocampal place area. Young and older adults showed equivalent activation in parahippocampal place area when cued to attend to the scene when the stimuli were perceptually present and when cued to refresh (briefly think about) the scene after the stimuli were no longer present. The groups also showed equivalent deactivation when cued to attend to the face when the stimuli were perceptually present. However, older adults showed less deactivation than young adults when cued to refresh the face, providing evidence for greater age-related disruption of reflective than perceptual selective attention.


Neural cell fate in rca1 and cycA mutants: the roles of intrinsic and extrinsic factors in asymmetric division in the Drosophila central nervous system.
In the central nervous system (CNS) of Drosophila embryos lacking regulator of cyclin A (rca1) or cyclin A, we observe that several ganglion mother cells (GMCs) fail to divide. Whereas GMCs normally produce two sibling neurons that acquire different fates ('A/B'), non-dividing GMCs differentiate exclusively in the manner of one of their progeny ('B'). In zygotic numb mutants, sibling neuron fate alterations ('A/B' to 'A/A') occur infrequently or do not occur in some sibling pairs; we have determined that depletion of both maternal and zygotic numb causes sibling neurons to acquire equalized fates ('A/A') with near-complete expressivity. In rca1, numb mutant embryos, we observe binary cell fate changes ('B' to 'A') in several GMCs as well. Finally, we have demonstrated that expression of Delta in the mesoderm is sufficient to attain both sibling fates. Our results indicate that the intrinsic determinant Numb is absolutely required to attain differential sibling neuron fates. While the extrinsic factors Notch and Delta are also required to attain both fates, our results indicate that Delta signal can be received from outside the sibling pair.


[Analysis of dietary factors in Alzheimer's disease: clinical use of nutritional intervention for prevention and treatment of dementia].
To determine dietary factors involved in the pathological process of Alzheimer's disease (AD), we analyzed food consumption and intake of nutrients using Self-administered Diet History Questionnaire (DHQ) developed for Japanese. Sixty four AD patients and 80 age-matched healthy subjects were enrolled in this study. AD was diagnosed according to the criteria of DSM-IV. Dietary behaviors of AD patients was markedly deviated from those of age-matched healthy elderly. AD patients disliked fish and green-yellow vegetables and took more meats than controls. Energy-adjusted analysis of nutrients revealed that AD patients took less vitamin C and carotene. Most conspicuously, AD patients took significantly smaller amount of n-3 polyunsaturated fatty acid (PUFA) reflecting low consumption of fish, and their n-6/n-3 ratio was significantly increased. These habits started from 3 months to 44 years before the onset of dementia, suggesting these dietary abnormalities are not merely the consequence of dementia. Rather, it implies that AD might be a life style-related disease such as coronary heart disease, western style diet-associated cancer and hyperallergy. To see if cognitive function was improved by correcting the n-6/n-3 ratio, we prescribed eicossapentaenoic acid (EPA), one type of n-3 PUFA, for AD patients. Cognitive function was evaluated using MMSE. Administration of EPA (900 mg/day) improved MMSE significantly with maximal effects at 3 months and the effects lasted 6 months. However, the score of MMSE decreased after 6 months. The present study showed that nutritional intervention is useful for the prevention of AD, and also for the therapy of dementia, though it has some limitation.


Inter-rater and intra-rater reliability in the interpretation of MTI Photoscreener photographs of Native American preschool children.
To evaluate inter- and intra-rater reliability for the interpretation of MTI Photoscreener photographs taken in a population of Native American preschool children with a high prevalence of astigmatism. Photographs of 369 children were rated by 11 nonexpert and 3 expert raters. Photographs for each child were scored as pass, refer, or retake. Nonexpert raters scored photos on two separate occasions, permitting analysis of intra-rater reliability. Analyses of pass/refer responses only: inter-rater reliability was moderate to substantial among nonexpert raters and substantial among expert raters. Intra-rater reliability among nonexperts was substantial. Analyses of all responses (pass, refer, and retake): inter-rater reliability for pass and refer scores was moderate among nonexperts and substantial among experts; for retake scores inter-rater reliability was slight for nonexperts and moderate for experts. Intra-rater reliability among nonexperts was substantial for pass and refer scores and moderate for retake scores. In this population with a high prevalence of astigmatism, whether MTI photoscreening results are interpretable is much more variable among and within raters than whether an interpretable photograph should be scored as pass or refer. The level of agreement among raters in the current study was influenced by the experience of the raters. In addition, nonexpert raters were more likely to deem a photograph uninterpretable than expert raters.


Per- and Polyfluoroalkyl Substances (PFAS) in Breast Milk: Concerning Trends for Current-Use PFAS.
This is the first study in the last 15 years to analyze per- and polyfluoroalkyl substances (PFAS) in breast milk collected from mothers (n = 50) in the United States, and our findings indicate that both legacy and current-use PFAS now contaminate breast milk, exposing nursing infants. Breast milk was analyzed for 39 PFAS, including 9 short-chain and 30 long-chain compounds, and 16 of these PFAS were detected in 4-100% of the samples. The ∑PFAS concentration in breast milk ranged from 52.0 to 1850 pg/mL with a median concentration of 121 pg/mL. Perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) were the most abundant PFAS in these samples (medians 30.4 and 13.9 pg/mL, respectively). Two short-chain PFAS, including perfluoro-n-hexanoic acid (PFHxA, C6) and perfluoro-n-heptanoic acid (PFHpA, C7), were detected in most of the samples with median concentrations of 9.69 and 6.10 pg/mL, respectively. Analysis of the available breast milk PFAS data from around the world over the period of 1996-2019 showed that while the levels of the phased-out PFOS and PFOA have been declining with halving times of 8.1 and 17 years, respectively, the detection frequencies of current-use short-chain PFAS have been increasing with a doubling time of 4.1 years.


[Antioxidant activity of mildronate and L-carnitine in the treatment of patients with cerebrovascular diseases].
The antioxidant properties of mildronate and a structurally close compound L-carnitine were studied under clinical conditions during the therapy of patients with acute lacunar stroke and discirculatory encephalopathy (DEP) on the background of diabetes mellitus, respectively. Administered in addition to the base course of therapy, both mildronate (in a daily dose of 500 mg) and L-carnitine (2 mg) increased the resistance of blood serum lipoproteins with respect to peroxidation. It was concluded that the drugs possess antioxidant activity and offer protection against lipid peroxidation. L-carnitine acute produced a significant hypoglycemic action and made possible an almost twofold (42%) decrease in the dose of hypoglycemic drugs. The administration of L-carnitine also improved both abstract and concrete thinking and memory function in DEP patients. The results allowed mildronate and L-carnitine to be included in the complex therapy of patients with cerebrovascular diseases.


An integrated Bayesian analysis of LOH and copy number data.
Cancer and other disorders are due to genomic lesions. SNP-microarrays are able to measure simultaneously both genotype and copy number (CN) at several Single Nucleotide Polymorphisms (SNPs) along the genome. CN is defined as the number of DNA copies, and the normal is two, since we have two copies of each chromosome. The genotype of a SNP is the status given by the nucleotides (alleles) which are present on the two copies of DNA. It is defined homozygous or heterozygous if the two alleles are the same or if they differ, respectively. Loss of heterozygosity (LOH) is the loss of the heterozygous status due to genomic events. Combining CN and LOH data, it is possible to better identify different types of genomic aberrations. For example, a long sequence of homozygous SNPs might be caused by either the physical loss of one copy or a uniparental disomy event (UPD), i.e. each SNP has two identical nucleotides both derived from only one parent. In this situation, the knowledge of the CN can help in distinguishing between these two events. To better identify genomic aberrations, we propose a method (called gBPCR) which infers the type of aberration occurred, taking into account all the possible influence in the microarray detection of the homozygosity status of the SNPs, resulting from an altered CN level. Namely, we model the distributions of the detected genotype, given a specific genomic alteration and we estimate the parameters involved on public reference datasets. The estimation is performed similarly to the modified Bayesian Piecewise Constant Regression, but with improved estimators for the detection of the breakpoints.Using artificial and real data, we evaluate the quality of the estimation of gBPCR and we also show that it outperforms other well-known methods for LOH estimation. We propose a method (gBPCR) for the estimation of both LOH and CN aberrations, improving their estimation by integrating both types of data and accounting for their relationships. Moreover, gBPCR performed very well in comparison with other methods for LOH estimation and the estimated CN lesions on real data have been validated with another technique.


Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp. by southern hybridization with opd from Pseudomonas diminuta.
Parathion hydrolases have been previously described for an American isolate of Pseudomonas diminuta and a Philippine isolate of Flavobacterium sp. (ATCC 27551). The gene which encodes the broad-spectrum organophosphate phosphotriesterase in P. diminuta has been shown by other investigators to be located on a 66-kilobase (kb) plasmid. The intact gene (opd, organophosphate-degrading gene) from this degradative plasmid was cloned into M13mp10 and found to express parathion hydrolase under control of the lac promoter in Escherichia coli. In Flavobacterium sp. strain ATCC 27551, a 43-kb plasmid was associated with the production of parathion hydrolase by curing experiments. The M13mp10-cloned fragment of the opd gene from P. diminuta was used to identify a homologous genetic region from Flavobacterium sp. strain ATCC 27551. Southern hybridization experiments demonstrated that a genetic region from the 43-kb Flavobacterium sp. plasmid possessed significant homology to the opd sequence. Similar hybridization did not occur with three other native Flavobacterium sp. plasmids (approximately 23, 27, and 51 kb) present within this strain or with genomic DNA from cured strains. Restriction mapping of various recombinant DNA molecules containing subcloned fragments of both opd plasmids revealed that the restriction maps of the two opd regions were similar, if not identical, for all restriction endonucleases tested thus far. In contrast, the restriction maps of the cloned plasmid sequences outside the opd regions were not similar. Thus, it appears that the two discrete bacterial plasmids from parathion-hydrolyzing soil bacteria possess a common but limited region of sequence homology within potentially nonhomologous plasmid structures.


The visual stresses of the aerospace environment.
The aviator or astronaut is subject to alterations of the environment which may seriously affect his performance or survivability. In a series of experiments, the stresses of hypoxia, vibration, high and low gravity, heat and noise were investigated to determine their effects upon visual function. With the possible exceptions of the latter two, all were found to produce measurable decrements in performance even at moderate levels; however, it is most remarkable that significant capability remains even in rather severe stress. Certain hypotheses regarding these visual decrements that have appeared in the literature are rejected on the basis of experimental evidence.


Modifiable Midlife Risk Factors for Late-Life Cognitive Impairment and Dementia.
The baby boom generation is approaching the age of greatest risk for cognitive impairment and dementia. There is growing interest in strategies to modify the environment in midlife to increase the probability of maintaining cognitive health in late life. Several potentially modifiable risk factors have been studied in relation to cognitive impairment and dementia in late life, but methodological limitations of observational research have resulted in some inconsistencies across studies. The most promising strategies are maintaining cardiovascular health, engagement in mental, physical, and social activities, using alcohol in moderation, abstaining from tobacco use, and following a heart-healthy diet. Other factors that may influence cognitive health are occupational attainment, depression, personality, exposure to general anesthesia, head injury, postmenopausal hormone therapy, non-steroidal anti-inflammatory medications, and nutritional supplements such as antioxidants. Some long-term observational studies initiated in midlife or earlier, and some randomized controlled trials, have examined the effects of specific cognitive health promotion behaviors in midlife on the risk of cognitive impairment in late life. Overall, these studies provide limited support for risk reduction at this time. Recommendations and challenges for developing effective strategies to reduce the burden of cognitive impairment and dementia in the future are discussed.


Tinnitus: patients do not have to 'just live with it'.
Tinnitus is distressing and affects the quality of life for many patients. Because primary care physicians may be the entry point for patients seeking help for tinnitus, we urge them to acknowledge this symptom and its potential negative impact on the patient's health and quality of life. Physicians should actively listen to the patient and provide hope and encouragement, but also provide realistic expectations about the course of treatment. The patient must also understand that there may be no singular "cure" for tinnitus and that management may involve multidisciplinary assessment and treatment.


T lymphocytes targeting native receptors.
The adoptive transfer of T cells specific for native tumor antigens (TAs) is an increasingly popular cancer treatment option because of the ability of these cells to discriminate between normal and tumor tissues and the corresponding lack of short or long-term toxicities. Infusions of antigen-specific CD4(+) and CD8(+) T cells targeting viral antigens derived from Epstein-Barr virus (EBV) induce sustained complete tumor remissions in patients with highly immunogenic tumors such as post-transplant lymphoproliferative disease, although resistance occurred when the infused T-cell population had restricted antigen specificity. T cells specific for EBV antigens have also produced complete remissions of EBV-positive nasopharyngeal carcinomas and lymphomas developing in immunocompetent individuals, even though in these patients tumor survival is dependent on their ability to evade T-cell immunity. Adapting this strategy to non-viral tumors is more challenging, as the target antigens expressed are less immunogenic and the tumors lack the potent danger signals that are characteristic of viruses. The goals of current studies are to define conditions that promote expansion of antigen-specific T cells ex vivo and to ensure their in vivo persistence and survival by combining with maneuvers such as lymphodepletion, checkpoint inhibition, cytokine infusions, or genetic manipulations. More pragmatic goals are to streamline manufacturing to facilitate the transition of these therapies to late phase trials and to evaluate closely histocompatibility antigen (HLA)-matched banked antigen-specific T cells so that T-cell therapies can be made more broadly available.


Synaptophysin: leading actor or walk-on role in synaptic vesicle exocytosis?
Synaptophysin (Syp) was the first synaptic vesicle (SV) protein to be cloned. Since its discovery in 1985, it has been used by us and by many laboratories around the world as an invaluable marker to study the distribution of synapses in the brain and to uncover the basic features of the life cycle of SVs. Although single gene ablation of Syp does not lead to an overt phenotype, a large body of experimental data both in vitro and in vivo indicate that Syp (alone or in association with homologous proteins) is involved in multiple, important aspects of SV exo-endocytosis, including regulation of SNARE assembly into the fusion core complex, formation of the fusion pore initiating neurotransmitter release, activation of SV endocytosis and SV biogenesis. In this article, we summarise the main results of the studies on Syp carried out by our and other laboratories, and explain why we believe that Syp plays a major role in SV trafficking.


On the regularities of distribution of Hypoderma bovis De Geer larvae parasitizine cattle herds in different parts of the range of this warble fly.
The type and parameters of the distribution of the second and third instar larvae of Hypoderma bovis in cattle herds in Czechoslovakia (54 herds, 7233 head), Mongolia (20 herds, 1809 head) and the USSR (48 herds, 4978 head) were studied. A statistical analysis showed a) that in the majority of cases negative binomial distribution serves as a model of the distribution of larvae with sufficient reliability; b) that the regularity of dependence of the distribution exponent k of the negative binomial distribution on the incidence of infestation remains constant in different parts of the range of thes warble fly. The latter fact indicates that the regulatory systems limiting the population numbers of this warble fly are associated with host-parasite relationships and do not depend on a complex of conditions specific for various natural zones. In order to understand the regulatory processes in parasite populations it is necessary to study equally the regulatory mechanisms operating primarily on the level of specimens as well as the regulatory systems operating on the level of populations.


The COBRA II (COmplex Bifurcation lesions: RAndomized comparison of a fully bioresorbable modified-T stenting strategy versus bifurcation reconstruction with a dedicated self-expanding stent in combination with bioresorbable scaffolds) study: Final 5-year follow-up.
In order to advocate further research in bioresorbable scaffold (BVS) technology we report the final 5-year outcomes of the COBRA II study, the only randomized controlled trial (RCT) performed to investigate the safety, feasibility, and performance of Absorb BVS (Abbott Vascular) in true coronary bifurcations. COBRA II was a prospective single-center RCT. Fifteen patients with true coronary bifurcation lesions were randomized to bifurcation treatment with self-expanding biolimus-eluting Axxess bifurcation device (Biosensors International) combined with additional bioresorbable everolimus-eluting Absorb BVS in the bifurcation branches on (Axxess group) or to 2-stent mod-T stenting technique with Absorb BVS (mod-T group). Optical coherence tomography was performed post-procedure and at 30-months. The primary endpoint was change in minimal luminal area (MLA) on OCT from baseline to 30-months. Fifteen patients with coronary bifurcation lesions were randomized to the Axxess group (n = 8) or Modified-T group (n = 7). At 30 months, MLAs were significantly smaller than post-procedure in the majority of bifurcation segments treated with BVS due to neointima formation, while MLAs in the proximal Axxess segment remained stable (primary endpoint). Five-year clinical follow-up was available for all patients. Only 1 major adverse cardiac event occurred; a patient underwent target lesion revascularization at 30 months in the Axxess group. There were no cases of cardiac death, spontaneous MI, or stent/scaffold thrombosis. In this small RCT bifurcation study, BVS luminal dimensions were significantly smaller at 30 months, with acute strut discontinuities and late Intraluminal dismantling frequently observed, although acceptable clinical outcomes were noted at 5 years.


Diagnosis of gestational diabetes mellitus: falling through the net.
Gestational diabetes mellitus (GDM) is associated with increased risks to mother and child, but globally agreed diagnostic criteria remain elusive. Identification of women with GDM is important, as treatment reduces adverse outcomes such as perinatal death, shoulder dystocia and neonatal hypoglycaemia. Recently, the UK's National Institute for Health and Care Excellence (NICE) recommended new diagnostic thresholds for GDM which are different from the International Association of the Diabetes and Pregnancy Study Groups (IADPSG) criteria endorsed by the WHO. The study aim was to assess neonatal and obstetric outcomes among women who would test positive for the IADPSG criteria but negative for the NICE 2015 criteria. Data from 25,543 consecutive singleton live births (2004-2008) were obtained retrospectively from hospital records. Women were screened with a random plasma glucose (RPG; 12-16 weeks) and a 50 g glucose challenge test (GCT; 26-28 weeks). If RPG >7.0 mmol/l, GCT >7.7 mmol/l or symptoms were present, a 75 g OGTT was offered (n = 3,848). In this study, GDM prevalence was 4.13% (NICE 2015) and 4.62% (IADPSG). Women who 'fell through the net', testing NICE-negative but IADPSG-positive (n = 387), had a higher risk of having a large-for-gestational-age (LGA) infant (birthweight >90th percentile for gestational age; adjusted OR [95% CI] 3.12 [2.44, 3.98]), Caesarean delivery (1.44 [1.15, 1.81]) and polyhydramnios (6.90 [3.94, 12.08]) compared with women with negative screening results and no OGTT (n = 21,695). LGA risk was highest among women with fasting plasma glucose 5.1-5.5 mmol/l (n = 167): the mean birthweight was 350 g above that of the reference population and 37.7% of infants were LGA. The IADPSG criteria identify women at substantial risk of complications who would not be identified by the NICE 2015 criteria.


A History of Drug Discovery for Treatment of Nausea and Vomiting and the Implications for Future Research.
The origins of the major classes of current anti-emetics are examined. Serendipity is a recurrent theme in discovery of their anti-emetic properties and repurposing from one indication to another is a continuing trend. Notably, the discoveries have occurred against a background of company mergers and changing anti-emetic requirements. Major drug classes include: (i) Muscarinic receptor antagonists-originated from historical accounts of plant extracts containing atropine and hyoscine with development stimulated by the need to prevent sea-sickness among soldiers during beach landings; (ii) Histamine receptor antagonists-searching for replacements for the anti-malaria drug quinine, in short supply because of wartime shipping blockade, facilitated the discovery of histamine (H1) antagonists (e.g., dimenhydrinate), followed by serendipitous discovery of anti-emetic activity against motion sickness in a patient undergoing treatment for urticaria; (iii) Phenothiazines and dopamine receptor antagonists-investigations of their pharmacology as "sedatives" (e.g., chlorpromazine) implicated dopamine receptors in emesis, leading to development of selective dopamine (D2) receptor antagonists (e.g., domperidone with poor ability to penetrate the blood-brain barrier) as anti-emetics in chemotherapy and surgery; (iv) Metoclopramide and selective 5-hydroxytryptamine3(5-HT3) receptor antagonists-metoclopramide was initially assumed to act only via D2 receptor antagonism but subsequently its gastric motility stimulant effect (proposed to contribute to the anti-emetic action) was shown to be due to 5-hydroxytryptamine4 receptor agonism. Pre-clinical studies showed that anti-emetic efficacy against the newly-introduced, highly emetic, chemotherapeutic agent cisplatin was due to antagonism at 5-HT3 receptors. The latter led to identification of selective 5-HT3 receptor antagonists (e.g., granisetron), a major breakthrough in treatment of chemotherapy-induced emesis; (v) Neurokinin1receptor antagonists-antagonists of the actions of substance P were developed as analgesics but pre-clinical studies identified broad-spectrum anti-emetic effects; clinical studies showed particular efficacy in the delayed phase of chemotherapy-induced emesis. Finally, the repurposing of different drugs for treatment of nausea and vomiting is examined, particularly during palliative care, and also the challenges in identifying novel anti-emetic drugs, particularly for treatment of nausea as compared to vomiting. We consider the lessons from the past for the future and ask why there has not been a major breakthrough in the last 20 years.


Galectin-13/placental protein 13: redox-active disulfides as switches for regulating structure, function and cellular distribution.
Galectin-13 (Gal-13) plays numerous roles in regulating the relationship between maternal and fetal tissues. Low expression levels or mutations of the lectin can result in pre-eclampsia. The previous crystal structure and gel filtration data show that Gal-13 dimerizes via formation of two disulfide bonds formed by Cys136 and Cys138. In the present study, we mutated them to serine (C136S, C138S and C136S/C138S), crystalized the variants and solved their crystal structures. All variants crystallized as monomers. In the C136S structure, Cys138 formed a disulfide bond with Cys19, indicating that Cys19 is important for regulation of reversible disulfide bond formation in this lectin. Hemagglutination assays demonstrated that all variants are inactive at inducing erythrocyte agglutination, even though gel filtration profiles indicate that C136S and C138S could still form dimers, suggesting that these dimers do not exhibit the same activity as wild-type (WT) Gal-13. In HeLa cells, the three variants were found to be distributed the same as with WT Gal-13. However, a Gal-13 variant (delT221) truncated at T221 could not be transported into the nucleus, possibly explaining why women having this variant get pre-eclampsia. Considering the normally high concentration of glutathione in cells, WT Gal-13 should exist mostly as a monomer in cytoplasm, consistent with the monomeric variant C136S/C138S, which has a similar ability to interact with HOXA1 as WT Gal-13.


The effect of skin-to-skin contact (kangaroo care) shortly after birth on the neurobehavioral responses of the term newborn: a randomized, controlled trial.
The method of skin-to-skin contact (kangaroo care [KC]) has shown physiologic, cognitive, and emotional gains for preterm infants; however, KC has not been studied adequately in term newborns. To evaluate the effect of KC, used shortly after delivery, on the neurobehavioral responses of the healthy newborn. A randomized, controlled trial using a table of random numbers. After consent, the mothers were assigned to 1 of 2 groups: KC shortly after delivery or a no-treatment standard care (control group). Included were 47 healthy mother-infant pairs. KC began at 15 to 20 minutes after delivery and lasted for 1 hour. Control infants and KC infants were brought to the nursery 15 to 20 and 75 to 80 minutes after birth, respectively. During a 1-hour-long observation, starting at 4 hours postnatally, the KC infants slept longer, were mostly in a quiet sleep state, exhibited more flexor movements and postures, and showed less extensor movements. KC seems to influence state organization and motor system modulation of the newborn infant shortly after delivery. The significance of our findings for supportive transition from the womb to the extrauterine environment is discussed. Medical and nursing staff may be well advised to provide this kind of care shortly after birth.


Dysbiosis Modulates Ocular Surface Inflammatory Response to Liposaccharide.
The purpose of this study was to investigate the inflammatory response of cornea and conjunctiva to topically applied lipopolysaccharide (LPS) in mice with and without antibiotic (antibiotic cocktail, ABX) induced dysbiosis. Dysbiosis was induced by oral antibiotics for 14 days in a group of conventional female C57BL/6J (B6) mice. 16S rRNA sequencing investigated microbiome composition. Intestinal microbiome differences were assessed using 16S rRNA sequencing of fecal pellet DNA. Blood was collected after euthanasia. CD86 expression in draining nodes was examined by flow cytometry. At day 15, a single dose of LPS or vehicle was topically applied to ABX and naïve mice. Corneal epithelium and conjunctiva were obtained after 4 hours and processed for gene expression analysis. A separate group of germ-free (GF) B6 mice was also topically challenged with LPS. Antibiotic treatment significantly decreased intestinal diversity and increased serum levels of LPS. This was accompanied by a significant increase in CD86+MHC II+CD11c+CD11b+ cells in draining nodes. Compared to vehicle, topically applied LPS increased IL-1β, TNF-α, and CXCL10 mRNA transcripts in cornea and IL-1β, TNF-α, and CXCL10 in the conjunctiva in conventional and antibiotic-treated groups. However, there was higher TNF-α, CXCL10, and IL-12 expression in the cornea of LPS-treated ABX mice compared to LPS-treated mice with intact microbiota. LPS stimulation on GF conjunctiva mirrored the results in ABX mice, although greater IL-12 and IFN-γ expression was observed in GF conjunctiva compared to conventional LPS-treated mice. Acute depletion of commensals through antibiotics or germ-free environment worsens the inflammatory response to LPS.


Genetic design of biologically inspired receptive fields for neural pattern recognition.
This paper proposes a new method for the design, through simulated evolution, of biologically inspired receptive fields in feedforward neural networks (NNs). The method is intended to enhance pattern recognition performance by creating new neural architectures specifically tuned for a particular pattern recognition problem. It proposes a combined neural architecture composed of two networks in cascade: a feature extraction network (FEN) followed by a neural classifier. The FEN is composed of several layers with receptive fields constructed by additive superposition of excitatory and inhibitory fields. A genetic algorithm (GA) is used to select receptive field parameters to improve classification performance. The parameters are receptive field size, orientation, and bias as well as the number of different receptive fields in each layer. Based on a random initial population where each individual represents a different neural architecture, the GA creates new enhanced individuals. The method is applied to handwritten digit classification and face recognition. In both problems, results show strong dependency between NN classification performance and receptive field architecture. GA selected parameters of the receptive fields produced improvements in the classification performance on the test set up to 90.8% for the problem of handwritten digit classification and up to 84.2% for the face recognition problem. On the same test sets, results were compared advantageously to standard feedforward multilayer perceptron (MLP) NNs where receptive fields are not explicitly defined. The MLP reached a maximum classification performance of 84.9% and 77.5% in both problems, respectively.


Site-specific phosphorylation of the middle molecular weight human neurofilament protein in transfected non-neuronal cells.
We expressed the human midsized neurofilament subunit (NF-M) using genomic DNA in mouse L cells and showed that it is transcribed and translated into a protein capable of assembly into the cytoskeleton and of forming a filamentous network that colocalizes with the endogenous vimentin filaments. Moreover, human NF-M expressed in L cells is phosphorylated at sites within the multiphosphorylation repeat (MPR), i.e., the major sites of phosphorylation of NF-M in vivo. We also expressed a genomic construct lacking the MPR domain in the native molecule and showed that this MPR(-) protein also was expressed and formed a filamentous network despite diminished incorporation of radiolabeled phosphate. Two major conclusions emerged from the work described in this paper: human NF-M is translated, assembled, and phosphorylated at physiological sites without the need of any other specific neuronal proteins; phosphorylation sites other than the MPR are present within NF-M which may play a role in synthesis, assembly, and degradation of NF protein in humans.


Triptolide Decreases Cell Proliferation and Induces Cell Death in Triple Negative MDA-MB-231 Breast Cancer Cells.
Triple negative breast cancers (TNBCs) do not respond to conventional estrogen receptor/progesterone receptor/human epidermal growth factor receptor-2 targeted interventions due to the absence of the respective receptor targets. They are aggressive, exhibit early recurrence, metastasize, are more invasive in nature, and develop drug resistance. Some plant-derived substances have been screened and have gained attention as efficient anticancer drugs for TNBCs with few adverse effects. Here, we evaluate triptolide (concentrations in the range of 100 pM to 10 µM), a di-terpene tri-epoxide isolated from thunder god vine for its efficacy as anticancer drug in MDA-MB-231 TNBC cells. Cell proliferation and viability were assessed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay and trypan blue exclusion assay, respectively. A flow cytometry-based apoptosis assay was performed by using fluorescein isothiocyanate (FITC)-conjugated annexin V and propidium iodide (PI). Western blotting was performed to determine the levels of apoptotic and autophagy proteins such as caspase 3, LC3B and SQSTM1/p62. Results indicate that in 72 h of 1 nM triptolide treatment, the percentage of cell proliferation in MDA-MB-231 cells declined to 49 ± 18.9% (mean ± standard deviation (SD)), whereas the proliferation rate did not drop below 80% in MCF-7 cells (non-TNBC cells which express the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2) even at the highest concentration tested (10 µM). The MDA-MB-468 cells showed a similar trend to MDA-MB-231 cells. Furthermore, triptolide treatment for 72 h significantly decreased cell viability at concentrations above 10 nM. Apoptotic cell death assay in 72 h triptolide-treated MDA-MB-231 cells revealed 29.3 ± 10.57% of early apoptotic cells in comparison to the control group (4.61 ± 2.24%). Cell cycle analysis indicated accumulation of cells in sub G₀/G₁ phase, indicating apoptosis. Western blot analysis in the 25 nM triptolide treatment group revealed induction of autophagy as shown by a significant decrease in the levels of autophagy marker p62 (by 0.2-fold p < 0.0001) and with an increase in the levels of LC3B-II (by 8-fold p < 0.05). An increase in the levels of the apoptotic marker cleaved caspase 3 (by 4-fold p < 0.05) was also observed in triptolide treated MDA-MB-231 cells. Our data shows that triptolide could be an efficient anticancer agent in the treatment of TNBCs.


Investigation of the Sensitivity of Transmission Raman Spectroscopy for Polymorph Detection in Pharmaceutical Tablets.
Polymorph detection is critical for ensuring pharmaceutical product quality in drug substances exhibiting polymorphism. Conventional analytical techniques such as X-ray powder diffraction and solid-state nuclear magnetic resonance are utilized primarily for characterizing the presence and identity of specific polymorphs in a sample. These techniques have encountered challenges in analyzing the constitution of polymorphs in the presence of other components commonly found in pharmaceutical dosage forms. Laborious sample preparation procedures are usually required to achieve satisfactory data interpretability. There is a need for alternative techniques capable of probing pharmaceutical dosage forms rapidly and nondestructively, which is dictated by the practical requirements of applications such as quality monitoring on production lines or when quantifying product shelf lifetime. The sensitivity of transmission Raman spectroscopy for detecting polymorphs in final tablet cores was investigated in this work. Carbamazepine was chosen as a model drug, polymorph form III is the commercial form, whereas form I is an undesired polymorph that requires effective detection. The concentration of form I in a direct compression tablet formulation containing 20% w/w of carbamazepine, 74.00% w/w of fillers (mannitol and microcrystalline cellulose), and 6% w/w of croscarmellose sodium, silicon dioxide, and magnesium stearate was estimated using transmission Raman spectroscopy. Quantitative models were generated and optimized using multivariate regression and data preprocessing. Prediction uncertainty was estimated for each validation sample by accounting for all the main variables contributing to the prediction. Multivariate detection limits were calculated based on statistical hypothesis testing. The transmission Raman spectroscopic model had an absolute prediction error of 0.241% w/w for the independent validation set. The method detection limit was estimated at 1.31% w/w. The results demonstrated that transmission Raman spectroscopy is a sensitive tool for polymorphs detection in pharmaceutical tablets.


A phase II study of gemcitabine in patients with transitional cell carcinoma of the urinary tract previously treated with platinum. Italian Co-operative Group on Bladder Cancer.
The aim of this study was to evaluate the efficacy and safety of gemcitabine, a pyrimidine antimetabolite, in the treatment of advanced transitional cell carcinoma of the urinary tract. 35 patients with unresectable or metastatic transitional cell carcinoma of the urinary tract previously treated with a platinum-based regimen were studied. Gemcitabine was administered at a dosage of 1200 mg/m2 as a 30-min intravenous infusion on days 1, 8 and 15, repeated every 28 days. 31 patients were evaluable for efficacy. 4 patients achieved a complete response (12.9%), 3 a partial response (9.6%) and 13 (42%) were stable for at least 4 weeks (overall response 22.5%; 95% confidence interval 8-37%). The median response duration was 11.8 months (range 3.6-17.7 + months) and median survival for all patients entered was 5 months (range 2-21 + months). 2 patients with complete response are still alive with no evidence of disease after 14 and 21 months. Gemcitabine also provided subjective symptomatic relief from pain, cystitis, dysuria, haematuria and peripheral oedema. Patients experienced little WHO grade 3-4 toxicity, with anaemia in 8 patients (23%), thrombocytopenia in 5 (14.2%), leucopenia in 4 (11.4%) and neutropenia in 7 (20%). WHO grade 3-4 hepatic toxicity occurred in 4 patients (11.4%) and transient elevations of transaminase was noted in 3 (8.6%). No patient had WHO grade 3-4 elevation of serum creatinine level. There was no WHO grade 4 symptomatic toxicity and no alopecia was noted. Transient influenza symptoms with gemcitabine occurred in 18 patients (51.4%) with 13 patients (37.1%) experiencing fever (2.9% WHO grade 3). In conclusion, gemcitabine is an new active agent for the treatment of transitional cell carcinoma of the urinary bladder with a mild toxicity profile; it warrants further investigation in combination with cisplatin in chemotherapy naive patients.


The dynamic still-face effect: do infants decrease bidding over time when parents are not responsive?
The still-face paradigm (SFP) was designed to assess infant expectations that parents will respond to infant communicative signals. During the still-face (SF) episode, the parent ceases interaction and maintains a neutral expression. Original, qualitative descriptions of infant behavior suggested changes within the SF episode: infants decrease bidding and disengage from their impassive parent. Research has documented changes in mean levels of infant behavior between episodes of the SFP. The hypothesis that infant behavior changes within the SF episode has not been empirically tested. In this study, hierarchical linear modeling indicated that infant gazing at the parent, smiling, and social bidding (smiling while gazing at the parent) decreased with time in the SF episode, while infant cry-face expressions increased. Changes in infant behaviors within the SF episode were associated with infant attachment and infant internalizing problems. The dynamic still-face effect quantifies infant initiation of interaction in the face of parental unresponsiveness and is a potential predictor of individual differences in development.


The Australian Mohs database: periocular squamous cell carcinoma.
To present the results of all patients with periocular squamous cell carcinoma (SCC) treated with Mohs' micrographic surgery (MMS) in Australia between 1993 and 1999. Prospective, noncomparative, multicenter, interventional case series. Seventy-nine patients undergoing MMS for periocular SCC over a 7-year period (1993-1999). Recurrence after MMS, site, size, prior recurrence, defect size, histologic subtype, and presence of perineural invasion (PNI). Seventy-nine patients had 54 (68%) lower eyelid, 19 (24%) medial canthus, and 6 (7.6%) upper eyelid SCCs. The most common histologic subtypes were well differentiated (48.7%) and moderately differentiated (35.1%) SCC. Seventy-three percent were primary tumors and 27% were recurrent tumors. Three (4.3%) cases had histologically confirmed PNI. Two of the 3 cases with PNI were recurrent (P = 0.1355), and surgical excision was the most frequent prior treatment (P = 0.0192), with up to 3 prior excisions in 1 case. None of the patients with PNI received adjunctive radiotherapy, and none have recurred with follow-ups to date of 4, 5, and 6 years, respectively. A median follow-up of 73 months (mean, 77.3; range, 42-117) was available in 56 (71%) cases, of which 2 (3.64%; exact 95% confidence interval, 0.44%-12.53%) recurred. Both cases were primary, moderately differentiated SCCs. This is the largest reported prospective series of periocular SCC managed by MMS, with a median follow-up of over 5 years. It demonstrates that MMS has the lowest reported recurrence rate (3.64%) of any treatment modality for periocular SCC. That there were no recurrences for cases with PNI further emphasizes the importance of margin-controlled excision for periocular SCC.


A scaffold containing zinc oxide for Schwann cell-mediated axon growth.
Objective.Schwann cells (SCs) transplanted in damaged nervous tissue promote axon growth, which may support the recovery of function lost after injury. However, SC transplant-mediated axon growth is often limited and lacks direction.Approach.We have developed a zinc oxide (ZnO) containing fibrous scaffold consisting of aligned fibers of polycaprolactone (PCL) with embedded ZnO nanoparticles as a biodegradable, bifunctional scaffold for promoting and guiding axon growth. This scaffold has bifunctional properties wherein zinc is released providing bioactivity and ZnO has well-known piezoelectric properties where piezoelectric materials generate electrical activity in response to minute deformations. In this study, SC growth, SC-mediated axon extension, and the presence of myelin basic protein (MBP), as an indicator of myelination, were evaluated on the scaffolds containing varying concentrations of ZnOin vitro. SCs and dorsal root ganglion (DRG) neurons were cultured, either alone or in co-culture, on the scaffolds.Main results.Findings demonstrated that scaffolds with 1 wt.% ZnO promoted the greatest SC growth and SC-mediated axon extension. The presence of brain-derived neurotrophic factor (BDNF) was also determined. BDNF increased in co-cultures for all scaffolds as compared to SCs or DRGs cultured alone on all scaffolds. For co-cultures, cells on scaffolds with low levels of ZnO (0.5 wt.% ZnO) had the highest amount of BDNF as compared to cells on higher ZnO-containing scaffolds (1 and 2 wt.%). MBP immunostaining was only detected in co-cultures on PCL control scaffolds (without ZnO).Significance.The results of this study demonstrate the potential of the ZnO-containing scaffolds for SC-mediated axon growth and its potential for use in nervous tissue repair.


Pressure ulcer prevention in nursing homes: nurse descriptions of individual and organization level factors.
Sustaining pressure ulcer prevention (PUP) in nursing homes has been difficult to achieve. Implementation science researchers suggest that identification of individual staff and organizational factors influencing current practices is essential to the development of an effective and customized plan to implement practice changes in a specific setting. A mixed methods approach was used to describe nurses' perceptions of individual and organization-level factors influencing performance of PUP in two Veterans Health Administration (VHA) nursing homes prior to implementation of a national VHA initiative on Hospital Acquired Pressure Ulcers (HAPUs). Individual interviews of 16 nursing staff were conducted. Individual factors influencing practice were a personal sense of responsibility to Veterans and belief in the effectiveness and importance of preventive measures. Organizational factors were existence of cooperative practices between nursing assistants and licensed nurses in assessing risk; teamwork, communication, and a commitment to Veterans' well-being. Integration and reinforcement of such factors in the development and maintenance of customized plans of PUP initiatives is recommended.


The Nurse Practitioner Workforce in Western Canada: A Cross-Sectional Practice Analysis Comparison.
Regular examination of health workforce data is essential given the pace of health system and legislative changes. Health workforce studies pertaining to nurse practitioner (NP) practice are needed to examine the gaps between work activities, policy, human resource supply, or for population needs. Jurisdictional comparison studies can provide essential information about NP practice for governments to respond to health workforce deficiencies or engage in service planning. In Canada, there is limited provincial-territorial jurisdictional NP workforce data to support health planning or policy change. This descriptive cross-sectional study was to examine the similarities and differences in practice patterns of Canadian NPs. In 2016 and 2017, an electronic survey was sent to all 852 registered NPs in three Canadian provinces, yielding a large convenience sample of 375 NP respondents. The results of this study underscore the value of NPs' extensive registered nurse expertize as well as their ability to serve diverse patient populations, work in varied healthcare settings, and provide care to medically complex patients. The study findings also show that NPs in all three jurisdictions work to their full scope of practice, in both rural and urban settings. This study is the first to compare NP workforce data across multiple Canadian jurisdictions simultaneously. Studies of this type are valuable tools for understanding the demographics, education, integration, and employment activities of NPs and can aid governments in addressing workforce planning.


Does Kinesiology Taping Improve Muscle Strength and Function in Knee Osteoarthritis? A Single-Blind, Randomized and Controlled Study.
This study aims to evaluate the efficacy of kinesiology taping (KT) on pain, movement restriction, walking speed, daily living activities, and isokinetic muscle performance in female patients with knee osteoarthritis. The study included 61 female patients (mean age 53.5±3.5 years; range 50 to 60 years) who were admitted to Physical Medicine and Rehabilitation Outpatient Clinic with complaints of mechanical knee pain. Knee osteoarthritis was diagnosed according to the clinical/ radiological criteria of the American College of Rheumatology and patients with Kellgren-Lawrence grade 2 and 3 were included. Patients were randomly divided into two groups. Both groups received transcutaneous electrical nerve stimulation for 30 minutes, hot pack for 30 minutes, and therapeutic ultrasound for 10 minutes in a day for three weeks (five days in a week). Also, one group was treated with KT (KT group, n=31) while the other group was treated with sham-KT (sham-KT group, n=30) one time a week for three weeks. A home program of around the knee strengthening exercises was recommended for all patients. All patients were evaluated with visual analog scale, Western Ontario and McMaster Osteoarthritis Index (WOMAC) values, goniometric measurement of active knee range of motion, 50-meter walking distance, and isokinetic knee extensor muscle peak torque measurements before treatment, at the end of treatment, and at one month and three months after treatment. There was a significant improvement in after treatment and first month visual analog scale values in the KT group compared to the sham-KT group (p<0.05). In the KT group, the WOMAC pain and WOMAC total scores decreased significantly after treatment compared to the sham-KT group (p<0.05). Visual analog scale values were significantly decreased after treatment in both groups (p<0.05). In both groups; WOMAC pain, stiffness, physical function, and total values decreased significantly after treatment (p<0.05). In both groups, isokinetic quadriceps peak torque measurements were increased after treatment (p<0.05). The application of KT to females with knee osteoarthritis appears to be a method that may be effective on pain and functional capacity.


Current progress and future perspectives in the development of anti-polo-like kinase 1 therapeutic agents.
Although significant levels of side effects are often associated with their use, microtubule-directed agents that primarily target fast-growing mitotic cells have been considered to be some of the most effective anti-cancer therapeutics. With the hope of developing new-generation anti-mitotic agents with reduced side effects and enhanced tumor specificity, researchers have targeted various proteins whose functions are critically required for mitotic progression. As one of the highly attractive mitotic targets, polo-like kinase 1 (Plk1) has been the subject of an extensive effort for anti-cancer drug discovery. To date, a variety of anti-Plk1 agents have been developed, and several of them are presently in clinical trials. Here, we will discuss the current status of generating anti-Plk1 agents as well as future strategies for designing and developing more efficacious anti-Plk1 therapeutics.


Tracking Low-Copy Transcription Factors in Living Bacteria: The Case of the lac Repressor.
Transcription factors control the expression of genes by binding to specific sites in DNA and repressing or activating transcription in response to stimuli. The lac repressor (LacI) is a well characterized transcription factor that regulates the ability of bacterial cells to uptake and metabolize lactose. Here, we study the intracellular mobility and spatial distribution of LacI in live bacteria using photoactivated localization microscopy combined with single-particle tracking. Since we track single LacI molecules in live cells by stochastically photoactivating and observing fluorescent proteins individually, there are no limitations on the copy number of the protein under study; as a result, we were able to study the behavior of LacI in bacterial strains containing the natural copy numbers (∼40 monomers), as well as in strains with much higher copy numbers due to LacI overexpression. Our results allowed us to determine the relative abundance of specific, near-specific, and non-specific DNA binding modes of LacI in vivo, showing that all these modes are operational inside living cells. Further, we examined the spatial distribution of LacI in live cells, confirming its specific binding to lac operator regions on the chromosome; we also showed that mobile LacI molecules explore the bacterial nucleoid in a way similar to exploration by other DNA-binding proteins. Our work also provides an example of applying tracking photoactivated localization microscopy to studies of low-copy-number proteins in living bacteria.


Triple immunocytochemical staining in fine-needle aspirate of a neck mass: a case report and review of the literature.
Immunocytochemistry involving single or double antibodies can be used to support or confirm a diagnosis in difficult cytology cases. There are a few reports on the application of a cocktail of 3 antibodies in the literature. We would like to report our experience using this technique in a difficult case of cervical lymph node fine-needle aspiration cytology (FNAC). A 75-year-old woman presented with a left neck mass for which FNAC was performed. The aspirates were cellular comprising atypical cells present in loose aggregates. The cells exhibited an ovoid and spindled morphology with no evidence of keratinization or any other differentiating features. The cytology was suggestive of metastatic poorly differentiated neoplasm; however, the presence of ovoid histiocytic-looking cells required further assessment. Triple immunostaining using a cocktail of CK5/6, CK34, and CD68 was performed. The spindled atypical cells were positive for CK5/6 and CK34 (brown cytoplasmic staining) and the ovoid histiocytic-looking cells were positive for CD68 (red cytoplasmic staining). Based on immunostaining, the cytological diagnosis of metastatic squamous cell carcinoma was confirmed. In view of our findings, the utility of triple immunocytochemical staining in cytology smears of poorly differentiated malignancy and a brief review of the literature are highlighted in this report.


Genome-wide Association Analysis of Eye Movement Dysfunction in Schizophrenia.
Eye movements are considered endophenotypes of schizophrenia. However, the genetic factors underlying eye movement are largely unknown. In this study, we explored the susceptibility loci for four eye movement scores: the scanpath length during the free viewing test (SPL), the horizontal position gain during the fast Lissajous paradigm of the smooth pursuit test (HPG), the duration of fixations during the far distractor paradigm of the fixation stability test (DF) and the integrated eye movement score of those three scores (EMS). We found 16 SNPs relevant to the HPG that were located in 3 genomic regions (1q21.3, 7p12.1 and 20q13.12) in the patient group; however, these SNPs were intronic or intergenic SNPs. To determine whether these SNPs occur in functional non-coding regions (i.e., enhancer or promoter regions), we examined the chromatin status on the basis of publicly available epigenomic data from 127 tissues or cell lines. This analysis suggested that the SNPs on 1q21.3 and 20q13.12 are in enhancer or promoter regions. Moreover, we performed an analysis of expression quantitative trait loci (eQTL) in human brain tissues using a public database. Finally, we identified significant eQTL effects for all of the SNPs at 1q21.3 and 20q13.12 in particular brain regions.


Li, P HY-021068 alleviates cerebral ischemia-reperfusion injury by inhibiting NLRP1 inflammasome and restoring autophagy function in mice.
Cerebral ischemia-reperfusion injury (CIRI) is a severe pathological condition that involves oxidative stress, inflammatory response, and neuronal damage. HY-021068 belongs to a new drug of chemical class 1, which is a potential thromboxane synthase inhibitor. Our preliminary experiment found that HY-021068 has significant anti-neuroinflammatory and neuroprotective effects. However, the protective effect and mechanism of HY-021068 in CIRI remain unclear. To investigate the protective effect and mechanism of HY-021068 in CIRI mice. In mice, CIRI was induced by bilateral common carotid artery occlusion and reperfusion. Mice were treated with HY-021068 or LV-NLRP1-shRNA (lentivirus-mediated shRNA transfection to knock down NLRP1 expression). The locomotor activity, neuronal damage, pathological changes, postsynaptic density protein-95 (PSD-95) expression, NLRP1 inflammasome activation, autophagy markers, and apoptotic proteins were assessed in CIRI mice. In this study, treatment with HY-021065 and LV-NLRP1-shRNA significantly improved motor dysfunction and neuronal damage after CIRI in mice. HY-021065 and NLRP1 knockdown significantly ameliorated the pathological damage and increased PSD-95 expression in the cortex and hippocampus CA1 and CA3 regions. The further studies showed that compared with the CIRI model group, HY-021065 and NLRP1 knockdown treatment inhibited the expressions of NLRP1, ASC, caspase-1, and IL-1β, restored the expressions of p-AMPK/AMPK, Beclin1, LC3II/LC3I, p-mTOR/m-TOR and P62, and regulated the expressions of BCL-2, Caspase3, and BAX in brain tissues of CIRI mice in CIRI mice. These results suggest that HY-021068 exerts a protective role in CIRI mice by inhibiting NLRP1 inflammasome activation and regulating autophagy function and neuronal apoptosis. HY-021068 is expected to become a new therapeutic drug for CIRI.


The mechanism of μ-opioid receptor (MOR)-TRPV1 crosstalk in TRPV1 activation involves morphine anti-nociception, tolerance and dependence.
Initiated by the activation of various nociceptors, pain is a reaction to specific stimulus modalities. The μ-opioid receptor (MOR) agonists, including morphine, remain the most potent analgesics to treat patients with moderate to severe pain. However, the utility of MOR agonists is limited by the adverse effects associated with the use of these drugs, including analgesic tolerance and physical dependence. A strong connection has been suggested between the expression of the transient receptor potential vanilloid type 1 (TRPV1) ion channel and the development of inflammatory hyperalgesia. TRPV1 is important for thermal nociception induction, and is mainly expressed on sensory neurons. Recent reports suggest that opioid or TRPV1 receptor agonist exposure has contrasting consequences for anti-nociception, tolerance and dependence. Chronic morphine exposure modulates TRPV1 activation and induces the anti-nociception effects of morphine. The regulation of many downstream targets of TRPV1 plays a critical role in this process, including calcitonin gene-related peptide (CGRP) and substance P (SP). Additional factors also include capsaicin treatment blocking the anti-nociception effects of morphine in rats, as well as opioid modulation of TRPV1 responses through the cAMP-dependent PKA pathway and MAPK signaling pathways. Here, we review new insights concerning the mechanism underlying MOR-TRPV1 crosstalk and signaling pathways and discuss the potential mechanisms of morphine-induced anti-nociception, tolerance and dependence associated with the TRPV1 signaling pathway and highlight how understanding these mechanisms might help find therapeutic targets for the treatment of morphine induced antinociception, tolerance and dependence.


[Waist-hip ratio and perioperative bleeding in patients who underwent radical prostatectomy].
Radical prostatectomy is associated with perioperative bleeding and multiple transfusions. Abdominal obesity is a perioperative risk factor. We suggest that the adipocytes have a protective effect in oncological patients undergoing radical prostatectomy. The aim was to evaluate the effect of waist-hip ratio (WHR) on the amount of bleeding and perioperative transfusion requirements in oncological patients undergoing radical prostatectomy. We performed a cohort study in 156 patients. We had two groups: the control group (WHR<0.95) and the problem group (WHR≥0.95). Blood loss and fractions transfused during surgery and in the postoperative period were recorded. In the analysis of variables, for descriptive statistics we used measures of central tendency and dispersion. Inferential statistics was obtained by chi square, Student's t test, Mann-Whitney U and ANOVA. A p<0.05 was significant. We found significant differences in weight, body mass index, waist, WHR, perioperative bleeding, fractions transfused, permanence of the catheter, and hospital days. Patients who underwent radical prostatectomy with a WHR≥0.95 had a magnitude of perioperative bleeding and transfusion requirements with a WHR<0.95.


Thoracoscopic lobectomy: comparison of intraoperative and postoperative outcomes between 3 and 4 incision accesses.
Several techniques have been proposed to perform a video-assisted thoracic lobectomy. We compared the results of a 3 versus 4-port procedure, analyzing intraoperative data, morbidity, and mortality. Prospective analysis of 30 consecutive patients who underwent a 4-port approach video-assisted thoracic lobectomy (group A) and comparison with a historical series with 30 patients who had a 3-port video-assisted thoracic lobectomy (group B). The groups were comparable for clinical characteristics and pathological staging. There was no difference in operating time: median, 128 min for group A versus 129 min for group B (P = 0.9). There was a significant difference in rate of conversion to thoracotomy: 1 of 30 (3.3%) in group A and 7 of 30 (23.3%) in group B (3 ports) (P = 0.02). In group A, 11 patients (36.7%) experienced postoperative complications and in group B, 13 patients (43.3%; P = 0.6). The difference in median time to drain removal and median length of hospital stay between the two groups was not significant. There was a significant difference in persistent pain between group A and group B: 6 patients (20%) in group B presented with persistent neuropathic pain on regular medication (P = 0.02). Our study showed that the 4-port approach was similar in operative time, length of drain and hospital stay but showed a statistically significant lower conversion rate and lower rate of persistent pain than the 3-port access.


Femoral fracture type can be predicted from femoral structure: A finite element study validated by digital volume correlation experiments.
Proximal femoral fractures can be categorized into two main types: Neck and intertrochanteric fractures accounting for 53% and 43% of all proximal femoral fractures, respectively. The possibility to predict the type of fracture a specific patient is predisposed to would allow drug and exercise therapies, hip protector design, and prophylactic surgery to be better targeted for this patient rendering fracture preventing strategies more effective. This study hypothesized that the type of fracture is closely related to the patient-specific femoral structure and predictable by finite element (FE) methods. Fourteen femora were DXA scanned, CT scanned, and mechanically tested to fracture. FE-predicted fracture patterns were compared to experimentally observed fracture patterns. Measurements of strain patterns to explain neck and intertrochanteric fracture patterns were performed using a digital volume correlation (DVC) technique and compared to FE-predicted strains and experimentally observed fracture patterns. Although loaded identically, the femora exhibited different fracture types (six neck and eight intertrochanteric fractures). CT-based FE models matched the experimental observations well (86%) demonstrating that the fracture type can be predicted. DVC-measured and FE-predicted strains showed obvious consistency. Neither DXA-based BMD nor any morphologic characteristics such as neck diameter, femoral neck length, or neck shaft angle were associated with fracture type. In conclusion, patient-specific femoral structure correlates with fracture type and FE analyses were able to predict these fracture types. Also, the demonstration of FE and DVC as metrics of the strains in bones may be of substantial clinical value, informing treatment strategies and device selection and design. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:993-1001, 2018.


Cyclin D1 regulates osteoarthritis chondrocyte apoptosis via WNT3/β-catenin signalling.
Objective: Cyclin D1 was an important molecular involved in the pathological process of osteoarthritis (OA). The purpose of this study was to identify the effect and potential mechanism of Cyclin D1 for the proliferation and apoptosis of OA chondrocytes. Methods: We used polymerase chain reaction (PCR) method to identify the expression levels of Cyclin D1 and down-stream Wnt/β-catenin pathway-related genes in OA chondrocytes according to the grade of OA. Small interfering RNA (siRNA) or overexpression of Cyclin D1 were used to identify the role of Cyclin D1 in cell proliferation and apoptosis. Next, we used XAV-939 to inhibit the Wnt/β-catenin pathway and explore the relevant mechanism. Results: Cyclin D1 was significantly decreased with OA grade (p < .05). After siCyclin D1 transfection, the expression level of WNT3 and nuclear β-catenin were significantly increased, while Wnt10a and total β-catenin were not obviously changed. Co-cultured with XAV-939 and siCyclin D1 abolished the effects of siCyclin D1 on proliferation and apoptosis of OA chondrocytes (p < .05). Conclusions: Cyclin D1 regulated chondrocyte proliferation and apoptosis through Wnt3/β-catenin instead of Wnt10a/β-catenin signalling pathway.


Can Stochastic Pacing Restore Heart Rate Variability in Diseased Hearts? An In-vivo Ovine Case Study.
Heart rate variability (HRV) is an important clinical parameter that depicts the autonomic balance. Diminished HRV has been associated with diseased hearts and incorporating stochasticity in pacing has been investigated as a potential mechanism for restoring the altered autonomic balance and preventing cardiac arrhythmias. We studied the change in HRV with the development of chronic myocardial infarction (MI) in adult sheep (n=16). Next, we investigated the utility of stochastic pacing in modulating HRV in-vivo in both sham and MI hearts. The propensity of the heart to the development of cardiac alternans, a known precursor to tachyarrhythmias, was studied under three different pacing techniques, namely periodic pacing, stochastic pacing and constant diastolic interval (DI) pacing in one sham and one MI sheep. Autonomic balance was observed to be altered after 6 weeks of chronic MI. Increased heart rate, QTc interval, standard deviation of the R-R intervals and LF/HF ratio was observed in MI hearts. Stochastic pacing was found to be proarrhythmic and increased T-wave alternans burden was observed with increase in stochasticity. Maintaining a constant DI on every beat demonstrated reduced alternans levels compared to both periodic and stochastic pacing.Clinical Relevance-Our results demonstrate that precise control of the diastolic interval may be more beneficial in inhibiting arrhythmias than stochastic pacing.


Surface properties and hemocompatibility of alkyl-siloxane monolayers supported on silicone rubber: effect of alkyl chain length and ionic functionality.
Self-assembled monolayers of alkylsiloxanes supported on poly(dimethylsiloxane) (PDMS) rubber were used as model systems to study the relation between blood compatibility and surface composition. The inner lumen of PDMS tubes were first treated with an oxygen plasma. The resultant oxidized surfaces were post-derivatized by reaction with alkyltrichlorosilanes to form the monolayer films. The alkyl chain lengths used were slightly longer than in a previous study, and this may alter the phase-state of the monolayer from liquid-like to crystalline. The chemical properties of the monolayer were controlled by varying the chemical composition of the alkyltrichlorosilanes used. Terminal functionalities included -CH3, -CF3, -COOH, -SO3H and -(CH2CH2O)4OH. Surface derivatization was verified with static contact angle measurements and X-ray photoelectron spectroscopy. Blood compatibility was evaluated using a canine ex vivo arterio-venous series shunt model. Surfaces grafted with hydrophobic head groups such as -CH3 and -CF3 were significantly less thrombogenic than the surfaces composed of ionic head groups such as -COOH and -SO3H. Surfaces enriched in -(CH2CH2O)4OH had an intermediate thrombogenicity. Silastic pump grade tubing and polyethylene tubing, used as controls, were found to be the least thrombogenic of all the surfaces tested.


Simultaneous sequencing of 24 genes associated with steroid-resistant nephrotic syndrome.
Up to 95% of children presenting with steroid-resistant nephrotic syndrome in early life will have a pathogenic single-gene mutation in 1 of 24 genes currently associated with this disease. Others may be affected by polymorphic variants. There is currently no accepted diagnostic algorithm for clinical genetic testing. The hypothesis was that the increasing reliability of next generation sequencing allows comprehensive one-step genetic investigation of this group and similar patient groups. This study used next generation sequencing to screen 446 genes, including the 24 genes known to be associated with hereditary steroid-resistant nephrotic syndrome. The first 36 pediatric patients collected through a national United Kingdom Renal Registry were chosen with comprehensive phenotypic detail. Significant variants detected by next generation sequencing were confirmed by conventional Sanger sequencing. Analysis revealed known and novel disease-associated variations in expected genes such as NPHS1, NPHS2, and PLCe1 in 19% of patients. Phenotypically unexpected mutations were also detected in COQ2 and COL4A4 in two patients with isolated nephropathy and associated sensorineural deafness, respectively. The presence of an additional heterozygous polymorphism in WT1 in a patient with NPHS1 mutation was associated with earlier-onset disease, supporting modification of phenotype through genetic epistasis. This study shows that next generation sequencing analysis of pediatric steroid-resistant nephrotic syndrome patients is accurate and revealing. This analysis should be considered part of the routine genetic workup of diseases such as childhood steroid-resistant nephrotic syndrome, where the chance of genetic mutation is high but requires sequencing of multiple genes.


Random mutagenesis to identify novel Helicobacter mustelae virulence factors.
Helicobacter mustelae is a gastric pathogen of ferrets, where it causes disorders similar to those caused by Helicobacter pylori in humans. The H. mustelae ferret model therefore has potential for the in vivo study of Helicobacter pathogenesis in general. In this study a library of 500 individual H. mustelae mutants was generated using an in vitro random insertion mutagenesis technique. Mutants were subsequently tested for motility and adherence, and 43 of the 500 mutants tested were found to be nonmotile in a soft agar assay. Of these 43 mutants, seven were subsequently identified as deficient in their ability to adhere to AGS cells. Insertion had taken place in different positions in the H. mustelae genome, and included mutants in or near to genes involved in motility and urease activity (e.g. the chemotaxis gene cheV and the urease accessory gene ureH). The development of a mutant library for a natural animal model of Helicobacter infection provides the opportunity to study in vivo the role of candidate Helicobacter virulence genes.


Tools for probing local circuits: high-density silicon probes combined with optogenetics.
To understand how function arises from the interactions between neurons, it is necessary to use methods that allow the monitoring of brain activity at the single-neuron, single-spike level and the targeted manipulation of the diverse neuron types selectively in a closed-loop manner. Large-scale recordings of neuronal spiking combined with optogenetic perturbation of identified individual neurons has emerged as a suitable method for such tasks in behaving animals. To fully exploit the potential power of these methods, multiple steps of technical innovation are needed. We highlight the current state of the art in electrophysiological recording methods, combined with optogenetics, and discuss directions for progress. In addition, we point to areas where rapid development is in progress and discuss topics where near-term improvements are possible and needed.


Ethical issues and addiction.
The epidemic of substance abuse continues to pose a significant challenge to clinicians nationwide. Although there is a tendency to simply associate drug abuse with poverty, the problem affects every social stratum gender and race; and pregnant women are no exception. Caring for pregnant, substance-using women and their infants presents complex legal and ethical issues. Debate is ongoing about whether criminal penalties should be imposed on women based solely on their use of alcohol and other drugs during pregnancy. Furthermore, controversies persist about the rights and wishes of pregnant women versus the interests of their fetuses. For health professionals, conflict arises when the pregnant woman chooses behaviors that have the potential to harm the developing fetus. The ethical dilemma arises from competing autonomy-based and beneficence-based obligations to the maternal-fetal dyad. This chapter explores the ethics-based conflicts in the delivery of health care to drug abusing pregnant women.


Influence of Calcium Perchlorate on the Search for Martian Organic Compounds with MTBSTFA/DMF Derivatization.
N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA), mixed with the solvent N,N-dimethylformamide (DMF), is used as a derivatizing reagent by the Sample Analysis at Mars (SAM) experiment onboard NASA's Curiosity rover and will soon be utilized by the Mars Organic Molecule Analyzer experiment onboard the ESA/Roscosmos Rosalind Franklin rover. The pyrolysis products of MTBSTFA, DMF, and the MTBSTFA/DMF mixtures, obtained at different temperatures, were analyzed. Two different pyrolysis modes were studied, flash pyrolysis and ramp pyrolysis (35°C/min), to evaluate the potential influence of the sample heating speed on the production of products in space chromatographs. The effect of the presence of calcium perchlorate on the pyrolysis products of MTBSTFA/DMF was also studied to ascertain the potential effect of perchlorate species known to be present at the martian surface. The results show that MTBSTFA/DMF derivatization should be applied below 300°C when using flash pyrolysis, as numerous products of MTBSTFA/DMF were formed at high pyrolysis temperatures. However, when an SAM-like ramp pyrolysis was applied, the final pyrolysis temperature did not appear to influence the degradation products of MTBSTFA/DMF. All products of MTBSTFA/DMF pyrolysis are listed in this article, providing a major database of products for the analysis of martian analog samples, meteorites, and the in situ analysis of martian rocks and soils. In addition, the presence of calcium perchlorate does not show any obvious effects on the pyrolysis of MTBSTFA/DMF: Only chloromethane and TBDMS-Cl (chloro-tertbutyldimethylsilane) were detected, whereas chlorobenzene and other chlorine-bearing compounds were not detected. However, other chlorine-bearing compounds were detected after pyrolysis of the Murchison meteorite in the presence of calcium perchlorate. This result reinforces previous suggestions that chloride-bearing compounds could be reaction products of martian samples and perchlorate.


Comparative effects of glyceryl trinitrate and amyl nitrite on pulse wave reflection and augmentation index.
The influence of vasodilators on augmentation index (AIx) offers a simple, rapid and noninvasive method of evaluating vascular function. Glyceryl trinitrate (GTN) is widely used as an endothelium-independent vasodilator, although other nitrates that are shorter acting may have advantages in clinical studies. The aim of this study was to compare the effects of two short-acting nitrates, GTN and amyl nitrite, which have differing pharmacodynamic profiles. Twenty-one healthy volunteers (15 male; mean age 35 years, range 21-56 years) attended on three occasions and received sublingual GTN (0.5 mg for 3 min), inhaled amyl nitrite (0.2 ml inhaled for 30 s), or no treatment in a randomized cross-over design. Haemodynamic responses of AIx, blood pressure and thoracic bioimpedance (heart rate, cardiac index) were assessed by measurement at baseline, every 60 s for the first 5 min, and then every 5 min for a further 55 min. AIx was reduced by amyl nitrite (peak effect -9 +/- 2% at 1 min, P < 0.002) and GTN (peak effect -12 +/- 3% at 4 min, P < 0.05). Compared with amyl nitrite, the onset and offset of action of GTN was slower. Amyl nitrite initially increased heart rate by 27 +/- 4% (P < 0.001) and cardiac index by 13 +/- 3% (P < 0.001) whereas GTN had no significant effect (P > 0.05). Neither agent affected blood pressure. GTN causes a slower and more sustained reduction in AIx than amyl nitrite. Although amyl nitrite causes a more rapid fall and recovery in AIx, it induces a reflex tachycardia that may limit interpretation of the initial (1 min) but not later (2 min) changes in AIx. The prolonged offset of GTN suggests that a sufficient washout period must be included when making repeated measures or when assessing the subsequent effects of other agents.


Key issues for estimating the impact and cost-effectiveness of seasonal influenza vaccination strategies.
Many countries have considered or are considering modifying their seasonal influenza immunization policies. Estimating the impact of such changes requires understanding the existing clinical and economic burden of influenza, as well as the potential impact of different vaccination options. Previous studies suggest that vaccinating clinical risk groups, health care workers, children and the elderly may be cost-effective. However, challenges in such estimation include: (1) potential cases are not usually virologically tested; (2) cases have non-specific symptoms and are rarely reported to surveillance systems; (3) endpoints for influenza proxies (such as influenza-like illness) need to be matched to case definitions for treatment costs, (4) disease burden estimates vary from year to year with strain transmissibility, virulence and prior immunity, (5) methods to estimate productivity losses due to influenza vary, (6) vaccine efficacy estimates from trials differ due to variation in subtype prevalence, vaccine match and case ascertainment, and (7) indirect (herd) protection from vaccination depends on setting-specific variables that are difficult to directly measure. Given the importance of knowing the impact of changes to influenza policy, such complexities need careful treatment using tools such as population-based trial designs, meta-analyses, time-series analyses and transmission dynamic models.


The Impact of Technology on People with Autism Spectrum Disorder: A Systematic Literature Review.
People with autism spectrum disorder (ASD) tend to enjoy themselves and be engaged when interacting with computers, as these interactions occur in a safe and trustworthy environment. In this paper, we present a systematic literature review on the state of the research on the use of technology to teach people with ASD. We reviewed 94 studies that show how the use of technology in educational contexts helps people with ASD develop several skills, how these approaches consider aspects of user experience, usability and accessibility, and how game elements are used to enrich learning environments. This systematic literature review shows that the development and evaluation of systems and applications for users with ASD is very promising. The use of technological advancements such as virtual agents, artificial intelligence, virtual reality, and augmented reality undoubtedly provides a comfortable environment that promotes constant learning for people with ASD.


Health Insurance and Long-Term Care Services for the Disabled Elderly in China: Based on CHARLS Data.
This paper aimed to explore the relationship between the different factors, especially health insurance, and the availability of long-term care (LTC) services, among the disabled elderly. Based on the data of China Health and Retirement Longitudinal Study (CHARLS), the logistic regression model was utilized to evaluate the influence of the different factors, especially health insurance, on the availability of long-term care services. Our findings show some interesting results. Firstly, the findings suggest that informal long-term care (LTC) services for elderly persons with disabilities heavily depend on a family member from different health insurance groups. About 80.733% of the disabled elderly depend on a family member as their primary caregivers. Secondly, other influence factors such as income and area of residence were also significantly related to the availability of long-term rental services. Thirdly, Health insurance is a very important factor influencing the availability of Long-term care services both in urban and rural areas (p<0.001) but Income is the most interesting variable. Based on our results, the growth and integration of formal long-term care (LTC) services should be facilitated. Firstly, policymakers can encourage formal long-term care (LTC) services from a variety of sources to work together to increase overall supply capability. Secondly, the long-term living security needs of people who do not have health insurance should be regulated through subsidies according to the economic status.


The treatment of complex regional pain syndrome type I with free radical scavengers: a randomized controlled study.
To compare the effects of two free radical scavengers, dimethylsulfoxide 50% (DMSO) and N-acetylcysteine (NAC), for treatment of complex regional pain syndrome I (CRPS I), a randomized, double-dummy controlled, double-blind trial was conducted. Two outpatient clinics of two university hospitals in The Netherlands participated in the study and 146 patients, were included over a period of 24 months. Patients were randomized into two treatment groups, one was instructed to apply DMSO 50% five times daily to the affected extremity, the second was treated with NAC 600mg effervescent tablets three times daily, both combined with placebo. Interventions were accompanied by pain medication, occupational therapy for upper extremity CRPS I and physical therapy for lower extremity CRPS I in specific circumstances. Treatment was given for 17 weeks, with a possibility to continue or switch medication after this period, up to 1 year following the onset of treatment. An impairment level sum score was the primary outcome measure. Upper and lower extremity skills and functions, and general health status were also evaluated. Overall, no significant differences were found between NAC and DMSO after 17 and 52 weeks on impairment level and general health status. Significant differences were found for subscores of lower extremity function, in favor of DMSO-treatment. Subgroup analysis showed more favorable results for DMSO for warm CRPS I and significantly better performance of NAC for patients with a cold CRPS I. Results tended to be negatively influenced if the duration of the complaint was longer. Treatment with DMSO and NAC are generally equally effective in treatment of CRPS I. Strong indications exist for differences in effects for subgroups of patients with warm or cold CRPS I: for warm CRPS I, DMSO-treatment appears more favorable, while for cold CRPS I, NAC-treatment appears to be more effective.


Luminescent Mechanochromic 9-Anthryl Gold(I) Isocyanide Complex with an Emission Maximum at 900 nm after Mechanical Stimulation.
Upon mechanical stimulation, 9-anthryl gold(I) isocyanide complex 3 exhibited a bathochromic shift of its emission color from the visible to the infrared (IR) region, which is unprecedented in its magnitude. Prior to exposure to the mechanical stimulus, the polymorphs 3α and 3β exhibit emission wavelength maxima (λem,max) at 448 and 710 nm, respectively. Upon grinding, the λem,max of 3αground and 3βground are bathochromically shifted to 900 nm, i.e., Δλem,max (3α) = 452 nm or 1.39 eV. Polymorphs 3α and 3β thus represent the first examples of mechanochromic luminescent materials with λem,max in the IR region.


CD46 Contributes to the severity of group A streptococcal infection.
Streptococcus pyogenes (group A Streptococcus) is a human pathogen that causes a wide variety of diseases ranging from uncomplicated superficial infections to severe infections such as streptococcal toxic shock syndrome and necrotizing fasciitis. These bacteria interact with several host cell receptors, one of which is the cell surface complement regulator CD46. In this study, we demonstrate that infection of epithelial cells with S. pyogenes leads to the shedding of CD46 at the same time as the bacteria induce apoptosis and cell death. Soluble CD46 attached to the streptococcal surface, suggesting that bacteria might bind available extracellular CD46 as a strategy to survive and avoid host defenses. The protective role of human CD46 was demonstrated in ex vivo whole-blood assays showing that the growth of S. pyogenes was enhanced in blood from mice expressing human CD46. Finally, in vivo experimental infection showed that bacteremia levels, arthritis frequency, and mortality were higher in CD46 transgenic mice than in nontransgenic mice. Taken together, these results argue that bacterial exploitation of human CD46 enhances bacterial survival and represents a novel pathogenic mechanism that contributes to the severity of group A streptococcal disease.


[New approaches in the treatment of speech disorders in children with an organic brain lesion].
Speech disorders encountered in children presenting with neurological pathology make their medical and social rehabilitation a real challenge. Overall sixty children were examined. Of these, 38 had infantile cerebral paralysis, 22 that being presented with sequelae of the hypoxic affection of the brain in ante- and perinatal period, who exhibited speech disorders (alalia, dysarthria, retarded speech development). Described in the paper are features of bloodflow disorders in intracranial and vertebral arteries as evidenced by ultrasonic Doppler technique and changes in brain bioelectrical activity according to findings from electroencephalomapping. In 53 patients, a positive effect was obtained as a result of therapy conducted in accordance with the stimulation type techniques (electroscalp therapy against the background of intramuscular administration of nicotinic acid plus laser therapy, transcutaneous electrostimulation, employment of such drug preparations as cogitum, nero-force, sirdalud).


Training spatial skills in men and women.
Recent studies suggest that even short-term video game training may transfer to other cognitive tasks. With the popularity of the Nintendo Wii with women, more of them might be exposed to the games that will increase their mental rotation skills. Because performance on mental rotation tests (MRT) has been linked to math performance in women, and thus may ultimately contribute to the under representation of women in STEM fields, it is important to continue to explore ways to decrease or eliminate the robust sex difference in mental rotation. The present study of 30 men and 30 women provides additional evidence that women may benefit from short-term (1 hour) training on either a Nintendo Wii™ or GameCube console to increase their mental rotation skills. One hour of video game training not only increased women's MRT scores to a level similar to men's scores, but also produced greater average improvement for women, even when controlling for experiential factors such as spatial and masculine childhood activities that could contribute to the sex difference in spatial ability.


Structural dynamics of actin during active interaction with myosin: different effects of weakly and strongly bound myosin heads.
We have used optical spectroscopy (transient phosphorescence anisotropy, TPA, and fluorescence resonance energy transfer, FRET) to detect the effects of weakly bound myosin S1 on actin during the actomyosin ATPase cycle. The changes in actin were reported by (a) a phosphorescent probe (ErIA) attached to Cys 374 and (b) a FRET donor-acceptor pair, IAEDANS attached to Cys 374 and a nucleotide analogue (TNPADP) in the nucleotide-binding cleft. Strong interactions were detected in the absence of ATP, and weak interactions were detected in the presence of ATP or its slowly hydrolyzed analogue ATP-gamma-S, under conditions where a significant fraction of weakly bound acto-S1 complex was present and the rate of nucleotide hydrolysis was low enough to enable steady-state measurements. The results show that actin in the weakly bound complex with S1 assumes a new structural state in which (a) the actin filament has microsecond rotational dynamics intermediate between that of free actin and the strongly bound complex and (b) S1-induced changes are not propagated along the actin filament, in contrast to the highly cooperative changes due to the strongly bound complex. We propose that the transition on the acto-myosin interface from weak to strong binding is accompanied by transitions in the structural dynamics of actin parallel to transitions in the dynamics of interacting myosin heads.


Estimating time evolving cross-dependence of porcine reproduction and respiratory syndrome sero-prevalence in Danish swine herds.
Porcine reproductive and respiratory syndrome (PRRS) has been a challenge for the Danish swine industry. It is important to identify forms of time-dependence in PRRS sero-prevalence among swine herds at country level, in order to allocate resources in an effective manner. This is of particular value if an increase of PRRS sero-prevalence occurs in a particular type of herds and control measures are taken to avoid disease spread downwards the swine production system. The objective of this study was to describe the presence (or lack thereof) of the linear cross-dependence of PRRS sero-prevalence in Danish swine herds from 2012 to 2014. These temporal dependencies were calculated for Danish swine herds with different biosecurity status and directional trade contacts between them: from nucleus/breeding herds to multipliers and finisher herds and from nucleus/breeding herds to finishers via multiplier herds. We used a method proposed for non-stationary time-series decomposition which allows an assessment of the existence of linear cross-dependencies between multiple of types of herds. The results demonstrate the existence of cyclic patterns of direct linear positive and negative dependence between PRRS sero-prevalence between multipliers and finishers herds. This suggests that the associations between PRRS sero-prevalence time-series follow the swine production system downwards for the majority of the study period, i.e. an increase or decrease in PRRS sero-prevalence in multiplier is followed by a shift in PRRS sero-prevalence in finisher herds after a few weeks. For specific time periods, however, these associations might have been influenced by other factors.


[Detection of pulmonary tuberculosis in patients with mental disorders].
X-ray study is the main way of detecting tuberculosis in patients with mental disorsers. As dysadaptation progresses in the mentally ill, the detection rate of lung tissue destruction increases with the simultaneous reduction in the number of those discharging bacteria and the role of medical attendants increases in the goal-oriented selection of patients for an in-depth examination for a tuberculous process.


[Assessment and implementation of guidelines].
The importance of guidelines increases continuously on the political level whereas on the user level reservations and uncertainty persist. Consequently guidelines are not considered as they should be. Guidelines will develop their effectiveness only if they are firmly implemented in the delivery process. Acceptance problems spring from problems in development and from aspects of dissemination and implementation not sufficiently considered so far. Therefore a lot of countries have developed quality criteria for guidelines and programmes for quality promotion. To further the use of guidelines in the care process aspects of dissemination and implementation have to be recognized even in the development process. This has not been recognized sufficiently so far. Implementation of guidelines is a systematic approach which has to be connected seamlessly with other activities (e. g. quality management). The implementation process should be accompanied and evaluated so that a continuous adjustment is possible. Most of the existing guideline programmes do not consider this sufficiently. The following contribution gives a survey of results and introduces means and instruments for assessment and implementation of guidelines.


Therapeutic potential of the plasma proteome.
Plasma contains numerous and diverse proteins with existing and potential therapeutic value. Plasma has been used clinically as both a source of purified derivatives for treating diseases such as hemophilia, and as a diagnostic medium. Recent research directed towards mining plasma's true potential takes advantage of state-of-the-art proteomic analytical methods to develop multi-protein, disease-specific biomarker panels to improve the reliability and specificity of diagnostics. Recombinant production and chromatographic purification methods are increasing the yield and safety of traditional plasma derivatives. Emerging cell-based technologies are being applied to discover novel protein activities and identify epitope-specific antibodies that may have clinical promise.


Characterization of Blood Outgrowth Endothelial Cells (BOEC) from Porcine Peripheral Blood.
The endothelium is a dynamic integrated structure that plays an important role in many physiological functions such as angiogenesis, hemostasis, inflammation, and homeostasis. The endothelium also plays an important role in pathophysiologies such as atherosclerosis, hypertension, and diabetes. Endothelial cells form the inner lining of blood and lymphatic vessels and display heterogeneity in structure and function. Various groups have evaluated the functionality of endothelial cells derived from human peripheral blood with a focus on endothelial progenitor cells derived from hematopoietic stem cells or mature blood outgrowth endothelial cells (or endothelial colony-forming cells). These cells provide an autologous resource for therapeutics and disease modeling. Xenogeneic cells may provide an alternative source of therapeutics due to their availability and homogeneity achieved by using genetically similar animals raised in similar conditions. Hence, a robust protocol for the isolation and expansion of highly proliferative blood outgrowth endothelial cells from porcine peripheral blood has been presented. These cells can be used for numerous applications such as cardiovascular tissue engineering, cell therapy, disease modeling, drug screening, studying endothelial cell biology, and in vitro co-cultures to investigate inflammatory and coagulation responses in xenotransplantation.


[The reinnervation of skeletal muscles].
Experiments were conducted to study different methods of reinnervation of predenervated muscle, for which purpose three groups of animals were used. The first group (reinnervation as to Brunelli) served as a kind of control. In the second group, side notches were cut in the nerve-neurotizator before inserting it into the muscle, in the third group, endomysium and perimysium were defibered at the site of insertion into the muscle of the nerve-neurotizator. The obtained results showed that in the second and third groups processes of reinnervation proceed faster by far and in a more efficient way. This is because the nerve-neurotizator has side cuttings in it, which fact promotes the ingrowth into the muscle tissue of a great many of nerve conductors, and because the skeletal muscle neurotropism gets enhanced at the expense of the connective tissue being formed in sites of defiberation of endomysium and perimysium. The secured results permit recommending the proposed ways and means for reinnervation of denervated muscles in a clinical setting as more efficacious than that by Brunelli.


Ceramide regulates interaction of Hsd17b4 with Pex5 and function of peroxisomes.
The sphingolipid ceramide regulates beta-oxidation of medium and long chain fatty acids in mitochondria. It is not known whether it also regulates oxidation of very long chain fatty acids (VLCFAs) in peroxisomes. Using affinity chromatography, co-immunoprecipitation, and proximity ligation assays we discovered that ceramide interacts with Hsd17b4, an enzyme critical for peroxisomal VLCFA oxidation and docosahexaenoic acid (DHA) generation. Immunocytochemistry showed that Hsd17b4 is distributed to ceramide-enriched mitochondria-associated membranes (CEMAMs). Molecular docking and in vitro mutagenesis experiments showed that ceramide binds to the sterol carrier protein 2-like domain in Hsd17b4 adjacent to peroxisome targeting signal 1 (PTS1), the C-terminal signal for interaction with peroxisomal biogenesis factor 5 (Pex5), a peroxin mediating transport of Hsd17b4 into peroxisomes. Inhibition of ceramide biosynthesis induced translocation of Hsd17b4 from CEMAMs to peroxisomes, interaction of Hsd17b4 with Pex5, and upregulation of DHA. This data indicates a novel role of ceramide as a molecular switch regulating interaction of Hsd17b4 with Pex5 and peroxisomal function.