Prognostic significance of iNOS in epithelial ovarian cancer.
To study the association of inducible nitric oxide synthase (iNOS) expression with clinicopathological factors and prognosis in epithelial ovarian cancer. The study included 301 patients with primary epithelial ovarian cancer. iNOS expression was evaluated by immunohistochemistry using a mouse monoclonal antibody. iNOS positivity was observed as granular deposits in the cancer cell cytoplasm. The mean percentage of iNOS-positive cells was 50% in primary tumors (n=301), and 62% in metastatic lesions (n=43). iNOS expression correlated significantly with histological subtype of the tumor, as high (> 70%) iNOS expression was observed in mucinous tumors (p=0.009). Poorly differentiated tumors showed a tendency to low (< or = 70%) iNOS expression but without statistical significance. Low iNOS expression associated also significantly with large primary residual tumor (p=0.007) and tumor recurrence (p=0.04). The 10-year prognosis of the patients with high iNOS expression was better in disease-related survival (DRS) (p=0.009). However, in multivariate analysis only FIGO stage, primary residual tumor, and grade of the tumor were independent prognostic factors for DRS, but not the iNOS expression. A major proportion of human epithelial ovarian cancers expressed iNOS. The positive expression was an indicator of better disease-related survival. However, iNOS positivity could not overcome the importance of clinicopathological factors in prediction of prognosis.


Effects of Pirfenidone and Collagen-Polyvinylpyrrolidone on Macroscopic and Microscopic Changes, TGF-β1 Expression, and Collagen Deposition in an Experimental Model of Tracheal Wound Healing.
Tracheal stenosis (TS) is a fibrosis originated by prolonged inflammation and increased transforming growth factor beta 1 (TGF-β1) expression and collagen deposition (CD) in the tracheal wound. Several wound-healing modulators (WHMs) have been used to modulate the tracheal healing process and prevent TS, but they have failed, justifying the need to evaluate alternative WHM. The pirfenidone (PFD) and collagen-polyvinylpyrrolidone (Collagen-PVP) decrease inflammation and fibrosis. This study assessed the effect of PFD administration and Collagen-PVP topical application on macroscopic and microscopic changes, TGF-β1 expression, and CD in an experimental model of tracheal wound healing. Forty Wistar rats underwent cervical tracheoplasty, were divided into 4 groups (n = 10), and were treated with different WHM: group I, saline solution (SS); group II, Collagen-PVP; group III, mitomycin C (MMC); and group IV, 40 mg/kg PFD. Four weeks after surgery, the macroscopic and microscopic changes, in situ TGF-β1 expression, and CD in posttracheoplasty scars were evaluated. The animals treated with Collagen-PVP and PFD developed less inflammation and fibrosis than animals in the other study groups (p < 0.05, Kruskal-Wallis) and, moreover, showed lower TGF-β1 expression and CD than animals in group I (p < 0.05, ANOVA and Tukey's test). In conclusion, PFD and Collagen-PVP decrease inflammation, fibrosis, TGFβ-1 expression, and CD in the posttracheoplasty rats' scar.


Adam meets Eph: an ADAM substrate recognition module acts as a molecular switch for ephrin cleavage in trans.
The Eph family of receptor tyrosine kinases and their ephrin ligands are mediators of cell-cell communication. Cleavage of ephrin-A2 by the ADAM10 membrane metalloprotease enables contact repulsion between Eph- and ephrin-expressing cells. How ADAM10 interacts with ephrins in a regulated manner to cleave only Eph bound ephrin molecules remains unclear. The structure of ADAM10 disintegrin and cysteine-rich domains and the functional studies presented here define an essential substrate-recognition module for functional interaction of ADAM10 with the ephrin-A5/EphA3 complex. While ADAM10 constitutively associates with EphA3, the formation of a functional EphA3/ephrin-A5 complex creates a new molecular recognition motif for the ADAM10 cysteine-rich domain that positions the proteinase domain for effective ephrin-A5 cleavage. Surprisingly, the cleavage occurs in trans, with ADAM10 and its substrate being on the membranes of opposing cells. Our data suggest a simple mechanism for regulating ADAM10-mediated ephrin proteolysis, which ensures that only Eph bound ephrins are recognized and cleaved.


Better clinical outcome with direct oral anticoagulants in hospitalized heart failure patients with atrial fibrillation.
Atrial fibrillation (AF) is common in patients with heart failure and is associated with higher mortality. Although previous studies have reported that direct oral anticoagulants (DOACs) reduce the risk of cardiovascular events in out-patients with AF, it remains unclear whether DOACs reduce mortality in hospitalized heart failure (HHF) patients with AF. Therefore, we examined the impact of DOACs on mortality in this group of patients. Consecutive 497 HHF patients with AF were retrospectively registered and divided into three groups on the basis of the presence of anticoagulant therapy: non-anticoagulant group (Non, n = 90), Vit K antagonists (VKAs) group (n = 257) and DOACs group (n = 150). We followed up all the patients for mortality. In the Kaplan-Meier analysis (mean follow-up of 1093 days), all-cause mortality was significantly lower in the VKAs and DOACs groups than in the Non group (31.1% and 15.3% vs. 43.3%, log-rank P < 0.001). In the multivariable Cox proportional hazard analysis after adjusting for other potential confounding factors, usage of DOACs and VKAs were independently associated with lower mortality in HHF patients AF (DOACs, HR 0.356, P = 0.001; VKAs, HR 0.472, P = 0.002). Furthermore, the propensity-matched 1:1 cohort was assessed based on the propensity score (DOACs, n = 114 and VKAs, n = 114). All-cause mortality was significantly lower in the DOACs group than in the VKAs group in the post-matched cohort (12.3% vs. 35.1%, log-rank P = 0.038). In the Cox proportional hazard analysis, the use of DOACs was associated with lower mortality in the post-matched cohort (HR 0.526, P = 0.041). Appropriate use of anticoagulants in HHF patients with AF is important, and DOACs potentially improve all-cause mortality in such patients.


Right ventricular hypertrophy in systemic hypertension: an updated review of clinical studies.
Experimental and clinical evidence supports the view that right ventricular hypertrophy (RVH) may parallel left ventricular hypertrophy in systemic hypertension; a comprehensive analysis of this issue, however, is lacking. Thus, we analyzed the literature in order to provide an updated information on the right ventricular structural changes associated to systemic hypertension. A literature search using the key words 'right ventricle' 'right ventricular hypertrophy', 'biventricular hypertrophy' 'right and left ventricular hypertrophy'. 'hypertension', 'echocardiography' was performed in order to identify relevant articles. Full articles published in English language in the last three decades reporting studies in adult hypertensive individuals were considered. A total of 13 studies, including 1290 untreated (45%) and treated hypertensive patients and 259 normotensive controls, were considered. Overall, in hypertensive individuals right ventricular wall was thicker than in normotensive counterparts (standardized difference 1.3 mm, P < 0.001). RVH prevalence consistently varied among studies (17.0-80.0%) with an average of 28.6% in the pooled population. This was also the case for LVH prevalence rates (9.0-100%) with an average value of 30.6%. Clinical studies consistently indicate that RVH is a common cardiac phenotype in systemic hypertension. As this finding is based on a limited number of cross-sectional studies including small population samples, further investigations are needed to determine the clinical utility and prognostic value of this phenotype in clinical practice.


Breakthrough curves for toluene adsorption on different types of activated carbon fibers: application in respiratory protection.
Activated carbon fibers (ACF) are considered viable alternative adsorbent materials in respirators because of their larger surface area, lighter weight, and fabric form. The purpose of this study was to characterize the breakthrough curves of toluene for different types of commercially available ACFs to understand their potential service lives in respirators. Two forms of ACF, cloth (AC) and felt (AF), with three surface areas each were tested. ACFs were challenged with six toluene concentrations (50-500 p.p.m.) at constant air temperature (23°C), relative humidity (50%), and air flow (16 l min-1) at different bed depths. Breakthrough data were obtained using continuous monitoring by gas chromatography using a gas sampling valve. The ACF specific surface areas were measured by an automatic physisorption analyzer. Results showed unique shapes of breakthrough curves for each ACF form: AC demonstrated a gradual increase in breakthrough concentration, whereas AF showed abrupt increase in concentration from the breakpoint, which was attributed to the difference in fiber density between the forms. AF has steeper breakthrough curves compared with AC with similar specific surface area. AC exhibits higher 10% breakthrough times for a given bed depth due to higher mass per bed depth compared with AF, indicating more adsorption per bed depth with AC. ACF in respirators may be appropriate for use as protection in environments with toluene concentration at the Occupational Safety and Health Administration Permissible Exposure Limit, or during emergency escape for higher toluene concentrations. ACF has shown great potential for application in respiratory protection against toluene and in the development of thinner, lighter, and more efficient respirators.


Rare Earth doped nanoparticles in imaging and PDT.
Nanoparticles doped with rare earth ions for biomedical imaging and infrared photodynamic therapy (IRPDT) have been synthesized, characterized, and compared. Specifically, these nanoparticles utilize two primary modalities: near infrared excitation and emission for imaging, and near infrared upconversion for photodynamic therapy. These nanoparticles are optimized for both their infrared emission and upconversion energy transfer to a photoactive agent conjugated to the surface. Finally, these nanoparticles are tested for toxicity, imaged in cells using the near infrared emission pathway, and used for selective killing of cells through the upconversion driven IRPDT.


Structure-constrained endomorphin analogs display differential antinociceptive mechanisms in mice after spinal administration.
We previously reported a series of novel endomorphin analogs with unnatural amino acid modifications. These analogs display good binding affinity and functional activity toward the μ opioid receptor (MOP). In the present study, we further investigated the spinal antinociceptive activity of these compounds. The analogs were potent in several nociceptive models. Opioid antagonists and antibodies against several endogenous opioid peptides were used to determine the mechanisms of action of these peptides. Intrathecal pretreatment with naloxone and β-funaltrexamine (β-FNA) effectively inhibited analog-induced analgesia, demonstrating that activity of the analogs is regulated primarily through MOP. Antinociception induced by analog 2 through 4 was not reversed by δ opioid receptor (DOP) or κ opioid receptor (KOP) antagonist; antibodies against dynorphin-A (1-17), dynorphin-B (1-13), and Leu5/Met5-enkephalin had no impact on the antinociceptive effects of these analogs. In contrast, antinociceptive effects induced by a spinal injection of the fluorine substituted analog 1 were significantly reversed by KOP antagonism. Furthermore, intrathecal pretreatment with antibodies against dynorphin-B (1-13) attenuated the antinociceptive effect of analog 1. These results indicate that the antinociceptive activity exerted by intrathecally-administered analog 1 is mediated, in part, through KOP with increased release of dynorphin-B (1-13). The chemical modifications used in the present study may serve as a useful tool to gain insight into the mechanisms of endomorphins activity.


Monitoring congenital rubella embryopathy.
Currently, all developed countries include rubella vaccination in their immunization programs, targeting the complete elimination of congenital rubella syndrome (CRS). In the underdeveloped world, where this severely disabling condition still exists, only a few countries have implemented vaccination policies, and almost no data on their effectiveness or on prevalence rates are available. The aims of the present work were to search for the best phenotype to be used as a sentinel for CRS in a large series of malformed newborns and to propose a CRS surveillance system, based only on clinical data. A total of 43 infants diagnosed as having CRS were obtained from 19,184 multimalformed infants, ascertained by the Latin-American Collaborative Study of Congenital Malformations, World Health Organization (WHO) Collaborating Centre for the Prevention of Birth Defects (ECLAMC), over 3,883,165 consecutive births, between 1982 and 2003. They were distributed by country and the most frequent birth defects were identified. From the 19,184 multimalformed infants, all cases presenting the birth defects identified were selected. The sensitivity, specificity, and likelihood ratio (LR) in detecting CRS were determined for these birth defects, alone and in combination. The sample size of multimalformed infants required to detect different levels of increase in the rate of CRS was determined for three sentinel phenotypes. The rate of CRS was highest in Brazil. Based on the best possible combination of sensitivity, specificity, and LR, the dyad comprising eye anomalies and congenital heart defects was shown to be the most appropriate sentinel, with the lowest sample size required, to detect CRS in neonates. A surveillance system for CRS, based on clinical data in newborns, is being proposed, in an attempt to monitor ongoing vaccination policies, aimed at eliminating CRS in developing countries.


[Gene expression of gonadotropin releasing hormone in the rat brain--an in situ hybridization technic using an oligoRNA probe].
We studied the localization of GnRH cell bodies in the rat brain by using an in situ hybridization technique. We used radiolabeled single stranded oligoRNA probe (62-mer) complementary to the part of rat GnRH mRNA which included the coding sequence for the decapeptide. Only a single hybridizing band was observed on Northern blotting of mRNA extracts of the rat brain. This probe detects GnRH mRNA in individual cells in sections of the rat brain. In situ hybridization revealed the presence of GnRH cell bodies in the preoptic area and diagonal bands of Broca. We found that the oligoRNA probe was useful because it was easy to prepare and accessible to cellular mRNA.


Brain-aging related protein expression and imaging characteristics of mice exposed to chronic hypoxia at high altitude.
To determine changes in protein expression related to brain aging and imaging features in mice after chronic hypoxia exposure at high altitude. A total of 24 healthy 4-week-old mice were randomly divided into high altitude hypoxia (HH) and plain control (PC) groups (n = 8 per group). HH mice were transported from Xi'an (450 m above sea level) to Maduo (4,300 m above sea level) while PC mice were raised in Xi'an. After 6 months, 7.0T magnetic resonance imaging (MRI) was performed. All mice completed T2-weighted imaging (T2WI), diffusion tensor imaging (DTI), resting-state functional MRI (rs-fMRI), arterial spin labeling (ASL), and magnetic resonance angiography (MRA) examinations. Next, brain slices were prepared and Nissl staining was used to observe morphological changes in neurons. Ultrastructural changes in neurons were observed by transmission electron microscopy. Expression changes of Caspase-3, klotho, P16, P21, and P53 at the gene and protein levels were detected by real-time PCR (RT-PCR) and Western blot. The number of neuronal Nissl bodies in the hippocampus and frontal cortex was significantly decreased in the HH group compared to the PC group. Some hippocampal and frontal cortical neurons were apoptotic, the nuclei were wrinkled, chromatin was aggregated, and most mitochondria were mildly swollen (crista lysis, fracture). Compared with the PC group, the HH group showed elevated expression of caspase-3 mRNA, P16 mRNA, P21 mRNA, and P53 mRNA in the hippocampus and frontal cortex. Expression of Klotho mRNA in the frontal cortex was also significantly decreased. Western blot results showed that caspase-3 protein expression in the hippocampus and frontal cortex of the HH group was increased compared with the PC group. Moreover, there was decreased Klotho protein expression and significantly increased P-P53 protein expression. Compared with the PC group, expression of P16 protein in the frontal cortex of the HH group was increased and the gray matter (GM) volume in the left visceral area, left caudate nucleus, and left piriform cortex was decreased. Furthermore, the amplitude of low frequency fluctuation was decreased in the left posterior nongranular insular lobe, right small cell reticular nucleus, left flocculus, left accessory flocculus, and left primary auditory area, but increased in the GM layer of the left superior colliculus. Regional homogeneity was decreased in the left and right olfactory regions, but increased in the left bed nucleus. After exposure to high altitude, functional connectivity (FC) between the bilateral caudate nucleus and thalamus, corpus callosum, cingulate gyrus, anterior limbic cortex, globus pallidus, and hippocampus was weakened. FC between the right caudate nucleus and hypothalamus and entorhinal cortex was also weakened. The fractional anisotropy value of the left hippocampus was decreased in the HH group. Compared with the PC group, the HH group showed significantly increased inner diameters of the bilateral common carotid artery and left internal carotid artery. The cerebral blood flow values of the bilateral cortex and bilateral hippocampus in the HH group did not change significantly. Taken together, our findings show that chronic hypoxia exposure at high altitude may promote neuronal apoptosis and abnormal expression of related proteins, changing the structure and function of brain. These changes may contribute to brain aging.


Ehrlich ascites tumor cells produce a transforming growth factor-beta (TGFbeta)-like activity but lack receptors with TGFbeta-binding capacity.
Ehrlich ascites tumor cells incorporate [methyl-3H]thymidine into DNA independently of exogenous growth factors or fetal calf serum. Using an acid/ethanol extraction procedure we have obtained from these tumor cells a fraction that induces both the proliferation and the formation of cell foci by Swiss 3T3 mouse fibroblasts in the presence of insulin; inhibits the proliferation of Mv1Lu mink lung epithelial cells; and stimulates the growth of NRK rat kidney fibroblasts in soft-agar in the presence of epidermal growth factor. An antibody against transforming growth factor-beta (TGFbeta) prevents both the tumor extract-induced proliferation of Swiss 3T3 fibroblasts and the tumor extract-induced proliferative arrest of Mv1Lu cells. The tumor cells secrete a TGF beta-like activity to the extracellular medium in a partially-activated form. However, authentic TGFbeta does not affect their proliferation, and no TGFbeta receptors were detected using [125I]TGFbeta as a ligand. Therefore, the absence of TGFbeta receptors with ligand-binding capacity in these tumor cells may bypass the negative control that this factor exerts on the proliferation of their normal cell counterparts.


Prevalence, awareness, treatment and control of hypertension in an elderly population in Greece.
Hypertension is a major public health problem and its prevalence increases with age. Despite an aging population, only a limited number of population-based studies, and fewer Greek studies, have focused exclusively on the elderly. The objective of this study was to investigate the prevalence, awareness, treatment and control of hypertension in the rural residents of Paliouri village in Greece who were aged 65 years and older. In total there were 171 participants (response rate 89.5%). Six blood pressure (BP) measurements were obtained in two visits and hypertension was defined as a mean systolic BP >or= 140 mmHg and/or mean diastolic BP >or= 90 mmHg, and/or current use of an antihypertensive medication. The prevalence of hypertension was 89%. Of the 137 hypertensive patients, 89.8% were aware of their disease, 89.1% were receiving treatment, and the hypertension of 32.8% was controlled. Awareness of hypertension was significantly higher among those with a high frequency of BP measurements, a history of coronary or other chronic disease, lower education, and those living with a spouse. The findings indicate that hypertension is highly prevalent in this Greek elderly population. Similarly high is the awareness and treatment of hypertension. However, the control rates were relatively low, suggesting that the local health system requires a greater orientation to the prevention and control of hypertension.


Cardioprotective effect of ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy in experimental animals.
The objective of this study was to evaluate the cardioprotective effect of herbal bioactive compound ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy, in experimental animals. Ammonium glycyrrhizinate (50, 100, 200 mg/kg, p.o.) was administered for four weeks in albino rats. Cardiomyopathy was induced with a dose of 2.5 mg/kg i.p. of doxorubicin on 1(th), 7(th), 14(th), 21(th), 28(th) day in the experimental animals. At the end of the experiment, on 29(th) day, serum and heart tissues were collected and hemodynamic, biochemical and histopathological studies were carried out. Administration of doxorubicin in normal rats showed significant (P < 0.001) changes in body weight, feed intake, urine output, hemodynamic parameters like (blood pressure, heart rate, cardiac output) and in lipid profile (cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, very low density lipoprotein) indicating cardiomyopathy symptoms. Animals treated with ammonium glycyrrhizinate significantly (P < 0.05) decreased triglyceride, cholesterol, low density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels. Moreover, high density lipoprotein (HDL) levels increased in rats treated with ammonium glycyrrhizinate as compared with the normal group. Ammonium glycyrrhizinate is effective in controlling serum lipid profile and cardiac complications in experimentally induced cardiomyopathy in animals.


Peculiarities of digestion in senescence.
It is established, that involutional changes of digestive tract limits digestion in old patients. In the age digestive capacities mostly are sufficient, but there is a great tendency to maldigestion. The decrease of functional capacity doesn't show the clinical features of a manifest digestive insufficiency in old healthy probands. But it was shown that the digestive efficiency is exhausted by hyper-alimentation more quickly in elderly. The pancreas of elderly is able to digest small meals taken frequently.


Practical guide for selection of 1 H qNMR acquisition and processing parameters confirmed by automated spectra evaluation.
In our recent paper, a new technique for automated spectra integration and quality control of the acquired results in qNMR was developed and validated (Monakhova & Diehl, Magn. Res. Chem. 2017, doi: 10.1002/mrc.4591). The present study is focused on the influence of acquisition and postacquisition parameters on the developed automated routine in particular, and on the quantitative NMR (qNMR) results in general, which has not been undertaken previously in a systematic and automated manner. Results are presented for a number of model mixtures and authentic pharmaceutical products measured on 500- and 600-MHz NMR spectrometers. The influence of the most important acquisition (spectral width, transmitter [frequency] offset, number of scans, and time domain) and processing (size of real spectrum, deconvolution, Gaussian window multiplication, and line broadening) parameters for qNMR was automatically investigated. Moderate modification of the majority of the investigated parameters from default instrument settings within evaluated ranges does not significantly affect the trueness and precision of the qNMR. Lite Gaussian window multiplication resulted in accuracy improvement of the qNMR output and is recommended for routine measurements. In general, given that the acquisition and processing parameters were selected based on the presented guidelines, automated qNMR analysis can be employed for reproducible high-precision concentration measurements in practice.


A population genetic model for the maintenance of R2 retrotransposons in rRNA gene loci.
R2 retrotransposable elements exclusively insert into the tandemly repeated rRNA genes, the rDNA loci, of their animal hosts. R2 elements form stable long-term associations with their host, in which all individuals in a population contain many potentially active copies, but only a fraction of these individuals show active R2 retrotransposition. Previous studies have found that R2 RNA transcripts are processed from a 28S co-transcript and that the likelihood of R2-inserted units being transcribed is dependent upon their distribution within the rDNA locus. Here we analyze the rDNA locus and R2 elements from nearly 100 R2-active and R2-inactive individuals from natural populations of Drosophila simulans. Along with previous findings concerning the structure and expression of the rDNA loci, these data were incorporated into computer simulations to model the crossover events that give rise to the concerted evolution of the rRNA genes. The simulations that best reproduce the population data assume that only about 40 rDNA units out of the over 200 total units are actively transcribed and that these transcribed units are clustered in a single region of the locus. In the model, the host establishes this transcription domain at each generation in the region with the fewest R2 insertions. Only if the host cannot avoid R2 insertions within this 40-unit domain are R2 elements active in that generation. The simulations also require that most crossover events in the locus occur in the transcription domain in order to explain the empirical observation that R2 elements are seldom duplicated by crossover events. Thus the key to the long-term stability of R2 elements is the stochastic nature of the crossover events within the rDNA locus, and the inevitable expansions and contractions that introduce and remove R2-inserted units from the transcriptionally active domain.


Signal denoising and ultrasonic flaw detection via overcomplete and sparse representations.
Sparse signal representations from overcomplete dictionaries are the most recent technique in the signal processing community. Applications of this technique extend into many fields. In this paper, this technique is utilized to cope with ultrasonic flaw detection and noise suppression problem. In particular, a noisy ultrasonic signal is decomposed into sparse representations using a sparse Bayesian learning algorithm and an overcomplete dictionary customized from a Gabor dictionary by incorporating some a priori information of the transducer used. Nonlinear postprocessing including thresholding and pruning is then applied to the decomposed coefficients to reduce the noise contribution and extract the flaw information. Because of the high compact essence of sparse representations, flaw echoes are packed into a few significant coefficients, and noise energy is likely scattered all over the dictionary atoms, generating insignificant coefficients. This property greatly increases the efficiency of the pruning and thresholding operations and is extremely useful for detecting flaw echoes embedded in background noise. The performance of the proposed approach is verified experimentally and compared with the wavelet transform signal processor. Experimental results to detect ultrasonic flaw echoes contaminated by white Gaussian additive noise or correlated noise are presented in the paper.


Analysis of family resemblance. V. Height and weight in northeastern Brazil.
Sib correlations for height and weight decrease with absolute age difference. Parent-child correlations increase with age of child, with greater resemblance to the mother than the father. Estimates of the relative variance due to common environment are greater, and heritability estimates less, than for earlier studies. Heritability is less for adults than children. There is no significant major locus for height or weight.


HIV-1 gp120 Upregulates Brain-Derived Neurotrophic Factor (BDNF) Expression in BV2 Cells via the Wnt/β-Catenin Signaling Pathway.
HIV-1 gp120 plays a critical role in the pathogenesis of HIV-associated pain, but the underlying molecular mechanisms are incompletely understood. This study aims to determine the effect and possible mechanism of HIV-1 gp120 on BDNF expression in BV2 cells (a murine-derived microglial cell line). We observed that gp120 (10 ng/ml) activated BV2 cells in cultures and upregulated proBDNF/mBDNF. Furthermore, gp120-treated BV2 also accumulated Wnt3a and β-catenin, suggesting the activation of the Wnt/β-catenin pathway. We demonstrated that activation of the pathway by Wnt3a upregulated BDNF expression. In contrast, inhibition of the Wnt/β-catenin pathway by either DKK1 or IWR-1 attenuated BDNF upregulation induced by gp120 or Wnt3a. These findings collectively suggest that gp120 stimulates BDNF expression in BV2 cells via the Wnt/β-catenin signaling pathway.


Prescriptive appropriateness using inhalant and food allergen panels: a comparison between General Practitioners' and Allergists' prescription in Genoa (Italy).
Background. Prescriptive appropriateness is an actual claim in healthcare, and it also concerns in vitro tests used in the allergy work-up, such as the serum allergen-specific IgE (sIgE) assay. In the Liguria Region, two panels were defined (for inhaled and food allergens) including 12 allergens. Their composition changed over time. Objectives. The aims of the present retrospective study were: i) to evaluate the percentage of positive tests, and ii) to compare the findings of sIgE assay on the basis of the general practictioners' (GPs) or specialist' prescription, considering both the old panels and the new panels. Methods. This retrospective study considered a population of adult patients, which consisted of 2368 subjects (68% females; mean age 50 years; age range: 10-103 years). Serum sIgE were measured by ImmunoCap system. Results. The percentages of positive tests were very low for food allergens and low for inhaled ones (ranging between 5% to 35%). There was change of prevalent prescriptor with new panels. Conclusions. This study underlines the relevance of prescriptive appropriateness in the allergy work-up. The sIgE assay should be limited to those allergens that have a clinical relevance, based on clinical history.


[Enhancing Soft-tissue Reattachment with Artificial Mesh in Joint Endoprosthetic Reconstruction for Bone Tumors].
Objective: To investigate the operative method and clinical application of the BARD(®) Mesh in enhancing joint stability and function of endoprosthetic reconstruction for bone tumors. Methods: From Jan 2013 to Jun 2015, the clinical data of 51 patients aged (44.75±23.18) years underwent wide resection of tumor and endoprosthetic reconstruction using the BARD(®) Mesh were collected. Among them, 27 were male and 24 were female. The surgical treatments received by these patients included 5 shoulder arthroplasties, 12 elbow replacements, 12 hip replacements and 32 knee replacements (including 24 femoral tumors and 8 tibial tumors). According to the pathologic type, there were 12 metastatic tumors, 20 osteosarcomas, 7 chondrosarcomas, 5 malignant fibrous histiocytomas, 4 giant cell tumors of bone, 1 Ewing sarcoma, 1 leiomyosarcoma and 1 pigmented villonodular synovitis (pvns). These patients received extensive tumor resection, tumorous prosthesis replacement, preserved articular capsule and muscles repair with artificial mesh and endoprosthesis wrapping. The curative effect including joints range of motion and Musculoskeletal Tumour Society Scores (MSTS) were evaluated. Results: The median follow-up time was (19.75±8.17) months. The drainages were removed out on an average of 4 days after operation. The postoperative complications included 2 superficial incision infection, 1 deep incision infection and 1 osteofascial compartment syndrome, infection or dislocation of prosthesis wasn't observed. The mean active flexion of shoulder joint after replacement was (34.00±10.84)°, mean active abduction was (20.00±9.35)° and the mean MSTS was 19.80±9.54. The superior rate of shoulder flexion function was 0. The mean active flexion of elbow joint after replacement was (75.00±7.07)°, mean active abduction was (-5.00±7.07)° and the mean MSTS was 25.00±2.83. The superior rate of elbow flexion function was 50.0% (1/2). The mean active flexion of hip joint after replacement was (86.67±20.60)°, mean active abduction was (2.08±4.98)° and the mean MSTS was 25.42±1.78. The superior rate of hip flexion function was 83.3% (10/12). The mean active flexion of knee joint after replacement was (89.69±22.39)°, mean active abduction was (-0.63±1.68)° and the mean MSTS was 23.31±2.09. The superior rate of knee flexion function was 50.0%(16/32). Among them, the superior rate of femoral flexion function was 66.7% (16/24), the superior rate of tibial flexion function was 0. All of patients were satisfied with the curative effect of operation at the end of follow-up time. Conclusions: The BARD(®) Mesh may enhance the attachment of soft-tissue to endoprosthesis, improve the joint stability, decrease the endoprosthetic infection and dislocation, facilitate the attachment of tendon to endoprosthesis and recover the muscular motivation after endoprosthetic reconstruction. This plays an important role in joint stability and motivation reconstruction of soft-tissue impairment, effectively prevents surgical complications.


Anatomical characterisation of three different psychosurgical targets in the subthalamic area: from the basal ganglia to the limbic system.
Effective neural stimulation for the treatment of severe psychiatric disorders needs accurate characterisation of surgical targets. This is especially true for the medial subthalamic region (MSR) which contains three targets: the anteromedial STN for obsessive compulsive disorder (OCD), the medial forebrain bundle (MFB) for depression and OCD, and the "Sano triangle" for pathological aggressiveness. Blocks containing the subthalamic area were obtained from two human brains. After obtaining 11.7-Tesla MRI, blocks were cut in regular sections for immunohistochemistry. Fluorescent in situ hybridisation was performed on the macaque MSR. Electron microscopic observation for synaptic specialisation was performed on human and macaque subthalamic fresh samples. Images of human brain sections were reconstructed in a cryoblock which was registered on the MRI and histological slices were then registered. The STN contains glutamatergic and fewer GABAergic neurons and has no strict boundary with the adjacent MSR. The anteromedial STN has abundant dopaminergic and serotoninergic innervation with very sparse dopaminergic neurons. The MFB is composed of dense anterior dopaminergic and posterior serotoninergic fibres, and fewer cholinergic and glutamatergic fibres. Medially, the Sano triangle presumably contains orexinergic terminals from the hypothalamus, and neurons with strong nuclear oestrogen receptor-alpha staining with a decreased anteroposterior and mediolateral gradient of staining. These findings provide new insight regarding MSR cells and their fibre specialisation, forming a transition zone between the basal ganglia and the limbic systems. Our 3D reconstruction enabled us to visualize the main histological features of the three targets which should enable better targeting and understanding of neuromodulatory stimulation results in severe psychiatric conditions.


[Homeless women with psychiatric disorders -- a field study].
The aim of the study was to investigate the psychosocial situation of all homeless women in a southern German university town. The scope, structure and whereabouts of the target group were initially unknown. 17 of 22 homeless women identified agreed to participate in an extensive study including the Structured Clinical Interview for DSM-IV (SKID-I). The prevalence of diagnosed psychiatric disorders was 71 %; the leading disorder was substance abuse/dependence (43 %), followed by anxiety disorders (35 %) and schizophrenia (12 %). Multiple diagnoses were made in 35 % of the women. Striking features were the often early onset of homelessness and the reticence in seeking help. The flight from violence was a crucial precipitant of the loss of the home and should also be discussed in the context of the development of the psychiatric disorders. This study is the first involving women outside major cities. Precipitant situation, need for help and help-seeking pattern appear to deviate markedly from those of homeless men. The results should be verified in further gender-specific studies.


Impact of the pay-for-performance program on lower extremity amputations in patients with diabetes in Taiwan.
Patients with diabetes are at a high risk of lower extremity amputations and may have a reduced life expectancy. Taiwan has implemented a diabetes pay-for-performance (P4P) program providing team care to improve the control of disease and avoid subsequent complications. Few studies investigated the effects of adopting a nationalized policy to decrease amputation risk in diabetes previously. Our study aimed to analyze the impact of the P4P programs on the incidence of lower extremity amputations in Taiwanese patients with diabetes.This was a population-based cohort study using the Taiwan National Health Insurance Research Database (which provided coverage for 98% of the total population in Taiwan) from 1998 to 2007. Patients with diabetes were identified based on the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) diagnostic codes. We linked procedure codes to inpatient claims to identify patients hospitalized for nontraumatic lower extremity amputations.A total of 9738 patients with diabetes with amputations were enrolled (mean age ± standard deviation: 64.4 ± 14.5 years; men: 63.9%). The incidence of nontraumatic diabetic lower extremity amputations decreased over the time period studied (3.79-2.27 per 1000 persons with diabetes). Based on the Cox proportional hazard regression model, male sex (hazard ratio: 1.83, 95% confidence interval [CI] 1.76-1.92), older age, and low socioeconomic status significantly interact with diabetes with respect to the risks of amputation. Patients who did not join the P4P program for diabetes care had a 3.46-fold higher risk of amputation compared with those who joined (95% CI 3.19-3.76).The amputation rate in Taiwanese diabetic patients decreased over the time period observed. Diabetes in patients with low socioeconomic status is associated with an increased risk of amputations. Our findings suggested that in addition to medical interventions and self-management educations, formulate and implement of medical policies, such as P4P program, might have a significant effect on decreasing the diabetes-related amputation rate.


Expanding the role of PAs in the treatment of severe and persistent mental illness.
Patients with severe and persistent mental illness often face limited access to psychiatric and primary care. This exploratory qualitative analysis studied the effect of introducing a PA into a comprehensive outpatient psychiatric team. Early results suggest this model improves the quality and access to primary and psychiatric care.


Considerations in the Development of a Sound Tolerance Interview and Questionnaire Instrument.
Most clinicians approach the objective fitting of hearing aids with three goals in mind: audibility, comfort, and tolerance. When these three amplification goals have been met, the hearing aid user is more likely to adapt to and perceive benefit from hearing aid use. However, problems related to the loudness of sounds and reduced sound tolerance, which may or may not be reported by the aided user, can adversely impact adaptation to amplification and the individual's quality of life. Although there are several standardized questionnaires available to evaluate hearing aid benefit and satisfaction, there is no standardized questionnaire or interview tool for evaluating reduced sound tolerance and the related impact on hearing aid use. We describe a 36-item tool, the Sound Tolerance Questionnaire (STQ), consisting of six sections, including experience with hearing aids, sound sensitivity/intolerance, medical and noise exposure histories, coexisting tinnitus problems, and a final question to differentiate the primary and secondary problems related to sound intolerance, tinnitus, and hearing loss. In its current format as a research tool, the STQ was sensitive in pinpointing vague sound tolerance complaints not reported by the study participants in eligibility screening by Formby et al. A refined version of the STQ, the Sound Tolerance Interview and Questionnaire Instrument (STIQI), structured as a two-part tool, is presented in the appendix for prospective clinical use. The STIQI has potential utility to delineate factors contributing to loudness complaints and/or reduced sound tolerance in individuals considering hearing aid use, as well as those who have been unsuccessful hearing aid users secondary to loudness complaints or sound intolerance. The STIQI, when validated and refined, also may hold promise for predicting hearing aid benefit and/or assessing treatment-related change over time of hearing aid use or interventions designed to remediate problems of loudness and/or sound intolerance among hearing aid candidates or users.


Isolation of the major serine protease inhibitor from the 5-day serum-free conditioned medium of human embryonic lung cells and demonstration that it is fetuin.
Human embryonic lung (HuEL) cells in culture produce large amounts of the enzyme, plasminogen activator, and thus generate substantial amounts of active plasmin. Despite the presence of plasmin, however, HuEL cells grow in ordered, flattened, adherent sheets. It seemed of interest to characterize protease inhibitors that might be present in HuEL cultures and which might account for this apparent contradiction. This paper reports the isolation and purification of the major serine protease inhibitor in 5-day serum-free conditioned medium (CM) from HuEL cells, and the purification of an identical molecule from fetal bovine serum (FBS). Both the CM-derived inhibitor and the FBS-derived inhibitor are identical with fetuin, the major glycoprotein of FBS. The CM-derived inhibitor is apparently derived from the FBS used to supplement the growth medium of HuEL cells between serum-free CM collection periods. It is not labeled metabolically with 3H-leucine. Its electrophoretic behavior is indistinguishable from that of standard fetuin in SDS-PAGE, non-SDS basic pH,PAGE, and isoelectric focusing. The CM-derived inhibitor and standard fetuin inhibit trypsin and plasmin with similar efficiencies, but neither inhibits chymotrypsin, pancreatic elastase, or plasminogen activator. They are immunologically indistinguishable. The suggestion is made that fetuin, and possibly other protease inhibitors present in HuEL cell cultures, may be concentrated locally by HuEL cells and gradually released back into the medium in the absence of serum. These molecules may serve to protect HuEL cells against proteases they generate.


Serum amyloid A and interleukin -1β facilitate LDL transcytosis across endothelial cells and atherosclerosis via NF-κB/caveolin-1/cavin-1 pathway.
Inflammatory molecules play important roles in atherosclerosis. We aimed to illustrate the roles of serum amyloid A (SAA), and interleukin (IL)-1β in low density lipoproteins (LDL) transcytosis and atherosclerosis. Effects of SAA and IL-1β on transcytosis of LDL were measured by an in vitro LDL transcytosis model. NF-κB/caveolin-1/cavin-1 pathway activation was investigated by Western blots and ELISA. Effects of SAA and IL-1β on the retention of LDL in aorta of C57BL/6J mice were detected by IVIS spectrum. Effects of SAA and IL-1β on atherosclerosis in Apoe-/- mice were examined by Oil Red O staining. SAA and IL-1β stimulated LDL transcytosis across endothelial cells (ECs), which was accompanied by an increase in LDL uptake by ECs. SAA and IL-1β enhanced the activity of nuclear factor (NF)-κB, consequently facilitating an up-regulation of proteins involved in caveolae formation, including caveolin-1 and cavin-1, along with an assembly of NLRP3 inflammasome. Furthermore, SAA- and IL-1β-induced effects were blocked by NF-κB subunit p65 siRNA. Meanwhile, SAA- and IL-1β-induced LDL transcytosis were effectively blocked by caveolin-1 siRNA or cavin-1 siRNA. Interestingly, SAA and IL-1β facilitated LDL entering into the aorta of C57BL/6J mice. In Apoe-/- mice, SAA and IL-1β increased the areas of lipid-rich atherosclerotic lesions in the both ascending and root of aorta. Furthermore, a significant increase in the NLRP3 inflammasome, accompanied by accumulation of cavin-1 and caveolin-1, was observed in the aortic endothelium of Apoe-/- mice. SAA and IL-1β accelerated LDL transcytosis via the NF-κB/caveolin-1/cavin-1 axis.


Kidney progression project (KiPP): Protocol for a longitudinal cohort study of progression in chronic kidney disease of unknown etiology in Sri Lanka.
Over the last two decades, a global epidemic of chronic kidney disease of unknown etiology (CKDu) has emerged in rural, arid, agricultural, lowland areas. Endemic regions have reported 15 to 20% prevalence among residents aged 30-60 years. CKDu is a progressive and irreversible disease resulting in renal failure and death in the absence of dialysis or a kidney transplant. While much of the research has focused on identifying etiology, this project seeks to ascertain factors associated with the rapidity of kidney disease progression in one of Sri Lanka's CKDu endemic areas. A sample of 296 male and female residents aged 21 to 65 with moderate CKD, as measured by their serum creatinine level, and a clinical diagnosis of CKDu are followed using quarterly serum testing to track the rate of progression. A baseline survey administered to the entire sample addresses potential risk factors, supplemented by a short survey focusing on changes through time. Concurrently water, soil and air are tested at the local and household levels. The study is the first to foster a multi-disciplinary approach that focuses on disease progression, identifying behavioural and exposure risk factors for rapid kidney function decline, in this progressively fatal disease.


3-Phosphoglycerate is an allosteric activator of pyruvate kinase from the hyperthermophilic archaeon Pyrobaculum aerophilum.
Pyruvate kinase (PK) is a highly regulated enzyme that catalyzes the final step of glycolysis. PK from the hyperthermophilic archaeon Pyrobaculum aerophilum (PaPK) is distinguished from most PK enzymes of eukarya and bacteria by not responding to any known allosteric effectors and apparently exhibiting only cooperative regulation. We determined the crystal structure of PaPK to 2.2 Å resolution and, in a manner consistent with the lack of a response to conventional effectors, observed that the canonical allosteric site is occluded by a tyrosine. Unexpectedly, though, a bound sulfate was observed at a position equivalent to the 6'-phosphate of sugar effectors, suggesting an allosteric site, but for an unknown effector and sharing only the phosphate position. A search of three-carbon intermediates of glycolysis revealed 3-phosphoglycerate (3PG) as a potent allosteric activator of PaPK. The response was abolished by mutation of residues that contact the sulfate and of an arginine proposed to interact with the 3PG carboxylate group. Regulation of PK by 3PG is consistent with the ancestral glycolysis of hyperthermophilic archaea in which this intermediate is produced by an irreversible enzyme, glyceraldehyde 3-phosphate ferredoxin oxidoreductase. Coordinated regulation within the lower half of glycolysis contrasts sharply with conventional glycolysis in which 3PG is produced reversibly and PK is regulated by fructose 1,6-bisphosphate, the product of phosphofructokinase, an irreversible enzyme in the upper half of the pathway. Regulation of PaPK by a carboxylate molecule rather than a sugar phosphate may reflect a step in the evolution of glycolysis that predates the dominance of sugars in metabolism.


[Prevalence and comorbidity of neuropsychiatric symptoms in Alzheimer's disease].
The objective of this study is to describe the frequency and severity of behavioral and psychological symptoms (BPS) in a group of 125 patients diagnosed of Alzheimer's disease (AD) (DSM-IV-TR and NINCDSADRDA criteria). The evaluation of the BPS was carried out using the Neuropsychiatric Inventory (NPI; Cummings et al., 1994). The sociodemographic and personal background data of the patients were gathered and the dementia stage was established with the Global Deterioration Scale (GDS Reisberg, 1982). A total of 122 patients (98%) presented BPS, with an average of five symptoms per patient. Frequency of presentation was the following: apathy (75%), irritability (66%), depression (60%), agitation (55%), anxiety (54%), aberrant motor activity (47%), delirium (38%), sleeping disorders (36%), disinhibition (29%), eating disorders (28%), hallucinations (20%) and euphoria (4%). These results show the high incidence of BPS in AD patients and point to the necessity and importance of treating these disorders appropriately.


Understanding prosthodontics--where did it all go wrong?
This paper addresses the anatomical and physiological effects of tooth loss on the patient and, further, audits the teaching of prosthodontics over the last 30 years. There is an abundance of evidence to outline changes that occur post-extraction related to morphological changes in the maxilla, the mandible and combined changes in inter-maxillary relations. In addition to the knowledge of patterns of bone loss, we are also now well aware that there is evidence to suggest that planned retention of roots and/or the placement of implants can help reduce post-extraction bone loss. Awareness by clinicians of the anatomical, physiological and psychological effects of edentulousness, is essential if empathy is to be struck with the patient. Equally, patients being rendered edentulous must be made aware of these factors if true informed consent is to be gained. New graduates are now less well equipped to practice complete denture prosthodontics than, say, 10 years ago and this is a consequence of three factors: crowded curricula, diminution in laboratory hours and the time a dental student spends in clinical prosthodontics.


Structure and transcription of the genes encoding the B1015 light-harvesting complex beta and alpha subunits and the photosynthetic reaction center L, M, and cytochrome c subunits from Rhodopseudomonas viridis.
The genes encoding the beta and alpha subunits of the B1015 light-harvesting complex (LHC) and the L, M, and cytochrome c subunits of the photosynthetic reaction center from Rhodopseudomonas viridis are organized in an operon, in analogy to other nonsulfur purple bacteria, named the puf operon. In photoheterotrophically grown cells, two abundant puf operon mRNA species of 3,581 and 621 bases were present. The large transcript encoded the LHC beta, LHC alpha, and reaction center L, M, and cytochrome c polypeptides, whereas the small transcript only coded for the LHC beta and alpha polypeptides. Both transcripts share a common 5' end which is located 115 bases upstream from the initiation codon of the LHC beta gene. Two additional low-level transcripts of 3,718 and 758 bases with 5' ends 254 +/- 3 bases upstream from the LHC beta gene were detected. Analysis of the DNA sequence preceding the different 5' ends revealed DNA elements of striking homology. The 3' ends of the small transcripts were mapped within the alpha-L intercistronic DNA region downstream from a sequence capable of forming a very stable stem-loop when transcribed into RNA. The 3' termini of the large transcripts are located immediately downstream from the region coding the cytochrome c subunit in two areas resembling rho-independent transcription terminators. No open reading frames corresponding to pufQ and pufX from Rhodobacter capsulatus and Rhodobacter sphaeroides were present in the flanking DNA regions of the puf operon. In contrast, an open reading frame ending 191 base pairs upstream from the LHC beta gene showed 50% homology at the amino acid level to the available sequence of the bchA gene from R. capsulatus. The genes coding for the B1015 LHC subunits had C-terminal extensions of 13 (beta) and 10 (alpha) amino acids which were not present in the proteins isolated from intracytoplasmic membranes.


Characterization of Central and Nasal Orbital Adipose Stem Cells and their Neural Differentiation Footprints.
The unique potential of stem cells to restore vision and regenerate damaged ocular cells has led to the increased attraction of researchers and ophthalmologists to ocular regenerative medicine in recent decades. In addition, advantages such as easy access to ocular tissues, non-invasive follow-up, and ocular immunologic privilege have enhanced the desire to develop ocular regenerative medicine. This study aimed to characterize central and nasal orbital adipose stem cells (OASCs) and their neural differentiation potential. The central and nasal orbital adipose tissues extracted during an upper blepharoplasty surgery were explant-cultured in Dulbecco's Modified Eagle Medium (DMEM)/F12 supplemented with 10% fetal bovine serum (FBS). Cells from passage 3 were characterized morphologically by osteogenic and adipogenic differentiation potential and by flow cytometry for expression of mesenchymal (CD73, CD90, and CD105) and hematopoietic (CD34 and CD45) markers. The potential of OASCs for the expression of NGF, PI3K, and MAPK and to induce neurogenesis was assessed by real-time PCR. OASCs were spindle-shaped and positive for adipogenic and osteogenic induction. They were also positive for mesenchymal and negative for hematopoietic markers. They were positive for NGF expression in the absence of any significant alteration in the expression of PI3K and MAPK genes. Nasal OASCs had higher expression of CD90, higher potential for adipogenesis, a higher level of NGF expression under serum-free supplementation, and more potential for neuron-like morphology. We suggested the explant method of culture as an easy and suitable method for the expansion of OASCs. Our findings denote mesenchymal properties of both central and nasal OASCs, while mesenchymal and neural characteristics were expressed stronger in nasal OASCs when compared to central ones. These findings can be added to the literature when cell transplantation is targeted in the treatment of neuro-retinal degenerative disorders.


MicroRNA-199a-3p inhibits tumorigenesis of hepatocellular carcinoma cells by targeting ZHX1/PUMA signal.
MicroRNAs play an important role in cell proliferation, apoptosis, differentiation, and invasion by regulating the expression of various genes. For example, the downregulation of microRNA-199a-3p (miR-199a-3p) that is noted in numerous human malignancies, including hepatocellular carcinoma (HCC), results in a poor prognosis in patients with HCC. This finding suggests that miR-199a-3p overexpression in HCC could provide a new treatment approach. We explored this possibility by examining the effects of miR-199a-3p on the growth and apoptosis of HCC cells in vitro and vivo. The miR-199a-3p signaling pathway was examined using ZHX1 (zinc-fingers and homeoboxes-1) or PUMA (a p53 upregulated modulator of apoptosis) siRNA transfection to determine the effects of miR-199a-3p on growth and apoptosis of HepG2 cells in vitro. A subcutaneously implanted tumor model of HepG2 cells in nude mice was used to assess the effects of miR-199a-3p on the signaling pathway and tumorigenesis development in vivo. miR-199a-3p inhibited growth and induced apoptosis of HepG2 cells in vitro. These effects were accompanied by upregulation of ZHX1 and PUMA. Targeting ZHX1 inhibited upregulation of PUMA after miR-199a-3p transfection. In addition, miR-199a-3p inhibited Bcl2 expression, but increased Bax and cleaved caspase-3 expression. Targeting PUMA or ZHX1 reversed the effect of miR-199a-3p, followed by upregulation of Bcl2 and downregulation of Bax and cleaved caspase-3, respectively. Furthermore, miR-199a-3p inhibited tumorigenesis of xenografts in nude mice. miRNA-199a-3p could effectively prevent primary tumor formation. The ability of this therapy to decrease tumorigenesis may be related toZHX1-dependent PUMA signals.


Extracellular vesicles of bovine small follicular fluid promote ovarian cortical stromal cell proliferation and steroidogenesis.
The aim of this study was to investigate the effects of extracellular vesicles (EVs) on the proliferation and steroid hormone synthesis of bovine ovarian cortical stromal cells in vitro. The release and uptake of EVs are the new mechanisms of cell-to-cell communication. Using reverse transcriptase polymerase chain reaction, enzyme-linked immunosorbent assay, TUNEL and other experiments, we found that EVs in bovine follicular fluid can promote the proliferation and synthesis of androstenedione and progesterone in ovarian cortical stromal cells. Moreover, 100 μg/ml EVs caused the most significant effect. We conclude that EVs at 100 μg/ml can significantly promote the proliferation and synthesis of androstenedione and progesterone in ovarian cortical stromal cells. This research is of great significance for further elucidating the regulatory role of follicular fluid EVs in follicular development and atresia and for research on the interaction of ovarian stromal cells, granulosa cells and oocytes.


[Myeloid hemopoietic growth factors. Therapeutic possibilities and clinical experiences].
Colony stimulating factors (CSF) for white blood cells are formed in the bone-marrow and regulate proliferation and differentiation of the myeloid cells. Several of these hormones are cloned and manufactured by a recombinant technique for clinical use. The granulocyte-macrophage-colony-stimulating factor (GM-CSF) and the granulocyte-colony-stimulating factor (G-CSF) both belong to this group. Trials are still at the commencing stage. In vitro investigations, animal experiments and human experiments have rendered promising results. After intensive chemotherapy followed by treatment with GM-CSF and G-CSF, it has been demonstrated that the period of neutropenia following bone-marrow suppression is abbreviated. Accelerated bone-marrow regeneration and reduced tendencies to infection have been demonstrated compared with so-called control patients. In patients with the myelodysplastic syndrome, marked increase in the neutrophile granulocytes in the peripheral blood has been observed during treatment with growth factors. Long-term treatment with G-CSF has proved particularly effective in chronic idiopathic and cyclic neutropenia. On the other hand, the growth factors can scarcely improve the prognosis in severe aplastic anaemia. Animal experiments and a single human trial suggest that G-CSF and GM-CSF treatment are indicated in bone-marrow insufficiency secondary to radioactive irradiation accidents. GM-CSF has normalized the leucocyte counts in AIDS patients with leucopenia without any marked alteration in the tendency to infection. Future studies will show whether simultaneous treatment with growth factors and anti-viral and anti-leukaemic treatments, respectively, can improve the therapeutic results.


Innate and Peripheral Immune Alterations after Traumatic Brain Injury Are Regulated in a Gut Microbiota-Dependent Manner in Mice.
Traumatic brain injury (TBI) patients are at high risk for disruption of the gut microbiome. Previously, we have demonstrated that broad-spectrum antibiotic exposure after TBI drastically alters the gut microbiota and modulates neuroinflammation, neurogenesis, and long-term fear memory. However, these data did not determine if the impact of antibiotic exposure on the brain's response to injury was mediated directly by antibiotics or indirectly via modulation of the gut microbiota. We designed two different approaches to address this knowledge gap. One was utilizing fecal microbiota transplantation (FMT) from control and antibiotic-treated mice (treated with vancomycin, neomycin, ampicillin, and metronidazole [VNAM]) into germ-free (GF) mice prior to injury, and the other was exposing specific pathogen-free (SPF) mice to a 2-week period of antibiotics prior to injury but discontinuing antibiotics 72 h prior to injury. GF mice receiving FMT from VNAM-treated mice (GF-VNAM) demonstrated reduced gut bacterial alpha diversity and richness compared with GF mice receiving control FMT. At 7 days post-injury, GF-VNAM had increased microglial activation, reduced infiltration of T cells, and decreased neurogenesis. Similarly, SPF mice exposed to antibiotics prior to but not after injury demonstrated similar alterations in neuroinflammation and neurogenesis compared with control mice. These data support our hypothesis implicating the gut microbiota as an important modulator of the neuroinflammatory process and neurogenesis after TBI and provide an exciting new approach for neuroprotective therapeutics for TBI.


Qualification of a whole blood intracellular cytokine staining assay to measure mycobacteria-specific CD4 and CD8 T cell immunity by flow cytometry.
Qualified or validated assays are essential in clinical trials. Short-term stimulation of whole blood and intracellular cytokine staining assay is commonly used to measure immunogenicity in tuberculosis vaccine clinical trials. Previously, the short-term stimulation process of whole blood with BCG was optimized. We aimed to qualify the intracellular cytokine staining process and assess the effects of long-term cryopreservation. Our hypotheses were that the assay is robust in the measurement of the mycobacteria-specific T cells, and long-term cryopreservation of fixed cells from stimulated whole blood would not compromise reliable measurement of mycobacteria induced CD4 T cell immunity. Whole blood from healthy adults was collected in sodium heparinized tubes. The blood was left unstimulated or stimulated with mycobacterial antigens or mitogens for 12h. Cells were harvested, fixed and multiple aliquots from each participant cryopreserved. Later, mycobacteria-specific CD4 and CD8 T cells expressing IFN-γ, TNF-α, IL-2 and IL-17 were quantitated by flow cytometry. Assay performance characteristics evaluated included limit of quantification and detection, reproducibility, precision, robustness, specificity and sensitivity. To assess the effects of long-term cryopreservation, fixed cells from the stimulated bloods were analysed one week post-cryopreservation and at 3-month intervals over a 3-year period. The limit of quantification for the different cytokines was variable: 0.04% for frequencies of IFN-γ- and IL-2-expressing T cells and less than 0.01% for TNF-α- and IL-17-expressing T cells. When measurement of the mycobacteria-specific T cells was assessed at levels above the detection limit, the whole blood intracellular cytokine assay showed high precision that was operator-independent. The assay was also robust: variation in staining conditions including temperature (4 °C or 20-23 °C) and time (45, 60 or 90 min) did not markedly affect quantification of specific T cells. Finally, prolonged periods of cryopreservation also did not significantly influence quantification of mycobacteria-specific CD4 T cells. The whole blood intracellular cytokine assay is robust and reliable in quantification of the mycobacteria-specific T cells and is not significantly affected by cryopreservation of fixed cells.


Magnetic digital flop of ferroelectric domain with fixed spin chirality in a triangular lattice helimagnet.
The ferroelectric properties in a magnetic field (H) of varying magnitude and direction have been investigated for the triangular-lattice helimagnet CuFe(1-x)Ga(x)O(2) (x = 0.035). The in-plane H was found to induce the rearrangement of six possible multiferroic domains. Upon every 60 degrees rotation of in-plane H around the c axis, a unique 120 degrees flop of electric polarization occurs as a result of the switch of the helical magnetic q vector. The chirality of the spin helix is always conserved upon the q flop. The possible origin is discussed in light of the stable structure of the multiferroic domain wall.


Retinal and choroidal alterations in patients with anorexia nervosa without vision loss.
Evaluate the alterations of retinal anatomy and function, as well as choroidal thickness changes, in patients with anorexia nervosa (AN). 13 female AN patients (26 eyes) and 20 female controls (40 eyes) were included. Assessment of the retinal and choroidal anatomy was performed by optical-coherence-tomography, while multifocal-electroretinogram was used for measurements of the electrical activity of the macula. Statistical analysis was performed using t-test. Central macular thickness was found to be thinner in AN (140.04 ± 14.45, 150.85 ± 16.03, p = 0.007), likewise the ganglion cell complex and outer retinal layer (ORL) at most areas. ORL superiorly was thicker in AN (169.12 ± 2.55, 163.00 ± 8.70, p < 0.001), while retinal nerve fiber layer was not different between the two groups, except the inferior region (121.08 ± 18.52, 137.60 ± 7.30, p < 0.001). Choroidal thickness was evenly diminished in the anorectic group (p = 0.001-0.027). Multifocal-electroretinogram showed lower P1-retinal response density amplitude of ring 1 in AN (159.04 ± 60.83, 292.43 ± 11.59, p < 0.001), but no significant difference concerning the P1-response density amplitude of ring 2 (79.04 ± 21.89, 82.63 ± 9.10, p = 0.36). In AN patients, even without vision loss, significant changes occur in retinal and choroidal thickness, as well as in the electrical activity of the macula. However, this is a pilot study in a small sample that needs replication.


Hyperspectral imaging enables the differentiation of differentially inflated and perfused pulmonary tissue: a proof-of-concept study in pulmonary lobectomies for intersegmental plane mapping.
The identification of the intersegmental plane is a major interoperative challenges during pulmonary segmentectomies. The objective of this pilot study is to test the feasibility of lung perfusion assessment by Hyperspectral Imaging for identification of the intersegmental plane. A pilot study (clinicaltrials.org: NCT04784884) was conducted in patients with lung cancer. Measuring tissue oxygenation (StO2; upper tissue perfusion), organ hemoglobin index (OHI), near-infrared index (NIR; deeper tissue perfusion) and tissue water index (TWI), the Hyperspectral Imaging measurements were carried out in inflated (Pvent) and deflated pulmonary lobes (PnV) as well as in deflated pulmonary lobes with divided circulation (PnVC) before dissection of the lobar bronchus. A total of 341 measuring points were evaluated during pulmonary lobectomies. Pulmonary lobes showed a reduced StO2 (Pvent: 84.56% ± 3.92 vs. PnV: 63.62% ± 11.62 vs. PnVC: 39.20% ± 23.57; p<0.05) and NIR-perfusion (Pvent: 50.55 ± 5.62 vs. PnV: 47.55 ± 3.38 vs. PnVC: 27.60 ± 9.33; p<0.05). There were no differences of OHI and TWI between the three groups. This pilot study demonstrates that HSI enables differentiation between different ventilated and perfused pulmonary tissue as a precondition for HSI segment mapping.


Regional epidemiologic assessment of prevalent periodontitis using an electronic health record system.
An oral health surveillance platform that queries a clinical/administrative data warehouse was applied to estimate regional prevalence of periodontitis. Cross-sectional analysis of electronic health record data collected between January 1, 2006, and December 31, 2010, was undertaken in a population sample residing in Ladysmith, Wisconsin. Eligibility criteria included: 1) residence in defined zip codes, 2) age 25-64 years, and 3) ≥1 Marshfield dental clinic comprehensive examination. Prevalence was established using 2 independent methods: 1) via an algorithm that considered clinical attachment loss and probe depth and 2) via standardized Current Dental Terminology (CDT) codes related to periodontal treatment. Prevalence estimates were age-standardized to 2000 US Census estimates. Inclusion criteria were met by 2,056 persons. On the basis of the American Academy of Periodontology/Centers for Disease Control and Prevention method, the age-standardized prevalence of moderate or severe periodontitis (combined) was 407 per 1,000 males and 308 per 1,000 females (348/1,000 males and 269/1,000 females using the CDT code method). Increased prevalence and severity of periodontitis was noted with increasing age. Local prevalence of periodontitis was consistent with national estimates. The need to address potential sample selection bias in future electronic health record-based periodontitis research was identified by this approach. Methods outlined herein may be applied to refine oral health surveillance systems, inform dental epidemiologic methods, and evaluate interventional outcomes.


Genetic variation in the proteins of human nail.
In a survey of the proteins from human nail, genetic variation has been observed in both the low-sulfur and high-sulfur protein fractions. The low-sulfur proteins were examined by polyacrylamide gel electrophoresis at pH 7 in the presence of sodium dodecyl sulfate, whilst the high-sulfur proteins were characterized by gel electrophoresis at pH 2.6 in 3 M urea. Of 106 samples of apparently normal nail, 69% showed a characteristic pattern of 3 major low-sulfur and 5 major high-sulfur proteins. The remaining samples showed an additional high-sulfur band, and about half of these samples also manifested an additional major low-sulfur band. The proportion of nail samples containing the variant proteins was much higher in some families. Two-dimensional eletrophoresis of the low-sulfur proteins showed five major low-sulfur components in the normal nail with an additional component in the variant sample. Peptide maps of the proteins showed that the additional protein was sufficiently different in amino acid sequence to the other proteins to indicate that a mutation in a structural gene had not occurred. In the high-sulfur protein fractions, over 30 proteins were observed in the two-dimensional electrophoretic patterns and there were at least 3 additional components in the variant fraction.


Molecular analysis of methanogen populations and their interactions within anaerobic sludge digesters.
Knowledge of archaeal population structure, function and interactions is of great interest for a deeper understanding of the anaerobic digestion step in wastewater treatment process, that represents a bottle neck in the optimization of digesters performance. Although culture-independent techniques have enabled the exploration of archaeal population in such systems, their population dynamics and interactions still require further investigation. In the present study, 2646 almost full archaeal 16S rRNA gene sequences retrieved from 22 anaerobic digesters located worldwide were analyzed and classified into 83 Operational Taxonomic Units (OTUs) for Euryarchaeotes and 2 OTUs for Crenarchaeotes. Among the Euryarchaeotes, Methanosarcinales represent the predominant archaeal population (47.5% of total sequences), followed by the ARC I (WSA2) lineage (25.3%), Methanomicrobiales (19.9%) and Methanobacteriales (1.9%). Theses lineages are predominant in nine, five, two and one digesters respectively. However, the remaining 5 digesters show no predominance of any methanogenic group. According to the predominance of theses lineages, 5 digester profiles were distinguished. This study revealed a clear interaction between the 4 methanogenic lineages. A core of 12 OTUs represented by five, four, two and one OTU for Methanosarcinales, Methanomicrobiales, ARC I and Methanobacteriales respectively were quantitatively abundant in at least 50% of the analyzed digesters. 16S rRNA targeted hybridization oligonucleotide probes targeting the predominant OTUs are being developed to follow their population dynamics under various parameters.


[10 days of triple treatment of Helicobacter pylori infection and peptic ulcer. Status after treatment of 4 weeks and of one year].
We treated 143 Helicobacter pylori positive ulcer patients (duodenal ulcer 81, gastric ulcer 24, pyloric ulcer nine, previous duodenal ulcer 27, others two) with a ten days regimen of bismuth subnitrate, oxytetracycline and metronidazole. Four weeks after cessation of treatment the 14C-urea breath test showed that the infection was eradicated in 133 out of 140 patients (95%). The ulcer had healed in 129 (97%) of the H. pylori negative patients. Status at one year after treatment showed that 121 (98.4%) of 123 patients who had become H. pylori negative had remained negative. None of the H. pylori negative patients experienced ulcer relapse. The results show that the treatment is effective and that the rate of recurrence of H. pylori during the first year is very low.


Personality Traits, Coping, Health-related Behaviors, and Cumulative Physiological Health in a National Sample: 10 Year Prospective Effects of Conscientiousness via Perceptions of Activity on Allostatic Load.
Personality traits, coping styles, and health-related behaviors show associations with various aspects of health. However, integrative life-course investigations of pathways by which these factors might affect later cumulative physiological health risk remain sparse. To investigate prospective associations of personality traits via coping styles and health-related behaviors on allostatic load in a national sample. Using data from the Midlife in the United States study (MIDUS; N = 1,054), path analyses were used to test direct and indirect associations (via coping styles, smoking, frequency of alcohol consumption, leisure-time physical activity, and perceptions of activity) of personality traits on a latent measurement model of allostatic load informed by 10 biomarkers associated with cardiovascular, inflammation, glucose, and lipid subsystems. Direct 10 year associations of greater conscientiousness on healthier allostatic load and greater extraversion on less healthy allostatic load were observed. Consistent with hypothesized behavioral pathways, relationships between conscientiousness and extraversion on allostatic load were prospectively mediated by greater perceptions of activity. Physical activity and more frequent alcohol use were associated with healthier allostatic load but did not act as prospective mediators. The results provide further evidence of conscientiousness' standing as a marker of health via cumulative physiological health. Moreover, a greater perception of activity was identified as a pathway through which conscientious individuals experienced healthier physiological profiles over time. Examining a more detailed picture of the psychosocial mechanisms leading to development of health risk, as was found with perceptions of activity, remains an important area for future research.


Film density calibration for computed radiography systems: is the standard three-point procedure accurate?
With computed radiography (CR) systems, laser printer-film processor combinations are normally checked daily by using test film three-point density measurements. In this study, the authors show that the recommended three-point quality assurance procedure can show satisfactory results even though the CR film output behaves in an anomalous manner. This problem can be corrected by using a 16-point calibration procedure. Regular 16-point calibrations should be performed to ensure satisfactory CR system performance.


Diagnosis of physical illness in psychiatric patients using axis III and a standardized medical history.
Psychiatrists' axis III diagnoses of physical illnesses in 357 psychiatric patients were compared with diagnoses by a physician's assistant using a standardized medical history form. The physician's assistant detected nearly three times as many physical illnesses as the psychiatrists. The psychiatrists were significantly more likely to miss diagnoses among older patients and women. Patients who met criteria for depressive disorders appeared to be at greatest risk for undetected illnesses, followed by patients with bipolar disorders and schizophrenia. Patients with a secondary diagnosis of substance abuse had significantly more undetected illness than those who did not abuse substances. The authors suggest that current axis III guidelines are inadequate and that a systematic review of physical health problems should be part of the psychiatric diagnostic assessment.


Glucagon-like peptide (GLP) -2 improved colonizing bacteria and reduced severity of ulcerative colitis by enhancing the diversity and abundance of intestinal mucosa.
The global incidence of ulcerative colitis (UC) continues to increase while it's clinical cure rate remains low. Intestinal mucosal ulcers have segmental distribution and variable severity. Intestinal bacteria are closely related to intestinal immunity and metabolism; however, the relationship between intestinal microbiome profile and the occurrence of UC, as well as the contribution of glucose metabolism, are not well understood. This was investigated in the present study using mucosal biopsies from patients with UC and healthy control subjects. We performed high throughput 16S rRNA gene sequencing to estimate microbiota composition and abundance as well as their association with clinical indices such as lesion severity. The results showed that the diversity and abundance of intestinal microbiota were significantly lower in patients with UC than in healthy subjects; however, these were unrelated to ulcer severity. Serum glucagon-like peptide 2 (GLP-2) level was associated with reduced microbiota diversity and abundance in UC. These results indicate that colonization by specific microbiota is not the main determinant of pathologic status in UC. Additionally, therapeutic strategies that increase GLP-2 levels in intestinal mucosa may be effective in the treatment of UC.


Cultural constructions of health and illness. Recent cultural changes toward a holistic approach.
Nurses have defined their domain as holistic with a focus of promoting health. Cultural conceptualizations of health and illness have been changing toward a more complex understanding enriched by perspectives from many disciplines. Health, a more holistic concept than disease, has invited expanded cognitive constructions that accommodate an understanding of the biology, environment, social position, and the role of the mind, culture, spirituality, race, and sex. These have changed our metaphors of the body and expanded the focus to the health-illness experience. These areas are discussed with implications for nursing.


How to calculate sample size in animal and human studies.
One of the most important statistical analyses when designing animal and human studies is the calculation of the required sample size. In this review, we define central terms in the context of sample size determination, including mean, standard deviation, statistical hypothesis testing, type I/II error, power, direction of effect, effect size, expected attrition, corrected sample size, and allocation ratio. We also provide practical examples of sample size calculations for animal and human studies based on pilot studies, larger studies similar to the proposed study-or if no previous studies are available-estimated magnitudes of the effect size per Cohen and Sawilowsky.


Radiotherapy for early-stage prostate cancer in men under 70 years of age.
To demonstrate that radiotherapy (RT) is a valid alternative to surgery in men ≤70 years old with localized prostate cancer. From 1988 to 2009, 214 patients with T1-2 N0 M0 prostate cancer were treated with RT. The effects of patient- and treatment-related risk factors on toxicity were investigated. Median follow-up was 105 months (range 14.2-180). The 5-, 10-, and 15-year biochemical relapse-free survival for all 214 patients was 80%, 61.9%, and 57.5%, respectively. In bivariate analysis, age (≤65 vs 65-70 years) was not a significant factor for biochemical relapse, while radiation dose was (p = 0.05) in multivariate analysis. Cancer-specific survival rates at 5, 10, and 15 years were 98.4%, 93.2%, and 69.7%, respectively. Median overall survival (OS) was 167 months (95% confidence interval 147.3-186.7). The OS rates at 5, 10, and 15 years were 91.8%, 75.8%, and 42.5%, respectively. Acute genitourinary (GU) and gastrointestinal (GI) toxicities occurred in 105 (49%) and 98 patients (45.8%), respectively, with only 2 cases of grade III GI toxicity. Late GU and GI toxicities occurred in 17 (7.9%) and 20 (9.3%) patients, respectively, with 1 grade III GI toxicity and 2 grade III GU toxicities. Risk factors for late toxicity were age and RT dose and technique, which were unrelated to acute toxicity. Age ≤70 years does not consistently confer a negative prognosis for localized prostate cancer. Radiotherapy appears to be a viable alternative to surgery, offering excellent long-term cancer control.


Evaluation of anterior segment parameters in keratoconic eyes measured with the Pentacam system.
To evaluate the alteration in anterior chamber parameters with the progression of keratoconus using a rotating Scheimpflug camera. Inonu University, Turgut Ozal Medical Center, Ophthalmology Department, Malatya, Turkey. Two hundred sixteen eyes of 123 patients diagnosed with keratoconus and 224 eyes of 112 normal subjects were evaluated with a Pentacam Scheimpflug camera (Oculus Inc.). Keratoconic eyes were divided into 3 groups according to mean keratometer (K) readings: mild (K = less than 47.0 diopters [D]), moderate (K = 47.0 to 52.0 D), and severe (K = 52.0 D or higher). The following parameters were obtained: thinnest corneal thickness (TCT), anterior chamber depth (ACD), corneal volume (CV), anterior chamber angle (ACA), and anterior chamber volume (ACV). One hundred twenty-nine eyes had mild keratoconus, 59 had moderate keratoconus, and 35 had severe keratoconus. There were no statistically significant differences between the keratoconus group and control group in age, sex, or eye distribution (P>.05). With the progression of the disease, TCT and ACD were statistically different between all groups (P<.05). There were statistically significant differences in ACA and CV measurements between the mild keratoconus and severe keratoconus groups (P<.05). Also, CV readings were significantly different between the control group and all keratoconus groups (P<.05), although there was no significant difference in ACA measurements (P>.05). Despite a progressive increase in the ACV with disease progression, the differences between groups were not statistically significant (P>.05) except between the severe keratoconus group and the control group (P<.05). Anterior segment parameters of the eye showed significant alterations with the progression of keratoconus.


[Validity and reliability of the Korean version of the observable indicators of nursing home care quality evaluation instrument].
The purposes of this study were; 1) to test the validity and reliability of the Korean version of Rantz's Observable Indicators of Nursing Home Care Quality Instrument (ONHQ) and 2) to evaluate the quality of Korean nursing homes. The study employed a three-phase methodological research design. 1) The original instrument of Rantz's ONHQ was translated into Korean and modified by Korean nursing home experts. 2) A pilot study using the modified instrument was done in 20 nursing homes to examine inter-rater reliability. 3) The validity and reliability were tested in 98 nursing homes. Seven factors were extracted through factor analysis: 'communication', 'care delivery', 'grooming', 'odor', 'environment-basics', 'environment-access', and 'environment-homelike'. These factors explained 86.07% of the total variance. Cronbach's alpha coefficient of the 30 items was .97 indicating a high internal consistency of the instrument. Inter-rater reliability according to Kappa was .82. The average score of nursing home quality was 112.07 indicating an average range of quality level. The Korean version of the ONHQ was identified as a tool with a high degree of validity and reliability. This tool can be effectively used to assess the quality of nursing homes by professions as well as family members.


Post-traumatic stress disorder and coping styles of war veterans twenty years after the war from Tuzla Canton.
Aim To analyse symptoms of posttraumatic stress disorder and coping strategies of war veterans in Tuzla Canton twenty years after the war in Bosnia and Herzegovina (BiH). Methods The study analysed a group of 120 war veterans from the Tuzla Canton who had experience of the war in BiH. For assessment of posttraumatic stress disorder Harvard Trauma Questionnaire was used, a version for Bosnia and Herzegovina and for assessment of coping styles Life Style Index was used. Results Concerning number of traumatic experiences of war veterans, it was found that they suffered 12 traumatic experiences. Most often traumatic experience was the participation in fighting and shelling (90.0%), knowledge of injuries in combat or landmine injuries of family members or friends (75.8%), exposure to snipers (74.2%). The most important were the symptoms of numbnessemotional numbness (2.62%), the symptoms of intrusion (2.58%) and the severity of the symptoms of PTSD (2.39%). The most common strategy of dealing with veterans of war was a projection (68.31%) and intellectualisation (56.20%). Conclusion War veterans have experienced polytraumatic experiences in war and show increased expression of symptoms of posttraumatic stress disorder, emphasised psychosocial problems with a common defence mechanism in the form of projections twenty years after the war. War veterans are in need of continuous treatment in order to reduce long-term consequences of war trauma.


Phenotypic Drug Screening for Dysferlinopathy Using Patient-Derived Induced Pluripotent Stem Cells.
Dysferlinopathy is a progressive muscle disorder that includes limb-girdle muscular dystrophy type 2B and Miyoshi myopathy (MM). It is caused by mutations in the dysferlin (DYSF) gene, whose function is to reseal the muscular membrane. Treatment with proteasome inhibitor MG-132 has been shown to increase misfolded dysferlin in fibroblasts, allowing them to recover their membrane resealing function. Here, we developed a screening system based on myocytes from MM patient-derived induced pluripotent stem cells. According to the screening, nocodazole was found to effectively increase the level of dysferlin in cells, which, in turn, enhanced membrane resealing following injury by laser irradiation. Moreover, the increase was due to microtubule disorganization and involved autophagy rather than the proteasome degradation pathway. These findings suggest that increasing the amount of misfolded dysferlin using small molecules could represent an effective future clinical treatment for dysferlinopathy. Stem Cells Translational Medicine 2019;8:1017-1029.


Deep Learning for Voltammetric Sensing in a Living Animal Brain.
Numerous neurochemicals have been implicated in the modulation of brain function, making them appealing analytes for sensors and diagnostics. However, it is a grand challenge to selectively measure multiple neurochemicals simultaneously in vivo because of their great variations in concentrations, dynamic nature, and composition. Herein, we present a deep learning-based voltammetric sensing platform for the highly selective and simultaneous analysis of three neurochemicals in a living animal brain. The system features a carbon fiber electrode capable of capturing the mixed dynamics of a neurotransmitter, neuromodulator, and ions. Then a powerful deep neural network is employed to resolve individual chemical and spatial-temporal information. With this, a single electrochemical measurement reveals an interplaying concentration changes of dopamine, ascorbate, and ions in living rat brain, which is unobtainable with existing analytical methodologies. Our strategy provides a powerful means to expedite research in neuroscience and empower sensing-aided diagnostic applications.


Scapulothoracic muscle activity and recruitment timing in patients with shoulder impingement symptoms and glenohumeral instability.
Various studies have investigated scapulothoracic muscle activity and recruitment patterns in relation to shoulder complaints in different populations, but a consensus review is lacking. To systematically review the state of the art regarding scapulothoracic muscle activity and recruitment timing in subjects with shoulder pain compared to pain free controls. Systematic review. The search for relevant articles was performed in Pubmed and Web of Science, including Web of Knowledge, using key words related to shoulder pain, scapulothoracic muscle activity or recruitment timing. Articles were included till November 2012. Case-control studies concerning the scapulothoracic region and muscle recruitment using electromyography (EMG) were included. Articles regarding rotator cuff muscles or neck-shoulder pathologies or studies handling a treatment outcome, were excluded. The methodological quality of the articles was assessed using appropriate risk of bias criteria for case-control studies. A total of 12 articles were included in the systematic review, containing patients with Shoulder Impingement Syndrome (SIS) or glenohumeral instability. In patients with SIS 3 out of 6 articles showed increased upper trapezius muscle (UT) activity, 3 out of 5 studies showed decreased lower trapezius muscle (LT) activity and 3 out of 5 articles showed decreased serratus anterior muscle (SA) activity. Patients with glenohumeral instability showed contradictory results on scapulothoracic muscle activity patterns. In both SIS and glenohumeral instability patients, no consensus was found on muscle recruitment timing. Patients with SIS and glenohumeral instability display numerous variations in scapulothoracic muscle activity compared to healthy controls. In the SIS-group, the LT and SA muscle activity is decreased. In addition, the UT muscle activity is increased among the SIS patients, whereas no clear change is seen among patients with glenohumeral instability. Although the scapulothoracic muscle activity changed, no consensus could be made regarding muscle recruitment timing.


Analyzing the neuropsychological characteristics and changes in serum markers of patients with chronic cerebral circulation insufficiency.
To investigate the neuropsychological characteristics and changes in CRP, S100B, MBP, HSP-7, and NSE in serum. Sixty-six (66) patients treated in our hospital as CCCI group were chosen for our study, and 90 patients with depression were selected as the depression group. The patients in both groups were examined with CT perfusion, depression, anxiety and cognition evaluation. Their serum CRP, S100B, MBP, HSP-70 and NSE levels were detected. Neuropsychological and serum markers characteristics were compared. The CBF and CBV in bilateral basal ganglia, frontal lobes, greater oval center, brain stem, and left and right regions of occipital lobes of the patients in CCCI group were significantly lower than in the depression group. The HAMD and HAMA scores of CCCI group patients were significantly lower than in the depression group; CCCI group performed better regarding attention, memory, abstract terms and delayed recall. CCCI also had significantly higher total scores than the depression group. Serum CRP, S100B, MBP, HSP-70 and NSE levels in CCCI group were significantly higher than in the depression group. The differences reach statistical significance (p<0.05). CCCI patients who are accompanied by minor depressive disorder have different degrees of cognitive impairment and experience a significant rise in serum CRP, S100B, MBP, HSP-70 and NSE.


Inter-database validation of a deep learning approach for automatic sleep scoring.
Development of inter-database generalizable sleep staging algorithms represents a challenge due to increased data variability across different datasets. Sharing data between different centers is also a problem due to potential restrictions due to patient privacy protection. In this work, we describe a new deep learning approach for automatic sleep staging, and address its generalization capabilities on a wide range of public sleep staging databases. We also examine the suitability of a novel approach that uses an ensemble of individual local models and evaluate its impact on the resulting inter-database generalization performance. A general deep learning network architecture for automatic sleep staging is presented. Different preprocessing and architectural variant options are tested. The resulting prediction capabilities are evaluated and compared on a heterogeneous collection of six public sleep staging datasets. Validation is carried out in the context of independent local and external dataset generalization scenarios. Best results were achieved using the CNN_LSTM_5 neural network variant. Average prediction capabilities on independent local testing sets achieved 0.80 kappa score. When individual local models predict data from external datasets, average kappa score decreases to 0.54. Using the proposed ensemble-based approach, average kappa performance on the external dataset prediction scenario increases to 0.62. To our knowledge this is the largest study by the number of datasets so far on validating the generalization capabilities of an automatic sleep staging algorithm using external databases. Validation results show good general performance of our method, as compared with the expected levels of human agreement, as well as to state-of-the-art automatic sleep staging methods. The proposed ensemble-based approach enables flexible and scalable design, allowing dynamic integration of local models into the final ensemble, preserving data locality, and increasing generalization capabilities of the resulting system at the same time.


Pharmacokinetics of human recombinant interferon-beta in monkeys and rabbits.
The pharmacokinetics of human recombinant interferon-beta ( ReIFN -beta) was compared with that of natural human interferon-beta (IFN-beta) in monkeys and rabbits after intravenous or intramuscular injection. After intravenous injection of 10(6) units/kg of ReIFN -beta or IFN-beta into monkeys or rabbits, serum levels of both interferons declined biexponentially. No significant differences between ReIFN -beta and IFN-beta were detected in most pharmacokinetic parameters including T1/2-beta, though T1/2-alpha of ReIFN -beta was significantly shorter than that of IFN-beta. These results seemed to be in conflict with the observed difference of stability of the interferons in vitro since ReIFN -beta was less stable than IFN-beta in monkey and rabbit serum. When ReIFN -beta (10(7) units/kg) was injected intramuscularly into monkeys or rabbits, it remained detectable in the serum for 24 hr; an absorption phase and an elimination phase were seen. However, AUC (the area under the serum concentration curve) after the intramuscular injection of ReIFN -beta was about one-half and one-third of that after the intravenous injection in monkeys and rabbits, respectively. ReIFN -beta and IFN-beta (10(6) units/kg) were both detectable in the serum after intramuscular injection into rabbits, but the level of ReIFN -beta was lower than that of IFN-beta. These results indicate that the lack of carbohydrate in ReIFN -beta did not essentially affect the in vivo pharmacokinetics in monkeys and rabbits after intravenous injection, but this was not the case after intramuscular injection.


Drift tube soft-landing for the production and characterization of materials: applied to Cu clusters.
We have recently developed a soft-landing (SL) instrument that is capable of depositing ions onto substrates for preparative and developmental research of new materials using a laser ablation source. This instrument was designed with a custom drift tube and a split-ring ion optic for the isolation of selected ions. The drift tube allows for the separation and thermalization of ions formed after laser ablation through collisions with an inert bath gas. These collisions allow the ions to be landed at energies below 1 eV onto substrates. The split-ring ion optic is capable of directing ions toward the detector or a landing substrate for selected components. Experiments will be shown ablating Cu using an Nd:YAG (1064 and 532 nm) for cluster formation and landing onto a muscovite (mica) surface. The laser ablation of Cu in 8 Torr of He gas gives a spectrum that contains multiple peaks corresponding to Cu(n), Cu(n)O(m) clusters, and their corresponding isomers. Atomic force microscopy and drift tube measurements were performed to characterize the performance characteristics of the instrument.