Classification of subtypes and identification of dysregulated genes in sepsis.
Sepsis is a clinical syndrome with high mortality. Subtype identification in sepsis is meaningful for improving the diagnosis and treatment of patients. The purpose of this research was to identify subtypes of sepsis using RNA-seq datasets and further explore key genes that were deregulated during the development of sepsis. The datasets GSE95233 and GSE13904 were obtained from the Gene Expression Omnibus database. Differential analysis of the gene expression matrix was performed between sepsis patients and healthy controls. Intersection analysis of differentially expressed genes was applied to identify common differentially expressed genes for enrichment analysis and gene set variation analysis. Obvious differential pathways between sepsis patients and healthy controls were identified, as were developmental stages during sepsis. Then, key dysregulated genes were revealed by short time-series analysis and the least absolute shrinkage and selection operator model. In addition, the MCPcounter package was used to assess infiltrating immunocytes. Finally, the dysregulated genes identified were verified using 69 clinical samples. A total of 898 common differentially expressed genes were obtained, which were chiefly related to increased metabolic responses and decreased immune responses. The two differential pathways (angiogenesis and myc targets v2) were screened on the basis of gene set variation analysis scores. Four subgroups were identified according to median expression of angiogenesis and myc target v2 genes: normal, myc target v2, mixed-quiescent, and angiogenesis. The genes CHPT1, CPEB4, DNAJC3, MAFG, NARF, SNX3, S100A9, S100A12, and METTL9 were recognized as being progressively dysregulated in sepsis. Furthermore, most types of immune cells showed low infiltration in sepsis patients and had a significant correlation with the key genes. Importantly, all nine key genes were highly expressed in sepsis patients. This study revealed novel insight into sepsis subtypes and identified nine dysregulated genes associated with immune status in the development of sepsis. This study provides potential molecular targets for the diagnosis and treatment of sepsis.


Amitriptyline-induced ventricular tachycardia: a case report.
In Bangladesh, each emergency physician faces amitriptyline overdose nearly a day. An acute cardiovascular complication, one of the worst complications is mainly responsible for the mortality in tricyclic overdose. Recently, we managed ventricular tachycardia in a young female presented with an impaired consciousness 10 h after intentionally ingesting 2500 mg amitriptyline. Here, we report it, discuss how the electrocardiography is vital to acknowledge and predict it and its' complications and also the recent update of the management of it. A young married Bangladeshi-Bengali girl, 25-year-old, having a history of disharmony with her husband, came with an impaired consciousness after intentionally ingesting 2500 mg amitriptyline about 10 h before arrival. There was blood pressure 140/80 mmHg, heart rate 140 beats-per-min, temperature 103 °F, Glasgow coma scale 10/15, wide complex tachycardia with QRS duration of 178 ms in electrocardiography, blood pH 7.36. Initially, treated with 100 ml 8.4% sodium bicarbonate. After that, QRS duration came to 100 ms in electrocardiography within 10 min of infusion. To maintain the pH 7.50-7.55 over the next 24 h, the infusion of 8.4% sodium bicarbonate consisting of 125 ml dissolved in 375 ml normal saline was started and titrated according to the arterial blood gas analysis. Hence, a total dose of 600 mmol sodium bicarbonate was given over next 24 h. In addition to this, gave a 500 ml intravenous lipid emulsion over 2 h after 24 h of admission as she did not regain her consciousness completely. Afterward, she became conscious, though, in electrocardiography, ST/T wave abnormality persisted. So that, we tapered sodium bicarbonate infusion slowly and stopped it later. At the time of discharge, she was by heart rate 124/min, QRS duration 90 ms in electrocardiogram along with other normal vital signs. Diagnosis of amitriptyline-induced ventricular tachycardia is difficult when there is no history of an overdose obtained. Nevertheless, it should be performed in the clinical background and classic electrocardiographic changes and wise utilization of sodium bicarbonate, intravenous lipid emulsion, and anti-arrhythmic drugs may save a life.


Cytoskeletal reorganization and TPA differently modify AP-1 to induce the urokinase-type plasminogen activator gene in LLC-PK1 cells.
Urokinase-type plasminogen activator (uPA) is an extracellular protease and expressed in various cells that exhibit dynamic changes in cell morphology, suggesting a link between cytoskeletal reorganization (CSR) and uPA expression. CSR can be induced by pharmacological agents, such as by colchicine for microtubule cytoskeleton and by cytochalasin for microfilament cytoskeleton. Using these agents, we previously showed that CSR induced the uPA gene in LLC-PK1 cells independently of the protein kinase C and cAMP-dependent protein kinase. Here we show that the induction of the uPA gene by CSR is mediated by the activation of c-Jun which interacts with an AP-1-like site located 2 kb upstream of the uPA gene. 12-O-tetradecanoylphorbol 13-acetate (TPA) induces the uPA gene through the same elements, but additionally utilizes an adjacent PEA3 element and induces c-fos. Furthermore, CSR induces a greater accumulation and a more pronounced phosphorylation of c-Jun than TPA induction. AP-1 is a positive regulator of growth and oncogenesis, and CSR is an integral part of these processes. Our results provide a view how CSR and AP-1 could be coupled in these processes. We also show that TPA and CSR act synergistically, suggesting a model where an initial activation signal could be amplified by CSR.


Quantum phase slips in superconducting wires with weak inhomogeneities.
Quantum phase slips are traditionally considered in diffusive superconducting wires which are assumed homogeneous. We present a definite estimate for the amplitude of phase slips that occur at a weak inhomogeneity in the wire where local resistivity is slightly increased. We model such a weak link as a general coherent conductor and show that the amplitude is dominated by the topological part of the action. We argue that such weak links occur naturally in apparently homogeneous wires and adjust the estimate to that case. The fabrication of an artificial weak link would localize phase slips and facilitate a better control of the phase-slip amplitude.


Two rapid and sensitive methods based on TaqMan qPCR and droplet digital PCR assay for quantitative detection of Bacillus subtilis in rhizosphere.
Bacillus subtilis, a typical plant growth-promoting rhizobacteria, can benefit plant through promoting growth and reducing disease. The colonization intensity of B. subtilis in rhizosphere is a key factor for improving their effectiveness of field application. In this study, we developed a rapid and sensitive method for detecting B. subtilis in rhizosphere via TaqMan qPCR and droplet digital PCR (ddPCR) methods. The primers/probe set targeting gyrB gene could successfully distinguish B. subtilis from its close-related species. Both the TaqMan qPCR and ddPCR methods exhibited a good linear relationship in the sensitivity assay, suggesting the developed method was specific, effective and reliable. Finally, the two methods were used to detect the colonization dynamic of B. subtilis within Arabidopsis rhizosphere. Both of them showed a consistent trend compared with the traditional cultivation-based and microscopy-based methods. The TaqMan qPCR and droplet digital PCR (ddPCR) methods we developed could be used to rapidly detect B. subtilis in rhizosphere. The TaqMan qPCR and ddPCR methods developed in this study can be applied to rapid quantitative detection of B. subtilis populations, and will be helpful to evaluate their effectiveness of field application.


One does not fit all: Health audience segmentation and prediction of health behaviors in cancer prevention.
The purpose of this study is to propose a health belief model-based approach to segmenting health audiences in order to improve targeting of cancer prevention efforts. This study also examines how the identified health segments differ in cancer prevention behaviors. Using data from the 2013 Health Information National Trends Survey, a cluster analysis resulted in three distinct health audience groups: (a) health aware, (b) health at risk, and (c) health in confidence. MANOVA tests indicate that these segments differ significantly regarding healthy diet and exercise. The findings inform cancer prevention campaigns targeting priority audiences.


Airborne trace elements near a petrochemical industrial complex in Thailand assessed by the lichen Parmotrema tinctorum (Despr. ex Nyl.) Hale.
Several trace elements discharged by the petrochemical industry are toxic to humans and the ecosystem. In this study, we assessed airborne trace elements in the vicinity of the Map Ta Phut petrochemical industrial complex in Thailand by transplanting the lichen Parmotrema tinctorum to eight industrial, two rural, and one clean air sites between October 2013 and June 2014. After 242 days, the concentrations of As, Cd, Co, Cr, Cu, Hg, Mo, Ni, Pb, Sb, Ti, V, and Zn in lichens at most industrial sites were higher than those at the rural and the control sites; in particular, As, Cu, Mo, Sb, V, and Zn were significantly higher than at the control site (p < 0.05). Contamination factors (CFs) indicated that Cd, Cu, Mo, and Sb, which have severe health impacts, heavily contaminated at most industrial sites. Principal component analysis (PCA) showed that most elements were associated with industry, with lesser contributions from traffic and agriculture. Based on the pollution load indexes (PLIs), two industrial sites were highly polluted, five were moderately polluted, and one had a low pollution level, whereas the pollution load at the rural sites was comparable to background levels. This study reinforces the utility of lichens as cost-effective biomonitors of airborne elements, suitable for use in developing countries, where adequate numbers of air monitoring instruments are unavailable due to financial, technical, and policy constraints.


Enhanced performance of high Al-content AlGaN MSM photodetectors by electrode modification using hexadecanethiol.
A metal electrode modification process for AlGaN-based metal-semiconductor-metal (MSM) photodetectors have been introduced to enhance the response of solar-blind ultraviolet (UV) light detection. The hexadecanethiol organic molecules are chemically adsorbed on the electrodes of high-Al-content Al0.6Ga0.4N MSM solar-blind UV photodetectors, which can reduce the work function of the metal electrode and change the height of the Schottky barrier. This modification process significantly increases the photocurrent and responsivity of the device compared with the referential photodetector without modification. Additionally, the adverse effects caused by the surface state and polarization of the AlGaN materials are effectively reduced, which can be beneficial for improving the electrical performances of III-nitride-based UV photodetectors.


Erythrocyte deformation in high-throughput optical stretchers.
Optical stretchers can be used to quantify elastic and homeostatic properties of cells. Because they can apply forces to cells without requiring direct contact, they may noninvasively measure mechanical properties related to cell and membrane health. Present-day optical stretchers are, however, limited to measurements on individual stationary cells, limiting throughput. To overcome this limitation and allow study of variations in cell populations, we recently developed and tested a microfluidic chamber that measures optical stretching parameters for erythrocytes under dynamic flowing conditions. The method uses a single linear diode laser bar and permitted measurements at low flow rates and higher throughput. Here, we numerically investigate the feasibility of further increasing the measurement rates of the optical stretcher in parameter domains where hydrodynamic and optical forces are of comparable magnitude. To do this we couple a recently implemented dynamic optical ray-tracing technique with a fluid-structure interaction solver to simulate the deformation of osmotically swollen erythrocytes in fluid flow of variable rate. Our results demonstrate that a detectable steady-state stretch is induced at nominal optical powers and flow rates. In addition, we find that flow rates can be increased significantly with no major effect on net cell stretch showing the feasibility of application of this technique at greatly increased throughputs.


Antiarrhythmic effect of hypertonic sodium bicarbonate and sodium chloride in ouabain-induced arrhythmias.
Ouabain-induced tachyarrhythmias in cats can be abolished by bolus injection of NaHCO3 solution. This effect is consistent, rapid and of short duration. The antiarrhythmic effect appears to be due to high sodium concentration of this solution, as hypertonic NaCl solution with equal concentration of sodium exerts the same effect. Alkalinity and hyper-osmolality cannot account for the antiarrhythmic effect of NaHCO3 as respiratory alkalosis and mannitol solution with equal osmolality did not relieve the arrhythmias. Overdrive suppression is not responsible for this effect as the heart rate slows down during normalization of the cardiac rhythm. It is postulated that high Na+ concentration abolished ouabain-induced arrhythmias by competition with calcium ion in the myocardial cell sarcotubular system thus replacing it. This hypothesis is supported by the known effect of other agents such as sodium-EDTA and magnesium chloride, in digitalis-induced arrhythmias, which exert their antiarrhythmic properties by removal or displacement of calcium in the myocardial cells.


Primary intraocular lymphoma of T-cell type: report of a case and review of the literature.
Primary intraocular lymphoma (PIOL) is an uncommon non-Hodgkin lymphoma and is usually of B-cell type. Intraocular T-cell or T/NK-cell lymphomas are extremely rare and mostly represent a secondary manifestation of either a cutaneous or a systemic lymphoma. The aim of the current paper is to report the clinical, histopathological and molecular biological findings of a PIOL of T-cell type. Conventional cytological and immunocytological examination of vitrectomy specimens. Conventional histology, immunohistochemistry and polymerase chain reaction (PCR) for the detection of immunoglobulin heavy chain (IgH) and T-cell-receptor gamma (TCR-gamma) gene rearrangement, GeneScan analysis, and DNA sequencing were performed on the chorioretinal biopsy. Cytology of the right vitreous aspirate revealed a moderate cellular infiltrate consisting of medium-sized T-cells with pleomorphic nuclei. Similar atypical lymphocytes were seen in the partially necrotic chorioretinal biopsy. These lymphocytes expressed CD3, CD4, betaF1 and CD30, with a growth fraction of 90%. TCR-gamma-PCR, GeneScan analysis and DNA sequencing demonstrated a monoclonal amplification product within the expected range. In contrast, IgH-PCR revealed oligoclonal amplificates. The patient was treated with low-dose radiotherapy (total 45 Gy), and was in complete remission at final follow-up. A rare PIOL of T-cell type was diagnosed on the basis of vitreous aspiration and chorioretinal biopsy. In addition to conventional cytology and immunocytology, the utilisation of gene rearrangement studies on vitreous or chorioretinal biopsies increases the chances of diagnosing or excluding a PIOL of either B-cell or T-cell type. Despite its rarity, ophthalmic pathologists should always consider the diagnosis of T-PIOL when reviewing vitreous samples.


Type-2 innate lymphoid cells in asthma and allergy.
Type-2 innate lymphoid cells (ILC2) belong to an expanding family of innate lymphocytes that provide a potent source of immune effector cytokines at the initiation of immune responses. ILC2 arise, under the control of the transcription factors RORα and GATA3, from lymphoid progenitors in the bone marrow, to secrete type-2 cytokines including IL-5 and IL-13. Using experimental models, ILC2 have been implicated in allergic diseases, such as asthma and atopic dermatitis, but also in metabolic homeostasis. Furthermore, recent reports have indicated that ILC2 not only play roles at the initiation of type-2 immunity but can also contribute to chronic pathology, such as fibrosis, and can impact on the priming of the adaptive T-cell response. The identification of ILC2 in patients with allergic dermatitis and allergic rhinitis indicates that these cells may represent new therapeutic targets.


Hyperglycaemia confers resistance to chemotherapy on breast cancer cells: the role of fatty acid synthase.
The prognosis for women with breast cancer is adversely affected by the comorbidities of obesity and diabetes mellitus (DM), which are conditions associated with elevated levels of circulating fatty acids, hyperglycaemia and hyperinsulinaemia. We investigated the effects of exposure of non-malignant and malignant human breast epithelial cells to elevated levels of fatty acids and glucose on their growth, survival and response to chemotherapeutic agents. We found that palmitate induced cell death in the non-malignant cells but not in the malignant cells, which was abrogated through the inhibition of ceramide production and by oleate but not by IGF1. Fatty acid synthase (FAS) is responsible for the de novo synthesis of fatty acids from sugars, and is over-expressed in many epithelial cancers. Abundance of FAS was higher in malignant cells than in non-malignant cells, and was up-regulated by IGF1 in both cell types. IGF-induced growth of non-malignant cells was unaffected by suppression of FAS expression, whereas that of malignant cells was blocked as was their resistance to palmitate-induced cell death. Palmitate did not affect cell proliferation, whereas oleate promoted the growth of non-malignant cells but had the opposite effect, that is, inhibition of IGF1-induced growth of malignant cells. However, when the phosphatidylinositol 3-kinase pathway was inhibited, oleate enhanced IGF1-induced growth in both cell types. Hyperglycaemia conferred resistance on malignant cells, but not on non-malignant cells, to chemotherapy-induced cell death. This resistance was overcome by inhibiting FAS or ceramide production. Understanding the mechanisms involved in the associations between obesity, DM and breast cancer may lead to more effective treatment regimens and new therapeutic targets.


Aspirin attenuates the initiation but not the progression of atherosclerosis in apolipoprotein E-deficient mice fed a high-fat, high-cholesterol diet.
Aspirin has potent antiinflammatory properties and attenuates atherosclerosis in apolipoprotein-E-deficient mice fed a high-fat, high-cholesterol diet. In an attempt to clarify the contradictory results obtained with normal chow, we studied the effect of aspirin for a prolonged period of time. The mice were fed a commercial chow until the experiment began at 8 weeks of age. Blood samples were then obtained and several mice (n=8) were sacrificed. The diet of the remaining 48 animals was supplemented with 200 g/kg palm fat and 1 g/kg cholesterol. They were then randomly divided into 2 groups, one of which received 0.5 mg/day of aspirin. The aspirin had a time-dependent effect. First, the extent of lesion decreased; then the effect was neutral; and, finally, after longer periods of being fed the atherogenic diet and receiving aspirin, the extent of the lesion increased. The transitory effect of aspirin should be elucidated in the absence of high dietary lipids.


Long Non-coding RNA Linc01224 Regulates Hypopharyngeal Squamous Cell Carcinoma Growth Through Interactions with miR-485-5p and IGF2BP3.
Increasing evidence illustrates that long non-coding RNAs (lncRNAs) play significant oncogenic roles, including hypopharyngeal squamous cell carcinoma (HSCC). The function and mechanism of long non-coding RNAs (lncRNAs) in hypopharyngeal squamous cell carcinoma (HSCC) have not been fully elucidated. Therefore, this study aimed to investigate the role of a specific lncRNA, linc01224, in regulating the miR-485-5p/IGF2BP3 axis in HSCC. We confirmed the lncRNA expression profiles in 5 pairs of HSCC and normal tissues by lncRNA sequencing. Another 28 HSCC tissues were further validated by quantitative real-time PCR (qRT-PCR). qRT-PCR was also used to detect the expression levels of linc01224, miR-485-5p and IGF2BP3 in HSCC cell lines. Next, functional experiments in vitro and in vivo were applied to determine the effects of linc01224 silencing on tumor proliferation, migration, apoptosis and progression in HSCC. Linc01224 expression was significantly higher in HSCC tissues than in adjacent normal tissues. In addition, HSCC patients with low IGF2BP3 expression had good survival. In vitro assays were mechanistically performed to explore whether linc01224 positively regulates IGF2BP3 expression via its competitive inhibition of miR-485-5p. An in vivo animal model also confirmed that linc01224 could promote the occurrence and development of HSCC. Our study first identified that linc01224 plays an oncogenic role in HSCC. It suggests that linc01224 may act as a prognostic biomarker and potential therapeutic target for HSCC.


Stigma and mental health in endometriosis.
This review provides an overview of patient experiences of endometriosis, endometriosis-related types and sources of stigma pertaining to menstruation, chronic pain, and infertility, and their impact on patients' mental health with particular implications for patient care. Because endometriosis is a complex disease with multifactorial etiology, complicated pathophysiology, and a spectrum of clinical features, diagnosis of endometriosis is typically a lengthy process, and many patients experience initial misdiagnosis. A hallmark symptom is severe menstrual pain with other symptoms including chronic pelvic pain, dysmenorrhea, and infertility. Prior research documents that the diagnostic odyssey, complex management, disabling and unpredictable nature of the disease, and painful symptom profile affect multiple life domains of patients, resulting in poor physical, social, and psychological functioning and clinically-significant rates of anxiety and depression for many. More recently, stigma has been recognized as a potent contributor to poor mental health in endometriosis patients, but existing research is limited and largely atheoretical. We identify major sources of stigma related to endometriosis, including menstrual stigma, chronic pain stigma, and infertility stigma, and their likely impact on patients and health care provision. An integrative theoretical approach is described to facilitate research on the prevalence and effects of endometriosis stigma and their explanatory mechanisms, highlighting specific well-validated psychological instruments to assess stigma. Implications for patient care are emphasized. Better understanding of stigma and mental health in people with endometriosis will enhance the standard of care for this patient population.


Antisense Oligonucleotide-Mediated Exon-skipping Therapies: Precision Medicine Spreading from Duchenne Muscular Dystrophy.
In 1995, we were the first to propose antisense oligonucleotide (ASO)-mediated exon-skipping therapy for the treatment of Duchenne muscular dystrophy (DMD), a noncurable, progressive muscle-wasting disease. DMD is caused by deletion mutations in one or more exons of the DMD gene that shift the translational reading frame and create a premature stop codon, thus prohibiting dystrophin production. The therapy aims to correct out-of-frame mRNAs to produce in-frame transcripts by removing an exon during splicing, with the resumption of dystrophin production. As this treatment is recognized as the most promising, many extensive studies have been performed to develop ASOs that induce the skipping of DMD exons. In 2016, an ASO designed to skip exon 51 was first approved by the Food and Drug Administration, which accelerated studies on the use of ASOs to treat other monogenic diseases. The ease of mRNA editing by ASO-mediated exon skipping has resulted in the further application of exon-skipping therapy to nonmonogenic diseases, such as diabetes mellites. Recently, this precision medicine strategy was drastically transformed for the emergent treatment of only one patient with one ASO, which represents a future aspect of ASO-mediated exon-skipping therapy for extremely rare diseases. Herein, the invention of ASO-mediated exon-skipping therapy for DMD and the current applications of ASO-mediated exon-skipping therapies are reviewed, and future perspectives on this therapeutic strategy are discussed. This overview will encourage studies on ASO-mediated exon-skipping therapy and will especially contribute to the development of treatments for noncurable diseases.


PSD-93 Attenuates Amyloid-β-Mediated Cognitive Dysfunction by Promoting the Catabolism of Amyloid-β.
Amyloid-β (Aβ) is a key neuropathological hallmark of Alzheimer's disease (AD). Postsynaptic density protein 93 (PSD-93) is a key scaffolding protein enriched at postsynaptic sites. The aim of the present study was to examine whether PSD-93 overexpression could alleviate Aβ-induced cognitive dysfunction in APPswe/PS1dE9 (APP/PS1) mice by reducing Aβ levels in the brain. The level of PSD-93 was significantly decreased in the hippocampus of 6-month-old APP/PS1 mice compared with that in wild-type mice. Following lentivirus-mediated PSD-93 overexpression, cognitive function, synaptic function, and amyloid burden were investigated. The open field test, Morris water maze test, and fear condition test revealed that PSD-93 overexpression ameliorated spatial memory deficits in APP/PS1 mice. The facilitation of long-term potentiation induction was observed in APP/PS1 mice after PSD-93 overexpression. The expression of somatostatin receptor 4 (SSTR4) and neprilysin was increased, while the amyloid plaque load and Aβ levels were decreased in the brains of APP/PS1 mice. Moreover, PSD-93 interacted with SSTR4 and affected the level of SSTR4 on cell membrane, which was associated with the ubiquitination. Together, these findings suggest that PSD-93 attenuates spatial memory deficits and decreases amyloid levels in APP/PS1 mice, which might be associated with Aβ catabolism, and overexpression of PSD-93 might be a potential therapy for AD.


Living with diabetes: issues for nursing practice.
Discussion in this paper is based on findings drawn from research undertaken within a qualitative framework of analysis, and with the purpose of exploring people's perceptions of living with diabetes, including the nursing care they receive. A major theoretical concern is to demonstrate the importance of placing patients' and clients' experiences at the centre of nursing interest and practice. It is argued that understanding a disease process should of necessity include an understanding of the concerns of those who live with it. Findings related to the following research questions are discussed: first, what are the challenges of living with diabetes? and second, what are people's perceptions of the nursing care they receive?


Inter-laboratory method validation of CD34+ flow-cytometry assay: the experience of Turin Metropolitan Transplant Centre.
The Turin Metropolitan Transplant Centre (CIC 305) includes four flow-cytometry laboratories assessing quality control on hematopoietic stem cells (HSC) with different instruments and operators. Therefore, the CD34+ enumeration assay should be validated on a regular basis. We describe here the validation plan to test the inter-laboratory reproducibility of CD34+ enumeration assay, based on the risk analysis. Stabilized blood samples were analysed using Stem-Kit reagent according to manufacturer's instructions and acquired using the Beckman Coulter Navios at Regina Margherita Children's' Hospital (305-1), Beckman Coulter FC500 at Candiolo Cancer Institute FPO-IRCCS (305-2), BD Biosciences FACSLyric™ at S. Luigi Hospital (305-3), and Beckman Coulter Navios EX at Mauriziano Hospital (305-4). The ISHAGE guidelines were followed for estimating % and absolute number of CD34+ cells in single-platform method. For each sample repeatability limit (r), reproducibility error, uncertainty of reproducibility error and coefficient of variation (CV) were reported. The repeated measurements from each laboratory or instrument have a variability, expressed as reproducibility error, lower than the repeatability limit for that single parameter. The corrected reproducibility error is always lower than the repeatability limit except for the percentage value of the "low" count. The analysis of inter-laboratory variance is within the maximum acceptable variance value, and the CV of all measurements for each parameter is less than 8%, indicating low measurement variability among laboratories. Evaluating the overall data, we can conclude that the four laboratories are perfectly aligned and the results are reproducible.


A dual delivery of substance P and bone morphogenetic protein-2 for mesenchymal stem cell recruitment and bone regeneration.
Implantation of ex vivo expanded and osteogenically differentiated mesenchymal stem cells (MSCs) for bone regeneration has drawbacks for clinical applications, such as poor survival of implanted cells and increased treatment expenses. As a new approach for bone regeneration that can circumvent these limitations, we propose dual delivery of substance P (SP) and bone morphogenetic protein-2 (BMP-2) to facilitate endogenous stem cell recruitment to bone defects by SP and subsequent in situ osteogenic differentiation of those cells by BMP-2. A heparin-conjugated fibrin (HCF) gel enabled dual delivery with fast release of SP and slow release of BMP-2, which would be ideal for prompt recruitment of endogenous stem cells in the first stage and time-consuming osteogenic differentiation of the recruited stem cells in the second stage. The HCF gels with SP and/or BMP-2 were implanted into mouse calvarial defects for 8 weeks. Local delivery of SP to the calvarial defects using HCF gel was more effective in recruiting MSCs to the calvarial defects than intraperitoneal or intravenous administration of SP. Many of the cells recruited by SP underwent osteogenic differentiation through local delivery of BMP-2. The efficacy of in vivo bone regeneration was significantly higher in the SP/BMP-2 dual delivery group. The dual delivery of SP and BMP-2 using the HCF gel therefore has potential as an effective bone regeneration strategy.


Fine mapping of an epitope recognized by an invasion-inhibitory monoclonal antibody on the malaria vaccine candidate apical membrane antigen 1.
Antibodies that inhibit red blood cell invasion by the Plasmodium merozoite block the erythrocytic cycle responsible for clinical malaria. The invasion-inhibitory monoclonal antibody (mAb) 4G2 recognizes a conserved epitope in the ectodomain of the essential Plasmodium falciparum microneme protein and vaccine candidate, apical membrane antigen 1 (PfAMA1). Here we demonstrate that purified Fab fragments of 4G2 inhibit invasion markedly more efficiently than the intact mAb, suggesting that the invasion-inhibitory activity of this mAb is not due solely to steric effects and that the epitope lies within a functionally critical region of the molecule. We have taken advantage of a synthetic gene encoding a modified form of PfAMA1, and existing x-ray crystal structure data, to fully characterize this epitope. We first validate the gene by demonstrating that it fully complements the function of the authentic gene in P. falciparum. We then use it to identify a group of residues within the previously described domain II loop of PfAMA1 that are critical for recognition by mAb 4G2 and demonstrate that the epitope lies exclusively within this loop with no contributions from residues in other domains of the molecule. This is the first complete characterization of a conserved invasion-inhibitory epitope on PfAMA1. Our results will aid in the design of subunit vaccines designed to generate a broadly effective, focused anti-PfAMA1 protective immune response and may help elucidate the function of PfAMA1.


Towards the simplest hydrodynamic lattice-gas model.
It has been known since 1986 that it is possible to construct simple lattice-gas cellular automata whose hydrodynamics are governed by the Navier-Stokes equations in two dimensions. The simplest such model heretofore known has six bits of state per site on a triangular lattice. In this work, we demonstrate that it is possible to construct a model with only five bits of state per site on a Kagome lattice. Moreover, the model has a simple, deterministic set of collision rules and is easily implemented on a computer. In this work, we derive the equilibrium distribution function for this lattice-gas automaton and carry out the Chapman-Enskog analysis to determine the form of the Navier-Stokes equations.


A case of ovarian torsion in a patient carrier of a FSH receptor gene mutation previously affected by spontaneous ovarian hyperstimulation syndrome.
We here report a case of ovarian torsion in a patient with an history of two previous episodes of spontaneous ovarian hyperstimulation syndrome during her two pregnancies. A mutation of follicle-stimulating hormone receptor (FSHr) gene was identified in this patient and in other members of the family. Two years after her successful second pregnancy, the patient showed signs of severe thyroiditis during administration of oral contraceptive, with suppressed TSH and increased thyreoglobulin, in the absence of any abnormalities of the auto-antibodies. In few days, she developed severe pelvic pain and ultrasonographic evidence of increased ovarian volume. She underwent laparoscopy with unilateral adnexectomy for ovarian ischemic necrosis due to adnexal torsion. Our experience suggests that patients' carrier of a mutation of FSHr gene are at risk of ovarian pathologies also when non-pregnant and in the presence of low TSH levels. Further investigations are needed for an appropriate knowledge of typical and atypical manifestations of spontaneous ovarian hyperstimulation syndrome.


Effect of voluntary exercise on maximal oxygen uptake in young female Fischer 344 rats.
The effect of voluntary exercise on maximal oxygen uptake (VO2 max) was studied in young female Fischer 344 rats. After 10 weeks of wheel-running training, the absolute VO2 max and VO2 max relative to body mass increased without a decline in body mass. The running speed eliciting VO2 max, heart and soleus muscle mass, and succinate dehydrogenase (SDH) activity in the soleus muscle also increased. These results suggest that voluntary exercise is an effective means of increasing the aerobic exercise capacity of young female Fischer 344 rats.


Experimental validation of deuterium oxide-mediated antitumoral activity as it relates to apoptosis in murine malignant astrocytoma cells.
Deuterium oxide (D2O), or heavy water, affects a variety of biological activities different from those of water. The authors examined the antitumoral effect of D2O on brain neoplasms and demonstrated D2O-mediated cytotoxicity by using a Rous sarcoma virus-induced murine malignant astrocytoma cell line, RSVM. The mechanism of the observed cytotoxicity may involve D2O-induced apoptosis and cell-cycle modulation. The authors performed an assay with methylthiazol tetrazolium bromide and a trypan blue dye exclusion test to confirm in vitro D2O-mediated cytotoxicity for RSVM cells. At D2O concentrations of 10 to 50%, the cytotoxic effect was dose and time dependent. Flow cytometry analysis revealed programmed cell death (apoptosis) and the accumulation of RSVM cells during the G2/M phase. By applying the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method, fluorescein isothiocyanate-annexin V and propidium iodide double staining, and caspase-family protease activity analysis, the authors demonstrated both DNA fragmentation and enhancement of caspase activity after a 48-hour treatment with D2O, thus indicating that D2O induces apoptosis in RSVM cells. Apoptotic DNA fragmentation was completely abolished by the caspase inhibitor Z-VAD-FMK (benzyloxycarbonil-Val-Ala-Aps-fluoromethylketone). The findings indicate that the caspase activation pathway may be involved in D2Oinduced apoptosis. The authors found that D2O is cytotoxic to malignant astrocytoma cells. The mechanism of D2O-mediated cytotoxicity involved the induction of apoptosis and cell accumulation during the G2/M phase. This D2O-induced apoptosis is modulated through the caspase activation pathway.


Water jump reorientation: from theoretical prediction to experimental observation.
Liquid water is remarkably labile in reorganizing its hydrogen-bond (HB) network through the breaking and forming of HBs. This rapid restructuring, which occurs on the picosecond time scale, is critical not only for many of the pure liquid's special features but also for a range of aqueous media phenomena, including chemical reactions and protein activity. An essential part of the HB network reorganization is water molecule reorientation, which has long been described as Debye rotational diffusion characterized by very small angular displacements. Recent theoretical work, however, has presented a starkly contrasting picture: a sudden, large-amplitude jump mechanism, in which the reorienting water molecule rapidly exchanges HB partners in what amounts to an activated chemical reaction. In this Account, we first briefly review the jump mechanism and then discuss how it is supported by a series of experiments. These studies range from indirect indications to direct characterization of the jumps through pioneering two-dimensional infrared spectroscopy (2D-IR), the power of which accords it a special focus here. The scenarios in which experimental signatures of the jump mechanism are sought increase in complexity throughout the Account, beginning with pure water. Here 2D-IR in combination with theory can give a glimpse of the jumps, but the tell-tale markers are not pronounced. A more fruitful arena is provided by aqueous ionic solutions. The difference between water-water and water-anion HB strengths provides the experimental handle of differing OH stretch frequencies; in favorable cases, the kinetic exchange of a water between these two sites can be monitored. Sole observation of this exchange, however, is insufficient to establish the jump mechanism. Fortunately, 2D-IR with polarized pulses has demonstrated that HB exchange is accompanied by significant angular displacement, with an estimated jump angle similar to theoretical estimates. The Janus-like character of amphiphilic solutes, with their hydrophobic and hydrophilic faces, presents a special challenge for theory and experiment. Here a consensus on the 2D-IR interpretation has not yet been achieved; this lack of accord impedes the understanding of, for example, biochemical solutes and interfaces. We argue that the influence of hydrophobic groups on water jumps is only modest and well accounted for by an excluded volume effect in the HB exchange process. Conversely, hydrophilic groups have an important influence when their HB strength with water differs significantly from that of the water-water HB. The power of 2D-IR is argued to be accompanied by subtleties that can lead to just the opposite and, in our view, erroneous conclusion. We close with a prediction that a hydrophobic surface offers an arena in which the dynamics of "dangling" water OHs, bereft of a HB, could provide a 2D-IR confirmation of water jumps.


Cardiac overexpression of monocyte chemoattractant protein-1 in transgenic mice prevents cardiac dysfunction and remodeling after myocardial infarction.
Myocardial infarction (MI) is accompanied by inflammatory responses that lead to the recruitment of leukocytes and subsequent myocardial damage, healing, and scar formation. Because monocyte chemoattractant protein-1 (MCP-1) (also known as CCL2) regulates monocytic inflammatory responses, we investigated the effect of cardiac MCP-1 overexpression on left ventricular (LV) dysfunction and remodeling in a murine MI model. Transgenic mice expressing the mouse JE-MCP-1 gene under the control of the alpha-cardiac myosin heavy chain promoter (MHC/MCP-1 mice) were used for this purpose. MHC/MCP-1 mice had reduced infarct area and scar formation and improved LV dysfunction after MI. These mice also showed induction of macrophage infiltration and neovascularization; however, few bone marrow-derived endothelial cells were detected in MHC/MCP-1 mice whose bone marrow was replaced with that of Tie2/LacZ transgenic mice. Flow cytometry analysis showed no increase in endothelial progenitor cells (CD34+/Flk-1+ cells) in MHC/MCP-1 mice. Marked myocardial interleukin (IL)-6 secretion, STAT3 activation, and LV hypertrophy were observed after MI in MHC/MCP-1 mice. Furthermore, cardiac myofibroblasts accumulated after MI in MHC/MCP-1 mice. In vitro experiments revealed that a combination of IL-6 with MCP-1 synergistically stimulated and sustained STAT3 activation in cardiomyocytes. MCP-1, IL-6, and hypoxia directly promoted the differentiation of cardiac fibroblasts into myofibroblasts. Our results suggest that cardiac overexpression of MCP-1 induced macrophage infiltration, neovascularization, myocardial IL-6 secretion, and accumulation of cardiac myofibroblasts, thereby resulting in the prevention of LV dysfunction and remodeling after MI. They also provide a new insight into the role of cardiac MCP-1 in the pathophysiology of MI.


Volume of tobacco advertising in African American markets: systematic review and meta-analysis.
African Americans currently bear the greatest burden of morbidity and mortality due to smoking, and exposure to pro-tobacco media messages predicts smoking. This study compared the concentration (proportion of media messages that are for tobacco) and density (pro-tobacco media messages per person) of pro-tobacco media messages between African American and Caucasian markets. We searched Medline (1966 to June 2006), PsychINFO (1974 to June 2006), and CINAHL (1982 to June 2006) for studies from peer-reviewed journals directly comparing the volume of pro-tobacco media messages in African American and Caucasian markets. From each study, we extracted the number of total media messages, the number of tobacco-related messages, and the number of residents living in each market area. We calculated the concentration and density of tobacco advertising in each market. Out of 131 studies identified, 11 met eligibility criteria, including seven comparing billboard/signage in African American and Caucasian markets and four comparing magazine advertising in African American and Caucasian markets. Meta-analysis estimated a pooled odds ratio of 1.7 (95% confidence interval [CI] 1.1, 2.6) for a given billboard being smoking-related in African American vs. Caucasian market areas (i.e., concentration). The pooled rate ratio of the density of smoking-related billboards was 2.6 (95% CI 1.5, 4.7) in African American vs. Caucasian market areas. Magazine data were insufficient for meta-analysis. Available data indicated that African Americans are exposed to a higher volume of pro-tobacco advertising in terms of both concentration and density. These findings have important implications for research, policy measures, and educational interventions involving racial disparities due to tobacco.


[Value of combined detection of ITGA4 and SFRP2 gene methylation in stool DNA in diagnosis and prognostic evaluation of colorectal tumors].
To investigate the value of quantitative detection of ITGA4 and SFRP2 gene methylation in stool DNA for the early diagnosis and prognostic evaluation of colorectal tumors. Real-time PCR was used for quantitative assessment of ITGA4 and SFRP2 gene methylation levels in stool samples of 85 patients with colorectal cancer, 65 patients with colorectal adenoma and 40 healthy subjects. The 3 groups were comparable for age and gender composition. Methylated ITGA4 and SFRP2 promoters were detected in 48.2% and 62.4% of patients with colorectal cancer, respectively, with a combined positivity of 81.2%. ITGA4 and SFRP2 promoter methylation was detected in 23.1% and 43.1% of patients with colorectal adenoma, respectively, with a combined positivity of 69.2%. The positivity rates of ITGA4 and SFRP2 methylation were significantly higher in patients with colorectal cancer than in those with colorectal adenoma (P < 0.001; P= 0.001) and healthy subjects (P < 0.001; P < 0.001). In colorectal cancer group, ITGA4 and SFRP2 promoter methylation levels were correlated with postoperative tumor recurrence in colorectal cancer group, and the relapse-free survival rate was significantly lower in positive patients for ITGA4 and SFRP2 promoter methylation than in the negative patients (P=0.0002; P=0.007). Multivariate analysis with the COX proportional hazard regression model showed that methylation of ITGA4 and SFRP2 gene promoters (P=0.01) and the degree of tumor differentiation (P=0.03) were associated with the recurrence of colorectal cancer, and were independent risk factors for the recurrence of colorectal cancer. Combined detection of ITGA4 and SFRP2 gene methylation levels in stool DNA can improve the early diagnosis rate of colorectal tumor. ITGA4 and SFRP2 promoter methylation and the degree of tumor differentiation are independent risk factors for colorectal cancer recurrence.


A Spectrum of Regression Following Intravitreal Bevacizumab in Retinopathy of Prematurity.
To describe an improved understanding of the regression patterns following off-label intravitreal bevacizumab (IVB) treatment for retinopathy of prematurity (ROP). Retrospective cohort study. All infants treated with IVB for type 1 ROP at a single institution from June 2013 to March 2018 were retrospectively reviewed and the amount of retinal nonperfusion on fluorescein angiogram was calculated. In the 92 eyes of 46 patients analyzed, only 3 eyes (3.3%) reached full vascular maturity. Of the 89 eyes not reaching maturity, 39 eyes (43.8%) had vascular arrest alone (VAA), 34 eyes (38.2%) had vascular arrest with persistent tortuosity (VAT), and 16 eyes (18.0%) had ROP reactivation. Those eyes that reactivated were more likely to be initially classified as having aggressive posterior ROP (P = .004) and of Asian ethnicity (P = .008). There were greater areas of ischemia in eyes with reactivation as compared to VAT and VAA (112.1 mm2 vs 72.5 mm2 vs 56.6 mm2, respectively, P = .007). Younger gestational age at birth was found to be an independent predictor of persistent tortuosity (VAT vs VAA) in a logistic regression model. Incomplete vascularization following IVB is very common and is associated with a younger gestational age at birth, Asian ethnicity, and aggressive posterior ROP. The presence of tortuosity following IVB may be indicative of persistently elevated vascular endothelial growth factor levels and an early indicator of potential reactivation.


Characteristics of human large granular lymphocytes and relationship to natural killer and K cells.
Recent evidence, has demonstrated an association between a subpopulation of peripheral blood mononuclear cells, morphologically identified as large granular lymphocytes (LGL), and natural killer (NK) activity. We have now evaluated more directly the role of LGL in both NK activity and antibody- dependent cellular cytotoxicity (ADCC), by using highly enriched populations of LGL, obtained by centrifugation of peripheral blood mononuclear cells on Percoll discontinuous density gradients. Both spontaneous and interferon- augmented NK and ADCC activities were exclusively associated with the LGL- enriched, low density fractions. The majority of LGL formed conjugates with NK-susceptible and antibody-coated target cells. Approximately 20 percent of small conventional lymphocytes also formed conjugates with the target cells for NK, but this was not associated with cytotoxic activity. Virtually all LGL were found to have receptors for the Fc portion of IgG (FcgammaR). The frequency of LGL among blood leukocytes was 2-6 percent. LGL could be enriched to an average purity of 95 percent by combining discontinuous density gradient centrifugation with subsequent adsorptions of the low density fractions on monolayers of immobilized immune complexes. About 50 percent of LGL were found to be FcgammaR-bearing T cells (T(G)), forming low affinity rosettes with sheep erythrocytes at 4 degrees C. Only 10-20 percent of LGL formed high affinity rosettes with sheep erythrocytes at 29 degrees C. LGL could be enriched to a purity of more than 90 percent by depleting high affinity rosette-forming cells from low density Percoll fractions. LGL were only a subpopulation of T(G) cells, because some lymphocytes with conventional morphology also adhered to the immobilized immune complex monolayers and formed high affinity rosettes with sheep erythrocytes. Separation of these cells from LGL by discontinuous density gradient centrifugation indicated that they are not cytotoxic, suggesting a morphological and functional subdivision of T(G) cells. The verification in this study that virtually all human NK and K cells have a characteristic morphology adds a useful parameter to the monitoring of human lymphocytes, and the ability to purify these cells by simple physical procedures should be invaluable in their further characterization.


Measurement of respiratory tract ammonia in the rabbit and implications to sulfuric acid inhalation studies.
Ammonia (NH3) in the respiratory tract has the potential to neutralize inhaled acid vapors and aerosols. Levels of exhaled (nasal) NH3 were measured in rabbits at different times on the same day, on different days, and in rabbits in a normal fed state, or in a fasted or fed state in which the teeth were brushed and the mouth cleansed. The variability of NH3 levels within any individual rabbit was found to be of the same order as the variability found between different animals. In addition, rabbits which were fasted and had their teeth brushed exhaled significantly less NH3 than did fed animals. Levels in the former group ranged from 4 to 236 micrograms/m3, while those in the latter group ranged from 10 to 758 micrograms/m3. Although brushing the teeth of fed animals compressed the observable range of NH3 levels (22-404 micrograms/m3), this was not a significant reduction compared to fed, unbrushed animals. Thus, fasting likely minimized foodstuff in the mouth; the latter may contribute to NH3 formation through bacterial degradation, which appears to be a significant source of NH3 exhaled through the nose. The NH3 concentrations observed may produce variable degrees of neutralization of inhaled H2SO4 droplets before they deposit in the lungs.


CovR activation of the dipeptide permease promoter (PdppA) in Group A Streptococcus.
CovR, the two-component response regulator of Streptococcus pyogenes (group A streptococcus [GAS]) directly or indirectly represses about 15% of the genome, including genes encoding many virulence factors and itself. Transcriptome analyses also showed that some genes are activated by CovR. We asked whether the regulation by CovR of one of these genes, dppA, the first gene in an operon encoding a dipeptide permease, is direct or indirect. Direct regulation by CovR was suggested by the presence of five CovR consensus binding sequences (CBs) near the putative promoter. In this study, we identified the 5' end of the dppA transcript synthesized in vivo and showed that the start of dppA transcription in vitro is the same. We found that CovR binds specifically to the dppA promoter region (PdppA) in vitro with an affinity similar to that at which it binds to other CovR-regulated promoters. Disruption of any of the five CBs by a substitution of GG for TT inhibited CovR binding to that site in vitro, and binding at two of the CBs appeared cooperative. In vivo, CovR activation of transcription was not affected by individual mutations of any of the four CBs that we could study. This suggests that the binding sites are redundant in vivo. In vitro, CovR did not activate transcription from PdppA in experiments using purified GAS RNA polymerase and either linear or supercoiled DNA template. Therefore, we propose that in vivo, CovR may interfere with the binding of a repressor of PdppA.


Core-Shell Structured o-LiMnO2@Li2CO3 Nanosheet Array Cathode for High-Performance, Wide-Temperature-Tolerance Lithium-Ion Batteries.
To develop a high-capacity, high-rate, cycle-stable cathode material has long been the focus for lithium-ion battery (LIB) research. Recently, layer-structured orthorhombic-LiMnO2 (o-LMO) has attracted extensive interest owing to its large discharge capacities. However, poor cycle performance greatly hinders its practical application, especially at high temperatures. Conventional strategies to address this issue often lead to sacrificed rate performance and mostly work at low temperatures. Herein, we report a novel core-shell structured, o-LiMnO2@Li2CO3 (o-LMO@Li2CO3) nanosheet array cathode, where the Li2CO3 shell improves cycle performance by preventing o-LMO dissolution in the electrolyte (even at an elevated temperature), the o-LMO core provides high capacities and the nanosheet array architecture ensures rate performance (to the best of our knowledge, this o-LMO nanosheet array architecture is reported for the first time). The above features work synergistically to give well-balanced cycle performance (79% capacity retention at 60 °C, 400 cycles), capacity (207 mAh g(-1) at 0.5C) and rate performance (128 mAh g(-1) at 5C) of the o-LMO@ Li2CO3 cathode as well as remarkable full-cell performance (∼67% capacity retention for 400 cycles at ∼2C, 60 °C). Our work demonstrates that the synergistic effect between the o-LMO core, Li2CO3 coating and the nanoarray structure is an effective strategy for developing high-energy/power density, high-stability LIB cathodes.


Healthcare performance and the effects of the binaural beats on human blood pressure and heart rate.
Binaural beats are the differences in two different frequencies (in the range of 30-1000 Hz). Binaural beats are played through headphones and are perceived by the superior olivary nucleus of each hemisphere of the brain. The brain perceives the binaural beat and resonates to its frequency (frequency following response). Once the brain is in tune with the binaural beat it produces brainwaves of that frequency altering the listener's state of mind. In this experiment, the effects of the beta and theta binaural beat on human blood pressure and pulse were studied. Using headphones, three sounds were played for 7 minutes each to 12 participants: the control,- the sound of a babbling brook (the background sound to the two binaural beats), the beta binaural beat (20 Hz), and the theta binaural beat (7 Hz). Blood pressure and pulse were recorded before and after each sound was played. Each participant was given 2 minutes in-between each sound. The results showed that the control and the two binaural beats did not affect the 12 participant's blood pressure or pulse (p > 0.05). One reason for this may be that the sounds were not played long enough for the brain to either perceive and/or resonate to the frequency. Another reason why the sounds did not affect blood pressure and pulse may be due to the participant's age since older brains may not perceive the binaural beats as well as younger brains.


[Microsurgery in urology].
The use of microsurgical techniques offers operative perspectives also in urology. Microsurgery is indicated for reconstructive vascular surgery in the kidneys, in cases of erectile impotence, and in high maldescended testicles, for recanalisation of the vas deferens and in early childhood for plasty surgery of malformations of the upper urinary tract and such of the male genitals. Certain indications are presented under reference to own clinical experiences.


Vitamin D: Current Guidelines and Future Outlook.
Vitamin D is of public health interest because its deficiency is common and is associated with musculoskeletal diseases, as well as extraskeletal diseases, such as cancer, cardiovascular diseases, and infections. Several health authorities have reviewed the existing literature and published nutritional vitamin D guidelines for the general population. There was a wide consensus that serum 25-hydroxyvitamin D [25(OH)D] concentration should be used to assess vitamin D status and intake, and that musculoskeletal, and not extraskeletal, effects of vitamin D should be the basis for nutritional vitamin D guidelines. Recommended target levels for 25(OH)D range from 25 to 50 nmol/l (10 to 20 ng/ml), corresponding to a vitamin D intake of 400 to 800 International Units (10 to 20 μg) per day. It is of concern that significant sections of the general population do not meet these recommended vitamin D levels. This definitely requires action from a public health perspective.


The role of ultrasonic cavitation in refining the microstructure of aluminum based nanocomposites during the solidification process.
Recent studies showed that the microstructure and mechanical properties of aluminum based nanocomposites can be significantly improved when ultrasonic cavitation and solidification processing is used. This is because ultrasonic cavitation processing plays an important role not only in degassing and dispersion of the nanoparticles, but also in breaking up the dendritic grains and refining the as-cast microstructure. In the present study, A356 alloy and Al2O3 nanoparticles are used as the matrix alloy and the reinforcement, respectively. Nanoparticles were added into the molten A356 alloy and dispersed via ultrasonic cavitation processing. Ultrasonic cavitation was applied over various temperature ranges during molten alloy cooling and solidification to investigate the grain structure formation and the nanoparticle dispersion behavior. Optical Microscopy and Scanning Electron Microscopy were used to investigate in detail the differences in the microstructure characteristics and the nanoparticle distribution. Experimental results indicated that the ultrasonic cavitation processing and Al2O3 nanoparticles play an important role for microstructure refinement. In addition, it was shown in this study that the Al2O3 nanoparticles modified the eutectic phase.


Simultaneous quantification of eleven bioactive components of male flowers of Eucommia ulmoides oliver by HPLC and their quality evaluation by chemical fingerprint analysis with hierarchical clustering analysis.
Eucommia ulmoides Oliv (EU), a dioecious perennial angiosperm, is one of the oldest tonics in Chinese traditional medicine. The tea of male flowers of EU has been become popularities and seen as aspirational health care tea in China. There were no enough marks and effective method to control the quality of male flowers of EU. A simple and efficient HPLC method was developed for the simultaneous determination of 11 bioactive compounds (4 iridoids, 1 phenylpropanoid, 6 flavonoids). HPLC chromatographic fingerprint and hierarchical cluster analysis were used to evaluate and classify the samples of male flowers of EU which came from different locations in China. Samples were separated on a Thermal hypersil gold column (250 mm × 4.6 mm, 5 μm) and detected by an ultraviolet detector. The UV wavelength was set at 206, 236, and 206 nm. Mobile phase consisted of methanol (B) and phosphoric acid-water (0.5%) (C) using a gradient elution. Analytes were performed at 25°C with a flow rate of 1.0 mL/min. In quantitative analysis, the eleven components showed good regression (r(2) > 0.9996) within linear ranges, and their recoveries were in the range of 98.65-102.31%. In the chromatographic fingerprint, 16 peaks were selected as the characteristic peaks to assess the similarities of different samples. Hierarchical cluster analysis (HCA) was also applied to differentiate the samples based on the area of all the common peaks. The samples which had higher similarity in HPLC fingerprint were classified as a cluster. This study will provide methodological reference for the quality control and sample classification of male flowers of E. ulmoides.


Toxicity assessment of the extractables from multi-layer coextrusion poly ethylene bags exposed to pH=5 solution containing 4% benzyl alcohol and 0.1 M sodium acetate.
A non-target analysis was developed for the analysis of extractables from multi-layer coextrusion bags exposed to 4% benzyl alcohol solution and 0.1 M sodium acetate at pH = 5 for defined periods (15 day, 45 day and 90 day) according to manufacturer instructions based on the ultra-performance liquid chromatography (UPLC) quadrupole-time of flight mass spectrometry (Q-TOF MS). In order to confirm the extractables, principal component analysis (PCA) was used to indicate the differences among samples of different periods. Then, the extractables were identified based on searching the self-built library or online searching. The total content of extractables of 90 day samples was 589.78 μg/L, and the content was in the range of acceptable levels for pharmaceutical manufacturers. The risk assessment of the extractables were evaluated by Toxtree and T.E.S.T. software to avoid the animals bioexperiment.


A Behavioral analysis of nurses' and pharmacists' role in addressing vaccine hesitancy: scoping review.
The purpose of this review was to identify, characterize, and map the existing knowledge on a) nurses' and pharmacists' perceived barriers and enablers to addressing vaccine hesitancy among patients; and b) strategies or interventions for nurses and pharmacists to address vaccine hesitancy in their practice. Our comprehensive search strategy targeted peer-reviewed and grey literature. Two independent reviewers screened papers and extracted data. We coded narrative descriptions of barriers and enablers and interventions using the Behavior Change Wheel. Sixty-six records were included in our review. Reported barriers (n = 9) and facilitators (n = 6) were identified in the capability, opportunity and motivation components. The majority of the reported interventions were categorized as education (n = 47) and training (n = 26). This current scoping review offers a detailed behavioral analysis of known barriers and enablers for nurses and pharmacists to address vaccine hesitancy and interventions mapped onto these behavioral determinants.


Preferential induction of CYP1A1 and CYP1B1 in CCSP-positive cells.
Both benzo[a]pyrene (BaP) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are potent ligands of aryl hydrocarbon receptors (AhR). Although animal studies indicate that both compounds induce pathological changes in the peripheral lung, the specific cell type involved remains unclear. Clara cells, expressing Clara cell specific protein (CCSP) and abundant in cytochrome P450, are nonciliated bronchiolar epithelial cells in the peripheral lung. Here we explore the hypothesis that CCSP-positive Clara cells are highly responsive to AhR ligands and are the primary cell type involved in BaP- and TCDD-induced toxicities. The responsiveness to AhR ligands was evaluated by measuring the respective mRNA and protein levels of cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1) using real-time RT-PCR and immunocytochemistry assays. Two in vitro models were used: primary cultures of human small airway epithelial (SAE) cells and rat lung slice cultures. In the presence of calcium, human SAE cells differentiated into CCSP-positive cells. BaP- and TCDD-induced mRNA and protein levels of CYP1A1 and CYP1B1 levels were significantly elevated in CCSP-positive cell cultures. Similarly, AhR mRNA and protein levels were increased in CCSP-positive cell cultures, as determined by real-time RT-PCR and Western blot analysis. When rat lung slice cultures were treated with BaP or TCDD for 24 h, CYP1A1 and CYP1B1 proteins were strongly induced in Clara cells. These results indicate that, in the peripheral lung of both rats and humans, CCSP-positive cells (Clara cells) may be more sensitive to AhR ligands than other cell types.


Role of mTORC1-S6K1 signaling pathway in regulation of hematopoietic stem cell and acute myeloid leukemia.
Dysregulation of the mechanistic target of rapamycin complex 1 (mTORC1)-p70 ribosomal protein kinase 1 (S6K1) signaling pathway occurs frequently in acute myeloid leukemia (AML) patients. This pathway also plays a critical role in maintaining normal cellular processes. Given the importance of leukemia stem cells (LSCs) in the development of minimal residual disease, it is critical to use therapeutic interventions that target the LSC population to prevent disease relapse. The mTORC1-S6K1 pathway has been identified as an important regulator of hematopoietic stem cell (HSC) and LSC functions. Both HSC and LSC functions require regulation of key cellular processes including proliferation, metabolism, and autophagy, which are regulated by mTORC1 pathway. Despite the mTORC1-S6K1 pathway being a critical regulator of AML initiation and progression, inhibitors of this pathway alone have yielded mixed results in clinical studies. Recent studies have identified strategies to develop new mTORC1-S6K1 inhibitors such as RapaLink-1, which could circumvent the drug resistance observed in AML cells and in LSCs. Here, we review recent advances made in identifying the role of different components of this pathway in the regulation of HSCs and LSCs and discuss possible therapeutic approaches.


Effect of glucose on insulin biosynthesis and beta cell ultrastructure in cultured fetal rat pancreas.
Pancreatic rudiments from 14-day fetal rats were cultured whole for 8 days in medium containing 5.5 or 16.5 mmol glucose/l (1G or 3G medium). Rudiments grown in 3G medium (3G cells) contained more DNA and insulin than those grown in 1G medium (1G cells) but there was no alteration in the insulin/DNA ratio or the fractional area of the rudiment occupied by insulin-containing cells. Morphometric analysis of ultrastructure revealed that the beta cells grown in 3G medium were smaller and had smaller nuclei than those grown in 1G medium. The size of exocrine cell nuclei in 1G or 3G medium was similar. Insulin granules occupied a greater proportion of the cytoplasmic volume in rudiments grown in 3G medium although the mean absolute volume of insulin granules per cell grown in 1G and 3G media was similar. Hence the residual cytoplasmic volume (cell--nucleus and granules) of 3G cells was less than that of 1G cells. Insulin granules from 3G cells had smaller granule sacs and cores than those from 1G cells. It is concluded that glucose stimulates the growth of rat fetal pancreas in vitro and has important effects on beta cell ultrastructure.


Parathyroid hormone: an early predictor of symptomatic hypocalcemia after total thyroidectomy.
The purpose of this study was to evaluate if the measurement of peri-operative parathyroid hormone (PTH) is able to identify patients with increased risk of developing symptoms of hypocalcemia. Forty patients who underwent total thyroidectomy were studied prospectively. Ionized serum calcium and PTH were measured after induction of anesthesia, one hour (PTH1) and one day after surgery (PTH24). Patients were evaluated for symptoms of hypocalcemia and treated with calcium and vitamin D supplementation as necessary. Symptomatic hypocalcemia developed in 16 patients. Symptomatic patients had significant lower PTH1 and greater drops in PTH levels. The selection of 12.1 ng/L as PTH1 level cutoff level divided patients with and without symptoms with 93.7% sensitivity and 91.6% specificity. The selection of 73.5% as the cutoff value for PTH decrease resulted in 91.6% sensitivity and 87.5% specificity. PTH1 levels and the drop in PTH levels are reliable predictors of developing symptomatic hypocalcemia after total thyroidectomy.


Optimization of black-box models with uncertain climatic inputs-Application to sunflower ideotype design.
Accounting for the interannual climatic variations is a well-known issue for simulation-based studies of environmental systems. It often requires intensive sampling (e.g., averaging the simulation outputs over many climatic series), which hinders many sequential processes, in particular optimization algorithms. We propose here an approach based on a subset selection in a large basis of climatic series, using an ad-hoc similarity function and clustering. A non-parametric reconstruction technique is introduced to estimate accurately the distribution of the output of interest using only the subset sampling. The proposed strategy is non-intrusive and generic (i.e. transposable to most models with climatic data inputs), and can be combined to most "off-the-shelf" optimization solvers. We apply our approach to sunflower ideotype design using the crop model SUNFLO. The underlying optimization problem is formulated as a multi-objective one to account for risk-aversion. Our approach achieves good performances even for limited computational budgets, outperforming significantly standard strategies.


Isomodal column switching high-performance liquid chromatographic technique for the analysis of ciglitazone and its metabolites in human serum.
The application of isomodal column switching high-performance liquid chromatography as an alternative to gradient elution was investigated for the analysis of ciglitazone , a potential oral antidiabetic agent, and its monohydroxyl metabolites in human serum. A high-performance liquid chromatographic apparatus was designed to perform on-line fractionation of the serum extract into non-polar (drug) and polar (metabolite) fractions which were then automatically routed into individually optimized, isocratic, reversed-phase high-performance liquid chromatographic systems for simultaneous analysis. Sample fractionation was performed with a reversed-phase guard column, and solvent routing was accomplished with microprocessor-controlled switching valves. Serum was extracted for analysis by a one-step mode sequencing procedure using disposable bonded-phase columns, and quantitation was accomplished with spiked serum standards. Performance specifications of the method were defined for precision, accuracy, linearity, and sensitivity. The column switching method was found to be both expedient and reliable, and it may have general utility for the routine, quantitative analysis of drug/metabolite mixtures that cannot be assayed by simple isocratic elution methods.


Establishment of loop-mediated isothermal amplification method (LAMP) for the detection of Vibrio nigripulchritudo in shrimp.
LAMP is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. In this study, using the LAMP method, a diagnostic protocol was developed for the detection of Vibrio nigripulchritudo in shrimps. Vibrio nigripulchritudo is associated with distinct shrimp diseases (vibriosis) and is considered one of the threatening pathogens in shrimp industry. After initial cloning and sequencing of the intergenic spacer region (ITS) between 16S and 23S rRNA genes of V. nigripulchritudo, a set of four primers - two inner and two outer - were designed for use in the LAMP reaction. Reaction time and temperature were optimized for 60 min at 63 degrees C, respectively. The detection limit of V. nigripulchritudo by LAMP was 10(2) CFU mL(-1) but PCR could detect up to 10(3) CFU mL(-1). The LAMP method could detect the presence of V. nigripulchritudo from heart, lymphoid organ, and muscle of experimentally infected shrimps with V. nigripulchritudo. This study established a highly sensitive and a rapid diagnostic procedure for detection of V. nigripulchritudo in shrimps. The method developed in this study could be very useful for routine shrimp disease diagnostics.


Accumulation of iodine-131-iodocholesterol in renal cell carcinoma adrenal metastases.
Adrenocortical scintigraphy is a useful technique for differentiating between types of nonhyperfunctioning adrenal masses. Metastatic tumors do not normally accumulate radioiodocholesterol and show discordant uptake on scintigrams. We present two patients who showed accumulation of 131I-6beta-iodomethyl-19-norcholesterol (NP59) in the adrenal metastases from renal cell carcinoma. In one patient with bilateral adrenal metastases, accumulation in the primary tumor as well as adrenal metastases was demonstrated. The adrenal metastases in both patients were resected and were histologically proven to be metastases from clear-cell renal carcinoma. Accumulation of NP59 in metastatic adrenal tumors, although a very rare finding, suggests a pitfall in the differential diagnosis of adrenal cortical tumors.


Schistosomes causing cercarial dermatitis: a mini-review of current trends in systematics and of host specificity and pathogenicity.
The human infection known under the names cercarial dermatitis or swimmers' itch is generally associated with swimming in lakes all over the world, however, a number of outbreaks of cercarial dermatitis developing in salt or brackish waters are also reported. The disease presents as allergic reaction which is able to trap and eliminate the parasites in the skin. However, the infection can be linked to more than skin symptoms under certain circumstances. Recent studies on bird schistosomes have shown that during primary infections of noncompatible hosts (mice) the parasites may migrate through visceral and nervous tissues of mammals. Up to date, cercarial dermatitis has been mostly associated with the cercariae of bird schistosomes of the genus Trichobilharzia. Recent findings of new genera and species indicate, however, broader spectrum of causative agents of the disease with different life cycles, host specificity and pathogenicity.


Initial human experience with the XIENCE side-branch access device.
The everolimus-eluting XIENCE side-branch access (SBA) stent has been the focus of numerous recent publications. Most of the information available on this device comes from the preclinical studies performed in ovine models as well as perfused synthetic heart models. It has now become available in Europe as part of a limited test launch. Delivered via a low-profile, dual-lumen, single-tip catheter, a single inflation device deploys the stent in the main branch and expands a portal opening into the ostium of the side branch to allow for scaffolding and entry into the side branch. This case report describes the first-in-man experience with this novel device.


Antibiotic resistance in Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes in respiratory tract infections in outpatients.
Sensitivity patterns of Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes were studied prospectively in an outpatient population seeking medical advice for respiratory tract infections (RTI) in the Southern parts of Stockholm. In total, 3,214 nasopharyngeal and 1,907 throat swabs were cultured during January-February 1996. 32% of the patients had received antibiotics during the previous year. Reduced penicillin sensitivity in S. pneumoniae was rare (1.3%) and only seen in patients treated with antibiotics during the previous 4 months. Beta-lactamase production in H. influenzae was found in 13.4% of patients who had been treated with antibiotics during the last 4 months and in 7.9% of the others. No resistance (< 1%) to erythromycin was seen in S. pyogenes. In this population-based surveillance, the levels of resistance in common respiratory tract pathogens were thus low and correlated to previous antibiotic treatment. Strict indications for antibiotic treatment in uncomplicated RTI are advocated to maintain a low resistance rate. Penicillin is still the drug of choice in patients without frequent recurrences of RTI in a setting similar to the one studied.


Human factors engineering: the next challenge.
This fictional exchange between a General Manager, a Research and Development Officer and a Regulatory Affairs Manager after a near incident with a recently launched device explains how human factors engineering is the key to design success.


Single-Mode Lasing from "Giant" CdSe/CdS Core-Shell Quantum Dots in Distributed Feedback Structures.
"Giant" semiconductor quantum dots (GQDs) have tremendous potential for applications in laser devices. Here, CdSe/CdS core-shell GQDs (11 monolayers) have been synthesized as lasing gain material. The photoluminescence decay of the GQD ensemble is single-exponential, and the two-photon absorption cross-section is above 105 GM. This article presents a versatile method for fabrication of CdSe/CdS GQD distributed feedback (DFB) lasers by laser interference ablation. A high-quality surface-relief grating structure can be readily created on the GQD thin films, and the relationship between laser beam intensity and surface modulation depth is studied. With appropriate periods, single-mode lasing emission has been detected from these devices under excitation wavelengths of 400 and 800 nm. The laser thresholds are as low as 0.028 and 1.03 mJ cm-2, with the lasing Q-factors of 709 and 586, respectively. Lasing operation is realized from the direct laser interference-ablated QD DFB structures for the first time.


Obesity-related glomerulopathy: Current approaches and future perspectives.
Obesity-related glomerulopathy (ORG) is a silent comorbidity which is increasing in incidence as the obesity epidemic escalates. ORG is associated with serious health consequences including chronic kidney disease, end-stage renal disease (ESRD), and increased mortality. Although the pathogenic mechanisms involved in the development of ORG are not fully understood, glomerular hemodynamic changes, renin-angiotensin-aldosterone system (RAAS) overactivation, insulin-resistance, inflammation and ectopic lipid accumulation seem to play a major role. Despite albuminuria being commonly used for the non-invasive evaluation of ORG, promising biomarkers of early kidney injury that are emerging, as well as new approaches with proteomics and metabolomics, might permit an earlier diagnosis of this disease. In addition, the assessment of ectopic kidney fat by renal imaging could be a useful tool to detect and evaluate the progression of ORG. Weight loss interventions appear to be effective in ORG, although large-scale trials are needed. RAAS blockade has a renoprotective effect in patients with ORG, but even so, a significant proportion of patients with ORG will eventually progress to ESRD despite therapeutic efforts. It is noteworthy that certain antidiabetic agents such as sodium-glucose cotransporter 2 inhibitors (SGLT2i) or glucagon-like peptide-1 receptor agonists (GLP-1 RAs) could be useful in the treatment of ORG through different pleiotropic effects. In this article, we review current approaches and future perspectives in the care and treatment of ORG.


Mapping of a locus (mdoA) that affects the biosynthesis of membrane-derived oligosaccharides in Escherichia coli.
Mutants of Escherichia coli defective in the newly discovered mdoA locus are blocked at an early stage in the biosynthesis of membrane-derived oligosaccharides. The mutation has now been mapped and found to be located near 23 min on the E. coli chromosome between putA and pyrC. The mdoA mutants are defective in the membrane-localized component of the glucosyl transferase system described by Weissborn and Kennedy (A. C. Weissborn and E. P. Kennedy, Fed. Proc. 42:2122, 1983).


Imaging of carbon nanomembranes with helium ion microscopy.
Carbon nanomembranes (CNMs) prepared from aromatic self-assembled monolayers constitute a recently developed class of 2D materials. They are made by a combination of self-assembly, radiation-induced cross-linking and the detachment of the cross-linked SAM from its substrate. CNMs can be deposited on arbitrary substrates, including holey and perforated ones, as well as on metallic (transmission electron microscopy) grids. Therewith, freestanding membranes with a thickness of 1 nm and macroscopic lateral dimensions can be prepared. Although free-standing CNMs cannot be imaged by light microscopy, charged particle techniques can visualize them. However, CNMs are electrically insulating, which makes them sensitive to charging. We demonstrate that the helium ion microscope (HIM) is a good candidate for imaging freestanding CNMs due to its efficient charge compensation tool. Scanning with a beam of helium ions while recording the emitted secondary electrons generates the HIM images. The advantages of HIM are high resolution, high surface sensitivity and large depth of field. The effects of sample charging, imaging of multilayer CNMs as well as imaging artefacts are discussed.


Cardiac Troponin I is Increased in Patients with Polytrauma and Chest or Head Trauma. Results of A Retrospective Case-Control Study.
We performed a retrospective case-control study to assess the values of cardiac troponin I (cTnI) in a large number of patients admitted to the emergency department (ED) with different types of trauma. The study population consisted of all patients aged 18 years or older admitted to the local ED with all types of traumas over a 1-year period. Results of cTnI were compared with those of 125 consecutive blood donors and 25 non-cardiac chest pain ED patients. The final study population consisted of 380 trauma patients, 10 with isolated abdominal trauma, 99 with isolated trauma of the limbs, 49 with isolated chest trauma, 145 with isolated head trauma and 77 with polytrauma. The concentration of cTnI did not differ among the three study populations, but the frequency of measurable values was substantially higher in patients with trauma (63%) than in blood donors and non-cardiac chest pain ED patients (both 20%). The frequency of cTnI values above the 99th percentile of the reference range was significantly higher in trauma patients (20%) than in blood donors (0%) and noncardiac chest pain ED patients (8%). Increased cTnI values were more frequent after head trauma (21%), chest trauma (27%) and polytrauma (29%) compared to patients with abdominal (0%) or limbs trauma (8%). These results suggest that the measurement of cardiac troponin may be advisable to identify potential cardiac involvement in trauma patients, especially in those with polytrauma and head or chest trauma.


[Clinical experience of endotracheal intubation after administration of NC45-10 (vecuronium bromide 10 mg preparation)].
We studied the usefulness of the 10 mg presentation of vecuronium bromide (NC45-10) for endotracheal intubation. Subjects were ASA type I-II patients between 20-65 years of age scheduled for various kinds of surgery who consented to participate in this study. After the entrance of patients into the operating room, we set up monitors and intravenously injected a priming dosage of NC45-10, 0.015 mg.kg-1. Next, we administered thiopental 5 mg.kg-1 and injected initial dosage of either 0.065 mg.kg-1 or 0.1 mg.kg-1 of NC45-10 intravenously 5 minutes after the administration of priming dose. One minute later, we attempted endotracheal intubation. We evaluated the muscle relaxation by the reaction to endotracheal intubation and graded it by the score of 0-3. Satisfactory muscle relaxation on endotracheal intubation was obtained in 80% of the patients with no significant difference between the 2 groups of initial dosage. No side effect or cardiovascular action was observed. It is concluded that this preparation is clinically safe and useful.


Potential autofertility in true hermaphrodites.
This article examines the studies on the pregnancies of true hermaphrodites and self-fertilization in hermaphrodite mammals that have been published in the last 40 years. The number of hermaphrodite pregnants reported in the literature since 1975 was 14, the number of pregnancies was 26 and the number of healthy born babies was 20. All of the babies that were born were male. The pregnancy developed following gonadectomy in seven cases (nine pregnancies). In some cases, either gonadectomy was not performed at all or it was performed after pregnancy (eight cases, 17 pregnancies). The karyotype was 46,XX in four of these eight cases that became pregnant despite in situ ovotestis while it was 46,XX/46,XY in the other four cases (chimera). In the literature, pregnancy cases that developed through self-fertilization were not reported in humans. However, autofertilization was detected in mammalian hermaphrodites such as domestic rabbit. [corrected]. Furthermore, the ovarian tissues of true hermaphrodites were mainly functional and ovulatory. The testicular tissues were mainly immature. However, spermatogenesis was determined in some cases. In fact, both ovulation and spermatogenesis were detected in some cases. All of these findings show that true hermaphrodites with ovarian and testicular tissues are potentially autofertile.


Granulosa cell-expressed BMPR1A and BMPR1B have unique functions in regulating fertility but act redundantly to suppress ovarian tumor development.
Bone morphogenetic proteins (BMPs) have diverse roles in development and reproduction. Although several BMPs are produced by oocytes, thecal cells, and granulosa cells of developing follicles, the in vivo functions of most of these ligands are unknown. BMP signals are transduced by multiple type I and type II TGFbeta family receptors, and of the type I receptors, BMP receptor 1A (BMPR1A) and BMP receptor 1B (BMPR1B) are known to be expressed in rodent granulosa cells. Female mice homozygous null for Bmpr1b are sterile due to compromised cumulus expansion, but the function of BMPR1A in the ovary is unknown. To further decipher a role for BMP signaling in mouse granulosa cells, we deleted Bmpr1a in the granulosa cells of the ovary and found Bmpr1a conditional knockout females to be subfertile with reduced spontaneous ovulation. To explore the redundant functions of BMP receptor signaling in the ovary, we generated Bmpr1a Bmpr1b double-mutant mice, which developed granulosa cell tumors that have evidence of increased TGFbeta and hedgehog signaling. Thus, similar to SMAD1 and SMAD5, which have redundant roles in suppressing granulosa cell tumor development in mice, two type I BMP receptors, BMPR1A and BMPR1B, function together to prevent ovarian tumorigenesis. These studies support a role for a functional BMP signaling axis as a tumor suppressor pathway in the ovary, with BMPR1A and BMPR1B acting downstream of BMP ligands and upstream of BMP receptor SMADs.


Development of a Low-Cost, Wireless Smart Thermostat for Isothermal DNA Amplification in Lab-On-A-Chip Devices.
Nucleic acid amplification tests (NAAT) are widely used for the detection of living organisms, recently applied in Lab-on-a-Chip (LoC) devices to make portable DNA analysis platforms. While portable LoC-NAAT can provide definitive test results on the spot, it requires specialized temperature control equipment. This work focuses on delivering a generalized low-cost, wireless smart thermostat for isothermal NAAT protocols in 2 cm × 3 cm LoC cartridges. We report on the design, prototyping, and evaluation results of our smart thermostat. The thermostat was evaluated by experimental and simulated thermal analysis using 3D printed LoC cartridges, in order to verify its applicability to various isothermal NAAT protocols. Furthermore, it was tested at the boundaries of its operating ambient temperature range as well as its battery life was evaluated. The prototype thermostat was proven functional in 20-30 °C ambient range, capable of maintaining the required reaction temperature of 12 isothermal NAAT protocols with 0.7 °C steady-state error in the worst case.


ALK1-Negative Anaplastic Large Cell Lymphoma of the Breast from a Nonprosthesis Cyst.
Anaplastic large cell lymphoma of the breast is a rare malignancy associated with prosthetic breast implants. We present a case of a woman with no prior history of breast implants who developed anaplastic lymphoma kinase-1 negative anaplastic large cell lymphoma on a background of a previous benign cyst aspiration.


Controlled surface plasmon enhanced fluorescence from 1D gold gratings via azimuth rotations.
We demonstrate a method to maximise the fluorescence enhancement from a dye using gold coated diffraction gratings. Rotations about the azimuth provides a convenient approach to maximise the coupling between the grating and excitation source while achieving enhancements comparable to traditional optical configurations where the grating and in plane light vectors are parallel. This approach yields a 30 fold enhancement in the fluorescence signal over metal free substrates, while opening up the range of possible orientations and configurations suitable for fluorescence enhancement applications.


Common therapeutic strategies for prion and Alzheimer's diseases.
A number of unexpected pathophysiological connections linking different neurodegenerative diseases have emerged over the past decade. An example is provided by prion and Alzheimer's diseases. Despite being distinct pathologies, these disorders share several neurotoxic mechanisms, including accumulation of misfolded protein isoforms, stress of the protein synthesis machinery, and activation of a neurotoxic signaling mediated by the cellular prion protein. Here, in addition to reviewing these mechanisms, we will discuss the potential therapeutic interventions for prion and Alzheimer's diseases that are arising from the comprehension of their common neurodegenerative pathways.


Distinct contributions of the fornix and inferior longitudinal fasciculus to episodic and semantic autobiographical memory.
Autobiographical memory (AM) is multifaceted, incorporating the vivid retrieval of contextual detail (episodic AM), together with semantic knowledge that infuses meaning and coherence into past events (semantic AM). While neuropsychological evidence highlights a role for the hippocampus and anterior temporal lobe (ATL) in episodic and semantic AM, respectively, it is unclear whether these constitute dissociable large-scale AM networks. We used high angular resolution diffusion-weighted imaging and constrained spherical deconvolution-based tractography to assess white matter microstructure in 27 healthy young adult participants who were asked to recall past experiences using word cues. Inter-individual variation in the microstructure of the fornix (the main hippocampal input/output pathway) related to the amount of episodic, but not semantic, detail in AMs - independent of memory age. Conversely, microstructure of the inferior longitudinal fasciculus, linking occipitotemporal regions with ATL, correlated with semantic, but not episodic, AMs. Further, these significant correlations remained when controlling for hippocampal and ATL grey matter volume, respectively. This striking correlational double dissociation supports the view that distinct, large-scale distributed brain circuits underpin context and concepts in AM.


[Comparison of total knee arthroplasty with computer navigation systems and conventional techniques].
To evaluate the value of total knee arthroplasty (TKA) with computer navigation by comparing with conventional TKA. Between May 2010 and December 2011, 45 patients underwent primary unilateral TKA, and the clinical data were retrospectively analyzed. Of 45 patients, 22 cases were treated with TKA with computer navigation (group A), 23 cases with the conventional TKA (group B). There was no significant difference in gender, age, body mass index, side, cause of disease, disease duration, preoperative range of motion (ROM) of the knee, and preoperative Hospital for Special Surgery (HSS) score between 2 groups (P > 0.05). The operation time, intraoperative blood loss, incidence of patellar retinacular release, complication, and drainage volume were compared. The prosthesis loosening, postoperative HSS score, and ROM of the knee were also compared. No difference was found in the incidence of patellar retinacular release during TKA, and it was 13.6% (3/22) in group A and was 4.3% (1/23) in group B, showing no significant difference (χ2 = 1.198, P = 0.346). The operation time of group A was significantly longer than that of group B (t = 7.557, P = 0.000). There was no significant difference in intraoperative blood loss during TKA between 2 groups (t = -0.295, P = 0.769), while the drainage volume of group A was significantly less than that of group B (t = -2.419, P = 0.020). Incomplete fracture during TKA and acute infection occurred at 8 days after TKA in 1 case of group A respectively, while no fracture or infection was found in group B, showing significant difference (Z = -0.509, P = 0.000). The patients of 2 groups were followed up 27-46 months. No significant difference in valgus and varus of knee, and malalignment of the femoral and tibial prosthesis was found (P > 0.05). There was no significant difference in HSS score and ROM of the knee at last follow-up between 2 groups (P > 0.05). No prosthesis loosening was found in 2 groups. TKA with computer navigation has similar results to conventional TKA in the mechanical alignment, but it obviously prolongs operation time. It may also increase the incidence of infection and tractor pin related fracture.


An integrative functional genomics framework for effective identification of novel regulatory variants in genome-phenome studies.
Genome-phenome studies have identified thousands of variants that are statistically associated with disease or traits; however, their functional roles are largely unclear. A comprehensive investigation of regulatory mechanisms and the gene regulatory networks between phenome-wide association study (PheWAS) and genome-wide association study (GWAS) is needed to identify novel regulatory variants contributing to risk for human diseases. In this study, we developed an integrative functional genomics framework that maps 215,107 significant single nucleotide polymorphism (SNP) traits generated from the PheWAS Catalog and 28,870 genome-wide significant SNP traits collected from the GWAS Catalog into a global human genome regulatory map via incorporating various functional annotation data, including transcription factor (TF)-based motifs, promoters, enhancers, and expression quantitative trait loci (eQTLs) generated from four major functional genomics databases: FANTOM5, ENCODE, NIH Roadmap, and Genotype-Tissue Expression (GTEx). In addition, we performed a tissue-specific regulatory circuit analysis through the integration of the identified regulatory variants and tissue-specific gene expression profiles in 7051 samples across 32 tissues from GTEx. We found that the disease-associated loci in both the PheWAS and GWAS Catalogs were significantly enriched with functional SNPs. The integration of functional annotations significantly improved the power of detecting novel associations in PheWAS, through which we found a number of functional associations with strong regulatory evidence in the PheWAS Catalog. Finally, we constructed tissue-specific regulatory circuits for several complex traits: mental diseases, autoimmune diseases, and cancer, via exploring tissue-specific TF-promoter/enhancer-target gene interaction networks. We uncovered several promising tissue-specific regulatory TFs or genes for Alzheimer's disease (e.g. ZIC1 and STX1B) and asthma (e.g. CSF3 and IL1RL1). This study offers powerful tools for exploring the functional consequences of variants generated from genome-phenome association studies in terms of their mechanisms on affecting multiple complex diseases and traits.


Aberrant V(D)J cleavages in T cell receptor beta enhancer- and p53-deficient lymphoma cells.
Previously, we generated thymic lymphoma cell lines from EbetaR/Rp53-/- (EP) double mutant mice where the T cell receptor (TCR) beta enhancer (Ebeta) was deleted, and the p53 gene was inactivated. Here, we characterized the EP cell lines to study the roles of the Ebeta and p53 on TCRbeta rearrangements during lymphomagenesis. Recombination activation genes (RAGs) were expressed, while the TCRbeta chain was not expressed in the EP cell lines. Dbeta-Jbeta rearrangements were not detected at all, and Dbeta1 and Dbeta2 cleavages were also not detected in the EP cell lines. However, Jbeta cleavages suppressed in Ebeta mutant thymocytes were readily detected in the EP cell lines. The Jbeta cleavages appeared to be uncoupled, aberrant, RAG-dependent and Ebeta-independent and were not detected in a p53 or Ebeta single mutant background, suggesting that the Jbeta cleavages are selected in the Ebeta and p53 double mutant background. Sequence analysis showed that the cleavage occurred in the cryptic recombination signal sequences (RSSs) present throughout Jbeta gene segments. The results implicate that the uncoupled and aberrant V(D)J cleavages may contribute to double-strand break-mediated genome instability during lymphomagenesis in EP mice.


Regulated expression and function of CD11c/CD18 integrin on human B lymphocytes. Relation between attachment to fibrinogen and triggering of proliferation through CD11c/CD18.
CD11c/CD18 (p150,95) is a beta 2 integrin expressed by myeloid, natural killer and certain lymphoid cells such as some cytotoxic T cell clones and B cell malignancies. We have studied the expression and function of CD11c on resting and activated B lymphocytes. Flow cytometry, immunoprecipitation, and mRNA analyses showed that cell activation with phorbol esters or with a variety of stimuli such as Staphylococcus aureus or anti-mu antibodies in combination with cytokines induced de novo CD11c/CD18 cell surface expression on most B cells while CD11b expression was not affected. Functional analysis of CD11c/CD18 on B cells revealed that it plays a dual role. First, CD11c/CD18 is implicated in B cell proliferation, as demonstrated by the ability of several anti-CD11c monoclonal antibodies to trigger comitogenic signals; and second, the newly expressed CD11c/CD18 mediates B cell binding to fibrinogen. Our data conclusively demonstrate the role of CD11c/CD18 on both B cell activation and adhesion processes.