Autophagy and radiosensitization in cancer.
Autophagy is a natural self-degradative process by which cells eliminate misfolded proteins and damaged organelles. Autophagy has been shown to have multiple functions in tumor cells that may be dependent on the tumor type and the treatment conditions. Autophagy can have a cytoprotective role and be thought of as a survival mechanism or be cytotoxic in nature and mediate cell death. Radiation, one of the primary treatments for many different types of cancer, almost uniformly promotes autophagy in tumor cells. While autophagy produced in response to radiation is often considered to be cytoprotective, radiation-induced autophagy has also been shown to mediate susceptibility to radiation. This review addresses the complexity of autophagy in response to radiation treatment in three different cancer models, specifically lung cancer, breast cancer and glioblastoma. A deeper understanding of the different roles played by autophagy in response to radiation should facilitate the development of approaches for enhancing the therapeutic utility of radiation by providing strategies for combination treatment with unique radiosensitizers as well as preventing the initiation of strategies which are likely to attenuate the effectiveness of radiation therapy.


Phase separation of equilibrium polymers of proteins in living cells.
A number of proteins polymerise reversibly in living cells. The equilibrium polymers are functional: if mutant proteins are made that cannot polymerise, then these proteins cannot perform their biological functions. Furthermore, these polymers of proteins appear to phase separate inside the cell. The dynamics of one of these polymerising, phase separating proteins has been studied via fluorescence recovery after photobleaching (FRAP) by Bienz and coworkers. Here, their data is compared with the results of quantitative modelling to gain a better understanding of the dynamics of this protein inside a cell. The protein is called Dishevelled; it is a protein essential to the development of all animals and the name originates in the disruption of hair formation in a mutant version of this protein. It is not known how polymerisation and phase separation enable Dishevelled to perform its biological function but here we propose and discuss two possibilities. The first is that the cell is exploiting the inherently sharp, switch-like nature of a phase transition to respond in a switch-like way to a external signal. The second is that phase separation dynamically creates a compartment (the more concentrated phase) into which other proteins partition.


Interleukin 2 promotes expression of an interleukin 2 receptor and proliferation of T cells stimulated with non-mitogenic dose of concanavalin A.
Interleukin 2 (IL-2) initiated proliferation of the cells of T-enriched population stimulated with non-mitogenic dose of concanavalin A (Con A), whereas it could not induce proliferation of the cells treated with non-mitogenic lectin wheat germ agglutinin (WGA). Use of monoclonal antibody (MAb) directed against interleukin 2 receptor (IL-2R) showed that IL-2 significantly increased expression of IL-2R on the cells of T-enriched population stimulated with Con A, whereas it could not cause any significant enhancement of expression of IL-2R on the cells treated with WGA. We concluded that IL-2 activated T cells in combination with non-mitogenic does of Con A, and induced both expression of IL-2R and proliferation.


Role of mental health professionals in the management of functional somatic symptoms in primary care.
Functional somatic symptoms associated with persistent frequent attendance is emotionally demanding, costly and intractable to treat. Such patients are hard to engage in practice and research by mental health professionals, whose main role may be indirect training, supporting and advising primary care professionals rather than direct patient care.


The effects of propofol anaesthesia on molecular-enriched networks during resting-state and naturalistic listening.
Placing a patient in a state of anaesthesia is crucial for modern surgical practice. However, the mechanisms by which anaesthetic drugs, such as propofol, impart their effects on consciousness remain poorly understood. Propofol potentiates GABAergic transmission, which purportedly has direct actions on cortex as well as indirect actions via ascending neuromodulatory systems. Functional imaging studies to date have been limited in their ability to unravel how these effects on neurotransmission impact the system-level dynamics of the brain. Here, we leveraged advances in multi-modal imaging, Receptor-Enriched Analysis of functional Connectivity by Targets (REACT), to investigate how different levels of propofol-induced sedation alter neurotransmission-related functional connectivity (FC), both at rest and when individuals are exposed to naturalistic auditory stimulation. Propofol increased GABA-A- and noradrenaline transporter-enriched FC within occipital and somatosensory regions respectively. Additionally, during auditory stimulation, the network related to the dopamine transporter showed reduced FC within bilateral regions of temporal and mid/posterior cingulate cortices, with the right temporal cluster showing an interaction between auditory stimulation and level of consciousness. In bringing together these micro- and macro-scale systems, we provide support for both direct GABAergic and indirect noradrenergic and dopaminergic-related network changes under propofol sedation. Further, we delineate a cognition-related reconfiguration of the dopaminergic network, highlighting the utility of REACT to explore the molecular substrates of consciousness and cognition.


miRNAs as biofluid markers for diagnostics of Alzheimer's disease: recent status and perspectives.
After many decades of research in the field of neurodegeneration, we have no effective cure for Alzheimer's disease (AD), a major form of dementia. It is mainly due to the lack of early, reliable and sensitive biomarkers and incomplete understanding of disease mechanisms at molecular level. Several recently employed biomarkers, especially their combinations, can discriminate advanced stages of AD from other forms of dementia or neuropathy. They do not provide much information on molecular mechanisms of disease rather they reflect the amount of key histopathological markers in the diseased brain. This review is focussed on novel class of potentially very promising AD biomarkers: extracellular miRNAs in body liquids, such as cerebrospinal fluid and blood. They have a great potential not only to indicate the presence of AD, but more importantly, to reflect the molecular mechanisms playing a role early at the beginning of the pathogenic pathways consequently leading to AD. We believe this comprehensive review on deregulated miRNAs in AD can be a good source of information for thorough in silico analyses aiming to identification, development and validation of miRNAs as "diseases mechanism engaged" candidate biomarkers. Having such molecules could bring us closer to the goal - successful treatment of AD.


The cation-π interaction between Lys53 and the flavin of fructosamine oxidase (FAOX-II) is critical for activity.
Fructosamine oxidases (FAOXs) are flavin-containing enzymes that catalyze the oxidative deglycation of low molecular weight fructosamines or Amadori products. The fructosamine substrate is oxidized by the flavin in the reductive half-reaction, and the reduced flavin is then oxidized by molecular oxygen in the oxidative half-reaction. The crystal structure of FAOX-II from Aspergillus fumigatus reveals a unique interaction between Lys53 and the isoalloxazine. The ammonium nitrogen of the lysine is in contact with and nearly centered over the aromatic ring of the flavin on the si-face. Here, we investigate the importance of this unique interaction on the reactions catalyzed by FAOX by studying both half-reactions of the wild-type and Lys53 mutant enzymes. The positive charge of Lys53 is critical for flavin reduction but plays very little role in the reaction with molecular oxygen. The conservative mutation of Lys53 to arginine had minor effects on catalysis. However, removing the charge by replacing Lys53 with methionine caused more than a million-fold decrease in flavin reduction, while only slowing the oxygen reaction by ∼30-fold.


Incorporating evolutionary processes into population viability models.
We examined how ecological and evolutionary (eco-evo) processes in population dynamics could be better integrated into population viability analysis (PVA). Complementary advances in computation and population genomics can be combined into an eco-evo PVA to offer powerful new approaches to understand the influence of evolutionary processes on population persistence. We developed the mechanistic basis of an eco-evo PVA using individual-based models with individual-level genotype tracking and dynamic genotype-phenotype mapping to model emergent population-level effects, such as local adaptation and genetic rescue. We then outline how genomics can allow or improve parameter estimation for PVA models by providing genotypic information at large numbers of loci for neutral and functional genome regions. As climate change and other threatening processes increase in rate and scale, eco-evo PVAs will become essential research tools to evaluate the effects of adaptive potential, evolutionary rescue, and locally adapted traits on persistence.


The altered balance between sympathetic nervous system and nitric oxide in salt hypertensive Dahl rats: ontogenetic and F2 hybrid studies.
We have demonstrated earlier that the nitric oxide (NO) system is not able to counterbalance effectively the hyperactivity of the sympathetic nervous system (SNS) in salt hypertension of young Dahl rats in which augmented superoxide anion formation lowers NO bioavailability. The aim of the present study was to determine whether SNS hyperactivity and/or relative NO deficiency are also present in salt hypertension elicited in adult Dahl rats, and whether they are associated with blood pressure (BP) in the F2 population of Dahl rats. The contribution of major vasoactive systems [renin-angiotensin system (RAS), SNS and NO] and superoxide anions to BP maintenance was studied in SS/Jr rats in which salt hypertension was induced either in adulthood or in youth (8% NaCl diet from the age of 12 or 4 weeks). The contribution of particular vasoactive systems was also investigated in 122 young salt-loaded F2hybrids [derived from salt-sensitive (SS/Jr) and salt-resistant (SR/Jr) Dahl rats] which were fed a high-salt diet (8% NaCl) for 6 weeks after weaning. Mean arterial pressure (MAP) was measured in conscious animals subjected to acute consecutive blockade of RAS (captopril 10 mg/kg i.v.), SNS (pentolinium 5 mg/kg i.v.) and NO synthase (l-NAME 30 mg/kg i.v.). Dahl rats with salt hypertension induced in adulthood were also characterized by enhanced pentolinium-induced BP fall (DeltaMAPpento), but their residual BP (recorded after the blockade of both RAS and SNS) was unaltered, in contrast to its elevation seen in young salt-hypertensive rats. The BP rise after NO synthase inhibition by l-NAME (DeltaMAPL-NAME), which was substantially greater in adult than in young hypertensive rats, was not enhanced by superoxide scavenging with tempol in adult hypertensive animals, in which this drug elicited a moderate BP reduction only. Basal MAP of young salt-loaded F2 hybrids was positively associated not only with DeltaMAPpento (P < 0.0001) and residual BP (P < 0.001) but also with DeltaMAPL-NAME (P < 0.001). The slope of the relationship between basal BP and pentolinium-induced BP changes was steeper than that between basal BP and BP changes elicited by l-NAME. The positive correlation of basal BP with DeltaMAPpento/DeltaMAPL-NAME ratio (P < 0.01) indicates that an altered balance between sympathetic vasoconstriction and NO-dependent vasodilation was associated with high blood pressure, even in the F2 population of Dahl rats. A comparison of young and adult salt-hypertensive Dahl rats stressed the importance of increased residual BP and relative NO deficiency for the severity of hypertension, because these two alterations were absent in a less-pronounced form of salt hypertension elicited in adulthood. The findings obtained in our young salt-loaded F2 population also confirm the major importance of both sympathetic hyperactivity and relative NO deficiency for the maintenance of salt hypertension in Dahl rats.


Focal Breast Lesions in Clinical CT Examinations of the Chest: A Retrospective Analysis.
Purpose Based on radiological reports, the percentage of breast cancers visualized as incidental findings in routine CT examinations is estimated at ≤ 2 %. In view of the rising number of CT examinations and the high prevalence of breast cancer, it was the goal of the present study to verify the frequency and image morphology of false-negative senological CT findings. Materials and Methods All first contrast-enhanced CT examinations of the chest in adult female patients carried out in 2012 were retrospectively included. A senior radiologist systematically assessed the presence of breast lesions on all CT images using the BI-RADS system. All BI-RADS ≥ 3 notations were evaluated by a second senior radiologist. A consensus was obtained in case of differing BI-RADS assessments. Reference diagnoses were elaborated based on all available clinical, radiological and pathological data. The findings of the CT reports were classified according to the BI-RADS system and were compared with the retrospective consensus findings as well as with the reference diagnoses. Results The range of indications comprised a broad spectrum including staging and follow-up examinations of solid tumors/lymphoma (N = 701, 59.9 %) and vascular (190, 16.2 %), inflammatory (48, 4.1 %) and pulmonologic (22, 1.9 %) issues. BI-RADS 1/2 classifications were present in 92.5 % and BI-RADS 6 classifications were assessed in 1.7 % of the 1170 included examinations. 68 patients (5.8 %) had at least one lesion retrospectively classified as BI-RADS 3 - 5. The histological potential was known in 57 of these lesions as benign (46, 3.9 %) or malignant (11, 0.9 %). 13 BI-RADS 4/5 consensus assessments (1.1 %) were false-positive. 2 of the 10 lesions classified as being malignant based on the further clinical and radiological course were not mentioned in the written CT reports (0.2 %). Both false-negative CT reports were therapeutically and prognostically irrelevant. Conclusion The relative frequency of BI-RADS 3 - 5 findings was 5.8 %. It reflects the situation encountered in clinical imaging for primarily non-senologic questions and therefore differs from what would be expected in a dedicated screening program. The rates of known false-positive BI-RADS 4/5 findings in the retrospective evaluations (1.1 %) and of false-negative findings in the written CT reports (0.2 %) reflect the different diagnostic approaches of image-based senological screening and radiological examinations indicated in order to solve clinical problems not primarily concerning the breast region. Statements regarding the prevalence of clinically occult breast cancers can only be made with caution in the presented, highly selective group of patients due to the often incomplete visualization of breast tissue and the retrospective approach. Key points · Intramammary mass and non-mass lesions needing clarification may be present in up to 5.8 % of all contrast enhanced CT-examinations of the female chest.. · Irregular forms, unscharp/spiculated margins, inhomogeneous matrices and a pronounced contrast medium enhancement point towards a malignant genesis of an intramammary mass or non-mass lesion.. · The results of the study highlight the importance of paying systematical and targeted attention on senological additional findings in CT-examinations of the chest also in other clinical settings than that of the included patients in a clinic with oncological main focus.. Zitierweise · Krug KB, Houbois C, Grinstein O et al. Focal Breast Lesions in Clinical CT Examinations of the Chest: A Retrospective Analysis. Fortschr Röntgenstr 2017; 189: 977 - 988.


Topical application of nerve growth factor improves fracture healing in rats.
The aim of the present study was to examine the effects of nerve growth factor on the healing of unsplinted fractured ribs. After fracture of a rib in male rats, nerve growth factor was delivered by a miniosmotic pump to the fracture site for 7 days at the rate of 1.4 micrograms/day. Callus catecholamine concentrations, bone callus size, histomorphometry, and biomechanical properties of the repairing rib were measured at 7, 21, and 42 days after fracture. After 21 days, concentrations of norepinephrine and epinephrine were significantly increased in the group treated with nerve growth factor compared with those in the control group (211% norepinephrine and 322% epinephrine). Also, the midline longitudinal area of non-osseous (fibrous tissue and cartilage) callus of the fracture was significantly smaller (54%) and had a higher proportion of cartilage in the treated group than in the controls. By 42 days, there was only bony callus between the fracture ends in both the control group and the treated group. The treated group, however, again showed significantly elevated concentrations of norepinephrine and epinephrine (286 and 382%, respectively) and significantly elevated breaking stress (50%) and Young's modulus (51%), together with a reduction in the transverse cross-sectional area of the repair site (57%). The resultant increases in effectiveness and rate of repair of bone with administration of nerve growth factor suggest that it may play an important role in the healing processes of fractured bone.


Radiation and electrostatic resistance for ultra-stable polymer composites reinforced with carbon fibers.
Future space travel needs ultra-lightweight and robust structural materials that can withstand extreme conditions with multiple entry points to orbit to ensure mission reliability. This is unattainable with current inorganic materials. Ultra-highly stable carbon fiber reinforced polymers (CFRPs) have shown susceptibility to environmental instabilities and electrostatic discharge, thereby limiting the full lightweight potential of CFRP. A more robust and improved CFRP is needed in order to improve space travel and structural engineering further. Here, we address these challenges and present a superlattice nano-barrier-enhanced CFRP with a density of ~3.18 g/cm3 that blends within the mechanical properties of the CFRP, thus becoming part of the composite itself. We demonstrate composites with enhanced radiation resistance coupled with electrical conductivity (3.2 × 10-8 ohm⋅m), while ensuring ultra-dimensionally stable physical properties even after temperature cycles from 77 to 573 K.


Matrix metalloproteinase-9 and -7 are regulated in experimental autoimmune encephalomyelitis.
Matrix metalloproteinases (MMPs) comprise a group of proteolytic enzymes that are implicated in the pathogenesis of inflammatory diseases of the nervous system such as multiple sclerosis. However, the exact function and expression pattern of MMPs in the inflamed nervous system are not known. In the present study we investigated the expression of 92-kDa gelatinase (MMP-9) in spinal cord from animals with adoptive transfer experimental autoimmune encephalomyelitis (AT-EAE), using a semiquantitative competitive reverse transcriptase-polymerase chain reaction assay. Increased levels of MMP-9 mRNA were found with peak values at times of maximum disease severity. Increased mRNA expression was associated with enhanced proteolytic activity of this enzyme, as demonstrated by gelatin zymography. Immunohistochemistry revealed immunoreactivity along the meninges, around blood vessels and within the parenchyma, in diseased but not in normal spinal cord. Furthermore, the expression pattern of five other MMPs was investigated. Matrilysin (MMP-7) was also found to be upregulated with maximum mRNA levels at the peak of the disease. In contrast, mRNAs for collagenase-3, 72-kDa gelatinase, and stromelysin-1 and -3 were not changed. Our findings indicate that 92-kDa gelatinase and matrilysin are selectively upregulated during AT-EAE and thus may contribute to the pathogenesis of inflammatory diseases of the CNS.


High dose naloxone in depression.
The behavioral effects of a 2 mg/kg iv bolus infusion of naloxone were compared with a placebo infusion using a double-blind design in a small group of inpatient depressives (n = 6) and normals (n = 8). Naloxone produced consistent and significant worsening in the rated signs and subjective symptoms of depression in the patients. In the normals, lesser changes in Hamilton depression and BPRS total scores were observed while none of the subjective scales were significantly altered. The data suggest that depressives manifest a more marked and subjectively more intense response to naloxone compared to normals. Further studies are required to confirm this preliminary finding and to clarify its relationship to the pathogenesis of depression.


Long Non-coding RNA TMEM220-AS1 Suppressed Hepatocellular Carcinoma by Regulating the miR-484/MAGI1 Axis as a Competing Endogenous RNA.
Long non-coding RNAs (lncRNAs) have a considerable regulatory influence on multiple biological processes. Nevertheless, the role of TMEM220-AS1 in hepatocellular carcinoma (HCC) remains unclear. We used The Cancer Genome Atlas (TCGA) database to analyze the differentially expressed lncRNAs. qRT-PCR was used to verify the results for a large population. The in vitro effects of TMEM220-AS1 on HCC cells were determined using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, and Transwell assays in HCC cells. We used qRT-PCR and western blotting to identify the epithelial-mesenchymal transition (EMT). Moreover, we performed bioinformatics analysis, western blotting, dual luciferase reporter gene assay, RNA pull-down, and RNA binding protein immunoprecipitation (RIP) to investigate the underlying molecular mechanisms of TMEM220-AS1 function. Finally, the function of TMEM220-AS1 was verified in vivo. The results showed that TMEM220-AS1 was expressed at considerably low levels in HCC. It was demonstrated that malignant phenotypes and EMT of HCC cells were promoted by the knock down of TMEM220-AS1 both in vivo and in vitro. TMEM220-AS1, which was detected primarily in the cytoplasm, functioned as an miRNA sponge to bind miR-484 and promote the level of membrane-associated guanylate kinase, WW, and PDZ domain containing 1 (MAGI1), thereby curbing the malignant phenotypes of HCC cells. In conclusion, low levels of TMEM220-AS1 promote proliferation and metastasis through the miR-484/MAGI1 axis in HCC.


Genetic alterations in thyroid cancer: the role of mouse models.
Thyroid carcinomas are the most common endocrine neoplasms in humans, with a globally increasing incidence. Thyroid follicular cells and neuroendocrine (parafollicular) C cells are each susceptible to neoplastic transformation, resulting in thyroid cancers of differing phenotypes with unique associated genetic mutations and clinical outcomes. Over the past 15 years, several sophisticated genetically engineered mouse models of thyroid cancer have been created to further our understanding of the genetic events leading to thyroid carcinogenesis in vivo. The most significant mouse models of papillary, follicular, anaplastic, and medullary thyroid carcinoma are highlighted, with particular emphasis on the relationship between the relevant oncogenes in these models and genetic events in the naturally occurring human disease. Limitations of each model are presented, and the need for additional models to better recapitulate certain aspects of the human disease is discussed.


How do the smart travel ban policy and intercity travel pattern affect COVID-19 trends? Lessons learned from Iran.
COVID-19, as the most significant epidemic of the century, infected 467 million people and took the lives of more than 6 million individuals as of March 19, 2022. Due to the rapid transmission of the disease and the lack of definitive treatment, countries have employed nonpharmaceutical interventions. This study aimed to investigate the effectiveness of the smart travel ban policy, which has been implemented for non-commercial vehicles in the intercity highways of Iran since November 21, 2020. The other goal was to suggest efficient COVID-19 forecasting tools and to examine the association of intercity travel patterns and COVID-19 trends in Iran. To this end, weekly confirmed cases and deaths due to COVID-19 and the intercity traffic flow reported by loop detectors were aggregated at the country's level. The Box-Jenkins methodology was employed to evaluate the policy's effectiveness, using the interrupted time series analysis. The results indicated that the autoregressive integrated moving average with explanatory variable (ARIMAX) model outperformed the univariate ARIMA model in predicting the disease trends based on the MAPE criterion. The weekly intercity traffic and its lagged variables were entered as covariates in both models of the disease cases and deaths. The results indicated that the weekly intercity traffic increases the new weekly COVID-19 cases and deaths with a time lag of two and five weeks, respectively. Besides, the interrupted time series analysis indicated that the smart travel ban policy had decreased intercity travel by around 29%. Nonetheless, it had no significant direct effect on COVID-19 trends. This study suggests that the travel ban policy would not be efficient lonely unless it is coupled with active measures and adherence to health protocols by the people.


Influence of coagulation factors on intrinsic thrombin generation.
The intrinsic coagulation activity assay (INCA) is a new thrombin-generation test that imitates the intrinsic pathway of blood coagulation. The aim of the present study was to investigate the influence of the main coagulation factors on the INCA. The INCA was performed with citrated plasma samples supplemented with different amounts of fibrinogen. The INCA and activated partial thromboplastin time determination were performed with factor-depleted plasmas and with mixtures of depleted plasmas with normal plasma. Supplemented purified fibrinogen resulted in a decrease of intrinsic thrombin generation (50% inhibitory concentration = 0.8 g/l). The INCA depends on the intrinsic factors (factors VIII, IX, XI and XII) and on the factors of the common pathway (factors II, V and X): for normal thrombin generation, at least about 50% of normal factor II is necessary. For the majority of factors, the sensitivity of the INCA appears to be approximately one order of magnitude better than that of the activated partial thromboplastin time. The INCA allows one to diagnose defects in the intrinsic coagulation system and might be a useful test to support development and characterization of new drugs targeted at the intrinsic generation of thrombin.


Three-dimensional echocardiography in congenital malformations of the mitral valve.
Three-dimensional echocardiography has proved to be valuable in congenital heart disease by enhancing the evaluation of morphologic abnormalities and increasing the understanding of complex relationships. This study was undertaken to determine how 3-dimensional echocardiography could be best used to study some of the congenital malformations of the mitral valve such as mitral arcade, double orifice mitral valve, accessory mitral tissue, cleft mitral valve, and unicuspid mitral valve. Five patients were studied. Three-dimensional echocardiography was found to be helpful in defining spatial location and extent of deformities.


WBP2 Downregulation Inhibits Proliferation by Blocking YAP Transcription and the EGFR/PI3K/Akt Signaling Pathway in Triple Negative Breast Cancer.
Dysregulated expression of WW domain-binding protein 2 (WBP2) is associated with poor prognosis in ER+ breast cancer patients. However, its role in triple negative breast cancer (TNBC) has not been previously assessed. Therefore, we aimed to elucidate the functional mechanism of WBP2 in TNBC cells. qRT-PCR, western blotting, and immunohistochemical staining were used to evaluate WBP2 expression in TNBC patient tumors and cell lines. HCC1937 and MDA-MB-231 cells transiently transfected with WBP2 small interfering RNA (siRNA), miR-613 mimics, or miR-613 inhibitors were subject to assays for cell viability, apoptosis and cell cycle distribution. Co-immunoprecipitation, western blotting or qRT-PCR were employed to monitor changes in signaling pathway-related genes and proteins. Luciferase assays were performed to assess whether WBP2 is a direct target of miR-613. The effect of miR-613 on tumor growth was assessed in vivo using mouse xenograft models. The expression of WBP2 was upregulated in TNBC tissues and cells. Expression of WBP2 was significantly correlated with Ki67 in TNBC patients. Knockdown of WBP2 inhibited cellular proliferation, promoted apoptosis, and induced cell cycle arrest of TNBC cells. miR-613 directly bound to the 3'-untranslated region (3'-UTR) of WBP2 and regulated the expression of WBP2. Moreover, miR-613 reduced the expression of WBP2 and suppressed tumor growth of TNBC cells in vivo. Knockdown of WBP2 inhibited YAP transcription and the EGFR/PI3K/Akt signaling pathway in TNBC cells, and these effects were reversed by inhibition of miR-613. WBP2 overexpression is associated with the poor prognosis of TNBC patients and the miR-613-WBP2 axis represses TNBC cell growth by inactivating YAP-mediated gene expression and the EGFR/PI3K/Akt signaling pathway.


Topical application of decongestant in dysfunction of the Eustachian tube: a randomized, double-blind, placebo-controlled trial.
Thirty-six patients, aged 12-75 years, with dry, central tympanic membrane perforations and a negative Valsalva manouevre and/or a negative aspiration/deflation test, were included in a randomized, double-blind, placebo-controlled trial on the effect of a decongestant agent (xylometazoline chloride 0.1%) and placebo (saline 0.9%) applied directly to the pharyngeal opening of the Eustachian tube. Judged by the Valsalva manouevre, tubal patency was significantly improved after application of the active drug (P less than 0.003). In contrast, no effect was demonstrated by the aspiration test (P = 0.80) or the deflation test (P = 0.51). It is concluded that a topical decongestant improves Eustachian tube function but only at unphysiologically high pressures.


Sonography of the kidney and urinary tract.
Sonography has become integral to the evaluation and management of patients with renal disease. Although many of the procedures discussed in this issue are not feasible for all nephrologists, sonography clearly is. It is convenient and saves time, both for the physician and the patient, and the equipment is not expensive. It is not a difficult skill to learn, as evidenced by the growing number of nephrologists who have learned it, and it greatly enhances the practice of nephrology. Even nephrologists who do not plan to acquire this skill should be familiar with it to understand the indications and limitations, and to participate in its interpretation. Sonography, both the performance and interpretation, is an acquired skill that requires hands-on training, supervision, and practice, and thus cannot be learned solely by reading. Therefore, the goal here is not to teach sonography but rather to provide a basic overview of sonography, discuss its role in nephrology, and present information relevant to incorporating sonography into a nephrology practice.


A test of magnitude: does the strength of predictors explain differences in drug use among adolescents?
Prior research has provided much needed information on the association between gender, race, predictive factors, and drug use. However, none have provided conclusive evidence regarding the extent to which gender and race differences, or their interaction, in adolescent drug use is accounted for by predictors. Using data on adolescents in four public schools, this study examined variations in the comparative strength of predictors of self-reported drug use. Data were collected as part of an ongoing longitudinal study of a federally funded multi-model program intervention for the prevention of alcohol and drug abuse among high risk youth. Analysis of variance was used to examine gender by race differences in predictors and drug use and logistic regression was used to assess the comparative strength of predictors of adolescent drug use and to explore the effect of gender by race on the decision to use drugs. Findings indicated, in general, whites, both male and female, had a significantly stronger positive association between predictors and drug use relative to blacks. Regarding the effect of gender by race on the decision to use drugs, these data indicate, with or without adjustments, white females were most likely to use gateway substances, followed by white males.


Current state of medical device nomenclature and taxonomy systems in the UK: spotlight on GMDN and SNOMED CT.
A standardised terminology for describing medical devices can enable safe and unambiguous exchange of information. Proposed changes to EU-wide medical devices regulations mandate the use of such a system. This article reviews two important classification systems for medical devices in the UK. The Global Medical Device Nomenclature (GMDN) provides a classification system specifically for medical devices and diagnostics, and facilitates data exchange between manufacturers and regulators. SNOMED CT is the terminology of choice in the NHS for communicating, sharing and storing information about patients' healthcare episodes. Harmonisation of GMDN and SNOMED CT will encourage use of single terminology throughout the lifetime of a device; from regulatory approval through clinical use and post-marketing surveillance. Manufacturers will be required to register medical devices with a European device database (Eudamed) and to fit certain devices with a Unique Device Identifier; both are efforts to improve transparency and traceability of medical devices. Successful implementation of these elements depends on having a consistent nomenclature for medical devices.


Age-related IQ decline is reduced markedly after adjustment for the Flynn effect.
Twenty-year-olds outperform 70-year-olds by as much as 2.3 standard deviations (35 IQ points) on subtests of the Wechsler Adult Intelligence Scale (WAIS). We show that most of the difference can be attributed to an intergenerational rise in IQ known as the Flynn effect. Normative data from different versions of the WAIS enabled us to estimate the degree to which the Flynn effect, rather than age-related decline, contributes to differences between 20- and 70-year-olds. The Flynn effect accounted for 38-67% of the apparent age-related decline on 6 of the 11 subtests. On the other 5 subtests, all of which are categorized as verbal, the Flynn effect was larger than the age-group difference. For these verbal subtests, the Flynn effect masked a modest increase in ability as individuals grow older. Overall, the Flynn effect accounted for at least 85% of the disparity between 20- and 70-year-olds.


The human interferon alpha-receptor protein confers differential responses to human interferon-beta versus interferon-alpha subtypes in mouse and hamster cell transfectants.
The human interferon alpha-receptor (IFNAR gene product or IFN alpha R protein) was expressed in hamster CHO cells and in mouse A9 cells. The response of the IFN alpha R cDNA transfectants to human IFNs was studied by measuring induction of (2'-5') A synthetase (2'-5' AS). In the murine cells, the IFN alpha R protein conferred response to the human IFN-alpha-8 (alpha-B) subtype, but not to huIFN-alpha-2 (alpha-A) or to huIFN-beta. In murine huIFN alpha R cDNA transfectants, containing a hygromycin B resistance gene placed under the control of the 2'-5' AS gene Interferon Response Sequence (IRS), survival and growth of the cells in the presence of hygromycin B was induced by huIFN-alpha-8 but not by huIFN-alpha-2, indicating that the effect of huIFN alpha R is transcriptional. In hamster CHO cells, the huIFN alpha R protein conferred a completely different pattern of response to human IFN subtypes. Thus, the CHO-IFN alpha R transfectants responded to huIFN-beta by 2'-5' AS induction as well as by activation of the ISGF3 and IRF-1 transcription factors. In contrast, the CHO-IFN alpha R cells showed no response to huIFN-alpha-8. The differential response conferred by the huIFN alpha R protein in the two types of rodent cells, indicates that IFN subtype recognition is influenced by another component contributed by the rodent host cell. The ability of human cells, and of human-mouse hybrid cells containing human chromosome 21, to respond to all IFN subtypes is likely to depend also on interactions of the IFN alpha R protein with additional receptor components.


Surface functionalized mesoporous silica nanoparticles as an effective carrier for epirubicin delivery to cancer cells.
Recent studies with inorganic nanoparticles modified with functional groups have demonstrated improvement in drug delivery into cancer cells. In the present study, we prepared, characterized, and evaluated mesoporous silica nanoparticles (MSNs) as carriers for epirubicin hydrochloride (EPI) in order to improve the antitumor efficacy of this drug. MSNs were prepared and functionalized with phosphonate, polyethylene glycol (PEG) and polyethylenimine-polyethylene glycol (PEI-PEG) groups. Different nanoparticulate formulations were loaded with EPI. The in vitro cytotoxicity and the in vivo antitumor efficacy of MSNs containing EPI were evaluated versus free EPI. The EPI release from nanoparticles was shown to be pH-dependent. The size of MSNs functionalized with polyethyleneimine-polyethylene glycol (MSN-PEI-PEG) was 123.8 ± 4.8 nm. This formulation showed the best antitumor effects at an EPI dose of 9 mg/kg in C-26 colon carcinoma model. The biodistribution results proved that MSN-PEI-PEG-EPI had a higher tumor accumulation compared to free EPI, 3h after drug administration. The results indicated that this formulation could be effective nanocarriers for anti-tumor therapies.


Polysialic acid affects pathophysiological consequences of status epilepticus.
Modulation of the neural cell adhesion molecule by the attachment of polysialic acid residues through the polysialyl-transferase, ST8SiaIV, regulates neuronal plasticity and affects cellular alterations in the epileptic brain. Here, we determined the impact of ST8SiaIV deficiency on the pathophysiological consequences of status epilepticus (SE). ST8SiaIV deficiency reduced the latency to SE induction and increased SE-mediated mortality. Analysis of the doublecortin expression showed a reduced number of neuroblasts as a long-term consequence of SE in ST8SiaIV knockouts. Testing in a battery of different behavioral paradigms indicated that loss of ST8SiaIV affects the long-term behavioral consequences. In summary, the data suggest that the polysialic acid-neural cell adhesion molecule system is a putative target for the modulation of pathophysiological events and affects psychiatric comorbidities in epilepsies.


Expression of Hsp70 in the mud crab, Scylla paramamosain in response to bacterial, osmotic, and thermal stress.
Hsp70 is involved in immune responses against infectious pathogens, thermal, and osmotic stress. To understand the immune defense mechanisms of the mud crab Scylla paramamosain, genomic DNA, transcript level and antimicrobial activities of Hsp70 were analyzed. Genomic DNA sequence analysis revealed one intron in this gene. Furthermore, six SNPs were detected by direct sequencing from 30 samples in this study. Hsp70 mRNA was expressed in almost all tissues examined. By using the quantitative real-time PCR, the expression level of Hsp70 in hemocytes showed a clear time-dependent expression pattern during the 96 h after stimulated by Vibrio alginolyticus. Then, recombinant Hsp70 was obtained by using the bacterial expression system, but no obvious antimicrobial activity has been found for the protein in the antimicrobial tests. After osmotic stress, the expression of Hsp70 in hemocytes showed this gene was induced by the high salinity (30 ‰) for at least 96 h. Hsp70 mRNA expression in hemocytes was analyzed after thermal stress at 6 h, the highest and the lowest expression level of Hsp70 was observed at 36 and 15 °C, respectively. These results indicated that Hsp70 was inducible by bacterial, osmotic, and thermal stress, and therefore plays an important role, different from antibacterial peptide, in innate immune responses of S. paramamosain.


Streptomyces bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).
A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).


Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival.
Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) is a tumor necrosis factor superfamily member that induces apoptosis through the death receptors DR4 and/or DR5 in various cancer cell types but not in most normal cells. Several lung cancer cell lines express DR4 and DR5 and undergo apoptosis in vitro in response to Apo2L/TRAIL. We investigated the efficacy of recombinant soluble human Apo2L/TRAIL and its interaction with chemotherapy in xenograft models based on human NCI-H460 non-small cell lung carcinoma cells. In vitro, Taxol enhanced caspase activation and apoptosis induction by Apo2L/TRAIL. In vivo, Apo2L/TRAIL or Taxol plus carboplatin chemotherapy partially delayed progression of established subcutaneous tumor xenografts, whereas combined treatment caused tumor regression and a substantially longer growth delay. Apo2L/TRAIL, chemotherapy, or the combination of both inhibited growth of preformed orthotopic lung parenchymal tumors versus control by 60%, 57%, or 97%, respectively (all P < 0.01; n = 8-10). Furthermore, combination treatment improved day-90 survival relative to control (7 of 15 versus 1 of 15; P = 0.0003 by Mantel-Cox) as well as to Apo2L/TRAIL (3 of 14; P = 0.031) or chemotherapy (3 of 15; P = 0.035). These studies provide evidence for in vivo activity of Apo2L/TRAIL against lung tumor xenografts and underscore the potential of this ligand for advancing current lung cancer treatment strategies.


Complexed nitrogen heterosuperbenzene: the coordinating properties of a remarkable ligand.
Tetra-peri-(tert-butyl-benzo)-di-peri-(pyrimidino)-coronene 1, the parent compound of the nitrogen heterosuperbenzene family N-HSB, is employed as a novel monotopic ligand in the formation of [Pd(eta3-C3H5)(1)]PF6 2 and [Ru(bpy)2(1)](PF6)2 (where bpy = 2,2'-bipyridine 3a and d8-2,2'-bipyridine 3b). These N-coordinated complexes are fully characterized by 1H NMR and IR spectroscopy and ESI-MS. Metal coordination has a profound effect on both the absorption and the emission properties of 1. Pd(II) coordination causes a red-shift in the low-energy absorptions, a decrease in the intensity of the n-pi absorptions, and a quenching of the emission. Ru(II) coordination causes absorption throughout the visible region and creates two new complexes that join an elite group of compounds known as "black" absorbers. 3a and 3b possess two discernible 1MLCT bands. The one of exceptionally low energy (lambda(max) = 615 nm) has an associated (3)MLCT emission (lambda(max) = 880 nm) due to the unprecedented electron delocalization and acceptor properties of the rigid aromatic N-HSB 1. Both Ru(II) complexes are near-IR emitters with unusually protracted emission lifetimes of 320 ns at 77 K. They are photochemically inert, and their electrochemical properties are consistent with the presence of a low-lying pi orbital on 1. The first two reversible reductions (E(1/2) (CH3CN), -0.54 V, -1.01 V vs SCE) are due to the stepwise reduction of 1 and are anodically shifted as compared to [Ru(bpy)3]2+. Temperature- and concentration-dependent NMR studies on 2 and 3a suggest extensive aggregation is occurring in solution.


Control of retinal ganglion cell axon growth: a new role for Sonic hedgehog.
Retinal ganglion cell (RGC) axons grow towards the diencephalic ventral midline during embryogenesis guided by cues whose nature is largely unknown. We provide in vitro and in vivo evidence for a novel role of Sonic hedgehog (SHH) as a negative regulator of growth cone movement. SHH suppresses both the number and the length of neurites emerging from the chick retina but not from neural tube or dorsal root ganglia explants, without interfering with their rate of proliferation and differentiation. Similarly, retroviral-mediated ectopic expression of Shh along the chick visual pathway greatly interferes the growth of RGC axons. Upon SHH addition to grown neurites, the intracellular level of cAMP decreases, suggesting that the dampening of growth cone extension mediated by SHH may involve interaction with its receptor Patched which is expressed by RGC. Based on these findings, we propose that Shh expression at the chiasm border defines a constrained pathway within the ventral midline which serves to guide the progression of RGC axons.


Mapping the antigenic epitope for a monoclonal antibody against lysozyme.
A monoclonal antibody (HyHEL-5), prepared to chicken lysozyme c by the method of Köhler and Milstein, identified an antigenic site (epitope) that was shared by the lysozymes of seven different species of galliform birds. The lysozymes of two galliform species, bobwhite quail and chachalaca, shared only partial antigenic identity with the epitope defined by this antibody. Duck lysozyme did not react with the antibody at all. Amino acids that determined the epitope structure were tentatively identified by comparing the amino acid sequences of these lysozymes and assuming the antigenic changes produced by evolutionary substitutions are not due to long-range conformational changes. Arg 68 was identified as a determining amino acid. Arg 68 is hydrogen-bonded to Arg 45, and together these two amino acids form a basic cluster that may be a subsite of the epitope. The antibody inhibited lysis of Micrococcus lysodeikticus by chicken lysozyme. Additionally, Biebrich Scarlet, a dye that binds to the catalytic site, inhibited antibody binding to this lysozyme, which indicates that the epitope extends into the cleft region between Arg 45 and Arg 114. The epitope was hypothesized to involve a region measuring at least 13 x 6 x 15 A including the Arg 68-Arg 45 complex that borders the enzymatic catalytic site. Four other monoclonal antibodies to lysozyme have been partially characterized; each had a distinct pattern of binding specificity for various species of bird lysozymes.


Responses to electrical stimulation, noradrenaline, serotonin, and vasopressin in the isolated ear artery of the developing lamb and ewe.
Responses of isolated helical strips of ovine ear artery to electrical stimulation of postganglionic adrenergic neurons and exogenous agonists were studied at various stages of development from 110 days of gestation through to adulthood. Only rudimentary responses were observed at 110-115 days of gestation. A parallel development of responses to noradrenaline (NA), serotonin, and lysine vasopressin began sometime after 110-115 days of gestation and continued until 133-137 days of gestation but there was little development of the latter responses until more than 3-5 days post partum. Development of responses to exogenous agonists was incomplete 2-3 weeks post partum. The development of postganglionic adrenergic responses lagged behind those to exogenous NA. Two to three weeks post partum the NA maximal response was one-third that of adult tissue whereas the response to 16 Hz (highest frequency used) was one-sixth that of adult tissue. The NA threshold concentration was lower in arterial strips of adult animals than it was in those of younger animals. The data suggest that development of functional post-ganglionic adrenergic innervation of vascular smooth muscle begins late in gestation and continues well after birth; this development is preceded by development of vascular mechanisms involved in the response to several agonists.


Assessment of heavy metals in calcium carbide treated mangoes by inductively coupled plasma-mass spectrometry (ICP-MS).
Calcium carbide (CaC2) is widely used as the low price artificial ripening agent of fruits. It may contain traces of heavy metals which can make fruits harmful for human consumption. This study aimed to assess heavy metals present in commercial-grade CaC2 and subsequently their quantification in mango peels for the identification of possible marker elements as an indicator of CaC2 ripening. Samples were prepared by using microwave-assisted digestion followed by multi-elemental quantitative analysis using inductively coupled plasma-mass spectrometry (ICP-MS). In commercial grade CaC2 samples, the concentrations of analysed metals were found to be in the range of 0.6-540.92 µg/g with given order Fe>Zn>Mn>Cr>Pb>Cu>Ag>As>Ni>Co>Cd. The CaC2-treated mango peels showed elevated levels of Fe, Co, As, and Pb in comparison to untreated mango peels (p < 0.05).


Bioactive Peptide Improves Diet-Induced Hepatic Fat Deposition and Hepatocyte Proinflammatory Response in SAMP8 Ageing Mice.
High-fat diet (HFD)-induced nonalcoholic fatty liver disease (NAFLD) poses therapeutic challenges in elderly subjects. Due to lack of efficient drug therapy, plant-based bioactive peptides have been studied as alternative strategy in NAFLD and for less toxicity in elderly. To mimic fatty liver in aging conditions, researchers highly commended the genetically engineered strains SAMP8 (senescence-accelerated mice prone 8). However, there is a paucity of reports about the anti-steatosis effects of bioactive peptides against fatty liver development under a combined action of high-fat diet exposure and aging process. This study was conducted to evaluate the activity of DIKTNKPVIF peptide synthesized from alcalase-generated potato protein hydrolysate (PH), on reducing HFD-driven and steatosis-associated proinflammatory reaction in ageing model. Five groups of six-month-old SAMP8 mice (n=4, each) were fed either a normal chow (NC group) for 14 weeks upon sacrifice, or induced with a 6-week HFD feeding, then treated without (HCO group) or with an 8-week simultaneous administration of peptide (HPEP group), protein (HPH group) or probucol (HRX group). Liver organs were harvested from each group for histological analysis and immunoblot assay. In contrast to NC, extensive fat accumulation was visualized in the liver slides of HCO. Following the trends of orally administered PH, intraperitoneally injected peptide reduces hepatic fat deposition and causes at protein level, a significant decrease in HFD-induced proinflammatory mediators p-p38 MAPK, FGF-2, TNF-α, IL-6 with concomitant reactivation of AMPK. However, p-Foxo1 and PPAR-α levels were slightly changed. Oral supplementation of PH and intraperitoneal injection of derived bioactive peptide alleviate proinflammatory reaction associated with hepatosteatosis development in elderly subjects, through activation of AMPK.


Structure of laboratory ball lightning.
Trajectories of self-sustained laboratory ball lightning, generated by arc discharges with silicon, are investigated for understanding the possibility of buoyant flight. Extremely low apparent densities are found, nearly approaching that of standard air. The freely buoyant balls are observed to survive for about 0.1 s, with significantly buoyant balls surviving for several seconds. These ball lightning objects are found to have a density and size that can easily allow them to be carried by a gentle breeze of a few meters per second. The results are interpreted by a model that is an extension of that first proposed by Abrahamson and Dinniss [J. Abrahamson and J. Dinniss, Nature (London) 403, 519 (2000)]. The buoyant behavior of ball lightning seen in our experiments is believed to arise as a result of the formation of a nanoparticle oxide network growing from a molten silicon core.


Activation of the primary motor cortex using fully-implanted electrical sciatic nerve stimulation.
Functional degradation of the motor cortex usually results from brain injury, stroke, limb amputation, aging or other diseases. Currently, there are no ideal means of treatment, other than medication and sports rehabilitation. The present study investigated whether electrical stimulation of the sciatic nerve can activate the motor-related area of the brain. The study is based on a self-developed fully implantable nerve electrical stimulator and a self-developed multi-channel electroencephalogram (EEG) electrode array. The sciatic nerves of Sprague-Dawley rats (sorted into old and young groups) were stimulated by the electrical stimulator under anesthesia, and the EEG signal was recorded simultaneously. The relationship between sciatic nerve stimulation and brain activity was analyzed. The results showed that when the sciatic nerve was stimulated by the implanted electrical stimulator, motor-related channels were activated, causing contraction of the left leg. It was found that at the frequency band of 8-16 Hz, the EEG signal in the right motor area was higher than at other frequency bands. This phenomenon was identical in both young and old rats. The results indicated that electrical stimulation of the sciatic nerve can activate the motor region of the rat brain, and provided evidence that stimulation of the sciatic nerve could be a method of preventing motor cortex degeneration.


Multivalence Charge Transfer in Doped and Codoped Photocatalytic TiO2.
The present work reports data for the mineralogical and chemical properties of anatase thin films individually doped or codoped with chromium and vanadium, fabricated by sol-gel spin coating on glass substrates and annealing at 450 °C for 2 h. X-ray photoelectron spectroscopy data indicated the presence of Ti(4+), Ti(3+), Cr(3+), and possibly Cr(4+) in the Cr-doped thin films; Ti(4+), Ti(3+), V(3+), V(4+), and possibly V(5+) in the V-doped thin films; and Ti(4+), Ti(3+), Cr(3+), Cr(4+), V(3+), V(4+), and possibly V(5+) in the codoped thin films. While the thermodynamically stable valences Ti(4+), Cr(3+), and V(5+) would be expected to have formed, the presence of the nonequilibrium valences Ti(3+), Cr(4+), V(3+), and V(4+) is considered to have resulted from intervalence charge transfer for the Cr-doped and V-doped systems but from multivalence charge transfer (MVCT) for the codoped system. The latter phenomenon, which is introduced as a new conceptual term, describes the nature of the mutual exchange of electrons during valence changes of both dopant (Cr, V) and matrix (Ti) ions during annealing. In the present case, MVCT appears to be a transient metastable condition that acts during annealing, but subsequent UV irradiation can alter its effects.


Familial Aggregation of Psoriasis and Co-Aggregation of Autoimmune Diseases in Affected Families.
Psoriasis is considered to result from the interaction of genetic factors and environmental exposure. The evidence for familial aggregation in psoriasis has been reported but population-based studies related to the magnitude of genetic contribution to psoriasis are rare. This study aimed to evaluate the relative risks of psoriasis in individuals with affected relatives and to calculate the proportion of genetic, shared, and non-shared environmental factors contributing to psoriasis. The study cohort included 69,828 patients diagnosed with psoriasis enrolled in National health Insurance in 2010. The adjusted relative risks (RR) for individuals with an affected first-degree relative and affected second-degree relative were 5.50 (95% CI (Confidence Interval), 5.19⁻5.82) and 2.54 (95% CI, 2.08⁻3.12) respectively. For those who have affected first-degree relatives, their RR was 1.45 (95% CI, 1.17⁻1.79) for Sjogren's syndrome and 1.94 (95% CI, 1.15⁻3.27) for systemic sclerosis. This nationwide study ascertains that family history of psoriasis is a risk factor for psoriasis. Individuals with relatives affected by psoriasis have higher risks of developing some autoimmune diseases.


Schwann cells promote the capability of neural stem cells to differentiate into neurons and secret neurotrophic factors.
The present study investigated whether co-culturing Schwann cells (SCs) with neural stem cells (NSCs) improves viability, direction of differentiation and secretion of brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF) in NSCs. The three groups assessed were as follows: SCs, NSCs, and a co-culture of SCs and NSCs. Cellular morphological changes were observed under an inverted phase contrast microscope and quantified. Cells were identified by immunofluorescence staining: S100 for SCs, Nestin for NSCs, microtubule associated protein (Map) 2 and NeuN for neurons and glial fibrillary acidic protein for astrocytes. Cell viability was evaluated by MTT assay. Secretion of BDNF and GDNF was quantified; mRNA expression was quantified by reverse transcription-quantitative polymerase chain reaction. The majority of NSCs in the co-cultured group differentiated into neurons. The cell survival rate of the co-culture group was significantly higher than the other groups on days 3, 5 and 10 (P<0.01). The secretion of BDNF in the co-culture group was significantly higher than NSCs on days 3, 5 and 7 (P<0.05), while the amount of GDNF in co-culture was significantly higher than both NSCs and SCs on day 1 (P<0.05). BDNF and GDNF gene expression in the co-culture group was significantly higher than SCs (P<0.01). Gene expression of Map2 in co-culture group was also significantly higher than both NSC and SC groups (P<0.01). Therefore, co-cultured SCs and NSCs promote differentiation of NSCs into neurons and secrete higher levels of neurotropic factors including BDNF and GDNF.


Do antihypertensive drugs precipitate diabetes?
A longitudinal population study of 1462 women aged 38-60 was carried out from 1968-9 to 1980-1 in Gothenburg, Sweden. The initial and follow up examinations included questions concerning history of diabetes and antihypertensive treatment. A considerably increased risk of developing diabetes was observed for subjects with hypertension taking diuretics (895 patient years), subjects taking beta blockers (682 patient years), and subjects taking a combination of diuretics and beta blockers (281 patient years) compared with subjects not taking antihypertensive drugs (13 855 control years). When diuretics and beta blockers were compared no difference was found in relative risk. Despite this increased risk, and because little is known about the relation between other forms of antihypertensive treatment and diabetes, diuretics and beta blockers should remain the treatments of choice in arterial hypertension.


Effects of Phlebotomy on Liver Enzymes and Histology of Patients with Nonalcoholic Fatty Liver Disease.
Nonalcoholic fatty liver disease (NAFLD), defined as excessive liver fat deposition and one of end-stage liver disease causes. Increased ferritin levels are associated with insulin resistance and a higher hepatic iron and fat content. Hyperferritinemia has been associated with severity of liver damage in NAFLD. The study aimed to evaluate the effects of phlebotomy on liver enzymes and histology in such patients. Thirty-two eligible patients who had NAFLD and after 6 months of lifestyle modification still had NAFLD, and whose ferritin serum was above 250 mg/dl, were enrolled in this clinical trial study. After written informed consent was obtained, each patient's blood serum was taken for aspartate transaminase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALK-P), complete blood count (CBC), total iron-binding capacity (TIBC), iron, and ferritin. Then the patients underwent liver biopsy. After that patients underwent phlebotomy, giving 350 cc blood monthly. Before every phlebotomy, hemoglobin and ferritin were checked. If they were in the goal range, phlebotomy was discontinued and the patient underwent liver biopsy. A serum sample was taken for testing at the beginning of the study. The results before and after phlebotomy were compared. The maximum duration of the study was 6 months. Thirty-two patients (26 males and 6 females) were enrolled, and the mean average age was 33.7 ± 6.74 years. Phlebotomy improved liver enzymes and histology of liver significantly (P < 0.001) and induced reduction of ferritin. Phlebotomy is effective for the improvement of liver enzymes and histology in patients with NAFLD and hyperferritinemia.


Effective inhaled drug administration to mechanically ventilated patients.
Inhaled therapy is commonly employed in mechanically ventilated patients with chronic obstructive pulmonary disease or asthma. The efficacy of inhaled drugs is comparable to that achieved with systemic routes of administration, but the dose of drug required to achieve a therapeutic effect is generally much smaller. Moreover, limited systemic absorption of inhaled drugs minimises systemic side effects. Aerosol administration to ventilated patients differs from that in ambulatory patients in several respects. Optimal techniques for using pressurised metered-dose inhalers and nebulisers in ventilator circuits have been developed. With these techniques, the efficiency of inhaled drug delivery in mechanically ventilated patients is now comparable to that in ambulatory patients. Pressurised metered-dose inhalers are chiefly used to deliver bronchodilator and corticosteroid aerosols, and are more efficient and convenient to use than nebulisers for routine therapy in ventilated patients. However, nebulisers are more versatile and are employed to generate aerosols of bronchodilators, corticosteroids, antibiotics, prostaglandins, surfactant and mucolytic agents. Improvements in drug formulations and the design and efficiency of aerosol generating devices have led to increasing application of inhaled therapies in mechanically ventilated patients.


Hemispheric asymmetry, early visual processes, and serial memory comparison.
Observers indicated whether a single probe letter presented to the left visual field/right hemisphere (LVF-RH) or to the right visual field/left hemisphere (RVF-LH) matched one of two, three, four, or five set letters in both name and case. For positive trials during the initial experimental session, the slope of the linear memory set size reaction time function was increased by perceptually degrading the probe letter on LVF-RH trials, but not on RVF-LH trials. In addition, perceptual degradation of the probe letter increased the intercept of the memory set size function more on RVF-LH trials than on LVF-RH trials. During subsequent experimental sessions, the same pattern of intercept results was obtained but perceptual degradation of the probe no longer changed the slope for either LVF-RH or RVF-LH trials. However, the slopes were uniformly lower on RVF-LH trials than on LVF-RH trials. The major results are consistent with hypothesized right-hemisphere efficiency for early processing stages and left-hemisphere efficiency for serial processing of alphanumeric information. The results further illustrate the importance of separating stages of processing in studies of cerebral laterality and indicate that the relative difficulty of the various stages can be a determinant of laterality results.


Worldwide co-occurrence analysis of 17 species of the genus Brachypodium using data mining.
The co-occurrence of plant species is a fundamental aspect of plant ecology that contributes to understanding ecological processes, including the establishment of ecological communities and its applications in biological conservation. A priori algorithms can be used to measure the co-occurrence of species in a spatial distribution given by coordinates. We used 17 species of the genus Brachypodium, downloaded from the Global Biodiversity Information Facility data repository or obtained from bibliographical sources, to test an algorithm with the spatial points process technique used by Silva et al. (2016), generating association rules for co-occurrence analysis. Brachypodium spp. has emerged as an effective model for monocot species, growing in different environments, latitudes, and elevations; thereby, representing a wide range of biotic and abiotic conditions that may be associated with adaptive natural genetic variation. We created seven datasets of two, three, four, six, seven, 15, and 17 species in order to test the algorithm with four different distances (1, 5, 10, and 20 km). Several measurements (support, confidence, lift, Chi-square, and p-value) were used to evaluate the quality of the results generated by the algorithm. No negative association rules were created in the datasets, while 95 positive co-occurrences rules were found for datasets with six, seven, 15, and 17 species. Using 20 km in the dataset with 17 species, we found 16 positive co-occurrences involving five species, suggesting that these species are coexisting. These findings are corroborated by the results obtained in the dataset with 15 species, where two species with broad range distributions present in the previous dataset are eliminated, obtaining seven positive co-occurrences. We found that B. sylvaticum has co-occurrence relations with several species, such as B. pinnatum, B. rupestre, B. retusum, and B. phoenicoides, due to its wide distribution in Europe, Asia, and north of Africa. We demonstrate the utility of the algorithm implemented for the analysis of co-occurrence of 17 species of the genus Brachypodium, agreeing with distributions existing in nature. Data mining has been applied in the field of biological sciences, where a great amount of complex and noisy data of unseen proportion has been generated in recent years. Particularly, ecological data analysis represents an opportunity to explore and comprehend biological systems with data mining and bioinformatics tools.


Rationale and design for the PAINT randomized trial.
We describe the rationale and design for the 'PercutAneous INTervention with biodegradable-polymer based paclitaxel-eluting or sirolimus-eluting versus bare stents for de novo coronary lesions - PAINT trial'. To evaluate two novel formulations of paclitaxel-eluting stent and the sirolimus-eluting stent against a stent with the same metallic structure but without polymer coating or drug elution. The PAINT is a multicenter 3-arm randomized trial, conducted in Brazilian tertiary institutions, which included 275 patients allocated for the InfinniumR paclitaxel-eluting stent, the SupralimusR sirolimus-eluting stent or the Milennium MatrixR bare metal stent in a 2:2:1 ratio. Patients had de novo coronary lesions in native vessels with a diameter between 2.5 and 3.5 mm, amenable for treatment with a single stent of 29 mm or less in length. The primary objective was to compare the in-stent late loss at 9 months of both paclitaxel- and sirolimus-eluting versus the late loss of control bare metal stents. Important secondary objectives included the comparison in outcomes between sirolimus and paclitaxel stents, as well as the analysis of the incidence of major adverse cardiac events. The PAINT trial had a unique design that allowed for the evaluation of the safety and efficacy profiles of two novel drug-eluting stent formulations, with a biodegradable-polymer carrier and releasing paclitaxel or sirolimus, which were compared against a bare metal stent (primary objective). As the drug-eluting stents differed by the drug, but were identical otherwise, the trial also allowed the comparison of the anti-restenosis effects of sirolimus versus paclitaxel (secondary objective).


Using field data analysis for environmental decision making and subsequent remediation at two example sites.
One of the major challenges in remediating contaminated sites is having quick access to quality data on which to base remedial decisions as onsite work progresses. Case studies are presented at two Superfund sites where field screening and field analyses are used to provide these data. Emphasis is placed on the importance of high quality field data, as these data are the basis for remedial decisions prior to receipt of offsite laboratory confirmation. The decision-making processes for remediating contaminated soils and structures are presented in addition to project specifics including data quality objectives, field data collection procedures, quality assurance/quality control procedures, and comparisons of the field data with offsite laboratory results.


Ultrastructural confirmation of malignant melanoma.
We have recently had the opportunity to examine two cases, where malignant melanoma was suspected on light microscopy and Fontana-Masson stain was focally and faintly positive. Electron microscopy of four to five different blocks failed, however, to demonstrate the presence of premelanosomes and thereby to confirm the diagnosis of malignant melanoma. The areas in the paraffin blocks that on Fontana-Masson stain had showed a positive reaction were deparaffinized, embedded in plastic, and examined by electron microscopy. Now the characteristic premelanosomes were found and the diagnosis of malignant melanoma was confirmed.


Performance of two blood culture systems to detect anaerobic bacteria. Is there any difference?
We studied the performance characteristics of two blood culture (BC) bottles/systems, (i) BacT/ALERT-FN Plus/3D (bioMérieux, Marcy l'Étoile, France) and (ii) BACTEC-Lytic/9000 (Becton Dickinson, Sparks, USA) for detection of growth and time-to-positivity (TTP) against a balanced and diverse collection of anaerobic bacterial strains (n = 48) that included reference strains (n = 19) and clinical isolates (n = 29) of 32 species (15 Gram-negative and 17 Gram-positive). Standard suspension of bacteria was inoculated to each bottle in duplicates and incubated in the corresponding system. Overall, 62.5% (n = 30) of strains were detected by both BC bottle types. Comparing the two, 70.8% (n = 34) and 79.2% (n = 38) of strains were detected by BacT/ALERT-FN Plus and BACTEC-Lytic bottles, respectively (p = 0.38). Among Gram-negative anaerobes (n = 25) the detection rate was 76.0% (n = 19) vs. 92.0% (n = 23) (p = 0.22), respectively. Among Gram-positive anaerobes (n = 23) the detection rate was 65.2% (n = 15) in both bottles (p = 1). The average TTP per bottle was calculated only for the strains detected by both systems (n = 30) and was 40.85 h and 28.08 h for BacT/ALERT-FN Plus and BACTEC-Lytic, respectively (p < 0.001). The mean difference was 12.76 h (95% CI: 6.21-19-31 h). Six anaerobic strains were not detected by any system, including Gram-negative Porphyromonas gingivalis, and five Gram-positive strains: Finegoldia magna, Peptostreptococcus anaerobius, Propionibacterium acnes, Clostridium novyi and Clostridium clostridioforme. Furthermore, Eggerthella lenta and Prevotella bivia were detected only by BacT/ALERT-FN Plus, while Prevotella disiens and Prevotella intermedia were detected only by BACTEC-Lytic bottles. There were no major differences in detection rate among clinical and reference strains. Anaerobic bacteria represent a minority of BC isolates, however, far from ideal detection rate was observed in this study for both tested bottle/system combinations. Nevertheless, in those cases where both gave positive signal, BACTEC-Lytic was superior to BacT/ALERT FN Plus with 12.76 h shorter mean TTP. Improvements of media in blood culture bottles available for detection of anaerobes are warranted.


Induction of secondary immune response by reactivated Japanese encephalitis virus in latently infected mice.
Development of secondary immune response has been studied following reactivation of latent Japanese encephalitis virus (JEV) infection in mice. The virus could be reactivated in 43% of the latently infected mice at 27 weeks p.i. by treatment with cyclophosphamide. The reactivated virus induced delayed-type hypersensitivity (DTH) and leucocyte migration inhibition (LMI) responses in mice, with peak activity on Day 5 post-reactivation (p.r.). The DTH persisted at low levels for long periods. Humoral immunity measured by haemagglutination-inhibiting antibody showed a four-fold rise in antibody titres. DTH was transferable by immune spleen cells for 5 days p.r. only. It is, therefore, concluded that JEV reactivation generates a quick and short-lived secondary immune response.


MicroRNA-23a-5p Is Involved in the Regulation of Lipopolysaccharide-Induced Acute Lung Injury by Targeting HSP20/ASK1.
Inflammation and oxidative stress contribute to the progression of acute lung injury (ALI). MicroRNA-23a-5p (miR-23a-5p) has been reported to regulate inflammation and oxidative stress; however, its role in ALI is still poorly elucidated. Mice were intravenously treated with the miR-23a-5p antagomir, agomir, or the negative controls for 3 consecutive days and then received a single intratracheal injection of lipopolysaccharide (LPS, 5 mg/kg) to induce ALI. Pulmonary function, bronchoalveolar lavage fluids (BALFs), arterial blood gas, and molecular biomarkers associated with inflammation and oxidative stress were analyzed. In addition, murine peritoneal macrophages were isolated and treated with LPS to verify the role of miR-23a-5p in vitro. We detected an elevation of miR-23a-5p expression in the lungs from ALI mice. The miR-23a-5p antagomir was prevented, whereas the miR-23a-5p agomir aggravated inflammation, oxidative stress, lung tissue injury, and pulmonary dysfunction in LPS-treated mice. Besides, the miR-23a-5p antagomir also reduced the productions of proinflammatory cytokines and free radicals in LPS-treated primary macrophages, which were further augmented in cells following the miR-23a-5p agomir treatment. Additional findings demonstrated that the miR-23a-5p agomir exacerbated LPS-induced ALI via activating apoptosis signal-regulating kinase 1 (ASK1), and that pharmacological or genetic inhibition of ASK1 significantly repressed the deleterious effects of the miR-23a-5p agomir. Moreover, we proved that the miR-23a-5p agomir activated ASK1 via directly reducing heat shock protein 20 (HSP20) expression. miR-23a-5p is involved in the regulation of LPS-induced inflammation, oxidative stress, lung tissue injury, and pulmonary dysfunction by targeting HSP20/ASK1, and it is a valuable therapeutic candidate for the treatment of ALI.


Acute Otitis due to Vibrio fluvialis after Swimming.
A 40-year-old female presented with purulent exudate through the left auditive duct and pain in the left ear region, which intensified during mastication. After collection of the pus from the left ear lesion, amoxicillin-clavulanic acid for seven days was prescribed for a presumed diagnosis of acute otitis. Four days later, the pus culture grew V. fluvialis which is further identified by API 20E identification system (bioMérieux). Following the successful completion of a course of antibiotics, the patient recovered completely and without complication. To the best of our knowledge, this is the first case of Vibrio fluvialis otitis after swimming in an immunocompetent patient.


Regioselective biolistic targeting in organotypic brain slices using a modified gene gun.
Transfection of DNA has been invaluable for biological sciences and with recent advances to organotypic brain slice preparations, the effect of various heterologous genes could thus be investigated easily while maintaining many aspects of in vivo biology. There has been increasing interest to transfect terminally differentiated neurons for which conventional transfection methods have been fraught with difficulties such as low yields and significant losses in viability. Biolistic transfection can circumvent many of these difficulties yet only recently has this technique been modified so that it is amenable for use in mammalian tissues. New modifications to the accelerator chamber have enhanced the gene gun's firing accuracy and increased its depths of penetration while also allowing the use of lower gas pressure (50 psi) without loss of transfection efficiency as well as permitting a focused regioselective spread of the particles to within 3 mm. In addition, this technique is straight forward and faster to perform than tedious microinjections. Both transient and stable expression are possible with nanoparticle bombardment where episomal expression can be detected within 24 hr and the cell survival was shown to be better than, or at least equal to, conventional methods. This technique has however one crucial advantage: it permits the transfection to be localized within a single restrained radius thus enabling the user to anatomically isolate the heterologous gene's effects. Here we present an in-depth protocol to prepare viable adult organotypic slices and submit them to regioselective transfection using an improved gene gun.


Identifying Novel Types of Irritability Using a Developmental Genetic Approach.
Irritability, which is strongly associated with impairment and negative outcomes, is a common reason for referral to mental health services but is a nosological and treatment challenge. A major issue is how irritability should be conceptualized. The authors used a developmental approach to test the hypothesis that there are several forms of irritability, including a "neurodevelopmental/ADHD-like" type, with onset in childhood, and a "depression/mood" type, with onset in adolescence. Data were analyzed from the Avon Longitudinal Study of Parents and Children, a prospective U.K. population-based cohort. Irritability trajectory classes were estimated for 7,924 individuals with data at multiple time points across childhood and adolescence (four possible time points from approximately ages 7 to 15). Psychiatric diagnoses were assessed at approximately ages 7 and 15. Psychiatric genetic risk was indexed by polygenic risk scores (PRSs) for attention deficit hyperactivity disorder (ADHD) and depression, derived using large genome-wide association study results. Five irritability trajectory classes were identified: low (81.2%), decreasing (5.6%), increasing (5.5%), late-childhood limited (5.2%), and high-persistent (2.4%). The early-onset high-persistent trajectory was associated with male preponderance, childhood ADHD (odds ratio=108.64, 95% CI=57.45-204.41), and ADHD PRS (odds ratio=1.31, 95% CI=1.09-1.58). The adolescent-onset increasing trajectory was associated with female preponderance, adolescent depression (odds ratio=5.14, 95% CI=2.47-10.73), and depression PRS (odds ratio=1.20, 95% CI=1.05-1.38). Both the early-onset high-persistent and adolescent-onset increasing trajectory classes were associated with adolescent depression diagnosis and ADHD PRS. The developmental context of irritability may be important in its conceptualization: early-onset persistent irritability may be more neurodevelopmental/ADHD-like and later-onset irritability more depression/mood-like. These findings have implications for treatment as well as nosology.


Conservation of noncoding microsatellites in plants: implication for gene regulation.
Microsatellites are extremely common in plant genomes, and in particular, they are significantly enriched in the 5' noncoding regions. Although some 5' noncoding microsatellites involved in gene regulation have been described, the general properties of microsatellites as regulatory elements are still unknown. To address the question of microsatellites associated with regulatory elements, we have analyzed the conserved noncoding microsatellite sequences (CNMSs) in the 5' noncoding regions by inter- and intragenomic phylogenetic footprinting in the Arabidopsis and Brassica genomes. We identified 247 Arabidopsis-Brassica orthologous and 122 Arabidopsis paralogous CNMSs, representing 491 CT/GA and CTT/GAA repeats, which accounted for 10.6% of these types located in the 500-bp regions upstream of coding sequences in the Arabidopsis genome. Among these identified CNMSs, 18 microsatellites show high conservation in the regulatory regions of both orthologous and paralogous genes, and some of them also appear in the corresponding positions of more distant homologs in Arabidopsis, as well as in other plants. A computational scan of CNMSs for known cis-regulatory elements showed that light responsive elements were clustered in the region of CT/GA repeats, as well as salicylic acid responsive elements in the (CTT)n/(GAA)n sequences. Patterns of gene expression revealed that 70-80% of CNMS (CTT)n/(GAA)n associated genes were regulated by salicylic acid, which was consistent with the prediction of regulatory elements in silico. Our analyses showed that some noncoding microsatellites were conserved in plants and appeared to be ancient. These CNMSs served as regulatory elements involved in light and salicylic acid responses. Our findings might have implications in the common features of the over-represented microsatellites for gene regulation in plant-specific pathways.


The permanent dipole moment of gas-phase para-amino benzoic acid revisited.
The permanent dipole moment of para-amino benzoic acid has been calculated at various theoretical levels, including Hartree-Fock, second-order Møller-Plesset perturbation (MP2), coupled cluster singles and doubles (CCSD), and triples corrections CCSD(T), and hybrid density functional theory at B3LYP level. It is found that the B3LYP method fails to provide correct results for the geometry and the permanent dipole moment. These results are significantly improved by MP2 calculations. Our best estimated dipole moments obtained at CCSD and CCSD(T) levels are in good agreement with experiment.


Engineering antiphagocytic biomimetic drug carriers.
Drug-delivery carriers have the potential to not only treat but also diagnose many diseases; however, they still lack the complexity of natural-particulate systems. Cell-based therapies using tumor-targeting T cells and tumor-homing mesenchymal stem cells have given researchers a means to exploit the characteristics exhibited by innate-biological entities. Similarly, immune evasion by pathogens has inspired the development of natural polymers to cloak drug carriers. The 'marker-of-self' CD47 protein, which is found ubiquitously on mammalian cell surfaces, has been used for evading phagocyte clearance of drug carriers. This review will focus on the recent progress of drug carriers co-opting the tricks that cells in nature use to hide safely under the radar of the body's innate immune system.


Vibrational dynamics of DNA. III. Molecular dynamics simulations of DNA in water and theoretical calculations of the two-dimensional vibrational spectra.
A theoretical description of the vibrational excitons in DNA is presented by using the vibrational basis mode theory developed in Papers I and II. The parameters obtained from the density functional theory calculations, such as vibrational coupling constants and basis mode frequencies, are used to numerically simulate two-dimensional (2D) IR spectra of dG(n):dC(n) and dA(n):dT(n) double helices with n varying from 1 to 10. From the molecular dynamics simulations of dG(5)C(5) and dA(5)T(5) double helices in D(2)O solution, it is found that the thermally driven internal motions of these systems in an aqueous solution do not induce strong fluctuations of basis mode frequencies nor vibrational couplings. In order to construct the two-exciton Hamiltonian, the vibrational anharmonicities of eight basis modes are obtained by carrying out B3LYP6-31G(*) calculations for the nine basis modes. The simulated 2D IR spectra of dG(n):dC(n) double helix in D(2)O solution are directly compared with closely related experimental results. The 2D IR spectra of dG(n):dC(n) and dA(n):dT(n) are found to be weakly dependent on the number of base pairs. The present work demonstrates that the computational procedure combining quantum chemistry calculation and molecular dynamics simulation methods can be of use to predict 2D IR spectra of nucleic acids in solutions.


Using biological inspiration to engineer functional nanostructured materials.
Humans have always looked to nature for design inspiration, and material design on the molecular level is no different. Here we explore how this idea applies to nanoscale biomimicry, specifically examining both recent advances and our own work on engineering lipid and polymer membrane systems with cellular processes.


Strategies for long-term preservation of kidney graft function.
Kidney transplantation has become a routine procedure in the treatment of patients with kidney failure, and requires collaboration of experts from different disciplines, such as nephrology, surgery, immunology, pathology, infectious disease medicine, cardiology, and oncology. Grafts can be obtained from deceased or living donors, with different logistical requirements and implications for long-term graft patency. 1-year graft survival rates are greater than 95% in many centres but improvement of long-term function remains a challenge. New developments in molecular immunology and computational biology have increased precision of donor and recipient matching of HLA and non-HLA compatibility. Individual omics-wide molecular diagnostics, extracorporeal therapies, and drug developments allow for precise individual decision making and treatment. Tolerance induction by mixed chimerism without toxic conditioning and with a low risk of graft versus host disease is a visionary but realistic goal. Some of these innovations are already used in modern transplant centres and will allow advancement in long-term allograft preservation.


Evaluation of LAMP Assay Using Phenotypic Tests and Conventional PCR for Detection of nuc and mecA genes Among Clinical Isolates of Staphylococcus spp.
The purpose of this study is to develop a nuc and mecA gene specific Loop-mediated isothermal Amplification (LAMP) assay for rapid identification and detection of methicillin resistant Staphylococcus aureus among clinical isolates. A total of 100 (70 from pus and 30 from blood), clinical isolates of Staphylococcus spp were screened for the nuc gene to differentiate between S.aureus and Coagulase negative Staphylococci (CONS) by a nuc gene specific LAMP assay. The isolates were also screened for the presence of the mec Agene by the mecA specific LAMP assay. The results were compared with the phenotypic identification and methicillin resistance by Vitek-2 system (bioMérieux, Marcy l'Etoile, France) and conventional PCR. Among 100 Staphylococcus isolates, there were 82 (82%) Staphylococcus aureus isolates and 18 (18%) coagulase negative Staphylococcus as detected by the Vitek 2, conventional PCR and the LAMP assay using the nuc gene. The mecA gene was detected by the LAMP assay in 56(56%) isolates (44 Methicillin resistant Staphylococcus aureus (MRSA) and 12 Methicillin resistant coagulase negative Staphylococcus (MRCONS), which were also identified by the Vitek 2 and conventional PCR as methicillin resistant. The results of the LAMP assay were available within 90min as compared to the Vitek 2 results (18- 24hours) and conventional PCR (3-4 hours). The present study proved that LAMP assay can be used for the simultaneous differentiation of Staphylococcal spp and detection of methicillin resistance.


Terra firma-forme dermatosis revisited.
Terra firma-forme dermatosis is a cutaneous discoloration resembling dirt, hence the clinical name. The dyschromia cannot be removed with routine soap and water washing, but can be eliminated by rubbing with isopropyl alcohol. Although the condition poses no serious medical threat, it is cosmetically distressing. Our experience is that this entity is much more frequent that might be expected when considering the paucity of reports in the readily available medical literature. We chronicle a series of cases to highlight occurrence in widely divergent demographic groups and long duration of remission following appropriate topical intervention. The cause remains unknown.


Use of umbilical cord blood for stem cell research.
Umbilical cord blood (UCB), long treated as waste material, is today considered a valuable source of hematopoietic stem cells. UCB is used, mostly in children, for the treatment of blood malignancies and inherited blood and metabolic disorders. In addition to blood precursor cells, UCB also contains stem cells that can differentiate into other types, such as cartilage, fat, hepatic, cardiac, and neural cells, fuelling speculation about the use of cord blood stem cells for regenerative medicine. Further research is therefore needed to investigate the expanded potential of UCB and its therapeutic use in cell and tissue therapies. According to a recent survey, practices for the procurement of UCB for research vary widely across Canada, so this area may not yet be ready for uniform regulation. However, some harmonization of practices to increase the availability of UCB for research would be useful for Canadian investigators. In this article, we address several important ethical and legal issues relating to the use of UCB in research and recommend guidelines to serve as a source of useful information for researchers. While their legal acceptability may vary across Canada, it is hoped that these recommendations foster more harmonized UCB research practices.


[Enzymatic synthesis of high-modified DNA].
Here, we describe the synthesis and purification of six deoxyuridine triphosphate derivatives that contain protein-like functional groups and alkene linkers of various lengths. Using KOD XL and Deep Vent polymerases, these derivatives have been incorporated into single-stranded DNA, achieving a high degree of DNA modification. These polymerases are able to utilize highly modified DNA strands as templates for synthesizing unmodified DNA. The synthesized deoxyuridine triphosphate derivatives are promising as substrates for producing modified aptamers to various target proteins using, e.g., the systematic evolution of ligands by exponential enrichment (SELEX) methodology.


Molecular characterization of Lactobacillus plantarum DMDL 9010, a strain with efficient nitrite degradation capacity.
Nitrites commonly found in food, especially in fermented vegetables, are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. A Lactobacillus strain (Lactobacillus sp. DMDL 9010) was previously isolated from fermented vegetables by our group, and is not yet fully characterized. A number of phenotypical and genotypical approaches were employed to characterize Lactobacillus sp. DMDL 9010. Its nitrite degradation capacity was compared with four other Lactobacillus strains, including Lactobacillus casei subsp. rhamnosus 719, Lactobacillus delbrueckii subsp. bulgaricu 1.83, Streptococcus thermophilus 1.204, and lactobacillus plantarum 8140, on MRS medium. Compared to these four Lactobacillus strains, Lactobacillus sp. DMDL 9010 had a significantly higher nitrite degradation capacity (P<0.001). Based on 16S rDNA sequencing and sequence comparison, Lactobacillus sp. DMDL 9010 was identified as either Lactobacillus plantarum or Lactobacillus pentosus. To further identify this strain, the flanking regions (922 bp and 806 bp upstream and downstream, respectively) of the L-lactate dehydrogenase 1 (L-ldh1) gene were amplified and sequenced. Lactobacillus sp. DMDL 9010 had 98.92 and 76.98% sequence identity in the upstream region with L. plantarum WCFS1 and L. pentosus IG1, respectively, suggesting that Lactobacillu sp. DMDL 9010 is an L. plantarum strain. It was therefore named L. plantarum DMDL 9010. Our study provides a platform for genetic engineering of L. plantarum DMDL 9010, in order to further improve its nitrite degradation capacity.


The carboxyl terminus of RNA helicase A contains a bidirectional nuclear transport domain.
Human RNA helicase A was recently identified to be a shuttle protein which interacts with the constitutive transport element (CTE) of type D retroviruses. Here we show that a domain of 110 amino acids at the carboxyl terminus of helicase A is both necessary and sufficient for nuclear localization as well as rapid nuclear export of glutathione S-transferase fusion proteins. The import and export activities of this domain overlap but are separable by point mutations. This bidirectional nuclear transport domain (NTD) has no obvious sequence homology to previously identified nuclear import or export signals. However, the Ran-dependent nuclear import of NTD was efficiently competed by excess amounts of the nuclear localization signal (NLS) peptide from simian virus 40 large T antigen, suggesting that import is mediated by the classical NLS pathway. The nuclear export pathway accessed by NTD is insensitive to leptomycin B and thus is distinct from the leucine-rich nuclear export signal pathway mediated by CRM1.


The Jolly-Seber model with tag loss.
Tag loss in mark-recapture experiments is a violation of one of the Jolly-Seber model assumptions. It causes bias in parameter estimates and has only been dealt with in an ad hoc manner. We develop methodology to estimate tag retention and abundance in double-tagging mark-recapture experiments. We apply this methodology to walleyes (Stizostedion vitreum) in Mille Lacs, Minnesota.


Identifying emotions in music through electrical hearing in deaf children using cochlear implants.
Cochlear implant (CI) devices are the choice of treatment for individuals with severe to profound hearing loss. The CI devices provide the opportunity for children who are deaf to perceive sound by electrical stimulation of the auditory nerve, with the goal of optimizing oral communication. A natural benefit of acquiring hearing using CIs is the ability to hear, and perhaps enjoy, music. Music is a non-verbal auditory stimulus and a powerful tool for transmitting emotion. Identifying emotional cues is an important part of normal social development and communication and thus music may play an important role in establishing these skills during development. To date, it is not known whether children who use cochlear implants to hear can identify the emotional content carried in music. Our objective in the present study was to determine whether children who have been deaf from infancy and are experienced CI users have acquired the ability to identify emotion in musical phrases. Study participants were 18 CI users (ages 7-13 years) who received right unilateral CIs (mean age at CI activation of 2.9 years) and 18 age-and gender-matched controls. Participants were asked to judge 32 brief musical excerpts as happy or sad by pointing to simple graphics of a smiling or frowning face. Children using CIs were able to correctly distinguish happy versus sad music well above chance levels, but performed more poorly on this task than their peers with typical hearing. Age at CI activation and time since CI activation were both uncorrelated with outcome measures. Children with CIs show the ability to perceive emotion in music but do so less accurately than typically hearing peers.


Performance of a low-frequency, multi-resonant broadband Tonpilz transducer.
The underwater performance of a high-power multi-resonant Tonpilz transducer with a nearly flat frequency response and a power handling capability of 2 kW (peak) is reported here. A maximum transmitting voltage response (TVR) value of 156 dB re: 1 microPa/V at 1 m has been achieved at 3 kHz with a specially designed matching coil. A maximum receiving sensitivity (RS) of -164 dB re: 1 V/microPa at 3 kHz has been measured without using a matching coil. The horizontal half-power beam width of the transducer at 4 kHz is 101 degrees with a directivity index of 4 dB. This transmitter can be used for oceanographic applications such as subbottom profiling as well as long-range underwater communication.