Differential expression of flavonoid 3'-hydroxylase during fruit development establishes the different B-ring hydroxylation patterns of flavonoids in Fragaria × ananassa and Fragaria vesca.
Flavonoid 3'-hydroxylase (F3'H) was studied for the first time in different Fragaria species. The cDNA clones isolated from unripe and ripe fruits of Fragaria x ananassa (garden strawberry) and Fragaria vesca (wild strawberry) showed high similarity (99% at the amino acid level) to the publically available F. vesca genome sequence and no significant differences could be identified between species and developmental stages of the fruits. In contrast, the genomic F3'H clones showed differences in the non-coding regions and 5'-flanking elements. The recombinant F3'Hs were functionally active and showed high specificity for naringenin, dihydrokaempferol, and kaempferol, whereas apigenin was only a minor substrate. During fruit development, a clear difference in the F3'H expression was observed between F. × ananassa and F. vesca. While a drastic decline of F3'H expression occurred during fruit ripening in F. × ananassa, F3'H in F. vesca was highly expressed in all stages. This was reflected by the anthocyanin composition, which showed a prevalence of pelargonidin in ripe fruits of F. × ananassa, whereas F. vesca had a high content of cyanidin. Screening of 17 berry species for their anthocyanin and flavonol composition showed that the prevalence of monohydroxylated anthocyanins makes garden strawberry unique among all other fruit species indicating that selection of bright red color during strawberry breeding, which consumers typically associate with freshness and ripeness, has selected phenotypes with a special biochemical background.


Differential penetration of DSIP peptides into rat brain.
Delta sleep-inducing peptide (DSIP) or five closely related peptides were injected peripherally and the levels of DSIP-like immunoreactivity measured in the brains of 100 g and 500 g rats decapitated a minute later after washout with 0.9% NaCl. Higher concentrations of immunoreactive material were found in brain tissue with injection of des Trp1-DSIP, D-Ala4-DSIP, and, after correction for cross-reactivity, with D-Ala4-DSIP-NH2 than with the other peptides. Both peptides substituted in the fourth position with D-alanine resulted in higher concentrations in 500 g rats than in 100 g rats, indicating a possible effect of age. The 5-10 fold greater brain to blood ratios of radioactivity after 125I-N-Tyr-DSIP than after 125-I radioiodinated serum albumin (RISA) provided additional evidence against non-specific leakage or contamination of the brains with substantial amounts of residual blood. The results suggest that penetration of the blood-brain barrier (BBB) by small amounts of peptides, at least DSIP peptides, may be somewhat selective.


Risk Factors for Voriconazole-Associated Hepatotoxicity in Patients in the Intensive Care Unit.
To determine the incidence of hepatotoxicity in critically ill patients who were treated with voriconazole and to identify potential risk factors for voriconazole-associated hepatotoxicity in these patients. Single-center prospective observational study. Intensive care unit (ICU) in a university-affiliated hospital in Xi'an, China. Sixty-three adults, admitted to the ICU between January 2010 and July 2015, who had an ICU length of stay longer than 3 days, had received voriconazole treatment for at least 3 days, and had at least one trough voriconazole plasma concentration (VPC) measurement. All patients received CYP2C19 genotyping and were evaluated for the development of hepatotoxicity by assessing liver function tests performed before, during, and after voriconazole therapy. Hepatotoxicity was classified according to the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) grade scores and was defined as a CTCAE grade score that had increased by at least 2 grade scores over the baseline score. Hepatotoxicity occurred in 12 (19%) of the 63 patients. Characteristics of the patients who developed hepatotoxicity were compared with those of the patients who did not develop hepatotoxicity by univariate and multivariate Cox regression analyses. In the univariate analysis, Acute Physiology and Chronic Health Evaluation II score, invasive fungal infection classification, CYP2C19 genotype, and trough VPC were identified as the variables, and they were subsequently combined in the multivariate regression analysis. Multivariate Cox regression analysis revealed that hepatotoxicity was independently associated with trough VPC (hazard ratio 1.76, p<0.001). The relationships between trough VPCs and probability of hepatotoxicity were explored by using logistic regression analysis, and a target VPC upper limit of 4 mg/L was identified. The Kaplan-Meier method for the cumulative incidence of hepatotoxicity showed a significant difference between patients with trough VPCs of 4 mg/L or higher and those with VPCs lower than 4 mg/L (p<0.001). Trough VPC was an independent risk factor associated with a greater risk of developing hepatotoxicity in critically ill patients, with a potentially toxic target trough concentration threshold of 4 mg/L identified for this complex population.


Coupled cytoplasmic transcription-and-translation--a method of choice for heterologous gene expression in Xenopus oocytes.
We demonstrate here that the intracellular environment of Xenopus oocytes is quite compatible with the requirements of T7 RNA polymerase (T7 RNAP)-mediated transcription. This reaction runs robustly in the oocyte cytoplasm for many hours. The coinjection of a T7 promoter-driven luciferase-encoding plasmid DNA and purified T7 RNAP into oocytes results in the prolonged production of luciferase protein. Thus, the efficient coupling of T7 RNAP-mediated transcription with the intrinsic oocyte translation machinery occurs in the oocyte cytoplasm. The coupled protein synthesis generates high expression yield, displays little variation in the expression level between individual oocytes, requires very limited amounts of DNA template and T7 RNAP, and does not affect the oocyte viability and functional status. Our detailed, quantitative comparison of the existing expression methods in Xenopus oocytes highlights the advantages of the technique based on the cytoplasmic coinjection of T7 RNAP and T7 promoter-driven plasmid DNA and demonstrates that it is greatly superior to the alternative methods of heterologous gene expression.


Combined intracranial Acriflavine, temozolomide and radiation extends survival in a rat glioma model.
Glioblastomas have been historically difficult to treat with poor long-term survival. With novel strategies focused on targeting hypoxia-inducible factor (HIF) regulatory pathways, recent evidence has shown that Acriflavine (ACF) can effectively target glioma invasiveness and recurrence. However, local delivery of ACF and its combinatory effects with Temozolomide (TMZ) and radiation therapy (XRT) have not yet been optimized. In this study we test a novel polymeric matrix that can gradually release ACF at the tumor bed site in combination with systemic TMZ and XRT. In vitro cytotoxicity assays of ACF in combination with TMZ and XRT were performed on rodent and human cell lines with CCK-8 and flow cytometry. In vitro drug release was measured and intracranial safety was assessed in tumor-free animals. Finally, efficacy was assessed in an intracranial gliosarcoma model and combination therapy with TMZ and XRT evaluated. Combination therapy of ACF, TMZ, and XRT was able to reduce cell viability and induce apoptosis in glioma cells. In vitro and in vivo release of ACF was measured in benchtop and animal models. Efficacy was established in an in vivo gliosarcoma model in which intracranial ACF (p < 0.01) significantly improved median survival and the combination therapy of ACF, TMZ and XRT (p < 0.01) significantly improved median survival and led to long-term survival (LTS). We provide evidence that ACF, combined with TMZ and XRT, led to LTS in an intracranial model of rat gliosarcoma. These findings, in combination with the use of a novel polymeric matrix that allows more gradual drug delivery, constitute a first step in the translation of this novel strategy to human use.


Expressional Localization and Functionally Identifying an RNA Editing Enzyme BmADARa of the Silkworm Bombyx mori.
The most common type of RNA editing in metazoans is the deamination of adenosine into inosine (A-to-I) catalyzed by the adenosine deaminase acting on the RNA (ADAR) family of proteins. The deletion or dysfunction of ADAR enzymes in higher eukaryotes can affect the efficiency of substrate editing and cause neurological disorders. However, the information concerning A-to-I RNA editing and ADAR members in the silkworm, Bombyx mori (BmADAR), is limited. In this study, a first molecular comprehensive cloning and sequence analysis of BmADAR transcripts was presented. A complete open reading frame (ORF) (BmADARa) was obtained using RT-PCR and RACE and its expression pattern, subcellular localization and A-to-I RNA-editing function on the silkworm synaptotagmin I (BmSyt I) were investigated. Subcellular localization analysis observed that BmADARa was mainly localized in the nucleus. To further study the A-to-I RNA-editing function of BmADARa, BmSyt I-pIZ-EGFP was constructed and co-transfected with BmADARa-pIZ-EGFP into BmN cells. The result demonstrates that BmADARa can functionally edit the specific site of BmSyt I. Taken together, this study not only provides insight into the function of the first ADAR enzyme in B. mori, but also lays foundations for further exploration of the functional domain of BmADARa and its editing substrates and target sites.


[Parent artery occlusion for the treatment of vertebral artery aneurysm].
To describe the endovascular treatment of posterior aneurysm with balloon occlusion of ipsilateral vertebral artery. 10 patients aging from 12 to 52 years old with mean age 41 were performed occlusion of unilateral vertebral artery after 30 minutes test occlusion. The whole procedure was under systemic anticoagulation. The Willi's Circle was evaluated by MRA before treatment. No immediate complication occurred after the treatment. 10 patients were followed up for 3 months to 5 years. 8 of the 10 cases accepted MR or DSA examination. Focal infarction was found in one patient. The aneurysm was filling in contralateral vertebral artery angiography in one patient, but there was no symptom. Vertebral artery occlusion is an effective treatment to some complicated posterior circulation aneurysms, which is difficult to be clipped and coiled.


The reduced extracellular calcium requirement for proliferation by neoplastic hepatocytes.
Cells from neonatal rat livers were unable to maintain DNA-synthetic activity in calcium-deficient medium, but neoplastic hepatocytes from Morris hepatomas 5123 tc and 7795 synthesized DNA and proliferated indefinitely in this calcium-deficient medium. The calcium content of fresh hepatoma tissue from which these cultures were derived was as much as 10 times greater than that of normal liver; but this difference could not account for the insensitivity of neoplastic cells to extracellular calcium because it disappeared during subsequent cultivation in vitro.


Library collection of the psychosocial publications in consultation-liaison psychiatry.
Psychiatric oncology is a relatively new area, and few comprehensive reviews of related subject matter are available. Thus, the psychiatrists at North Carolina Memorial Hospital who provide liaison to oncology saw the need for a collection of pertinent psychiatric oncology materials that could be studied by faculty, residents, and staff. Such materials would be valuable for patient consultation, lecture preparation, and research background. Their bibliography and library system are discussed here, with a description of the methods of its compilation. The unique features are referenced materials are filed in a centrally available location (departmental library), so that readers have immediate access to them, and bibliographic entries are stored in a small computer in the library, and can be retrieved by topic, title, author. The file currently holds over 340 articles, and is continuously updated. It has been extensively utilized by professionals in this tertiary care teaching hospital as well as in programs across the state.


Assessment of fine needle aspiration cytology samples for molecular genetic analysis in neuroblastoma.
To assess the utility of fine needle aspiration cytology (FNAC) samples for molecular genetic analysis of neuroblastoma. The case files from the pediatric solid tumor clinic were reviewed to identify 20 neuroblastoma patients whose pre-treatment FNAC slides were preserved in the cytology laboratory. The FNAC slides were destained, air dried and hybridisation with fluorescence in situ hybridisation (FISH) probes was performed as per protocol. All slides were screened and analyzed carefully under the fluorescent microscope. Over four-fold increase of the N-myc signal numbers was defined as N-myc amplification. Focal occurrence of cells (at least 50) showing N-myc amplification surrounded by non-amplified tumor cells was defined as focal N-myc amplification. Presence of three or more signals for the long arm of chromosome 17 was defined as 17q gain. FISH analysis gave informative results for all the FNAC smears in our study. FISH analysis of FNAC smears showed N-myc amplification in 5 (25 %) out of 20 patients and 15 (75 %) showed normal N-myc copy number. Three out of these five patients had homogenous amplification and two patients had focal N-myc amplification, indicating tumor heterogeneity. On investigation of chromosome 17q status, 5 (25 %) out of 20 patients demonstrated gain of 17q and 15 (75 %) patients showed normal 17q status. Four out of the five patients with 17q gain also showed N-myc amplification. The current study indicates that FNAC is a rapid and atraumatic diagnostic method for neuroblastoma which provides sufficient material for molecular genetic analyses by means of FISH.


Financial Burden of Hepatocellular Carcinoma Screening in Patients with Cirrhosis.
The overall value of hepatocellular carcinoma screening is defined by the balance of benefits and harms. Studies have only reported physical harms with none describing financial harms. We conducted a multicenter pragmatic randomized clinical trial of hepatocellular carcinoma screening outreach among 2872 patients with cirrhosis from March 2018 to April 2021. Patients with positive or indeterminate results and matched patients with negative results completed surveys at baseline and at follow-up measuring financial harms via Cancer Self-Administered Questionnaire and financial burden via Comprehensive Score for Financial Toxicity Functional Assessment of Chronic Illness Therapy. Univariable and multivariable longitudinal regression analyses were performed to compare changes in financial harms across groups: true positive, true negative, false positive, and indeterminate. Semistructured interviews were conducted in a subset of patients, sampled by center and test result. Of 311 patients who completed at least 1 follow-up survey (75% response rate), 37 had true positive, 133 true negative, 64 false positive, and 77 indeterminate results. Financial harms increased in true positive and false positive patients with no significant changes noted among those with true negative or indeterminate results. At follow-up, 21.8% of patients reported moderate-severe financial burden, which was not significantly associated with test results. Semistructured interviews revealed variation in the frequency and severity of financial harms based on test results, with increased harm in those with false positive results. Financial harms of hepatocellular carcinoma screening vary by test result and can pose a barrier that must be considered when determining the optimal screening program.


Ventricular tachycardia as a consequence of triggered activity.
One of the less frequent underlying mechanisms of ventricular tachycardia (VT) is triggered activity. Triggered activity refers to an extrasystole due to a premature depolarization that occurs when the amplitude of an early or delayed afterdepolarization brings the cardiac membrane to its threshold potential. Hydrochlorothiazide and hydroxyzine can prolong repolarization and QT interval and are associated with early afterdepolarizations. Cyclic AMP-mediated, delayed afterdepolarizations can occur as a result of catecholaminergic surge. Delayed afterdepolarization is classically associated with outflow tract (OT) tachycardia, a type of VT that is uniquely defined by its termination with adenosine. We present a case of triggered OT tachycardia for which intravenous amiodarone through its antiadrenergic effect may have been effective. Infusions of magnesium and a cardioselective, β-receptor antagonist that does not prolong repolarization may have been more appropriate given the concurrent, acquired prolonged QT syndrome. After initial stabilization, considering the underlying VT mechanism may prompt the clinician to select the most appropriate, further treatment.


Regulation of stimulus-induced calcium transport pathways in human T (Jurkat) lymphoblasts.
In human T (Jurkat) lymphoblasts we have studied the calcium signals induced by monoclonal antibodies reacting with the T-cell antigen receptor complex (TCR and CD3). Jurkat cells were preloaded with the fluorescent calcium indicator Indo-1 and the stimulus-induced rise in cytoplasmic free calcium concn was followed in the absence or in the presence of external calcium. The technique allowed the separate investigation of the intracellular calcium release and the external calcium influx processes. The changes in the membrane potential of Jurkat cells were followed simultaneously by using fluorescent indicators. We found that the activation of protein kinase C by phorbol ester (PMA) or by the permeable diacyl glycerol, DiC8, rapidly eliminated the calcium signal, independently of the presence or absence of external calcium, while these treatments did not appreciably change the membrane potential. In contrast, cell membrane depolarization achieved by various treatments selectively blocked the stimulus-induced calcium influx, while did not affect stimulus-induced calcium release from internal stores. The magnitude of the stimulus-induced calcium influx was found to be largely independent of the external calcium concns between about 2-2500 microM. It is demonstrated that the inhibitory effect of membrane depolarization on calcium influx is not simply due to the reduction of the inward calcium gradient under these conditions. These observations indicate a significant down-regulation of the stimulus-induced calcium signal by protein kinase C activation and a selective inhibition of the receptor-operated calcium channels by membrane depolarization.


A picture is worth a thousand words: linking fibromyalgia pain widespreadness from digital pain drawings with pain catastrophizing and brain cross-network connectivity.
Pain catastrophizing is prominent in chronic pain conditions such as fibromyalgia and has been proposed to contribute to the development of pain widespreadness. However, the brain mechanisms responsible for this association are unknown. We hypothesized that increased resting salience network (SLN) connectivity to nodes of the default mode network (DMN), representing previously reported pain-linked cross-network enmeshment, would be associated with increased pain catastrophizing and widespreadness across body sites. We applied functional magnetic resonance imaging (fMRI) and digital pain drawings (free-hand drawing over a body outline, analyzed using conventional software for multivoxel fMRI analysis) to investigate precisely quantified measures of pain widespreadness and the associations between pain catastrophizing (Pain Catastrophizing Scale), resting brain network connectivity (Dual-regression Independent Component Analysis, 6-minute multiband accelerated fMRI), and pain widespreadness in fibromyalgia patients (N = 79). Fibromyalgia patients reported pain in multiple body areas (most frequently the spinal region, from the lower back to the neck), with moderately high pain widespreadness (mean ± SD: 26.1 ± 24.1% of total body area), and high pain catastrophizing scale scores (27.0 ± 21.9, scale range: 0-52), which were positively correlated (r = 0.26, P = 0.02). A whole-brain regression analysis focused on SLN connectivity indicated that pain widespreadness was also positively associated with SLN connectivity to the posterior cingulate cortex, a key node of the DMN. Moreover, we found that SLN-posterior cingulate cortex connectivity statistically mediated the association between pain catastrophizing and pain widespreadness (P = 0.01). In conclusion, we identified a putative brain mechanism underpinning the association between greater pain catastrophizing and a larger spatial extent of body pain in fibromyalgia, implicating a role for brain SLN-DMN cross-network enmeshment in mediating this association.


Bioaccessibilities of metal(loid)s and organic contaminants in particulates measured in simulated human lung fluids: A critical review.
Particle-bound pollutants can pose a health risk to humans. Inhalation exposure evaluated by total contaminant concentrations significantly overestimates the potential risk. To assess the risk more accurately, bioavailability, which is the fraction that enters into the systemic circulation, should be considered. Researchers have replaced bioavailability by bioaccessibility due to the rapid and cost-efficient measurement for the latter, especially for assessment by oral ingestion. However, contaminants in particulates have different behavior when inhaled than when orally ingested. Some of the contaminants are exhaled along with exhalation, and others are deposited in the lung with the particulates. In addition, a fraction of the contaminants is released into the lung fluid and absorbed by the lung, and another fraction enters systemic circulation under the action of cell phagocytosis on particulates. Even if the release fraction, i.e., release bioaccessibility, is considered, the measurement faces many challenges. The present study highlights the factors influencing release bioaccessibility and the incorporation of inhalation bioaccessibility into the risk assessment of inhaled contaminants. Currently, there are three types of extraction techniques for simulated human lung fluids, including simple chemical solutions, sequential extraction techniques, and physiologically based techniques. The last technique generally uses three kinds of solution: Gamble's solution, Hatch's solution, and artificial lysosomal fluid, which are the most widely used physiologically based simulated human lung fluids. External factors such as simulated lung fluid composition, pH, extraction time, and sorption sinks can affect release bioaccessibility, whereas particle size and contaminant properties are important internal factors. Overall, release bioaccessibility is less used than bioaccessibility considering the deposition fraction when assessing the risk of contaminants in inhaled particulates. The release bioaccessibility measurement poses two main challenges: developing a unified, accurate, stable, simple, and systematic biologically based method, and validating the method through in-vivo assays.


How do in-vitro release profiles of nanosuspensions from Alzet® pumps correspond to the in-vivo situation? A case study on radiolabeled fenofibrate.
In research and development sufficiently high and constant plasma levels of drug candidates are often requested, but simple solutions of hydrophobic drugs delivered from the commonly used micro-osmotic pumps cannot meet these demands. Nanosuspensions released from implanted osmotic devices can be a strategy to overcome this challenge but little is known about their pharmacokinetic behavior after subcutaneous application. In the current study, four different nanosuspension formulations containing iodinated fenofibrate were prepared, physicochemically characterized and investigated concerning their in-vitro release kinetics from osmotic pumps. One nanosuspension of lower viscosity exhibited thereby an unexpectedly first order release kinetics, whereas the higher viscous counterpart was released in the expected zero-order manner. To assess the relation of the in-vitro release kinetics to the in-vivo fate of nanosuspensions, various [(131)I] iodinated fenofibrate formulations were subcutaneously applied to mice. The biodistribution was followed by means of γ-scintigraphy and γ-scintillation. Two different nanosuspensions released from osmotic pumps were compared to bolus injections of a nanosuspension and an organic drug solution. The distribution and elimination of the bolus injected drug solution were almost completed within 48h. In contrast, a long lasting (>1week) depot at the injection site was formed by the bolus injected nanosuspension. Ex vivo examination of the organs showed a sustained, but exponential decrease of the radiolabel concentration. More constant drug levels in the organs were achieved within the nanosuspensions released from osmotic pumps. The organ levels of [(131)I] labeled fenofibrate were found to be more constant in case of the pump with the higher viscous nanosuspension in contrast to the lower viscous counterpart. However, the very different release profiles of the lower and higher viscous nanosuspension observed in-vitro were not observed in-vivo, as both pumps showed zero order release. In conclusion, nanosuspensions of poorly soluble compounds released from subcutaneously implanted osmotic pumps can be a suitable approach in pharmacokinetic studies. Although the in-vivo release of nanosuspensions differed in the expected release profile from the in-vitro test results, these in-vitro release tests present a valuable tool for the pre-selection of suitable nanosuspension candidates.


Modulation of amyloid-beta aggregation and toxicity by inosose stereoisomers.
Amyloid-beta (Abeta) aggregation and amyloid formation are key pathological features of Alzheimer's disease, and are considered to be two of the major contributing factors to neurodegeneration and dementia. Identification of small molecule inhibitors that are orally available, have low toxicity and high central nervous system bioavailability is one approach to the potential development of a disease-modifying treatment for Alzheimer's disease. We have previously identified inositol stereoisomers as exhibiting stereospecific inhibition of Abeta aggregation and toxicity in vitro and in vivo. We report here the effects of inosose versus inositol stereoisomers on Abeta fibrillogenesis as determined using CD and fluorescence spectroscopy and negative-stain electron microscopy. The inososes differ from inositols by the oxidation of one of the hydroxyl groups to a ketone. These molecules help in the further elucidation of the structure-activity relationships of inositol-Abeta interactions and identify both allo-inositol and epi-2-inosose as in vitro inhibitors of Abeta aggregation.


[Giardia duodenalis prevalence and associated risk factors in preschool and school-age children of rural Colombia].
Giardia doudenalis infection remains an important public health problem worldwide, as well as in Colombia. This infection is associated with poverty and lack of public services. The study was designed to describe prevalence trends of Giardia duodenalis infections in preschool and school-age children from the village of La Virgen, Quipile, Cundinamarca, and its possible association with socioeconomic variables. Three cross-sectional studies were undertaken in 1995, 2001 and 2005. In each, a non-random sample was selected among children aged <1 to 15 years. Fecal samples were collected for direct examination in order to determine G. duodenalis infection and other intestinal parasites. During the 2005 survey information was collected concerning the sociodemographic and economic conditions in each family group from which a child was sampled. The prevalence of G. duodenalis infection did not change significantly during the study period-13.6% in 1995, 12.8% in 2001 and 15.2% in 2005. These results suggested that belonging to the subsidized health care regime was associated with G. duodenalis infection (prevalence rate [PR]: 4.47; 95% confidence interval [95% CI] 0.6-32.0), not having water supply (PR 2.6; 95% CI 1.1-6.0) and involvement in agricultural activities (PR 2.5; 95% CI 1.0-6.4). Giardiasis is a continuing public health problem at La Virgen. As long as the environmental and socioeconomic factors associated with infection persist, this intestinal protozoan infection will not be controlled.


The management of severe preeclampsia with intravenous magnesium sulphate, hydralazine and central venous catheterization.
A prospective study of patients with severe preeclampsia managed by magnesium sulphate infusion for eclampsia prophylaxis, hydralazine infusion for vasodilatation and central venous catheterization for fluid monitoring is reported. In this preliminary report of 46 patients, there was no episode of eclampsia following treatment, no maternal death, and hypertension was well controlled in all patients. Of the 51 babies born, there were 7 stillbirths, and 4 neonatal deaths; 6 of the 7 stillbirths were less than 30 weeks' gestation, and 23 of the 46 patients were delivered by Caesarean section.


Immunochemical analysis of extracellular matrix during embryonic lens development of the Cat Fraser mouse.
We have characterized the extracellular matrix present during early mouse-lens morphogenesis in Swiss and Cat Fraser mutant mice which produces a thicker capsule. In the two mouse strains, laminin was first detected when the optic vesicle and the head ectoderm are closely associated. At day 10, staining for laminin and fibronectin is especially concentrated at the border of the lens pit. At this stage, type IV collagen and proteoheparan sulphate have a similar distribution to laminin and fibronectin. In the two mouse strains, no major differences were observed in the intensity and the distribution of fluorescent basement-membrane components. This suggests that the overall increase in capsule thickness of the Cat Fraser mutant is more related to an increased cellular synthesis of capsule than to an abnormal distribution of one or more basement-membrane macromolecules.


Epigenetic regulation of thyroid hormone-induced adult intestinal stem cell development during anuran metamorphosis.
Epigenetic modifications of histones are emerging as key factors in gene regulation by diverse transcription factors. Their roles during vertebrate development and pathogenesis are less clear. The causative effect of thyroid hormone (T3) on amphibian metamorphosis and the ability to manipulate this process for molecular and genetic studies have led to the demonstration that T3 receptor (TR) is necessary and sufficient for Xenopus metamorphosis, a process that resembles the postembryonic development (around birth) in mammals. Importantly, analyses during metamorphosis have provided some of the first in vivo evidence for the involvement of histone modifications in gene regulation by TR during vertebrate development. Furthermore, expression and functional studies suggest that various histone modifying epigenetic enzymes likely participate in multiple steps during the formation of adult intestinal stem cells during metamorphosis. The similarity between intestinal remodeling and the maturation of the mammalian intestine around birth when T3 levels are high suggests conserved roles for the epigenetic enzymes in mammalian adult intestinal stem cell development and/or proliferation.


Late development and fertility of adult worms derived from gamma-irradiated first-stage larvae of Angiostrongylus cantonensis.
Effects of gamma ray irradiation on the first-stage larvae of Angiostrongylus cantonensis were studied. Compared to the non-treated controls, infection of rats with third-stage larvae which developed from irradiated first-stage larvae resulted in reduced recovery rates of adults. There was also a change in the male-female adult worm ratio and a reduction in larval output per female in relation to increasing irradiation dosage. Morphological changes in the adults were noted.


Hemispheric asymmetry in reaction time to color stimuli.
The simple detection of colored stimuli in the right, center, and left visual fields was examined. 10 male subjects were used in a reaction time paradigm with color (red, green, or blue), field, and hand as independent variables. A significantly faster RT to stimuli presented in the left visual field was observed, and further the left visual field-left hand combination was the fastest of all the combinations of visual-field x hand. A significant interaction of color x field suggested that red may be processed on a higher order level even in a simple detection task.


A vancomycin-dependent VanA-type Enterococcus faecalis strain isolated in Japan from chicken imported from China.
The characterization of KC122.1, which is a vancomycin-dependent VRE (Vancomycin-resistant enterococci) (Enterococcus faecalis) and the first case in Japan of a VRE isolate obtained from chicken meat imported from China. PCR amplification of vanA, vanS and ddl gene and direct sequencing of the PCR products were performed. KC122.1 was a VanA-type VRE showing high-level vancomycin resistance and low-level teicoplanin resistance, and its vanS gene had three point mutations. The ddl gene of KC122.1 was sequenced and two changes were found at the ninth codon (GCC-GAC) and the stop codon (TAA-CAA). The latter change was also found in the laboratory strain E. faecalis FA2-2. Three point mutations in vanS resulted in high-level vancomycin resistance and low-level teicoplanin resistance. The change at the ninth codon resulted in the inactivation of the ddl gene and vancomycin-dependent growth. An eight amino acid extension at the C-terminal did not impair the function of the D-Ala : D-Ala ligase. This is the first example of the isolation of VRE from chicken meat imported from China and the first vancomycin-dependent VRE from a nonhuman source.


An Alternative Method for Midvault Restoration: "T-Splay Graft".
The internal nasal valve is one of the highly important anatomical points in rhinoplasty procedures. As a key anatomical area, the anatomical and functional integrity of this region must be preserved or reconstructed during rhinoplasty procedures. Several techniques have been defined in the literature for midvault restoration, among which, the most common are spreader grafts and flaps. Both techniques achieve a natural and harmonic contour but may fail to provide the splay effect to the upper lateral cartilages in some cases. A new technique known as the "T-splay graft", to be used as an alternative approach in cases where there is a risk of midvault collapse, and in which the splay effect in the upper lateral cartilage is preferred to be augmented, is explained in the present study. In the present study, a septal tunnel is opened of sufficient size for the cartilage graft planned for placement, at the planned level of the septum, to expand the midvault region and create a splay effect. A bilateral pocket is created in the inferior mucosa of the upper lateral cartilages. The harvested cartilage graft is passed through the septum in the horizontal plane. The cartilage graft is inserted into the bilateral pockets so as way to create convexity to the dorsum. As a result of the applied method, it was established that the internal nasal valve angle could be expanded to the desired extent and the dorsal aesthetic lines could be achieved in a normal anatomical structure. This technique enables the lateral direction of the tension between the graft and the upper lateral cartilages, and the creation of a splay effect. As such, we believe that this method is a highly effective alternative for use in appropriate cases. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Transoral robotic surgery vs. radiotherapy for management of oropharyngeal squamous cell carcinoma - A systematic review of the literature.
Intensity-modulated radiation therapy (IMRT) with or without concurrent chemotherapy is widely utilized for the treatment of oropharyngeal squamous cell carcinoma (OPSCC). However, due to significant acute and late toxicities there has been increasing interest in minimally invasive surgical approaches, particularly transoral robotic surgery (TORS) in an attempt to preserve patient quality of life while maintaining oncologic outcomes. The aim of this study was to review the current literature in order to compare primary IMRT versus TORS in the management of OPSCC. A MEDLINE search was conducted to identify studies reporting on the outcomes of TORS or IMRT in the treatment of OPSCC. Reference lists were also reviewed for relevant articles. Oncologic, functional, and quality of life data is summarized and discussed. One hundred-ninety papers were identified through the MEDLINE search. An additional 52 papers were retrieved by hand searching the reference lists. Ultimately, 44 papers were identified that discussed outcomes after IMRT or TORS for OPSCC. No outcomes from randomized trials were identified. No randomized trials comparing TORS versus IMRT to each other were identified. Uncontrolled reports from the current literature suggest comparable oncologic outcomes with TORS compared to IMRT and functional outcomes may be superior. However, the median follow-up was relatively short and the TORS studies included patients with earlier stage OPSCC on average compared to IMRT studies. Prospective, randomized controlled trials and direct, well-matched comparisons are needed to further elucidate the role for TORS in the treatment of oropharyngeal squamous cell carcinoma.


Investigations on gastroprotective effect of citalopram, an antidepressant drug against stress and pyloric ligation induced ulcers.
The present study investigates the gastroprotective effect of citalopram, an antidepressant drug. Gastroprotective activity of citalopram (5, 10 and 20 mg/kg, bid, po) was evaluated both by single and 14 days repeated pretreatment in the cold restraint stress (CRS) model and 14 days repeated pretreatment in pyloric ligation (PL) model. In addition to ulcer scoring and its histological assessment, levels of corticosterone, hexosamine, nitrite, PGE(2), lipid peroxide and microvascular permeability were also estimated. Mechanism underlying gastroprotective activity was further explored by investigating the involvement of nitric oxide (NO), sulfhydryl (SH) compounds, ATP-sensitive K(+) channels (K(ATP) channels) and prostaglandins (PGs). Results show that against CRS model, repeated pretreatment with citalopram exhibit a significant gastroprotective effect while single pretreatment was ineffective. In CRS model, citalopram repeated pretreatment, in contrast to its single pretreatment, attenuates the corticosterone level and also mitigates the stress-induced increase in nitrite level, lipid peroxidation and microvascular permeability. Additionally, the repeated pretreatment increases the hexosamine and PGE(2) level in CRS model. This gastroprotective effect of citalopram was found to be decreased with L-NAME, NEM, glibenclamide and indomethacin pretreatment. Thus, gastroprotective activity of citalopram appears to be mediated by endogenous NO, SH, PGs and K(ATP) channel opening. In contrast to CRS model, repeated pretreatment with citalopram was ineffective in reducing ulcer formation in PL model.


Predictors and moderators of response to internet-delivered Interpersonal Psychotherapy and Cognitive Behavior Therapy for depression.
By identifying which predictors and moderators lead to beneficial outcomes, accurate selection of the best initial treatment will have significant benefits for depressed individuals. An automated, fully self-guided randomized controlled internet-delivered noninferiority trial was conducted comparing two new interventions (Interpersonal Psychotherapy [IPT; n=620] and Cognitive Behavioral Therapy [CBT; n=610]) to an active control intervention (MoodGYM; n=613) over a period of 4 weeks to spontaneous visitors of an internet-delivered therapy website (e-couch). A range of putative predictors and moderators (socio-demographic characteristics [age, gender, marital status, education level], clinical characteristics [depression/anxiety symptoms, disability, quality of life, medication use], skills [mastery and dysfunctional attitudes] and treatment preference) were assessed using internet-delivered self-report measures at baseline and immediately following treatment and at six months follow-up. Analyses were conducted using Mixed Model Repeated Measures (MMRM). Female gender, lower mastery and lower dysfunctional attitudes predicted better outcome at post-test and/or follow-up regardless of intervention. No overall differential effects for condition on depression as a function of outcome were found. However, based on time-specific estimates, a significant interaction effect of age was found. For younger people, internet-delivered IPT may be the preferred treatment choice, whereas older participants derive more benefits from internet-delivered CBT programs. Although the sample of participants was large, power to detect moderator effects was still lacking. Different e-mental health programs may be more beneficial for specific age groups. The findings raise important possibilities for increasing depression treatment effectiveness and improving clinical practice guidelines for depression treatment of different age groups.


NOx absorption and conversion by ionic liquids.
Ionic liquids (ILs) can be used as absorbents and catalysts for NOx absorption and conversion due to their low toxicity, low energy consumption and excellent reusability. The capacity and absorption mechanism of NOx absorption by ILs are presented in this paper. Generally, NOx are physically absorbed by conventional ILs such as imidazolium-based ILs. The absorption capacity is as follows: NO2>NO>N2O, which is in good agreement with the binding energy between NOx and ILs. Furthermore, low temperature, high pressure and large cation volume are favorable for NOx absorption. The strategies of enhancing NOx capacity through functionalized ILs with metal-containing anions (e.g. [FeCl4]2-), amine groups, sulfonate and carboxylate anions are also concluded. Active N or O sites in functionalized ILs can react with the dimer of NO (N2O2), resulting in high capacity. Moreover, introducing electron-withdrawing substituents such as chlorine and bromine into carboxylate or sulfonate anions reduces desorption residue. Besides NOx absorption, ILs with [NO3]- can activate NO and efficiently catalyze its conversion into HNO3 in the presence of O2 and H2O, and have better performance than ILs with [Cl]-, [Ac]- and [CF3SO3]-, which is attributed to the strong oxidization capability of [NO3]-. In addition, low temperature and high O2 content can further improve NO conversion.


An algorithm for the identification of genetically modified animals.
The diffusion of genetically modified (GM) animals has generated a demand for accurate and unique identification to assure compliance with relevant national and international legislation. Individual identification of GM animals is essential to improve safety and traceability, as well as to fulfill the present and future expectations of producers, consumers, and authorities.


Multiple forms of human kidney mutarotase.
4 forms of mutarotase from human kidney were demonstrated by DEAE-cellulose column chromatography. A major form of them was purified to homogeneity.


Prostaglandin A2 and 35S uptake in connective tissue.
Rats deficient in essential fatty acids (EFA) incorporated lesser amounts of radioactive sulfate into lung, kidney, spleen, heart, costal cartilage, long bone and skull bone than did normal control animals. Administration of prostaglandin A2 stimulated 35S uptake by lung, kidney and aorta while 35S levels in costal cartilage, tibial cap and long bone were strikingly reduced. Comments are presented suggesting that this metabolic mechanism may explain, in part, cartilage and bone resorption in areas of inflammation, such as arthritis, both rheumatoid arthritis and osteoarthritis.


Partial MHC Constructs Treat Thromboembolic Ischemic Stroke Characterized by Early Immune Expansion.
Inflammation and thrombosis are tightly linked, with inflammation contributing to thromboembolism and to stroke outcome. Thromboembolism is a frequent cause of ischemic stroke; yet, the most used occlusion mouse models of experimental stroke do not effectively replicate thromboembolism. Our group recently described a novel thromboembolic mouse model of stroke that successfully occludes the middle cerebral artery with high reproducibility. In the current study, we characterize the peripheral and local immune outcomes as well as the ischemic response to immune therapy in a clinically relevant mouse model of thromboembolic stroke. Brain and spleen tissues were harvested 24 h after thromboembolic stroke and cells immunophenotyped by flow cytometry. We observed a significant increase in neutrophils and early activated T cells in the spleen and an increase in neutrophils and activated monocytes/microglia in the ischemic cortex after thromboembolic stroke. Moreover, as was shown previously for transient MCAO models, treatment of thromboembolic stroke with partial MHC constructs significantly reduced ischemic damage indicating an equivalent effect of this immune-based therapy in the thromboembolic model that better mimics the pathophysiology of human stroke.


Nephrinuria and podocytopathies.
The discovery of nephrin in 1998 has launched a new era in glomerular diseases research, emphasizing its crucial role in the structure and function of the glomerular filtration barrier. In the past 20 years, substantial advances have been made in understanding podocyte structure and function as well as the discovery of several podocyte-related proteins including nephrin. The glomerular filtration barrier is comprised of podocytes, the glomerular basement membrane and endothelial cells. Podocytes, with their specialized slit diaphragm, form the essential backbone of the glomerular filtration barrier. Nephrin is a crucial structural and functional feature of the slit diaphragm that prevents plasma protein, blood cell and macromolecule leakage into the urine. Podocyte damage results in nephrin release. Podocytopathies are kidney diseases in which podocyte damage drives proteinuria, i.e., nephrotic syndrome. Many kidney diseases involve podocytopathy including congenital nephrotic syndrome of Finnish type, diffuse mesangial sclerosis, minimal change disease, focal segmental glomerulosclerosis, collapsing glomerulonephropathy, diabetic nephropathy, lupus nephropathy, hypertensive nephropathy and preeclampsia. Recently, urinary nephrin measurement has become important in the early detection of podocytopathies. In this chapter, we elaborate the main structural and functional features of nephrin as a podocyte-specific protein, pathomechanisms of podocytopathies which result in nephrinuria, highlight the most commonly used methods for detecting urinary nephrin and investigate the diagnostic, prognostic and potential therapeutic relevance of urinary nephrin in primary and secondary proteinuric kidney diseases.


Freshwater insects of different feeding guilds ingest microplastics in two Gulf of Guinea tributaries in Nigeria.
Plastic pollution has enormous impacts on freshwater and marine ecosystem health, and it is one of the topmost environmental concerns of the current geological period (i.e. the Anthropocene). Thus, the goal of our study was to provide baseline information and bridge the information gap on the occurrence of microplastics (MPs) in African freshwater systems, using two tributaries of the Gulf of Guinea (Ogun and Osun Rivers) in Nigeria as a case study and three freshwater insect species of different feeding guilds as bioindicators. A total of 29 individuals of the insect species were chemically digested and subsequently analysed for MP presence under a digital microscope and a micro-Fourier-transform infrared (μFTIR) spectroscope. Collector-gatherers (Chironomus sp. and Siphlonurus sp.) recorded the highest MP load per gram wet weight, while the predatory Lestes viridis recorded the lowest. The highest diversity of polymers was recorded in Chironomus sp. of Ogun River, i.e. styrene ethylene butylene styrene, acrylonitrile butadiene styrene (ABS), chlorinated polyethylene, polypropylene (PP), and polyester, while two polymers each were recorded in Siphlonurus sp. (i.e. polyester and ABS) and L. viridis (i.e. polyester and PP) of Osun River. We conclude that collector-gatherers like Chironomus sp. and Siphlonurus sp. could be best employed as MP bioindicators in freshwater systems. However, their suitability as MP bioindicators should be further investigated in different freshwater ecosystems worldwide. Graphical abstract.


Sylvatic dengue virus type 2 activity in humans, Nigeria, 1966.
Using phylogenetic analysis of complete virus genomes from human isolates obtained in Nigeria in 1966, we identified sylvatic dengue virus (DENV) strains from 3 febrile patients. This finding extends current understanding of the role of sylvatic DENV in febrile disease and documents another focus of sylvatic DENV transmission in West Africa.


The construction and assessment of lenses for local treatment of malignant tumours by ultrasound.
A simple method to build ultrasound lenses from araldite has been developed. The focussing properties of these lenses as a function of sonic frequency and lens diameter were studied using a computer model. The optimum frequency and diameter of these lens were found to be around 1 MHz and between 40 and 60 mm respectively, for deep tissue heating. The stability of the power output as well as the temperature distributions produced in vitro and in vivo have also been studied. It appears to be possible to produce larger temperature elevations near the acoustical focus than near the skin surface in vivo. Therefore the lenses seem to be suitable for local cancer treatment.


Gasification of the char derived from distillation of granulated scrap tyres.
This work reports the effect of pressure on the steam/oxygen gasification at 1000°C of the char derived from low temperature-pressure distillation of granulated scrap tyres (GST). The study was based on the analysis of gas production, carbon conversion, cold gas efficiency and the high heating value (HHV) of the product. For comparison, similar analyses were carried out for the gasification of coals with different rank. In spite of the relatively high ash (≈12 wt.%) and sulphur (≈3 wt.%) contents, the char produced in GST distillation can be regarded as a reasonable solid fuel with a calorific value of 34MJkg(-1). The combustion properties of the char (E(A)≈50 kJ mol(-1)), its temperature of self-heating (≈264°C), ignition temperature (≈459°C) and burn-out temperature (≈676°C) were found to be similar to those of a semi-anthracite. It is observed that the yield, H(2) and CO contents and HHV of the syngas produced from char gasification increase with pressure. At 0.1 MPa, 4.6 Nm(3)kg(char)(-1) of syngas was produced, containing 28%v/v of H(2) and CO and with a HHV around 3.7 MJ Nm(-3). At 1.5 MPa, the syngas yield achieved 4.9N m(3)kg(char)(-1) with 30%v/v of H(2)-CO and HHV of 4.1 MJ Nm(-3). Carbon conversion significantly increased from 87% at 0.1 MPa to 98% at 1.5 MPa. It is shown that the char derived from distillation of granulated scrap tyres can be further gasified to render a gas of considerable heating value, especially when gasification proceeds at high pressure.


Lipids and sterols in Musca domestica L. (Diptera, Muscidae): changes after treatment with sucrose and lead.
Total lipid, fatty acid and sterol composition of larvae and adults of Musca domestica was investigated before and after feeding on sucrose syrup or on the same syrup containing 1% lead nitrate. The effects of sucrose and of lead ions were found to be different. In larvae sucrose diet inhibited the fatty acid elongation and stimulated the first stages of their unsaturation. A significant increase of phytosterol concentrations was obtained. These changes increased the cell membrane permeability. The addition of lead caused a decrease of the fatty acid unsaturation, which decreased the cell membrane permeability. In adults the sucrose diet had no effect on the lipid and sterol composition, while the addition of lead decreased the cholesterol concentration. The composition of lipids and sterols also depends on the diet of larvae before pupation. The data obtained suggested that changes in lipid and sterol composition, which control the permeability of the cell membrane, might be an adaptive response of the organism to the changes of the environment.


Venezuelan encephalitis virus: in vivo induction of a chromosomal abnormality in hamster bone marrow cells.
This study reports the induction in vivo of aneuploidy in bone marrow cells of the Syrian hamster after short-term infection with Venezuelan encephalitis virus.


Whole-body MRI in paediatric oncology.
Imaging plays a crucial role in the diagnosis and follow-up of paediatric malignancies. Until recently, computed tomography (CT) has been the imaging technique of choice in children with cancer, but nowadays there is an increasing interest in the use of functional imaging techniques like positron emission tomography and single-photon emission tomography. These later techniques are often combined with CT allowing for simultaneous acquisition of image data on the biological behaviour of tumour, as well as the anatomical localisation and extent of tumour spread. Because of the small but not negligible risk of radiation induced secondary cancers and the significantly improved overall survival rates of children with cancer, there is an increasing interest in the use of alternative imaging techniques that do not use ionising radiation. Magnetic resonance imaging (MRI) is a radiation-free imaging tool that allows for acquiring images with a high spatial resolution and excellent soft tissue contrast throughout the body. Moreover, recent technological advances have resulted in fast diagnostic sequences for whole-body MR imaging (WB-MRI), including functional techniques such as diffusion weighted imaging. In this review, the current status of the technique and major clinical applications of WB-MRI in children with cancer will be discussed.


Trichocladabiflavone A, a chalcone-flavonone type biflavonoid from Selaginella trichoclada Alsto.
A chalcone-flavonone type biflavonoid, trichocladabiflavone A (1), along with eight known biflavonoids (2-9) were isolated from the 70% EtOH extract of Selaginella trichoclada. Their structures were elucidated by extensive spectroscopic analyses. Compound 1 was the first chalcone-flavonone type biflavonoid reported in the genus Selaginella. Moreover, compound 1 exhibited moderate cytotoxicity against DU145, MCF-7 and PC3 human cancer cell lines.


Defective angiogenesis in the inflammatory granulation tissue in histidine decarboxylase-deficient mice but not in mast cell-deficient mice.
We have analyzed the role of histamine in the angiogenesis of the granulation tissue in histidine decarboxylase-deficient (HDC(-/-)) mice, mast cell-deficient mice (WBB6F1-W/W(V)), and their corresponding wild-type mice (HDC(+/+) and WBB6F(1)(+/+)). In HDC(+/+) mice, subcutaneous implantation of a cotton thread in the dorsum induced granulation tissue formation with angiogenesis, while the topical injection of anti-vascular endothelial growth factor (VEGF) IgG strongly suppressed them. In HDC(-/-) mice which showed lower VEGF levels in the granulation tissue, there was notably less angiogenesis and granulation tissue formation than in HDC(+/+) mice. The topical injection of histamine or the H(2) agonist dimaprit rescued the defective angiogenesis and granulation tissue formation in HDC(-/-) mice. There was no significant difference in the granulation tissue formation and angiogenesis between WBB6F1-W/W(V) and WBB6F1(+/+) mice. In addition, macrophages in the granulation tissue were found to express HDC. Our findings indicate that histamine derived from non-mast cells plays a significant role in the angiogenesis of the inflammatory granulation tissue.


Genome-wide identification and expression analysis of the calmodulin-binding transcription activator (CAMTA) gene family in wheat (Triticum aestivum L.).
Plant calmodulin-binding transcription activator (CAMTA) proteins play important roles in hormone signal transduction, developmental regulation, and environmental stress tolerance. However, in wheat, the CAMTA gene family has not been systematically characterized. In this work, 15 wheat CAMTA genes were identified using a genome-wide search method. Their chromosome location, physicochemical properties, subcellular localization, gene structure, protein domain, and promoter cis-elements were systematically analyzed. Phylogenetic analysis classified the TaCAMTA genes into three groups (groups A, B, and C), numbered 7, 6, and 2, respectively. The results showed that most TaCAMTA genes contained stress-related cis-elements. Finally, to obtain tissue-specific and stress-responsive candidates, the expression profiles of the TaCAMTAs in various tissues and under biotic and abiotic stresses were investigated. Tissue-specific expression analysis showed that all of the 15 TaCAMTA genes were expressed in multiple tissues with different expression levels, as well as under abiotic stress, the expressions of each TaCAMTA gene could respond to at least one abiotic stress. It also found that 584 genes in wheat genome were predicted to be potential target genes by CAMTA, demonstrating that CAMTA can be widely involved in plant development and growth, as well as coping with stresses. This work systematically identified the CAMTA gene family in wheat at the whole-genome-wide level, providing important candidates for further functional analysis in developmental regulation and the stress response in wheat.


A retrospective pilot study examining the use of Acthar gel in sarcoidosis patients.
Acthar was reported as effective for the treatment of pulmonary sarcoidosis in the 1950s. Use of drug waned due to cost and toxicity compared to prednisone. Recent interest has reemerged as an alternative to high dose oral glucocorticoids. Chart review was performed on all advanced sarcoidosis patients seen at two centers who received at least one dose of Acthar gel therapy with at least six months of posttreatment follow up. In all cases prior sarcoidosis therapy and indications for use along with clinical outcome were noted. All patients initially received 80 IU intramuscular or subcutaneous administration twice a week. A total of 47 patients were treated with Acthar gel therapy during the study period, and 18 (37%) discontinued drug within six months due to cost (four patients), death (two patients), or drug toxicity (eleven patients), or noncompliance (1 patient). Of the remaining 29 patients, eleven experienced objective improvement in one or more affected organs. All but two patients noted disease improvement or oral glucocorticoid reduction. Twenty-one patients were treated for more than six months (Median 274 days). Nineteen patients were on prednisone at time of starting Acthar gel: seventeen had their prednisone dosage reduced by more than fifty percent and one patient discontinued cyclophosphamide therapy. In this group of advanced sarcoidosis patients, Acthar gel treatment for at least three months was associated with objective improvement in a third of patients. A third of patients were unable to take at least a three months of treatment.


Dissecting RNA folding by nucleotide analog interference mapping (NAIM).
Nucleotide analog interference mapping (NAIM) is a powerful chemogenetic approach that allows RNA structure and function to be characterized at the atomic level. Random modifications of base or backbone moieties are incorporated into the RNA transcript as nucleotide analog phosphorothioates. The resulting RNA pool is then subjected to a stringent selection step, in which the RNA has to accomplish a specific task, for example, folding. RNA functional groups important for this process can be identified by physical isolation of the functional and the nonfunctional RNA molecules and subsequent mapping of the modified nucleotide positions in both RNA populations by iodine cleavage of the susceptible phosphorothioate linkage. This approach has been used to analyze a variety of aspects of RNA biochemistry, including RNA structure, catalysis and ligand interaction. Here, I describe how to set up a NAIM assay for studying RNA folding. This protocol can be readily adapted to study any RNAs and their properties. The time required to complete the experiment is dependent on the length of the RNA and the number of atomic modifications tested. In general, a single NAIM experiment can be completed in 1-2 weeks, but expect a time frame of several weeks to obtain reliable and statistically meaningful results.


The will of Congress? Permissive regulation and the strategic use of labeling for the anti-influenza drug Relenza.
Through an analysis of the FDA's approval of the controversial anti-influenza drug Relenza (zanamivir), we interrogate distinct social scientific theories of pharmaceutical regulation. We investigate why, despite internal negative opinions and an Advisory Committee's non-approval recommendation, the FDA approved Relenza in the late 1990s. Based on a close reading of FDA documents, we show how agency officials guided the manufacturer's analyses and participated in constructing a tenuous argument for approval. We show how regulators may strategically design drug labels that can justify their permissive regulation. We consider the explanatory power of official accounts and alternative, partially overlapping, theories of pharmaceutical regulation in the Relenza case, and develop new insights into the institutional dynamics of regulator-industry relations. We find little or no evidence that the FDA was primarily driven by public health concerns, pressure from disease-based patient activism, or a consumerist and neoliberal regulatory logic, although some of these explanations provided managers with convenient rhetoric to rationalize their actions. Rather, we argue that the Relenza case highlights contradictions between a scientific culture at FDA, conducive to rigorous product evaluations, and the agency's attempts to accommodate higher-level political (i.e. Congress) and industry demands conducive of permissive regulation - consistent with some aspects of reputational and capture theories, as well as with corporate bias theory.


Mutagenesis of a Gly-Ser cleavage site in MUC1 inhibits ectodomain shedding.
MUC1 mucin is a type 1 transmembrane glycoprotein dimer of extracellular and membrane-bound subunits. The two non-covalently associated subunits are produced from a single polypeptide chain by proteolysis at a Gly-Ser peptide bond in the endoplasmic reticulum prior to localization on the cell surface. However, once expressed on the surface, the extracellular subunit is shed from cells in the absence of the membrane-associated subunit. Previous studies implicated a cellular metalloproteinase mediating MUC1 ectodomain shedding, but no reports have delineated the site of metalloproteinase cleavage or directly assessed the role of the Gly-Ser bond in shedding. Therefore, we performed site-directed mutagenesis of the Gly-Ser site and determined the effects on MUC1 proteolysis and shedding. Ser-->Ala substitution blocked MUC1 cleavage and inhibited shedding. Equal amounts of wild type and mutant MUC1 were expressed on the cell surface, indicating that lack of shedding of the mutant molecule was not due to reduced surface localization. We conclude that the Gly-Ser peptide bond is required for MUC1 shedding.


Using Tailored Videos to Teach Inhaler Technique to Children With Asthma: Results From a School Nurse-Led Pilot Study.
Our purpose was to test whether a tailored inhaler technique video intervention: (1) could be feasibly implemented by school nurses and (2) improve the inhaler technique of children with asthma. School nurses recruited a convenience sample of 25 children with asthma (ages 7-17) and assessed their inhaler technique. Children then watched a tailored video that provided: (1) step-by-step feedback on which steps (out of 8) they performed correctly, (2) praise for correctly-performed steps, and (3) statements about why incorrectly-performed steps are important. Nurses reassessed the child's inhaler technique immediately after watching the video and again 1month later. Non-parametric Wilcoxon signed rank tests were calculated to assess whether children's technique significantly improved from baseline to post-video and baseline to 1-month follow-up. A focus group with the school nurses was conducted post-intervention to discuss feasibility issues. Children's inhaler technique improved by 1.2 steps (with spacer; p=0.03) and 2.7 steps (without spacer; p<0.01) from baseline to post-video. These improvements were maintained at 1-month follow-up. School nurses believed the intervention was feasible to implement and met an important educational need. A school nurse-led tailored video intervention is feasible to implement and a promising method for improving children's inhaler technique.


Nonepileptic seizures in individuals attending neurological services in New Zealand.
This study examined the demographic, social, and clinical (neurological and psychiatric) characteristics of people with psychogenic nonepileptic seizures (PNES) presenting to tertiary neurological services at Auckland District Health Board, New Zealand. Electronic notes and video-electroencephalography (video-EEG) data gathered from the telemetry unit based on synchronized acquisition (motor activity and brain electrophysiology) over a five-year period (2011 to 2015 inclusive) were retrospectively examined. Two groups were compared: people with PNES only or people with combined PNES and epileptic seizures (ES) (the group with PNES) and a control group with ES only, matched 1:1 by age and gender. Sixty-six people in the group with PNES were matched with an equivalent number of ES controls. As a cohort, there was high psychiatric and medical comorbidity in both groups, but overall, those with PNES experienced higher rates than their ES counterparts. An older age of onset, female gender, and history of abuse were more frequently seen in those with PNES. Compared with controls, people with PNES more commonly had daily seizures (rather than monthly) but presented less frequently to neurology services. A high proportion of people with PNES experienced historical traumas, ongoing stressors, and disability. Almost half of the people with PNES were on antiepileptic drugs (AEDs) and received limited psychiatric or psychological input. In contrast, people with ES were more likely to be on psychotropic medication. Both PNES and ES are associated with high levels of psychiatric and medical comorbidity. Additionally, PNES were found to be associated with iatrogenic harm and disability. Though people with PNES and ES often have a range of associated needs, there is, however, limited access to appropriate services. The needs of these populations should be better met through enhanced integration across psychiatry, neurology, and multidisciplinary services.


Prospective multicenter evaluation of a new immunoassay and real-time PCR for rapid diagnosis of Clostridium difficile-associated diarrhea in hospitalized patients.
In a prospective multicenter study, 367 fecal samples from 300 patients with diarrhea were tested for Clostridium difficile-associated diarrhea (CDAD) with a new immunochromatography assay for toxins A and B (ICTAB), a real-time PCR on the toxin B gene, and the cell cytotoxicity assay. Twenty-three (6.2%) of the 367 fecal samples were positive by the cell cytotoxicity assay. With the cell cytotoxicity assay as the "gold standard," the sensitivity, specificity, positive predictive value, and negative predictive value for the ICTAB assay and real-time PCR were 91, 97, 70, and 99%, and 87, 96, 57 and 99%, respectively. In conclusion, both the ICTAB and the real-time PCR can be implemented as rapid screening methods for patients suspected of having CDAD.


Commentary on: Saper CB, Loewy AD, Swanson LW, Cowan WM. (1976) Direct hypothalamo-autonomic connections. Brain Research 117:305-312.
The 1970s saw the introduction of new technologies for tracing axons both anterogradely and retrogradely. These methods allowed us to visualize fine, unmyelinated pathways for the first time, such as the hypothalamic pathways that control the autonomic nervous system. As a result, we were able to identify the paraventricular nucleus and lateral hypothalamus as the key sites that provide direct inputs to the autonomic preganglionic neurons in the medulla and spinal cord. These findings revolutionized our understanding of hypothalamic control of the autonomic nervous system.


2-(4-N-maleimidophenyl)-6-methoxybenzofuran: a superior derivatizing agent for fluorimetric determination of aliphatic thiols by high-performance liquid chromatography.
The use of 2-(4-N-maleimidophenyl)-6-methoxybenzofuran as a fluorogenic pre-column derivatizing agent for sensitive high-performance liquid chromatographic (HPLC) determination of aliphatic thiols is described. The maleamic acid, its methyl ester and the N-maleimide derivatives of the common amine precursor were easily synthesized and characterized according to conventional methods. These derivatives lack fluorescent properties in their native form but display strong fluorescence upon reaction with aliphatic thiols. The reaction is rapid and highly selective for thiols over a wide pH range (7.1-8.8). Following derivatization, the thiol adducts were separated on a reversed-phase column (Ultrasphere-ODS) using 0.1% hexanesulfonic acid in 10 mM potassium hydrogen phosphate-acetonitrile (65:35, pH 4.5) and were detected fluorimetrically (excitation at 310 nm; emission at 390 nm). The method is highly sensitive (femtomole range) and is easily applied to the determination of SH-containing drugs and endogenous thiols in biological samples.


Teratologic studies on the Himalayan rabbit: new aspects of thalidomide-induced teratogenesis.
The aim of our study was to determine the period of maximum sensitivity for the induction of characteristic malformations with thalidomide (TH) in Himalayan rabbits. TH was administered orally in different doses (50, 100, 150 and 200 mg/kg) four times at 24-h intervals starting at 192 h of gestation. The malformations affected various organs: renal defects (dysplasia) and limb anomalies (dysmelia)--which had never occurred spontaneously in this strain--appeared as dose-dependent effects of the drug. By administering single doses of TH (200 and 300 mg/kg body wt) between hours 192 and 264 of gestation, we discovered the different periods of maximum sensitivity for induction of renal dysplasia (clearly prior to the 220th h of gestation) and dysmelia (between hours 230 and 240 of gestation). The types of limb malformations that we observed in the rabbit were identical to those produced in man following the intake of TH. Three doses of TH (300 mg/kg each) given between hours 222 and 228 of gestation produced characteristic limb malformations in 9 of 11 litters treated. These results make it possible to conduct in vivo experiments on a readily available laboratory animal with minor drug exposure of the gravid dam and under avoidance of toxic side effects.


Fault diagnosis and prediction of wind turbine gearbox based on a new hybrid model.
Gearbox is an important part of wind turbine. Diagnosing and forecasting gearbox faults of wind turbines can effectively reduce the costs of operation and maintenance and improve the reliability of gearbox operation. Due to high dimensionality and nonlinearity of system parameters, the paper uses the grey relation analysis to select features related to gearbox oil temperature. Features with a relational degree above 0.7 are selected as input data related to oil temperature, including wind speed, ambient temperature, power, and gearbox shaft temperature. Then, a new extreme learning machine with kernel improved by the whale optimization algorithm is established to forecast gearbox oil temperature. Through the residuals between gearbox oil temperature predicted by the proposed model and monitored by the SCADA, whether the gearbox exists faults can be diagnosed. In the case study, the test data was divided into two groups (the test data with and without faults). In the data test without faults, compared with three other models, the proposed model has the smallest false-negative rate (0.211%) and mean absolute percentage error (2.812%). In the data test with faults, the proposed model can diagnose gearbox faults earlier (160 min in advance) than the other three benchmark models. The results show that the proposed hybrid model performs well in the fault diagnosis and prediction of wind turbine gearbox.


A Long Noncoding RNA Regulates Hepatitis C Virus Infection Through Interferon Alpha-Inducible Protein 6.
Long noncoding RNAs (lncRNAs) play a critical role in the regulation of many important cellular processes. However, the mechanisms by which lncRNAs regulate viral infection and host immune responses are not well understood. We sought to explore lncRNA regulation of hepatitis C virus (HCV) infection and interferon response. We performed RNA sequencing (RNAseq) in Huh7.5.1 cells with or without interferon alpha (IFNα) treatment. Clustered regularly interspaced short palindromic repeats/Cas9 guide RNA (gRNA) was used to knock out selected genes. The promoter clones were constructed, and the activity of related interferon-stimulated genes (ISGs) were detected by the secrete-pair dual luminescence assay. We constructed the full-length and four deletion mutants of an interferon-induced lncRNA RP11-288L9.4 (lncRNA-IFI6) based on predicted secondary structure. Selected gene mRNAs and their proteins, together with HCV infection, in Huh7.5.1 cells and primary human hepatocytes (PHHs) were monitored by quantitative real-time PCR (qRT-PCR) and western blot. We obtained 7,901 lncRNAs from RNAseq. A total of 1,062 host-encoded lncRNAs were significantly differentially regulated by IFNα treatment. We found that lncRNA-IFI6 gRNA significantly inhibited HCV infection compared with negative gRNA control. The expression of the antiviral ISG IFI6 was significantly increased following lncRNA-IFI6 gRNA editing compared with negative gRNA control in Japanese fulminant hepatitis 1 (JFH1)-infected Huh7.5.1 cells and PHHs. We observed that lncRNA-IFI6 regulation of HCV was independent of Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling. lncRNA-IFI6 negatively regulated IFI6 promoter function through histone modification. Overexpression of the truncated spatial domain or full-length lncRNA-IFI6 inhibited IFI6 expression and increased HCV replication. Conclusion: A lncRNA, lncRNA-IFI6, regulates antiviral innate immunity in the JFH1 HCV infection model. lncRNA-IFI6 regulates HCV infection independently of the JAK-STAT pathway. lncRNA-IFI6 exerts its regulatory function via promoter activation and histone modification of IFI6 through its spatial domain.


The NIN transcription factor coordinates CEP and CLE signaling peptides that regulate nodulation antagonistically.
Legumes tightly regulate nodule number to balance the cost of supporting symbiotic rhizobia with the benefits of nitrogen fixation. C-terminally Encoded Peptides (CEPs) and CLAVATA3-like (CLE) peptides positively and negatively regulate nodulation, respectively, through independent systemic pathways, but how these regulations are coordinated remains unknown. Here, we show that rhizobia, Nod Factors, and cytokinins induce a symbiosis-specific CEP gene, MtCEP7, which positively regulates rhizobial infection. Via grafting and split root studies, we reveal that MtCEP7 increases nodule number systemically through the MtCRA2 receptor. MtCEP7 and MtCLE13 expression in rhizobia-inoculated roots rely on the MtCRE1 cytokinin receptor and on the MtNIN transcription factor. MtNIN binds and transactivates MtCEP7 and MtCLE13, and a NIN Binding Site (NBS) identified within the proximal MtCEP7 promoter is required for its symbiotic activation. Overall, these results demonstrate that a cytokinin-MtCRE1-MtNIN regulatory module coordinates the expression of two antagonistic, symbiosis-related, peptide hormones from different families to fine-tune nodule number.


The experience of patients with cancer who develop venous thromboembolism: an exploratory study.
To better understand the experience of venous thromboembolism (VTE) from the points of view of patients with cancer during various stages of the cancer experience. Qualitative, descriptive. Various inpatient and outpatient units of a large urban university-affiliated hospital in Montreal, Quebec, Canada. Purposive sample of 10 participants who were anticipating, had recently undergone, or were currently undergoing cancer treatment and who had received a VTE diagnosis within the past year. Semistructured interviews were transcribed verbatim. Thematic analysis of data revealed themes contributing to understanding the lived experience of VTE during cancer care. The experience of patients with cancer who develop VTE. Patients' initial reaction to VTE included VTE as a life-threat, past experience with VTE, and VTE as the "cherry on the sundae" in light of other cancer-related health issues. Patients' coping with VTE also included three themes: VTE being overshadowed by unresolved cancer-related concerns, VTE as a setback in cancer care, and attitudes about VTE treatment. This study contributes new insight into the experience of patients with cancer who develop VTE. The most salient finding was that patients having no prior VTE knowledge experienced VTE as more challenging. Future studies comparing experiences with VTE across the various stages of cancer care are needed. Study findings suggest that patient education about VTE would be useful for the initial reaction and subsequent coping phases of VTE, thus representing an important target area for nursing intervention.


Apolipoprotein E2, renal failure and lipid abnormalities in non-insulin-dependent diabetes mellitus.
The association of apolipoprotein E (apo E) genetic polymorphism, particularly apo E2, with renal failure (plasma creatinine > or = 1.4 mg/dl, and urinary albumin excretion index > or = 300 mg/g.creatinine and/or persistent proteinuria) was investigated in 57 non-insulin-dependent diabetic (NIDDM) patients. Apo E2 allele frequency was significantly higher in diabetic patients with renal failure (9.6%) than in diabetic patients without renal failure (3.2%) and in the general Japanese population (3.7%). This finding suggests that apo E2 is associated with renal failure in NIDDM. In addition, to elucidate the association of apo E2 with lipid abnormalities, plasma lipid and lipoprotein levels were compared among the apo E2 (E2/2 and E3/2) and E3/3 groups of NIDDM with renal failure (n = 27) and the apo E2 (E3/2) and E3/3 groups of NIDDM with normoalbuminuria (n = 34). In diabetic patients, the apo E2 group with renal failure had significantly higher levels of plasma total cholesterol (T-chol), very-low-density lipoprotein (VLDL)-chol, triglyceride (TG), VLDL-TG and apo E than the apo E3/3 group with renal failure, and had significantly higher levels of plasma T-chol, VLDL-chol, TG and VLDL-TG than the apo E2 and E3/3 groups with normoalbuminuria. Furthermore, the apo E2 group with renal failure had significantly higher ratios of VLDL-(chol/TG) and VLDL-chol/TG (an index of remnants in plasma) than the apo E3/3 group with renal failure and the apo E2 and E3/3 groups with normoalbuminuria. These results suggest that apo E2 leads to the accumulation of TG-rich lipoprotein and remnants in plasma. It is concluded that apo E2 is associated with renal insufficiency in NIDDM and that apo E2 may be a factor that aggravates lipid abnormalities in NIDDM with renal failure.


Utilization of microarrayed compound screening (microARCS) to identify inhibitors of p56lck tyrosine kinase.
Protein tyrosine kinases play critical roles in cell signaling and are considered attractive targets for drug discovery. The authors have applied microARCS (microarrayed compound screening) technology to develop a high-throughput screen for finding inhibitors of the p56lck tyrosine kinase. Initial assay development was performed in a homogeneous time-resolved (LANCE) format in 96-well microplates and then converted into the gel-based microARCS format. The microARCS methodology is a well-less screening format in which 8640 compounds are arrayed on a microplate-sized piece of polystyene and subsequently assayed by placing reagents cast in agarose gels in contact with these compound sheets. A blotting paper soaked with adenosine triphosphate is applied on the gel to initiate the kinase reaction in the gel. Using this screening methodology, 300,000 compounds were screened in less than 40 h. Substantial reagent reduction was achieved by converting this tyrosine kinase assay from a 96-well plate assay to microARCS, resulting in significant cost savings.


Single-stage repair of bladder exstrophy in older children and children with failed previous repair.
Evaluation of cosmetic and functional outcome of single-stage exstrophy-epispadias complex repair in older children and those with previously failed repair. This study comprised 15 children (12 boys and 3 girls) with classic bladder exstrophy and a mean age at repair of 8.6 months (range 2-24 months). Eight children had a previously failed repair. All children underwent complete primary repair using the single-stage Mitchell technique. Half of the boys had complete penile disassembly, while in the others a modified Cantwell-Ransley technique for epispadias repair was used. Anterior iliac osteotomy was performed and hip spica used for immobilization in all children. One child had urethral stricture treated by endoscopic visual urethrotomy. Three children had penopubic fistulae that closed spontaneously. No bladder dehiscence or prolapse was encountered. Vesicoureteral reflux was present in 20 renal units but ureteral reimplantation was not performed. Average bladder capacity after closure was 134 cm(3) (range 110-160 cm(3)) with only two partially continent and six incontinent children. Mean follow-up period is 2 years (range 1-3 years). Single-stage repair was performed in children with previously failed repair and those presenting at an older age with satisfactory results. Acceptable bladder and genital anatomy and function were achieved together with preservation of renal function. The impact of this technique on continence is not encouraging, but needs to be determined in a longer follow-up period.


Autism spectrum disorder and Li-Fraumeni syndrome: purely coincidental or mechanistically associated?
Autism spectrum disorders (ASDs) are neurodevelopmental disorders with impaired social interactions and communication and restrictive, repetitive patterns of behaviors, interests, and activities. A recent epidemiological study suggests that children with ASD might have an increased cancer risk. The 14.5-year-old boy, previously diagnosed with ASD, was referred with persistent bone pain. Diagnostic work-up confirmed diagnosis of acute lymphoblastic leukemia (ALL); cytogenetic analysis revealed low hypodiploid karyotype with a mutation (c.733G>A, p.Gly245Ser, rs28934575) in TP53 in the leukemic blasts. By Sanger sequencing, the presence of this mutation in the germline was subsequently confirmed and, thus, diagnosis of Li-Fraumeni syndrome (LFS) was made. His family history was remarkable with two siblings with intellectual disability and a mother who has died of premenopausal breast cancer. Some of the oncogenes and tumor suppressor genes causing cancer susceptibility syndromes overlap with those involved in autism. This functional overlap between autism and cancer is novel and particularly compelling. The surprising coincidence of LFS and ASD in our patient raises the question whether this is purely coincidental or mechanistically associated.


Comparing non-invasive diagnostic methods for arteriovenous fistula stenosis: a prospective study.
International guidelines recommend screening for arteriovenous fistula (AVF) stenosis using various non-invasive methods. We evaluate different non-invasive AVF flow measurements for detecting AVF stenosis. Twenty-three haemodialysis patients with suspected AVF stenosis are enrolled based on abnormal physical signs or high venous pressure during dialysis. Ultrasound dilution, urea dilution, Doppler ultrasonography, and fistulography are performed on all patients. The accuracy of three non-invasive methods is compared. Fistulography reveals AVF stenosis in 18 patients, 12 of whom have severe stenosis (greater than 50% stenosis in diameter). Concerning the location of the stenosis lesions, eight are at the inflow site, six at the outflow site, and four on both sites. Receiver operating characteristic curve analysis shows that Doppler ultrasonography has a high discriminative ability and the averaged areas under the curves are 0.933 (95% confidence interval [CI]; 0.81 to 0.99) for stenosis and 0.929 (95% CI 0.82-0.99) for severe stenosis. The sensitivity of each method for the prediction of access stenosis using ultrasound dilution, urea dilution, and Doppler ultrasonography is 73%, 73%, and 80%, respectively. The respective specificity of each method is 40%, 80%, and 100%, respectively. Physical examination (PE) shows an 80% sensitivity and 80% specificity in the detection of AVF stenosis. The combination of Doppler ultrasound with PE produces the highest sensitivity (93%) for detecting AVF stenosis. Doppler ultrasound combined with physical examination is more accurate than other non-invasive methods for detecting AVF stenosis.


Mifepristone and misoprostol for medical termination of pregnancy: the effectiveness of a flexible regimen.
Mifepristone, followed 48 hours later by administration of misoprostol, is a well-established regimen for medical termination of pregnancy (TOP). Although this regimen is effective, its inflexibility may limit its provision in an outpatient service. To confirm that misoprostol administration is effective whether administered 24, 48 or 72 hours after oral mifepristone. Observational study of 234 consecutive women with pregnancies up to 83 days' gestational age in whom medical TOP was performed during the period December 2000-July 2001. Women's Health Care Department, Royal Bolton Hospital, Bolton, UK. There was a high success rate for complete abortion in all groups whether mifepristone was administered 24, 48 or 72 hours prior to misoprostol. This study suggests that a more flexible regimen of mifepristone/misoprostol administration for medical TOP is effective in routine clinical practice.


Discrepancies between vertebral bone density values: the least dense vertebra.
Vertebral bone mineral density (BMD) measurements by DXA are considered reliable indicators of local fracture risk in the absence of radiographic deformities. The clinical evaluation of one individual vertebra presenting a BMD value significantly less than the others is attempted in this study. For a period of 30 months, BMD measurements of L1-L4 vertebrae and femoral neck (FN) were performed by DXA in 817 postmenopausal women, aged under 65 years, with a BMI less than 33 kg/m(2). In 204 (25%) of these women (group A), the least dense vertebra (LDV) presented a BMD value lower than 92.4% from the immediate denser vertebra. The remaining 613 women comprised group B. Women with X-ray proven vertebral degenerative lesions or deformities were excluded from the study. Among the four measured vertebrae, L1 was the most frequent LDV (47%), whilst L3 was the most rare (2%). Absolute and age-adjusted BMD values of L1-L4 and FN, as well as the proportions of osteopenic or osteoporotic women, did not differ significantly between the two groups. A significant positive correlation was observed between either L1-L4 or LDV and FN BMD values in both groups, but stepwise multiple regression analysis revealed that in group A the LDV did not participate in the model explaining the variability of the FN BMD values. In group B, the least dense vertebra was the only variable participating in the respective model (adjusted-R(2) = 37.7%). It is concluded that in a significant proportion of relatively young postmenopausal women, a wide variance of BMD values exists between individual vertebral BMD values without radiographic background. L1 was the most frequent LDV and L3 the most rare. In such cases, the evaluation of the least dense vertebra seems to offer an alternative estimation of vertebral bone mass, instead of mean L1-L4.


Humoral immune response to SARS-CoV-2 in five different groups of individuals at different environmental and professional risk of infection.
It is partially unknown whether the immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection persists with time. To address this issue, we detected the presence of SARS-CoV-2 antibodies in different groups of individuals previously diagnosed with COVID-19 disease (group 1 and 2), or potentially exposed to SARS-CoV-2 infection (group 3 and 4), and in a representative group of individuals with limited environmental exposure to the virus due to lockdown restrictions (group 5). The primary outcome was specific anti-SARS-CoV-2 antibodies in the different groups assessed by qualitative and quantitative analysis at baseline, 3 and 6 months follow-up. The seroconversion rate at baseline test was 95% in group 1, 61% in group 2, 40% in group 3, 17% in group 4 and 3% in group 5. Multivariate logistic regression analysis revealed male gender, close COVID-19 contact and presence of COVID-19 related symptoms strongly associated with serological positivity. The percentage of positive individuals as assessed by the qualitative and quantitative tests was superimposable. At the quantitative test, the median level of SARS-CoV-2 antibody levels measured in positive cases retested at 6-months increased significantly from baseline. The study indicates that assessing antibody response to SARS-CoV-2 through qualitative and quantitative testing is a reliable disease surveillance tool.


Comparison of filtration properties of hepatitis B virus, hepatitis C virus and simian virus 40 using a polyvinylidene fluoride membrane filter.
We examined the ability of a modified polyvinylidene fluoride membrane filter to remove blood-borne and surrogate viruses. Phages PR772 and PP7, hepatitis B virus (HBV), hepatitis C virus and simian virus 40 (SV40) were spiked in minimum essential media with 10% fetal calf serum and the concentration of these viruses compared before and after filtration by either plaque assay or polymerase chain reaction. Viruses >50 nm were removed to below detection limits (>10(6) logs) for all filters tested. A 5-log reduction of HBV (42 nm) and 2- to 3-log reduction of HCV (30-65 nm) was observed. A predictable size-based removal of viral agents was observed. The results also suggest the possible utility of SV40 as a surrogate to HBV for membrane filter challenge studies.


Levels of zinc, copper, cadmium, and lead in fruits and vegetables grown and consumed in Aseer Region, Saudi Arabia.
The levels of four metals (Zn, Cu, Cd, and Pb) were evaluated in two fruit types (apricot and fig), a fruity vegetable (tomato), and three leafy vegetables (arugula, spinach, and lettuce) that are commonly grown and consumed in Aseer Region, Saudi Arabia. Flame atomic absorption spectrophotometry was employed for quantification. The quality of results was checked by a certified reference material (NIST SRM 1570a). Good recovery values in the range of 87-104% were achieved. Metals were quantified in washed and unwashed samples to evaluate the effect of washing. Statistically, no significant difference was noticed (p>0.05), except for Zn in arugula and Cu in apricot and spinach. The levels of metals found in the analyzed fruits and vegetables were in their normal ranges in crops and not posing any serious risks to the consumers in Aseer Region. The toxic elements Pb and Cd were well below the maximum levels set in the Saudi and international food standards. Zn and Cu levels were comparable to the ranges reported in worldwide previous studies.


Effect of copper and zinc status on susceptibility to cadmium intoxication.
The effects of dietary cadmium on copper and zinc metabolism in animals are described. Emphasis is given to situations involving chronic exposure to low levels of cadmium, to the identification of population groups most at risk, and to the protective effect of dietary supplementation with copper and zinc. The mechanism of the interaction between the metals and the involvement of metallothionein are discussed.


Initial systolic blood pressure associates with systemic inflammation, myocardial injury, and outcomes in patients with acute coronary syndromes.
Outcomes after acute coronary syndromes (ACS) are determined by baseline risk profiles, including initial systolic blood pressure (sBP) levels. Herein, we aimed to characterize ACS patients stratified by initial sBP levels and study their relation to inflammation, myocardial injury and post-ACS outcomes. We analysed 4724 prospectively recruited ACS patients according to invasively assessed sBP (<100, 100-139, and ≥140 mmHg) at admission. Biomarkers of systemic inflammation [high-sensitivity C-reactive protein (hs-CRP)] and myocardial injury [high-sensitivity cardiac troponin T (hs-cTnT)] were measured centrally. Major adverse cardiovascular events (MACE; composite measure of non-fatal myocardial infarction, non-fatal stroke, and cardiovascular death) were externally adjudicated. Leukocyte counts, hs-CRP, hs-cTnT, and creatine kinase (CK) levels decreased from low to high sBP strata (ptrend < 0.001). Patients with sBP < 100 mmHg developed more often cardiogenic shock (CS; P < 0.001), and had a 1.7-fold increased multivariable-adjusted MACE risk at 30 days (HR 1.68, 95% CI 1.05-2.69, P = 0.031) which did not persist at one year (HR 1.38, 95% CI 0.92-2.05, P = 0.117). Those with sBP < 100 mmHg and CS showed a higher leukocyte count (P < 0.001), an increased neutrophil-to-lymphocyte-ratio (P = 0.031), and higher hs-cTnT and CK levels relative to those without CS (P < 0.001 and P = 0.002, respectively), whereas hs-CRP levels did not differ. Patients who developed CS had a 3.6- and 2.9-fold increased MACE risk at 30 days (HR 3.58, 95% CI 1.77-7.24, P < 0.001) and at one year (HR 2.94 95% CI, 1.57-5.53, P < 0.001), which was intriguingely attenuated after controlling for distinct inflammatory profiles. In patients with ACS, proxies of systemic inflammation and myocardial injury are inversely associated with initial sBP levels, with highest biomarker levels observed in those <100 mmHg. If linked to high levels of cellular inflammation, these patients are prone to develop CS and are at high MACE and mortality risk.


Phenotypic Screen with TSC-Deficient Neurons Reveals Heat-Shock Machinery as a Druggable Pathway for mTORC1 and Reduced Cilia.
Tuberous sclerosis complex (TSC) is a neurogenetic disorder that leads to elevated mechanistic targeting of rapamycin complex 1 (mTORC1) activity. Cilia can be affected by mTORC1 signaling, and ciliary deficits are associated with neurodevelopmental disorders. Here, we examine whether neuronal cilia are affected in TSC. We show that cortical tubers from TSC patients and mutant mouse brains have fewer cilia. Using high-content image-based assays, we demonstrate that mTORC1 activity inversely correlates with ciliation in TSC1/2-deficient neurons. To investigate the mechanistic relationship between mTORC1 and cilia, we perform a phenotypic screen for mTORC1 inhibitors with TSC1/2-deficient neurons. We identify inhibitors of the heat shock protein 90 (Hsp90) that suppress mTORC1 through regulation of phosphatidylinositol 3-kinase (PI3K)/Akt signaling. Pharmacological inhibition of Hsp90 rescues ciliation through downregulation of Hsp27. Our study uncovers the heat-shock machinery as a druggable signaling node to restore mTORC1 activity and cilia due to loss of TSC1/2, and it provides broadly applicable platforms for studying TSC-related neuronal dysfunction.


Intestinal expression of metal transporters in Wilson's disease.
In Wilson's disease (WND), biallelic ATP7B gene mutation is responsible for pathological copper accumulation in the liver, brain and other organs. It has been proposed that copper transporter 1 (CTR1) and the divalent metal transporter 1 (DMT1) translocate copper across the human intestinal epithelium, while Cu-ATPases: ATP7A and ATP7B serve as copper efflux pumps. In this study, we investigated the expression of CTR1, DMT1 and ATP7A in the intestines of both WND patients and healthy controls to examine whether any adaptive mechanisms to systemic copper overload function in the enterocytes. Duodenal biopsy samples were taken from 108 patients with Wilson's disease and from 90 controls. CTR1, DMT1, ATP7A and ATP7B expression was assessed by polymerase chain reaction and Western blot. Duodenal CTR1 mRNA and protein expression was decreased in WND patients in comparison to control subjects, while ATP7A mRNA and protein production was increased. The variable expression of copper transporters may serve as a defense mechanism against systemic copper overload resulting from functional impairment of ATP7B.


Association mapping through heuristic evolutionary history reconstruction-application to GAW15 Problem 3.
This paper presents a novel method of identifying phenotypically important regions of the genome. It involves a form of association mapping that works by summarizing properties of the ancestral recombination graph (ARG) of a sample of unrelated phenotyped and genotyped individuals. By breaking the sample into many small sub-samples and averaging the results, it becomes computationally tractable to measure the degree to which the evolutionary history of any locus is consistent with the distribution of the phenotypes in the sample. Analysis of simulated rheumatoid arthritis data demonstrates the efficiency and effectiveness of this method in identifying loci of large phenotypic effect.