Protein kinase C mediation of Ca(2+)-independent contractions of vascular smooth muscle.
Tumour-promoting phorbol esters induce slow, sustained contractions of vascular smooth muscle, suggesting that protein kinase C (PKC) may play a role in the regulation of smooth muscle contractility. In some cases, e.g., ferret aortic smooth muscle, phorbol ester induced contractions occur without a change in [Ca2+]i or myosin phosphorylation. Direct evidence for the involvement of PKC came from the use of single saponin-permeabilized ferret aortic cells. A constitutively active catalytic fragment of PKC induced a slow, sustained contraction similar to that triggered by phenylephrine. Both responses were abolished by a peptide inhibitor of PKC. Contractions of similar magnitude occurred even when the [Ca2+] was reduced to close to zero, implicating a Ca(2+)-independent isoenzyme of PKC. Of the two Ca(2+)-independent PKC isoenzymes, epsilon and zeta, identified in ferret aorta, PKC epsilon is more likely to mediate the contractile response because (i) PKC epsilon, but not PKC zeta, is responsive to phorbol esters; (ii) upon stimulation with phenylephrine, PKC epsilon translocates from the sarcoplasm to the sarcolemma, whereas PKC zeta, translocates from a perinuclear localization to the interior of the nucleus; and (iii) when added to permeabilized single cells of the ferret aorta at pCa 9, PKC epsilon, but not PKC zeta, induced a contractile response similar to that induced by phenylephrine. A possible substrate of PKC epsilon is the smooth muscle specific, thin filament associated protein, calponin. Calponin is phosphorylated in intact smooth muscle strips in response to carbachol, endothelin-1, phorbol esters, or okadaic acid. Phosphorylation of calponin in vitro by PKC (a mixture of alpha, beta, and gamma isoenzymes) dramatically reduces its affinity for F-actin and alleviates its inhibition of the cross-bridge cycling rate. Calponin is phosphorylated in vitro by PKC epsilon but is a very poor substrate of PKC zeta. A signal transduction pathway is proposed to explain Ca(2+)-independent contraction of ferret aorta whereby extracellular signals trigger diacylglycerol production without a Ca2+ transient. The consequent activation of PKC epsilon would result in calponin phosphorylation, its release from the thin filaments, and alleviation of inhibition of cross-bridge cycling. Slow, sustained contraction then results from a slow rate of cross-bridge cycling because of the basal level of myosin light chain phosphorylation (approximately 0.1 mol Pi/mol light chain). We also suggest that signal transduction through PKC epsilon is a component of contractile responses triggered by agonists that activate phosphoinositide turnover; this may explain why smooth muscles often develop more force in response, e.g., to alpha 1-adrenergic agonists than to K+.


Overexpression of mitochondrial sirtuins alters glycolysis and mitochondrial function in HEK293 cells.
SIRT3, SIRT4, and SIRT5 are mitochondrial deacylases that impact multiple facets of energy metabolism and mitochondrial function. SIRT3 activates several mitochondrial enzymes, SIRT4 represses its targets, and SIRT5 has been shown to both activate and repress mitochondrial enzymes. To gain insight into the relative effects of the mitochondrial sirtuins in governing mitochondrial energy metabolism, SIRT3, SIRT4, and SIRT5 overexpressing HEK293 cells were directly compared. When grown under standard cell culture conditions (25 mM glucose) all three sirtuins induced increases in mitochondrial respiration, glycolysis, and glucose oxidation, but with no change in growth rate or in steady-state ATP concentration. Increased proton leak, as evidenced by oxygen consumption in the presence of oligomycin, appeared to explain much of the increase in basal oxygen utilization. Growth in 5 mM glucose normalized the elevations in basal oxygen consumption, proton leak, and glycolysis in all sirtuin over-expressing cells. While the above effects were common to all three mitochondrial sirtuins, some differences between the SIRT3, SIRT4, and SIRT5 expressing cells were noted. Only SIRT3 overexpression affected fatty acid metabolism, and only SIRT4 overexpression altered superoxide levels and mitochondrial membrane potential. We conclude that all three mitochondrial sirtuins can promote increased mitochondrial respiration and cellular metabolism. SIRT3, SIRT4, and SIRT5 appear to respond to excess glucose by inducing a coordinated increase of glycolysis and respiration, with the excess energy dissipated via proton leak.


Large-scale transcriptome analysis of retroelements in the migratory locust, Locusta migratoria.
Retroelements can successfully colonize eukaryotic genome through RNA-mediated transposition, and are considered to be some of the major mediators of genome size. The migratory locust Locusta migratoria is an insect with a large genome size, and its genome is probably subject to the proliferation of retroelements. An analysis of deep-sequencing transcriptome data will elucidate the structure, diversity and expression characteristics of retroelements. We performed a de novo assembly from deep sequencing RNA-seq data and identified 105 retroelements in the locust transcriptome. Phylogenetic analysis of reverse transcriptase sequences revealed 1 copia, 1 BEL, 8 gypsy and 23 non-long terminal repeat (LTR) retroelements in the locust transcriptome. A novel approach was developed to identify full-length LTR retroelements. A total of 5 full-length LTR retroelements and 2 full-length non-LTR retroelements that contained complete structures for retrotransposition were identified. Structural analysis indicated that all these retroelements may have been activated or deprived of retrotransposition activities very recently. Expression profiling analysis revealed that the retroelements exhibited a unique expression pattern at the egg stage and showed differential expression profiles between the solitarious and gregarious phases at the fifth instar and adult stage. We hereby present the first de novo transcriptome analysis of retroelements in a species whose genome is not available. This work contributes to a comprehensive understanding of the landscape of retroelements in the locust transcriptome. More importantly, the results reveal that non-LTR retroelements are abundant and diverse in the locust transcriptome.


Maternal Genetic Ancestry and Legacy of 10(th) Century AD Hungarians.
The ancient Hungarians originated from the Ural region in today's central Russia and migrated across the Eastern European steppe, according to historical sources. The Hungarians conquered the Carpathian Basin 895-907 AD, and admixed with the indigenous communities. Here we present mitochondrial DNA results from three datasets: one from the Avar period (7(th)-9(th) centuries) of the Carpathian Basin (n = 31); one from the Hungarian conquest-period (n = 76); and a completion of the published 10(th)-12(th) century Hungarian-Slavic contact zone dataset by four samples. We compare these mitochondrial DNA hypervariable segment sequences and haplogroup results with published ancient and modern Eurasian data. Whereas the analyzed Avars represents a certain group of the Avar society that shows East and South European genetic characteristics, the Hungarian conquerors' maternal gene pool is a mixture of West Eurasian and Central and North Eurasian elements. Comprehensively analyzing the results, both the linguistically recorded Finno-Ugric roots and historically documented Turkic and Central Asian influxes had possible genetic imprints in the conquerors' genetic composition. Our data allows a complex series of historic and population genetic events before the formation of the medieval population of the Carpathian Basin, and the maternal genetic continuity between 10(th)-12(th) century and modern Hungarians.


Castleman's disease associated with calcifying fibrous tumor: A rare association with review of literature.
Castleman's disease is a rare disease characterized by unicentric or multicentric enlargement of the lymph node, and it is mostly of the hyaline vascular type. Calcifying fibrous tumor is a neoplasm characterized by large areas of hyalinized collagen with paucicellular areas along with dystrophic calcification and scattered lymphoplasmacytic infiltrates. There are only five case reports to describe an association between these two entities. We report a case of a 38-year-old male with seropositivity for hepatitis B virus presenting with a right-sided cervical swelling. Fine-needle aspiration cytology (FNAC) was done, and a diagnosis of follicular hyperplasia was offered. Biopsy confirmed the diagnosis of Castleman's disease. However, there was associated calcifying fibrous tumor in the lymph node induced by trauma due to FNAC.


Self-consistent field theory of polymer-ionic molecule complexation.
A self-consistent field theory is developed for polymers that are capable of binding small ionic molecules (adsorbates). The polymer-ionic molecule association is described by Ising-like binding variables, C(i) ((a))(kDelta)(=0 or 1), whose average determines the number of adsorbed molecules, n(BI). Polymer gelation can occur through polymer-ionic molecule complexation in our model. For polymer-polymer cross-links through the ionic molecules, three types of solutions for n(BI) are obtained, depending on the equilibrium constant of single-ion binding. Spinodal lines calculated from the mean-field free energy exhibit closed-loop regions where the homogeneous phase becomes unstable. This phase instability is driven by the excluded-volume interaction due to the single occupancy of ion-binding sites on the polymers. Moreover, sol-gel transitions are examined using a critical degree of conversion. A gel phase is induced when the concentration of adsorbates is increased. At a higher concentration of the adsorbates, however, a re-entrance from a gel phase into a sol phase arises from the correlation between unoccupied and occupied ion-binding sites. The theory is applied to a model system, poly(vinyl alcohol) and borate ion in aqueous solution with sodium chloride. Good agreement between theory and experiment is obtained.


Tissue engineering of flexor tendons: the effect of a tissue bioreactor on adipoderived stem cell-seeded and fibroblast-seeded tendon constructs.
Tissue-engineered flexor tendons could eventually be used for reconstruction of large tendon defects. The goal of this project was to examine the effect of a tissue bioreactor on the biomechanical properties of tendon constructs seeded with adipoderived stem cells (ASCs) and fibroblasts (Fs). Rabbit rear paw flexor tendons were acellularized and seeded with ASCs or Fs. A custom bioreactor applied a cyclic mechanical load of 1.25 N at 1 cycle/minute for 5 days onto the tendon constructs. Three additional groups were used as controls: fresh tendons and tendons reseeded with either ASCs or Fs that were not exposed to the bioreactor treatment and were left in stationary incubation for 5 days. We compared the ultimate tensile stress (UTS) and elastic modulus (EM) of bioreactor-treated tendons with the unloaded control tendons and fresh tendons. Comparison across groups was assessed using one-way analysis of variance with the significance level set at p<.05. Pairwise comparison between the samples was determined by using the Tukey test. The UTS and EM values of bioreactor-treated tendons that were exposed to cyclic load were significantly higher than those of unloaded control tendons. Acellularized tendon constructs that were reseeded with ASCs and exposed to a cyclic load had a UTS of 66.76 MPa and an EM of 906.68 MPa; their unloaded equivalents had a UTS of 47.90 MPa and an EM of 715.57 MPa. Similar trends were found in the fibroblast-seeded tendon constructs that were exposed to the bioreactor treatment. The bioreactor-treated tendons approached the UTS and EM values of fresh tendons. Histologically, we found that cells reoriented themselves parallel to the direction of strain in response to cyclic strain. The application of cyclic strain on seeded tendon constructs that were treated with the bioreactor helped achieve a UTS and an EM comparable with those of fresh tendons. Bioreactor pretreatment and alternative cell lines, such as ASCs and Fs, might therefore contribute to the in vitro production of strong tendon material.


Multiloculated Cystic Type Renal Epithelioid Angiomyolipoma Mimicking Renal Cell Carcinoma: A Case Report.
Renal epithelioid angiomyolipoma (EAML) is a rare variant of angiomyolipoma (AML), with a prominent epithelioid component. EAML usually presents as a large heterogeneous soft tissue lesion with intratumoral hemorrhage and variable necrosis or cystic changes. We present a case of multiloculated cystic renal EAML mimicking renal cell carcinoma in a 64-year-old female. Intracystic massive hemorrhage, hyperattenuating wall and septa on an unenhanced study, and enlarged intratumoral vessels can be helpful imaging features for distinguishing renal EAML from renal cell carcinoma.


Symptomatic gastroesophageal reflux as a risk factor for esophageal adenocarcinoma.
The causes of adenocarcinomas of the esophagus and gastric cardia are poorly understood. We conducted an epidemiologic investigation of the possible association between gastroesophageal reflux and these tumors. We performed a nationwide, population-based, case-control study in Sweden. Case ascertainment was rapid, and all cases were classified uniformly. Information on the subjects' history of gastroesophageal reflux was collected in personal interviews. The odds ratios were calculated by logistic regression, with multivariate adjustment for potentially confounding variables. Of the patients interviewed, the 189 with esophageal adenocarcinoma and the 262 with adenocarcinoma of the cardia constituted 85 percent of the 529 patients in Sweden who were eligible for the study during the period from 1995 through 1997. For comparison, we interviewed 820 control subjects from the general population and 167 patients with esophageal squamous-cell carcinoma. Among persons with recurrent symptoms of reflux, as compared with persons without such symptoms, the odds ratios were 7.7 (95 percent confidence interval, 5.3 to 11.4) for esophageal adenocarcinoma and 2.0 (95 percent confidence interval, 1.4 to 2.9) for adenocarcinoma of the cardia. The more frequent, more severe, and longer-lasting the symptoms of reflux, the greater the risk. Among persons with long-standing and severe symptoms of reflux, the odds ratios were 43.5 (95 percent confidence interval, 18.3 to 103.5) for esophageal adenocarcinoma and 4.4 (95 percent confidence interval, 1.7 to 11.0) for adenocarcinoma of the cardia. The risk of esophageal squamous-cell carcinoma was not associated with reflux (odds ratio, 1.1; 95 percent confidence interval, 0.7 to 1.9). There is a strong and probably causal relation between gastroesophageal reflux and esophageal adenocarcinoma. The relation between reflux and adenocarcinoma of the gastric cardia is relatively weak.


Respiratory paralysis during treatment of hypertension with trimethaphan camsylate.
Four cases are reported in which respiratory arrest occured coincident with the intravenous administration of large doses of trimethaphan camsylate (Arfonad) to control hypertension. The mechanism of the respiratory depression is unknown, but it may have been related to a direct effect of trimethaphan on the respiratory center or to a curare-like effect of the drug. Close monitoring of ventilatory capacity should be maintained in all patients treated with trimethaphan.


Severe extrapiramidal symptoms after nonintentional risperidone exposure in a child: case report and review of the literature.
Increase in use of atypical antipsychotics has been paralleled by an increase in the incidence of intentional and nonintentional overdose. Pediatric cases are uncommon, but may be severe. We describe a case of a child presenting with severe extrapiramidal symptoms, initially interpreted as seizures, caused by a nonintentional intoxication with risperidone, and review management options and the literature.


Behavioural motor effects of MK-801 and DNQX parenteral administration in adult cats: dose-response analysis. Modulatory role of dopaminergic D1 and D2 antagonists on MK-801 induced motor behaviours.
1. Administration of MK-801 a selective antagonist of the NMDA receptors (50, 100 and 150 micrograms/kg, s.c.) elicited in adult cats ataxia and loss of equilibrium. A dose-response effect was observed. 2. Administration of DNQX, a selective antagonist of the non-NMDA receptors, even with doses 20 times higher than those employed with MK-801, did not produce any behavioural disturbances. 3. Previous injection of SCH 23390, a selective parenteral antagonist of dopamine D1 receptor, reduced significantly the intense ataxic effects of MK-801, while sulpiride only increased the latency of the symptoms. 4. The results are discussed considering the reported interactions between the dopaminergic and glutamatergic systems.


Enteral glutamate is the preferential source for mucosal glutathione synthesis in fed piglets.
To measure the source and rate of mucosal glutathione (GSH) synthesis, fed piglets (28 days old; 7.7 kg) received a 6-h infusion of intragastric [U-13C]glutamate (n = 11) either with (n = 5) or without (n = 6) an intragastric infusion of [1-13C]glycine (0-6 h) and [1,2-13C2(U-13C)]glycine (3-6 h). Eighty-four percent of the labeled mucosal GSH-glutamate and 86% of the luminal GSH-glutamate was 13C5. The tracer-to-tracee ratio of GSH-[U-13C]glutamate was 75% of that of mucosal glutamate. Sixty percent of the labeled mucosal glutamate was 13C1, 13C2, or 13C3, but the tracer-to-tracee ratios of these isotopomers in GSH-glutamate were not significantly different from zero. After 3 h of infusion, the tracer-to-tracee ratio of GSH-[U-13C]glycine was 46%, and after 6 h of infusion GSH-[13C1]glycine was 82% of that of mucosal glycine. This suggested that the half-life of mucosal GSH was 2.7 +/- 0.1 h. We concluded that, in fed piglets, mucosal GSH-glutamate derived largely from the direct metabolism of enteral glutamate rather than from glutamate that was metabolized within the mucosa.


Attendance at National Cancer Institute and Children's Oncology Group Facilities for Children, Adolescents, and Young Adults with Cancer in Pennsylvania: A Population-Based Study.
Purpose: Adolescents and young adults (AYAs) with cancer are a vulnerable population with decreased attendance at National Cancer Institute (NCI) comprehensive cancer centers and Children's Oncology Group (COG) facilities. Decreased attendance at NCI/COG facilities has been associated with poor cancer outcomes. The objective of this study was to evaluate cancer care patterns of AYAs compared with children, within Pennsylvania, and factors associated with attending an NCI/COG facility. Methods: Data from the Pennsylvania Cancer Registry between 2010 and 2015 for patients aged 0-39 years at cancer diagnosis were used. Primary analyses focused on age at diagnosis, insurance status, race, ethnicity, gender, cancer type, stage, diagnosis year, and distance to the NCI/COG facility. The primary outcome was receipt of care at an NCI/COG facility. Odds ratios (ORs) were calculated using multivariable logistic regression models. Sensitivity analyses were conducted to test and estimate robustness. Results: A sample of 15,002 patients, ages 0-39, was obtained, including 8857 patients (59%) who attended an NCI/COG facility. Patients were significantly less likely to attend an NCI/COG facility if they were aged 31-39 years (OR 0.054, 95% confidence interval [CI] 0.04-0.07), non-White (OR 0.890, 95% CI 0.80-0.99), Hispanic (OR 0.701, 95% CI 0.59-0.83), female (OR 0.915, 95% CI 0.84-1.00), had Medicaid insurance (OR 0.836, 95% CI 0.75-0.93), and lived further from an NCI/COG facility. Sensitivity analyses largely corroborated the performed estimates. Conclusions: AYAs with cancer in Pennsylvania have disproportionate attendance at specialized NCI/COG facilities across a variety of demographic domains. Enhancing the attendance of AYAs with cancer at these specialized centers is crucial to improve cancer outcomes.


[Possibilities for automatic quantitative image analysis of cell organelles stained ultrahistochemically].
Metal-containing secretory granules of alpha 2-cells of the Langerhans' islets of normal and alloxandiabetic rats were stained for electron microscopy by a special method. Thereafter the electron micrographs were analysed by the Quantimet 720 image analysing computer with respect to size of the granules and their quantitative distribution in classes. The following statistical tests verified significant differences between the distributions of the granules under normal and diabetic conditions. The secretory granules from alloxandiabetic animals were diminished distinctly. The examination by image analysing computer confirmed objectively observations, we have made and published formerly.


Clinical and diagnostic tests for shoulder disorders: a critical review.
The shoulder is one of the most complex joints in the human body and, as such, presents an evaluation and diagnostic challenge. The first steps in its evaluation are obtaining an accurate history and physical examination and evaluating conventional radiography. The use of other imaging modalities (eg, ultrasound, magnetic resonance imaging and computed tomography) should be based on the type of additional information needed. The goals of this study were to review the current limitations of evidence-based medicine with regard to shoulder examination and to assess the rationale for and against the use of diagnostic physical examination tests.


Control of abdominal extension in the freely moving intact crayfish cherax destructor. I. Activity Of the tonic stretch receptor
Electrical recordings were made from the sensory neurone of the tonic muscle receptor organ in the abdomen of the intact, freely behaving crayfish Cherax destructor. Slow extensions of the abdomen were evoked by lowering a platform from beneath the suspended crayfish, and the movements and tonic sensory neurone activity were video-recorded simultaneously. The recordings showed that the tonic sensory neurone was active when the abdomen was fully flexed prior to the extension. When the extension began, however, the sensory neurone ceased firing shortly after movement was detected, irrespective of the load applied to the abdomen. When the abdomen was physically blocked from extending fully, the sensory neurone did not fire. The tonic muscle receptor organ is considered to be the length-detecting sensor for a load-compensating servo-loop, but the results demonstrate that its activity pattern during extensions evoked by a platform-drop in C. destructor are not consistent with that role.


Postoperative cognitive dysfunction after noncardiac surgery: a systematic review.
This article describes a systematic review on the research into postoperative cognitive dysfunction (POCD) in noncardiac surgery to ascertain the status of the evidence and to examine the methodologies used in studies. The review demonstrated that in the early weeks after major noncardiac surgery, a significant proportion of people show POCD, with the elderly being more at risk. Minimal evidence was found that patients continue to show POCD up to 6 months and beyond. Studies on regional versus general anesthesia have not found differences in POCD. Many studies were found to be underpowered, and a number of other methodologic difficulties were identified. These include the different types of surgery in studies and variations in the number and range of neuropsychological tests used. A particular issue is the variety of definitions used to classify individuals as having POCD.


Hypnotherapy of a pain disorder: a clinical case study.
Hypnotherapy's effectiveness in improving and controlling chronic pain of various etiologies has been demonstrated by studies; the mechanism by which hypnosis does this is more complex than a simple induction of muscle relaxation. This study reveals, in addition to this mechanism, a deeper dimension of hypnotherapy from the vantage of a patient with a medical-surgical background, diagnosed with a pain disorder and major severe depressive disorder in addition to incurable painful symptoms, through treatment associated with hypnoanalysis. Following psychotherapy, which included some elements of cognitive-behavioral therapy, a complete remission of the anxious-depressive mood and the painful symptoms was achieved.


Genomic imbalances in key ion channel genes and telomere shortening in sudden cardiac death victims.
Sudden cardiac death (SCD) can be caused by a number of reasons. Previous works have identified the genetic causes, such as alterations in the DNA sequence, for many of these diseases. We hypothesize that some patients may show genomic imbalances and changes in the gene copy number leading to genetic instability. To clarify this, we analysed DNA samples from SCD victims using comparative genomic hybridization (CGH), a molecular cytogenetic technique that permits the genome-wide screening of chromosomal imbalances, and telomere length measurement. DNA derived from peripheral blood and heart tissue of 14 SCD cases and six apparently healthy control individuals were subjected to CGH analysis. Telomere length measurements were done by the Southern blotting method. Eight out of 14 SCD cases exhibited changes in DNA/gene copy number. CGH analysis showed variation in the gene copy number of some of the genes associated with potassium (KCNAB1, KCNH2, and KCNA4) and calcium (RyR2, ATP2A2) ions which are involved in maintaining the ionic balance of the heart. Alterations in TERC and TERT genes were also detected in SCD victims. In nine SCD victims shorter telomeres were detected. This might have resulted from excessive cellular proliferation and/or oxidative stress in these individuals. Copy number changes observed and telomere shortening detected in SCD cases would possibly explain at least some of the causes of SCD at early ages in humans. Identification of biomarkers of SCD is of great importance and thus the present study will facilitate the identification of some of the biomarkers.


Identification of twin pairs from large population-based samples.
The basis of most twin studies is the ascertainment of twins, often through twin registries, and determination of zygosity. The current rate of twin births in many industrialized countries implies that in the near future around 3% or more of individuals will be a twin. Hence, there are and will be a lot of twins around and many of those will not participate in twin studies. However, if large population-based samples are available that include appropriate identifiers, then twins can be detected and twin studies performed, even in the absence of zygosity information. We quantified the number of twin pairs that could be detected from a longitudinal survey in the Netherlands, which aims to answer questions about educational strategies and performance in primary education in the Netherlands. We detected 2865 twin pairs if we used a coded name identifier, date of birth, school, grade and year of survey, which is 2.01% of 284,945 pupils in five cohorts. Relaxing our selection criteria increased the number of apparent twin pairs identified, most of which are false positives due to chance matching of identification criteria. We show that the intraclass correlation on measured phenotypes can be used as a quality control measure for twin identification, and quantify the proportion of false negatives (true twin pairs not identified) due to missing data and data coding errors. We compared our estimated rate of twins in the sample to census data and estimate that with our most stringent selection criteria we detect more than 80% of all twin pairs in the sample. We conclude that the identification of twin pairs from large population-based samples is feasible, rapid and accurate if the appropriate identifiers are available, and that twin pairs from such sources are a valuable resource for studies to answer scientific question about twins versus nontwins and about genetic and environmental factors of twin resemblance.


Results of combined surgery by phacoemulsification and vitrectomy.
Eyes scheduled for posterior segment surgery may have cataract, which obscures the visualization of the retina. Surgery may be carried out either by a two-step procedure: i.e., removal of the cataract followed later by posterior segment surgery; or it may be done in a single session: i.e., combined surgery of both the anterior and posterior segments. To evaluate the outcomes of combined surgery by phacoemulsification and vitrectomy. We retrospectively reviewed the records of 42 patients with coexisting cataract and vitreoretinal disease who underwent combined surgery by phacoemulsification and pars plana vitrectomy at one session. Indications for surgery were vitreous hemorrhage in 71.4%, retinal detachment in 11.9%, macular hole in 11.9%, and epiretinal membrane in 4.8%. There were no significant intraoperative complications. The main early postsurgical complications were fibrinous formation in 11.9%, elevated intraocular pressure in 23.8%, and recurrent vitreous hemorrhage in 9.5%. There were a few late complications related to phacoemulsification: posterior synechia in 9.5%, posterior capsular opacification in 7.1%, and dislocating intraocular lens in 4.8%. Recurrent retinal detachment occurred in five eyes and rubeoisis iridis in one. Visual acuity was improved in 85.8%, stable in 7.1% and worse in 7.1%. Phacoemulsification performed at the time of posterior segment surgery enables good visualization during the vitrectomy, facilitates surgery, and is associated with only minor complications. In cases with cataract and vitreoretinal diseases, combined surgery by phacoemulsification and vitrectomy in one session may be considered.


Effects of therapeutic touch on anxiety, vital signs, and cardiac dysrhythmia in a sample of Iranian women undergoing cardiac catheterization: a quasi-experimental study.
To investigate the effects of Therapeutic Touch (TT) on anxiety, vital signs, and cardiac dysrhythmia in women undergoing cardiac catheterization. It was a quasi-experimental study. The participants had no history of hallucination, anxiety, or other psychological problems. Participants had to be conscious and have attained at least sixth-grade literacy level. Participants were randomly assigned into an intervention group (n = 23; received 10-15 minutes TT), a placebo group (n = 23; received 10-15 minutes simulated touch), and a control group (n = 23; did not receive any therapy). Data were collected using Spielberger's anxiety test, cardiac dysrhythmia checklist, and vital signs recording sheet. Statistical analyses were considered to be significant at α = .05 levels. Sixty-nine women ranging in age from 35 to 65 years participated. TT significantly decreased state anxiety p < 0.0001 but not trait anxiety (p = .88), decreased the incidence of all cardiac dysrhythmias p < 0.0001 except premature ventricular contraction (p = .01), and regulated vital signs p < 0.0001 in the intervention group versus placebo and control group. TT is an effective approach for managing state anxiety, regulating vital signs, and decreasing the incidence of cardiac dysrhythmia during stressful situations, such as cardiac catheterization, in Iranian cardiac patients.


Nimbus-6 earth radiation budget experiment.
This paper describes the Nimbus-6 earth radiation budget experiment including its prelaunch calibration and in-flight performance. A preliminary assessment of the data shows the ERB measurement of the solar constant to be 1392 W/m(2) which is 1.6% higher than the expected value of 1370 W/m(2). Both values are traceable to the cavity radiometer scale. There is a disagreement between the fixed wide-angle and scanning narrow-angle measurements of planetary outgoing longwave radiation flux. Since the scanning channels are calibrated in-flight and show good agreement with previous observations of the Nimbus-3 satellite, the discrepancy is believed to be due to erroneous wide-angle flux estimates. The erroneous estimates may be caused by the misinterpretation of the transfer function for the wide-angle-earth-flux sensing thermopile detectors when viewing the earth which, unlike the prelaunch calibration source, does not fill the field of view of the detector and is not an isotropic radiation source. A field of view factor for the wide-angle channels is determined using an in-flight calibration procedure using the night-time scanning channel longwave radiation flux measurements as the absolute standard. The planetary global albedoes, longwave radiation fluxes, and net radiation are about 30%, 240 W/m(2), and -4 W/m(2) for the months of July and August 1975, which is in good agreement with previous Nimbus-3 estimates.


Recognition of stimulus orientation.
The threshold for detecting a deviation in the orientation of a test stimulus from vertical and from horizontal was measured with a single test interval. The test stimuli were step edges, thin lines, and sine wave gratings of 2.5, 5.0 and 10.0 c/deg. Recognition thresholds were found to be independent of the frequency content and nature of the test stimulus. The average threshold value on the principal meridians was found to be approx. 0.6 deg. Additional testing was conducted with two interval forced choice designs, both with and without random perturbation of the orientation of the stimuli. Random perturbation of orientation reduces acuity estimates. The results are compared with those obtained by testing on the oblique meridia. We conclude that the meridional anisotropy of orientation discrimination, which favours the vertical and horizontal is a result, in part, of the influence of non-visual mechanisms.


Impaired energy utilization and Na-K-ATPase in diabetic peripheral nerve.
Recent electrophysiological and biochemical evidence implicates altered peripheral nerve Na-K-ATPase activity in the nerve conduction impairment of acute experimental diabetes. Composite in vitro nerve energy utilization is seriously impaired by experimental diabetes, yet is not modulated directly by insulin action on peripheral nerve. Therefore, we hypothesized that the reduction in diabetic nerve energy utilization reflects impaired nerve Na-K-ATPase activity. The reduction in steady-state energy utilization in diabetic peripheral nerve is shown to be quantitatively equal to the ouabain-inhibitable fraction of respiration, a measure of Na-K-ATPase activity in peripheral nerve. Na-K-ATPase activity in diabetic (but not nondiabetic) endoneurial preparations is influenced by medium solute concentration. Furthermore, diabetic nerve Na-K-ATPase activity and sodium-dependent myo-inositol uptake are similarly affected by medium solute changes, suggesting that the nerve sodium gradient may limit intracellular myo-inositol uptake in diabetic nerve. Conversely, because reduced diabetic nerve myo-inositol content impairs nerve Na-K-ATPase, a possible pathophysiological cycle of progressively deranged myo-inositol metabolism and Na-K-ATPase function may exist in diabetic peripheral nerve.


Flow cytometric analysis of the effect of sodium chloride on gastric cancer risk in the rat.
Dietary sodium chloride has been identified, both experimentally and epidemiologically, as a risk factor for gastric cancer. In order to elucidate the manner in which salt increases gastric tumor incidence in N-methyl-N'-nitro-N-nitrosoguanidine-treated animals, flow cytometric cell cycle analyses were performed on rats which had been treated with 1 ml of a solution of saturated NaCl by gavage and sacrificed 0, 1, 6, 12, 24, or 48 h after treatment. The gastric antra were excised, disaggregated, and stained with propidium iodide for cell cycle analysis. Results showed that there is a reduction in cell yield at early time points due to the toxicity of NaCl, followed by a net increase in the number of cells in the S phase of the cell cycle at 24 h. Treatment of rats with NaCl 24 h prior to a dose of 10 micrograms of 3H-labeled N-methyl-N'-nitro-N-nitrosoguanidine did not lead to an increase in alkylation of DNA isolated from mucosal cells. Therefore, the hypothesis that salt enhances gastric cancer risk from N-methyl-N'-nitro-N-nitrosoguanidine by disruption of the "mucosal barrier" leading to an increased effective dose to target cells is not supported by the results of these experiments. Several studies have shown that cells in S phase are the most susceptible to mutagenesis and that increasing the number of cycling cells in a target organ will increase tumor incidence (e.g., partial hepatectomy). Thus it is possible that NaCl increases gastric cancer risk through the mitogenesis which results from the damage caused to the mucosa by this agent.


The HR motif in the RNA-dependent RNA polymerase L protein of Chandipura virus is required for unconventional mRNA-capping activity.
Chandipura virus (CHPV) is an emerging human pathogen associated with acute encephalitis and is related closely to vesicular stomatitis virus (VSV), a prototype rhabdovirus. Here, we demonstrate that the RNA polymerase L protein of CHPV exhibits a VSV-like RNA:GDP polyribonucleotidyltransferase (PRNTase) activity, which transfers the 5'-monophosphorylated (p-) viral mRNA start sequence to GDP to produce a capped RNA, and that the conserved HR motif in the CHPV L protein is essential for the PRNTase activity. Interestingly, the CHPV L protein was found to form two distinct SDS-resistant complexes with the CHPV mRNA and leader RNA start sequences; mutations in the HR motif significantly reduced the formation of the former complex (a putative covalent enzyme-pRNA intermediate in the PRNTase reaction), but not the latter complex. These results suggest that the rhabdoviral L proteins universally use the active-site HR motif for the PRNTase reaction at the step of the enzyme-pRNA intermediate formation.


Epidemiology of Vasculitides in Khorasan Province, Iran.
Vasculitides are a heterogeneous group of more than 20 diseases defined by inflammation and destruction of blood vessels. We aimed to study the demographic characteristics of the primary vasculitides in the North East of Iran. We retrospectively studied the medical records of patients diagnosed with any kind of vasculitis at the Clinic and Department of Rheumatology of the Imam Reza Hospital, Mashhad, Iran between January 1, 2002, and December 31, 2012. Patients were classified according to the American College of Rheumatology 1990 criteria for the classification of vasculitis and the 2012 Revised International Chapel Hill Consensus Conference Nomenclature of Vasculitides. A total of 721 patients (51.5% male, 48.5% female) with a diagnosis of primary vasculitis was identified. The frequency distributions of vasculitic disorders were as follows: Behcet's disease, 63.6%; cutaneous leukocytoclastic angiitis, 8.2%; granulomatosis with polyangiitis (Wegener's), 6.8%; Takayasu's arteritis. 6%; giant cell arteritis, 4%; polyarteritis nodosa, 2.1%; microscopic polyangiitis, 0.6%; eosinophilic granulomatosis with polyangiitis (Churg-Strauss), 1.8%; cryoglobulinemic vasculitis, 0.3%; and IgA vasculitis (Henoch-Schonlein purpura), 3.5%. In our population, the most common forms of vasculitis are Behcet's disease, cutaneous leukocytoclastic angiitis, and granulomatosis with polyangiitis (Wegener's).


Ministry of Health computerisation programme.
The purpose of the paper is to give the readers an idea of the state of computerisation in the Ministry of Health, Singapore and to highlight some of the benefits of computerisation. The Ministry employs a wide range of computer systems from portable microcomputers, point-of-sale microcomputers, supermicros, minicomputers to mainframe computer. What are the roles of these computers? Why and how are they interconnected? Carry on reading if these questions are appealing to you.


Staring at the cold sun: blue light regulation is distributed within the genus Acinetobacter.
We previously showed that the opportunistic nosocomial pathogen Acinetobacter baumannii is able to sense and respond to light via BlsA, a BLUF (Blue-Light-sensing Using FAD)-domain photoreceptor protein. Here, we extend our previous studies showing that light regulation is not restricted to A. baumannii, but rather widespread within the genus Acinetobacter. First, we found that blue light modulates motility and biofilm formation in many species of the genus, including members of the Acinetobacter calcoaceticus-A. baumannii complex. In many of these species blue light acts as a key factor guiding the decision between motility or sessility at 24°C, whereas in A. baumannii, light inhibits both motility and biofilm formation. We also show that light regulation of motility occurred not only at 24°C but also at 37°C in non-A. baumannii species, contrasting the situation of A. baumannii which only shows photoregulation at 24°C. Second, we show that Acinetobacter baylyi (strain ADP1) BLUF-photoreceptors can functionally replace in vivo the A. baumannii 17978 BlsA protein and that the pathways leading to biofilm formation are inversely regulated at 24°C between these two microorganisms. Finally, we found the presence of predicted genes coding BLUF-containing proteins in all Acinetobacter sequenced genomes, even though the copy number is variable among them. Phylogenetic analysis suggests a common origin for all BLUF domains present in members of this genus, and could distinguish well-differentiated clusters that group together BLUF homologs from different species, a situation particularly clear for members of the ACB complex. Despite a role played by these BLUF domain-containing proteins in the photoregulation observed in the members of the genus Acinetobacter is a likely scenario given our findings in A. baumannii and A. baylyi, further research will contribute to confirm this possibility.


Surnames and Gender in Japan: Women's Challenges in Seeking Own Identity.
[[Japansurnamefamilygendermarriagelaw ]] Strongly influenced by the previous Meiji Civil Code that shaped people's perceptions about the traditional Japanese family, postwar Japanese society has not fully guaranteed gender equality, and whether to legally allow the dual-surname system is one of the major legal and political debates in Japanese society. To understand the tension between the traditional Japanese family emphasized in the previous Meiji Civil Code and gender equality emphasized in the current Japanese law, this study explored the surname system in Japan by reviewing historical trends, recent surveys, political debates, and comparing with other nations. This study illustrated that the surname was not attached to the family lineage and membership as today in the past, and symbolic significance of the surname has changed through the course of Japanese history.


Neutrophil surface presentation of the anti-neutrophil cytoplasmic antibody-antigen proteinase 3 depends on N-terminal processing.
The neutrophil serine protease proteinase 3 (PR3) is a main autoantigen in anti-neutrophil cytoplasmic antibody-associated vasculitis. PR3 surface presentation on neutrophilic granulocytes, the main effector cells, is pathogenically important. PR3 is presented by the NB1 (CD177) glycoprotein, but how the presentation develops during neutrophil differentiation is not known. An N-terminally unprocessed PR3 (proPR3) is produced early during neutrophil development and promotes myeloid cell differentiation. We therefore investigated if PR3 presentation depended on NB1 during neutrophil differentiation and if PR3 and proPR3 could both be presented by NB1. In contrast to mature neutrophils, differentiating neutrophils showed an early NB1-independent PR3 surface display that was recognized by only two of four monoclonal anti-PR3 antibodies and occurred in parallel with proPR3, but not PR3 secretion, suggesting that the NB1-independent surface PR3 was proPR3. PR3 gene expression preceeded NB1. When the NB1 receptor was detected on the surface, a mode of PR3 surface display similar to mature neutrophils developed together with the degranulation system. Ectopic expression studies showed that NB1 was a sufficient receptor for PR3 but not proPR3. ProPR3 display on the plasma membrane may influence the bone marrow microenvironment. NB1-mediated PR3 presentation depended on PR3 N-terminal processing implicating the PR3-N-terminus as NB1-binding site.


Correlation between intracellular cAMP content, kinematic parameters and hyperactivation of human spermatozoa after incubation with pentoxifylline.
Various compounds have been used in the attempt to improve sperm motility, including pentoxifylline (PF), a methylxanthine derivative. It has been postulated that PF, being a phosphodiesterase inhibitor, increases sperm kinematic parameters and the number of spermatozoa exhibiting hyperactivated motility by raising the intracellular content of cAMP, a molecule involved in the generation of sperm energy. However, it has not been clarified whether the biological effects of PF on sperm motility correlate with its ability to increase intracellular cAMP levels. To examine this relationship, the kinematic parameters, hyperactivation, and intracellular cAMP content were evaluated in motile spermatozoa, obtained by discontinuous Percoll gradient and swim-up from 21 normozoospermic semen samples, incubated without and with PF for 0, 1, 2, and 4 h. PF increased beat cross frequency after 1 and 2 h of incubation, curvilinear velocity and lateral head displacement (ALH) after 4 h, and hyperactivation after 1, 2, and 4 h, and decreased linearity (LIN) after 1 h of incubation. The intracellular cAMP content of spermatozoa incubated with PF increased at all time-points examined. Both intracellular cAMP content and increase in hyperactivation in response to PF decreased with the length of incubation. In the absence of PF, cAMP content was unchanged and was correlated significantly only with ALH and the percentage of spermatozoa with hyperactivated motility. Following incubation with PF, cAMP content correlated with hyperactivation and all sperm kinematic parameters, with the exception of LIN and straightness. These findings suggest that the beneficial effects of PF on sperm kinematic parameters and hyperactivation are related to its ability to increase intracellular cAMP content.


Dickkopf-Related Protein 2 is Epigenetically Inactivated and Suppresses Colorectal Cancer Growth and Tumor Metastasis by Antagonizing Wnt/β-Catenin Signaling.
Aberrant activation of the Wnt/β-catenin signaling pathway plays a key role in the pathogenesis of multiple tumors including digestive cancers. Recent studies have reported that Dickkopf-related protein 2 (DKK2) is epigenetically inactivated in numerous types of cancers and that its gene products exhibit tumor-suppressive properties. However, the biological functions and underlying molecular mechanisms of DKK2 in colon carcinoma remains obscure. We examined the expression of DKK2 in colon tumor cell lines by RT-PCR and its promoter methylation status in colon tumor cell lines and primary tumors by methylation-specific PCR (MSP). Ectopic expression of DKK2 was measured by RT-PCR prior to the other experiments. To investigate the function of DKK2, we assayed colony formation and cell proliferation, utilized flow cytometric analyses of the cell cycle and acridine orange/ethidium bromide (AO/EB) fluorescence staining for apoptosis, and examined wound healing, transwell migration and tumor growth in vivo. Western blots were used to explore the mechanisms of DKK2 in epithelial- mesenchymal transition and canonical Wnt/β-catenin signaling. We show here that downregulation or silencing of DKK2 was closely associated with the hypermethylation status of its promoter and that DKK2 expression could be restored by demethylation treatment. Methylation of the DKK2 promoter was detected in nearly all tumors and tumor-adjacent tissues, but not in normal colon tissues. Ectopic expression of DKK2 in colon cell lines HCT116 and HT-29 inhibited colony formation and cell viability by inducing cell cycle G0/G1 arrest and apoptosis, and growth of stable DKK2-infected HCT116 cells in nude mice was decreased compared to controls. Furthermore, DKK2 restrained cell migration through partial reversal of epithelial-to- mesenchymal transition and also by downregulating several stem cell markers. Our data further showed that restoration of DKK2 expression resulted in downregulation of active β-catenin and its downstream target genes. DKK2 appears to be a functional tumor suppressor regulating tumorigenesis of colorectal cancer by antagonizing Wnt/β-catenin signaling.


Testicular oxytocin: effects of intratesticular oxytocin in the rat.
The long-term effects of oxytocin administration on the testis were studied using intratesticular implants. Adult male rats had an Accurel device containing 20 micrograms oxytocin (releasing approximately 200 ng/day) implanted into the parenchyma of each testis; control animals received empty devices. The animals were killed at weekly intervals for 4 weeks. Some animals were perfused and the testes processed for light and electron microscopy. Blood was collected from the remaining animals for the measurement of testosterone, dihydrotestosterone, LH, FSH and oxytocin; epididymal sperm counts were measured and the testes were extracted and radioimmunoassayed for testosterone, dihydrotestosterone and oxytocin. Long-term administration of oxytocin resulted in a significant reduction in testicular and plasma testosterone levels throughout the 4-week period examined and, after 14 days of treatment, lipid droplets were seen in the Leydig cells of treated but not control animals. Concentrations of dihydrotestosterone in the plasma and testes of the oxytocin-treated animals, however, were significantly elevated after 7 and 14 days and at no time fell below control values. Plasma FSH levels were also lower in the oxytocin-treated animals. Intratesticular oxytocin treatment did not affect LH or oxytocin concentrations in the plasma, epididymal sperm counts or the number of Leydig cells in the testis. Empty Accurel devices had no effect on testicular morphology. This study provides the first evidence that oxytocin in vivo can modify steroidogenesis in the testis.


Axillary transvalvular microaxial pump as extended bridge to transcatheter aortic valve replacement in cardiogenic shock with severe aortic stenosis.
Cardiogenic shock in the setting of severe aortic stenosis is associated with poor outcomes. We describe 5 patients with cardiogenic shock and severe aortic stenosis who received an axillary microaxial pump (Impella) as an extended bridge to transcatheter aortic valve replacement. The median (range) age was 65 (61-87) years old, 80% were male, and 80% presented with stage D or E cardiogenic shock. In most cases, balloon aortic valvuloplasty was performed prior to pump insertion. Stabilization by Impella allowed for heart team evaluation and additional interventions, including percutaneous coronary intervention, MitraClip, and cardioversion. After a median (range) of 7 (5-14) days of Impella support, semi-elective transcatheter aortic valve replacement was successfully performed. All patients survived to discharge. Four patients (80%) were alive beyond 1 year. In these high-risk patients, prolonged support with a microaxial pump allowed for stabilization, ancillary interventions, and multi-disciplinary heart team evaluation prior to transcatheter aortic valve replacement.


Weather suitability for outdoor tourism in three European regions in first decades of the twenty-first century.
Outdoor tourism and recreational activities strongly depend on actual meteorological conditions. Traditionally, in three studied regions, the peak of tourists' streams concentrates in summer months. In the present study, we assess suitability of weather conditions for various forms of outdoor tourism in different regions of Serbia, Poland and Ukraine. Additionally, how the location of the station differentiates temporal patterns of weather suitability will be discussed. To analyse the suitability of weather conditions for various forms of outdoor recreation, we have chosen 23 meteorological stations of the national weather networks which represent different tourism areas and destinations. For each weather station, daily data for the period 2000-2017 of air temperature, relative humidity, total cloud cover and wind speed (at 10 m above ground) for 12 UTC as well as the daily maximum and minimum temperature, precipitation totals and snow cover depth were applied. Suitability of climate for outdoor recreation and tourism is assessed by the Weather Suitability Index (WSI) based on Błażejczyk's bio-thermal weather classification. The results of research show that passive forms of recreation (sun and air bathing) are preferred mostly in months from May till August or September. For the active forms of recreation, weather in summer months is very oppressive, especially in the resorts located in the south (Serbia, southern Ukraine). Active forms of recreation are preferred there in autumn, winter and spring months.


Submaximal cardiopulmonary exercise testing for the evaluation of unexplained dyspnea.
Gas exchange measurements obtained during submaximal exercise have been shown to provide prognostic and diagnostic information in patients with heart failure (HF) and to differentiate heart versus lung limitations in patients with unexplained dyspnea. The aim of our study was to assess the clinical utility of submaximal cardiopulmonary exercise testing using the Shape-HF equipment in identifying the cause of unexplained dyspnea. A total of 65 patients underwent Shape-HF tests from September 2010 to June 2011 for unexplained dyspnea at our center. Of 65 patients, 39 were men and 26 were women. In this study, 23 patients had preexisting asthma or chronic obstructive pulmonary disease (COPD); 19 patients had a pacemaker (8), an implantable cardioverter defibrillator (2), or a cardiac resynchronization therapy defibrillator (CRT-D) (9). The study revealed that submaximal cardiopulmonary exercise testing provided supportive clinical data for deconditioning, pulmonary limitations (eg, COPD, interstitial lung disease, sleep apnea), pulmonary hypertension, and chronotropic incompetence in 21.5%, 23.1%, 13.8%, and 6.2% of patients, respectively. Pulmonary hypertension was confirmed in 55% of patients by echocardiography and lung problems were confirmed in 40% of patients by pulmonary function test and sleep study. Of nine patients with an implanted CRT-D, optimization of atrioventricular and interventricular programming was performed in seven (78%) using gas exchange monitoring while performing a steady state, low-level treadmill walk. Submaximal cardiopulmonary exercise testing has strongly suggested the diagnosis of COPD, interstitial lung disease, pulmonary hypertension, and deconditioning and has led to appropriate testing. Based on prior studies, we also used Shape-HF for its approved purpose of optimizing CRT-D programming in patients with HF, leading to clinical improvement.


Pediatric aspects of inpatient health information technology systems.
In the past 3 years, the Health Information Technology for Economic and Clinical Health Act accelerated the adoption of electronic health records (EHRs) with providers and hospitals, who can claim incentive monies related to meaningful use. Despite the increase in adoption of commercial EHRs in pediatric settings, there has been little support for EHR tools and functionalities that promote pediatric quality improvement and patient safety, and children remain at higher risk than adults for medical errors in inpatient environments. Health information technology (HIT) tailored to the needs of pediatric health care providers can improve care by reducing the likelihood of errors through information assurance and minimizing the harm that results from errors. This technical report outlines pediatric-specific concepts, child health needs and their data elements, and required functionalities in inpatient clinical information systems that may be missing in adult-oriented HIT systems with negative consequences for pediatric inpatient care. It is imperative that inpatient (and outpatient) HIT systems be adapted to improve their ability to properly support safe health care delivery for children.


Antibody response of swine to outer membrane components of Haemophilus pleuropneumoniae during infection.
Sera from pigs infected with Haemophilus (Actinobacillus) pleuropneumoniae were tested for antibodies to outer membrane proteins (OMPs) of the organism by immunoblotting. Convalescent sera were produced in naturally born, colostrum-fed pigs and in cesarean-derived, colostrum-deprived pigs given H. pleuropneumoniae serotype 5 intranasally twice at 5-week intervals. Sera, collected at weekly intervals, were reacted with Sarkosyl-insoluble, OMP-enriched preparations of H. pleuropneumoniae which had been separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose. Antibodies were detected to OMPs with an apparent molecular weight of 16,500 (16.5K OMP); to 29K, 38.5K, 43.5K, 45K, 49.5K, and 66.5K OMPs; and to several high-molecular-weight (greater than or equal to 94,000) OMPs, but not to the major 42K OMP. Antibodies to the heat-modifiable OMP (29K/43.5K) and the 38.5K OMP were detected in sera from noninfected pigs. Antibodies were also detected to two broad 54,000- and 95,000-molecular-weight bands which did not stain with Coomassie blue, stained with silver nitrate, resisted proteinase K digestion, and were eliminated by oxidation with sodium metaperiodate. This indicates that the 54,000- and 95,000-molecular-weight bands represent polysaccharide, possibly capsular or lipopolysaccharide immunogens. Adsorption of sera with cells from the homologous serotype 5 strain removed antibodies to the 45K, 49.5K, 66.5K, and greater than or equal to 94K OMPs and to the two polysaccharide bands, indicating that these antibodies were directed primarily to surface-exposed epitopes. When tested with OMP preparations from other serotype 5 strains, heterogeneity was apparent, both in the reactions with OMPs and with the polysaccharide bands. Silver staining of proteinase K-treated, whole-cell lysates from serotype 5 strains also indicated variable expression of the polysaccharide bands. Sera also reacted with OMPs from H. pleuropneumoniae serotypes 1 and 7; however, several OMPs and the lipopolysaccharide or polysaccharide determinants of these serotypes appeared to be type specific.


RNA sequencing identifies common pathways between cigarette smoke exposure and replicative senescence in human airway epithelia.
Aging is affected by genetic and environmental factors, and cigarette smoking is strongly associated with accumulation of senescent cells. In this study, we wanted to identify genes that may potentially be beneficial for cell survival in response to cigarette smoke and thereby may contribute to development of cellular senescence. Primary human bronchial epithelial cells from five healthy donors were cultured, treated with or without 1.5% cigarette smoke extract (CSE) for 24 h or were passaged into replicative senescence. Transcriptome changes were monitored using RNA-seq in CSE and non-CSE exposed cells and those passaged into replicative senescence. We found that, among 1534 genes differentially regulated during senescence and 599 after CSE exposure, 243 were altered in both conditions, representing strong enrichment. Pathways and gene sets overrepresented in both conditions belonged to cellular processes that regulate reactive oxygen species, proteasome degradation, and NF-κB signaling. Our results offer insights into gene expression responses during cellular aging and cigarette smoke exposure, and identify potential molecular pathways that are altered by cigarette smoke and may also promote airway epithelial cell senescence.


[Local treatment for persistent inflammatory arthritis of the knee: intra-articular injection of glucocorticoids and bed rest].
Two studies evaluating treatment options for persistent mono-arthritis are discussed. In one study arthroscopic lavage plus intra-articular glucocorticoids was compared with arthroscopic lavage without glucocorticoids, and with joint aspiration plus glucocorticoids. Both intra-articular glucocorticoids and lavage have their merits, but glucocorticoids appear to be slightly more effective. In the second study radioactive Yttrium plus intra-articular glucocorticoids was compared with glucocorticoids alone. Yttrium provided little additional value. There are some indications that a period of strict bed rest after treatment is beneficial. Intra-articular glucocorticoids should not be administered more frequently than three times per year in the same joint.


[Genetic characterization of adenovirus isolated from follicular conjunctivitis and epidemic keratoconjunctivitis in a group of Mexican patients].
To determine the adenovirus serotype in Mexican patients with folicular conjunctivitis and keratoconjunctivitis. Adenovirus-specific PCR was used to analyze sample scrapings from the inferior fornix of patients with follicular conjunctivitis and clinical suspicion of adenovirus from January 2005 to December 2006. Identification of the serotype was made by automated sequencing. The nucleotide sequences obtained were compared with the reported sequences in GenBank. Descriptive statistical analyses were performed on the results. Of the 77 samples with clinical data of follicular conjunctivitis that were analyzed, 43 (56%) presented adenovirus. The sequencing of each positive sample allowed the identification of Ad1, Ad2, Ad3 and Ad8; the sequences of the serotype were identical those reported in GenBank with accession numbers: AF 534906 and AY 224420 for a sequence of the gene coding for the filament of Ad1 and Ad2 respectively, and AY 854180 and DQ 149614 for a sequence of the gene that codes for the Hexon protein of Ad3 and Ad8 respectively. From the statistical analysis it was possible to determine that a preferential seasonality of the serotype does not exist. In this work the Ad1, Ad2 and Ad3 serotypes were identified in patients with clinical diagnosis of follicular conjunctivitis in 2005. Ad2 was the predominant serotype. Ad8 was also detected in an outbreak of epidemic keratoconjunctivitis. From an epidemiological point of view, no serotype found seems to have a preferred seasonality.


SlARF4, an auxin response factor involved in the control of sugar metabolism during tomato fruit development.
Successful completion of fruit developmental programs depends on the interplay between multiple phytohormones. However, besides ethylene, the impact of other hormones on fruit quality traits remains elusive. A previous study has shown that down-regulation of SlARF4, a member of the tomato (Solanum lycopersicum) auxin response factor (ARF) gene family, results in a dark-green fruit phenotype with increased chloroplasts (Jones et al., 2002). This study further examines the role of this auxin transcriptional regulator during tomato fruit development at the level of transcripts, enzyme activities, and metabolites. It is noteworthy that the dark-green phenotype of antisense SlARF4-suppressed lines is restricted to fruit, suggesting that SlARF4 controls chlorophyll accumulation specifically in this organ. The SlARF4 underexpressing lines accumulate more starch at early stages of fruit development and display enhanced chlorophyll content and photochemical efficiency, which is consistent with the idea that fruit photosynthetic activity accounts for the elevated starch levels. SlARF4 expression is high in pericarp tissues of immature fruit and then undergoes a dramatic decline at the onset of ripening concomitant with the increase in sugar content. The higher starch content in developing fruits of SlARF4 down-regulated lines correlates with the up-regulation of genes and enzyme activities involved in starch biosynthesis, suggesting their negative regulation by SlARF4. Altogether, the data uncover the involvement of ARFs in the control of sugar content, an essential feature of fruit quality, and provide insight into the link between auxin signaling, chloroplastic activity, and sugar metabolism in developing fruit.


Football training improves lean body mass in men with prostate cancer undergoing androgen deprivation therapy.
Androgen deprivation therapy (ADT) remains a cornerstone in the management of patients with prostate cancer (PCa) despite adverse effects on body composition and functional parameters. We compared the effects of football training with standard care in PCa patients managed with ADT (> 6 months). Fifty-seven men aged 67 (range: 43-74) were randomly assigned to a football group (FG, n = 29) or a usual care control group (CON, n = 28). The primary outcome was change in lean body mass (LBM) assessed by dual-energy X-ray absorptiometry scanning. Secondary outcomes included changes in knee-extensor muscle strength (one repetition maximum), fat percentage, and maximal oxygen uptake (VO2max ). Mean heart rate during training was 137.7 (standard deviation 13.7) bpm or 84.6 (3.9)% HRmax. In FG, LBM increased by 0.5 kg [95% confidence interval (CI) 0.1-0.9; P = 0.02] with no change in CON (mean group difference 0.7 kg; 95% CI 0.1-1.2; P = 0.02). Also, muscle strength increased in FG (8.9 kg; 95% CI 6.0-11.8; P < 0.001) with no change in CON (mean group difference 6.7 kg; 95% CI 2.8-10.7; P < 0.001). In FG, VO2max increased (1.0 mL/kg/min; 95% CI 0.2-1.9; P = 0.02) and fat percentage tended to decrease (0.7%; 95%CI 1.3-0.0; P = 0.06), but these changes were not significantly different from CON. In conclusion, football training over 12 weeks improved LBM and muscle strength compared with usual care in men with prostate cancer receiving ADT.


Smoking patterns in students of higher institutions of learning in Nigeria.
A study of the pattern of cigarette smoking among 2317 students of both sexes attending some selected universities, polytechnics, colleges of education and schools of nursing in Nigeria was undertaken by means of questionnaires. It was observed that 436 (30%) of male students and 174 (21%) of female students smoked cigarettes. The prevalence was noted to be on the increase for female students when compared with previous studies carried out in similar institutions in Nigeria. The highest prevalence of smoking was among students from colleges of education. Over 50% of the students were mild smokers while less than 50% had exceeded a duration of 5 years of smoking. The latter finding suggests that it may still be feasible to convince a large proportion of them to stop smoking or to cut down on the number of cigarettes smoked.


A SPRY2 mutation leading to MAPK/ERK pathway inhibition is associated with an autosomal dominant form of IgA nephropathy.
IgA nephropathy (IgAN) represents the most common primary glomerulonephritis worldwide with a prevalence of 25-50% among patients with primary glomerulopathies. In ~5-10% of the patients the disease segregates with an autosomal dominant (AD) pattern. Association studies identified loci on chromosomes 1q32, 6p21, 8p23, 17p13, 22q12, whereas classical linkage studies on AD families identified loci on chromosomes 2q36, 4q26-31, 6q22, 17q12-22. We have studied a large Sicilian family where IgAN segregates with an AD transmission. To identify the causal gene, the exomes of two affected and one unaffected individual have been sequenced. From the bioinformatics analysis a p.(Arg119Trp) variant in the SPRY2 gene was identified as the probable disease-causing mutation. Moreover, functional characterization of this variant showed that it is responsible for the inhibition of the MAPK/ERK1/2 pathway. The same effect was observed in two sporadic IgAN patients carriers of wild-type SPRY2, suggesting that downregulation of the MAPK/ERK1/2 pathway represents a common mechanism leading to IgAN.


The interconversion of UDP-arabinopyranose and UDP-arabinofuranose is indispensable for plant development in Arabidopsis.
L-Ara, an important constituent of plant cell walls, is found predominantly in the furanose rather than in the thermodynamically more stable pyranose form. Nucleotide sugar mutases have been demonstrated to interconvert UDP-Larabinopyranose (UDP-Arap) and UDP-L-arabinofuranose (UDP-Araf) in rice (Oryza sativa). These enzymes belong to a small gene family encoding the previously named Reversibly Glycosylated Proteins (RGPs). RGPs are plant-specific cytosolic proteins that tend to associate with the endomembrane system. In Arabidopsis thaliana, the RGP protein family consists of five closely related members. We characterized all five RGPs regarding their expression pattern and subcellular localizations in transgenic Arabidopsis plants. Enzymatic activity assays of recombinant proteins expressed in Escherichia coli identified three of the Arabidopsis RGP protein family members as UDP-L-Ara mutases that catalyze the formation of UDP-Araf from UDP-Arap. Coimmunoprecipitation and subsequent liquid chromatography-electrospray ionization-tandem mass spectrometry analysis revealed a distinct interaction network between RGPs in different Arabidopsis organs. Examination of cell wall polysaccharide preparations from RGP1 and RGP2 knockout mutants showed a significant reduction in total L-Ara content (12–31%) compared with wild-type plants. Concomitant downregulation of RGP1 and RGP2 expression results in plants almost completely deficient in cell wall–derived L-Ara and exhibiting severe developmental defects.


Public awareness of warning signs and symptoms of cancer in oman: a community-based survey of adults.
The majority of deaths from cancer occur in low and middle income countries, partly due to poor public awareness of the signs and symptoms of cancer. A community based survey using the Cancer Awareness Measure (CAM) questionnaire was conducted in three different communities in Oman. Omani adults aged 18 years and above were invited to participate in the study. A total of 345 responded from 450 invited participants (response rate=76.7%). The majority of respondents were unable to identify the common signs and symptoms of cancer identified in the CAM (average awareness was 40.6%). The most emotional barrier to seeking help was worry about what the doctor might find (223, 64.6%); a practical barrier was too busy to make an appointment (259, 75.1%) and a service barrier was difficulty talking to the doctor (159, 46.1%). The majority of respondents (more than 60% for seven out of ten symptoms) would seek medical help in two weeks for most signs or symptoms of cancer. Females were significantly more likely than males to be embarrassed (p<0.001), scared (p=0.001), and lack confidence talking about their symptoms (p=0.022). Urgent strategies are needed to improve public awareness of the signs and symptoms of cancer in Oman. This might leads to earlier diagnosis, improved prognosis and reduced mortality from cancer.


Cold-Responsive Nanocapsules Enable the Sole-Cryoprotectant-Trehalose Cryopreservation of β Cell-Laden Hydrogels for Diabetes Treatment.
Islet transplantation has been one promising strategy in diabetes treatment, which can maintain patient's insulin level long-term and avoid periodical insulin injections. However, donor shortage and temporal mismatch between donors and recipients has limited its widespread use. Therefore, searching for islet substitutes and developing efficient cryopreservation technology (providing potential islet bank for transplantation on demand) is in great need. Herein, a novel cryopreservation method is developed for islet β cells by combining microfluidic encapsulation and cold-responsive nanocapsules (CR-NCs). The cryopreserved cell-laden hydrogels (calcium alginate hydrogel, CAH) can be transplanted for diabetes treatment. During the freezing process, trehalose is released inside β cells through the CR-NCs and serves as the sole cryoprotectant (CPA). Additionally, CAH helps cells to survive the freeze-thaw process and provide cells with a natural immune barrier in vivo. Different from traditional cryopreservation methods, this method combining the CR-NCs and hydrogel encapsulation replaces the toxic CPAs with natural trehalose. Great preservation results are obtained and transplantation experiments of diabetic rats further prove the excellent glucose regulation ability of such β cell-laden hydrogels post cryopreservation. This novel cryopreservation method helps to establish a reliable and ready-to-use bank of biological samples for transplantation therapy and other biomedical applications.


Acholeplasma laidlawii cells acutely and chronically infected with group 1 acholeplasmavirus.
Interactions between group 1 acholeplasmaviruses and their host cells were studied. Acutely infected, chronically infected and uninfected cultures of Acholeplasma laidlawii strain JA1 were compared by their growth in broth and on agar, by the sensitivities of the uninfected and chronically infected cells to representatives of each of the three groups of acholeplasmaviruses, and by their SDS-PAGE polypeptide profiles. Acutely infected cells resembled uninfected cells by these criteria, except for the fact that progeny virus was being released. Two types of chronically infected cells were found:rapid growers (the same doubling time as uninfected cells) and slow growers. The latter resembled uninfected cells, except for their slower growth and low-level release of virus, and the former was resistant to group 1 viruses and had a unique polypeptide profile. These biological characterizations help to establish the non-lytic, non-cytocidal cycle of the group 1 acholeplasmaviruses.


Npc1 gene mutation abnormally activates the classical Wnt signalling pathway in mouse kidneys and promotes renal fibrosis.
Niemann-Pick disease type C1 (NPC1) is a lysosomal lipid storage disease caused by NPC1 gene mutation. Our previous study found that, compared with wild-type (Npc1+/+ ) mice, the renal volume and weight of Npc1 gene mutant (Npc1-/- ) mice were significantly reduced. We speculate that Npc1 gene mutations may affect the basic structure of the kidneys of Npc1-/- mice, and thus affect their function. Therefore, we randomly selected postnatal Day 28 (P28) and P56 Npc1+/+ and Npc1-/- mice, and observed the renal structure and pathological changes by haematoxylin-eosin staining. The level of renal fibrosis was detected by immunofluorescence histochemical techniques, and western blotting was used to detect the expression levels of apoptosis-related proteins and canonical Wnt signalling pathway related proteins. The results showed that compared with Npc1+/+ mice, the kidneys of P28 and P56 Npc1-/- mice underwent apoptosis and fibrosis; furthermore, there were obvious vacuoles in the cytoplasm of renal tubular epithelial cells of P56 Npc1-/- mice, the cell bodies were loose and foam-like, and the canonical Wnt signalling pathway was abnormally activated. These results showed that Npc1 gene mutation can cause pathological changes in the kidneys of mice. As age increased, vacuoles developed in the cytoplasm of renal tubular epithelial cells, and apoptosis of renal cells, abnormal activation of the Wnt signalling pathway, and promotion of renal fibrosis increased.


Generic first-order phase transitions between isotropic and orientational phases with polyhedral symmetries.
Polyhedral nematics are examples of exotic orientational phases that possess a complex internal symmetry, representing highly nontrivial ways of rotational symmetry breaking, and are subject to current experimental pursuits in colloidal and molecular systems. The classification of these phases has been known for a long time; however, their transitions to the disordered isotropic liquid phase remain largely unexplored, except for a few symmetries. In this work, we utilize a recently introduced non-Abelian gauge theory to explore the nature of the underlying nematic-isotropic transition for all three-dimensional polyhedral nematics. The gauge theory can readily be applied to nematic phases with an arbitrary point-group symmetry, including those where traditional Landau methods and the associated lattice models may become too involved to implement owing to a prohibitive order-parameter tensor of high rank or (the absence of) mirror symmetries. By means of exhaustive Monte Carlo simulations, we find that the nematic-isotropic transition is generically first-order for all polyhedral symmetries. Moreover, we show that this universal result is fully consistent with our expectation from a renormalization group approach, as well as with other lattice models for symmetries already studied in the literature. We argue that extreme fine tuning is required to promote those transitions to second-order ones. We also comment on the nature of phase transitions breaking the O(3) symmetry in general cases.


Lipoprotein lipase and atherosclerosis.
Lipoprotein lipase (LPL) is a rate-limiting enzyme that hydrolyzes circulating triglyceride-rich lipoprotein such as very low density lipoproteins and chylomicrons. A decrease in LPL activity is associated with an increase in plasma triglycerides (TG) and decrease in high density lipoprotein (HDL) cholesterol. The increase in plasma TG and decrease in HDL cholesterol are risk factors of coronary heart disease. However, whether LPL directly or indirectly promotes or protects against atherosclerosis remains unclear as two contrary views exist in this regard: one where LPL promotes atherosclerosis and one where LPL protects against atherosclerosis. Many studies have been carried out to investigate whether LPL is an anti-atherogenic or atherogenic enzyme by using animals with genetic defects or with an excess of this enzyme. From these studies, much evidence has been acquired showing that LPL is an anti-atherogenic enzyme. We hypothesized that elevating LPL activity would cause a reduction of plasma TG and increase in HDL cholesterol, resulting in protection against the development of atherosclerosis. To test this hypothesis, we studied the effects of the LPL activator NO-1886 in animals. NO-1886 has been shown to increase LPL mRNA in adipose tissue and myocardium, and increase LPL activity in adipose tissue, myocardium and skeletal muscle, resulting in an elevation of postheparin plasma LPL activity and LPL mass in rats. NO-1886 has also been shown to decrease plasma TG levels accompanied by a concomitant rise in HDL cholesterol. Long-term administration of NO-1886 to rats and rabbits with experimental atherosclerosis inhibited the development of atherosclerotic lesions in coronary arteries and aortae. The results of multiple regression analysis in these studies suggest that the increase in plasma HDL cholesterol and the decrease in TG protect against atherosclerosis. We have determined in our studies that the atherogenic lipid profile is changed to an anti-atherogenic lipid profile by increasing LPL activity, resulting in protection against the development of atherosclerosis. Therefore, we believe that high activity of LPL is anti-atherogenic, whereas a low activity of LPL is atherogenic.


American Clinical Neurophysiology Society Guideline 7: Guidelines for EEG Reporting.
This EEG Guideline incorporates the practice of structuring a report of results obtained during routine adult electroencephalography. It is intended to reflect one of the current practices in reporting an EEG and serves as a revision of the previous guideline entitled "Writing an EEG Report." The goal of this guideline is not only to convey clinically relevant information, but also to improve interrater reliability for clinical and research use by standardizing the format of EEG reports. With this in mind, there is expanded documentation of the patient history to include more relevant clinical information that can affect the EEG recording and interpretation. Recommendations for the technical conditions of the recording are also enhanced to include post hoc review parameters and type of EEG recording. Sleep feature documentation is also expanded upon. More descriptive terms are included for background features and interictal discharges that are concordant with efforts to standardize terminology. In the clinical correlation section, examples of common clinical scenarios are now provided that encourages uniformity in reporting. Including digital samples of abnormal waveforms is now readily available with current EEG recording systems and may be beneficial in augmenting reports when controversial waveforms or important features are encountered.


Copy-back viral genomes induce a cellular stress response that interferes with viral protein expression without affecting antiviral immunity.
Antiviral responses are often accompanied by translation inhibition and formation of stress granules (SG) in infected cells. However, the triggers for these processes and their role during infection remain subjects of active investigation. Copy-back viral genomes (cbVGs) are the primary inducers of the Mitochondrial Antiviral Signaling (MAVS) pathway and antiviral immunity during Sendai Virus (SeV) and Respiratory Syncytial virus (RSV) infections. The relationship between cbVGs and cellular stress during viral infections is unknown. Here we show that SG form during infections containing high levels of cbVGs, and not during infections with low levels of cbVGs. Moreover, using RNA fluorescent in situ hybridization to differentiate accumulation of standard viral genomes from cbVGs at a single-cell level during infection, we show that SG form exclusively in cells that accumulate high levels of cbVGs. PKR activation is increased during high cbVG infections and, as expected, PKR is necessary to induce virus-induced SG. However, SG form independent of MAVS signaling, demonstrating that cbVGs induce antiviral immunity and SG formation through two independent mechanisms. Furthermore, we show that translation inhibition and SG formation do not affect the overall expression of interferon and interferon stimulated genes during infection, making the stress response dispensable for antiviral immunity. Using live-cell imaging, we show that SG formation is highly dynamic and correlates with a drastic reduction of viral protein expression even in cells infected for several days. Through analysis of active protein translation at a single cell level, we show that infected cells that form SG show inhibition of protein translation. Together, our data reveal a new cbVG-driven mechanism of viral interference where cbVGs induce PKR-mediated translation inhibition and SG formation leading to a reduction in viral protein expression without altering overall antiviral immunity.


Voltammetric demonstration of thermally induced natural convection in aqueous solution.
In electrochemical systems imperfect thermostating inevitably leads to the presence of bulk convective flows. As recognised by Nernst [Z. Phys. Chem., 1904, 52] damping of these bulk convective flows next to a solid surface, or at the electrode, leads to diffusional mass transport predominating locally. This work questions the exclusivity of diffusional transport and provides hitherto unexplored physical insights into how thermally induced flows in bulk solution can, on both macro- and microelectrodes, influence a voltammetric measurement. Imperfect thermostating results in flows in the bulk solution which are predicted and here expeimentally shown to be of the order of 100 μm s-1. Here we show that even in the absence of natural convective flows induced by the electrochemical reaction itself, this thermally induced bulk convection can significantly affect the voltammetric response. First, evaporative losses from an open electrochemical cell can be sufficient to produce convective flows that can alter the electrochemical response. Second, electrodes with various sizes and geometries have been investigated and experimental results evidence that the sensitivity of an electrode to these flows in bulk solution is to a large extent controlled by the size of the surrounding non-conductive supporting substrate used to insulate parts of the electrode.


Rapid, simple serodiagnosis of murine typhus.
The dot-blot enzyme-linked immunosorbent assay (dot-ELISA) was compared to latex agglutination (LA) and the Weil-Felix OX-19 test for the diagnosis of murine typhus using the indirect immunofluorescent antibody (IFA) test as the 'gold standard'. With a panel of 74 positive and 47 negative sera, the dot-ELISA was 98% specific and 74% sensitive at a cut-off value of the second dot. With acute sera, latex agglutination was 100% specific and 74% sensitive at a cut-off titre of > or = 1:64. Both tests were more sensitive than the OX-19 test, which was 98% specific and 56% sensitive at a titre of > or = 1:320. Both dot-ELISA and latex agglutination were comparable and the results were available within one hour of testing. The rapidity, ease in performance and minimal requirement for electrical instruments made these 2 tests suitable for the diagnosis of murine typhus in countries where sophisticated laboratory facilities are lacking.


[Diagnostic value of determination of the blood concentrations of serotonin and high-sensitivity C-reactive protein in patients with tick-borne encephalitis].
The paper shows the informative value of changes in the serum levels of serotonin and high-sensitivity C-reactive protein in 44 patients in the acute period of tickborne encephalitis. Group 1 included 35 patients with non-paralytic neuroinfection; Group 2 comprised 9 patients with paralytic neuroinfection. The serum level of high-sensitivity C-reactive protein increased and that of serotonin decreased in relation to the form and severity of the disease at its height. The elevated level of the former and the low concentration of the latter may serve as an early predictor of the development of a severe paralytic form. Investigating their serum concentrations should be included into an examination plan for patients with acute tick-borne encephalitis.


Subsequent maternal separation exacerbates neurobehavioral abnormalities in rats neonatally exposed to sevoflurane anesthesia.
Several recent studies suggest that in the human population, a routine, short anesthetic in otherwise healthy infants is void of neurodevelopmental insult. On the other hand, many human retrospective epidemiological studies report evidence of cognitive abnormalities in children after testing those who had different anesthesia-requiring procedures in early childhood. We tested in a rat model whether post-anesthesia stressful environmental factors can contribute to developmental abnormalities that were initiated by a relatively short exposure to sevoflurane, the most widely used anesthetic in pediatric anesthesia, whose polyvalent actions include enhancement of gamma-aminobutyric acid type A receptor (GABAAR) activity. Postnatal day 6 (P6) male Sprague-Dawley rats were anesthetized with sevoflurane for 60min. To simulate subsequent stress, the animals were subjected to a single maternal separation for 180min at P10. To study the role of GABAAR-mediated depolarization, subgroups of P6 rats received a single injection of the Na+-K+-2Cl- (NKCC1) inhibitor, bumetanide, prior to initiation of anesthesia with sevoflurane. Rats that were exposed to sevoflurane had decreased hypothalamic K+-2Cl- (KCC2) mRNA level (F(2,13)=3.839, P=0.049), increased NKCC1/KCC2 mRNA ratio (F(2,13)=5.043, P=0.024) and increased corticotropin-releasing hormone (CRH) mRNA level (F(2,12)=9.450, P=0.003) at P10, the age at which maternal separation was imposed. Adult rats, neonatally exposed to a combination of sevoflurane and maternal separation, exhibited increases in the escape latencies greater than animals exposed to sevoflurane only (P=0.012), and only rats in the sevoflurane plus maternal separation group spent significantly less time in the target quadrant during the Morris water maze test (F(4,55)=4.856, P=0.002). Bumetanide ameliorated abnormalities induced by sevoflurane and a combination of sevoflurane plus maternal separation. Neonatal exposure to sevoflurane may sensitize to stressors later in life, and post-exposure stress may exacerbate neurodevelopmental abnormalities even after a relatively short exposure to sevoflurane in rodents. The NKCC1 downregulation prior to exposure to the anesthetic may be therapeutic.


Recent advances in medicinal chemistry and pharmaceutical technology--strategies for drug delivery to the brain.
This paper provides a mini-review of some recent approaches for the treatment of brain pathologies examining both medicinal chemistry and pharmaceutical technology contributions. Medicinal chemistry-based strategies are essentially aimed at the chemical modification of low molecular weight drugs in order to increase their lipophilicity or the design of appropriate prodrugs, although this review will focus primarily on the use of prodrugs and not analog development. Recently, interest has been focused on the design and evaluation of prodrugs that are capable of exploiting one or more of the various endogenous transport systems at the level of the blood brain barrier (BBB). The technological strategies are essentially non-invasive methods of drug delivery to malignancies of the central nervous system (CNS) and are based on the use of nanosystems (colloidal carriers) such as liposomes, polymeric nanoparticles, solid lipid nanoparticles, polymeric micelles and dendrimers. The biodistribution of these nanocarriers can be manipulated by modifying their surface physico-chemical properties or by coating them with surfactants and polyethylene-glycols (PEGs). Liposomes, surfactant coated polymeric nanoparticles, and solid lipid nanoparticles are promising systems for delivery of drugs to tumors of the CNS. This mini-review discusses issues concerning the scope and limitations of both the medicinal chemistry and technological approaches. Based on the current findings, it can be concluded that crossing of the BBB and drug delivery to CNS is extremely complex and requires a multidisciplinary approach such as a close collaboration and common efforts among researchers of several scientific areas, particularly medicinal chemists, biologists and pharmaceutical technologists.


Intranasal immunization with polymer-grafted microparticles activates the nasal-associated lymphoid tissue and draining lymph nodes.
Waldeyer's ring is located at the juncture of the respiratory and alimentary tracts, where it is bombarded by inhaled and ingested antigens. However, knowledge of its exact function or consequences of its removal is incomplete. Recently, the murine nasal-associated lymphoid tissue (NALT) has been reported to have functional similarities to Waldeyer's ring and, thus, might be a suitable model to examine the function of oronasopharyngeal lymphoid tissues. To explore the capability of NALT to incite local mucosal and systemic immunity, we immunized mice intranasally (i.n.) with 3-(triethoxysilyl)-propyl-terminated polydimethylsiloxane (TS-PDMS)-grafted microparticles (MP), an inoculant previously shown to induce robust systemic and mucosal humoral immunity following intragastric (i.g.) administration. We demonstrated that i.n. immunization with low doses of microentrapped, but not soluble, human serum albumin (HSA) evoked robust circulating IgG responses (P < 0.05). Additionally, NALT cells isolated from MP-treated mice proliferated in vitro when restimulated with HSA (P < 0.05), suggesting that i.n. immunization with HSA-containing MP incited specific immunity via NALT cell activation. Coinciding with these observations, after i.n. MP administration HSA-specific spot-forming cells (SFC) were observed in NALT, and later posterior cervical lymph nodes (pCLN) and spleen (SPL), suggesting that the observed MP-induced specific systemic antibody responses emanated from the NALT. We also showed that i.n. immunization with HSA-containing TS-PDMS-grafted MP stimulated interleukin-4 (IL-4)-secreting lymphocytes in the NALT. This cytokine microenvironment was probably responsible for driving the IgG1 sera response observed after i.n. MP administration, via the migration of NALT-derived IgG1-committed B cells. Interestingly, unlike i.g. MP administration, i.n. immunization with HSA-containing MP did not evoke detectable specific IgA in any lymphoid tissue examined, or in nasal secretions, probably reflecting differences between NALT and other mucosae-associated lymphoid tissues (MALT).


Peripheral oxytocin treatment ameliorates obesity by reducing food intake and visceral fat mass.
Recent studies suggest that oxytocin (Oxt) is implicated in energy metabolism. We aimed to explore acute and sub-chronic effects of peripheral Oxt treatment via different routes on food intake and energy balance. Intraperitoneal (ip) injection of Oxt concentration-dependently decreased food intake in mice. Ip Oxt injection induced c-Fos expression in the hypothalamus and brain stem including arcuate nucleus (ARC), paraventricular nucleus (PVN) and nucleus tractus solitarius (NTS). Subcutaneous (sc) injection of Oxt suppressed food intake in normal and high fat diet-induced obese (DIO) mice. Daily sc injection of Oxt for 17 days in DIO mice reduced food intake for 6 days and body weight for the entire treatment period and additional 9 days after terminating Oxt. Oxt infusion by sc implanted osmotic minipumps for 13 days in DIO mice reduced food intake, body weight, and visceral fat mass and adipocyte size. Oxt infusion also decreased respiratory quotient specifically in light phase, ameliorated fatty liver and glucose intolerance, without affecting normal blood pressure in DIO mice. These results demonstrate that peripheral Oxt treatment reduces food intake and visceral fat mass, and ameliorates obesity, fatty liver and glucose intolerance. Peripheral Oxt treatment provides a new therapeutic avenue for treating obesity and hyperphagia.


Human lipocalin-1 association with 3H-testosterone and 3H-estradiol.
Topical androgens and estrogens have been studied for use in treating ocular conditions such as dry eye. The aim of this study was to identify proteins from normal human tears that associated with exogenously added sex steroid hormones. One of the major proteins in ocular tears is lipocalin-1. It binds a variety of lipids and other hydrophobic molecules and is proposed to function as a carrier protein or a lipid scavenger. Normal human tears were incubated with (3)H-testosterone or (3)H-estradiol. Labeled tear proteins were separated on a Q Sepharose Fast Flow (QFF) Hi Trap strong anion exchange column with a step gradient of NaCl. (3)H-testosterone or (3)H-estradiol was measured in aliquots of eluted fractions using scintillation counts, and the remainder of each sample was gel electrophoresed and silver stained. In separate experiments, (3)H-steroid-labeled tear proteins were electrophoresed in 15% polyacrylamide gels and excised from the gels. Tritium content of the proteins was measured in a scintillation counter. Immunoblots with antibodies to lipocalin-1 verified the migration of lipocalin-1 in the gels. (3)H-steroid labeled tear proteins were found in the 0.15 M NaCl fractions of QFF strong anion exchange columns. 18 kD lipocalin-1 (among other tear proteins) eluted in the 0.15 M NaCl fraction. Excision of labeled tear proteins from 15% polyacrylamide gels indicated that radioactive label was associated with an 18 kD protein. Immunoblots verified that lipocalin-1 migrated as an 18 kD protein. The sex steroid hormones testosterone and estradiol associated with 18 kD lipocalin-1 in human tears.


Purple sweet potato color protects against hepatocyte apoptosis through Sirt1 activation in high-fat-diet-treated mice.
Recent evidence indicates that the inhibition of hepatocyte apoptosis is possible to develop a potential therapeutic strategy for nonalcoholic fatty liver disease (NAFLD). Our previous work suggested that purple sweet potato color (PSPC), a class of naturally occurring anthocyanins, effectively improved many features of high-fat diet (HFD)-induced NAFLD. However, whether PSPC ameliorates HFD-induced hepatocyte apoptosis has never been investigated. Here we investigated the effects of PSPC on HFD-induced hepatic apoptosis and the mechanisms underlying these effects. Mice were divided into four groups: Control group, HFD group, HFD + PSPC group and PSPC group. PSPC was administered by daily oral gavage at doses of 700 mg/kg/day for 20 weeks. EX-527 (a SirT1-selective inhibitor) and Sirt1 siRNA were used to demonstrate the Sirt1 dependence of PSPC-mediated effects on apoptotic and survival signaling pathways in vivo and in vitro. Our results showed that PSPC reduced body weights, hepatic triglyceride contents, histopathological lesions and serum ALT levels in a mouse model of NAFLD induced by HFD. Furthermore, PSPC attenuated HFD-induced hepatocyte apoptosis ratio from 7.27 ± 0.92% to 1.79 ± 0.27% in mouse livers, which is insignificant compared with that of controls. Moreover, PSPC activated Sirt1 by boosting NAD+ level in HFD-treated mouse livers. Furthermore, PSPC promoted Sirt1-dependent suppression of P53-mediated apoptotic signaling and activation of Akt survival signaling pathway in HFD-treated mouse livers, which was confirmed by EX527 treatment. Moreover, Sirt1 knockdown abolished these ameliorative effects of PSPC on apoptosis and P53 acetylation and protein expression in PA-treated L02 cells. Ultimately, PSPC reduced Caspase-3 activation and Bax level, and elevated the Bcl-2 level in HFD-treated mouse livers. PSPC protected against HFD-induced hepatic apoptosis by promoting Sirt1- dependent inhibition of p53-apoptotic pathway and facilitation of Akt survival pathway. This study indicates that PSPC is a candidate for nutritional intervention of NAFLD.