Inhibition of reactive oxygen species production ameliorates inflammation induced by influenza A viruses via upregulation of SOCS1 and SOCS3.
Highly pathogenic avian influenza virus infection is associated with severe mortality in both humans and poultry. The mechanisms of disease pathogenesis and immunity are poorly understood although recent evidence suggests that cytokine/chemokine dysregulation contributes to disease severity following H5N1 infection. Influenza A virus infection causes a rapid influx of inflammatory cells, resulting in increased reactive oxygen species production, cytokine expression, and acute lung injury. Proinflammatory stimuli are known to induce intracellular reactive oxygen species by activating NADPH oxidase activity. We therefore hypothesized that inhibition of this activity would restore host cytokine homeostasis following avian influenza virus infection. A panel of airway epithelial and immune cells from mammalian and avian species were infected with A/Puerto Rico/8/1934 H1N1 virus, low-pathogenicity avian influenza H5N3 virus (A/duck/Victoria/0305-2/2012), highly pathogenic avian influenza H5N1 virus (A/chicken/Vietnam/0008/2004), or low-pathogenicity avian influenza H7N9 virus (A/Anhui/1/2013). Quantitative real-time reverse transcriptase PCR showed that H5N1 and H7N9 viruses significantly stimulated cytokine (interleukin-6, beta interferon, CXCL10, and CCL5) production. Among the influenza-induced cytokines, CCL5 was identified as a potential marker for overactive immunity. Apocynin, a Nox2 inhibitor, inhibited influenza-induced cytokines and reactive oxygen species production, although viral replication was not significantly altered in vitro. Interestingly, apocynin treatment significantly increased influenza virus-induced mRNA and protein expression of SOCS1 and SOCS3, enhancing negative regulation of cytokine signaling. These findings suggest that apocynin or its derivatives (targeting host responses) could be used in combination with antiviral strategies (targeting viruses) as therapeutic agents to ameliorate disease severity in susceptible species. Highly pathogenic avian influenza virus infection causes severe morbidity and mortality in both humans and poultry. Wide-spread antiviral resistance necessitates the need for the development of additional novel therapeutic measures to modulate overactive host immune responses after infection. Disease severity following avian influenza virus infection can be attributed in part to hyperinduction of inflammatory mediators such as cytokines, chemokines, and reactive oxygen species. Our study shows that highly pathogenic avian influenza H5N1 virus and low-pathogenicity avian influenza H7N9 virus (both associated with human fatalities) promote inactivation of FoxO3 and downregulation of the TAM receptor tyrosine kinase, Tyro3, leading to augmentation of the inflammatory cytokine response. Inhibition of influenza-induced reactive oxygen species with apocynin activated FoxO3 and stimulated SOCS1 and SOCS3 proteins, restoring cytokine homeostasis. We conclude that modulation of host immune responses with antioxidant and/or anti-inflammatory agents in combination with antiviral therapy may have important therapeutic benefits.


Acute Respiratory Failure Post Single Lung Transplantation: An Unusual Cause.
Vascular complications of pulmonary vessels post-transplant are rare but are associated with high morbidity and mortality. We describe the first published case of surgical management of pulmonary vein stenosis complicated by early thrombosis post single lung transplantation. This report describes early diagnostic techniques and subsequent management options in such patients where pulmonary vascular complications are a potential.


Group counseling/psychotherapy as a school intervention to enhance close friendships in preadolescence.
This article recognizes the development of a close same-sex friendship as a developmental task, distinguishable from general successful peer relations. It is argued that a close friendship includes the component of relational competence, which is effectively developed in interactional group psychotherapy and therefore may be a viable method to promote intimate friendships in preadolescence. The rationale for linking growth in intimate friendship with group psychotherapy is provided and supported with illustrations of the therapeutic process as well as with research data.


Cobalt(iii)-catalyzed redox-neutral [4+2]-annulation of N-chlorobenzamides/acrylamides with alkylidenecyclopropanes at room temperature.
An efficient synthesis of substituted 3,4-dihydroisoquinolinones through [4+2]-annulation of N-chlorobenzamides/acrylamides having a monodentate directing group with alkylidenecyclopropanes in the presence of a less expensive, highly abundant and air stable Co(iii) catalyst via a C-H activation is demonstrated. In this reaction, the N-Cl bond of N-chlorobenzamide serves as an internal oxidant and thus an external metal oxidant is avoided. The 3,4-dihydroisoquinolinone derivatives are converted successfully into the highly useful imidoyl chloride derivatives. The deuterium labeling and kinetic isolabelling studies reveal that the C-H activation is a rate-determining step in this cyclization reaction.


Fluoride toxicity and status of serum thyroid hormones, brain histopathology, and learning memory in rats: a multigenerational assessment.
High-fluoride (100 and 200 ppm) water was administered to rats orally to study the fluoride-induced changes on the thyroid hormone status, the histopathology of discrete brain regions, the acetylcholine esterase activity, and the learning and memory abilities in multigeneration rats. Significant decrease in the serum-free thyroxine (FT4) and free triiodothyronine (FT3) levels and decrease in acetylcholine esterase activity in fluoride-treated group were observed. Presence of eosinophilic Purkinje cells, degenerating neurons, decreased granular cells, and vacuolations were noted in discrete brain regions of the fluoride-treated group. In the T-maze experiments, the fluoride-treated group showed poor acquisition and retention and higher latency when compared with the control. The alterations were more profound in the third generation when compared with the first- and second-generation fluoride-treated group. Changes in the thyroid hormone levels in the present study might have imbalanced the oxidant/antioxidant system, which further led to a reduction in learning memory ability. Hence, presence of generational or cumulative effects of fluoride on the development of the offspring when it is ingested continuously through multiple generations is evident from the present study.


Mapping of heparin/heparan sulfate binding sites on αvβ3 integrin by molecular docking.
Heparin/heparan sulfate interact with growth factors, chemokines, extracellular proteins, and receptors. Integrins are αβ heterodimers that serve as receptors for extracellular proteins, regulate cell behavior, and participate in extracellular matrix assembly. Heparin binds to RGD-dependent integrins (αIIbβ3, α5β1, αvβ3, and αvβ5) and to RGD-independent integrins (α4β1, αXβ2, and αMβ2), but their binding sites have not been located on integrins. We report the mapping of heparin binding sites on the ectodomain of αvβ3 integrin by molecular modeling. The surface of the ectodomain was scanned with small rigid probes mimicking the sulfated domains of heparan sulfate. Docking results were clustered into binding spots. The best results were selected for further docking simulations with heparin hexasaccharide. Six potential binding spots containing lysine and/or arginine residues were identified on the ectodomain of αvβ3 integrin. Heparin would mostly bind to the top of the genu domain, the Calf-I domain of the α subunit, and the top of the β subunit of RGD-dependent integrins. Three spots were close enough from each other on the integrin surface to form an extended binding site that could interact with heparin/heparan sulfate chains. Because heparin does not bind to the same integrin site as protein ligands, no steric hindrance prevents the formation of ternary complexes comprising the integrin, its protein ligand, and heparin/heparan sulfate. The basic amino acid residues predicted to interact with heparin are conserved in the sequences of RGD-dependent but not of RGD-independent integrins suggesting that heparin/heparan sulfate could bind to different sites on these two integrin subfamilies.


Measurements of nonlinearity parameter B/A of water-saturated glass beads under uniaxial compressive loading.
This Letter reports measurements of nonlinearity parameter B/A of water-saturated glass beads under uniaxial compression representative of the gravitational loading in seabed. The finite-amplitude method in conjunction with the multi-Gaussian beam model is used to determine B/A, in which the use of an accurate source function is found to be important in achieving the overall measurement accuracy. The estimated B/A is on the order of a hundred and decreases with increasing uniaxial compression up to 50 kPa, so that B/A could change as much as 20% over a depth of 5 m for a typical coarse sandy sediment.


Atypical methylmalonic aciduria: frequency of mutations in the methylmalonyl CoA epimerase gene (MCEE).
Methylmalonic aciduria is known to result from defects in the enzyme methylmalonyl CoA mutase (MCM) (mut complementation group) and from defects in the synthesis of the MCM cofactor adenosylcobalamin (cblA, cblB, cblC, cblD, and cblF groups). Two patients who excrete methylmalonic acid have recently been shown to have a homozygous nonsense mutation in the gene coding for methylmalonyl CoA epimerase (MCEE). To further understand the cause of methylmalonic acid excretion, the MCEE gene was sequenced in 229 patients with elevations of methylmalonic acid excretion for which no cause was known. Mutations in MCEE were detected in five patients: two patients homozygous for c.139C>T, p.R47X, one patient homozygous for c.178A>C, p.K60Q, and two patients heterozygous for c.427C>T, p.R143C. Fusion of fibroblast lines from two patients homozygous for c.139C>T, p.R47X did not result in correction of [(14)C]propionate incorporation toward control values while the defect in these fibroblasts was complemented by mut, cblA, and cblB fibroblasts. Infection with wild-type MCEE cDNA resulted in correction of the biochemical phenotype in cells from both patients.


The inhibitory effect of terfenadine and flurbiprofen on early and late-phase bronchoconstriction following allergen challenge in atopic asthma.
We have studied the effect of cyclo-oxygenase inhibition and H1-receptor antagonism on the early and late bronchoconstrictor responses to inhaled allergen in mild atopic asthmatics. In the first phase of the study histamine inhalation challenge tests were performed in seven mild, atopic asthmatics 2 h after treatment with placebo or flurbiprofen (50, 100 or 150 mg). Flurbiprofen in these single doses had no effect on histamine reactivity. Ten atopic asthmatics participated in the second phase of the study in which the time course of the bronchoconstrictor response to inhalation of allergen was observed on four separate occasions after treatment with (a) placebo, (b) flurbiprofen, 150 mg, (c) terfenadine 180 mg, and (d) the combination of flurbiprofen and terfenadine. On each occasion subjects inhaled a concentration of allergen (Dermatagaphoides pteronyssinus, grass pollen) that had previously been shown to produce a 30% fall in FEV1 (PC30 allergen). The mean maximum fall in FEV1 during the early reaction was 33.2 +/- 3.3% from the post-saline baseline value following placebo and this was reduced to 27.5 +/- 5.3% after flurbiprofen (n.s.), 20.3 +/- 3.2% after terfenadine (P less than 0.05), and 23.1 +/- 2.3 after the treatment combination (P less than 0.05). Seven subjects developed late asthmatic reactions (LAR) after placebo and in these subjects the mean maximum fall in PEFR during the LAR was reduced from 22.6 +/- 3.1% after placebo to 16.7 +/- 3.2% after flurbiprofen (P less than 0.05), 15.2 +/- 2.3% after terfenadine (P less than 0.05) and 11.5 +/- 3.1% after the treatment combination (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Phytochemical-Based Nanocomposites for the Treatment of Bacterial Biofilms.
Biofilm infections are responsible for at least 65% of human bacterial infections. These biofilms are refractory to conventional antibiotics, leading to chronic infections and nonhealing wounds. Plant-derived antibiotics (phytochemicals) are promising alternative antimicrobial treatments featuring antimicrobial properties. However, their poor solubility in aqueous media limits their application in treating biofilm infections. Phytochemicals were incorporated into cross-linked polymer nanocomposite "sponges" for the treatment of bacterial biofilms. The results indicated encapsulating low log P phytochemicals effectively eliminated biofilms while demonstrating low cytotoxicity against mammalian fibroblast cells.


Detecting differential patterns of interaction in molecular pathways.
We consider statistical inference for potentially heterogeneous patterns of association characterizing the expression of bio-molecular pathways across different biologic conditions. We discuss a modeling approach based on Gaussian-directed acyclic graphs and provide computational and methodological details needed for posterior inference. Our application finds motivation in reverse phase protein array data from a study on acute myeloid leukemia, where interest centers on contrasting refractory versus relapsed patients. We illustrate the proposed method through both synthetic and case study data.


An immunohistochemical study of the effects of orexin receptor blockade on phospholipase C-β3 level in rat hippocampal dentate gyrus neurons.
Orexin-A (hypocretin-1) is a neuropeptide that is produced in the lateral hypothalamic area (LHA) and promotes widespread cortical activation. We investigated the effect of SB-334867, a selective orexin receptor 1 (OXR1) antagonist, on phospholipase C-β3 (PLCβ3) level using slices of rat hippocampus preparations and immunohistochemistry. We used three Wistar rats in each of three groups. The control group was untreated rats and SB vehicle and SB groups received SB vehicle and 10 mg/kg SB-334867 daily from postnatal day (PND) 12 to PND30, respectively. We found that the orexin receptor antagonist decreased the PLCβ3 level in the inner and outer blades of dentate gyrus (DG) compared to SB vehicle treated rats. Orexin may increase the PLCβ3 level in most regions of the rat hippocampus.


Spectrum-Effect Relationship Analysis of Bioactive Compounds in Zanthoxylum nitidum (Roxb.) DC. by Ultra-High Performance Liquid Chromatography Mass Spectrometry Coupled With Comprehensive Filtering Approaches.
Zanthoxylum nitidum (Roxb.) DC. (ZN), with strong effects of anti-inflammation and antioxidant activities is treated as a core herb in traditional Chinese medicine (TCM) preparation for treating stomachache, toothache, and rheumatoid arthritis. However, the active ingredients of ZN are not fully clarified due to its chemical complexity. In the present study, a double spectrum-effect analysis strategy was developed and applied to explore the bioactive components in herbs, and ZN was used as an example. Here, the chemical components in ZN were rapidly and comprehensively profiled based on the mass defect filtering-based structure classification (MDFSC) and diagnostic fragment-ion-based extension approaches. Furthermore, the fingerprints of 20 batches of ZN samples were analyzed by high-performance liquid chromatography, and the anti-inflammatory and antioxidant activities of the 20 batches of ZN samples were studied. Finally, the partial least squares regression (PLSR), gray relational analysis models, and Spearman's rank correlation coefficient (SRCC) were applied to discover the bioactive compounds in ZN. As a result, a total of 48 compounds were identified or tentatively characterized in ZN, including 35 alkaloids, seven coumarins, three phenolic acids, two flavonoids, and one lignan. The results achieved by three prediction models indicated that peaks 4, 12, and 17 were the potential anti-inflammatory compounds in ZN, whereas peaks 3, 5, 7, 12, and 13 were involved in the antioxidant activity. Among them, peaks 4, 5, 7, and 12 were identified as nitidine, chelerythrine, hesperidin, and oxynitidine by comparison with the standards and other references. The data in the current study achieved by double spectrum-effect analysis strategy had great importance to improve the quality standardization of ZN, and the method might be an efficiency tool for the discovery of active components in a complex system, such as TCMs.


Antibacterial activity of Pd(II) complexes with salicylaldehyde-amino acids Schiff bases ligands.
Palladium(II) complexes with Schiff bases ligands derived from salicylaldehyde and amino acids (Ala, Gly, Met, Ser, Val) have been synthesized and characterized by Fourier transform (FT)-IR, UV-Vis and (1)H-NMR spectroscopy. The electrospray mass spectrometry (ES-MS) spectrometry confirms the formation of palladium(II) complexes in 1/2 (M/L) molar ratio. All the Pd(II) complexes 1, [Pd(SalAla)2]Cl2; 2, [Pd(SalGly)2]Cl2; 3, [Pd(SalMet)2]Cl2; 4, [Pd(SalSer)2]Cl2; 5, [Pd(SalVal)2]Cl2; have shown antibacterial activity against Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia coli.


IS THERE A RELATIONSHIP BETWEEN COMPLICATIONS OF EARLY PREGNANCY AND BIOMETEOROLOGICAL FORECAST?
The aim of our study was to connect the possible complications of early pregnancy (miscarriage and symptomatic ectopic pregnancy) up to the 12th week of gestation with biometeorological conditions while assuming a greater number of incidents with an unfavorable biometeorological forecast. We performed a retrospective observational study using medical data of a single medical center of Department of Gynecology and Obstetrics, Sveti Duh University Hospital and meteorological data from the Croatian Meteorological and Hydrometeorological Service in Zagreb. We tracked the number of visits to the gynecology and obstetrics emergency unit on a daily basis during 2017. Days with five or more visits were selected and underwent further analysis, during which the number of miscarriages and symptomatic ectopic pregnancies was noted. The information from the biometeorological forecast was then extracted and added to the database. Our results did not show a statistically significant difference between the groups determined by biometeorological forecast in the number of spontaneous abortions or ectopic pregnancy. Also, statistically significant results did not follow the expected trend of the increasing number of complications related to worse biometeorological forecast, or vice versa, a decreased number of complications with better forecast. Our single-center retrospective analysis of emergency unit visits related to weather conditions did not show a connection between the complications of early pregnancy and biometeorological conditions. However, different results could emerge in future studies. Considering the large and high-quality database collected for this study, efforts in researching the connection between other gynecologic pathologies and weather conditions will be feasible.


Immunohistochemical study of the expression of epidermal growth factor receptor in benign prostatic hypertrophy, prostatic intraepithelial neoplasia and prostatic carcinoma.
The present study provides an analysis of immunohistochemical expression and localization of epidermal growth factor receptor (EGFR) in formalin fixed paraffin embedded specimens of prostate. Thirty-five cases each of benign prostatic hypertrophy (BPH) and prostatic carcinoma and 30 cases of prostatic intraepithelial neoplasia (PIN) were taken up for study. Streptavidin biotin peroxidase method was employed for immunohistochemical staining. EGFR positivity was observed in all the cases (100%) of BPH and PIN and in only 10 cases (28.5%) of prostatic carcinoma. In both BPH and PIN the basal cells revealed significantly higher intensity and percentage cell positivity than the luminal cells. Intensity and percentage of positively stained basal cells in BPH was higher than PIN basal cells but the difference was not statistically significant. The intensity and percentage cell positivity of BPH basal cells and PIN basal and luminal cells were significantly greater than the epithelial cells of prostatic carcinoma. Presently, the significance of variable expression of EGFR in various types of prostatic lesions is unknown.


Grafting of cyclopentadienyl ruthenium complexes on aminosilane linker modified mesoporous SBA-15 silicates.
Cyclopentadienyl ruthenium phosphane and carbene complexes are grafted on the surface of mesoporous SBA-15 molecular sieves through an aminosilane linker. The nature of the support after the grafting is examined by powder XRD, TEM and N(2) adsorption/desorption analysis. Elemental analysis, FT-IR, DRIFTS, TG-MS and MAS-NMR studies confirm the successful grafting of the complexes on the surface. The grafted materials are applied for catalytic aldehyde olefination and cyclopropanation.


Time-specific androgen blockade with flutamide inhibits testicular descent in the rat.
Inhibition of androgen action by flutamide, a nonsteroidal antiandrogen, blocked testicular descent in 40% of the testes exposed to this agent continuously from gestational day 13 through postpartal day 28. By contrast, only 11% of the testes failed to descend when blocked by 5 alpha-reductase inhibitors during the same period. Flutamide administration over narrower time intervals (gestational day 13-15, 16-17, or 18-19) revealed maximal interference with testicular descent after androgen inhibition during gestational days 16-17. No significant differences in testicular or epididymal weights were evident between descended and undescended testes; furthermore, no correlation was detected between the presence of epididymal abnormalities and testicular descent. These findings indicate that androgen inhibition during a brief period of embryonic development can block testicular descent. The mechanism through which this inhibition occurs remains to be elucidated.


Early malnourished rats are not affected by anorexia induced by a selective serotonin reuptake inhibitor in adult life.
The effect of early postnatal malnutrition upon food intake and its modulation by the selective serotonin reuptake inhibitor (SSRI) citalopram, was investigated in adult rats. Sixty four Wistar rats were allocated to two groups, according to their mother's diet during lactation. Mothers receiving a 23% protein diet fed the well-nourished group; mothers receiving 8% protein diet fed the malnourished. After weaning, all rats received the 23% protein diet ad libitum. On the 120th day after birth, each nutritional group was divided in two subgroups (each one, n = 16) which received a single daily injection of citalopram (10 mg/kg) or saline (0.9% NaCl) for 14 days. Chronic treatment with citalopram decreased both the food intake and weight gain in the well-nourished rats, but not in the malnourished ones. These data are consistent with findings concerning the nutritional manipulation of the nervous system during its higher vulnerable phase, suggesting that early malnutrition alters the effect of treatment of SSRI in adult rats, and that malnutrition during the critical period of brain development affects the serotoninergic system.


Stochastic modeling and numerical simulation of gene regulatory networks with protein bursting.
Gene expression is inherently stochastic. Advanced single-cell microscopy techniques together with mathematical models for single gene expression led to important insights in elucidating the sources of intrinsic noise in prokaryotic and eukaryotic cells. In addition to the finite size effects due to low copy numbers, translational bursting is a dominant source of stochasticity in cell scenarios involving few short lived mRNA transcripts with high translational efficiency (as is typically the case for prokaryotes), causing protein synthesis to occur in random bursts. In the context of gene regulation cascades, the Chemical Master Equation (CME) governing gene expression has in general no closed form solution, and the accurate stochastic simulation of the dynamics of complex gene regulatory networks is a major computational challenge. The CME associated to a single gene self regulatory motif has been previously approximated by a one dimensional time dependent partial integral differential equation (PIDE). However, to the best of our knowledge, multidimensional versions for such PIDE have not been developed yet. Here we propose a multidimensional PIDE model for regulatory networks involving multiple genes with self and cross regulations (in which genes can be regulated by different transcription factors) derived as the continuous counterpart of a CME with jump process. The model offers a reliable description of systems with translational bursting. In order to provide an efficient numerical solution, we develop a semilagrangian method to discretize the differential part of the PIDE, combined with a composed trapezoidal quadrature formula to approximate the integral term. We apply the model and numerical method to study sustained stochastic oscillations and the development of competence, a particular case of transient differentiation attained by certain bacterial cells under stress conditions. We found that the resulting probability distributions are distinguishable from those characteristic of other transient differentiation processes. In this way, they can be employed as markers or signatures that identify such phenomena from bacterial population experimental data, for instance. The computational efficiency of the semilagrangian method makes it suitable for purposes like model identification and parameter estimation from experimental data or, in combination with optimization routines, the design of gene regulatory networks under molecular noise.


Prevalence of Multiple Sclerosis in Rosario, Argentina.
The prevalence and incidence of Multiple Sclerosis have been increasing worldwide over the last decades. Most of the publications during the last years outstand the prevalence in Europe, North America and Latin America. In Argentina, data published on this topic is scarce and most of the studies took place in Buenos Aires. In a recent publication, we reported an MS prevalence rate of 30.3/100,000 in the overall extension of Santa Fe, Argentina, and in that study, we noticed that MS seemed to be more frequent in the southern region of the province. In this study we aimed to estimate the prevalence of MS in Rosario (the third most populated district of Argentina), placed in the southeastern part of Santa Fe province. This is a population based, cross-sectional study. We studied the members of the Province´s medical Care Program, a health maintenance organization of the public-sector employees that offers medical and health services to 567,819 members in the province 163,513 of whom have permanent residence in Rosario department, being a large representative sample. The selected prevalence date was 30 June 2019. Cases of MS were detected with a thorough search in the electronic databases of affiliates, linking and aggregating datasets. Seventy-nine MS cases were detected. The mean age was 47.7 (SD 13.5); female to male ratio was 3 to 1; the most frequent phenotype was relapsing-remitting (82.3%) followed by secondary-progressive (15.2%) and primary-progressive (2.5%). The crude prevalence rate of MS in Rosario as on 30 June 2019 was 48.3/100,000 inhabitants (95% CI: 48.28 - 48.35), 63.7/100,000 for females and 28.3/100,000 for males. The age-standardised MS prevalence rate was 43.4/100,000 inhabitants (95% CI:43.37 - 43.43) while the sex-standardised prevalence was estimated at 46.7/100,000 inhabitants (95% CI: 46.68 - 46.74). Making a proportional and linear projection to the total number of inhabitants, we deemed a total of 578 MS cases in Rosario, according to the 2010 census, and projected 620 cases by July 2019. This is the first study that provides epidemiological data of MS prevalence for Rosario. It confirms that there are differences in the distribution of MS in Santa Fe that favour the aggregation of cases in the southern part of the province. It also supplies updated and relevant data on the distribution of MS in Argentina and Latin America.


Psychiatric disorders and 15-month mortality in a community sample of older adults.
The relation between psychiatric disorders, defined by the Diagnostic Interview Schedule (DIS), and mortality over 15 months is compared in 3007 adults age 55 and over in the New Haven Epidemiologic Catchment Area (ECA) project. Our results indicate that the odds of dying are more than four times greater for individuals with affective disorders than for others in the sample, controlling for age, sex, and physical health. Cause of death is also examined. There were no suicides or deaths from external causes.


Intercellular interactions and cytokine responsiveness of peritoneal alpha/beta and gamma/delta T cells from Listeria-infected mice: synergistic effects of interleukin 1 and 7 on gamma/delta T cells.
Peritoneal gamma/delta T cells from Listeria-immune mice show an enhanced potential to expand when restimulated with antigens or mitogens in vitro (see companion paper [Skeen, M. J., and H. K. Ziegler. 1993. J. Exp. Med. 178:971]). When cocultured with peritoneal alpha/beta T cells, the gamma/delta T cell population expanded preferentially even when the in vitro stimulus was specific for the alpha/beta T cell population. Purified gamma/delta T cells did not respond to alpha/beta T cell-specific stimuli. If isolated T cell subsets were recombined in cell mixing experiments, the resulting proliferative response was greater than additive. Irradiated alpha/beta T cells could enhance the proliferation of responding gamma/delta T cells, but the effect was unidirectional; i.e., irradiated gamma/delta T cells did not stimulate responding gamma/delta T cells. This effect appeared to be cytokine mediated and did not require cell-cell contact. Both recombinant interleukin 2 (rIL-2) and rIL-7 could support the expansion of the gamma/delta T cells, while rIL-7 was only minimally stimulatory for the alpha/beta T cells. The magnitude of the response by gamma/delta T cells to rIL-7 exceeded the response to other in vitro stimuli, including immobilized anti-T cell receptor monoclonal antibody, and was 50-100-fold greater than the alpha/beta T cell response to IL-7. This unique sensitivity of gamma/delta T cells to IL-7 was strongly enhanced by the presence of accessory cells. These cells could be replaced by rIL-1, establishing a synergy for IL-1 and IL-7 as factors that could uniquely stimulate this gamma/delta T cell population. Isolated peritoneal gamma/delta T cells from Listeria-immune mice react to heat-killed Listeria preparations in the presence of macrophages accessory cells in a non-H-2-restricted manner. Considered collectively, these results suggest a potential mechanism by which gamma/delta T cells can predominate in epithelial tissues and at sites of infection.


Palliative intubation of malignant oesophageal strictures.
A 6-year experience (1981-1987) of palliative intubation of irresectable malignant oesophageal strictures is reported in 110 patients with a mean age of 70.3 (range 41-90) years. Pulsion intubation was performed on 71 patients, 11 (15.5%) of whom died, and traction intubation on 39 with 6 (15.4%) deaths. Seven deaths resulted from instrumental perforation, but six other patients survived perforation and left the hospital in satisfactory condition. Mean in-patient stay was 8 (range 1-26) days. Non-fatal tube-related complications were more common in pulsion intubation, but was found to be highly effective in relieving dysphagia, with shortened hospital stay (mean 6 days) and acceptable morbidity and mortality rates. These results indicate the trend towards, and the increased safety of, pulsion intubation.


Effects of the FIFA 11+ and a modified warm-up programme on injury prevention and performance improvement among youth male football players.
The effects of the FIFA11+ programme (the 11+) on ankle and groin injuries and performance have remained questionable. The latter, particularly, has potentially reduced the implementation rate and applicability of the programme. This study aimed to evaluate the mid-to-long-term effects of the 11+ and a modified programme including football-specific exercises on injury prevention and performance improvement. Three teams of the Iranian Youth League (division two) volunteered to participate in this study and were randomly assigned to two intervention groups (F11+; n = 29, M11+; n = 31) and a control group (n = 30). The F11+ followed the FIFA 11+ programme, whereas the M11+ performed modified exercises three times weekly as a warm-up protocol before training and competition through a football season. The control group carried out its routine warm-ups, including joggings, basic football drills, and static stretches, while having no injury prevention approaches. Lower extremity injuries, as well as exposure time for each player, were recorded. The football-specific performance was assessed using the Illinois Agility and Slalom Dribbling tests. ANOVA, Fisher Freeman Halton, and chi-square tests were used to analyze the data. Injury incidences differed significantly between groups (p = 0.02, C = 0.40), with M11+ reporting the lowest incidence. Significant differences between the pre- and posttest as well as differences between the groups for development over time were revealed for the Illinois agility and dribbling speed (p≤0.01). Both performance tests demonstrated a large time effect, as the effect sizes for time in agility and dribbling speed were 0.74 (CI = [0.66; 0.79]) and 0.86 (CI = [0.79; 0.87]), respectively. The effect size for the interaction can be categorized as medium, with 0.38 (CI = [0.25; 0.49]) for agility and 0.52 (CI = [0.40; 0.61]) for dribbling speed. M11+ showed the largest improvement in both. Mid-to-long-term application of a structured dynamic warm-up that integrates injury prevention and performance approaches may lower injury incidences and improve youth subelite players' performance. Although additional studies with larger samples are needed to prove the results of the current study, the amateur clubs/teams could integrate such twofold dynamic warm up into their routine training plan and benefit its advantages on injury prevention and performance improvement.


Purification, reconstitution, and subunit composition of a voltage-gated chloride channel from Torpedo electroplax.
The voltage-gated Cl- channel from Torpedo electroplax was purified in functional form by an immunoaffinity procedure. Channel activity was assayed by 36Cl- uptake into reconstituted liposomes and by direct recording after insertion into planar lipid bilayers. The purified channel displays the same "double-barreled" gating kinetics observed with native membranes, as well as the correct single-channel permeation characteristics. Preparations of active channels consist of a 90-kDa polypeptide, as expected from the known cDNA sequence. No associated subunits are present in the purified material. Direct protein sequencing confirms the absence of a cleavable signal sequence and demonstrates an N-terminus at Ser-2 of the cDNA-derived sequence. This "ClC-0" protein is lightly glycosylated, losing only approximately 2 kDa of sugar upon treatment with endoglycosidase H or N-glycanase. Most if not all of this glycosylation is found on Asn-365. This result necessitates revision of current transmembrane topology proposals, which have placed this residue on the cytoplasmic side of the membrane. Sedimentation in sucrose density gradients under activity-preserving conditions suggests the ClC-0 channel is slightly larger than the Na/K-ATPase alpha/beta-protomer (approximately equal to 150 kDa) and substantially smaller than the reduced form of the nicotinic acetylcholine receptor (approximately equal to 300 kDa). The detergent-solubilized ClC-0 channel, which invariably displays two Cl- diffusion pores in the active complex, is therefore built most likely as a homodimer of the 90-kDa protein purified here.


Postprandial lipaemia 10 and 34 hours after playing football: Does playing frequency affect the response?
Elevated postprandial triglyceride (TG) is associated with increased risk of cardiovascular disease. The time window for the last bout beneficial effect on postprandial lipaemia after football play is unknown. The aim of the present study was to examine whether playing affects postprandial TG during 1.5 day of reduced activity. Eighteen males were randomly allocated to perform either 1 (1FOOT; n = 9; age = 33.0 ± 5.0 yrs; body mass index = 24.2 ± 3.6 kg/m2) or 3 (3FOOT) consecutive days of 60-min 5 vs 5 football (n = 9; age = 32.8 ± 5.2 yrs; body mass index = 26.2 ± 4.1 kg/m2) matches across a 5-day study period. They arrived to the laboratory 10 hrs and 34 hrs after the final football session and blood samples were collected at fasted (0 min) and 45, 90, 240 and 360 min post a high fat load meal. There were non significant increase for postprandial TG AUC (9.1%; p = 0.17; 95%CI = -0.43 to 2.0; ES = -0.23) and iAUC (14.2%; p = 0.43; 95%CI = -0.92 to 1.9; ES = -0.24) between 10 and 34 hrs after the 1FOOT. For the 3FOOT, there was a non significant decrease in postprandial TG AUC (-2.7%; p = 0.73; 95%CI = -2.0 to 1.5; ES = 0.05) and iAUC (-17.5%; p = 0.41; 95%ci = -2.5 to 1.1; ES = 0.31) from 10 to 34 hrs, respectively. Performing three consecutive days of football exercise may offer no greater protective effect for postprandial TG before a period of reduced activity, compared to a single session.


Altered neuroendocrine regulation of luteinizing hormone secretion in postmenopausal women with Parkinson's disease.
The secretion of gonadotropins and the role exerted by the endogenous opioid system on luteinizing hormone (LH) secretion were investigated in 6 postmenopausal women affected by idiopathic Parkinson's disease (PD) as well as in 6 age- and weight-matched normal postmenopausal women as controls. The mean plasma follicle-stimulating hormone (FSH) and LH levels were evaluated both under basal conditions and after 20 days of conjugated estrogen administration (1.25 mg/day). At the same time, the activity of the endogenous opioid system was evaluated, as well as the LH response to the 4-hour infusion of the opioid antagonist naloxone (1.6 mg i.v. bolus followed by 1.6 mg/h). Both before and during estrogen administration, plasma FSH levels were similar in the two groups of subjects, whereas plasma LH levels were significantly lower (p less than 0.01) in parkinsonian than in control women. In each subject estrogen administration significantly blunted (p less than 0.01) plasma FSH levels. Plasma LH levels were reduced only in controls (p less than 0.05), but not in women with PD. In each subject, before estrogen administration, the plasma LH levels did not vary during naloxone infusion. In control women after 20 days of estrogen administration, the plasma LH levels significantly increased during naloxone infusion (p less than 0.01). By contrast, in women with PD, conjugated estrogens failed to restore the LH response to naloxone. The present results suggest that the neurotransmitter mechanisms, which regulate LH secretion, are altered, and, in particular, the activity of the endogenous opioid system is deficient in women with PD.


Basic science of the innate immune system and the lung.
Infections of the lung are prevented first by denial of access, mediated by humoral and cellular systems that obscure the pathogen's target, simultaneously acting to kill and clear the pathogen. Where pathogens penetrate these initial defence systems, rapid detection of their presence is achieved by their engagement with a large family of pattern recognition receptors, including Toll-like receptors, retinoic-acid-inducible gene-like helicases and Nod-like receptors. Activation of a coordinated inflammatory response is achieved by cooperative signalling between sentinel leucocytes such as alveolar macrophages and lung tissue, following which a profound pro-inflammatory response mobilizes phagocytes such as neutrophils and monocytes to the lung. Resolution of the innate immune response, largely dependent upon neutrophil apoptosis, results in restoration of normal tissue architecture. In chronic disease, dysregulated inflammation maintains these systems in a state of constant activation, potentially resulting in tissue damage and progressive disease.


Novel lectin-like bacteriocins of biocontrol strain Pseudomonas fluorescens Pf-5.
Bacteriocin LlpA, produced by Pseudomonas sp. strain BW11M1, is a peculiar antibacterial protein due to its homology to mannose-binding lectins mostly found in monocots (A. H. A. Parret, G. Schoofs, P. Proost, and R. De Mot, J. Bacteriol. 185:897-908, 2003). Biocontrol strain Pseudomonas fluorescens Pf-5 contains two llpA-like genes, named llpA1(Pf-5) and llpA2(Pf-5). Recombinant Escherichia coli cells expressing llpA1(Pf-5) or llpA2(Pf-5) acquired bacteriocin activity and secreted a 31-kDa protein cross-reacting with LlpA(BW11M1) antibodies. Antibacterial activity of the recombinant proteins was evidenced by gel overlay assays. Analysis of the antimicrobial spectrum indicated that LlpA1(Pf-5) and LlpA2(Pf-5) are able to inhibit P. fluorescens strains, as well as the related mushroom pathogen Pseudomonas tolaasii. LlpA-type bacteriocins are characterized by a domain structure consisting of tandem monocot mannose-binding lectin (MMBL) domains. Molecular phylogeny of these MMBL domains suggests that the individual MMBL domains within an LlpA protein have evolved separately toward a specific, as yet unknown, function or, alternatively, were acquired from different ancestral sources. Our observations are consistent with earlier observations, which hinted that MMBL-like bacteriocins represent a new family of antibacterial proteins, probably with a novel mode of action.


Effect of propionic acid on kinetics of acetate and oleate and on plasma and milk fatty acid composition of goats.
Entry rates of acetate and oleate and their incorporation into lipids of blood plasma and fatty acids of milk were studied in lactating goats fed a concentrate-roughage ration with propionic acid infused intraruminally at 0, 5.52, and 13.74 g/h by primed constant intravenous infusion of [1-carbon-14] acetate and [9, 10-hydrogen-3] oleate. Means for infusion rates were acetate, 60, 52, and 39 micrograms/ml blood plasma; propionate 9, 12, and 22 micrograms/ml; oleate, 19, 14, and 12 micrograms/ml; acetate entry rate, 3.9, 2.7, and 1.8 mmol/h per kg bodyweight; oleate entry rate, 47, 29, and 19 mumol/h per kg bodyweight; acetate oxidation rate, 2.0, 1.7, and 1.4 mmol/h per kg, and its contribution to the total carbon dioxide production, 16, 14, and 11%. Propionic acid increased incorporation of carbon-14 and hydrogen-3 into plasma lipids, elevated proportions of 7:0, 9:0, 11:0, 13:0, 15:0, and 17:0 fatty acids in milk, and tended to lower others. Specific radioactivities of milk fatty acids during infusion of propionic acid were elevated by 1.8 to 2.8 times, and total fatty acids in milk and plasma were lowered by 22 and 38%. Data support the glucogenic theory that propionic acid either directly or through gluconeogenesis stimulates insulin secretion, which in turn inhibits release of fatty acids from adipose tissue, resulting in milk fat depression.


The role of TCGA molecular classification in clear cell endometrial carcinoma.
Clear cell endometrial carcinoma (CCEC) represents a relatively rare and heterogeneous entity. Based on The Cancer Genome Atlas (TCGA) molecular classification, the risk stratification and management of endometrial cancer (EC) have been improved. Although the relationship of CCEC with the TCGA classification is less well understood, data has emerged to suggest that molecular classification plays an important role in the prognosis and management of CCEC. Most of patients with CCEC are characterized by p53abn or NSMP type and the prognosis of these patients is poor, whereas those with MMRd or POLEmut seem to have a favorable prognosis. Adjuvant therapy is recommended in CCEC with p53abn and NSMP. Advanced/recurrent CCEC with MMRd benefit much more from immune checkpoint inhibitors after the failure of platinum-based chemotherapy. In addition, bevacizumab plus chemotherapy upfront seems to improve outcomes of advanced/recurrent patients whose tumors harbored mutated TP53, including CCECs with p53abn. Further studies which exclusively recruit CCEC are urgently needed to better understand the role of molecular classification in CCEC. This review will provide an overview of our current understanding of TCGA classification in CCEC.


Comparison of Real-world Outcomes Between Patients Treated with Tapentadol ER or Oxycodone CR.
Background: The objective of this study was to compare health care utilization and costs between matched cohorts of chronic pain patients treated with the opioids tapentadol extended release (ER) or oxycodone controlled release (CR). Methods: This retrospective study used claims data from the Optum Research Database. Commercial and Medicare Advantage adult patients with ≥1 prescription fill for oxycodone CR or tapentadol ER between September 1, 2011 and September 30, 2012 were eligible. The date of the first observed oxycodone CR or tapentadol ER claim was the index date. Patients had continuous health plan enrollment for 6 months before and after the index date, ≥ 90 days supply of opioid therapy, and no index drug claims in the preindex period. Patients were propensity score matched in a 1:2 ratio (tapentadol ER : oxycodone CR). Results: The attributes of the matched cohorts (1,120 tapentadol ER and 2,240 oxycodone CR patients) appeared similar. In the 6 month post-index period, lower proportions of the tapentadol ER cohort than the oxycodone CR cohort had ≥1 inpatient stay (14.6% versus 20.5%; p<0.001) and ≥1 emergency department visit (33.4% versus 37.5%; p=0.021). The tapentadol ER compared with the oxycodone CR cohort had higher mean pharmacy costs ($4,263 versus $3,694; p <0.001), lower mean inpatient costs ($3,625 versus $6,309; p<0.001), and lower mean total healthcare costs ($16,510 versus $19,330; p=0.004). Conclusions: During follow-up, total mean healthcare costs were lower among tapentadol ER patients than oxycodone CR patients, and tapentadol ER patients were less likely to have an inpatient admission or emergency department visit.


Activity profile and physiological responses of Korean amateur football referees during matches.
[Purpose] The present study aimed to analyze and compare the activity profile and physiological responses of amateur football referees during competitive matches of high school and college students. [Subjects and Methods] Thirty referees (high school, 15; college, 15) were included in this study. The total distance covered, movement speed, and heart rate were measured using a global positioning system-enabled wireless heart rate monitor. The blood lactate concentration was measured immediately after the first and second half. [Results] College football referees covered a higher total distance than did their high school counterparts (7,547 m vs. 6,719 m). The maximal heart rate of college football referees was low in the first half alone, and the percentage of the heart rate within the "maximum" range was low throughout the game. [Conclusions] Refereeing imposes a significantly high physical load on the body while tracking player and ball movement. The present study suggests the need for developing and distributing physical training programs tailored for refereeing.


Angle noncritical phase-matched second-harmonic generation in the monoclinic crystal BaGa₄Se₇.
We performed a complete study of angle noncritical phase-matched second-harmonic generation in BaGa4Se7 in the mid-infrared region. We measured the dielectric frame orientation and showed that it does not rotate as a function of wavelength. We determined the phase-matching wavelengths and the associated spectral and angular acceptances along the x and y axes of the dielectric frame. We also estimated the magnitude of the nonlinear coefficients involved.


The response of HIV-associated lymphadenopathic Kaposi sarcoma to highly active antiretroviral therapy evaluated by 18F-FDG PET/CT.
The use of PET/CT for determining systemic Kaposi sarcoma (KS) involvement and treatment response remains to be formally assessed. Pretreatment human herpes virus-8 (HHV-8) viral load may predict clinical response of KS to therapy; however, its role in treatment monitoring and correlation with PET/CT findings is unknown. We report PET/CT findings and HHV-8 viral loads of a 46-year-old man with HIV-associated lymphadenopathic KS. After treatment with highly active antiretroviral therapy, an interval PET/CT showed partial response, despite an undetectable HHV-8 viral load. Chemotherapy was initiated, and the patient achieved a complete clinical, PET/CT, and virological response. PET/CT enabled monitoring of treatment response and led to changes in management.


Permissive role of Cdc2 activity induced from astrocytes in neurite outgrowth.
Following spinal cord injury, glial cells are recognized as major environmental factors hampering axon's regenerative responses. However, recent studies suggested that, in certain circumstances, reactive astrocytes may have a permissive role for axonal regeneration and functional recovery. Here, we report that Cdc2 activation in astrocytes is positively linked to axon growth. Cdc2 was strongly, but transiently, induced from reactive astrocytes within and around the injury cavity. Cdc2 levels in primary, non-neuronal cells prepared from injured spinal cord were up-regulated by extending the pre-injury period. Cdc2-mediated vimentin phosphorylation was strongly induced in astrocytes after long-term culture (7 days, LTC) as compared with short-term culture (3 days, STC). Induction levels of phospho-vimentin in LTC astrocytes were positively associated with increased neurite outgrowth in co-cultured dorsal root ganglion neurons. β3 integrin mRNA was induced in LTC astrocytes and activation of β3 integrin was regulated by Cdc2 activity. Furthermore, genetic depletion and pharmacological blockade experiments demonstrate that activation of Cdc2 and β3 integrin in LTC astrocytes is required for neurite outgrowth. Our data suggest that the Cdc2 pathway may play an important role in determining phenotypic expression of astrocytes such that astrocytes provide permissive environments for axonal regeneration following spinal cord injury.


A theoretical study on the mechanism of hydrogenation of carboxylic acids catalyzed by the Saito catalyst.
The mechanism of the ruthenium carboxylate-catalyzed hydrogenation of carboxylic acids was investigated by using density functional theory (DFT) calculations. The novel mechanism including two hydrogenation cycles was proposed for this reaction. The first cycle is the hydrogenation of the carboxylic acid to an aldehyde, while the second cycle is the hydrogenation of the aldehyde to an alcohol. These two catalytic cycles share similar elementary steps, including H2 heterolysis, hydride migration of the carboxylic acid or aldehyde, and catalyst regeneration. In this hydrogenation mechanism, the carboxylic acid is not only a reactant, but also an important proton source. Furthermore, the noncovalent interaction (e.g. hydrogen bonding interaction) between the ligand and carboxylic acid substrate could promote the hydrogenation of the carboxylic acid through stabilizing the transition state of the most energy-demanding step (i.e., hydride migration in the first catalytic cycle). Besides, the strong electron-donating ability of the dppb ligand could also facilitate the hydride migration.


Biological algorithm for data reconstruction.
An algorithm inspired by Genome sequencing is proposed which "reconstructs" a single long trajectory of a dynamical system from many short trajectories. This procedure is useful in situations when many data sets are available but each is insufficiently long to apply a meaningful analysis directly. The algorithm is applied to the Rössler and Lorenz dynamical systems as well as to experimental data taken from the Belousov-Zhabotinskii chemical reaction. Topological information was reliably extracted from each system and geometrical and dynamical measures were computed.


Salivary cortisol and psychosocial hazards at work.
Experimental and clinical evidence suggest that stress can lead to ill-health through the disregulation of the hypothalamic-pituitary-adrenal (HPA) axis. Studies to date have produced equivocal results likely due to different methodologies and failure to account for confounding factors. This investigation aimed to assess the relation between self-reported work-related stressors and salivary cortisol and to clarify the role of the potential confounders. Thirty-six call-handlers completed a self-administered job content questionnaire and collected seven daily salivary samples on two workdays and a weekend. The diurnal salivary cortisol output was expressed as cortisol awakening response (CAR), and cortisol output in the rest of the day. Salivary cortisol data were normalized by means of square root transformation. The generalized estimating equations method was used to assess the relation between job strain and cortisol levels after adjusting for gender, weekdays and adherence to the sampling schedule. Job strain significantly influenced the total amount of cortisol response to waking (high strain vs. low strain: 1.1 (0.3-2.0) nmol/L). The cortisol response to waking showed gender-specific differences [women excreting greater cortisol than men: 1.1 (0.3-1.9) nmol/L], and weekday differences [workdays vs. weekend: 1.0 (0.3-1.6) nmol/L]. Non-compliance with the sampling protocol was associated with lower salivary cortisol than in adherent subjects. Our results provide further evidence for the HPA axis involvement in the physiological response to work stress. The measure of the CAR showed to be the sensitive index to assess the physiological response to psychosocial factors. Gender, weekday, and protocol compliance were confounding factors.


Active surveillance for candidemia, Australia.
Population-based surveillance for candidemia in Australia from 2001 to 2004 identified 1,095 cases. Annual overall and hospital-specific incidences were 1.81/100,000 and 0.21/1,000 separations (completed admissions), respectively. Predisposing factors included malignancy (32.1%), indwelling vascular catheters (72.6%), use of antimicrobial agents (77%), and surgery (37.1%). Of 919 episodes, 81.5% were inpatient healthcare associated (IHCA), 11.6% were outpatient healthcare associated (OHCA), and 6.9% were community acquired (CA). Concomitant illnesses and risk factors were similar in IHCA and OHCA candidemia. IHCA candidemia was associated with sepsis at diagnosis (p<0.001), death <30 days after infection (p<0.001), and prolonged hospital admission (p<0.001). Non-Candida albicans species (52.7%) caused 60.5% of cases acquired outside hospitals and 49.9% of IHCA candidemia (p = 0.02). The 30-day death rate was 27.7% in those > or =65 years of age. Adult critical care stay, sepsis syndrome, and corticosteroid therapy were associated with the greatest risk for death. Systematic epidemiologic studies that use standardized definitions for IHCA, OHCA, and CA candidemia are indicated.


Tumor-infiltrating lymphocytes cultured in recombinant interleukin-2: enhancement of growth, cytotoxicity, and phenotypic expression of cytotoxic T-cell antigens by cyclophosphamide given intravenously prior to tumor harvest.
Tumor-infiltrating lymphocytes (TILs) have potent antitumor effects in murine models of advanced disease. Although these cells are 50-100 times more potent than lymphokine-activated killer cells against micrometastases, their antitumor benefits are virtually nonexistent against large tumor burdens unless cyclophosphamide is added to the immunotherapy regimen. In an effort to determine the effects of cyclophosphamide on TILs obtained from tumor-bearing animals, we harvested tumors from animals having received either 0, 50, or 100 mg/kg cyclophosphamide intravenously and investigated the expansion, cytotoxicity, and phenotypic expression of these TILs co-cultured in vitro with recombinant interleukin-2. TILs obtained from animals given cyclophosphamide, demonstrated a greater fold expansion than TILs obtained from normal animals (mean fold expansion on day 59 of culture: 85, 400, and 150 for TILs obtained from animals given 0, 50, and 100 mg/kg cyclophosphamide, n = 3 consecutive experiments). In addition, these TILs demonstrated enhanced cytotoxicity compared to controls [effector to target ratio 4:1, day 16 TILs, % lysis: 24, 35, and 45%, cyclophosphamide 0, 50, and 100 mg/kg, respectively, against the MCA-102 sarcoma, natural killer (NK) insensitive tumor; 29, 38, and 56% against the YAC-1 lymphoma, NK sensitive tumor]. Similar results were seen with day 34 and day 59 TILs. When phenotypic analysis was performed, TILs obtained from animals given cyclophosphamide consistently demonstrated a greater percent expression of the Thy1.2 and Lyt-2 antigens up to day 59 of culture when the experiments were terminated. The NK cell marker 49H.8 was expressed on the majority of TILs and its expression did not change with respect to the cyclophosphamide concentration. The increase in TIL number and cytotoxicity seen with cyclophosphamide given intravenously before tumor harvest could have important ramifications for human immunotherapy with TILs.


Conformer-Specific Desorption in Propanol Ices Probed by Chirped-Pulse Millimeter-Wave Rotational Spectroscopy.
We present a new technique for the detection of molecules desorbed from an ice surface using broad-band millimeter-wave rotational spectroscopy. The approach permits interrogation of molecules that have undergone the slow warmup process of temperature-programmed desorption (TPD), analogous to the warmup phase of icy grains in the interstellar medium as they approach the central protostar. The detection is conformer- and isomer-specific and quantitative, as afforded by chirped-pulse rotational spectroscopy. To achieve this, we combine ice TPD with buffer gas cooling, followed by detection in the millimeter-wave regime. In this report we examine the TPD profiles of n- and i-propanol, the former of which may be in five different conformational isomeric forms, and which display distinct desorption profiles. The limited conformational isomerization and temperature-dependent relative yields of n-propanol conformers observed show that the desorption is highly conformer-specific.


Takotsubo cardiomyopathy in myasthaenia gravis crisis confirmed by cardiac MRI.
Myasthaenia gravis crisis and Takotsubo cardiomyopathy are rare conditions that can be precipitated by emotional or physical stress. Myasthaenia gravis has a variety of cardiac manifestations but Takotsubo cardiomyopathy, particularly in male patients, has rarely been reported. We describe a unique case of a 70-year-old man who developed Takotsubo cardiomyopathy during his first presentation with a myasthaenia gravis crisis. He had not received plasmapharesis or immunoglobulin therapy. Striking ECG traces and cardiac MRI helped to confirm the diagnosis. Cardiac manifestations of myasthaenia gravis and myasthaenia gravis itself have overlapping symptoms; the importance of cardiac monitoring and clinical vigilance in such cases is discussed. The utility of cardiac MRI in assessing cardiac manifestations of myasthaenia gravis is also highlighted.


The Future of Family Medicine version 2.0: reflections from Pisacano scholars.
The Future of Family Medicine (FFM) project has helped shape and direct the evolution of primary care medicine over the past decade. Pisacano Scholars, a group of leaders in family medicine supported by the American Board of Family Medicine, gathered for a 2-day symposium in April 2013 to explore the history of the FFM project and outline a vision for the next phase of this work-FFM version 2.0 (v2.0). After learning about the original FFM project (FFM v1.0), the group held interactive discussions using the World Café approach to conversational leadership. This commentary summarizes the discussions and highlights major themes relevant to FFM v2.0 identified by the group. The group endorsed the FFM v1.0 recommendations as still relevant and marvelled at the progress made toward achieving many of those goals. Most elements of FFM v1.0 have moved forward, and some have been incorporated into policy blueprints for reform. Now is the time to refocus attention on facets of FFM v1.0 not yet realized and to identify key aspects missing from FFM v1.0. The Pisacano Scholars are committed to moving the FFM goals forward and hope that this expression of the group's vision will help to do so.


Primary immune response to Helix pomatia haemocyanin in malignant melanoma. Relationship between 19S and 7S antibody response and in vitro lymphocyte transformation.
The primary immune response to Helix pamatia haemocyanin (HPH) was investigated in sixty-one patients with various clinical signs of malignant melanoma and compared to that of controls. Anti-HPH antibody response was only found abnormal in patients with widespread disease and visceral metastases, apparent from decreased 7S and increased 19S antibody levels. In vitro HPH-induced lymphocyte transformation, in contrast, was not only decreased in this late stage, but also in part of the patients in the beginning of the disease and was then correlated with subsequent tumour recurrence within 6 months. Generally, patients with positive lymphocyte transformation showed significantly higher 7S anti-HPH titres (mean 3-3 +/- 2-4 than patients without (1-7 +/- 1-6). In contrast to the lymphocyte transformation reaction, anti-HPH antibody response was not correlated with subsequent tumour recurrence. The same phenomenon of decreased 7S and increased 19S antibody response as in the melanoma patients with far advanced disease was observed to a less extent in controls with ageing. Anamnestic antibody responses to diphtheria and tetanus toxoid and serum IgG and IgA levels were found normal. Elevated serum IgM levels were more frequently observed in patients with localized (19/31) and disseminated disease (11/29) than in controls (2/31). These results suggest that melanoma patients develop impaired T-lymphocyte functions in final stages with visceral metastases. A previous study showed that minor defects in lymphocyte function in an early stage as measured by HPH-induced lymphocyte transformation have predictive value for subsequent tumour recurrence.


[Increased chance of dying among nursing home patients with lower body weight].
To investigate a possible relationship between changes in weight and mortality among nursing home patients of 65 years and older, after adjustment for age, sex and main diagnosis. Descriptive longitudinal study. Dutch nursing home in Delft, the Netherlands. For three years after admission the body weight of patients newly admitted from April 1st 1987 to April 1st 1989 was measured every three months under the same conditions. It was recorded if a patient died or was discharged in this three-year period. The effects of body weight at admission, sex, age and diagnostic category (psychogeriatric, neurological, orthopaedic, oncological, other) on mortality were analysed using the univariate Cox regression analysis. The effect of the time-dependent covariate body weight on mortality in the nursing home was analysed with a combined logistic regression analysis per 3-month interval. The mean weight of the females increased from 59.7 kg (n = 194) at the time of admission to 63.7 kg (n = 57) three years later and that of the males from 64.7 kg (n = 70) to 70.3 (n = 11) due to the low body weight of those who died. During the first 3 months after admission mortality was high (14%) and only diagnosis related. In subsequent 3-month periods there was a significant association between mortality and weight, weight change, age and sex: the risk of mortality was higher for those with a lower body weight, weight loss, higher age and male sex. Nursing home patients with a low body weight or loss of weight after the first 3 months after admission, have an increased risk of death.


Callogorgia lucaya sp. nov., a new species of deep-sea Primnoidae (Anthozoa: Octocorallia) from the western Atlantic.
As part of an ongoing reassessment of the genus Callogorgia, we describe C. lucaya sp. nov. from the western Atlantic, with a distribution so far restricted to the Straits of Florida and northern Caribbean. The species was partially described in an unfinished manuscript of Dr. Frederick Bayer and differs from all other western Atlantic species by commonly lacking outer-lateral body wall scales, having a small number of abaxials, and having body wall sclerites that are externally almost smooth. (Species Zoobank LSID: urn:lsid:zoobank.org:act:CF8F92FC-8F8A-4A8C-AF4A-52E12B58F7B1).


Adolescents Who Vape Nicotine and Their Experiences Vaping: A Qualitative Study.
Understanding adolescent perceptions of vaping and roles it plays in their lives is needed to design effective interventions to help adolescents quit. We explored vaping experiences of 11 adolescents from initiation through quit attempts. A convenience sample of students who vaped in the last 90 days was recruited from one suburban high school in Massachusetts. Qualitative interviews were transcribed and coded. An inductive thematic analysis approach was employed. Areas of agreement and range of responses in code reports were summarized. Eleven open-ended semi-structured interviews were conducted (mean = 32.5 minutes each). Vaping initiation often occurred when socializing with friends who also supplied vaping devices. Vaping was "something to do" and new flavors engaged adolescents further. Solitary activities coupled with vaping included video gaming, getting ready for school, talking on the phone, or studying. Peak hours for vaping included morning, before and after school, before parents returned from work, and after parents went to bed. Several vaped to address anxiety/stress. For some, anxiety control was a main reason for vaping. Participants were concerned about health effects and nicotine dependence. Cost and health effects were drivers of quitting. Quit strategies relied on willpower and distraction. Peers have powerful influences on the initiation and maintenance of adolescent vaping. Vaping habits can become routinized into adolescent lives. Addiction is a concern although nicotine's anxiolytic effects were valued by many. Social connection was enhanced by communal vaping, sharing, and common vernacular, secrecy and rule-breaking. We describe the context in which adolescents vape nicotine, their reasons for vaping, and reasons to quit. This information can inform the development of interventions to better address adolescents' triggers to vape, and social and psychosocial barriers to quitting. Our findings suggest a desire to quit vaping but a limited awareness of quitting strategies.


Pharmacokinetics of miconazole in serum and exudate of pelvic retroperitoneal space after radical hysterectomy and pelvic lymphadenectomy.
Due to the increased number of compromised hosts with fungal infections, doctors have recently started prescribing antifungal agents. In the field of gynecology, however, the choice of which drug to use has been difficult. The efficacies of these drugs depend on their antifungal spectra, potencies and concentrations in tissues. The present study was designed to investigate the pharmacokinetics of miconazole in the exudate of the retroperitoneal space that is formed after radical hysterectomy and pelvic lymphadenectomy. A total of 600 mg of miconazole was administered to the patients for exactly 60 min using an automatic drip-infusion pump. The parameters of the formulas analyzed by the two-compartment model were determined using the least-squares method, and a simulation curve was made. The maximum drug concentration (Cmax) of miconazole in serum was 6.26 mg/l 1 h after drip infusion commencement and the t1/2 in serum was 8.86 h. The value of the area under the time-serum concentration curve (AUC) in serum was 19.13 mg/h per l. The Cmax of miconazole in the exudate of the retroperitoneal space was 0.13 mg/l 2.48 h after the drip infusion was started. The value of AUC in the exudate was 2.52 mg/h per l.


Multilocus genotypic characterization of Escherichia coli O157:H7 recovered from food sources.
Escherichia coli O157:H7 strains (n = 33) recovered from different food sources in Egypt were characterized using molecular assays to identify strain genotypes associated with various levels of pathogenic potential. Genotypic characterization included: lineage-specific polymorphism assay (LSPA-6), Shiga-toxin-encoding bacteriophage insertion site assay (SBI), clade 8 typing, Tir (A255 T) polymorphism, and variant analysis of Shiga toxin 2 gene (Stx 2a and Stx 2c), and anti-terminator Q genes (Q 933 and Q 21). Genotypes LI/II (76%), SBI 1 (60·6%), clade 8 (69·7%), Tir (255 T) (72·7%) and Stx 2c (45·5%) were found to be significantly more frequent compared to other genetic markers in the strains analysed. Multivariable analysis revealed a significant association between LPSA-6 and clade types as well as Tir (A255 T). To the best of our knowledge, this is the first study to report the characterization of these genetic markers in E. coli O157:H7 strains in the Middle East and Africa.


Simulated Microgravity Disrupts Cytoskeleton Organization and Increases Apoptosis of Rat Neural Crest Stem Cells Via Upregulating CXCR4 Expression and RhoA-ROCK1-p38 MAPK-p53 Signaling.
Neural crest stem cells (NCSCs) are a population of multipotent stem cells that are distributed broadly in many tissues and organs and are capable of differentiating into a variety of cell types that are dispersed throughout three germ layers. We are interested in studying the effects of simulated microgravity on the survival and self-renewal of NCSCs. NCSCs extracted from the hair follicle bulge region of the rat whisker pad were cultured in vitro, respectively, in a 2D adherent environment and a 3D suspension environment using the rotatory cell culture system (RCCS) to simulate microgravity. We found that rat NCSCs (rNCSCs) cultured in the RCCS for 24 h showed disrupted organization of filamentous actin, increased globular actin level, formation of plasma membrane blebbing and neurite-like artifact, as well as decreased levels of cortactin and vimentin. Interestingly, ∼70% of RCCS-cultured rNCSCs co-expressed cleaved (active) caspase-3 and neuronal markers microtubule-associated protein 2 (MAP2) and Tuj1 instead of NCSC markers, suggesting stress-induced formation of neurite-like artifact in rNCSCs. In addition, rNCSCs showed increased C-X-C chemokine receptor 4 (CXCR4) expression, RhoA GTPase activation, Rho-associated kinase 1 (ROCK1) and p38 mitogen-activated protein kinase (MAPK) phosphorylation, and p53 expression in the nucleus. Incubation of rNCSCs with the Gα protein inhibitor pertussis toxin or CXCR4 siRNA during RCCS-culturing prevented cytoskeleton disorganization and plasma membrane blebbing, and it suppressed apoptosis of rNCSCs. Taken together, we demonstrate for the first time that simulated microgravity disrupts cytoskeleton organization and increases apoptosis of rNCSCs via upregulating CXCR4 expression and the RhoA-ROCK1-p38 MAPK-p53 signaling pathway.


New insights into the biological roles of immune cells in neural stem cells in post-traumatic injury of the central nervous system.
Traumatic injuries in the central nervous system, such as traumatic brain injury and spinal cord injury, are associated with tissue inflammation and the infiltration of immune cells, which simultaneously affect the self-renewal and differentiation of neural stem cells. However, the tissue repair process instigated by endogenous neural stem cells is incapable of restoring central nervous system injuries without external intervention. Recently, resident/peripheral immune cells have been demonstrated to exert significant effects on neural stem cells. Thus, the restoration of traumatic injuries in the central nervous system by the immune intervention in neural stem cells represents a potential therapeutic method. In this review, we discuss the roles and possible mechanisms of immune cells on the self-renewal and differentiation of neural stem cells along with the prognosis of central nervous system injuries based on immune intervention. Finally, we discuss remaining research challenges that need to be considered in the future. Further elucidation of these challenges will facilitate the successful application of neural stem cells in central nervous system injuries.


Systemic activation of NLRP3 inflammasome in patients with severe primary Sjögren's syndrome fueled by inflammagenic DNA accumulations.
Sjögren's syndrome (SS) patients manifest high cell-free DNA (cf-DNA) levels in serum, associated with impaired DNaseI activity. Undegraded DNA may accumulate in tissues and act as an inflammasome-activating signal. Herein, we investigated the occurrence of aberrant DNA build-up in various biologic compartments of SS patients and its correlation with the activity of NLRP3 and AIM2 inflammasomes. For this purpose, we evaluated sera, PBMC, circulating monocytes and salivary glands (SG) from different SS patient subgroups and controls. We found that SS patients at high risk for lymphoma and those with established lymphoma display high serum cf-DNA levels, substantial extranuclear DNA accumulations in PBMC and SG tissues, a unique NLRP3 inflammasome gene signature in PBMC, and significantly increased serum IL-18 and ASC levels. In these patients, the circulating monocytes manifested NLRP3 inflammasome activation and increased response to NLRP3 stimuli, whereas SG-infiltrating macrophages exhibited signs of NLRP3 activation and pyroptosis. Cell-free nucleic acids isolated from patients' sera competently primed the activation of both NLRP3 and AIM2 inflammasomes in healthy monocytes. SS patients also manifested diminished DNaseI activity in serum and DNaseII expression in PBMC, which inversely correlated with indices of inflammasome activation. DNaseII gene-silencing in healthy monocytes led to cytoplasmic DNA deposition and activation of inflammasome-related genes and of caspase1. Our data reveal the occurrence of systemic NLRP3 inflammasome activation in severe SS, which is associated with widespread extranuclear accumulations of inflammagenic DNA and impaired DNA degradation. These findings can provide novel biomarkers and new therapeutic targets for the management of SS patients with adverse outcomes.


Phase variation of pili of Corynebacterium pilosum.
Whether or not (?) phase variation occurs in the pili of Corynebacterium pilosum was biologically examined using the colony enzyme-linked immunosorbent assay (ELISA) blot test with anti-pili immune serum. From the densely piliated clone (35P+) of C. pilosum 35, non-piliated variants were isolated at a frequency of 2.45 x 10(-3). From one of the non-piliated variants (designated as P(-11)/35P+), a piliated variant was isolated at a frequency of 4.68 x 10(-4), one log less frequently than the non-piliated variant. From this piliated variant as designated (P+4/P(-11)/35P+), a non-piliated variant was isolated at a frequency of 3.86 x 10(-3). C. pilosum was thus alternated between piliated and non-piliated at a fairly high frequency, suggesting that the pili may undergo phase variation. This is the first finding of phase variation of the pili of Gram-positive bacteria.


Aquaporin 1 Is an Independent Marker of Poor Prognosis in Lung Adenocarcinoma.
Aquaporin 1 (AQP1) overexpression has been shown to be associated with uncontrolled cell replication, invasion, migration, and tumor metastasis. We aimed to evaluate AQP1 expression in lung adenocarcinomas and to examine its association with clinicopathological features and prognostic significance. We also investigated the association between AQP1 overexpression and epithelial-mesenchymal transition (EMT) markers. We examined AQP1 expression in 505 cases of surgically resected lung adenocarcinomas acquired at the Seoul National University Bundang Hospital from 2003 to 2012. Expression of AQP1 and EMT-related markers, including Ecadherin and vimentin, were analyzed by immunohistochemistry and tissue microarray. AQP1 overexpression was associated with several aggressive pathological parameters, including venous invasion, lymphatic invasion, and tumor recurrence. AQP1 overexpression tended to be associated with higher histological grade, advanced pathological stage, and anaplastic lymphoma kinase (ALK) translocation; however, these differences were not statistically significant. In addition, AQP1 overexpression positively correlated with loss of E-cadherin expression and acquired expression of vimentin. Lung adenocarcinoma patients with AQP1 overexpression showed shorter progression-free survival (PFS, 46.1 months vs. 56.2 months) compared to patients without AQP1 overexpression. Multivariate analysis confirmed that AQP1 overexpression was significantly associated with shorter PFS (hazard ratio, 1.429; 95% confidence interval, 1.033 to 1.977; p=.031). AQP1 overexpression was thereby concluded to be an independent factor of poor prognosis associated with shorter PFS in lung adenocarcinoma. These results suggested that AQP1 overexpression might be considered as a prognostic biomarker of lung adenocarcinoma.


Term delivery in a woman with severe congenital neutropenia, treated with growth colony stimulating factor.
The patient was diagnosed in childhood as having severe congenital neutropenia and had recurrent admissions with severe infections. In 1987, prior to getting married, she was sterilized. She continued to require i.v. antibiotics when she contracted a severe infection. On one occasion, she was treated with growth colony stimulating factor (G-CSF). Her increased neutrophil count was sustained following this treatment. In June 1993, she wished to start a family and underwent in-vitro fertilization (IVF) treatment. G-CSF was given prior to oocyte retrieval. She conceived on her first cycle and an ultrasound scan revealed a singleton pregnancy. Throughout the course of the pregnancy, her white cell count was monitored closely and remained at <1.0x10(9)/l. The pregnancy progressed uneventfully and at 37 weeks gestation she was admitted for G-CSF injections. At 38 weeks she was delivered of a boy weighing 3350 g, by elective Caesarean section. His white cell count was normal. This is the first case of G-CSF being used before conception and during pregnancy in a patient with congenital neutropenia. It shows that advances in cytokine therapy and close interdisciplinary liaison can lead to a successful outcome and help patients, who would otherwise remain childless, to achieve a family.


Healthcare Ethics Consultation in Austria: Joining the International Path of Professionalization.
AbstractHealthcare ethics consultation has been developed, practiced, and analyzed internationally. However, only a few professional standards have evolved globally in this field that would be comparable to standards in other areas of healthcare. This article cannot compensate for this situation. It contributes to the ongoing debate on professionalization by presenting experiences with ethics consultation in Austria, though. After exploring its contexts and providing an overview of one of its primary ethics programs, the article analyzes the underlying assumptions of "ethics consultation" as an essential effort on the path to professionalize ethics consultation.


Human mitochondria in health, disease, ageing and cancer.
In every human cell there are hundreds of mitochondria, which are required for oxidative phosphorylation as well as many other metabolic processes. Each mitochondrion contains approximately 5 mitochondrial DNA molecules. These circular DNAs of 16.5 kb in size contain only 39 genes. Mutations in mitochondrial DNA are responsible for many diseases. Alterations in these molecules may also play a role in ageing and in tumour formation.


Adherence of Treponema pallidum subsp. pallidum in the rabbit placenta.
Congenital syphilis is the consequence of transplacental passage of Treponema pallidum. A system was developed to deliver virulent T. pallidum, Nichols strain, through an isolated uterine horn of a pregnant rabbit in order to investigate the mechanism by which T. pallidum is able to cross the placenta. While the pregnant rabbit was anesthetized, the ovarian artery and the uterine vein were cannulated and attached to a peristaltic pump. Treponema pallidum (2-5 x 10(8) in 10-15 ml RPMI-1640) were circulated via the peristaltic pump throughout the horn for 2 hr, after which the placentas were removed, fixed in formalin, and embedded in paraffin. This system was used to investigate treponemal binding to rabbit placenta at Day 20 and 26 during the gestation period of the rabbit (29-32 days). Examination of 5-microns Dieterle silver stained tissue sections revealed (i) a greater number of spirochetes in the later gestational stage placentas (Day 26) than in the earlier placentas (Day 20), (ii) organisms adhering to the trophoblastic tissue surrounding the maternal blood channels, and (iii) organisms appearing to be in the process of penetrating the trophoblastic tissue or that had completely penetrated from the channels into the trophoblastic elements. We suggest that T. pallidum may be adhering to placental components that are differentially expressed during gestation of the rabbit.


Genotypic variability based association identifies novel non-additive loci DHCR7 and IRF4 in sero-negative rheumatoid arthritis.
Sero-negative rheumatoid arthritis (RA) is a highly heterogeneous disorder with only a few additive loci identified to date. We report a genotypic variability-based genome-wide association study (vGWAS) of six cohorts of sero-negative RA recruited in Europe and the US that were genotyped with the Immunochip. A two-stage approach was used: (1) a mixed model to partition dichotomous phenotypes into an additive component and non-additive residuals on the liability scale and (2) the Levene's test to assess equality of the residual variances across genotype groups. The vGWAS identified rs2852853 (P = 1.3e-08, DHCR7) and rs62389423 (P = 1.8e-05, near IRF4) in addition to two previously identified loci (HLA-DQB1 and ANKRD55), which were all statistically validated using cross validation. DHCR7 encodes an enzyme important in cutaneous synthesis of vitamin D and DHCR7 mutations are believed to be important for early humans to adapt to Northern Europe where residents have reduced ultraviolet-B exposure and tend to have light skin color. IRF4 is a key locus responsible for skin color, with a vitamin D receptor-binding interval. These vGWAS results together suggest that vitamin D deficiency is potentially causal of sero-negative RA and provide new insights into the pathogenesis of the disorder.


Cytokine profile of broncho-alveolar lavage in BOOP and UIP.
The aim of our study was to compare clinical and BAL features of patients with bronchiolitis obliterans organizing pneumonia (BOOP) with those of patients with usual interstitial pneumonia (UIP) and control subjects. This study reports on 14 patients with idiopathic BOOP. Diagnosis was made upon histology. Lung function tests were mostly normal. Chest X-ray and CT showed always a patchy consolidation, often associated with ground glass pattern. BAL was performed for cytology and for ELISA assessment of several cytokines (IL8, ILI0, IL12, gamma-interferon, IL 18, monocyte chemoattractant protein- 1). Cytology of BAL in BOOP showed a pattern of lymphocytic alveolitis (Lymphocytes: 0.36 x 10(6)/ml) associated with an increase in neutrophil and eosinophil counts (0.13 and 0.04 x 10(6)/ml respectively). Mean BALf levels in pg/ml of MCP-1, IL12 and IL18 were significantly increased in BOOP with respect to controls and UIP patients, while in UIP patients only a significant increase of IL8, MCP-1 and IL18 with respect to controls was detected. In addition, BALf levels of IL10, an anti-inflammatory cytokine, were significantly higher in BOOP patients with respect to controls and UIP patients. These findings are consistent with a marked degree of macrophage and lymphocyte activation in BOOP with an expansion of T helper-1 response. The concomitant increase of IL10 could be related to a limitation of the inflammatory process and the fibrotic evolution typical of this clinical picture.


Population Differentiation at the PVT1 Gene Locus: Implications for Prostate Cancer.
Genetic variation in susceptibility to complex diseases, such as cancer, is well-established. Enrichment of disease associated alleles in specific populations could have implications for disease incidence and prevalence. Prostate cancer (PCa) is a disease with well-established higher incidence, prevalence, and worse outcomes among men of African ancestry in comparison to other populations. PCa is a multi-factorial, complex disease, but the exact mechanisms for its development and progression are unclear. The gene desert located on chromosome 8q24 is associated with aggressiveness of PCa. Interestingly, the non-protein coding gene locus Plasmacytoma Variant Translocation (PVT1) is present at chromosome 8q24 and is overexpressed in PCa. PVT1 gives rise to multiple transcripts with potentially different molecular and cellular functions. In an analysis of the PVT1 locus using data from the 1000 Genomes Project, we found the chromosomal region spanning PVT1 exons 4A and 4B to be highly differentiated between African and non-African populations. We further investigated levels of gene expression of PVT1 exons 4A and 4B and observed significant overexpression of these exons in PCa tissues relative to benign prostatic hyperplasia and to normal prostate tissues obtained from men of African ancestry. These results indicate that PVT1 exons 4A and 4B may have clinical implications in PCa a conclusion supported by the observation that transient and stable overexpression of PVT1 exons 4A and 4B significantly induce greater prostate epithelial cell migration and proliferation. We anticipate that further exploration of the role of PVT1 exons 4A and 4B may lead to the development of diagnostic, therapeutic, and other clinical applications in PCa.


Iron nanoparticle-induced activation of plasma membrane H(+)-ATPase promotes stomatal opening in Arabidopsis thaliana.
Engineered nanomaterials (ENMs) enable the control and exploration of intermolecular interactions inside microscopic systems, but the potential environmental impacts of their inevitable release remain largely unknown. Plants exposed to ENMs display effects, such as increase in biomass and chlorophyll, distinct from those induced by exposure to their bulk counterparts, but few studies have addressed the mechanisms underlying such physiological results. The current investigation found that exposure of Arabidopsis thaliana to nano zerovalent iron (nZVI) triggered high plasma membrane H(+)-ATPase activity. The increase in activity caused a decrease in apoplastic pH, an increase in leaf area, and also wider stomatal aperture. Analysis of gene expression indicated that the levels of the H(+)-ATPase isoform responsible for stomatal opening, AHA2, were 5-fold higher in plants exposed to nZVI than in unexposed control plants. This is the first study to show that nZVI enhances stomatal opening by inducing the activation of plasma membrane H(+)-ATPase, leading to the possibility of increased CO2 uptake.


A theoretical study of the mechanism of selective fluorination of saturated hydrocarbons by molecular fluorine. Participation of CHCl3 solvent molecules in the ionic process.
The fluorination reaction of methane and isobutane by molecular F(2) with and without CHCl(3) solvent was studied with the ab initio and ONIOM theoretical calculations. The electrophilic pathway for the RH + F(2) reaction in general is a two-step process, consisting of hydride abstraction leading to an intermediate of the type R(+)(delta)...HF...F(-)(delta), followed by complicated rearrangement to give the electrophilic substitution product, RF + HF. In the case of methane, the overall barrier for this reaction is too high for the reaction to take place under mild conditions even in the presence of CHCl(3) solvent molecules. In the case of isobutane without CHCl(3), the electrophilic pathway has a high rate-determining barrier of 25 kcal/mol, and is not likely to take place; the radical process forming t-Bu* + HF + F* may be preferred. However, the t-Bu(+)(delta)...HF...F(-)(delta) intermediate and, in particular, the transition state TS2 for rearrangement of the intermediate are highly ionic, and are stabilized dramatically when a few CHCl(3) solvent molecules form a solvation cage. The electrophilic reaction for isobutane + F(2) has a low overall barrier when at least three CHCl(3) molecules are present and can take place under mild conditions with full retention of configuration.


Urinary and bowel management in cloacal exstrophy: A long-term multi-institutional cross-sectional study.
We sought to evaluate long-term surgical urinary and bowel management in cloacal exstrophy (CE) in a multi-institutional study. We performed a cross-sectional study of people with CE and covered variants managed at five participating institutions. Those with <1 year follow-up or born with variants without hindgut involvement were excluded. Primary outcomes were methods of urinary and bowel management. Urinary management included: voiding via urethra, clean intermittent catheterizations (CIC), incontinent diversion and incontinent in diaper. Bowel management included: intestinal diversion (colostomy/ileostomy) and pull-through (with/without MACE). We evaluated three age groups: children (<10 years), older children (10 to <18) and adults (≥18). We assessed if management varied by age, institution or time (born≤2000 vs. >2000). A total of 160 patients were included (40% male). Median follow-up was 15.2 years (36% children, 22% older children, 43% adults). While 42% of children were incontinent in diapers, 73% of older children and adults managed their bladder with CIC, followed by incontinent urinary diversion (21%) (p < 0.001, Table). CIC typically occurred after augmentation (88%) via a catheterizable channel (89%). Among older children and adults, 86% did not evacuate urine per urethra and 28% of adults had an incontinent urinary diversion. No child or adult voided per urethra. Age-adjusted odds of undergoing incontinent diversion was no different between institutions (p = 0.31) or based on birthyear (p = 0.08). Most patients (79%) had an intestinal diversion, irrespective of age (p = 0.99). Remaining patients had a pull-through, half with a MACE. The probability of undergoing bowel diversion varied significantly between institutions (range: 55-91%, p = 0.001), but not birth year (p = 0.85). We believe this large long-term data presents a sobering but realistic view of outcomes in CE. A limitation is our data does not assess comorbidities or patient-reported outcomes. Rarity of volitional urethral voiding in CE forces the question of whether is a potentially unachievable goal. We advocate thoughtful surgical decision making and thorough counseling about appropriate expectations, distinguishing between volitional voiding and urinary and fecal dryness. In this long-term, multi-institutional study of patients with CE, 94% of older children and adults manage their bladder with incontinent diversion or CIC. Nearly 80% of patients, regardless of age, have an intestinal diversion. Given that no patients were dry and voided via urethra and 86% of older patients do not evacuate urine per urethra, these data bring into question what functional goals are achievable when performing reconstructive surgery for these patients.


Genome-Wide Transcriptome Analysis Reveals Extensive Alternative Splicing Events in the Protoscoleces of Echinococcus granulosus and Echinococcus multilocularis.
Alternative splicing (AS), as one of the most important topics in the post-genomic era, has been extensively studied in numerous organisms. However, little is known about the prevalence and characteristics of AS in Echinococcus species, which can cause significant health problems to humans and domestic animals. Based on high-throughput RNA-sequencing data, we performed a genome-wide survey of AS in two major pathogens of echinococcosis-Echinococcus granulosus and Echinococcus multilocularis. Our study revealed that the prevalence and characteristics of AS in protoscoleces of the two parasites were generally consistent with each other. A total of 6,826 AS events from 3,774 E. granulosus genes and 6,644 AS events from 3,611 E. multilocularis genes were identified in protoscolex transcriptomes, indicating that 33-36% of genes were subject to AS in the two parasites. Strikingly, intron retention instead of exon skipping was the predominant type of AS in Echinococcus species. Moreover, analysis of the Kyoto Encyclopedia of Genes and Genomes pathway indicated that genes that underwent AS events were significantly enriched in multiple pathways mainly related to metabolism (e.g., purine, fatty acid, galactose, and glycerolipid metabolism), signal transduction (e.g., Jak-STAT, VEGF, Notch, and GnRH signaling pathways), and genetic information processing (e.g., RNA transport and mRNA surveillance pathways). The landscape of AS obtained in this study will not only facilitate future investigations on transcriptome complexity and AS regulation during the life cycle of Echinococcus species, but also provide an invaluable resource for future functional and evolutionary studies of AS in platyhelminth parasites.


Cytochemical Localization of Polysaccharides in Dendrobium officinale and the Involvement of DoCSLA6 in the Synthesis of Mannan Polysaccharides.
Dendrobium officinale is a precious traditional Chinese medicinal plant because of its abundant polysaccharides found in stems. We determined the composition of water-soluble polysaccharides and starch content in D. officinale stems. The extracted water-soluble polysaccharide content was as high as 35% (w/w). Analysis of the composition of monosaccharides showed that the water-soluble polysaccharides were dominated by mannose, to a lesser extent glucose, and a small amount of galactose, in a molar ratio of 223:48:1. Although starch was also found, its content was less than 10%. This result indicated that the major polysaccharides in D. officinale stems were non-starch polysaccharides, which might be mannan polysaccharides. The polysaccharides formed granules and were stored in plastids similar to starch grains, were localized in D. officinale stems by semi-thin and ultrathin sections. CELLULOSE SYNTHASE-LIKE A (CSLA) family members encode mannan synthases that catalyze the formation of mannan polysaccharides. To determine whether the CSLA gene from D. officinale was responsible for the synthesis of mannan polysaccharides, 35S:DoCSLA6 transgenic lines were generated and characterized. Our results suggest that the CSLA family genes from D. officinale play an important role in the biosynthesis of mannan polysaccharides.


Mid-term changes in blood metal ion levels after Articular Surface Replacement arthroplasty of the hip.
Our first aim was to determine whether there are significant changes in the level of metal ions in the blood at mid-term follow-up, in patients with an Articular Surface Replacement (ASR) arthroplasty. Secondly, we sought to identify risk factors for any increases. The study involved 435 patients who underwent unilateral, metal-on-metal (MoM) hip resurfacing (HRA) or total hip arthroplasty (THA). These patients all had one measurement of the level of metal ions in the blood before seven years had passed post-operatively (early evaluation) and one after seven years had passed post-operatively (mid-term evaluation). Changes in ion levels were tested using a Wilcoxon signed-rank test. We identified subgroups at the highest risk of increase using a multivariable linear logistic regression model. There were significant increases in the levels of metal ions for patients who underwent both MoM HRA (Chromium (Cr): 0.5 parts per billion (ppb); Cobalt (Co): 1.1 ppb) and MoM THA (Cr: 0.5 ppb; Co: 0.7 ppb). In a multivariable model considering MoM HRAs, the change in the levels of metal ions was influenced by female gender (Co: Odds Ratio (OR) 1.42; p = 0.002 and Cr: OR 1.08; p = 0.006). The change was found to be irrespective of the initial level for the MoM HRAs, whereas there was a negative relationship between the initial level and the change in the level for those with a MoM THA (Co: OR -0.43; p < 0.001 and Cr: OR -0.14; p = 0.033). The levels of metal ions in the blood increase significantly over the period until mid-term follow-up in patients with both a MoM HRA and those with a MoM THA. We recommend that the levels of metal ions be measured most frequently for women with a MoM HRA. While those with a MoM THA appear to stabilise at a certain level, the accuracy of this trend is not yet clear. Vigilant follow-up is still recommended. Cite this article: Bone Joint J 2017;99-B(4 Supple B):33-40.


Molecular characterization of novel reciprocal translocation t(6;14) in an Epstein-Barr virus-transformed B cell precursor.
An in vitro culture of FLEB14 cells, an Epstein-Barr virus-transformed B cell precursor containing the germ line immunoglobulin genes, gave rise to a uniclonally expanded variant, FLEB14 delta 3, which was rearranged at the immunoglobulin heavy-chain gene locus. Cytogenetic analysis showed that FLEB14 delta 3 had a novel reciprocal translocation, t(6;14)(q15;q32). Molecular cloning of the rearranged DNA fragments and determination of their nucleotide sequence revealed that the recombination event was reciprocal, imprecise, and nonhomologous and took place in the S mu region, like those found in Burkitt's lymphoma cells. We propose a molecular model to explain this genetic event which may be relevant to class switch recombination. The translocated sequence of chromosome 6 did not contain any known oncogenes, although the sequence is conserved among mammals. FLEB14 delta 3 did not show tumorigenicity.