Machine learning-based CT texture analysis to predict HPV status in oropharyngeal squamous cell carcinoma: comparison of 2D and 3D segmentation.
To compare the CT texture feature reproducibility of 2D and 3D segmentations and their machine learning (ML)-based classifications for predicting human papilloma virus (HPV) status in oropharyngeal squamous cell carcinoma (OPSCC). Data about 47 patients with pathological OPSCC (15 HPV positive and 32 HPV negative) were collected from a public database. Using 2D and 3D manual segmentations, 1032 texture features were extracted from contrast-enhanced CT images. Intraclass correlation coefficients (ICCs) were calculated to evaluate intraobserver and interobserver reproducibility. Collinearity analysis and a wrapper-based subset search algorithm were used for feature selection. Models were created using k-nearest neighbors (k-NN), logistic regression (LR), and random forest (RF) alone and with a synthetic minority oversampling technique (SMOTE). Classifier performance was assessed using 10-fold cross-validation. Compared with 2D segmentation (468 of 1032, 45.3%), 3D segmentation (576 of 1032, 55.8%) yielded more texture features with reliable reproducibility (good to excellent in both intraobserver and interobserver analyses) (p < 0.001). RF and k-NN classifiers failed to achieve better classification performance using 3D features than using 2D features either alone or with SMOTE. The best models for 2D and 3D segmentations were both created using RF, which alone achieved areas under the curve (AUCs) of 0.880 and 0.847, respectively, and with SMOTE, AUCs of 0.953 and 0.920, respectively, were achieved. Three-dimensional segmentation had better CT texture feature reproducibility, but 2D segmentation showed better performance. Considering the cost, 2D segmentation is more recommended for ML-based classification of HPV status of OPSCC. • Three-dimensional segmentation had better CT texture feature reproducibility than 2D segmentation. • Despite yielding more features with reliable reproducibility, 3D segmentation failed to provide better classification performance as compared to 2D for predicting HPV status of oropharyngeal squamous cell carcinoma. • The best models for 2D and 3D segmentations were both created using random forest classifier.


Programs for Preventing Depression in Adolescence: Who Benefits and Who Does Not? An Introduction to the Supplemental Issue.
We introduce this supplemental issue of Prevention Science, which brings together a set of papers from leading investigators who have conducted trials testing whether intervention programs prevent adolescent depression. Using data from these trials, these papers explore a series of factors that might account for variation in intervention benefit, employing several novel methods for assessing effect heterogeneity. These studies follow two general paradigms: three papers report findings from single randomized preventive intervention trials, while the remaining papers develop and apply new methods for combining data from multiple studies to evaluate effect heterogeneity more broadly. Colleagues from NIMH and SAMHSA also provide commentaries on these studies. They conclude that synthesis of findings from multiple trials holds great promise for advancing the field, and progress will be accelerated if collaborative data sharing becomes the norm rather than the exception.


Aortic valve ochronosis: a rare manifestation of alkaptonuria.
Alkaptonuric ochronosis is a heritable disorder of tyrosine metabolism, with various systemic abnormalities related to pigment deposition and degeneration of collagen and other tissues, including the heart and aorta. A 65-year-old woman with alkaptonuric ochronosis and a history of four joint replacements required aortic valve replacement for severe aortic stenosis. Operative findings included ochronosis of a partly calcified aortic valve and the aortic intima. The aortic valve was removed at surgery and histologically investigated. Light microscopic examination of the aortic valve revealed intracellular and extracellular deposits of ochronotic pigment and a chronic inflammatory infiltrate. Beside the case representation, the disease history, aetiology, pathogenesis, clinical presentation and treatment of aortic valve ochronosis are reviewed.


Neurochemical development of the hippocampal region in the fetal rhesus monkey. I. Early appearance of peptides, calcium-binding proteins, DARPP-32, and monoamine innervation in the entorhinal cortex during the first half of gestation (E47 to E90).
Although the entorhinal cortex is a key structure connecting the hippocampal formation with the rest of the cerebral cortex, little is known about its early chemoanatomical development in primates. In the present study, a cytoarchitectonic analysis and immunocytochemical detection of somatostatin, neurotensin, parvalbumin, calbindin-D 28K, DARPP-32, as well as tyrosine hydroxylase, dopamine-beta-hydroxylase, and serotonin, were carried out on serial sections of the entorhinal cortex of six rhesus monkey fetuses aged E47 to E90 (gestation period 165 days). At E56 the cortical plate of the entorhinal cortex already exhibited a sublamination; at E64 the lamina dissecans was partly formed, allowing the emergence of the lamina principalis externa and interna, and at E83 most of the regional and laminar subdivisions characteristic of the adult cortex could be identified, except for the rhinal sulcus restricted to a small dimple. The neurochemical development paralleled the early cytoarchitectonic differentiation, both largely preceding that of the neighboring cortical areas. The somatostatin-like immunoreactive innervation, first detected at E56, was very dense as early as E64 and displayed by E83 a laminar distribution similar to that found in the adult. Labeled neurons indicated an intrinsic origin for this innervation but an extrinsic connection might be present as labeled fibers in the subplate of the entorhinal cortex were in continuity with positive fibers in the intermediate zone of the hippocampal formation. A faint neurotensin-like immunoreactivity first detected at E64 became prominent at E83 in the entorhinal cortex but stopped abruptly at the anlage of the rhinal sulcus. The lack of neurotensin-labeled neurons contrasted with their presence in other parts of the hippocampal region and suggested a precocious extrinsic connection. Only rare parvalbumin-LIR neurons were detected at midgestation, whereas calbindin-D 28K was expressed from E47 on in Cajal-Retzius cells and from E56 on in various types of neurons in the cortical plate and subplate. Most characteristic was a category of medium-sized, deeply stained calbindin-LIR neurons, present only in the lamina principalis externa and possibly corresponding to the population of large neurons described by Kostovic et al. (1990, Soc Neurosci Abstr 16:846) in early developing entorhinal cortex of human fetuses. These and probably other neurons were also DARPP-32-positive, suggesting the possibility of an early dopaminergic regulation. Indeed, the monoaminergic innervation of the entorhinal cortex was detected from E56 on and gradually increased in density, displaying areal and laminar differences in the distribution of the dopaminergic, noradrenergic, and serotoninergic afferents.(ABSTRACT TRUNCATED AT 400 WORDS)


Local structures of fluid with discrete spherical potential: Theory and grand canonical ensemble Monte Carlo simulation.
Grand canonical Monte Carlo simulation and theoretical calculations based on Ornstein-Zernike (OZ) integral equation and third order+second order perturbation density functional theory (DFT) are performed to study a system of spherical particles interacting through a core-softened (CS) potential combining a repulsive square soft core and an attractive square well. Both theoretical predictions and simulation results reveal peculiar homogeneous and inhomogeneous local structures originating from the discontinuous nature of the CS potential. The bulk radial distribution function displays discontinuities at the distances coinciding with the ranges of the successive repulsive and attractive parts in the CS potential function. The density profiles of confined CS fluid show the shapes arising from the complex interplay among the steric effects and the competition between the repulsive and attractive parts of the CS potential. Satisfactory agreement between the theoretical results and simulation data leads to the following conclusions: (i) a modified hypernetted chain approximation combined with a hard sphere bridge function, which has been recently proposed by one of the authors of this study, is sufficiently reliable for the structural studies of CS fluid, and (ii) the third order+second order perturbation DFT, which has proven successful for the study of inhomogeneous structure of model fluids with continuous intermolecular potential function, posses a high adaptability to be applied for various types of interaction potentials and performs well also in the case of discontinuous CS model.


Comparative distribution and validity of DSM-IV and DSM-5 diagnoses of eating disorders in adolescents from the community.
DSM-5 changes for eating disorders (EDs) aimed to reduce preponderance of non-specified cases and increase validity of specific diagnoses. The objectives were to estimate the combined effect of changes on prevalence of EDs in adolescents and examine validity of diagnostic groupings. A total of 3043 adolescents (1254 boys and 1789 girls, Mage = 14.19 years, SD = 1.61) completed self-report questionnaires including the Eating Disorder Diagnostic Scale. Prevalence of full-threshold EDs increased from 1.8% (DSM-IV) to 3.7% (DSM-5), with a higher prevalence of bulimia nervosa (1.6%) and the addition of the diagnosis of purging disorder (1.4%); prevalence of binge eating disorder was unchanged (0.5%), and non-specified cases decreased from 5.1% (DSM-IV) to 3.4% (DSM-5). Validation analyses demonstrated that DSM-5 ED subgroups better captured variance in psychopathology than DSM-IV subgroups. Findings extend results from previous prevalence and validation studies into the adolescent age range. Improved diagnostic categories should facilitate identification of EDs and indicate targeted treatments.


Siderophore typing, a powerful tool for the identification of fluorescent and nonfluorescent pseudomonads.
A total of 301 strains of fluorescent pseudomonads previously characterized by conventional phenotypic and/or genomic taxonomic methods were analyzed through siderotyping, i.e., by the isoelectrophoretic characterization of their main siderophores and pyoverdines and determination of the pyoverdine-mediated iron uptake specificity of the strains. As a general rule, strains within a well-circumscribed taxonomic group, namely the species Pseudomonas brassicacearum, Pseudomonas fuscovaginae, Pseudomonas jessenii, Pseudomonas mandelii, Pseudomonas monteilii, "Pseudomonas mosselii," "Pseudomonas palleronii," Pseudomonas rhodesiae, "Pseudomonas salomonii," Pseudomonas syringae, Pseudomonas thivervalensis, Pseudomonas tolaasii, and Pseudomonas veronii and the genomospecies FP1, FP2, and FP3 produced an identical pyoverdine which, in addition, was characteristic of the group, since it was structurally different from the pyoverdines produced by the other groups. In contrast, 28 strains belonging to the notoriously heterogeneous Pseudomonas fluorescens species were characterized by great heterogeneity at the pyoverdine level. The study of 23 partially characterized phenotypic clusters demonstrated that siderotyping is very useful in suggesting correlations between clusters and well-defined species and in detecting misclassified individual strains, as verified by DNA-DNA hybridization. The usefulness of siderotyping as a determinative tool was extended to the nonfluorescent species Pseudomonas corrugata, Pseudomonas frederiksbergensis, Pseudomonas graminis, and Pseudomonas plecoglossicida, which were seen to have an identical species-specific siderophore system and thus were easily differentiated from one another. Thus, the fast, accurate, and easy-to-perform siderotyping method compares favorably with the usual phenotypic and genomic methods presently necessary for accurate identification of pseudomonads at the species level.


Studies of glycemia and glucosuria in diabetics after breakfast meals of different composition.
In series I, The effect of four isocaloric, ordinary diabetic breakfast meals on the blood glucose level and on the urinary glucose loss was tested in 9 adult diabetes and three healthy subjects (ages 60 to 75) as well as in five insulin-treated diabetics (ages 18 to 40). All breakfasts derived 20% of the energy from protein, 35% from fat, and 45% from carbohydrates (CHO). The oligosaccharides amounted to 5, 30, 45, and 65% of CHO, respectively. In series II, four isocaloric breakfast meals with CHO amounting to 35 or 70% of the energy, oligosaccharides furnishing one-third or two-thirds of the CHO at each level, were tested in the same way in eight of adult diabetics and in the three healthy subjects. The two breakfasts with the highest percentage of oligosaccharides in series I caused the smallest increase in the blood glucose level in the adult subjects. In the adults as well as in the juveniles, these breakfasts gave rise to clear symptoms of hypoglycemia and/or a feeling of a deficient satiety in several subjects. In juveniles, the initial blood glucose level turned out to be of greater importance for the outcome of the test than the kind of carbohydrates consumed. In series II, The CHO-rich meals caused significantly greater blood glucose increases than did the low-CHO meals. Despite the unexpected findings that ordinary diabetics breakfast meals, high in oligosaccharides, gave a smaller increase in blood glucose than did meals in which wheat starch predominated, the use of diets rich in oligosaccharides should be avoided because of the risk of overconsumption owing to hypoglycemic symptoms or to the deficient feeling of satiety.


The Dacron ligament prosthesis in anterior cruciate ligament reconstruction. A four-year review.
We studied 40 patients who underwent reconstruction for chronic anterior cruciate ligament deficiency with a Dacron ligament prosthesis using a modified MacIntosh over-the-top technique, augmented with iliotibial band. Thirty patients had undergone at least 1 prior surgical procedure on the affected knee, but only 7 patients had previous anterior cruciate ligament reconstruction. All patients were followed for a mean of 47.5 months. The results at final followup demonstrated an average side-to-side arthrometer difference of 1.0 mm. The Lysholm score improved from 65 preoperatively to 89 at the end of the review; the Tegner activity level score improved from 3 to 5. Objectively, 75% of the patients had a negative Lachman test result and 95.1% of the subjects had negative or trace pivot shift results at review. Mild knee pain was still present with day-to-day activity in 87.7% of the patients. Complications occurred in 27.5% of patients, including five who had implant ruptures and two who had their grafts removed. Synovitis was a significant problem. Based on our failure criteria, 47.5% (19) of the subjects had failed results. In this study, radiologic evidence of tracer separation greater than 1 cm was a criterion of failure. With inclusion of tracer separation, the failure rate increased to 60.0% (24). Multiple previous surgeries of any type had an adverse effect on results. Damage to secondary stabilizers in these cases increased failure rate. Based on the high complication and failure rates, and relatively poor end result in this retrospective review, we cannot recommend this procedure.


Central and peripheral catecholamine synthesizing enzymes during the development of two-kidney, one clip hypertension in rats.
The activities of the catecholamine synthesizing enzymes have been determined in discrete brain areas and in peripheral tissues of rats, at different times after clipping the left renal artery (two-kidney Goldblatt hypertension, 2KGH) and in sham operated animals. Three days after clipping the only enzymatic change was a slight decrease in plasma dopamine-beta-hydroxylase (DBH) activity. Ten days after clipping no change in enzymatic activity was found at the central level. However, the DBH and the phenylethanolamine-N-methyltransferase (PNMT) activities were increased in the adrenal medulla (+49.0%, P less than 0.001 and +36.6%, P less than 0.001, respectively) and DBH activity was also increased in the superior cervical ganglia (+22.8%, P less than 0.01). These data suggest that sympathetic hyperactivity is present in 2KGH rats when hypertension is established. In addition, as this type of hypertension does not alter the PNMT activity in brainstem areas, it seems that the alterations in PNMT activity reported for genetically hypertensive rats are unlikely to be secondary to the elevated blood pressure.


Aqueous Nanoscale Systems.
In the past five years the Laboratory for fundamental BioPhotonics (LBP) has worked on developing new technology that can access the molecular structure and nanoscale properties of buried aqueous interfaces and aqueous solutions. Using these methods a better understanding of the important role that water plays in (nanoscale and interfacial) processes can be obtained. These processes include the long-range interaction of ions with water, structural and charge anomalies of the hydrophobic/aqueous interface, the formation and stabilization of amphiphilic aqueous droplet interfaces, the formation and molecular properties of the electric double layer, as well as membrane structure and hydration. The result of our work on these themes is summarized for this special issue article.


Common Blepharitis Related to Phthiriasis Palpebrarum: Argon Laser Phototherapy.
A 42-year-old woman was admitted to Mersin University, Department of Ophthalmology Clinic with itching and burning sensation of the right eye for 3 weeks. In her slit-lamp examination, nits and lice, attached to the upper and lower eyelashes of her right eye, were observed. Lice and nits were destroyed by argon laser phototherapy and were removed with the help of a fine forceps thereafter. Argon laser phototherapy is a quick, effective, and safe treatment modality for phthiriasis palpebrarum.


Segmental organisation of the head in the embryo of Drosophila melanogaster : A blastoderm fate map of the cuticle structures of the larval head.
Embryos of Drosophila melanogaster were irradiated in the presumptive head region with a UV-laser microbeam of 20 μm diameter at two developmental stages, the cellular blastoderm and the extended germ band. The ensuing defects were scored in the cuticle pattern of the head of the first-instar larva, which is described in detail in this paper. The defects caused by irradiating germ band embryos when morphologically recognisable lobes appear in the head region were used to establish the segmental origin of various head structures. This information enabled us to translate the spatial distribution of blastoderm defects into a fate map of segment anlagen. The gnathal segments derive from a region of the blastoderm between 60% and 70% egg length (EL) dorsally and 60% and 80% ventrally. The area anterior to the mandibular anlage and posterior to the stomodaeum is occupied by the small anlagen of the intercalary and antennal segments ventrally and dorsally, respectively. The labrum, which originates from a paired anlage dorsally at 90% EL, is separated from the remaining head segments by an area for which we did not observe cuticle defects following blastoderm irradiation, presumably because those cells give rise to the brain. The dorsal and lateral parts of the cephalo-pharyngeal skeleton appear to be the only cuticle derivatives of the non-segmental acron. These structures derive from a dorso-lateral area just behind the putative brain anlage and may overlap the latter. In addition to the segment anlagen, the regions of the presumptive dorsal pouch, anterior lobe and post-oral epithelium, whose morphogenetic movements during head involution result in the characteristic acephalic appearance of the larva, have been projected onto the blastoderm fate map. The results suggest that initially the head of the Drosophila embryo does not differ substantially from the generalised insect head as judged by comparison of fate map and segmental organisation.


Effect of the solvent on recognition properties of molecularly imprinted polymer specific for ochratoxin A.
A molecularly imprinted polymer specific for the mycotoxin ochratoxin A has been synthesised using a non-covalent approach. The polymer has shown an excellent affinity and specificity for the target template in aqueous solutions. The binding experiments, NMR study and molecular modelling have proven that the template recognition by polymer originates from the shape complementarity of binding sites. The binding mechanism is critically depended on factors that affect the polymer conformation. Thus the variation in buffer concentration, pH and presence of organic solvent, which affect the polymer swelling or shrinking, had a profound effect on the polymer recognition properties.


Extreme Conductance Suppression in Molecular Siloxanes.
Single-molecule conductance studies have traditionally focused on creating highly conducting molecular wires. However, progress in nanoscale electronics demands insulators just as it needs conductors. Here we describe the single-molecule length-dependent conductance properties of the classic silicon dioxide insulator. We synthesize molecular wires consisting of Si-O repeat units and measure their conductance through the scanning tunneling microscope-based break-junction method. These molecules yield conductance lower than alkanes of the same length and the largest length-dependent conductance decay of any molecular systems measured to date. We calculate single-molecule junction transmission and the complex band structure of the infinite 1D material for siloxane, in comparison with silane and alkane, and show that the large conductance decay is intrinsic to the nature of the Si-O bond. This work highlights the potential for siloxanes to function as molecular insulators in electronics.


Neonatal 6-hydroxydopamine and adult SKF 38393 treatments alter dopamine D1 receptor mRNA levels: absence of other neurochemical associations with the enhanced behavioral responses of lesioned rats.
To study potential biochemical correlates of dopamine (DA) and serotonin receptor supersensitivity, rats were lesioned at 3 days after birth with 6-hydroxydopamine (6-OHDA; 67 micrograms in each lateral ventricle; desipramine pretreatment, 20 mg/kg i.p., 1 h) and then sensitized with the DA D1 agonist, SKF 38393 HCl (3.0 mg/kg i.p. per day) either ontogenetically (daily, for 28 consecutive days from birth) and/or in adulthood (four weekly injections, 6-9 weeks from birth). Controls received vehicle in place of 6-OHDA or SKF 38393. Enhanced locomotor responses were observed after SKF 38393 at 6 weeks, only in rats that received SKF 38393 + 6-OHDA in ontogeny. Locomotor responses were further enhanced in this group after the last of four weekly SKF 38393 injections at the 9th week. These weekly SKF 38393 treatments also produced enhanced responses in 6-OHDA rats that did not receive SKF 38393 in ontogeny. When striata were studied at 11 weeks, the percentages of high and low affinity DA D1 binding sites were not altered. Basal as well as DA-, NaF-, and forskolin-stimulated adenylyl cyclase activities also were not changed. Dot blot analysis showed that there was a reduction of mRNA levels for DA D1, but not serotonin1C, receptors in the 6-OHDA groups. However, SKF 38393 at 6-9 weeks eliminated this alteration. Based on these findings it can be proposed that supersensitization may be a consequence of altered neuronal cross talk rather than an imbalance of receptor elements per se.


Heterokaryosis and alkaloid production in Claviceps purpurea.
Strain 275 FI of Claviceps purpurea, which produces large amounts of peptide alkaloids in submerged culture, is a heterokaryon; after several generations on agar media, it segregates the single components. These components (by us labeled V, C, and W) are stable and produce almost no alkaloids under described conditions of submerged culture. The mycelium of strain 275 FI consists of hyphae with multinucleate cells and does not produce conidia. Strains V, C, and W form numerous anastomoses when grown together on agar. By combining strains V and C, a heterokaryon similar to 275 FI in appearance has been obtained. This new strain produces amounts of alkaloids much larger than those produced by V and C separately or in associated submerged culture. We conclude, therefore, that in strain 275 FI the heterokaryotic condition is favorable to the production of alkaloids. Several conidiaproducing cultures of C. purpurea of various origins, as well as sclerotia of the same species, have been examined. The results demonstrated that the heterokaryotic condition is rare in the cultures, but it is frequent in the mycelium from sclerotia. Since it is known that the production of alkaloids is typical of the sclerotial phase in C. purpurea, it is suggested that this capacity is related to the heterokaryosis of the producing strains.


Impact of source doping on the performance of vertical InAs/InGaAsSb/GaSb nanowire tunneling field-effect transistors.
In this paper, we analyze experimental data from state-of-the-art vertical InAs/InGaAsSb/GaSb nanowire tunneling field-effect transistors (TFETs) to study the influence of source doping on their performance. Overall, the doping level impacts both the off-state and on-state performance of these devices. Separation of the doping from the heterostructure improved the subthreshold swing of the devices. The best devices reached a point subthreshold swing of 30 mV/dec at 100 x higher currents than previous Si-based TFETs. However, separation of doping from the heterostructure had a significant impact on the on-state performance of these devices due to effects related to source depletion. An increase in the doping level helped to improve the on-state performance, which also increased the subthreshold swing. Thus, further optimization of doping incorporation with the heterostructure will help to improve vertical InAs/InGaAsSb/GaSb nanowire TFETs.


Cardiac expression and subcellular localization of the p38 mitogen-activated protein kinase member, stress-activated protein kinase-3 (SAPK3).
Despite the interest in the roles that mitogen-activated protein kinases (MAPKs) play in the heart, the role of the different MAPK isoforms has been relatively poorly defined. A third isoform of p38 MAPK, known variously as stress-activated protein kinase-3 (SAPK3), p38- gamma or ERK6, has been previously shown to differ from p38- alpha/ beta both in its molecular weight and its lack of inhibition by the compound SB203580. We have generated monoclonal antibodies with specificity for SAPK3 demonstrated by immunoblot analysis, immunofluorescence studies, and cloning of SAPK3 from a rat heart cDNA expression library. By immunoblotting, we confirmed high expression of SAPK3 in fast, slow and mixed fibre types of murine skeletal muscle and observed significant expression restricted to heart, lung, thymus and testes. In addition to expression in normal heart (human, mouse, rat, dog and pig), we observed constant expression in diseased human heart, as well as control and hypertrophic cultured neonatal rat cardiac myocytes. Immunolocalization in cultured cardiac myocytes followed by confocal microscopy showed punctate, non-nuclear SAPK3 staining. In contrast, p38- alpha/ beta staining was non-punctate and distributed throughout the cytosol and nucleus. Whereas treatment with Leptomycin B to prevent nuclear export processes promoted higher levels of p38- alpha/ beta staining in cardiac myocyte nuclei, there was no apparent change in SAPK3 localization under these conditions. These differences between p38- alpha/ beta and SAPK3 probably reflect the specialized functions of SAPK3 and emphasize the need to evaluate SAPK3 upstream activators and downstream targets in the heart.


[The history of cardiology in Vienna].
Established by Auenbrugger the physical diagnostic increased the special occupation with diseases of the heart. The "anatomical clinic" in Vienna developed it to a before anticipated exactitude. Later Nothnagel and Wenckebach payed special attention to the symptom of angina pectoris. Austrian pioneers of electrocardiography were Rothberger and Winterberg.


Self-supporting carbon glass films: a PAS standard sample?
The preparation of self-supporting carbon glass films is described. Their spectral and photoacoustic parameters are presented indicating that these films are an optimum choice as standard samples for photoacoustic spectroscopy with gas-coupled microphones.


Rapid detection of toxigenic Clostridium difficile in fecal samples by magnetic immuno PCR assay.
Rapid detection of toxigenic Clostridium difficile in fecal samples was accomplished with the magnetic immuno PCR assay (MIPA). Elaborate DNA extraction techniques were unnecessary. First, we generated a mouse monoclonal antibody (MAb) reactive with only C. difficile, Clostridium sordellii, and Clostridium bifermentans. Then, magnetic beads were coated with the MAb, incubated with fecal samples to allow binding with C. difficile, extracted from the stool with a magnet, and processed in the PCR with primers specific for the toxin B gene. After optimizing MIPA by raising the number of PCR cycles from 35 to 40 and adding Chelex 100 to the PCR mixture, we found a sensitivity of 96.7%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 94.1% when compared with the culture of cytotoxic C. difficile from fecal samples. MIPA is a rapid, easy, and sensitive PCR method for demonstrating the presence of toxigenic C. difficile in stool samples and avoids the disadvantage of elaborate extraction of DNA from fecal samples.


The effect of 4-aminopyridine on the frequency-force relationship of isolated left atria of guinea-pigs.
The effect of 4-aminopyridine (4-AP) on the frequency-force relationship of isolated electrically-driven left atria of guinea-pigs was studied. 4-AP increased atrial contractility between the optimum and pessimum frequencies, but not the rested state contractions. The compound increased the time to peak force and the relaxation time. Results obtained indicate that 4-AP-induced positive inotropic effect is different from that induced by noradrenaline, and supports the view that the compound exerts a direct action on the myocardium to increase contractility. Release of endogenous noradrenaline by 4-AP cannot, however, be excluded.


[Combined spinal epidural anesthesia for emergency cesarean section].
We performed a retrospective analysis to evaluate the usefulness of combined spinal epidural anesthesia (CSEA) in emergency cesarean section compared with conventional spinal anesthesia. A retrospective chart analysis of patients who had undergone emergency cesarean section over a 7 year period was performed. We extracted at random 150 patients planned by spinal anesthesia, and 150 patients planned by separate-needle CSEA. Patient's data were gathered and analyzed according to the anesthetic technique used. There were no significant differences between the two groups in age, weight, and BMI. Although the time to incision and the time to delivery were significantly faster in spinal anesthesia group than in CSEA group, there was no significant difference in Apgar score between the groups. Moreover, failure of anesthesia and post-dural puncture headache were higher in spinal group than in CSEA group. CSEA is useful as anesthesia for an emergency cesarean section.


A large survey of Croatian wild mammals for Giardia duodenalis reveals a low prevalence and limited zoonotic potential.
Wild mammals are considered an important source of potentially zoonotic Giardia duodenalis parasites, yet surprisingly little information is available on the actual prevalence and the genetic identity of the species they harbor. A large survey was conducted in Croatia by collecting 832 fecal samples from red deer (Cervus elaphus, n = 374), roe deer (Capreolus capreolus, n = 21), wild boars (Sus scrofa, n = 144), foxes (Vulpes vulpes, n = 66), bears (Ursus arctos, n = 19), wolves (Canis lupus, n = 127), jackals (Canis aureus, n = 8), and hares (Lepus europeus, n = 73). Fecal samples were tested for the presence of Giardia cysts using fluorescent microscopy. The observed prevalence ranged from low (1% in red deer, 1.7% in wild boars, and 4.5% in foxes) to moderate (10% in wolves and 12.5% in jackals) to high (24% in roe deer). No cysts were observed in bears and hares. Polymerase chain reaction was performed on microscopically positive samples to amplify fragments of the small subunit ribosomal gene, the ribosomal 5.8S gene and the two flanking internal transcribed sequences, and the triose phosphate isomerase gene. Sequence analysis showed a predominance of G. duodenalis assemblage A in both ruminants (genotypes A1 and A3) and carnivores (genotype A1). G. duodenalis assemblages B, C, and D, as well as Giardia microti, were also detected in this study. This is the first molecular description of the parasite from the red deer, the wolf, and the jackal. The data point to a minor role of wild mammals as reservoirs of zoonotic assemblages of G. duodenalis, albeit cycling between sylvatic and domestic animals is possible.


A strand of cardiac muscle. Its ultrastructure and the electrophysiological implications of its geometry.
The structure of a small strand of rabbit heart muscle fibers (trabecula carnea), 30-80 micro in diameter, has been examined with light and electron microscopy. By establishing a correlation between the appearance of regions of close fiber contact in light and electron microscopy, the extent and distribution of regions of close apposition of fibers has been evaluated in approximately 200 micro length of a strand. The distribution of possible regions of resistive coupling between fibers has been approximated by a model system of cables. The theoretical linear electrical properties of such a system have been analyzed and the implications of the results of this analysis are discussed. Since this preparation is to be used for correlated studies of the electrical, mechanical, and cytochemical properties of cardiac muscle, a comprehensive study of the morphology of this preparation has been made. The muscle fibers in it are distinguished from those of the rabbit papillary muscle, in that they have no triads and have a kind of mitochondrion not found in papillary muscle. No evidence of a transverse tubular system was found, but junctions of cisternae of the sarcoplasmic reticulum and the sarcolemma, peripheral couplings, were present. The electrophysiological implications of the absence of transverse tubules are discussed. The cisternae of the couplings showed periodic tubular extensions toward the sarcolemma. A regularly spaced array of Z line-like material was observed, suggesting a possible mechanism for sarcomere growth.


Rapid Induction of Female-to-Male Sex Change in Adult Zebrafish by Injection of an Aromatase Inhibitor.
Previously, we examined whether aromatase inhibitor (AI) treatment induces a sex change in adult female zebrafish. A 5-month AI treatment regime resulted in the retraction of the ovaries and testis formation. Eight weeks after changing the diet to AI-free food, a large number of normal sperm were obtained. Artificial fertilization using sperm from the sex-changed females was successful. These results demonstrated that sex plasticity remains in the mature ovaries of zebrafish. However, >7 months of treatment was necessary; thus, pairing was unsuccessful. In this study, we tried to induce sex change through the injection of an AI to shorten the time course of sex change. When the AI solution was directly injected into the abdomen of zebrafish, retraction of the ovary was induced within 2 months. The natural mating of sex-changed females with normal females was successful at 3 months. Although the fertilization rate was low, juveniles resulting from these matings developed normally. We succeeded in establishing a method for inducing sex changes in adult zebrafish within 3 months. The procedure will support the study of how sexual plasticity persists in adult zebrafish following sex differentiation and the identification of undifferentiated stem cells.


Liver stiffness-based score in hepatoma risk assessment for chronic hepatitis C patients after successful antiviral therapy.
Patients with chronic hepatitis C (CHC) after successful antiviral therapy remain at risk of hepatocellular carcinoma (HCC). This study was to determine whether liver stiffness measurement (LSM) was useful in HCC risk assessment and to develop a risk-score system for clinical use. This retrospective study enrolled patients with CHC achieving sustained virological response (SVR) after interferon-based therapy with LSM at/after SVR determination. The demographics, clinical characteristics and HCC development were obtained from medical chart reviews. The diagnosis of HCC was based on recommended criteria. A total of 376 (M/F: 185/191, mean age: 54.1 years) patients, including 278 with pretreatment liver biopsy specimens, with a median follow-up period of 7.6 years were enrolled. Twenty-one patients developed HCC. The 5- and 10-year cumulative HCC incidences were 1.4% and 7.8%, respectively. Multivariate analysis showed advanced fibrosis/cirrhosis, diabetes and LSM were associated with HCC developments with odds ratio (OR) of 12.38, 2.80 and 1.01, respectively. For LSM in HCC prediction, the performance and cut-off were 0.783 and 12 kilopascal (kPa), respectively. For 278 patients with pretreatment biopsy, a risk-score system (score 0-4) combining advanced fibrosis/cirrhosis, diabetes and LSM >12 kPa was developed. With the low-risk group as a reference, patients in intermediate- (OR: 12.57) and high-risk (OR: 197.33) groups carried higher risk of HCC development. For patients with CHC achieving SVR, liver stiffness value at/after SVR determination was associated with HCC development independently. Patients with pretreatment advanced fibrosis/cirrhosis, diabetes and LSM >12 kPa after SVR were at high risk of HCC development.


[Mycophenolate mofetil in lupus nephritis].
The treatment of severe lupus nephritis is based on the combination of steroids and cytotoxic drugs. Intravenous cyclophosphamide administered in "pulses" is effective in the induction of remission but other therapeutic alternatives are sought in refractory cases or severely relapsing patients. Mycophenolate mofetil, used in renal transplantation, also can be useful in severe lupus nephritis. We describe the evolution of 6 patients (5 women and 1 man; age 17-45 years) with severe lupus nephropathy who after achieving remission with intravenous cyclophosphamide and steroids (5 cases) or cyclosporin A (1 case) showed relapse of proteinuria and were treated with mycophenolate mofetil (dose 1000-2000 mg/day). Two patients have completed 24 months, 1 patient two cycles of 12 months, 2 patients 18 months and 1 patient 6 months. After this treatment, all patients have achieved remission (3 partial and 3 complete). There was no treatment failure and no one patient discontinued medication; however 1 case relapsed. There were no changes in leucocytes, haemoglobin, serum creatinine and serum albumin. ANA and alpha DNA antibodies decreased. Proteinuria (measured as protein/creatinine urine ratio: initial 3 and final 0.3) and dose of steroids (initial: 17.5 mg/d and final 5 mg/d) decreased significantly (p < 0.05 Wilcoxon t-test). The most common side effects were nausea and abdominal discomfort that improved without discontinuation of treatment. We conclude that mycophenolate mofetil is effective and a safe drug in severe relapsing lupus nephritis.


Tracking of Diversity and Evolution in the Brown Rot Fungi Monilinia fructicola, Monilinia fructigena, and Monilinia laxa.
Monilinia species are among the most devastating fungi worldwide as they cause brown rot and blossom blight on fruit trees. To understand the molecular bases of their pathogenic lifestyles, we compared the newly assembled genomes of single strains of Monilinia fructicola, M. fructigena and M. laxa, with those of Botrytis cinerea and Sclerotinia sclerotiorum, as the closest species within Sclerotiniaceae. Phylogenomic analysis of orthologous proteins and syntenic investigation suggest that M. laxa is closer to M. fructigena than M. fructicola, and is closest to the other investigated Sclerotiniaceae species. This indicates that M. laxa was the earliest result of the speciation process. Distinct evolutionary profiles were observed for transposable elements (TEs). M. fructicola and M. laxa showed older bursts of TE insertions, which were affected (mainly in M. fructicola) by repeat-induced point (RIP) mutation gene silencing mechanisms. These suggested frequent occurrence of the sexual process in M. fructicola. More recent TE expansion linked with low RIP action was observed in M. fructigena, with very little in S. sclerotiorum and B. cinerea. The detection of active non-syntenic TEs is indicative of horizontal gene transfer and has resulted in alterations in specific gene functions. Analysis of candidate effectors, biosynthetic gene clusters for secondary metabolites and carbohydrate-active enzymes, indicated that Monilinia genus has multiple virulence mechanisms to infect host plants, including toxins, cell-death elicitor, putative virulence factors and cell-wall-degrading enzymes. Some species-specific pathogenic factors might explain differences in terms of host plant and organ preferences between M. fructigena and the other two Monilinia species.


Patient, carer and health service outcomes of nurse-led early discharge after breast cancer surgery: a randomised controlled trial.
Patients with breast cancer who require axillary clearance traditionally remain in hospital until their wound drains are removed. Early discharge has been shown to improve clinical outcomes, but there has been little assessment of the psychosocial and financial impact of early discharge on patients, carers and the health service. This study aimed to evaluate the effectiveness of a nurse-led model of early discharge from hospital. Main outcome measures were quality of life and carer burden. Secondary outcomes included patient satisfaction, arm morbidity, impact on community nurses, health service costs, surgical cancellations and in-patient nursing dependency. A total of 108 patients undergoing axillary clearance with mastectomy or wide local excision for breast cancer were randomised to nurse-led early discharge or conventional stay. Nurse-led early discharge had no adverse effects on quality of life or patient satisfaction, had little effect on carer burden, improved communication between primary and secondary care, reduced cancellations and was safely implemented in a mixed rural/urban setting. In total, 40% of eligible patients agreed to take part. Nonparticipants were significantly older, more likely to live alone and had lower emotional well being before surgery. This study provides further evidence of the benefits of early discharge from hospital following axillary clearance for breast cancer. However, if given the choice, most patients prefer to stay in hospital until their wound drains are removed.


Serological study of Japanese encephalitis in Nepal.
Antibodies against Japanese encephalitis virus (JEV) were examined in 356 serum samples from individuals visiting Tribhuvan University Teaching Hospital using neutralization test. A total of 13.2% (47/356) seropositivity was observed. Antibody positivity against both Nakayama-NIH and Beijing-1 strains was seen in 7.9% subjects. Infection by Nakayama-NIH strain alone was seen to be 4.2% while 1.1% of study populations were found to be infected only by Beijing-1 strain. High antibody titer was observed in the 20-40 years age-group and did not increase with age. Seropositivity was more common in zone-A (Tarai) area compared to zone-B (Hills) and zone-C (Mountains). Spectrum of antigenicity of JEV in zone-A as shown by neutralization test was identical to Indian isolates (strains). Difference in seropositivity was observed against Nakayama-NIH and Beijing-1 strain in zone-B and C indicating recent spread of JEV in these areas.


Differential Site-Based Expression of Pentose Phosphate Pathway-Related Proteins among Breast Cancer Metastases.
Purpose. We aimed to investigate the expression of pentose phosphate pathway- (PPP-) related proteins in metastatic breast cancer and its relationship with clinicopathologic factors. Methods. Tissue samples from 126 metastatic breast cancers were included in a tissue microarray. Expression of PPP-related proteins [glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconolactonase (6PGL), 6-phosphogluconate dehydrogenase (6PGDH), and nuclear factor erythroid 2-related factor (NRF2)] was determined by immunohistochemistry. Results. G6PDH (p = 0.011) and cytoplasmic NRF2 (p = 0.001) showed the highest expression in brain metastases. Human epidermal growth factor receptor (HER-2) positivity was associated with G6PDH (p < 0.001) and cytoplasmic NRF2 (p = 0.015) positivity. A high Ki-67 labeling index (LI) was correlated with nuclear NRF2 positivity (p = 0.037), and HER-2-positive luminal B type was associated with G6PDH positivity (p = 0.001). On multivariate Cox analysis, independent risk factors of short overall survival were 6PGL positivity in bone metastasis (HR 4.180, 95% CI 1.160-15.06, p = 0.029) and low Ki-67 LI in lung metastasis (HR 11.853, 95% CI 1.841-76.30, p = 0.009). Conclusion. Differential expression of PPP-related proteins correlated with different prognoses and metastatic sites, with the highest expression in brain metastases, and could be a potential therapeutic target.


Pulmonary and Regional Deposition of Nebulized and Dry Powder Aerosols in Ferrets.
The utilization of ferrets as a non-clinical model for disease is rapidly increasing within drug development. Many of these models include respiratory diseases that involve targeted drug delivery via nose-only inhalation. While the deposition patterns within other non-clinical models (mice, rats, canines, and non-human primates) have been well studied, the local and regional deposition of aerosols in ferrets has not been well characterized. Therefore, inhalation aerosols were developed, radiolabeled and the radiolabeling methods validated to support SPECT-CT imaging and quantification of regional deposition within ferrets. The studies were conducted with one liquid formulation and one dry powder formulation (two concentrations of dry powder). Additionally, both aerosols were polydisperse and therefore reflect the majority of pharmaceutical aerosols. Overall, the studies showed lung deposition fractions between 5 and 10% with median aerodynamic particle sizes of 2.5 and 2.8 μm. The lung deposition fraction of the liquid aerosol was ~ 9%, nearly double observed in rats with a similarly sized aerosol. Analysis of respiratory tract (oropharynx, laryngopharynx, trachea, bifurcation area, and lung) deposition indicates increased deposition of the liquid aerosol compared to the dry powder aerosol, however, when this analysis was refined to the pulmonary region (trachea, bifurcation, and lung) the deposition was similar between formulations. These data provide the first description of the regional deposition of inhalation aerosols in ferrets with standard nose-only inhalation procedures. These data can be used for calculations of both total and regional doses within ferret inhalation drug delivery.


Molecular self-assembly at metal-electrolyte interfaces.
The self-assembly of molecular layers has become an important strategy in modern design of functional materials. However, in particular, large organic molecules may no longer be sufficiently volatile to be deposited by vapor deposition. In this case, deposition from solution may be a promising route; in ionic form, these molecules may even be soluble in water. In this contribution, we present and discuss results on the electrochemical deposition of viologen- and porphyrin molecules as well as their co-adsorption on chloride modified Cu(100) and Cu(111) single crystal electrode surfaces from aqueous acidic solutions. Using in situ techniques like cyclic voltametry and high resolution scanning tunneling microscopy, as well as ex-situ photoelectron spectroscopy data the highly ordered self-assembled organic layers are characterized with respect to their electrochemical behavior, lateral order and inner conformation as well as phase transitions thereof as a function of their redox-state and the symmetry of the substrate. As a result, detailed structure models are derived and are discussed in terms of the prevailing interactions.


Analysis of the Candida albicans - specific T-cell response and oropharyngeal Candida colonization in a cohort of HIV-1-infected patients.
To investigate Candida epidemiology and immunologic correlates of protection in HIV-1 infected patients, we analyzed oral Candida colonization in correlation to the Candida-specific T-cell response measured by g-IFN ELISPOT using different Candida (C.) albicans strains. In 16/46 patients (13 asymptomatic, 3 with oral thrush), but in 0/28 controls, Candida (13 C. albicans, 1 C. lusitaniae, 1 C. krusei, 1 C. parapsilosis) was isolated. Candida specific T-cells were detected more frequently in controls (20/28) than in HIV-1+ subjects (16/46, p= 0.03). We observed a significant association of higher CD4 cell numbers with both detection of Candida specific T-cells and lack of oral Candida colonization, but there was no significant correlation of oral Candida colonization to the detection of Candida specific T-cells, viral load or antiretroviral therapy. Thus, local mucosal immunity seems to be more important in the pathogenesis of Candida colonization than circulating Candida specific T-cells. The pathogenic C. albicans strain K24122 was less frequently recognized by patients (6/46) than the laboratory adapted strain SC5314 (14/46, p= 0.03), whereas a similar recognition of both strains was observed in healthy controls. This indicates an impaired Candida-specific T-cell repertoire in HIV+ patients that could increase the risk of immune evasion by C. albicans.


Importance of post-translational modifications on the function of key haemostatic proteins.
Post-translational modifications (PTMs) such as glycosylation and phosphorylation play an important role on the function of haemostatic proteins and are critical in the setting of disease. Such secondary level changes to haemostatic proteins have wide ranging effects on their ability to interact with other proteins. This review aimed to summarize the knowledge of the common PTMs associated with haemostatic proteins and the implications of such modifications on protein function. Haemostatic proteins that represent the main focus for studies specific to PTMs are von Willebrand factor, tissue factor, factor VIII, antithrombin and fibrinogen. These proteins are susceptible to PTMs by glycosylation, phosphorylation, sulphation, citrullination and nitration, respectively, with a significant impact on their function. During synthesis, vWF must undergo extensive PTMs, with N-linked glycosylation being the most common. Increased phosphorylation of tissue factor results in increased affinity for platelets to the vessel endothelium. Citrullination of antithrombin leads to an increased anticoagulant function of this protein and therefore an anticoagulant state that inhibits clot formation. On the contrary, nitration of fibrinogen has been shown to result in a prothrombotic state, whilst sulphation is required for the normal function of Factor VIII. From this review, it is evident that PTMs of haemostatic proteins as a change in protein structure at a secondary level greatly influences the behaviour of the protein at a tertiary level.


Making and breaking a health service.
The creation of the National Health Service (NHS) marked a radical break with the past, making health care universally available on the basis of need rather than means. The NHS was conceived during wartime emergency and has had to survive further regular crises to reach its 70th year, but it now faces challenges that are unprecedented in scale and there are doubts about its ability to continue in its present form. Resources have not increased with need, and the NHS can no longer function as a comprehensive service during periods of peak demand. Policymakers look for solutions in service rearrangements, new technologies, quality improvement initiatives and alternative funding arrangements; meanwhile, chronic lack of capacity is taking a predictable toll on patient care and staff morale. The NHS has become a formidably resilient institution, but securing its future may take as great a collective effort as the one that created it.


[Current situation and cause analysis of physical activity in Chinese primary and middle school students in 2010].
To analyze the characteristics of physical activity in Chinese primary and middle school students, and to provide scientific basis for school physical education and health work. The data were collected from 166 812 Han primary school pupils above grade 4, junior and senior middle school students who participated in 2010 National Physical Fitness and Health Surveillance in 30 provinces (excluding autonomous regions, municipalities). Nationwide in 2010, 22.7% of the students had been physically active doing any kind of physical activity for a total of at least 60 minutes per day, the prevalence rates of the 9 to 13, 13 to 16, 16 to 19 age groups were 32.7%, 20.7% and 12.5%, respectively (P<0.01). The prevalence of having been physically active for a total of at least 60 minutes per day was higher among the Eastern Region's students than the Western Region's students in the different age groups (P<0.05), of whom 61.8% reported that they sweated and felt a little bit tired in physical education, 68.6% thought that they liked their physical education, 72.4% were willing to take part in extracurricular physical activities organized by their school, and 46.4% were willing to participate in the long distance running exercise. "Fear of fatigue or/and hardship" was the most important reason why the students did not actively participate in physical activities and long-distance running exercise among both the boys and girls in the different age groups. The prevalence of having been physically active for a total of at least 60 minutes per day in Chinese students is still low, and there is an obvious difference in the different groups. In the future, not only should the students' physical exercise time and intensity be further guaranteed, but also their awareness strengthened and their habits of physical exercise cultured.


Hybrid modeling and prediction of dynamical systems.
Scientific analysis often relies on the ability to make accurate predictions of a system's dynamics. Mechanistic models, parameterized by a number of unknown parameters, are often used for this purpose. Accurate estimation of the model state and parameters prior to prediction is necessary, but may be complicated by issues such as noisy data and uncertainty in parameters and initial conditions. At the other end of the spectrum exist nonparametric methods, which rely solely on data to build their predictions. While these nonparametric methods do not require a model of the system, their performance is strongly influenced by the amount and noisiness of the data. In this article, we consider a hybrid approach to modeling and prediction which merges recent advancements in nonparametric analysis with standard parametric methods. The general idea is to replace a subset of a mechanistic model's equations with their corresponding nonparametric representations, resulting in a hybrid modeling and prediction scheme. Overall, we find that this hybrid approach allows for more robust parameter estimation and improved short-term prediction in situations where there is a large uncertainty in model parameters. We demonstrate these advantages in the classical Lorenz-63 chaotic system and in networks of Hindmarsh-Rose neurons before application to experimentally collected structured population data.


A survey of renal impairment pharmacokinetic studies for new oncology drug approvals in the USA from 2010 to early 2015: a focus on development strategies and future directions.
The US Food and Drug Administration (FDA) issued a guidance document in 2010 on pharmacokinetic (PK) studies in renal impairment (RI) on the basis of observations that substances such as uremic toxins might result in altered drug metabolism and excretion. No specific recommendations for oncology drugs were included. We surveyed the publicly available FDA review documents of 29 small molecule oncology drugs approved between 2010 and the first quarter of 2015. The objectives were as follows: (i) summarize the impact of RI on PK at the time of the initial new drug application; (ii) identify limitations of the guidance; and (iii) outline an integrated approach to study the impact of RI on these drugs. Our survey indicates that the current FDA guidance does not appear to provide clear strategic or decision pathways for RI studies in terms of small molecule oncology drugs. The FDA review documents indicate an individualized approach to the review because of the complex pharmacologic nature of these drugs and patient populations. Overall, the strategy for carrying out a RI study during clinical development or as a postmarketing study requires integration with the totality of data, including mass balance, absolute bioavailability, drug-drug interaction, hepatic dysfunction, population PK, exposure-response analysis, the therapeutic window for best guidance, and determination of the optimal doses for special oncology populations.


[Early diagnosis of periodontal disease].
Early diagnosis is necessary if the prevalence of periodontal disease is to be brought under control. For periodontal screening in dental practice we propose to employ the C.P.I.T.N. system, combined with examination of two or four dental bite-wing radiographs. Both of these procedures can be accurately applied in general dental practice conditions. The screening is not time consuming and can yield very important diagnostic data.


Gamma-interferon exerts a critical early restriction on replication and dissemination of yellow fever virus vaccine strain 17D-204.
Live attenuated viruses are historically among the most effective viral vaccines. Development of a safe vaccine requires the virus to be less virulent, a phenotype that is historically arrived by empirical evaluation often leaving the mechanisms of attenuation unknown. The yellow fever virus 17D live attenuated vaccine strain has been developed as a delivery vector for heterologous antigens; however, the mechanisms of attenuation remain elusive. The successful and safe progress of 17D as a vaccine vector and the development of live attenuated vaccines (LAVs) to related flaviviruses requires an understanding of the molecular mechanisms leading to attenuation. Using subcutaneous infection of interferon-deficient mouse models of wild type yellow fever virus (WT YFV) pathogenesis and 17D-mediated immunity, we found that, in the absence of type I IFN (IFN-α/β), type II interferon (IFN-γ) restricted 17D replication, but not that of WT YFV, by 1-2 days post-infection. In this context, IFN-γ responses protected 17D-infected animals from mortality, largely restricted the virus to lymphoid organs, and eliminated viscerotropic disease signs such as steatosis in the liver and inflammatory cell infiltration into the spleen. However, WT YFV caused a disseminated infection, gross liver pathology, and rapid death of the animals. In vitro, IFN-γ treatment of myeloid cells suppressed the replication of 17D significantly more than that of WT YFV, suggesting a direct differential effect on 17D virus replication. Together these data indicate that an important mechanism of 17D attenuation in vivo is increased sensitivity to IFN-γ stimulated responses elicited early after infection.


Immunity, autoimmunity and autoimmune diseases in older people.
Immunosenescence is defined as all the changes occurring in the immune system in the aged. Recent studies suggest that in older patients the immune system undergoes a functional remodelling. Two contrasting phenomena coexist in immunosenescence: the decreasing of immune response and the autoantibody production. Possible consequences are an increase of autoimmune phenomenon, neoplasia incidence, and predisposition to infections. The study of autoimmune manifestations in the elderly population should be considered as a priority for future medical research because of the increase in life expectancy, especially in developed countries. This review analyzes the clinical expression of systemic autoimmune diseases in older patients.


An expression vector for the archaebacterium Haloferax volcanii.
The recent development of an efficient transformation method and shuttle vectors for Haloferax volcanii has set the stage for rapid progress in archaebacterial molecular biology. We describe a shuttle-expression vector that can be selected for and maintained in either H. volcanii or Escherichia coli and permits the expression of cloned genes in H. volcanii. The vector, pWL204, was constructed by incorporating an H. volcanii tRNA(Lys) gene promoter into a derivative of the H. volcanii-E. coli shuttle vector pWL102. The vector has been used to express a modified, intron-containing, H. mediterranei tRNA(Trp) gene (tRNA(Trp)-O167). Transcription from the tRNA(Lys) gene promoter in vivo was detected by Northern (RNA) analysis with an oligonucleotide probe complementary to the unique intron sequence of tRNA(Trp)-O167. Dependence of transcription on the tRNA(Lys) promoter was demonstrated by the absence of transcription when the promoter sequence was deleted from the vector and by mapping the transcription initiation site by primer extension.


Relationship between protein kinase C alterations and nitric oxide overproduction in cirrhotic rat aortas.
Although nitric oxide (NO) overproduction and protein kinase C (PKC) alterations may play a role in systemic haemodynamic changes in cirrhotic rat aortas, the relationship between NO synthase (NOS) hyperactivation and PKC hypoactivation is unknown. Therefore, the relationships between NOS and PKC activities were studied in cirrhotic rat aortas. The effects of NOS inhibition by Nw-nitro-L-arginine (LNNA) on the contractile response to phorbol myristate acetate (PMA), a PKC activator, were studied. The effects of NOS inhibition and those of S-nitroso-N acetyl-DL-penicillamine (SNAP), an NO donor, on PKC activity were also evaluated. The effects of PKC activation and inhibition on total NOS and inducible NOS (iNOS) activities were measured. Nitric oxide synthase inhibition caused an increase in PMA-induced contraction and an increase in PKC activity in cirrhotic rat aortas. S-nitroso-N acetyl-DL-penicillamine induced downregulation of PKC activity. Total basal aortic NOS activity was significantly higher in cirrhotic rats than in control rats and activation of PKC by PMA induced a decrease in total aortic NOS activity. Protein kinase C downregulation caused an increase in both total aortic NOS and iNOS activities only in control rats, whereas only iNOS activity increased in cirrhotic rats. In cirrhotic rat aortas, NO overproduction plays a role in the decreased PKC activation that leads to reduced aortic contraction. Overactivation of aortic NOS in cirrhotic rats may be because of, in part, the reduced PKC activity.


The phenylic hydroxyl group is essential for the induction of stress response by sodium salicylate.
We have shown that sodium salicylate (SA) activates the heat shock promoter and induces the expression of heat shock proteins (Hsps) with a concomitant increase in the thermotolerance of cells. To identify the functional groups of SA necessary for the induction of Hsps, we evaluated the effect of various derivatives of SA using a mammalian cell line containing a reporter gene downstream of an hsp105 promoter. Among the derivatives, the compounds in which the carboxyl group of SA was substituted activated the hsp105 promoter at 37 degrees C as SA did, but the compounds in which the hydroxyl group was substituted did not. Thus, the phenylic hydroxyl group but not the carboxyl group of SA seemed to be necessary for a stress-induced response. In addition, the orientation of two functional groups on the benzene ring of SA derivatives was also important for the induction of a response. Among these compounds, salicylalcohol which strongly induced the expression of Hsps suppressed the protein aggregation and apoptosis caused by an expanded polyglutamine tract in a cellular model of polyglutamine disease. These findings may aid in the development of novel effective Hsp-inducers.


Allogeneic hematopoietic cell transplantation for myelofibrosis in the era of JAK inhibitors.
The discovery of JAK2617F mutation paved the way for the development of small molecule inhibitors of JAK1/2 resulting in first approved JAK1/2 inhibitor, ruxolitinib, for the treatment of patients with myelofibrosis (MF). Although JAK1/2 inhibitor therapy is effective in decreasing the burden of symptoms associated with splenomegaly and MF-related constitutional symptoms, it is neither curative nor effective in reducing the risk of leukemic transformation. Presently, allogeneic hematopoietic cell transplantation (HCT) is the only curative therapy for MF. A significant risk of regimen-related toxicities, graft failure, and GVHD are major barriers to the success of HCT in MF. Because of significant HCT-associated morbidity and mortality, divergent opinions regarding its appropriate role in this clinical situation have emerged. In this review, the risk-benefit ratios of modern drug therapy compared with HCT in MF patients are analyzed. A risk-adapted approach individualized to each patient's biologic characteristics and comorbidities is described, which is currently warranted in determining optimal treatment strategies for patients with MF. Inclusion of JAK1/2 inhibitor therapy in future transplant conditioning regimens may provide an opportunity to overcome some of these barriers, resulting in greater success with HCT for MF patients.


Multiple intravesical instillations of low-dose resiniferatoxin in the treatment of refractory interstitial cystitis.
Current medications used in the treatment of interstitial cystitis (IC) have limited efficacy. This prospective study investigated the efficacy of multiple intravesical instillations of resiniferatoxin (RTX) at the concentration of 10 nM. Patients with proven IC previously treated with traditional medications for more than 6 months without clinical benefit were enrolled. They were excluded if bladder outlet obstruction or urinary tract infection was present. Intravesical instillation of low-dose RTX (10 nM) once weekly for 4 weeks was performed at the outpatient department. International Prostate Symptom Score (IPSS), 5-Point Pain Scale, and Quality of Life Index (QOL Index) were recorded. A videourodynamic study was done at baseline and 3 months after treatment. The therapeutic results and urodynamic parameters were compared between baseline and 3 months. Thirteen patients, including 10 women and 3 men, were enrolled in this study. The mean duration of IC symptoms was 4.4 +/- 2.5 years. The mean duration of active treatment was 15.3 +/- 8.6 months. One female patient dropped out due to severe bladder pain after RTX instillation. Among the 12 patients who completed the study treatment, subjective assessment revealed that 2 had an excellent therapeutic result, 5 had an improved result and 5 remained unchanged from baseline. The overall satisfactory rate was 58.3%. IPSS, 5-Point Pain Scale, and QOL Index were significantly decreased after RTX treatment. There was no significant increase in mean functional bladder capacity or change in urodynamic parameters. No serious adverse event occurred after RTX treatment. This study suggests that multiple intravesical instillations of RTX at the concentration of 10 nM are effective in relieving lower urinary tract symptoms in patients with refractory IC. The treatment is tolerable and suitable for use on an outpatient clinic basis.


Second marrow transplants in patients with leukemia who relapse after allogeneic marrow transplantation.
Twenty-six patients with recurrent leukemia following allogeneic marrow transplantation received a second marrow transplant between 1.5 and 78 months (median 26) after the initial transplant. Preparative regimens for second transplant included multi-agent chemotherapy with total body irradiation, 2.0-10.0 Gy (five patients), dimethylbusulfan alone (one patient), and dimethylbusulfan or busulfan plus cyclophosphamide (20 patients). One patient died before engraftment of infection and 18 died after engraftment from veno-occlusive disease (4), infection (2), idiopathic pneumonia (3), cytomegalovirus pneumonia (3), leukemia (5) and encephalopathy (1). Seven patients (27%) survive 12-38 months (median 26); five (19%) are disease-free and two have recurrent leukemia. Two of the five disease-free survivors have chronic graft-versus-host disease. All of the surviving patients received dimethylbusulfan or busulfan plus cyclophosphamide and six of the seven surviving patients were among 11 patients transplanted more than 2 years after the first transplant whereas only one was among the 15 transplanted in less than 2 years. Those who have second marrow transplants one or more years after their initial transplant are more likely to benefit, while those who are less than 1 year from initial transplant appear to benefit the least.


A comparison of oral, axillary, rectal and tympanic-membrane temperatures of intensive care patients with and without an oral endotracheal tube.
Monitoring rectal temperatures (RTs) in orally intubated, critically ill patients is common practice, as they are generally believed to be more accurate than oral temperatures (OTs). This study was designed to compare OTs of critically ill patients, with and without an endotracheal tube, using patients as their own control. Axillary (AT), tympanic membrane (TMT) and RTs of patients were also studied. Sixty-five orally intubated patients ready for extubation, ranging in age from 17 to 96 years (17 female and 48 male), were included in the study. OT, AT, RT and TMTs were taken before and after extubation. Using a t-test for paired samples, significant differences were found between both ATs and OTs measured with and without an oral endotracheal tube (P < 0.01); the differences (-0.12 and 0.08 degrees C, respectively) were not considered clinically important. Further t-tests for paired samples revealed no significant difference between TMTs and RTs pre and post extubation. Respiratory rate, presence or absence of teeth, and mouth position were shown to have no effect on OT (t-tests, P < 0.01). Analysis of variance showed no difference in OT dependent on room temperature or temperature within the endotracheal tube. It was concluded that OTs are accurate when taken in patients who are orally intubated and ready for extubation.


Priming with copper-chitosan nanoparticles elicit tolerance against PEG-induced hyperosmotic stress and salinity in wheat.
In this study Cu-chitosan nanoparticles (Cu-CNP) have been employed as eco-friendly and safer priming agents to induce salt and PEG-induced hyperosmotic stress tolerance in wheat seedlings. Seed priming is a facile on-farm stress management technique that requires a little amount of priming agent and minimizes the eco-toxicological effects on soil fertility. The wheat seeds were primed with 0.12% and 0.16% Cu-CNP for eight hours and were allowed to germinate under normal, PEG-induced hyperosmotic stress (15% PEG-6000 - 3.0 Mpa) and salt stress (150 mM). For comparison, non-primed and hydro-primed seeds were also allowed to germinate as control under the same conditions. The biochemical analyses suggested the priming treatments enhanced the POD activity under salt stress but it was decreased under PEG-induced hyperosmotic stress. Priming with 0.12% Cu-CNP induced a significant increase in CAT while the opposite effect was observed in 0.16% treated seedling under stress and non-stress conditions. Both priming treatments did not allow the over-expression of SOD under both stress conditions. The total phenolic contents were also decreased significantly under all conditions. Except for priming with 0.16% Cu-CNP under PEG-induced hyperosmotic stress, a suppression in MDA was observed under both stress conditions. Surprisingly, the Cu-CNP priming induced a significant increase in β-carotenoids, total carotenoids, chlorophyll a, b and total chlorophyll under normal and stress conditions. In conclusion, the controlled expression of enzymatic antioxidants, low contents of non-enzymatic antioxidants and suppression of MDA mirror the stress mitigating role of Cu-CNP against PEG-induced hyperosmotic stress and salinity. The stress-insulating potential has also been reinforced by the enhanced production of plant and photosynthetic pigments. All these priming-induced biochemical changes produced positive effects on growth and germinating parameters in wheat seedlings under PEG-induced hyperosmotic stress as well as salinity.


Systematic computerised cardiovascular health screening for people with severe mental illness.
Aims and method People with severe mental illness (SMI) die relatively young, with mortality rates four times higher than average, mainly from natural causes, including heart disease. We developed a computer-based physical health screening template for use with primary care information systems and evaluated its introduction across a whole city against standards recommended by the National Institute for Health and Care Excellence for physical health and cardiovascular risk screening. Results A significant proportion of SMI patients were excluded from the SMI register and only a third of people on the register had an annual physical health check recorded. The screening template was taken up by 75% of GP practices and was associated with better quality screening than usual care, doubling the rate of cardiovascular risk recording and the early detection of high cardiovascular risk. Clinical implications A computerised annual physical health screening template can be introduced to clinical information systems to improve quality of care.


Identifying early modern human ecological niche expansions and associated cultural dynamics in the South African Middle Stone Age.
The archaeological record shows that typically human cultural traits emerged at different times, in different parts of the world, and among different hominin taxa. This pattern suggests that their emergence is the outcome of complex and nonlinear evolutionary trajectories, influenced by environmental, demographic, and social factors, that need to be understood and traced at regional scales. The application of predictive algorithms using archaeological and paleoenvironmental data allows one to estimate the ecological niches occupied by past human populations and identify niche changes through time, thus providing the possibility of investigating relationships between cultural innovations and possible niche shifts. By using such methods to examine two key southern Africa archaeological cultures, the Still Bay [76-71 thousand years before present (ka)] and the Howiesons Poort (HP; 66-59 ka), we identify a niche shift characterized by a significant expansion in the breadth of the HP ecological niche. This expansion is coincident with aridification occurring across Marine Isotope Stage 4 (ca. 72-60 ka) and especially pronounced at 60 ka. We argue that this niche shift was made possible by the development of a flexible technological system, reliant on composite tools and cultural transmission strategies based more on "product copying" rather than "process copying." These results counter the one niche/one human taxon equation. They indicate that what makes our cultures, and probably the cultures of other members of our lineage, unique is their flexibility and ability to produce innovations that allow a population to shift its ecological niche.


Stepwise click functionalization of DNA through a bifunctional azide with a chelating and a nonchelating azido group.
A stepwise chemoselective click reaction was performed on nucleosides and oligonucleotides containing 7-octadiynyl-7-deaza-2'-deoxyguanosine and 5-octadiynyl-2'-deoxycytidine with unsymmetrical 2,5-bis(azidomethyl)pyridine using copper(II) acetate. The reaction is selective for the chelating azido group, thereby forming monofunctionalized adducts still carrying the nonchelating azido functionality. The azido-functionalized adduct was applied to a second click reaction, now performed in the presence of reducing agent, to generate cross-linked DNA or a pyrene click conjugate. The chelate-controlled stepwise click reaction is applicable to alkynylated nucleosides and oligonucleotides.


Liver transplantation for neuropsychiatric Wilson disease.
Although neuropsychiatric manifestations are prominent in some patients with Wilson disease, there is little published information regarding the efficacy of liver transplantation for these patients. A 22-year-old male with advanced neurological impairment and prominent psychiatric manifestations due to Wilson disease who underwent liver transplantation is presented. After transplantation, the ceruloplasmin and copper studies normalized and eventually the Kayser-Fleischer rings disappeared. Neurological recovery was very slow and incomplete, and his behavioural and personality disorder was entirely unaffected. He committed suicide 43 months post-transplantation. A review of the small number of related published cases in the English language literature shows variable neurological recovery post-transplantation, but the course of psychiatric manifestations is virtually never described. This case suggests that one must be cautious regarding liver transplantation for Wilson disease in patients with prior psychiatric manifestations. Aggressive medical management is likely to be preferable in most cases.


The Etiology of Improved Outcomes at High Volume Centers Learning Theory and the Case of Implant Flashing.
Increased volume has been shown to be associated with improved outcomes for many orthopaedic procedures. For individual surgeons, the concepts of learning curves and volume effects have been well established in the literature. For institutions, high-volume hospitals have also been shown to have better outcomes for orthopaedic procedures such as total joint replacements. However, exactly how hospital volume mediates this improvement is not well understood. Learning theory states that learning occurs as a result of accumulated experience, not based on time. We compared our institution's curve representing our implant flashing rates to other institutional data sets that exhibit learning and continuous quality improvement, including airline near misses, coal mining accidents, and others. Development of expertise is based on volume and rate of errors, and therefore higher volume is conducive to faster learning.


Prolonged in vivo functional assessment of the mouse oviduct using optical coherence tomography through a dorsal imaging window.
The oviduct (or fallopian tube) serves as an environment for gamete transport, fertilization and preimplantation embryo development in mammals. Although there has been increasing evidence linking infertility with disrupted oviduct function, the specific roles that the oviduct plays in both normal and impaired reproductive processes remain unclear. The mouse is an important mammalian model to study human reproduction. However, most of the current analyses of the mouse oviduct rely on static histology or 2D visualization, and are unable to provide dynamic and volumetric characterization of this organ. The lack of imaging access prevents longitudinal live analysis of the oviduct and its associated reproductive events, limiting the understanding of mechanistic aspects of fertilization and preimplantation pregnancy. To address this limitation, we report a 3D imaging approach that enables prolonged functional assessment of the mouse oviduct in vivo. By combining optical coherence tomography with a dorsal imaging window, this method allows for extended volumetric visualization of the oviduct dynamics, which was previously not achievable. The approach is used for quantitative analysis of oviduct contraction, spatiotemporal characterization of cilia beat frequency and longitudinal imaging. This new approach is a useful in vivo imaging platform for a variety of live studies in mammalian reproduction.


Net ecosystem carbon and greenhouse gas budgets in fiber and cereal cropping systems.
To assess the contributions of fiber and cereal production on climate change, the net ecosystem exchange of carbon dioxide (CO2), main exchanges of non-CO2 carbon, and methane (CH4) and nitrous oxide (N2O) fluxes were continuously monitored throughout two year-round crop cycles (Y1 and Y2: 1st and 2nd year-round crop cycles, respectively) using eddy covariance, biometric observation, and static chamber methods in typical cotton and wheat-maize rotational cropping systems in China. The evaluation of net ecosystem carbon budgets (NECBs: considering net ecosystem CO2 exchange and non-CO2 carbon exchanges by fertilization, seeding, and harvest) and greenhouse gas budgets (GHGBs: adding CH4 and N2O fluxes to the NECBs based on CO2 equivalents) showed that the cotton cropping system persistently functioned as an intensive carbon (-1527 and -974 kg C ha-1 yr-1) and greenhouse gas (GHG) source (5618 and 3591 kg CO2-eq ha-1 yr-1) because of the large CO2 emissions during the long fallow periods (5748 and 5160 kg CO2 ha-1 in Y1 and Y2, respectively). The wheat-maize cropping system had high net ecosystem production (NEP) and low harvest index and therefore, served as a notable carbon sink (1461 kg C ha-1 yr-1 in Y2). Although high irrigation water and chemical fertilizer inputs stimulated N2O emissions, the wheat-maize cropping system still behaved as an important GHG sink (-4257 kg CO2-eq ha-1 yr-1 in Y2) because of the tremendous net carbon sequestration. However, in Y1 incidental wind damage lowered the NEP and turned the wheat-maize cropping system into a GHG source (2144 kg CO2-eq ha-1 yr-1). The NEP, NECBs, and GHGBs of the double cropping system generally exceeded those of the single cropping system. The traditional rotation between double and single cropping systems should be restored to maintain soil carbon storage and alleviate the radiative forcing effects of cotton production.


Identification and characterization of genes encoding sex pheromone cAM373 activity in Enterococcus faecalis and Staphylococcus aureus.
The sex pheromone cAM373 of Enterococcus faecalis and the related staph-cAM373 of Staphylococcus aureus were found to correspond to heptapeptides located within the C-termini of the signal sequences of putative prelipoproteins. The deduced mature forms of the lipoproteins share no detectable homology and presumably serve unrelated functions in the cells. The chromosomally encoded genetic determinants for production of the pheromones have been identified and designated camE (encoding cAM373) and camS (encoding staph-cAM373). Truncated and full-length clones of camE were generated in Escherichia coli, in which cAM373 activity was expressed. In E. faecalis, insertional inactivation in the middle of camE had no detectable phenotypic effects on the pheromone system. Establishment of an in frame translation stop codon within the signal sequence resulted in reduction of cAM373 activity to 3% of normal levels. The camS determinant has homologues in Staphylococcus epidermidis, Bacillus subtilis and Listeria monocytogenes; however, corresponding heptapeptides present within those sequences do not resemble staph-cAM373 closely. The particular significance of staph-cAM373 as a potential intergeneric inducer of transfer-proficient genetic elements is discussed.


Neck circumference in adolescents and cardiometabolic risk: A sistematic review.
To critically analyze articles on the relation between neck circumference (NC) in adolescents and: body mass index, fat distribution, metabolic syndrome and its individual components, and cardiovascular risk. Systematic review undertaken by two independent researchers using the Pubmed/Medline, Lilacs/Medline, Scielo and Cochrane databases in English, Spanish and Portuguese in the period comprising the past 5 years. Eighteen (18) articles were selected. The articles show an association between NC in adolescents and body fat (BMI), central fat distribution (WC), metabolic syndrome and several of its individual components, and cardiovascular risk. Some values are proposed for NC cutoff points as a diagnostic tool for nutritional status, high blood pressure and pre-hypertension, cardiovascular risk, insulin resistance and metabolic syndrome. We identified a percentile curve constructed for Brazilian adolescents. There is a shortage of studies with representative samples, variety at the NC measurement sites, and the age of the participants, which makes it difficult to establish definitive landmarks.


Effect of SA4503, a novel sigma1 receptor agonist, against glutamate neurotoxicity in cultured rat retinal neurons.
We examined the effects of sigma1 receptor agonists against glutamate-induced neurotoxicity in cultured retinal neurons. Primary cultures obtained from fetal rat retinas (16-19 d gestation) were used. The neurotoxic effect of glutamate was quantitatively assessed using the trypan blue exclusion method. A brief exposure of retinal cultures to glutamate (500 microM) led to delayed neuronal cell death. The glutamate-induced neurotoxicity was inhibited by (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo-[a,b]-cyclohepten-5 ,10-imine hydrogen maleate (MK-801). The sigma1 receptor agonists, 1-(3,4-dimethoxyphenethyl)-4-(3-phenylpropyl)-piperazine dihydrochloride (SA4503) and (+)-pentazocine at a concentration range of 0.1 approximately 100 microM reduced the glutamate-induced neurotoxicity in a dose-dependent manner. In addition, the neuroprotective effects of both SA4503 and (+)-pentazocine were antagonized by co-treatment with N,N-dipropyl-2-[4-methoxy-3-(2-phenylethoxy)phenyl]ethylamine monohydrochloride (NE-100), a putative sigma1 receptor antagonist. These findings suggest that sigma1 receptor agonists protect retinal cells against glutamate-induced neurotoxicity.


External pneumatic intermittent compression for treatment of dependent pregnancy edema.
A portable external pneumatic intermittent compression (EPIC) device has been successful in reducing peripheral edema. This study explored the effectiveness of EPIC for treating dependent pregnancy edema. In the study, 42 healthy pregnant women received EPIC for 30 minutes at 40 torr while in the left lateral recumbent position: Group One with mid-thigh boots, and Group Two with below-knee boots. Prior to compression, descriptive data were gathered, leg circumference measurements made, and surface skin temperatures recorded for three sites per leg. Vital signs were taken and pedal edema subjectively indexed. Following compression, circumferences, skin temperatures, vital signs, and edema indices were rerecorded. Three volumes were calculated for each leg using a mathematical model of leg segments as conical frustum units. Mean volume reductions for each leg were significant. The mid-thigh-length boots produced greater mean volume decreases. The volume decrease for calf, lower leg, and foot frustum units were significant. EPIC holds promise as a useful treatment for dependent pregnancy edema.


Salience of alcohol expectancies and drinking outcomes.
There is a body of research which indicates that endorsement of positive alcohol expectancies predicts alcohol drinking patterns; however, there is inconsistency in the literature regarding which particular alcohol expectancies predict drinking. Although an individual may endorse a variety of alcohol expectancy beliefs, these beliefs may not be of equal importance to drinking decisions. This study investigated whether the prediction of drinking might be enhanced by considering salience of alcohol expectancies rather than mere endorsement. Hierarchical regression analyses demonstrated that expectancy salience significantly improved the prediction of total alcohol consumption above and beyond the effects of expectancy endorsement. Expectancy salience was less effective as a predictor of heavy drinking.


Correlation between Post-Thaw Spermatozoa Quality of the Endangered Javan Banteng with OPN Gene Expression.
The role of ex situ conservation facilities or captivity through captive breeding programs is essential in the conservation of the endangered Javan banteng. The development of semen cryopreservation may assist on one side of the conservation plan. However, the male Javan banteng reproductive capability must be considered as it influences the targeted outputs. Studying the potential biomarker for fertility such as osteopontin gene expression is also expected to help predict male fertility. Therefore, this study aimed to analyze the quality of spermatozoa after thawing to help predict the male reproductive capability of Javan banteng. Furthermore, this study investigated the potential role of osteopontin gene expression in male Javan banteng fertility. A positive reinforcement approach was used to accustom the male and female animals as we focused on establishing a collection procedure using neither sedation nor anaesthesia. Semen samples were collected at Taman Safari Indonesia, Bogor, in accordance with the female banteng receptivity. Semen samples were then evaluated and then cryopreserved under field conditions. Our study showed the different predicted reproductive capability of the Javan banteng based on the post-thaw spermatozoa quality, which showed significant differences. The OPN gene showed positive correlations with the progressive motility (r = 0.711, p = 0.048), viability (r = 0.822, p = 0.012), and acrosomal integrity (r = 0.665, p = 0.072) of Javan banteng spermatozoa after thawing. Our study demonstrated the predicted Javan banteng reproductive capability based on various post-thaw spermatozoa variables. This finding is also the first report on the OPN gene potential to be developed as the assessment tool of post-thaw spermatozoa quality of the male Javan banteng. The findings in our study may help give recommendations for future breeding programs, especially in the ex situ conservation sites.


Establishment of the BacMam system using silkworm baculovirus.
The BacMam system uses modified insect viruses (baculoviruses) as vehicles to efficiently deliver genes for expression in mammalian cells. The technique can be widely applied to large-scale recombinant protein production with appropriate modifications, high-throughput screening platforms for cell-based assays, and the delivery of large genes. The silkworm system is often employed as a rapid and cost-effective approach for recombinant baculovirus generation. Here we have developed the novel BacMam system using silkworm baculovirus, and shown the successful expression of EGFP in mammalian cells. The transduction to mammalian cells via the BacMam system was improved by adding phosphate-buffered saline and sodium butyrate to the culture medium and lowering the temperature after viral infection. This study provides an alternative gene delivery system for mammalian cells, which has various potential applications, including efficient native protein production and gene therapy.


CpG ODN-induced matrix metalloproteinase-13 expression is mediated via activation of the ERK and NF-κB signalling pathways in odontoblast cells.
To investigate the effects of CpG ODN (CpG oligodeoxynucleotides) to model the action of bacterial challenge on pulpal matrix metalloproteinase-13 (MMP-13) expression and elucidate the associated intracellular signalling pathways. Real-time PCR was used to detect the effects of CpG ODN on MMP-13 mRNA expression levels in a murine odontoblast-lineage cell line (OLCs). The possible involvement of TLR9/MyD88, NF-κB or MAPK pathways involved in the CpG ODN-induced MMP-13 expression was examined by real-time PCR, transient transfection, luciferase activity assay and ELISA. Western blotting was performed to assay the phosphorylation of ERK at a range of time points. MMP-13 was constitutively expressed in OLCs, and their exposure to CpG ODN significantly increased MMP-13 expression. Pre-treatment of OLCs with the inhibitory peptide MyD88, or chloroquine, attenuated the CpG ODN-induced expression of MMP-13. Treatment of the OLCs with CpG ODN increased NF-κB-luciferase activity. This activity was decreased by the over-expression of a nondegrading mutant of IκBα (IκBαSR), although enhanced by the over-expression of NF-κB p65. MMP-13 expression induced by CpG ODN was markedly suppressed by NF-κB inhibitors (pyrrolidine dithiocarbamate, PDTC), IκBα phosphorylation inhibitors (Bay 117082) or IκB protease inhibitor (L-1-tosylamido-2-phenylethyl chloromethyl ketone, TPCK). The inhibitor of ERK1/2, U0126, but not inhibitors of p38 MAPK and JNK, SB203580 and SP600125, decreased CpG ODN-mediated MMP-13 expression. The CpG ODN-induced MMP-13 expression in OLCs is mediated through TLR9, NF-κB and the ERK pathway indicating that potentially the recognition of CpG ODN by TLR9 on odontoblasts may regulate the remodelling of injured dental pulp and hard tissues by inducing MMP-13 expression.


WO3/g-C3N4 composites: one-pot preparation and enhanced photocatalytic H2 production under visible-light irradiation.
A series of WO3/g-C3N4 composites with different WO3 contents were prepared via a facile one-pot pyrolysis method, and showed notably enhanced visible-light-driven photocatalytic H2-evolution activities, with the highest rate of 400 μmol h-1 gcat-1 that was 15.0 times of that for pristine g-C3N4. Contents and sizes of WO3 crystallites in the composites were easily adjusted by changing the molar ratios of (NH4)2WS4 to C3H6N6 in the feed reagents, thereby successfully optimizing the Z-scheme system constructed by WO3 and g-C3N4 and thus effectively reducing the recombination of photogenerated charge carriers in g-C3N4. Moreover, pore volumes and surface areas of the composites were gradually enlarged by introducing WO3 into g-C3N4 via the one-pot preparation strategy, therefore promoting the redox reactions to evolve H2. This work presented an effective route to simultaneously optimize the phase compositions and textural structures of photocatalysts for enhanced H2 evolution.


Educational challenges faced by international medical graduates in the UK.
International medical graduates (IMGs) in the UK constitute approximately one-quarter of the total number of doctors registered in the General Medical Council (GMC). The transition of IMGs into the health care system in the UK is accompanied by significant sociocultural and educational challenges. This study aims to explore the views of IMGs in medical training on the educational challenges they face. This study was conducted in the Kent, Surrey and Sussex region in 2015. All IMGs who work in medical (physicianly) training programs were included. Data were collected through a questionnaire and semi-structured interviews. Thematic approach was used to analyze the qualitative data. Of the total 61 IMGs included, 17 responded to the survey and 3 were interviewed. The common educational barriers faced by IMGs were related to lack of appreciation of the values and structure of the National Health Service (NHS), ethical and medicolegal issues, receiving feedback from colleagues and the different learning strategies in the UK. IMGs suggested introduction of a mandatory dedicated induction program in the form of formal teaching sessions. They also believed that a supervised shadowing period prior in the first job in the UK would be beneficial. Further assessment areas should be incorporated into the prequalifying examinations to address specific educational needs such as NHS structure and hospital policies. Other measures such as buddying schemes with senior IMGs and educating NHS staff on different needs of IMGs should also be considered. This study highlighted important educational challenges faced by IMGs and generated relevant solutions. However, the opinions of the supervisors and other health care professionals need to be explored.


An examination of the living conditions of urban American Indian children in unmarried families: increasing cultural competence in child welfare.
The past 50 years have revealed dramatically shifting trends in the familial structure of American society. When examining these trends, and family research in general, the American Indian family unit has received little to no attention. This study utilized data from the Fragile Families and Child Wellbeing Study to examine the living conditions of urban American Indian children in unmarried families. Results showed that while these children appear to have a strong start, concerns are raised regarding American Indian mothers' low educational achievement and high incidence of poverty. These concerns can lead to potential issues regarding sustained development that can arise as the children grow. Therefore, child welfare workers must understand these issues and work to ameliorate them in order to provide culturally competent services to urban American Indian families and children.