Alpha-fetoprotein localization in pure ovarian teratoma. An immunohistochemical study of 12 cases.
Twelve cases of pure ovarian teratoma were studied by immunohistochemical methods to determine the incidence and localization of alpha-fetoprotein (AFP) production. There were six mature teratomas (grade 0) and six immature ones (grades 1-3). Three tumors (grades 1, 3, and 3 and stages III, I, and III, respectively) had isolated foci of AFP immunoreactivity in yolk-sac-like vesicles, intestinal-type epithelium, or tissue resembling liver, whereas the other nine tumors had no AFP-positive components. The former three patients had preoperative serum AFP determinations; the values were elevated in two of them, and in both instances the oncofetoantigen became undetectable one month after oophorectomy. Both patients are alive and well 15 and 25 months later; the third patient with a normal serum AFP before surgery and a positively stained tumor died of recurrent disease. The authors' findings indicate that the production of AFP in pure ovarian teratomas recapitulates its pattern of synthesis during ontogeny. Furthermore, it is suggested that a preoperative elevation of serum AFP in a pure ovarian teratoma does not seem to correlate with stage or grade, beyond minimizing the possibility that the tumor is grade 0, or mature.


Heterocyst and nitrogenase development in Anabaena cylindrica.
The differentiation of filaments of a continuous culture of Anabaena cylindrica after removal of fixed nitrogen has been examined. Rapid development of proheterocysts (4-5 h) and mature heterocysts (14 h) in an ordered sequence was observed; the development of the latter was concomitant with the onset of nitrogenase activity. Commitment times of 2-3 h (proheterocyst) and 5-0 (heterocyst) were measured. Evidence is presented that shows that heterocyst development, rather than any product of heterocyst function in nitrogen fixation, is responsible for the imposition of the pattern of heterocysts in a filament of vegative cells. Both phycocyanin content and carbon dioxide fixation declined markedly throughout the filament during the early stages of heterocyst development, indicating that all the cells of the filament are involved in the initial stages of the differentiation process. Heterocysts do not evolve oxygen in the light, but do respire. Nitrogenase activity in isolated heterocysts and in intact filaments was stimulated by phosphoenolpyruvate, ATP and dichlorophenolindophenol-ascorbate.


Potential Importance of Early Focal Radiotherapy Following Gross Total Resection for Long-Term Survival in Children With Embryonal Tumors With Multilayered Rosettes.
Embryonal tumor with multilayered rosettes (ETMR) is a rare, aggressive embryonal central nervous system tumor characterized by LIN28A expression and alterations in the C19MC locus. ETMRs predominantly occur in young children, have a dismal prognosis, and no definitive treatment guidelines have been established. We report on nine consecutive patients and review the role of initiation/timing of radiotherapy on survival. Between 2006 and 2018, nine patients were diagnosed with ETMR. Diagnosis was confirmed histopathologically, immunohistochemically and molecularly. Median age was 25 months (5-38). Location was supratentorial in five, pineal in three, and brainstem in one. Seven patients had a gross total resection, one a partial resection and one a biopsy at initial diagnosis. Chemotherapy augmented with intrathecal therapy started a median of 10 days (7-20) after surgery. Only two patients who after gross total resection received radiotherapy very early on (six weeks after diagnosis) are alive and in complete remission 56 and 50 months after diagnosis. All remaining patients for whom radiotherapy was deferred until the end of chemotherapy recurred, albeit none with leptomeningeal disease. A literature research identified 228 patients with ETMR. Including our patients only 26 (11%) of 237 patients survived >36 months with no evidence of disease at last follow-up. All but two long-term (>36 months) survivors received radiotherapy, ten of whom early on following gross total resection (GTR). GTR followed by early focal radiotherapy and intrathecal therapy to prevent leptomeningeal disease are potentially important to improve survival of ETMR in the absence of effective targeted therapies.


Immunolocalization of the Na+/Ca2+ exchanger in rabbit kidney.
We recently isolated a cDNA encoding a Na+/Ca2+ exchanger from rabbit kidney that was highly similar to the canine cardiac sarcolemmal Na+/Ca2+ exchanger. In the present study, we used two different antibodies to the exchanger to identify the protein and establish its cellular and subcellular localization in the kidney. The first antibody was prepared against a fusion protein consisting of 190 amino acids of the large, presumably intracellular loop of the rabbit renal exchanger fused to the maltose-binding protein. The second was a monoclonal antibody generated against the isolated purified canine cardiac sarcolemmal exchanger. To identify the Na+/Ca2+ exchanger protein, we performed immunoblot analysis against a membrane vesicle preparation from rabbit kidney cortex. Both antibodies immunoblotted proteins of 120 and 70 kDa that are known to be associated with the exchanger. Indirect immunofluorescence revealed that both antisera labeled the basolateral surface of the majority of cells in the connecting tubule (CNT). Since the phase-dense (intercalated) cells in the CNT were not stained, this suggested that the labeled cells were CNT cells. No labeling was detected in other nephron segments with the exception of occasional faint staining of the majority cell population of the cortical collecting duct. The fact that we did not detect labeling in other nephron segments is consistent with either 1) the absence of expression of the Na+/Ca2+ exchanger in these segments, 2) the expression of the exchanger in levels below the threshold of detection of the two antibodies used in this study, or 3) the exchanger in these segments is represented by a different isoform.(ABSTRACT TRUNCATED AT 250 WORDS)


Description of Propionispira arcuata sp. nov., isolated from a methanogenic reactor of cattle waste, reclassification of Zymophilus raffinosivorans and Zymophilus paucivorans as Propionispira raffinosivorans comb. nov. and Propionispira paucivorans comb. nov. and emended description of the genus Propionispira.
A strictly anaerobic bacterial strain, WK011(T), was isolated from a methanogenic reactor treating waste from cattle farms. The cells stained Gram-negative and were curved rods with a polar or subpolar flagellum. Spore formation was not observed. The optimum temperature for growth was 35 °C and the optimum pH was 6.7. Tests for oxidase, catalase and nitrate-reduction activities were negative. Hydrogen sulfide was produced. The strain fermented carbohydrates and produced acetate and propionate as major fermentation products. The genomic DNA G+C content was 41.7 mol%. The major cellular fatty acids were C15:0, C16:1ω9c and C18:1 dimethylacetal. The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid. The most closely related species to strain WK011(T) on the basis of 16S rRNA gene sequences were Propionispira arboris and Zymophilus raffinosivorans (95.6% sequence similarity to the type strains of both species). It was shown by phylogenetic and phenotypic examination of the type strains of related species, including the second species of the genus Zymophilus, Zymophilus paucivorans, that the two genera should be combined and that the two species of the genus Zymophilus should be transferred to the genus Propionispira, as Propionispira raffinosivorans comb. nov. (type strain SH2(T) = ATCC 49691(T) = DSM 20765(T)) and Propionispira paucivorans comb. nov. (type strain AA1(T) = ATCC 49689(T) = DSM 20756(T)), with an emended description of the genus Propionispira. Based on differences in the phylogenetic and phenotypic characteristics of strain WK011(T) from those of closely related species, the novel species Propionispira arcuata sp. nov. is proposed to accommodate the strain. The type strain is WK011(T) ( = JCM 16475(T) = DSM 22929(T)).


Chronic perineal pain related to sacral meningeal cysts.
Chronic perineal pain is an often encountered problem that is difficult to evaluate. Based on a series of 17 patients in whom urological, gynecological, and anorectal pathology was excluded, the authors compared magnetic resonance imaging (MRI) with computed tomographic (CT) scan with myelography in the investigation of chronic perineal pain. After a clinical neurological examination, patients underwent radiodiagnostic imaging of both techniques. Thirteen patients (76%) had one or more sacral meningeal cysts (MC) on MRI scan, whereas CT scan with myelography of the lumbar and sacral region revealed 7 patients (41%) with sacral MC. Sacral MC may be the etiology of chronic perineal pain in many instances, and MRI scan appears to be superior to CT scan with myelography in demonstrating sacral MC. Ten patients with sacral MC were operated on with moderate to excellent results 6 months after operation. Early postoperative results are encouraging, but further follow-up and larger series are required.


Hypomethylating Agent Therapy in Myelodysplastic Syndromes With Chromosome 3 Abnormalities.
Abnormalities of chromosome 3 in myelodysplastic syndromes (MDS), that is, inversion 3 (inv[3]), translocation 3q (t[3q]), or deletion 3q (del[3q]), are defined as poor-risk karyotypes in the Revised International Prognostic Scoring System (IPSS-R). The objective of this study was to further define the outcomes of patients with MDS with chromosome 3 abnormalities and address the impact of hypomethylating agent (HMA) therapy on this patient subset. Through the MDS Clinical Research Consortium, we identified 411 patients with chromosome 3 abnormalities and MDS or oligoblastic acute myeloid leukemia (20%-30% blasts). Specific chromosome 3 aberrations and cytogenetic complexity were predictive of survival; patients with t(3q) and isolated chromosome 3 had improved overall survival (OS), albeit still poor, whereas patients with complex cytogenetics, including those with 3p abnormalities, had inferior OS. Overall response rates to HMAs among this patient population were similar to those of patients with nonchromosome 3-MDS (52%, with a 25% complete remission rate), although with higher response rates in decitabine-treated patients (69% vs. 45%, P = .008). HMA therapy improved the OS of patients with higher-risk MDS compared with intensive chemotherapy (median OS of 15.5 vs. 8.2 months; P = .017). This improvement remained significant in multivariate analyses (hazard ratio, 0.60; P = .018); however, there were no chromosome 3 aberrations among this subgroup predictive of improved response rates to or survival from HMAs. Patients with MDS with chromosome 3 abnormalities represent a cytogenetic cohort with poor OS, and there is an urgent need for novel therapeutic strategies.


Thermoregulatory ability and mechanism do not differ consistently between neotropical and temperate butterflies.
Climate change is a major threat to species worldwide, yet it remains uncertain whether tropical or temperate species are more vulnerable to changing temperatures. To further our understanding of this, we used a standardised field protocol to (1) study the buffering ability (ability to regulate body temperature relative to surrounding air temperature) of neotropical (Panama) and temperate (the United Kingdom, Czech Republic and Austria) butterflies at the assemblage and family level, (2) determine if any differences in buffering ability were driven by morphological characteristics and (3) used ecologically relevant temperature measurements to investigate how butterflies use microclimates and behaviour to thermoregulate. We hypothesised that temperate butterflies would be better at buffering than neotropical butterflies as temperate species naturally experience a wider range of temperatures than their tropical counterparts. Contrary to our hypothesis, at the assemblage level, neotropical species (especially Nymphalidae) were better at buffering than temperate species, driven primarily by neotropical individuals cooling themselves more at higher air temperatures. Morphology was the main driver of differences in buffering ability between neotropical and temperate species as opposed to the thermal environment butterflies experienced. Temperate butterflies used postural thermoregulation to raise their body temperature more than neotropical butterflies, probably as an adaptation to temperate climates, but the selection of microclimates did not differ between regions. Our findings demonstrate that butterfly species have unique thermoregulatory strategies driven by behaviour and morphology, and that neotropical species are not likely to be more inherently vulnerable to warming than temperate species.


Zinc bioavailability in soybean meal.
A phytate-containing soy protein concentrate (SPC) diet (13.5 mg Zn/kg) and a phytate-free egg white diet (.3 mg Zn/kg) were used to determine the relative bioavailability (RBV) of Zn in dehulled soybean meal (SBM) based on Zn depletion-repletion growth bioassays in young chicks. After a 4-d Zn depletion period, chick weight gain responded linearly (P < .01) to graded increments of supplemental Zn (0 to 10 mg/kg) from ZnSO4 x 7H2O, whether added to the Zn-deficient SPC or egg white diet. Slope, however, was over twice as great for the standard curve relating weight gain to supplemental Zn intake for the egg white diet as for the SPC diet. Addition of 7 to 10 mg Zn/kg from SBM to either Zn-deficient diet increased (P < .01) weight gain, but a similar SBM addition to either diet made adequate in Zn did not increase weight gain. Using standard-curve methodology, RBV of Zn in SBM was 78% when the the SPC diet was used, but it was only 40% when the egg white diet was used. The phytate contained in the SPC basal diet therefore markedly reduced the efficiency of utilizing the supplemental inorganic Zn from ZnSO4 x 7H2O. This lowered the slope of the standard curve so that, on a relative basis, the Zn in SBM had a higher RBV value than was the case for Zn utilization in SBM with the phytate-free egg white diet. The 78% Zn RBV value in SBM would seem to have the greatest relevance for practical-type corn-SBM diets.


Contrasting Evolutionary Paths Among Indo-Pacific Pomacentrus Species Promoted by Extensive Pericentric Inversions and Genome Organization of Repetitive Sequences.
Pomacentrus (damselfishes) is one of the most characteristic groups of fishes in the Indo-Pacific coral reef. Its 77 described species exhibit a complex taxonomy with cryptic lineages across their extensive distribution. Periods of evolutionary divergences between them are very variable, and the cytogenetic events that followed their evolutionary diversification are largely unknown. In this respect, analyses of chromosomal divergence, within a phylogenetic perspective, are particularly informative regarding karyoevolutionary trends. As such, we conducted conventional cytogenetic and cytogenomic analyses in four Pomacentrus species (Pomacentrus similis, Pomacentrus auriventris, Pomacentrus moluccensis, and Pomacentrus cuneatus), through the mapping of repetitive DNA classes and transposable elements, including 18S rDNA, 5S rDNA, (CA)15, (GA)15, (CAA)10, Rex6, and U2 snDNA as markers. P. auriventris and P. similis, belonging to the Pomacentrus coelestis complex, have indistinguishable karyotypes (2n = 48; NF = 48), with a peculiar syntenic organization of ribosomal genes. On the other hand, P. moluccensis and P. cuneatus, belonging to another clade, exhibit very different karyotypes (2n = 48, NF = 86 and 92, respectively), with a large number of bi-armed chromosomes, where multiple pericentric inversions played a significant role in their karyotype organization. In this sense, different chromosomal pathways followed the phyletic diversification in the Pomacentrus genus, making possible the characterization of two well-contrasting species groups regarding their karyotype features. Despite this, pericentric inversions act as an effective postzygotic barrier in many organisms, which appear to be also the case for P. moluccensis and P. cuneatus; the extensive chromosomal similarities in the two species of P. coelestis complex suggest minor participation of chromosomal postzygotic barriers in the phyletic diversification of these species.


CXCR4-overexpressed exosomes from cardiosphere-derived cells attenuate myocardial ischemia/reperfusion injury by transferring miRNA to macrophages and regulating macrophage polarization.
Cardiosphere-derived cells (CDCs) are emerging as ideal candidates for managing cardiac inflammation, albeit with some limitations. Recent literatures have indicated that exosomes secreted by CDCs with C-X-C motif chemokine receptor 4 (CXCR4) overexpression can promote cardiac function after myocardial infarction and there have been some reports of miRNAs involved in ischemia/reperfusion (I/R) therapy. Therefore, we are interested in the role of CXCR4-overexpressed CDC-derived exosomes in delivering specific miRNA after myocardial I/R injury. In this research, we first constructed CDC-derived exosomes that overexpressed CXCR4 and miR-27a-5p, miR-182, or miR-101a. Then, we co-cultured the engineered exosomes with RAW264.7 cells and injected them intravenously into myocardial I/R model mice. In vitro, results showed that proinflammatory cytokines levels in the culture supernatant were decreased and the expression of M2 phenotypic markers were increased. Administration of engineered exosomes improved cardiac function, reduced infarct size, alleviated macrophage infiltration, and regulated M2 macrophage polarization after myocardial I/R, suggesting their implications in cardiac injury repair.


Sulconazole-Loaded Solid Lipid Nanoparticles for Enhanced Antifungal Activity: In Vitro and In Vivo Approach.
Solid lipid nanoparticles (SLNs) have the advantages of a cell-specific delivery and sustained release of hydrophobic drugs that can be exploited against infectious diseases. The topical delivery of hydrophobic drugs needs pharmaceutical strategies to enhance drug permeation, which is a challenge faced by conventional formulations containing a drug suspended in gel, creams or ointments. We report the fabrication and optimization of SLNs with sulconazole (SCZ) as a model hydrophobic drug and then a formulation of an SLN-based topical gel against fungal infections. The SLNs were optimized through excipients of glyceryl monostearate and Phospholipon® 90 H as lipids and tween 20 as a surfactant for its size, drug entrapment and sustained release and resistance against aggregation. The SCZ-SLNs were physically characterized for their particle size (89.81 ± 2.64), polydispersity index (0.311 ± 0.07), zeta potential (-26.98 ± 1.19) and encapsulation efficiency (86.52 ± 0.53). The SCZ-SLNs showed sustained release of 85.29% drug at the 12 h timepoint. The TEM results demonstrated spherical morphology, while DSC, XRD and FTIR showed the compatibility of the drug inside SLNs. SCZ-SLNs were incorporated into a gel using carbopol and were further optimized for their rheological behavior, pH, homogeneity and spreadability on the skin. The antifungal activity against Candida albicans and Trichophyton rubrum was increased in comparison to a SCZ carbopol-based gel. In vivo antifungal activity in rabbits presented faster healing of skin fungal infections. The histopathological examination of the treated skin from rabbits presented restoration of the dermal architecture. In summary, the approach of formulating SLNs into a topical gel presented an advantageous drug delivery system against mycosis.


The Rostro-Caudal Axis of Frontal Cortex Is Sensitive to the Domain of Stimulus Information.
Evidence suggests that lateral frontal cortex implements cognitive control processing along its rostro-caudal axis, yet other evidence supports a dorsal-ventral functional organization for processes engaged by different stimulus domains (e.g., spatial vs. nonspatial). This functional magnetic resonance imaging study investigated whether separable dorsolateral and ventrolateral rostro-caudal gradients exist in humans, while participants performed tasks requiring cognitive control at 3 levels of abstraction with language or spatial stimuli. Abstraction was manipulated by using 3 different task sets that varied in relational complexity. Relational complexity refers to the process of manipulating the relationship between task components (e.g., to associate a particular cue with a task) and drawing inferences about that relationship. Tasks using different stimulus domains engaged distinct posterior regions, but within the lateral frontal cortex, we found evidence for a single rostro-caudal gradient that was organized according to the level of abstraction and was independent of processing of the stimulus domain. However, a pattern of dorsal/ventral segregation of processing engaged by domain-specific information was evident in each separable frontal region only within the most rostral region recruited by task demands. These results suggest that increasingly abstract information is represented in the frontal cortex along distinct rostro-caudal gradients that also segregate along a dorsal-ventral axis dependent on task demands.


Cholecystitis and cholelithiasis masking as abdominal crises in sickle cell disease.
A case of sickle cell (HbSS) disease is presented in a patient with a history of recurrent admissions for abdominal pain, jaundice, and abnormal liver function tests. Although he was believed to have a sickle cell abdominal crisis, his abdominal X-ray films revealed three calcified stones. Each of these stones progressively passed through the common duct and into the duodenum while awaiting surgery. He has been followed for two years since his cholecystectomy without further hospitalizations. This case led to the investigation of cholelithiasis in sickle cell disease to dispel the following misconceptions. Some physicians and pediatricians believe that (1) cholelithiasis and cholecystitis are uncommon in sickle cell disease; (2) the complications of gallstones are not significant; (3) the operative risk in patients with sickle cell disease is high; (4) these patients with HbSS disease do not live long enough to get into trouble with gallstones. A review of the literature on cholelithiasis and HbSS disease presents adequate evidence to cause us to urge investigation of the gallbladder in all patients with HbSS disease and abdominal crises, and cholecystectomy as an elective procedure should stones be present.


Thermoelectric performance of nanostructured In/Pb codoped SnTe with band convergence and resonant level prepared via a green and facile hydrothermal method.
SnTe is considered as a promising alternative to the conventional high-performance thermoelectric material PbTe, which inspired the thermoelectric community for a while. Here, we design a green, facile and low-energy-intensity hydrothermal route without involving any toxic or unstable chemicals to fabricate SnTe-based thermoelectric materials. Ultralow lattice thermal conductivity and enhanced thermoelectric performance are achieved via the combination of band engineering and nanostructuring. Enhanced Seebeck coefficient and power factor are induced by converging the band structure and creating resonant levels due to Pb and In doping. More importantly, due to the reduced grain sizes, nanoparticles, and dual-atom point defect scattering, ultralow lattice thermal conductivity was obtained in the bulk samples fabricated by the hydrothermal route. Benefiting from the enhanced power factor and significantly reduced thermal conductivity, the peak ZT is enhanced to ∼0.7 in In/Pb codoped SnTe, a 60% improvement over pure SnTe.


Inositol 1,3,4,5-tetrakisphosphate and not phosphatidylinositol 3,4-bisphosphate is the probable precursor of inositol 1,3,4-trisphosphate in agonist-stimulated parotid gland.
When [3H]inositol-prelabelled rat parotid-gland slices were stimulated with carbachol, noradrenaline or Substance P, the major inositol trisphosphate produced with prolonged exposure to agonists was, in each case, inositol 1,3,4-trisphosphate. Much lower amounts of radioactivity were present in the inositol 1,4,5-trisphosphate fraction separated by anion-exchange h.p.l.c. Analysis of the inositol trisphosphate head group of phosphatidylinositol bisphosphate in [32P]Pi-labelled parotid glands showed the presence of phosphatidylinositol 4,5-bisphosphate, but no detectable phosphatidylinositol 3,4-bisphosphate. Carbachol-stimulated [3H]inositol-labelled parotid glands contained an inositol polyphosphate with the chromatographic properties and electrophoretic mobility of an inositol tetrakisphosphate, the probable structure of which was determined to be inositol 1,3,4,5-tetrakisphosphate. Since an enzyme in erythrocyte membranes is capable of degrading this tetrakisphosphate to inositol 1,3,4-trisphosphate, it is suggested to be the precursor of inositol 1,3,4-trisphosphate in parotid glands.


Sheep persistently infected with Border disease readily transmit virus to calves seronegative to BVD virus.
Bovine viral diarrhea- and Border disease viruses of sheep belong to the highly diverse genus pestivirus of the Flaviviridae. Ruminant pestiviruses may infect a wide range of domestic and wild cloven-hooved mammals (artiodactyla). Due to its economic importance, programs to eradicate bovine viral diarrhea are a high priority in the cattle industry. By contrast, Border disease is not a target of eradication, although the Border disease virus is known to be capable of also infecting cattle. In this work, we compared single dose experimental inoculation of calves with Border disease virus with co-mingling of calves with sheep persistently infected with this virus. As indicated by seroconversion, infection was achieved only in one out of seven calves with a dose of Border disease virus that was previously shown to be successful in calves inoculated with BVD virus. By contrast, all calves kept together with persistently infected sheep readily became infected with Border disease virus. The ease of viral transmission from sheep to cattle and the antigenic similarity of bovine and ovine pestiviruses may become a problem for demonstrating freedom of BVD by serology in the cattle population.


Nationwide claims database analysis of treatment patterns, costs and survival of Japanese patients with diffuse large B-cell lymphoma.
Limited data are available regarding treatment patterns, healthcare resource utilization (HCRU), treatment costs and clinical outcomes for patients with diffuse large B-cell lymphoma (DLBCL) in Japan. This retrospective database study analyzed the Medical Data Vision database for DLBCL patients who received treatment during the identification period from October 1 2008 to December 31 2017. Among 6,965 eligible DLBCL patients, 5,541 patients (79.6%) received first-line (1L) rituximab (R)-based therapy, and then were gradually switched to chemotherapy without R in subsequent lines of therapy. In each treatment regimen, 1L treatment cost was the highest among all lines of therapy. The major cost drivers i.e. total direct medical costs until death or censoring across all regimens and lines of therapy were from the 1L regimen and inpatient costs. During the follow-up period, DLBCL patients who received a 1L R-CHOP regimen achieved the highest survival rate and longest time-to-next-treatment, with a relatively low mean treatment cost due to lower inpatient healthcare resource utilization and fewer lines of therapy compared to other 1L regimens. Our retrospective analysis of clinical practices in Japanese DLBCL patients demonstrated that 1L treatment and inpatient costs were major cost contributors and that the use of 1L R-CHOP was associated with better clinical outcomes at a relatively low mean treatment cost.


Nuclear import of HIV-1 DNA in resting CD4+ T cells requires a cyclosporin A-sensitive pathway.
Using PCR to monitor early (LTR/LTR (long terminal repeat)) and late (LTR/gag) products of reverse transcription and the formation of HIV-1 LTR circles (indicating nuclear import), we explored the relationship between T cell activation signals and early events in the life cycle of HIV-1 infection. The combination of TCR ligation with either CD28 cross-linking or exogenous IL-2 was required for HIV-1 LTR circle formation in resting CD4+ T cells. Ligation of the TCR or CD28 receptors or addition of IL-2 alone did not induce this process. However, cross-linking the TCR, or IL-2 alone, unlike CD28 ligation, could induce the completion of viral reverse transcription. In contrast, the initiation of HIV-1 reverse transcription could occur in resting CD4+ T cells without any stimulation. Cyclosporin A (CsA), an inhibitor of T cell activation, completely blocked HIV-1 DNA nuclear import in activated CD4+ T cells. The completion of HIV-1 reverse transcription was blocked by CsA in infected CD4+ T cells activated by TCR ligation and IL-2, but not in cells stimulated by TCR and CD28 ligation. The costimulation of CD3 and CD28 mAbs in the presence of IL-2 could not overcome the CsA inhibitory effect on nuclear import of viral DNA. Therefore, the factor(s) involved in a CsA-sensitive pathway plays a critical role in HIV-1 DNA transport from the cytoplasm into the nucleus. Production and movement of HIV-1 DNA in resting CD4+ T cells require two signals: one signal from the TCR, which normally regulates the G0 to G1 transition, induces completion of viral reverse transcription; the other signal through CD28 or an IL-2R-dependent process is sensitive to CsA treatment and regulates viral DNA entry into the nucleus.


The neuropeptide Y Y1 receptor mediates NPY-induced inhibition of the gonadotrope axis under poor metabolic conditions.
Hypothalamic neuropeptide Y (NPY) plays a central role in the control of food intake, energy balance, and modulation of neuroendocrine functions. In particular, an increase in NPY expression participates in the inhibition of the reproductive activity under poor nutritional conditions. The present study was designed to evaluate further the involvement of the Y1 subtype of NPY receptors in these effects. Food intake, body weight gain, and the onset of puberty were studied in groups of wild-type and Y1 deficient mice that were either fed ad libitum or subjected to a 30% restriction in food intake. This moderate feeding restriction induced a similar deficit in body weight gain in wild-type and in Y1 knockout mice. However, although wild-type mice experienced the expected delay of puberty, all mice in the food restriction group and lacking Y1 could go through puberty over the time of the experiment despite decreases in circulating leptin levels and increases in hypothalamic NPY expression. This observation demonstrates that the absence of Y1 impairs the perception of decreasing energy stores by the gonadotrope axis, demonstrating a physiological role for Y1 in the sensing of endogenous metabolic parameters by the hypothalamus.


[Less refusal to participate in HIV screening among pregnant women in the Amsterdam region since the introduction of standard HIV screening using the opting-out method].
In 2003 the Municipal Health Service in Amsterdam started to screen pregnant women for HIV according to the opting-out method. In this method the HIV test is routinely included in the prenatal screening along with hepatitis B virus (HBV) and syphilis. If the woman does not want to be tested for HIV then she must actively opt out of this test. This screening method was chosen because in the universal screening method used in 2002, women had to give their explicit consent to test for HIV and this led to a high refusal rate (13.6%), especially among women from AIDS-endemic countries. After the introduction of the opting-out method, the refusal rate fell from 3% in the first quarter of 2003 to 1.4% in the last quarter of 2003. None of the women refused to be tested for HBV or syphilis. In 2003, the HIV prevalence among pregnant women was 0.3% (35/13.621). The experiences with this screening method in Amsterdam were used to implement the national opting-out method for HIV screening in pregnant women, which was introduced on 1 January 2004.


The Relationship between auxin transport and maize branching.
Maize (Zea mays) plants make different types of vegetative or reproductive branches during development. Branches develop from axillary meristems produced on the flanks of the vegetative or inflorescence shoot apical meristem. Among these branches are the spikelets, short grass-specific structures, produced by determinate axillary spikelet-pair and spikelet meristems. We investigated the mechanism of branching in maize by making transgenic plants expressing a native expressed endogenous auxin efflux transporter (ZmPIN1a) fused to yellow fluorescent protein and a synthetic auxin-responsive promoter (DR5rev) driving red fluorescent protein. By imaging these plants, we found that all maize branching events during vegetative and reproductive development appear to be regulated by the creation of auxin response maxima through the activity of polar auxin transporters. We also found that the auxin transporter ZmPIN1a is functional, as it can rescue the polar auxin transport defects of the Arabidopsis (Arabidopsis thaliana) pin1-3 mutant. Based on this and on the groundbreaking analysis in Arabidopsis and other species, we conclude that branching mechanisms are conserved and can, in addition, explain the formation of axillary meristems (spikelet-pair and spikelet meristems) that are unique to grasses. We also found that BARREN STALK1 is required for the creation of auxin response maxima at the flanks of the inflorescence meristem, suggesting a role in the initiation of polar auxin transport for axillary meristem formation. Based on our results, we propose a general model for branching during maize inflorescence development.


Inter-rater reliability of data elements from a prototype of the Paul Coverdell National Acute Stroke Registry.
The Paul Coverdell National Acute Stroke Registry (PCNASR) is a U.S. based national registry designed to monitor and improve the quality of acute stroke care delivered by hospitals. The registry monitors care through specific performance measures, the accuracy of which depends in part on the reliability of the individual data elements used to construct them. This study describes the inter-rater reliability of data elements collected in Michigan's state-based prototype of the PCNASR. Over a 6-month period, 15 hospitals participating in the Michigan PCNASR prototype submitted data on 2566 acute stroke admissions. Trained hospital staff prospectively identified acute stroke admissions, abstracted chart information, and submitted data to the registry. At each hospital 8 randomly selected cases were re-abstracted by an experienced research nurse. Inter-rater reliability was estimated by the kappa statistic for nominal variables, and intraclass correlation coefficient (ICC) for ordinal and continuous variables. Factors that can negatively impact the kappa statistic (i.e., trait prevalence and rater bias) were also evaluated. A total of 104 charts were available for re-abstraction. Excellent reliability (kappa or ICC > 0.75) was observed for many registry variables including age, gender, black race, hemorrhagic stroke, discharge medications, and modified Rankin Score. Agreement was at least moderate (i.e., 0.75 > kappa >/=; 0.40) for ischemic stroke, TIA, white race, non-ambulance arrival, hospital transfer and direct admit. However, several variables had poor reliability (kappa < 0.40) including stroke onset time, stroke team consultation, time of initial brain imaging, and discharge destination. There were marked systematic differences between hospital abstractors and the audit abstractor (i.e., rater bias) for many of the data elements recorded in the emergency department. The excellent reliability of many of the data elements supports the use of the PCNASR to monitor and improve care. However, the poor reliability for several variables, particularly time-related events in the emergency department, indicates the need for concerted efforts to improve the quality of data collection. Specific recommendations include improvements to data definitions, abstractor training, and the development of ED-based real-time data collection systems.


Treatment with Ibrutinib Inhibits BTK- and VLA-4-Dependent Adhesion of Chronic Lymphocytic Leukemia Cells In Vivo.
Ibrutinib leads to a transient lymphocytosis in patients with chronic lymphocytic leukemia (CLL) that develops within hours of starting drug and is due to the efflux of cells from lymphoid tissues into the blood. We therefore sought to investigate the in vivo effect of ibrutinib on migration and adhesion of CLL cells. Patients received single-agent ibrutinib (420 mg daily) on an investigator-initiated phase II trial. Serial blood samples were collected pretreatment and during treatment for ex vivo functional assays. Adhesion of CLL cells to fibronectin was rapidly (within hours) and almost completely inhibited (median reduction 98% on day 28, P < 0.001), while the effect on migration to chemokines was more moderate (median reduction 64%, P = 0.008) and less uniform. Although cell surface expression of key adhesion molecules such as CD49d, CD29, and CD44 were modestly reduced, this was only apparent after weeks of treatment. Stimulation of CLL cells from patients on ibrutinib with PMA, which activates PKC independent of BTK, restored the ability of the cells to adhere to fibronectin in a VLA-4-dependent manner. Finally, the addition of ibrutinib to CLL cells adhered to fibronectin in vitro caused the detachment of 17% of the cells, on average; consisten t with in vivo observations of an increasing lymphocytosis within 4 hours of starting ibrutinib. Inhibition of BTK and VLA-4-dependent adhesion of CLL cells to stroma and stromal components provides a mechanistic explanation for the treatment-induced lymphocytosis and may reduce CD49d-dependent prosurvival signals in the tissue microenvironment.


28W average power hydrocarbon-free rubidium diode pumped alkali laser.
We present experimental results for a high-power diode pumped hydrocarbon-free rubidium laser with a scalable architecture. The laser consists of a liquid cooled, copper waveguide which serves to both guide the pump light and to provide a thermally conductive surface near the gain volume to remove heat. A laser diode stack, with a linewidth narrowed to approximately 0.35 nm with volume bragg gratings, is used to pump the cell. We have achieved 24W average power output using 4 atmospheres of naturally occurring helium ((4)He) as the buffer gas and 28W using 2.8 atmospheres of (3)He.


High doses of vitamin E in the treatment of disorders of the central nervous system in the aged.
Oxidative stress is a putative factor in the pathogenesis of many human disorders of the central nervous system. Therefore, antioxidants such as vitamin E have become attractive as therapeutic agents in the treatment of several diseases. In addition, vitamin E seems to play a specific role in the nervous system. As a result, vitamin E has been used in pharmacologic doses in the treatment of disorders such as Parkinson disease, Alzheimer disease, and tardive dyskinesia. One investigation showed that the use of 2000 IU all-rac-alpha-tocopheryl acetate is beneficial in the treatment of Alzheimer disease. Similar doses of vitamin E, however, were not beneficial for delaying the progression of Parkinson disease. In other studies, dosages >/=400 IU vitamin E/d were found to be beneficial in the treatment of tardive dyskinesia, although this finding was not confirmed in a larger cooperative study conducted by the Veterans Administration. Even though the efficacy of vitamin E in the management of cardiovascular disease has been shown, the potential role of vitamin E in the treatment of cerebrovascular disease remains essentially unknown. The experience from 2 large clinical trials involving the oral intake of 2000 IU vitamin E/d suggests that vitamin E is relatively safe at this dosage for periods <2 y. However, the safety and efficacy of supplemental vitamin E over periods of many years in the prevention of neurologic diseases has not been adequately explored.


Human apolipoprotein E expression from mouse skeletal muscle by electrotransfer of nonviral DNA (plasmid) and pseudotyped recombinant adeno-associated virus (AAV2/7).
Plasma apolipoprotein E (apoE) has multiple atheroprotective actions. However, although liver-directed adenoviral gene transfer of apoE reverses hypercholesterolemia and inhibits atherogenesis in apoE-deficient (apoE(-/-)) mice, safety considerations have revived interest in nonviral DNA (plasmid) and nonpathogenic adeno-associated viral (AAV) vectors. Here, we assess the effectiveness of these two delivery vehicles by minimally invasive intramuscular injection. First, we constructed AAV2-based expression plasmids harboring human apoE3 cDNA, driven by two muscle-specific promoters (CK6 and C5-12) and one ubiquitous promoter (CAG); each efficiently expressed apoE3 in transfected cultured C2C12 mouse myoblasts, although muscle-specific promoters were active only in differentiated multinucleate myotubes. Second, a pilot study verified that electrotransfer of the CAG-driven plasmid (p.CAG.apoE3) into tibialis anterior muscles, pretreated with hyaluronidase, of apoE(-/-) mice significantly enhanced (p < 0.001) local intramuscular expression of apoE3. However, in a 7-day experiment, the CK6- and C5-12-driven plasmids produced less apoE3 in muscle than did p.CAG.apoE3 (0.61 +/- 0.38 and 0.45 +/- 0.38 vs. 13.38 +/- 7.46 microg of apoE3 per muscle, respectively), but plasma apoE3 levels were below our detection limit (<15 ng/ml) in all mice and did not reverse the hyperlipidemia. Finally, we showed that intramuscular injection of a cross-packaged AAV serotype 7 viral vector, expressing human apoE3 from the CAG promoter, resulted in increasing levels of apoE3 in plasma over 4 weeks, although the concentration reached (1.40 +/- 0.35 microg/ml) was just below the threshold level needed to reduce the hypercholesterolemia. We conclude that skeletal muscle can serve as an effective secretory platform to express the apoE3 transgene, but that improved gene transfer vectors are needed to achieve full therapeutic levels of plasma apoE3 protein.


The effect of talking about psychological trauma with a significant other on heart rate reactivity in individuals with posttraumatic stress disorder.
Individuals with posttraumatic stress disorder (PTSD) commonly make efforts to avoid trauma-oriented conversations with their significant others, which may interfere with the natural recovery process. Trauma-oriented conversations can be experienced as physiologically arousing, depending on the intensity of PTSD symptoms and perceptions of social support. In the current investigation, changes in heart rate responses to a trauma-oriented social interaction with a significant other were assessed. Perceived supportive and unsupportive or negative social interactions were examined as moderators of the association between heart rate changes to this context and intensity of PTSD symptoms. A total of 46 individuals with PTSD completed diagnostic interviews and self-report measures of symptoms and perceived supportive and negative social interactions during a trauma-oriented social interaction with a significant other. Heart rate was continuously measured during this interaction. Results showed that engagement in a trauma-oriented social interaction was predictive of elevations in heart rate that positively correlated with intensity of PTSD symptoms. The moderation hypothesis was partially supported. In addition, perceived negative social interactions positively correlated with elevations in heart rate. These findings can inform social intervention efforts for individuals with PTSD.


Mixture models for analysis of melting temperature data.
In addition to their use in detecting undesired real-time PCR products, melting temperatures are useful for detecting variations in the desired target sequences. Methodological improvements in recent years allow the generation of high-resolution melting-temperature (Tm) data. However, there is currently no convention on how to statistically analyze such high-resolution Tm data. Mixture model analysis was applied to Tm data. Models were selected based on Akaike's information criterion. Mixture model analysis correctly identified categories in Tm data obtained for known plasmid targets. Using simulated data, we investigated the number of observations required for model construction. The precision of the reported mixing proportions from data fitted to a preconstructed model was also evaluated. Mixture model analysis of Tm data allows the minimum number of different sequences in a set of amplicons and their relative frequencies to be determined. This approach allows Tm data to be analyzed, classified, and compared in an unbiased manner.


High efficiency silicon nanowire/organic hybrid solar cells with two-step surface treatment.
A simple two-step surface treatment process is proposed to boost the efficiency of silicon nanowire/PEDOT:PSS hybrid solar cells. The Si nanowires (SiNWs) are first subjected to a low temperature ozone treatment to form a surface sacrificial oxide, followed by a HF etching process to partially remove the oxide. TEM investigation demonstrates that a clean SiNW surface is achieved after the treatment, in contrast to untreated SiNWs that have Ag nanoparticles left on the surface from the metal-catalyzed etching process that is used to form the SiNWs. The cleaner SiNW surface achieved and the thin layer of residual SiO2 on the SiNWs have been found to improve the performance of the hybrid solar cells. Overall, the surface recombination of the hybrid SiNW solar cells is greatly suppressed, resulting in a remarkably improved open circuit voltage of 0.58 V. The power conversion efficiency has also increased from about 10% to 12.4%. The two-step surface treatment method is promising in enhancing the photovoltaic performance of the hybrid silicon solar cells, and can also be applied to other silicon nanostructure based solar cells.


Volume independence in the large N limit and an emergent fermionic symmetry.
Large-N volume independence in circle-compactified QCD with adjoint Weyl fermions implies the absence of any phase transitions as the radius is dialed to arbitrarily small values. This class of theories is believed to possess a Hagedorn density of hadronic states. It turns out that these properties are in apparent tension with each other, because a Hagedorn density of states typically implies a phase transition at some finite radius. This tension is resolved if there are degeneracies between the spectra of bosonic and fermionic states, as happens in the N(f) = 1 supersymmetric case. Resolution of the tension for N(f) >1 then suggests the emergence of a fermionic symmetry at large N, where there is no supersymmetry. We can escape the Coleman-Mandula theorem since the N = ∞ theory is free, with a trivial S matrix. We show an example of such a spectral degeneracy in a nonsupersymmetric toy example which has a Hagedorn spectrum.


Induction of dark-adaptive retinomotor movement (cell elongation) in teleost retinal cones by cyclic adenosine 3','5-monophosphate.
In the teleost retina, the photoreceptors and retinal pigment epithelium (RPE) undergo extensive movements (called retinomotor movements) in response to changes in light conditions and to an endogenous circadian rhythm. Photoreceptor movements serve to reposition the light-receptive outer segments and are effected by changes in inner segment length. Melanin granule movements within the RPE cells provide a movable melanin screen for rod outer segments. In the dark (night), cones elongate, rods contract, and pigment granules aggregate to the base of the RPE cell; in the light (day), these movements are reversed. We report here that treatments that elevate cytoplasmic cyclic adenosine 3',5'-monophosphate (cAMP) provoke retinomotor movements characteristic of nighttime dark adaptation, even in bright light at midday. To illustrate this response, we present a quantitative description of the effects of cyclic nucleotides on cone length in the green sunfish, Lepomis cyanellus. Cone elongation is induced when light-adapted retinas are exposed to exogenous cAMP analogues accompanied by phosphodiesterase (PDE) inhibitors (either by intraocular injection or in retinal organ culture). Cone movements is not affected by cyclic GMP analogies. Dose-response studies indicate that the extent, but not the rate, of cone elongation is proportional to the concentration of exogenous cAMP and analogue presented. As has been reported for other species, we find that levels of cAMP are significantly higher in dark- than in light-adapted green sunfish retinas. On the basis of these observations, we suggest that cAMP plays a role in the light and circadian regulation of teleost cone length.


Kaposi's sarcoma-associated herpesvirus-infected primary effusion lymphoma has a plasma cell gene expression profile.
Kaposi's sarcoma-associated herpesvirus is associated with three human tumors: Kaposi's sarcoma, and the B cell lymphomas, plasmablastic lymphoma associated with multicentric Castleman's disease, and primary effusion lymphoma (PEL). Epstein-Barr virus, the closest human relative of Kaposi's sarcoma-associated herpesvirus, mimics host B cell signaling pathways to direct B cell development toward a memory B cell phenotype. Epstein-Barr virus-associated B cell tumors are presumed to arise as a consequence of this virus-mediated B cell activation. The stage of B cell development represented by PEL, how this stage relates to tumor pathology, and how this information may be used to treat the disease are largely unknown. In this study we used gene expression profiling to order a range of B cell tumors by stage of development. PEL gene expression closely resembles that of malignant plasma cells, including the low expression of mature B cell genes. The unfolded protein response is partially activated in PEL, but is fully activated in plasma cell tumors, linking endoplasmic reticulum stress to plasma cell development through XBP-1. PEL cells can be defined by the overexpression of genes involved in inflammation, cell adhesion, and invasion, which may be responsible for their presentation in body cavities. Similar to malignant plasma cells, all PEL samples tested express the vitamin D receptor and are sensitive to the vitamin D analogue drug EB 1089 (Seocalcitol).


Decreased imipramine binding in the brains of patients with depressive illness.
The binding of tritiated imipramine was significantly reduced in the hippocampus and occipital cortex from a series of patients with depressive illness compared with age-matched patients with no psychiatric disorder. In contrast there was no change in imipramine binding in established cases of senile dementia of Alzheimer-type. Scatchard analysis indicated normal binding affinity but a reduction in the number of imipramine binding sites in depression. These observations parallel previous findings of decreased binding sites in platelets from depressed patients and suggest there may be an abnormality in the uptake mechanism for serotonin in depression.


Bismuth Oxychalcogenides: A New Class of Ferroelectric/Ferroelastic Materials with Ultra High Mobility.
Atomically thin Bi2O2Se has been recently synthesized, and it possesses ultrahigh mobility (Nat. Nanotechnol. 2017, 12, 530; Nano Lett. 2017, 17, 3021). Herein, we show first-principles evidence that Bi2O2Se and a related class of bismuth oxychalcogenides, such as Bi2O2S and Bi2O2Te, not only are novel semiconductors with ultrahigh mobility but also possess previously unreported ferroelectricity/ferroelasticity. Such a unique combination of semiconducting with ferroelectric/ferroelastic properties enables bismuth oxychalcogenides to potentially meet a great challenge, that is, integration of room-temperature functional nonvolatile memories into future nanocircuits. Specifically, we predict that bulk Bi2O2S is both ferroelastic and antiferroelectric and that a thin film with odd number of layers can even be multiferroic with nonzero in-plane polarization, and this polarization can be switchable via ferroelasticity. Moreover, Bi2O2Te possesses intrinsic out-of-plane ferroelectricity, while Bi2O2Se possesses piezoelectricity and ferroelectricity upon an in-plane strain. The in-plane strain on Bi2O2Se can induce giant polarizations (56.1 μC/cm2 upon 4.1% strain) with the piezoelectric coefficient being about 35 times higher than that of MoS2 monolayer. The in-plane strain can also enhance the bandgap or even convert indirect to direct bandgap beyond a critical value. The good match among the lattice constants of bismuth oxychalcogenides is also desirable, rendering the epitaxial growth of heterostructure devices free of fabrication issues related to lattice mismatch, thereby allowing high-quality bismuth oxychalcogenide heterostructures tailored by design for a variety of applications.


Influence of mast cells on thyroid function.
The role of mast cells on thyroid gland function was studied with respect to two sites of thyroid control: 1. the thyroid gland; 2. the hypothalamus pituitary system. It was observed that the treatment with disodium cromoglycate (mast cells stabilizer) partially blocked the TSH actions on thyroid gland, thus evidencing the thyroidal mast cells as an intermediary factor of TSH actions on thyroid gland. Furthermore, both T4 and T3 serum concentrations were increased after compound BW48-80 (mast cells disrupter polymer) was administered, concomitantly with increased 1 and 24 h radioiodine uptake, T/S ratio, PB131I and TSH serum concentration. The alterations were not observed in the rats pre-treated with disodium cromoglycate and administered with BW48-80. Finally, BW48-80 promoted an elevation of TSH serum concentration in T4-primed thyroidectomized rats. This phenomenon was completely prevented by the pretreatment with cromoglycate, which confirmed the active participation of mast cells on TRH and/or TSH secretion.


Alteration of NMDA NR1 receptors within the paraventricular nucleus of hypothalamus in rats with heart failure.
One of the pathophysiological characteristics of chronic heart failure (HF) is elevated sympathetic drive, which is a major factor contributing to the morbidity and mortality of HF. Resent evidence points to a central mechanism that contributes to the sympathetic abnormality in HF. The paraventricular nucleus (PVN) of the hypothalamus is an important site that integrates sympathetic nerve activity. Studies have shown that glutamate elicits excitatory effects on neurons in the PVN through the NMDA receptor. The goal of the present study was to examine the role of NMDA receptors in the altered sympathetic nerve activation during HF. The left coronary ligation-induced heart failure model in the rat was used. In alpha-chloralose and urethane anesthetized rats, microinjection of NMDA into the PVN (50 to 200 pmol) produced dose-dependent increases in renal sympathetic nerve discharge (RSND), arterial blood pressure (BP), and heart rate (HR). This response to NMDA was significantly potentiated (27+/-7%) in HF compared with sham rats. On the other hand, microinjection of the NMDA receptor antagonist AP-5 (4 to 16 nmol) into the PVN caused significant decreases in RSND, BP, and HR only in rats with HF but very slight changes in sham rats. Furthermore, using microdialysis and HPLC in combination with electrochemical detection techniques, we found that the glutamate level in the PVN was not increased significantly in HF compared with sham rats. However, using RT-PCR, Western blot, and immunofluorescence techniques, it was found that NMDA NR1 subunit mRNA expression and protein level in the PVN were significantly increased in HF compared with sham rats. These data suggest that the increased glutamatergic activity on sympathetic regulation, due to the upregulation of NMDA NR1 receptor subunits within the PVN may contribute to the elevated sympathoexcitation during HF.


Cooperative inhibition of T-cell antigen receptor signaling by a complex between a kinase and a phosphatase.
Antigen receptor-triggered T-cell activation is mediated by the sequential action of the Src and Syk/Zap-70 families of protein tyrosine kinases (PTKs). Previously, we reported that another PTK termed p50(csk) was a potent negative regulator of T-cell receptor (TCR) signaling because of its ability to inactivate Src-related kinases. This inhibitory effect required the catalytic activity of Csk, as well as its Src homology (SH)3 and SH2 domains. Subsequent studies uncovered that, via its SH3 domain, p50(csk) was associated with PEP, a proline-enriched protein tyrosine phosphatase (PTP) of unknown function expressed in hemopoietic cells. Herein, we have attempted to identify the role of the Csk-PEP complex in T lymphocytes. The results of our experiments showed that, like Csk, PEP was a strong repressor of TCR signaling. This property was dependent on the phosphatase activity of PEP, as well as on the sequence mediating its binding to p50(csk). Through reconstitution experiments in Cos-1 cells, evidence was obtained that Csk and PEP act synergistically to inhibit protein tyrosine phosphorylation by Src-related kinases, and that this effect requires their association. Finally, experiments with a substrate-trapping mutant of PEP suggested that PEP functions by dephosphorylating and inactivating the PTKs responsible for T-cell activation. In addition to giving novel insights into the mechanisms involved in the negative regulation of T-cell activation, these findings indicate that the association of an inhibitory PTK with a PTP constitutes a more efficient means of inhibiting signal transduction by Src family kinases in vivo.


Emotional Abilities in Children with Oppositional Defiant Disorder (ODD): Impairments in Perspective-Taking and Understanding Mixed Emotions are Associated with High Callous-Unemotional Traits.
Most studies of emotion abilities in disruptive children focus on emotion expression recognition. This study compared 74 children aged 4-8 years with ODD to 45 comparison children (33 healthy; 12 with an anxiety disorder) on behaviourally assessed measures of emotion perception, emotion perspective-taking, knowledge of emotions causes and understanding ambivalent emotions and on parent-reported cognitive and affective empathy. Adjusting for child's sex, age and expressive language ODD children showed a paucity in attributing causes to emotions but no other deficits relative to the comparison groups. ODD boys with high levels of callous-unemotional traits (CU) (n = 22) showed deficits relative to low CU ODD boys (n = 25) in emotion perspective-taking and in understanding ambivalent emotions. Low CU ODD boys did not differ from the healthy typically developing boys (n = 12). Impairments in emotion perceptive-taking and understanding mixed emotions in ODD boys are associated with the presence of a high level of CU.


Use of molecular beacons to image effects of titanium surface microstructure on beta1 integrin expression in live osteoblast-like cells.
This study used molecular beacon technology to examine substrate-dependent changes in integrin subunit expression in living cells. Molecular beacons are oligonucleotide probes that can be delivered into live cells to allow for real-time imaging of mRNA. They have a stem-loop hairpin structure with a fluorophore-quencher pair, which opens when bound to the target mRNA sequence, resulting in a fluorescent signal upon excitation. A novel molecular beacon that is specific to the beta1 integrin subunit mRNA was developed and used to image osteoblast-like MG63 cells in vitro on both glass and titanium surfaces of varying roughness. Specificity was verified by comparing the molecular beacon signal intensities to real-time PCR results in both wild-type cells and cells with shRNA knockdown of beta1 integrin mRNA. The molecular beacon was able to detect changes due to both surface microtopography and silencing of the mRNA target. The results showed that effects of the substrate on beta1 mRNA noted previously in confluent cultures were evident in pre-confluent cells as well, supporting the hypothesis that beta1 integrin pairs are important in proliferation as well as differentiation of osteoblasts. This technique overcomes the limitations of traditional gene assays (PCR, immunofluorescence) by allowing for the real-time measurement and tracking of specific mRNAs in individual live cells prior to confluence.


BRD4 Inhibition Is Synthetic Lethal with PARP Inhibitors through the Induction of Homologous Recombination Deficiency.
Poly(ADP-ribose) polymerase inhibitors (PARPi) are selectively active in cells with homologous recombination (HR) deficiency (HRD) caused by mutations in BRCA1, BRCA2, and other pathway members. We sought small molecules that induce HRD in HR-competent cells to induce synthetic lethality with PARPi and extend the utility of PARPi. We demonstrated that inhibition of bromodomain containing 4 (BRD4) induced HRD and sensitized cells across multiple tumor lineages to PARPi regardless of BRCA1/2, TP53, RAS, or BRAF mutation status through depletion of the DNA double-stand break resection protein CtIP (C-terminal binding protein interacting protein). Importantly, BRD4 inhibitor (BRD4i) treatment reversed multiple mechanisms of resistance to PARPi. Furthermore, PARPi and BRD4i are synergistic in multiple in vivo models.


Nanomaterials as a Sustainable Choice for Treating Wastewater: A Review.
The removal of dyes from textile effluents utilizing advanced wastewater treatment methods with high efficiency and low cost has received substantial attention due to the rise in pollutants in water. The purpose of this work is to give a comprehensive analysis of the different treatments for removing chemical dyes from textile effluents. The capability and potential of conventional treatments for the degradation of dyeing compounds in aqueous media, as well as the influence of multiple parameters, such as the pH solution, initial dye concentration, and adsorbent dose, are presented in this study. This study is an overview of the scientific research literature on this topic, including nanoreductive and nanophotocatalyst processes, as well as nanoadsorbents and nanomembranes. For the purpose of treating sewage, the special properties of nanoparticles are currently being carefully researched. The ability of nanomaterials to remove organic matter, fungus, and viruses from wastewater is another benefit. Nanomaterials are employed in advanced oxidation techniques to clean wastewater. Additionally, because of their small dimensions, nanoparticles have a wide effective area of contact. Due to this, nanoparticles' adsorption and reactivity are powerful. The improvement of nanomaterial technology will be beneficial for the treatment of wastewater. This report also offers a thorough review of the distinctive properties of nanomaterials used in wastewater treatment, as well as their appropriate application and future possibilities. Since only a few types of nanomaterials have been produced, it is also important to focus on their technological feasibility in addition to their economic feasibility. According to this study, nanoparticles (NPs) have a significant adsorption area, efficient chemical reactions, and electrical conductivity that help treat wastewater effectively.


Radiological and Clinical Outcome After Reversed L-Shaped Osteotomy: A Large Retrospective Swiss Cohort Study.
The objective was to report radiological and clinical outcomes after reversed L-shaped osteotomy (ReveL) for hallux valgus (HV). A retrospective cohort study was performed between January 2004 and December 2013. The primary outcome was radiological recurrence of HV (HV angle [HVA] >15°). There were various exposure and secondary outcome variables. The results showed a median follow-up of 12.0 months (N = 827). Radiological recurrence, limited patient satisfaction, complication, revision surgery, and elective hardware removal were found in 25.0%, 15.3%, 4.6%, 2.5%, and 26.7%. Median pre- to postoperative changes were highest for HVA (delta = -16.7°). Recurrence was more likely in cases with preoperative HVA ≥40° (adjusted odds ratio [ORadjusted]) 3.63, p < .001). Revisions were more likely with concomitant diseases and bilateral surgery (ORadjusted 12.53, p = .010; ORadjusted 3.35, p = .030). Hardware removal was less likely in patients ≥50 years (ORadjusted 0.67, p = .014). In conclusion, ReveL was a good surgical option for HV because of the relatively low rates of unfavorable outcomes.


Computed tomography of chiasmal optic neuritis.
Three cases of chiasmal optic neuritis are described with bitemporal visual field loss and enlargement of the optic chiasm as demonstrated by computed tomography (CT). Exploratory craniotomies were performed in two of the patients, but no tumor was found. After corticosteroid treatment, all three patients experienced virtually complete restoration of normal vision. Although the CT presentation in these patients was indistinguishable from that of a chiasmal glioma, the clinical presentation of acute visual loss in an older child or adult should suggest chiasmal optic neuritis. A period of observation, with or without a trial of corticosteroids, might permit the latter diagnosis and obviate exploratory craniotomy.


Thermafil obturation: a literature review.
A review of the literature pertaining to the use of Thermafil Endodontic Obturators is presented in this article. Addressed are such concerns as apical microleakage, biocompatibility, coronal leakage, post space considerations, retreatment issues, surgical factors, and clinical evaluations of Thermafil; included are the authors' conclusions concerning its clinical use.


Development and validation of a WHOQOL-BREF Taiwanese audio player-assisted interview version for the elderly who use a spoken dialect.
A quality of life questionnaire is rarely adapted to an interview mode for people who mainly use spoken language in daily life. In Taiwan, the WHOQOL-BREF (Mandarin Chinese version) has been developed, as a self-administered questionnaire, but it cannot be applied to the majority of the elderly in Taiwan, who speak only Taiwanese (a dialect). This study adopted the audio player-assisted interview mode to develop a Taiwanese version of the WHOQOL-BREF specifically for Taiwanese-speaking elderly people, and followed with examinations of the reliability and validity of this version. Initially, the WHOQOL-BREF (English version) was translated into colloquial Taiwanese, and field tests confirmed the equivalence and appropriateness of the translation. A total of 228 Taiwanese-speaking elderly people were assessed using the Taiwanese interview version, of which 144 subjects were re-assessed two weeks later. Interviewers assessed each subject aided by an audio player on which all the translated WHOQOL-BREF contents were recorded. The Taiwanese interview version of the WHOQOL-BREF, except for the item related to dependence on medication, showed acceptable reliability (internal consistency, corrected item-domain correlation, and test-retest reliability) and validity (criterion-related, convergent, and discriminant validity). Confirmatory factor analyses supported the four-factor model of the Taiwanese interview version, providing evidence for construct validity. The results suggest that the Taiwanese audio player-assisted interview version of the WHOQOL-BREF was reliable and valid in assessing quality of life of elderly Taiwanese.


Dabrafenib and trametinib treatment-associated fevers in metastatic melanoma causing extreme elevation in procalcitonin in the absence of infection.
Febrile illnesses are common in the management of metastatic solid organ malignancies. Traditionally they occur in the setting of immunosuppression and neutropenia owing to cytotoxic therapy necessitating consideration of systemic infections. Systemic markers of inflammation, such as C-reactive protein and procalcitonin (PCT), may be used to assist in determining the aetiology of a fever in such patients. Newer anticancer therapies may cause significant noninfectious fevers and may result in a rise in inflammatory markers, despite the absence of an infection. We present a case of a critically unwell febrile patient being treated with dabrafenib and trametinib for advanced melanoma. The patient had an extreme elevation in PCT in the absence of infection. We discuss the presentation of fevers related to dabrafenib and trametinib therapy in the management of advanced melanoma, and the utility of PCT in the management of fevers in advanced solid organ malignancies.


Defect formation and migration in Nasicon Li1+xAlxTi2-x(PO4)3.
NASICON-structured materials of the composition Li1+xAlxTi2-x(PO4)3 are regarded as solid electrolytes with high Li-ion conductivity applicable in all solid-state batteries. In this study we investigate the migration paths of constituting ions and monovalent charge carriers including K+, Na+, and, H+. The results proof that Li is the most mobile species in the investigated composition and that the formation of intrinsic defects is unlikely. In addition, we find surprisingly low migration energy for oxygen vacancies in the structure of the dedicated Li-ion conductor.


Using ecological momentary assessment to determine media use by individuals with and without major depressive disorder.
To use ecological momentary assessment techniques to measure the association of major depressive disorder (MDD) with media use. Data were collected using an ecological momentary assessment protocol with cellular telephone-based brief interviews. Participants received as many as 60 telephone calls from a trained staff member during 5 extended weekends in an 8-week period. One hundred six adolescent participants who were part of a larger neurobehavioral study of depression in Pittsburgh from January 1, 2003, through December 31, 2008. At each call, participants were asked whether they were using the following 5 types of media: television or movies, music, video games, Internet, and print media, such as magazines, newspapers, and books. We developed multivariable models to determine the independent association of each type of media use with MDD, controlling for sociodemographic variables. Of the 106 participants, 46 were diagnosed as having MDD. In multivariable models controlling for age, sex, and race, each increasing quartile of audio use was associated with an 80% increase in the odds of having MDD (odds ratio, 1.8; 95% confidence interval, 1.1-2.8; P = .01 for trend). Conversely, each increasing quartile of print media use was associated with a 50% decrease in the odds of having MDD (odds ratio, 0.5; 95% confidence interval, 0.3-0.9; P = .009 for trend). Major depressive disorder is positively associated with popular music exposure and negatively associated with reading print media such as books. Further research elucidating the directionality and strength of these relationships may help advance understanding of the relationships between media use and MDD.


Electrochemical Removal of Organic and Inorganic Pollutants Using Robust Laser-Induced Graphene Membranes.
The removal of emerging environmental pollutants in water and wastewater is essential for high drinking water quality or for discharge to the environment. Electrochemical treatment is a promising technology shown to degrade undesirable organic compounds or metals via oxidation and reduction, and carbon-based electrodes have been reported. Here, we fabricated a robust, porous laser-induced graphene (LIG) electrode on a commercial water treatment membrane using the multilasing technique and demonstrated the electrochemical removal of iohexol, an iodine contrast compound, and chromium(VI), a highly toxic heavy metal ion. Multiple lasing resulted in a more ordered graphitic lattice, a more physically robust carbon layer, and a 3-4-fold higher electrical conductivity. These properties ultimately led to a more efficient electrochemical process, and the optimized LIG electrodes showed a higher hydrogen peroxide (H2O2) generation. At 3 V, 90% of Cr(VI) was removed after 6 h and reached >95% removal after 8 h at pH 2. Cr(VI) was mainly reduced to Cr(III), with small amounts of Cr(I) and Cr(0), which were partially deposited on the electrode membrane surface, confirmed with X-ray photoelectron spectroscopy and energy-dispersive X-ray spectroscopy analysis. Under the same conditions, 50% of iohexol was degraded after 6 h and the transformation products (TPs) were identified using ultra-performance liquid chromatography coupled with mass spectroscopy. A total of seven main intermediates were identified including deiodinated TPs (m/z = 695, 570, and 443), probably occurring via three transformation pathways including oxidative deiodination, amide hydrolysis, and deacetylation. The electrical energy costs calculated for the removal of 2 mg L-1 Cr(VI) was ∼$0.08/m3 in this system. Taken together, the porous LIG electrodes might be utilized for electrochemical removal of emerging contaminants in multiple applications because they can be rapidly formed on flexible polymer substrates at low cost.


Generation of random numbers by measuring phase fluctuations from a laser diode with a silicon-on-insulator chip.
Random numbers are a fundamental resource in science and technology. Among the different approaches to generating them, random numbers created by exploiting the laws of quantum mechanics have proven to be reliable and can be produced at enough rates for their practical use. While these demonstrations have shown very good performance, most of the implementations using free-space and fibre optics suffer from limitations due to their size, which strongly limits their practical use. Here we report a quantum random number generator based on phase fluctuations from a diode laser, where the other required optical components are integrated on a mm-scale monolithic silicon-on-insulator chip. The post-processing reported in this experiment is performed via software. However, our physical device shows the potential of operation at generation rates in the Gbps regime. Considering the device's size, its simple, robust and low power operation, and the rapid industrial uptake of silicon photonics, we foresee the widespread integration of the reported design in more complex systems.


Real-time cylindrical curvilinear 3-D ultrasound imaging.
In patients who are obese or exhibit signs of pulmonary disease, standard transthoracic scanning may yield poor quality cardiac images. For these conditions, two-dimensional transesophageal echocardiography (TEE) is established as an essential diagnostic tool. Current techniques in transesophageal scanning, though, are limited by incomplete visualization of cardiac structures in close proximity to the transducer. Thus, we propose a 2D curvilinear array for 3D transesophageal echocardiography in order to widen the field of view and increase visualization close to the transducer face. In this project, a 440 channel 5 MHz two-dimensional array with a 12.6 mm aperture diameter on a flexible interconnect circuit has been molded to a 4 mm radius of curvature. A 75% element yield was achieved during fabrication and an average -6dB bandwidth of 30% was observed in pulse-echo tests. Using this transducer in conjunction with modifications to the beam former delay software and scan converter display software of the our 3D scanner, we obtained cylindrical real-time curvilinear volumetric scans of tissue phantoms, including a field of view of greater than 120 degrees in the curved, azimuth direction and 65 degrees phased array sector scans in the elevation direction. These images were achieved using a stepped subaperture across the cylindrical curvilinear direction of the transducer face and phased array sector scanning in the noncurved plane. In addition, real-time volume rendered images of a tissue mimicking phantom with holes ranging from 1 cm to less than 4 mm have been obtained. 3D color flow Doppler results have also been acquired. This configuration can theoretically achieve volumes displaying 180 degrees by 120 degrees. The transducer is also capable of obtaining images through a curvilinear stepped subaperture in azimuth in conjunction with a rectilinear stepped subaperture in elevation, further increasing the field of view close to the transducer face. Future work includes development of an array for adapting these modifications to a 6 mm diameter endoscope probe.


Pseudomonas aeruginosa serious infections: mono or combination antimicrobial therapy?
P. aeruginosa is a serious cause of infection with reported rates of mortality being up to 61%. Several studies evidenced a correlation between hospital mortality due to P. aeruginosa bloodstream infections and an inappropriate antimicrobial treatment. Increasing resistance in P. aeruginosa isolates complicates the selection of adequate empirical therapy in severe infections and P. aeruginosa is often indistinguishable from other gram-negative bacterial infections. For these reasons, present guidelines for the treatment of suspected P. aeruginosa bacteraemia recommend the rapid introduction of empirical antimicrobial therapy that includes at least one antipseudomonal agent until having microbiological results. Current consensus favours the use of empirical combination, balancing the potential for greater toxicity against the lower emergence of antimicrobial resistance and the greater killing that might be achieved by combination therapies acting synergistically. Advantages and disadvantages of combination therapy towards monotherapy for P. aeruginosa severe infections, current antibiotics used for P. aeruginosa severe infections and main studies published on this issue are reviewed.


Advances in the Discovery of Novel Inhaled PI3Kδ Inhibitors for the Treatment of Asthma.
Bronchial asthma is the most common chronic respiratory illness, the incidence of which continues to increase annually. Currently, effective treatments for CS-resistant asthma and severe asthma are still lacking, and new therapeutic regimens are urgently required. PI3Kδ is a key enzyme in hematopoietic cells and represents a major target for oncology and inflammatory disease (particularly respiratory disease, asthma and COPD). In the case of respiratory disease, the ability to inhibit PI3Kδ in the lungs shows a higher safety and therapeutic index relative to systemic inhibition. In recent years, paradigm shifts have occurred in inhalation therapeutics for systemic and topical drug delivery due to the favorable properties of lungs, including their large surface area and high permeability. Pulmonary drug delivery possesses many advantages, including a non-invasive route of administration, low metabolic activity, a controlled environment for systemic absorption and the ability to avoid first bypassing metabolism. In this review, we focus on the discovery and development of inhaled drugs targeting PI3Kδ for asthma by focusing on their activity and selectivity, in addition to their potential in drug design strategies using inhaled administration.


Long-term follow-up and outcomes of discrete subaortic stenosis resection in children.
Studies of long-term outcomes of discrete subaortic stenosis (DSS) are rare. Therefore, we reviewed the long-term outcomes of subaortic membrane resection in children with isolated DSS over 16 years from a single institution. We retrospectively reviewed the records of patients (n = 27) who underwent resection of DSS between 2000 and 2017. Patients with major concomitant intracardiac anomalies were excluded. Indications for surgery were mean left ventricular outflow tract (LVOT), Doppler gradient >30 mmHg, and/or progressive aortic insufficiency. The mean age at diagnosis was 3.77 ± 3.49 years (range, 0.25-13 years) and the mean age at surgery was 6.36 ± 3.69 years (range, 1-13 years). All patients underwent resection of subaortic membrane. The mean LVOT Doppler gradient decreased from 40.52 ± 11.41 mmHg preoperatively to 8.48 ± 5.06 mmHg postoperatively (P < 0.001). The peak instantaneous LVOT Doppler gradient decreased from 75.41 ± 15.22 mmHg preoperatively to 18.11 ± 11.44 mmHg postoperatively (P < 0.001). At the latest follow-up, the peak gradient was 17.63 ± 8.93 mmHg. The mean follow-up was 7.47 ± 3.53 years (median 6.33 years; range 2.67-16 years). There was no operative mortality or late mortality. Recurrence of subaortic membrane occurred in 7 (25.92%, 7/27) patients who underwent primary DSS operation. Four (14.81%, 4/27) patients required reoperation for DSS recurrence at a median time of 4.8 years (3.1-9.1 years) after the initial repair. Risk factors for reoperation were age <6 years at initial repair. Eighteen (66.66%, 18/27) patients had AI preoperatively and progression of AI occurred in 70.37% (19/27). This included 4 (22.22%, 4/18) patients who had worsening of their preoperative AI. Short valve-to-membrane distance was found to be prognostically unfavorable. One (3.7%, 1/27) patient had an iatrogenic ventricular septal defect, and 2 (7.4%, 2/27) patients had complete AV block following membrane resection. Resection of subaortic membrane in children is associated with low mortality. Higher LVOT gradient, younger age at initial repair, and shorter valve-to-membrane distance were found to be associated with adverse outcome. Recurrence and reoperation rates are high, and progression of aortic insufficiency following subaortic membrane resection is common. Therefore, these patients warrant close follow-up into adult life.


Similar odorants elicit different behavioral and physiological responses, some supersustained.
An intriguing question in the field of olfaction is how animals distinguish among structurally similar odorants. We systematically analyzed olfactory responses elicited by a panel of 25 pyrazines. We found that structurally similar pyrazines elicit a wide range of behavioral responses from Drosophila larvae. Each pyrazine was tested against all functional receptors of the larval Odor receptor (Or) repertoire, yielding 525 odorant-receptor combinations. Different pyrazines vary markedly in the responses they elicit from the Or repertoire, with most strong responses deriving from two receptors, Or33b and Or59a. Surprisingly, 2-ethylpyrazine and 2-methylpyrazine, which elicit strikingly similar physiological responses across the receptor repertoire, elicit dramatically different behavioral responses. A small fraction of odorant-receptor combinations elicit remarkably long responses. These responses, which we term "supersustained" responses, are receptor specific and odorant specific, and can last for minutes. Such supersustained responses may prevent olfactory neurons from reporting contemporaneous information about the local odor environment. Odors that elicit such responses could provide a novel means of controlling insect pests and vectors of human disease by impairing the location of human hosts, food sources, and mates.


Subgroups in Late Adulthood Are Associated With Cognition and Wellbeing Later in Life.
Objectives: In this study, we aim to discover whether there are valid subgroups in aging that are defined by modifiable factors and are determinant of clinically relevant outcomes regarding healthy aging. Method: Data from interviews were collected in the Longitudinal Aging Study Amsterdam at two measurement occasions with a 3-year interval. Input for the analyses were seven well-known vulnerability and protective factors of healthy aging. By means of community detection, we tested whether we could distinguish subgroups in a sample of 1478 participants (T1-sample, aged 61-101 years). We tested both the external validity (T1) and predictive validity (T2) for wellbeing and subjective cognitive decline. Moreover, replicability and long-term stability were determined in 1186 participants (T2-sample, aged 61-101 years). Results: Three similar subgroups were identified at T1 and T2. Subgroup A was characterized by high levels of education with personal vulnerabilities, subgroup B by being physically active with low support and low levels of education, and subgroup C by high levels of support with low levels of education. Subgroup C showed the lowest wellbeing and memory profile, both at T1 and T2. On most measures of wellbeing and memory, subgroups A and B did not differ from each other. At T2, the same number of subgroups was identified and subgroup profiles at T1 and T2 were practically identical. Per T1 subgroup 47-62% retained their membership at T2. Discussion: We identified valid subgroups that replicate over time and differ on external variables at current and later measurement occasions. Individual change in subgroup membership over time shows that transitions to subgroups with better outcomes are possible.


Separating phonological and semantic processing in auditory sentence processing: a high-resolution event-related brain potential study.
Phonological and semantic processing was studied using high-resolution event-related brain potentials (ERPs) during a sentence-matching task to investigate the spatial distribution of the phonological mismatch negativity (PMN) and the N400 response. It was hypothesized that the two components were spatially separable and that the activity matched prior localization knowledge. Participants examined visual-auditory sentence pairs that related within a semantic hierarchy (e.g., visual: "The man is teaching in the classroom"; Auditory: "The man is in the em leader school/barn"). Semantic congruency was varied for the final words of the spoken sentences. Incongruent words mismatched expectation in terms of both the initial phonological features (unexpected sound) and semantic features (unexpected meaning). In addition, the category-exemplar probability of the final words was either high or low, with low probability words being more difficult to anticipate. Low probability words were predicted to selectively affect PMN activity. We found that incongruent words elicited a PMN (287 msec) and a N400 (424 msec), for both the high and low probability words. As expected, low probability congruent words elicited a small PMN but no N400. In contrast, high probability congruent words elicited neither a detectible PMN nor a N400. The primary PMN sources were in left inferior frontal and inferior parietal lobes. The primary N400 source activation occurred along the left perisylvian cortex, consistent with prior N400 source localization work. From these results, it was concluded that the PMN and N400 were localized to separate cortical language (and memory) regions and had different source activation patterns.


Pain modulatory network is influenced by sex and age in a healthy state and during osteoarthritis progression in rats.
Old age and female sex are risk factors for the development of osteoarthritis (OA) and chronic pain. We investigated the effects of sex and age on pain modulatory networks in a healthy state and during OA progression. We used functional MRI to determine the effects of sex and age on periaqueductal gray functional connectivity (PAG FC) in a healthy state (pre-OA) and during the early and late phases of monosodium iodoacetate-induced OA in rats. We then examined how sex and age affect longitudinal changes in PAG FC in OA. In a healthy state, females exhibited more widespread PAG FC than males, and this effect was exaggerated with aging. Young males had moderate PAG FC changes during the early phase but recruited additional brain regions, including the rostral anterior cingulate cortex (ACC), during the late phase. Young females exhibited widespread PAG FC in the early phase, which includes connections to insula, caudal ACC, and nucleus accumbens (NAc). Older groups had strong PAG FC with fewer regions in the early phase, but they recruited additional brain regions, including NAc, in the late phase. Overall, our findings show that PAG FC is modulated by sex and age in a healthy state. A widespread PAG network in the early phase of OA pain may contribute to the transition from acute to chronic OA pain and the increased risk of developing chronic pain for females. Enhanced PAG FC with the reward system may represent a potential mechanism underlying chronic OA pain in elderly patients.


Identification of novel androgen-regulated pathways and mRNA isoforms through genome-wide exon-specific profiling of the LNCaP transcriptome.
Androgens drive the onset and progression of prostate cancer (PCa) by modulating androgen receptor (AR) transcriptional activity. Although several microarray-based studies have identified androgen-regulated genes, here we identify in-parallel global androgen-dependent changes in both gene and alternative mRNA isoform expression by exon-level analyses of the LNCaP transcriptome. While genome-wide gene expression changes correlated well with previously-published studies, we additionally uncovered a subset of 226 novel androgen-regulated genes. Gene expression pathway analysis of this subset revealed gene clusters associated with, and including the tyrosine kinase LYN, as well as components of the mTOR (mammalian target of rapamycin) pathway, which is commonly dysregulated in cancer. We also identified 1279 putative androgen-regulated alternative events, of which 325 (∼25%) mapped to known alternative splicing events or alternative first/last exons. We selected 30 androgen-dependent alternative events for RT-PCR validation, including mRNAs derived from genes encoding tumour suppressors and cell cycle regulators. Of seven positively-validating events (∼23%), five events involved transcripts derived from alternative promoters of known AR gene targets. In particular, we found a novel androgen-dependent mRNA isoform derived from an alternative internal promoter within the TSC2 tumour suppressor gene, which is predicted to encode a protein lacking an interaction domain required for mTOR inhibition. We confirmed that expression of this alternative TSC2 mRNA isoform was directly regulated by androgens, and chromatin immunoprecipitation indicated recruitment of AR to the alternative promoter region at early timepoints following androgen stimulation, which correlated with expression of alternative transcripts. Together, our data suggest that alternative mRNA isoform expression might mediate the cellular response to androgens, and may have roles in clinical PCa.


PIERS proteinuria: relationship with adverse maternal and perinatal outcome.
To examine the ability of three different proteinuria assessment methods (urinary dipstick, spot urine protein:creatinine ratio [Pr/Cr], and 24-hour urine collection) to predict adverse pregnancy outcomes. We performed a prospective multicentre cohort study, PIERS (Preeclampsia Integrated Estimate of RiSk), in seven academic tertiary maternity centres practising expectant management of preeclampsia remote from term in Canada, New Zealand, and Australia. Eligible women were those admitted with preeclampsia who had at least one antenatal proteinuria assessment by urinary dipstick, spot urine Pr/Cr ratio, and/or 24-hour urine collection. Proteinuria assessment was done either visually at the bedside (by dipstick) or by hospital clinical laboratories for spot urine Pr/Cr and 24-hour urine collection. We calculated receiver operating characteristic area under the curve (95% CI) for each proteinuria method and each of the combined adverse maternal outcomes (within 48 hours) or adverse perinatal outcomes (at any time). Models with AUC ≥ 0.70 were considered of interest. Analyses were run for all women who had each type of proteinuria assessment and for a cohort of women ("ALL measures") who had all three proteinuria assessments. More women were proteinuric by urinary dipstick (≥ 2+, 61.4%) than by spot urine Pr/Cr (≥ 30 g/mol, 50.4%) or 24-hour urine collection (≥ 0.3g/d, 34.7%). Each proteinuria measure evaluated had some discriminative power, and dipstick proteinuria (categorical) performed as well as other methods. No single method was predictive of adverse perinatal outcome. The measured amount of proteinuria should not be used in isolation for decision-making in women with preeclampsia. Dipstick proteinuria performs as well as other methods of assessing proteinuria for prediction of adverse events.


CCL19 and CXCL12 trigger in vitro chemotaxis of human mantle cell lymphoma B cells.
Few data are available in the literature on chemokine receptor expression and migratory capability of mantle cell lymphoma (MCL) B cells. Information on these issues may allow us to identify novel mechanisms of chemokine-driven tumor cell migration. The research was designed to investigate: (a) expression of CCR1 to CCR7 and CXCR1 to CXCR5 chemokine receptors; and (b) chemotaxis to the respective ligands in MCL B cells and in their normal counterparts, i.e., CD5+ B cells. Malignant B cells from MCL patients and normal counterparts displayed similar chemokine receptor profiles. MCL B cells were induced to migrate by CXCL12 and CCL19, whereas normal CD5+ B cells migrated to the former, but not the latter chemokine. Overnight culture of MCL B cells and their normal counterparts with CXCL12 cross-sensitized other chemokine receptors to their ligands in some tumor samples but not in CD5+ B cells. CCR7 and CXCR4 ligands may play a key role in tumor cell migration and spreading in vivo. CXCL12 may additionally contribute by sensitizing MCL B cells to respond to the ligands of other chemokine receptors.


Short term modulation of trunk neuromuscular responses following spinal manipulation: a control group study.
Low back pain (LBP) is one of the most frequent musculoskeletal conditions in industrialized countries and its economic impact is important. Spinal manipulation therapy (SMT) is believed to be a valid approach in the treatment of both acute and chronic LBP. It has also been shown that SMT can modulate the electromyographic (EMG) activity of the paraspinal muscle. The purpose of this study was to investigate, in a group of patients with low back pain, the persistence of changes observed in trunk neuromuscular responses after a spinal manipulation (SMT). Sixty adult participants with LBP performed a block of 5 flexion-extension movements. Participants in the experimental group (n=30) received lumbar SMT whereas participants in the control group (n=30) were positioned similarly for the treatment but did not receive SMT. Blocks of flexion-extension movements were repeated immediately after the manipulation as well as 5 and 30 minutes after SMT (or control position). EMG activity of paraspinal muscles was recorded at L2 and L5 level and kinematic data were collected to evaluate the lumbo-pelvic kinematics. Pain intensity was noted after each block. Normalized EMG, pain intensity and lumbo-pelvic kinematics were compared across experimental conditions. Participants from the control group showed a significant increase in EMG activity during the last block (30 min) of flexion-extension trials in both flexion and full-flexion phases at L2. Increase in VAS scores was also observed in the last 2 blocks (5 min and 30 min) in the control group. No significant group x time interaction was seen at L5. No significant difference was observed in the lumbo-pelvic kinematics. Changes in trunk neuromuscular control following HVLA spinal manipulation may reduce sensitization or muscle fatigue effects related to repetitive movement. Future studies should investigate short term changes in neuromuscular components, tissue properties and clinical outcomes.


An electrophoretic study of endogenous phosphorylation in vitro of the polypeptides of microsomal membrane fractions of mouse liver.
1. The patterns of phosphopolypeptides produced by endogenous phosphorylation in vitro of rough- and smooth-membrane fractions of the microsomal fraction of mouse liver were studied by radioautographic analysis of sodium dodecyl sulphate/polyacrylamide-gel electrophoretograms. 2. A minimum of 17 polypeptides of both rough- and smooth-microsomal-membrane fractions were phosphorylated by using [gamma-(32)P]-ATP as the phosphate donor; only minor differences in phosphorylation pattern between the two membrane fractions were detected. 3. Phosphorylation in vitro by [gamma-(32)P]ATP was markedly stimulated by Mg(2+), but not by cyclic AMP, cyclic GMP or Ca(2+). The phosphorylation of certain polypeptides was preferentially stimulated by Mg(2+). Addition of cyclic AMP resulted in a decrease in the amount of (32)P detected in one polypeptide of mol.wt. approx. 56000, present in both the rough- and smooth-membrane fractions. 4. [gamma-(32)P]GTP was found to be a relatively poor donor of (32)P as compared with [gamma-(32)P]ATP. However, incubation of rough- and smooth-membrane fractions with this compound resulted in the phosphorylation of one polypeptide of mol.wt. approx. 96000 that was scarcely or not at all phosphorylated by [gamma-(32)P]ATP. 5. Under the conditions of incubation used, appreciable incorporation of (32)P from [gamma-(32)P]ATP occurred into products migrating at the front of the electrophoretograms; these products were identified as being principally comprised of 1-phosphatidylinositol 4-phosphate. Incorporation of (32)P into this lipid was also markedly stimulated by Mg(2+). 6. The overall results show that a considerable number of polypeptides of the rough- and smooth-microsomal-membrane fractions of mouse liver may be phosphorylated in vitro and indicate that the enzymes responsible are principally non-cyclic AMP-dependent protein kinases.