Cytomegalovirus disease and infection in lung transplant recipients in the setting of planned indefinite valganciclovir prophylaxis.
The optimal method of both predicting and preventing cytomegalovirus (CMV) disease in lung transplant recipients remains unclear. In particular, the most appropriate duration of CMV prophylaxis post transplant is unresolved. We report herein our experience with a planned indefinite regimen of valganciclovir prophylaxis and monitoring of quantitative CMV load in bronchoalveolar lavage (BAL) fluid. We performed a single-center observational study with both prospective and retrospective components. The included patients (n = 128) received a planned regimen of indefinite valganciclovir prophylaxis post transplant, regardless of donor (D)/recipient (R) CMV serostatus. Real-time polymerase chain reaction assay for detection of CMV in BAL was prospectively performed over a 1-year period. Clinical data were reviewed retrospectively; median follow-up was 24.8 months (range 1-93 months). Sixty-five patients (50.6%) discontinued valganciclovir prophylaxis, either temporarily or permanently, with a primary cause of mild leukopenia. Six cases of CMV disease were identified (4.7%), with no significant difference between those who were on continuous prophylaxis or not (4.6% vs. 4.9%; P = non-significant [ns]). However, those who discontinued prophylaxis showed an increased incidence of laboratory-detected CMV infection (40.7% vs. 12.7%; P = 0.001). High-risk D+/R- patients did not demonstrate a significantly increased incidence of CMV disease (8.1% vs. 3.3% other serotypes; P = ns). Three patients (2.3%) developed valganciclovir-resistant CMV disease. Molecular detection of CMV in BAL fluid was significantly more sensitive than shell vial culture. However, BAL CMV viral load was not predictive of subsequent disease development. Extended valganciclovir prophylaxis for all lung transplant recipients led to a low incidence of CMV disease and resistance. In such low-incidence populations, routine quantitation of CMV in BAL did not confer significant clinical benefit over non-quantitative methods in prediction of CMV disease onset.


Unmetabolized fenofibrate, but not fenofibric acid, activates AMPK and inhibits the expression of phosphoenolpyruvate carboxykinase in hepatocytes.
A lipid-lowering agent, fenofibrate, has been reported to reduce hepatic glucose production and the expression of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme for gluconeogenesis in the liver. However, the precise mechanisms of these effects have remained unclear. Rat hepatoma-derived H4IIE cells and murine myoblast-derived C2C12 cells were incubated with the ester form of fenofibrate and fenofibric acid, a metabolite of fenofibrate ester, and the phosphorylation of AMPK, acetyl-CoA carboxylase (ACC), PEPCK mRNA expression and glucose production were assessed. Incubation of H4IIE hepatoma cells with the ester form of fenofibrate increased the phosphorylation of AMPK and acetyl-CoA carboxylase (ACC) and decreased PEPCK mRNA expression and glucose production. Fenofibrate-induced reductions in PEPCK expression and glucose production were abrogated by compound C, a specific AMPK inhibitor. Fenofibric acid, a metabolite of fenofibrate ester, had no effects on AMPK phosphorylation, PEPCK gene expression, or glucose production in H4IIE cells. Fenofibrate-treated mice exhibited increases in AMPK phosphorylation and a decrease in PEPCK expression in the liver but not in skeletal muscles, suggesting that unmetabolized fenofibrate accumulated and affected AMPK only in the liver. These results demonstrate that fenofibrate inhibits PEPCK gene expression and hepatic glucose production in the liver via AMPK activation, even though the metabolite loses its effects on AMPK and does not work in vivo in myocytes. This novel feature of fenofibrate may provide additional benefit for the treatment of patients with disorders of both lipid and glucose metabolism.


An implementation evaluation of the community engagement and planning intervention in the CPIC Depression Care Improvement Trial.
The goal of this paper is to document and evaluate the process of implementing an evidence-based depression intervention in community settings through the use of community-academic partnered approaches. We discuss how and to what extent the goals of community engagement and collaborative planning were achieved in the intervention arm of the Community Partners in Care study that aimed to adapt evidence-based depression care toolkits for diverse agencies in Hollywood and South Los Angeles. We find that partnered research strategies have a potential to effectively engage community members around depression and involve them in intervention planning activities. Our results suggest that successful collaboration among diverse agencies requires that they understand what is expected of them, are comfortable with the role they choose to perform, and have organizational support to contribute to the project. To facilitate the development of collaborative relationships, time and effort should be devoted to explaining how collaboration among diverse agencies may take place.


Electron spin resonance in ATP and RNA.
Cu(++), Mn(++), and Fe(+++) account for the electron spin resonances observed in certain samples of ATP and RNA. The copper ion seems more loosely bound to these substances than either iron or manganese. A striking similarity is observed between the manganese spectra in manganese RNA, ATP, and ADP suggesting that the binding sites are similar in the three compounds. The similarity of the e.s.r. spectra of iron ATP and of iron and manganese RNA, except for hyperfine spectrum (hfs) in the latter, suggests that the two ions bind similarly in the two compounds. A detailed interpretation of the spectra is lacking however and these conclusions can only be tentative. When manganese TPP and ATP are heated or pH changed the e.s.r. alters indicating a change in the environment of the ion. The sharp 6 line manganese spectrum in both TPP and ATP at pH 1 suggests an almost "free" ion at this pH in the sense of an almost isotropic average environment.


An eye movement study of insight problem solving.
The representational change theory of insight claims that insight problems cause impasses because they mislead problem solvers into constructing inappropriate initial representations. Insight is attained when the initial representation is changed. In the present study (N = 24), we tested three specific implications of these hypotheses against eye movements recorded while participants solved matchstick arithmetic problems. The results were consistent with the predictions, providing converging evidence with prior findings using solution rates and solution times. Alternative theories of insight can explain individual findings, but only the representational change theory accounts for both the performance data and the eye movement data. The present study also suggests that eye movement recordings provide an important new window into processes of insight problem solving.


Diagnostic Value of MicroRNAs for Urologic Cancers: A Systematic Review and Meta-Analysis.
MicroRNAs (miRNAs), particularly those extracted from the blood or tissues, have become the focus of urologic cancers research. However, the literature reviews on the accuracy of miRNA detection in urologic cancers have been inconsistent, leading us to perform this meta-analysis. Eligible studies were searched in PubMed and other databases. To calculate the pooled detection accuracy estimates, we used a bivariate random-effects meta-analysis model. According to the exclusion and inclusion criteria, 41 studies were included. Overall, the results showed sensitivity of 0.77 (95% CI: 0.74-0.80) and specificity of 0.76 (95% CI: 0.72-0.79), with an area under the SROC curve (AUC) of 0.83 (95% CI: 0.80-0.86). In addition, further subgroup analyses were also conducted. Firstly, the multiple miRNAs subgroup has significantly better diagnostic specificity than single miRNA subgroup among all these cancer types, while only bladder cancer (BC) and prostate cancer (PC) group with significantly greater diagnostic sensitivity with their multiple miRNA detection. Secondly, none of these cancer types showed significant differences on diagnostic sensitivity and specificity in their specimen and sample size subgroups. Thirdly, the diagnostic sensitivity between Asian (0.791, 95% CI: 0.748-0.827) and Caucasian (0.713, 95% CI: 0.666-0.756) in BC type was shown significant different with the P-value of 0.011. The results of our study suggested that miRNAs, particularly the multiple miRNAs, may play an important role in diagnosis and monitoring of the urologic cancers as superior biomarkers.


Robotic tubal anastomosis: technical aspects.
To describe the surgical technique of robotic tubal anastomosis. Retrospective chart and video review of the instrumentation and methodology used for robotically assisted tubal anastomosis. All tubal anastomoses were performed with the use of 3 or 4 robotic arms, 3 or 4 robotic instruments, and 1 assistant trocar. Robotic technology facilitates the performance of robotic tubal anastomosis.


The impact of cryosolution thermal contraction on proteins and protein crystals: volumes, conformation and order.
Cryocooling of macromolecular crystals is commonly employed to limit radiation damage during X-ray diffraction data collection. However, cooling itself affects macromolecular conformation and often damages crystals via poorly understood processes. Here, the effects of cryosolution thermal contraction on macromolecular conformation and crystal order in crystals ranging from 32 to 67% solvent content are systematically investigated. It is found that the solution thermal contraction affects macromolecule configurations and volumes, unit-cell volumes, crystal packing and crystal order. The effects occur through not only thermal contraction, but also pressure caused by the mismatched contraction of cryosolvent and pores. Higher solvent-content crystals are more affected. In some cases the solvent contraction can be adjusted to reduce mosaicity and increase the strength of diffraction. Ice formation in some crystals is found to cause damage via a reduction in unit-cell volume, which is interpreted through solvent transport out of unit cells during cooling. The results point to more deductive approaches to cryoprotection optimization by adjusting the cryosolution composition to reduce thermal contraction-induced stresses in the crystal with cooling.


Metformin confers longitudinal cardiac protection by preserving mitochondrial homeostasis following myocardial ischemia/reperfusion injury.
Myocardial ischemia-reperfusion (I/R) injury is associated with systemic oxidative stress, cardiac mitochondrial homeostasis, and cardiomyocyte apoptosis. Metformin has been recognized to attenuate cardiomyocyte apoptosis. However, the longitudinal effects and pathomechanism of metformin on the regulation of myocardial mitohormesis following I/R treatment remain unclear. This study aimed to investigate the longitudinal effects and mechanism of metformin in regulating cardiac mitochondrial homeostasis by serial imaging with the 18-kDa translocator protein (TSPO)-targeted positron emission tomography (PET) tracer 18F-FDPA. Myocardial I/R injury was established in Sprague-Dawley rats, which were treated with or without metformin (150 mg/kg per day). Serial gated 18F-FDG and 18F-FDPA PET imaging were performed at 1, 4, and 8 weeks after surgery, followed by analysis of ventricular remodelling and cardiac mitochondrial homeostasis. The correlation between Hsp60 and 18F-FDPA uptake was analyzed. After PET imaging, the activity of antioxidant enzymes, immunostaining, and western blot analysis were performed to analyze the spatio-temporal effects and pathomechanism of metformin for cardiac protection after myocardial I/R injury. Oxidative stress and apoptosis increased 1 week after myocardial I/R injury (before significant progression of ventricular remodelling). TSPO expression was correlated with Hsp60 expression and was co-localized with inflammatory CD68+ macrophages in the infarct area, and TSPO uptake was associated with an upregulation of AMPK-p/AMPK and a downregulation of Bcl-2/Bax. However, these effects were reversed with metformin treatment. Eight weeks after myocardial I/R injury (representing the advanced stage of heart failure), 18F-FDPA uptake in myocardial cells in the distal non-infarct area increased without CD68+ expression, whereas the activity decreased with metformin treatment. Taken together, these results show that a prolonged metformin treatment has pleiotropic protective effects against myocardial I/R injury associated with a regional and temporal dynamic balance between mitochondrial homeostasis and cardiac outcome, which were assessed by TSPO-targeted imaging during cardiac remodelling.


Mechanisms of Vasorelaxation Induced by Hexahydrocurcuminin Isolated Rat Thoracic Aorta.
This study was designed to examine the vasorelaxant effects of hexahydrocurcumin (HHC), one of the major natural metabolites of curcumin from Curcuma longa, on rat isolated aortic rings, and the underlying mechanisms. Isometric tension of the aortic rings was recorded using organ bath system. HHC (1 nM to 1 mM) relaxed the endothelium-intact aortic rings pre-contracted with PE and KCl in a concentration-dependent manner. Removal of the endothelium did not alter the effect of HHC-induced relaxation. In Ca(2+)-free Krebs solution, HHC significantly inhibited the CaCl2-induced contraction in high K(+) depolarized rings and suppressed the transient contraction induced by PE and caffeine in a concentration-dependent manner. HHC was also observed to relax phobal-12-myristate-13-acetate (PMA), an activator of protein kinase C (PKC), precontracted aortic rings in a concentration-dependent manner with EC50 values equivalent to 93.36 ± 1.03 μM. In addition, pre-incubation with propranolol (a β-adrenergic receptor blocker) significantly attenuated the HHC-induced vasorelaxation. These results suggest that the vasorelaxant effect of HHC is mediated by the endothelium-independent pathway, probably because of the inhibition of extracellular Ca(2+) influx through voltage-operated Ca(2+) channels and receptor-operated Ca(2+) channels, the inhibition of Ca(2+) mobilization from intracellular stores, as well as inhibition of PKC-mediated Ca(2+)-independent contraction. Moreover, HHC produces vasorelaxant effects probably by stimulating the β-adrenergic receptor.


Secondary reconstruction of the lateral ligaments of the ankle by the Chrisman-Snook technique.
Nineteen patients suffering from chronic post-traumatic instability at the ankle were treated surgically by the Chrisman-Snook technique. Mean follow-up time was 38 months. Each patient reported an initial major inversion trauma, followed by repetitive sprains resulting in apprehension, giving way, pain, and swelling at the ankle. Before surgery all patients engaged in sports were obliged to modify or even stop their activities for reasons related to their ankle problem. After reconstruction instability was abolished in all cases and an excellent or good result was obtained in 18. The three top sportsmen included in our series were all able to return to their pre-injury level of sport without any restrictions. A careful dissection of the branches of the sural nerve is considered to be essential, since postoperative paresthesia or numbness over the sural nerve region was found in six cases. This study confirms the efficacy of the Chrisman-Snook technique in the treatment of ankle instability, even in cases of gross ankle laxity or with patients engaged in very demanding sports.


Magnolol upregulates CHRM1 to attenuate Amyloid-β-triggered neuronal injury through regulating the cAMP/PKA/CREB pathway.
Alzheimer's disease (AD) is a common neurodegenerative disease characterized by neuronal degeneration and hyperphosphorylated Tau. Magnolol is an active component isolated from Magnolia officinalis with potential neuroprotection activity. However, the function and mechanism of magnolol in AD progression is largely uncertain. In present study, the biomarkers related to AD and magnolol were predicted by bioinformatics analyses. The key biomarker levels were predicted by GSE5281 and GSE36980 using AlzData. Cell viability was detected by CCK-8 assay. mRNA and protein levels were examined by qRT-PCR and western blotting assays. Cell apoptosis was investigated by caspase-3 activity and flow cytometry analyses. The cAMP/PKA/CREB signaling was evaluated by ELISA and western blotting analyses. The results showed that CHRM1 was a key biomarker for magnolol against AD progression. Magnolol attenuated Aβ-induced viability inhibition, Tau hyperphosphorylation and apoptosis in SH-SY5Y cells by upregulating CHRM1. In addition, the cAMP signaling might be a potential pathway of CHRM1 in AD. Magnolol contributed to activation of the cAMP/PKA/CREB pathway through enhancing CHRM1 level. Inactivation of the cAMP/PKA/CREB signaling reversed the suppressive effect of magnolol on Tau hyperphosphorylation and apoptosis in Aβ-treated SH-SY5Y cells. As a conclusion, magnolol mitigated Aβ-induced Tau hyperphosphorylation and neuron apoptosis by upregulating CHRM1 and activating the cAMP/PKA/CREB pathway.


The effect of unilateral papillectomy on the juxtaglomerular granulated cells.
The changes of the juxtaglomerular index values in the superficial, deep and whole cortical areas of rat kidneys after unilateral papillectomy were analyzed. On the papillectomized side the JGI values showed an increase in the deep, and a decrease in the superficial cortical zone. The changes proved to be significant during the whole experimental period in the juxtamedullary index and between the 4--14 postoperative days in the superficial index values. The tendency of the changes of JGI in the contralateral kidneys was similar to those of the papillectomized kidneys, but significant differences were registered less constantly. These changes are probably the consequences of haemodynamic factors. Papillectomy is considered an incompetent model for studying the regulatory function of macula densa in the synthesis of renin.


Antigenic relationships of fractionated western diamondback rattlesnake (Crotalus atrox) hemorrhagic toxins and other rattlesnake venoms as indicated by monoclonal antibodies.
Seven hemorrhagic factors have been isolated from Crotalus atrox venom, but their antigenic relationships have not been well studied. In this study, two different monoclonal antibodies, C. atrox peak 8 (CA-P-8) and C. atrox subclone 5 (CA-5+), were produced against two C. atrox venom hemorrhagic fractions and used in an ELISA (enzyme-linked immunosorbent assay) to determine if the hemorrhagic factors in C. atrox venom are antigenically related. The same ELISA test was used to determine cross-reactivity of seven other crude Crotalidae venoms. The two monoclonal antibodies were tested for their ability to neutralize each hemorrhagic HPLC fraction separated from C. atrox venom. C. atrox venom was fractionated into 22 fractions using HPLC analytical DEAE ion exchange. Fractions 4-17 were hemorrhagic. The CA-P-8 monoclonal antibody reacted strongly with hemorrhagic fraction 8; CA-5+ had a broader reactivity and reacted with several HPLC hemorrhagic and non-hemorrhagic fractions. Crude venoms of C. adamanteus, C. scutulatus scutulatus and C. viridis lutosus reacted with CA-P-8, while C. viridis lutosus, C. viridis oreganus, C. scutulatus scutulatus and C. horridus horridus reacted with CA-5+. C. molossus molossus and C. lepidus lepidus did not react with CA-P-8 and CA-5+. Hemorrhagic HPLC fractions 6, 7, 8, were completely neutralized by monoclonal antibody CA-P-8; fraction 9 was partially neutralized. The present study indicated that some C. atrox venom HPLC hemorrhagic fractions have both common and unique epitopes. Antigenic determinants were also found to be shared among different Crotalus species.(ABSTRACT TRUNCATED AT 250 WORDS)


MRI abnormalities in Creutzfeldt-Jakob disease.
Two patients with biopsy-proven Creutzfeldt-Jakob disease had MRI studies that revealed increased signal in the basal ganglia on T-2 weighted images, suggesting that MRI can be a useful diagnostic instrument in Creutzfeldt-Jakob disease.


Minimizing Low Back Cumulative Loading during Design of Manual Material Handling Tasks: An Optimization Approach.
OCCUPATIONAL APPLICATIONSWe present a practical method for minimizing low-back cumulative loading that leverages digital human modeling capabilities and optimization using an evolutionary algorithm. We demonstrate use of the method in a simulated lifting task. Our results show that this method is robust to different routines for calculating cumulative loading. The proposed method can aid ergonomics engineers in addressing a potential risk factor early in the design stage, even in the absence of an established threshold limit value, and it provides a time saving by eliminating the need to adjust workplace parameters across many design possibilities.


Lymphoma presenting as parotid xanthogranulomatous sialadenitis.
We present a case of disseminated high-grade, T-cell-rich B-cell lymphoma in a 72-year-old white woman that presented as unilateral parotid swelling, and was initially diagnosed as xanthogranulomatous sialadenitis.


Topotecan (Hycamtin) responsiveness in human renal carcinoma cell lines of the clear cell and papillary types.
Since no effective therapeutic approach is yet known for metastatic renal cell carcinoma (RCC), we analyzed the effects of topotecan (Hycamtin), a novel topoisomerase I-inhibitor, in RCC cell lines of the clear cell and papillary/chromophilic types. The anti-proliferative and apoptosis-inducing effects of topotecan were analyzed in 20 RCC cell lines by MTT-assay and light microscopic apoptosis counting. Moreover, Bcl-2 and Bax expression was investigated by Northern blot and immunocytochemistry while the p53 mutation status was analyzed by DNA sequencing. Exposure to clinically relevant concentrations of topotecan (i.e. < or = 1 microg/ml) resulted in a significant (p<0.05) dose-dependent reduction of cell number in 17 out of 20 RCC cell lines. The reduction of cell number was paralleled by an increase in apoptotic cell death. Papillary/chromophilic RCCs exhibited a significantly (p<0.05) more pronounced responsiveness to topotecan than clear cell RCCs. Moreover, the effects of topotecan proved to be superior to those of 5-fluorouracil (5-FU), an anticancer drug currently used in the therapy of RCCs. No correlation became evident between responsiveness to topotecan and the expression levels of Bcl-2 and Bax. Moreover, the response to topotecan could not be correlated with the p53 mutation status of our RCC cell lines. Clinically relevant concentrations of topotecan induced apoptosis in RCC cell lines more effectively than 5-FU. Further testing will show whether topotecan-induced apoptosis can be exploited for the treatment of RCCs in vivo as well.


Asymmetric ligand recognition by the activating natural killer cell receptor NKG2D, a symmetric homodimer.
Natural killer (NK) cells function through a diverse array of cell-surface natural killer receptors (NCRs). NCRs specific for classical and non-classical MHC class I proteins, expressed in complex patterns of inhibitory and activating isoforms on overlapping, but distinct, subsets of NK cells, play an important role in immunosurveillance against cells that have reduced MHC class I expression as a result of infection or transformation. Another NCR, NKG2D, is an activating NCR first identified on NK cells, but subsequently found on macrophages and a variety of T cell types. NKG2D ligands in rodents include the MHC class I-like proteins RAE-1 and H60 and, in humans, ULBPs and the cell stress-inducible proteins MICA and MICB. NKG2D-MIC and -RAE-1 recognition events have been implicated in anti-viral and -tumor immune responses. Crystallographic analyses of NKG2D-MICA and -RAE-1 complexes reveal an unusual mode of recognition that apparently tolerates a surprising degree of ligand plasticity while generating affinities that are among the strongest TCR- or NCR-ligand affinities, thus, far described.


Construction of a family of biphenyl combinatorial libraries: structure-activity studies utilizing libraries of mixtures.
A set of biphenyl aminoacid building blocks has been synthesized. These were used to construct partially-peptidic combinatorial libraries as equimolar multi-component samples. Activity of members of this library as vitronectin receptor antagonists is described, together with SAR studies of the most active members. These studies illustrate several important features of combinatorial libraries.


Mechanical Adaptability of Tumor Cells in Metastasis.
The most dangerous aspect of cancer lies in metastatic progression. Tumor cells will successfully form life-threatening metastases when they undergo sequential steps along a journey from the primary tumor to distant organs. From a biomechanics standpoint, growth, invasion, intravasation, circulation, arrest/adhesion, and extravasation of tumor cells demand particular cell-mechanical properties in order to survive and complete the metastatic cascade. With metastatic cells usually being softer than their non-malignant counterparts, high deformability for both the cell and its nucleus is thought to offer a significant advantage for metastatic potential. However, it is still unclear whether there is a finely tuned but fixed mechanical state that accommodates all mechanical features required for survival throughout the cascade or whether tumor cells need to dynamically refine their properties and intracellular components at each new step encountered. Here, we review the various mechanical requirements successful cancer cells might need to fulfill along their journey and speculate on the possibility that they dynamically adapt their properties accordingly. The mechanical signature of a successful cancer cell might actually be its ability to adapt to the successive microenvironmental constraints along the different steps of the journey.


Tumor immune microenvironment and immune checkpoint inhibitors in esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC) is the main prevalent histological type of esophageal cancer, predominantly constituting 90% of cases worldwide. Despite the development of multidisciplinary therapeutic approaches, its prognosis remains unfavorable. Recently, the development of monoclonal antibodies inhibiting programmed death 1 (PD-1) or programmed death-ligand 1 (PD-L1) has led to marked therapeutic responses among multiple malignancies including ESCC. However, only a few patients achieved clinical benefits due to resistance. Therefore, precise and accurate predictive biomarkers should be identified for personalized immunotherapy in clinical settings. Because the tumor immune microenvironment can potentially influence the patient's response to immune checkpoint inhibitors, tumor immunity, such as PD-L1 expression on tumors, tumor-infiltrating lymphocytes, tumor-associated macrophages, and myeloid-derived suppressor cells, in ESCC should be further investigated. In this review, accumulated evidence regarding the tumor immune microenvironment and immune checkpoint inhibitors in ESCC are summarized.


Construct validity and psychometric properties of the Chinese version of the City Birth Trauma Scale.
Currently, most postpartum posttraumatic stress disorder (PTSD) screening scales used in China are general PTSD scales which are not compiled specifically for pregnant women and thus cannot reflect the unique needs of this population. This study aimed to translate the City Birth Trauma Scale (City BiTS) into Chinese and validate its psychometric characteristics in Chinese postpartum women. After translation, back-translation, and expert discussion, 596 mothers at 1 to 12 months postpartum filled out the questionnaires through the Internet. The reliability and validity of the translated questionnaire were tested. The Cronbach's α coefficient of the Chinese version of City BiTS (City BiTS-C) was 0.889, the test-retest reliability was 0.86, and the content validity was 0.93. Exploratory factor analysis extracted two factors accounted for 63.148% of the variance. The City BiTS-C had appropriate construct validity in the Chinese culture (root mean square error of approximation [RMSEA] = 0.048, <0.05; χ2 /df = 2.666, <3). The values of the incremental fit index (IFI) and the Tucker-Lewis coefficient (TLI) were 0.990 and 0.976, which identified that the model was a good fit for the data. The values of the comparative fit index (CFI) and the normed fit index (NFI) were 0.890 and 0.873, respectively. The City BiTS-C is a reliable and valid measure to screening and diagnosis the postpartum PTSD among new mothers who gave birth in the past year in mainland China. The City BiTS-C is a short, reliable, and valid instrument that measures the symptoms of postpartum PTSD, and it is recommend for clinical screening.


Two-dose strategies for human papillomavirus vaccination: how well do they need to protect?
Two-dose human papillomavirus (HPV) vaccine schedules may provide short-term protection but their long-term population impact is unknown. Two models of HPV transmission and associated cervical disease (squamous and glandular, neoplasia and cancer) were fitted to data from England and Canada on HPV epidemiology, sexual behaviour, cervical screening outcomes and cervical cancer incidence. Models suggest that at 40-80% coverage, if two-dose schedules protect vaccinees for 20 years, then the benefits of the third dose are small. If two doses protect for 10 years, then the third dose may prevent as many cancers as the first two. At 80% coverage, numbers needed to receive a third dose to prevent an additional cancer are 5900-110,000 (England), 3000-5100 (Canada) with 20 years two-dose protection, and 2000-5300 (England), 760-950 (Canada) with 10 years two-dose protection. Results enable decision makers to quantify risks associated with two-dose schedules despite remaining uncertainties in vaccine duration and cross-protection.


Effects of oral bisphosphonates on myopic choroidal neovascularisation over 2 years of follow-up: comparison with anti-VEGF therapy and photodynamic therapy. A pilot study.
Choroidal neovascularisation is often associated with pathological myopia. Bisphosphonates (BP), the preferred drug for treatment of osteoporosis, are known to have anti-angiogenic effects. To compare the therapeutic effects of oral BP with anti-vascular endothelial growth factor therapy (anti-VEGF) and photodynamic therapy (PDT) for myopic choroidal neovascularisation (mCNV) over 2 years of follow-up. One hundred eyes of 96 consecutive patients with mCNV who underwent oral BP treatment (alendronate 5 mg/day or 35 mg/week), anti-VEGF therapy, PDT or observation only were followed up for 2 years. The best-corrected visual acuity (BCVA) and the central retinal thickness (CRT) in optical coherence tomography were compared among the treatment groups. The mean BCVA of the patients was maintained for up to 2 years in the BP and PDT groups. In the anti-VEGF group, the mean BCVA was significantly improved but was significantly worse in the no-treatment group. The visual outcomes were significantly better in the BP, PDT and anti-VEGF groups than the no-treatment group over 2 years of follow-up (-0.28, -0.26 and -0.39 logMAR units, p=0.032, 0.021 and 0.0004, respectively). The mean CRT was significantly decreased in all treatment groups (-84, -121 and -122 mm, p=0.0025, 0.017 and 0.000025, respectively). Oral BP should be investigated further as possible therapeutic and preventive drugs for mCNV.


Developmental implications of early-life otitis media.
This symposium addressed possible relationships between early-life otitis media (OM) and various components of child development. The existence and nature of such relationships are of substantial theoretical and practical consequence. Particularly at issue are policies concerning optimal frequencies of examination and screening for middle-ear disease as well as the relative appropriateness of conservative vs. aggressive regimens of management.


Severe low-titer cold-hemagglutinin disease responsive to steroid pulse therapy.
We report the case of a 12-year-old boy suffering from severe low-titer cold-hemagglutinin disease with excruciating colicky abdominal pain, jaundice, and acute hemolytic anemia requiring transfusion. Cold hemagglutinins of the IgM type and a positive direct antiglobulin test, predominantly against C3d, were found. Steroid pulse therapy with 20 mg/kg body wt. methylprednisolone for 3 consecutive days was given. Abdominal pain disappeared within 12 h of the first steroid infusion and hemolysis was halted. We conclude that a therapeutic trial with steroid pulse therapy in severe low-titer cold-hemagglutinin disease is warranted.


The use of Tween 20 alone as a blocking agent for immunoblotting can cause artefactual results.
The use of Tween 20 as a suitable blocking agent in immunoblotting studies was evaluated by screening a panel of monoclonal antibodies (MoAbs) against a selection of blotted proteins which were unrelated to the antigens used to raise the MoAbs. Using Tween 20 alone to block the nitrocellulose membranes clear reactions were observed between the panel of MoAbs and several components of the blotted protein mixture. In contrast, when haemoglobin was used to block the membranes such reactions were not observed. In the absence of added protein the use of Tween 20 alone as a blocking agent for immunoblotting appears to lead to false positive reactions by non-specific antigen-antibody complexes.


Usefulness of new diastolic strain and strain rate indexes for the estimation of left ventricular filling pressure.
Tissue Doppler indexes of left ventricular (LV) filling pressure are prone to angulation errors and tethering and are less reliable in patients with preserved LV ejection fraction and indeterminate early peak transmitral diastolic flow (E)/mitral early diastolic velocity (Ea) (8 <E/Ea <15). Two-dimensional echocardiographic global longitudinal diastolic strain (Ds) and strain rate (DSr) were measured during peak mitral filling, and combined with E, derived new noninvasive indexes of LV filling pressure (E/Ds and E/10DSr). These indexes were compared with simultaneously invasively measured LV preatrial (pre-A) contraction pressure and E/Ea. Fifty patients were studied. Mean age was 55.9 +/- 9.9 years, 22 (43%) were women, and mean LV ejection fraction was 49.3 +/- 18.0%. Ds (R = 0.48, p <0.001) and DSr (R = 0.43, p = 0.002) correlated with invasively measured -dP/dt. Correlations between E/Ds and E/10DSr with LV pre-A pressure were R = 0.81 (p <0.001) and R = 0.80 (p <0.001) compared with R = 0.63 (p <0.001) between E/Ea and LV pre-A pressure, respectively. E/Ds > or =8 had higher sensitivity and specificity (95% and 94%, respectively; area under the curve = 0.96, p <0.0001) than E/Ea > or =15 (sensitivity 81%, specificity 75%; area under the curve = 0.85, p <0.0001) for the prediction of LV pre-A pressure > or =15 mm Hg (p = 0.01 for comparison). In patients with LV ejection fraction > or =50% and 8 <E/Ea <15, E/Ds and E/DSr were more accurate than E/Ea for determination of LV pre-A pressure. In conclusion, a novel ratio 2-dimensional echocardiographic diastolic strain ratio (E/Ds) was a better predictor of LV filling pressure than E/Ea. In patients with LV ejection fraction > or =50% or indeterminate E/Ea, both E/Ds and E/10DSr (a ratio based on global DSR) were better predictors of LV filling pressure than E/Ea.


Environmental impact of toxic elements in red mud studied by fractionation and speciation procedures.
Aluminum (Al) is mostly produced from bauxite ore, which contains up to 70% of Al(2)O(3) (alumina). Before alumina is refined to aluminum metal, it is purified by hot alkaline extraction. As a waste by-product red mud is formed. Due to its high alkalinity and large quantities, it represents a severe disposal problem. In Kidričevo (Slovenia), red mud was washed with water before disposal, and after drying, covered with soil. In Ajka (Hungary), the red mud slurry was collected directly in a containment structure, which burst caused a major accident in October 2010. In the present work the environmental impact of toxic elements in red mud from Kidričevo and Ajka were evaluated by applying a sequential extraction procedure and speciation analysis. The predominant red mud fraction was the insoluble residue; nevertheless, environmental concern was focused on the highly mobile water-soluble fraction of Al and Cr. Al in the water-soluble Ajka mud fraction was present exclusively in form of toxic [Al(OH)(4)](-), while Cr existed in its toxic hexavalent form. Comparative assessment to red mud from Kidričevo (Slovenia) with a lower alkalinity (pH 9) with that from Ajka demonstrated significantly lower Al solubility and the presence of only trace amounts of Cr(VI), confirming that disposal of neutralized mud is environmentally much more acceptable and carries a smaller risk of ecological accidents. Since during the Ajka accident huge amounts of biologically available Al and moderate Cr(VI) concentrations were released into the terrestrial and aquatic environments, monitoring of Al and Cr(VI) set free during remedial actions at the contaminated site is essential. Particular care should be taken to minimize the risk of release of soluble Al species and Cr(VI) into water supplies and surface waters.


Comparing the feasibility and accuracy of three-dimensional ultrasound to two-dimensional ultrasound and computed tomography angiography in the assessment of carotid atherosclerosis.
Two-dimensional ultrasound (2D-US) is the mainstay imaging technique used to evaluate carotid atherosclerosis. An automated single sweep three-dimensional ultrasound (3D-US) technique became available. We evaluated the feasibility and accuracy of 3D-US in the assessment of carotid plaques compared to those of 2D-US. Carotid computed tomography angiography (CTA) was used as a reference. Among 126 stroke patients who underwent carotid 2D-US, 73 underwent 3D-US and carotid CTA. 3D-US was pursued when there were carotid plaques or when area stenosis was ≥ 20% by 2D-US. Both 2D- and 3D-US images of the carotid arteries were acquired using a dedicated ultrasound system that was equipped with the single sweep volumetric transducer. In total, 266 arteries from 73 patients were selected for comparison of the detection rate of carotid plaques between 2D- and 3D-US. Among the 73 patients, carotid CTA detected 139 plaques. 3D-US demonstrated a higher detection rate of carotid plaques than did 2D-US (108 plaques (77.9%) vs. 70 plaques (50.4%)) when using carotid CTA as a reference standard. Carotid plaque volume (PV) of 133 vessels from 73 patients were quantitatively evaluated using both 3D-US and carotid CTA. Plaque volume of carotid artery was comparable between 3D-US and CTA (148.5 ± 133.0 mm3 vs. 154.1 ± 134.6 mm3 , P = .998, R: 0.9825, P-value for r < .001). 3D-US using a single sweep technique was a feasible and accurate method of detecting arterial plaques and assessing plaque volume.


Prognostic significance of the infiltrative pattern invasion in endometrioid adenocarcinoma of the endometrium.
The prognostic significance of the invasive type of carcinoma cells in endometrial carcinoma is not defined. We evaluated the prognostic significance of the invasive type, as well as the immunostains of p53, c-erbB-2, Ki-67 antigen and MDM2 in endometrial endometrioid adenocarcinoma. This prospective analysis comprised 112 patients with endometrioid adenocarcinoma of the uterine corpus who had undergone surgery and were traced for more than 5 years after the operation. They were divided into recurrence (16 patients) and non-recurrence (96 patients) groups. The invasive type of carcinoma cells was divided into expansile, mixed (expansile and infiltrative) and infiltrative pattern. The difference in the invasive type (P < 0.001) and p53 expression (P = 0.004) between the recurrence and non-recurrence groups was significant in the univariate analysis. Moreover, the invasive type was significant in the multivariate analysis (P = 0.004). In contrast, the difference in MDM2 expression, c-erbB-2 expression and the Ki-67 labeling index in both groups was not significant in the univariate analysis. The infiltrative pattern of the invasive type (P < 0.001) and p53 expression (P = 0.043) were significantly related to a poor prognosis in the Kaplan-Meier method using the log-rank test. In conclusion, the current study indicated that the infiltrative pattern of the carcinoma cells is a predictor for poor prognosis in endometrioid adenocarcinoma in the uterine corpus. It was also indicated that p53 immunostains are useful as a predictor, but Ki-67 antigen, c-erbB-2 and MDM2 stains are not.


Tooth agenesis patterns and variants in PAX9: A systematic review.
Mutations in PAX9 are the most common genetic cause of tooth agenesis (TA). The aim of this study was to systematically review the profiles of the TA and PAX9 variants and establish their genotype-phenotype correlation. Forty articles were eligible for 178 patients and 61 mutations (26 in frame and 32 null mutations). PAX9 mutations predominantly affected molars, mostly the second molar, and the mandibular first premolar was the least affected. More missing teeth were found in the maxilla than the mandible, and with null mutations than in-frame mutations. The number of missing teeth was correlated with the locations of the in-frame mutations with the C-terminus mutations demonstrating the fewest missing teeth. The null mutation location did not influence the number of missing teeth. Null mutations in all locations predominantly affected molars. For the in-frame mutations, a missing second molar was commonly associated with mutations in the highly conserved paired DNA-binding domain, particularly the linking peptide (100% prevalence). In contrast, C-terminus mutations were rarely associated with missing second molars and anterior teeth, but were commonly related to an absent second premolar. These finding indicate that the mutation type and position contribute to different degrees of loss of PAX9 function that further differentially influences the manifestations of TA. This study provides novel information on the correlation of the PAX9 genotype-phenotype, aiding in the genetic counseling for TA.


Safety and immunogenicity of recombinant poxvirus HIV-1 vaccines in young adults on highly active antiretroviral therapy.
A trial to evaluate the safety and immunogenicity of recombinant modified vaccinia Ankara (MVA) and fowlpox (FP) vectors expressing multiple HIV-1 proteins was conducted in twenty HIV-1 infected youth with suppressed viral replication on HAART. The MVA and FP-based multigene HIV-1 vaccines were safe and well tolerated. Increased frequencies of HIV-1 specific CD4+ proliferative responses and cytokine secreting cells were detected following immunization. Increased frequencies and breadth of HIV-1 specific CD8 T-cell responses were also detected. Plasma HIV-1-specific antibody levels and neutralizing activity were unchanged following vaccination. Poxvirus-based vaccines may merit further study in therapeutic vaccine protocols.


Economic evaluations of health care interventions in oropharyngeal dysphagia after stroke: protocol for a systematic review.
Oropharyngeal dysphagia (OD) affects 40-81% of patients after stroke. A recent systematic review on the costs of OD and its main complications showed higher acute and long-term costs for those patients who developed OD, malnutrition and pneumonia after stroke. These results suggest that appropriate management of post-stroke OD could reduce clinical complications and costs. The purpose of this systematic review is to assess the available literature for healthcare interventions that are efficient or cost-effective in the management of OD. A systematic review on economic evaluations of health care interventions will be performed on post-stroke patients with OD following PRISMA recommendations. Four bibliographic databases will be searched and a subsequent reference check will be done. English and Spanish literature will be included without date restrictions. Studies will be included if they refer to economic evaluations or in which cost savings were reported in post-stroke patients suffering OD. Studies will be excluded if they are partial economic evaluation studies, if they refer to esophageal dysphagia, or if OD is caused by causes different from stroke. Evidence will be presented and synthetised with a narrative method and using tables. Quality evaluation will be done using the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) statement. The protocol for this systematic review is the first step to assess the cost-effectiveness of the healthcare interventions that have been described as potential treatments for post-stroke OD. This systematic review will summarise the current evidence on the relation between cost and benefits associated with the appropriate management of OD in post-stroke patients. PROSPERO CRD42020136245.


Reaction of substrates with 35S-thiophosphorylated succinyl-CoA synthetase of pig heart. Similarities to the case of the Escherichia coli enzyme.
Guanosine 5'-O-(3-thio)triphosphate (GTP gamma S) was found to be a substrate of pig heart succinyl-CoA synthetase with Km and kcat values of 3 microM and 0.23 s-1, respectively. The corresponding values with GTP as substrate were 48 microM and 65 s-1. 35S-thiophosphorylated enzyme was prepared by incubation of pig heart succinyl-CoA synthetase with [35S]GTP gamma S. A comparison was made of thiophosphoryl group release by substrates from this alpha beta (one active site) enzyme with that of the alpha 2 beta 2 (two active sites) Escherichia coli enzyme (Wolodko, W. T., Brownie, E. R., O'Connor, M. D., and Bridger, W. A. (1983) J. Biol. Chem. 258, 14116-14119; Nishimura, J. S., and Mitchell, T. (1984) J. Biol. Chem. 259, 9642-9645). It was found, as in the case of the E. coli enzyme, that thiophosphoryl group release by GDP and by succinate plus CoA was stimulated by succinyl-CoA and GTP, respectively. The same result was observed at 1, 0.1, and 0.01 mg/ml, lending assurance that these phenomena were not exhibited by an aggregated form of the pig heart enzyme. While an alternating-sites catalytic cooperativity model is not ruled out for the E. coli enzyme, it is proposed that the NTP- and succinyl-CoA-stimulated release of thiophosphoryl groups from either enzyme involves a "same-site" mechanism, to be distinguished from an "other-site" mechanism.


Initial Experience with Surpass Evolve Flow Diverter in the Treatment of Intracranial Aneurysms.
The principle of flow diversion has revolutionized the treatment of brain aneurysms. In this study, we report our experience of the new Surpass Evolve (SE) flow diverter in the treatment of intracranial aneurysms. Patients were treated with the SE as first-line therapy between May 2019 and June 2020 at 2 experienced institutions. Inclusion criteria were wide-necked, blister-like, or fusiform/dissecting aneurysms in the anterior and posterior circulation. Primary endpoint was technical success defined as favorable navigation to the target vessel and successful deployment of the SE. Secondary endpoints were favorable aneurysm occlusion defined as O'Kelly Marotta (OKM) scale C1-3 + D on follow-up, procedure-related complications and retreatment. A total of 46 aneurysms in 42 patients were treated with 57 SE flow diverters. Median aneurysm size was 6.6 mm (IQR 4.0-12.2 mm) with a median neck width of 4 mm (IQR 2.2-5.4 mm). On admission, 6 (13%) aneurysms were ruptured and 41 (89%) were located in the anterior circulation. The primary endpoint was reached in 96%. Median follow-up was 116 days (IQR 92-134 days) and available for 36/46 (78%) aneurysms. Favorable aneurysm occlusion was seen in 31/36 (86%) aneurysms and 27/36 (75%) were occluded completely. Parent artery occlusion appeared in 3 (3%) patients on follow-up and 2 aneurysms (6%) required additional treatment due to insufficient closure. The new SE flow diverter is safe and seems to be effective with promising occlusion rates at short-term follow-up.


[Child health in the COVID-19 pandemic: results from school entry data and a parent survey in the Hanover region].
With its social contact restrictions, the COVID-19 pandemic can be very burdensome for children and adolescents. The regularly conducted school entry examination in Germany delivers data on the development and health of pre-school children and can deliver indications on the impacts of the pandemic. Based on school entry data for the Hannover region, it will be examined if there are more children with noticeable development and health impairment after the first lockdown in 2020 compared to before the COVID-19 pandemic. The prevalence of the results of the 2017/18-2020/21 school entry cohorts are compared. A survey of parents presents information of the everyday life and wellbeing of children and their families during the pandemic. After the first lockdown phase, the data shows a significant increase of noticeable language problems and an increase of children with a need for language support and with inadequate or no German language knowledge in comparison to the years before. Further, an increase of overweight or obesity as well as an increase in noticeable fine motor skill abnormalities and media consumption problems could be observed. The survey shows that the share of children who are more often sad increased to 32.1%, and the share of children who have more frequent rage attacks increased to 24.9%. Children more frequently have problems falling asleep and staying asleep or have stomachaches, headaches, sickness, or loss of appetite. These results are in line with other current studies and provide evidence of the possible effects of the COVID-19 pandemic on children's health. However further studies are necessary in order to deliver statistically firm results.


Brief evaluation of pleasantness of olfactory and trigeminal stimulants.
To evaluate a brief olfactory test based on pleasantness rating of olfactory and trigeminal stimulants. Criterion standard. University hospital. A total of 60 participants (48 healthy and 12 with olfactory loss) aged 16 to 81 years. The new test of odor pleasantness (TOP) based on classification of odorants and trigeminal stimulants into 4 categories was compared with 2 standard tests of subjective olfactometry. We evaluated the possible use of the TOP in assessment of olfactory and the trigeminal system in healthy individuals and those with olfactory loss. All 3 tests demonstrated significant mutual correlation (P < .01), and persons with olfactory loss scored lower than healthy participants (P < .01). Using exploratory factor analysis and cluster analysis, we found that healthy individuals classified odorants accurately according to the degree of their hedonic character. The TOP offers outcomes similar to those of standard tests of olfaction that are based on psychophysical testing.


Vascular endothelial growth factor in colorectal cancer.
Angiogenesis plays an important role in colorectal cancer progression. Evidence from preclinical and clinical studies indicates that vascular endothelial growth factor (VEGF) is the predominant angiogenic factor in human colorectal cancer and is associated with formation of metastases and poor prognosis. Based on these results it was hypothesized that attacking one or more of the VEGF-mediated mechanisms may be promising in the treatment of colorectal cancer. AIMS. This article reviews the role of VEGF in colon cancer and summarizes recent advances in the treatment of colorectal cancer by anti-VEGF strategies.


miR-1306-3p targets FBXL5 to promote metastasis of hepatocellular carcinoma through suppressing snail degradation.
This study aimed to elucidate the effect of miR-1306-3p on metastasis of hepatocellular carcinoma (HCC) and potential mechanism involved. miR-1306-3p promoted migration and invasion of HCC in vivo and in vitro. Moreover, miR-1306-3p inhibited snail to enhance its expression via directly targeting FBXL5, thus inducing the epithelial-mesenchymal transition (EMT) in HCC. Intriguingly, miR-1306-3p expression was transcriptionally enhanced by FoxM1. Consistently, miR-1306-3p was upregulated in HCC compared with paracarcinoma and correlated with poor prognosis of HCC patients. Our researches suggest that miR-1306-3p is a tumor enhancer in regulating of HCC metastasis, and miR-1306-3p may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.


3-trifluoromethylcarbacephems: synthesis of broad spectrum antibacterial compounds.
The enhanced stability of the carbacephem nucleus over the corresponding cephalosporin nucleus has allowed the synthesis of 7-arylglycyl-3-trifluoromethyl-carbacephems. These unique carbacephems possess broad spectrum activity and high stability to both plasmid and chromosomally mediated beta-lactamases.


Precise CRISPR-Cas-mediated gene repair with minimal off-target and unintended on-target mutations in human hematopoietic stem cells.
While CRISPR-Cas9 is key for the development of gene therapy, its potential off-target mutations are still a major concern. Here, we establish a "spacer-nick" gene correction approach that combines the Cas9D10A nickase with a pair of PAM-out sgRNAs at a distance of 200 to 350 bp. In combination with adeno-associated virus (AAV) serotype 6 template delivery, our approach led to efficient HDR in human hematopoietic stem and progenitor cells (HSPCs including long-term HSCs) and T cells, with minimal NHEJ-mediated on-target mutations. Using spacer-nick, we developed an approach to repair disease-causing mutations occurring in the HBB, ELANE, IL7R, and PRF1 genes. We achieved gene correction efficiencies of 20 to 50% with minimal NHEJ-mediated on-target mutations. On the basis of in-depth off-target assessment, frequent unintended genetic alterations induced by classical CRISPR-Cas9 were significantly reduced or absent in the HSPCs treated with spacer-nick. Thus, the spacer-nick gene correction approach provides improved safety and suitability for gene therapy.


The regulation mechanism of HLA class II gene expression at the level of mRNA stability.
The number of major histocompatibility complex (MHC) class II antigens may be regulated at different levels. Although transcriptional regulation has been studied most intensely, evidence for control mechanisms acting on the stability of MHC class II mRNAs has been reported. We have previously shown, in fact, that the half-life of MHC class II mRNA rapidly decreases in Raji cells upon inhibition of translation by cycloheximide; further data indicated that this effect was not correlated with the inhibition of the synthesis of trans-acting protein(s) required for mRNA stability. In the present work, we developed an in vitro mRNA decay assay system to measure HLA-DRA mRNA stability and used inhibitors of protein synthesis affecting different steps of the process of translation in order to discriminate among possible mechanisms determining controlled MHC class II mRNA hydrolysis. We found that HLA-DRA mRNA associated with polysomes derived from cells treated with either puromycin (which causes dispersion of polysomes and accumulation of monosomes) or cycloheximide (which slows down translation causing ribosome stalling) is more rapidly degraded than in the absence of protein synthesis inhibitors. On the basis of our findings, we suggest that arrest of protein synthesis per se exposes the HLA-DRA mRNA molecules to degradative activities co-sedimenting with the polysomal fraction.


Task variables determine which biological clock controls circadian rhythms in human performance.
There are circadian (approximately 24 h) rhythms for a wide range of human physiological and psychological functions including mood and performance efficiency. These rhythms are self-sustaining in conditions of temporal isolation, indicating that internal oscillators (or biological clocks) control them. Recent research has proposed an endogenous two-oscillator model of the human circadian system, with one oscillator indicated by the core body temperature rhythm and a second oscillator responsible for the daily cycle of sleep and wakefulness. The present study was designed to produce a desynchronization in period between the two oscillators, to assess directly the behaviour of the rhythms of different performance tasks. The results, reported here, indicate that a simple manual dexterity task is almost entirely under the control of the temperature rhythm oscillator, whereas a more complex cognitive task demonstrates a periodicity which appears to be influenced by those oscillators controlling temperature and the sleep/wake cycle.


Distinct regulation of vasoactive intestinal peptide (VIP) expression at mRNA and peptide levels in human neuroblastoma cells.
Neuronal differentiation was induced in cultures of the human neuroblastoma cell line subclone SH-SY5Y by 14-day treatment with dibutyryl cAMP (dBcAMP), retinoic acid, and phorbol 12-myristate 13-acetate (PMA). An approximate 4-fold increase in vasoactive intestinal peptide (VIP) mRNA concentration was observed after differentiation with retinoic acid, whereas no change in VIP mRNA concentration was observed after differentiation with dBcAMP or PMA. A short-term treatment of cells with PMA did however result in a 5-fold transient increase in VIP mRNA; prior differentiation with retinoic acid or dBcAMP diminished this effect. Observed increases in VIP mRNA were in all cases accompanied by increases in VIP immunoreactivity. Remarkably, however, long-term treatment of cells with dBcAMP, which caused no change in mRNA levels, resulted in a six-fold increase in VIP immunoreactivity. Acute (36-h) treatment with carbachol also caused an increase in VIP immunoreactivity (about 2-fold, and blocked by atropine) without an increase in VIP mRNA level. Thus, a quantitative change in gene transcription or mRNA stability appears not to be a prerequisite for increased VIP expression, indicating that regulation can occur at translational or post-translational steps.


Changes in the adolescent brain and the pathophysiology of psychotic disorders.
Adolescence is a time of extensive neuroanatomical, functional, and chemical reorganisation of the brain which parallels substantial maturational changes in cognition and affect regulation. This period is characterised by stabilisation of synapses to diminish redundancy and increase efficiency of neural function, fine-tuning of excitatory and inhibitory neurotransmitter systems, beginning of integration between late maturing and early maturing brain structures, and development of effective connections. In effect, these so-called moving parts create a state of dynamic change that might underlie adolescent behaviours. Imbalances or changes in timing of these developmental processes clearly increase the risk for psychiatric disorders. Genetic, environmental, and epigenetic factors that shape brain development and hormonal changes that affect stress reactivity could be reasons why some, but not all, adolescents are at a heightened risk of developing a psychopathological disorder. In this Series paper, we assess the neurobiology of the changing adolescent brain, implications of this knowledge, and future research in major psychiatric disorders, particularly for psychotic disorders.


Distribution of transglutaminase family members in mouse whole body sections.
Transglutaminases (TGs) comprise a protein family in which the members catalyze the formation of isopeptide bonds between glutamine and lysine residues in various proteins. Eight enzymes have been identified and designated as factor XIII (FXIII) and TG1-7. Expression studies of four major members, i.e., FXIII, TG1, TG2, and TG3, have been performed in a relatively large number of mammalian tissues in comparison with those on the other isozymes. The structural and biochemical characteristics of these individual isozymes and expression analyses of TG family in some tissue extracts have been reported, but there have been no simultaneous comparative analyses of both their mRNA and protein expression patterns in tissues distributions. Thus, we developed novel experimental systems for in situ hybridization using cryofilm attached to whole body sections of neonatal mice, thereby obtaining data regarding the tissue distributions of the major TG isozymes. In this study, we performed the first detailed comparative analysis of the mRNA and protein distribution studies of TG family members in a wide range of mouse tissues. These data will be helpful for elucidating the unknown physiological and pathological functions of TGs.


Cyclothiazide reverses AMPA receptor antagonism of the 2,3-benzodiazepine, GYKI 53655.
On rat cortical slices, cyclothiazide, 1-100 microM, (ED50 = 7.1 +/- 1.1 microM) enhanced the depolarizing action of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) but not that of N-methyl-D-aspartate (NMDA). Cyclothiazide 10 microM also reversed the action of a 2,3-benzodiazepine, GYKI 53655, which is a non-competitive AMPA receptor antagonist, but not that of the quinoxalinedione, NBQX, which is a competitive AMPA receptor antagonist.


Assessment of virus infection in cultured cells using metabolic monitoring.
A rapid, in-process assessment of virus replication is disired to quickly investigate the effects of process parameters on virus infection, and to monitor consistency of process in routine manufacturing of viral vaccines. Live virus potency assays are generally based on plaque formation, cytopathic effect, or antigen production (TCID(50)) and can take days to weeks to complete. Interestingly, when infected with viruses, cultured cells undergo changes in cellular metabolism that can be easily measured. These phenomena appear to be common as they has been observed in a variety of virus-host systems, e.g., in insect cells infected with baculovirus, Vero cells infected with Rotavirus, MRC-5 cells infected with Hepatitis A virus, and MRC-5 cells infected with the Varicella Zoster Virus (VZV). In this article, changes in glycolytic metabolism of MRC-5 cells as a result of CVZ infection are described. Both glucose consumption and lactate production in VZV infected MRC-5 cells are significantly elevated in comparison to uninfected cells. Based on this result, a rapid, in-process assay to follow VZV infection has been developed. The relative increase in lactate production in infected cells (α) increases as the infection progresses and then plateaus as the infection peaks. This plateau correlates with time of peak virus titer and could be used as a harvest triggering parameter in a virus production process.X(u) = cell density of uninfected cellsX(i) = cell density of infected cellsX(T) = total cell densityL(i) = cumulative lactate production in infected culturesL(u) = cumulative lactate production in uninfected culturesq(Li) = specific lactate production of infected cellsq(Lu) = specific lactate production of uninfected cellsk(1), K(2) = constants.


What contributes to patient and parent satisfaction with medication in the treatment of children with ADHD? A report on the development of a new rating scale.
Satisfaction with medication is important in the evaluation of overall treatment outcome. There is a lack of consistent and validated rating scales for satisfaction with medication in ADHD, therefore comparison across studies is difficult. Here, we analyse the psychometric properties of the satisfaction with medication scale (SAMS), a new item-based questionnaire that assesses satisfaction with ADHD medication. Furthermore, we evaluate the predictive effect of ADHD symptoms and quality of life (QoL) on satisfaction. Data on satisfaction with Equasym XL(®) (methylphenidate) were collected in the OBSEER study using the parent (SAMS-P, n = 589) and patient (SAMS-S, n = 552) versions of the SAMS questionnaire. Internal consistency, item-total and cross-informant correlations, and the stability of satisfaction ratings over time were assessed. Satisfaction with medication scores were then correlated with ratings of ADHD symptoms and QoL. Rates of overall satisfaction with Equasym XL(®) among parents and children were high (>70%), as was internal consistency for both SAMS-P and SAMS-S (Cronbach's alpha > 0.9). Similarly, item-total correlations were high (r = 0.71-0.90) for SAMS-P and medium-high (r = 0.57-0.77) for SAMS-S. Cross-informant correlations and the stability of satisfaction ratings were moderate (r = 0.54-0.59 and 0.48-0.60, respectively). ADHD symptom and QoL ratings were significantly negative and positive predictors of satisfaction, explaining 36-52% of satisfaction variance at the final visit. The results show that parent and patient satisfaction was high and could be assessed reliably with the new SAMS questionnaire. Parent and patient ratings were moderately correlated, and symptom severity, functional impairment and QoL were the most significant predictors of satisfaction.


How does population agglomeration influence China's energy eco-efficiency? Evidence from spatial econometric analysis.
The contradiction between rapid urbanization and energy supply and demand is constraining the green development of Chinese cities. The impact of population agglomeration on urban energy eco-efficiency is still unclear. We used DMSP/OLS and NPP/VIIRS night lighting data, LandScan population dynamics statistics to extract "real urbanized areas" and "real urbanized populations" to measure the population agglomeration index. Considering the spatial interaction effects of economic behavior, we used panel data of 260 Chinese cities from 2006 to 2020 to construct a dynamic spatial Durbin model to explore the impact of population agglomeration on energy eco-efficiency. The study finds that population agglomeration contributes to regional energy eco-efficiency, with a spillover effect to surrounding areas. Endogeneity and robustness tests support the credibility of the results. Technological innovation plays a positive mediating role in the relationship between agglomeration and energy eco-efficiency. Heterogeneity analysis shows that the effect of population agglomeration on energy eco-efficiency varies across temporal and spatial dimensions. Our study provides valuable information for promoting urban energy eco-efficiency from the perspective of population agglomeration.


Low-loss hollow waveguide fibers for mid-infrared quantum cascade laser sensing applications.
We report on single mode optical transmission of hollow core glass waveguides (HWG) coupled with an external cavity mid-IR quantum cascade lasers (QCLs). The QCL mode results perfectly matched to the hybrid HE(11) waveguide mode and the higher losses TE-like modes have efficiently suppressed by the deposited inner dielectric coating. Optical losses down to 0.44 dB/m and output beam divergence of ~5 mrad were measured. Using a HGW fiber with internal core size of 300 µm we obtained single mode laser transmission at 10.54 µm and successful employed it in a quartz enhanced photoacoustic gas sensor setup.


The 5 A's of firearm safety counseling: Validating a clinical counseling methodology for firearms in a simulation-based randomized controlled trial.
The 5 A's of Firearm Safety Counseling is a novel framework by which clinicians can approach firearm injury prevention counseling. To evaluate this methodology as a tool for clinicians, a single-center, simulation-based randomized controlled trial was performed with clinical trainees in psychiatry, medicine, and pediatrics in an urban quaternary care center. Participants received didactic education on firearm injury epidemiology and evidence-based policies and training on a specific counseling framework, the 5 A's of Firearm Safety Counseling which they then implemented in a simulation setting with standardized patients. Of the 29 participants who were randomized, 28 completed the trial. Most participants were psychiatry trainees (residents or subspecialty fellows). While over 60% of participants were uncomfortable or extremely uncomfortable counseling on firearm injury prior to the interventions, only 4% reported being uncomfortable after receiving education and participating in simulated encounters. There was no significant difference between the quality and content of the counseling provided before and after the didactic-only session. There was a significant difference between the quality and content of the counseling provided before and after the specific training on the 5 A's for Firearm Safety Counseling strategy. The 5 A's for Firearm Safety Counseling is a promising educational tool to improve quality, content, and comfort delivering patient-centered counseling on firearm injury prevention in a simulation-based setting. These findings suggest that further validation in a clinical setting is warranted given there is an urgent need for feasible and effective firearm injury prevention strategies among clinicians.


Study of water mixing in the coastal waters of the western Taiwan Strait based on radium isotopes.
Radium is considered to be a useful tracer for studying the physical processes of seawater. In this work, three naturally occurring radium isotopes, (224)Raex, (226)Ra and (228)Ra, were measured in the coastal zone of the western Taiwan Strait during the summer seasons. Based on the distributions of the three radium isotopes and the salinity, we conclude that the water mixing pattern in the study area in summer consists of diluted water flowing from the Jiulong River to the open sea towards the east and southeast, and open sea seawater flowing inward from south to north. The submarine ground water discharges in the estuarine region, as suggested by the radium and salinity data. The residence times of the Jiulong River estuary, ranging from 7 to 49 d, were estimated using the radium isotope pairs (224)Raex and (226)Ra.


Automated MRI perfusion-diffusion mismatch estimation may be significantly different in individual patients when using different software packages.
To compare two established software applications in terms of apparent diffusion coefficient (ADC) lesion volumes, volume of critically hypoperfused brain tissue, and calculated volumes of perfusion-diffusion mismatch in brain MRI of patients with acute ischemic stroke. Brain MRI examinations of 81 patients with acute stroke due to large vessel occlusion of the anterior circulation were analyzed. The volume of hypoperfused brain tissue, ADC volume, and the volume of perfusion-diffusion mismatch were calculated automatically with two different software packages. The calculated parameters were compared quantitatively using formal statistics. Significant difference was found for the volume of hypoperfused tissue (median 91.0 ml vs. 102.2 ml; p < 0.05) and the ADC volume (median 30.0 ml vs. 23.9 ml; p < 0.05) between different software packages. The volume of the perfusion-diffusion mismatch differed significantly (median 47.0 ml vs. 67.2 ml; p < 0.05). Evaluation of the results on a single-subject basis revealed a mean absolute difference of 20.5 ml for hypoperfused tissue, 10.8 ml for ADC volumes, and 27.6 ml for mismatch volumes, respectively. Application of the DEFUSE 3 threshold of 70 ml infarction core would have resulted in dissenting treatment decisions in 6/81 (7.4%) patients. Volume segmentation in different software products may lead to significantly different results in the individual patient and may thus seriously influence the decision for or against mechanical thrombectomy. • Automated calculation of MRI perfusion-diffusion mismatch helps clinicians to apply inclusion and exclusion criteria derived from randomized trials. • Infarct volume segmentation plays a crucial role and lead to significantly different result for different computer programs. • Perfusion-diffusion mismatch estimation from different computer programs may influence the decision for or against mechanical thrombectomy.


Inflammation, sex, blood pressure changes and hypertension in midlife: the Hordaland Health Study.
Our aim was to test sex-specific associations of circulating markers of inflammation with blood pressure (BP) and incident hypertension in midlife. Participants in the Hordaland Health study (n = 3280, 56% women, mean age 48 years) were examined at baseline and followed for 6 years. Circulating levels of inflammatory markers including high-sensitive C-reactive protein (hs-CRP), neopterin, and pyridoxic acid ratio (PAr) index were measured at follow-up. The associations with systolic/diastolic BP and incident hypertension were tested in sex-specific linear- or logistic-regression analyses adjusted for body mass index, serum triglycerides, creatinine, physical activity, smoking and diabetes. At follow-up, women had lower mean BP than men (124/72 vs. 130/78 mmHg, p < 0.001). Higher hs-CRP was significantly associated with greater systolic and diastolic BP (standardized β = 0.07 and β = 0.09, both p < 0.01) in women, but not in men. Higher neopterin was associated with higher diastolic BP in women and higher PAr index was associated with higher diastolic BP in women and higher systolic and diastolic BP in men (all p < 0.01). Compared to hs-CRP < 1 mg/l, higher levels of hs-CRP 1-<3 mg/l and hs-CRP ≥ 3 mg/l were associated with new-onset hypertension only in women (odds ratio (OR) 1.74, 95% CI 1.20-2.53 and OR 1.87, 95% CI 1.20-2.90). Sex-interactions were found for hs-CRP and neopterin in models on incident hypertension and diastolic BP, respectively (both p < 0.05). Higher levels of circulating markers of inflammation were associated with higher BP and incident hypertension in a sex-specific manner. Our results suggest a sex-specific interaction between cardiovascular inflammation and BP in midlife.


Baseline anemia is not a predictor of all-cause mortality in outpatients with advanced heart failure or severe renal dysfunction. Results from the Norwegian Heart Failure Registry.
The aim of this study was to evaluate the prognostic impact of anemia in outpatients with chronic heart failure attending specialized heart failure clinics and specifically to investigate its prognostic utility in patients with severe renal dysfunction or advanced heart failure. Anemia is an independent prognostic marker in patients with heart failure. The effect of anemia on mortality decreases with increasing creatinine levels. Multivariate Cox regression analyses were used to investigate the prognostic effect of anemia in 4,144 patients with heart failure from 21 outpatient heart failure clinics in Norway. Severe renal failure was defined as estimated glomerular filtration rate ≤45 ml/min/1.73 m(2) and advanced heart failure as New York Heart Association functional classes IIIb and IV. Baseline anemia was present in 24% and was a strong predictor of all-cause mortality (adjusted hazard ratio [HR]: 1.30, 95% CI: 1.09 to 1.56, p = 0.004). Baseline anemia did not predict mortality in the 752 patients with severe renal dysfunction (adjusted HR: 1.08, 95 % CI: 0.77 to 1.51, p = 0.662) and the 528 patients with advanced heart failure (adjusted HR: 0.87, 95% CI: 0.56 to 1.34, p = 0.542). In the 1,743 patients who attended subsequent visits, sustained anemia independently predicted worse prognosis (adjusted HR: 1.47, 95% CI: 1.10 to 1.94, p = 0.008), whereas transient and new-onset anemia did not. According to our study, baseline anemia was not an independent predictor of all-cause mortality in outpatients with heart failure and accompanied severe renal dysfunction or advanced heart disease. Sustained anemia after optimizing heart failure treatment might imply worse prognosis independently of renal function and New York Heart Association functional class.


Optimal Reactive Power Dispatch in ADNs using DRL and the Impact of Its Various Settings and Environmental Changes.
Modern active distribution networks (ADNs) witness increasing complexities that require efforts in control practices, including optimal reactive power dispatch (ORPD). Deep reinforcement learning (DRL) is proposed to manage the network's reactive power by coordinating different resources, including distributed energy resources, to enhance performance. However, there is a lack of studies examining DRL elements' performance sensitivity. To this end, in this paper we examine the impact of various DRL reward representations and hyperparameters on the agent's learning performance when solving the ORPD problem for ADNs. We assess the agent's performance regarding accuracy and training time metrics, as well as critic estimate measures. Furthermore, different environmental changes are examined to study the DRL model's scalability by including other resources. Results show that compared to other representations, the complementary reward function exhibits improved performance in terms of power loss minimization and convergence time by 10-15% and 14-18%, respectively. Also, adequate agent performance is observed to be neighboring the best-suited value of each hyperparameter for the studied problem. In addition, scalability analysis depicts that increasing the number of possible action combinations in the action space by approximately nine times results in 1.7 times increase in the training time.


GABA-accumulating neurons in the nucleus raphe dorsalis and periaqueductal gray in the rat: a biochemical and radioautographic study.
The possibility of a GABAergic innervation of the nucleus raphe dorsalis (NRD) has been investigated by using the following approaches: (i) the identification of the principal neuronal groups afferent to the NDR by using horseradish peroxidase retrograde transport, (ii) the determination of glutamate decarboxylase activity (GAD) in the NRD after lesioning these groups or their putative pathways, and (iii) the radioautographic identification of terminals axons and nerve cells accumulating intraventricularly injected [3H]GABA. The hypothesis of a local GABAergic network is supported by the failure to obtain important changes in GAD after lesions of NRD afferents and the presence in this nucleus of terminals, fibers and nerve cell bodies accumulating [3H]GABA. It appears that these GABA-accumulating neurons could represent a portion of aperiventricular GABAergic system in the periaqueductal gray and the pontine ventricular gray.


The burden of heat-related stroke mortality under climate change scenarios in 22 East Asian cities.
Stroke is a leading cause of death and disability in East Asia. Owing to the aging population and high prevalence of stroke, East Asia might suffer a disproportionately heavy burden of stroke under the changing climate. However, the evidence relevant is still limited in this area. To evaluate the stroke mortality risk due to heat exposure in East Asia and predict its burden under various future climate change scenarios. We conducted a multi-center observational study and collected data from 22 representative cities in three main East Asian countries (i.e., China, Japan, and South Korea) from 1972 to 2015. The two-stage time-series analyses were applied to estimate the effects of heat on stroke mortality at the regional and country level. We further projected the burden of heat-related stroke mortality using 10 global climate models (GCMs) under four shared socioeconomic pathway and representative concentration pathway (SSP-RCP) scenarios, including SSP1-RCP1.9, SSP1-RCP2.6, SSP2-RCP4.5, and SSP5-RCP8.5 scenarios. In the present study, a total of 287,579 stroke deaths were collected during the warm season. Heat was significantly associated with an increased risk of stroke mortality. Overall, compared with the 2010 s, the heat-related attributable fraction (AF) was projected to increase in the 2090 s, with increments ranging from 0.8 % to 7.5 % across various climate change scenarios. The heat-related AF was projected to reach 11.9 % (95 % empirical confidence interval [eCI]: 6.1 %, 17.5 %) in the 2090 s under the SSP5-RCP8.5 scenario in China, while the corresponding estimates were 6.6 % (95 % eCI: 2.5 %, 11.0 %) and 5.1 % (95 % eCI: 1.2 %, 9.1 %) for Japan and South Korea, respectively. Climate change will exacerbate the burden of heat-related stroke mortality but with considerable geographical heterogeneity in East Asia.


Facile synthesis and application of poly(ionic liquid)-bonded silica hybrid materials.
Facile methods were developed to prepare hybrid poly(ionic liquid)-bonded silica for a wide range of applications, particularly in analytical chemistry. The hybrid material obtained was evaluated by comparing its adsorption capacity with other conventional separation materials. In addition, the hybrid material has the potential for industrial scale production.


BeEM: fast and faithful conversion of mmCIF format structure files to PDB format.
Although mmCIF is the current official format for deposition of protein and nucleic acid structures to the protein data bank (PDB) database, the legacy PDB format is still the primary supported format for many structural bioinformatics tools. Therefore, reliable software to convert mmCIF structure files to PDB files is needed. Unfortunately, existing conversion programs fail to correctly convert many mmCIF files, especially those with many atoms and/or long chain identifies. This study proposed BeEM, which converts any mmCIF format structure files to PDB format. BeEM conversion faithfully retains all atomic and chain information, including chain IDs with more than 2 characters, which are not supported by any existing mmCIF to PDB converters. The conversion speed of BeEM is at least ten times faster than existing converters such as MAXIT and Phenix. Part of the reason for the speed improvement is the avoidance of conversion between numerical values and text strings. BeEM is a fast and accurate tool for mmCIF-to-PDB format conversion, which is a common procedure in structural biology. The source code is available under the BSD licence at https://github.com/kad-ecoli/BeEM/ .


The yellow brick road to nuclear membrane mechanotransduction.
The nuclear membrane may function as a mechanosensory surface alongside the plasma membrane. In this Review, we discuss how this idea emerged, where it currently stands, and point out possible implications, without any claim of comprehensiveness.


Comparative proteomic analysis of mesenchymal stem cells derived from human bone marrow, umbilical cord and placenta: implication in the migration.
Umbilical cord (UC) and placenta (P) have been suggested as alternatives to bone marrow (BM) as sources of mesenchymal stem cells (MSC) for cell therapy, with both UC- and P-MSC possess immunophenotypic and functional characteristics similar to BM-MSC. However, under defined conditions, the migration capacity of BM- and P-MSC was found to be 5.9- and 3.2-folds higher than that of UC-MSC, respectively. By the use of 2-DE and combined MS and MS/MS analysis, six differentially expressed proteins were identified among these MSC samples, with five of them known to be involved in cell migration as migration enhancing or inhibiting proteins. Interestingly, the expression levels of those proteins reflect perfectly the migration capacity of corresponding MSC, which is also proved by in vitro overexpression and silencing techniques. Our study indicates that a bunch of migration-related proteins are pivotal in governing the migration capacity of MSC.


The tone atlas of perceptual discriminability and perceptual distance: Four tone languages and five language groups.
Some prior investigations suggest that tone perception is flexible, reasonably independent of native phonology, whereas others suggest it is constrained by native phonology. We address this issue in a systematic and comprehensive investigation of adult tone perception. Sampling from diverse tone and non-tone speaking communities, we tested discrimination of the three major tone systems (Cantonese, Thai, Mandarin) that dominate the tone perception literature, in relation to native language and language experience as well as stimulus variation (tone properties, presentation order, pitch cues) using linear mixed effect modelling and multidimensional scaling. There was an overall discrimination advantage for tone language speakers and for native tones. However, language- and tone-specific effects, and presentation order effects also emerged. Thus, over and above native phonology, stimulus variation exerts a powerful influence on tone discrimination. This study provides a tone atlas, a reference guide to inform empirical studies of tone sensitivity, both retrospectively and prospectively.


The oestrogen receptor regulates NFkappaB and AP-1 activity in a cell-specific manner.
Oestrogens regulate the expression of genes both positively and negatively in a range of cell types. These effects are mediated via the oestrogen receptor (ER) and involve direct interactions between the ER and DNA response elements, as well as interactions between the ER and other nuclear proteins. We have examined the potential of the ERalpha to regulate the expression of reporter genes under the control of oestrogen response elements (EREs), NFkappaB response elements (NREs) or AP-1/TPA response elements (TREs) in HeLa cells and in human embryonic kidney (HEK-293) cells. Transiently transfected ERalpha was able to activate expression of beta-galactosidase under the control of EREs in an oestradiol (E2)-dependent manner in both HeLa and HEK-293 cells. The ERalpha was able to repress by 80% the TNF-mediated expression of beta-galactosidase under the control of NREs in an E2-dependent manner in HeLa cells but not in HEK-293 cells. ERalpha/E2 also induced a two-fold potentiation of TPA-mediated expression of beta-galactosidase under the control of TREs in HeLa cells but not in HEK-293 cells. These results suggest that the ERalpha is capable of regulating gene expression in a cell-specific manner. We further investigated the mechanisms by which the ERalpha regulates gene expression in these systems by co-expressing the ERalpha and the reporter gene constructs with known cofactors of the ERalpha. We have shown that expression of steroid receptor coactivator-1 alpha (SRC-1alpha) and receptor interacting protein-140 (RIP-140) have no effect on the capacity of the ERalpha to modulate NFkappaB reporter gene activity in HeLa cells. Furthermore, the expression of SRC-1alpha or RIP-140 does not enable the ERalpha to repress NFkappaB or to potentiate an AP-1 response in HEK-293 cells. This suggests that factors other than SRC-1alpha or RIP-140 are responsible for the cell-specific effects seen with ERalpha.


The anti-progestin RU-486 inhibits viability of MCF-7 breast cancer cells by suppressing WNT1.
The Wnt signaling pathway is activated in over 50% of women with breast cancer and contributes to tumor progression. Here, we investigated the effects of RU-486 on Wnt signaling in breast cancer cell lines. RU-486 reduced viability of the progesterone receptor-positive MCF-7 and T-47D cells, but had no effect on the triple-negative MDA-MB-231 cells. Furthermore, RU-486 suppressed WNT1 expression of MCF-7 cells by 99%. The addition of recombinant WNT1 partially reversed the RU-486-dependent inhibition of viability in MCF-7, but not in T-47D cells. In conclusion, we identified WNT1 as a novel mediator of the anti-tumor effects of RU-486 in MCF-7 cells.


13C-methacetin-breath test compared to also noninvasive biochemical blood tests in predicting hepatic fibrosis and cirrhosis in chronic hepatitis C.
The (13)C-methacetin-breath test and also several noninvasive blood tests comprising routine laboratory parameters have been proposed to predict fibrosis and cirrhosis in chronic hepatitis C. The aim of the study was to compare the diagnostic accuracy between these tests referring to hepatic histology as gold standard. 96 patients with chronic hepatitis C virus infection underwent percutaneous liver biopsy and the (13)C-methacetin-breath test. The Fibroindex, the aspartate aminotransferase to platelet ratio index , and the aspartate aminotransferase to alanine aminotransferase ratio were used as parameters for the staging of fibrosis. The main endpoint was the area under the characteristic curves for the diagnosis of advanced fibrosis (F3-F4) and cirrhosis (F4) according to the Batts Ludwig criteria. ROC analysis revealed a cut-off <14.6 per thousand best with 92.6% sensitivity and 84.1% specificity for the (13)C-methacetin-breath test, for the Fibroindex >1.82 70.4% sensitivity and 91.3% specificity, for the aspartate aminotransferase to platelet ratio >1.0 a 66.7% sensitivity and 75.4% specificity, and for the aspartate aminotransferase to alanine aminotransferase ratio >1.0 63.0% sensitivity and 59.4% specificity in predicting liver cirrhosis. The areas under the curve for the breath test, the Fibroindex, aspartate aminotransferase to platelet ratio and the aspartate aminotransferase to alanine aminotransferase ratio were 0.958, 0.845, 0.799, and 0.688, respectively, when predicting cirrhosis. For identifying patients with advanced fibrosis, the areas under the curve were 0.827, 0.804, 0.779, and 0.561, respectively. Discordances between Fibroindex (21%), aspartate aminotransferase to platelet ratio (29%) or aspartate aminotransferase to alanine aminotransferase ratio (37.6%) and liver biopsy were significantly more frequent than between (13)C-breath test (11.6%) and liver biopsy (P<0.05). The (13)C-methacetin-breath test is more reliable in predicting advanced fibrosis and cirrhosis than simple biochemical parameters (aspartate aminotransferase to platelet ratio; aspartate aminotransferase to alanine aminotransferase ratio).


Incidence of Non-melanoma Eyelid Malignancies in Bulgaria (2000-2015).
Malignant eyelid tumours are the most common malignancies in ophthalmic practice. We report here the incidence of non-melanoma eyelid malignancies in Bulgaria, for the period 2000-2015. All cases coded with the International Classification of Diseases (ICD) code C 44.1, received from the Bulgarian National Cancer Registry, were used for estimating crude incidence rates by age and sex. In addition, directly age standardized incidence rates for non-melanoma eyelid malignancies are presented based on the European Standard population (year 2013) for Bulgaria and the region of Varna. Over the 16-year period there were 3,046 cases in total, of which 1,457 (47.8%) were among men. The age standardized rate (ASR) of non-melanoma eyelid malignancies in men was 2.9 per 100,000 (95% CI 2.79-3.11), in women it was 2.4 per 100,000 (95% CI 2.30-2.54), and the mean ASR was 2.6 per 100,000 (95% CI 2.53-2.73) for both sexes during the period. The ASR increased for women and for both sexes combined.The risk of eyelid malignancies was higher for men than for women with RR of 1.21 (95% CI 1.08-1.41) during the whole period. The risk was higher in women up to the age of 55 and over 55 years it was higher in men. We report results for the incidence of eyelid non-melanoma malignancies in a south-eastern European country, Bulgaria. They are comparable with earlier reports from other European countries.


IRF4 expression is low in Philadelphia negative myeloproliferative neoplasms and is associated with a worse prognosis.
Interferon regulatory factor 4 (IRF4) is involved in the pathogenesis of various hematologic malignancies. Its expression has been related to the negative regulation of myeloid-derived suppressor cells (MDSCs) and the polarization of anti-inflammatory M2 macrophages, thereby altering immunosurveillance and inflammatory mechanisms. An abnormal inflammatory status in the bone marrow microenvironment of myeloproliferative neoplasms (MPNs) has recently been demonstrated; moreover, in chronic myeloid leukemia a downregulated expression of IRF4 has been found. In this context, we evaluated the IRF4 expression in 119 newly diagnosed consecutive Philadelphia negative MPNs (Ph- MPNs), showing a low expression among the MPNs phenotypes with a more significant decrease in primary myelofibrosis patients. Lower IRF4 levels were associated with JAK2 + and triple negatives cases carrying the worst prognosis. Furthermore, the IRF4 levels were related to leukemic transformation and a shorter leukemia-free survival; moreover, the risk of myelofibrosis transformation in polycythemia vera and essential thrombocythemia patients was more frequent in cases with lower IRF4 levels. Overall, our study demonstrates an IRF4 dysregulated expression in MPNs patients and its association with a worse prognosis. Further studies could validate these data, to improve our knowledge of the MPNs pathogenesis and confirm the IRF4 role as a new prognostic factor.


ToxigenicFusarium species isolated from rotted potato tubers.
Fusariumsambucinum(F. trichothecioides),F. avenaceum andF. crookwellense, were the species most frequently isolated from lots of potato tubers imported to Southern Italy in 1985. Isolates of these species were tested for the production of zearalenone, zearalenols, trichothecenes and moniliformin, on three solid media (autoclaved kernels of maize or rice, and fresh potato tubers), incubated at different conditions. The toxigenic potential of the isolates was expressed only on rice and maize, while they were non capable to produce toxins on fresh potato under any conditions. In particular,F. crookwellense produced zearalenone and/or zearalenols (diastereomeric mixture) on maize, andF. avenaceum produced moniliformin both on maize and on rice; while no toxins were produced byF. sambucinum.


Topical application of the pharmacological cold mimetic menthol stimulates brown adipose tissue thermogenesis through a TRPM8, UCP1, and norepinephrine dependent mechanism in mice housed at thermoneutrality.
Increasing whole-body energy expenditure via the pharmacological activation of uncoupling protein 1 (UCP1)-dependent brown adipose tissue (BAT) thermogenesis is a promising weight management strategy, yet most therapeutics studied in rodents to date either induce compensatory increases in energy intake, have thermogenic effects that are confounded by sub-thermoneutral housing temperatures or are not well tolerated in humans. Here, we sought to determine whether the non-invasive topical application of the pharmacological cold mimetic and transient receptor potential (TRP) cation channel subfamily M member 8 (TRPM8) agonist L-menthol (MNTH), could be used to stimulate BAT thermogenesis and attenuate weight gain in mice housed at thermoneutrality. Using three different strains of mice and multiple complimentary approaches to quantify thermogenesis in vivo, coupled with ex vivo models to quantify direct thermogenic effects, we were able to convincingly demonstrate the following: (1) acute topical MNTH application induces BAT thermogenesis in a TRPM8- and UCP1-dependent manner; (2) MNTH-induced BAT thermogenesis is sufficient to attenuate weight gain over time without affecting energy intake in lean and obese mice; (3) the ability of topical MNTH application to stimulate BAT thermogenesis is mediated, in part, by a central mechanism involving the release of norepinephrine. These data collectively suggest that topical application of MNTH may be a promising weight management strategy.


Antimicrobial properties and biocompatibility of electrospun poly-ε-caprolactone fibrous mats containing Gymnema sylvestre leaf extract.
Wound care management presents one of the substantial and tenacious challenges to the healthcare systems worldwide. Microbial colonization and subsequent biofilm formation after injury have garnered much attention, as there is an appreciable correlation between biofilms formation and delayed healing in chronic wounds. Nanotechnology has emerged as a potential platform for the management of treating acute and chronic wounds. This study presents the utility of electrospun nanofiber mats containing a natural extract (Gymnema sylvestre) that averts biofilm formation but supports human dermal fibroblasts (hDFs) attachment. The scaffolds exhibited good wettability, enhanced mechanical properties and contact mediated inhibition of Gram-positive and Gram-negative bacteria. MTS viability assay and confocal imaging further confirmed that the natural extract loaded mats remained non-cytotoxic for hDFs. Overall these findings evidenced the suitability of the Gymnema sylvestre (GS) functionalized electrospun poly-ε-caprolactone (PCL) nanofibers, as an effective wound dressing with broad spectrum anti-bacterial properties.


Metabolic activation of bladder procarcinogens, 2-aminofluorene, 4-aminobiphenyl, and benzidine by Pseudomonas aeruginosa and other human endogenous bacteria.
Pseudomonas aeruginosa, an opportunistic pathogen of the human urinary tract, and other selected human endogenous bacteria were investigated for metabolic activation of the bladder procarcinogens, 2-aminofluorene (2-AF), 4-aminobiphenyl (4-AB), and benzidine (Bz). The cell-free extracts of Pseudomonas aeruginosa, Escherichia coli, Enterobacter aerogenes, Proteus mirabilis, Proteus vulgaris, Staphylococcus epidermidis, Staphylococcus saprophyticus, Klebsiella pneumoniae, and intestinal anaerobes, Bacteroides fragilis, Clostridium perfringens, and Eubacterium aerofaciens produced increased histidine revertant frequencies with the tester strain Salmonella typhimurium TA98 in the Ames Salmonella mutagenicity assay. In addition, the cell-free extracts of Pseudomonas aeruginosa, Bacteroides fragilis, and Eubacterium aerofaciens each showed the presence of a cytochrome P450 absorption peak in the carbon monoxide (CO) difference spectrum. This was not demonstratable for the other bacteria. Our findings indicate that human endogenous bacteria, which are opportunistic pathogens of the urinary bladder, can metabolically activate the bladder procarcinogens 2-AF, 4-AB, and Bz into mutagens. The metabolic activation by Pseudomonas aeruginosa, Bacteroides fragilis, and Eubacterium aerofaciens is mediated by a cytochrome P450 enzyme. For those organisms that induced metabolic activation but did not show a P450 absorption peak with the cell-free extracts, other oxidative enzymes may be involved.