Acetate, a Short-Chain Fatty Acid, Acutely Lowers Heart Rate and Cardiac Contractility Along with Blood Pressure.
Short-chain fatty acids (SCFAs) are metabolites produced almost exclusively by the gut microbiota and are an essential mechanism by which gut microbes influence host physiology. Given that SCFAs induce vasodilation, we hypothesized that they might have additional cardiovascular effects. In this study, novel mechanisms of SCFA action were uncovered by examining the acute effects of SCFAs on cardiovascular physiology in vivo and ex vivo. Acute delivery of SCFAs in conscious radiotelemetry-implanted mice results in a simultaneous decrease in both mean arterial pressure and heart rate (HR). Inhibition of sympathetic tone by the selective β-1 adrenergic receptor antagonist atenolol blocks the acute drop in HR seen with acetate administration, yet the decrease in mean arterial pressure persists. Treatment with tyramine, an indirect sympathomimetic, also blocks the acetate-induced acute drop in HR. Langendorff preparations show that acetate lowers HR only after long-term exposure and at a smaller magnitude than seen in vivo. Pressure-volume loops after acetate injection show a decrease in load-independent measures of cardiac contractility. Isolated trabecular muscle preparations also show a reduction in force generation upon SCFA treatment, though only at supraphysiological concentrations. These experiments demonstrate a direct cardiac component of the SCFA cardiovascular response. These data show that acetate affects blood pressure and cardiac function through parallel mechanisms and establish a role for SCFAs in modulating sympathetic tone and cardiac contractility, further advancing our understanding of the role of SCFAs in blood pressure regulation. SIGNIFICANCE STATEMENT: Acetate, a short-chain fatty acid, acutely lowers heart rate (HR) as well as mean arterial pressure in vivo in radiotelemetry-implanted mice. Acetate is acting in a sympatholytic manner on HR and exerts negative inotropic effects in vivo. This work has implications for potential short-chain fatty acid therapeutics as well as gut dysbiosis-related disease states.


The challenge of using the rheumatoid arthritis diagnostic criteria in clinical practice.
The new 2010 ACR/EULAR (American College of Rheumatology/European League Against Rheumatism) criteria of Rheumatoid Arthritis recently published, have been released to classify and identify patients with early RA who could benefit from early therapy. They recommend anti-citrullinated protein antibody (ACPA) testing as an alternative criterion to Rheumatoid Factor (RF) and ACPA that were introduced together with the other classic criteria in a scoring system. We previously criticized these new criteria because of unavailable specificity and sensibility in the first paper, and the use of ACPA as dichotomous criterion (presence/absent) and alternatives to rheumatoid factor. Our previous work promoted discussion and fostered new research on this issue. By the light of new data, in an effort to improve clinical reasoning, we suggest a more practical probabilistic point of view. In this regard, we analyze the sensitivity and specificity of the diagnostic studies that evaluate the performance of the 2010 classification criteria. Then, we compare the old and the new classification criteria. Subsequently, we describe the use of likelihood ratios applied to the classification criteria and different cutoff levels of ACPA for decision-making in different setting. Moreover, we define some properties of likelihood ratios and their use for diagnosing or excluding rheumatoid arthritis. We want to share this kind of knowledge within the scientific community because we believe that it can help general practitioners and specialists to recognize early arthritis patients implementing a more efficient probabilistic clinical reasoning.


New combinations in the treatment of lung cancer: a time for optimism.
Strides have been made in the treatment of lung cancer in the last decade that warrant a more optimistic outlook toward the disease. The recent development of several new agents with single-agent activity, including paclitaxel, docetaxel, vinorelbine, gemcitabine, and irinotecan, is important, and those agents offer even greater potential when they are used in combination chemotherapy regimens or in combined-modality programs. The experience to date with therapy results with these agents in the treatment of lung cancer is reviewed and is compared to results documented with the current standard treatments for lung cancer, namely, cisplatin and cisplatin-based combination regimens. Published and ongoing trials are outlined, and directions for future research and the future goals of lung cancer therapy are outlined. The availability of newer chemotherapeutic agents that are active in lung cancer has led to response rates as high as 40% in the treatment of non-small cell lung cancer. These drugs have been shown to be active in combination drug regimens as well as when combined with radiotherapy. Future research will focus on using these agents in two- and three-drug regimens as radiation sensitizers and in combination programs with new drugs and biological agents with apparent activity against this disease.


First Predictions of the Angular Power Spectrum of the Astrophysical Gravitational Wave Background.
We present the first predictions for the angular power spectrum of the astrophysical gravitational wave background constituted of the radiation emitted by all resolved and unresolved astrophysical sources. Its shape and amplitude depend on both the astrophysical properties on galactic scales and on cosmological properties. We show that the angular power spectrum behaves as C_{ℓ}∝1/ℓ on large scales and that relative fluctuations of the signal are of order 30% at 100 Hz. We also present the correlations of the astrophysical gravitational wave background with weak lensing and galaxy distribution. These numerical results pave the way to the study of a new observable at the crossroad between general relativity, astrophysics, and cosmology.


Knowledge and use of emergency contraception: a multicountry analysis.
Globally, evidence on knowledge and use of emergency contraception from population-based data is limited, though such information would be helpful in increasing access to the method. We examined knowledge and use of emergency contraception in 45 countries using population-based survey data. Demographic and Health Survey (DHS) data on women aged 15-49 were analyzed by country in logistic regressions to identify associations between women's characteristics and their having heard of emergency contraception or having ever used it. Trends were examined, by region and globally, according to individual, household and community descriptors, including women's age, education, marital status, socioeconomic status, and urban or rural location. The proportion of women who had heard of emergency contraception ranged from 2% in Chad to 66% in Colombia, and the proportion of sexually experienced women who had used it ranged from less than 0.1% in Chad to 12% in Colombia. The odds of having heard of or used the method generally increased with wealth, and although the relationship between marital status and knowing of the method varied by region, never-married women were more likely than married women to have used emergency contraception in countries where significant differences existed. In some countries, urban residence was associated with having heard of the method, but in only three countries were women from urban areas more likely to have used it. Our findings support the need for broader dissemination of information on emergency contraception, particularly among low-income individuals. Variations in use and knowledge within regions suggest a need for programs to be tailored to country-level characteristics.


Arterial elastin as seen with scanning electron microscopy: a review.
All large arteries contain elastin, collagen, and muscle which can be seen with light microscopy and transmission electron microscopy. Elastin forms an internal elastic lamina (IEL) in all arteries, but also forms multiple fenestrated sheets in the media of the aorta and other large arteries. The fenestrations in the media are larger than those in the IEL. The adventitial elastin is more fibrous and often contains tubular elastin surrounding vasa vasorum when prepared by removing all non-elastin by placing the aorta in 0.1 N NaOH at 70-75 degrees C for five hours. The fenestrations are larger near branches and in an experimentally created poststenotic dilatation. Atherosclerosis appears associated with both new elastin formation in early atherosclerosis and elastolysis in late disease.


Analysis of Whole-Genome Sequences for the Prediction of Penicillin Resistance and β-Lactamase Activity in Bacillus anthracis.
Penicillin (PEN) is a low-cost option for anthrax treatment, but naturally occurring resistance has been reported. β-Lactamase expression (bla1, bla2) in Bacillus anthracis is regulated by a sigma factor (SigP) and its cognate anti-sigma factor (RsiP). Mutations leading to truncation of RsiP were previously described as a basis for PEN resistance. Here, we analyze whole-genome sequencing (WGS) data and compare the chromosomal sigP-bla1 regions from 374 B. anthracis strains to determine the frequency of mutations, identify mutations associated with PEN resistance, and evaluate the usefulness of WGS for predicting PEN resistance. Few (3.5%) strains contained at least 1 of 11 different mutations in sigP, rsiP, or bla1. Nine of these mutations have not been previously associated with PEN resistance. Four strains showed PEN resistance (PEN-R) by conventional broth microdilution, including 1 strain with a novel frameshift in rsiP. One strain that carries the same rsiP frameshift mutation as that found previously in a PEN-R strain showed a PEN-susceptible (PEN-S) phenotype and exhibited decreased bla1 and bla2 transcription. An unexpectedly small colony size, a reduced growth rate, and undetectable β-lactamase activity levels (culture supernatant and cell lysate) were observed in this PEN-S strain. Sequence analysis revealed mutations in genes associated with growth defects that may contribute to this phenotype. While B. anthracis rsiP mutations cannot be exclusively used to predict resistance, four of the five strains with rsiP mutations were PEN-R. Therefore, the B. anthracis sigP-bla1 region is a useful locus for WGS-based PEN resistance prediction, but phenotypic testing remains essential. IMPORTANCE Determination of antimicrobial susceptibility of B. anthracis is essential for the appropriate distribution of antimicrobial agents for postexposure prophylaxis (PEP) and treatment of anthrax. Analysis of WGS data allows for the rapid detection of mutations in antimicrobial resistance (AMR) genes in an isolate, but the presence of a mutation in an AMR gene does not always accurately predict resistance. As mutations in the anti-sigma factor RsiP have been previously associated with high-level penicillin resistance in a limited number of strains, we investigated WGS assemblies from 374 strains to determine the frequency of mutations and performed functional antimicrobial susceptibility testing. Of the five strains that contained mutations in rsiP, only four were PEN-R by functional antimicrobial susceptibility testing. We conclude that while sequence analysis of this region is useful for AMR prediction in B. anthracis, genetic analysis should not be used exclusively and phenotypic susceptibility testing remains essential.


Inhibition of microRNA‑124‑3p protects against acute myocardial infarction by suppressing the apoptosis of cardiomyocytes.
The aims of the present study were to investigate the roles and underlying mechanisms of microRNA‑124‑3p (miR‑124‑3p) in the progression of acute myocardial infarction (AMI). The expression of miR‑124‑3p was determined via reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). TargetScan analysis and a luciferase reporter assay were conducted to reveal the association between miR‑124‑3p and nuclear factor κ‑light‑chain‑enhancer of activated B cells (NF‑κB) repressing factor (NKRF). To investigate the role of miR‑124‑3p in AMI, a cell model of myocardial hypoxic/ischemic injury was established by subjecting H9c2 cardiac cells to hypoxia for 48 h. The viability of cells was determined using an MTT assay, and cell apoptosis was analyzed by flow cytometry. Additionally, the expression levels of inflammatory factors [tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β and IL‑6] were measured via ELISA. Furthermore, gene and protein expression levels were determined by performing RT‑qPCR and western blot analyses, respectively. It was revealed that the expression of miR‑124‑3p was significantly increased in the blood of patients with AMI and hypoxia‑treated H9c2 cells. Additionally, it was demonstrated that NKRF was a direct target of miR‑124‑3p. The hypoxia‑induced decrease in the viability of H9c2 cells and increase in cell apoptosis were eliminated by the downregulation of miR‑124‑3p. Furthermore, hypoxia significantly increased the levels of TNF‑α, IL‑1β and IL‑6, whereas miR‑124‑3p downregulation eliminated these effects. Downregulated expression of B‑cell lymphoma 2, pro‑caspase 3 and pro‑caspase 9 protein, and upregulated expression of cleaved caspases 3 and 9 was observed in hypoxic H9c2 cells; the altered expression of these proteins was suppressed by miR‑124‑3p inhibitor. Additionally, miR‑124‑3p inhibitor suppressed the hypoxia‑induced activation of the NF‑κB signaling pathway in H9c2 cells. Furthermore, it was demonstrated that the various effects of miR‑124‑3p inhibitor on H9c2 cells were eliminated by the small interfering RNA‑mediated downregulation of NKRF. In conclusion, the results of the present study indicated that miR‑124‑3p downregulation protected against AMI via inhibition of inflammatory responses and the apoptosis of cardiomyocytes by regulating the NKRF/NF‑κB pathway.


Changes in Underlying Determinants Explain Rapid Increases in Child Linear Growth in Alive & Thrive Study Areas between 2010 and 2014 in Bangladesh and Vietnam.
Background: Child linear growth sometimes improves in both intervention and comparison groups in evaluations of nutrition interventions, possibly because of spillover intervention effects to nonintervention areas or improvements in underlying determinants of nutritional change in both areas.Objective: We aimed to understand what changes in underlying socioeconomic characteristics and behavioral factors are important in explaining improvements in child linear growth.Methods: Baseline (2010) and endline (2014) surveys from the Alive & Thrive impact evaluation were used to identify the underlying determinants of height-for-age z scores (HAZs) among children aged 24-48 mo in Bangladesh (n = 4311) and 24-59 mo in Vietnam (n = 4002). Oaxaca-Blinder regression decompositions were used to examine which underlying determinants contributed to HAZ changes over time.Results: HAZs improved significantly between 2010 and 2014 in Bangladesh (∼0.18 SDs) and Vietnam (0.25 SDs). Underlying determinants improved substantially over time and were larger in Vietnam than in Bangladesh. Multiple regression models revealed significant associations between changes in HAZs and socioeconomic status (SES), food security, maternal education, hygiene, and birth weight in both countries. Changes in HAZs were significantly associated with maternal nutrition knowledge and child dietary diversity in Bangladesh, and with prenatal visits in Vietnam. Improvements in maternal nutrition knowledge in Bangladesh accounted for 20% of the total HAZ change, followed by maternal education (13%), SES (12%), hygiene (10%), and food security (9%). HAZ improvements in Vietnam were accounted for by changes in SES (26%), prenatal visits (25%), hygiene (19%), child birth weight (10%), and maternal education (7%). The decomposition models in both countries performed well, explaining >75% of the HAZ changes.Conclusions: Decomposition is a useful and simple technique for analyzing nonintervention drivers of nutritional change in intervention and comparison areas. Improvements in underlying determinants explained rapid improvements in HAZs between 2010 and 2014 in Bangladesh and Vietnam.


High-definition fiber tracking guidance for intraparenchymal endoscopic port surgery.
The authors have applied high-definition fiber tracking (HDFT) to the resection of an intraparenchymal dermoid cyst by using a minimally invasive endoscopic port. The lesion was located within the mesial frontal lobe, septal area, hypothalamus, and suprasellar recess. Using high-dimensional (256 directions) diffusion imaging, more than 250,000 fiber tracts were imaged before and after surgery. Trajectory planning using HDFT in a computer model was used to facilitate cannulation of the cyst with the endoscopic port. Analysis of the proposed initial surgical route was overlaid onto the fiber tracts and was predicted to produce substantial disruption to prefrontal projection fibers (anterior limb of the internal capsule) and the cingulum. Adjustment of the cannulation entry point 1 cm medially was predicted to cross the corpus callosum instead of the anterior limb of the internal capsule or the cingulum. Following cyst resection performed using endoscopic port surgery, postoperative imaging demonstrated accurate cannulation of the lesion, with improved quantitative signal from both the anterior limb of the internal capsule and the cingulum. The observed fiber preservation from the cingulum and the anterior limb of the internal capsule, with minor injury to the corpus callosum, was in close agreement with preoperative trajectory modeling. Comparison of pre- and postoperative HDFT data facilitated quantification of the benefits and costs of the surgical trajectory. Future studies will help to determine whether HDFT combined with endoscopic port surgery facilitates anatomical and functional preservation in such challenging cases.


Beyond Ecosystem Services: Valuing the Invaluable.
The ecosystem services framework (ESF) is advantageous and widely used for itemising and quantifying ways in which humans benefit from natural places. However, it suffers from two important problems: (i) incoherence of definitions and (ii) a narrow approach to valuation, inadequate to represent the full range of human motives for conservation and the diverse interests of different stakeholders. These shortcomings can lead to a range of problems including double-counting, blind spots and unintended consequences. In this opinion article, we propose an ecosystem valuing framework as a broader and more rigorous way to deliver the benefits currently sought from the ESF, without the conceptual problems.


Spontaneous and induced dome formation by two clonal cell populations derived from a human adenocarcinoma cell line, HT29.
The replacement of glucose by galactose in the culture medium resulted in partial structural and functional enterocytic differentiation of HT29 cells. In order to characterize populations of homogeneously differentiated HT29 cells we have selected two clonal cell lines HT29-D4 and HT29-D9 with the following functional and structural characteristics when grown in a galactose-containing medium: the two clonal cell populations were permanently morphologically differentiated as shown by the presence of mature junctional complexes and a well-organized brush border (especially for HT29-D4 cells); HT29-D4 and HT29-D9 cells were able to form domes early in confluency, which indicated a functional state of differentiation; the process of differentiation was fully reversible when glucose was added to the culture medium. The induction of domes was investigated in these two cell populations and we demonstrated for the first time that proteolytic enzymes are potent inducers of dome formation. The architecture of domes either obtained spontaneously or induced by proteolytic enzymes was not maintained in the presence of ouabain (a specific inhibitor of the Na+/K+-ATPase). In conclusion, HT29-D4 and HT29-D9 cells can be maintained permanently in a differentiated state in a glucose-free medium and were able to form domes at confluency. The observation that proteolytic enzymes were able to induce dome formation can help in the comprehension of the mechanism involved in the establishment of the differentiated state.


Cdc48 and cofactors Npl4-Ufd1 are important for G1 progression during heat stress by maintaining cell wall integrity in Saccharomyces cerevisiae.
The ubiquitin-selective chaperone Cdc48, a member of the AAA (ATPase Associated with various cellular Activities) ATPase superfamily, is involved in many processes, including endoplasmic reticulum-associated degradation (ERAD), ubiquitin- and proteasome-mediated protein degradation, and mitosis. Although Cdc48 was originally isolated as a cell cycle mutant in the budding yeast Saccharomyces cerevisiae, its cell cycle functions have not been well appreciated. We found that temperature-sensitive cdc48-3 mutant is largely arrested at mitosis at 37°C, whereas the mutant is also delayed in G1 progression at 38.5°C. Reporter assays show that the promoter activity of G1 cyclin CLN1, but not CLN2, is reduced in cdc48-3 at 38.5°C. The cofactor npl4-1 and ufd1-2 mutants also exhibit G1 delay and reduced CLN1 promoter activity at 38.5°C, suggesting that Npl4-Ufd1 complex mediates the function of Cdc48 at G1. The G1 delay of cdc48-3 at 38.5°C is a consequence of cell wall defect that over-activates Mpk1, a MAPK family member important for cell wall integrity in response to stress conditions including heat shock. cdc48-3 is hypersensitive to cell wall perturbing agents and is synthetic-sick with mutations in the cell wall integrity signaling pathway. Our results suggest that the cell wall defect in cdc48-3 is exacerbated by heat shock, which sustains Mpk1 activity to block G1 progression. Thus, Cdc48-Npl4-Ufd1 is important for the maintenance of cell wall integrity in order for normal cell growth and division.


Microplastics in a freshwater environment receiving treated wastewater effluent.
Small plastic fragments (microplastics or solid particles <5 mm in size or "microbeads" used in personal care products and cosmetics) may ultimately find their way into aquatic environments. We studied the presence of microplastics (particle sizes 53-105 μm and 106-179 μm) spatially and temporally in 3 connected urban lakes being fed by treated wastewater effluent in Lubbock, Texas. These lakes also serve as drainage during storm events. Water samples from drainage playa wetlands within the city were also collected. Our interest was in determining the presence or absence of microplastics in a freshwater environment as well as the source apportionment between personal care products (via wastewater) and discarded plastics (via runoff). Results showed that average concentrations of microplastics in samples collected from lakes ranged from 0.79 ± 0.88 mg/L to 1.56 ± 1.64 mg/L for the 53-105 μm size fraction and from 0.31 ± 0.72 mg/L to 1.25 ± 1.98 mg/L for the 106-179 μm size fraction. For samples collected from playa wetlands, average microplastic concentrations ranged from 0.64 ± 0.92 mg/L to 5.51 ± 9.09 mg/L for the 53-105 μm size fraction and from nondetectable (ND) to 1.79 ± 3.04 mg/L for the 106-179 μm size fraction. Our results (based on comparison of microplastic masses) suggest that urban runoff also contributes microplastics to surface water in addition to the treated wastewater effluent (in this particular case). The present findings may assist in adopting additional monitoring efforts and provide information on the potential contribution of secondary microplastic input into aquatic environments. Integr Environ Assess Manag 2017;13:528-532. © 2017 SETAC.


Anomalous neural circuit function in schizophrenia during a virtual Morris water task.
Previous studies have reported learning and navigation impairments in schizophrenia patients during virtual reality allocentric learning tasks. The neural bases of these deficits have not been explored using functional MRI despite well-explored anatomic characterization of these paradigms in non-human animals. Our objective was to characterize the differential distributed neural circuits involved in virtual Morris water task performance using independent component analysis (ICA) in schizophrenia patients and controls. Additionally, we present behavioral data in order to derive relationships between brain function and performance, and we have included a general linear model-based analysis in order to exemplify the incremental and differential results afforded by ICA. Thirty-four individuals with schizophrenia and twenty-eight healthy controls underwent fMRI scanning during a block design virtual Morris water task using hidden and visible platform conditions. Independent components analysis was used to deconstruct neural contributions to hidden and visible platform conditions for patients and controls. We also examined performance variables, voxel-based morphometry and hippocampal subparcellation, and regional BOLD signal variation. Independent component analysis identified five neural circuits. Mesial temporal lobe regions, including the hippocampus, were consistently task-related across conditions and groups. Frontal, striatal, and parietal circuits were recruited preferentially during the visible condition for patients, while frontal and temporal lobe regions were more saliently recruited by controls during the hidden platform condition. Gray matter concentrations and BOLD signal in hippocampal subregions were associated with task performance in controls but not patients. Patients exhibited impaired performance on the hidden and visible conditions of the task, related to negative symptom severity. While controls showed coupling between neural circuits, regional neuroanatomy, and behavior, patients activated different task-related neural circuits, not associated with appropriate regional neuroanatomy. GLM analysis elucidated several comparable regions, with the exception of the hippocampus. Inefficient allocentric learning and memory in patients may be related to an inability to recruit appropriate task-dependent neural circuits.


Leveraging Machine Learning for Enantioselective Catalysis: From Dream to Reality.
Catalyst optimization for enantioselective transformations has traditionally relied on empirical evaluation of catalyst properties. Although this approach has been successful in the past it is intrinsically limited and inefficient. To address this problem, our laboratory has developed a fully informatics guided workflow to leverage the power of artificial intelligence (AI) and machine learning (ML) to accelerate the discovery and optimization of any class of catalyst for any transformation. This approach is mechanistically agnostic, but also serves as a discovery platform to identify high performing catalysts that can be subsequently investigated with physical organic methods to identify the origins of selectivity.


Difference in brown adipose tissue effector response and associated thermoresponsiveness of ventromedial hypothalamic (VMH) neurons of 21 degrees C vs. 4 degrees C acclimatized rats to scrotal thermal stimulation.
The present study was designed (1) to determine if scrotal thermal stimulation would activate brown adipose tissue (BAT) thermogenesis, indicated by increases in interscapular BAT temperature (TIBAT) of cold acclimatized (CA, kept at 4 degrees C for 4 weeks) and room temperature acclimatized rats (RA, kept at 21 degrees C for 4 weeks) and (2) to compare the thermoresponsiveness of VMH neurons of both groups to scrotal heating and cooling. VMH extracellular activity was recorded in male RA and CA Sprague-Dawley rats when scrotal temperatures (Tsc) were changed between 5-40 degrees C via localized thermode (3 mm2) along with measurements of TscS and TIBATS. The CA-group showed significant increases in TIBATS during scrotal cooling compared to respective TIBATS of the RA-group. The ratio of VMH warm responsive (WRN), cold responsive (CRN) and temperature non-responsive (TNRN) neurons in the CA-group changed compared to that in the RA-group as a greater percentage of CRNs occurred in the CA-group. Also, the thermoresponsiveness of individual VMH CRNs of the CA rats was significantly increased compared to VMH CRNs of the RA-group. The results indicated that localized scrotal cooling of CA-rats (not RA-rats) can activate BAT thermogenesis. Furthermore, VMH CRNs increased their thermoresponsiveness with chronic cold exposure which may be an important neuronal adaptive response for the subsequent enhanced BAT thermogenic effector response seen in that group.


Docetaxel Reverses Pulmonary Vascular Remodeling by Decreasing Autophagy and Resolves Right Ventricular Fibrosis.
Pulmonary arterial hypertension remains a fatal disease despite the availability of approved vasodilators. Since vascular remodeling contributes to increased pulmonary arterial pressure, new agents that reduce the thickness of pulmonary vascular walls have therapeutic potential. Thus, antitumor agents that are capable of killing cells were investigated. Testing of various antitumor drugs identified that docetaxel is a superior drug for killing proliferating pulmonary artery smooth muscle cells compared with other drugs, including gemcitabine, methotrexate, and ifosfamide. The administration of docetaxel to rats with severe pulmonary arterial hypertension reversed pulmonary vascular remodeling and reduced right ventricular pressure. Docetaxel was found to decrease autophagy as monitored by LC3B-II and p62 expression. The small interfering RNA knockdown of Beclin-1 or LC3B potentiated docetaxel-induced cell death, and knocking down p62 inhibited the docetaxel effects. The suppressed autophagic process is due to the ability of docetaxel to decrease Beclin-1 protein expression in a proteasome-dependent manner. Mass spectrometry identified a novel docetaxel-inducible Beclin-1 binding protein, namely, myosin-9. Knocking down myosin-9 inhibited docetaxel-induced cell death. In damaged right ventricles of pulmonary arterial hypertension rats, docetaxel remarkably promoted the resolution of fibrosis and the regeneration of myocardium. Thus, docetaxel is capable of reversing pulmonary vascular remodeling and resolving right ventricle fibrosis and is a promising therapeutic agent for the treatment of pulmonary arterial hypertension and right heart failure.


Escherichia coli O104 in Feedlot Cattle Feces: Prevalence, Isolation and Characterization.
Escherichia coli O104:H4, an hybrid pathotype of Shiga toxigenic and enteroaggregative E. coli, involved in a major foodborne outbreak in Germany in 2011, has not been detected in cattle feces. Serogroup O104 with H type other than H4 has been reported to cause human illnesses, but their prevalence and characteristics in cattle have not been reported. Our objectives were to determine the prevalence of E. coli O104 in feces of feedlot cattle, by culture and PCR detection methods, and characterize the isolated strains. Rectal fecal samples from a total of 757 cattle originating from 29 feedlots were collected at a Midwest commercial slaughter plant. Fecal samples, enriched in E. coli broth, were subjected to culture and PCR methods of detection. The culture method involved immunomagnetic separation with O104-specific beads and plating on a selective chromogenic medium, followed by serogroup confirmation of pooled colonies by PCR. If pooled colonies were positive for the wzxO104 gene, then colonies were tested individually to identify wzxO104-positive serogroup and associated genes of the hybrid strains. Extracted DNA from feces were also tested by a multiplex PCR to detect wzxO104-positive serogroup and associated major genes of the O104 hybrid pathotype. Because wzxO104 has been shown to be present in E. coli O8/O9/O9a, wzxO104-positive isolates and extracted DNA from fecal samples were also tested by a PCR targeting wbdDO8/O9/O9a, a gene specific for E. coli O8/O9/O9a serogroups. Model-adjusted prevalence estimates of E. coli O104 (positive for wzxO104 and negative for wbdDO8/O9/O9a) at the feedlot level were 5.7% and 21.2%, and at the sample level were 0.5% and 25.9% by culture and PCR, respectively. The McNemar's test indicated that there was a significant difference (P < 0.01) between the proportions of samples that tested positive for wzxO104 and samples that were positive for wzxO104, but negative for wbdDO8/O9/O9a by PCR and culture methods. A total of 143 isolates, positive for the wzxO104, were obtained in pure culture from 146 positive fecal samples. Ninety-two of the 143 isolates (64.3%) also tested positive for the wbdDO8/O9/O9a, indicating that only 51 (35.7%) isolates truly belonged to the O104 serogroup (positive for wzxO104 and negative for wbdDO8/O9/O9a). All 51 isolates tested negative for eae, and 16 tested positive for stx1 gene of the subtype 1c. Thirteen of the 16 stx1-positive O104 isolates were from one feedlot. The predominant serotype was O104:H7. Pulsed-field gel electrophoresis analysis indicated that stx1-positive O104:H7 isolates had 62.4% homology to the German outbreak strain and 67.9% to 77.5% homology to human diarrheagenic O104:H7 strains. The 13 isolates obtained from the same feedlot were of the same PFGE subtype with 100% Dice similarity. Although cattle do not harbor the O104:H4 pathotype, they do harbor and shed Shiga toxigenic O104 in the feces and the predominant serotype was O104:H7.


Cyclic mechanical strain increases production of regulators of bone healing in cultured murine osteoblasts.
The adaptive response of bone to mechanical strain, for which angiogenesis is required, is underscored during fracture healing. Vascular endothelial growth factor (VEGF) and transforming growth factor beta-1 (TGF-beta1) are critical regulators of angiogenesis. The purpose of this study was to examine the effect of strain on the production of VEGF and TGF-beta1. MC3T3-E1 mouse osteoblasts underwent cyclic strain (low, 0.1 Hz, or high, 0.2 Hz) for 24 or 48 hours. VEGF and TGF-beta1 protein levels were determined by ELISA, and Northern blot analysis was performed for VEGF mRNA. Alkaline phosphatase (an osteoblast differentiation marker) activity was determined by functional enzymatic assay. All measurements were standardized for cell number by crystal violet colorimetric assay. Statistical significance was determined by t-test, ANOVA, and the Tukey-Kramer test. Protein production of VEGF and TGF-beta1 was dose-dependently elevated by strain (p < 0.05); alkaline phosphatase did not rise significantly. Northern blot analysis of strained osteoblast cells demonstrated increased VEGF mRNA. Cyclic strain was found to be progressively destructive in a dose-dependent manner, causing 51% and 70% decreases in cell number under low and high strain, respectively (p < 0.01). We demonstrated simultaneous, dose-dependent increases in VEGF and TGF-beta1 protein production by osteoblastic cells in response to increasing strain. VEGF mRNA also increased in response to strain. This strain-induced increase in angiogenic cytokines suggests a potential mechanism by which injured bone may recruit a new blood supply. But we also found increasing strain to increase cellular toxicity, suggesting that cyclic mechanical strain may select for a subpopulation of osteoblasts.


An intramolecular signaling element that modulates dynamin function in vitro and in vivo.
Dynamin exhibits a high basal rate of GTP hydrolysis that is enhanced by self-assembly on a lipid template. Dynamin's GTPase effector domain (GED) is required for this stimulation, though its mechanism of action is poorly understood. Recent structural work has suggested that GED may physically dock with the GTPase domain to exert its stimulatory effects. To examine how these interactions activate dynamin, we engineered a minimal GTPase-GED fusion protein (GG) that reconstitutes dynamin's basal GTPase activity and utilized it to define the structural framework that mediates GED's association with the GTPase domain. Chemical cross-linking of GG and mutagenesis of full-length dynamin establishes that the GTPase-GED interface is comprised of the N- and C-terminal helices of the GTPase domain and the C-terminus of GED. We further show that this interface is essential for structural stability in full-length dynamin. Finally, we identify mutations in this interface that disrupt assembly-stimulated GTP hydrolysis and dynamin-catalyzed membrane fission in vitro and impair the late stages of clathrin-mediated endocytosis in vivo. These data suggest that the components of the GTPase-GED interface act as an intramolecular signaling module, which we term the bundle signaling element, that can modulate dynamin function in vitro and in vivo.


Is prostate cancer different in black men? Answers from 3 natural history models.
Black men in the United States have substantially higher prostate cancer incidence rates than the general population. The extent to which this incidence disparity is because prostate cancer is more prevalent, more aggressive, and/or more frequently diagnosed in black men is unknown. The authors estimated 3 independently developed models of prostate cancer natural history in black men and in the general population using an updated reconstruction of prostate-specific antigen screening, based on the National Health Interview Survey in 2005 and on prostate cancer incidence data from the Surveillance, Epidemiology, and End Results program during 1975 through 2000. By using the estimated models, the natural history of prostate cancer was compared between black men and the general population. The models projected that from 30% to 43% (range across models) of black men develop preclinical prostate cancer by age 85 years, a risk that is (relatively) 28% to 56% higher than that in the general population. Among men who had preclinical disease onset, black men had a similar risk of diagnosis (range, 35%-49%) compared with the general population (32%-44%), but their risk of progression to metastatic disease by the time of diagnosis was from 44% to 75% higher than that in the general population. Prostate cancer incidence patterns implicate higher incidence of preclinical disease and higher risk of metastatic progression among black men. The findings suggest screening black men earlier than white men and support further research into the benefit-harm tradeoffs of more aggressive screening policies for black men. Cancer 2017;123:2312-2319. © 2017 American Cancer Society.


The Role of Zinc and Zinc Homeostasis in Macrophage Function.
Zinc has long been recognized as an essential trace element, playing roles in the growth and development of all living organisms. In recent decades, zinc homeostasis was also found to be important for the innate immune system, especially for maintaining the function of macrophages. It is now generally accepted that dysregulated zinc homeostasis in macrophages causes impaired phagocytosis and an abnormal inflammatory response. However, many questions remain with respect to the mechanisms that underlie these processes, particularly at the cellular and molecular levels. Here, we review our current understanding of the roles that zinc and zinc transporters play in regulating macrophage function.


Finally, a Minimally Invasive Option for Intrahepatic Inferior Vena Cava Invasion by Hepatocellular Carcinoma.
Major vessel invasion is a late manifestation of hepatocellular carcinoma, which may directly result in mortality if left untreated. Surgical resection may be an option for select patients; however, surgery may be contraindicated. Contraindications include multifocal disease, poor liver reserve, inability to tolerate surgery, and patient preference. Cryoablation is a minimally invasive therapy utilized for treating hepatic neoplasms by subjecting them to extreme cold temperatures. The "thermal sink" effect describes cryoablation near flowing blood such as that found in high volumes within major vascular structures. Thermodynamics dictates that proximity to the flowing intravascular compartment will decrease ice formation, and therefore, tumor destruction. This report describes a novel palliative technique to induce temporary cessation of the blood flow in the intrahepatic inferior vena cava (IVC), thus enabling the operator to perform cryoablation targeting invasive cancer within. Adequate ablation using this technique allowed a patient with IVC invasion 9 months of survival. Quality of life during this time was maximized, as recovery time from this procedure is minimal.


Postthrombotic Syndrome in Patients Treated With Rivaroxaban or Warfarin for Venous Thromboembolism.
Postthrombotic syndrome (PTS) is a frequent complication of venous thromboembolism (VTE). Using MarketScan claims data from January 2012 to June 2015, we identified adults with a primary diagnosis code for VTE during a hospitalization/emergency department visit, ≥6 months of insurance coverage prior to the index event and newly started on rivaroxaban or warfarin within 30 days of the index VTE. Patients with <4-month follow-up postindex event or a claim for any anticoagulant during 6-month baseline period were excluded. Differences in baseline characteristics between rivaroxaban and warfarin users were adjusted for using inverse probability of treatment weights based on propensity scores. Patients were followed for the development of PTS starting 3 months after the index VTE. Cox regression was performed and reported as hazard ratios with 95% confidence intervals (CIs). In total, 10 463 rivaroxaban and 26 494 warfarin users were followed for a mean of 16 ± 9 (range, 4-39) months. Duration of anticoagulation was similar between cohorts (median = 6 months). Rivaroxaban was associated with a 23% (95% CI: 16-30) reduced hazard of PTS versus warfarin. Rivaroxaban was associated with a significant risk reduction in symptoms of PTS compared to warfarin in patients with VTE treated in routine practice.


Targeted anti-vascular endothelial growth factor receptor-2 therapy leads to short-term and long-term impairment of vascular function and increase in tumor hypoxia.
Because antiangiogenic therapies inhibit the growth of new tumor-associated blood vessels, as well as prune newly formed vasculature, they would be expected to reduce the supply of oxygen and thus increase tumor hypoxia. However, it is not clear if antiangiogenic treatments lead only to consistent and sustained increases in hypoxia, or transient decreases in tumor hypoxia along with periods of increased hypoxia. We undertook a detailed analysis of an orthotopically transplanted human breast carcinoma (MDA-MB-231) over a 3-week treatment period using DC101, an anti-vascular endothelial growth factor receptor 2 antibody. We observed consistent reductions in microvascular density, blood flow (measured by high-frequency micro-ultrasound), and perfusion. These effects resulted in an increase in the hypoxic tumor fraction, measured with an exogenous marker, pimonidazole, concurrent with an elevation in hypoxia-inducible factor-1alpha expression, an endogenous marker. The increase in tumor hypoxia was evident within 5 days and remained so throughout the entire course of treatment. Vascular perfusion and flow were impaired at days 2, 5, 7, 8, 14, and 21 after the first injection, but not at 4 hours. A modest increase in the vessel maturation index was detected after the 3-week treatment period, but this was not accompanied by an improvement in vascular function. These results suggest that sustained hypoxia and impairment of vascular function can be two consistent consequences of antiangiogenic drug treatment. The implications of the results are discussed, particularly with respect to how they relate to different theories for the counterintuitive chemosensitizing effects of antiangiogenic drugs, even when hypoxia is increased.


Acceptability, effectiveness and cost-effectiveness of blended cognitive-behavioural therapy (bCBT) versus face-to-face CBT (ftfCBT) for anxiety disorders in specialised mental health care: A 15-week randomised controlled trial with 1-year follow-up.
Anxiety disorders are highly prevalent and cause substantial economic burden. Blended cognitive-behavioural therapy (bCBT), which integrates Internet-based CBT and face-to-face CBT (ftfCBT), is an attractive and potentially cost-saving treatment alternative to conventional CBT for patients with anxiety disorders in specialised mental health care. However, little is known about the effectiveness of bCBT in routine care. We examined the acceptability, effectiveness and cost-effectiveness of bCBT versus ftfCBT in outpatient specialised care to patients with panic disorder, social anxiety disorder and generalised anxiety disorder. Patients with anxiety disorders were randomised to bCBT (n = 52) or ftfCBT (n = 62). Acceptability of bCBT and ftfCBT were evaluated by assessing treatment preference, adherence, satisfaction and therapeutic alliance. Costs and effects were assessed at post-treatment and one-year follow-up. Primary outcome measure was the Beck Anxiety Inventory (BAI). Secondary outcomes were depressive symptoms, general psychopathology, work and social adjustment, quality of life and mastery. Incremental cost-effectiveness ratios (ICERs) were computed from societal and healthcare perspectives by calculating the incremental costs per incremental quality-adjusted life year (QALY). No significant differences between bCBT and ftfCBT were found on acceptability or effectiveness measures at post-treatment (Cohen's d between-group effect size on BAI = 0.15, 95% CI -0.30 to 0.60) or at one-year follow-up (d = -0.38, 95% CI -0.84 to 0.09). The modelled point estimates of societal costs (bCBT €10945, ftfCBT €10937) were higher and modelled point estimates of direct medical costs (bCBT €3748, ftfCBT €3841) were lower in bCBT. The acceptability curves showed that bCBT was expected to be a cost-effective intervention. Results should be carefully interpreted due to the small sample size. bCBT appears an acceptable, clinically effective and potentially cost-saving alternative option for treating patients with anxiety disorders. Trials with larger samples are needed to further investigate cost-effectiveness. Netherlands Trial Register: NTR4912.


[Work related predictors for "satisfaction with dementia care" among nurses working in nursing homes].
In German nursing homes dementia care is gaining increasing relevance. Dementia care is known to bear the high risk of a substantial occupational burden among nursing staff. Within this context, the "nurses' satisfaction with the care for residents with dementia" is investigated. Secondary data of the German 3q-study is used to assess degrees of nurses' satisfaction with the care for residents with dementia and potential work related predictors. Data from 813 nurses and nursing aides working in 53 nursing homes were included. 42% of all nursing staff was dissatisfied with the care for residents with dementia in their institution, however, pronounced differences were found between the institutions. Registered nurses and nurses in leading positions were more dissatisfied. A multiple regression analysis indicates that high "quantitative demands", low "leadership quality" and "social interaction with other professions" are strong predictors for nurses' satisfaction with the care for residents with dementia. No association was found for "emotional demands" and "possibilities for development". The results indicate that the "nurses" satisfaction with the care for residents with dementia" may be a highly relevant work factor for nursing staff in nursing homes which deserves additional attention in practice and research. The high predictive power of several work organisational factors implies that preventive action should also include work organisational factors.


Ca(2+)-dependent activator protein for secretion is critical for the fusion of dense-core vesicles with the membrane in calf adrenal chromaffin cells.
Calcium-dependent activator protein for secretion (CAPS) is a neural/endocrine cell-specific protein that has been shown to function at the Ca(2+)-dependent triggering step of dense-core vesicle (DCV) exocytosis in permeabilized PC12 cells. To evaluate the function of CAPS under physiological conditions, we introduced affinity-purified anti-CAPS IgGs into calf adrenal chromaffin (AC) cells via a patch pipette and tested the kinetics of catecholamine secretion using both amperometric and membrane capacitance techniques. The antibodies reacted with a single major approximately 145 kDa protein in AC cells based on immunoblot analysis. AC cells stimulated with sequential trains of action potentials at 7 Hz resulted in successive secretory episodes of equivalent magnitude. When either of two different anti-CAPS IgGs or their Fab fragments were present, a rapid and progressive inhibition of catecholamine release ensued to a maximum of >80%. The effect was specific because preabsorption of IgGs with the respective antigens ablated the inhibitory effect, and the IgGs had no effect on Ca currents. CAPS immunoneutralization not only reduced the number of amperometric spikes but markedly altered the kinetic characteristics of the residual events. The remaining spikes were much smaller (by 85%) and broader (by approximately 3.5-fold) than those in control cells, suggesting that CAPS plays a role in determining release of vesicle contents via the fusion pore. Anti-CAPS IgGs also slowed the rate of the initial exocytotic capacitance burst, representing the docked-and-primed vesicle pool, by approximately 90% but had no effect on the kinetics of rapid endocytosis. These results suggest that CAPS is a key component regulating the fusion of DCVs to the plasma membrane, and possibly fusion pore dilation, in catecholamine secretion from AC cells.


[Liposome fusion with planar lipid membranes].
Potentiodynamic method was used for investigating bilayer lipid membrane (BLM)- liposomes (LS) interaction. BLM was formed from egg lecithin and its mixture with phosphatidylserine (PS); LS-from PS. It is shown that in the presence of calcium in the aqueous phase (1-10 mM) and charged phospholipids in both LS and BLM, fusion of LS with BLM is observed. The fusion event was registered by measuring the increase of negative charge density on BLM surface opposite to that facing the LS.


[Using anti-sera and monoclonal antibodies against Aspergillus fumigatus to study its cross-reactions with Alternaria alteria and Penicillium sp].
Using rabbit polyclonal and mouse monoclonal antibodies against Aspergillus fumigatus, we analyzed their cross-reaction with Alternaria alteria and Penicillium sp. by enzyme immuno assay (EIA) and radioimmunoprecipitation (RIP). It was found that neither of these two categories of antibodies reacted with Alternaria alteria while both reacted with Penicillium sp.. When components from Penicillium sp. precipitated by rabbit antisera were analyzed on 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), a wide range of bands with molecular weights larger than 90 K and a single 70 K band were observed. No band was found after the rabbit antisera was adsorbed with Aspergillus fumigatus. Moreover, two monoclonal antibodies against Aspergillus fumigatus cross-reacted with high molecular weight (greater than 200K) components of Penicillium sp.. These results suggest that the antigen of Alternaria alteria is different from that of Aspergillus fumigatus, and that Penicillium sp. and Aspergillus fumigatus share some antigenic components. In another experiment, human allergic sera containing IgE antibodies to Aspergillus fumigatus were tested against Penicillium sp. by RIP. No cross-reaction was observed. This suggests that in spite of sharing common antigenic components between these two fungi, the allergenic components of Aspergillus fumigatus is different from those of Penicillium sp..


Unique structural properties of DNA interstrand cross-links formed by a new antitumor dinuclear Pt(ii) complex.
The DNA interstrand cross-links of the antitumor drug {[cis-Pt(NH3)2Cl]2(4,4'-methylenedianiline)}2+ (1) play a prevalent role in its antitumor effects. Complex 1 forms DNA long-range interstrand cross-links uniquely in the 3'-3' direction. Conformational distortions induced by these interstrand cross-links in DNA represent a potential structural motif for recognition by high-mobility-group proteins.


Broca and the beginning of modern neurosurgery.
The study presents Broca's pioneering efforts on cerebral localizations and craniotopography and their application on the first craniotomy based on cerebral localization.


Hyaluronic Acid-Modified Cationic Lipid-PLGA Hybrid Nanoparticles as a Nanovaccine Induce Robust Humoral and Cellular Immune Responses.
Here, we investigated the use of hyaluronic acid (HA)-decorated cationic lipid-poly(lactide-co-glycolide) acid (PLGA) hybrid nanoparticles (HA-DOTAP-PLGA NPs) as vaccine delivery vehicles, which were originally developed for the cytosolic delivery of genes. Our results demonstrated that after the NPs uptake by dendritic cells (DCs), some of the antigens that were encapsulated in HA-DOTAP-PLGA NPs escaped to the cytosolic compartment, and whereas some of the antigens remained in the endosomal/lysosomal compartment, where both MHC-I and MHC-II antigen presentation occurred. Moreover, HA-DOTAP-PLGA NPs led to the up-regulation of MHC, costimulatory molecules, and cytokines. In vivo experiments further revealed that more powerful immune responses were induced from mice immunized with HA-DOTAP-PLGA NPs when compared with cationic lipid-PLGA nanoparticles and free ovalbumin (OVA); the responses included antigen-specific CD4(+) and CD8(+) T-cell responses, the production of antigen-specific IgG antibodies and the generation of memory CD4(+) and CD8(+) T cells. Overall, these data demonstrate the high potential of HA-DOTAP-PLGA NPs for use as vaccine delivery vehicles to elevate cellular and humoral immune responses.


Relationship between clinical and pathologic features of ductal carcinoma in situ and patient age: an analysis of 657 patients.
Prior studies have shown that young patient age at diagnosis is associated with an increased risk of local recurrence among women with ductal carcinoma in situ (DCIS) treated with breast-conserving therapy. Whether this can be explained by differences in clinical or pathologic features of DCIS according to age is an unresolved issue. We compared clinical and pathologic features of DCIS among 657 women in 4 age groups: <45 years (n=111), 45 to 54 years (n=191), 55 to 64 years (n=160), and 65+ years (n=195). DCIS presented as a mammographic abnormality less often in younger than in older women (68%, 82%, 81%, and 86% for women <45, 45 to 54, 55 to 64, and 65+ y, respectively; P=0.003). Among the pathologic features analyzed, DCIS extent as determined by the number of low power fields was greater in younger than in older women (mean number of low power fields were 18.6, 14.2, 10.8, and 11.3 in women <45, 45 to 54, 55 to 64 and 65+ y; P<0.001). In addition, cancerization of lobules was present more often in younger than in older women (77%, 73%, 66%, and 50% for women <45, 45 to 54, 55 to 64 and 65+ y, respectively; P<0.0001). Of note, we found no statistically significant relationship between age and DCIS architectural pattern, nuclear grade, comedo necrosis or expression of estrogen receptor, progesterone receptor or human epidermal growth factor receptor 2. We conclude that DCIS in younger women is more often symptomatic, is more extensive, and more often shows cancerization of lobules than DCIS in older women. Whether these features contribute to the higher local recurrence risk in young women with DCIS treated with the breast-conserving therapy requires further study.


A rare diagnosis: Acute bilateral vasitis mimicking incarcerated bilateral inguinal hernia.
Inflammation of the vas deferens or what known as acute vasitis is an under-reported condition that usually presents with scrotal or inguinal pain and swelling which can be misdiagnosed and treated as many other conditions. Here, we present one of the first cases to be reported in the literature with bilateral manifestation. A 28-year-old male patient presented complaining of bilateral testicular and inguinal pain associated with inguinal swelling for 3 days. Initially, the patient was being evaluated as a case of incarcerated bilateral inguinal hernia, but with the aid of radiological imaging, the patient was diagnosed correctly and the unnecessary surgical intervention was prevented.


Gene expression levels are correlated with synonymous codon usage, amino acid composition, and gene architecture in the red flour beetle, Tribolium castaneum.
Gene expression levels correlate with multiple aspects of gene sequence and gene structure in phylogenetically diverse taxa, suggesting an important role of gene expression levels in the evolution of protein-coding genes. Here we present results of a genome-wide study of the influence of gene expression on synonymous codon usage, amino acid composition, and gene structure in the red flour beetle, Tribolium castaneum. Consistent with the action of translational selection, we find that synonymous codon usage bias increases with gene expression. However, the correspondence between tRNA gene copy number and optimal codons is weak. At the amino acid level, translational selection is suggested by the positive correlation between tRNA gene numbers and amino acid usage, which is stronger for highly expressed genes. In addition, there is a clear trend for increased use of metabolically cheaper, less complex amino acids as gene expression increases. tRNA gene numbers also correlate negatively with amino acid size/complexity (S/C) score indicating the coupling between translational selection and selection to minimize the use of large/complex amino acids. Interestingly, the analysis of 10 additional genomes suggests that the correlation between tRNA gene numbers and amino acid S/C score is widespread and might be explained by selection against negative consequences of protein misfolding. At the level of gene structure, three major trends are detected: 1) complete coding region length increases across low and intermediate expression levels but decreases in highly expressed genes; 2) the average intron size shows the opposite trend, first decreasing with expression, followed by a slight increase in highly expressed genes; and 3) intron density remains nearly constant across all expression levels. These changes in gene architecture are only in partial agreement with selection favoring reduced cost of biosynthesis.


Prospects for bronchodilating drugs in the 1990s.
There is likely to be a continuing need for bronchodilator drugs until the time that the primary underlying defect in asthma is determined. At the moment there are several interesting and exciting developments both in the areas of well established drugs (beta agonists, antimuscarinic drugs and phosphodiesterase inhibitors) and in new areas. The potassium channel activators are clearly of interest; more information on their therapeutic ratio is needed. It may be that antagonists or inhibitors of leukotrienes or PAF will find a role and there should be some answers shortly from the studies that are currently underway. Opioid analogues may have a place though at present they must be regarded as rather speculative. Greater understanding of cell signal transduction should provide a further opportunity to modulate smooth muscle contraction and the function of other cells relevant to asthma.


Accounting for teaching hospitals' higher costs and what to do about them.
Academic health centers (AHCs) have higher costs per case and also lower margins than either other teaching hospitals or community hospitals. The differences in margins stem mostly from differences in the intensity with which similar patients are treated, as well as hospitals' ability to generate revenue to cover the costs of that greater intensity, rather than graduate medical education per se. How much patient care capacity should be supported in AHCs and who should be treated with the greater intensity they offer are open questions. If there is to be a public trust fund to subsidize AHCs, it should be financed from general revenues.


Stimulation of NMDA and AMPA receptors in the rat nucleus basalis of Meynert affects sleep.
The nucleus basalis of Meynert (NBM), a heterogeneous area in the basal forebrain involved in the modulation of sleep and wakefulness, is rich in glutamate receptors, and glutamatergic fibers represent an important part of the input to this nucleus. With the use of unilateral infusions in the NBM, the effects of two different glutamatergic subtype agonists, namely N-methyl-D-aspartic acid (NMDA) and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) hydrobromide, on sleep and wakefulness parameters were determined in freely moving rats by means of polygraphic recordings. NMDA (5 nmol) and AMPA (0.4 nmol) induced an increase in wakefulness and an inhibition of slow-wave sleep. AMPA, but not NMDA, also caused a decrease in desynchronized sleep. These AMPA- and NMDA-mediated effects were counteracted by a pretreatment with the specific NMDA antagonist 2-amino-5-phosphonopentanoic acid (20 nmol) and the specific AMPA antagonist 6,7-dinitroquinoxaline-2,3-dione (2 nmol), respectively. The results reported here indicate that 1) the NBM activation of both NMDA and AMPA glutamate receptors exert a modulatory influence on sleep and wakefulness, and 2) AMPA, but not NMDA receptors, are involved in the modulation of desynchronized sleep, suggesting a different role for NBM NMDA and non-NMDA receptors in sleep modulation.


Social Anxiety and Mental Health Service Use Among Asian American High School Students.
Asian American adults endorse more symptoms of social anxiety (SA) on self-report measures than European Americans, but demonstrate lower prevalence rates of SA disorder in epidemiological studies. These divergent results create ambiguity concerning the mental health needs of Asian Americans. The present study is the first to investigate this issue in adolescents through assessment of self-reported SA in Asian American high school students. Parent and self-ratings of impairment related to SA and self-reported mental health service use for SA were also measured. Asian American students endorsed a greater number of SA symptoms and scored in the clinical range more frequently than other ethnic groups. Also, Asian American and Latino students endorsed more school impairment related to SA than other ethnic groups. No differences in parent-reported impairment or service utilization were identified. Implications for future research and treatment for SA among Asian American adolescents are discussed.


Truthfulness Predominates in Americans' Conceptualizations of Honesty: A Prototype Analysis.
Honesty is a near universally valued trait. However, the term honesty captures a litany of traits and behaviors, obscuring research on social perceptions and trait measurement of honesty and creating philosophical difficulties in accounting for what (if anything) unifies this diversity. We applied a prototype analysis approach to identify the most central elements of lay honesty conceptualizations, identifying elements that come to mind and are explicitly acknowledged as important to honesty. In five studies (N = 1,442), U.S. American participants generated 6,000+ free responses characterizing honesty and indicated which subtraits and behaviors best represent honesty. Truthfulness was most central to lay honesty conceptualizations across all studies and several centrality indices (frequency among responses and participants, agreement across participants, priority in lists, explicit ratings), though several other features were prominent. Findings illuminate social perceptions of honesty, critique popular measurement of trait honesty, and offer empirical foundations for philosophical analysis of honesty.


Association of PCSK9 Variants With the Risk of Atherosclerotic Cardiovascular Disease and Variable Responses to PCSK9 Inhibitor Therapy.
Genetic polymorphisms or variations, randomly distributed in a population, may cause drug-gene response variations. Investigation into these polymorphisms may identify novel mechanisms contributing to a specific disease process. Such investigation necessitates the use of Mendelian randomization, an analytical method that uses genetic variants as instrumental variables for modifiable risk factors that affect population health.1 In the past decade, advances in our understanding of genetic polymorphisms have enabled the identification of genetic variants in candidate genes that impact low-density lipoprotein cholesterol (LDL-C) regulating pathways and cardiovascular disease (CVD) outcomes. A specific candidate gene of interest is that of the LDL receptor degrading protein, PCSK9. In fact, loss-of-function genetic variants for the PCSK9 gene are what first highlighted this pathway as a candidate for pharmacologic inhibition. PCSK9 inhibitors (PCSK9i) are a class of cholesterol-lowering medications that provide significant reductions in LDL by inhibiting the degradation of LDL receptors (LDLR). These inhibitors have also been found to reduce production and enhance clearance of lipoprotein A (Lp[a]), an LDL-like particle currently under study as a separate risk factor for atherosclerotic CVD. Here, we discuss the promise of personalized medicine in developing a more efficacious and individualized pharmacogenomics-based approach for the use of PCSK9i that considers genetic variation and targets different patient populations. This review explores the pharmacogenomics of PCSK9i in the context of PCSK9 allele variants related to drug-metabolizing enzymes and responses since more studies are demonstrating that some patients are hyporesponsive or non-responsive to PCSK9i.2 In summary, the pharmacogenomics of PCSK9 are a promising therapeutic target and genetic information from prospective randomized clinical trials is warranted to gain a full understanding of the efficacy and cost-effectiveness of such allele and/or gene-guided PCSK9i therapy.


Cancer risk across mammals.
Cancer is a ubiquitous disease of metazoans, predicted to disproportionately affect larger, long-lived organisms owing to their greater number of cell divisions, and thus increased probability of somatic mutations1,2. While elevated cancer risk with larger body size and/or longevity has been documented within species3-5, Peto's paradox indicates the apparent lack of such an association among taxa6. Yet, unequivocal empirical evidence for Peto's paradox is lacking, stemming from the difficulty of estimating cancer risk in non-model species. Here we build and analyse a database on cancer-related mortality using data on adult zoo mammals (110,148 individuals, 191 species) and map age-controlled cancer mortality to the mammalian tree of life. We demonstrate the universality and high frequency of oncogenic phenomena in mammals and reveal substantial differences in cancer mortality across major mammalian orders. We show that the phylogenetic distribution of cancer mortality is associated with diet, with carnivorous mammals (especially mammal-consuming ones) facing the highest cancer-related mortality. Moreover, we provide unequivocal evidence for the body size and longevity components of Peto's paradox by showing that cancer mortality risk is largely independent of both body mass and adult life expectancy across species. These results highlight the key role of life-history evolution in shaping cancer resistance and provide major advancements in the quest for natural anticancer defences.


Shortage of perioperative drugs: implications for anesthesia practice and patient safety.
Several medications used in clinical perioperative medicine are currently cited on the national shortage list. Medication shortages may be attributed to lack of raw materials, manufacturing issues, and discontinuation of production. Medication shortage has a substantial impact on patient care, and is responsible for creating an environment conducive to an increase in medication errors. Anesthesiologists should be taking an active role with the pharmacy and hospital management to alert caregivers and help to prevent adverse effects on patient care and safety.


Haploid selection, sex ratio bias, and transitions between sex-determining systems.
Sex determination is remarkably dynamic; many taxa display shifts in the location of sex-determining loci or the evolution of entirely new sex-determining systems. Predominant theories for why we observe such transitions generally conclude that novel sex-determining systems are favoured by selection if they equalise the sex ratio or increase linkage with a locus that experiences different selection in males versus females. We use population genetic models to extend these theories in two ways: (1) We consider the dynamics of loci very tightly linked to the ancestral sex-determining loci, e.g., within the nonrecombining region of the ancestral sex chromosomes. Variation at such loci can favour the spread of new sex-determining systems in which the heterogametic sex changes (XY to ZW or ZW to XY) and the new sex-determining region is less closely linked (or even unlinked) to the locus under selection. (2) We consider selection upon haploid genotypes either during gametic competition (e.g., pollen competition) or meiosis (i.e., nonmendelian segregation), which can cause the zygotic sex ratio to become biased. Haploid selection can drive transitions between sex-determining systems without requiring selection to act differently in diploid males versus females. With haploid selection, we find that transitions between male and female heterogamety can evolve so that linkage with the sex-determining locus is either strengthened or weakened. Furthermore, we find that sex ratio biases may increase or decrease with the spread of new sex chromosomes, which implies that transitions between sex-determining systems cannot be simply predicted by selection to equalise the sex ratio. In fact, under many conditions, we find that transitions in sex determination are favoured equally strongly in cases in which the sex ratio bias increases or decreases. Overall, our models predict that sex determination systems should be highly dynamic, particularly when haploid selection is present, consistent with the evolutionary lability of this trait in many taxa.


Importance of literacy for self-reported health-related quality of life: a study of boys with haemophilia in Brazil.
Psychosocial outcomes are important in the perspective of boys with haemophilia. However, health-related quality of life (HRQoL) is based on self-report, and assumes adequate literacy. Yet, literacy is rarely assessed prior to data collection. This study sought to identify criteria that might indicate the level of literacy of children being recruited for clinical trials and to develop a simple method to prescreen those whose literacy was uncertain. We developed a brief screening tool in the form of two stories, at a grade 3 reading level, followed by comprehension questions. We applied the screening test to a sample of haemophilic boys between the ages of 7 and 13 years to assess their literacy. The data were analysed to determine the best criteria to use in identifying the ability to independently self-report for HRQoL studies. Twenty-four Brazilian boys (7.9-12.8) completed the testing. The results showed that 17 (70.8%) were literate (were able to both read and comprehend), and could complete a questionnaire without assistance. All boys over 11.0 years of age were sufficiently literate. Grade level was not found to be a helpful criterion. We recommend that all children under the age of 11.0 years be prescreened before providing self-reported HRQoL data. Those with limited literacy should be provided assistance to ensure comprehension of the questions. This is important to ensure high-quality data on HRQoL for future clinical trials.


Selective antagonism by naltrindole of the antinociceptive effects of the delta opioid agonist cyclic[D-penicillamine2-D-penicillamine5]enkephalin in the rat.
Spinal delta opioid receptors have been proposed to mediate antinociception in the rat on the basis of 1) the efficacy of a small number of agonists; 2) the lack of effect of mu-selective antagonists; and 3) the lack of cross-tolerance with mu-selective agonists. However, direct evidence to support or refute this postulate has not been obtained in the rat due to a lack of suitable delta-selective antagonists. The present study characterized the ability of Naltrindole (NTI, 17-cyclopropylmethyl-6,7-dehydro-4,5 alpha-epoxy-3,14-dihydroxy-6,7-2',3'-indolomorphinan), a recently discovered delta-selective antagonist, to antagonize the antinocieption produced by intrathecal (i.t.) administration of the prototypic delta-selective agonist cyclic[D-penicillamine2-D-penicillamine5]enkephalin (DPDPE) or the mu-selective agonists morphine and [D-Ala2,MePhe4,Gly-ol5] enkephalin (DAMGO) in the rat. Intrathecal coadministration of NTI with DPDPE significantly antagonized the increase in tail-flick latency (TFL) and hot-plate latency (HPL) produced by DPDPE. In the absence of NTI, the ED50 values and 95% CL of DPDPE in the tail-flick and hot-plate tests were 2.8 (1.1-4.7) and 19.5 (13.3-33.7) micrograms, respectively. In the presence of 10 micrograms of NTI, the ED50 value of DPDPE in the tail-flick test was unchanged and was increased by 2-fold in the hot-plate test to 35.9 (26.2-60.1) micrograms. In the presence of 30 micrograms of NTI, the ED50 value of DPDPE in the tail-flick test was increased by 5-fold to 14.5 (8.5-24.9) micrograms and its antinociceptive effect in the hot-plate test was antagonized completely.(ABSTRACT TRUNCATED AT 250 WORDS)


The prognostic contribution of CBL, NRAS, KRAS, RUNX1, and TP53 mutations to mutation-enhanced international prognostic score systems (MIPSS70/plus/plus v2.0) for primary myelofibrosis.
Contemporary risk models in primary myelofibrosis (PMF) include the mutation (MIPSS70) and mutation/karyotype enhanced (MIPSS70 plus/v2.0) international prognostic scoring systems. High molecular risk (HMR) mutations incorporated in one or both of these models include ASXL1, SRSF2, EZH2, IDH1/2, and U2AF1Q157; the current study examines additional prognostic contribution from more recently described HMR mutations, including CBL, NRAS, KRAS, RUNX1, and TP53. In a cohort of 363 informative cases (median age 58 years; 60% males), mutations included JAK2 61%, CALR 24%, MPL 6%, ASXL1 29%, SRSF2 10%, U2AF1Q157 5%, EZH2 10%, IDH1/2 4%, TP53 5%, CBL 5%, NRAS 7%, KRAS 4%, and RUNX1 4%. At a median follow-up of 4.6 years, 135 (37%) deaths and 42 (11.6%) leukemic transformations were recorded. Univariate analysis confirmed significant survival impact from the original MIPSS70/plus/v2.0 HMR mutations as well as CBL (HR 2.8; p < .001), NRAS (HR 2.4; p < .001), KRAS (HR 2.1; p = .01), and TP53 (HR 2.4; p = .004), but not RUNX1 mutations (HR 1.8; p = .08). Multivariate analysis (MVA) that included both the original and more recently described HMR mutations confirmed independent prognostic contribution from ASXL1 (HR 1.8; p = .007), SRSF2 (HR 4.3; p < .001), U2AF1Q157 (HR 2.9, p = .004), and EZH2 (HR 2.4; p < .001), but not from IDH1/2 (p = .3), TP53 (p = .2), CBL (p = .3), NRAS (p = .8) or KRAS (p = .2) mutations. The lack of additional prognostic value from CBL, NRAS, KRAS, RUNX1, and TP53 was further demonstrated in the setting of (i) MVA of mutations and karyotype, (ii) MVA of MIPSS70/plus/v2.0 composite scores and each one of the recently described HMR mutations, except TP53, and iii) modified MIPSS70/plus/plus v2.0 that included CBL, NRAS, KRAS, and TP53 as part of the HMR constituency, operationally referred to as "HMR+" category. Furthermore, "HMR+" enhancement of MIPSS70/plus/plus v2.0 did not result in improved model performance, as measured by C-statistics. We conclude that prognostic integrity of MIPSS70/plus/plus v2.0, as well as their genetic components, was sustained and their value not significantly upgraded by the inclusion of more recently described HMR mutations, including CBL, NRAS, KRAS, and RUNX1. Additional studies are needed to clarify the apparent additional prognostic value of TP53 mutation and its allelic state.


The Knosp Criteria Revisited: 3-Dimensional Volumetric Analysis as a Predictive Tool for Extent of Resection in Complex Endoscopic Pituitary Surgery.
The Knosp criteria have been the historical standard for predicting cavernous sinus invasion, and therefore extent of surgical resection, of pituitary macroadenomas. Few studies have sought to reappraise the utility of this tool after recent advances in visualization and modeling of tumors in complex endoscopic surgery. To evaluate our proposed alternative method, using 3-dimensional (3D) volumetric imaging, and whether it can better predict extent of resection in nonfunctional pituitary adenomas. Patients who underwent endoscopic transsphenoidal resection of pituitary macroadenomas at our institution were reviewed. Information was collected on neurological, endocrine, and visual function. Volumetric segmentation was performed using 3D Slicer software. Relationship of tumor volume, clinical features, and Knosp grade on extent of resection was examined. One hundred forty patients were identified who had transsphenoidal resection of nonfunctional pituitary adenomas. Macroadenomas had a median volume of 6 cm 3 (IQR 3.4-8.7), and 17% had a unilateral Knosp grade of at least 3B. On multiple logistic regression, only smaller log-transformed preoperative tumor volume was independently associated with increased odds of gross total resection (GTR; odds ratio: 0.27, 95% CI: 0.07-0.89, P < .05) when controlling for tumor proliferative status, age, and sex (area under the curve 0.67). The Knosp criteria did not independently predict GTR in this cohort ( P > .05, area under the curve 0.46). Increasing use of volumetric 3D imaging may better anticipate extent of resection compared with the Knosp grade metric and may have a greater positive predictive value for GTR. More research is needed to validate these findings and implement them using automated methods.


Development and Validation of a Nomogram to Predict the Risk of Recurrent Lower Extremity Radiating Pain Within 1 Week Following Full-Endoscopic Lumbar Discectomy.
Accurately predicting the risk of lower extremity (LE) radiating pain after surgery is an important endeavor for spinal surgeons. Our study aimed to identify risk factors for LE radiating pain after decompression with full-endoscopic lumbar discectomy (FELD) and develop a nomogram. We retrospectively reviewed the medical data of patients with lumbar disc herniation who underwent FELD. Two hundred thirty-five patients diagnosed at our hospital from January 2015 to December 2020 were used for model development. The independent risk factors for LE radiating pain after surgery were determined by least absolute shrinkage and selection operator logistic regression and multivariate logistic regression analysis. A nomogram was developed to predict the risk of LE radiating pain based on independent risk factors. Receiver operating characteristic curve, calibration curve, and decision curve analyses were used to evaluate the predictive performance. The nomogram was further verified by an independent cohort. Three hundred seventy-five patients were enrolled in this study, with 102 patients in the training cohort reporting LE radiating pain after FELD, while 133 patients did not. In the validation cohort, 57 patients reported LE radiating pain after FELD, while 83 patients did not. The model was established by multivariate logistic regression analysis. The risk factors included a higher Michigan State University classification of herniated discs, increased disease course, increased time of surgery, reduced lateral recess width, and an interlaminar surgical approach, compared to transforaminal approach. The C-indices and the area under the receiver operating characteristic curve of the predictive model demonstrated good discrimination. Good predictive performance and accuracy were also observed in the validation cohort. A novel nomogram for predicting recurrent LE radiating pain within 1 week after FELD was established and validated. More aggressive pain management strategies should be considered for patients at high risk of LE radiating pain after surgery, as predicted by this model.


Comparative proteome analysis of the strawberry-Fusarium oxysporum f. sp. fragariae pathosystem reveals early activation of defense responses as a crucial determinant of host resistance.
Fusarium wilt on strawberry caused by Fusarium oxysporum f. sp. fragariae (Fof) is a serious threat to commercial strawberry production worldwide. However, resistance mechanisms of strawberry against Fof remain unknown. To reveal the defense responses of strawberry against Fof, comparative proteome analyses were conducted to determine temporal changes in root proteomes of the resistant cv. Festival and susceptible cv. Camarosa from 4 to 72 h post inoculation with Fof. Analysis of proteins separated by two-dimensional gel electrophoresis revealed 79 Fof-responsive proteins with significant differences in abundance (P < 0.05 and greater than 2-fold) in the resistant and/or susceptible cultivar. The 79 proteins were identified through MALDI-TOF/TOF MS/MS analysis, and were mainly involved in primary, secondary and protein metabolism, stress and defense responses, antioxidant and detoxification mechanisms, and hormone biosynthesis. Among these, pathogenesis-related proteins and proteins involved in reactive oxygen species detoxification, ethylene/jasmonic acid signaling pathways, secondary metabolite biosynthesis, glycolysis and/or ubiquitin/26S proteasome-mediated protein degradation have great potential in mediating strawberry resistance against Fof. Protein modification may also have an important contribution. This study provides the first insights into strawberry resistance mechanisms against Fof, opening novel avenues to engineer new strawberry cultivars with improved disease resistance and to develop more effective and sustainable disease management strategies.


Chest Pain During Cesarean Delivery in Relation to Anomalous Right Coronary Artery: A Case Report.
Chest pain during labor can have many etiologies, one of which is myocardial ischemia. We report a 38-year-old woman with American Society of Anesthesiology physical status I scheduled for elective cesarean delivery who experienced chest pain and electrocardiographic signs of myocardial ischemia immediately after carbetocin administration. Subsequent investigations revealed an aberrant right coronary artery originating from the left coronary sinus coursing between the aorta and the pulmonary artery. This was considered the likely cause of the patient's chest pain. The patient gave written permission for the authors to publish this report.


Recurrent vasculitis in cystic fibrosis.
A 22-year-old man presented with recurrent palpable purpuric rash. His clubbing relates to underlying cystic fibrosis (CF) and his rash was identified as CF-related vasculitis, a rare extrapulmonary manifestation of the disease. It occurs predominantly on the lower limbs, mainly over the dorsa of the feet, ankles and tibial surfaces. The rash occurred while the patient had an infective exacerbation of CF (IECF), however, there had also been previous occurrences without worsening of his pulmonary symptoms, to which the rash remitted spontaneously. The patient responded well to immunosuppression, which was given on this admission due to worsening of his CF-related vasculitis. He died 18 months within the onset of his initial rash.


Should we still use Camitta's criteria for severe aplastic anemia?
The criteria by Camitta for diagnosis in severe aplastic anemia (SAA) has been used since 1976. However, there has been no attempt to verify the Camitta's criteria, that the survival in patients with SAA may differ by absolute neutrophil count (ANC), platelet count (PLT), and corrected reticulocyte count (CRC), which are components of the Camitta's criteria. 117 SAA patients diagnosed by the Camitta's criteria were analyzed, retrospectively. Univariate and multivariate analyses were used to evaluate the factors affecting overall survival (OS). Response by immunosuppressive therapy (IST) or stem cell transplantation (SCT) significantly affected OS (P=0.001). Therefore, we excluded treatment responders for analysis. Finally, 92 SAA patients including treatment non-responders by IST or SCT and conservative care group were analyzed by using univariate and multivariate analyses. The median age of analyzed patients was 54.5 years. Male to female ratio was 1:1. The median follow-up duration was 74.23 months (range, 54.71-93.74 months). The median ANC, PLT, and CRC were 394/µL, 12,000/µL, and 0.39%, respectively. In multivariate analyses, ANC <500/µL or ≥500/µL (P=0.015, HR 2.694, 95% CI: 1.20-6.01) and age (P=0.015, HR 1.022, 95% CI: 1.00-1.04) were the significant factors for OS. ANC could be an essential, not an optional criterion for diagnosing SAA. This study suggests the possibility that the Camitta's criteria be modified. Studies in large cohorts are needed to transform the Camitta's criteria.


A liver stiffness-based etiology-independent machine learning algorithm to predict hepatocellular carcinoma.
The existing hepatocellular carcinoma (HCC) risk scores have modest accuracy and most are specific to chronic hepatitis B infection. In this study, we developed and validated a liver stiffness-based machine learning algorithm (ML) for prediction and risk stratification of HCC in various chronic liver diseases (CLDs). MLs were trained for prediction of HCC in 5155 adult patients with various CLDs in Korea and further tested in two prospective cohorts from Hong Kong (HK, N=2732) and Europe (N=2384). Model performance was assessed according to Harrell's C-index and time-dependent receiver operating characteristic (ROC) curve. We developed the SMART-HCC score, a liver stiffness-based ML HCC risk score, with liver stiffness measurement ranked as the most important among 9 clinical features. The Harrell's C-index of the SMART-HCC score in HK and Europe validation cohorts were 0.89 (95% confidence interval [CI] 0.85-0.92) and 0.91 (95%CI 0.87-0.95), respectively. The area under ROC curves of the SMART-HCC score for HCC in 5 years were ≥0.89 in both validation cohorts. The performance of SMART-HCC score was significantly better than existing HCC risk scores including aMAP score, Toronto HCC risk index, and seven hepatitis B related risk scores. Using dual cut-offs of 0.043 and 0.080, the annual HCC incidence was 0.09%-0.11% for low-risk group, and 2.54%-4.64% for high-risk group in the HK and Europe validation cohorts. The SMART-HCC score is a useful machine learning-based tool for clinicians to stratify HCC risk in patients with CLDs.


Retinoid-suppressed phosphorylation of RARalpha mediates the differentiation pathway of osteosarcoma cells.
Although retinoic acid (RA) is a potent agent that coordinates inhibition of proliferation with differentiation of many cell types, RA-mediated signaling pathways in osteosarcoma cell differentiation are uncharacterized. In this study, we show that in human U2OS osteosarcoma cells, decreased phosphorylation of RA receptor alpha (RARalpha) by RA treatment or overexpressing a phosphorylation-defective mutant RARalphaS77A results in the inhibition of proliferation and induction of differentiation, and that U2OS cells transduced with RARalphaS77A suppresses tumor formation in nude mice. Moreover, using different human primary osteosarcoma cells and human mesenchymal stem cells for gene expression analysis, we found that either RA or RARalphaS77A induces many of the same differentiation response pathways and signaling molecules involved in U2OS cell differentiation. In addition, overexpression of the fibroblast growth factor 8f (FGF8f), one of the downstream targets induced by both RA and RARalphaS77A in U2OS cells, inhibits proliferation and induces expression of osteoblastic differentiation regulators. Hence, these data strongly suggest that RA-suppressed phosphorylation of RARalpha induces FGF8f expression to mediate differentiation response pathway in U2OS osteosarcoma cells.


Endothelial glycocalyx-the battleground for complications of sepsis and kidney injury.
After briefly discussing endothelial glycocalyx and its role in vascular physiology and renal disease, this overview focuses on its degradation very early in the course of microbial sepsis. We describe our recently proposed mechanism for glycocalyx degradation induced by exocytosis of lysosome-related organelles and release of their cargo. Notably, an intermediate in nitric oxide synthesis, NG-hydroxy-l-arginine, shows efficacy in curtailing exocytosis of these organelles and improvement in animal survival. These data not only depict a novel mechanism responsible for very early glycocalyx degradation, but may also outline a potential preventive therapy. The second issue discussed in this article is related to the therapeutic acceleration of restoration of already degraded endothelial glycocalyx. Here, using as an example our recent findings obtained with sulodexide, we illustrate the importance of the expedited repair of degraded endothelial glycocalyx for the survival of animals with severe sepsis. These two focal points of the review on glycocalyx may not only have broader disease applicability, but they may also provide additional evidence to buttress the idea of the importance of endothelial glycocalyx and its maintenance and repair in the prevention and treatment of an array of renal and nonrenal diseases.


Emergence and spread of carbapenem-resistant strains of Acinetobacter baumannii in a tertiary-care hospital in Poland.
This study analysed the occurrence of carbapenem resistance among Acinetobacter baumannii isolates from a tertiary-care hospital in Poland, together with the molecular epidemiology of these isolates and the risk-factors for their acquisition and possible nosocomial spread. The medical charts of 21 patients with Acinetobacter infection or colonisation revealed that A. baumannii isolates were obtained most frequently from intensive care unit and surgical patients (particularly those receiving transplantation surgery). First isolation occurred, on average, on day 21 following admission (range 5-45 days). Infection with Acinetobacter contributed directly to the death of seven patients. Several patients were infected with more than one strain, and molecular typing revealed the co-circulation of three predominant clones, of which two belonged to the Acinetobacter lineages designated as European clones I and II. All three clones encoded an OXA-51-type carbapenemase, but were negative for carbapenemases belonging to the OXA-23, OXA-24 and OXA-58 families. The OXA-51 gene was found in both resistant and susceptible isolates, and was not associated directly with carbapenem resistance. Etests with imipenem and imipenem plus EDTA indicated production of a metallo-beta-lactamase (MBL) in carbapenem-resistant isolates. PCRs for IMP-type MBLs were negative, but PCR using consensus primers for VIM-type MBLs were positive for carbapenem-resistant isolates belonging to the European clone II lineage. The occurrence of a VIM-type MBL in association with one of the epidemic lineages of A. baumannii is a cause for concern. Further studies are needed to evaluate possible inter-hospital spread of resistant A. baumannii strains in Poland.


Multiple treatment comparison in narcolepsy: a network meta-analysis.
Randomized controlled trials (RCTs) that compared the safety and efficacy of medical treatments for narcolepsy were analyzed using network meta-analysis. The RCTs in narcolepsy were searched. Network meta-analysis compared efficacy and safety of multiple treatments, multi-arm studies, and multi-criteria treatment decisions, based on a random model that assumed heterogeneity between studies, with corrections for multi-arm studies. Fourteen RCTs, three drug treatments, and six doses were identified: sodium oxybate (6 and 9 g/d), modafinil (between 200 and 400 mg/d), and pitolisant (up to 20 and up to 40 mg/d). Significant heterogeneity (>50%) between studies was found in 12/14 studies for almost all endpoints, but between-design consistency was present. For ESS and MWT, sodium oxybate 9 g/d, modafinil, and pitolisant up to 40 mg/d had similar efficacy. Pitolisant 40 mg/d and sodium oxybate 9 g/d in two nightly doses had similar efficacy in reducing cataplexy. A good safety profile characterized by a TEAE incidence risk ratio (IRR) <1.5 was found for all the compared treatments, except for sodium oxybate 9 g/d. Although no significant difference was found, Pitolisant 40 mg was shown with the best P scores for the benefit/risk (BR) ratio. Modafinil (200-400 mg/d), sodium oxybate 9 g/d, and pitolisant up to 40 mg/d had similar efficacy in reducing excessive day time sleepiness. Only sodium oxybate 9 g/d and pitolisant up to 40 mg/d were shown with a comparable beneficial effect on cataplexy. Overall, Pitolisant was found with the best P score on the BR ratio. PROSPERO 2017 CRD42017054686. Efficacy, safety, and benefit-risk comparison of alternative treatments in narcolepsy: a network multiple comparisons of treatment meta-analysis. http://www.crd.york.ac.uk/PROSPERO/display_record.php?ID=CRD42017054686.


Assessing change in chronic pain severity: the chronic pain grade compared with retrospective perceptions.
There is no standard method of measuring change in chronic pain severity. Clinical trials commonly use serial assessment scales, completed at two points in time, to estimate change in pain severity, while clinicians usually ask patients to make a retrospective assessment of change. How the two methods compare is not known. To assess different methods of measuring change in chronic pain severity, by comparing changes in scores on a serial measure of chronic pain severity using the Chronic Pain Grade (CPG) questionnaire and responders' retrospective perception of change in pain severity. Postal self-completion questionnaires. The Grampian region of Scotland. Postal questionnaires were sent in March and September 1998 to a random sample of 535 adults with chronic pain, drawn from responders to a postal survey of the region conducted in 1996. Corrected response rates of 87.5% and 90.7% were obtained. Over a six-month period poor levels of agreement were found, with responders' retrospective perceptions mirroring recorded changes in 41.8% of individuals (kappa = 0.081). A low partial correlation coefficient between the two measures (-0.209) was also found. Over a two-year period there were again poor levels of agreement, with responders' retrospective perceptions mirronng recorded changes in 35.2% of individuals (kappa = 0.071). A low partial correlation coefficient (-0.401) was again found. There was poor agreement and low correlation between two commonly used methods for assessing change in pain severity over time. This finding has important implications for both service practitioners and researchers.


The value of contrast-enhanced ultrasound in the characterization of vascular pattern of solid pancreatic lesions.
The AIM of our study was to evaluate the accuracy of CEUS in the characterization of pancreatic solid lesions, considering cross sectional imaging techniques (CE-CT/MRI) as the "gold standard" methods. We performed a retrospective, monocentric study that included 91 solid pancreatic lesions which were evaluated by CEUS and by a second-line contrast imaging technique (CT or MRI), considered as the reference method. The rate of a conclusive diagnosis based on a typical enhancement pattern was 94% (78/83 cases). In 72 cases out of 83 (86.7%) there was a perfect concordance between CEUS and the "gold-standard" imaging method (CE-CT/MRI). In our study, 88% (73/83) of the pancreatic lesions were categorized as malignant due to their typical wash-out aspect in the late phase. The overall accuracy of CEUS for the differential diagnosis of solid pancreatic tumors was approximately 81%. The accuracy of CEUS for the diagnosis of hypoenhancing pancreatic tumors was approximately 89.1%; while for the diagnosis of hyperenhancing pancreatic tumors it was approximately 72.8%. CEUS allows the differentiation between hypo- vs. hyperenhancing pancreatic solid lesions, with a considerable diagnostic accuracy, a fundamental step in the precise diagnosis of pancreatic tumors.


Protein binding of N-2-mercaptoethyl-1,3-diaminopropane via mixed disulfide formation after oral administration of WR 2721.
Earlier studies have shown that WR 2721 [H2N-(CH2)3-NH(CH2)2SPO3H2] is converted to its free thiol form, N-2-mercaptoethyl-1,3-diaminopropane (MDP), at the acidic pH of the stomach. MDP is a radioprotective compound and a mucolytic agent capable of decreasing sputum viscosity in the lungs of patients with cystic fibrosis. Conversion of WR 2721 and MDP to the corresponding sulfonic acid (MDP-SO3H) permits quantitative determination of these compounds in physiological fluids by use of an automatic amino acid analyzer. After oral administration of WR 2721 to human patients and rabbits it is converted to MDP and the free thiol form of the drug associates with plasma proteins by mixed disulfide linkage. The plasma proteins serve as a depot and reservoir of MDP for potential exchange at the tissues. When incubated with whole sputum or with purified mucin solutions in vitro, MDP decreased the viscosity of these solutions by reduction of the accessible disulfide bonds of the mucin molecule and was subsequently found in mixed disulfide association with the mucin molecule. The association of MDP with proteins via mixed disulfide linkage has important implications for the development of optimal dose regimens for administration of WR 2721 to patients.