Gene Introgression in Weeds Depends on Initial Gene Location in the Crop: Brassica napus-Raphanus raphanistrum Model.
The effect of gene location within a crop genome on its transfer to a weed genome remains an open question for gene flow assessment. To elucidate this question, we analyzed advanced generations of intergeneric hybrids, derived from an initial pollination of known oilseed rape varieties (Brassica napus, AACC, 2n = 38) by a local population of wild radish (Raphanus raphanistrum, RrRr, 2n = 18). After five generations of recurrent pollination, 307 G5 plants with a chromosome number similar to wild radish were genotyped using 105 B. napus specific markers well distributed along the chromosomes. They revealed that 49.8% of G5 plants carried at least one B. napus genomic region. According to the frequency of B. napus markers (0-28%), four classes were defined: Class 1 (near zero frequency), with 75 markers covering ∼70% of oilseed rape genome; Class 2 (low frequency), with 20 markers located on 11 genomic regions; Class 3 (high frequency), with eight markers on three genomic regions; and Class 4 (higher frequency), with two adjacent markers detected on A10. Therefore, some regions of the oilseed rape genome are more prone than others to be introgressed into wild radish. Inheritance and growth of plant progeny revealed that genomic regions of oilseed rape could be stably introduced into wild radish and variably impact the plant fitness (plant height and seed number). Our results pinpoint that novel technologies enabling the targeted insertion of transgenes should select genomic regions that are less likely to be introgressed into the weed genome, thereby reducing gene flow.


Bit-rate variable DPSK demodulation based on cascaded four-wave mixing.
We report a demodulator for DPSK signals at variable bit rates based on cascaded four-wave mixing (FWM). The demodulation utilizes two FWM processes in a photonic crystal fiber (PCF) with in-between dispersion in a chirped fiber Bragg grating (CFBG). The first FWM generates a wavelength-tunable idler carrying phase information of the signal. A tunable optical delay between the signal and the idler is then introduced by dispersion. The signal, the idler, and the pump are reflected by the CFBG with a reflectance of 99% back to the PCF to initiate the second FWM process. In the second FWM, the phase relationship between the signal and the one-bit-delayed idler determines an amplification or attenuation of the idler, converting phase modulation to intensity modulation. Error-free demodulations have been successfully demonstrated for both NRZ and RZ-DPSK signals at 5 and 10 Gb/s.


TI17, a novel compound, exerts anti-MM activity by impairing Trip13 function of DSBs repair and enhancing DNA damage.
Thyroid hormone receptor interacting protein 13 (Trip13) is an AAA-ATPase that regulates the assembly or disassembly protein complexes and mediates Double-strand breaks (DSBs) repair. Overexpression of Trip13 has been detected in many cancers and is associated with myeloma progression, disease relapse and poor prognosis inmultiple myeloma (MM). We have identified a small molecular, TI17, through a parallel compound-centric approach, which specifically targets Trip13. To identify whether TI17 targeted Trip13, pull-down and nuclear magnetic resonance spectroscopy (NMR) assays were performed. Cell counting kit-8, clone formation, apoptosis and cell cycle assays were applied to investigate the effects of TI17. We also utilized a mouse model to investigate the effects of TI17 in vivo. TI17 effectively inhibited the proliferation of MM cells, and induced the cycle arrest and apoptosis of MM cells. Furthermore, treatment with TI17 abrogates tumor growth and has no apparent side effects in mouse xenograft models. TI17 specifically impaired Trip13 function of DSBs repair and enhanced DNA damage responses in MM. Combining with melphalan or HDAC inhibitor panobinostat triggers synergistic anti-MM effect. Our study suggests that TI17 could be acted as a specific inhibitor of Trip13 and supports a preclinical proof of concept for therapeutic targeting of Trip13 in MM.


The AAA-ATPase Cdc48 and cofactor Shp1 promote chromosome bi-orientation by balancing Aurora B activity.
The assembly, disassembly and dynamic movement of macromolecules are integral to cell physiology. The ubiquitin-selective chaperone Cdc48 (p97 in Metazoa), an AAA-ATPase, might facilitate such processes in the cell cycle. Cdc48 in budding yeast was initially isolated from a mitotic mutant. However, its function in mitosis remained elusive. Here we show that the temperature-sensitive cdc48-3 mutant and depletion of cofactor Shp1 (p47 in Metazoa) cause cell-cycle arrest at metaphase. The arrest is due to a defect in bipolar attachment of the kinetochore that activates the spindle checkpoint. Furthermore, Cdc48-Shp1 positively regulates Glc7/protein phosphatase 1 by facilitating nuclear localization of Glc7, whereas it opposes Ipl1/Aurora B kinase activity. Thus, we propose that Cdc48-Shp1 promotes nuclear accumulation of Glc7 to counteract Ipl1 activity. Our results identify Cdc48 and Shp1 as critical components that balance the kinase and phosphatase activities at the kinetochore in order to achieve stable bipolar attachment.


Lymphotoxin-alpha gene 252A>G and metabolic syndrome features in Korean men with coronary artery disease.
Associations between variations in the lymphotoxin-alpha (LTA) gene and coronary artery disease (CAD) and type 2 diabetes have previously been reported. We investigated the influence of the LTA 252A>G and 804C>A polymorphisms on circulating parameters related to metabolic syndrome in Korean patients with CAD. The study subjects comprised 446 Korean male patients with CAD (age 53.9 y, BMI 25.2 kg/m2). The LTA 252A>G and 804C>A polymorphisms were almost in complete linkage disequilibrium (R(2)=99.4%). The LTA 252A>G polymorphism was associated with LDL particle size (P=0.046), HOMA-IR (P=0.022) and circulating levels of triglyceride (P=0.006), HDL-cholesterol (P=0.008), apo A1 (P=0.031), insulin (P=0.046), and adiponectin (P=0.018), after adjustment for BMI. CAD patients with LTA 252G/G (n=96) had a lower concentration of HDL-cholesterol, a smaller LDL particle size, and a higher triglyceride level, compared to those with 252A/A (n=121) or 252A/G (n=229). In addition, CAD patients with LTA 252G/G had lower concentrations of adiponectin and higher levels of insulin, and HOMA-IR than those with 252A/A. Homozygosity for rare alleles of the LTA 252A>G polymorphisms was associated with features of metabolic syndrome such as hyperinsulinemia, dyslipidemia, small LDL particle and low adiponectin level in CAD patients, suggesting that CAD patients with LTA 252GG are at high risk and needed an intensive intervention against further progression.


Two novel cytochrome P450 genes CYP6CS1 and CYP6CW1 from Nilaparvata lugens (Hemiptera: Delphacidae): cDNA cloning and induction by host resistant rice.
Two novel full-length P450 cDNAs, CYP6CS1 and CYP6CW1, were cloned from the fourth instar nymphs of brown planthopper Nilaparvata lugens Stål (Hemiptera: Delphacidae) reared on its susceptible rice variety Taichung Native 1 (TN1) plants. The deduced proteins are typical microsomal P450s sharing conserved structural and functional domains with other insect CYP6 members. Temporal expression analysis by northern blot hybridization indicated pre-exposure to N. lugens moderately resistant rice Minghui 63 (MH63) seedlings caused a time course-dependent induction of CYP6CS1 which peaked after 24 h of treatment; in contrast, CYP6CW1 was induced and remained at a constant time course from 0-72 h. CYP6CS1 and CYP6CW1 are dramatically induced in gut tissues and, slightly upregulated in carcass and fat bodies as revealed in spatial gene expression analysis. Whole mount in situ hybridizaion revealed that the two genes are expressed at a basal level in gut tissue and Malpighian tubules in nymphs fed with TN1 rice. After exposure to MH63, the expression of CYP6CW1 was found to be high in the whole gut, including Malpighian tubules. Expression of CYP6CS1 was significantly increased in midgut, and slightly increased in foregut, hindgut and Malpighian tubules. These data suggest a potential role of the two P450s in determining patterns of N. lugens-rice relationships through allelochemical detoxification.


A prospective intervention study to identify drug-related emergency department visits comparing a standard care group and a pharmaceutical care group.
Adverse drug reactions impose a major burden. Those adverse drug reactions might lead to hospitalization but are often not correctly identified in the emergency department (ED). Clinical pharmacists, although not routinely implemented, can help identify adverse drug reactions. The primary objective was to examine the drug association of ED visits in a pharmaceutical group with a clinical pharmacist integrated in the ED team compared with a standard group without additional support. This prospective intervention study was performed in the ED of a tertiary care university hospital in Leipzig, Germany. Patients who were ≥50 years old were included. From 1 March 2020 to May 31, 2020 patients were enrolled in the standard group. From 1 March 2021 to 31 May 2021, the pharmaceutical group was enrolled. The clinical pharmacist supported the ED team with patient´s detailed medication history and medication analysis. In both groups, patients were evaluated whether their ED visit was drug-related. The number of identified drug-related ED presentations were compared between the two groups. Interventions performed on adverse drug reaction management, causative drugs and patient characteristics were evaluated. A total of 798 patients were enrolled in the standard group and 827 patients in the pharmaceutical group. Patients whose ED visit was drug-related had a median age of 77 years [(Q25-Q75) 63.5-83.5] and took 7 [(Q25-Q75) 5-8] drugs in standard group. In the pharmaceutical group median age was 78 years [(Q25-Q75) 66-83] and number of drugs taken was 9 [(Q25-Q75) 5.25-11]. 31 (3.9%) drug-related ED visits were identified in the standard group compared to 104 (12.6%) in the pharmaceutical group (OR 3.56; 95% CI 2.35-5.38). An intervention on the patient's pharmacotherapy was performed in 16 drug-related ED visits in standard group compared to 77 in the pharmaceutical group. In this study the implementation of a clinical pharmacist was associated with improved identification of drug-related ED visits. Discontinuations of causal medications and dose reductions were significantly higher in the pharmaceutical group compared to the standard care group.


The clinicopathological significance of UBE2C in breast cancer: a study based on immunohistochemistry, microarray and RNA-sequencing data.
Ubiquitin-conjugating enzyme E2C (UBE2C) has been previously reported to correlate with the malignant progression of various human cancers, however, the exact molecular function of UBE2C in breast carcinoma (BRCA) remained elusive. We aimed to investigate UBE2C expression in BRCA and its clinical significance. The expression of UBE2C in 209 BRCA tissue samples and 53 adjacent normal tissue samples was detected using immunohistochemistry. The clinical role of UBE2C was analyzed. Public databases including the human protein atlas and Oncomine were used to assess UBE2C expression in BRCA. Moreover, the cancer genome atlas (TCGA) database was employed to investigate the prognostic value of UBE2C in BRCA. The positive expression rate of UBE2C in BRCA was 70.8% (148/209), and UBE2C expression in the adjacent breast tissue was negative. The expression of UBE2C was positively correlated with tumor size (r = 0.32, P < 0.001), histological grade (r = 0.237, P = 0.001), clinical stage (r = 0.198, P = 0.004), lymph node metastasis (r = 0.155, P = 0.026), HER2 expression level (r = 0.356, P < 0.001), Ki-67 expression level (r = 0.504, P < 0.001), and P53 expression level (r = 0.32, P = 0.001). Negative correlations were found between UBE2C expression and the ER (r = - 0.403, P < 0.001) and PR (r = - 0.468, P < 0.001) status. UBE2C gene expression data from the public databases all proved that UBE2C was overexpressed in BRCA. According to the TCGA data analysis, a higher positive expression of UBE2C was associated with worse survival of BRCA patients (P = 0.0428), and data from cBioPortal indicated that 11% of all sequenced BRCA patients possessed a gene alteration of UBE2C, predominately gene amplification and mRNA regulation. Ubiquitin-conjugating enzyme E2C might pose an oncogenic effect on the progression of BRCA.


Brain tumor segmentation using holistically nested neural networks in MRI images.
Gliomas are rapidly progressive, neurologically devastating, largely fatal brain tumors. Magnetic resonance imaging (MRI) is a widely used technique employed in the diagnosis and management of gliomas in clinical practice. MRI is also the standard imaging modality used to delineate the brain tumor target as part of treatment planning for the administration of radiation therapy. Despite more than 20 yr of research and development, computational brain tumor segmentation in MRI images remains a challenging task. We are presenting a novel method of automatic image segmentation based on holistically nested neural networks that could be employed for brain tumor segmentation of MRI images. Two preprocessing techniques were applied to MRI images. The N4ITK method was employed for correction of bias field distortion. A novel landmark-based intensity normalization method was developed so that tissue types have a similar intensity scale in images of different subjects for the same MRI protocol. The holistically nested neural networks (HNN), which extend from the convolutional neural networks (CNN) with a deep supervision through an additional weighted-fusion output layer, was trained to learn the multiscale and multilevel hierarchical appearance representation of the brain tumor in MRI images and was subsequently applied to produce a prediction map of the brain tumor on test images. Finally, the brain tumor was obtained through an optimum thresholding on the prediction map. The proposed method was evaluated on both the Multimodal Brain Tumor Image Segmentation (BRATS) Benchmark 2013 training datasets, and clinical data from our institute. A dice similarity coefficient (DSC) and sensitivity of 0.78 and 0.81 were achieved on 20 BRATS 2013 training datasets with high-grade gliomas (HGG), based on a two-fold cross-validation. The HNN model built on the BRATS 2013 training data was applied to ten clinical datasets with HGG from a locally developed database. DSC and sensitivity of 0.83 and 0.85 were achieved. A quantitative comparison indicated that the proposed method outperforms the popular fully convolutional network (FCN) method. In terms of efficiency, the proposed method took around 10 h for training with 50,000 iterations, and approximately 30 s for testing of a typical MRI image in the BRATS 2013 dataset with a size of 160 × 216 × 176, using a DELL PRECISION workstation T7400, with an NVIDIA Tesla K20c GPU. An effective brain tumor segmentation method for MRI images based on a HNN has been developed. The high level of accuracy and efficiency make this method practical in brain tumor segmentation. It may play a crucial role in both brain tumor diagnostic analysis and in the treatment planning of radiation therapy.


Progesterone induces dephosphorylation and inactivation of tyrosine hydroxylase in rat hypothalamic dopaminergic neurons.
After in vivo treatment, progesterone initially decreases tyrosine hydroxylase (TH) activity in the TIDA neurons, but subsequently increases TH activity with prolonged treatment. In order to explore the cellular mechanism for progesterone's effect, this study examined the acute inhibitory action of progesterone on TH activity in rat fetal hypothalamic dopaminergic neurons in vitro. Progesterone caused a rapid decrease in TH activity within 1 h, which was sustained for at least 6 h. However, the dopaminergic cells became refractory to progesterone with continuous treatment for 12 h to 10 days. Progesterone (10-100 nM) treatment suppressed TH activity in a concentration-dependent manner. The inhibitory effect of progesterone was dependent on prior exposure to estradiol. Whereas progesterone decreased TH activity, A ring-reduced metabolites of progesterone did not alter TH activity, suggesting that the response was specific to progesterone. Progesterone decreased radiolabeled phosphate incorporation into TH protein. Okadaic acid, a phosphoprotein phosphatase inhibitor, prevented the progesterone-induced suppression of TH activity and phosphate incorporation into TH, implicating dephosphorylation of TH as the cellular mechanism. In contrast, neither TH mRNA levels nor TH protein content was altered after 1 or 12 h of progesterone treatment. Progesterone decreased TH activity after pretreatment of the hypothalamic cells for 2 or 24 h with actinomycin D, an RNA synthesis inhibitor, suggesting that increased transcription does not mediate the effect. These data indicate that the acute progesterone-induced decline in TH activity is caused by dephosphorylation of TH.


The beta1 and beta3 integrins promote T cell receptor-mediated cytotoxic T lymphocyte activation.
Recognition by CD8+ cytotoxic T lymphocytes (CTLs) of antigenic peptides bound to major histocompatibility class (MHC) I molecules on target cells leads to sustained calcium mobilization and CTL degranulation resulting in perforin-dependent killing. We report that beta1 and beta3 integrin-mediated adhesion to extracellular matrix proteins on target cells and/or surfaces dramatically promotes CTL degranulation. CTLs, when adhered to fibronectin but not CTL in suspension, efficiently degranulate upon exposure to soluble MHC.peptide complexes, even monomeric ones. This adhesion induces recruitment and activation of the focal adhesion kinase Pyk2, the cytoskeleton linker paxillin, and the Src kinases Lck and Fyn in the contact site. The T cell receptor, by association with Pyk2, becomes part of this adhesion-induced activation cluster, which greatly increases its signaling.


C-terminal phosphorylation of Hsp70 and Hsp90 regulates alternate binding to co-chaperones CHIP and HOP to determine cellular protein folding/degradation balances.
Heat shock proteins Hsp90 and Hsp70 facilitate protein folding but can also direct proteins for ubiquitin-mediated degradation. The mechanisms regulating these opposite activities involve Hsp binding to co-chaperones including CHIP and HOP at their C-termini. We demonstrated that the extreme C-termini of Hsp70 and Hsp90 contain phosphorylation sites targeted by kinases including CK1, CK2 and GSK3-β in vitro. The phosphorylation of Hsp90 and Hsp70 prevents binding to CHIP and thus enhances binding to HOP. Highly proliferative cells contain phosphorylated chaperones in complex with HOP and phospho-mimetic and non-phosphorylable Hsp mutant proteins show that phosphorylation is directly associated with increased proliferation rate. We also demonstrate that primary human cancers contain high levels of phosphorylated chaperones and show increased levels of HOP protein and mRNA. These data identify C-terminal phosphorylation of Hsp70 and Hsp90 as a switch for regulating co-chaperone binding and indicate that cancer cells possess an elevated protein folding environment by the concerted action of co-chaperone expression and chaperone modifications. In addition to identifying the pathway responsible for regulating chaperone-mediated protein folding/degradation balances in normal cells, the data provide novel mechanisms to account for the aberrant chaperone activities observed in human cancer cells and have implications for the application of anti-chaperone therapies in cancer treatment.


Diagnostic and prognostic value of sCD14-ST--presepsin for patients admitted to hospital intensive care unit (ICU).
Sepsis is a serious problem in intensive care units all over the world. Biomarkers could be useful to identify patients at risk. We focused especially on the performance of presepsin (sCD14-ST), compared to C-reactive protein (CRP), procalcitonin (PCT) and CD64, to determine its diagnostic and prognostic indications. The study was conducted on 47 hospitalized patients after procedures, who were divided into three groups; systemic inflammatory response (SIRS), sepsis and septic shock. Expression of CD64 on neutrophils presented as CD64 index, sCD14-ST, CRP and PCT were measured in whole blood or plasma samples. All patients had standard samples like urine, respiratory tract samples etc. taken for culturing. Blood cultures were drawn to confirm bloodstream infection. Forty (85 %) patients had SIRS with bacterial infection and seven (15 %) patients had SIRS with no infection. All infections were confirmed with blood cultures. Biomarkers were evaluated in all patients. In patients with confirmed infection the values were high. The patients with bacterial infection showed statistical significance with CD64 index (p = 0.003), CRP (p = 0.049) and sCD14-ST (p = 0.026), but not with PCT (p = 1.000). The severity of diagnosed SIRS was significant only with PCT (p < 0.001). CD64 index, CRP and sCD14-ST served as good parameters to determine possible infection in patients that needed intensive care after major procedures. Values of PCT were the only ones to predict SIRS severity and could distinguish between sepsis and severe sepsis or septic shock.


Recent achievement in gene cloning and functional genomics in soybean.
Soybean is a model plant for photoperiodism as well as for symbiotic nitrogen fixation. However, a rather low efficiency in soybean transformation hampers functional analysis of genes isolated from soybean. In comparison, rapid development and progress in flowering time and photoperiodic response have been achieved in Arabidopsis and rice. As the soybean genomic information has been released since 2008, gene cloning and functional genomic studies have been revived as indicated by successfully characterizing genes involved in maturity and nematode resistance. Here, we review some major achievements in the cloning of some important genes and some specific features at genetic or genomic levels revealed by the analysis of functional genomics of soybean.


[PH3 residues in hazelnuts, soybeans and wheat following phosphine fumigation with non-constant concentrations].
In model tests hazelnuts, soy beans and wheat were fumigated with phosphine (PH3) at non constant concentrations. The influence of different concentration characteristics on the fumigation and the decomposition of phosphine residues was investigated in accordance with the fumigation technique. At the beginning the concentration increases, and after attaining the maximum gradually decreases to zero. The level of residues during the fumigation as well as the behaviour of residues during the storage of the fumigated products was monitored with a gas chromatographic method. The residues correlate with the concentration of phosphine, they also pass through a peak. The rate of decomposition of residues which had been formed in the phase of increasing concentration is greater than the rate of residues of equal magnitude which had been formed during the decreasing phase. When the concentration is even the maximum residue occurs later than the maximum concentration; when there is a steep trend both maximums coincide. This behaviour can be explained by the sorption and diffusion of phosphine. A comparison is made with the phosphine concentration which occurs during fumigation in practice. The parameters which produce a constant concentration trend with only one maximum and a non constant trend with an often increasing and decreasing concentration are discussed. The different behaviour of residues in these cases is described. Conclusions are drawn for the practice of fumigation.


DNA methyltransferase inhibitor suppresses fibrogenetic changes in human conjunctival fibroblasts.
This study aimed to clarify the effects of a DNA methyltransferase inhibitor on fibrogenetic changes in human conjunctival fibroblasts (HConF). HConF were pretreated with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-dC) for 48 h. After one passage, the cells were treated with 5 ng/ml of transforming growth factor (TGF)-β2 for 48 h, and the expression levels of α-smooth muscle actin (α-SMA), extracellular matrix proteins, and phosphorylated Smad3 were evaluated with western blotting. A fusion construct between the COL1A2 promoter and the luciferase gene was introduced into the HConF after the first passage, and the construct's activity was detected via a luciferase reporter gene assay. TGF-β2-induced upregulation of α-SMA was suppressed by pretreatment with 5-Aza-dC (0.1, 1.0, and 10 μM) in a dose-dependent manner. Upregulation of type I collagen was also suppressed by 10 μM 5-Aza-dC pretreatment. In contrast, 5-Aza-dC had no inhibitory effect on the expression of fibronectin or phosphorylated Smad3. However, COL1A2 promoter activity was suppressed with 5-Aza-dC pretreatment. In HConF, fibrogenetic changes were partly suppressed with a DNA methyltransferase inhibitor, suggesting an indirect inhibitory effect of the inhibitor on the COL1A2 promoter in HConF.


Preclinical and clinical advances in transposon-based gene therapy.
Transposons derived from Sleeping Beauty (SB), piggyBac (PB), or Tol2 typically require cotransfection of transposon DNA with a transposase either as an expression plasmid or mRNA. Consequently, this results in genomic integration of the potentially therapeutic gene into chromosomes of the desired target cells, and thus conferring stable expression. Non-viral transfection methods are typically preferred to deliver the transposon components into the target cells. However, these methods do not match the efficacy typically attained with viral vectors and are sometimes associated with cellular toxicity evoked by the DNA itself. In recent years, the overall transposition efficacy has gradually increased by codon optimization of the transposase, generation of hyperactive transposases, and/or introduction of specific mutations in the transposon terminal repeats. Their versatility enabled the stable genetic engineering in many different primary cell types, including stem/progenitor cells and differentiated cell types. This prompted numerous preclinical proof-of-concept studies in disease models that demonstrated the potential of DNA transposons for ex vivo and in vivo gene therapy. One of the merits of transposon systems relates to their ability to deliver relatively large therapeutic transgenes that cannot readily be accommodated in viral vectors such as full-length dystrophin cDNA. These emerging insights paved the way toward the first transposon-based phase I/II clinical trials to treat hematologic cancer and other diseases. Though encouraging results were obtained, controlled pivotal clinical trials are needed to corroborate the efficacy and safety of transposon-based therapies.


Delineation of the function of a major gamma delta T cell subset during infection.
Gammadelta T cells play important but poorly defined roles in pathogen-induced immune responses and in preventing chronic inflammation and pathology. A major obstacle to defining their function is establishing the degree of functional redundancy and heterogeneity among gammadelta T cells. Using mice deficient in Vgamma1+ T cells which are a major component of the gammadelta T cell response to microbial infection, a specific immunoregulatory role for Vgamma1+ T cells in macrophage and gammadelta T cell homeostasis during infection has been established. By contrast, Vgamma1+ T cells play no significant role in pathogen containment or eradication and cannot protect mice from immune-mediated pathology. Pathogen-elicited Vgamma1+ T cells also display different functional characteristics at different stages of the host response to infection that involves unique and different populations of Vgamma1+ T cells. These findings, therefore, identify distinct and nonoverlapping roles for gammadelta T cell subsets in infection and establish the complexity and adaptability of a single population of gammadelta T cells in the host response to infection that is not predetermined, but is, instead, shaped by environmental factors.


Objective, but Not Subjective, Sleepiness is Associated With Inflammation in Sleep Apnea.
Objective and subjective measures of excessive daytime sleepiness (EDS) are only weakly associated. No study, however, has examined whether these two measures of EDS differ in terms of underlying mechanisms and prognostic value. Pro-inflammatory cytokines, that is, interleukin-6 (IL-6) appear to promote sleepiness/fatigue, while the stress hormone cortisol promotes vigilance. We hypothesized that objective sleepiness is associated with increased levels of IL-6 and decreased levels of cortisol. We studied 58 obstructive sleep apnea (OSA) patients with clinical EDS and/or cardiovascular comorbidities who underwent 8-hour in-lab polysomnography for four consecutive nights. Objective and subjective daytime sleepiness were measured by Multiple Sleep Latency Test (MSLT), Epworth Sleepiness Scale (ESS), and Stanford Sleepiness Scale (SSS), respectively. Twenty-four-hour profiles of IL-6 and cortisol levels were assessed on the fourth day. The agreement between objective and subjective EDS in OSA patients was fair (kappa = 0.22). Objective EDS (lower MSLT) in OSA patients was associated with significantly elevated 24-hour (β = -0.34, p = .01), daytime (β = -0.30, p = .02) and nighttime (β = -0.38, p < .01) IL-6 levels, and significantly decreased daytime (β = 0.35, p = .01) cortisol levels. In contrast, subjective EDS (higher ESS/SSS) was not associated with either elevated IL-6 levels or decreased cortisol levels. Our findings suggest that OSA with objective EDS is the more severe phenotype of the disorder associated with low-grade inflammation, a link to cardiometabolic morbidity and mortality. Compared to subjective EDS, objective EDS is a stronger predictor of OSA severity and may be useful in the clinical management of the disorder.


Tissue-Based Thrombolysis for Wake-Up Stroke With Basilar Artery Occlusion: A Case Report.
Stroke from basilar artery occlusion is associated with a poor natural history with high rates of death and disability. Intravenous thrombolysis administered within 4.5 hours of last known well time improves the odds of a good neurological outcome after ischemic stroke, including in patients with basilar artery occlusion. Thrombectomy for basilar artery occlusion has had mixed outcomes. The WAKE-UP randomized clinical trial demonstrated that administration of intravenous thrombolysis can benefit select patients with wake-up strokes whose brain MRI shows restricted diffusion but no accompanying T2 FLAIR change. We report a case of a wake-up acute ischemic stroke presenting with acute vertigo followed by progressive brainstem dysfunction from a basilar artery occlusion. The patient was successfully treated with intravenous thrombolysis beyond 4.5 hours of last known well and symptom discovery time according to an MRI tissue-based approach resulting in partial recanalization of her basilar artery and recovery to near normal. This case suggests that hyperacute MRI can serve as a tissue clock to select patients with wake-up stroke for acute reperfusion therapy even if they do not meet standard trial inclusion criteria, including patients with basilar artery occlusion.


Plasma cytokine levels reveal deficiencies in IL-8 and gamma interferon in Long-COVID.
Up to half of individuals who contract SARS-CoV-2 develop symptoms of long-COVID approximately three months after initial infection. These symptoms are highly variable, and the mechanisms inducing them are yet to be understood. We compared plasma cytokine levels from individuals with long-COVID to healthy individuals and found that those with long-COVID had 100% reductions in circulating levels of interferon gamma (IFNγ) and interleukin-8 (IL-8). Additionally, we found significant reductions in levels of IL-6, IL-2, IL-17, IL-13, and IL-4 in individuals with long-COVID. We propose immune exhaustion as the driver of long-COVID, with the complete absence of IFNγ and IL-8 preventing the lungs and other organs from healing after acute infection, and reducing the ability to fight off subsequent infections, both contributing to the myriad of symptoms suffered by those with long-COVID.


Seizure activity per se does not induce tissue damage markers in human neocortical focal epilepsy.
The contribution of recurring seizures to the progression of epileptogenesis is debated. Seizure-induced brain damage is not conclusively demonstrated either in humans or in animal models of epilepsy. We evaluated the expression of brain injury biomarkers on postsurgical brain tissue obtained from 20 patients with frequent seizures and a long history of drug-resistant focal epilepsy. The expression patterns of specific glial, neuronal, and inflammatory molecules were evaluated by immunohistochemistry in the core of type II focal cortical dysplasias (FCD-II), at the FCD boundary (perilesion), and in the adjacent normal-appearing area included in the epileptogenic region. We also analyzed surgical specimens from cryptogenic patients not presenting structural alterations at imaging. Astroglial and microglial activation, reduced neuronal density, perivascular CD3-positive T-lymphocyte clustering, and fibrinogen extravasation were demonstrated in the core of FCD-II lesions. No pathological immunoreactivity was observed outside the FCD-II or in cryptogenetic specimens, where the occurrence of interictal and ictal epileptiform activity was confirmed by either stereo-electroencephalography or intraoperative electrocorticography. Recurrent seizures do not induce the expression of brain damage markers in nonlesional epileptogenic cortex studied in postsurgical tissue from cryptogenic and FCD patients. This evidence argues against the hypothesis that epileptiform activity per se contributes to focal brain injury, at least in the neocortical epilepsies considered here. Ann Neurol 2017;82:331-341.


An in vivo molecular imaging probe (18)F-Annexin B1 for apoptosis detection by PET/CT: preparation and preliminary evaluation.
There is an increasing need to develop non-invasive molecular imaging strategies for visualizing and quantifying apoptosis status of diseases (especially for cancer) for diagnosis and monitoring treatment response. Since externalization of phosphatidylserine (PS) is one of the early molecular events during apoptosis, Annexin B1 (AnxB1), a member of Annexins family with high affinity toward the head group of PS, could be a potential positron emission tomography (PET) imaging probe for imaging cell death process after labeled by positron-emitting nuclides, such as (18)F. In the present study, we investigated a novel PET probe, (18)F-labeled Annexin B1 ((18)F-AnxB1), for apoptosis imaging. (18)F-AnxB1 was prepared reliably by conjugating AnxB1 with a (18)F-tag, N-succinimidyl 4-[(18)F]fluorobenzoate ([(18)F]SFB), in a radiolabeling yield of about 20 % within 40 min. The in vitro binding of (18)F-AnxB1 with apoptotic cells induced by anti-Fas antibody showed twofold increase compared to those without treatment, confirmed by flow cytometric analysis with AnxV-FITC/PI staining. Stability tests demonstrated (18)F-AnxB1 was rather stable in vitro and in vivo without degradation. The serial (18)F-AnxB1 PET/CT scans in healthy rats outlined its biodistribution and pharmacokinetics, indicating a rapid renal clearance and predominant accumulation into kidney and bladder at 2 h p.i. (18)F-AnxB1 PET/CT imaging was successfully applied to visualize in vivo apoptosis sites in tumor induced by chemotherapy and in kidney simulated by ischemia-reperfusion injury. The high-contrast images were obtained at 2 h p.i. to delineate apoptotic tumor. Apoptotic region could be still identified by (18)F-AnxB1 PET 4 h p.i., despite the high probe retention in kidneys. In summary, we have developed (18)F-AnxB1 as a PS-specific PET probe for the apoptosis detection and quantification which could have broad applications from disease diagnosis to treatment monitoring, especially in the cases of cancer.


Role of apoptosis in ventricular remodeling.
Cell loss is a common feature in the failing heart, and this contributes to the relentless progression seen in patients with heart failure. Apoptosis is one of the most common causes of cell loss in animals and humans with heart failure. There is increasing evidence that apoptosis, even while occurring in a low-grade manner, can mediate heart failure. Moreover, inhibiting apoptosis successfully prevents or attenuates heart failure in several animal models. More importantly, apoptosis is one of the few mechanisms that can be easily modulated using pharmacologic or gene therapy approaches. Animal data, obtained in the past few years, have proven the feasibility and success of this approach toward altering the natural history of heart failure. Human studies are pending, but a number of issues such as the type of inhibitor and its optimum timing/dose will need to be resolved before this becomes a reality. Nevertheless, these data will accrue over time, and anti-apoptotic therapy is likely to emerge as an important form of heart failure therapy.


Antimycobacterial activity of cerulenin and its effects on lipid biosynthesis.
Cerulenin is a potent inhibitor of fatty acid synthase (FAS) in a variety of prokaryotic and eukaryotic cells. Using a standardized mycobacterial susceptibility test, we have observed that cerulenin inhibits the growth of several species of mycobacteria, including tuberculous species such as Mycobacterium tuberculosis (H37Rv and clinical isolates) and Mycobacterium bovis BCG (hereafter called BCG), as well as several non-tuberculous species: Mycobacterium smegmatis, the Mycobacterium avium-intracellulare complex (MAC), Mycobacterium kansasii and others. All species and strains tested, including multi-drug resistant isolates of M. tuberculosis, were susceptible to cerulenin with MICs ranging from 1.5 to 12.5 mg/L. Two-dimensional thin-layer chromatography revealed different inhibition patterns of lipid synthesis between tuberculous and non-tuberculous mycobacteria. Cerulenin treatment resulted in a relative increase in phospholipids and mycolic acids in MAC and M. smegmatis, whereas in cerulenin-treated BCG, phospholipids and mycolic acids diminished relative to controls. In addition, long-chain extractable lipids (intermediate in polarity), triglycerides and glycopeptidolipids decreased with cerulenin treatment in all three species of mycobacteria tested. Qualitative changes in several of these lipid classes indicate inhibition in the synthesis of intermediate and long-chain fatty acids. Our results suggest that cerulenin's primary effect may be inhibition of intermediate and long-chain lipid synthesis, with little effect on the synthesis of other lipid classes. In addition, the BCG-specific reduction in phospholipids and mycolic acids suggests the presence of a unique cerulenin-sensitive FAS system in tuberculous mycobacteria. Since pathogenic mycobacteria produce novel long-chain fatty acids, inhibition of fatty acid synthesis offers a potential target for the development of antimycobacterial drugs.


Upper brachial plexus injuries: grafts vs ulnar fascicle transfer to restore biceps muscle function.
Nerve transfers or graft repairs in upper brachial plexus palsies are 2 available options for elbow flexion recovery. To assess outcomes of biceps muscle strength when treated either by grafts or nerve transfer. A standard supraclavicular approach was performed in all patients. When roots were available, grafts were used directed to proximal targets. Otherwise, a distal ulnar nerve fascicle was transferred to the biceps branch. Elbow flexion strength was measured with a dynamometer, and an index comparing the healthy arm and the operated-on side was developed. Statistical analysis to compare both techniques was performed. Thirty-five patients (34 men) were included in this series. Mean age was 28.7 years (standard deviation, 8.7). Twenty-two patients (62.8%) presented with a C5-C6 injury, whereas 13 patients (37.2%) had a C5-C6-C7 lesion. Seventeen patients received reconstruction with grafts, and 18 patients were treated with a nerve transfer from the ulnar nerve to the biceps. The trauma to surgery interval (mean, 7.6 months in both groups), strength in the healthy arm, and follow-up duration were not statistically different. On the British Medical Research Council muscle strength scale, 8 of 17 (47%) patients with a graft achieved ≥ M3 biceps flexion postoperatively, vs 16 of 18 (88%) post nerve transfers (P = .024). This difference persisted when a muscle strength index assessing improvement relative to the healthy limb was used (P = .031). The results obtained from ulnar nerve fascicle transfer to the biceps branch were superior to those achieved through reconstruction with grafts.


Alteration of extracellular matrix modulators after nonablative laser therapy in skin rejuvenation.
Nonablative laser therapy is widely practised for skin rejuvenation, which stimulates collagen production and dermal matrix remodelling. Matrix remodelling is primarily modulated by a coordinated action of matrix metalloproteinases (MMPs) and their inhibitors, but the effects of nonablative lasers on these matrix modulators are not fully investigated. To evaluate the changes in matrix modulators, such as MMP-1, MMP-2, MMP-3, MMP-9 and MT1-MMP, and their inhibitors (TIMP-1, TIMP-2 and RECK in particular), after nonablative laser treatments of human facial skin. Twenty-four adult volunteers received a series of four nonablative laser treatments separated by 3-week intervals on facial skin. Two-millimetre skin punch biopsies were obtained at baseline and 3 weeks after the last treatment. Nonablative laser treatments led to a robust increase in two major dermal matrix components, type I collagen and tropoelastin. Among MMPs tested, levels of MMP-2 mRNA were statistically significantly increased, but the amount of active MMP-2 was rather reduced. More importantly, the expression level of RECK was significantly enhanced by laser treatments. Clinical outcomes following nonablative laser treatments may result not only from increased biosynthesis but also from decreased degradation, via an induction of RECK expression, of matrix proteins.


Inhaled Medication Use in Smokers With Normal Spirometry.
The objective of our study was to identify variables associated with inhaled medication use in smokers with normal spirometry (GOLD-0) and to examine the association of inhaled medication use with development of exacerbations and obstructive spirometry in the future. We performed a retrospective multivariable analysis of GOLD-0 subjects identified in data from the COPDGene study to examine factors associated with medication use. Five categories were identified: (1) no medications, (2) short-acting bronchodilator, (3) long-acting bronchodilator; long-acting muscarinic antagonists and/or long-acting β agonist, (4) inhaled corticosteroids (ICS) with or without long-acting bronchodilator, and (5) dual bronchodilator with ICS. Sensitivity analysis was performed excluding subjects with history of asthma. We also evaluated whether long-acting inhaled medication use was associated with exacerbations and obstructive spirometry at the follow-up visit 5 y after enrollment. Of 4,303 GOLD-0 subjects within the analysis, 541 of them (12.6%) received inhaled medications. Of these, 259 (6%) were using long-acting inhaled medications and 282 (6.6%) were taking short-acting bronchodilator. Female sex (odds ratio [OR] 1.47, P = .003), numerous medical comorbidities, radiographic emphysema (OR 2.22, P = .02), chronic bronchitis (OR 1.77, P < .001), dyspnea (OR 2.24, P < .001), asthma history (OR 15.56, P < .001), prior exacerbation (OR 8.45, P < .001), and 6-min walk distance (OR 0.9, P < .001) were associated with medication use. Minimal changes were noted in a sensitivity analysis. Additionally, inhaled medications were associated with increased total (incidence rate ratio 2.83, P < .001) and severe respiratory exacerbations (incidence rate ratio 3.64, P < .001) and presence of obstructive spirometry (OR 2.83, P = .002) at follow-up. Respiratory symptoms, history of asthma, and radiographic emphysema were associated with inhaled medication use in smokers with normal spirometry. These individuals were more likely to develop obstructive spirometry, which suggests that health care providers may be able to identify obstructive lung disease prior to meeting the current criteria for COPD.


A single point mutation controls the cholesterol dependence of Semliki Forest virus entry and exit.
Membrane fusion and budding are key steps in the life cycle of all enveloped viruses. Semliki Forest virus (SFV) is an enveloped alphavirus that requires cellular membrane cholesterol for both membrane fusion and efficient exit of progeny virus from infected cells. We selected an SFV mutant, srf-3, that was strikingly independent of cholesterol for growth. This phenotype was conferred by a single amino acid change in the E1 spike protein subunit, proline 226 to serine, that increased the cholesterol independence of both srf-3 fusion and exit. The srf-3 mutant emphasizes the relationship between the role of cholesterol in membrane fusion and virus exit, and most significantly, identifies a novel spike protein region involved in the virus cholesterol requirement.


[Medulloadrenal secretion and peripheral sympathomimetic receptors in familial dysautonomia].
A case of familial dysautonomia in a 30-year old woman with severe postural hypotension is reported. A study of adrenal medulla secretion showed that it was inadapted to the fall in blood pressure, with no increase in dopamine secretion. Insulin-induced hypoglycaemia had no effect on adrenal medulla secretion. Changes in blood pressure and heart rate were studied after administration of alpha- and beta-adrenergic drugs combined with fludrocortisone. The increase observed in blood pressure and heart rate suggests that peripheral catecholamine receptors are present in dysautonomic patients. The best treatment of postural hypotension in these patients seems to be a dihydroergotamine-fludrocortisone combination.


The short-term effectiveness of scapular focused taping on scapular movement in tennis players with shoulder pain: A within-subject comparison.
This study aimed to investigate the short-term effectiveness of scapular focused taping (SFT) on scapular position and kinematics during the tennis serve among professional players with and without shoulder pain. The cohort included 7 players who had no history of non-shoulder pain (NSP) and 6 players with shoulder pain (SP). All participants performed tennis flat serves while the Qualisys motion capture system recorded three-dimensional scapular kinematic data according to the International Society of Biomechanics recommendations. SFT was applied to the participants' torso aligned with the lower trapezius, and the same movements were repeated. In the SP group, the scapula was tilted more posteriorly after the application of SFT as compared to before at ball release and maximally externally rotated humerothoracic joint during tennis serve (t = -5.081, P = .004 and t = -2.623, P = .047, respectively). In the NSP group, the scapula was tilted more posteriorly with SFT as compared to without at first 75% timing of the cocking phase and maximally externally rotated humerothoracic joint (t = -3.733, P = .010 and t = -2.510, P = .046, respectively). And the SP group exhibited a more rotated scapula externally after the application of SFT as compared to before at Ball impact (t = 5.283, P = .003). SFT had a positive immediate effect on the scapular posterior tilting and external rotation during certain phases of the tennis serve among tennis athletes with and without shoulder pain. These findings may help clinicians and sports practitioners to prevent and rehabilitate shoulder injuries for overhead athletes. Level of evidence: Level III; Case-Control Design; Comparative Study.


Resonant out-of-phase fluorescence microscopy and remote imaging overcome spectral limitations.
We present speed out-of-phase imaging after optical modulation (OPIOM), which exploits reversible photoswitchable fluorophores as fluorescent labels and combines optimized periodic illumination with phase-sensitive detection to specifically retrieve the label signal. Speed OPIOM can extract the fluorescence emission from a targeted label in the presence of spectrally interfering fluorophores and autofluorescence. Up to four fluorescent proteins exhibiting a similar green fluorescence have been distinguished in cells either sequentially or in parallel. Speed OPIOM is compatible with imaging biological processes in real time in live cells. Finally speed OPIOM is not limited to microscopy but is relevant for remote imaging as well, in particular, under ambient light. Thus, speed OPIOM has proved to enable fast and quantitative live microscopic and remote-multiplexed fluorescence imaging of biological samples while filtering out noise, interfering fluorophores, as well as ambient light.Generally, fluorescence imaging needs to be done in a dark environment using molecules with spectrally separated emissions. Here, Quérard et al. develop a protocol for high-speed imaging and remote sensing of spectrally overlapping reversible photoswitchable fluorophores in ambient light.


Tr1 Cells Emerge and Suppress Effector Th17 Cells in Glomerulonephritis.
T regulatory type 1 (Tr1) cells, which are defined by their regulatory function, lack of Foxp3, and high expression of IL-10, CD49b, and LAG-3, are known to be able to suppress Th1 and Th17 in the intestine. Th1 and Th17 cells are also the main drivers of crescentic glomerulonephritis (GN), the most severe form of renal autoimmune disease. However, whether Tr1 cells emerge in renal inflammation and, moreover, whether they exhibit regulatory function during GN have not been thoroughly investigated yet. To address these questions, we used a mouse model of experimental crescentic GN and double Foxp3mRFP IL-10eGFP reporter mice. We found that Foxp3neg IL-10-producing CD4+ T cells infiltrate the kidneys during GN progression. Using single-cell RNA sequencing, we could show that these cells express the core transcriptional factors characteristic of Tr1 cells. In line with this, Tr1 cells showed a strong suppressive activity ex vivo and were protective in experimental crescentic GN in vivo. Finally, we could also identify Tr1 cells in the kidneys of patients with antineutrophil cytoplasmic autoantibody-associated GN and define their transcriptional profile. Tr1 cells are currently used in several immune-mediated inflammatory diseases, such as T-cell therapy. Thus, our study provides proof of concept for Tr1 cell-based therapies in experimental GN.


Contingency Management and Deliberative Decision-Making Processes.
Contingency management is an effective treatment for drug addiction. The current explanation for its success is rooted in alternative reinforcement theory. We suggest that alternative reinforcement theory is inadequate to explain the success of contingency management and produce a model based on demand curves that show how little the monetary rewards offered in this treatment would affect drug use. Instead, we offer an explanation of its success based on the concept that it accesses deliberative decision-making processes. We suggest that contingency management is effective because it offers a concrete and immediate alternative to using drugs, which engages deliberative processes, improves the ability of those deliberative processes to attend to non-drug options, and offsets more automatic action-selection systems. This theory makes explicit predictions that can be tested, suggests which users will be most helped by contingency management, and suggests improvements in its implementation.


Reported cystoscopic experience correlates poorly with objective assessment of cystoscopic skills.
Although gynecologists perform a large number of surgeries in close proximity to the ureters and the urinary bladder, traditionally, Obstetrics and Gynecology resident physicians are not formally taught to perform cystoscopy. The primary objective was to document resident physicians' performance in diagnostic cystoscopic instrumentation and technique. The secondary objective was to examine if reported prior cystoscopic experience was associated with superior performance. Fifty-one postgraduate year 4 residents with reported experience with cystoscopy were evaluated using an operation-specific checklist and a global ratings scale based on the Objective Structured Assessment of Technical Skill model. Before evaluation, they attended a formal training session in cystoscopy, which included practice on a bench model of a simulated bladder. Forty-three of the 51 residents were able to successfully perform a thorough diagnostic examination immediately after the course. Six of the 8 failures were re-evaluated 2 weeks later and successfully performed a complete examination at that time. Before the course, the residents had performed a mean of 12.2 cystoscopic examinations as the primary surgeon (median, 12; range, 2-33). The number of reported cystoscopic examinations performed before the course did not correlate with the ability to perform a thorough cystoscopic examination (r = -0.109; P = 0.496). For this group of residents, there was poor correlation between the number of reported cystoscopic examinations and the ability to perform diagnostic cystoscopy. Trainees may not be able to determine when they have received enough instruction in hands-on training with models before acquisition of technical skills.


Image sharpness assessment based on local phase coherence.
Sharpness is an important determinant in visual assessment of image quality. The human visual system is able to effortlessly detect blur and evaluate sharpness of visual images, but the underlying mechanism is not fully understood. Existing blur/sharpness evaluation algorithms are mostly based on edge width, local gradient, or energy reduction of global/local high frequency content. Here we understand the subject from a different perspective, where sharpness is identified as strong local phase coherence (LPC) near distinctive image features evaluated in the complex wavelet transform domain. Previous LPC computation is restricted to be applied to complex coefficients spread in three consecutive dyadic scales in the scale-space. Here we propose a flexible framework that allows for LPC computation in arbitrary fractional scales. We then develop a new sharpness assessment algorithm without referencing the original image. We use four subject-rated publicly available image databases to test the proposed algorithm, which demonstrates competitive performance when compared with state-of-the-art algorithms.


VHL loss in renal cell carcinoma leads to up-regulation of CUB domain-containing protein 1 to stimulate PKC{delta}-driven migration.
A common genetic mutation found in clear cell renal cell carcinoma (CC-RCC) is the loss of the von Hippel-Lindau (VHL) gene, which results in stabilization of hypoxia-inducible factors (HIFs), and contributes to cancer progression and metastasis. CUB-domain-containing protein 1 (CDCP1) was shown to promote metastasis in scirrhous and lung adenocarcinomas as well as in prostate cancer. In this study, we established a molecular mechanism linking VHL loss to induction of the CDCP1 gene through the HIF-1/2 pathway in renal cancer. Also, we report that Fyn, which forms a complex with CDCP1 and mediates its signaling to PKCδ, is a HIF-1 target gene. Mechanistically, we found that CDCP1 specifically regulates phosphorylation of PKCδ, but not of focal adhesion kinase or Crk-associated substrate. Signal transduction from CDCP1 to PKCδ leads to its activation, increasing migration of CC-RCC. Furthermore, patient survival can be stratified by CDCP1 expression at the cell surface of the tumor. Taken together, our data indicates that CDCP1 protein might serve as a therapeutic target for CC-RCC.


Perphenazine-Macrocycle Conjugates Rapidly Sequester the Aβ42 Monomer and Prevent Formation of Toxic Oligomers and Amyloid.
Alzheimer's disease is imposing a growing social and economic burden worldwide, and effective therapies are urgently required. One possible approach to modulation of the disease outcome is to use small molecules to limit the conversion of monomeric amyloid (Aβ42) to cytotoxic amyloid oligomers and fibrils. We have synthesized modulators of amyloid assembly that are unlike others studied to date: these compounds act primarily by sequestering the Aβ42 monomer. We provide kinetic and nuclear magnetic resonance data showing that these perphenazine conjugates divert the Aβ42 monomer into amorphous aggregates that are not cytotoxic. Rapid monomer sequestration by the compounds reduces fibril assembly, even in the presence of pre-formed fibrillar seeds. The compounds are therefore also able to disrupt monomer-dependent secondary nucleation, the autocatalytic process that generates the majority of toxic oligomers. The inhibitors have a modular design that is easily varied, aiding future exploration and use of these tools to probe the impact of distinct Aβ42 species populated during amyloid assembly.


Predictors of statin adherence, switching, and discontinuation in the USAGE survey: understanding the use of statins in America and gaps in patient education.
Although statins have been shown to reduce cardiovascular disease mortality, less than half of U.S. adults achieve their low-density lipoprotein cholesterol goal. In many patients initiated on a statin, adherence rates decrease over time. To characterize current and former statin users, identify reasons for the discontinuation or switching of statins, and identify factors associated with adherence. The USAGE survey is a cross-sectional, self-administered Internet-based survey of 10,138 U.S. adults fielded September to October 2011. The following statin users were identified and compared: adherent nonswitchers, adherent switchers, non-adherent switchers, and discontinuers. Univariate and multivariate models using a priori covariates for adherence and discontinuation were examined. Most participants were current statin users who adhered with their prescribed statin (82.5%, n = 8371). Former statin users or discontinuers (12%, n = 1220) cited muscle pain, a side effect, as the primary reason for discontinuation (60%), followed by cost (16%), and then perceived lack of efficacy (13%). Discontinuers were less satisfied with their physicians' explanation of cholesterol treatment, more likely to use the Internet to research statins, and less likely to undergo frequent cholesterol monitoring. Among adherent statin users, the primary reasons for switching were muscle side effects (33%) and cost (32%). Individuals at risk for non-adherence included those with low household income, those who experienced muscle pain as a side effect while on statin therapy, and those taking medication for cardiovascular disease. Statin-related muscle side effects are common and contribute significantly to rates of discontinuation, switching, and non-adherence. Improved physician patient communication about side effects and benefits of statins are necessary to improve both adherence and outcomes.


Access to divergent benzo-heterocycles via a catalyst-dependent strategy in the controllable cyclization of o-alkynyl-N-methoxyl-benzamides.
A chemo- and regio-selectively controllable approach for construction of diverse benzo-heterocycles is established. A new strategy for using the ligand effect in gold catalysis to control the regioselectivity in the cyclization of o-alkynyl-N-methoxyl-benzamide is successfully achieved. Meanwhile, the chemoselectivity between nitrogen and oxygen nucleophiles is precisely switched by gold and platinum catalysts.


Recording pattern visual evoked potentials under chloral hydrate sedation.
Accurate assessment of visual function in infants and preverbal children is difficult because of the dependence on vision tests that require the patient's cooperation. In an attempt to resolve this problem, we evaluated pattern visual evoked potentials (PVEPs) obtained on infants under chloral hydrate sedation. Ten patients were studied--four with unilateral amblyopia, three with suspected amblyopia, and three without amblyopia. Patients with amblyopia showed robust and reproducible responses from the sound eye, while responses from the amblyopic eye were significantly diminished (amplitude ratio less than or equal to 0.63). The PVEP responses from patients without amblyopia were large in amplitude and symmetrical between fellow eyes (amplitude ratio greater than 0.9). In two eyes with normal vision, optically fogging the patterned stimulus image substantially reduced the PVEP response, establishing a sensitivity to pattern images rather than changes in the overall luminance. We have thus determined that the PVEP can be reliably obtained under chloral hydrate sedation and that these responses, like PVEP responses taken in the awake state, reflect the presence of amblyopia and image clarity. Use of the PVEP with chloral hydrate sedation may prove clinically useful for detecting amblyopia in uncooperative infants and children.


Electrochemically Controlled RAFT Polymerization for Highly Sensitive Electrochemical Biosensing of Protein Kinase Activity.
Phosphorylation of proteins catalyzed by protein kinases (PKs) is essential to many biological processes; the sensitive detection of PK activity and the screening of PK inhibitors are thus integral to disease diagnosis and drug discovery. Herein, a highly sensitive biosensor has been fabricated for the electrochemical detection of PK activity by exploiting the electrochemically controlled reversible addition-fragmentation chain transfer (eRAFT) polymerization as a novel amplification strategy. The fabrication of the eRAFT-polymerization-based electrochemical biosensor involves (1) the immobilization of substrate peptides onto a gold electrode by way of gold-sulfur self-assembly, (2) the site-specific phosphorylation of substrate peptides by PKs, (3) the anchoring of carboxyl-group-containing chain transfer agents (CTAs) to the phosphorylated sites, and (4) the eRAFT polymerization under a potentiostatic condition, using ferrocenylmethyl methacrylate (FcMMA) as the monomer. Through the eRAFT polymerization, long polymer chains containing numerous electroactive Fc tags can be de novo grafted from each phosphorylated site, resulting in significant amplification of the electrochemical detection signal. The as-fabricated biosensor is highly selective and features a very low detection limit of 1.02 mU mL-1, in the presence of adenosine 3',5'-cyclic monophosphate (cAMP)-dependent PK (PKA) as the model target. Results also demonstrate that it can be applied to the screening of PK inhibitors and the detection of PK activity in complex serum samples and cell lysates. Moreover, it holds the merits of easy fabrication, high efficiency, and low cost, which make it a promising tool for the detection of PK activity and the screening of potential PK inhibitors.


Cardiovascular consequences of early-onset growth hormone excess.
Acromegaly has relevant effects on the cardiovascular system, but few data deal with the early effects of GH and IGF-I excess. To study the early stage of acromegalic cardiomyopathy and give indirect evidence of the mechanisms underlying GH and IGF-I action on the human heart, 25 patients with uncomplicated acromegaly [15 young subjects with short-term (< or =5 yr) disease and 10 with long-term (>5 yr) disease] and 25 sex- and age-matched controls were studied. Cardiovascular risk parameters were studied by standard methods; cardiac morphology by M-mode and Doppler echocardiography, cardiac function at rest and at peak exercise by equilibrium radionuclide angiography, and vascular disease at common carotid arteries by Doppler ultrasonography. In the patient group these measurements were repeated after 6 months of treatment with octreotide-LAR (20-40 mg, im, every 28 d). Glucose, glycosylated hemoglobin, insulin, low density lipoprotein cholesterol, triglycerides, and fibrinogen levels were higher, and high density lipoprotein cholesterol levels were lower in acromegalic patients than in controls. Resting blood pressure was similar in patients and controls, whereas heart rate at rest and systolic blood pressure at peak exercise were higher in the patients. The left ventricular mass index was higher in acromegalic patients than in controls (123.3 +/- 8.9 vs. 81.5 +/- 4.3 g/m(2); P < 0.001); seven patients had left ventricular hypertrophy. Diastolic function was similar in the two groups. The ejection fraction at rest, but not at peak exercise, was significantly increased in the patients compared with controls. As a consequence the exercise-induced changes in the ejection fraction were lower in patients than controls (8.7 +/- 1.1% vs. 21.9 +/- 3.5%; P < 0.001). At common carotid ultrasonography, young patients with acromegaly had increased diastolic peak velocity and increased intima media thickness, even if neither patient nor controls had atherosclerotic plaques. Six months after OCT-LAR treatment, GH and IGF-I levels remarkably decreased in all patients; 8 (53.3%) achieved disease control. Insulin, total cholesterol, and fibrinogen levels reduced, whereas high density lipoprotein cholesterol levels increased. Both at rest and at peak exercise, heart rate significantly decreased, whereas systolic and diastolic blood pressures did not change. The left ventricular mass index was significantly reduced, but it was still higher than the control value (101.6 +/- 3.5 g/m(2); P < 0.01). The left ventricular ejection fraction at rest was significantly reduced, but its response at peak exercise was increased (16.3 +/- 2.4%), becoming similar to the control value. At common carotids, the intima media thickness of right and left arteries was significantly reduced as was the diastolic peak velocity without any change in systolic peak velocity. Short-term GH excess, despite causing enhanced cardiac performance at rest, reduces cardiac performance on effort and impairs vascular morphology. These deleterious effects of early-onset acromegaly are ameliorated by suppressing GH/IGF-I levels for 6 months.


Acoustic markers of prominence influence infants' and adults' segmentation of speech sequences.
Two experiments investigated the way acoustic markers of prominence influence the grouping of speech sequences by adults and 7-month-old infants. In the first experiment, adults were familiarized with and asked to memorize sequences of adjacent syllables that alternated in either pitch or duration. During the test phase, participants heard pairs of syllables with constant pitch and duration and were asked whether the syllables had appeared adjacently during familiarization. Adults were better at remembering pairs of syllables that during familiarization had short syllables preceding long syllables, or high-pitched syllables preceding low-pitched syllables. In the second experiment, infants were familiarized and tested with similar stimuli as in the first experiment, and their preference for pairs of syllables was accessed using the head-turn preference paradigm.When familiarized with syllables alternating in pitch, infants showed a preference to listen to pairs of syllables that had high pitch in the first syllable. However, no preference was found when the familiarization stream alternated in duration. It is proposed that these perceptual biases help infants and adults find linguistic units in the continuous speech stream.While the bias for grouping based on pitch appears early in development, biases for durational grouping might rely on more extensive linguistic experience.


Mental Fatigue and Sport-Specific Psychomotor Performance: A Systematic Review.
Mental fatigue (MF) is a psychobiological state that impairs endurance performance in healthy athletes. Recently, multiple studies indicated that MF could also impair sport-specific psychomotor performance (SSPP). Nevertheless, a systematic overview detailing the effects of MF on SSPP is currently lacking. The objective of this study is to collate relevant literature and examine the effect of MF on SSPP. A secondary aim was to create an overview of the potential subjective and physiological factors underlying this MF effect. PubMed (MEDLINE), Web of Science, PsycINFO and SPORTDiscus were searched (5th of November 2020). Studies were eligible when study outcomes encompassed any form of SSPP skill in a sport-specific context, the intervention was targeted to induce MF, and the population included healthy individuals. The presence of a manipulation check, to indicate the successful induction of MF, was obligatory for inclusion. Secondary outcomes were all outcomes (either physiological or psychological) that could explain the underlying mechanisms of the effect of MF on SSPP. In total, 21 papers were included. MF was successfully induced in all but two studies, which were excluded from further analysis. MF negatively impacts a myriad of SSPP outcomes, including decision-making, reaction time and accuracy outcomes. No changes in physiological outcomes, that could underlie the effect of MF, were reported. Subjectively, only ratings of perceived of exertion increased due to MF in some studies. Overall, the selected papers indicated that MF negatively affects SSPP. Research that assesses brain function, while evaluating the effect of MF on SSPP is essential to create further insight.


Negative differential resistance in monolayer WTe2 tunneling transistors.
We report theoretical investigations of quantum transport in monolayer transition metal dichalcogenide (TMDC) tunneling field effect transistors (TFETs). Due to the specific electronic structure of TMDC WTe(2), a transmission valley is found in the conduction band (CB). For a proper choice of the doping, gate and supply voltages the WTe(2) TFET can produce a giant negative differential resistance (NDR) with a peak to valley ratio as large as 10(3). The mechanism of NDR is identified to be due to a transport-mode bottleneck, i.e., the band to band tunneling from the valence band of the source is partially blocked by a transmission valley of the CB of the drain. More generally, our calculations show that electronic structures of at least six TMDC materials possess the transmission valley.


The Effect of Simulation Wars on EM Residents' Global Rating Scales and In-Training Examination Scores.
The Accreditation Council for Graduate Medical Education (ACGME) restricted the duty hours for residents training in 2004. With less time to learn medicine, residents may not develop the clinical reasoning skills needed. Simulation can provide a remedy for this lack of time by allowing residents to practice skills and develop clinical reasoning in a simulated environment. Simulation Wars (SimWars), a clinical reasoning simulation has been shown to improve clinical reasoning skills. The purpose of the study was to investigate the effect of Simulation Wars on In-Training Examination (ITE) Scores and Global Rating Scale (GRS) Scores in Emergency Medicine (EM) residents. The Quasi-Experimental design was used in this retrospective study. The main comparison was between historical controls, the residents who did not participate in the Simulation Wars, and the intervention group comprised of residents who participated in the SimWars. A total of 127 residents were participants in this study including 70 from the intervention and 57 from the historical control group. There were no significant differences found in GRS scores between both groups except for Communication and Professionalism (p<0.001). No overall improvement in ITE scores for the control group and for the intervention group was found. Furthermore, within the intervention group, while comparing those residents who participated in certain subcategories of SimWars and those who did not, there was a significant improvement in ITE scores in the subcategories of Thoracic Disorders, Abdominal/Gastrointestinal, Trauma Disorders and OBGyn. SimWars in the subcategories of Abdominal/Gastrointestinal, Thoracic, OBGyn and Trauma were found to be associated with improved ITE scores in those subcategories. Since Emergency Medicine utilizes extensive clinical reasoning skills, SimWars may provide better educational opportunities for EM residents.


Why the inflammatory response is important to the cardiac surgical patient.
Although the systemic inflammatory response is recognized to contribute to patient morbidity and mortality after cardiopulmonary bypass, specific mechanisms linking cause and effect--linking a specific mediator with a defined adverse outcome--are lacking. The problem is partly because of the rarity of hard endpoints (stroke, myocardial injury, renal failure requiring dialysis), which studies are not always powered to measure, and partly one of definition; "systemic inflammatory response" wrongly suggests that the problem is confined to inflammation, whereas, in fact, it is characterized by systemic disturbances to a number of the body's natural defenses against injury and infection: fibrinolysis, coagulation, complement activation, immune cell activation, and oxidative stress in addition to inflammation. A better definition would be to think in terms of a "systemic host response" to surgery. End-organ injury results from the interplay of activated host defense systems with regional vessel wall injury, either because of physical trauma to the vein graft or ischemia/ reperfusion injury to susceptible vascular beds. Improved patient outcomes are going to take a concerted team effort to achieve, from the point of atraumatic vein harvest, to improved biocompatibility and shear resistance of circuits, monitoring, and minimizing of ischemia to organs, minimal cross-clamping trauma, optimized blood management, and combinatorial drug strategies. Surrogate endpoints for major organ dysfunction will play an important role to make sense of multiple interventions by the cardiac surgical team and to monitor continuous improvement to patient outcomes.


25-Hydroxyvitamin D, IL-31, and IL-33 in children with allergic disease of the airways.
Low vitamin D is involved in allergic asthma and rhinitis. IL-31 and IL-33 correlate with Th2-associated cytokines in allergic disease. We investigated whether low vitamin D is linked with circulating IL-31 and IL-33 in children with allergic disease of the airways. 25-Hydroxyvitamin D [25(OH) Vit D], IL-31, and IL-33 plasma levels were measured in 28 controls (HC), 11 allergic rhinitis (AR) patients, and 35 allergic asthma with rhinitis (AAR) patients. We found significant lower levels of 25(OH) Vit D in AR and in AAR than in HC. IL-31 and IL-33 plasma levels significantly increased in AAR than HC. IL-31 and IL-33 positively correlated in AR and AAR. 25(OH) Vit D deficient AAR had higher levels of blood eosinophils, exacerbations, disease duration, and total IgE than patients with insufficient or sufficient 25(OH) Vit D. In AAR 25(OH) Vit D levels inversely correlated with total allergen sIgE score and total atopy index. IL-31 and IL-33 did not correlate with 25(OH) Vit D in AR and AAR. In conclusion, low levels of 25(OH) Vit D might represent a risk factor for the development of concomitant asthma and rhinitis in children with allergic disease of the airways independently of IL-31/IL-33 Th2 activity.


Analysis of white spot syndrome virus envelope protein complexome by two-dimensional blue native/SDS PAGE combined with mass spectrometry.
White spot syndrome virus (WSSV) is a large enveloped virus, but the organization of its envelope proteins remains largely unknown. In the present study, we used blue native polyacrylamide gel electrophoresis (BN-PAGE) and SDS-PAGE in combination with mass spectrometry to analyze the envelope protein complexome of WSSV. Our results show that the viral envelope consists of multi-protein complexes (MPCs). Within them, the envelope protein VP19 exists as a homotrimer, while another major envelope protein, VP28, mainly exists as a homotetramer. The most notable feature is that the majority of MPCs include VP26 and VP24, suggesting that these two proteins might serve as hub proteins to recruit low-abundance proteins to MPCs and play crucial roles in the process of protein complex formation. Furthermore, we found significant evidence for interactions between several low-abundance proteins, such as VP52B/VP38/VP33 and VP12/VP150. The result of this study may promote the further research on WSSV envelope assembly.


Compensatory periplasmic nitrate reductase activity supports anaerobic growth of Pseudomonas aeruginosa PAO1 in the absence of membrane nitrate reductase.
Nitrate serves as a terminal electron acceptor under anaerobic conditions in Pseudomonas aeruginosa. Reduction of nitrate to nitrite generates a transmembrane proton motive force allowing ATP synthesis and anaerobic growth. The inner membrane-bound nitrate reductase NarGHI is encoded within the narK1K2GHJI operon, and the periplasmic nitrate reductase NapAB is encoded within the napEFDABC operon. The roles of the 2 dissimilatory nitrate reductases in anaerobic growth, and the regulation of their expressions, were examined by use of a set of deletion mutants in P. aeruginosa PAO1. NarGHI mutants were unable to grow anaerobically, but plate cultures remained viable up to 120 h. In contrast, the nitrate sensor-response regulator mutant DeltanarXL displayed growth arrest initially, but resumed growth after 72 h and reached the early stationary phase in liquid culture after 120 h. Genetic, transcriptional, and biochemical studies demonstrated that anaerobic growth recovery by the NarXL mutant was the result of NapAB periplasmic nitrate reductase expression. A novel transcriptional start site for napEFDABC expression was identified in the NarXL mutant grown anaerobically. Furthermore, mutagenesis of a consensus NarL-binding site monomer upstream of the novel transcriptional start site restored anaerobic growth recovery in the NarXL mutant. The data suggest that during anaerobic growth of wild-type P. aeruginosa PAO1, the nitrate response regulator NarL directly represses expression of periplasmic nitrate reductase, while inducing maximal expression of membrane nitrate reductase.


Microfluidic paper-based analytical devices fabricated by low-cost photolithography and embossing of Parafilm®.
Microfluidic paper-based analytical devices (μPADs) attract tremendous attention as an economical tool for in-field diagnosis, food safety and environmental monitoring. We innovatively fabricated 2D and 3D μPADs by photolithography-patterning microchannels on a Parafilm® and subsequently embossing them to paper. This truly low-cost, wax printer and cutter plotter independent approach offers the opportunity for researchers from resource-limited laboratories to work on paper-based analytical devices.


The pleckstrin homology domain in the SKAP55 adapter protein defines the ability of the adapter protein ADAP to regulate integrin function and NF-kappaB activation.
Adhesion and degranulation promoting adapter protein (ADAP) is a multifunctional hematopoietic adapter protein that regulates TCR-dependent increases in both integrin function and activation of the NF-κB transcription factor. Activation of integrin function requires both ADAP and the ADAP-associated adapter Src kinase-associated phosphoprotein of 55 kDa (SKAP55). In contrast, ADAP-mediated regulation of NF-κB involves distinct binding sites in ADAP that promote the inducible association of ADAP, but not SKAP55, with the CARMA1 adapter and the TAK1 kinase. This suggests that the presence or absence of associated SKAP55 defines functionally distinct pools of ADAP. To test this hypothesis, we developed a novel SKAP-ADAP chimeric fusion protein and demonstrated that physical association of ADAP with SKAP55 is both sufficient and necessary for the rescue of integrin function in ADAP-deficient T cells. Similar to wild-type ADAP, the SKAP-ADAP chimera associated with the LFA-1 integrin after TCR stimulation. Although the SKAP-ADAP chimera contains the CARMA1 and TAK1 binding sequences from ADAP, expression of the chimera does not restore NF-κB signaling in ADAP(-/-) T cells. A single point mutation in the pleckstrin homology domain of SKAP55 (R131M) blocks the ability of the SKAP-ADAP chimera to restore integrin function and to associate with LFA-1. However, the R131M mutant was now able to restore NF-κB signaling in ADAP-deficient T cells. We conclude that integrin regulation by ADAP involves the recruitment of ADAP to LFA-1 integrin complexes by the pleckstrin homology domain of SKAP55, and this recruitment restricts the ability of ADAP to interact with the NF-κB signalosome and regulate NF-κB activation.


A versatile and practical method for regioselective synthesis of polysubstituted furanonaphthoquinones.
An efficient and attractive synthesis of a series of novel poly-functionalized phosphorus zwitterions was achieved via three-component reactions of the corresponding functional nucleophiles, aldehydes, and Bu(3)P in the presence of acidic promoter. These polysubstituted zwitterions could regioselectively undergo further transformations to synthetically important furanonaphthoquinones and related products via the intramolecular Wittig reaction. These methods could have potential application in synthetic and pharmaceutical chemistry for its facilitation and easily accessible commercial materials.


Immunohistochemical analysis of DNA synthesis during chronic stimulation with isoproterenol in mouse submandibular gland.
We investigated the numbers of DNA-synthesizing cells in mouse submandibular glands (SMGs) during chronic isoproterenol (IPR) administration, using an immunohistochemical method with anti-bromodeoxyuridine (BrdU) antibodies. Adult and immature female mice were injected with IPR daily or every other day up to a total of seven injections. Whereas hypertrophic enlargement of acinar cells continued with repeated IPR injections, induction of DNA synthesis was transient. With one IPR injection, more than 50% of the acinar cells were induced to synthesize DNA, but after five daily injections the numbers of cells in the DNA synthetic phase decreased and returned to control values. This pattern was basically the same for immature mice and for adult mice stimulated by IPR every other day. The responsiveness of the SMGs of immature mice to IPR was somewhat lower than that of adults. Prolongation of the interval between injections of IPR, compared to daily injections, did not cause sustained induction of DNA synthesis. These data suggest that the hypertrophic enlargement and the induction of DNA synthesis by IPR are mediated through different intracellular pathways after the stimulation of beta-adrenergic receptors, and that the regulatory mechanism for the induction of DNA synthesis is complex.


Effects of PCSK9 genetic variants on plasma LDL cholesterol levels and risk of premature myocardial infarction in the Italian population.
The R46L variant in the proprotein-convertase subtilisin-kexin type 9 (PCSK9) gene was associated with reduced levels of LDL and total cholesterol and with a lower risk of coronary artery disease. We investigated the association of R46L with myocardial infarction (MI) in 1,880 Italian patients with premature MI and 1,880 controls. A trend toward a protective effect of the L46 allele was observed [odds ratio (OR) = 0.75, 95% confidence interval (CI) = 0.49-1.13; P = 0.17], although the association with MI was not significant. This is probably due to the combined effect of the low frequency of R46L among Italians and of the young age of the analyzed cohort for whom the impact of coronary atherosclerosis is less important. This hypothesis was indirectly confirmed by the significant association found after including 1,056 additional older controls (OR = 0.67, 95% CI = 0.46-0.97; P = 0.036). LDL cholesterol was significantly lower in L46 carriers (116.2 ± 34.7 mg/dl) than in noncarriers (137.4 ± 47.3 mg/dl; P = 0.00022); a similar reduction was observed for total cholesterol (191.7 ± 37.7 vs. 211.7 ± 49 mg/dl; P = 0.00019). Analysis of 23 additional polymorphisms in the PCSK9 region identified another single nucleotide polymorphism (SNP) (rs11206510) associated with cholesterol levels. We confirmed that the L46 allele not only decreases LDL cholesterol but also protects against MI. Moreover, we replicated the association of total and LDL cholesterol with the SNP rs11206510.


Psychometric properties of the Family Accommodation Scale for Obsessive-Compulsive Disorder-Patient Version.
In obsessive-compulsive disorder (OCD), family accommodation is a frequently occurring phenomenon that has been linked to attenuated treatment response, increased obsessive-compulsive symptom severity, and lower levels of functioning. No patient-report version of family accommodation exists, with available measures relying on relatives as informants. However, adult patients with OCD often present to clinical services alone, frequently making it impractical to obtain information from these informants. Consequently, a standardized patient-reported measure of family accommodation proves salient in clinical practice. The present study examined the psychometric properties of the Family Accommodation Scale for Obsessive-Compulsive Disorder-Patient Version (FAS-PV). Sixty-one adults with OCD were administered clinician-rated measures of OCD symptom severity and self-report questionnaires examining functional impairment, family functioning, and emotional/behavioral difficulties. Fifty-four relatives completed self-report measures assessing family accommodation and family functioning. The majority of the adult OCD participants (89%) endorsed at least 1 type of accommodating behavior in the previous week. The FAS-PV total score demonstrated good internal consistency and test-retest reliability. Convergent validity was evidenced by strong associations with scores on another measure of family accommodation, OCD symptom severity, OCD-related family functioning, anxiety, and functional impairment. Divergent validity was supported through nonsignificant correlations with depressive symptoms and impulsivity. The FAS-PV did not significantly differ from the relative-reported measure of family accommodation in terms of the internal consistency or mean of the total scores. Ultimately, the FAS-PV scores demonstrated sound psychometric properties and validity in assessing family accommodation from the patient's perspective, encouraging its use in research and clinical practice.


Extracorporeal membrane oxygenation for respiratory failure in adults.
This article reviews case series and trials that evaluated extracorporeal membrane oxygenation (ECMO) for respiratory failure and describes patient and circuit management in the modern era of ECMO support. In recent years, pivotal progress has been made in the conception and construction of ECMO circuits. They are now simpler, safer, require less anticoagulation and are associated with fewer bleeding complications. The encouraging results of the efficacy and economic assessment of conventional ventilatory support versus ECMO for severe adult respiratory failure (CESAR) trial performed in the United Kingdom and good outcomes of patients who received ECMO as rescue therapy during the recent H1N1 influenza pandemic, in which the latest generation of ECMO technology was used, reignited interest in ECMO for severe acute respiratory distress syndrome (ARDS). The latest generation of ECMO systems is more biocompatible, better performing and longer lasting. Although recent studies suggested that veno-venous ECMO might improve the outcomes of patients with ARDS, indications for ECMO use remain uncertain. Future trials of ECMO for severe ARDS should strictly control for standard-of-care mechanical ventilation strategies in the control group and early transportation on ECMO for patients in the intervention arm.


MapCHECK used for rotational IMRT measurements: step-and-shoot, TomoTherapy, RapidArc.
To measure patient-specific-QA dose distributions with a 2D array of diodes, the MapCHECK, for the dose delivered, with step-and-shoot and rotational IMRT. Two MapCHECKs were used that had different styles of diode connection. These MapCHECKs were used in their original manufactured configuration and in a modified configuration. The modification made in the clinic consists of filling air gaps with sheets of Lucite that had custom-machined slots for the diodes and by adding pieces of copper to offset the intrinsic asymmetry of the diodes. The MapCHECKs were housed in a tight-fitting phantom fabricated from solid water. Measurements were made on IMRT treatment plans delivered with Varian linear accelerators with step-and-shoot and RapidArc methods, and a TomoTherapy machine with helical delivery. Patient plans and QA plans were developed with the XiO, Eclipse, and TomoTherapy planning systems. All MapCHECK data were analyzed with its commercially available software. Kilovoltage CT imaging of the MapCHECK in its phantom has streak artifacts from high atomic number components. These artifacts must be corrected in order to obtain accurate calculation of dose. The original MapCHECK is found to have an angular dependence of +/- 20%. A modification of the MapCHECK has been made that reduces the angular dependence to +/- 2%. Proper compensation for attenuation by the treatment couch is also necessary for accurate results. The modified MapCHECK has been successfully used for doing patient-specific QA for IMRT treatments delivered with step-and-shoot, TomoTherapy, and RapidArc methods. Treatment plans that require large amounts of fluence from the 90 degrees and 270 degrees directions have significantly better QA measurements when made with modified MapCHECKs. When properly modified, the MapCHECK response becomes isotropic. It can then be used for patient-specific QA measurements for IMRT treatments delivered with step-and-shoot and rotational techniques such as helical Tomotherapy and RapidArc.


Serum liver enzyme and histopathologic changes in calves with chronic and chronic-delayed Senecio jacobaea toxicosis.
Progressive changes in serum enzyme activity and liver histologic features were monitored in calves fed tansy ragwort (Senecio jacobaea)-contaminated pellets. The experiments were designed to simulate natural intoxicant ingestion conditions in relationship to the dose and duration of exposure to the toxic plant to correlate early laboratory diagnostic changes with the natural progression of the disease, thereby facilitating early diagnosis and intervention by veterinary clinicians. Eight calves were fed tansy ragwort and 4 additional calves served as controls. In group 1, 4 calves were continuously fed dried tansy ragwort mixed in a pelleted feed at a 5% concentration by dry weight until terminal liver disease developed. Serum liver enzyme (alkaline phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase) activities were monitored at weekly intervals in these calves and in the 2 controls. In group 2, 4 calves were fed the same contaminated feed for only 60 days, with return to normal feed for the duration of the trial. Two additional calves served as controls. Their liver enzyme activities were monitored every other week in conjunction with percutaneous liver biopsies. All 8 calves fed tansy ragwort-contaminated pellets developed terminal hepatopathy in either a chronic pattern (n = 6) or a chronic-delayed pattern (n = 2), with the onset of a moribund state or sudden death at 11 to 17 weeks and 27 to 51 weeks, respectively. The calves were euthanatized when classic terminal signs of hepatic encephalopathy first became evident. The clinicopathologic patterns of chronic and chronic-delayed toxicoses were typical of over 5,000 cases of field tansy toxicosis diagnosed at the diagnostic laboratory. Serum glutamate dehydrogenase was the first enzyme to increase in most animals, with a short-term increase to peak values followed by a rapid return to normal. This enzyme change was followed by increases in alkaline phosphatase and gamma-glutamyltransferase. Serum enzyme changes preceded development of recognizable histologic lesions. Vacuolar changes in hepatocyte nuclei, biliary hyperplasia, and fibrosis sequentially developed in liver biopsy specimens from each animal, whereas megalocytosis was not a predominant feature until necropsy. On the basis of our findings, we suggest that the optimal tests for diagnosis of pyrrolizidine alkaloid intoxication should consist of liver biopsy and determination of concurrent serum liver-enzyme activities.


A pair of 3D enantiotopic zinc(ii) complexes based on two asymmetric achiral ligands.
Enantiomorphism and enantioselectivity are critical in biology and many other applications. Herein, we report two 3D chiral MOFs {[Zn6(MIDPPA)3(1,2,4-btc)3(NO2)3(H2O)3](H2O)7}n (1L and 1R) based on achiral ligands with high enantiomeric excess and a novel topological type. The internal mechanism of spontaneous chiral symmetry breaking, during the crystallization of chiral MOFs based on achiral ligands, is elucidated for the first time from both structural and theoretical aspects. Hydrogen bonds are found to play a key role in the spontaneous symmetry breaking of chiral MOFs. Also, DFT calculations support our findings from the aspects of total energies and HOMO-LUMO energy gaps. Both 1L and 1R, exhibiting green fluorescence, present a non-centrosymmetric polar packing arrangement, resulting in good ferroelectric behaviors and second-order nonlinear optical effects.


The e-Psychonauts' 'Spiced' World; Assessment of the Synthetic Cannabinoids' Information Available Online.
A wide range of novel psychoactive substances (NPS) is regularly searched and discussed online by web-based drug enthusiasts (i.e. the e-psychonauts). Among NPS, the range of synthetic cannabinoids (SC; 'Spice') currently represents a challenge for governments and clinicians. Using a web crawler (i.e. the NPS.Finder®), the present study aimed at assessing psychonauts' fora/platforms to better understand the online mentions of SC. The open-web crawling/navigating software identified here some 1,103 synthetic cannabinoids. Of these, 863 molecules were not listed in either the international or the European NPS databases. A web crawling approach helped here in identifying a large range of unknown SC likely to possess a misuse potential. Most of these novel/emerging molecules are still relatively unknown. This is a reason for concern; each of these analogues potentially presents different toxicodynamic profiles and there is a lack of docking, preclinical, and clinical observations. Strengthening multidisciplinary collaboration between clinicians and bioinformatics may prove useful in better assessing SC-associated public health risks.


Potential effects of aggressive decongestion during the treatment of decompensated heart failure on renal function and survival.
Overly aggressive diuresis leading to intravascular volume depletion has been proposed as a cause for worsening renal function during the treatment of decompensated heart failure. If diuresis occurs at a rate greater than extravascular fluid can refill the intravascular space, the concentration of such intravascular substances as hemoglobin and plasma proteins increases. We hypothesized that hemoconcentration would be associated with worsening renal function and possibly would provide insight into the relationship between aggressive decongestion and outcomes. Subjects in the Evaluation Study of Congestive Heart Failure and Pulmonary Artery Catheterization Effectiveness trial limited data set with a baseline/discharge pair of hematocrit, albumin, or total protein values were included (336 patients). Baseline-to-discharge increases in these parameters were evaluated, and patients with >or=2 in the top tertile were considered to have evidence of hemoconcentration. The group experiencing hemoconcentration received higher doses of loop diuretics, lost more weight/fluid, and had greater reductions in filling pressures (P<0.05 for all). Hemoconcentration was strongly associated with worsening renal function (odds ratio, 5.3; P<0.001), whereas changes in right atrial pressure (P=0.36) and pulmonary capillary wedge pressure (P=0.53) were not. Patients with hemoconcentration had significantly lower 180-day mortality (hazard ratio, 0.31; P=0.013). This relationship persisted after adjustment for baseline characteristics (hazard ratio, 0.16; P=0.001). Hemoconcentration is significantly associated with measures of aggressive fluid removal and deterioration in renal function. Despite this relationship, hemoconcentration is associated with substantially improved survival. These observations raise the question of whether aggressive decongestion, even in the setting of worsening renal function, can positively affect survival.