Mammalian cell metabolomics: experimental design and sample preparation.
Metabolomics represents the global assessment of metabolites in a biological sample and reports the closest information to the phenotype of the biological system under study. Mammalian cell metabolomics has emerged as a promising tool with potential applications in many biotechnology and research areas. Metabolomics workflow includes experimental design, sampling, sample processing, metabolite analysis, and data processing. Given their influence on metabolite content and biological interpretation of data, a good experimental design and the appropriate choice of a sample processing method are prerequisites for success in any metabolomic study. The use of mammalian cells in the metabolomics field involves harder sample processing methods, including metabolism quenching and metabolite extraction, as compared to the use of body fluids, although such critical issues are frequently overlooked. This review aims to overview the common experimental procedures used in mammalian cell metabolomics based on mass spectrometry, by placing special emphasis on discussing sample preparation approaches, although other aspects, such as cell metabolomics applications, culture systems, cellular models, analytical platforms, and data analysis, are also briefly covered. This review intends to be a helpful tool to assist researchers in addressing decisions when planning a metabolomics study involving the use of mammalian cells.


Ubiquitin ligase ATL31 functions in leaf senescence in response to the balance between atmospheric CO2 and nitrogen availability in Arabidopsis.
Carbon (C) and nitrogen (N) are essential elements for metabolism, and their availability, called the C/N balance, must be tightly coordinated for optimal growth in plants. Previously, we have identified the ubiquitin ligase CNI1/ATL31 as a novel C/N regulator by screening plants grown on C/N stress medium containing excess sugar and limited N. To elucidate further the effect of C/N balance on plant growth and to determine the physiological function of ATL31, we performed C/N response analysis using an atmospheric CO2 manipulation system. Under conditions of elevated CO2 and sufficient N, plant biomass and total sugar and starch dramatically increased. In contrast, elevated CO2 with limited N did not increase plant biomass but promoted leaf chlorosis, with anthocyanin accumulation and increased senescence-associated gene expression. Similar results were obtained with plants grown in medium containing excess sugar and limited N, suggesting that disruption of the C/N balance affects senescence progression. In ATL31-overexpressing plants, promotion of senescence under disrupted CO2/N conditions was repressed, whereas in the loss-of-function mutant it was enhanced. The ATL31 gene was transcriptionally up-regulated under N deficiency and in senescent leaves, and ATL31 expression was highly correlated with WRKY53 expression, a key regulator of senescence. Furthermore, transient protoplast analysis implicated the direct activation of ATL31 expression by WRKY53, which was in accordance with the results of WRKY53 overexpression experiments. Together, these results demonstrate the importance of C/N balance in leaf senescence and the involvement of ubiquitin ligase ATL31 in the process of senescence in Arabidopsis.


Exhaled Nitric Oxide in Systemic Sclerosis Lung Disease.
Background. Exhaled nitric oxide (eNO) is a potential biomarker to distinguish systemic sclerosis (SSc) associated pulmonary arterial hypertension (PAH) and interstitial lung disease (ILD). We evaluated the discriminative validity, feasibility, methods of eNO measurement, and magnitude of differences across lung diseases, disease-subsets (SSc, systemic lupus erythematosus), and healthy-controls. Methods. Consecutive subjects in the UHN Pulmonary Hypertension Programme were recruited. Exhaled nitric oxide was measured at 50 mL/s intervals using chemiluminescent detection. Alveolar and conducting airway NO were partitioned using a two-compartment model of axial diffusion (CMAD) and the trumpet model of axial diffusion (TMAD). Results. Sixty subjects were evaluated. Using the CMAD model, control subjects had lower median (IQR) alveolar NO than all PAH subjects (2.0 (1.5, 2.5) versus 3.14 ppb (2.3, 4.0), p = 0.008). SSc-ILD had significantly lower median conducting airway NO compared to controls (1009.5 versus 1342.1 ml⁎ppb/s, p = 0.04). SSc-PAH had increased median (IQR) alveolar NO compared to controls (3.3 (3.0, 5.7) versus 2.0 ppb (1.5, 2.5), p = 0.01). SSc-PAH conducting airway NO inversely correlated with DLCO (r -0.88 (95% CI -0.99, -0.26)). Conclusion. We have demonstrated feasibility, identified that CMAD modeling is preferred in SSc, and reported the magnitude of differences across cases and controls. Our data supports discriminative validity of eNO in SSc lung disease.


Side effects of antibiotics on the intestinal microflora by PCR-DGGE.
Antibiotics are recommended for use in the treatment of infectious diseases, but the side effects have not been thoroughly investigated, especially on the intestinal tract. Fluoroquinolones, macrolides and β-lactams were tested the side effects on the intestinal microbflora of mice in this study. The similarity and sequence analysis of the dominant bands for different drug administration periods were carried out by denaturing gradient gel electrophoresis (DGGE) profiles. The total amount of 16S rRNA gene increased after drug administration, the bacterial composition and structure could be divided into different clusters for different drug administration periods distinctly. This study revealed a significant change of bacterial composition of microflora from intestinal tract as antibiotics were applied to tested mice.


Myelinated and nonmyelinated nerves: comparison of proton MR properties.
The magnetic resonance (MR) relaxation rates of protons were compared in the myelinated and nonmyelinated nerves of the garfish. The long, large olfactory nerve of the garfish, as an easily accessible source of nonmyelinated axons, is uniquely suited for such a comparison. The T1 and T2 measurements revealed distinct and consistent differences between nonmyelinated olfactory nerves and myelinated optic and oculomotor nerves. Comparisons between water content, lipid content, and relaxation rates indicated that the differences in MR properties represent complex differences in the distribution and physical environment of the constituent lipid and water protons.


Interpretation of Dihydrorhodamine-1,2,3 Flow Cytometry in Chronic Granulomatous Disease: an Atypical Exemplar.
This is a functional characterization of a novel CYBA variant associated with normal DHR flow cytometry. Chronic granulomatous disease (CGD) is an inborn error of immunity characterized by recurrent bacterial and fungal infections and dysregulated inflammatory responses due to defective phagocytic cell function leading to the formation of granulomas. CGD patients have pathogenic variants in any of the five components of the phagocytic NADPH oxidase, which transfers electrons through the phagosomal membrane and produces superoxide upon bacterial uptake. Here, we report a pediatric female patient with a novel homozygous missense variant (c.293C > T, p.(Ser98Leu)) in CYBA, encoding the p22phox protein, associated with autosomal recessive CGD. The patient presented with severe recurrent pneumonia. Specific pathogens identified included Burkholderia and Serratia species suggesting neutrophil functional abnormalities; however, the dihydrorhodamine-1,2,3 (DHR) flow cytometric and cytochrome c reduction assays for neutrophil respiratory burst fell within the low side of the normal range. Western blot and flow cytometric analysis of individual NADPH oxidase components revealed reduced levels of p22phox and gp91phoxphox proteins. The pathological consequence of the p.Ser98Leu variant was further evaluated in heterologous expression systems, which confirmed reduced p22phox protein stability and oxidase activity. Although this patient did not exhibit all the classic features of CGD, such as granulomas and skin infections, she had recurrent pneumonias with oxidant-sensitive pathognomonic organisms, resulting in appropriate targeted CGD testing. This case emphasizes the need to contextually interpret laboratory data, especially using clinical findings to direct additional assessments including genetic analysis.


An enzymatic activity in bovine brain that catalyzes the reversal of the C-terminal methyl esterification of protein phosphatase 2A.
A novel protein methyltransferase has been recently described that catalyzes the esterification of the C-terminal leucine residue of the catalytic subunit of protein phosphatase 2A in a variety of eucaryotic cells. This reaction can potentially modulate the phosphatase's activity, subunit interactions, or interactions with specific phosphoprotein substrates. We present evidence here that the methylation reaction is reversible and that an enzymatic activity is present in bovine brain cytosol that catalyzes the hydrolysis of the methyl ester. We show that this activity is sensitive to inhibition by the serine-hydrolase inhibitor phenylmethanesulfonyl fluoride but is not affected by the small molecule substrate analog N-acetyl-L-leucine methyl ester. These results suggest that protein methylation and demethylation reactions can be utilized in eucaryotic cells to modulate enzyme activity in a parallel fashion to protein phosphorylation and dephosphorylation reactions.


[Gene Replacement Therapy for Inherited Retinal Dystrophies].
Characteristics of inherited retinal dystrophies include deficiencies in light perception and nervous conduction within the retina, leading to reduced vision or even blindness. In this context, the loss of function of photoreceptor-specific genes causes a variety of clinically and aetiologically distinct syndromes - each of them belonging to the group of rare diseases. With a prevalence of 1 in 2500, however, inherited retinal diseases are clinically significant and important - especially since these diseases lead to restrictions of a patient's fitness for work and overall quality of life. More than 250 genetic mutations causing the various types of inherited retinal dystrophies have been identified by now (https://sph.uth.tmc.edu/Retnet). In recent years, preclinical research on suitable animal models has yielded important progress in the understanding of the mutations underlying the pathological and molecular biological processes of these diseases. These findings have led to the development of novel and innovative therapeutic strategies for the treatment of inherited retinal dysfunctions, which are still incurable. Meanwhile, many of the successful preclinical studies have led to translational research projects aiming to find treatment options for human patients. However, some preliminary results of these human translational studies indicate the need to optimise and refine the underlying therapeutic concepts.


Conflicting pathology reports: a diagnostic dilemma.
The differential diagnosis of a brain lesion with two discordant pathology reports includes the presence of collision tumor, metaplastic changes, and labeling errors that occurred during the processing of the specimen. The authors present a case in which the first brain biopsy from a 47-year-old patient with a history of heavy smoking was compatible with metastatic small cell carcinoma, and the second biopsy taken during decompression craniotomy 3 weeks later was compatible with WHO Grade IV glioblastoma. Using short tandem repeat (STR) analysis of the two specimens and nontumor-derived patient DNA, the authors found that the two specimens did not belong to the same individual. The authors conclude that allele imbalance or loss of heterozygosity detected by STR analysis is a reliable and valuable diagnostic tool for clarifying discrepancies in discordant pathology reports.


Novel, selective CDK9 inhibitors for the treatment of HIV infection.
Cyclin Dependent Kinases (CDKs) are important regulators of cell cycle and gene expression. Since an up-to-date review about the pharmacological inhibitors of CDK family (CDK1-10) is not available; therefore in the present paper we briefly summarize the most relevant inhibitors and point out the low number of selective inhibitors. Among CDKs, CDK9 is a validated pathological target in HIV infection, inflammation and cardiac hypertrophy; however selective CDK9 inhibitors are still not available. We present a selective inhibitor family of CDK9 based on the 4-phenylamino-6- phenylpyrimidine nucleus. We show a convenient synthetic method to prepare a useful intermediate and its derivatisation resulting in novel compounds. The CDK9 inhibitory activity of the derivatives was measured in specific kinase assay and the CDK inhibitory profile of the best ones (IC(50) < 100 nM) was determined. The most selective compounds had high selectivity over CDK1, 2, 3, 5, 6, 7 and showed at least one order of magnitude higher inhibitory activity over CDK4 inhibition. The most selective molecules were examined in cytotoxicity assays and their ability to inhibit HIV-1 replication was determined in cellular assays.


Machine learning to parse breast pathology reports in Chinese.
Large structured databases of pathology findings are valuable in deriving new clinical insights. However, they are labor intensive to create and generally require manual annotation. There has been some work in the bioinformatics community to support automating this work via machine learning in English. Our contribution is to provide an automated approach to construct such structured databases in Chinese, and to set the stage for extraction from other languages. We collected 2104 de-identified Chinese benign and malignant breast pathology reports from Hunan Cancer Hospital. Physicians with native Chinese proficiency reviewed the reports and annotated a variety of binary and numerical pathologic entities. After excluding 78 cases with a bilateral lesion in the same report, 1216 cases were used as a training set for the algorithm, which was then refined by 405 development cases. The Natural language processing algorithm was tested by using the remaining 405 cases to evaluate the machine learning outcome. The model was used to extract 13 binary entities and 8 numerical entities. When compared to physicians with native Chinese proficiency, the model showed a per-entity accuracy from 91 to 100% for all common diagnoses on the test set. The overall accuracy of binary entities was 98% and of numerical entities was 95%. In a per-report evaluation for binary entities with more than 100 training cases, 85% of all the testing reports were completely correct and 11% had an error in 1 out of 22 entities. We have demonstrated that Chinese breast pathology reports can be automatically parsed into structured data using standard machine learning approaches. The results of our study demonstrate that techniques effective in parsing English reports can be scaled to other languages.


AM67, a secretory component of the guinea pig sperm acrosomal matrix, is related to mouse sperm protein sp56 and the complement component 4-binding proteins.
The guinea pig sperm acrosomal matrix is the dense core of the acrosome and is likely to be important in acrosome biogenesis and fertilization. Isolated acrosomal matrices are composed of a limited number of major bands when analyzed by SDS-polyacrylamide gel electrophoresis, among which is a Mr 67,000 protein that we have termed AM67. Indirect immunofluorescence demonstrated that AM67 is localized to the apical segment of the cauda epididymal sperm acrosome. Immunoelectron microscopy further refined the localization of AM67 to the M1 (dorsal bulge) domain within the acrosome. Using a polymerase chain reaction product based upon tryptic peptide sequences from AM67, a lambdagt11 guinea pig testis cDNA library was screened to yield two cDNA clones that encode the AM67 peptides. Northern analysis revealed that AM67 is transcribed as a 1. 9-kilobase testis-specific mRNA. The complete AM67 sequence encodes a prepropolypeptide of 533 amino acids with a calculated Mr of 59, 768. Following cleavage of a probable signal sequence, the polypeptide was predicted to have a Mr of 56,851 and seven consensus sites for asparagine-linked glycosylation. The deduced amino acid sequence of AM67 is most similar to those of the mouse sperm protein sp56 and the alpha-subunits of complement component 4-binding proteins from various mammalian species. Although mouse sp56 has been reported to be a cell-surface receptor for the murine zona pellucida glycoprotein ZP3, standard immunoelectron microscopy using the anti-sp56 monoclonal antibody 7C5 detected sp56 within the mouse sperm acrosome, but failed to detect sp56 on the surface of acrosome-intact mouse sperm. Furthermore, acrosomal labeling was detected in mouse sperm prepared for immunofluorescence using paraformaldehyde fixation, but was not observed with live unfixed sperm. Thus, the finding that sp56 is present within the acrosome provides further support that sp56 and AM67 are orthologues and suggests that sp56 may function in acrosomal matrix-zona pellucida interactions during and immediately following the acrosome reaction in the mouse.


Scar endometrioma: awareness and prevention.
This article presents two cases of abdominal wall endometrioma. Endometrioma develops after surgery due to transportation and subsequent implantation of endometrial cells. Abdominal wall endometrioma can cause pain, anxiety, and morbidity. Therefore implementation of preventative measures during surgery is paramount to avoid extraneous surgery and hospital expenses. Diagnoses are rarely established prior to surgery, but surgeons often suspect the syndrome in women who have a history of a mass associated with cyclical pain. Radiological examination is useful to exclude hernia and metastatic malignancy, and to delineate the extent of the mass. Wide excision of endometrioma is usually curative.


Thyroid Hormone Receptor Alpha Deletion in ApoE-/- Mice Alters the Arterial Renin-Angiotensin System and Vascular Smooth Muscular Cell Cholesterol Metabolism.
Thyroid hormone (TH) regulates gene transcription by binding to TH receptors (TRs). TRs regulate the genes of lipid metabolism and the renin-angiotensin system (RAS). We examined the effect of TRα deletion in ApoE-/- mice (DKO mice) on the following: (i) the expression of genes controlling cholesterol metabolism and tissue (t)RAS in the liver and aorta and (ii) the expression of these genes and the regulation of cholesterol content in cultured vascular smooth muscle cells (VSMCs). TRα deletion in ApoE-/- mice led to the repression of genes involved in the synthesis and influx of cholesterol in the liver. However, TRα deletion in the arterial wall suppressed the expression of genes involved in the esterification and excretion of cholesterol and enhanced the expression of angiotensinogen (AGT). The VSMCs of the ApoE-/- and DKO mice increased their cholesterol content during cholesterol loading, but failed to increase the expression of ATP-binding cassette transporter A1 (ABCA1). T3 addition partially corrected these abnormalities in the cells of the ApoE-/- mice but not those of the DKO mice. In conclusion, TRα deletion in ApoE-/- mice slightly increases the expression of tRAS in the aorta and aggravates the dysregulation of cholesterol content in the VSMCs.


Collaborative hand surgery clinical research without sharing individual patient data; proof of principle study.
High-quality research in hand surgery is increasingly important. A vital component is national and international multicenter collaborative research because of better generalizability and larger sample sizes. However, sharing patient data between centers can be hampered by regulations and privacy issues or reluctance to share patient data. Therefore, in this paper, we illustrate an approach for collaborative clinical research without sharing patient data while obtaining similar outcomes. To illustrate that this collaborative clinical research approach without sharing patient data leads to similar outcomes compared to aggregating all individual patient data in one database, we simulate an approach of performing meta-analyses on summary statistics of individual-center data. In the simulation, we compare the results to conventional analyses in an existing multicenter database of patients treated for Dupuytren's disease at three different centers with either limited fasciectomy (LF) or needle aponeurotomy (PNF). We share example data and all analysis code in a public GitHub Library. We found similar results for the meta-analysis approach without sharing individual patient data as in the conventional approach for 1) the proportion of patients treated for recurrences, 2) the Total MHQ score after both treatments, 3) the comparison of Total MHQ score after both treatments, and 4) the comparison of both treatments when correcting for confounders with regression analysis. CLINICAL SIGNIFICANCE: We illustrate how collaborative studies can be performed without sharing individual patient data while obtaining similar results as with conventional analyses. This approach can help speed up collaborative research without losing precision in outcome analysis.


Processing of the primer for plus strand DNA synthesis by human immunodeficiency virus 1 reverse transcriptase.
We have analyzed the processing of the RNA primer for (+) strand DNA synthesis by reverse transcriptase of the human immunodeficiency virus 1. To test for specific RNA cleavage and primer usage, we constructed a 99-base pair RNA-DNA hybrid containing the viral polypurine tract and flanking viral sequences. Although the RNase H activity of reverse transcriptase cleaves the RNA strand into multiple fragments, only two primers are extended in the presence of nucleoside triphosphates. The major RNA primer includes the entire polypurine tract except for the last adenosine and has the sequence 5'-UUUUAAAAGAAAAGGGGGG-3'. The minor primer has the same 3' end but is two nucleotides shorter. In a subsequent processing step reverse transcriptase releases the primer intact via a cleavage at the RNA-DNA junction. RNA cleavage, primer extension, and primer removal can take place in a single reaction. However, specificity does not require coupling of the three steps and is preserved in the individual reactions. The polypurine primer is generated and removed after its elongation in the absence of DNA synthesis. Furthermore, the polypurine primer is selected among the several RNA fragments available and extended by reverse transcriptase as well as by p51, a short form of reverse transcriptase lacking RNase H activity.


Predictors of axillary lymph node metastases (ALNM) in a Korean population with T1-2 breast carcinoma: triple negative breast cancer has a high incidence of ALNM irrespective of the tumor size.
We estimated the likelihood of breast cancer patients having axillary lymph node metastases (ALNM) based on a variety of clinical and pathologic factors. Three hundred sixty-one breast cancer patients without distant metastases and who underwent breast conserving surgery and axillary lymph node dissection (ALND) (level I and II) or modified radical mastectomy (MRM) were identified, and we retrospectively reviewed their pathology records and treatment charts. Positive axillary lymph nodes were detected in 104 patients for an overall incidence of 28.8%: 2 patients (5%) with T1a tumor, 5 (9.2%) with T1b tumor, 24 (21.8%) with T1c tumor and 73 (44.2%) with T2 tumor. On the multivariate analysis, an increased tumor size (adjusted OR=11.87, p=0.02), the presence of lymphovascular invasion (adjusted OR=7.41, p<0.01), a triple negative profile (ER/PR-, Her2-) (adjusted OR=2.09, p=0.04) and a palpable mass at the time of diagnosis (adjusted OR=2.31, p=0.03) were all significant independent factors for positive ALNM. In our study, the tumor size, the presence of lymphovascular invasion, a triple negative profile and a palpable mass were the independent predictive factors for ALNM. The tumor size was the strongest predictor of ALNM. Thus, the exact estimation of the extent of tumor is necessary for clinicians to optimize the patients' care. Patients with a triple negative profile have a high incidence of ALNM irrespective of the tumor size.


Prothrombotic and lipoprotein variables in patients attending a cardiovascular risk management clinic: response to ciprofibrate or lifestyle advice.
Lipid lowering drugs improve survival. However, intervention studies have focused on reducing serum total cholesterol and low density lipoprotein (LDL) concentrations and have not considered that levels of high density lipoprotein (HDL), triglycerides (TG), lipoprotein (a) and fibrinogen also predict risk and outcome. A retrospective survey of patients with primary dyslipidaemia attending a cardiovascular risk management clinic (set in a university hospital) was initiated to assess the effect of ciprofibrate, a drug with the potential to modify all these variables. Patients who received ciprofibrate (n = 72) were compared with 64 patients who only received lifestyle advice. Both groups had a similar age and gender distribution. The ciprofibrate group had a higher cardiovascular risk load but both groups shared several other characteristics. Fasting serum total cholesterol, LDL, HDL, TG, lipoprotein (a), glucose and plasma fibrinogen concentrations were measured at baseline and after 2-4 months. Ciprofibrate significantly improved total cholesterol, LDL, HDL, TG, lipoprotein (a) and fibrinogen. In contrast, lifestyle advice only significantly (but less markedly) reduced serum total cholesterol and TG concentrations. In a clinical setting, ciprofibrate has a broad spectrum of action on several predictors of vascular events. Although our study is not of a double blind randomized design it reflects the conditions in clinical practice. Nevertheless, this type of survey has its limitations.


The 1987 James A. F. Stevenson memorial lecture. The development of fetal behavioural patterns.
The development of fetal behaviour is reviewed. Fetal cutaneous and muscle sensory receptors are developed by the time movements are first seen. Human infants certainly respond to painful stimuli at 28 weeks. There is no clear evidence that prenatal "stress," e.g., maternal exposure to random noise and bright lights, impairs fetal development in the rat, on the contrary. Fetal diurnal rhythms appear in man and sheep before the development of sleep states; they are dependent on maternal corticosteroids, but the fetal mechanism is uncertain. With the development of sleep states (in late gestation in man and sheep, postnatally in the rat), the complex central control of behaviour is gradually established, but wakefulness is still of low incidence. The location of the sleep cycle generator is uncertain; the results of experimental lesions of the brainstem in fetal lambs appear incompatible with studies in adult rats and cats.


Fluctuations in accommodation: a review.
When a young observer attempts to accommodate steadily on a fixed stimulus, the nominally steady-state response shows small instabilities or fluctuations (sometimes termed microfluctuations or oscillations). These fluctuations typically have an amplitude of a few tenths of a dioptre and a frequency spectrum extending up to a few Hertz. The properties of these fluctuations are described for various viewing conditions: pupil diameter, target vergence, target form, target contrast, and target luminance all influence the frequency spectra of the oscillations, as may anomalies of vision such as amblyopia. The possible roles that the fluctuations might play in the function of the accommodative system are discussed. It is suggested that the higher frequency components around 2 Hz may arise from the mechanical and elastic characteristics of the lens, zonule and ciliary body. Components at lower frequencies (less than 0.5 Hz) may be of more significance in the function of the accommodative control system.


Lactobacillus plantarum RS-09 Induces M1-Type Macrophage Immunity Against Salmonella Typhimurium Challenge via the TLR2/NF-κB Signalling Pathway.
Lactobacillus plantarum can interact with macrophages against bacterial enteropathy due to its potential ability to modulate macrophage polarization. However, this mechanism is not completely understood. TLR2 can recognize microbial components and trigger macrophage cytokine responses to different gram-positive strains. The aim of this study was to investigate whether probiotic Lactobacillus plantarum RS-09 can induce macrophage polarization against Salmonella Typhimurium infection via TLR2 signalling. BALB/c mice were preadministered RS-09 continuously for 7 days and then infected with Salmonella Typhimurium ATCC14028. Mouse RAW264.7 mononuclear macrophages were stimulated with RS-09 and coincubated with ATCC14028 or PBS controls. The results of the in vivo study indicated that RS-09 could relieve S. Typhimurium-induced splenomegaly, body weight loss and death rate. RS-09 also limited the colonization and translocation of S. Typhimurium in the gastrointestinal tract and thereby protected against infection. We also observed that RS-09 upregulated the production of M1 macrophage characteristics (e.g., CD11c and IL-6) against S. Typhimurium. Furthermore, RS-09 induced the expression of TLR2 in macrophages. In an in vitro study, treatment of RAW264.7 cells with RS-09 either concurrently with or before S. Typhimurium challenge enhanced the secretion of Reactive oxygen species and Nitric oxide. This effect was related to TLR2 and NF-κB activation. Based on these findings, Lactobacillus plantarum RS-09 was shown to modulate M1 macrophage polarization and induce TLR2-linked NF-κB signalling activity in the innate immune response to S. Typhimurium infection.


Cloning and functional characterisation of the interleukin-1 beta 1 promoter of rainbow trout (Oncorhynchus mykiss).
The upstream flanking region of the rainbow trout (Oncorhynchus mykiss) IL-1 beta 1 gene has been cloned and characterised functionally using luciferase-based reporter gene constructs, and the transcription start site (TSS) confirmed by RLM-RACE. A TATA box was present 27 bp upstream of the TSS, with an NF-kB site 19 bp upstream of the TATA box. Within 1217 bp of upstream sequence, 3 sites for NF-kB, 10 sites for NF-IL6, 15 sites for AP1, 6 sites for AP4, 2 sites for CHOP/CEBP alpha and 1 site for SP1 and PU.1 were identified. Seven potential sites for the transcription repressor Gfi-1 were also identified. Analysis of eight IL-1 beta 1s promoter luciferase constructs transfected into a trout fibroblast (RTG-2) cell line known to constitutively express IL-1 beta revealed that in the absence of intron 1, very low luciferase activity was detectable. All of the constructs containing intron 1 gave clear luciferase activity, with the highest luciferase activity detected with construct P2-4 containing 617 bp of upstream sequence. As little as 82 bp of upstream sequence gave relatively strong luciferase activity, a region containing both a PU.1 and NF-kB site. That NF-kB is a transcription factor required for expression of the trout IL-1 beta 1 gene was confirmed using inhibitor studies with lipopolysaccharide (LPS)-stimulated macrophages. Both trout recombinant IL-1 beta and LPS were able to increase luciferase activity in the reporter constructs, especially in those containing the most upstream sequence with the lowest constitutive expression. The possibility that an upstream repressor is functioning to inhibit constitutive expression of IL-1 beta in this species is discussed.


Hybrid organic-inorganic anion-exchange pore-filled membranes for the recovery of nitric acid from highly acidic aqueous waste streams.
Recycling of acid from aqueous waste streams is highly important not only from the environmental point of view but also for developing the sustainable technology. One of the effective ways to recover acid from aqueous waste streams is the anion-exchange membrane based diffusion-dialysis. The work presents the synthesis and characterization of anion-exchange pore-filled membranes for the objective of recovery of high concentration of acid by diffusion dialysis. The membranes were prepared by anchoring the guest organic-inorganic anionic gel in the pores of the host poly(propylene) membrane by in situ UV-initiator induced polymerization of the appropriate monomers along with cross-linker. The removal of nitric acid in the presence of different representative monovalent, divalent and trivalent nitrates and the leakage of these ions through anion exchange membrane have been studied by DD technique for optimizing the chemical composition of the membrane. The nitric acid permeation rate of the membrane with the optimized composition has been found to be considerably faster than the commercial Selemion membrane without sacrificing salt leakage. The performance of the optimized pore-filled anion exchange membranes has been found to be independent of the acid concentration, nature of the anion and substrate and has been observed to be solely dependent on the guest inorganic-organic hybrid anionic gel component. The membranes have been found to be stable and reusable for the acid recovery. Removal of nitric acid as high as 90% from the simulated high level nuclear waste with the optimized grafted pore-filled membrane has been achieved with negligible salt transport.


Imaging, Pathology, and Immune Correlates in the Woodchuck Hepatic Tumor Model.
Woodchucks chronically infected with woodchuck hepatitis virus (WHV), which resembles human hepatitis B virus, develop spontaneous hepatic tumors and may be an important biological and immunological model for human HCC. Nonetheless, this model requires further validation to fully realize its translational potential. Woodchucks infected at birth with WHV that had developed HCC (n=12) were studied. Computed tomography, ultrasound, and magnetic resonance imaging were performed under anesthesia. LI-RADS scoring and correlative histologic analysis of sectioned tissues were performed. For immune characterization of tumors, CD3 (T cells), CD4 (T helpers), NCAM (Natural killers), FOXP3 (T-regulatory), PDL-1 (inhibitory checkpoint protein), and the human hepatocellular carcinoma (HCC) biomarker alpha-fetoprotein (AFP) immunohistochemical stains were performed. Forty tumors were identified on imaging of which 29 were confirmed to be HCC with 26 categorized as LR-4 or 5. The remainder of the tumors had benign histology including basophilic foci, adenoma, and lipidosis as well as pre-malignant dysplastic foci. LR-4 and LR-5 lesions showed high sensitivity (90%) and specificity (100%) for malignant and pre-malignant tumors. Natural killers count was found to be 2-5 times lower in tumors relative to normal parenchyma while other immune cells were located in the periphery of tumors. Tumors expressed AFP and did not express PD-L1. Woodchucks chronically infected with WHV developed diverse hepatic tumor types with diagnostic imaging, pathology, and immune patterns comparable to that in humans. This unique animal model may provide a valuable tool for translation and validation of novel image-guided and immune-therapeutic investigations.


Stability-indicating methods for the determination of famciclovir in the presence of its alkaline-induced degradation product.
Five sensitive, selective and precise stability-indicating methods are presented for the determination of famciclovir (FCV) in the presence of its alkaline-induced degradation product. Method A utilizes the first derivative spectrophotometry at 321 nm. Method B depends on using the first derivative of the ratio spectrophotometry (DD(1)) by measurement of the amplitude at 256 nm. Method C is based on the reaction of FCV with hydroxylamine to form hydroxamic acid, causing the hydroxamic acid to react with triferric ion to form ferric hydroxamate that is measured at 503 nm. Method D is based on the separation of FCV from its degradation product followed by densitometric measurement of the bands at 304 nm. The separation was carried out on silica gel 60 F(254), using chloroform: methanol (70:30, v/v) as a mobile phase. Method E is based on a high performance liquid chromatographic (HPLC) separation of FCV from its degradation product using an ODS column with a mobile phase consisting of methanol-50 mM dipotassium hydrogen phosphate (25:75, v/v, pH 3.0)with UV detection at 304 nm. Regression analysis showed good correlation in the concentration ranges 16-72 microg/ml, 40-240 microg/ml, 40-240 microg/ml, 0.75-5.25 microg/band and 20-240 microg/ml with percentage recoveries of 99.65 +/- 0.85, 100.27 +/- 0.91, 99.72 +/- 0.84, 100.65 +/- 1.52 and 99.88 +/- 0.50 for methods A, B, C, D and E, respectively. These methods are suitable as stability-indicating methods for the determination of FCV in the presence of its degradation product either in bulk powder or in pharmaceutical formulation. Statistical analysis of the results has been carried out revealing high accuracy and good precision.


Demetallization of Sewage Sludge Using Low-Cost Ionic Liquids.
Sludge produced from wastewater treatment has little to no value and is typically treated through volume reduction techniques, such as dewatering, thickening, or digestion. However, these methods inherently increase heavy metal concentrations, which makes the sludge unsuitable for land spreading and difficult to dispose of, owing to strict legal requirements/regulations concerning these metals. We addressed this problem, for the first time, by using recyclable low-cost protic ionic liquids to complex these toxic metals through a chemical fractionation process. Sewage sludge samples collected from wastewater plants in the UK were heated with methylimidazolium chloride ([Hmim]Cl, triethylammonium hydrogen sulfate ([TEA][HSO4]) and dimethylbutylammonium hydrogen sulfate ([DMBA][HSO4]) under various operating temperatures, times and solids loadings to separate the sludge from its metal contaminants. Analysis of the residual solid product and metal-rich ionic liquid liquor using inductively coupled plasma-emission spectrometry showed that [Hmim]Cl extracted >90% of CdII, NiII, ZnII, and PbII without altering the phosphorus content, while other toxic metals such as CrIII, CrVI and AsIII were more readily removed (>80%) with [TEA][HSO4]. We test the recyclability of [Hmim]Cl, showing insignificant efficiency losses over 6 cycles and discuss the possibilities of using electrochemical deposition to prevent the buildup of metal in the IL. This approach opens up new avenues for sewage sludge valorization, including potential applications in emulsion fuels or fertilizer development, accessed by techno-economic analysis.


Cytomegalovirus retinitis in rheumatic disease: a case report.
We describe a 67-year-old white man with Wegener's granulomatosis who was treated with cyclophosphamide, then with azathioprine, and who subsequently developed cytomegalovirus retinitis. The patient initially improved on a course of adenine arabinosine, but his retinitis has progressed, despite discontinuation of immunosuppressive therapy. We show that cytomegalovirus retinitis does occur in patients with rheumatic diseases who receive immunosuppressive therapy.


Structural mass spectrometry analysis of lipid changes in a Drosophila epilepsy model brain.
Phosphatidylethanolamine (PtdEtn) is one of the most abundant phospholipids in many animal cell types. The Drosophila easily shocked (eas(2)) mutant, used as an epilepsy model, is null for the PtdEtn biosynthetic enzyme, ethanolamine kinase. This mutant displays bang sensitive paralysis, and was previously shown to have decreased levels of PtdEtn. We have developed a highly selective and sensitive measurement strategy using ion mobility-mass spectrometry for the relative quantitation of intact phospholipid species directly from isolated brain tissue of eas mutants. Over 1200 distinct lipid signals are observed and within this population 38, including PtdEtn, phosphatidylinositol (PtdIns) and phosphatidylcholine (PtdCho) species are identified to have changed significantly (p < 0.03) between mutant and control tissue. This method has revealed for the first time the structural complexity and biosynthetic interconnectedness of specific PtdEtn and PtdIns lipid species within tissue, and provides great molecular detail compared to traditionally used detection techniques.


5-azacytidine enhances efficacy of multiple chemotherapy drugs in AML and lung cancer with modulation of CpG methylation.
The DNA methyltransferase (DNMT) inhibitory drugs such as 5-azacytidine induce DNA hypomethylation by inhibiting DNA methyltransferases. While clinically effective, DNMT inhibitors are not curative. A combination with cytotoxic drugs might be beneficial, but this is largely unexplored. In the present study, we analyzed potential synergisms between cytotoxic drugs and 5-azacytidine in acute myeloid leukemia (AML) and non-small cell lung cancer (NSCLC) cells. Lung cancer and leukemia cell lines were exposed to low doses of 5-azacytidine with varying doses of cytarabine or etoposide for AML cells (U937 and HL60) as well as cisplatin or gemcitabine for NSCLC cells (A549 and HTB56) for 48 h. Drug interaction and potential synergism was analyzed according to the Chou-Talalay algorithm. Further analyses were based on soft agar colony formation assays, active caspase-3 staining and BrdU incorporation flow cytometry. To identify effects on DNA methylation patterns, we performed genome wide DNA methylation analysis using 450K bead arrays. Azacytidine at low doses was synergistic with cytotoxic drugs in NSCLC and in AML cell lines. Simultaneous exposure to 5-azacytidine with cytotoxic drugs showed strong synergistic activity. In colony formation assays these synergisms were repeatedly verified for 5-azacytidine (25 nM) with low doses of anticancer agents. 5-azacytidine neither affected the cell cycle nor increased apoptosis. 450K methylation bead arrays revealed 1,046 CpG sites in AML and 1,778 CpG sites in NSCLC cells with significant DNA hypomethylation (24-h exposure) to 5-azacytidine combined with the cytotoxic drugs. These CpG-sites were observed in the candidate tumor-suppressor genes MGMT and THRB. Additional incubation time after 24-h treatment led to a 4.1-fold increase of significant hypomethylated CpG-sites in NSCLC cells. These results suggest that the addition of DNA demethylating agents to cytotoxic anticancer drugs exhibits synergistic activity in AML and NSCLC. Dysregulation of an equilibrium of DNA methylation in cancer cells might increase the susceptibility for cytotoxic drugs.


Efficacy of Plectranthus glandulosus (Lamiaceae) and Callistemon rigidus (Myrtaceae) Leaf Extract Fractions to Callosobruchus maculatus (Coleoptera: Bruchidae).
As part of on-going efforts to use eco-friendly alternatives to chemical pesticides, methanol crude extracts of Plectranthus glandulosus and Callistemon rigidus leaves were sequentially fractionated in hexane, chloroform, ethyl acetate, and methanol to establish the most active fraction(s) against Callosobruchus maculatus in cowpea. Cowpea seeds (25 g) were treated with 0.5, 1, 2, and 4 g/kg of extract to evaluate the contact toxicity and F1 progeny production of the beetles in the laboratory. Mortality was recorded 1, 3, and 7 d postexposure. P. glandulosus hexane fraction was more toxic than the other fractions recording 100% mortality at 4 g/kg, within 7 d with LC50 of 0.39 g/kg. Hexane fraction of C. rigidus showed superior toxicity, causing 100% mortality at 4 g/kg within only 1 d of exposure with LC50 of 1.02 g/kg. All the fractions greatly reduced progeny emergence, with C. rigidus hexane fraction being the best progeny inhibitor. Fractions of P. glandulosus and C. rigidus leaves had sufficient efficacy to be a component of storage pest management package for C. maculatus.


Ring expansion of cyclic β-amino alcohols induced by diethylaminosulfur trifluoride: synthesis of cyclic amines with a tertiary fluorine at C3.
As the replacement of a hydrogen atom by a fluorine atom in a compound can have an important impact on its biological properties, the development of methods allowing the introduction of a fluorine atom is of great importance. The scope and limitations of the ring expansion of cyclic 2-hydroxymethyl amines induced by diethylaminosulfur trifluoride (DAST) to produce cyclic β-fluoro amines was studied as well as the enantioselectivity of the process.


Stimulus-Induced Motor Afterdischarges in CASPR2 (Contactin-Associated Protein-Like 2)-Positive Peripheral Nerve Hyperexcitability Syndrome.
Peripheral nerve hyperexcitability is an uncommon but treatable condition in neurology. Voltage-gated potassium channelopathies, especially contactin-associated protein-like 2 (CASPR2) antibody, are commonly implicated. We present the case of a 16-year-old boy with tremulousness of both feet and twitching of muscles all over the body for three months. Examination revealed irregular, arrhythmic, small-amplitude twitching movements of the toes along with fasciculations in both thighs. Nerve conduction studies were within normal limits. F-wave studies showed a prolonged polyphasic large-amplitude discharge following the compound muscle action potential and obscuring the F waves. Electromyography showed extensive myokymic discharges. The serum autoimmune antibody profile showed strong positivity for CASPR2. He started lacosamide as a symptomatic treatment. In view of the good symptomatic response, further immunomodulation was deferred and he remains on follow-up. We present this case to highlight the role of motor afterdischarges as a diagnostic clue to peripheral nerve hyperexcitability and to review the literature on this interesting finding.


Mavacamten, an Alternative to Septal Reduction Therapy for Patients with Hypertrophic Cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a common heridetary cardiac disorder characterized by a wide range of symptoms. The pharmacological treatment of HCM is currently limited to beta blockers, non-dihydropyridine calcium channel blockers and disopyramide. Mavacamten is a novel cardiac myosin inhibitor, which was recently added to the limited pharmacological list of treatment options for HCM. This editorial elaborates on current evidence evaluating the use of mavacamten in patients with symptomatic obstructive HCM, comments on its current use and its expanded potential applications in the future.


[Epithelial cadherins and associated molecules in invasive lobular breast cancer].
to estimate the expression of cell adhesion molecules E- and P-cadherin, as well as that of cadherin-catenin complexes in invasive lobular breast cancer (BC) cells. 250 cases of postoperative material from patients diagnosed with invasive lobular BC were studied. The expressions of cell adhesion molecules E-cadherin, P-cadherin, β-catenin, p120 catenin, and vimentin were determined by immunohistochemical assay in all cases. The examined cases were divided into molecular biological subtypes, based on the evaluation of estrogen receptors (ER), progesterone receptors (PR), HER-2/neu, and Ki-67 proliferative index. The membrane expression of E-cadherin on the tumor cells was found to be preserved in 93%; the cytoplasmic expression of β-catenin and p120-catenin appeared in 60 and 72% of cases, respectively. The expression of P-cadherin was detected in 82% of cases. The coexpression of E- and P-cadherin was noted in 90% of all the examined cases. There was a correlation between the expression of E- and P-cadherins (V=0.34; p<0.05). The BC cells showed the coexpression of E- and P-cadherins, as well as release of the molecules β- and p120-catenins into the cytoplasm of tumor cells, which leads to the activation of intracellular mechanisms for changing the structure of the cytoskeleton and the level of proliferation. The above-mentioned mechanisms are accompanied by the activation of epithelial-mesenchymal transition. The intracellular mechanisms resulted in progressive cancer and its metastasis.


Promoting an ethic of engagement in pediatric palliative care research.
This paper defends the ethical and empirical significance of direct engagement with terminally ill children and adolescents in PPC research on health-related quality of life. Clinical trials and other forms of health research have resulted in tremendous progress for improving clinical outcomes among children and adolescents diagnosed with a life-threatening illness. Less attention has been paid, however, to engaging this patient population directly in studies aimed at optimizing health-related quality of life in PPC. Though not restricted to care at the end of life, PPC--and by extension PPC research--is in part dependent on recognizing the social complexities of death and dying and where health-related quality of life is a fundamental element. To explore these complexities in depth requires partnership with terminally ill children and adolescents, and acknowledgement of their active social and moral agency in research. Principles of pediatric research ethics, theoretical tenets of the "new sociology of the child(hood)," and human rights codified in the United Nations Convention on the Rights of the Child (UNCRC) underpin the position that a more engagement-centered approach is needed in PPC research. The ethics, sociologies and human rights of engagement will each be discussed as they relate to research with terminally ill children and adolescents in PPC. Qualitative method(ologies) presented in this paper, such as deliberative stakeholder consultations and phenomenology of practice can serve as meaningful vehicles for achieving i) participation among terminally ill children and adolescents; ii) evidence-bases for PPC best practices; and iii) fulfillment of research ethics principles. PPC research based on direct engagement with PPC patients better reflects their unique expertise and social epistemologies of terminal illness. Such an approach to research would strengthen both the ethical and methodological soundness of HRQoL inquiry in PPC.


The economic burden of spinal muscular atrophy.
To evaluate the economic burden of spinal muscular atrophy (SMA). This study used Department of Defense Military Healthcare System (MHS) data from 2003-2012. Healthcare costs were determined for patients with at least one inpatient or three outpatient claims with a diagnosis of SMA before 18 years of age and who had ≥ 6 months of data after first SMA diagnosis or expired within 6 months of initial diagnosis. A comparator cohort was selected using a 3:1 match based on age and gender. A total of 239 individuals with SMA diagnosis met the inclusion criteria along with 717 matched comparator patients. More patients with SMA had hospitalizations (69.5%) compared to the comparator cohort (17.2%, p < 0.001). Median total expenditures across all years of data for patients with SMA were $83 652 (25-75th percentile = $29 620-228 754) vs the comparator group of $4329 (25-75(th) percentile = $1229-10 062 (p < 0.001)) over an average (SD) of 6.9 ± 3.6 years. The annualized mean costs of total healthcare expenditures were significantly higher for the SMA cases than the comparison cohort, $47 862 ± 88 607 compared to $1861 ± 6374, respectively (p < 0.001). The sub-group of patients with early diagnosis (n = 45) had 4.3 ± 2.9 years of observation with a median cost of $167 921 ($53 349-678 412). Mean age (SD) at first observed SMA diagnosis was 7.5 ± 6.4 years. Mean (SD) duration of follow-up after initial SMA diagnosis was 4.8 ± 3.3 years, with a median post-diagnosis cost of $60 213 ($18 229-192 559). The major costs for all patients were outpatient visits [median = $53 152 ($23 902-136 150)], followed by inpatient costs [median = $11 258 ($0-51 987)] and total prescription costs [median = $3167 ($943-13 283)]. The analysis is limited to the data available and may under-estimate the total cost of SMA. Individuals with SMA have a high degree of morbidity, particularly those diagnosed during infancy. SMA patients have significant medical expenditures and high utilization of healthcare services.


Evaluation of systemic exposure of nanoparticle suspensions subcutaneously administered to mice regarding stabilization, volume, location, concentration and size.
Different routes of administration are likely to result in very different outcomes due to different availability or plasma profile. The objective of the present study was to evaluate the pharmacokinetic profile after different subcutaneous (s.c.) administration of nanoparticle suspensions of a lipophilic compound to mice. Pharmacokinetics of the selected test compound and the effect of drug concentration, particle size, location of administration, volume given and particle stabilizers were studied. Adding PEGylated lipids or pluronic F-127 to the negatively charged surface of the nanoparticles increased the stability of the particles and the bioavailability. The in vivo studies demonstrated linear absorption kinetics for the selected model compound up to at least 500 µmol/kg. Absorption from upper-neck resulted in different systemic exposure compared to administration in the hip. The former was preferred if a prolonged Cmax was desired while the latter ensured a flat profile for approximately 24 hours. Administering the double volume (but the same dose) had no effect on the pharmacokinetics, whereas smaller particle size significantly increased the exposure.


Empowering nurses for work engagement and health in hospital settings.
Employee empowerment has become an increasingly important factor in determining employee health and wellbeing in restructured healthcare settings. The authors tested a theoretical model which specified the relationships among structural empowerment, 6 areas of worklife that promote employee engagement, and staff nurses' physical and mental health. A predictive, non-experimental design was used to test the model in a random sample of staff nurses. The authors discuss their findings and the implication for nurse administrators.


Assessment of abdominal muscle function in individuals with motor-complete spinal cord injury above T6 in response to transcranial magnetic stimulation.
To use transcranial magnetic stimulation and electromyography to assess the potential for preserved function in the abdominal muscles in individuals classified with motor-complete spinal cord injury above T6. Five individuals with spinal cord injury (C5-T3) and 5 able-bodied individuals. Transcranial magnetic stimulation was delivered over the abdominal region of primary motor cortex during resting and sub-maximal (or attempted) contractions. Surface electromyography was used to record motor-evoked potentials as well as maximal voluntary (or attempted) contractions in the abdominal muscles and the diaphragm. Responses to transcranial magnetic stimulation in the abdominal muscles occurred in all spinal cord injury subjects. Latencies of muscle response onsets were similar in both groups; however, peak-to-peak amplitudes were smaller in the spinal cord injury group. During maximal voluntary (or attempted) contractions all spinal cord injury subjects were able to elicit electromyography activity above resting levels in more than one abdominal muscle across tasks. Individuals with motor-complete spinal cord injury above T6 were able to activate abdominal muscles in response to transcranial magnetic stimulation and during maximal voluntary (or attempted) contractions. The activation was induced directly through corticospinal pathways, and not indirectly by stretch reflex activations of the diaphragm. Transcranial magnetic stimulation and electromyography measurements provide a useful method to assess motor preservation of abdominal muscles in persons with spinal cord injury.


Rm values of phenols. Their relationship with log P values and activity.
The experimental Rm values for a series of phenols were obtained by a reversed-phase TLC system. The extrapolation from a range of linear relationship between experimental Rm values and acetone concentration provided a set of extrapolated Rm values. This were used for studying the relationship between structure and activity in vitro and in vivo. The possibility to obtain by means of the extrapolation technique the Rm values in a standard system for serveral series of chemotherapeutic agents is pointed out.


Distinguishing the Chinese materia medica Tiepishihu from similar Dendrobium species of the same genus using histological and microscopic method.
The Chinese Materia Medica, Tiepishihu, used as a tonic for over one thousand years, is a well-known precious medicine in China. According to the Chinese Pharmacopoeia, its source is the species Dendrobium officinale Kimura et Migo, which is distinguished from other species in Dendrobium genus. However, these species from the same genus are similar with Tiepishihu and caused confusion in the market. To find a quick and simple method to distinguish Tiepishihu from other similar species, histologic and microscopic methods were combined together to investigate the transverse section of stem of Tiepishihu and other similar species. Phloroglucinol test solution with hydrochloric acid was used to reveal the lignified tissue by staining the transverse section of Tiepishihu and similar species. Results revealed the unique identification characteristics to distinguish Tiepishihu from similar species, which were difficult to distinguish by other methods. The identification characteristics of Tiepishihu include the cells of vascular bundle sheath were stained red, parenchyma cells were not stained red. What's more, other species can be distinguished from each other with microscopic and histological characteristics. These characteristics proved stable and can be easily observed by normal light microscopic examination. This method is rapid, accurate, stable, and inexpensive.


Childhood maltreatment and the development of relational and physical aggression: the importance of a gender-informed approach.
This investigation examined the associations between maltreatment and aggression using a gender-informed approach. Peer ratings, peer nominations, and counselor reports of aggression were collected on 211 maltreated and 199 nonmaltreated inner-city youth (M age = 9.9 years) during a summer day camp. Maltreatment was associated with aggressive conduct; however, these effects were qualified by gender, maltreatment subtype, and the form of aggression under investigation. Findings revealed that maltreatment was associated with physical aggression for boys and relational aggression for girls. Physical abuse was associated with physically aggressive behaviors, but sexual abuse predicted relational aggression for girls only. Findings suggest that investigating the interaction between familial risk and gender is important in understanding aggressive behaviors of boys and girls.


A Prospective Evaluation of Circulating Tumor Cells and Cell-Free DNA in EGFR-Mutant Non-Small Cell Lung Cancer Patients Treated with Erlotinib on a Phase II Trial.
Genotype-directed therapy is the standard of care for advanced non-small cell lung cancer (NSCLC), but obtaining tumor tissue for genotyping remains a challenge. Circulating tumor cell (CTC) or cell-free DNA (cfDNA) analysis may allow for noninvasive evaluation. This prospective trial evaluated CTCs and cfDNA in EGFR-mutant NSCLC patients treated with erlotinib until progression. EGFR-mutant NSCLC patients were enrolled in a phase II trial of erlotinib. Blood was collected at baseline, every 2 months on study, and at disease progression. Plasma genotyping was performed by droplet digital PCR for EGFR19del, L858R, and T790M. CTCs were isolated by CellSave, enumerated, and analyzed by immunofluorescence for CD45 and pan-cytokeratin and EGFR and MET FISH were also performed. Rebiopsy was performed at disease progression. Sixty patients were enrolled; 44 patients discontinued therapy for disease progression. Rebiopsy occurred in 35 of 44 patients (80%), with paired CTC/cfDNA analysis in 41 of 44 samples at baseline and 36 of 44 samples at progression. T790M was identified in 23 of 35 (66%) tissue biopsies and 9 of 39 (23%) cfDNA samples. CTC analysis at progression identified MET amplification in 3 samples in which tissue analysis could not be performed. cfDNA analysis identified T790M in 2 samples in which rebiopsy was not possible. At diagnosis, high levels of cfDNA but not high levels of CTCs correlated with progression-free survival. cfDNA and CTCs are complementary, noninvasive assays for evaluation of acquired resistance to first-line EGFR TKIs and may expand the number of patients in whom actionable genetic information can be obtained at acquired resistance. Serial cfDNA monitoring may offer greater clinical utility than serial monitoring of CTCs. Clin Cancer Res; 22(24); 6010-20. ©2016 AACR.


Mental health literacy about schizophrenia and depression: a survey among Chinese caregivers of patients with mental disorder.
To investigate the knowledge of schizophrenia and depression among caregivers of patients with mental disorder in China. A convenience sample of 402 caregivers at the Department of Psychiatry of a general hospital in China was investigated (response rate 95.7%), using vignettes based investigation methodology. The number of caregivers using the term "depression" to describe the depression vignette was 43.6%, which was significantly higher than the number of caregivers using the term "schizophrenia" to describe the schizophrenia one (28.5%). A high percentage of caregivers believed that "psychiatrist", "psychologist" and "close family members" would be helpful, and the top three most helpful interventions were "becoming more physically active", "getting out and learning more" and "receiving psychotherapy". The number of caregivers endorsed "antipsychotics" and "antidepressants" as helpful for the schizophrenia and the depression vignettes were 82.0 and 80.7%, respectively. Regarding the causes of mental illness, items related to psychosocial factors, including "daily problems" and "work or financial problems", and "weakness of character" were highly rated, with half considered genetic or chemical imbalance causes. Caregivers expressed a high knowledge about treatments and interventions of mental disorders. But there are still some areas, particularly regarding the recognition and causes of mental disorders, that are in need of improvement. This is particularly the case for schizophrenia.


Prevalence of carriage of Shiga toxin-producing Escherichia coli serotypes O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28 among slaughtered adult cattle in France.
The main pathogenic enterohemorrhagic Escherichia coli (EHEC) strains are defined as Shiga toxin (Stx)-producing E. coli (STEC) belonging to one of the following serotypes: O157:H7, O26:H11, O103:H2, O111:H8, and O145:H28. Each of these five serotypes is known to be associated with a specific subtype of the intimin-encoding gene (eae). The objective of this study was to evaluate the prevalence of bovine carriers of these "top five" STEC in the four adult cattle categories slaughtered in France. Fecal samples were collected from 1,318 cattle, including 291 young dairy bulls, 296 young beef bulls, 337 dairy cows, and 394 beef cows. A total of 96 E. coli isolates, including 33 top five STEC and 63 atypical enteropathogenic E. coli (aEPEC) isolates, with the same genetic characteristics as the top five STEC strains except that they lacked an stx gene, were recovered from these samples.O157:H7 was the most frequently isolated STEC serotype. The prevalence of top five STEC (all serotypes included) was 4.5% in young dairy bulls, 2.4% in young beef bulls, 1.8% in dairy cows, and 1.0% in beef cows. It was significantly higher in young dairy bulls (P<0.05) than in the other 3 categories. The basis for these differences between categories remains to be elucidated. Moreover,simultaneous carriage of STEC O26:H11 and STEC O103:H2 was detected in one young dairy bull. Lastly, the prevalence of bovine carriers of the top five STEC, evaluated through a weighted arithmetic mean of the prevalence by categories, was estimated to 1.8% in slaughtered adult cattle in France.


[Dependence of EGF receptor and STAT factor activation on redox of A431 cells].
Reactive oxygen species (ROS) were established to play an important role in cellular signaling as second messengers by integrating different pathways. Recently, we showed that EGF initiated a rapid tyrosine phosphorylation of both EGF-receptor and STAT factors with simultaneous increase in the intracellular ROS level. Now, we have investigated the effect of intracellular red-ox state on EGF- and H2O2-induced activation of EGF receptor, STAT1 and STAT3. We demonstrated that the pretreatment of A431 cells with antioxidant N-acetyl-L-cysteine (NAC) partly reduced the level of EGF-induced phosphorylation of proteins under investigation. Besides, H2O2-induced activation of EGF receptor, and STAT factors was fully prevented by NAC pretreatment. The inhibition of ROS generation by DPI declined EGF-dependent activation of EGF receptor and STAT factors to basal level. Our results demonstrate the essential role of cellular red-ox status in the modulation of EGF-mediated activation of receptor and STAT factors. We have postulated that EGF-induced ROS generation is a very important initial event promoting physiological activation of EGF receptor and subsequent STAT factor activation.


Clinical trial design issues: at least 10 things you should look for in clinical trials.
Randomized controlled trials remain the gold standard study design and yield the highest level of scientific credence. However, recognition of the limitations of the randomized controlled trial is important. This review highlights 10 potentially problematic areas one should carefully assess when performing or reading an article reporting the results of a randomized controlled trial, problematic areas that can affect the outcome of the trial and therefore mislead the reader. These areas include ethical issues, eligibility criteria, masking (blinding), randomization, analytic methods, the selection of subjects for the interventional and comparison groups, selection of end points, and the interpretation of the results. Each of these is discussed, and examples of published articles are used to highlight the main points.


External validation of the international mission for prognosis and analysis of clinical trials model and the role of markers of coagulation.
Markers of coagulation have shown to have important value in predicting traumatic brain injury outcome. To externally validate and investigate the role of markers of coagulation for outcome prediction by using the International Mission for Prognosis and Analysis of Clinical Trials (IMPACT) model while adjusting for overall injury severity. A retrospective chart analysis of traumatic brain injury patients admitted to Helsinki University Central Hospital between 2009 and 2010 was performed. Outcome was estimated by using the criteria of the IMPACT model. Admission international normalized ratio (INR) and platelet count were used as markers of coagulation. Overall injury severity was categorized with the injury severity score (ISS). Variables were added to the calculated IMPACT risk, generating new models. Model performance was assessed by analyzing and comparing the area under the curve (AUC) of the models. For 342 included patients, 6-month mortality was 32% and unfavorable neurological outcome was 36%. Patients with a poor outcome had lower platelets and higher INR and ISS than those with good outcome (P < .001). The IMPACT model had an AUC of 0.85 for predicting mortality and 0.81 for neurological outcome. Addition of INR but not ISS or platelets to the IMPACT predicted risk improved the predictive validity for mortality ([INCREMENT]AUC 0.02, P = .034) but not neurological outcome ([INCREMENT]AUC 0.00, P = .401). In multivariate analysis, INR remained significant for mortality but not for neurological outcome when adjusting for IMPACT risk and ISS (P = .012). The IMPACT model showed excellent performance, and INR was an independent predictor for mortality, independent of overall injury severity.


Engagement of CD4 and CD8 accessory molecules is required for T cell maturation.
In order to address the role of CD4 and CD8 Ag in the process of positive selection in the thymus, antibodies against these molecules, which do not result in the elimination of mature lymph node T cells, were injected in vivo. The results indicate that even long-term injection of nondepleting anti-CD4 and anti-CD8 antibodies does not cause the loss of CD4 or CD8 positive lymph node cells, but it completely blocks the development of the corresponding subpopulation of mature thymocytes. Thus, it appears that the interaction of the CD4 and CD8 accessory molecules on developing thymocytes with a ligand in the thymic environment (probably MHC Ag) is necessary for the positive selection of thymocytes into the appropriate T cell lineage.


Chronic intracerebroventricular infusion of insulin failed to alter brain insulin-binding sites, food intake, and body weight.
The present study was performed to explore the role of exogenous insulin in CSF in the control of energy balance in the rat. For this purpose, adult male Sprague-Dawley rats carrying an indwelling cannula in the right lateral cerebral ventricle were infused for a maximum of 10 days with insulin (Actrapid) at various rates (starting at 0, 45, 85, 170, and 600 ng/day) or anti-insulin antibody (IgG fraction; diluted 1:10 wt/vol) with an osmotic minipump. All those treatments did not modify the growing rates; neither total daily food intake nor the circadian rhythm of food intake was further modified. The chronic insulin infusion starting at 600 ng/day resulted in a chronic significant increase in CSF insulin levels without changing the plasma insulin level. It failed to alter specific insulin binding sites to Triton X-100 solubilized microsomal membranes from various brain areas (cerebral cortex, olfactory bulbs, and lateral and medial hypothalami) at the end of the 5- or 10-day period of insulin infusion. Purification of insulin receptors on a wheat germ agglutinin did not reveal any further effect of insulin. From these results, it seems unlikely that the input to the brain insulin-effector systems could arise from CSF insulin.


Personalized medicine and tobacco-related health disparities: is there a role for genetics?
Genetic testing has been proposed as a means to increase smoking cessation rates and thus reduce smoking prevalence. To understand how that might be practically possible, with appreciation of the current social context of tobacco use and dependence, we performed a contextual analysis of smoking-related genetics and smoking cessation. To provide added value, genetics would need to inform and improve existing interventions for smokers (including behavioral and pharmacological treatments). Pharmacogenetics offers the most promising potential, because it may improve the efficacy of medication-based smoking cessations strategies. All proven interventions for treating tobacco dependence, however, including simple cost-effective measures, such as quit lines and physician counseling, are underutilized. As tobacco use occurs disproportionately among disadvantaged populations, efforts to improve smokers' access to health care, and to the tools that are known to help them quit, represent the most promising approaches for reducing smoking prevalence within these groups. Similar considerations apply to other chronic diseases contributing to population-level health disparities. We conclude that although genetics offers increasing opportunities to tailor drug treatment, and may in some cases provide useful risk prediction, other methods of personalizing care are likely to yield greater benefit to populations experiencing health disparities related to tobacco use.


Hyperacute rejection of guinea-pig to rat cardiac xenografts. I. Morphology.
The morphological changes occurring in xenograft hyperacute rejection, using guinea-pig to rat cardiac transplants, have been studied. Ultrastructural evidence of platelet aggregation in myocardial capillaries and veins first occurred two minutes after revascularisation. Small amounts of fibrin, associated with platelet degranulation occurred at three minutes. From four minutes, platelet aggregates increased in number and endothelial damage first appeared in those vessels containing platelet aggregates. Rejection was complete by 12 minutes and platelet aggregates then filled many capillaries, veins, arterioles and arteries. Polymorphonuclear leucocyte infiltration was not seen.


[Evaluation of the effectiveness of ultratonotherapy in patients with microbial eczema].
Microbial eczema is a chronic dermatosis with a complex pathogenesis. The result of exposure to various endo- and exogenous trigger factors is the formation of skin dysbiosis in lesions with a predominance of opportunistic and pathogenic flora and, as a result, the development of sensitization. As the main therapy for dysbiotic disorders, it is customary to use antibacterial drugs. However, often the use of systemic antibacterial drugs is not justified and leads to an aggravation of the problem of antibiotic resistance. To evaluate the effectiveness of the use of alternating current on the skin microbiome in the complex therapy of patients with microbial eczema. 2 groups of 30 patients with chronic moderate course of microbial eczema in the acute stage were examined. In the comparison group, therapy was carried out in accordance with the Federal Clinical Guidelines (antihistamines, detoxification, systemic antibacterial drugs, external combined agents, including a corticosteroid and an antibiotic). Patients of the main group received similar therapy without systemic antibiotics. In addition, to influence the pathogenic microflora of the skin in the lesion in the main group, procedures with supratonal alternating current of low strength were also used. The control group consisted of 30 healthy volunteers. The results were evaluated according to the clinical picture, laboratory parameters of bacteriological examination. During therapy, in patients of the main group and in the comparison group, a positive dynamics of the skin pathological process was noted, and a decrease in microbial colonization of the skin in the lesions of microbial eczema was observed by 4 and 3 times, respectively (p<0.05). The proposed method of treatment is not inferior in terms of the effectiveness of traditional therapy, allows to achieve positive clinical dynamics in similar terms, as well as to avoid the widespread use of systemic antibacterial drugs in cases of moderate dermatosis.


Tunable Volatile-Organic-Compound Sensor by Using Au Nanoparticle Incorporation on MoS2.
Controlling the charge concentrations of two-dimensional (2D) materials is a critical requirement for realizing versatility and potential application of these materials in high-performance electronics and sensors. In order to exploit the novel chemical-sensing characteristics of 2D materials for sensitive and selective sensors, various functionalization methods are needed to ensure efficient doping of channels based on 2D materials. In the present study, the gas-sensing performance of MoS2 has been significantly enhanced by controlled Au nanoparticle functionalization. By using the difference in reduction potential between the Au precursor and MoS2 work functions, MoS2 prepared by chemical exfoliation process was decorated with nanoparticles with sizes of tens of nanometers. The n-doping effect of Au nanoparticles was observed, that is, these particles were found to have facilitated in electron charge transfer from Au to MoS2. The controlled n-doping effect enables the tuning of the sensing of hydrocarbon-based volatile organic compounds (VOCs) and oxygen-functionalized compounds by MoS2. A significant step has therefore been made with this study toward solving the limitations imposed by previous MoS2-based sensors, which mostly produce a single response to various VOC analytes. This controllable chemical doping process for tuning the VOC-sensing performance of MoS2 can eventually be used in early detection using multichannel sensing systems that have different responses and recognize patterns for target analytes.


Cultivated ginseng inhibits 2,4-dinitrochlorobenzene-induced atopic dermatitis-like skin lesions in NC/Nga mice and TNF-α/IFN-γ-induced TARC activation in HaCaT cells.
Ginseng contains many bioactive constituents, including various ginsenosides that are believed to have anti-allergic, anti-oxidant, and immunostimulatory activities; however, its effects on atopic dermatitis (AD) remain unclear. In the current study, we hypothesized that cultivated ginseng (CG) would inhibit 2,4-dinitrochlorobenzene (DNCB)-induced AD-like skin lesions in NC/Nga mice by regulating the T helper (Th)1/Th2 balance. Also, CG inhibits TNF-α/IFN-γ-induced thymus- and activation-regulated chemokine (TARC) expression through nuclear factor-kappa B (NF-κB)-dependent signaling in HaCaT cells. CG ameliorated DNCB-induced dermatitis severity, serum levels of IgE and TARC, and mRNA expression of TARC, TNF-α, IFN-γ, IL-4, IL-5, and IL-13 in mice. Histopathological examination showed reduced thickness of the epidermis/dermis and dermal infiltration of inflammatory cells in the ears. Furthermore, CG suppressed the TNF-α/IFN-γ-induced mRNA expression of TARC in HaCaT cells. CG inhibited TNF-α/IFN-γ-induced NF-κB activation. These results suggest that CG inhibited the development of the AD-like skin symptoms by modulating Th1 and Th2 responses in the skin lesions in mice and TARC expression by suppressing TNF-α/IFN-γ-induced NF-κB activation in keratinocytes, and so may be a useful tool in the therapy of AD-like skin symptoms.


The carbon monoxide diffusing capacity: clinical implications, coding, and documentation.
The test for the diffusing capacity of the lung for carbon monoxide (DLCO) has been available for nearly 100 years for research and clinical purposes. The single-breath method is used almost exclusively in the United States It has been available in clinical pulmonary function laboratories for > 50 years. DLCO has great value in evaluating patients with lung diseases. Guidelines to standardize DLCO have been published by the American Thoracic Society and European Respiratory Society to reduce the interlaboratory variability that has existed. One code, 94720, should be reported for the billing for DLCO. Another code, 94725, the membrane diffusing capacity, exists for the measurement of the membrane and blood components of the DLCO. Currently, no clinical indications exist for the use of the membrane diffusing capacity. The finding that the number of tests in the Medicare population coded with 94725 has increased by > 1,000% from 2000 to 2005 is quite surprising. This rate is 14-times higher than the rate of increase in the utilization of 94720 over the same period. The possible reasons for these increases are discussed, but the most likely explanation is the financial gain derived from coding 94725. It is proposed that coding and billing of 94725 be stopped until the clinical indications for membrane diffusing capacity have been established. Those who code and bill for 94725 must be prepared to justify the use of this code to Medicare and third-party payers.


Exploring thermodynamic and structural properties of carbon nanotube/thermoplastic polyurethane nanocomposites from atomistic molecular dynamics simulations.
Carbon nanotubes (CNTs) and thermoplastic polyurethane (TPU) nanocomposites have emerged as promising materials for various applications in the field of nanotechnology. An understanding of the thermodynamic and structural properties is of fundamental significance in designing and fabricating CNT/TPU nanocomposites with desired properties. To this end, this work has employed atomistic molecular dynamics (MD) simulations to study the thermal properties and interfacial characteristics of TPU composites filled with pristine or functionalized single-walled carbon nanotubes (SWNTs). Simulations reveal that the introduction of SWNTs suppresses TPU chain dynamics and favors the hydrogen bond formation induced by the wrapping of TPU chains around SWNTs, leading to an increase of glass transition temperature (Tg) and a reduction of volumetric coefficient of thermal expansion (CTE) in the rubbery state. Compared to pristine and hydrogenated SWNTs, SWNTs featuring polar groups, such as carboxyl (-COOH), oxhydryl (-OH) and amine (-NH2) groups, show improved affinity for TPU molecules, suppressing polymer mobility. Analysis of SWNT/TPU binding energy and solubility parameter suggests that electrostatic interactions are responsible for such a functionalized SWNT/TPU interface enhancement. Furthermore, the amine groups exhibit the highest potential for forming hydrogen bonds with the urethane carbonyl (-C[double bond, length as m-dash]O) of TPU chains, resulting in lowest polymer mobility and highest Tg. In general, this research work could provide some guidance for material design of polymer nanocomposites and future simulations relevant to TPUs.


The clinical outcome of total knee arthroplasty is compromised by a previously implanted medial unicondylar knee arthroplasty.
To investigate the clinical outcome of patients that underwent conversion of a medial unicondylar knee arthroplasty (UKA) to a total knee arthroplasty (TKA) and to compare that outcome to patients that underwent primary TKA. It was hypothesized that those groups would significantly differ in terms of knee score outcome and implant survival. A retrospective-comparative study was conducted utilizing data from the Federal state's arthroplasty registry. Included were patients from our department that undergone a conversion of a medial UKA to a TKA (UKA-TKA group). The Western Ontario and MacMaster Universities Osteoarthritis Index (WOMAC) from preoperative and 1-year postoperative was used. Moreover, the implant survival was analyzed. In the UKA-TKA group, there were 51 cases (age 67 ± 10, 74% women), and in the TKA group, there were 2247 cases (age 69 ± 9, 66% women). The one-year postoperative WOMAC total score was 33 in the UKA-TKA group und 21 in the TKA group (p < 0.001). Similarly, the WOMAC pain, WOMAC stiffness, and WOMAC function scores were significantly worse in the UKA-TKA. After 5 years, the survival rates were 82% and 95% (p = 0.001). The 10-years prosthesis survival was 74% and 91% in the UKA-TKA and TKA groups, respectively (p < 0.001). Based on our findings it is concluded that patients who received a TKA after UKA have inferior results than those that directly receive a TKA. This is true for both patient-reported knee outcome and prosthesis survival. Converting UKA to TKA should not be seen as an easy operation, but should rather be done by surgeons with considerable experience in both primary and revision knee arthroplasty.


Effect of ultrasound-guided thoracic paravertebral block on perioperative analgesia in elderly patients undergoing video-assisted thoracic lobectomy in China: An interventional clinical randomized controlled trial.
The aim of this study was to investigate the analgesic effect and safety of ultrasound-guided thoracic paravertebral block (UG-TPVB) in Chinese elderly patients undergoing video-assisted thoracic lobectomy (VATL) and to study the influence of aging factors on these effects. This study was a single-center, single-blind, prospective, randomized, controlled trial. A total of 300 patients scheduled for VATL were recruited and randomly divided into the UG-TPVB group (T group) and conventional anesthesia group (C group) according to the recruitment order, and subgroups were set up according to whether the age was ≥65 years old or not. The postoperative 12, 24, and 48 h static/dynamic visual analog scale (VAS) scores, intraoperative fentanyl consumption, postoperative extubation time, post-anesthesia care unit (PACU) stay time, hospitalization days, postoperative complications, and other indicators were compared between the two groups. The postoperative 12, 24, and 48 h static/dynamic VAS scores of the T group were significantly lower than those of the C group. The intraoperative fentanyl consumption, postoperative extubation time, PACU stay time, and postoperative hospitalization days were significantly lower than those of the C group. The incidence of postoperative 48 h urinary retention in the T group was significantly lower than that in the C group. These advantages showed no significant difference or slight difference between elderly patients and nonelderly patients, indicating that UG-TPVB did not influence the analgesic effect and safety of VATL patients by age or age difference. UG-TPVB is an effective and safe perioperative analgesia method for elderly VATL patients. Its application improves the quality of life and prognosis of elderly VATL patients.


IOPA: I/O-aware parallelism adaption for parallel programs.
With the development of multi-/many-core processors, applications need to be written as parallel programs to improve execution efficiency. For data-intensive applications that use multiple threads to read/write files simultaneously, an I/O sub-system can easily become a bottleneck when too many of these types of threads exist; on the contrary, too few threads will cause insufficient resource utilization and hurt performance. Therefore, programmers must pay much attention to parallelism control to find the appropriate number of I/O threads for an application. This paper proposes a parallelism control mechanism named IOPA that can adjust the parallelism of applications to adapt to the I/O capability of a system and balance computing resources and I/O bandwidth. The programming interface of IOPA is also provided to programmers to simplify parallel programming. IOPA is evaluated using multiple applications with both solid state and hard disk drives. The results show that the parallel applications using IOPA can achieve higher efficiency than those with a fixed number of threads.


Medicare minus choice: the impact of HMO withdrawals on rural Medicare beneficiaries.
A disproportionate share of the Medicare beneficiaries who lost coverage as a result of recent health maintenance organization (HMO) withdrawals have been from rural areas. Rural beneficiaries are less likely than urban beneficiaries are to have another Medicare+Choice (M+C) option. We surveyed a nationwide random sample of 1,093 rural beneficiaries to assess the impact of HMO withdrawals. A high proportion of beneficiaries ended up without any coverage beyond traditional Medicare; on average, beneficiaries experienced significant increases in premiums; and the proportion of beneficiaries with prescription drug coverage decreased significantly. These results raise questions about whether the federal government should encourage plans to enter rural markets where they will be the only M+C plan and where their withdrawal could have negative consequences for enrollees who lose coverage.


What are the implications of myomas on fertility? A need for a debate?
In order to evaluate the relationship between leiomyomas and infertility, which remains a subject of debate, we have tried to evaluate the impact of myomas on fertility and pregnancy outcome in different conditions where myomas are implicated. Medline research was conducted of publications appearing between January 1988 and August 2001 on the subjects of myomas and myomectomy and their impact on fertility and pregnancy outcome in infertile women. A total of 106 manuscripts were consulted. The incidence of myomas in infertile women without any obvious cause of infertility is estimated to be 1-2.4%. The relationship between leiomyomas and infertility remains a subject of debate. The question is: do myomas influence fertility? We are obliged to conclude that the question remains. The absence of an answer to this crucial question is probably due to the fact that we have not yet conducted the appropriate prospective studies required to obtain any clear results.


Prevalence of Comorbidities in Asthma and Nonasthma Patients: A Meta-analysis.
This study compares the prevalence rates of comorbidities between asthma and nonasthma control patients reported in the literature.Literature was searched in several electronic databases. After the selection of studies by following précised eligibility criteria, meta-analyses of odds ratios were carried out with subgroup and sensitivity analyses.Eleven studies studying 117,548 asthma patients compared with 443,948 non-asthma controls were included in the meta-analysis. The prevalence of cardiovascular comorbidities (odds ratio (OR): [95% CI] 1.90 [1.70, 2.14]; P < 0.00001), cerebrovascular comorbidities (OR 1.44 [1.29, 1.60]; P < 0.00001), obesity (OR 1.51 [1.14, 2.01]; P < 0.00001), hypertension (OR 1.66 [1.47, 1.88]; P < 0.00001, diabetes (OR 1.25 [1.08, 1.44]; P < 0.00001), other metabolic and endocrine comorbidities (OR 1.60 [1.40, 1.83]; P < 0.00001), psychiatric and neurological comorbidities (OR 1.62 [1.44, 1.82]; P < 0.00001), gut and urinary comorbidities (OR 1.91 [1.47, 2.49]; P < 0.00001),), cancer (OR 1.17 [1.10, 1.25]; P < 0.00001), and respiratory comorbidities (OR 5.60 [4.22, 7.44]; P < 0.00001) were significantly higher in the asthma patients in comparison with nonasthma controls.Asthma is associated with significantly higher comorbidities including cardio-/cerebrovascular diseases, obesity, hypertension, diabetes, psychiatric and neurological comorbidities, gut and urinary conditions, cancer, and respiratory problems other than asthma. Respiratory comorbidities are found 5 times more prevalent in asthma than in non-asthma patients.


Family matters in Canada: understanding and addressing family homelessness in Ontario.
Homelessness is becoming an international public health issue in most developed countries, including Canada. Homelessness is regarded as both political and socioeconomic problems warranting broad and consistent result-oriented approaches. This paper represents the qualitative findings of a project that explored risk factors associated with family homelessness and strategies that could mitigate and prevent homelessness among families using a focused ethnographic study guided by the principles of participatory action research (PAR). The sample includes 36 family members residing at a family shelter who participated in focus groups over two years (between April 2016 and December 2017). Most of the participants were single-parent women. The analysis yielded five major themes including, life challenges, lack of understanding of the system, existing power differentials, escaping from hardship, and a theme of proposed solutions for reducing family homelessness in the community. The findings illustrated the complex nature of family homelessness in Ontario; that the interaction of multiple systems can put families at risk of homelessness. Findings from this study underscore the need for urgent housing protocols aimed at educating homeless families on how to navigate and understand the system, enhance their conflict resolution skills, and develop strategies beyond relocation to help them to cope with difficulties with housing.


The utility of H2 receptor antagonists in preventing infusion-related reactions to paclitaxel chemotherapy.
Paclitaxel has a risk of infusion-related reactions (IRRs) and despite no prospective evidence, is often given with premedication including a corticosteroid, H1 antagonist, and H2 antagonist (H2RA). Backorders impacted the supply of intravenous H2RAs at our center, and it was removed as routine premedication. The authors compared the incidence of IRR in patients treated without H2RA to patients receiving standard H2RA premedication. The authors reviewed outpatients starting paclitaxel at the Ottawa Hospital from December 2019 to October 2021. Two cohorts were created: patients treated without H2RA premedication (intervention), and those receiving standard H2RA (control). Demographics, treatment, and IRR information were collected retrospectively. Primary end point was rate of grade ≥2 IRRs during first two doses of paclitaxel. A total of 182 patients were treated without H2RA premedication, compared to 184 control patients treated during non-backorder periods. Baseline characteristics included: median age, 63 years; 86% female; and primary tumor 52% breast/24% gynecologic/10% gastric/esophageal/8% lung/6% other. There were no significant differences between cohorts in baseline characteristics. There was no difference in the rate of grade ≥2 IRR between cohorts; 12.1% (22 of 182; 95% confidence interval [CI], 7.7%-17.7%) for patients treated without H2RA, and 15.1% (28 of 185; 95% CI, 10.3%-21.1%) for control patients. The rate of grade ≥3 IRRs were also similar, 4.4% in intervention cohort versus 3.8% in control cohort. The removal of H2RAs from premedication for paclitaxel did not result in an increased incidence of IRRs. The use of H2RAs in preventing IRRs to paclitaxel should be re-evaluated.


Identification of a novel EXT1 mutation in patients with hereditary multiple exostosis by exome sequencing.
Hereditary multiple exostosis (HME) is an autosomal inherited skeletal disease whose etiology is not fully understood. To further understand the genetic spectrum of the disease, we analyzed a five-generation Chinese family with HME that have observable inheritance. Exome sequencing was performed on three HME individuals and three unaffected individuals from the family. A downstream study confirmed a new C deletion at codon 442 on exon 5 of the exostosin-1 (EXT1) gene as the only pathogenic site which generated a stop codon and caused the truncation of the protein. We rediscovered the deletion in other affected individuals but not in the unaffected individuals from the family. Upon immunohistochemistry assay, we found that the EXT1 protein level of the patients with the novel mutation in our study was less than the level in the patients without the EXT1 mutation from another unrelated family. For a deeper understanding, we analyzed the mutation spectrum of the EXT1 gene. The present study should facilitate a further understanding of HME.


Antioxidants inhibit ATP-sensitive potassium channels in cerebral arterioles.
Hydrogen peroxide and peroxynitrite are capable of generating hydroxyl radical and are commonly suspected as sources of this radical in tissues. It would be useful to distinguish the source of hydroxyl radical in pathophysiological conditions and to clarify the mechanisms by which antioxidants modify vascular actions of oxidants. We investigated the effect of three antioxidants--dimethylsulfoxide (DMSO), salicylate, and L-cysteine--on the cerebral arteriolar dilation caused by topical application of hydrogen peroxide and peroxynitrite in anesthetized cats equipped with cranial windows. We also tested the effect of these antioxidants on the vasodilation caused by pinacidil and cromakalim, two known openers of ATP-sensitive potassium channels. DMSO was more effective in inhibiting dilation from hydrogen peroxide, whereas salicylate and L-cysteine were more effective in inhibiting dilation from peroxynitrite. All three antioxidants inhibited dilation in concentrations that were remarkably low (< 1 mmol/L). All three antioxidants inhibited vasodilation from two known potassium channel openers, pinacidil and cromakalim. Their effect was specific because they did not affect dilation from adenosine or nitroprusside. The findings show that antioxidants block ATP-sensitive potassium channels in cerebral arterioles. This appears to be the mechanism by which antioxidants inhibit the dilation from hydrogen peroxide and peroxynitrite and not through scavenging of a common intermediate, ie, hydroxyl radical. The differences between effectiveness in inhibiting dilation from hydrogen peroxide and peroxynitrite by various antioxidants suggest that hydrogen peroxide and peroxynitrite act at two different sites, one in a water-soluble environment and the other in a lipid-soluble environment.


General Procedures for the Lithiation/Trapping of N-Boc Piperazines.
To provide α-substituted piperazines for early stage medicinal chemistry studies, a simple, general synthetic approach is required. Here, we report the development of two general and simple procedures for the racemic lithiation/trapping of N-Boc piperazines. Optimum lithiation times were determined using in situ IR spectroscopy, and the previous complicated and diverse literature procedures were simplified. Subsequent trapping with electrophiles delivered a wide range of α-functionalized N-Boc piperazines. The scope and limitations of the distal N-group were investigated. The selective α- and β- arylation of N-Boc piperazines via lithiation/Negishi coupling is reported.


Experiences and views of married women about domestic violence.
The aim of this study was to determine the experiences and views of married women about the topic of domestic violence. This research was planned as a mixed methods study with an in-depth interview and descriptive approach. The study was conducted between November 2011 and December 2012 with 24 married women living in Ankara, Turkey. Two main data-collection tools were used in the study: the "Personal Information Form" and the "In-depth Interview Questionnaire." Data of this study were evaluated by content analysis. A majority of the participants (83.3%) stated that they had been exposed to domestic violence that had been committed primarily by their husbands. The actual reasons for the violence were reported to be such factors as "financial problems and lack of education and love and respect between the couples." It was determined that as the victims became more desperate, they turned to reading of the Koran, prayer, and smoking. Domestic violence adversely affects the physical and mental health of individuals, families, and the entire community. Therefore, it will take a community effort to address the causes of domestic violence and to create viable solutions that will improve the health of everyone.