Gastric Bypass Surgery Reduces De Novo Cases of Type 2 Diabetes to Population Levels: A Nationwide Cohort Study From Sweden.
The aim of this study was to determine long-term changes in pharmacological treatment of type 2 diabetes after primary Roux-en-Y gastric bypass (RYGB) surgery, in patients with and without pharmacological treatment of diabetes preoperatively. Several studies have shown that gastric bypass has good effect on diabetes, at least in the short-term. This study is a nationwide cohort study using Swedish registers, with basically no patients lost to follow-up during up to 7 years after surgery. The effect of RYGB on type 2 diabetes drug treatment was evaluated in this nationwide matched cohort study. Participants were 22,047 adults with BMI ≥30 identified in the nationwide Scandinavian Surgical Obesity Registry, who underwent primary RYGB between 2007 and 2012. For each individual, up to 10 general population comparators were matched on birth year, sex, and place of residence. Prescription data were retrieved from the nationwide Swedish Prescribed Drug Register through September 2015. Incident use of pharmacological treatment was analyzed using Cox regression. Sixty-seven percent of patients with pharmacological treatment of type 2 diabetes before surgery were not using diabetes drugs 2 years after surgery and 61% of patients were not pharmacologically treated up to 7 years after surgery. In patients not using diabetes drugs at baseline, there were 189 new cases of pharmacological treatment of type 2 diabetes in the surgery group and 2319 in the matched general population comparators during a median follow-up of 4.6 years (incidence: 21.4 vs 27.9 per 10,000 person-years; adjusted hazard ratio 0.77, 95% confidence interval 0.67-0.89; P < 0.001). Gastric bypass surgery not only induces remission of pharmacological treatment of type 2 diabetes but also protects from new onset of pharmacological diabetes treatment. The effect seems to persist in most, but not all, patients over 7 years of follow-up.


Chromosomes and causation of human cancer and leukemia. XLV. Chromosome patterns in stimulated lymphocytes of chronic lymphocytic leukemia.
In the present study the B-cell mitogens from Epstein-Barr virus (EBV) and lipopolysaccharide-B from E. coli (LPS) were used to stimulate the blood cells of 27 patients with chronic lymphocytic leukemia (CLL) to undergo mitosis. In 14 CLL cases EBV was used as the only mitotic stimulant and in 5 cases both EBV and LPS. In 8 cases neither EBV nor LPS caused mitotic cells to appear. In 9 cases the cells were characterized by abnormal clones, including 5 cases with +12 (including 1 case with a possible Philadelphia chromosome), 2 cases with 14q+ and 1 case with an i(17q). No correlation was found between the types of surface membrane immunoglobulins and the chromosome abnormalities in the leukemic lymphocytes. The effects of EBV on normal lymphocytes were also investigated and found to be nonspecific (e.g., tetraploidy), although in every case EBV caused a definite increase in the mitotic index. The results indicate that 1) EBV and LPS stimulate leukemic cells of CLL to undergo mitosis, 2) about 50% of the patients have clonal abnormalities in the leukemic cells, and 3) CLL is associated with karyotypic changes seen in lymphoma (14q+), although the most common anomaly in our material was a +12.


PPAR delta agonists stimulate oligodendrocyte differentiation in tissue culture.
Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors of the nuclear hormone receptor superfamily that have been described as master genes that switch cells from an undifferentiated phenotype to a differentiated phenotype. In the present investigation, we examined the possibility that ligands for PPARs are potent activators of oligodendrocyte (OL) differentiation and/or proliferation. Primary glial cultures and enriched OL cultures of neonatal mouse cerebra were treated with three different PPAR agonists: a PPAR gamma-selective agonist, a PPAR delta-selective agonist, and a pan agonist selective for both PPAR gamma and delta. Treatment with PPAR gamma agonist does not have an effect on the differentiation of OLs; however, PPAR delta agonist and the pan agonist treatment accelerates the differentiation of OLs within 24 h of application in mixed glial cultures. The number of OLs with processes and huge membrane sheets increases two- to threefold in both groups. The increase in the size of the sheets is also mirrored by changes in the intensity and distribution of myelin basic protein (MBP) and proteolipid protein (PLP) mRNAs. As compared to controls, the PPAR delta agonist-treated groups contain more OLs that have MBP and PLP mRNA extending into distal processes. These results indicate that PPAR delta plays a significant role in the maturation of OLs and regulates the size of OL sheets. BrdU immunostaining reveals that these agonists do not significantly stimulate proliferation of OLs expressing glycolipids. The studies in enriched OL cultures reproduce the effects of the PPAR agonists seen in the mixed glial cultures, indicating that the effect of the PPAR agonists is directly on the OLs and not via astrocytes. In the enriched cultures, the total number of OLs increases significantly in the PPAR delta agonist-treated groups, but BrdU immunostaining does not show an increased proliferation of cells. These findings suggest that PPAR delta increases the survival of cells and/or prevents cell death in enriched cultures. Although PPAR delta is expressed in various cell types, its role as a factor in the transcriptional regulation of OL differentiation has not been explored. We show for the first time that a ligand that serves as an agonist for PPAR delta activates the program of OL differentiation in primary and enriched OL cultures.


Effect of forskolin on primary cultures of astrocytes and oligodendrocytes.
Forskolin, a unique diterpene which activates adenylate cyclase, stimulated production of cyclic AMP in primary cultures of astrocytes and oligodendrocytes prepared from the cerebral cortex of neonatal rats. At low concentration, forskolin in also potentiated the action of agents (e.g. isoproterenol, prostaglandin E1) which are known to elicit accumulations of cyclic AMP in these cultures. Agents that can stimulate the adenylate cyclase system in astrocyte cultures are inhibited by an alpha-adrenergic agonist, and this alpha-adrenergic mediated inhibition was not observed in oligodendrocyte cultures. Results from this study indicate that forskolin in activation of adenylate cyclase is susceptible to inhibition by an alpha-adrenergic agonist in astrocyte cultures but not in oligodendrocyte cultures. Phenylephrine, which was thought to be acting as a pure alpha-agonist on glial cells, was shown to have both alpha- and beta-adrenergic activities in the same system.


Fine-scale mapping of 8q24 locus identifies multiple independent risk variants for breast cancer.
Previous genome-wide association studies among women of European ancestry identified two independent breast cancer susceptibility loci represented by single nucleotide polymorphisms (SNPs) rs13281615 and rs11780156 at 8q24. A fine-mapping study across 2.06 Mb (chr8:127,561,724-129,624,067, hg19) in 55,540 breast cancer cases and 51,168 controls within the Breast Cancer Association Consortium was conducted. Three additional independent association signals in women of European ancestry, represented by rs35961416 (OR = 0.95, 95% CI = 0.93-0.97, conditional p = 5.8 × 10(-6) ), rs7815245 (OR = 0.94, 95% CI = 0.91-0.96, conditional p = 1.1 × 10(-6) ) and rs2033101 (OR = 1.05, 95% CI = 1.02-1.07, conditional p = 1.1 × 10(-4) ) were found. Integrative analysis using functional genomic data from the Roadmap Epigenomics, the Encyclopedia of DNA Elements project, the Cancer Genome Atlas and other public resources implied that SNPs rs7815245 in Signal 3, and rs1121948 in Signal 5 (in linkage disequilibrium with rs11780156, r(2) = 0.77), were putatively functional variants for two of the five independent association signals. The results highlighted multiple 8q24 variants associated with breast cancer susceptibility in women of European ancestry.


Effect of oxidative DNA damage in promoter elements on transcription factor binding.
Reactive oxygen species produced by endogenous metabolic activity and exposure to a multitude of exogenous agents impact cells in a variety of ways. The DNA base damage 8-oxodeoxyguanosine (8-oxodG) is a prominent indicator of oxidative stress and has been well-characterized as a premutagenic lesion in mammalian cells and putative initiator of the carcinogenic process. Commensurate with the recent interest in epigenetic pathways of cancer causation we investigated how 8-oxodG alters the interaction between cis elements located on gene promoters and sequence-specific DNA binding proteins associated with these promoters. Consensus binding sequences for the transcription factors AP-1, NF-kappaB and Sp1 were modified site-specifically at guanine residues and electrophoretic mobility shift assays were performed to assess DNA-protein interactions. Our results indicate that whereas a single 8-oxodG was sufficient to inhibit transcription factor binding to AP-1 and Sp1 sequences it had no effect on binding to NF-kappaB, regardless of its position. We conclude from these data that minor alterations in base composition at a crucial position within some, but not all, promoter elements have the ability to disrupt transcription factor binding. The lack of inhibition by damaged NF-kappaB sequences suggests that DNA-protein contact sites may not be as determinative for stable p50 binding to this promoter as other, as yet undefined, structural parameters.


Drug resistance and apoptosis in cancer treatment: development of new apoptosis-inducing agents active in drug resistant malignancies.
Modulation of multidrug resistance (MDR) has been extensively studied in vitro and in vivo. However, several clinical trials have failed to show any important benefits in terms of response to chemotherapy or the length of survival using MDR reversing agents. This may be due to the expression or co-expression of other drug resistance mechanisms in malignant cells. Several studies have shown that most, if not all, chemotherapeutic agents exert their anticancer activity by inducing apoptosis; therefore, resistance to apoptosis may be a major factor limiting the effectiveness of anticancer therapy. In the last few years, effort has been made to understand the biochemical alterations of apoptotic pathways in cancer. Many of these alterations confer a multidrug resistant phenotype to malignant cells. In this context, the new recently developed anticancer therapies based on drugs that modulate apoptosis may have importance for the treatment of tumors that are scarcely responsive to the conventional anticancer chemotherapy. In this review, we discuss the current knowledge about drug resistance, apoptosis and cancer and report the recently developed apoptosis modulating strategies that have potential therapeutic implications for the drug resistant tumors.


Directly observed therapy of sofosbuvir/ribavirin +/- peginterferon with minimal monitoring for the treatment of chronic hepatitis C in people with a history of drug use in Chennai, India (C-DOT).
We assessed the feasibility of field-based directly observed therapy (DOT) with minimal monitoring to deliver HCV treatment to people with a history of drug use in Chennai, India. Fifty participants were randomized 1:1 to sofosbuvir+peginterferon alfa 2a+ribavirin (SOF+PR) for 12 weeks (Arm 1) vs sofosbuvir+ribavirin (SOF+R) for 24 weeks (Arm 2). SOF+R was delivered daily at participant chosen venues and weekly peginterferon injections at the study clinic. HCV RNA testing was performed to confirm active HCV infection and sustained virologic response 12 weeks after treatment completion (SVR12). No baseline genotyping or on-treatment viral loads were performed. Median age was 46 years. All were male and 20% had significant fibrosis/cirrhosis. All self-reported history of injection drug use, 18% recent noninjection drug use and 38% alcohol dependence. Six discontinued treatment (88% completed treatment in each arm). Of 22 who completed SOF+PR, all achieved SVR12 (22/25=88%); 15 of 22 who completed SOF+R achieved SVR12 (15/25=60%; P=.05). Among those completing SOF+R, SVR12 was significantly less common in participants reporting ongoing substance use (36% vs 100%) and missed doses. Active substance use and missed doses did not impact SVR with SOF+PR. Field-based DOT of HCV therapy without real-time HCV RNA monitoring was feasible; however, achieving 100% adherence was challenging. SOF+PR appeared superior to SOF+R in achieving SVR12, even when doses were missed with no discontinuations due to side effects. Further exploration of short duration treatment with peginterferon plus direct-acting antivirals is warranted.


Developing an integrated model for outcomes management.
In the complexity of integrated delivery systems, the triad of clinicians, financial analysts, and information management personnel can partner to form an effective force in driving outcomes management efforts. Health care systems can take the opportunity to rethink outcomes management strategies with an emphasis on monitoring cost and quality simultaneously. Advanced practice nurses, with a wide breadth of knowledge obtained through higher education, are positioned to be active, effective team members at all points in the outcomes management process.


A method for the direct injection and analysis of small volume human blood spots and plasma extracts containing high concentrations of organic solvents using revered-phase 2D UPLC/MS.
The emergence of micro sampling techniques holds great potential to improve pharmacokinetic data quality, reduce animal usage, and save costs in safety assessment studies. The analysis of these samples presents new challenges for bioanalytical scientists, both in terms of sample processing and analytical sensitivity. The use of two dimensional LC/MS with, at-column-dilution for the direct analysis of highly organic extracts prepared from biological fluids such as dried blood spots and plasma is demonstrated. This technique negated the need to dry down and reconstitute, or dilute samples with water/aqueous buffer solutions, prior to injection onto a reversed-phase LC system. A mixture of model drugs, including bromhexine, triprolidine, enrofloxacin, and procaine were used to test the feasibility of the method. Finally an LC/MS assay for the probe pharmaceutical rosuvastatin was developed from dried blood spots and protein-precipitated plasma. The assays showed acceptable recovery, accuracy and precision according to US FDA guidelines. The resulting analytical method showed an increase in assay sensitivity of up to forty fold as compared to conventional methods by maximizing the amount loaded onto the system and the MS response for the probe pharmaceutical rosuvastatin from small volume samples.


Genetic Diversity in Natural Populations of the Near-Threatened Species Lignobrycon myersi (Characiformes, Triportheidae): Implications for Species Conservation.
The river basins of Brazil contain a highly diverse ichthyofauna of remarkable endemism, including several threatened species. Accordingly, Lignobrycon myersi is a fish species distributed only in a few rivers from the state of Bahia, northeastern Brazil. Since this species is classified as Near Threatened and is poorly studied, efforts to understand the genetic structure of populations and putative cryptic forms should help define efficient strategies of management and conservation. Herein, the molecular identification and the population genetic diversity of specimens of L. myersi across their range (Almada, Contas, and Cachoeira river basins) were assessed using mitochondrial markers (16S rDNA and D-Loop, respectively). The inferences based on phylogenetics, genetic distance, and species delimitation methods invariably identified all samples as L. myersi. In addition, sequencing of D-loop fragments revealed significant haplotype diversity and a considerable level of population genetic structure. Despite their geographic isolation, these data suggested that populations from Almada and Contas rivers represent a single evolutionary lineage that could be managed as a whole. In contrast, the population from Cachoeira River was highly differentiated from the others and should be managed separately as a unique and endemic unit, particularly focused on the conservation of native habitats.


Positive contrast magnetic resonance imaging of cells labeled with magnetic nanoparticles.
Contrast agents incorporating superparamagnetic iron-oxide nanoparticles have shown promise as a means to visualize labeled cells using MRI. Labeled cells cause significant signal dephasing due to the magnetic field inhomogeneity induced in water molecules near the cell. With the resulting signal void as the means for detection, the particles behave as a negative contrast agent, which can suffer from partial-volume effects. In this paper, a new method is described for imaging labeled cells with positive contrast. Spectrally selective RF pulses are used to excite and refocus the off-resonance water surrounding the labeled cells so that only the fluid and tissue immediately adjacent to the labeled cells are visible in the image. Phantom, in vitro, and in vivo experiments show the feasibility of the new method. A significant linear correlation (r = 0.87, P < 0.005) between the estimated number of cells and the signal was observed.


Real-world evidence of cabozantinib in patients with metastatic renal cell carcinoma: Results from the CABOREAL Early Access Program.
Real-world data on cabozantinib in metastatic renal cell carcinoma (mRCC) is limited. This study (CABOREAL) reports treatment patterns and outcomes for patients treated with cabozantinib through the French Early Access Program. This multicentre (n = 26), observational, retrospective study enrolled patients with mRCC who had received ≥1 dose of cabozantinib. Overall survival (OS) was estimated using the Kaplan-Meier method; subgroups were compared using the log-rank test. A multiple Cox regression model assessed predictive factors of OS after cabozantinib initiation. Four hundred and ten recruited patients started treatment between September 2016 and February 2018: the Eastern Cooperative Oncology Group Performance Status ≥2, 39.3%; poor International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk, 31.7%; 0-1, 2 and ≥3 previous treatment lines, 25.3%, 33.4% and 41.2%, respectively; bone metastases, 55.9%; brain metastases, 16.8%. Median (min-max) follow-up was 14.4 (0-30) months. Overall, 57.0% of patients had a dose reduction, 15.6% an alternative dose schedule. The median average daily dose was 40.0 mg. Median (quartile [Q]1-Q3) treatment duration was 7.6 (0.1-29.1) months, median OS was 14.4 months, and the 12-month OS rate was 56.5% (95% confidence interval: 51.5-61.2). Most patients (54.4%) received subsequent treatment. Predictive factors associated with longer OS were body mass index ≥25 kg/m2 (p = 0.0021), prior nephrectomy (p = 0.0109), favourable or intermediate IMDC risk (p < 0.0001) and cabozantinib initiation at 60 mg/day (p = 0.0486). In the largest real-world study to date, cabozantinib was effective in unselected, heavily pretreated patients with mRCC. Initiation at 60 mg/day was associated with improved outcomes. CLINICALTRIALS. NCT03744585.


Changes in the expression of α-tocopherol-related genes in liver and mammary gland biopsy specimens of peripartum dairy cows.
Blood α-tocopherol (α-Toc) concentrations decline gradually throughout the prepartum period, reaching the nadir after calving in dairy cows. The 6 α-Toc-related molecules [α-Toc transfer protein (TTPA); afamin; scavenger receptor class B, Type I; ATP-binding cassette transporter A1; tocopherol-associated protein (SEC14L2); and cytochrome P450 family 4, subfamily F, polypeptide 2 (CYP4F2)] are expressed in liver and other peripheral tissues. These molecules could regulate α-Toc transport, blood concentrations, and metabolism of α-Toc. Therefore, the aim of this study was to evaluate the changes in the expression of α-Toc-related genes in liver and mammary gland tissues of dairy cows around calving, which have remained elusive until now. In experiment (Exp.) 1, 28 multiparous Holstein cows were used (from -5 to 6 wk relative to parturition) to monitor the changes in dietary α-Toc intake, blood concentrations of α-Toc, and lipoproteins; in Exp. 2, 7 peripartum Holstein cows were used (from -4 to 4 wk relative to parturition) for liver tissue biopsy; and in Exp. 3, 10 peripartum Holstein cows were used (from -8 to 6 wk relative to parturition) to carry out the mammary gland tissue biopsy and milk sampling. In Exp. 1, the serum α-Toc concentrations declined gradually with decreasing amount of α-Toc intake and plasma high-density lipoprotein concentrations toward calving time. However, in the early lactation period after calving, serum α-Toc concentrations remained at a lower concentration despite the recovery of α-Toc intake and plasma high-density lipoprotein concentrations. In Exp. 2, just after calving, the TTPA, SEC14L2, afamin, and albumin mRNA expression levels in the liver were temporarily downregulated, and the hepatic mRNA levels of endoplasmic reticulum stress-induced unfolded protein response markers and acute-phase response marker increased at calving. In Exp. 3, the concentrations of α-Toc in colostrum were greater than those in precolostrum (samples were collected at wk -1 relative to parturition) and mature milk. The expression of TTPA, SEC14L2, and CYP4F2 mRNA in bovine mammary gland tissue was detected. However, TTPA and SEC14L2 mRNA expressions showed the opposite trends: the expression levels of TTPA mRNA peaked whereas SEC14L2 mRNA reached a nadir at calving. These results indicate that the expression of α-Toc-related genes involved in specific α-Toc transfer and metabolism in the liver and mammary gland are altered during calving. Moreover, these changes might be associated with the maintenance of lower serum α-Toc concentrations after calving.


TRPM2, calcium and neurodegenerative diseases.
NMDA receptor overactivation triggers intracellular Ca(2+) dysregulation, which has long been thought to be critical for initiating excitotoxic cell death cascades associated with stroke and neurodegenerative disease. The inability of NMDA receptor antagonists to afford neuroprotection in clinical stroke trials has led to a re-evaluation of excitotoxic models of cell death and has focused research efforts towards identifying additional Ca(2+) influx pathways. Recent studies indicate that TRPM2, a member of the TRPM subfamily of Ca(2+)-permeant, non-selective cation channel, plays an important role in mediating cellular responses to a wide range of stimuli that, under certain situations, can induce cell death. These include reactive oxygen and nitrogen species, tumour necrosis factor as well as soluble oli-gomers of amyloid beta. However, the molecular basis of TRPM2 channel involvement in these processes is not fully understood. In this review, we summarize recent studies about the regulation of TRPM2, its interaction with calcium and the possible implications for neurodegenerative diseases.


Surgical Treatment of Rhinogenic Contact Point Headache: An Experience from a Tertiary Care Hospital.
Introduction Even in the absence of inflammatory disease, facial pain often results from pressure of two opposing nasal mucosa surfaces. Objectives The objective of this study is to assess the efficacy of surgical treatment of contact point headache. Methods Our study enrolled patients with unilateral facial pain and without nasal/paranasal sinus disease. We confirmed the presence of mucosal contact by nasal endoscopy and by computed tomography. Forty-two subjects with the three most common anatomical variations underwent complete evaluation: 17 with concha bullosa (CB), 11 with septal deviation (SD), and 14 with septal spur (SS). All participants were treated by topical corticosteroid, adrenomimetic, and antihistamine. The patients without improvement were treated surgically. We assessed the severity of pain using a Visual Analogue Score (VAS) before surgical treatment and one, six, twelve, and twenty-four months after. Results The patients with SS had more severe facial pain in comparison with patients with CB (p = 0.049) and SD (p = 0.000). The subjects with CB had higher degree of facial pain than the ones with SD (p = 0.001). After an unsuccessful medical treatment and surgical removal of mucosal contacts, the decrease of headache severity was more intense in patients with CB and SS (p = 0.000) than in the patients with SD (p = 0.01). Conclusion Our results suggest that topical medications have no effects and that surgical removal of mucosal contacts could be effective in the treatment of contact point headache. The results of surgical treatment were better in cases of facial pain caused by SS and CB, than in those caused by SD.


How Can Home Care Patients and Their Caregivers Better Manage Fall Risks by Leveraging Information Technology?
From the perspectives of home care patients and caregivers, this study aimed to (a) identify the challenges for better fall-risk management during home care episodes and (b) explore the opportunities for them to leverage health information technology (IT) solutions to improve fall-risk management during home care episodes. Twelve in-depth semistructured interviews with the patients and caregivers were conducted within a descriptive single case study design in 1 home health agency (HHA) in the mid-Atlantic region of the United States. Patients and caregivers faced challenges to manage fall risks such as unmanaged expectations, deteriorating cognitive abilities, and poor care coordination between the HHA and physician practices. Opportunities to leverage health IT solutions included patient portals, telehealth, and medication reminder apps on smartphones. Effectively leveraging health IT could further empower patients and caregivers to reduce fall risks by acquiring the necessary information and following clinical advice and recommendations. The HHAs could improve the quality of care by adopting IT solutions that show more promise of improving the experiences of patients and caregivers in fall-risk management.


Exohedral Functionalization of Fullerene by Substituents Controlling of Molecular Organization for Spontaneous C60 Dimerization in Liquid Crystal Solutions and in a Bulk Controlled by a Potential.
A study was carried out on the possibility of orderly and spontaneous dimerization at room temperature of C60 cages in fullerene liquid crystal fullerene dyads (R-C60). For this purpose, dyads with a structural elements feature supporting π-stacking and Van der Waals interactions were tested, due to the presence of terthiophene donors linked through an α-position or dodecyloxy chains. In addition, this possibility was also tested and compared to dyads with shorter substituents and the pristine C60. Research has shown that only in dyads with the features of liquid crystals, π-dimerization of C60 units occurs, which was verified by electrochemical and spectroelectrochemical (ESR) measurements. Cyclic voltammetry and differential voltammetry studies reveal π-dimerization in liquid crystal dyad solution even without the possibility of previous polymerization (cathodic or anodic) under conditions in the absence of irradiation and without the availability of reaction initiators, and even with the use of preliminary homogenization. These dyads undergo six sequential, one-electron reductions of π-dimer (R-C60···C60-R), where two electrons are added successively to each of the two fullerene cages and first form two radical anion system (R-C60)•-(R-C60)•- without pairing with the characteristics of two doublets. Similarly, the second reductions of π-dimer occur at potentials that are close to the reduction potential for the conversion to a system of two triplet dianions (R-C60)2-(R-C60)2-. Electron paramagnetic resonance spectra indicate a significant interaction between C60 cages. Interestingly, the strength of intermolecular bonds is so significant that it can overcome Coulombic repulsion, even with such highly charged particles as dianions and trianions. Such behavior has been revealed and studied so far only in covalently bonded C60 dimers.


Improved outcome of coronary stenting by utilizing a modified treatment protocol.
Coronary stents improve patient outcome, are easy to deploy and are now being utilized very frequently after coronary angioplasty. New protocols have been suggested to overcome the main limitations of stenting, namely subacute stent thrombosis, local vascular complications, and the length of hospitalization. In this report we describe our current results with coronary stenting in 100 patients: we modified our standard protocol and used high pressure noncompliant balloons to expand the stent, added ticlopidine to some of the patients to inhibit platelet activity, and used the transradial approach in some patients to reduce the risk of vascular complications. These results were compared to those obtained in our initial group of 50 patients in whom we used a standard implantation protocol. Subacute thrombosis occurred in one patient in the current group of patients compared to five patients in our early experience (P = 0.026). There was no change in local vascular complications. This preliminary report shows that subacute thrombosis can almost be eliminated by the new stent implantation protocols. Less aggressive anticoagulation and radial artery catheterization can decrease vascular complications.


Nanosized ethosomal dispersions for enhanced transdermal delivery of Nebivolol using intradermal/transfollicular sustained reservoir: in vitro evaluation, confocal laser scanning microscopy and in vivo pharmacokinetic studies.
Nebivolol (NBV), a BCS class-II anti-hypertensive drug that suffers from limited solubility and oral bioavailability. Nanosized ethosomes were adopted as an approach to solubilize and deliver NBV transdermally, as a substitute to oral route. Ethosomal dispersions were prepared employing thin film hydration method. Formulation variables were adjusted to obtain entrapment efficiency; EE > 50%, particle size; PS < 100 nm, zeta potential; ZP > ± 25 mV and polydispersity index; PDI < 0.5. The optimized ethosomal dispersion (OED) showed accepted EE 86.46 ± 0.15%, PS 73.50 ± 0.08 nm, ZP 33.75 ± 1.20 mV, and PDI 0.31 ± 0.07. It also showed enhanced cumulative amount of NBV permeated at 8h (Q8) 71.26 ± 1.46%, and 24h (Q24) 98.18 ± 1.02%. TEM images denoted spherical vesicles with light colored lipid bilayer and dark core. Confocal laser scanning microscopy showed deeply localized intradermal and transfollicular permeation of the fluorolabelled OED (FL-OED). Nanosized FL-OED (< 100 nm) can permeate through hair follicles creating a drug reservoir for enhanced systemic absorption. OED formulated into transdermal patch (OED-TP1) exhibited accepted physicochemical properties including; thickness 0.14 ± 0.01 mm, folding endurance 151 ± 0.07, surface pH 5.80 ± 0.15, drug content 98.64 ± 2.01%, mucoadhesion 8534 ± 0.03, Q8 87.61 ± 0.11% and Q24 99.22 ± 0.24%. In vivo pharmacokinetic studies showed significantly enhanced bioavailability of OED-TP1 by 7.9 folds compared to oral Nevilob® tablets (p = 0.0002). It could be concluded that OED-TP1 can be a promising lipid nanocarrier TDDS for NBV and an efficacious alternative route of administration for hypertensive patients suffering from dysphagia.


Alteration in membrane fluidity and lipid composition, and modulation of H(+)-ATPase activity in Saccharomyces cerevisiae caused by decanoic acid.
Decanoic acid, a lipophilic agent, inhibited in vitro the plasma membrane H(+)-ATPase of Saccharomyces cerevisiae grown in YPD medium. Conversely, when decanoic acid (35 microM) was present in the growth medium, the measured H(+)-ATPase activity was four times higher than that of control cells. Km, and pH and orthovanadate sensitivity were the same for the two growth conditions, which indicated that H(+)-ATPase activation was not due to conformational changes in the enzyme. The activation process was not entirely reversible which showed that plasma membrane H(+)-ATPase activation is due to several mechanisms. 1,6-diphenyl-1,3,5-hexatriene anisotropy performed on protoplasts from cells grown in YPD revealed that as decanoic acid concentration was increased, anisotropy significantly decreased, i.e. membrance fluidity increased. Cells grown in media containing decanoic acid exhibited greater membrane fluidity compared with control cells. Furthermore, these cells did not show any fluidifying effect when increased concentrations of decanoic acid were added. Chemical analysis of cell membrane lipid composition revealed a modification in the distribution of the phospholipid fatty acids and sterols in cells grown in the presence of 35 microM decanoic acid compared with control cells. Our results support the view that the plasma membrane H(+)-ATPase activation induced by decanoic acid is correlated with an alteration in membrane lipid constituents.


Propylthiouracil blocks extrathyroidal conversion of thyroxine to triiodothyronine and augments thyrotropin secretion in man.
Propylthiouracil (PTU) inhibits peripheral deiodination of thyroxine (T4) and triiodothyronine (T3) and decreases the metabolic effectiveness of T4 in animals. To assess the effect of PTU on extrathyroidal conversion of T4 to T3 in man, 15 studies were performed in athyreotic patients treated with 100 or 200 mug of L-T4 daily for 1 mo before the addition of PTU, 250 mg every 6 h for 8 days. serum T3, T4, and thyrotropin (TSH) were measured daily by radioimmunoassay; serum TSH response to 500-mug thyrotropin-releasing hormone (TRH) was measured before and on the last day of giving PTU. On the 100-mug LT4 dose, serum T3 fell from 120 plus or minus 5 (SE) to 83 plus or minus 6 ng/dl (P less than 0.005) with return to 113 plus or minus 5 ng/dl after stopping PTU; serum T4 (4.5 plus or minus 0.3 mug/dl) did not change. Similar results were seen in patients taking 200 mug of L-T4 daily. On the 100-mug dose of L-T4 the fall in T3 was accompanied by a reciprocal rise in serum TSH to 195 plus or minus 33% of initial concentration (P less than 0.01) with return to 104 plus or minus 8% after PTU. The serum TSH response to TRH (DELTAMUU/ml over base line) was greater during PTU therapy than during the control period. On 100-mug L-T4 DELTA TSH rose from 64 plus or minus 19 to 101 plus or minus 23 muU/ml (P less than 0.005). Expressed as percent of base-line TSH concentration, TSH rose from 140 plus or minus 52 to 280 plus or minus 44% (control vs. PTU) at 15 min, 265 plus or minus 72 to 367 plus or minus 63% at 30 min, 223 plus or minus 54 to 313 plus or minus 54% at 45 min, 187 plus or minus 45 to 287 plus or minus 51% at 60 min, and 145 plus or minus 22 to 210 plus or minus 28% at 120 min after TRH. The data suggest that PTU blocks extrathyroidal conversion of T4 to T3, thus increasing pituitary TSH secretion and augmenting the TSH response to TRH.


Diagnosis ex juvantibus. Individual response patterns to drugs reveal hypertension mechanisms and simplify treatment.
Heterogeneity of response to antihypertensive therapy is a well-recognized clinical phenomenon. An agent that is antihypertensive in one patient may increase blood pressure in another or have no effect in a third. We believe that this variety of individual response to drug treatment can provide a new framework for the study of hypertensive subjects. Different patterns of response elicited by sequential trials of individual drugs with different mechanisms of action (diuretics, calcium channel blockers, alpha-blockers, beta-blockers, and converting enzyme inhibitors) should provide another means to classify hypertensive patients into biologically relevant groups. The documentation and analysis of this therapeutic heterogeneity in relation to renin profiling and to other physiological and demographic parameters may add a new dimension to the investigation of the pathophysiology of hypertension; it may serve as a basis for more appropriate stratification of participants in clinical trials and may ultimately contribute to a more rational approach to patient management.


[The significance of pesticin 1 for the virulence and immunogenicity of Yersinia pestis].
The work deals with the study of the virulent and immunogenic properties of Y. pestis strains which lost their capacity for producing pesticine 1 as the result of the insertion of a Tn-like element into the 6-MD plasmid responsible for this property. The "switching-off" of gene pst induced a decrease in the virulence of Y. pestis injected subcutaneously into white mice and guinea pigs and had no influence on its level of immunogenicity for white mice. A suggestion was made that pesticine 1 played no essential role in the expression of the virulence and immunogenicity of Y. pestis penetrating into the body by subcutaneous route.


The law of the pack.
The value of the Internet lies in its connections, but some connections are more valuable than others. Which matter most? Reed's Law can tell you.


Split Spy0128 as a potent scaffold for protein cross-linking and immobilization.
Site-specific cross-linking techniques between proteins and additional functional groups have become increasingly important for expanding the utility of proteins in biochemistry and biotechnology. In order to explore powerful techniques for practical bioconjugation applications, we have validated a technique mediated by a unique property of Streptcoccus pyogenes pilin subunit Spy0128, an autocatalytic intramolecular isopeptide formation in Spy0128. Recently, it has been revealed that Spy0128 can be split into two fragments (split-Spy0128 (residues 18-299 of Spy0128) and isopeptag (residues 293-308 of Spy0128)) that were capable of forming an intermolecular covalent complex. We focused on this unique reconstitution property and first studied the bioconjugation of blue and green fluorescent proteins, enabling the direct monitoring of cross-linking reactions by Förster resonance energy transfer (FRET). A fluorescence lifetime study shows that spatial control of two proteins on the Spy0128 scaffold is possible when one protein is fused to the N-terminus of split-Spy0128 and another one is tethered at the N- or C-terminus of the isopeptag. Furthermore, we demonstrated site-specific protein immobilization mediated by the reconstitution of split-Spy0128 and isopeptag. In this case, a split-Spy0128 mutant with a free N-terminal Cys residue was first immobilized onto beads chemically modified with a maleimide group through a Michael addition process. Then, an isopeptagged protein was successfully immobilized onto the split-Spy0128-immobilized beads. These results suggest that Spy0128 is a potent proteinaceous scaffold available for bioconjugation both in solution and at a solid surface.


AdmixPower: Statistical Power and Sample Size Estimation for Mapping Genetic Loci in Admixed Populations.
Admixed populations result from recent admixture of two or more ancestral populations with divergent allele frequencies. The genome of each admixed individual is a mosaic of haplotypes inherited from the ancestral populations. Despite the substantial work to assess power and sample size requirements for association mapping in genetically homogeneous populations of European ancestry, power and sample size estimation methods for mapping genes in genetically heterogeneous admixed populations such as African Americans are lacking. Admixture mapping is a method that traces the ancestral origin of disease-susceptibility genetic loci in the admixed population. We developed AdmixPower, a freely available tool set based on the open-source R software, to perform power and sample size analysis for genetically heterogeneous admixed populations considering continuous or dichotomous outcomes with a case-only or case-control study design. AdmixPower can be used to compute the sample size required to achieve investigator-specified statistical power under several key parameters including ancestry odds ratio, genotype risk ratio, parental risk ratio, an underlying genetic risk model, trait type, and admixture model (hybrid-isolation or continuous gene flow model). We demonstrate that differences in the key parameters in the admixed population results in substantial differences in the sample size required to achieve adequate power in admixture mapping studies. Our tool provides a resource for researchers to develop a strategy to minimize cost and maximize the success of identifying disease-susceptibility loci in an admixed population. R code used in the sample size and power analysis is freely available from https://research.cchmc.org/mershalab/Tools.html.


Does dose matter in reducing gestational weight gain in exercise interventions? A systematic review of literature.
The purpose of this review was to examine the relationship between exercise dose and reductions in weight gain during pregnancy in exercise interventions. Systematic literature review. Four electronic research databases (PubMed, Web of Science, CINAHL, and Academic Search Premiere) were used to identify exercise interventions conducted with pregnant women. Eligible articles must have satisfied the following criteria: inclusion of a control condition, exercise as a major intervention component, weight gain measured and reported for each experimental condition, description of exercise dose (frequency, intensity and duration), and utilized an adequate number of control conditions to assess independent effects of exercise on weight gain. The literature search identified 4837 articles. Of these, 174 abstracts were screened and 21 intervention studies (18 exercise-only, 3 exercise/diet) were eligible for review. Only 38% of the interventions achieved statistically significant reductions in gestational weight gain. Successful interventions possessed higher adherence and lower attrition rates and were predominantly conducted among normal weight populations. No clear patterns or consistencies of exercise dose and reductions in weight gain were evident. An exercise dose associated with reductions in weight gain was unquantifiable among these interventions. Adherence and retention rates were strong contributors to the success of exercise interventions on gestational weight gain. It is strongly suggested that future researchers investigate methods to increase adherence and compliance, especially among overweight and obese women, and utilize objective measurement tools to accurately evaluate exercise dose performed by the participants and the impact on body composition and weight gain.


Complete nucleotide sequence of Iranian tomato yellow leaf curl virus isolate: further evidence for natural recombination amongst begomoviruses. Brief Report.
Complete nucleotide sequence of the Iranian strain of tomato yellow leaf curl virus (TYLCV-IR) was determined and compared with some begomoviruses. The complete sequence of TYLCV-IR clustered together with TYLCV and TYLCV-MId from Israel. A similar relationship holds when the deduced amino acid sequences of V1, V2, C2 and C3 and nucleotide sequences of IR, and RIR were compared. In contrast, phylogenetic analyses of amino acid sequences of C4, C1, and nucleotide sequences of LIR revealed that TYLCV-IR clustered with TLCIRV and two Indian species: ToLCBV- [Ban4], and ToLCKV. The phylogenetic analyses, Recombination Detection Program analyses, and sequence alignment survey provided evidence of the occurrence of recombination between an Israeli TYLCV-MId, as major parent, and TLCIRV, as minor parent. In this recombination event, a region (from nt 2149 to 2766) of TYLCV-MId genome were replaced with corresponding genome sequences of TLCIRV (RDP P-value = 5.976 x 10(-72)), which include LIR, C4, and N-terminal of C1. Infectivity of the cloned TYLCV-IR genome was demonstrated by successful agroinoculation of tomato ( Lycopersicon esculentum) and other plant species. The disease was transmitted by the natural vector Bemisia tabaci from agroinoculated plants to test plants, reproducing in this way the full biological cycle and proving that the genome of TYLCV-IR consists of only one circular single-stranded DNA molecule.


Case Report: Scrambler Therapy for Treatment-Resistant Central Neuropathic Pain in a Patient with Transverse Myelitis.
Central neuropathic pain is a severely disabling consequence of conditions that cause tissue damage in the central nervous system. It is often refractory to treatments commonly used for peripheral neuropathy. Scrambler therapy is an emerging noninvasive pain-modifying technique that uses transcutaneous electrical stimulation of nociceptive fibers with the intent of reorganizing maladaptive signaling pathways. It has been examined for the treatment of peripheral neuropathy with favorable safety and efficacy outcomes, but its application to central neuropathic pain has not been reported in transverse myelitis. We describe the use of Scrambler therapy in a patient with persistent central neuropathic pain due to transverse myelitis. The patient had tried multiple drugs for treatment of the pain, but they were not effective or caused adverse effects. After a course of Scrambler therapy, pain scores improved considerably more than what was reported with previous pharmacologic and nonpharmacologic interventions. This case supports further investigation of Scrambler therapy in multiple sclerosis, neuromyelitis optica spectrum disorder, and other immune-mediated disorders that damage the central nervous system.


Oxygen inhibition of nitrogenase activity in Klebsiella pneumoniae.
A purpose-built oxystat has been used to study reversible inhibition of nitrogenase by O2 in the facultative anaerobe Klebsiella pneumoniae. C2H2-reducing activity in samples from either an anaerobic glucose-limited or an O2-limited diazotrophic chemostat culture was completely inhibited by exposure to a dissolved O2 concentration (DOC) of 1.5 microM or above. Subsequently, under anaerobic conditions, C2H2-reducing activity returned in the absence of de novo protein synthesis. The amount of activity returning never reached 100% of the initial anaerobic activity before O2 treatment. The degree of reversibility was inversely proportional to the log of DOC during exposure and was decreased by increasing the time of exposure to O2 (about 60% reversibility occurred after a 20 min exposure to 6 microM-O2). The failure to obtain complete recovery of activity was apparently not due to inactivation of the very O2-sensitive pyruvate-flavodoxin oxidoreductase (nifJ product) which provides electrons for nitrogenase activity in vivo. Samples from the O2-limited culture behaved similarly to those limited by glucose. Thus, 'training' of the organism to use O2 during growth does not influence the tolerance of nitrogenase to O2. Since the behaviour towards O2 reported here for K. pneumoniae differs from that known to occur in Azotobacter, the mechanism of protection of nitrogenase from O2 damage may differ in these organisms.


Synthesis of 3'alpha-carbo-substituted 2',3'-dideoxyribonucleosides from naturally occurring nucleosides.
Naturally occurring ribonucleosides, uridine and adenosine, were converted to their 3'alpha-CH2CO2Me and 3'alpha-CH2PO(OPh)2 2',3'-dideoxy analogues. The present reaction sequence starts with oxidative cleavage of the 2',3'-cis-diol system, and involves radical-mediated reconstruction of furanose ring as the key step.


Actigraphy to Evaluate Sleep in the Intensive Care Unit. A Systematic Review.
Poor sleep quality is common in the intensive care unit (ICU) and may be associated with adverse outcomes. Hence, ICU-based efforts to promote sleep are gaining attention, motivating interest in methods to measure sleep in critically ill patients. Actigraphy evaluates rest and activity by algorithmically processing gross motor activity data, usually collected by a noninvasive wristwatch-like accelerometer device. In critically ill patients, actigraphy has been used as a surrogate measure of sleep; however, its use has not been systematically reviewed. To conduct a systematic review of ICU-based studies that used actigraphy as a surrogate measure of sleep, including its feasibility, validity, and reliability as a measure of sleep in critically ill patients. We searched PubMed, EMBASE, CINAHL, Proquest, and Web of Science for studies that used actigraphy to evaluate sleep in five or more patients in an ICU setting. Our search yielded 4,869 citations, with 13 studies meeting eligibility criteria. These 13 studies were conducted in 10 countries, and eight (62%) were published since 2008. Across the 13 studies, the mean total sleep time of patients in the ICU, as estimated using actigraphy, ranged from 4.4 to 7.8 hours at nighttime and from 7.1 to 12.1 hours over a 24-hour period, with 1.4 to 49.0 mean nocturnal awakenings and a sleep efficiency of 61 to 75%. When compared side-by-side with other measures of sleep (polysomnography, nurse assessments, and patient questionnaires), actigraphy consistently yielded higher total sleep time and sleep efficiency, fewer nighttime awakenings (vs. polysomnography), and more overall awakenings (vs. nurse assessment and patient questionnaires). None of the studies evaluated the association between actigraphy-based measures of sleep and outcomes of patients in the ICU. In critically ill patients, actigraphy is being used more frequently as a surrogate measure of sleep; however, because actigraphy only measures gross motor activity, its ability to estimate sleep is limited by the processing algorithm used. Prior ICU-based studies involving actigraphy were heterogeneous and lacked data regarding actigraphy-based measures of sleep and patient outcomes. Larger, more rigorous and standardized studies are needed to better understand the role of actigraphy in evaluating sleep and sleep-related outcomes in critically ill patients.


Receptor-mediated metabolism of the phosphoinositides and phosphatidic acid in rat lacrimal acinar cells.
The metabolism of the inositol lipids and phosphatidic acid in rat lacrimal acinar cells was investigated. The muscarinic cholinergic agonist methacholine caused a rapid loss of 15% of [32P]phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] and a rapid increase in [32P]phosphatidic acid (PtdA). Chemical measurements indicated that the changes in 32P labelling of these lipids closely resembled changes in their total cellular content. Chelation of extracellular Ca2+ with excess EGTA caused a significant decrease in the PtdA labelling and an apparent loss of PtdIns(4,5)P2 breakdown. The calcium ionophores A23187 and ionomycin provoked a substantial breakdown of [32P]PtdIns(4,5)P2 and phosphatidylinositol 4-phosphate (PtdIns4P); however, a decrease in [32P]PtdA was also observed. Increases in inositol phosphate, inositol bisphosphate and inositol trisphosphate were observed in methacholine-stimulated cells, and this increase was greatly amplified in the presence of 10 mM-LiCl; alpha-adrenergic stimulation also caused a substantial increase in inositol phosphates. A23187 provoked a much smaller increase in the formation of inositol phosphates than did either methacholine or adrenaline. Experiments with excess extracellular EGTA and with a protocol that eliminates intracellular Ca2+ release indicated that the labelling of inositol phosphates was partially dependent on the presence of extracellular Ca2+ and independent of intracellular Ca2+ mobilization. Thus, in the rat lacrimal gland, there appears to be a rapid phospholipase C-mediated breakdown of PtdIns(4,5)P2 and a synthesis of PtdA, in response to activation of receptors that bring about an increase in intracellular Ca2+. The results are consistent with a role for these lipids early in the stimulus-response pathway of the lacrimal acinar cell.


Targeting cancer with the resources at hand: rolando camacho, MD, national oncology group.
Rolando Camacho was born in Pinar del R�o-Cuba's westernmost province and, ironically enough, home to the country's richest tobacco-producing farmlands. He was about to leave for an engineering scholarship abroad when he accompanied a friend to medical school on the first day of classes in Havana, a bus ride that changed his life. Since completing his oncology residency in 1985, Dr Camacho has served as director of Pinar del R�o's Cancer Control Program, chair of the National Cancer Control Program, director of the National Oncology and Radiobiology Institute (INOR), and today, chair of the National Oncology Group. The Group advises the Ministry of Public Health through its National Cancer Control Unit on questions of cancer control policy, planning and development of human resources for cancer care, structure of services, material resource planning and cancer research. Since 1988, breast cancer has been a particular focus for Dr Camacho's research, practice and teaching. MEDICC Review spoke with Dr. Camacho for his views on the problem of cancer in Cuba, and the prospects for more effective control.


Knowledge, attitudes and practices in tamarind (Tamarindus indica L) use and conservation in Eastern Uganda.
Tamarindus indica L is one of the indigenous fruit tree species that traditionally contributes to food security and ecosystem stability in sub-Saharan Africa. We hypothesized that the indigenous people of Eastern Uganda have used T. indica for generations and developed practices that promote its conservation and therefore we expected that they possess an elaborate indigenous knowledge (IK) system and that most of them have planted the species. The aim of this study was to assess the influence of IK, attitudes and practices on the use and conservation of T. indica. A cross sectional survey was conducted in two districts purposively selected from the T. indica natural range. Focus group discussions, key informant interviews, semi-structured interviews and observation were used to collect data. Data was processed using qualitative analytical methods. Tamarindus indica was highly valued by the majority of the population. Tamarindus indica was used for food, medicinal, cultural, social, environmental amelioration and income generation purposes. The population possessed a high level of IK about T. indica evidenced by 18 categories of uses and multiple modes of use. Fruit pulp was the most commonly used tamarind product. Relative frequency of citation of the different uses provides insight into usage levels and IK possessed. The communities' food and medicinal uses concurred with scientific reports of health benefits of consuming T. indica. Approximately half of respondents had T. indica on their compounds or homegardens (53%). Fifty two percent of the tamarind population was self-propagated, 45% were planted while the propagation history for the remaining 3% was not known. Constraints towards planting T. indica included limited land, long maturation period and low monetary value. Fifty three percent of those who were growing T. indica did not carry out any silvicultural practices. The majority of T. indica encountered (87%) was intercropped with other crops or trees. Several beliefs and taboos regarding T. indica persist. Tamarindus indica has a high use value in the study area evidenced by multiple uses. Indigenous knowledge and uses concurred with scientifically proven nutritional and medicinal attributes of T. indica in literature which is significant given current trends towards affordable functional foods. The high level of IK has not translated into high rates of planting T. indica. There is need to encourage value-addition so as to maximize T. indica benefits and enhance conservation.


[Hearing disorders in Paget's disease].
The audiometric data of 41 patients with Paget's disease of the bone show that hearing loss exceeds the age-related presbycusis in patients with deformities of the temporal bone as demonstrated by X-ray (27%). The usual type of hearing loss is a sensorineural one. 80% of these patients showed hair cell damage, 32% of them a retrocochlear lesion, depending on the degree of deformity of the temporal bone. Conductive hearing loss of the otosclerosis type was found in only 20%. The Paget-related hearing loss can be regarded as a consequence of the deformation occurring both at the bone surrounding the cochlea and at the internal auditory canal.


Assessment of the factors influencing primary care physicians' approach to vaccination of adult risk groups in Istanbul, Turkey.
We aimed to assess the factors influencing primary care physicians' (PCPs) approach to adult vaccination in specific risk groups and evaluate the compliance to adult immunization guidelines. This cross-sectional study performed between January 2016 and April 2016 in İstanbul, Turkey. A questionnaire designed to obtain physicians' demographical data, experience, immunization status, and attitude on prescribing or recommending vaccines for adults in the risk group. Healthy individuals older than 65 and patients suffer from chronic diseases or had splenectomy before are considered as a risk group. The questionnaire was sent via email to a randomly selected group of 1,500 PCPs. The data of 221 physicians who responded emails were recorded for statistical analysis. Of the 221 participants (123 women, 98 men), the majority were aged 31-40 years. Their vaccination rates were 74.2% for hepatitis B, 54.3% for seasonal influenza, and 47.1% for tetanus. Among participants, the highest recommendation and prescription rate of adult vaccines was recorded in PCPs aged 31-40 years. In addition, PCPs with <10 years occupational experience were found to prescribe adult vaccines more frequently than PCPs with longer occupational experience. Primary care physicians with lower age and relatively less experience are more intent to prescribe adult vaccines to patients that are in risk groups. This result may be due to increased awareness of adult immunization among PCPs who had more recent medical training. However, many other factors could have caused this difference, including physicians' approach to primary medical care.


Feminine charm: an experimental analysis of its costs and benefits in negotiations.
The authors examined feminine charm, an impression management technique available to women that combines friendliness with flirtation. They asked whether feminine charm resolves the impression management dilemma facing women who simultaneously pursue task (i.e., economic) and social goals in negotiations. They compared women's social and economic consequences after using feminine charm versus a neutral interaction style. They hypothesized that feminine charm would create positive impressions of its users, thus partially mitigating the social penalties women negotiators often incur. They also expected that the degree to which females were perceived as flirtatious (signaling a concern for self), rather than merely friendly (signaling a concern for other), would predict better economic deals for females. Hypotheses were supported across a correlational study and three experiments. Feminine charm has costs and benefits spanning economic and social measures. Theoretical and practical implications are discussed.


[Drug therapy of spastic disorder of urine outflow in patients with spinal injury and spinal cord injury].
A study was made of efficiency of medicamentous correction of a spastic disorder of outflow of urine from the bladder in 42 patients with dull trauma of the spinal column and marrow. The use in the above patients of cardura alpha 1-adrenoblocker permits restoring a spastic disorder of urine outflow in a 10-day period after the trauma sustained in 82 percent of patients. Shortening the time of a spastic disorder of urine outflow in patients with trauma of the spinal column and cord we allow the frequency of development of the infectious inflammatory process in organs of the urogenitary system to be substantially reduced.


Establishment of Patient-Derived Pancreatic Cancer Organoids from Endoscopic Ultrasound-Guided Fine-Needle Aspiration Biopsies.
Three-dimensional cultures of human pancreatic cancer tissue also known as "organoids" have largely been developed from surgical specimens. Given that most patients present with locally advanced and/or metastatic disease, such organoids are not representative of the majority of patients. Therefore, we used endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) to collect pancreatic cancer tissues from patients with advanced pancreatic cancer to create organoids, and evaluated their utility in pancreatic cancer research. Single-pass EUS-FNA samplings were employed to obtain the tissue for organoid generation. After establishment of the organoid, we compared the core biopsy tissues with organoids using hematoxylin and eosin staining, and performed whole exome sequencing (WES) to detect mutational variants. Furthermore, we compared patient outcome with the organoid drug response to determine the potential utility of the clinical application of such organoid-based assays. Organoids were successfully generated in 14 of 20 tumors (70%) and were able to be passaged greater than 5 times in 12 of 20 tumors (60%). Among them, we selected eight pairs of organoid and core biopsy tissues for detailed analyses. They showed similar patterns in hematoxylin and eosin staining. WES revealed mutations in KRAS, TP53, CDKN2A, SMAD4, BRCA1, and BRCA2 which were 93% homologous, and the mean nonreference discordance rate was 5.47%. We observed moderate drug response correlations between the organoids and clinical outcomes in patients who underwent FOLFIRINOX chemotherapy. The established organoids from EUS-FNA core biopsies can be used for a suitable model system for pancreatic cancer research.


Nanocrystals generated under tensile stress in metallic glasses with phase selectivity.
Revealing the mechanism of phase selectivity can provide guidance for controlling crystals with certain phases for special properties. In the present work, nanocrystals of about 2-4 nm diameters with a B2 structure (thermodynamic metastable phase) are generated from CuZr glassy fiber by applying tensile stress at ambient temperature. By combining the ab initio calculations with the molecular dynamics simulations, the stabilities of B2 austenite and B19' martensitic phases under applied tensile stress are compared, and the phase transformation mechanism is revealed. The results show that the B2 structure has a bigger attractive basin, and the phase transition could occur with a larger applied stress during the deformation. Therefore, insights into the higher symmetric B2 nanocrystal with selective nucleation driven under directional stress are provided.


Adenovirus-mediated overexpression of p15INK4B inhibits human glioma cell growth, induces replicative senescence, and inhibits telomerase activity similarly to p16INK4A.
The genes encoding the cyclin-dependent kinase inhibitors p16INK4A (CDKN2A) and p15INK4B (CDKN2B) are frequently homozygously deleted in a variety of tumor cell lines and primary tumors, including glioblastomas in which 40-50% of primary tumors display homozygous deletions of these two loci. Although the role of p16 as a tumor suppressor has been well documented, it has remained less well studied whether p15 plays a similar growth-suppressing role. Here, we have used replication-defective recombinant adenoviruses to compare the effects of expressing wild-type p16 and p15 in glioma cell lines. After infection, high levels of p16 and p15 were observed in two human glioma cell lines (U251 MG and U373 MG). Both inhibitors were found in complex with CDK4 and CDK6. Expression of p16 and p15 had indistinguishable effects on U251 MG, which has homozygous deletion of CDKN2A and CDKN2B, but a wild-type retinoblastoma (RB) gene. Cells were growth-arrested, showed no increased apoptosis, and displayed a markedly altered cellular morphology and repression of telomerase activity. Transduced cells became enlarged and flattened and expressed senescence-associated beta-galactosidase, thus fulfilling criteria for replicative senescence. In contrast, the growth and morphology of U373 MG, which expresses p16 and p15 endogenously, but undetectable levels of RB protein, were not affected by exogenous overexpression of either inhibitor. Thus, we conclude that overexpression of p15 has a similar ability to inhibit cell proliferation, to cause replicative senescence, and to inhibit telomerase activity as p16 in glioma cells with an intact RB protein pathway.


Sepsis-Induced Immunosuppression in Neonates.
Neonates, especially those born preterm, are at increased risk of sepsis and adverse long-term effects associated with infection-related inflammation. Distinct neonatal immune responses and dysregulated inflammation are central to this unique susceptibility. The traditional separation of sepsis into an initial hyper-inflammatory response followed by hypo-inflammation is continually under review with new developments in this area of research. There is evidence to support the association of mortality in the early acute phase of sepsis with an overwhelming hyper-inflammatory immune response. Emerging evidence from adults suggests that hypo- and hyper-inflammation can occur during any phase of sepsis and that sepsis-immunosuppression is associated with increased mortality, morbidity, and risk to subsequent infection. In adults, sepsis-induced immunosuppression (SII) is characterised by alterations of innate and adaptive immune responses, including, but not limited to, a prominent bias toward anti-inflammatory cytokine secretion, diminished antigen presentation to T cells, and reduced activation and proliferation of T cells. It is unclear if sepsis-immunosuppression also plays a role in the adverse outcomes associated with neonatal sepsis. This review will focus on exploring if key characteristics associated with SII in adults are observed in neonates with sepsis.


Regulation of apoptosis in theXenopus embryo by Bix3.
Members of the Bix family of homeobox-containing genes are expressed in the vegetal hemisphere of the Xenopus embryo at the early gastrula stage. Misexpression of at least some of the family members causes activation of mesoderm- and endoderm-specific genes and it is known that some of the proteins, including Bix2 and Bix3, interact with Smad proteins via a motif that is also present in the related protein Mixer. In this paper we study the function of Bix3. Misexpression of Bix3, similar to misexpression of other members of the Bix family, causes the activation of a range of mesendodermal genes, but the spectrum of genes induced by Bix3 differs from that induced by Bix1. More significantly, we find that overexpression of Bix3 also causes apoptosis, as does depletion of Bix3 by use of antisense morpholino oligonucleotides. The ability of Bix3 to causes apoptosis is not associated with its ability to activate transcription and nor with its possession of a Smad interaction motif. Rather, Bix3 lacks a C-terminal motif, which, in Bix1, acts in cis to inhibit apoptosis. Mutation of this sequence in Bix1 causes the protein to acquire apoptosis-inducing activity.


Understanding arsenate reaction kinetics with ferric hydroxides.
Understanding arsenic reactions with ferric hydroxides is important in understanding arsenic transport in the environment and in designing systems for removing arsenic from potable water. Many experimental studies have shown that the kinetics of arsenic adsorption on ferric hydroxides is biphasic, where a fraction of the arsenic adsorption occurs on a time scale of seconds while full equilibrium may require weeks to attain. This research employed density functional theory modeling in order to understand the mechanisms contributing to biphasic arsenic adsorption kinetics. The reaction energies and activation barriers for three modes of arsenate adsorption to ferric hydroxides were calculated. Gibbs free energies of reaction depended on the net charge of the complexes, which is a function of the system pH value. Physical adsorption of arsenate to ferric hydroxide proceeded with no activation barrier, with Gibbs free energies of reaction ranging from -21 to -58 kJ/mol. The highest Gibbs free energies of reaction for physical adsorption resulted from negative charge assisted hydrogen bonding between H atoms on the ferric hydroxide and O atoms in arsenate. The conversion of physically adsorbed arsenate into monodentate surface complexes had Gibbs free energies of activation ranging from 62 to 73 kJ/mol, and Gibbs free energies of reaction ranging from -23 to -38 kJ/mol. The conversion of monodentate surface complexes to bidentate, binuclear complexes had Gibbs free energies of activation ranging from 79 to 112 kJ/mol and Gibbs free energies of reaction ranging from -11 to -55 kJ/mol. For release of arsenate from uncharged bidentate complexes, energies of activation as high as 167 kJ/mol were encountered. Increasingly negative charges on the complexes lowered the activation barriers for desorption of arsenate, and in complexes with -2 charges, the highest activation barrier was 65 kJ/mol. This study shows that the slow kinetics associated with arsenic adsorption and desorption can be attributed to the high Gibbs free energies of activation for forming and breaking bonds with the ferric hydroxide.


A Social Network Family-Focused Intervention to Promote Smoking Cessation in Chinese and Vietnamese American Male Smokers: A Feasibility Study.
Smoking prevalence is high among limited English-proficient Chinese and Vietnamese American men, who are frequently unmotivated to quit and who underutilize smoking cessation resources. This study applied lay health worker outreach to leverage peer and family networks to promote smoking cessation among these men. We integrated qualitative formative research findings and Social Network Theory to develop a social-network family-focused intervention. In a pilot single-group trial, 15 lay health workers recruited 96 dyads (N = 192, 75% Vietnamese) of Chinese or Vietnamese male daily smokers and their family members and delivered the intervention consisting of two small group education sessions and two individual telephone calls over 2 months. At baseline, 42% of smokers were at precontemplation. At 3 months following the initiation of the intervention, 7-day and 30-day point prevalence smoking abstinence rates as reported by smokers and independently corroborated by family members were 30% and 24%, respectively. Utilization of smoking cessation resources (medication, quitline, physician's advice) increased from 2% to 60% (P < .001). Findings showed high acceptability of the intervention as it facilitated learning about tobacco-related health risks and cessation resources, and communications between smokers and their families. This novel social network family-focused intervention to promote smoking cessation among Chinese and Vietnamese smokers appears to be acceptable, feasible, and potentially efficacious. Findings warrant evaluation of long-term efficacy of the intervention in a larger scale randomized controlled trial.


Tumor heterogeneity and resistance to EGFR-targeted therapy in advanced nonsmall cell lung cancer: challenges and perspectives.
Lung cancer, mostly nonsmall cell lung cancer, continues to be the leading cause of cancer-related death worldwide. With the development of tyrosine kinase inhibitors that selectively target lung cancer-related epidermal growth factor receptor mutations, management of advanced nonsmall cell lung cancer has been greatly transformed. Improvements in progression-free survival and life quality of the patients were observed in numerous clinical studies. However, overall survival is not prolonged because of later-acquired drug resistance. Recent studies reveal a heterogeneous subclonal architecture of lung cancer, so it is speculated that the tumor may rapidly adapt to environmental changes via a Darwinian selection mechanism. In this review, we aim to provide an overview of both spatial and temporal tumor heterogeneity as potential mechanisms underlying epidermal growth factor receptor tyrosine kinase inhibitor resistance in nonsmall cell lung cancer and summarize the possible origins of tumor heterogeneity covering theories of cancer stem cells and clonal evolution, as well as genomic instability and epigenetic aberrations in lung cancer. Moreover, investigational measures that overcome heterogeneity-associated drug resistance and new assays to improve tumor assessment are also discussed.


Loose stools lead to fecal incontinence among gynecological cancer survivors.
Many patients treated with radiotherapy to the pelvic region report a change in bowel habits. Loose stools, urgency and fecal incontinence may have a significant impact on daily life and social functioning. We attempted to follow up 789 women, treated with pelvic radiotherapy for a gynecological cancer during 1991 to 2003 at two departments of gynecological oncology in Sweden. A control group of 478 women from the Swedish Population Registry was also included. As a preparatory study, we made in-depth interviews with 26 women previously treated for gynecological cancer. Based on their narratives, we constructed a study-specific questionnaire including 351 questions and validated it face-to-face. The questionnaire covered questions of physical symptoms originating in the pelvis, demographics, psychological and quality of life factors. In relation to bowel symptoms, 60 questions were asked. Six-hundred and sixteen (78%) gynecological cancer survivors and 344 (72%) control women participated. Two-hundred and twenty-six (37%) cancer survivors reported loose stools at least once a week. Eighty-three percent of the survivors with loose stools every day reported defecation urgency with fecal leakage, compared to 20% of cancer survivors without loose stools. Cancer survivors with loose stools at least once a week were 7.7 times more likely to suffer from defecation urgency with fecal leakage (95% CI 4.4-13.3) compared to those who had loose stools once a month or less. In order to avoid loose stools affected survivors with loose stools often skipped meals (13%), made an active choice of food (47%) and preferentially used prescribed medication (36%). There is a relation between loose stools and defecation urgency with fecal leakage among long-term gynecological cancer survivors treated with pelvic radiotherapy. Targeting loose stools can possibly help survivors to decrease frequency of fecal leakage.


FISH Identifies Chromosome Differentiation Between Contemporary Genomes of Wild Types and the Ancestral Genome of Unisexual Clones of Dojo Loach, Misgurnus anguillicaudatus.
In dojo loach (Misgurnus anguillicaudatus), although most wild types are gonochoristic diploids that are genetically differentiated into 2 groups, A and B, clonal lineages appear in certain localities. Clonal loaches have been considered to have hybrid origins between the 2 groups by a series of genetic studies. In this study, using FISH with a newly developed probe (ManDra-A), we identified 26 (1 pair of metacentric and 12 pairs of telocentric chromosomes) of 50 diploid chromosomes in contemporary wild-type group A loach. In contrast, ManDra-A signals were not detected on metacentric chromosomes derived from the ancestral group A of clonal loach. The FISH results clearly showed the presence of certain differentiations in metacentric chromosomes between ancestral and contemporary group A loach. Two-color FISH with ManDra-A and group B-specific ManDra (renamed ManDra-B) probes reconfirmed the hybrid origin of clones by identifying chromosomes from both groups A and B in metaphases. Our results showed the hybrid origin of clonally reproducing fish and the possibility that chromosomal differentiation between ancestral and contemporary fish can affect gametogenesis. In meiotic spermatocytes of sex-reversed clones, ManDra-A, and not ManDra-B, signals were detected in 12 out of 50 bivalents. Thus, the results further support the previous conclusion that clonal gametogenesis was assured by pairing between sister chromosomes duplicated from each ancestral chromosome from group A or B. Our study deepens the knowledge about the association between clonality and hybridity in unisexual vertebrates.


Patient preferences for the treatment of type 2 diabetes: a scoping review.
As more studies report on patient preferences, techniques are needed to identify, assess and, eventually, synthesize results from a diverse set of methodologies. Data on patient preferences are valuable to decision makers in a variety of ways. Preferences for outcomes can be used to inform decision and cost-effectiveness models, while preferences for treatments can inform patient-centered outcomes research (PCOR) and patient-centered care. This project sought to identify and assess the literature reporting on the treatment preferences of adult patients with type 2 diabetes. In addition to cataloging the preference elicitation methods used, we developed and assessed a novel quality assessment checklist for preference-based studies. PubMed, EMBASE, CINAHL, and EconLit databases were searched to identify studies examining patient preferences for medications for type 2 diabetes studies published since inception of each database. The review protocol specified inclusion of studies reporting diabetes-treatment preferences among adults with type 2 diabetes, using a range of preference measurement methods. Studies were excluded if participants were not patients with type 2 diabetes and if treatments were not pharmacological therapies targeting glycemic control, or if no primary preference information was collected. Two investigators independently reviewed titles, abstracts, and articles sequentially to select studies for data abstraction based on the inclusion and exclusion criteria. Disagreements were resolved by consensus. Data on study country, year, number of respondents, preference elicitation method, number of attributes, subgroup analyses, and funding source were abstracted into standardized tables. A novel checklist (PREFS) was used to assess the data quality and validity across different types of preference studies by assessing the following: purpose of the study; respondent sampling; explanation of preference assessment methods; findings reported for total sample; and significance testing. Each item was scored, and an aggregate score was then calculated (ranging from 0 to 5). Of the 2,100 unique citations, 61 met the inclusion criteria. The studies used conjoint analysis (n = 10), time trade-off (n = 6), standard gamble (n = 2), contingent valuation (n = 1), other stated preference methods (n = 39), and revealed preferences (n = 5). Sample sizes ranged from 27 to 14,033, with an average of 562 respondents, and two-thirds included a subgroup analysis. Most studies were conducted in one country, predominantly the USA (n = 27), UK (n = 14), Canada (n = 10), and Germany (n = 7), while 14 were conducted in multiple (2-18) countries across two or more countries. There was an increase in the annual rate of studies published over time from the time of the first publication in 1985 (p = < 0.01). Most (n = 52) studies were funded by pharmaceutical or device companies, with government, academic, association, and hospital sources also funding studies. One study met all five of the PREFS criteria and 12 met four; yet four studies met none of the criteria. The average was 27. Currently, preferences reviews are limited by the mixed quality in the reporting of studies, the publication bias inherent in the literature, a lack of guidelines to conduct various methods, and the difficulty of synthesizing results from different studies. Our study is also limited by its focus on English language articles. This study provides the first systematic evaluation of the methods used in the broad existing body of research into patient preferences for type 2 diabetes medications and can serve as a primary source of information for decision makers. Future work is necessary to assess the utility of the results of reviews of preference information and to develop best-practice guidelines for the reporting of, and methods of conducting, preference studies and systematic reviews of such studies. This systematic review was registered with PROSPERO (registration number CRD42012002285).


Neurogenic chronic idiopathic intestinal pseudo-obstruction, patent ductus arteriosus, and thrombocytopenia segregating as an X linked recessive disorder.
We present a family with three affected males in two generations with congenital neurogenic chronic idiopathic intestinal pseudo-obstruction (CIIP), patent ductus arteriosus, and large platelet thrombocytopenia apparently segregating as an X linked recessive disorder. The pattern of segregation of DNA markers within the family is consistent with linkage to the previously described neurogenic CIIP (CIIPX) locus at Xq28. This combination may represent a new contiguous gene disorder and appears to have a good prognosis with supportive therapy.


Comparison of three hand dynamometers in relation to the accuracy and precision of the measurements.
Given the variety of available hand-held dynamometers and their different handle shapes, reliability studies are needed. To compare the accuracy and reliability between three different hand-held dynamometers and analyze the influence of their handles on grip strength. The tests were performed with the Jamar® dynamometer, the Takei® dynamometer and the EMG System Manual Transducer with modified handle. Eighteen healthy volunteers aged 20.0±1.3 years without history of musculoskeletal disorders or trauma in the evaluated limbs were included. Data normality was tested using the Shapiro-Wilk test. To verify possible differences between the dynamometers, repeated measures ANOVA was administered, followed by Tukey post-hoc tests. Reliability between measurements was evaluated using intraclass correlation coefficient (ICC) and agreement was tested using Bland and Altman plots. The dynamometers calibration process was evaluated using linear regressions. We observed statistically significant differences on the female group between the Jamar® and the Takei® dynamometers (females p<0.001 and males p=0.022) and the EMG System Manual Transducer (female p<0.001 and males p=0.007). However, the Takei® dymamometer and the EMG System Manual Transducer were similar for both female (p=0.161) and male groups (p=0.850). Although acceptable values of intraclass correlation coefficients between measurements were identified, low agreement between the Jamar® dynamometer and all other instruments was found. The results demonstrated that there is an influence of the dynamometer's handle shapes on the measurements of grip strength. Furthermore, the results demonstrated the need for previous calibration of this type of instrument.


Lateral prefrontal cortex: architectonic and functional organization.
A comparison of the architecture of the human prefrontal cortex with that of the macaque monkey showed a very similar architectonic organization in these two primate species. There is no doubt that the prefrontal cortical areas of the human brain have undergone considerable development, but it is equally clear that the basic architectonic organization is the same in the two species. Thus, a comparative approach to the study of the functional organization of the primate prefrontal cortex is more likely to reveal the essential aspects of the various complex control processes that are the domain of frontal function. The lateral frontal cortex appears to be functionally organized along both a rostral-caudal axis and a dorsal-ventral axis. The most caudal frontal region, the motor region on the precentral gyrus, is involved in fine motor control and direct sensorimotor mappings, whereas the caudal lateral prefrontal region is involved in higher order control processes that regulate the selection among multiple competing responses and stimuli based on conditional operations. Further rostrally, the mid-lateral prefrontal region plays an even more abstract role in cognitive control. The mid-lateral prefrontal region is itself organized along a dorsal-ventral axis of organization, with the mid-dorsolateral prefrontal cortex being involved in the monitoring of information in working memory and the mid-ventrolateral prefrontal region being involved in active judgments on information held in posterior cortical association regions that are necessary for active retrieval and encoding of information.


Angiotensin-I converting enzyme inhibitors as potential anti-angiogenic agents for cancer therapy.
Angiotensin-I converting enzyme inhibitors (ACE-Is) are commonly used as safe antihypertensive agents, and it has recently been suggested that they decrease the risk of cancer development. Recent studies have revealed that the renin-angiotensin system (RAS) is involved in the development of many types of tumor. Angiotensin-II (AT-II) has many biological effects, including neo-vascularization, which plays a pivotal role in tumor development. AT-II induces a potent angiogenic factor, namely the vascular endothelial growth factor (VEGF). Some studies have proven that several ACE-Is are potent inhibitors of experimental tumor development and angiogenesis at clinically comparable doses. VEGF expression in tumors is also significantly suppressed by ACE-Is. When used in combination with the conventional anti-cancer drugs, ACE-Is exert more potent anti-tumor activities as compared with either single agent, in addition to suppression of the intra-tumoral angiogenesis. Furthermore, ACE-Is reportedly not only suppress tumor growth but also attenuate the carcinogenesis process in which angiogenesis is involved. Since ACE-Is are already in widespread clinical case without any serious adverse effects, they may represent a potential new strategy for cancer therapy and chemoprevention.


Consolidated BRCA1/2 Variant Interpretation by MH BRCA Correlates with Predicted PARP Inhibitor Efficacy Association by MH Guide.
BRCA1/2 variants are prognostic biomarkers for hereditary breast and/or ovarian cancer (HBOC) syndrome and predictive biomarkers for PARP inhibition. In this study, we benchmarked the classification of BRCA1/2 variants from patients with HBOC-related cancer using MH BRCA, a novel computational technology that combines the ACMG guidelines with expert-curated variant annotations. Evaluation of BRCA1/2 variants (n = 1040) taken from four HBOC studies showed strong concordance within the pathogenic (98.1%) subset. Comparison of MH BRCA's ACMG classification to ClinVar submitter content from ENIGMA, the international consortium of investigators on the clinical significance of BRCA1/2 variants, the ARUP laboratories, a clinical testing lab of the University of UTAH, and the German Cancer Consortium showed 99.98% concordance (4975 out of 4976 variants) in the pathogenic subset. In our patient cohort, refinement of patients with variants of unknown significance reduced the uncertainty of cancer-predisposing syndromes by 64.7% and identified three cases with potential family risk to HBOC due to a likely pathogenic variant BRCA1 p.V1653L (NM_007294.3:c.4957G > T; rs80357261). To assess whether classification results predict PARP inhibitor efficacy, contextualization with functional impact information on DNA repair activity were performed, using MH Guide. We found a strong correlation between treatment efficacy association and MH BRCA classifications. Importantly, low efficacy to PARP inhibition was predicted in 3.95% of pathogenic variants from four examined HBOC studies and our patient cohort, indicating the clinical relevance of the consolidated variant interpretation.


Three-dimensional porous microarray of gold modified electrode for ultrasensitive and simultaneous assay of various cancer biomarkers.
A three-dimensional (3D) microarray of a gold modified electrode with a nanoporous surface was successfully fabricated in this work. The special gold micro/nanostructure possessed an extremely high surface area (ca. 20 times its geometrical area), as well as highly stable and easily chemically modified properties, which could distinctly increase the binding sites for biological probes, facilitate the diffusion profile and enhance the electron transfer. Owing to these advantages, an ultrasensitive biosensor was designed on the porous microarray of the gold modified electrode (PMGE), which realized the simultaneous assay of cancer biomarkers of angiogenin (Ang) and thrombin (Tob). Under optimal experimental conditions, an impressively ultralow detection limit of 0.07 pM for Ang (a linear range from 0.2 pM to 10 nM) and a limit of 20 fM for Tob (a linear range from 50 fM to 5 nM) were obtained. Besides, the fabricated biosensor showed an excellent stability, good reproducibility and a high selectivity towards other biological proteins, which also exhibited promising potential for the application in a real serum sample analysis. Such a micro/nanostructure of gold could have widespread applications in biological sensing and quantitative biochemical analysis.


Clinicopathologic features and treatment outcomes in Woringer-Kolopp disease.
Woringer-Kolopp disease, also known as pagetoid reticulosis, is an exceedingly rare variant of mycosis fungoides. Accurate diagnosis and effective treatment is essential to prevent progression to debilitating disease. We identified 7 patients with Woringer-Kolopp disease treated at our institution. We review the major clinical and pathologic characteristics of this disease, focusing on treatment strategies and patient outcomes. All of our patients were successfully treated with skin-directed therapies including topical steroids, topical nitrogen mustard, psoralen plus ultraviolet A, narrow-band ultraviolet B, and radiation therapy. Our observations confirm that Woringer-Kolopp disease carries an excellent prognosis, and support that the most effective and appropriate treatment for recalcitrant or severe Woringer-Kolopp disease is localized radiation therapy.


Functional Magnetic Resonance Imaging in Abstinent MDMA Users: A Review.
Ecstasy or 3,4-methylenedioxymethamphetamine (MDMA) is a popular drug of abuse. In the animal studies MDMA has been shown to have deleterious effects on the serotonergic neurotransmitter system. Understanding the adverse effects of MDMA on human brain function is of considerable importance owing to the rising number of MDMA users. Various neuroimaging studies have investigated the structural, chemical and functional differences in the brain integrity of chronic MDMA users. Various neurocognitive domains like working memory, episodic memory, semantic memory, visual stimulation, motor function and impulsivity have been compared between chronic MDMA users and nonusers using fMRI. The fMRI studies remain much more sensitive in studying the neurological deficits associated with chronic MDMA use as compared to the cognitive studies alone and therefore they serve as a prelude in our understanding of MDMA induced neurotoxicity. However they still face certain limitations contributing to inconsistency in the results and further research is needed before we can draw definitive conclusions regarding the neurotoxic effects of MDMA.


Combining PGSE NMR with homonuclear dipolar decoupling.
A new robust approach for combining multiple-pulse homonuclear decoupling and PGSE NMR is introduced for accurately measuring molecular diffusion coefficients in systems with nonvanishing static homonuclear dipolar couplings. Homonuclear decoupling suppresses dipolar dephasing during the gradient pulses but its efficiency and scaling factor for the effective gradient vary across the sample because of the large variation of the frequency offset caused by the gradient. The resulting artifacts are reduced by introducing a slice selection scheme. The method is demonstrated by (19)F PGSE NMR experiments in a lyotropic liquid crystal.


Exposure to heavy metals from point pollution sources and risk of incident type 2 diabetes among women: a prospective cohort analysis.
Heavy metal exposures may contribute to diabetes risk but prospective studies are uncommon. We analyzed the Australian Longitudinal Study on Women's Health (three cohorts aged 18-23, 45-50, or 70-75 at baseline in 1996, N = 34,191) merged with emissions data for 10 heavy metals (As, Be, Co, Cr, Cu, Hg, Mn, Ni, Pb, Zn) from the National Pollutant Inventory. Over 20-year follow-up, 2,584 women (7.6%) reported incident diabetes. Cox proportional hazards regression models showed that women aged 45-50 at baseline had higher diabetes risk in association with exposure to total air emissions, total water emissions, all individual metals air emissions, and six individual water emissions. After correction for false discovery rate, nine of 11 air emissions and five water emissions remained significant. Associations were not observed for land-based emissions, or for younger or older cohorts. Emissions were dominated by mining, electricity generation and other metals-related industrial processes.


A melting method for RNA extraction from the mucosal membrane of the mouse middle ear.
There is much confusion surrounding the methods of RNA extraction from the middle ear mucosa of mice. In this study, we worked to develop a "melting method," which is faster, purer, and more reliable than other methods in common use. Thirty-two ears were used for this study. Light microscopy with hematoxylin-eosin staining of the bullae, scanning electron microscopy (SEM), spectrophotometer analysis, and reverse transcription polymerase chain reaction were performed before and after melting the half lateral bullae, which were detached from the temporal bone by using a lateral retroauricular approach. Each resected half bulla contained a well distributed mucosal membrane. After a TRIzol melting duration of 10-30 minutes, only mucosal marker (MUC5AC) was expressed without bony marker (total osteocalcin). The same results were determined from SEM. This melting method, compared with stripping and irrigation methods, is effective and offers an easier, more robust approach to extracting RNA from the middle ear mucosal membranes of mice.


Computational pharmacogenotype extraction from clinical next-generation sequencing.
Next-generation sequencing (NGS), including whole genome sequencing (WGS) and whole exome sequencing (WES), is increasingly being used for clinic care. While NGS data have the potential to be repurposed to support clinical pharmacogenomics (PGx), current computational approaches have not been widely validated using clinical data. In this study, we assessed the accuracy of the Aldy computational method to extract PGx genotypes from WGS and WES data for 14 and 13 major pharmacogenes, respectively. Germline DNA was isolated from whole blood samples collected for 264 patients seen at our institutional molecular solid tumor board. DNA was used for panel-based genotyping within our institutional Clinical Laboratory Improvement Amendments- (CLIA-) certified PGx laboratory. DNA was also sent to other CLIA-certified commercial laboratories for clinical WGS or WES. Aldy v3.3 and v4.4 were used to extract PGx genotypes from these NGS data, and results were compared to the panel-based genotyping reference standard that contained 45 star allele-defining variants within CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, CYP4F2, DPYD, G6PD, NUDT15, SLCO1B1, TPMT, and VKORC1. Mean WGS read depth was >30x for all variant regions except for G6PD (average read depth was 29 reads), and mean WES read depth was >30x for all variant regions. For 94 patients with WGS, Aldy v3.3 diplotype calls were concordant with those from the genotyping reference standard in 99.5% of cases when excluding diplotypes with additional major star alleles not tested by targeted genotyping, ambiguous phasing, and CYP2D6 hybrid alleles. Aldy v3.3 identified 15 additional clinically actionable star alleles not covered by genotyping within CYP2B6, CYP2C19, DPYD, SLCO1B1, and NUDT15. Within the WGS cohort, Aldy v4.4 diplotype calls were concordant with those from genotyping in 99.7% of cases. When excluding patients with CYP2D6 copy number variation, all Aldy v4.4 diplotype calls except for one CYP3A4 diplotype call were concordant with genotyping for 161 patients in the WES cohort. Aldy v3.3 and v4.4 called diplotypes for major pharmacogenes from clinical WES and WGS data with >99% accuracy. These findings support the use of Aldy to repurpose clinical NGS data to inform clinical PGx.


Simulated climate change impact on summer dissolved organic carbon release from peat and surface vegetation: implications for drinking water treatment.
Uncertainty regarding changes in dissolved organic carbon (DOC) quantity and quality has created interest in managing peatlands for their ecosystem services such as drinking water provision. The evidence base for such interventions is, however, sometimes contradictory. We performed a laboratory climate manipulation using a factorial design on two dominant peatland vegetation types (Calluna vulgaris and Sphagnum Spp.) and a peat soil collected from a drinking water catchment in Exmoor National Park, UK. Temperature and rainfall were set to represent baseline and future conditions under the UKCP09 2080s high emissions scenario for July and August. DOC leachate then underwent standard water treatment of coagulation/flocculation before chlorination. C. vulgaris leached more DOC than Sphagnum Spp. (7.17 versus 3.00 mg g(-1)) with higher specific ultraviolet (SUVA) values and a greater sensitivity to climate, leaching more DOC under simulated future conditions. The peat soil leached less DOC (0.37 mg g(-1)) than the vegetation and was less sensitive to climate. Differences in coagulation removal efficiency between the DOC sources appears to be driven by relative solubilisation of protein-like DOC, observed through the fluorescence peak C/T. Post-coagulation only differences between vegetation types were detected for the regulated disinfection by-products (DBPs), suggesting climate change influence at this scale can be removed via coagulation. Our results suggest current biodiversity restoration programmes to encourage Sphagnum Spp. will result in lower DOC concentrations and SUVA values, particularly with warmer and drier summers.