Alkaloids from Stephania venosa as Chemo-Sensitizers in SKOV3 Ovarian Cancer Cells via Akt/NF-κB Signaling.
Crebanine (CN), tetrahydropalmatine (THP), O-methylbulbocapnine (OMBC) and N-methyl tetrahydropalmatine (NMTHP) are isoquinoline derived natural alkaloids isolated from tubers of Stephania venosa. We investigated chemo-sensitizing effects of these alkaloids in ovarian cancer cells and evaluated underlying molecular mechanisms involved in chemo-sensitivity. Detection of cell apoptosis was evaluated by using flow cytometry. Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Chou-Talalay median effect principle was used to evaluate potential drug interactions. Protein analyses were performed on ovarian carcinoma cells using Western blotting upon treatment with anticancer drug and alkaloids. Aporphine alkaloids, such as CN and OMBC, enhanced cisplatin sensitivity in intrinsic cisplatin resistant SKOV3 cells, but not in cisplatin sensitive A2780 cells. Protoberberine alkaloids, such as THP and NMTHP, had no synergistic effect on cisplatin sensitivity in either cell line. Chemo-sensitizing effects of CN and OMBC in SKOV3 cells were mediated via activating apoptosis-induced cell death through caspase-3, -8 and cleaved poly ADP-ribose polymerase (PARP) and via inhibiting anti-apopotic and survival protein expression, such as Bcl-xL, Baculoviral IAP repeat-containing protein 3 (cIAP-2), survivin and interleukin (IL) -6. Cisplatin stimulated protein kinase B (Akt) and nuclear factor-kappaB (NF-κB) signaling pathways, but not mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1) and signal transducer and activator of transcription 3 (STAT3) in SKOV3 cells. Akt/NF-κB signaling was blocked by CN and OMBC leading to increased sensitization to cisplatin. These findings demonstrate that CN and OMBC sensitizes SKOV3 cells to cisplatin via inhibition of Akt/NF-κB signaling and the down regulation of NF-κB mediated gene products. Our results suggest that alkaloids obtained from S. venosa could be used as chemo-sensitizers in ovarian cancer to sensitize and minimize the dose related toxicity of platinum-based chemotherapeutic drugs.


Robotically assisted laparoscopic radical hysterectomy compared with open radical hysterectomy.
Radical hysterectomy is a common and effective treatment of early cervical cancer. Modern advances include the use of robotic assistance to perform equivalent minimally invasive procedures. The purpose was to compare surgical and short-term outcomes, as well as margins, between robotic-assisted laparoscopic radical hysterectomy and open radical hysterectomy. The first 30 cases of robotically assisted type III radical hysterectomy for cervical cancer were compared with the 30 previous cases of open type III radical hysterectomy. Body mass index, length of operation, nodal yield, margins, estimated blood loss, hospital stay, and complications were all documented and compared. The 30 patients undergoing robotically assisted laparoscopic radical hysterectomy were similar in body mass index to the women undergoing open radical hysterectomy (34 kg/m robotic, 32 kg/m open, P = 0.22). The mean operating time was 154 minutes compared with 166 minutes in the open arm (P = 0.36). The mean blood loss was 165 mL compared with 323 mL in the open arm (P = 0.001). The mean pelvic nodal yield was 25 nodes compared with 26 nodes in the open group (P = 0.45). The mean parametrial margin size was not significantly different between groups. The mean postoperative length of stay was 1.4 days compared with 2.8 days for the open cases (P < 0.001). Urinary retention was significantly more common in the robotic arm. Radical surgery for cervical cancer can be accomplished using the da Vinci surgical system (Intuitive Surgical, Sunnyvale, Calif) with acceptable blood loss, operating time, parametrial margins, and nodal yield. Future studies need to address long-term outcomes.


Genetic and chemical evaluation of Trypanosoma brucei oleate desaturase as a candidate drug target.
Trypanosomes can synthesize polyunsaturated fatty acids. Previously, we have shown that they possess stearoyl-CoA desaturase (SCD) and oleate desaturase (OD) to convert stearate (C18) into oleate (C18:1) and linoleate (C18:2), respectively. Here we examine if OD is essential to these parasites. Cultured procyclic (insect-stage) form (PCF) and bloodstream-form (BSF) Trypanosoma brucei cells were treated with 12- and 13-thiastearic acid (12-TS and 13-TS), inhibitors of OD, and the expression of the enzyme was knocked down by RNA interference. The phenotype of these cells was studied. Growth of PCF T. brucei was totally inhibited by 100 µM of 12-TS and 13-TS, with EC(50) values of 40±2 and 30±2 µM, respectively. The BSF was more sensitive, with EC(50) values of 7±3 and 2±1 µM, respectively. This growth phenotype was due to the inhibitory effect of thiastearates on OD and, to a lesser extent, on SCD. The enzyme inhibition caused a drop in total unsaturated fatty-acid level of the cells, with a slight increase in oleate but a drastic decrease in linoleate level, most probably affecting membrane fluidity. After knocking down OD expression in PCF, the linoleate content was notably reduced, whereas that of oleate drastically increased, maintaining the total unsaturated fatty-acid level unchanged. Interestingly, the growth phenotype of the RNAi-induced cells was similar to that found for thiastearate-treated trypanosomes, with the former cells growing twofold slower than the latter ones, indicating that the linoleate content itself and not only fluidity could be essential for normal membrane functionality. A similar deleterious effect was found after RNAi in BSF, even with a mere 8% reduction of OD activity, indicating that its full activity is essential. As OD is essential for trypanosomes and is not present in mammalian cells, it is a promising target for chemotherapy of African trypanosomiasis.


Hypoxic Cancer-Secreted Exosomal miR-182-5p Promotes Glioblastoma Angiogenesis by Targeting Kruppel-like Factor 2 and 4.
Glioblastoma (GBM) is the most lethal primary brain tumor and has a complex molecular profile. Hypoxia plays a critical role during tumor progression and in the tumor microenvironment (TME). Exosomes released by tumor cells contain informative nucleic acids, proteins, and lipids involved in the interaction between cancer and stromal cells, thus leading to TME remodeling. Accumulating evidence indicates that exosomes play a pivotal role in cell-to-cell communication. However, the mechanism by which hypoxia affects tumor angiogenesis via exosomes derived from tumor cells remains largely unknown. In our study, we found that, compared with the parental cells under normoxic conditions, the GBM cells produced more exosomes, and miR-182-5p was significantly upregulated in the exosomes from GBM cells under hypoxic conditions. Exosomal miR-182-5p directly suppressed its targets Kruppel-like factor 2 and 4, leading to the accumulation of VEGFR, thus promoting tumor angiogenesis. Furthermore, exosome-mediated miR-182-5p also inhibited tight junction-related proteins (such as ZO-1, occludin, and claudin-5), thus enhancing vascular permeability and tumor transendothelial migration. Knockdown of miR-182-5p reduced angiogenesis and tumor proliferation. Interestingly, we found elevated levels circulating miR-182-5p in patient blood serum and cerebrospinal fluid samples, and its expression level was inversely related to the prognosis. IMPLICATIONS: Overall, our data clarify the diagnostic and prognostic value of tumor-derived exosome-mediated miR-182-5p and reveal the distinctive cross-talk between tumor cells and human umbilical vein endothelial cells mediated by tumor-derived exosomes that modulate tumor vasculature.


Memory Alterations and White Matter Hyperintensities in Elderly Patients With Hypertension: The ADELAHYDE-2 Study.
The longitudinal ADELAHYDE-2 study aims to identify the factors associated with cognitive impairment/decline and white matter hyperintensities burden. Longitudinal single-center study comprising two visits separated by approximately 7 years. A total of 131 patients completed the two visits. The primary outcome was global memory composite scale, while the secondary outcome was white matter hyperintensities (WMH/Fazekas scale) load. Global memory at visit 2 (V2) was largely influenced by age, smoking status, glycated hemoglobin, and history of stroke already present at visit 1 (V1). These variables accounted for ∼51% of the memory alterations at V2. WMH at V2 was likely influenced by age, left ventricular hypertrophy, diabetes mellitus, carotid intima-media thickness, and body mass index at V1. These findings accounted for ∼37% of the WMH changes at V2. Increase in pulse wave velocity from V1 to V2 showed a trend for association with memory deterioration (adjusted estimates = 0.06; P = .067), whereas smoking and increase in systolic blood pressure (trend) were associated with an increment in WMH (adjusted estimates = 0.49; P = .047 and adjusted estimates = 0.01; P = .08, respectively). On the other hand, angiotensin-converting enzyme inhibitor/angiotensin receptor blockers and statins (trend) were likely to be protective (adjusted estimates for angiotensin-converting enzyme inhibitor/angiotensin receptor blockers = -0.49; P = .049, and adjusted estimates for statins = -0.46; P = .055). Several readily identifiable factors are associated with memory deterioration and WMH, many of which are potentially modifiable. Interventions aimed to control these risk factors need to be tested prospectively in order to assess their cognitive protective value.


Interpretation of differential deficits: the case of aging and mental arithmetic.
A fundamental issue in research on individual differences is the type of evidence sufficient to justify an inference of selective or distinct deficits in relevant theoretical processes. It is proposed that an important consideration is the extent to which the individual differences in 1 variable are independent of those in another variable. Specifically, the suggestion presented here is that a strong conclusion of selective impairment requires evidence that there is significant group-related variance in 1 variable after the variance in the other relevant variable is controlled. Furthermore, an inference that the groups are equivalent on a particular theoretical process requires evidence that the variable presumed to reflect that process has sufficient unique variance to justify the claim that a distinct process is being assessed. The proposed methods are illustrated with two studies comparing adults of different ages in mental arithmetic tasks.


Catecholaminergic regulation of the hypothalamic-pituitary-adrenocortical activity.
The significance and site of adrenergic receptors involved in the control of the hypothalamic-pituitary-adrenal axis (HPA) activity was assessed indirectly by estimation of serum corticosterone levels 1 h after drug administration to conscious rats. Adrenergic drugs were given intracerebroventricularly (icv) and intraperitoneally (ip), the antagonists 15 min prior to the agonists. Noradrenaline, adrenalin and isoproterenol given by either route increased dose-dependently the serum corticosterone levels. The corticosterone response to icv noradrenaline was almost abolished by icv pretreatment with propranolol, a beta-adrenergic antagonist, and yohimbine, and alpha 2-receptor blocker, and was also considerably reduced by prazosin, an alpha 1-adrenergic antagonist. When given ip, these antagonists did not significantly influence the noradrenaline induced corticosterone response, which suggests a suprapituitary site of action of noradrenaline in stimulation of the HPA. The corticosterone response to icv adrenalin was suppressed by prazosin given by either route. The corticosterone response to ip adrenalin was almost abolished by pretreatment with yohimbine, and also significantly diminished by propranolol given by the same route. The increase in corticosterone secretion, induced by isoproterenol given by either route, was abolished by ip injection of propranolol. These results indicate that noradrenaline stimulates the HPA via alpha- and beta-adrenergic receptors, mainly at the suprapituitary level. Adrenalin increases that activity both via central and pituitary alpha- and beta-adrenoceptors. Isoproterenol activates the HPA by stimulation of pituitary beta-receptors.


Characteristics of Adults Managing Vitamins/Supplements and Prescribed Medications-Who Is Using, Not Using, and Abandoning Use of Pillboxes?: A Descriptive Study.
Evidence suggests pillboxes are effective for improving medication adherence. However, prior descriptive studies about pillbox use are limited to studies of older adults or condition-specific studies. This study describes characteristics of adults with chronic conditions and their use of pillboxes. A survey questionnaire link was posted on a social media recruitment page from August 2016 to April 2017. The sample of 179 people was middle-age (47.7 ± 15.4 years), predominantly white (90.4%), educated (>93% educated beyond high school), female (n = 148; men n = 26), married/partner (58.2%), and working full time (55.9%). Pillboxes were used by 66% (n = 118) of the sample at some point; 22.9% reported pillbox abandonment. Compared with people who never used a pillbox, current pillbox users were older (53.2 ± 14.3 vs 42.0 ± 14.4 years; P < .001) and took more vitamins/supplements (3.9 ± 3.8 vs 2 ± 1.8, P = .002) and prescribed medications (4.2 ± 2.2 vs 2.6 ± 1.9, P < .001). Adherence did not differ between groups; pillbox users were more likely to refill medications before running out compared with those abandoning use, P < .001. Age and number of medications may affect pillbox use. Future research should explore barriers to continued use of pillboxes and uptake in younger populations.


Osteoarthritis: another component of metabolic syndrome?
Osteoarthritis (OA) has become a major public health problem not only because of its increasing prevalence worldwide but also because of its frequent association with cardiovascular disease, the leading cause of death in industrialized countries. There is growing evidence that OA is not simply a disease related to aging or mechanical stress of joints but rather a "metabolic disorder" in which various interrelated lipid, metabolic, and humoral mediators contribute to the initiation and progression of the disease process. Indeed, OA has been linked not only to obesity but also to other cardiovascular risk factors, namely, diabetes, dyslipidemia, hypertension, and insulin resistance.


Comparison of LanthaScreen Eu kinase binding assay and surface plasmon resonance method in elucidating the binding kinetics of focal adhesion kinase inhibitors.
An understanding of the dynamics of drug-target interactions is important in the drug discovery process. Information related to the binding kinetics of a drug toward its target or off-target aids in determining the efficacy or toxicity of a drug. Biophysical techniques such as surface plasmon resonance (SPR) have been available for over 20 years, but have been predominantly utilized to characterize protein-protein interactions. With improvements in instrument sensitivity and data analysis software, interactions between proteins (such as kinases) and small molecules have been successfully evaluated. More recently, the LanthaScreen Eu kinase binding assay for characterizing kinase inhibitors has been described. This assay monitors displacement of an Alexa Fluor 647-labeled tracer from the ATP-binding site of an epitope-tagged kinase by a test compound. Such behavior results in a decrease in time-resolved fluorescence energy transfer signal. In this report, a side-by-side comparison of the LanthaScreen Eu kinase binding assay and the SPR method was performed using inhibitors of focal adhesion kinase. The two methods yielded comparable results and identified compounds with time-dependent inhibition and relatively slow dissociation.


Altered cortisol awakening response in posttraumatic stress disorder.
An altered function of the hypothalamic-pituitary-adrenal axis is assumed to be characteristic for Posttraumatic Stress Disorder (PTSD), although there is inconsistent empirical evidence. Only few studies examined the awakening cortisol response and a daytime profile in PTSD. Salivary cortisol levels were measured at seven intervals from awakening until 8 PM in trauma-exposed subjects with (N=29) and without PTSD (N=19) and in 15 non-exposed controls. While the three groups did not differ with respect to their first cortisol level immediately after awakening, the expected cortisol increase to awakening 15-60 min later was significantly lower in PTSD patients compared to non-PTSD subjects and healthy controls. This effect remained stable when trauma-exposed subjects with comorbid major depression were excluded from the analysis. A significant negative correlation between the overall cortisol secretion (AUC(G)) and overall PTSD symptomatology and hyper-arousal symptoms was found. The findings are discussed in light of the hypothesis of a counterregulation of hyper-arousal symptoms and chronic stress in PTSD.


Grafting the alar rim: application as anatomical graft.
Alar rim contour and alar rim grafts have become essential components of rhinoplasty. Ideally, grafts of the nose should be anatomical in shape. So doing might make grafts of the alar rim more robust. The authors considered doing that by applying the graft as a continuous extension of the lateral crus. Twelve patients (two men and 10 women) constituted the study group (seven primary and five secondary cases). Of those, there were five concave rims, two concave rims with rim retraction, two boxy tips, and three cephalically oriented lateral crura. Surgical technique included the following: (1) an open approach was used; (2) a marginal incision that ignored the caudal margin of the lateral crus (the incision went straight posteriorly to a point 5 to 6 mm from the rim margin) was used; (3) a triangular graft was made to cover the exposed vestibular skin; (4) it was secured end to end to the caudal border of the lateral crus; and (5) the poster end was allowed to sit in a small subcutaneous pocket. Follow-up was 11 to 19 months. All 12 patients exhibited good rims as judged by a blinded panel. Rim retraction was not fully corrected in one patient, but no further treatment was required. One patient did require a secondary small rim graft for residual rim concavity. The concept of grafting the alar rim is strongly supported by the authors' results. The modifications the authors applied by designing the graft to be anatomical in shape has been a technical help.


Institutional profile: University of Florida Health Personalized Medicine Program.
The University of Florida (UF) Health Personalized Medicine Program launched in 2012 with CYP2C19 genotyping for clopidogrel response at UF Health Shands Hospital. We have since expanded CYP2C19 genotyping to UF Health Jacksonville and established the infrastructure at UF Health to support clinical implementation for five additional gene-drug pairs: TPMT-thiopurines, IFNL3 (IL28B)-PEG IFN-α-based regimens, CYP2D6-opioids, CYP2D6/CYP2C19-antidepressants and CYP2C19-proton pump inhibitors. We are contributing to the evidence based on outcomes with genotype-guided therapy through pragmatic studies of our clinical implementations. In addition, we have developed a broad array of educational programs for providers, trainees and students that incorporate personal genotype evaluation to enhance participant learning.


First-principles theoretical analysis of silyl radical diffusion on silicon surfaces.
We report results from a detailed analysis of the fundamental radical precursor diffusion processes on silicon surfaces and discuss their implications for the surface smoothness of hydrogenated amorphous silicon (a-Si:H) thin films. The analysis is based on a synergistic combination of first-principles density functional theory (DFT) calculations of SiH(3) radical migration on the hydrogen-terminated Si(001)-(2 x 1) surface with molecular-dynamics (MD) simulations of SiH(3) radical precursor migration on surfaces of a-Si:H films. Our DFT calculations yield activation energies for SiH(3) migration that range from 0.18 to 0.89 eV depending on the local electronic environment on the Si(001)-(2 x 1):H surface. In particular, when no substantial surface relaxation (Si-Si bond breaking or formation) accompanies the hopping of the SiH(3) radical the activation barriers are highest, whereas hopping between nearest-neighbor overcoordinated surface Si atoms results in the lowest radical diffusion barrier of 0.18 eV; this low barrier is consistent with the activation barrier for SiH(3) migration through overcoordinated sites on the a-Si:H surface. Specifically, the analysis of the MD simulations of SiH(3) radical migration on a-Si:H surfaces yields an effective diffusion barrier of 0.16 eV, allowing for the rapid migration of the SiH(3) radical prior to its incorporation in surface valleys; rapid migration and subsequent incorporation constitute the two-step mechanism responsible for the smoothness of plasma deposited a-Si:H thin films.


Microstructural and Mechanical Properties of Alkali Activated Materials from Two Types of Blast Furnace Slags.
This paper investigated the effect of blast furnace slags (BFS) characteristics on the properties achievement after being alkali activated. The physical and chemical characteristics of BFS were determined by X-ray fluorescence (XRF), X-ray Diffraction (XRD) and laser granulometry. Multi-technical characterizations using calorimetry, XRD, Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetry (TG-DTG), scanning electron microscope (SEM), nitrogen sorption and uniaxial compressive strength (UCS) were applied to give an in-depth understanding of the relationship between the reaction products, microstructure and BFS characteristics. The test results show that the microstructure and mechanical properties of alkali activated blast furnace slags (BFS) highly depend on the characteristics of BFS. Although the higher content of basic oxide could accelerate the hydration process and result in higher mechanical properties, a poor thermal stabilization was observed. On the other hand, with a higher content of Fe, the hydration process in alkali activated BFS2 lasts for a longer time, contributing to a delayed compressive strength achievement.


Effect of continuous furosemide infusion on outcome of acute kidney injury.
To evaluate the clinical effects of continuous intravenous infusion with high-dose furosemide on early acute kidney injury (AKI) complicated with acute lung edema. Ninety patients who had been treated by furosemide at routine dose for 12 hour but with unsatisfactory outcomes were selected and subjected to continuous intravenous infusion with high-dose furosemide. The dose was adjusted according to hourly urine output. Serum levels of urea nitrogen, creatinine and potassium, pH, oxygenation index and mechanical ventilation time before and 6, 12, 24, 48 and 72 hour after treatment were compared. The urine outputs before and 6, 12, 24, 48 and 72 hour after treatment were (10.71 ± 1.81), (164.52 ± 21.42), (189.71 ± 29.61), (181.33 ± 23.52), (176.82 ± 24.80) and (164.52 ± 18.91) ml/h respectively. Compared with data before treatment, the serum levels of urea nitrogen, creatinine and potassium significantly decreased while pH and oxygenation index significantly increased after six hour of treatment (P<0.05). After treatment, the kidney functions of 80 patients (88.9%) were completely recovered, without obvious adverse reactions. For patients with early AKI complicated with acute pulmonary edema who cannot be cured by diuretic agent at routine dose, high-dose furosemide increases urine output and improves success rate.


A Simple Cell-based Immunofluorescence Assay to Detect Autoantibody Against the N-Methyl-D-Aspartate (NMDA) Receptor in Blood.
The presence of anti-NMDA receptor autoantibody can cause various neuropsychiatric symptoms in the affected patients, termed anti-NMDA receptor autoimmune encephalitis. Detection of the specific autoantibody against the NMDA receptor in the blood or cerebrospinal fluid (CSF) is essential for the accurate diagnosis of this condition. The NMDA receptor is an ion channel protein complex that contains four subunits, including two mandatory NMDA receptor subunit 1 (NR1) and one or two NMDA receptor subunit 2A (NR2A), NMDA receptor subunit 2B (NR2B), NMDA receptor subunit 2C (NR2C), or NMDA receptor subunit 2D (NR2D). The epitope of anti-NMDA receptor autoantibody was reported to be present at the extracellular N-terminal domain of the NR1 subunit of the NMDA receptor. The goal of this study is to develop a simple cell-based immunofluorescence assay that can be used as a screening test to detect the presence of autoantibodies against NR1 subunit of the NMDA receptor in the blood to facilitate the clinical and basic research of anti-NMDA receptor autoimmune encephalitis.


Towards an accurate representation of electrostatics in classical force fields: efficient implementation of multipolar interactions in biomolecular simulations.
The accurate simulation of biologically active macromolecules faces serious limitations that originate in the treatment of electrostatics in the empirical force fields. The current use of "partial charges" is a significant source of errors, since these vary widely with different conformations. By contrast, the molecular electrostatic potential (MEP) obtained through the use of a distributed multipole moment description, has been shown to converge to the quantum MEP outside the van der Waals surface, when higher order multipoles are used. However, in spite of the considerable improvement to the representation of the electronic cloud, higher order multipoles are not part of current classical biomolecular force fields due to the excessive computational cost. In this paper we present an efficient formalism for the treatment of higher order multipoles in Cartesian tensor formalism. The Ewald "direct sum" is evaluated through a McMurchie-Davidson formalism [L. McMurchie and E. Davidson, J. Comput. Phys. 26, 218 (1978)]. The "reciprocal sum" has been implemented in three different ways: using an Ewald scheme, a particle mesh Ewald (PME) method, and a multigrid-based approach. We find that even though the use of the McMurchie-Davidson formalism considerably reduces the cost of the calculation with respect to the standard matrix implementation of multipole interactions, the calculation in direct space remains expensive. When most of the calculation is moved to reciprocal space via the PME method, the cost of a calculation where all multipolar interactions (up to hexadecapole-hexadecapole) are included is only about 8.5 times more expensive than a regular AMBER 7 [D. A. Pearlman et al., Comput. Phys. Commun. 91, 1 (1995)] implementation with only charge-charge interactions. The multigrid implementation is slower but shows very promising results for parallelization. It provides a natural way to interface with continuous, Gaussian-based electrostatics in the future. It is hoped that this new formalism will facilitate the systematic implementation of higher order multipoles in classical biomolecular force fields.


Oxcarbazepine-induced Stevens-Johnson syndrome: a pediatric case report.
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis are two rare but life-threatening diseases characterized by detachment of epidermis, bullous skin lesions and mucous membrane erosions. Anti-epileptic drugs are highly suspected to be the causative agents. Although carbamazepine (CBZ) is the most associated anti-epileptic drug, oxcarbazepine (OXC), which is a monohydrated derivative of CBZ, is proposed to be safer because of the different metabolism of the two drugs. Herein, we report a case of SJS induced by oxcarbazepine. A 6-year-old boy with benign rolandic epilepsy, admitted to our hospital with generalized maculopapular rash after starting oxcarbazepine. The diagnosis of SJS was made with cytotoxic skin lesions and mucous membrane involvement. After discontinuing of the drug and topical corticosteroid initiation, the lesions were improved. We report this case to attract attention to the serious side effect of this anti-epileptic drug.


Discrepancies Created by Surgeon Self-Reported Operative Time and the Effects on Procedural Relative Value Units and Reimbursement.
To demonstrate discrepancies between operative times in the ACS NSQIP (American College of Surgeons National Surgical Quality Improvement Project) and self-reported operative time from the American Medical Association's Relative Value Scale Update Committee (RUC) and their effect on relative value units (RVU) determination. This is a cross-sectional review of registry data using the ACS NSQIP 2016 Participant User File and the Centers for Medicare & Medicaid Services physician procedure time file for 2018. We analyzed total RVUs for surgeries by operative time to calculate RVU per hour and stratified by specialty. Multivariate regression analysis adjusted for patient comorbidities, age, length of stay, and ACS NSQIP mortality and morbidity probabilities. The surgeon self-reported operative times from the Centers for Medicare & Medicaid Services physician were compared with operative times recorded in the ACS NSQIP, with excess time from RUC estimates termed "overreported time." Analysis of 901,917 surgeries revealed a wide variation in median RVU per hour between specialties. Orthopedics (14.3), neurosurgery (12.9), and general surgery (12.1) had the highest RVU per hour, whereas gynecology (10.2), plastic surgery (9.5), and otolaryngology (9) had the lowest (P<.001 for all comparisons). These results remained unchanged on multivariate regression analysis. General surgery had the highest median overreported operative time (+26 minutes) followed by neurosurgery (+23.5 minutes) and urology (+20 minutes). Overreporting of the operative time strongly correlated to higher RVU per hour (r=0.87, P=.002). Despite reliable electronic records, the AMA-RUC continues to use inaccurate self-reported RUC surveys for operative times. This results in discrepancies in RVU per hour (and subsequent reimbursement) across specialties and a persistent disparity for women-specific procedures in gynecology. Relative value unit levels should be based on the available objective data to eliminate these disparities.


Congenital heart diseases in aged patients: clinical features, diagnosis, and therapeutic indications based on the analysis of a twenty five-year Medline search.
The increasing aging of the population make congenital heart disease more likely to be found in aged patients. No systematic data exist on epidemiology, diagnosis, and therapy in this complex class of patients. The purpose of this study was to provide specific clinical, diagnostic, and therapeutic information concerning patients over 65 with congenital heart disease (CHDs) and to identify the most frequent CHDs in such class of patients. A Medline search was performed to include articles related to congenital heart disease in patients over 65 in worldwide literature from January 1977 to January 2004. Follow-up studies of old, operated CHDs were excluded. The selected papers were divided and numbered on the basis of key words in the title and contents into general, diagnostic, and therapeutic topics. One thousand ninety-five articles met the search criteria. Atrial septal abnormalities, coronary artery anomalies, bicuspid aorta and congenital aortic valve stenosis, and patent ductus arteriosus resulted in the most cited CHDs in the elderly. Clinical manifestations included syncope, supraventricular/ventricular arrhythmias, symptoms of chronic pulmonary hypertension, and congestive heart failure. Echocardiography for the most frequent CHDs and computed tomography and magnetic resonance for complex cases were the main diagnostic tools (277, 104, and 110 citations, respectively). Catheterization plays a marginal role (68 articles). Age is not considered a contraindication to interventions; surgical and endovascular therapy appeared to be similarly useful in managing the most frequent CHDs (88 and 63 citations, respectively), whereas medical treatments were selected for most complex CHDs (71 citations). Patients with CHDs, especially those with low mortality in childhood, are more frequently reaching old age. New imaging techniques and endovascular treatments seem useful in the challenging management of CHDs in aged patients.


Allele frequency net 2015 update: new features for HLA epitopes, KIR and disease and HLA adverse drug reaction associations.
It has been 12 years since the Allele Frequency Net Database (AFND; http://www.allelefrequencies.net) was first launched, providing the scientific community with an online repository for the storage of immune gene frequencies in different populations across the world. There have been a significant number of improvements from the first version, making AFND a primary resource for many clinical and scientific areas including histocompatibility, immunogenetics, pharmacogenetics and anthropology studies, among many others. The most widely used part of AFND stores population frequency data (alleles, genes or haplotypes) related to human leukocyte antigens (HLA), killer-cell immunoglobulin-like receptors (KIR), major histocompatibility complex class I chain-related genes (MIC) and a number of cytokine gene polymorphisms. AFND now contains >1400 populations from more than 10 million healthy individuals. Here, we report how the main features of AFND have been updated to include a new section on 'HLA epitope' frequencies in populations, a new section capturing the results of studies identifying HLA associations with adverse drug reactions (ADRs) and one for the examination of infectious and autoimmune diseases associated with KIR polymorphisms-thus extending AFND to serve a new user base in these growing areas of research. New criteria on data quality have also been included.


First identification of a recombinant form of hepatitis C virus in Austrian patients by full-genome next generation sequencing.
Hepatitis C virus (HCV) intergenotypic recombinant forms have been reported for various HCV genotypes/subtypes in several countries worldwide. In a recent study, four patients living in Austria had been identified to be possibly infected with a recombinant HCV strain. To clarify results and determine the point of recombination, full-genome next-generation sequencing using the Illumina MiSeq v2 300 cycle kit (Illumina, San Diego, CA, USA) was performed in the present study. Samples of all of the patients contained the recombinant HCV strain 2k/1b. The point of recombination was found to be within the HCV NS2 gene between nucleotide positions 3189-3200 based on H77 numbering. While three of four patients were male and had migration background from Chechnya (n = 2) and Azerbaijan (n = 1), the forth patient was a female born in Austria. Three of the four patients including the female had intravenous drug abuse as a risk factor for HCV transmission. While sequencing techniques are limited to a few specialized laboratories, a genotyping assay that uses both ends of the HCV genome should be employed to identify patients infected with a recombinant HCV strain. The correct identification of recombinant strains also has an impact considering the tailored choice of anti-HCV treatment.


MDA-MB-435 cells are derived from M14 melanoma cells--a loss for breast cancer, but a boon for melanoma research.
The tissue of origin of the cell line MDA-MB-435 has been a matter of debate since analysis of DNA microarray data led Ross et al. (2000, Nat Genet 24(3):227-235) to suggest they might be of melanocyte origin due to their similarity to melanoma cell lines. We have previously shown that MDA-MB-435 cells maintained in multiple laboratories are of common origin to those used by Ross et al. and concluded that MDA-MB-435 cells are not a representative model for breast cancer. We could not determine, however, whether the melanoma-like properties of the MDA-MB-435 cell line are the result of misclassification or due to transdifferention to a melanoma-like phenotype. We used karyotype, comparative genomic hybridization (CGH), and microsatalite polymorphism analyses, combined with bioinformatics analysis of gene expression and single nucleotide polymorphism (SNP) data, to test the hypothesis that the MDA-MB-435 cell line is derived from the melanoma cell line M14. We show that the MDA-MB-435 and M14 cell lines are essentially identical with respect to cytogenetic characteristics as well as gene expression patterns and that the minor differences found can be explained by phenotypic and genotypic clonal drift. All currently available stocks of MDA-MB-435 cells are derived from the M14 melanoma cell line and can no longer be considered a model of breast cancer. These cells are still a valuable system for the study of cancer metastasis and the extensive literature using these cells since 1982 represent a valuable new resource for the melanoma research community.


Use of silenced plants in allelopathy bioassays: a novel approach.
Volatile phytohormones or other chemicals can affect processes in distal plant parts but may also influence neighboring plants, and thereby function allelopathically. While this hypothesis has been widely discussed, rigorous tests are lacking. Transgenic plants, silenced in the production of an emitted chemical, are ideal tools to test the hypothesis that the release of a chemical can negatively influence the growth of neighbors (allelopathy). We used isogenic wild type (WT) and genetically transformed plants that lacked the ability to produce ethylene (ir-aco), as both "emitters" and "receivers" of this volatile phytohormone in experiments where receiver plants were only exposed to the headspace of WT or ir-aco emitters, in order to evaluate if natural ethylene releases can function allelopathically. Root growth (a proxy of plant fitness) of WT receivers correlated negatively with the number of WT emitters and headspace ethylene concentrations. Reducing ethylene concentrations in the headspace with the ethylene scrubber, KMnO(4), and using ir-aco seedlings as emitters restored root growth of WT receiver seedlings. 1-Aminocyclopropane-1-carboxylic acid (ethylene biosynthesis substrate) supplementation to WT but not ir-aco emitters inhibited root growth of ir-aco, but not WT receivers, suggesting increased sensitivity to exogenous ethylene of ir-aco seedlings. We conclude that plants genetically silenced in the production of a putative allelochemical are useful in determining if the emitted chemical functions allelopathically.


Thyrotoxic Valvulopathy: Case Report and Review of the Literature.
We report a 42-year-old female who was admitted for abdominal pain, and also endorsed dyspnea, fatigue and chronic palpitations. Past medical history included asthma, patent ductus arteriosus repaired in childhood and ill-defined thyroid disease. Physical examination revealed blood pressure of 136/88 mm Hg and heart rate of 149 beats per minute. Cardiovascular exam revealed an irregularly irregular rhythm, and pulmonary exam revealed mild expiratory wheezing. Abdomen was tender. Electrocardiogram revealed atrial fibrillation with rapid ventricular response which responded to intravenous diltiazem. Labs revealed TSH of < 0.1 mU/L and free T4 of 2.82 ng/dL, a positive TSH-receptor and thyroid peroxidase antibodies suggesting Grave's thyrotoxicosis. A transthoracic echocardiogram reported an ejection fraction of 55-60%, with mild to moderate mitral regurgitation (MR) and moderate to severe tricuspid regurgitation (TR) and dilated right heart chambers. Pulmonary artery systolic pressure was 52 mm Hg. Transesophageal echocardiogram revealed a myxomatous tricuspid valve with thickening and malcoaptation of the leaflets and moderate to severe TR, mild to moderate MR with mild thickening of the mitral valve leaflets. Abdominal ultrasound revealed wall thickening of the gall bladder concerning for acute cholecystitis. She underwent laparoscopic cholecystectomy and was discharged in stable condition on methimazole for her thyroid disease, and on oral diltiazem for rate control and anticoagulation for atrial fibrillation. Follow-up visit with her cardiologist few months later documented absence of cardiac symptoms, and no murmurs were reported on physical examination. This case underscores the importance of maintaining a high index of suspicion for hyperthyroidism when faced with significant newly diagnosed pulmonary hypertension and TR, as treatment of the thyroid abnormalities can reverse these cardiac findings.


Subcutaneous mucor zygomycosis with potential life-threatening visceral complication.
A mass in right supraclavicular fossa in a diabetic patient mimicking tuberculosis (TB) adenitis that ultimately proved to be subcutaneous zygomycosis. A high degree of clinical suspicion is needed for diagnosis especially when these lesions occur at typical sites for the more common indolent infections like TB. This case is being presented not only because of its rarity, but to emphasize the role of early diagnosis and appropriate treatment to prevent serious complications due to proximity to major structures. Fluconazole was used despite not being the ideal drug, solely due to cost constraints. Our patient responded well. However, we do emphasize that response to fluconazole is the exception rather than the rule.


Follow suit on 'PJ paralysis'.
I congratulate Nottingham University Hospital for encouraging patients to get dressed in day clothes, and encourage other acute hospitals to follow suit (news, 8 February) .


[Morphologic characterization of the contemporary dysentery caused by Shigella flexneri 2A].
There is a tendency in St. Petersburg since 1990s to an increase of dysentery morbidity and lethality mainly caused by Flexner Shigella 2a. Morphology of the disease in adults is given on the material of 286 necropsies in 1993-1994 in the S. P. Botkin Infectious Hospital. Males (mean age 50 years) and among them chronic alcoholics prevailed; females (mean age 70 years) were frequently with a grave cardiovascular pathology. Advanced inflammation (fibrinous, ulcerative and hemorrhagic) of all colon and distal part of the small bowel was a common finding. Hemorrhagic infarctions of the ileum which were not described so far were observed in 10% of the cases.


Galectin-3 and S100A9: Novel Diabetogenic Factors Mediating Pancreatic Cancer-Associated Diabetes.
Pancreatic cancer-associated diabetes (PCDM) is a paraneoplastic phenomenon accounting for 1% of new-onset diabetes. We aimed to identify the mediators of PCDM and evaluate their usefulness in distinguishing PCDM from type 2 diabetes. Secreted proteins of MIA PaCa-2 cells were identified by proteomics, and those with ≥10-fold overexpression in transcriptome analysis were assessed by bioinformatics and glucose uptake assay to identify candidate factors. Expression of factors was compared between tumors with and without PCDM by immunohistochemistry. Serum levels were measured in a training set including PC with and without PCDM, type 2 diabetes, pancreatitis, other pancreatic/peripancreatic tumors, and control subjects (n = 50 each). Cutoff values for differentiation between PCDM and type 2 diabetes from the training set were validated in a test set (n = 41 each). Galectin-3 and S100A9 were overexpressed in tumors with PCDM and dose-dependently suppressed insulin-stimulated glucose uptake in C2C12 myotubes. In the training set, serum galectin-3 and S100A9 levels were exclusively increased in patients with PCDM and distinguished PCDM from type 2 diabetes (area under the curve [AUC] galectin-3: 0.73 [95% CI 0.64-0.83]; S100A9: 0.79 [95% CI 0.70-0.87]). Similar results were observed in the test set (AUC galectin-3: 0.83 [95% CI 0.74-0.92]; S100A9: 0.77 [95% CI 0.67-0.87]), with sensitivity and specificity 72.1% and 86.1%, respectively, for galectin-3 and 69.8% and 58.1% for S100A9 in differentiating between PCDM and type 2 diabetes. Galectin-3 and S100A9 are overexpressed in PCDM tumors and mediate insulin resistance. Galectin-3 and S100A9 distinguish PCDM from type 2 diabetes in subjects with new-onset diabetes.


Perceptions of quality health care among parents of children with bleeding disorders.
This study examined how parents of children with bleeding disorders defined quality health care, their expectations for care at the clinic, and indicators of quality health care important to them. Parents (N = 54) answered two open-ended questions and completed the Quality Health Care Questionnaire, which examined the importance of 33 indicators of quality care. The most important indicators of quality care to parents were being included in decisions about their child's care (M = 4.98), being cared for by nurses who are competent and up-to-date (M = 4.94), and being cared for by doctors who are competent and up-to-date (M = 4.94). The results of this study are consistent with earlier research involving other groups of consumers. Parents view quality care as being included in decisions about their child's care and having competent and caring providers with whom they can communicate. In contrast to earlier research, however, waiting time was the least important indicator of quality care.


The effects of fat and protein on glycemic responses in nondiabetic humans vary with waist circumference, fasting plasma insulin, and dietary fiber intake.
The effects of protein and fat on glycemic responses have not been studied systematically. Therefore, our aim was to determine the dose-response effects of protein and fat on the glycemic response elicited by 50 g glucose in humans and whether subjects' fasting plasma insulin (FPI) and diet influenced the results. Nondiabetic humans, 10 with FPI < [corrected] or =40 pmol/L and 10 with FPI >40 pmol/L, were studied on 18 occasions after 10 14-h overnight fasts. Subjects consumed 50 g glucose dissolved in 250 mL water plus 0, 5, 10, or 30 g fat and/or 0, 5, 10, or 30 g protein. Each level of fat was tested with each level of protein. Dietary intake was measured using a 3-d food record. Gram per gram, protein reduced glucose responses approximately 2 times more than fat (P < 0.001) with no significant fat x protein interaction (P = 0.051). The effect of protein on glycemic responses was related to waist circumference (WC) (r = -0.56, P = 0.011) and intake of dietary fiber (r = -0.60, P = 0.005) but was unrelated to FPI or other nutrient intakes. The effect of fat on glycemic responses was related to FPI (r = 0.49, P = 0.029) but was unrelated to WC or diet. We conclude that, across the range of 0-30 g, protein and fat reduced glycemic responses independently from each other in a linear, dose-dependent fashion, with protein having approximately 3-times the effect of fat. A large protein effect was associated with high WC and high dietary-fiber intake, whereas a large fat effect was associated with low FPI. These conclusions may not apply to solid meals. Further studies are needed to determine the mechanisms for these effects.


Percutaneous coronary interventions in facilities without cardiac surgery on site.
Prior to the widespread adoption of intracoronary stent implantation, potential complications of percutaneous coronary intervention (PCI) necessitated the presence of backup cardiac surgery. However, as stent implantation has become the predominant form of PCI, the incidence of emergent cardiac surgery has declined exponentially. Despite this, current guidelines recommend against the performance of elective PCI at hospitals without on-site cardiac surgery and recommend that primary PCI for ST-segment elevation myocardial infarction (STEMI) might be considered at hospitals without backup cardiac surgery. These recommendations are based predominantly on two principles: (1) hospital volume for PCI is strongly associated with clinical outcomes, and (2) results from a large registry study, in which the authors reported a substantial increase in mortality among patients undergoing non-primary/rescue PCI at hospitals without backup cardiac surgery. Since that time, evidence from multiple studies has suggested that performance of PCI at hospitals without backup cardiac surgery is feasible, safe, and both clinically and cost effective. Among STEMI patients, in particular, performance of primary PCI at hospitals without on-site cardiac surgery reduces time to reperfusion and subsequent adverse cardiovascular events as well as likely reducing infarct size. In this review, we will examine the evidence surrounding the performance of PCI for stable and unstable coronary disease at hospitals without on-site backup cardiac surgery.


Suppression of a demand pacemaker in the presence of redundant transvenous right ventricular leads.
A ventricular inhibited demand cardiac pacemaker was inhibited in the presence of intimate contact between the active temporary bipolar electrode ring and a retained inactive permanent bipolar electrode tip. Electromagnetic interference effects, lack of insulation, lead breaks, and loose connections were all ruled out as the cause of pacemaker suppression.


Epicutaneous patch test results in patients with allergic contact dermatitis in Karlovac county--a retrospective survey.
During the 1994-2003 study period, patch testing was carried out in 1102 subjects (740 female and 362 male, mean age 39.8, range 7-81 years) suspected to have allergic contact dermatitis (ACD). Epicutaneous patch test with a standard series of contact allergens was made by the Institute of Immunology, Zagreb, Croatia, according to the Internationational Contact Dermatitis Research Group (ICDRG) rules. During the study period, 399 (36.2%) subjects with one or more positive tests were recorded, and 640 positive tests were observed. The most frequent allergens were nickel sulfate identified in 214 (33.4%), cobalt chloride in 64 (10%), fragrance mix in 60 (9.4%), wood tar in 47 (7.3%), potassium dichromate in 39 (6.1%), balsam of Peru in 29 (4.6%) and other allergens in 187 (29.2%) cases. According to the localization of ACD, hands as the most common site were involved in 535 (48.5%) cases, followed by the face in 167 (15.2%) and other locations in 400 (36.3%) cases. It is concluded that nickel is the principal allergen as the cause of ACD, mostly affecting women. The areas most frequently involved by ACD were the hands and face.


A new approach to maintenance of regenerated autogenous bone volume: delayed relining with xenograft and resorbable membrane.
The aim of this case series is to propose an approach to help maintain autogenous bone grafts. This is done by applying a collagen membrane (CM) and anorganic bovine bone (ABBM) at the time of implant surgery, rather than at the time of ridge augmentation, to avoid volume loss after implants are inserted. Ten patients with severe horizontal bone atrophy were consecutively enrolled in this study. A staged approach was chosen for implant placement following horizontal ridge augmentation. A block graft was harvested from the retromolar area and secured to the recipient site with fixation screws; contour overbuilding was avoided. The width of the ridge was measured before and after horizontal augmentation. After 4 months of healing, implants were inserted, the augmented site was relined with ABBM, and CM was applied to prevent bone volume loss. Another 4 months later, at the time of abutment placement, cone beam computed tomography was performed to quantify the end result. The mean horizontal ridge width prior to treatment was 2.1 ± 0.5 mm. Mean postsurgical crest width was 6.9 ± 0.5 mm. After the 4-month healing period, the mean alveolar crest width was 6.6 ± 0.6 mm. At the time of abutment connection, the mean width of the regenerated ridge, as measured on three-dimensional cone beam images, was 7.7 ± 0.8 mm. Minimal bone loss occurred in unprotected autogenous bone grafts with respect to alveolar bone contour (0.25 ± 0.29 mm). After the implants were inserted, no further remodeling/resorption occurred with sites treated by ABBM and CM relining; moreover, an additional increase in alveolar crest width was evident. The nonresorbable ABBM osseointegrated clinically and radiologically, preventing bone loss prior to implant loading. This layer appeared to maintain the regenerated crest volume.


[Expression of the pectate lyase gene from Erwinia chrysanthemi ENA49 in cells of other Erwinia species].
The gene for a pectate lyase of E. chrysanthemi ENA49 cloned in a recombinant plasmid pPTL1 (a derivative of RSF1010) was transferred into E. carotovora. The pectate lyase determined by the cloned gene was secreted into the cultural medium from the cells of E. crysanthemi EC16. Partial secretion of the enzyme was registered for E. carotovora cells. The major part of EC1 E. chrysanthemi pectate lyase synthesized by E. carotovora cells is accumulated in periplasmic and cytoplasmic fractions. The obtained results suggest the different specificity or efficiency of pectate lyase secretion systems in the studied Erwinia strains.


Methotrexate vs. fumaric acid esters in moderate-to-severe chronic plaque psoriasis: data registry report on the efficacy under daily life conditions.
To compare the clinical efficacy of methotrexate (MTX) vs. fumaric acid esters (FAE) in psoriasis treated under daily life conditions. Data were extracted from a registry (http://www.psoriasisregistry.at) of 272 adult patients with moderate-to-severe chronic plaque psoriasis treated primarily with MTX (n = 72) or FAE (n = 200) between 2004 and 2011. Data from all patients, including those who did not complete at least 3 months of monotherapy, were included in an intention-to-treat (ITT) worst-case analysis. Thirty of 72 (41.7%) patients treated with MTX and 85 of 200 (42.5%) patients treated with FAE discontinued early, mainly due to side-effects or lack of response. Among patients who completed at least 3 months of treatment, the response to primary treatment with MTX vs. FAE did not differ significantly at any time point. In the ITT worst-case analysis at month 3, complete remission rate, PASI90, PASI75 and PASI50 rates were 6%, 7%, 24% and 39% in MTX-treated patients vs. 1%, 5%, 27% and 44% in FAE-treated patients. Overall mean PASI reduction score improved significantly in response to primary MTX and FAE treatment (by 10.6% and 12.6%, respectively) between 3 and 6 months (P = 0.0005; exact Wilcoxon test), but not between 6 and 12 months (P = 0.16). A subset of 32 patients who did not respond satisfactorily to primary treatment with FAE responded better to subsequent MTX therapy (P < 0.0001; paired Wilcoxon test). As shown by retrospective analysis, the primary efficacy of FAE was similar to that of MTX under daily life conditions.


Spontaneous Ventilation Video-Assisted Thoracoscopic Surgery for Geriatric Patients With Non-Small-Cell Lung Cancer.
The aim of the present study was to compare the short-term outcomes between spontaneous ventilation video-assisted thoracic surgery (SV-VATS) and mechanical ventilation video-assisted thoracic surgery (MV-VATS) in the elderly. All patients included in the present study underwent lobectomy, segmentectomy, or wedge resection and lymph node dissection. A retrospective cohor. The first affiliated hospital of Guangzhou Medical University, Guangzhou, China. The present study included 799 elderly patients diagnosed with non-small-cell lung cancer undergoing SV-VATS or MV-VATS. After propensity score matching, 80 patients in the SV-VATS group and 80 patients in the MV-VATS group were analyzed. Patients in the SV-VATS group received spontaneous-ventilation anesthesia, which was administered as follows: intravenous anesthesia + laryngeal mask airway + thoracic paravertebral block + visceral pleural surface anesthesia + thoracic vagus nerve block. Patients in the MV-VATS group received general endotracheal anesthesia. SV-VATS or MV-VATS was performed according to the preference of the patients. There were no significant differences in anesthesia time (226.3 ± 79.8 v 238.5 ± 66.2 min; p = 0.44), surgery time (166.2 ± 102.6 v 170.1 ± 83.4 min; p = 0.66), and number of dissected lymph nodes (5.3 ± 7.5 v 4.4 ± 7.4; p = 0.23) between the two groups. There were significant differences in intraoperative bleeding (61.5 ± 165.1 v 82.2 ± 116.9 mL; p < 0.001). After surgery, the two groups were statistically comparable in terms of hospitalization (17.6 ± 7.6 v 17.2 ± 6.9 days; p = 0.95) and incidence of complications (7.5% v 13.8%; p = 0.20), while there were significant differences in chest tube duration (6.1 ± 3.3 v 4.5 ± 1.2 days; p < 0.001). SV-VATS is feasible and as safe as MV-VATS, and it could be considered as an alternative treatment for the elderly.


A nonadherent cell-based HTS assay for N-type calcium channel using calcium 3 dye.
The N-type calcium channel is a member of the voltage-sensitive calcium channel family and plays a major role in the regulation of neurotransmitter release in the central and peripheral nervous systems. Inhibition of the N-type calcium channel by intrathecal administration of the channel-specific blocker omega-conotoxin MVIIA (ziconotide) is efficacious in the treatment of severe chronic pain. While no orally active small molecules that block the N-type calcium channel are currently available, the discovery of such potentially valuable therapeutics would benefit from a reliable, high throughput assay. However, the assay of N-type calcium channel activity by measuring calcium influx using nonadherent cells in a high throughput fashion has not been achieved before, likely owing to a number of technical hurdles. For example, the measurement of calcium levels in nonadherent cells using conventional calcium indicators, such as Fluo-3 or Fluo-4, requires dyeloading the cells in suspension and subsequent removal of extracellular dye. This limits plate throughput and requires constant handling of the cells. To assay the N-type calcium channel activity using a nonadherent cell line in a high throughput manner, we investigated the application of no-wash calcium assay kits from Molecular Devices Corp. (Sunnyvale, CA): FLIPR Calcium, FLIPR Calcium Plus, and FLIPR Calcium 3. We show here that the FLIPR Calcium 3 assay kit can be used with nonadherent IMR-32 cells to measure potassium-evoked, omega-conotoxin MVIIA-reversible calcium flux with high throughput (15,000 data points/day), high quality (Z approximately 0.6), and minimal handling of the cells. Thus, this assay can be used to reliably and efficiently screen large compound libraries in the search for small molecule N-type calcium channel blockers.


A proposed scenario to improve the Ncut algorithm in segmentation.
In image segmentation, there are many methods to accomplish the result of segmenting an image into k clusters. However, the number of clusters k is always defined before running the process. It is defined by some observation or knowledge based on the application. In this paper, we propose a new scenario in order to define the value k clusters automatically using histogram information. This scenario is applied to Ncut algorithm and speeds up the running time by using CUDA language to parallel computing in GPU. The Ncut is improved in four steps: determination of number of clusters in segmentation, computing the similarity matrix W, computing the similarity matrix's eigenvalues, and grouping on the Fuzzy C-Means (FCM) clustering algorithm. Some experimental results are shown to prove that our scenario is 20 times faster than the Ncut algorithm while keeping the same accuracy.


A comparison of temazepam and flurazepam in terms of sleep quality and residual changes in activation and performance.
A study was carried out in order to compare the effects of 40 mg of temazepam (Euhypnos) and 30 mg of flurazepam on sleep experience, and the residual effects on mood and performance of normal volunteers. The effects of the doses were compared with the identical placebos. A sleep experience questionnaire, an activation questionnaire and a rapid information processing test were administered in the morning, 10 h after ingestion. Both drugs produced the same subjective experience. However, large significant, residual effects of flurazepam were seen on activation and performance (correct detections and reaction times) while temazepam had little effect on activation and produced only a small significant residual impairment of reaction times.


Translocation of classical PKC and cortical granule exocytosis of human oocyte in germinal vesicle and metaphase II stage.
Protein kinase C (PKC) is as a family of serine/threonine kinases that can be activated by Ca2+, phospholipid and diacylglycerol. PKC plays an important role in oocyte maturation and activation. This study was undertaken to investigate classical PKC (cPKC) in human oocyte maturation and activation. Germinal vesicle (GV) and metaphase II (MII) stage oocytes were collected from healthy women. The expression and distribution of cPKC were investigated by immunoflourescence. MII oocytes were treated with PKC activator or inhibitor and imaged using a laser confocal scanning microscope (LCSM). In GV oocytes, PKCalpha, beta1 and gamma were localized to the germinal vesicles, with a weak expression in ooplasm. In MII oocytes, PKCalpha, beta1 and gamma were distributed evenly in ooplasm. After treatment with PKC activator, phorbol 12-myristate 13-acetate (PMA), cPKC translocated to the periphery of oocyte, and cortical granules (CG) exocytosis was found. When the oocytes were treated with PKC inhibitor, staurosporine, no translocation of cPKC and CG exocytosis were found. PKCalpha, beta1 and gamma exist in human oocytes and activation of these subunits could induce CG exocytosis in MII stage.


Phonological fluency norms for Spanish middle-aged and older adults provided by the SCAND initiative (P, M, & R).
Verbal fluency tests are quick and easy to administer neuropsychological measures and are regularly used in neuropsychological assessment. Additionally, phonological fluency is a widely used paradigm that is sensitive to cognitive impairment. This paper offers normative data of phonological verbal fluency (letters P, M, R) for Spanish middle- and older-aged adults, considering sociodemographic factors, and different measures such as the total number of words, errors (perseveration and intrusions), and 15 sec-segmented scores. A total of 1165 cognitively unimpaired participants aged between 50 and 89 years old, participated in the study. Data for P were obtained for all participants. Letters M and R were also administered to a subsample of participants (852) aged 60 to 89 years. In addition, errors and words produced every 15 seconds were collected in the subsample. To verify the effect of sociodemographic variables, linear regression was used. Adjustments were calculated for variables that explained at least 5% of the variance (R2 ≥ .05). Means and standard deviations by age, scaled scores, and percentiles for all tests across different measures are shown. No determination coefficients equal to or greater than .05 were found for sex or age. The need to establish adjustments for the educational level was only found in some of the measures. The current norms provide clinically useful data to evaluate Spanish-speaking natives from Spain aged from 50 to 89 years. Specific patterns of cognitive impairment can be analyzed using these normative data and may be important in neuropsychological assessment.


Exercise Therapy for Parkinson's Disease: Pedaling Rate Is Related to Changes in Motor Connectivity.
Forced-rate lower-extremity exercise has recently emerged as a potential safe and low-cost therapy for Parkinson's disease (PD). The efficacy is believed to be dependent on pedaling rate, with rates above the subjects' voluntary exercise rates being most beneficial. In this study, we use functional connectivity magnetic resonance imaging (MRI) to further elucidate the mechanism underlying this effect. Twenty-seven PD patients were randomized to complete 8 weeks of forced-rate exercise (FE) or voluntary-rate exercise (VE). Exercise was delivered using a specialized stationary bicycle, which can augment patients' voluntary exercise rates. The FE group received assistance from the cycle. Imaging was conducted at baseline, end of therapy, and after 4 weeks of follow-up. Functional connectivity (FC) was determined via seed-based correlation analysis, using activation-based seeds in the primary motor cortex (M1). The change in FC after exercise was compared using linear correlation with pedaling rate. Results of the correlation analysis showed a strong positive correlation between pedaling rate and change in FC from the most affected M1 to the ipsilateral thalamus. This effect persisted after 4 weeks of follow-up. These results indicate that a plausible mechanism for the therapeutic efficacy of high-rate exercise in PD is that it improves thalamo-cortical connectivity.


Factors affecting participation in physical activities in Saudi children with Down syndrome: mothers' perspectives.
Physical activity provides a wide range of benefits. Several studies conducted in various countries have examined factors influencing participation in physical activities in children with Down syndrome. This study aimed to explore factors affecting participation in physical activities in Saudi children with DS, from their mothers' perspectives. In-depth interviews were conducted with 36 mothers of children with Down syndrome to explore facilitators of and barriers to their children's participation in physical activities. All audio recordings were transcribed, validated, reviewed by the authors independently, and organized into themes. Transcript analysis resulted in four facilitators (mother's support and siblings' involvement, involvement of peers, type of physical activity experience, and child's physical ability, behavioral, and psychological status), and six barriers (conditions associated with Down syndrome, family responsibility, social barriers, environmental constraints, electronic devices, and mother's personal psychological barriers). The results regarding perceived barriers indicated a need to develop and implement programs involving family consultation. Environmental modifications should be made to accommodate the needs of children with Down syndrome. A well-organized partnership should be established and activated between private investors and the Ministries of Education, Social Affairs, and Health, to support the promotion of physical activity. Implications for rehabilitation The efficacy of home- and Internet-based programs for Saudi children with Down syndrome (DS) should be evaluated. Cultural and climactic conditions in Saudi Arabia suggest the need for environmental modifications to help children with DS engage in physical activity. Well-organized partnerships between private investors and the Ministries of Education, Social Affairs, and Health may be required to achieve higher participation of children with DS. Planning and designing of strategies, policies, facilities, and programs are required to increase engagement in physical activity and maximize the benefits of participation of children with DS.


A Hematoma Block in the Wrist for a Displaced Distal Radius Fracture Induces Systemic Neurological Symptoms: A Case Report.
Hematoma blocks are considered a relatively simple, safe, and effective technique in the acute setting to provide sufficient anesthesia and allow for closed reduction of displaced distal radius fractures. Complications associated with the utilization of local anesthetics in this setting are rare. We present the case of a patient who had a hematoma block in the wrist and developed short-term systemic neurologic complications likely secondary to systemic absorption of 20 mL of 1% lidocaine without epinephrine via the cancellous bone channels.


Aspects of the ultraviolet photobiology of some T-even bacteriophages.
Bacteriophage T4 DNA metabolism is largely insulated from that of its host, although some host functions assist in the repair of T4 DNA damage. Environmental factors sometimes affect survival and mutagenesis after ultraviolet (UV) irradiation of T4, and can affect mutagenesis in many organisms. We therefore tested the effect of certain environmental factors and host genetic defects upon spontaneous and UV-induced mutagenesis and survival in T4 and some related T-even phages. Plating at pH 9 enhances UV resistance in T4 by about 14% compared to pH 7. The host cAMP regulatory system affects host survival after UV irradiation but does not affect T4 survival. Thermal rescue, the increasing survival of irradiated T4 with increasing plating temperature, occurs also in phage T6, but only weakly in phages T2 and RB69; this temperature effect is not altered by supplementing infected cells with additional Holliday resolvase (gp49) early in infection. Phage RB69 turns out to have almost 50% greater UV resistance than T4, but has a genome of about the same size; RB69 is UV-mutable but does not produce r mutants, which are easily seen in T2, T4, and T6. Spontaneous mutagenesis in T4 shows no dependence on medium and little dependence on temperature overall, but mutation rates can increase and probably decrease with temperature at specific sites. UV mutagenesis is not affected by incubating irradiated particles under various conditions before plating, in contrast to phage S13.


[ICU project to improve pressure ulcer healing rates].
Pressure ulcers represent a very common complication in elderly and patients receiving bedridden care. Inappropriate management of this condition can lead to delayed healing, serious infections and even mortality. The rate of healing for pressure ulcers in our department was 0% between January and June, 2003. We studied this situation and identified the following problems: (1) improper diagnoses; (2) failure to implement a pressure ulcer care protocol; (3) lack of proper instruments to reduce pressure; and (4) failure to care properly for skin following excretion. Nursing knowledge and practice were not updated with new concepts and methods related to clinical pressure ulcer care. To solve these problems and improve pressure ulcer care effectiveness, we organized a special unit in July of the same year, which proceeded to arrange lectures and promotional campaigns, published a standardized care protocol, designed water cushions, and established proper post-excretion care procedures. According to observed results, our department improved its pressure ulcer healing rate by 41.2% within 6 months. This project improved skin care quality and reduced pressure ulcer complications. We recommend that findings and measures be promoted in clinical practice.


Intrafamilial variable phenotype including corticobasal syndrome in a family with p.P301L mutation in the MAPT gene: first report in South America.
Frontotemporal lobar degeneration is a neuropathological disorder that causes a variety of clinical syndromes including frontotemporal dementia (FTD), progressive supranuclear palsy, and corticobasal syndrome (CBS). FTD associated with parkinsonism occurs frequently as a result of mutations in the C9orf72 gene and also in the genes coding for the protein associated with microtubule tau (MAPT) and progranulin (GRN) on chromosome 17 (FTDP-17). Herein, we report an Argentinean family, of Basque ancestry, with an extensive family history of behavioral variant of FTD. Twenty-one members over 6 generations composed the pedigree. An extensive neurologic and neurocognitive examination was performed on 2 symptomatic individuals and 3 nonsymptomatic individuals. Two different phenotypes were identified among affected members, CBS in the proband and FTD in his brother. DNA was extracted from blood for these 5 individuals and whole-exome sequencing was performed on 3 of them followed by Sanger sequencing of candidate genes on the other 2. In both affected individuals, a missense mutation (p.P301L; rs63751273) in exon 10 of the MAPT gene (chr17q21.3) was identified. Among MAPT mutations, p.P301L is the most frequently associated to different phenotypes: (1) aggressive, symmetrical, and early-onset Parkinsonism; (2) late parkinsonism associated with FTD; and (3) progressive supranuclear palsy but only exceptionally it is reported associated to CBS. This is the first report of the occurrence of the p.P301L-MAPT mutation in South America and supports the marked phenotypic heterogeneity among members of the same family as previously reported.


[The molecular basis for increased rigidity of the erythrocyte membrane].
The red blood cell membrane is built on a bilayer lipid matrix associated with proteins. Proteins are located either within the bilayer lipid matrix (intrinsic proteins) or are attached to the hydrophilic surfaces of the bilayer lipid matrix (extrinsic proteins). Intrinsic proteins are held in position by a cytoskeleton. Modifications in both membrane lipids or proteins may alter the red blood cell membrane deformability. The role of the following factors, alone or in combination, in increasing the membrane rigidity, is discussed and reviewed: cholesterol/phospholipids ratio, polyunsaturated fatty acids content, phosphatidylethanolamine methylation, increased intracellular Ca++, protein phosphorylation, membrane protein cross-linking and membrane lipid peroxidation.


The aphid Myzus persicae (Hemiptera: Aphididae) acquires chloroplast DNA during feeding on host plants.
Aphids (Hemiptera: Aphididae) extract nutrients from host plant phloem via stylets that facilitate salivation and sap uptake. When navigating to the phloem, aphids periodically puncture nonvascular cells and sample cell contents, but rarely cause significant cell damage. As a result, aphids are considered "stealthy" feeders. In contrast, insects that do cause damage, such as chewing herbivores, will take up host cell contents-including DNA-into their guts. Researchers can use molecular barcoding methods to identify recent host use patterns of chewing herbivores. This information is valuable for both pest management and basic ecological studies. Because of their stealthy feeding style, it was assumed that host plant DNA could not be recovered from aphids and other Sternorrhyncha. However, several recent studies document host plant DNA uptake by psyllids, which feed in a similar manner to aphids. Therefore, we hypothesized that aphids may also acquire DNA from host plants. Since aphids puncture and sample cytosol contents from cells, we predicted that aphids would be most likely to acquire DNA from chloroplasts. To test this, we performed host feeding and host transfer experiments with Myzus persicae (Sulzer), then used PCR to recover and sequence a region between the trnT and trnF genes from acquired chloroplast DNA. We found that M. persicae readily acquires chloroplast DNA, even prior to phloem contact, and that fragment sizes sufficient for host plant identification can be recovered. Our work suggests that molecular gut content analysis is a viable tool for studying aphid-host interactions.


Clinical Impact of Irregular Protrusion Angle After Coronary Stenting at Culprit Lesions With ST-Elevation Myocardial Infarction - An Intravascular Optical Coherence Tomography Study.
Background: A recent optical coherence tomography (OCT) registry showed that the presence of irregular protrusion (IP) after coronary stenting was a predictor of worse 1-year cardiovascular events. This study evaluated the clinical impact of OCT-detected IP after coronary stenting at ST-elevation myocardial infarction (STEMI) culprit lesions. Methods and Results: In all, 139 consecutive STEMI patients with OCT-detected IP after stenting were analyzed retrospectively. The maximum IP angles were measured and patients with IP were divided into 2 groups (large IP, maximum IP angle ≥180°; small IP, 0°<angle<180°). The primary endpoints were cardiac death, target vessel myocardial infarction, target lesion revascularization, and stent thrombosis at 1 year after the index percutaneous coronary intervention (PCI). Of STEMI patients with IP, 51.8% had large IP. The incidence of the primary endpoints higher was higher in the large than small IP group (12.5% vs. 1.5%, respectively; P=0.018). The occurrence of plaque rupture was an independent predictor of large IP (odds ratio 4.58; 95% confidential interval 1.86-11.27; P=0.001). Conclusions: Maximum IP angle ≥180° was an independent predictor of clinical events in STEMI patients with IP.


Mapping photoreceptor and postreceptoral labelling patterns using a channel permeable probe (agmatine) during development in the normal and RCS rat retina.
The aim of this study was to determine whether agmatine, a channel permeable probe, can identify photoreceptor dysfunction in the Royal College of Surgeons (RCS) retina at an earlier stage to that shown by apoptosis or anatomical markers, and also characterize the neurochemical development of the inner retina in the normal and degenerating rat. We used isolated retinas at different ages incubated in physiological media containing agmatine. Subsequently, postembedding immunocytochemistry was used to determine the number of labelled photoreceptors and the labelling pattern within postreceptoral neurons. Agmatine labelling patterns revealed a sequential development of retinal neurons beginning at postnatal day (PND) 11/12 with most horizontal cells, a few ganglion and amacrine cells, showing a strong signal. The neurochemical development progressed rapidly, and reflects to a large part the known distribution of glutamate receptors, with inner nuclear labelling being evident by PND14, continuing with the same pattern of labelling in adulthood for the control retina. The RCS retina showed markedly reduced agmatine labelling in the inner retina at PND20. A rapid increase in photoreceptor AGB labelling was evident during the degeneration phase. Multiple samples at PND14 and PND16 confirmed a significant increase of labelled photoreceptors in the RCS retina.


Effect of advanced glycation end products, extracellular matrix metalloproteinase inducer and matrix metalloproteinases on type-I collagen metabolism.
The aim of the study was to examine the association among advanced glycation end products (AGEs), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase (MMPs), and investigate whether AGEs affect type I collagen (COL-I) through EMMPRIN or MMPs. A co-culture system with the osteoblast-like cells (MC3T3E1) and mouse RAW264.7 cells was employed to examine the effects of AGE-bovine serum albumin (BSA) (50 mg/l), EMMPRIN antibody (5 mg/l) and AGE-BSA+EMMPRIN antibody separately on COL-I expression for 24 h. Culture media were analyzed for the content of COL-I by ELISA. The effect of different concentrations of AGE-BSA (0, 50, 100, 200 and 400 mg/l) for 24 h was assessed on COL-I levels. Finally, semiquantitative RT-PCR was used to detect the osteoblast COL-I mRNA expression and MMP-2 and MMP-9's PMAO were also measured in the culture medium. COL-I content in the culture medium decreased significantly following treatment with AGE-BSA (P<0.05). EMMPRIN antibody increased COL-I content (P<0.05). EMMPRIN antibody+AGE-BSA increased COL-I significantly (P<0.05). Different concentrations of AGE-BSA increased COL-I mRNA expression significantly compared with the control group (P<0.05), and were enhanced with increasing AGE-BSA concentration (P<0.05). Also MMP-2 and MMP-9 secretion increased significantly (P<0.05), with the increasing AGE-BSA concentration. In conclusion, an increase in AGE levels in vitro stimulates the secretion of EMMPRIN/MMPs, promotes the degradation of COL-I and reduces bone strength.


Cell-specific effects of insulin receptor substrate-1 deficiency on normal and IGF-I-mediated colon growth.
Insulin-like growth factor I (IGF-I) potently stimulates intestinal growth. Insulin receptor substrate-1 (IRS-1) mediates proliferative and antiapoptotic actions of IGF-I in cell lines, but its in vivo relevance in intestine is not defined. This study tested the hypothesis that there is cell type-specific dependence on IRS-1 as a mediator of IGF-I action. Length, mass, crypt cell proliferation, and apoptosis were measured in small intestine and colon of IRS-1-null mice and wild-type (WT) littermates and in colon of IRS-1-null or WT mice expressing IGF-I transgenes. Expression of IGF-I receptor and signaling intermediates was examined in intestine of WT and IRS-1-null mice, cultured intestinal epithelial cells, and myofibroblasts. Absolute IRS-1 deficiency reduced mucosal mass in jejunum and colon, but effects were more pronounced in colon. Muscularis mass was decreased in both segments. In IGF-I transgenics, IRS-1 deficiency significantly attenuated IGF-I-stimulated growth of colonic mucosa and abolished antiapoptotic but not mitogenic effects of IGF-I transgene on crypt cells. IGF-I-induced muscularis growth was unaffected by IRS-1 deficiency. In intestinal epithelial cells, IRS-1 was expressed at higher levels than IRS-2 and was preferentially activated by IGF-I. In contrast, IGF-I activated both IRS-1 and IRS-2 in intestinal myofibroblasts and IRS-2 activation was upregulated in IRS-1-null myofibroblasts. We conclude that the intestinal epithelium but not the muscularis requires IRS-1 for normal trophic actions of IGF-I and that IRS-1 is required for antiapoptotic but not mitogenic effects of IGF-I in the intestinal crypts in vivo.


Investigation of selected genomic deletions and duplications in a cohort of 338 patients presenting with syndromic obesity by multiplex ligation-dependent probe amplification using synthetic probes.
Certain rare syndromes with developmental delay or intellectual disability caused by genomic copy number variants (CNVs), either deletions or duplications, are associated with higher rates of obesity. Current strategies to diagnose these syndromes typically rely on phenotype-driven investigation. However, the strong phenotypic overlap between syndromic forms of obesity poses challenges to accurate diagnosis, and many different individual cytogenetic and molecular approaches may be required. Multiplex ligation-dependent probe amplification (MLPA) enables the simultaneous analysis of multiple targeted loci in a single test, and serves as an important screening tool for large cohorts of patients in whom deletions and duplications involving specific loci are suspected. Our aim was to design a synthetic probe set for MLPA analysis to investigate in a cohort of 338 patients with syndromic obesity deletions and duplications in genomic regions that can cause this phenotype. We identified 18 patients harboring copy number imbalances; 18 deletions and 5 duplications. The alterations in ten patients were delineated by chromosomal microarrays, and in the remaining cases by additional MLPA probes incorporated into commercial kits. Nine patients showed deletions in regions of known microdeletion syndromes with obesity as a clinical feature: in 2q37 (4 cases), 9q34 (1 case) and 17p11.2 (4 cases). Four patients harbored CNVs in the DiGeorge syndrome locus at 22q11.2. Two other patients had deletions within the 22q11.2 'distal' locus associated with a variable clinical phenotype and obesity in some individuals. The other three patients had a recurrent CNV of one of three susceptibility loci: at 1q21.1 'distal', 16p11.2 'distal', and 16p11.2 'proximal'. Our study demonstrates the utility of an MLPA-based first line screening test to the evaluation of obese patients presenting with syndromic features. The overall detection rate with the synthetic MLPA probe set was about 5.3% (18 out of 338). Our experience leads us to suggest that MLPA could serve as an effective alternative first line screening test to chromosomal microarrays for diagnosis of syndromic obesity, allowing for a number of loci (e.g., 1p36, 2p25, 2q37, 6q16, 9q34, 11p14, 16p11.2, 17p11.2), known to be clinically relevant for this patient population, to be interrogated simultaneously.


Multidetector CT Imaging Biomarkers as Predictors of Prognosis in Shock: Updates and Future Directions.
A severe mismatch between the supply and demand of oxygen is the common sequela of all types of shock, which present a mortality of up to 80%. Various organs play a protective role in shock and contribute to whole-body homeostasis. The ever-increasing number of multidetector CT examinations in severely ill and sometimes unstable patients leads to more frequently encountered findings leading to imminent death, together called "hypovolemic shock complex". Features on CT include dense opacification of the right heart and major systemic veins, venous layering of contrast material and blood, densely opacified parenchyma in the right hepatic lobe, decreased enhancement of the abdominal organ, a dense pulmonary artery, contrast pooling in dependent lungs, and contrast stasis in pulmonary veins. These findings are biomarkers and prognostic indicators of paramount importance which stratify risk and improve patient outcomes. In this review, we illustrate the various CT patterns in shock and review the spectrum and prognostic significance of thoraco-abdominal vascular and visceral alarming signs of impending death with the intention of increasing awareness among radiologists and radiographers to prepare for immediate resuscitation when required.


Influence of Ethyl Caproate on the Size of Lipid Vesicles and Yeast Cells.
Ethyl caproate (EC) is a key flavor component of sake. Recently, in sake brewing, an effort has been underway to increase the content of aromatic components such as EC. However, the function of EC in yeast cells remains poorly understood. Therefore, we investigated the effects of EC using cell-sized lipid vesicles. We found that vesicle size decreases in a concentration-dependent manner when EC is contained in lipid vesicles. Furthermore, yeast experiments showed that a strain producing high quantities of EC in its stationary phase decreased in size during EC production. Given caproic acid's (CA) status as the esterification precursor of EC in yeast, we also compared lipid vesicles containing CA with those containing EC. We found that CA vesicles were smaller than EC vesicles of the same concentration. These results suggest that EC production may function apparently to maintain cell size.


Large-scale plasmonic nanodisk array as a biosensing platform fabricated by transfer nanoprinting.
Surface plasmon resonance based on nanostructures has been a powerful analytical tool in rapid detection and analysis of biomolecules. However, the fabrication of nanostructure sensors, such as electron beam lithography and focused ion beam milling, has inherent defects as manufacturing cost, complex process flow, and small fabrication area. In this paper, using the transfer nanoprinting approach based on an ultrathin anodic aluminum oxide membrane, a centimeter-scale ordered periodic Ag-ZnS bilayer nanodisk on Au film with a low cost and simple process is fabricated. A surface plasmon polariton Bloch mode from nanodisk arrays is experimentally demonstrated at normal incident of light. The plasmonic platform exhibits an ideal refractive index bulk sensitivity of up to 438 nm/RIU. Furthermore, by using a polyelectrolyte bilayer with well-defined thickness, the surface sensitivity of the biosensing platform is also investigated. The large-scale plasmonic bilayer nanoparticle biosensing platform has broad application prospects in development of low-cost and high-performance biosensing chips.


Modeling the Batch Sedimentation of Calcium Carbonate Particles in Laboratory Experiments-A Systematic Approach.
The design of continuous thickeners and clarifiers is commonly based on the solid flux theory. Batch sedimentation experiments conducted with solid concentrations still provide useful information for their application. The construction of models for the velocity of settling allows the estimation of the flux of solids throughout time, which can, in turn, be used to find the area of the units required to achieve a given solid concentration in the clarified stream. This paper addresses the numerical treatment of data obtained from batch sedimentation experiments of calcium carbonate particles. We propose a systematic framework to fit a model that is capable of representing the process features that involve (i) the numerical differentiation of data to generate initial estimates for the instantaneous velocity of settling; (ii) the integration of a differential equation to fit the model for the velocity of settling; and (iii) the assessment of the quality of the fit using common statistical indicators. The model used for demonstration has a theoretical basis combined with an empirical component to account for the effect of the particle concentrations and their state of aggregation. The values of the numerical parameters obtained are related to the characteristic dimensions of the aggregates and their mass-length fractal dimensions.


A highly efficient identification of mutants generated by CRISPR/Cas9 using the non‑functional DsRed assisted selection in Aspergillus oryzae.
The CRISPR/Cas9 system has become a great tool for target gene knock-out in filamentous fungi. It is laborious and time-consuming that identification mutants from a large number of transformants through PCR or enzyme-cut method. Here, we first developed a CRISPR/Cas9 system in Aspergillus oryzae using AMA1-based autonomously replicating plasmid and Cas9 under the control of the Aspergillus nidulans gpdA promoter. By the genome editing technique, we successfully obtained mutations within each target gene in Aspergillus oryzae. Then, we put the protospacer sequence of a target gene and its protospacer adjacent motif (PAM) behind the start codon "ATG" of DsRed, yielding the non‑functional DsRed (nDsRed) reporter gene, and the nDsRed reporter gene could be rescued after successful targeted editing. Moreover, this method was also applied by targeting the kojic acid synthesis gene kojA, and the transformants with DsRed activity were found to harbor targeted mutations in kojA. These results suggest that the nDsRed can be used as a powerful tool to facilitate the identification of mutants generated by CRISPR/Cas9 in Aspergillus oryzae.


Changes in excitability of the cortical projections to the human tibialis anterior after paired associative stimulation.
Paired associative stimulation (PAS) based on Hebb's law of association can induce plastic changes in the intact human. The optimal interstimulus interval (ISI) between the peripheral nerve and transcranial magnetic stimulus is not known for muscles of the lower leg. The aims of this study were to investigate the effect of PAS for a variety of ISIs and to explore the efficacy of PAS when applied during dynamic activation of the target muscle. PAS was applied at 0.2 Hz for 30 min with the tibialis anterior (TA) at rest. The ISI was varied randomly in seven sessions (n = 5). Subsequently, PAS was applied (n = 14, ISI = 55 ms) with the TA relaxed or dorsi-flexing. Finally, an optimized ISI based on the subject somatosensory evoked potential (SEP) latency plus a central processing delay (6 ms) was used (n = 13). Motor-evoked potentials (MEPs) were elicited in the TA before and after the intervention, and the size of the TA MEP was extracted. ISIs of 45, 50, and 55 ms increased and 40 ms decreased TA MEP significantly (P = 0.01). PAS during dorsi-flexion increased TA MEP size by 92% (P = 0.001). PAS delivered at rest resulted in a nonsignificant increase; however, when the ISI was optimized from SEP latency recordings, all subjects showed significant increases (P = 0.002). No changes in MEP size occurred in the antagonist. Results confirm that the excitability of the corticospinal projections to the TA but not the antagonist can be increased after PAS. This is strongly dependent on the individualized ISI and on the activation state of the muscle.


A Kinome-Wide Selective Radiolabeled TrkB/C Inhibitor for in Vitro and in Vivo Neuroimaging: Synthesis, Preclinical Evaluation, and First-in-Human.
The proto-oncogenes NTRK1/2/3 encode the tropomyosin receptor kinases TrkA/B/C which play pivotal roles in neurobiology and cancer. We describe herein the discovery of [11C]-(R)-3 ([11C]-(R)-IPMICF16), a first-in-class positron emission tomography (PET) TrkB/C-targeting radiolabeled kinase inhibitor lead. Relying on extensive human kinome vetting, we show that (R)-3 is the most potent and most selective TrkB/C inhibitor characterized to date. It is demonstrated that [11C]-(R)-3 readily crosses the blood-brain barrier (BBB) in rodents and selectively binds to TrkB/C receptors in vivo, as evidenced by entrectinib blocking studies. Substantial TrkB/C-specific binding in human brain tissue is observed in vitro, with specific reduction in the hippocampus of Alzheimer's disease (AD) versus healthy brains. We additionally provide preliminary translational data regarding the brain disposition of [11C]-(R)-3 in primates including first-in-human assessment. These results illustrate for the first time the use of a kinome-wide selective radioactive chemical probe for endogenous kinase PET neuroimaging in human.


Health literacy in old age: results of a German cross-sectional study.
Health literacy is especially important for older people to maintain or enhance remaining health resources and self-management skills. The aim of the study was to determine the level of health literacy and the association between health literacy, demographic and socio-economic factors in German older adults aged 65 years and above stratified by age group. Health literacy was assessed via computer-assisted personal interviews using HLS-EU-Q47 on a representative sample of the German-speaking population. Descriptive statistics, bivariate analyses and logistic regression modelling stratified by age group were conducted to assess health literacy of 475 respondents aged 65 years and above. Overall, 66.3% of all respondents aged 65 years and above had limited health literacy. Limited health literacy was especially prevalent among respondents above 76 years of age (80.6%). Limited health literacy was associated with financial deprivation (OR: 3.05; 95% CI: 1.99-4.67) and limited functional health literacy (OR: 2.16; 95% CI: 1.29-3.61). Financial deprivation was strongest predictor for limited health literacy in the total sample and stratified by age group. Limited health literacy is a frequent phenomenon in German adults aged 65 years and above. Research on health literacy in old age and the role in health disparities is urgently needed.


Acetazolamide and dexamethasone in the prevention of acute mountain sickness.
We randomly assigned 32 healthy backpackers to receive placebo, acetazolamide (250 mg twice a day), dexamethasone acetate (4 mg four times a day), or both drugs in combination to determine the drug efficacy in preventing acute mountain sickness (AMS) at altitudes of 3,650 to 4,050m (12,000 to 13,300 ft). The incidence of AMS was high but symptoms were generally mild. Combined drug therapy was superior to both placebo and single drug therapy in risk reduction. Using acetazolamide alone was moderately beneficial in preventing the occurrence of AMS, although minor side effects were frequent. The use of dexamethasone alone did not significantly reduce the AMS incidence, and discontinuing its use resulted in symptoms suggestive of adrenal insufficiency. For recreational backpackers, routine drug prophylaxis is not recommended, in view of the mild nature of this illness and the adverse effects of medications. The efficacy of combined acetazolamide-dexamethasone therapy warrants further investigation at higher altitudes, where AMS is more severe, and the dexamethasone should be withdrawn gradually to avoid a possible adrenal crisis.


[Discussion on research thinking of traditional Chinese medicine standardization system based on whole process quality control].
The concept of "Quality by design" indicates that good design for the whole life cycle of pharmaceutical production enables the drug to meet the expected quality requirements. Aiming at the existing problems of the traditional Chinese medicine (TCM) industry, the TCM standardization system was put forward in this paper from the national strategic level, under the guidance by the idea of quality control in international manufacturing industry and with considerations of TCM industry's own characteristics and development status. The connotation of this strategy was to establish five interrelated systems: multi-indicators system based on tri-indicators system, quality standard and specification system of TCM herbal materials and decoction pieces, quality traceability system, data monitoring system based on whole-process quality control, and whole-process quality management system of TCM, and achieve the whole process systematic and scientific study in TCM industry through "top-level design-implement in steps-system integration" workflow. This article analyzed the correlation between the quality standards of all links, established standard operating procedures of each link and whole process, and constructed a high standard overall quality management system for TCM industry chains, in order to provide a demonstration for the establishment of TCM whole-process quality control system and provide systematic reference and basis for standardization strategy in TCM industry.