δ-Catenin, a Wnt/β-catenin modulator, reveals inducible mutagenesis promoting cancer cell survival adaptation and metabolic reprogramming.
Mutations of Wnt/β-catenin signaling pathway has essential roles in development and cancer. Although β-catenin and adenomatous polyposis coli (APC) gene mutations are well established and are known to drive tumorigenesis, discoveries of mutations in other components of the pathway lagged, which hinders the understanding of cancer mechanisms. Here we report that δ-catenin (gene designation: CTNND2), a primarily neural member of the β-catenin superfamily that promotes canonical Wnt/β-catenin/LEF-1-mediated transcription, displays exonic mutations in human prostate cancer and promotes cancer cell survival adaptation and metabolic reprogramming. When overexpressed in cells derived from prostate tumor xenografts, δ-catenin gene invariably gives rise to mutations, leading to sequence disruptions predicting functional alterations. Ectopic δ-catenin gene integrating into host chromosomes is locus nonselective. δ-Catenin mutations promote tumor development in mouse prostate with probasin promoter (ARR2PB)-driven, prostate-specific expression of Myc oncogene, whereas mutant cells empower survival advantage upon overgrowth and glucose deprivation. Reprogramming energy utilization accompanies the downregulation of glucose transporter-1 and poly (ADP-ribose) polymerase cleavage while preserving tumor type 2 pyruvate kinase expression. δ-Catenin mutations increase β-catenin translocation to the nucleus and hypoxia-inducible factor 1α (HIF-1α) expression. Therefore, introducing δ-catenin mutations is an important milestone in prostate cancer metabolic adaptation by modulating β-catenin and HIF-1α signaling under glucose shortage to amplify its tumor-promoting potential.


Changes in cerebellar activation pattern during two successive sequences of saccades.
The changes in the cerebellar activation pattern of two successive fMRI scanning runs were determined for visually guided to-and-fro saccades in 12 healthy volunteers familiar with the study paradigm. Group and single subject-analyses revealed a constant activation of the paramedian cerebellar vermis (uvula, tonsils, tuber, folium/declive), which reflects constant ocular motor activity in both runs. A significant decrease in activation of the cerebellar hemispheres found in the second run is best explained by either a decrease in attention or the effects of motor optimization and learning. The significant, systematic changes of the cerebellar activation pattern in two successive runs were not expected, because the ocular motor task was simple, familiar, and highly automated. These findings indicate that similar effects may bias other cerebellar activation studies, in which sensorimotor tasks are repeated in a single session.


The Effect Of Food On The Pharmacokinetic Properties And Bioequivalence Of Two Formulations Of Levocetirizine Dihydrochloride In Healthy Chinese Volunteers.
The aim of this study is to assess the bioequivalence of a new generic formulation and the branded formulation of levocetirizine dihydrochloride in healthy Chinese volunteers under fasting and fed conditions, and food-intake effect on the pharmacokinetic properties is also evaluated. Volunteers were randomly allocated into two groups to receive a single oral dose of generic formulation and branded formulation under fasting or fed conditions, respectively. Blood samples were collected at designated time points. Plasma concentrations of levocetirizine were determined by UFLC-MS/MS. Safety evaluations were carried out through the study. The main pharmacokinetic parameters of the two formulations of levocetirizine were calculated using non-compartmental analysis incorporated in WinNonlin® 7.0 software. Forty-nine volunteers were enrolled; 46 completed the studies. Under fasting and fed conditions, the 90% confidence intervals for the geometric mean of generic/branded ratios were in the range of 94.75-107.24% and 99.98-114.69% for the maximum observed concentration, and 97.13-102.50% and 98.36-103.98% for the area under the concentration-time curve. As a result of food intake before administration, the reduced rate and extent of absorption of levocetirizine were observed. Both formulations were generally well tolerated, with no serious adverse reactions reported. The two formulations demonstrated essentially identical pharmacokinetic profiles and were all well within the FDA/CFDA bioequivalence standards. Meanwhile, food intake can delay the absorption rate and reduced the bioavailability of levocetirizine in healthy Chinese volunteers.


The SGLT2-inhibitor dapagliflozin improves neutropenia and neutrophil dysfunction in a mouse model of the inherited metabolic disorder GSDIb.
Glycogen Storage Disease type 1b (GSDIb) is a genetic disorder with long term severe complications. Accumulation of the glucose analog 1,5-anhydroglucitol-6-phosphate (1,5AG6P) in neutrophils inhibits the phosphorylation of glucose in these cells, causing neutropenia and neutrophil dysfunctions. This condition leads to serious infections and inflammatory bowel disease (IBD) in GSDIb patients. We show here that dapagliflozin, an inhibitor of the renal sodium-glucose co-transporter-2 (SGLT2), improves neutrophil function in an inducible mouse model of GSDIb by reducing 1,5AG6P accumulation in myeloid cells.


[The antimicrobial effect of selected disinfectants in combination with anionic active detergents].
The bactericidal activity of chloramine and hydrogen peroxide can be increased with only few exceptions through admixture of various anionic detergents in dependence of the concentration of the detergent (used concentrations between 0.008-5.3%). On the other hand at chloride of lime only in a few cases was possible to indicate advantageous effects of combination.


Image quality improvement of single-shot turbo spin-echo magnetic resonance imaging of female pelvis using a convolutional neural network.
We have developed a deep learning-based approach to improve image quality of single-shot turbo spin-echo (SSTSE) images of female pelvis. We aimed to compare the deep learning-based single-shot turbo spin-echo (DL-SSTSE) images of female pelvis with turbo spin-echo (TSE) and conventional SSTSE images in terms of image quality.One hundred five and 21 subjects were used as training and test sets, respectively. We performed 6-fold cross validation. In the training process, low-quality images were generated from TSE images as input. TSE images were used as ground truth images. In the test process, the trained convolutional neural network was applied to SSTSE images. The output images were denoted as DL-SSTSE images. Apart from DL-SSTSE images, classical filtering methods were adopted to SSTSE images. Generated images were denoted as F-SSTSE images. Contrast ratio (CR) of gluteal fat and myometrium and signal-to-noise ratio (SNR) of gluteal fat were measured for all images. Two radiologists graded these images using a 5-point scale and evaluated the image quality with regard to overall image quality, contrast, noise, motion artifact, boundary sharpness of layers in the uterus, and the conspicuity of the ovaries. CRs, SNRs, and image quality scores were compared using the Steel-Dwass multiple comparison tests.CRs and SNRs were significantly higher in DL-SSTSE, F-SSTSE, and TSE images than in SSTSE images. Scores with regard to overall image quality, contrast, noise, and boundary sharpness of layers in the uterus were significantly higher on DL-SSTSE and TSE images than on SSTSE images. There were no significant differences in the CRs, SNRs, and respective scores between DL-SSTSE and TSE images. The score with regard to motion artifacts was significantly higher on DL-SSTSE, F-SSTSE, and SSTSE images than on TSE images. The score with regard to the conspicuity of ovaries was significantly higher on DL-SSTSE images than on F-SSTSE, SSTSE, and TSE images (P < .001).DL-SSTSE images showed higher image quality as compared with SSTSE images. In comparison with conventional TSE images, DL-SSTSE images had acceptable image quality while keeping the advantage of the motion artifact-robustness and acquisition time efficiency in SSTSE imaging.


Rural sex work in Cambodia: work characteristics, risk behaviours, HIV, and syphilis.
To identify prevalence and risks factors for syphilis and HIV among rural female sex workers (FSWs) in Cambodia and to describe differences between rural and urban FSWs. Interviews and sera were collected from 114 FSWs and tested for HIV using the Serodia-HIV test and positives confirmed with the enzyme linked immunosorbent assay. Syphilis was tested for with the rapid plasma reagin with passive particle agglutination test for detection of antibody of Treponema pallidum. Study data were merged with data from a study of urban FSWs from Phnom Penh that applied similar questionnaires and sampling design to compare STI prevalence and behaviours. 42% of rural FSWs were HIV positive; 22% had past or current syphilis. In multivariate models HIV was significantly associated with age > or = 25 (OR = 6.1 95% CI: 1.0 to 36.6), a non-commercial partner in the past year (OR= 0.33, 95% CI: 0.11 to 0.93), and prevalence of past or current syphilis (OR = 2.9, 95% CI: 1.0 to 8.8). There was significantly higher active syphilis (14% v 4%), older mean age (25 v 21), fewer daily clients (2 v 5), lower monthly income (61 US dollars v 174 US dollars), and longer duration of sex work (2.3 years v 1.4 years) among rural than among urban FSWs. These findings reveal a high burden of HIV and syphilis among FSWs in rural Cambodia. As FSWs age and become infected with STI/HIV they may move out of cities into less competitive but less savvy markets; their high mobility may contribute to the expansion of the HIV epidemic into rural Cambodia and lower risk populations.


Videomicroscopic imaging of ureteral peristaltic function in rats during cystometry.
Videomicroscopic imaging of the upper urinary tract was performed in 26 female anesthetized rats during bladder filling and micturition. Recordings were made of the pressure of the renal pelvis through a nephrostomy and visualization of the dynamics of the ureteral bolus. Peristaltic velocity, frequency, bolus length, and direction urine bolus propagation were derived on the basis of image processing using indigo carmine for contrast. In addition, nonstop cystometrograms were performed at an infusion rate of 0.22 ml/min characterizing bladder filling and micturition reflexes. Using this setup the pharmacological response of the upper and lower urinary tract dynamics to intravenous oxybutynin and LY274614 was evaluated and compared to observations made with placebo time controls. The data, obtained from the time controls, indicate that there is a significant time-dependent influence on the upper urinary function caused by the experimental methodology in the frequency of ureteral peristalsis and length of the bolus. Oxybutynin produced a significant increase in the length of the but not in the velocity of the bolus. LY274614 depressed pelvic pressure and ureteral frequency and increased bolus length. It is concluded that videomicroscopic imaging, in association with nonstop cystometry, provides a unique method to investigate the pharmacological effects of centrally and peripherally acting drugs on the upper and lower urinary tract function without mechanical manipulation of the ureter.


Identification of a heat shock protein 90 gene involved in resistance to temperature stress in two wing-morphs of Nilaparvata lugens (Stål).
The brown planthopper, Nilaparvata lugens, is one of the most destructive pests damaging rice in Asia and exhibits wing dimorphism, with brachypters possessing severely reduced wings and macropters bearing fully developed wings. Previous studies have shown that macropters are more heat resistant than brachypters. To understand the molecular mechanism underlying the differential thermotolerance abilities of these two morphs, a full-length Hsp gene, NlHsp90 was cloned from N. lugen. Our results showed that the relative expression levels of NlHsp90 in N. lugens females increased with the rise of temperature. Interestingly, NlHsp90 in macropters females could be induced at lower temperature (32°C) than that in brachypters (34°C), and the NlHsp90 mRNA levels in macropters were significantly higher than those in brachypters from 34 to 40°C. In addition, the maximum expression levels of NlHsp90 were achieved much earlier in macropters, and NlHsp90 mRNA levels in macropters were significantly higher than those in brachypters from 1 to 6h of recovery after temperature stress. Furthermore, knockdown of NlHsp90 by dsRNA injection reduced survival in both morphs with a greater reduction in the macropters relative to that of the brachyters. These results indicated that NlHsp90 plays an important role for thermotolerance in N. lugens, and there is difference on induction between two morphs.


Synthesis and evaluation of antibacterial and antibiofilm activities of pyridin-2-yl hexanoate.
Synthesized Pyridin-2-yl hexanoate was screened for its antibacterial and antibiofilm activities using the broth microdilution method and XTT assay respectively. The obtained results revealed that synthesized molecule inhibited the growth of pathogenic bacteria (MICs value ranged from 80 to 640 μg ml-1). Moreover, it induced a strong antibiofilm effect against Gram positive cocci (BIC50 was 366, 378 and 412 μg ml-1 for Streptococcus mutans ATCC 25175, Streptococcus salivarius ATCC 13419 and Streptococcus oralis ATCC 6249 respectively). Basis on these results, pyridin-2-yl hexanoate may be considered as effective compound with antibacterial and antibiofilm activities.


X-ray crystal structure of the blue oxidase ascorbate oxidase from zucchini. Analysis of the polypeptide fold and a model of the copper sites and ligands.
Two crystal forms of the multi-copper protein ascorbate oxidase from Zucchini have been analysed at 2.5 A (1 A = 0.1 nm) resolution and a model of the polypeptide chain and the copper ions and their ligands has been built. Crystal forms M2 and M1 contain a dimer of 140,000 Mr and a tetramer of 280,000 Mr, respectively, in the asymmetric unit. The crystallographic analysis proceeded by multiple isomorphous replacement in M2 followed by solvent flattening and averaging about the local dyad axis. M1 was solved by Patterson search techniques using the M2 electron density. M1 was fourfold averaged. M1 and M2 were combined and the process of averaging repeated in cycles. An atomic model was built into the resulting electron density map and refinement initiated. The current R values of M2 and M1 are 24.5% and 32.6%, respectively. Excellent stereo chemistry was maintained, with root-mean-square deviations of bond lengths and bond angles from average values of 0.02 A and 3.1 degrees, respectively. Each subunit of about 550 amino acid residues has a globular shape with dimensions of 49 A x 53 A x 65 A. It is built up by three domains arranged sequentially on the polypeptide chain and tightly associated in space. The folding of all three domains is of a similar beta-barrel type. It is distantly related to plastocyanin. Each subunit has four copper atoms bound as mononuclear and trinuclear species. The mononuclear copper has two histidine, a cysteine, and a methionine ligand and represents the type-1 copper. It is located in the third domain. The trinuclear cluster has eight histidine ligands. It may be subdivided into a pair of copper atoms with six histidine ligands arranged trigonal prismatic. The pair probably represents the type-3 copper. The remaining copper has two histidine ligands. Its third site of co-ordination is formed by the pair of copper atoms. The fourth ligand may be OH- represented by a small protrusion of electron density. This copper probably is the type-2 copper. The symmetry of the trinuclear cluster is C2 and the ligands are supplied symmetrically by domains 1 and 3. However, domain 1 does not contain a type-1 copper and lacks the characteristic ligands. The unprecedented trinuclear cluster probably represents the oxygen binding and electron storage site.


Interactions between fluorescent labeled phosphatidyl serine and cations.
Phosphatidyl serine (PS) was reacted with dimethylaminoanphthalenesulfonyl (DNS) chloride to produce a fluorescent phospholipid derivative (DNS--PS). By comparison of the emission maximum of the fluorochrome in solvents of varying dielectric constants and in aqueous suspension it was concluded that the DNS moiety of sonicated DNS--PS micells was within the glycerol region of the lipid. Addition of CaC12 to aqueous suspensions of DNS--PS micells caused a shift in the emission maximum from 524 nm to 490 nm, a two-fold increase in the relative fluorescence and an increase in the polarization of fluorescence and finally aggregation. MgC12 produced similar but smaller changes in the fluorescence parameters. In the presence of calcium the thermal transition of the fatty acid tail region could be monitored as an increase in the fluorescence intensity of the DNS group. Without calcium an almost linear decrease in relative fluorescence was observed. These data suggested that calcium caused the fluorochrome to be shifted from the glycerol region of the lipid structure into the fatty acid tail region. These observations indicate how the conformation of a biological membrane might be altered by the interaction between acidic phospholipids and calcium.


LIFE: A Generalizable Autodidactic Pipeline for 3D OCT-A Vessel Segmentation.
Optical coherence tomography (OCT) is a non-invasive imaging technique widely used for ophthalmology. It can be extended to OCT angiography (OCT-A), which reveals the retinal vasculature with improved contrast. Recent deep learning algorithms produced promising vascular segmentation results; however, 3D retinal vessel segmentation remains difficult due to the lack of manually annotated training data. We propose a learning-based method that is only supervised by a self-synthesized modality named local intensity fusion (LIF). LIF is a capillary-enhanced volume computed directly from the input OCT-A. We then construct the local intensity fusion encoder (LIFE) to map a given OCT-A volume and its LIF counterpart to a shared latent space. The latent space of LIFE has the same dimensions as the input data and it contains features common to both modalities. By binarizing this latent space, we obtain a volumetric vessel segmentation. Our method is evaluated in a human fovea OCT-A and three zebrafish OCT-A volumes with manual labels. It yields a Dice score of 0.7736 on human data and 0.8594 ± 0.0275 on zebrafish data, a dramatic improvement over existing unsupervised algorithms.


Real-time PCR-based analysis of BRAF V600E mutation in low and intermediate grade lymphomas confirms frequent occurrence in hairy cell leukaemia.
Hairy cell leukaemia (HCL) is a rare type of B-cell non-Hodgkin lymphoma (B-NHL), which is not known to be associated with any characteristic recurrent karyotypic abnormality. A recent study that used massively parallel whole exome sequencing identified an activating V600E mutation in BRAF, which appeared specific for HCL. Here, we confirm the specificity of BRAF V600E for HCL among low and intermediate grade B-NHL and describe a real-time polymerase chain reaction method for detecting this mutation in cases with low tumour burden. The V600E mutation does not appear to be associated with microsatellite instability, unlike the case in colorectal cancer. Thus, in conjunction with prior data, our results suggest incorporation of BRAF V600E mutation analysis in the diagnostic workup of HCL cases. Additionally, targeting the Ras-Raf-Mek-Erk-Map kinase pathway should be investigated as a potential therapeutic strategy for patients with this disease.


Melting of DNA in confined geometries.
The stability of DNA molecules during viral or biotechnological encapsulation is a topic of active current research. We studied the thermal stability of double-stranded DNA molecules of different lengths in a confined space. Using a statistical model, we evaluate the melting profile of DNA molecules in two geometries: conical and cylindrical. Our results show that not only the confinement, but also the geometry of the confined space plays a prominent role in the stability and opening of the DNA duplex. We find that for more confined spaces, cylindrical confinement stabilizes the DNA, but for less confined spaces conical geometry stabilizes the DNA overall. We also analyse the interaction between DNA sequence and stability, and the evenness with which strand separation occurs. Cylindrical and conical geometries enable a better controlled tuning of the stability of DNA encapsulation and the efficiency of its eventual release, compared to spherical or quasi-spherical geometries.


Dilute alkali pretreatment of softwood pine: A biorefinery approach.
Dilute alkali pretreatment was performed on softwood pine to maximize ethanol and biogas production via a biorefinery approach. Alkali pretreatments were performed with 0-2% w/v NaOH at 100-180°C for 1-5h. The liquid fraction of the pretreated substrates was subjected to anaerobic digestion. The solid fraction of the pretreatment was used for separate enzymatic hydrolysis and fermentation. High ethanol yields of 76.9‒78.0% were achieved by pretreatment with 2% (w/v) NaOH at 180°C. The highest biogas yield of 244mL/g volatile solid (at 25°C, 1bar) was achieved by the pretreatment with 1% (w/v) NaOH at 180°C. The highest gasoline equivalent (sum of ethanol and methane) of 197L per ton of pinewood and the lowest ethanol manufacturing cost of 0.75€/L was obtained after pretreatment with 1% NaOH at 180°C for 5h. The manufacturing cost of ethanol from untreated wood was 4.12€/L.


Evaluating YouTube as a source of patient information for pediatric tracheostomy care.
To evaluate YouTube's usefulness as a source of information concerning pediatric tracheostomy care. On August 10, 2022, the top 50 YouTube search results for "pediatric tracheostomy care" were displayed. Each video was evaluated by a jury of three otolaryngologists with at least 2 years of professional experience in pediatric otolaryngology using DISCERN, scoring system of Journal of the American Medical Association (JAMA), and the Global Quality Score (GQS). After exclusion criteria 24 videos were evaluated. Fifteen of the evaluated videos were produced by health professionals, and the other nine videos were produced by independent users. The average duration of the videos were 337.5 s, varying between 82 s and 1364 s. The average Discern score of videos produced by health professionals was 38.9 ± 13, compared to 36.6 ± 14 for independent users. The mean JAMA score was 1.04 ± 0.68 for health professionals and 1.11 ± 0.94 for independent users. The GQS score was 2.82 ± 0.73 for health professionals and 3.19 ± 0.84 for independent users. There was no statistically significant difference between the two groups for Discern, JAMA, and GQS scoring. YouTube does not seem to be a good option for parents to get useful information about pediatric tracheostomy care at this time. Health professionals should provide websites with high-quality materials to improve awareness of pediatric tracheostomy care.


Persistent hiccups after interventional pain procedures: a case series and review.
Interventional spine procedures are nonsurgical interventions that are commonly used to treat acute and chronic pain. These procedures generally are considered to be safe, but patients may experience transient and minor complications. Hiccups previously have been reported in the pain management setting as a complication after lumbar and thoracic epidural steroid injections and an intrathecal morphine pump infusion. In this case series of 8 patients, we describe hiccups after various interventional procedures, including cervical and lumbar epidural steroid injections, facet joint injections, and sacroiliac joint injections. A comprehensive literature review of hiccups associated with interventional pain procedures is provided, along with the known pathophysiology, etiologies, and treatment options for hiccups. The objective of this case series presentation and literature review is to highlight the importance of recognizing hiccups as a potentially under-reported adverse reaction in the setting of various interventional spine procedures.


AC Josephson effect between two superfluid time crystals.
Quantum time crystals are systems characterized by spontaneously emerging periodic order in the time domain1. While originally a phase of broken time translation symmetry was a mere speculation2, a wide range of time crystals has been reported3-5. However, the dynamics and interactions between such systems have not been investigated experimentally. Here we study two adjacent quantum time crystals realized by two magnon condensates in superfluid 3He-B. We observe an exchange of magnons between the time crystals leading to opposite-phase oscillations in their populations-a signature of the AC Josephson effect6-while the defining periodic motion remains phase coherent throughout the experiment. Our results demonstrate that time crystals obey the general dynamics of quantum mechanics and offer a basis to further investigate the fundamental properties of these phases, opening pathways for possible applications in developing fields, such as quantum information processing.


A sequential extraction methodology for cardiac extracellular matrix prior to proteomics analysis.
Cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) and is a common complication of various cardiovascular diseases. However, little is known about proteins in the cardiac extracellular space. Proteomics analysis of cardiac ECM can be challenging due to the presence of more abundant intracellular proteins, the low degree of solubility of integral ECM proteins, and the presence of abundant posttranslational modifications. Here we describe an extraction methodology based on tissue decellularization, which allows the biochemical subfractionation of extracellular proteins in cardiac tissue. These relatively low-complexity protein fractions are suitable for analysis by gel-LC-MS/MS and other proteomics techniques.


Fluorescence studies on binding of amphiphilic drugs to isolated lamellar bodies: relevance to phospholipidosis.
Lysosomal phospholipid storage disorder in lung tissue was observed during chronic treatment with amphiphilic amine drugs. The prevailing and widely accepted mechanism of phospholipidosis is that amphiphilic drugs bind to phospholipids and make the phospholipids unsuitable substrates for the action of phospholipases. We investigated hydrophobic and hydrophilic binding of fifteen drugs to the phospholipid storage organelle, lung lamellar bodies, isolated from male Sprague-Dawley rats. Hydrophobic interactions were studied using 1,6-diphenyl-1,3,5-hexatriene as a fluorescent probe and hydrophilic binding was studied using 1-anilino-8-naphthalene sulfonate as a fluorescent probe. The binding parameters were calculated using Scatchard equations. Of the fifteen drugs used, nine drugs bound to the hydrophobic moiety of lamellar bodies. The order of binding capacities was promethazine greater than chloramphenicol greater than amiodarone = desethylamiodarone greater than promazine greater than chlorpromazine greater than trimipramine greater than propranolol greater than imipramine much greater than chlorphentermine, phentermine, chloroquine, chlorimipramine, cyclizine and chlorcyclizine. Two binding affinities were calculated for all the bound drugs. Binding affinities to hydrophilic sites of lamellar bodies were calculated in terms of emission coefficients for 1-anilino-8-naphthalene sulfonate in the presence of drugs. Hydrophilic binding was in the order chlorpromazine greater than chlorimipramine greater than promazine greater than trimipramine greater than imipramine greater than chlorcyclizine greater than propranolol greater than promethazine greater than chlorphentermine greater than cyclizine greater than phentermine greater than chloroquine much greater than chloramphenicol, amiodarone and desethylamiodarone. The binding affinities of chlorinated analogs were stronger to hydrophilic sites when compared to the parent compound. Amiodarone, which is known to induce pulmonary phospholipidosis and its major non-polar metabolite, desethylamiodarone, bound strongly to lamellar bodies. These two drugs also inhibit phospholipases in vitro. The drugs with weak phospholipidosis-inducing capacity and extensive in vivo metabolism, namely, imipramine, chlorpromazine and promazine, also bound strongly to lamellar bodies with hydrophilic as well as hydrophobic interactions. On the other hand, chloroquine, which is known to induce phospholipidosis and to inhibit phospholipases, did not bind to lamellar bodies. Two major conclusions could be drawn from this study: one is that the drug interactions with isolated lamellar bodies could be studied using membrane fluorescence probes, 1,6-diphenyl-1,3,5-hexatriene and 1-anilino-8-naphthalene sulfonate; second is that the amphiphilic drugs bind to lamellar bodies, as reported for phospholipid vesicles, and the binding of drugs to lamellar bodies could be correlated with their phospholipidosis-inducing capacity only if


Assessing the stability-indicating properties of alternative potency assays for inactivated influenza vaccine.
Determination of the potency of a vaccine is critical to ensuring that an appropriate dose is delivered, lot-to-lot consistency is maintained, and that the formulation is stable over the life of the vaccine. The potency of inactivated influenza vaccines is determined routinely by the Single Radial Immunodiffusion (SRID) assay. A number of alternative potency assays have been proposed and have been under evaluation in recent years. The aim of this study was to compare a surface plasmon resonance-based assay and two different enzyme linked immunoassays against the current potency assay, SRID, and against mouse immunogenicity when haemagglutinin antigen of the A(H1N1)pdm09 component of an inactivated influenza vaccine is stressed by elevated temperature, low pH and freezing. This analysis demonstrated that the alternative assays had good correspondence with SRID for samples from most stress conditions and that the immunogenicity in mice corresponded with potency in SRID for all stress samples. Subject to further analysis, the assays have been shown to have the potential to possibly replace, and at least complement, SRID.


Numerical simulation of deposition of drifts and salt from multiple super-large seawater cooling towers.
A three-dimensional CFD simulation model was established to study the characteristics of flow, drifts and salt deposition from 6 super-large seawater cooling towers in a power station. In the model, site meteorological data, design parameters of cooling tower, general layout, environmental characteristics, are considered. The results show that: (1) when the wind direction is parallel to the towers, the streams overlap, reducing deposition of drifts and salt onto the ground. (2) The drifts with particle size greater than 550 μm cannot float out of cooling towers. (3) In normal operation of 6 such cooling towers, the resulting salt deposition will not cause serious damage to plants.


WASH inhibits autophagy through suppression of Beclin 1 ubiquitination.
Autophagy degrades cytoplasmic proteins and organelles to recycle cellular components that are required for cell survival and tissue homeostasis. However, it is not clear how autophagy is regulated in mammalian cells. WASH (Wiskott-Aldrich syndrome protein (WASP) and SCAR homologue) plays an essential role in endosomal sorting through facilitating tubule fission via Arp2/3 activation. Here, we demonstrate a novel function of WASH in modulation of autophagy. We show that WASH deficiency causes early embryonic lethality and extensive autophagy of mouse embryos. WASH inhibits vacuolar protein sorting (Vps)34 kinase activity and autophagy induction. We identified that WASH is a new interactor of Beclin 1. Beclin 1 is ubiquitinated at lysine 437 through lysine 63 linkage in cells undergoing autophagy. Ambra1 is an E3 ligase for lysine 63-linked ubiquitination of Beclin 1 that is required for starvation-induced autophagy. The lysine 437 ubiquitination of Beclin 1 enhances the association with Vps34 to promote Vps34 activity. WASH can suppress Beclin 1 ubiquitination to inactivate Vps34 activity leading to suppression of autophagy.


Glycolytic enzyme activities in malignant cells grown in vitro and in vivo.
Cell proximity has a profound effect on the glycolytic enzyme levels observed in mammalian cells grown in vitro. Hexokinase activity generally increased with the increasing cell density of both untransformed and virus-transformed cultures. In contrast, phosphofructokinase and pyruvate kinase activities were generally elevated only in crowded cultures of transformed cells. A spontaneous experimental mouse tumor cell line (EMT6) exhibited cell crowding-dependent elevations in glycolytic enzyme levels when cultured in vitro. The glycolytic enzyme levels of experimental mouse tumors grown in vivo, on the other hand, were constant and normally less than those observed in cultured EMT6 cells.


Chronic ethanol exposure facilitates facial-evoked MLI-PC synaptic transmission via nitric oxide signaling pathway in vivo in mice.
Ethanol (EtOH) exposure causes alterations of motor coordination, balance, behavior, speech, and certain cognitive functions are considered to be caused partly by impairment of cerebellar circuits function and modulation of synaptic transmission. The cerebellar cortical molecular layer interneuron-Purkinje cell (MLI-PC) synapses are critical for various information integration and transmission, which are sensitive to acute and chronic EtOH exposure. The aim of this study is to investigate the effect of chronic ethanol exposure on the facial stimulation-evoked MLI-PC synaptic transmission in urethane-anesthetized mice, by electrophysiological recording and pharmacological methods. Under current-clamp recording conditions, air-puff stimulation of ipsilateral whisker pad evoked MLI-PC synaptic transmission, which expressed an inhibitory component (P1) followed by a pause of simple spike (SS) firing in cerebellar PCs. Chronic ethanol exposure did not change the latency of the facial stimulation-evoked responses in cerebellar PCs, but induced significant enhancement of the stimulation-evoked MLI-PC synaptic transmission, which expressed increases in amplitude of P1 and pause of SS firing. The amplitude of P1 and pause of SS in ethanol exposure group were significant higher than that in control group. Cerebellar surface application of nitric oxide synthesis (NOS) inhibitor, L-NNA (5 mM) significantly decreased the amplitude of P1 and the pause of SS firing in EtOH exposure group, but did no effect on control group. In contrast, cerebellar surface application of NO donor, SNAP (100 μM) significantly increased the amplitude of P1 and the pause of SS firing in control group, but not in EtOH exposure group. These results indicated that chronic EtOH exposure significantly facilitated the sensory-evoked MLI-PC synaptic transmission via NO signaling pathway in mouse cerebellar cortex.


Autopoietic Influence Hierarchies in Pancreatic β Cells.
β cells are biologically essential for humans and other vertebrates. Because their functionality arises from cell-cell interactions, they are also a model system for collective organization among cells. There are currently two contradictory pictures of this organization: the hub-cell idea pointing at leaders who coordinate the others, and the electrophysiological theory describing all cells as equal. We use new data and computational modeling to reconcile these pictures. We find via a network representation of interacting β cells that leaders emerge naturally (confirming the hub-cell idea), yet all cells can take the hub role following a perturbation (in line with electrophysiology).


The effect of pharmacotherapy on prostate volume, prostate perfusion and prostate-specific antigen (prostate morphometric parameters) in patients with lower urinary tract symptoms and benign prostatic obstruction. A systematic review and meta-analysis.
The clinical effect of pharmacotherapy on prostate morphometric parameters is largely unknown. The sole exception is 5α-reductase inhibitors (5-ARI) that reduce prostate volume and prostate-specific antigen (PSA). This review assesses the effect of pharmacotherapy on prostate parameters effect on prostate parameters, namely total prostate volume (TPV), transitional zone volume (TZV), PSA and prostate perfusion. We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) reporting on morphometric parameters' changes after pharmacotherapy, as primary or secondary outcomes. The study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. RCTs' quality was assessed by the Cochrane tool and the criteria of the Agency for Healthcare Research and Quality. The effect magnitude was expressed as standard mean difference (SMD). The study protocol was published on PROSPERO (CRD42020170172). Sixty-seven RCTs were included in the review and 18 in the meta-analysis. The changes after alpha-blockers are comparable to placebo. Long-term studies reporting significant changes from baseline, result from physiologic growth. Finasteride and dutasteride demonstrated large effect sizes in TPV reduction ([SMD]: -1.15 (95% CI: -1.26 to -1.04, p <0.001, and [SMD]:-0.66 (95% CI: -0.83 to -0.49, p <0.001, respectively), and similar PSA reductions. Dutasteride's effect appears earlier (1st vs 3rd month), the changes reach a maximum at month 12 and are sustained thereafter. Phosphodiesterase-5 (PDE-5) inhibitors have no effect on morphometric parameters. Phytotherapy's effect on TPV is non-significant [SMD]: 0.12 (95% CI: -0.03 to 0.27, p = 0.13). Atorvastatin reduces TPV as compared to placebo (-11.7% vs +2.5%, p <0.01). Co-administration of testosterone with dutasteride spares the prostate from the androgenic stimulation as both TPV and PSA are reduced significantly. The 5-ARIs show large effect size in reducing TPV and PSA. Tamsulosin improves perfusion but no other effect is evident. PDE-5 inhibitors and phytotherapy do not affect morphometric parameters. Atorvastatin reduces TPV and PSA as opposed to testosterone supplementation.


Effects of intravenous nicotine on prepulse inhibition in smokers and non-smokers: relationship with familial smoking.
The reinforcing properties of nicotine may be, in part, derived from its ability to enhance certain forms of cognitive processing. Several animal and human studies have shown that nicotine increases prepulse inhibition (PPI) of the startle reflex. However, it remains unclear whether these effects are related to smoking susceptibility. The current study examined the effects of intravenously delivered nicotine on PPI in smokers and non-smokers, as well as its association with a quantitative index of familial smoking. The sample consisted of 30 non-smokers and 16 smokers, who completed an initial assessment, followed on a separate day by a laboratory assessment of PPI prior to and following each of two intravenous nicotine infusions. Separate doses were used in smoker and non-smoker samples. Analyses indicated that both nicotine infusions acutely enhanced PPI among non-smokers, and this enhancement was positively related to the degree of smoking among first and second-degree relatives. Smokers also displayed PPI enhancement after receiving the first infusion, but this effect was unrelated to familial smoking. These data suggest that the PPI paradigm may have utility as an endophenotype for cognitive processes which contribute to smoking risk.


To what degree is palliative care integrated in guidelines and pathways for adult cancer patients in Europe: a systematic literature review.
Palliative Care (PC) aims to improve the quality of life for patients with cancer and their families and its benefits have been demonstrated by several studies. The objective of this systematic review is to assess the integration of PC in the content of guidelines/pathways of adult cancer patients in Europe. We included studies of adult patients with cancer published from 01/01/1995 and 31/12/2013 in Europe in six languages. We searched nine electronic databases, hand-searched six journals and also performed citation tracking. Studies were ranked using Emanuel's Integrated Palliative Care (IPC) criteria, a tool containing 11 domains to assess PC content in guidelines. Two reviewers screened the results and narrative synthesis has been employed. We identified a total of 28,277 potentially relevant articles from which 637 were eligible for full-text screening. The final review included 60 guidelines and 14 pathways. Eighty percent (80%) of the guidelines/pathways emphasize a holistic approach and 66% focus on PC interventions aimed at reducing suffering. Fifty seven percent (57%) did not discuss referral criteria for PC. Of all studies, five fulfilled at least 10/11 IPC criteria. Differences existed with regard to the referral criteria for bereavement care and the continuous adjustment of goals of care. Overall, most of the identified guidelines/pathways highlighted the importance of the holistic approach of IPC. The studies that were found to fulfil at least 10/11 Emanuel's IPC criteria could serve as benchmarks of IPC.


Safety and Feasibility of Quantitative Multiplexed Cytokine Analysis From Office-Based Vitreous Aspiration.
The goals of this study were to evaluate the safety of office-based vitreous sampling, and determine the utility of these samples with multiplex cytokine analysis. Vitreous samples were collected from office-based needle aspiration and the rate of adverse events during follow-up was reviewed. The vitreous cytokine concentrations in a subset of patients with diabetic macular edema (DME) were analyzed using a 42 plex-cytokine bead array. These results were compared with vitreous cytokine concentrations in proliferative diabetic retinopathy (PDR) and controls (macular hole, epiretinal membrane, symptomatic vitreous floaters) from pars plana vitrectomy. An adequate volume of vitreous fluid (100-200 μL) was obtained in 52 (88%) of 59 office-based sampling attempts. The average length of follow-up was 300 days (range, 42-926 days). There were no complications, including cataract, retinal tear or detachment, and endophthalmitis. Two patients (3%) had posterior vitreous detachments within 3 months. Vitreous cytokine concentrations were measured in 44 patients: 14 controls, 13 with DME, and 17 with PDR. The concentration of ADAM11, CXCL-10, IL-8, and PDGF-A were higher in PDR compared with controls and DME. The concentration of IL-6 was higher in PDR compared with controls, but not compared with DME. Office-based vitreous aspiration is safe and yields high-quality samples for multiplex vitreous cytokine analysis. Significant elevations of vitreous cytokines were found in PDR compared with DME and controls, including the novel finding of elevated ADAM11. As such, office-based aspiration is a safe and effective means to identify vitreous factors associated with vitreoretinal disease.


Is cimetidine associated with neutropenia?
Cimetidine is perceived as sufficiently safe that it is being considered for over-the-counter use. However, because of residual concerns about cimetidine-induced neutropenia, a study was conducted to evaluate this association. A population-based, case-control study was undertaken using a large database with Medicaid data from six states. Cases were defined as patients hospitalized with a discharge diagnosis of neutropenia. Medical records were sought to validate the diagnoses of neutropenia and to characterize the severity of disease. Four controls were randomly selected for each case, matched for age, sex, state, and year of diagnosis. The frequency of exposures to cimetidine in the 30 days prior to hospital admission in the cases was compared to that in the identical time periods in the controls. When investigators simultaneously controlled for multiple, potentially confounding variables by using conditional logistic regression, the odds ratio associated with the use of cimetidine was 1.2 (P = 0.33), with a one-sided upper 95% confidence limit of 2.6 for all patients over 20 years of age, and 0.6 (P = 0.30) with a one-sided upper 95% confidence limit of 3.1 for validated cases over 12 years of age. Neither a dose-response relationship was evident (P = 0.899 and P = 0.716, respectively, when excluding the nonusers, or P = 0.245 and P = 0.215, respectively, when including the nonusers). From these data, one can exclude the occurrence of hospitalization with neutropenia or agranulocytosis due to cimetidine occurring more often than once in every 116,000 patients and once in every 573,000 patients receiving a 6-week course of the drug, respectively. If there is an association between cimetidine and neutropenia, it appears to be very small.


Vacuolar ATPase activity is required for endosomal carrier vesicle formation.
A proton pump, the vacuolar ATPase, is known to generate the acidic lumenal environment of endosomes and lysosomes. We have investigated the role of the vacuolar ATPase in endocytic membrane traffic by combining electron microscopy in vivo with a cell-free assay that reconstitutes endosome fusion in vitro. Our observations show that inactivation of this proton pump with bafilomycin A1 has no significant effects on internalization or recycling back to the plasma membrane. However, early endosomes become highly tubular and endocytosed markers do not appear in late endosomes. Our data strongly suggest that, upon inactivation of the proton pump, the formation of a vesicular intermediate between early and late endosomes, which we term endosomal carrier vesicle, is impaired.


Genomic analysis of SBP gene family in Saccharum spontaneum reveals their association with vegetative and reproductive development.
SQUAMOSA promoter binding proteins (SBPs) genes encode a family of plant-specific transcription factors involved in various growth and development processes, including flower and fruit development, leaf initiation, phase transition, and embryonic development. The SBP gene family has been identified and characterized in many species, but no systematic analysis of the SBP gene family has been carried out in sugarcane. In the present study, a total of 50 sequences for 30 SBP genes were identified by the genome-wide analysis and designated SsSBP1 to SsSBP30 based on their chromosomal distribution. According to the phylogenetic tree, gene structure and motif features, the SsSBP genes were classified into eight groups (I to VIII). By synteny analysis, 27 homologous gene pairs existed in SsSBP genes, and 37 orthologous gene pairs between sugarcane and sorghum were found. Expression analysis in different tissues, including vegetative and reproductive organs, showed differential expression patterns of SsSBP genes, indicating their functional diversity in the various developmental processes. Additionally, 22 SsSBP genes were predicted as the potential targets of miR156. The differential expression pattern of miR156 exhibited a negative correlation of transcription levels between miR156 and the SsSBP gene in different tissues. The sugarcane genome possesses 30 SsSBP genes, and they shared similar gene structures and motif features in their subfamily. Based on the transcriptional and qRT-PCR analysis, most SsSBP genes were found to regulate the leaf initial and female reproductive development. The present study comprehensively and systematically analyzed SBP genes in sugarcane and provided a foundation for further studies on the functional characteristics of SsSBP genes during different development processes.


Network strength and dewaterability of flocculated activated sludge.
Works in literature proposed the use of the rheological properties of wastewater sludges as an index for conditioner assessment and control. We demonstrated in this work, on the contrary, that one could not justify the consistency of the commonly used rheological characteristics of the sewage sludge samples taken from the same site but at different dates. A physically relevant index was proposed instead for describing the total network strength, which was hypothesized to correlate the dewatering efficiency of flocculated sludge. Based on this index, the network of a sludge was demonstrated to be largely destroyed after shearing, while the relaxation in an unbound environment or mild pouring over action could partially reinstall the structure. Moreover, a uniform shear rate field could produce network of greater strength when compared with that conditioned in a stirred tank.


A pressing diagnosis - a compromised testicle secondary to compartment syndrome.
Hydrocoeles are a common cause of scrotal swelling and discomfort in a significant proportion of men. We report a case of compartment syndrome within the tunica vaginalis. This is an unusual and previously unreported complication of a hydrocoele.


Clinical benefit of glycoprotein IIb/IIIa blockade with Abciximab is independent of gender: pooled analysis from EPIC, EPILOG and EPISTENT trials. Evaluation of 7E3 for the Prevention of Ischemic Complications. Evaluation in Percutaneous Transluminal Coronary Angioplasty to Improve Long-Term Outcome with Abciximab GP IIb/IIIa blockade. Evaluation of Platelet IIb/IIIa Inhibitor for Stent.
We sought to determine the efficacy and safety of platelet glycoprotein IIb/IIIa receptor (GP IIb/IIIa) blockade with abciximab in women undergoing percutaneous coronary intervention. Although gender differences in response to platelet glycoprotein IIb/IIIa receptor blockade have been described, there have been no large clinical studies to assess these differences. Outcomes were determined using meta-analysis technique. In the pooled analysis, the primary end point of death, myocardial infarction (MI) or urgent revascularization within 30 days was reduced from 11.3% to 5.8% (p<0.001) in men and from 12.7% to 6.5% (p<0.001) in women treated with abciximab. At six months, death, MI or urgent revascularization was reduced from 14.1% to 8.3% (p<0.001) in men and 16.0% to 9.9% (p<0.001) in women receiving abciximab. At one year, mortality was reduced from 2.7% to 1.9% (p = 0.06) in men and 4.0% to 2.5% (p = 0.03) in women treated with abciximab. Major bleeding events occurred in 2.9% versus 3.0% (p = 0.96) of women and 2.7% versus 1.3% (p = 0.003) of men treated with placebo versus abciximab, respectively. Minor bleeding events occurred in 4.7% versus 6.7% (p = 0.01) of women and 2.3% versus 2.2% (p = 0.94) of men treated with placebo versus abciximab, respectively. This pooled analysis demonstrated no gender difference in protection from major adverse outcomes with GP IIb/IIIa inhibition with abciximab. Although women had higher rates of both major and minor bleeding events with abciximab compared with men, major bleeding in women was similar with and without abciximab. There was a small increased risk of minor bleeding with abciximab in women.


A twisted four-sheeted model for an amyloid fibril.
The formation of amyloid fibers and their deposition in the body is a characteristic of a number of devastating human diseases. Here, we propose a structural model, based on X-ray diffraction data, for the basic structure of an amyloid fibril formed by using the variants of the B1 domain of IgG binding protein G of Streptococcus. The model for the fibril incorporates four beta sheets in a bundle with a diameter of 45 A. Its cross-section, or layer, consists of four strands, one strand from each sheet. Layers stack on top of each other to form the fibril, which has an overall helical twist with a periodicity of about 154 A. Each strand interacts in a parallel fashion with the strands in the layers above and below it, in an infinite beta sheet. Some geometric features of this model and the logic behind it may be applicable for constructing other related cross-beta amyloid fibrils.


Paxillin genes and actomyosin contractility regulate myotome morphogenesis in zebrafish.
Paxillin (Pxn) is a key adapter protein and signaling regulator at sites of cell-extracellular matrix (ECM) adhesion. Here, we investigated the role of Pxn during vertebrate development using the zebrafish embryo as a model system. We have characterized two Pxn genes, pxna and pxnb, in zebrafish that are maternally supplied and expressed in multiple tissues. Gene editing and antisense gene knockdown approaches were used to uncover Pxn functions during zebrafish development. While mutation of either pxna or pxnb alone did not cause gross embryonic phenotypes, double mutants lacking maternally supplied pxna or pxnb displayed defects in cardiovascular, axial, and skeletal muscle development. Transient knockdown of Pxn proteins resulted in similar defects. Irregular myotome shape and ECM composition were observed, suggesting an "inside-out" signaling role for Paxillin genes in the development of myotendinous junctions. Inhibiting non-muscle Myosin-II during somitogenesis altered the subcellular localization of Pxn protein and phenocopied pxn gene loss-of-function. This indicates that Paxillin genes are effectors of actomyosin contractility-driven morphogenesis of trunk musculature in zebrafish. Together, these results reveal new functions for Pxn during muscle development and provide novel genetic models to elucidate Pxn functions.


Sources of individual differences in the speed of naming objects and actions: the contribution of executive control.
We examined the contribution of executive control to individual differences in response time (RT) for naming objects and actions. Following Miyake et al., executive control was assumed to include updating, shifting, and inhibiting abilities, which were assessed using operation span, task-switching, and stop-signal tasks, respectively. Experiment 1 showed that updating ability was significantly correlated with the mean RT of action naming, but not of object naming. This finding was replicated in experiment 2 using a larger stimulus set. Inhibiting ability was significantly correlated with the mean RT of both action and object naming, whereas shifting ability was not correlated with the mean naming RTs. Ex-Gaussian analyses of the RT distributions revealed that updating ability was correlated with the distribution tail of both action and object naming, whereas inhibiting ability was correlated with the leading edge of the distribution for action naming and the tail for object naming. Shifting ability provided no independent contribution. These results indicate that the executive control abilities of updating and inhibiting contribute to the speed of naming objects and actions, although there are differences in the way and extent these abilities are involved.


PARP-1-dependent 3-nitrotyrosine protein modification after DNA damage.
3-nitrotyrosine (NO2-Tyr) is thought to be a specific marker of cell injury during oxidative damage. We have evaluated the role of poly(ADP-ribose)polymerase-1 (PARP-1) in protein nitration after treatment of immortalized fibroblasts parp-1+/+ and parp-1-/- with the alkylating agent 2'-methyl-2'-nitroso-urea (MNU). Both cell lines showed increased iNOS expression following MNU treatment in parallel with a selective induction of tyrosine nitration of different proteins. PARP-1 deficient cells displayed a delayed iNOS accumulation, reduced number of nitrated proteins, and a lower global nitrotyrosine "footprint." We have identified the mitochondrial compartment as the major site of oxidative stress during DNA damage, being MnSOD one of the NO2-Tyr-modified proteins, but not in parp-1-/- cells. These results suggest that NO-derived injury can be modulated by proteins involved in the response to genotoxic damage, such as PARP-1, and may account for the limited oxidative injury in parp-1 knockout mice during carcinogenesis and inflammation.


Surgical treatment of advanced medication-related osteonecrosis of the jaws: Comparison of soft tissue closure techniques and evaluation of side effects.
The aim of the study was to evaluate the surgical outcome of patients with advanced medication-related osteonecrosis of the jaw (MRONJ) treated with different flaps for soft tissue closure. Only MRONJ stage III patients with a minimum follow-up of 6 months were included in our prospective study. The soft tissue closure techniques were: Mylohyoid Muscle Flap, Buccal Fat Flap and mucoperiosteal flap alone. Potential risk factors and surgical side effects were analyzed. Relapses occurred in 12 of 44 included cases and 38 reached mucosal integrity within the follow-up. Cases treated with the muscle or fat flap showed better results regarding the recurrence rate (p < 0.001) and soft tissue healing (p = 0.002): only 3 of 33 developed a relapse, and 31 of 33 reached mucosal integrity. The outcome was worse if MRONJ occurred at the front areas of the jaw (p = 0.025). Postoperative, the pain level was reduced significantly (p < 0.001). Partial hypoesthesia of the lip arose in 18 cases. An impairment of the long-term prosthetic rehabilitation has not been seen.Patients with MRONJ stage III undergoing surgery benefit from extensive soft tissue closure.


Implications of the Cardiac Arrhythmia Suppression Trial for antiarrhythmic drug treatment.
The Cardiac Arrhythmia Suppression Trial (CAST) is a randomized, placebo-controlled, double-blind, multicenter clinical trial involving 27 centers and more than 100 hospitals in North America and Europe to test the 1-tailed hypothesis that suppression of ventricular arrhythmias in patients with left ventricular dysfunction after myocardial infarction will reduce arrhythmic death. Since April 18, 1989, the CAST is enrolling patients aged less than 80 years with greater than or equal to 6 ventricular premature complexes and left ventricular ejection fraction less than or equal to 40%. Sustained ventricular tachycardia, class IV congestive heart failure or class IV angina pectoris are exclusion criteria. During a prerandomization period, antiarrhythmic drugs are titrated to suppress ventricular arrhythmias. If greater than or equal to 80% suppression is achieved during open-label titration, patients are randomized to the effective dose or to a matched placebo. If only partial suppression (1 to 79%) is achieved, patients are eligible for a substudy that randomizes them to the best dose found during open-label titration or to placebo. The only patients not randomized to treatment are those with increased arrhythmias or drug intolerance during titration. On April 18, 1989, encainide and flecainide were removed from the CAST because these drugs increased the death rate 2.5-fold. There were no imbalances in baseline risk variables between the encainide/flecainide group and the placebo group that might explain the adverse treatment effect. There was remarkable uniformity of the adverse effect across all subgroups. There were no subgroups that benefited from treatment; all were either harmed or not evaluable.(ABSTRACT TRUNCATED AT 250 WORDS)


Laparoscopic splenectomy for autoimmune hemolytic anemia in patients with chronic lymphocytic leukemia: a case series and review of the literature.
The purpose of this study was to evaluate the safety and efficacy of laparoscopic splenectomy in patients with chronic lymphocytic leukemia (CLL) complicated by autoimmune hemolytic anemia. A series of nine such patients who underwent this procedure at our institution between August 1997 and September 2001 were retrospectively reviewed. Seven of 9 patients who underwent laparoscopic splenectomy for CLL and autoimmune hemolytic anemia achieved a complete response. One patient who initially responded relapsed 12 weeks postoperatively. Therefore, six of 9 patients showed sustained responses with a mean follow-up of 2 years, consistent with other published series. Two patients had no response, one of whom died within 3 weeks of surgery from transformed Hodgkin's disease. The only other postoperative complication occurred in a patient who developed pneumonia. We conclude that laparoscopic splenectomy is a safe and effective treatment for autoimmune hemolytic anemia in patients with CLL who fail medical therapy.


Effects of procainamide and sotalol on restitution properties, dispersion of refractoriness, and ventricular fibrillation activation patterns in pigs.
Interest in combining antiarrhythmic drugs has been prompted by the lack of efficacy of monotherapies and the toxicity resulting from high doses of individual agents. We tested the hypothesis that procainamide and sotalol combined have greater beneficial effects on restitution, on the dispersion of refractoriness, and on decreasing the complexity of ventricular fibrillation (VF) than either drug alone. Six open-chest pigs received intravenous procainamide (15 mg/kg load and 50 microg/kg/min maintenance) followed by sotalol (1.5 mg/kg). Another six pigs received sotalol first and procainamide second. Before drugs and after each drug, 20-second episodes of electrically induced VF were recorded from a 21 x 24 unipolar electrode plaque (2 mm spacing) sutured on the lateral posterior left ventricular epicardium. Restitution properties and dispersion of refractoriness were estimated from activation recovery intervals during pacing. The combination of the two drugs reduced the maximum slope of the restitution curve and during VF reduced the number of wavefronts, the activation rate, the percentage of wavefront families exhibiting reentry, and the conduction velocity more than either drug alone. In addition, in the group that received sotalol first, both drugs together reduced the SD and the coefficient of variation of the spatial dispersion of refractoriness compared with baseline. Procainamide and sotalol combined have greater beneficial effects on restitution properties, dispersion of refractoriness, and the complexity of VF than either drug alone compared with baseline.


A highly stable and sensitive chemically modified screen-printed electrode for sulfide analysis.
We report here a highly stable and sensitive chemically modified screen-printed carbon electrode (CMSPE) for sulfide analysis. The CMSPE was prepared by first ion-exchanging ferricyanide into a Tosflex anion-exchange polymer and then sealing with a tetraethyl orthosilicate sol-gel layer. The sol-gel overlayer coating was crucial to stabilize the electron mediator (i.e., Fe(CN)6(3-)) from leaching. The strong interaction between the oxy-hydroxy functional group of sol-gel and the hydrophilic sites of Tosflex makes the composite highly rigid to trap the ferricyanide mediator. An obvious electrocatalytic sulfide oxidation current signal at approximately 0.20 V versus Ag/AgCl in pH 7 phosphate buffer solution was observed at the CMSPE. A linear calibration plot over a wide range of 0.1 microM to 1mM with a slope of 5.6 nA/muM was obtained by flow injection analysis. The detection limit (S/N=3) was 8.9 nM (i.e., 25.6 ppt). Practical utility of the system was applied to the determination of sulfide trapped from cigarette smoke and sulfide content in hot spring water.


A pilot study of repeated 5-aminolevulinic acid photodynamic therapy for cervical intraepithelial neoplasia by a newly developed intravaginal irradiation system.
The pilot study was performed to assess the usefulness of a newly developed patientand physician-friendly intravaginal irradiation system for photodynamic therapy with 5-aminolevulinic acid (5-ALA PDT) for cervical intraepithelial neoplasia (CIN). We used an intravaginal balloon applicator to make the cervix upright and to adjust the position and direction of the laser source in the vagina, resulting in minimal patient discomfort and minimal effort required by the physician during irradiation. Ten outpatients of CIN2 or 3 with high-risk human papillomavirus (HPV) infection without a history of HPV vaccination were treated by 5-ALA PDT. Each patient underwent PDT four times every two weeks. Nine patients showed pathological improvement, and the HPV clearance rate was 80%, and no recurrence was observed at two-years follow-up. Serum anti-HPV16 antibodies were detected in seven patients, and the antibody levels of three patients were high, equivalent to those acquired after HPV vaccination. Our newly developed irradiation system enabled easy repeat 5-ALA PDT in the outpatient clinic resulting in the improvement of CIN lesions and HPV clearance. Our results also suggested that repeated 5-ALA PDT might enhance HPV antibody production in CIN patients.


Long-Term Outcomes of Surgical Ablation for Atrial Fibrillation: Impact of Ablation Lesion Sets.
The lesion sets for surgical ablation of atrial fibrillation (AF) that provide optimal outcomes have remained controversial. We evaluated the effects of left-atrial (LA) ablation of AF compared with bi-atrial (BA) ablation on the clinical and rhythm outcomes, and examined the predictors for AF recurrence and permanent pacing in consideration of ablation lesion sets. Between 2001 and 2018, 1,965 patients underwent surgical ablation during cardiac surgery at our institution. Among these, 796 and 1,169 patients underwent LA and BA ablation, respectively. The clinical outcomes were evaluated after propensity score adjustment, with death accounting for a competing event. The probability of AF recurrence was estimated with the generalized estimating equations model. The patients with BA ablation had morbidities greater than those with LA ablation. The probability of AF recurrence at 1 and 5 years was 13.9% and 37.1% in patients with LA ablation, and 11.2% and 30.1% in those with BA ablation (hazard ratio [HR]: 1.24; 95% confidence interval [CI]: 0.96-1.61; P = 0.100). After adjustment, LA ablation was associated with a decreased risk of early death (<30 days) (odds ratio [OR]: 0.56; 95% CI: 0.31-0.96; P = 0.041) and new-onset dialysis (OR 0.47; 95% CI: 0.27-0.78; P = 0.003). However, the risk of overall mortality (HR: 1.03; 95% CI: 0.75-1.41; P = 0.878) and permanent pacing (HR: 0.68; 95% CI: 0.43-1.06; P = 0.091) was comparable between the 2 groups. The risk of AF recurrence and adverse events was comparable between the 2 ablation lesion sets. BA ablation was not related to an increased risk of permanent pacing.


Review of Mixed-Methods Research in Nursing.
Inadequate justification for using mixed-methods and inadequate data integration compromises the rigor of mixed-methods studies, and data integration remains a challenge for nurse researchers. The aim of the study was to determine the 5-year prevalence of mixed-methods research in nursing journals and to determine the extent of integration of qualitative and quantitative findings. Ten journals were hand-searched, and additional search was conducted within three databases. Prevalence was calculated by counting the number of published mixed-methods studies divided by the number of published studies over 5 years. Three reviewers independently performed methodological assessment using a checklist based on guidelines by expert methodologists. Prevalence of mixed-methods studies was 1.89%. Concerning methodological assessment, of 175 studies, 29% did not provide an explicit label of the study design and four studies incorrectly labeled the design. In total, 31% of the studies did not justify using mixed methods, 95% did not identify the research paradigm, and 78% did not state the weight given to individual phases. The extent of data integration was 73%, but 83% of studies integrated data using narrative summaries with integration occurring at the interpretation (69.8%). Few studies used joint displays (10.9%), transformation (3.1%), and triangulation (1.6%) for data integration. Mixed-methods research is still in its infancy in nursing, and researchers encounter challenges during its conduct, analysis, and reporting. There is a need to determine researchers' attitudes and challenges toward using mixed methods and educate them about advanced mixed methods. Emphasis should be placed on use of advanced data integration methods so that the rigor and quality of mixed research can be enhanced in nursing research.


Predictive value of platelet to lymphocyte ratio and neutrophil to lymphocyte ratio in evaluating both lung involvement and severity of patients with coronavirus disease 2019.
To investigate the relationship of the neutrophil lymphocyte ratio (NLR) and the platelet lymphocyte ratio (PLR) with lung involvement and total lung severity score (TLSS) in computed tomography (CT) of patients with coronavirus disease -19 (COVID-19) and to evaluate their clinical usability. Basic laboratory, clinical features and imaging data of patients was obtained by examining the file and archive records of our hospital. According to the findings of lung CT scan at the time of diagnosis among COVID-19 patients, 2 groups were formed. The NLR was 2.22±11.15 and the PLR was 142.77±387.10 in patients with COVID-19 pneumonia. The NLR was 1.88±7.47 and the PLR was 130.65±203.6 8 in patients without COVID-19 pneumonia. The differences in the NLR and the PLR were determined to be statistically significant between the 2 groups. A positive correlation was observed between NLR and PLR (r=0.225, p=0.010) and TLSS (r=0.244, p=0.005). This study showed that the NLR and PLR values can be 2 inflammatory markers that can be used to evaluate lung involvement and disease severity in COVID-19 patients. At the time of initial diagnosis and during follow-up, these markers can give an idea in terms of prognosis, together with other clinical findings and markers.


Idecabtagene Vicleucel Is Well Tolerated and Effective in Relapsed/Refractory Myeloma Patients with Prior Allogeneic Stem Cell Transplantation.
BCMA-specific chimeric-antigen receptor (CAR-) T cell therapy has led to high response rates and durable remissions in patients with relapsed refractory multiple myeloma. However, little data are available for patients after prior allogeneic stem cell transplantation (allo-SCT) in whom T cells are chimeric. In this study, we aimed to assess the safety and efficacy of patient-derived donor CAR-T therapy in myeloma patients with prior allo-SCT, particularly with regard to graft-versus-host disease (GVHD). We report a comprehensive clinical analysis of 3 patients who had previously undergone allo-SCT for high-risk myeloma and were treated with idecabtagene vicleucel (ide-cel) at our institution. Ide-cel was well tolerated, with no clinically relevant immune effector cell-associated neurotoxicity syndrome or cytokine release syndrome observed in any patient. Importantly, no new GVHD was observed, even though all patients had a history of GVHD. All patients responded to treatment with at least a very good partial remission. Two patients relapsed within 6 months, and 1 patient was still in stringent complete remission at the time of this report. Our findings demonstrate that treatment with ide-cel is feasible, very well tolerated, and effective in patients with relapsed/refractory multiple myeloma after prior allo-SCT.


Cyclopropyl-containing positive allosteric modulators of metabotropic glutamate receptor subtype 5.
Positive allosteric modulators (PAMs) binding to the transmembrane (TM) domain of metabotropic glutamate receptor 5 (mGluR5) are promising therapeutic agents for psychiatric disorders and traumatic brain injury (TBI). Novel PAMs based on a trans-2-phenylcyclopropane amide scaffold have been designed and synthesized. Facilitating ligand design and allowing estimation of binding affinities to the mGluR5 TM domain was the novel computational strategy, site identification by ligand competitive saturation (SILCS). The potential protective activity of the new compounds was evaluated using nitric oxide (NO) production in BV2 microglial cell cultures treated with lipopolysaccharide (LPS), and the toxicity of the new compounds tested using a cell viability assay. One of the new compounds, 3a, indicated promising activity with potency of 30 μM, which is 4.5-fold more potent than its lead compound 3,3'-difluorobenzaldazine (DFB), and showed no detectable toxicity with concentrations as high as 1000 μM. Thus this compound represents a new lead for possible development as treatment for TBI and related neurodegenerative disorders.


Validation of the SafTest Peroxide Test Kit for the Measurement of the Peroxide Content of Oils, Tallows, Meat Meals, Potato Chips, and Grain-Based Snacks: AOAC Performance Tested MethodSM 101001.
Peroxide value is used as an indicator of oxidative degradation, off flavor, and rancidity in fat. Traditional methods are dependent on a fat extraction step to extract fat for subsequent titration of peroxides. The SafTest Peroxide Test offers a direct measurement of the peroxide value of vegetable oils, animal fats, meat meal and fish meal products, and potato chips, crackers, and other processed grain-based snack products using micro-analytical and membrane separation principles. The study objective is to validate the SafTest Peroxide Test in an internal study, two contracted studies, and an independent laboratory study. Calibration, LOQ, robustness, selectivity, matrix, shelf life, and lot-to-lot stability were evaluated. The practical dynamic range of the calibration is 0.00 to 0.20 meq/kg. LOQ was determined as 0.02 meq/kg for liquids analyzed neat and 0.08 to 0.46 meq/kg for dry matrixes requiring a 1:4 to a 1:8 dilution. SafTest Peroxide Test relative standard deviations of repeatability (RSDr) averaged 1-13% compared to AOCS Cd 8-53 RSDr% of 1-15% and demonstrated average recoveries of 95.7, 91.5, and 105.2% of the AOCS Cd 8-53 results for olive oil, meat meal and fish, and animal fat and oils, respectively. The SafTest Peroxide Test is an acceptable alternative to AOCS Cd 8-53 and AOCS Cd 8 b-90 for the measurement of peroxide value in oils, fats, meat meal, fish meal, and snack products. The SafTest Peroxide Test uses small reagent volumes, eliminates fat extraction, and provides rapid and standardized procedures for the detection of peroxide.


Mothers' experiences with mHealth intervention for postnatal care utilisation in Nigeria: a qualitative study.
The postnatal period implies a crucial and delicate time for both the mother and the newborn. There is a growing body of evidence that is increasingly pointing to mHealth interventions as a potential tool for improved utilisation of maternal and child health services, including postnatal care. This can promote the health of mother and baby during this delicate period. However, the success of the interventions must be explored to validate their usefulness and reliability. Hence, this study explored the experiences of postpartum women on the usefulness of the mHealth intervention (postnatal care assistant) they received. Twenty women, who were involved in mHealth intervention were interviewed using a semi-structured interview guide. They were recruited from the intervention group of a quasi-experimental study that evaluated the effect of a mHealth intervention on the uptake of postnatal care services. Thematic analysis of data was done using NVivo software version 10. Five major themes emerged from data shared by the participants. They are general feelings about the messages, benefits derived from the messages about pregnancy and hospital delivery, increased knowledge about baby care, facilitation of PNC utilisation and involvement of significant others in decision making. They affirmed that the information and reminder messages gave them the impetus to utilise postnatal care services. Mothers reported that mHealth intervention provided immense support and assistance during pregnancy and the reminder messages encouraged them to utilise postnatal care services. This study suggests that improved education and reminder messages via mobile phones are needed during pregnancy and after childbirth to promote mother and child health through the utilization of postnatal care services, and efforts to put this approach to action should be pursued.


Better informed: an overview of health informatics.
Healthcare informatics is increasing in importance both for healthcare administrators and medical and dental practitioners. Governments across the developed world are initiating major national health IT programmes. At the same time, future best medical and dental practice will increasingly depend on computer-based support tools, although disagreement remains about the effectiveness of current support tools. Over the longer term, future informatics tools, combined with other medical and dental technology modalities, promise more adaptive, patient-focused and efficient healthcare and education for the practitioner and the patient.


Cerebral blood flow changes during tilt table testing in healthy volunteers, as assessed by Doppler imaging of the carotid and vertebral arteries.
Using different techniques, reduction of cerebral blood flow (CBF) during orthostatic stress were demonstrated. One study reported flow reduction of the right internal carotid (ICA) and vertebral (VA) artery during orthostatic stress by Doppler imaging, with different effects on the 2 vessels. Global CBF changes, using this technique, have not been reported. Therefore, flow of the ICA, VA and global CBF were measured during head-up tilt testing. 33 healthy volunteers underwent tilt testing. At three time points (supine, half way and at the end of the test) Doppler imaging of the ICA and VA was performed, as well as PetCO2 measurements. Global CBF was significantly reduced by 4.5 ± 2.8% halfway the test and by 6.0 ± 3.4% at the end. All 4 artery flows were significantly reduced during the tilt, without differences between them. Despite small changes in PetCO2 there was a significant relation between de CBF decrease and PetCO2 decrease (p < 0.05). Orthostatic stress in HV results in a small but significant reduction of CBF by a homogenous reduction in the four cerebral vessels and is modulated by PetCO2 changes. CBF changes can be measured during tilt testing using Doppler VA and ICA imaging.


Mapping of Sp1 regulation sites in the promoter of the human alpha1-proteinase inhibitor gene.
Keratoconus is a potentially blinding disease that thins the central cornea. In afflicted corneas, the level of an inhibitor, alpha1-proteinase inhibitor (alpha1-PI), is found reduced. An increased expression of transcription factor Sp1 is also demonstrated. To examine the role of Sp1 in regulation of the human alpha1-PI gene, a 1.4-kb (-1397/+9) 5'-flanking promoter sequence that contains 10 Sp1 sites was cloned. Previous transient transfection experiments showed that Sp1 expression indeed repressed the alpha1-PI promoter activity. In this study, 12 DNA segments, a series of 5', 3', and internal deletions of the 1.4-kb alpha1-PI promoter sequence, were ligated into the SEAP (secreted alkaline phosphatase) reporter gene vector and transfected into human corneal stromal cells. Co-transfection with a Sp1 expression vector pPacSp1 was also performed in parallel. The SEAP enzyme activity was assayed. A fragment with 489 bp (-480/+9) of the 3' sequence, and three fragments with internal deletions, were found to confer a majority of the full promoter activity. Other deletions significantly abolished the promoter activity. Site-directed mutagenesis experiments further revealed that the most proximal Sp1 site (-100/-87) may be an essential element involved in the negative regulation of alpha1-PI promoter activity by Sp1. Interaction between the proximal and distal Sp1 sites also seemed to be important. These results provide the first in-depth characterization of the transcription mechanisms regulating the expression of alpha1-PI. Mapping of the Sp1 sites may help elucidate the molecular pathway leading to the alterations observed in keratoconus.


Spermatogenesis after gonadotoxic childhood treatment: follow-up of 12 patients.
What is the long-term impact of presumed gonadotoxic treatment during childhood on the patient's testicular function at adulthood? Although most patients showed low testicular volumes and some degree of reproductive hormone disruption 12.3 (2.3-21.0) years after gonadotoxic childhood therapy, active spermatogenesis was demonstrated in the semen sample of 8 out of the 12 patients. In recent decades, experimental testicular tissue banking programmes have been set up to safeguard the future fertility of young boys requiring chemo- and/or radiotherapy with significant gonadotoxicity. Although the risk of azoospermia following such therapies is estimated to be high, only limited long-term data are available on the reproductive potential at adulthood. This single-centre prospective cohort study was conducted between September 2020 and February 2023 and involved 12 adult patients. This study was carried out in a tertiary care centre and included 12 young adults (18.1-28.3 years old) who had been offered testicular tissue banking prior to gonadotoxic treatment during childhood. All patients had a consultation and physical examination with a fertility specialist, a scrotal ultrasound to measure the testicular volumes and evaluate the testicular parenchyma, a blood test for assessment of reproductive hormones, and a semen analysis. Testicular tissue was banked prior to the gonadotoxic treatment for 10 out of the 12 included patients. Testicular volumes were low for 9 patients, and 10 patients showed some degree of reproductive hormone disruption. Remarkably, ongoing spermatogenesis was demonstrated in 8 patients at a median 12.3 (range 2.3-21.0) years post-treatment. This study had a limited sample size, making additional research with a larger study population necessary to verify these preliminary findings. These findings highlight the need for multicentric research with a larger study population to establish universal inclusion criteria for immature testicular tissue banking. This study was conducted with financial support from the Research Programme of the Research Foundation-Flanders (G010918N), Kom Op Tegen Kanker, and Scientific Fund Willy Gepts (WFWG19-03). The authors declare no competing interests. NCT04202094; https://clinicaltrials.gov/ct2/show/NCT04202094?id=NCT04202094&draw=2&rank=1 This study was registered on 6 December 2019, and the first patient was enrolled on 8 September 2020.


Systematic understanding the mechanisms of vitiligo pathogenesis and its treatment by Qubaibabuqi formula.
Vitiligo is a depigmentation disorder, which results in substantial cosmetic disfigurement and poses a detriment to patients' physical as well as mental. Now the molecular pathogenesis of vitiligo still remains unclear, which leads to a daunting challenge for vitiligo therapy in modern medicine. Herbal medicines, characterized by multi-compound and multi-target, have long been shown effective in treating vitiligo, but their molecular mechanisms of action also remain ambiguous. Here we proposed a systems pharmacology approach using a clinically effective herb formula as a tool to detect the molecular pathogenesis of vitiligo. This study provided an integrative analysis of active chemicals, drug targets and interacting pathways of the Uygur medicine Qubaibabuqi formula for curing Vitiligo. The results show that 56 active ingredients of Qubaibabuqi interacting with 83 therapeutic proteins were identified. And Qubaibabuqi probably participate in immunomodulation, neuromodulation and keratinocytes apoptosis inhibition in treatment of vitiligo by a synergistic/cooperative way. The drug-target network-based analysis and pathway-based analysis can provide a new approach for understanding the pathogenesis of vitiligo and uncovering the molecular mechanisms of Qubaibabuqi, which will also facilitate the application of traditional Chinese herbs in modern medicine.


Two mosaic terminal inverted duplications arising post-zygotically: Evidence for possible formation of neo-telomeres.
To elucidate the structure of terminal inverted duplications and to investigate potential mechanisms of formation in two cases where there was mosaicism with cells of apparently normal karyotype. A karyotype [46,XY,inv dup(4)(p16.3p15.1)/46,XY] performed on blood lymphocytes from a patient referred for developmental delay (case 1) demonstrated a normal karyotype in 60% of cells with a terminal inverted duplication 4p in the remainder. In case 2, referred for multiple fetal anomalies on an ultrasound scan, 33% of amniocyte colonies were karyotypically normal, with a terminal inv dup 10p in the remainder [46,XX,inv dup(10)(p15.3p11)/46,XX]. Duplicated FISH signals for GATA3 and NEBL loci (in case 2), and for the Wolf-Hirschhorn locus (case 1) confirmed the inverted structure of both duplications. In the GTL banded normal cells from both cases, there was a cryptic deletion detected by FISH of one copy of the subtelomere 4p (case 1, probe GS-36P21), and subtelomere 10p (case 2, probe GS-306F7). At pter on both inv dup chromosomes there was no FISH signal present for the specific subtelomere probe. However, a positive pantelomeric probe signal was detected at 4 pter and 10 pter in both the cryptically-deleted chromosomes and the inv dup chromosomes in the respective cell lines of both cases. An inv dup structure was evident for both cases on GTL bands, and confirmed by the various FISH studies. The presence of telomere (TTAGGG repeat) sequences at pter on the inv dup chromosomes (where more proximal chromosome specific subtelomeric probes were negative) was indicated by the pantelomeric probe signals in both cases. We conclude the most likely mechanism of origin in both cases was by sub-telomeric breakage in the zygote at pter, and delayed repair/rearrangement until after one or more subsequent mitotic divisions. In these divisions, at least one breakage-fusion-bridge cycle occurred, to produce inverted duplications. It is proposed then that two differently "repaired" daughter cells proliferated in parallel. In one daughter cell line (with an overtly normal karyotype) there was deletion of the subtelomere and presumed repair through capping by a neo-telomere (i.e. "healing", as initially proposed by McClintock). This occurred in both cases presented. In the other daughter cell of each case, it is proposed that chromosome stabilization was achieved (after replication) by sister chromatid reunion to form a dicentric, which broke at a subsequent anaphase, to form an inverted duplication (with loss of the reciprocal product, and the other daughter cell line). One inv dup was repaired without an interstitial specific subtelomere (case 1) and one was repaired with a duplicated specific interstitial subtelomere (case 2). After repair TTAGGG repeats were detected by FISH at each respective new pter.


Synthesis of N-(1-ethyl-4-methylhexahydro-1,4-diazepin-6-yl)nicotinamides and their affinities for 5-HT3 and dopamine D2 receptors.
A series of N-(1-ethyl-4-methylhexahydro-1,4-diazepin-6-yl)nicotinamide derivatives were prepared and evaluated for their binding to 5-HT3 and dopamine D2 receptors. Among them, the 5-bromo-2-methoxy-6-methylaminonicotinamide 16 and its (R)-isomer were found to have potent affinities for both receptors. The affinities of (R)-16 for 5-HT3 and dopamine D2 receptors are approximately 3-fold higher than those of the corresponding benzamide (R)-1 (IC50: 1.1 and 12 nM vs. 2.9 and 35 nM, respectively).


Electron microscopy of fine-needle aspiration biopsy from extragonadal germ cell tumors.
We describe five cases of extragonadal germ cell tumor (EGCT) diagnosed by the electron microscope (EM) on cytological material. The clinical diagnosis was incorrect in all cases and EGCT was suspected in two cases; cytological diagnosis by light microscopy confirmed the presence of malignant tumor cells, but did not identify the cytotype/s correctly except in one case. Ultrasonography, laparoscopy, and autopsy (in case 3) excluded a primitive germ cell tumor (GCT). Histology confirmed the EM diagnosis in all cases. EM, even of scanty or necrotic cytological material, is particularly useful for mediastinal and retroperitoneal masses. In case of EGCT, EM can identify the different cytotypes and the different ultrastructural subcellular cytotypes and demonstrates a close relation between seminomatous and nonseminomatous GCT, which could influence their classification and prognosis.


OnDA: online data analysis and feedback for serial X-ray imaging.
This article describes a free and open-source data analysis utility designed for fast online feedback during serial X-ray diffraction and scattering experiments: OnDA (online data analysis). Three complete real-time monitors for common types of serial X-ray imaging experiments are presented. These monitors are capable of providing the essential information required for quick decision making in the face of extreme rates of data collection. In addition, a set of modules, functions and algorithms that allow developers to modify the provided monitors or develop new ones are provided. The emphasis here is on simple, modular and scalable code that is based on open-source libraries and protocols. OnDA monitors have already proven to be invaluable tools in several experiments, especially for scoring and monitoring of diffraction data during serial crystallography experiments at both free-electron laser and synchrotron facilities. It is felt that in the future the kind of fast feedback that OnDA monitors provide will help researchers to deal with the expected very high throughput data flow at next-generation facilities such as the European X-ray free-electron laser.


A cellular threshold for active ERK1/2 levels determines Raf/MEK/ERK-mediated growth arrest versus death responses.
In addition to its conventional role for cell proliferation and survival, the Raf/MEK/Extracellular signal-regulated kinase (ERK) pathway can also induce growth arrest and death responses, if aberrantly activated. Here, we determined a molecular basis of ERK1/2 signaling that underlies these growth inhibitory physiological outputs. We found that overexpression of ERK1 or ERK2 switches ΔRaf-1:ER-induced growth arrest responses to caspase-dependent apoptotic death responses in different cell types. These death responses, however, were reverted to growth arrest responses upon titration of cellular phospho-ERK1/2 levels by the MEK1/2 inhibitor AZD6244. These data suggest that a cellular threshold for active ERK1/2 levels exists and affects the cell fate between death and growth arrest. We also found that death-mediating ability of ERK2 is abolished by the catalytic site-disabling Lys52Arg replacement or significantly attenuated by the F-site recruitment site-disabling Tyr261Asn replacement, although unaffected by the mutations that disable the common docking groove or the dimerization interface. Therefore, ERK1/2 mediates death signaling dependently of kinase activity and specific physical interactions. Intriguingly, Tyr261Asn-replaced ERK2 could still mediate growth arrest signaling, further contrasting the molecular basis of ERK1/2-mediated growth arrest and death signaling. These data reveal a mechanism underlying the role of ERK1/2 as a focal point of Raf/MEK/ERK-mediated growth arrest and death signaling.


The Genomic Characterization of Two Microbacterium foliorum-Specific Bacteriophages, QuadZero and AnnaLie.
The microbacteriophages QuadZero and AnnaLie were isolated from soil samples from Charlotte, NC, and were classified into EA and EB clusters, respectively. QuadZero has a 40,140 base-pair double-stranded DNA genome with 62 predicted protein coding genes, whereas AnnaLie has a 41,665-bp genome with 71 predicted protein coding genes.


Aortic stiffness and diastolic flow abnormalities in end-stage renal disease assessed by magnetic resonance imaging.
Arterial stiffness is associated with adverse cardiovascular outcomes, particularly in end-stage renal disease (ESRD). One mechanism linking arterial stiffness with cardiovascular events may be the changes in pressure wave reflection on ventricular ejection and coronary perfusion during diastole. We illustrate this using MRI to describe aortic elastic properties and alterations of diastolic flow in comparison to derived central pressure characteristics. Ten patients with ESRD and ten control subjects were studied. Transverse images of the ascending aorta were obtained by cardiac MRI. Aortic distensibility was calculated using brachial pulse pressure. MRI flow maps were obtained from the ascending aorta and aortic pressure was calculated using SphygmoCor. ESRD patients had reduced aortic distensibility compared to the controls (median 0.00464 mm Hg(-1) vs. 0.00152 mm Hg(-1), p = 0.0057). Furthermore, in diastole, normal subjects show net reversal of blood flow in the ascending aorta, with a mean of -19.6 versus +7.6 ml/min in the ESRD group; p = 0.045. Using non-invasive methods we have demonstrated a marked reduction in aortic distensibility along with disturbances in aortic flow, providing insight into the pathophysiology of ventricular-vascular interaction. The normal group showed reversal of diastolic blood flow, which may have a direct relationship with coronary perfusion parameters, which was absent in the ESRD group.


What Is Different in the Population of the Brazilian Amazon Region so that They Have a Low Frequency of KRAS Gene Mutations?
Colorectal cancer (CRC) has been described in the medical literature as resulting from many forms of interaction between lifestyle, genetics, and geographical origin. Genetically, the KRAS gene has a negative impact on the general survival and prognosis of patients when mutated. This study was conducted in Brazil and included information about 60 patients with CRC stage III and IV examined at the Day Hospital Oncológica do Brasil, whose DNA was analyzed with the PCR-DNA method to determine the existence of a KRAS mutation. The results revealed that 18.3% of the individuals analyzed showed a KRAS mutation (24% of mutations were found in men and 14.3% in women), which is a smaller proportion than those found in other large studies across the globe. As our analysis is one of the very first published about this topic, more studies are necessary to understand the role of KRAS mutations and the associated variables in populations such as that of the Amazon region.


Aerogels from copper (II)-cellulose nanofibers and carbon nanotubes as absorbents for the elimination of toxic gases from air.
A novel deodorizer that is capable of selectively eliminating the odorous chemicals, such as ammonia, trimethylamine, hydrogen sulfide and methyl mercaptan, is described. The deodorizer is a nanostructured aerogel by nature, consisting of 2,2-6,6-tetramethylpiperidine-1-oxyl (TEMPO) oxidized cellulose nanofibrils (CNF), transition metal divalent cations (M2+), and multi-walled carbon nanotubes (CNT) as the constitutive elements. CNF are firstly mixed with M2+ (M2+, in this paper, typifies Ni2+, Co2+ and Cu2+) to form CNF-M2+ complexes, monodispersed CNT is then mixed to prepare CNT/CNF-M2+ waterborne slurries; CNT/CNF-M2+ hybridized aerogels are finally obtained via freezing-drying of the CNT/CNF-M2+ waterborne slurries. The CNT/CNF-M2+ aerogels are a foam-like structure consisting of CNF and CNT as backbones and M2+ as linkers. The aerogels show higher capabilities (in comparison with activated carbon) for selectively adsorbing ammonia, trimethylamine, hydrogen sulfide and methyl mercaptan. Computing simulations suggest a theoretical conclusion that the odorous chemicals are absorbed in a preferring manner of bimolecular absorptions via the M2+ moieties. The CNT/CNF-M2+ hybridized aerogels are lightweight, eco-friendly, and easy to produce in industrial scales. Our new finding, as is described in this paper, demonstrates potential applications of the TEMPO-oxidized CNF to the field of deodorizations.


Glucosides from Vitex agnus-castus.
The methanolic extract of the flowering stems of Vitex agnus-castus yielded three new iridoids: 6'-O-foliamenthoylmussaenosidic acid (agnucastoside A), 6'-O-(6,7-dihydrofoliamenthoyl)mussaenosidic acid (agnucastoside B) and 7-O-trans-p-coumaroyl-6'-O-trans-caffeoyl-8-epiloganic acid (agnucastoside C) in addition to four known iridoids (aucubin, agnuside, mussaenosidic acid and 6'-O-p-hydroxybenzoylmussaenosidic acid) and one known phenylbutanone glucoside (myzodendrone). The structure elucidations were mainly done by spectroscopic methods (1D and 2D NMR spectra) and MS data interpretation. The purified compounds were tested for biological activities against various microorganisms and cancer cell lines.


The predictive value of stimulation index calculated by modified mixed lymphocyte culture in the detection of GVHD following hematopoietic stem cell transplantation.
Mixed lymphocyte culture (MLC) is one of the routine tests performed prior to hematopoietic stem cell transplantation (HSCT) as a predictive assay for assessing the quality of donor matching and graft-versus-host disease (GVHD). The stimulation index is one of the formulas of the MLC test, and it is used for evaluation of matching between donor and recipient. Modified MLC (mMLC) test is produced by adding various cytokines to the MLC test, and increased sensitivity has been reported with this modification. The importance of the stimulation index values in MLC and mMLC tests was evaluated in 59 patients who received HSCs from human leukocyte antigen-identical sibling donors. In the mMLC test, cytokines were added as interleukin (IL)-2, IL-2 + IL-4 and IL-2 + interferon (IFN)-gamma + tumor necrosis factor (TNF)-alpha. Stimulation index values in mMLC test were compared with stimulation index values in MLC test. Twenty-three (39%) patients developed GVHD. When evaluated in terms of stimulation index >1 patients, in MLC, 55% of the patients developed GVHD (p=0.229), whereas these values were 75% in the IL-2 added mMLC test (p=0.035), 100% in the IL-2 + IL-4 added mMLC test (p=0.076) and 85.7% in the IL-2 + IFN-gamma + TNF-alpha added mMLC test (p=0.015). mMLC increased the sensitivity of the test. The relation between the positive results and evidence of GVHD after transplantation was found significant.


Tris(ethane-1,2-di-amine-κ2 N,N')zinc(II) tetra-chlorido-zincate(II).
The title complex, [Zn(C2H8N2)3][ZnCl4], exists as discrete ions. The [Zn(C2H8N2)3]2+ cation exhibits a distorted octa-hedral shape. In the [ZnCl4]2- anion, the ZnII atom is in an almost regular tetra-hedral environment. The crystal packing is consolidated by N-H⋯Cl and C-H⋯Cl hydrogen bonds.