Using microarrays to study the microenvironment in tumor biology: the crucial role of statistics.
Microarrays represent a potentially powerful tool for better understanding the role of the microenvironment on tumor biology. To make the best use of microarray data and avoid incorrect or unsubstantiated conclusions, care must be taken in the statistical analysis. To illustrate the statistical issues involved we discuss three microarray studies related to the microenvironment and tumor biology involving: (i) prostatic stroma cells in cancer and non-cancer tissues; (ii) breast stroma and epithelial cells in breast cancer patients and non-cancer patients; and (iii) serum associated with wound response and stroma in cancer patients. Using these examples we critically discuss three types of analyses: differential gene expression, cluster analysis, and class prediction. We also discuss design issues.


Preliminary study of icariin indicating prevention of steroid-induced osteonecrosis of femoral head by regulating abnormal expression of miRNA-335 and protecting the functions of bone microvascular endothelial cells in rats.
The pathogenesis of osteonecrosis of the femoral head (ONFH) is unclear. Our previous study demonstrated that upregulated miR-335 in bone microvascular endothelial cells (BMECs) might be associated with the disease of steroid-induced ONFH. Here, we study the preventive effect of ICA on steroid-induced ONFH in rats. 90 rats were separated into three groups: control group, methylprednisolone (MPS) group, and MPS + Icariin (ICA) group. Four weeks later, histological analyses were performed. Thrombomodulin (TM) and vascular endothelial growth factor (VEGF) were tested. MiRNA-335 expression was screened in the three groups using Agilent Gene Spring GX software. Target genes of miRNA-335 were detected by bioinformatics analysis. The functions of BMECs were analyzed by scratch, angiogenesis and cell survival rate. ICA can prevent the occurrence of steroid-associated ONFH in rats and reduce the amount of TM and VEGF in serum induced by glucocorticoids. ICA could regulate the overexpression of miRNA-335 induced by glucocorticoids. We predicted the Gene ontology (GO) and signaling pathways of target genes. At 24 hours, we found that ICA significantly promoted BMECs migration abilities. We also found that ICA could promote the angioplasty ability of BMECs. ICA could improve the survival rate of BMECs after steroid-induced injury. ICA is effective to prevent the occurrence of steroidinduced ONFH. ICA has a protective effect against steroid-induced BMECs injury. ICA regulated the imbalance of miRNA-335 expression induced by the glucocorticoid in BMECs, which provides a new viewpoint to explore the mechanism of ICA in preventing steroid-induced ONFH.


Hemodiafiltration using dialysate as substitution fluid.
To reduce the cost of hemodiafiltration (HDF), a standard hemodialysis machine was modified to permit in-line production of substitution fluid from bicarbonate dialysate. The authors' present data showing that this procedure was made bacteriologically safe; was simple to use, and offered a cheap approach to perform high-flux HDF using a hemodifilter of relatively small membrane surface area.


Characterization of Plasmodium falciparum cloned lines with respect to gametocyte production in vitro, infectivity to Anopheles mosquitoes, and transmission to Aotus monkeys.
The production of gametocytes in vitro and their subsequent infectivity to mosquitoes by 3 cloned lines of Plasmodium falciparum were studied. 2 of the cloned lines, Honduras I-clone B3 and Indochina III-clone W2, produced mature gametocytes (stage V) that were infective to Anopheles mosquitoes. The third clone, Sierra Leone I-clone D6, produced gametocytes, the majority of which did not develop beyond stage III. Fully mature gametocytes of Sierra Leone I-clone D6 were not infective to mosquitoes. Sporozoites collected from An. freeborni infected with Honduras I-clone B3 were used in transmission studies. Two of three Aotus monkeys were infected after prepatent periods of 19 and 20 d, respectively. This study supports previous reports that cloned lines of P. falciparum contain the full genetic capacity to produce morphologically mature gametocytes. The transmission to Aotus monkeys has also conclusively established that biologically competent gametocytes of both sexes are produced by clones.


A membrane-permeant, bioactivatable derivative of Ins(1,3,4)P3 and its effect on Cl(-)-secretion from T84 cells.
The synthesis of rac-2,5,6-tri-O-butyryl-myo-inositol 1,3,4-trisphosphate hexakis(acetoxymethyl) ester [Bt3-Ins(1,3,4)P3/AM, 1], a membrane-permeant derivative of myo-inositol 1,3,4-trisphosphate [Ins(1,3,4)P3] is reported. 1 inhibited calcium-mediated chloride secretion of T84 cells, suggesting a regulatory link of Ins(1,3,4)P3 and the biosynthesis of the known inhibitor myo-inositol 3,4,5,6-tetrakisphosphate.


Clinical Application of Melatonin in the Treatment of Cardiovascular Diseases: Current Evidence and New Insights into the Cardioprotective and Cardiotherapeutic Properties.
Cardiovascular diseases (CVDs) are the leading global cause of mortality and disability, tending to happen in younger individuals in developed countries. Despite improvements in medical treatments, the therapy and long-term prognosis of CVDs such as myocardial ischemia-reperfusion, atherosclerosis, heart failure, cardiac hypertrophy and remodeling, cardiomyopathy, coronary artery disease, myocardial infarction, and other CVDs threatening human life are not satisfactory enough. Therefore, many researchers are attempting to identify novel potential therapeutic methods for the treatment of CVDs. Melatonin is an anti-inflammatory and antioxidant agent with a wide range of therapeutic properties. Recently, several investigations have been carried out to evaluate its effectiveness and efficiency in CVDs therapy, focusing on mechanistic pathways. Herein, this review aims to summarize current findings of melatonin treatment for CVDs.


Relevance of ovarian signaling for the early behavioral transition from estrus to pregnancy in the female rabbit.
During estrus, the female domestic rabbit (Oryctolagus cuniculus) displays scent marking behavior (chinning), which is immediately inhibited after mating, temporarily recovers, and then declines and remains inhibited across pregnancy. Chinning is inhibited by progesterone (P) and the activation of the progesterone receptor (PR), but it is unlikely that P participates in the "acute" (immediate) or "early" inhibition of chinning (24 to 96 h post-mating, before plasma P levels rise). Since PR is activated in a ligand-independent manner by a variety of signaling molecules, some of which (e.g., GnRH) are also associated with reflexive ovulation in this species, we hypothesized that neurochemical/neuroendocrine signals associated with mating activate PR, resulting in the inhibition of chinning. In Experiment 1, we tested whether the PR antagonist, RU486 (20 mg, injected s.c. at -1 h, or at -7 h and +3 h relative to mating) prevented the post-mating inhibition of chinning in intact females. RU486 did not prevent the post-mating decline in chinning, indicating that PR activation associated with mating is not necessary for this effect. In Experiment 2, we used ovariectomized (OVX), estradiol benzoate (EB)-treated females to test the hypothesis that ovarian signaling is necessary for the post-mating inhibition of chinning. The acute inhibition of chinning occurred in OVX females, but the early inhibition was absent. We conclude that ovarian signaling is necessary for the early, but not acute, post-mating inhibition of chinning. The PR seems not to participate in either of these phases.


[Type II CRISPR-Cas System Nucleases: A Pipeline for Prediction and In Vitro Characterization].
The use of CRISPR-Cas bacterial adaptive immunity system components for targeted DNA changes has opened broad prospects for programmable genome editing of higher organisms. The most widely used gene editors are based on the Cas9 effectors of the type II CRISPR-Cas systems. In complex with guide RNAs, Cas9 proteins are able to directionally introduce double-stranded breaks into DNA regions that are complementary to guide RNA sequences. Despite the wide range of characterized Cas9s, the search for new Cas9 variants remains an important task, since the available Cas9 editors have several limitations. This paper presents a workflow for the search for and subsequent characterization of new Cas9 nucleases developed in our laboratory. Detailed protocols describing the bioinformatical search, cloning, and isolation of recombinant Cas9 proteins, testing for the presence of nuclease activity in vitro, and determining the PAM sequence, which is required for recognition of DNA targets, are presented. Potential difficulties that may arise, as well as ways to overcome them, are considered.


A randomized trial of tacrolimus versus tacrolimus and prednisone for the maintenance of disease remission in noninfectious uveitis.
To compare tacrolimus monotherapy with tacrolimus and prednisone therapy for the maintenance of disease remission in subjects with noninfectious posterior segment intraocular inflammation (PSII). Randomized, controlled, phase 2b, open-label, dual-center noninferiority trial. Fifty-eight patients with sight-threatening PSII. Patients requiring a second-line systemic immunosuppressive agent to control their PSII were treated with therapeutic doses of oral tacrolimus. Those subjects who subsequently were able to taper their prednisone dose to 10 mg daily without disease reactivation were assigned randomly either to stop prednisone or to continue 7.5 to 10 mg prednisone daily for 9 months. Change in logarithm of the minimum angle of resolution (logMAR) visual acuity (VA) and rate of patient withdrawal resulting from treatment inefficacy or intolerance. Thirty-five patients successfully tapered their prednisone to 10 mg daily. Of these, 16 were allocated randomly to receive tacrolimus monotherapy and 19 to continue taking prednisone and tacrolimus dual therapy. The difference in the mean change in VA for monotherapy compared with the dual therapy group was less than 1 logMAR letter (logMAR, -0.008; 95% confidence interval, -0.108 to 0.092; P = 0.870). The proportion of patients who tolerated treatment and maintained disease remission for 9 months after randomization also was similar in both groups (monotherapy, 62.5%; dual therapy, 68.4%; P = 0.694). All monotherapy treatment failures were the result of disease reactivation, whereas 50% of dual-therapy failures were the result of drug intolerance. This study provides preliminary evidence that corticosteroids can be withdrawn in tacrolimus-treated patients who are able to achieve control of PSII with 10 mg prednisone daily, and any advantage of dual therapy in the prevention of disease reactivation was offset by its greater treatment intolerance. These findings support the further evaluation of corticosteroid-free treatment in future phase 3 trials (International Standard Randomised Controlled Trial Number Register identification, ISRCTN46576063). Proprietary or commercial disclosure may be found after the references.


The serotonin transporter gene in aggressive children with and without ADHD and nonaggressive matched controls.
Using an ethnically homogeneous sample of highly aggressive Caucasian children and their matched controls, we report differential associations of the 5HTTLPR and VNTR polymorphisms with ADHD and aggression, respectively. Given the small sample size and the preliminary nature of our findings, replication is necessary.


Monoterpenoid terpinen-4-ol inhibits voltage-dependent Na+ channels of small dorsal root ganglia rat neurons.
The monoterpenoid terpinen-4-ol (4TERP) is known to inhibit cell excitability, has low toxicity and important pharmacological activities, which are likely related to neural excitability, such as anti-inflammatory, antiepileptic and antinociceptive effects. However, the pharmacological characteristics and mechanisms underlying the effects of 4TERP on blockade of neural action potential are not completely elucidated. Since Na+ current (INa) through voltage-dependent Na+ channels (NaV) is a major mechanism for excitability, the present study investigated the pharmacological characteristics and mechanisms of the action of 4TERP on INa through NaV. For this aim, dissociated small neurons of dorsal root ganglia of adult rats were used for whole cell patch-clamp recordings. 4TERP concentration-dependently inhibits INa (IC50 0.8 ± 0.3 mM; pharmacological efficacy 42.89 ± 5.54%). 4TERP interfered with INa through a mechanism with various components, which includes predominantly channel pore block and sensitivity to frequency of use. In presence of 4TERP (3 mM), decreasing stimulation from 5 Hz to very low frequency (75 s of quiescence previously to stimulation) induced INa decrease to 65.17 ± 5.86% of control. 4TERP also altered (left shift) voltage sensitivity of the steady state activation of NaV. Data are discussed aiming to interpret the importance of blockade of INa through NaV as participant of 4TERP-induced inhibition of membrane excitability.


Matrix-assisted laser desorption/ionization-Fourier-transform ion cyclotron resonance-mass spectrometry analysis of exosomal lipids from human serum.
Exosomes contain biomarkers such as proteins and lipids that help in understanding normal physiology and diseases. Lipids, in particular, are infrequently studied using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) for biomarker discovery. In this study, MALDI was equipped with a high-resolution MS to investigate exosomal lipids from human serum. Exosomal lipids were profiled using MALDI with Fourier-transform ion cyclotron resonance (FTICR)-MS. Four matrices (i.e., α-cyano-4-hydroxycinnamic acid [CHCA], 2,5-dihydroxybenzoic acid, sinapinic acid, and graphene oxide [GO]) and three sample preparation methods (i.e., dried droplet, thin layer, and two layer) were compared for the number of lipid species detected and the relative abundance of each lipid from human serum and human serum exosomes. In sum, 172 and 89 lipid species were identified from human serum and human serum exosomes, respectively, using all the methods. The highest number of exosome lipid species, 69, was detected using the CHCA matrix, whereas only 8 exosome lipid species were identified using the GO matrix. Among the identified lipid species, phosphatidylcholine was identified most frequently, probably due to the use of a positive ion mode. Exosomes and human serum showed comparable lipid profiles as determined using MALDI-FTICR-MS. These findings provide a new perspective on exosomal lipidomics analysis and may serve as a foundation for future lipidomics-based biomarker research using MALDI-FTICR-MS.


The role of portal vein embolization in the surgical management of primary hepatobiliary cancers. A systematic review.
Primary liver and biliary cancers are very aggressive tumors. Surgical treatment is the main option for cure or long term survival. The main purpose of this systematic review is to underline the indications for portal vein embolization (PVE), in patients with inadequate future liver remnant (FLR) and to analyze other parameters such as resection rate, morbidity, mortality, survival after PVE and hepatectomy for primary hepatobiliary tumors. Also the role of trans-arterial chemoembolization (TACE) before PVE, is investigated. A systematic search of the literature was performed in Pub Med and the Cochrane Library from 01.01.1990 to 30.09.2015. Forty articles were selected, including 2144 patients with a median age of 61 years. The median excision rate was 90% for hepatocellular carcinomas (HCCs) and 86% for hilar cholangiocarcinomas (HCs). The main indications for PVE in patients with HCC and presence of liver fibrosis or cirrhosis was FLR <40% when liver function was good (ICGR15 < 10%) and FLR < 50% when liver function was affected (ICGR15:10-20%). The combination of TACE and PVE increased hypertrophy rate and was associated with better overall survival and disease free survival and should be considered in advanced HCC tumors with inadequate FLR. In patients with HCs PVE was performed, after preoperative biliary drainage, when FLR was <40%, in the majority of studies, with very good post-operative outcome. However indications should be refined. PVE before major hepatectomy allows resection in a patient group with advanced primary hepato-biliary tumors and inadequate FLR, with good long term survival.


Hemorrhagic fever with renal syndrome-related encephalopathy: magnetic resonance imaging findings.
To report the incidence and findings of brain magnetic resonance imaging (MRI) in patients with hemorrhagic fever with renal syndrome (HFRS)-related encephalopathy along with its clinical course. Medical records and brain MRI were reviewed from January 2004 to January 2013. The final cohort consisted of 145 patients. Brain MRI findings were correlated with associated clinical stage of disease. The MRI findings associated with clinical course of HFRS-related encephalopathy were posterior reversible encephalopathy syndrome pattern (n=3) in oliguric phase and splenial lesion pattern (n=1) in febrile phase. Brain MRI findings in HFRS-related encephalopathy may be associated with the clinical course of HFRS.


Persistent hyperplastic primary vitreous in an adult: case report with fluorescein angiographic findings.
The clinical features and fluorescein angiographic findings of an 18-year-old man with PHPV are reported. The case is unusual in that the eye survived into adulthood. The lens resorbed spontaneously, thereby permitting study of the retrolental mass by fluorescein angiography. Fluorescein leakage from the persistent vessels was very slow but present, thus indicating a probable disruption of the endothelial tight junctions of the tunica vasculosa lentis.


Very Low Frequencies Maintain Pain Relief From Dorsal Root Ganglion Stimulation: An Evaluation of Dorsal Root Ganglion Neurostimulation Frequency Tapering.
Dorsal root ganglion neurostimulation (DRG-S) is effective in treating various refractory chronic pain syndromes. In preclinical studies, DRG-S at very low frequencies (<5 Hz) reduces excitatory output in the superficial dorsal horn. Clinically, we have also observed the effectiveness of DRG-S at low frequencies. We conducted a case series to describe the effect of very low-frequency DRG-S stimulation on clinical outcomes. DRG-S for refractory low back pain was initiated at parameters consistent with published values. Thereafter, the stimulation frequency of DRG-S was reduced in a stepwise fashion to the lowest frequency that maintained pain relief. Pain intensity, disability, and general health status data were collected at baseline, prior to initiation of tapering, and at four weeks after each patient's lowest effective stimulation frequency was reached. After device activation (N = 20), DRG-S frequency was tapered from 16 to 4 Hz over a 4- to 17-week period, reducing charge-per-second by nearly two-thirds. Even so, pain relief was maintained at more than 75%, with consistent findings in the other measures. DRG-S may have utility in treating chronic pain at lower stimulation frequencies than previously recognized. We have previously theorized that the mechanism of action may involve preferential recruitment of low-threshold mechanoreceptor fibers via the endogenous opioid system. Of clinical relevance, lower frequency stimulation maintains DRG-S efficacy regarding improvements in pain, disability, and quality of life. It can extend battery life and may potentially lead to the development of smaller implantable pulse generators.


Advances in the Radiological Evaluation of and Theranostics for Glioblastoma.
Imaging is essential for evaluating patients with glioblastoma. Traditionally a multimodality undertaking, CT, including CT cerebral blood profusion, PET/CT with traditional fluorine-18 fluorodeoxyglucose (18F-FDG), and MRI have been the mainstays for diagnosis and post-therapeutic assessment. However, recent advances in these modalities, in league with the emerging fields of radiomics and theranostics, may prove helpful in improving diagnostic accuracy and treating the disease.


Does native glenoid anatomy predispose to shoulder instability? An MRI analysis.
It is unclear if native glenohumeral anatomic features predispose young patients to instability and if such anatomic risk factors differ between males and females. The purpose of this study was to compare glenoid and humeral head dimensions between patients with a documented instability event without bone loss to matched controls and to evaluate for sex-based differences across measurements. The authors hypothesized that a smaller glenoid width and glenoid surface area would be significant risk factors for instability, whereas humeral head width would not. A prospectively maintained database was queried for patients aged <21 years who underwent magnetic resonance imaging (MRI) for shoulder instability. Patients with prior shoulder surgery, bony Bankart, or glenoid or humeral bone loss were excluded. Patients were matched by sex and age to control patients who had no history of shoulder instability. Two blinded independent raters measured glenoid height, glenoid width, and humeral head width on sagittal MRI. Glenoid surface area, glenoid index (ratio of glenoid height to width), and glenohumeral mismatch ratio (ratio of humeral head width to glenoid width) were calculated. A total of 107 instability patients and 107 controls were included (150 males and 64 females). Among the entire cohort, there were no differences in glenoid height, glenoid width, glenoid surface area, humeral head width, or glenoid index between patients with instability and controls. Overall, those with instability had a greater glenohumeral mismatch ratio (P = .029) compared with controls. When stratified by sex, female controls and instability patients showed no differences in any of the glenoid or humerus dimensions. However, males with instability had a smaller glenoid width by 3.5% (P = .017), smaller glenoid surface area by 5.2% (P = .015), and a greater glenohumeral mismatch ratio (P = .027) compared with controls. Compared with controls, males with instability were found to have smaller glenoid width and surface area, and a glenoid width that was proportionally smaller relative to humeral width. In contrast, bony glenohumeral morphology did not appear to be a significant risk factor for instability among females. These sex-based differences suggest that anatomic factors may influence risk of instability for male and female patients differently.


Highly potent and selective chiral inhibitors of PDE5: an illustration of Pfeiffer's rule.
A series of potent chiral PDE5 inhibitors are described that are based on the sildenafil architecture but exhibit much greater selectivity over PDE6. Eudismic analysis of the SAR in this series provided a clear illustration of Pfeiffer's rule and indicated that the chiral motif was involved in a highly-stereoselective interaction with PDE5. This PDE5 specificity translated to levels of selectivity over PDE6 that were hitherto unprecedented in the sildenafil scaffold. UK-371,800 (compound 8) was identified as a development candidate from this series that married sildenafil-like molecular properties with high selectivity over PDE6. Clinical data confirm that UK-371,800 has markedly superior human pharmacokinetics to a previously-described higher molecular weight achiral analogue in this template (compound 1).


Agreement of anterior ocular biometric measurements with a new optical biometer and a Scheimpflug tomographer.
To evaluate the agreement between a new optical biometer (AL-Scan) and an anterior-segment Scheimpflug tomographer (Pentacam HR). Eye Hospital of Wenzhou Medical University, Zhejiang, China. Prospective randomized comparative case series. Three consecutive scans of the right eye of each subject were obtained using each device in a random order. The following common parameters were recorded: central corneal thickness (CCT), anterior chamber depth (ACD), aqueous depth, and keratometry (K) readings (steep, flat, and mean). Bland-Altman limits of agreement (LoA) were calculated for the common device parameters. The study comprised 121 healthy subjects (57 men, 64 women) with a mean age of 25.41 years ± 6.86 (SD) (range 18 to 54 years). The mean difference in CCT was 1.61 μm with LoA of -6.29 to 9.52 μm. The mean difference in the aqueous depth and ACD was 0.02 mm with LoA of -0.10 to 0.14 mm and -0.09 to 0.14 mm, respectively. For the 2.4 mm K readings from the new biometer versus the standard K readings from the Scheimpflug tomographer, the mean difference for flat K, steep K, and mean K was 0.10 diopter (D), 0.05 D, and 0.08 D, respectively. The LoA were -0.17 to 0.38 D, -0.29 to 0.39 D, and -0.19 to 0.34 D, respectively. The CCT, ACD, and all K readings were assessed to be clinically interchangeable. There was good agreement between the 2 devices for CCT, ACD, aqueous depth, and K readings. None of the authors has a financial or proprietary interest in any material or method mentioned.


If You're House Is Still Available, Send Me an Email: Personality Influences Reactions to Written Errors in Email Messages.
The increasing prevalence of social media means that we often encounter written language characterized by both stylistic variation and outright errors. How does the personality of the reader modulate reactions to non-standard text? Experimental participants read 'email responses' to an ad for a housemate that either contained no errors or had been altered to include either typos (e.g., teh) or homophonous grammar errors (grammos, e.g., to/too, it's/its). Participants completed a 10-item evaluation scale for each message, which measured their impressions of the writer. In addition participants completed a Big Five personality assessment and answered demographic and language attitude questions. Both typos and grammos had a negative impact on the evaluation scale. This negative impact was not modulated by age, education, electronic communication frequency, or pleasure reading time. In contrast, personality traits did modulate assessments, and did so in distinct ways for grammos and typos.


Does the choice of the neuromuscular blocking agent affect the cardiovascular response to intubation?
Heart rate and arterial pressure changes induced by tracheal intubation, 3 min after administration of atracurium, vecuronium, tubocurarine, pancuronium or alcuronium, have been studied under thiopentonenitrous oxide-oxygen anaesthesia supplemented by either 0.5% halothane or fentanyl 2 micrograms kg-1. Pancuronium and alcuronium were associated with the greatest increase in heart rate and tubocurarine with the greatest decrease in arterial pressure prior to intubation. Following intubation, all groups, with the exception of tubocurarine showed a similar and significant rise in heart and arterial pressure when compared with control values. The cardiovascular response to intubation, particularly the effect on heart rate, was less marked when fentanyl was given at induction and was short lived with atracurium and vecuronium. Although those patients who received tubocurarine showed no significant rise in arterial pressure following tracheal intubation, this was due to significant hypotension occurring in this group prior to intubation.


Running per se stimulates the dendritic arbor of newborn dentate granule cells in mouse hippocampus in a duration-dependent manner.
Laboratory rodents provided chronic unlimited access to running wheels display increased neurogenesis in the hippocampal dentate gyrus. In addition, recent studies indicate that such an access to wheels stimulates dendritic arborization in newly formed neurons. However, (i) the presence of the running wheel in the housing environment might also bear intrinsic influences on the number and shape of new neurons and (ii) the dendritic arborization of new neurons might be insensitive to moderate daily running activity (i.e., several hours). In keeping with these uncertainties, we have examined neurogenesis and dendritic arborization in newly formed granular cells in adult C57Bl/6N male mice housed for 3 weeks under standard conditions, with a locked wheel, with a running wheel set free 3 h/day, or with a running wheel set permanently free. The results indicate that the presence of a blocked wheel in the home cage increased cell proliferation, but not the number of new neurons while running increased in a duration-dependent manner the number of newborn neurons, as assessed by DCX labeling. Morphological analyses of the dendritic tree of newborn neurons, as identified by BrdU-DCX co-staining, revealed that although the presence of the wheel stimulated their dendritic architecture, the amplitude of this effect was lower than that elicited by running activity, and was found to be running duration-dependent.


Treatment of distal radius fractures with intrafocal (kapandji) pinning and supplemental skeletal stabilization.
The technique of intrafocal pinning was originally described by Kapandji. This technique, along with the addition of supplemental external or internal bridging fixation, can be performed quickly with minimal soft tissue damage. This technique is indicated for the treatment of unstable extra-articular or minimally displaced intra-articular fractures of the distal radius. Studies demonstrate that young patients with displaced extra-articular distal radius fractures and minimal comminution can be treated by percutaneous intrafocal pin fixation alone, whereas, patients over 55 years of age and younger patients with comminution involving two or more surfaces of the radial metaphysis (or > 50% of the metaphyseal diameter) require bridging fixation besides percutaneous pin fixation. The use of this technique achieves the goal of surgical treatment of distal radius fractures: restoration of hand and wrist function through the restoration of alignment and articular surface congruity.


Cost-effectiveness of a comprehensive immunization program serving high-risk, uninsured adults.
Despite the numerous social and economic benefits of vaccination, adult immunization rates fall far short of recommended levels costing the United States $9 billion annually in health care expenditures and reduced productivity. While it is well recognized that childhood immunization is highly cost-effective, the economic impact of adult immunization programs varies by disease and is influenced by population demographics. This study aimed to assess the cost-effectiveness of a comprehensive adult immunization program serving high-need populations delivered by a local health department (LHD) in partnership with community organizations. We modeled incremental cost-effectiveness taking the payer perspective of each vaccine separately in simulated cohorts of 100,000 over a 20-year horizon using data provided by the LHD and data from the published literature. We adjusted the results to align with actual program delivery and used them to estimate an incremental cost-effectiveness ratio (ICER) for the entire program. We assessed the effects of varying our base model parameters in univariate sensitivity analyses. We discounted benefits and life years saved (LYS) at 3% and adjusted results to 2016 US$. Four of seven disease models were cost-effective (using a $100,000 CE threshold) with ICERS ranging from $14,260 to $79,022/LYS. Sensitivity analyses did not substantially impact the results. The ICER for program as a whole was $67,940/LYS. A community-delivered comprehensive immunization program serving uninsured, low income, high-risk adults is a cost-effective investment even when most do not receive the full regimen of some vaccines.


Transcriptional expression of six genes located on pBSSB1 of Salmonella enterica serovar Typhi in different growth phases and environmental stresses.
A linear plasmid pBSSB1 of z66-positive Salmonella enterica serovar Typhi was previously identified to be involved in unidirectional flagellar phase variation due to the expression and regulation of fljB(z66) and fljA located on it. However, the expression of the remaining genes and other possible functions of pBSSB1 have not been reported yet. Here, we investigated the transcriptional expression characteristics of six genes (002, 008, 012, 017, 021, and 026) located on the linear plasmid pBSSB1 in different growth phases and environmental stresses by means of qRT-PCR. The results show that these six genes are all transcribed in normal growth condition (LB broth at 37 °C with stirring 250 rpm), and their expression has an increasing trend in general with the growth of the bacteria. Under environmental stresses, there is an increase in expression levels of 002, 008, 012, and 026, while no significant change in expression was observed for 017 and 021. In addition, the expression levels and modes of 017 and 021 are similar in all the growth conditions. To our knowledge, this is the first report describing the transcriptional expression characteristics of these six genes located on pBSSB1 and suggesting that pBSSB1 may be very important in response to the environmental stresses in z66-positive S. enterica serovar Typhi.


Intestinal regeneration by a novel surgical procedure.
Treatment of short bowel syndrome is problematical. Small bowel tissue engineering has achieved modest results in animal studies. The aim of this study was to investigate intestinal regeneration in a novel surgical model. Roux-en-Y bypass procedures were performed on 40 Wistar rats weighing 250-350 g. Animals were killed at 1, 2, 3, 4, 8, 12 and 24 weeks after implantation with a 3-cm silicone tube. The spatio temporal relationship of intestinal regeneration was analysed using three-dimensional multislice computed tomography, and examination of sequential morphological changes on gross or histological findings and measurement of missing intestinal tissue (growth defects). Progressive intestinal regeneration on a silicone tube was identifiable in 35 animals. Most adhesions were initially localized on the tube but spread to a distal site 4 weeks after implantation. Growth defects decreased with time, with a marked reduction in the first 4 weeks and a gradual reduction to week 24 after implantation. Luminal patency shown radiologically as well as sequential histological findings, such as mucosal lining, matrix remodelling and muscular regeneration, suggested that regeneration of intestinal tissue took place, not merely scar contraction. Non-invasive as well as histomorphological assessment followed intestinal regeneration over time in this model, which provides scope for further studies.


Serotype-associated immune response and network immunoclusters in children and adults during acute Dengue virus infection.
The present study was designed as an exploratory investigation to characterize the overall profile of chemokines, growth factors, and pro-inflammatory/regulatory cytokines during acute DENV infection according to DENV-1, DENV-2, DENV-4 serotypes and age: children: <1-10-year-old (yo); adolescents:11-20 yo; adults 21-40 yo; and older adults: 41-75 yo. The levels of soluble immunemediators were measured in serum by high-throughput microbeads array in 636 subjects including 317 DENV-infected and 319 age-matching non-infected control (NI). Overall, most soluble mediators were increased in DENV-infected patients as compared to NI group regardless of age and DENV serotype, with high magnitude order of increase for CCL2, CXCL10, IL-1β, IFN-γ, IL1-Ra (fold change >3x), except PDGF in which no fold change was observed. Moreover, despite the age ranges, DENV-1 and DENV-4 presented increased levels of VEGF, IL-6, and TNF-α in serum but decreased levels of PDGF, while DENV-2 exhibited increased levels of CXCL8, CCL4, and IL-12. Noteworthy was that DENV-2 showed increased levels of IL-12, IL-15, IL-17, IL-4, IL-9, and IL-13, and maintained an unaltered levels of PDGF at younger ages (<1-10 yo and 11-20 yo), whereas in older ages (21-40 yo and 41-75 yo), the results showed increased levels of CCL2, IL-6, and TNF-α, but lower levels of PDGF. In general, DENV infection at younger age groups exhibited more complex network immunoclusters as compared to older age groups. Multivariate analysis revealed a clustering of DENV cases according to age for a set of soluble mediators especially in subjects infected with DENV-2 serotype. Altogether, our findings demonstrate that the profile of circulating soluble mediators differs substantially in acute DENV according to age and DENV serotypes suggesting the participation of serotype-associated immune response, which may represent a potential target for development of therapeutics and could be used to assist medical directive for precise clinical management of severe cases.


Developing a statewide protocol to ensure patients with suspected emergent large vessel occlusion are directly triaged in the field to a comprehensive stroke center: how we did it.
We describe the process by which we developed a statewide field destination protocol to transport patients with suspected emergent large vessel occlusion to a comprehensive stroke center.


Decolorization of wastewater of a Baker's yeast plant by membrane processes.
The aim of this study is to develop a membrane-based treatment scheme to remove colorants from the effluent of a baker's yeast plant. For this purpose microfiltration (MF), ultrafiltration (UF) and nanofiltraton (NF) membranes with differing molecular weight cut-offs (MWCOs) were tested. To evaluate the effectiveness of membrane processes in treating the waste stream, optical density (OD), COD, color measurements together with permeation fluxes were used. Effects of pretreatment methods (coagulation and coarse filtration) and feed composition on OD, color, COD were studied. In addition, gel filtration analysis was employed to characterize feed and permeate streams in terms of MW distribution of organics that are present. Maximum rejections obtained were 94%, 89% and 72% for OD, color and COD, respectively, when 0.8 microm microfiltration membrane and 400 Da NF membrane were used in series. It was also observed that addition of intermediate UF steps did not increase overall rejections and final permeate flux of NF membrane. Based on these observations, an efficient scheme was offered.


Development of a simulation-based assessment to evaluate the clinical competencies of Korean nursing students.
To describe a simulation-based assessment (SBA) to evaluate the clinical competencies of nursing students in children's health and to compare its results with grade point average (GPA), self-efficacy, topic-specific knowledge, and self-reported clinical competency using the Six-D Scale. This cross-sectional, descriptive study recruited nursing students from a children's health clinical practicum. Students were assigned to either an asthma (n=55) or a type 1 diabetes (n=48) care scenario conducted on a high-fidelity simulator. Clinical competencies were assessed using the global rating scale (GRS) and a checklist. Data on 103 students were analyzed. The SBA-GRS indicated that 64.6%-87.3% of students passed. The SBA-GRS showed a statistically significant positive association with the SBA checklist in both the asthma (rho=.763, p<.001) and the type 1 diabetes (rho=.475, p=.001) group. In the asthma group, the SBA-GRS and checklist showed statistically significant associations with GPA (rho=.413, p=.002 vs. r=.508, p<.001) and the Six-D Scale (rho=.266, p=.049 vs. r=.352, p=.008); in the diabetes group, only the SBA checklist showed a statistically significant association with self-efficacy (r=.339, p=.018) and the Six-D Scale (r=.373, p=.009). Four groups by SBA-GRS had statistically significant differences in scores on the SBA checklist in both groups (F=25.757, p<.001 in the asthma group; F=4.790, p=.006 in the diabetes group) and GPA only in the asthma groups (F=6.095, p<.001). SBA was found to be feasible for nursing students. The GRS and checklist were reasonably correlated with other evaluation methods of student competency, but correlations were better with easier scenarios.


Ophelia syndrome: Hodgkin lymphoma with limbic encephalitis.
Limbic encephalitis, an immune-mediated encephalitis, results from inflammation in the medial temporal lobes. The paraneoplastic form is rare in pediatric population, and frequently precedes tumor diagnosis. A 9-year-old boy receiving chemotherapy for Hodgkin lymphoma, developed headache, temporal lobe seizures, anxiety, hallucinations, short-term memory loss and autonomic disturbances. Magnetic resonance imaging of brain showed features suggestive of limbic encephalitis. Electro-encephalography showed diffuse slowing with no epileptiform discharges. We diagnosed paraneoplastic form of limbic encephalitis. Treatment with steroids and intravenous immunoglobulin failed, and the child died 4 weeks after onset of symptoms. Limbic encephalitis should be kept as differential diagnosis in a child with sub-acutely evolving neuropsychiatric symptoms.


Incorporation of urea nitrogen into fecal protein and plasma protein amino acids in elderly human volunteers after ingestion of lactic acid bacteria.
Health effects of fermented milks have been associated with the metabolic activity of lactic acid bacteria in the gastrointestinal tract. It has been proposed that an increased excretion of urea nitrogen via microbial protein may decrease the workload on kidneys and liver. Therefore, a study was carried out in healthy elderly human subjects to investigate the incorporation of [15N2]urea nitrogen into plasma and fecal proteins and amino acids. Over a period of 10 d 13 healthy elderly subjects ingested daily a freeze-dried microbial preparation which contained different genera of lactic acid bacteria and is used to produce fermented milk products. One of the strains was originally isolated from stool samples of elderly people from the Caucasus region (Lactobacillus plantarum). No stimulation of fecal protein-nitrogen excretion and no increase in 15N-abundances in fecal protein was measured following the administration of the viable microbial preparation and a [15N2]urea bolus. Tentatively, it was concluded that this may have been caused by the inability of the microbial culture to survive the gastro-intestinal passage and (or) by the absence of additional fermentable carbohydrates in the diet as energy source for bacterial protein synthesis in the large intestine. However, using a highly sensitive GC-C-IRMS method we observed a significant incorporation of 15N into plasma protein amino acids. 15N-Enrichments in single amino acids were found according to their participation in transamination reactions. The slight enrichment of lysine which is not transaminated in mammalian tissues may indicate a microbial synthesis and absorption of bacterial lysine.


Feasibility of peripheral blood stem cell collection from sickle cell trait donors with an intensified G-CSF regimen.
Allogeneic hematopoietic stem cell transplantation (HSCT) is the only curative treatment for SCD and bone marrow from an HLA-matched sibling is currently the standard of care. Haploidentical HSCT from a family donor with a TCR αβ/CD19 depleted graft (T-haplo) is an increasingly successful alternative, which requires the generation of G-CSF stimulated peripheral stem cell (PBSC) from haploidentical relatives. These sickle cell trait (SCT) donors reported to develop SCD-related complications in conditions of severe stress. In this retrospective analysis, we compared the safety and efficacy of PBSC mobilization with a G-CSF intensified mobilization regimen in SCT donors with a conventional G-CSF mobilization regimen in healthy donors. The reported adverse events were similar during intensified G-CSF mobilization, apheresis, and shortly after stem cell apheresis in SCT and control donors. In SCT and control donors, we were able to mobilize high yields of CD34+ stem cells and the harvested CD34+ cell count was comparable with control donors. Peripheral stem cell mobilization using an intensified G-CSF regimen is safe, and well tolerated among SCT donors. SCT donors are a valid alternative for collection of peripheral CD34+ stem cells for T-cell-depleted haploidentical stem cell transplantation.


Wetter and better? Changes in associations between drunkenness and other problem behaviors among Norwegian youth.
The consumption of alcohol and the frequency of intoxication among Norwegian youth increased significantly from 1992 to 2002, and we thus asked what, if anything, can be expected to happen to rates of other problem behaviors when heavy drinking becomes more widespread? Do such aggregate level changes in alcohol use imply that individual level associations between drunkenness and behavioral deviance change as well? School-based surveys of 16- to 19-year-old Norwegians that were carried out in 1992 (n = 4,744, response rate 97%) and 2002 (n = 5,458, response rate 92%). Contrary to rates of drunkenness, the level of theft/pilfering declined from 1992 to 2002, while rates of aggressive behavior and school misconduct remained unchanged. We assumed that heavy-drinking adolescents were relatively less deviant with regard to non-violent problem behaviors when the youth culture was relatively 'wet' (2002) as compared to the period when it was much 'drier' (1992) and revealed that this was the case. However, the link between drunkenness and aggressive behavior was equally strong at both assessments. Differential temporal trends in the associations between drunkenness and various problem behaviors may reflect variations with respect to the etiological significance of alcohol use on such behaviors.


Substance P-like immunoreactivity in the pulp-dentine zone of human molar teeth demonstrated by indirect immunofluorescence.
Freshly-extracted human third molars were fixed in Zamboni fixative, demineralized with the mixture of EDTA and the fixative; substance P-like immunoreactivity (SPLI) was revealed by the indirect immunofluorescence technique of Coons. Substance P (SP) was observed in the pulp-dentine zone and the dental pulp. Some of SP-containing fibres ended at the odontoblast layer and did not reach the predentine; others terminated at the predentine surface or penetrated into the predentine. In the predentine, some of SP fibres accompanied odontoblast processes and ended near the mineralized dentine; others changed course transversely at various levels.


Tracking Patient Encounters and Clinical Skills to Determine Competency in Ambulatory Care Advanced Pharmacy Practice Experiences.
To determine if the amount of exposure to patient encounters and clinical skills correlates to student clinical competency on ambulatory care advanced pharmacy practice experiences (APPEs). Students in ambulatory care APPEs tracked the number of patients encountered by medical condition and the number of patient care skills performed. At the end of the APPE, preceptors evaluated students' competency for each medical condition and skill, referencing the Dreyfus model for skill acquisition. Data was collected from September 2012 through August 2014. Forty-six responses from a student tracking tool were matched to preceptor ratings. Students rated as competent saw more patients and performed more skills overall. Preceptors noted minimal impact on workload. Increased exposure to patient encounters and skills performed had a positive association with higher Dreyfus stage, which may represent a starting point in the conversation for more thoughtful design of ambulatory care APPEs.


The chromosomal risk in sperm from heterozygous Robertsonian translocation carriers is related to the sperm count and the translocation type.
To study the chromosomal risk in sperm from Robertsonian translocation (RobT) carriers as a function of the sperm count and translocation type. Prospective study. Departments of reproductive biology, cytogenetics, gynecology, and obstetrics. A total of 29 RobT patients (8 normozoospermic and 21 oligozoospermic) and 20 46,XY patients (10 normozoospermic and 10 oligozoospermic). Sperm fluorescence in situ hybridization with probes for translocation malsegregation and chromosome 13, 18, 21, X, and Y probes for studying the interchromosomal effect (ICE). Translocation malsegregation and ICE aneuploidy rates. In RobT carriers, the sperm translocation malsegregation rate was significantly lower in normozoospermic patients (9.7%) than in oligozoospermic patients (18.0%). Considering only oligozoospermic patients, sperm malsegregation rates were significantly lower for rob(14;21) than for rob(13;14) (11.4% vs. 18.9%). In turn, the rates were significantly lower for rob(13;14) than for rare RobTs (18.9% vs. 25.3%). In sperm from normozoospermic RobT, an ICE was suggested by higher chromosome 13 and 21 aneuploidy rates than in control sperm. Conversely, chromosome 13 and 21 sperm aneuploidy rates were lower in oligozoospermic RobT patients than in oligozoospermic 46,XY patients, but higher than in control subjects. Both translocation type and sperm count influence the RobT malsegregation risk. Of the chromosomes analyzed (13, 18, 21, X, and Y), only chromosomes 13 and 21 were found to be associated with an ICE. Relative to the RobT effect, idiopathic alterations in spermatogenesis in 46,XY patients appear to be more harmful for meiosis.


Intracellularly survived Staphylococcus aureus after phagocytosis are more virulent in inducing cytotoxicity in fresh murine peritoneal macrophages utilizing TLR-2 as a possible target.
Staphylococcus aureus with high virulence potential is contributing to a current public health crisis in both hospital and community settings. TLR-2 and generation of reactive oxygen species (ROS) by phagocytic cells is thought to be an important component of the host's immunity against S. aureus infection. However, response of S. aureus against modulation of host-derived ROS in absence of TLR-2 during acute staphylococcal infection is still remains unclear. Peritoneal macrophages were pretreated with either inhibitors of superoxide dismutase (SOD) or catalase in presence or absence of anti TLR-2 antibody and were infected with S. aureus strain AG-789. Bacteria were recovered after time dependent phagocytosis; intracellular killing, level and expression of SOD and catalase were measured. Phagocytosed bacteria from respective groups were further used for infection to fresh peritoneal macrophages as well as for in vivo infection. Levels of ROS, cytokine, lysozyme, antioxidant enzymes activity and TLR-2 expression were measured. Results revealed that more bacteria were escaped killing in SOD and catalase inhibitor pretreated TLR-2 neutralized macrophages, found to express more catalase and are antibiotic resistant. Infection of fresh macrophages with S. aureus, recovered from SOD and catalase inhibited TLR-2 neutralized macrophages induced lower ROS, lysozyme and cytokine production and caused increased bacterial count. Furthermore, bacterial antioxidants by modulating host-derived ROS could regulate the cell surface TLR-2 expression in murine peritoneal macrophages. So, in the early phase of infection, TLR-2 participates in the innate immune response and targeting bacterial antioxidants might be useful in the alleviation of Staphylococcus aureus infection.


Evaluating the risk of dependence in activities of daily living among community-living older adults with mild to moderate cognitive impairment.
Although cognitive impairment has been implicated as a risk factor for dependence in activities of daily living (ADLs), little is known about the risk of ADL dependence among older adults with cognitive impairment. Among a representative cohort of 1,103 community-living adults aged 72 years and older, we evaluated the 237 subjects with mild to moderate cognitive impairment who were independent in their basic ADLs. All cohort members had undergone a comprehensive assessment, including a battery of qualitative and timed performance tests. ADL dependence developed in 31 (16%) of the 197 subjects who had complete data at the 1-year follow-up. Of the self-reported characteristics, only three--living alone, not being currently married, and having an impairment in IADL function--were significantly associated (p < .05) with the onset of ADL dependence. Several simple tests of physical performance, on the other hand, were strongly associated with the development of ADL dependence. Both timed and qualitative performance tests successfully identified subjects at risk for ADL dependence. A combination of two performance tests--rapid gait and qualitative chair stands--was particularly effective at distinguishing subjects at low (4.7%) and high (34%-39%) risk for ADL dependence. Among community-living older adults with mild to moderate cognitive impairment, the risk of ADL dependence is high but varies considerably depending on how well and how quickly one can perform simple tasks of everyday function. An assessment strategy based on tests of physical performance may allow clinicians to identify subgroups of cognitively impaired elders at low and high risk for ADL dependence.


Removal of tumors in the III ventricle using the lamina terminalis approach. Three cases of isolated growth of craniopharyngiomas in the III ventricle.
Tumors in the III ventricle were totally removed in three children using a route through the lamina terminalis. The cases are discussed on the basis of computed tomography and intraoperative findings. It seems that tumors 4 x 2 cm in size can be successfully removed via this relatively small opening if the neuroradiological findings and the probable histology (craniopharyngioma) provide secure evidence that the tumor site and growth matrix are located in the frontal and lower portion of the III ventricle. Besides the advantage of requiring no transparenchymal access, this quick axial (orthograde) approach exerts no pressure on the hypothalamus, a complication which cannot always be avoided with the transcallosal route or the route through the foramen of Monro. Furthermore, the immediate location of the tumor behind the usually protruding lamina terminalis permits a rapid operation without exploratory characteristics. The distance between the brain surface and the tumor with this procedure is 0 cm; however, it can be up to 9 cm, depending on the age of the patient, with other approaches.


The potential influence of fruit polyphenols on colonic microflora and human gut health.
The effect of common dietary polyphenols on growth of human gut bacteria and their adhesion to enterocytes was investigated. The influence on the growth of a probiotic (Lactobacillus rhamnosus), a commensal (Escherichia coli) and two pathogenic bacteria (Staphylococcus aureus, Salmonella typhimurium) was determined, together with effects on adhesion of pathogenic and probiotic bacteria to cultured Caco-2 cells. All polyphenols, except rutin, were found to affect the viability of representative gut flora in vitro, at doses likely to be present in the gastrointestinal tract, but to differing degrees. Naringenin and quercetin were the most active with the lowest minimum inhibitory concentrations for all the four bacteria tested. The remaining polyphenols had the most marked effect on the Gram positive enteropathogen S. aureus. Naringenin and phloridzin were the most effective inhibitors of S. typhimurium adherence to Caco-2 enterocytes while phloridzin and rutin enhanced the adherence of the probiotic L. rhamnosus. Polyphenols appear to have potential to alter gut microecology and, by affecting the total number of beneficial microflora in the gut, may confer positive gut health benefits.


Fractional flux rates of Nt-methylhistidine in skin and gastrointestine: the contribution of these tissues to urinary excretion of Nt-methylhistidine in the rat.
1. Fractional flux rates of Ntau-methylhistidine (3-methylhistidine; Me-His) of skin and gastrointestine were measured by administering [methyl-3H]methionine to rats. 2. The results showed that the contribution of these tissues to urinary excretion of Me-His was at least 16-6%. This means that when fractional catabolic rates of myosin and actin were estimated from urinary excretion of Me-His, the part of Me-His derived from skin and gastrointestine should not be neglected.


Perihelion Precessions of Inner Planets in Einstein's Theory and Predicted Values for the Cosmological Constant.
In this paper, we obtain the approximate value of 42.9815 arcsec/century for Mercury's perihelion precession by solving both numerically and analytically the nonlinear ordinary differential equation derived from the geodesic equation in Einstein's Theory of Relativity. We also compare our result with known results, and we illustrate graphically the way Mercury's perihelion moves. The results we obtained are applicable to any body that moves around the Sun. We give predictions about the value of the Cosmological Constant. Simple algebraic formulas allow to estimate perihelion shifts with high accuracy.


Distinct role of TGN-resident clathrin adaptors for Vps21p activation in the TGN-endosome trafficking pathway.
Clathrin-mediated vesicle trafficking plays central roles in post-Golgi transport. In yeast (Saccharomyces cerevisiae), the AP-1 complex and GGA adaptors are predicted to generate distinct transport vesicles at the trans-Golgi network (TGN), and the epsin-related proteins Ent3p and Ent5p (collectively Ent3p/5p) act as accessories for these adaptors. Recently, we showed that vesicle transport from the TGN is crucial for yeast Rab5 (Vps21p)-mediated endosome formation, and that Ent3p/5p are crucial for this process, whereas AP-1 and GGA adaptors are dispensable. However, these observations were incompatible with previous studies showing that these adaptors are required for Ent3p/5p recruitment to the TGN, and thus the overall mechanism responsible for regulation of Vps21p activity remains ambiguous. Here, we investigated the functional relationships between clathrin adaptors in post-Golgi-mediated Vps21p activation. We show that AP-1 disruption in the ent3Δ5Δ mutant impaired transport of the Vps21p guanine nucleotide exchange factor Vps9p transport to the Vps21p compartment and severely reduced Vps21p activity. Additionally, GGA adaptors, the phosphatidylinositol-4-kinase Pik1p and Rab11 GTPases Ypt31p and Ypt32p were found to have partially overlapping functions for recruitment of AP-1 and Ent3p/5p to the TGN. These findings suggest a distinct role of clathrin adaptors for Vps21p activation in the TGN-endosome trafficking pathway.


Analysis of chosen SNVs in GPC5, CD58 and IRF8 genes in multiple sclerosis patients.
Multiple sclerosis (MS) is an autoimmune disease of the central nervous system with a neurodegenerative compound. Heterogenetic background of autoimmunity pathway components has been suggested in the MS pathogenesis. The main aim of our study was to evaluate the association between selected polymorphisms of theCD58, IRF8 and GPC5 genes and treatment effectiveness in a group of relapsing-remitting MS patients. This is the first study of MS patients from Podlaskie Region in the Polish population. The study group comprised 174 relapsing-remitting MS patients diagnosed under 40 years of age. Genotyping was performed using ready to use TaqMan assays. We demonstrate a strong association of the polymorphisms with sex, age of onset and response to the treatment applied. A significant correlation was observed in the presence of allele T of rs10492503 polymorphism inGPC5 gene with sex and age of MS onset. Logistic regression analysis revealed an increased risk of the interaction of rs17445836 in IRF8 gene with male sex and the type of treatment (OR = 3.80, p < 0.05), and a decreased risk in the interaction of female sex with disease progress according to the EDSS scale (OR=-2.33, p < 0.05). The analysis of the correlation between different alleles, genotypes and clinical status confirmed the interaction between the genetic factors of age of onset and response to therapy. The study suggests that genetic variants inGPC5, CD58 and IRF8 genes may be of clinical interest in MS as predictors of age of onset and response to therapy.


Switch Loop Flexibility Affects Substrate Transport of the AcrB Efflux Pump.
The functionally important switch loop of the trimeric multidrug transporter AcrB separates the access and deep drug binding pockets in every protomer. This loop, comprising 11-amino-acid residues, has been shown to be crucial for substrate transport, as drugs have to travel past the loop to reach the deep binding pocket and from there are transported outside the cell via the connected AcrA and TolC channels. It contains four symmetrically arranged glycine residues suggesting that flexibility is a key feature for pump activity. Upon combinatorial substitution of these glycine residues to proline, functional and structural asymmetry was observed. Proline substitutions on the PC1-proximal side completely abolished transport and reduced backbone flexibility of the switch loop, which adopted a conformation restricting the pathway toward the deep binding pocket. Two phenylalanine residues located adjacent to the substitution sensitive glycine residues play a role in blocking the pathway upon rigidification of the loop, since the removal of the phenyl rings from the rigid loop restores drug transport activity.


Infusional interleukin-2 and 5-fluorouracil with subcutaneous interferon-alpha for the treatment of patients with advanced renal cell carcinoma: a southwest oncology group Phase II study.
A Phase II trial was conducted to determine the response rate of patients with advanced renal cell carcinoma to a three-drug combination of 5-fluorouracil (5-FU), interleukin-2 (IL-2), and interferon-alpha-2b (IFN-alpha). A 2-stage accrual plan was used that was designed to determine whether response to this regimen was consistent with a true response rate of >/= 30%. The regimen was comprised of 5 treatment days weekly for 4 weeks every 6 weeks. Each weekly treatment was comprised of 5-FU, 1750 mg/m(2), continuous intravenous (i.v.) infusion over 24 hours followed by IL-2, 6 MIU/m(2)/day, continuous i.v. infusion for 4 days. IFN-alpha, 6 MU/m(2), was given subcutaneously on Days 1, 2, and 5. Thirty-eight patients were entered on study, 3 of whom were ineligible. Among the 35 eligible patients there were 3 confirmed partial responses (PR) and 1 complete response (CR), for an overall response rate of 11% (95% confidence interval, 3-27%). One patient considered as having a PR had minimal evidence of residual disease and was free from disease progression at > 2.5 years of follow-up, as was the patient with CR. Three additional patients not qualified as having a PR were showing signs of response at the time they were removed from protocol, and another patient who was removed from protocol early for management of an infection subsequently responded to the same regimen off protocol. Thirteen patients were considered nonassessable (NASS) for response, many of whom had multiple poor risk features and were unable to complete 1 cycle of treatment. This multicenter study failed to confirm an advantageous overall response rate for this three-drug regimen. However, there were two durable responses and indications of responsiveness not scored as PRs among patients with more favorable risk factor patterns, and many poor risk NASS patients. For these reasons, the response rate reported in the current study may be a conservative reflection of the effectiveness of this regimen.


Effect of different PD-1 inhibitor combination therapies for unresectable intrahepatic cholangiocarcinoma.
Immune checkpoint inhibitor (ICI) combination therapy offers a new option for treatment of unresectable intrahepatic cholangiocarcinoma (uICC). To compare the effect of different anti-PD-1 combination therapies as the first-line treatments for uICC. This study included 318 patients who received chemotherapy alone (Chemo), anti-PD-1 plus chemotherapy (ICI-chemo), anti-PD-1 plus targeted therapy (ICI-target) or anti-PD-1 plus targeted therapy and chemotherapy (ICI-target-chemo) as first line for uICC from 22 centres in China. The primary endpoint was progression-free survival (PFS). Secondary endpoints included overall survival (OS), objective response rate (ORR) and safety. Patients with ICI-chemo (median PFS [mPFS], 6.3 months; HR: 0.61, 95% CI: 0.42-0.88; p = 0.008; median OS [mOS], 10.7 months; HR: 0.61, 95% CI: 0.39-0.94; p = 0.026), ICI-target (7.2 months; HR: 0.54, 95% CI: 0.36-0.80; p = 0.002; 15.8 months; HR: 0.54, 95% CI: 0.35-0.84; p = 0.006) or ICI-target-chemo (6.9 months; HR: 0.65, 95% CI: 0.47-0.90; p = 0.009; 14.4 months; HR: 0.47, 95% CI: 0.31-0.70; p < 0.001) achieved better clinical outcomes than those with Chemo (3.8 months; 9.3 months). ICI-target was not inferior to ICI-chemo in survival outcomes (HR for PFS: 0.88, 95% CI: 0.55-1.42; p = 0.614; HR for OS: 0.89, 95% CI: 0.51-1.55; p = 0.680). ICI-target-chemo yielded similar prognoses as ICI-chemo (HR for PFS: 1.07, 95% CI: 0.70-1.62; p = 0.764; HR for OS: 0.77, 95% CI: 0.45-1.31; p = 0.328) and ICI-target (HR for PFS: 1.20, 95% CI: 0.77-1.88; p = 0.413; HR for OS: 0.86, 95% CI: 0.51-1.47; p = 0.583) but resulted in more adverse events (p < 0.001; p = 0.010). Multivariable and propensity score analyses supported these findings. Among patients with uICC, ICI-chemo or ICI-target provided more survival benefits than Chemo while achieving comparable prognoses and fewer adverse events than ICI-target-chemo.


Design of biocomposite materials for bone tissue regeneration.
Several synthetic scaffolds are being developed using polymers, ceramics and their composites to overcome the limitations of auto- and allografts. Polymer-ceramic composites appear to be the most promising bone graft substitute since the natural bone itself is a composite of collagen and hydroxyapatite. Ceramics provide strength and osteoconductivity to the scaffold while polymers impart flexibility and resorbability. Natural polymers have an edge over synthetic polymers because of their biocompatibility and biological recognition property. But, very few natural polymer-ceramic composites are available as commercial products, and those few are predominantly based on type I collagen. Disadvantages of using collagen include allergic reactions and pathogen transmission. The commercial products also lack sufficient mechanical properties. This review summarizes the recent developments of biocomposite materials as bone scaffolds to overcome these drawbacks. Their characteristics, in vitro and in vivo performance are discussed with emphasis on their mechanical properties and ways to improve their performance.


Fear and excitement mark the transition to autonomous, registered nurse.
Gabriella Gilbert, who recently completed her studies to become a fully fledged staff nurse, shares her final reflections on her journey to registration.


Pulmonary delivery alters the disposition of raloxifene in rats.
Pulmonary delivery is an effective way to improve the bioavailability of drugs with extensive metabolism. This research was designed to study the different pharmacokinetic behaviours of small molecule drug after pulmonary delivery and intragastric (i.g) administration. Raloxifene, a selective estrogen receptor modulator with low oral bioavailability (~2%), was chosen as the model drug. Studies were conducted systematically in rats, including plasma pharmacokinetics, excretion, tissue distribution and metabolism. Results showed that raloxifene solution dosed by intratracheal (i.t) administration exhibited relatively quick plasma elimination (t1/2 = 1.78 ± 0.14 h) and undetected absorption process, which was similar with intravenous injection. Compared with i.g administration, the bioavailability increased by 58 times, but the major route of excretion remained faecal excretion. Drug concentration on the bone and the target efficiency were improved by 49.6 times and five times, respectively. Benefited from quick elimination in the lung, chronic toxicity might be ignored. Pulmonary administration improved the bioavailability of raloxifene and further increased the distribution on the target organ (bone), with no obvious impact on its excretory pattern.


[Report of the first case of tinea corporis bullosa by Trichophyton tonsurans].
Blister formation in tinea corporis is rare. Bullous tinea is usually evoked by zoophilic dermatophytes. A 20-year-old woman presented in our out-patient department with a painful pruritic bullous skin eruption at the left forearm. Clinically, a 4 × 3 cm symmetrical plaque with sharp borders and peripheral versiculation and serous crusts was seen. Histologically there was a marked spongiotic dermatitis with fungal elements in periodic acid stain (PAS). By fungal culture, PCR and gene sequencing Trichophyton (T.) tonsurans was identified. To the best of our knowledge, this is the first published case of T. tonsurans as the causative agent of bullous tinea corporis.


Sustained-release isosorbide mononitrate (50 mg): optimization of a once-daily dosage form for long-term treatment of angina pectoris.
A capsule formulation containing 50 mg of isosorbide mononitrate has been developed, 30% of which is in a nonsustained-release form to ensure rapid attainment of initial plasma levels. The remaining 70% is in a sustained-release form that ensures a long-lasting effect and the convenience of once-daily dosing. This formulation of sustained-release isosorbide mononitrate (Elantan LA 50 mg) provides plasma levels greater than 100 ng/ml for 17 hours after oral administration. For the remaining 7 hours, the nitrate plasma level ranges between 50 and 100 ng/ml.


Highly thermostable amylase and pullulanase of the extreme thermophilic eubacterium Rhodothermus marinus: production and partial characterization.
Five strains of the extreme thermophilic Rhodothermus marinus were screened for the production of amylolytic and pullulytic activities. The culture medium for the selected strain, R. marinus ITI 990, was optimized using central composite designs for enhanced enzyme production. The optimized medium containing 1.5 gl(-1) of maltose and 8.3 gl(-1) of yeast extract yielded amylase, pullulanase and alpha-glucosidase activities of 45, 33 and 2.1 nkatml(-1), respectively. Among the various carbon sources tested, maltose was most effective for the formation of these enzymes, followed by soluble maize starch, glycogen and pullulan. The crude amylase and pullulanase showed maximum activities at pH 6.5-7.0, and 85 and 80 degrees C, respectively. At 85 degrees C amylase and pullulanase had half lives of 3 h and 30 min, respectively.


Depression of spinal monosynaptic transmission by diethyl ether: quantal analysis of unitary synaptic potentials.
The site at which diethyl ether impairs transmission in the spinal monosynaptic pathway was studied by intracellular recording from lumbosacral motoneurons. The drug was administered by inhalation to spinal cats, in concentrations which produce surgical anesthesia. Ether had no significant effect on resting potential or input resistance of the cell membrane. It decreased the electrical excitability in some but not all motoneurons. This action may contribute to the depressant effects of ether. Monosynaptic excitatory postsynaptic potentials evoked by impulses in a single Ia afferent fiber (unitary EPSPs) of the triceps surae nerve were recorded from homonymous motoneurons. They were measured and their amplitude distribution was analyzed by a computer-aided procedure. The mean amplitude of the unitary EPSPs was 0.15 to 0.31 mV, and the mean number of transmitter quanta released by each impulse ranged from 1.5 to 4.1 before drug administration. Both values were decreased during inhalation of ether but recovered toward normal after the drug was discontinued. The mean amplitude of the EPSPs produced by one transmitter quantum was 0.08 to 0.15 mV and was not depressed during ether administration. It is concluded that either in anesthetic concentrations depresses synaptic potentials presynaptically by decreasing the amount of excitatory transmitter released, while leaving the chemosensitivity of the postsynaptic membrane unchanged.


Conditional quorum-sensing induction of a cyanide-insensitive terminal oxidase stabilizes cooperating populations of Pseudomonas aeruginosa.
Pseudomonas aeruginosa, an opportunistic pathogen of humans, uses quorum sensing (QS) to regulate the production of extracellular products that can benefit all members of the population. P. aeruginosa can police QS-deficient cheaters by producing hydrogen cyanide, which is also QS regulated; however, the mechanism by which cooperators selectively protect themselves from the toxicity of cyanide remained unresolved. Here, we show that a cyanide-insensitive terminal oxidase encoded by cioAB provides resistance to cyanide, but only in QS-proficient strains. QS-deficient cheaters do not activate cioAB transcription. QS-mediated regulation of cioAB expression depends on production of both cyanide by cooperators (which is QS regulated) and reactive oxygen species (ROS) from cheaters (which is not QS regulated). This type of regulatory system allows cooperating populations to respond, via ROS, to the presence of cheaters, and might allow them to defer the substantial metabolic cost of policing until cheaters are present in the population.


Fluoxetine and vitamin C synergistically inhibits blood-spinal cord barrier disruption and improves functional recovery after spinal cord injury.
Recently we reported that fluoxetine (10 mg/kg) improves functional recovery by attenuating blood spinal cord barrier (BSCB) disruption after spinal cord injury (SCI). Here we investigated whether a low-dose of fluoxetine (1 mg/kg) and vitamin C (100 mg/kg), separately not possessing any protective effect, prevents BSCB disruption and improves functional recovery when combined. After a moderate contusion injury at T9 in rat, a low-dose of fluoxetine and vitamin C, or the combination of both was administered intraperitoneally immediately after SCI and further treated once a day for 14 d. Co-treatment with fluoxetine and vitamin C significantly attenuated BSCB permeability at 1 d after SCI. When only fluoxetine or vitamin C was treated after injury, however, there was no effect on BSCB disruption. Co-treatment with fluoxetine and vitamin C also significantly inhibited the expression and activation of MMP-9 at 8 h and 1 d after injury, respectively, and the infiltration of neutrophils (at 1 d) and macrophages (at 5 d) and the expression of inflammatory mediators (at 2 h, 6 h, 8 h or 24 h after injury) were significantly inhibited by co-treatment with fluoxetine and vitamin C. Furthermore, the combination of fluoxetine and vitamin C attenuated apoptotic cell death at 1 d and 5 d and improved locomotor function at 5 weeks after SCI. These results demonstrate the synergistic effect combination of low-dose fluoxetine and vitamin C on BSCB disruption after SCI and furthermore support the effectiveness of the combination treatment regimen for the management of acute SCI.


Oral fluid for the serological and molecular diagnosis of measles.
Since measles presents mostly in children, a non-invasive sample collection technique such as oral fluid sampling would be very useful in the early detection of measles RNA and antibodies. The aim of this study was to validate the detection of anti-measles IgM and measles virus RNA in oral fluid and to make a comparison with the gold standard methods of ELISA using serum (Enzygnost(®) anti-Measles IgM) and in-house nested reverse transcriptase polymerase chain reaction (RT-PCR) using nasopharyngeal secretions. Three samples each from 73 measles-positive and 44 measles-negative subjects (serum, oral fluid, and nasopharyngeal secretions) were analyzed. The anti-measles IgM ELISA (MicroImmune) on oral fluid was validated against the IgM ELISA (Siemens) for serum and this resulted in a sensitivity of 92% and specificity of 100%. A molecular nested RT-PCR using oral fluid was validated against the standard assay on nasopharyngeal secretions and gave a sensitivity of 100% and specificity of 100%. The results confirm that both serological and molecular oral fluid assays are suitable for routine use. The use of oral fluid samples for the detection of measles virus may encourage patients, general practitioners, and pediatricians to participate in the Belgian measles surveillance system and other epidemiological studies in the framework of the World Health Organization elimination program.


[Nuclear ion channels of the granular neurons from the dentate gyrus].
The nuclear envelope is a part of the endoplasmic reticulum which surrounds the genetic apparatus of the cell. Using the patch-clamp technique we have investigated ion channels in the membranes of nuclei isolated from the neurons of the dentate gyrus. Our research has shown that there is anionic (329 pS) on the outer and anionic (157 pS) and cationic (179 pS) channels on the inner nuclear membrane. Ion channels in the nuclear envelope of the neurons of the dentate gyrus differ much from ones of the neurons of the cerebellum and CA region of the hippocampus. This fact proves that different types of neurons express different sets of ion channels in the nuclear membranes. The physiological role of the ion nuclear channels in the granule cells is not clear but they may be important for the ion balance between the cytoplasm and the lumen of the nuclear envelope as well as endoplasmic reticulum of the cell if the latter represents an adequate model of granule of the some parts of ER.


Automatic and imperative motor activations in stimulus-response compatibility: magnetoencephalographic analysis of upper and lower limbs.
The stimulus-response (S-R) compatibility effect refers to the difference in performance due to the spatial S-R relationship in choice reaction time. We investigated the mechanism of neural activities in S-R compatibility at the level of the primary motor cortices for upper and lower limbs responses using magnetoencephalography (MEG). In the S-R compatible task, subjects were required to respond on the same side of the stimulus light using either an upper or lower limb. In the incompatible task, subjects were required to respond in the reverse manner. Premotor times of upper and lower limbs were faster for the compatible response than for the incompatible response. The neuromagnetic brain activities related to response execution were estimated using a multi-dipole model. Stimulus-locked MEG indicated that the current moments of motor dipoles for both effectors occurred bilaterally and reached the first peak at a constant delay irrespective of whether the task was compatible or incompatible. This indicates that the neural activation of the primary motor cortex is automatically synchronized with the stimulus onset. Response-locked MEG showed that the peak current moment of the motor dipole contralateral to the response was stronger for the compatible task than for the incompatible one regardless of whether the responses were made using the upper or lower limbs. The MEG results suggest that automatic motor activation facilitates imperative motor activation for a compatible response, whereas it is not sufficient to prime imperative motor activation for an incompatible response.


Diagnostic accuracy of high-resolution MR enterography in Crohn's disease: comparison with surgical and pathological specimen.
To determine the accuracy of high-resolution magnetic resonance enterography (HR-MRE) against surgical and histopathology standards in Crohn's disease, and to determine quantitative MRE findings that can differentiate minor from advanced bowel inflammation. Forty-nine consecutive patients who underwent 51 surgical procedures underwent standard MRE and HR-MRE prior to surgery. MRE images were assessed for superficial ulcers, deep ulcers, abscesses, fistulae, and strictures. Quantitative MRE parameters, such as mural thickness, enhancement ratios (ER) of the abnormal bowel, mesentery (ME), and normal bowel (controls), were recorded. MRE findings were compared with surgical and histological results to obtain sensitivity, specificity, and accuracy. Grading of inflammation was compared with MRE parameters for correlation and discriminating power. The sensitivities and specificities of MRE and HR-MRE enterography in the detection of abnormal segments were 0.82 and 0.95 versus 0.86 and 0.95 respectively. HR-MRE was significantly more sensitive than MRE and HR-MRE in the detection of superficial and deep ulcers, fistulae, and abscesses (0.5 versus 0.69; 0.69 versus 0.94; 0.76 versus 0.95; 0.77 versus 1.0, respectively). Mural thickness, ER, and ME had positive correlation with higher grades of inflammation(r = 0.8, 0.66, 0.42, respectively). Regression analysis showed deep and superficial ulcers, ER > 1.85 and mural thickness >4.5 mm were independent predictors of advanced inflammation. HR-MRE has significantly greater diagnostic accuracy as compared to MRE in the diagnosis of bowel ulceration, fistulae and abscesses. Bowel ulcers, mural enhancement ratio >1.85 are strong predictors of advanced inflammation.


Fermented Sipjeondaebo-tang Alleviates Memory Deficits and Loss of Hippocampal Neurogenesis in Scopolamine-induced Amnesia in Mice.
We investigated the anti-amnesic effects of SJ and fermented SJ (FSJ) on scopolamine (SCO)-induced amnesia mouse model. Mice were orally co-treated with SJ or FSJ (125, 250, and 500 mg/kg) and SCO (1 mg/kg), which was injected intraperitoneally for 14 days. SCO decreased the step-through latency and prolonged latency time to find the hidden platform in the passive avoidance test and Morris water maze test, respectively, and both SCO effects were ameliorated by FSJ treatment. FSJ was discovered to promote hippocampal neurogenesis during SCO treatment by increasing proliferation and survival of BrdU-positive cells, immature/mature neurons. In the hippocampus of SCO, oxidative stress and the activity of acetylcholinesterase were elevated, whereas the levels of acetylcholine and choline acetyltransferase were diminished; however, all of these alterations were attenuated by FSJ-treatment. The alterations in brain-derived neurotrophic factor, phosphorylated cAMP response element-binding protein, and phosphorylated Akt that occurred following SCO treatment were protected by FSJ administration. Therefore, our findings are the first to suggest that FSJ may be a promising therapeutic drug for the treatment of amnesia and aging-related or neurodegenerative disease-related memory impairment. Furthermore, the molecular mechanism by which FSJ exerts its effects may involve modulation of the cholinergic system and BDNF/CREB/Akt pathway.


Independent associations of alanine aminotransferase (ALT) levels with cardiovascular risk factor clustering in Chinese adolescents.
To establish the prevalence of elevated serum alanine aminotransferase (ALT) concentrations in Chinese adolescents, and to explore the nature of associations amongst ALT, obesity and cardiovascular risk factors. Anthropometric measurements and fasting plasma glucose, insulin, lipids and ALT were measured in 2102 Chinese adolescents, aged 12-18 years, randomly selected from 14 secondary schools in Hong Kong. The prevalence of elevated ALT levels was 3.2% and 5.9% if abnormal ALT levels were defined as >40 IU/L and >30 IU/L respectively. Using the <25th, 25-75th and >75th percentile values of ALT, all subjects were divided into 3 ALT strata. Using the lowest ALT stratum as referent, the top ALT stratum was associated with obesity and clustering of cardiometabolic-inflammatory risk markers in both genders. After adjusting for age and body mass index (BMI), the highest ALT stratum remained independently associated with diastolic blood pressure and insulin resistance (fasting insulin and Homeostasis Model Assessment, HOMA-IR, HOMA-beta) in boys (p<0.05); and serum triglyceride and HOMA-beta (p=0.008 and 0.014, respectively) in girls. Repeat analysis after excluding subjects with ALT>30 (n=123) or >40 IU/L (n=67) yielded comparable results. In adolescents, top ALT stratum, albeit within normal range, is associated with clustering of cardiovascular risk factors, independent of obesity.


Gastric cancer mesenchymal stem cells derived IL-8 induces PD-L1 expression in gastric cancer cells via STAT3/mTOR-c-Myc signal axis.
The expression of PD-L1 in tumor cells is one of the main causes of tumor immune escape. However, the exact mechanism for regulating PD-L1 expression in gastric cancer (GC) cells remains unclear. Our previous studies have shown that mesenchymal stem cells (MSCs) exert broad immunosuppressive potential, modulating the activity of cells either in innate or adaptive immune system to promote tumor progress. This study aims to investigate whether GCMSCs regulate the PD-L1 expression in GC cells and explore the specific molecular mechanism. The results have shown that GCMSCs enhanced PD-L1 expression in GC cells resulting in the resistance of GC cells to CD8+ T cells cytotoxicity. However, this resistance was attenuated with IL-8 inhibition. Further studies proved that IL-8 derived from GCMSCs induced PD-L1 expression in GC cells via c-Myc regulated by STAT3 and mTOR signaling pathways. Our data indicated that blocking IL-8 derived from GCMSCs may overcome the immune escape induced by PD-L1 in GC cells and provide a potential strategy to enhance the immunotherapy efficiency in GC.


Mosquito host-seeking diel rhythm and chemosensory gene expression is affected by age and Plasmodium stages.
Malaria parasites can affect vector-related behaviours, increasing transmission success. Using Anopheles gambiae and Plasmodium falciparum, we consider the effect of interaction between infection stage and vector age on diel locomotion in response to human odour and the expression of antennal chemosensory genes. We identified age-dependent behavioural diel compartmentalisation by uninfected females post-blood meal. Infection disrupts overall and diel activity patterns compared with age-matched controls. In this study, mosquitoes carrying transmissible sporozoites were more active, shifting activity periods which corresponded with human host availability, in response to human odour. Older, uninfected, blood-fed females displayed reduced activity during their peak host-seeking period in response to human odour. Age- and infection stage-specific changes in odour-mediated locomotion coincide with altered transcript abundance of select chemosensory genes suggesting a possible molecular mechanism regulating the behaviour. We hypothesize that vector-related behaviours of female mosquitoes are altered by infection stage and further modulated by the age post-blood meal of the vector. Findings may have important implications for malaria transmission and disease dynamics.


Pharmacological modulation of lung cancer cells for potassium ion depletion.
Depletion of intracellular potassium ions (K+) is necessary for cells to shrink, induce DNA fragmentation and activate caspases, events which are features of apoptosis. We used 86Rb+ as a K+ analogue to evaluate the possibility of pharmacologically depleting human pulmonary mesothelioma (P31) and small cell lung cancer (U1690) cells of K+, for future use in studies of apoptosis induction. The Na+, K+, 2CI(-)-cotransport inhibitor bumetanide transiently inhibited 86Rb+ influx, but when combined with the Na+, K+, ATPase pump inhibitor ouabain there was a marked and lasting (up to 6 h) 86Rb+ influx inhibition. Cellular K+ efflux was augmented by amphotericin B, digitonin and nigericin. Amphotericin B was an effective 86Rb+ efflux stimulator with low cytotoxicity, whereas digitonin caused cell detachment and nigericin increased LDH release in the U1690 cell line, indicating considerable toxicity of the drugs. It is possible to efficiently reduce intracellular K+ by persistent K+ influx inhibition and simultaneous K+ efflux stimulation with clinically available drugs.


Anticancer activity of N-bis(trifluoromethyl)alkyl-N'-(polychlorophenyl) and N'-(1,2,4-triazolyl) ureas.
A number of N-bis(trifluoromethyl)alkyl-N'-substituted ureas have been synthesized and evaluated for their in vitro anticancer activity against the human cancer cell lines at the National Cancer Institute (NCI, USA). Marked activity was shown for compounds 4a and 5a. The most sensitive cell lines relative to the tested compounds were: 4a UO-31 (renal cancer, logGI(50) -5.62), HS 578T (breast cancer, logGI(50) -5.50), 5a HCC-2998 (colon cancer, logGI(50) -5.94), NCI-H322M (lung cancer, logGI(50) -5.75) and PC-3 (prostate cancer, logGI(50) -5.66).


Pressure support ventilation vs spontaneous ventilation via ProSeal™ laryngeal mask airway in pediatric patients undergoing ambulatory surgery: a randomized controlled trial.
To investigate the advantages of using pressure support ventilation (PSV) vs spontaneous ventilation via ProSeal™ laryngeal mask airway in children undergoing ambulatory surgery. In our ambulatory surgical unit, the use of unassisted spontaneous breathing via laryngeal mask airway is a common anesthetic technique during general anesthesia. However, this may be associated with inadequate ventilation. PSV is a ventilatory mode that is synchronized with the patient's respiratory effort and may improve gaseous exchange under general anesthesia. After the approval from the ethics committee, a randomized controlled trial involving 24 pediatric patients was conducted in our ambulatory surgical unit. They were randomized into two groups, namely Group PSV (receiving PSV) and Group SV (unassisted spontaneous ventilation). Outcome measures included intraoperative respiratory and hemodynamic parameters as well as recovery room data. There were no significant differences in baseline characteristics between the two groups. Patients in Group PSV had lower ETCO(2) (42.8 ± 5.8 vs 50.4 ± 4.0, P = 0.001) and higher expiratory tidal volume per kg bodyweight (8.3 ± 1.8 ml kg(-1) vs 5.8 ± 0.8 ml kg(-1), P = 0.001) compared with patients in Group SV. There were no significant differences in other respiratory and hemodynamic parameters or recovery room data between the two groups. Pressure support ventilation via ProSeal™ laryngeal mask airway during general anesthesia improves ventilation in pediatric patients undergoing ambulatory surgery. However, this did not translate to a difference in clinical outcome among our study patients.


p53, p63 and p73--solos, alliances and feuds among family members.
p53 controls crucial stress responses that play a major role in preventing malignant transformation. Hence, inactivation of p53 is the single most common genetic defect in human cancer. With the recent discovery of two close structural homologs, p63 en p73, we are getting a broader view of a fascinating gene family that links developmental biology with tumor biology. While unique roles are apparent for each of these genes, intimate biochemical cross-talk among family members suggests a functional network that might influence many different aspects of individual gene action. The most interesting part of this family network derives from the fact that the p63 and p73 genes are based on the "two-genes-in-one" idea, encoding both agonist and antagonist in the same open reading frame. In this review, we attempt to present an overview of the current status of this fast moving field.


Cardiac transplantation for congenitally corrected transposition of the great arteries: a case report.
Orthotopic heart transplantation (HTx) was successfully performed in a 46-year-old man with congenitally corrected transposition of the great arteries. Because of the need for an extraordinary length of aorta, we preserved the aortic arch during harvesting of the donor organ. The allograft was implanted using a biatrial technique, but the heart might have to be rotated clockwise due to the posteriorlyy located pulmonary artery. The patient was well at 4 years after HTx. HTx in corrected transposition of the great arteries is technically feasible. Preservation of the aortic arch of the donor's heart may be necessary to achieve a good anatomic correction. The long-term results of such an operation are as good as other HTx procedures.


Penicillin: the medicine with the greatest impact on therapeutic outcomes.
The principal point of this paper is that the discovery of penicillin and the development of the supporting technologies in microbiology and chemical engineering leading to its commercial scale production represent it as the medicine with the greatest impact on therapeutic outcomes. Our nomination of penicillin for the top therapeutic molecule rests on two lines of evidence concerning the impact of this event: (1) the magnitude of the therapeutic outcomes resulting from the clinical application of penicillin and the subsequent widespread use of antibiotics and (2) the technologies developed for production of penicillin, including both microbial strain selection and improvement plus chemical engineering methods responsible for successful submerged fermentation production. These became the basis for production of all subsequent antibiotics in use today. These same technologies became the model for the development and production of new types of bioproducts (i.e., anticancer agents, monoclonal antibodies, and industrial enzymes). The clinical impact of penicillin was large and immediate. By ushering in the widespread clinical use of antibiotics, penicillin was responsible for enabling the control of many infectious diseases that had previously burdened mankind, with subsequent impact on global population demographics. Moreover, the large cumulative public effect of the many new antibiotics and new bioproducts that were developed and commercialized on the basis of the science and technology after penicillin demonstrates that penicillin had the greatest therapeutic impact event of all times.


Common volatiles are major attractants for neonate larvae of the specialist flea beetle Altica koreana (Coleoptera: Chrysomelidae).
Olfactory stimuli play an important role in the host searching of larval phytophagous insects. Previous studies indicate that larvae that have to find feeding sites after hatching are generally attracted to host volatiles. However, there are few studies on the olfactory responses of neonate larvae to host volatiles in cases when those larvae hatched on the host plant. In the present study, we determined the olfactory responses of neonate larvae of the specialist flea beetle, Altica koreana Ogloblin, to host and six non-host plants, using a static-air "arena." Larvae responded significantly to the host plant Potentilla chinensis Ser. and five of six non-host plants, compared to the control. Larvae did not prefer the host plant over the non-host plants (except Artemisia sp.) when offered a choice. Additionally, odours of a non-host plant, which were unattractive to neonate larvae, may have masked the attractive odour of the host plant. These results indicate that common volatiles can play a major role in attracting larvae of this specialist to plants, but attraction to such odours may not be the major mechanism of host choice.


Cultural adaptation and validation of the Family Nursing Practice Scale (FNPS) for use with Spanish-speaking Nursing Degree students.
The task proposed was to conduct a cultural adaptation of the Family Nursing Practice Scale and to assess the psychometric properties of the resulting instrument. Nursing students must obtain sufficient personal competence and confidence to act with patients and their families. For this purpose, an assessment scale is needed to inform teachers of the student's progress and to determine whether further training or changes in teaching methods are required. A cross-sectional study design was used. The researchers conducted this study with 202 students of nursing at two Spanish universities. In the cultural adaptation, the following steps were followed: definition of concepts, translation, back translation, expert group review and implementation by a pilot group with a subsequent cognitive interview. Internal consistency was determined by Cronbach's alpha. Reliability was verified by an initial application of the scale, followed by a repetition after seven days, analysing the results obtained in terms of the intraclass correlation coefficient, the construct validity (by Spearman's non-parametric correlation test) and confirmatory factor analysis (using JASP 0.16.4 statistical software). Cronbach's alpha resulted in 0.95. Spearman's correlation coefficient Family Nursing Practice Scale total with the course year was -0.26 < 0.001. The intraclass correlation coefficient for the total score of the scale was 0.91. The confirmatory factor analysis was performed on a model in which the total score for the scale was considered in relation to the two subscales, and each subscale in relation to its constituent items. The p-value associated with the chi-square was 0.550. The root mean square error of approximation and comparative fit indices presented values of <0.001 (90 % C.I. <0.001-0.048) and 1.000, respectively. These results indicate that the Spanish version presents good internal consistency, construct validity and reliability. Moreover, the confirmatory factor analysis confirms that it presents a good fit to the model initially proposed for this scale for Nursing Degree students.