Production of High Potency Anti-Teschen Disease Serum From Rabbits and Guinea Pigs.
An antiserum of high antibody content against Teschen disease Konratice virus was obtained by inoculating rabbits and guinea pigs with an antigen composed of aluminum gel and virus propagated in swine kidney cell cultures. The rabbit and guinea pig serums neutralized 1,500 TCID(50) of virus at dilutions of 1:5,120 and 1:2,048, respectively. The antibody level in the rabbit serum was tenfold greater than that in convalescent swine serum. Rabbit serum neutralized 2.8 x 10(6) plaque-forming units of the Konratice virus. At a dilution of 1:5,120, this serum neutralized 1,500 TCID(50) of the Reporyje virus. The methods used to prepare and assay the serum are described.


Mechanism of parenchymal enhancement of the liver with a microbubble-based US contrast medium: an intravital microscopy study in rats.
To investigate the mechanism of prolonged contrast material enhancement of the liver observed with the lipid-shell ultrasonographic (US) contrast agent AF0150, with use of intravital microscopy. Eight Sprague-Dawley rats were used. Six received fluorescent microspheres to label the Kupffer cells; two were used as controls. The edge of the middle lobe of the liver was transilluminated with white light. Fluorescent microspheres were observed under fluorescence light. After injection of AF0150, behavior of microbubbles was observed for 6 minutes while viewing a single high-power field. Multiple other fields were then assessed for stationary bubbles and their relation to Kupffer cells. The number of bubbles in motion, aggregated, stationary, and associated with labeled cells were counted. Of 590 bubbles, 34 (5.8%) became stationary and 556 (94.2%) kept moving. Of the 34 stationary microbubbles, 21 dislodged within 30 seconds. Microbubbles were homogeneously distributed throughout the lobule, in contrast to the dominant periportal distribution of the labeled Kupffer cells. Among 83 stationary bubbles observed from all fields of view, only 14 (17%) were associated with fluorescent-labeled cells. The late parenchymal liver enhancement effect of AF0150 is likely not related to Kupffer-cell uptake, but rather to a mechanical slowdown within the sinusoids.


Paternal THC exposure in rats causes long-lasting neurobehavioral effects in the offspring.
Developmental neurotoxicity of a wide variety of toxicants mediated via maternal exposure during gestation is very well established. In contrast, the impacts of paternal toxicant exposure on offspring neurobehavioral function are much less well studied. A vector for paternal toxicant exposure on development of his offspring has been identified. Sperm DNA can be imprinted by chemical exposures of the father. Most but not all of the epigenetic marks in sperm are reprogrammed after fertilization. The persisting epigenetic marks can lead to abnormal genetic expression in the offspring. We have found that paternal delta-9-tetrohydrocannabinol (THC) exposure in rats causes changes in methylation of sperm (Murphy et al., 2018). This is similar to cannabis-associated changes in sperm DNA methylation we found in human males who smoke cannabis (Murphy et al., 2018). In the current study we investigated the intergeneration effects of THC exposure of young adult male rats (0 or 2 mg/kg/day orally for 12 days) to the neurobehavioral development of their offspring. This paternal THC exposure was not found to significantly impact the clinical health of the offspring, including litter size, sex ratio, pup birth weight, survival and growth. However, it did cause a long-lasting significant impairment in attentional performance in the offspring relative to controls when they were tested in adulthood. There was also a significant increase in habituation of locomotor activity in the adult offspring of the males exposed to THC prior to mating. This study shows that premating paternal THC exposure even at a modest dose for a brief period can cause deleterious long-term behavioral effects in the offspring, notably significant impairment in an operant attention task. Further research should be conducted to determine the degree to which this type of risk is seen in humans and to investigate the mechanisms underlying these effects and possible treatments to ameliorate these long-term adverse behavioral consequences of paternal THC exposure.


Biological Activity Evaluation of Some New Benzenesulphonamide Derivatives.
Bacterial resistance to antibiotics has become one of the most challenging problems of infectious disease treatment. Ten new derivatives of benzenesulphonamide bearing carboxamide functionality were synthesized and investigated for their in vivo anti-inflammatory, in vitro anti-microbial and anti-oxidant activities. The base promoted reactions of the appropriate amino acids with substituted benzenesulphonyl chlorides gave the benzene sulphonamides (3a-j) in excellent yields. Palladium mediated amidation of the benzenesulphonamides (3a-j) and butylamine gave the new carboxamides (4a-j) in excellent yield. Compounds 4a and 4c inhibited carrageenan induced rat-paw edema at 94.69, 89.66, and 87.83% each at 1, 2, and 3 h, respectively. In the antimicrobial activity, compound 4d (MIC 6.72 mg/mL) was most potent against E. coli, compound 4h (MIC 6.63 mg/mL) was the most active against S. aureus, compound 4a (MIC 6.67 and 6.45 mg/mL) was most active against P. aeruginosa and S. typhi, respectively, compound 4f (MIC 6.63 mg/mL) was the most active against B. subtilis, compounds 4e and 4h (MIC 6.63 mg/mL) each were the most active against C. albicans, while compound 4e (MIC 6.28 mg/mL) was most active against A. niger. Only compound 4e (IC50 0.3287 mg/mL) had comparable activity with Vitamin C (IC50 0.2090 mg/mL).


Proposal for Improvement of the Hospital Anxiety and Depression Scale for the Assessment of Emotional Distress in Patients With Chronic Musculoskeletal Pain: A Bifactor and Item Response Theory Analysis.
The Hospital Anxiety and Depression Scale (HADS) is a scale originally developed for the assessment of anxiety and depression in hospitalized patients. Despite its wide diffusion, research on its factorial structure has displayed inconsistent results, leaving doubts about its use in chronic musculoskeletal pain. The purpose of this study was to thoroughly assess the factorial structure of the HADS in patients with chronic pain and to give guidance for a potential refinement. Data from 2,522 patients with chronic pain from the Amsterdam Pain (AMS-PAIN) cohort were analyzed through: 1) exploratory bifactor analysis based on a Schmid-Leiman orthogonalization, 2) confirmatory factor analysis comparing a unidimensional model, the original correlated factors model and a bifactor model, 3) item response theory (IRT) analysis based on the graded response model. The results of the confirmatory factor analysis and of the IRT analysis were then cross-validated in an independent sample of patients with chronic pain (n = 8,604). Both exploratory and confirmatory analyses revealed the presence of a strong general emotional distress factor, suggesting that the HADS can be used as a unidimensional scale. The IRT analysis led to the exclusion of three items and to the recoding of one item. The refined 11-item HADS scale was successfully cross-validated and confirmed as a unidimensional, locally independent, monotonic, and reliable scale. PERSPECTIVE: An 11-item shorter version of the HADS could be used to measure emotional distress in patients with chronic musculoskeletal pain. Given its unidimensionality, the use of its total score seems appropriate.


Anteroinferior 2.7-mm versus 3.5-mm plating for AO/OTA type B clavicle fractures: a comparative cohort clinical outcomes study.
To compare the Disability of the Arm, Shoulder, and Hand (DASH) and Constant scores, time to union, rate of union, patient cosmetic satisfaction rate, and the need for secondary procedures between 2.7- and 3.5-mm anteroinferior plating for Arbeitsgemeinschaft für Osteosynthesefragen (AO)/Orthopaedic Trauma Association (OTA) type B clavicle fractures. Retrospective, comparative cohort clinical outcomes study. Level I university trauma center. PATIENTS/PARTICIPATION: Thirty-seven patients with an AO/OTA type B clavicle fracture who underwent open reduction internal fixation with either a 2.7- or 3.5-mm reconstruction plate placed in the anterior-inferior position. The main outcome comparisons included DASH score, Constant score, time to union, rate of union, rate of hardware failure, cosmetic satisfaction, and secondary procedure. DASH score, constant score, time to union, rate of union, cosmetic satisfaction, secondary procedure. At 1-year follow-up, analysis yielded no significant differences in DASH scores (P = 0.26) and Constant Shoulder scores (P = 0.79) between the 2 cohorts. There were no statistically significant differences in the time to union (P = 0.86) and the rate of union (P = 0.49). Although the 2.7-mm cohort had a lower reoperation rate, it was not statistically significant (P = 0.11). However, the 2.7-mm cohort did demonstrate a significantly higher rate of cosmetically acceptable reconstruction (P = 0.003). Compared with 3.5-mm anterior-inferior plating, 2.7-mm anteroinferior plating for AO/OTA type B clavicle fractures leads to significantly higher rates of cosmetic acceptability while reducing the need for a secondary procedure and achieving excellent clinical outcomes as measured by the DASH and Constant scores. There were no differences between the 2.7 and 3.5 cohorts in time to union or in union rate. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


Characterization and verification of CD81 as a potential target in lung squamous cell carcinoma.
CD81 is a cell surface transmembrane protein of the tetraspanin family, which critically regulates signal transduction and immune response. Growing evidence has shown that CD81 plays important roles in tumorigenesis and influences immunotherapy response. Here, combining bio-informatics and functional analysis, we find that CD81 is a risk factor in lung squamous cell carcinoma (LUSC), whereas a protective factor in lung adenocarcinoma. In LUSC with high expression of CD81, the autophagy and JAK-STAT signaling pathway are activated. Meanwhile, the expression level of CD81 is negatively correlated with tumor mutational load (TMB), microsatellite instability (MSI), and neoantigen (NEO). Furthermore, patients with LUSC and high expression of CD81 do not respond to immunotherapy drugs, but can respond to chemotherapy drugs. Importantly, depletion of CD81 suppresses the proliferation of LUSC cell, and enhances the sensitivity to cisplatin. Our findings suggest that CD81 represents a potential target for cisplatin-based chemotherapy in patients with LUSC.


Single cell RT-PCR analysis of ClC-2 mRNA expression in ureteric bud tip.
Embryonic epithelia at the tip of the ureteric bud (UB) face the interspace between epithelial and mesenchymal cells and are fundamentally involved in reciprocal signaling during early nephrogenesis. To characterize their membrane conductive proteins, patch-clamp and single cell RT-PCR techniques were applied to embryonic rat UBs [embryonic day 17 (day E17)] microdissected from the outer cortex. Cells at the UB tip had a high whole cell conductance (14 +/- 2 nS/10 pF, n = 8). The main fractional conductance resembled that of Ca-activated Cl channels in nonepithelial cells, with its time-dependent activation at depolarizing and inactivation at hyperpolarizing voltages. A second Cl-selective current fraction, by contrast, activated slowly during strong hyperpolarization, suggestive of a ClC-2-mediated conductance. To determine the origin of this current, cytoplasm was harvested into the patch pipette, RNA was reverse transcribed, and cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeper gene or the ClC-2 Cl channel was amplified by polymerase chain reaction (PCR). GAPDH and ClC-2 PCR products were identified in 23 and 8 (out of a total of 57) single cell cDNA samples, respectively. ClC-2 PCR products with two different lengths were obtained, which might be due to two alternatively spliced ClC-2 mRNA isoforms. This first and combined approach by patch-clamp and single cell RT-PCR techniques to embryonic epithelia indicates that 1) cells at the UB tip express a phenotype remarkably different from that of postembryonic collecting duct principal cells and that 2) ClC-2 is likely to have a key role in early nephrogenesis.


An open architecture patient monitoring system using standard technologies.
Computer-aided bedside patient monitoring is applied in areas where real-time vital function analysis takes place. Modern bedside monitoring requires not only the networking of bedside monitors with a central monitor but also other standard communication interfaces. In this paper, a novel approach to patient monitoring is introduced. A patient monitoring system was developed and implemented based on an existing industry standard communication network, using standard hardware components and software technologies. The open architecture system design offers scalability, standard interfaces, and flexible signal interpretation possibilities.


Extracellular UTP stimulates electrogenic bicarbonate secretion across CFTR knockout gallbladder epithelium.
The loss of cystic fibrosis transmembrane conductance regulator (CFTR)-mediated transepithelial HCO(3)(-) secretion contributes to the pathogenesis of pancreatic and biliary disease in cystic fibrosis (CF) patients. Recent studies have investigated P2Y(2) nucleotide receptor agonists, e.g., UTP, as a means to bypass the CFTR defect by stimulating Ca(2+)-activated Cl(-) secretion. However, the value of this treatment in facilitating transepithelial HCO(3)(-) secretion is unknown. Gallbladder mucosae from CFTR knockout mice were used to isolate the Ca(2+)-dependent anion conductance during activation of luminal P2Y(2) receptors. In Ussing chamber studies, UTP stimulated a transient peak in short-circuit current (I(sc)) that declined to a stable plateau phase lasting 30-60 min. The plateau I(sc) after UTP was Cl(-) independent, HCO(3)(-) dependent, insensitive to bumetanide, and blocked by luminal DIDS. In pH stat studies, luminal UTP increased both I(sc) and serosal-to-mucosal HCO(3)(-) flux (J(s-->m)) during a 30-min period. Substitution of Cl(-) with gluconate in the luminal bath to inhibit Cl(-)/HCO(3)(-) exchange did not prevent the increase in J(s-->m) and I(sc) during UTP. In contrast, luminal DIDS completely inhibited UTP-stimulated increases in J(s-->m) and I(sc). We conclude that P2Y(2) receptor activation results in a sustained (30-60 min) increase in electrogenic HCO(3)(-) secretion that is mediated via an intracellular Ca(2+)-dependent anion conductance in CF gallbladder.


Tightly regulated and homogeneous transgene expression in human adipose-derived mesenchymal stem cells by lentivirus with tet-off system.
Genetic modification of human adipose tissue-derived multilineage progenitor cells (hADMPCs) is highly valuable for their exploitation in therapeutic applications. Here, we have developed a novel single tet-off lentiviral vector platform. This vector combines (1) a modified tetracycline (tet)-response element composite promoter, (2) a multi-cistronic strategy to express an improved version of the tet-controlled transactivator and the blasticidin resistance gene under the control of a ubiquitous promoter, and (3) acceptor sites for easy recombination cloning of the gene of interest. In the present study, we used the cytomegalovirus (CMV) or the elongation factor 1 α (EF-1α) promoter as the ubiquitous promoter, and EGFP was introduced as the gene of interest. hADMPCs transduced with a lentiviral vector carrying either the CMV promoter or the EF-1α promoter were effectively selected by blasticidin without affecting their stem cell properties, and EGFP expression was strictly regulated by doxycycline (Dox) treatment in these cells. However, the single tet-off lentiviral vector carrying the EF-1α promoter provided more homogenous expression of EGFP in hADMPCs. Intriguingly, differentiated cells from these Dox-responsive cell lines constitutively expressed EGFP only in the absence of Dox. This single tet-off lentiviral vector thus provides an important tool for applied research on hADMPCs.


Novel HIV-1 fusion inhibition peptides: designing the next generation of drugs.
The development of over 20 antiretroviral drugs has led to efficient and successful suppression of HIV-1 replication. In addition to common viral targets, such as reverse transcriptase and protease, new targets have been recently exploited, including integrase, fusion and cellular CCR5. Hence, combination antiretroviral therapy is continually improved by the development of these new agents, especially for patients infected with drug-resistant HIV-1. In this review, we focused on fusion inhibitory peptides that have been developed since the first HIV-1 fusion inhibitor, enfuvirtide (T-20). T-20, approved for clinical use in 2003, is a polypeptide comprising 36 amino acids derived from the HIV-1 gp41 C-terminal heptad repeat and provides a novel treatment strategy for HIV-1 therapy. T-20 is able to suppress HIV-1 replication, including viruses resistant to reverse transcriptase or protease inhibitors. However, after prolonged T-20-containing treatment regimens, HIV-1 acquires resistance to T-20. Therefore, our laboratory and others have developed novel fusion inhibitors, termed next-generation fusion inhibitors, including electrostatically constrained, mutation introduced, and trimer-form peptides.


Predicting Sympathy and Prosocial Behavior from Young Children's Dispositional Sadness.
The purpose of this study was to examine whether dispositional sadness predicted children's prosocial behavior and if sympathy mediated this relation. Constructs were measured when children (N = 256 at Time 1) were 18-, 30-, and 42-months old. Mothers and non-parental caregivers rated children's sadness; mothers, caregivers, and fathers rated children's prosocial behavior; sympathy (concern and hypothesis testing) and prosocial behavior (indirect and direct, as well as verbal at older ages) were assessed with a task in which the experimenter feigned injury. In a panel path analysis, 30-month dispositional sadness predicted marginally higher 42-month sympathy; in addition, 30-month sympathy predicted 42-month sadness. Moreover, when controlling for prior levels of prosocial behavior, 30-month sympathy significantly predicted reported and observed prosocial behavior at 42 months. Sympathy did not mediate the relation between sadness and prosocial behavior (either reported or observed).


Biochemical changes in tissue catecholamines and serotonin in duodenal ulceration caused by cysteamine or propionitrile in the rat.
Previous structure-activity and pharmacologic studies with duodenal ulcerogens cysteamine and propionitrile implicating catecholamines in the pathogenesis of duodenal ulceration have now been followed up by dose- and time-response biochemical investigations to assess the importance of monoamines in the development of duodenal ulcers. The concentrations of norepinephrine (noradrenaline), dopamine, serotonin and their metabolites were measured in total brain, brain regions, stomach, duodenum, pancreas and adrenals in the rat. Turnover of catecholamines was determined in rats pretreated with the inhibitor of tyrosine hydroxylase alpha-methyl-p-tyrosine. The duodenal ulcerogens caused a dose- and time-dependent depletion of norepinephrine in virtually all the tissues examined. The effect was maximal 4 or 7 hr after cysteamine or propionitrile, and norepinephrine levels returned to normal in 24 hr. Dopamine changes were selective and often biphasic, e.g., elevation in adrenals, biphasic in brain cortex, hippocampus and midbrain, but uniformly decreasing in glandular stomach and duodenum. In the median eminence dopamine levels decreased by 181 and 324% at 15 and 30 min, respectively, after cysteamine, but neither dopamine nor 3,4-dihydroxyphenylacetic acid was modified in the periventricular nucleus. Serotonin levels were relatively stable, revealing slight elevations or no changes in most of the tissues. The turnover of norepinephrine was accelerated by both chemicals in virtually all brain regions, but dopamine turnover was affected only in a few areas, e.g., in the corpus striatum and medulla oblongata cysteamine decreased dopamine turnover, whereas propionitrile first (at 1 hr) accelerated then (at 8 hr) significantly suppressed it.(ABSTRACT TRUNCATED AT 250 WORDS)


Tumor-Agnostic Circulating Tumor DNA Testing for Monitoring Muscle-Invasive Bladder Cancer.
Circulating tumor DNA (ctDNA) has recently emerged as a real-time prognostic and predictive biomarker for monitoring cancer patients. Here, we aimed to ascertain whether tumor-agnostic ctDNA testing would be a feasible strategy to monitor disease progression and therapeutic response in muscle-invasive bladder cancer (MIBC) patients after radical cystectomy (RC). Forty-two MIBC patients who underwent RC were prospectively included. Blood samples from these patients were collected at different follow-up time points. Two specific mutations (TERT c.1-124C>T and ATM c.1236-2A>T) were analyzed in the patients' plasma samples by droplet digital PCR to determine their ctDNA status. During a median follow-up of 21 months, 24% of patients progressed in a median of six months. ctDNA status was identified as a prognostic biomarker of tumor progression before RC and 4 and 12 months later (HR 6.774, HR 3.673, and HR 30.865, respectively; p < 0.05). Lastly, dynamic changes in ctDNA status between baseline and four months later were significantly associated with patient outcomes (p = 0.045). In conclusion, longitudinal ctDNA analysis using a tumor-agnostic approach is a potential tool for monitoring MIBC patients after RC. The implementation of this testing in a clinical setting could improve disease management and patients' outcomes.


Flow, packing and compaction properties of novel coprocessed multifunctional directly compressible excipients prepared from tapioca starch and mannitol.
Novel multifunctional excipients were prepared by coprocessing tapioca starch with mannitol using two methods viz; co-grinding and co-fusion. The flow, packing and compaction properties of the native and novel excipients were evaluated by using density, Hausner's ratio, angle of repose, the maximum volume reduction, consolidation index, the rate of consolidation, angle of internal friction, morphological properties, Heckel analysis, tensile strength and dilution potential as evaluation parameters. The study revealed that the method of coprocessing, particle size and particle shape influenced the properties of the resulting novel excipients. Co-grinding was less effective than co-fusion in the preparation of excipients with enhanced properties. The study concluded that coprocessing tapioca starch and mannitol will enhance the flow, packing and compaction properties of the novel excipient and that the co-fusion method of coprocessing would produce novel excipients with enhanced direct compression potential compared to the co-grinding method.


Rapid heteromerization and phosphorylation of ligand-activated plant transmembrane receptors and their associated kinase BAK1.
In plants leucine-rich repeat receptor kinases (LRR-RKs) located at the plasma membrane play a pivotal role in the perception of extracellular signals. For two of these LRR-RKs, the brassinosteroid receptor BRI1 and the flagellin receptor FLS2, interaction with the LRR receptor-like kinase BAK1 (BRI1-associated receptor kinase 1) was shown to be required for signal transduction. Here we report that FLS2.BAK1 heteromerization occurs almost instantaneously after perception of the ligand, the flagellin-derived peptide flg22. Flg22 can induce formation of a stable FLS2.BAK1 complex in microsomal membrane preparations in vitro, and the kinase inhibitor K-252a does not prevent complex formation. A kinase dead version of BAK1 associates with FLS2 in a flg22-dependent manner but does not restore responsiveness to flg22 in cells of bak1 plants, demonstrating that kinase activity of BAK1 is essential for FLS2 signaling. Furthermore, using in vivo phospholabeling, we are able to detect de novo phosphorylation of both FLS2 and BAK1 within 15 s of stimulation with flg22. Similarly, brassinolide induces BAK1 phosphorylation within seconds. Other triggers of plant defense, such as bacterial EF-Tu and the endogenous AtPep1 likewise induce rapid formation of heterocomplexes consisting of de novo phosphorylated BAK1 and proteins representing the ligand-specific binding receptors EF-Tu receptor and Pep1 receptor 1, respectively. Thus, we propose that several LRR-RKs form tight complexes with BAK1 almost instantaneously after ligand binding and that the subsequent phosphorylation events are key initial steps in signal transduction.


Universal platform for sensitive and label-free nuclease assay based on conjugated polymer and DNA/intercalating dye complex.
By taking advantage of the large signal amplification through efficient fluorescence resonance energy transfer (FRET) from the conjugated polymer to the intercalating dye mediated by DNA, a new strategy for nuclease assay has been developed using conjugated polymer and DNA/intercalating dye complex. The discrimination of DNA before or after digestion by nuclease denotes the universal application of the approach, in which either dsDNA or ssDNA substrates could be used for detecting nuclease activity. This method can be extended to most nucleases by simply changing the substrate DNA. The present method is label-free, rapid, and highly sensitive, with a detection limit much better or at least comparable to previous reports. In addition, this assay is easy to implement for visual detection with the assistance of a UV transilluminator. We reason that a similar strategy can be adopted for the detection of other analytes.


Tuberculosis transmission to young children in a South African community: modeling household and community infection risks.
Tuberculosis transmission is determined by contact between infectious and susceptible individuals. A recent study reported a 4% annual risk of child tuberculosis infection in a southern African township. A model was used to explore the interactions between prevalence of adult tuberculosis infection, adult-to-child contacts, and household ventilation, which could result in such a high annual risk of tuberculosis infection. Number of residents per household and tuberculosis incidence were derived from a household census and community tuberculosis registers. Using the Wells-Riley equation and probability analyses of contact between infectious adults with tuberculosis and preschool children, we estimated the annual risk of tuberculosis infection within and outside of the home. There was a mean of 2.2 adults per child-containing household with a 1.35% annual adult smear-positive tuberculosis notification rate. The maximal household annual risk of tuberculosis infection was 3%, which was primarily determined by the number of resident adults. Transmission risk outside the home increased with increasing number of households visited. Transmission probabilities were sensitive to exposure time, ventilation, and period of adult infectivity. The benefits of increased ventilation were greatest when the period of infectivity was reduced. Similar reductions in household transmission could be achieved by increasing ventilation from 2 to 6 air changes/hour or by separating child and adult sleeping areas. The annual risk of tuberculosis infection of preschool children predominantly results from infectious residents in the home. However, even with limited social interactions, a substantial proportion of transmissions may occur from nonresident adults. The benefits of increased ventilation are maximized when the period of infectivity is reduced by prompt treatment of source cases.


Electrolytes in the aging.
The elderly population in the United States continues to grow and is expected to double by 2050. With aging, there are degenerative changes in many organs and the kidney is no exception. After 40 years of age, there is an increase in cortical glomerulosclerosis and a decline in both glomerular filtration rate and renal plasma flow. These changes may be associated with an inability to excrete a concentrated or a dilute urine, ammonium, sodium, or potassium. Hypernatremia and hyponatremia are the most common electrolyte abnormalities found in the elderly and both are associated with a high mortality. Under normal conditions, the elderly are able to maintain water and electrolyte balance, but this may be jeopardized by an illness, a decline in cognitive ability, and with certain medications. Therefore, it is important to be aware of the potential electrolyte abnormalities in the elderly that can arise under these various conditions to prevent adverse outcomes.


[Significance of GABAA receptors for the pathophysiology and therapy of panic disorders].
Gamma aminobutyric acid (GABA) is the most important inhibitory neurotransmitter in the central nervous system. Its action is exerted in the brain through GABA(A) receptors which belong to the family of ligand-gated ion channels. These GABA(A) receptors consist of various subunits and are targets for benzodiazepines, barbiturates, neuroactive steroids, and distinct anticonvulsive agents. Meanwhile, there is considerable evidence that a dysfunction of GABA(A) receptors plays an important role in the pathophysiology of panic disorder. The anxiolytic effects of benzodiazepines are widely used in the treatment of panic disorder. Nevertheless, side effects of benzodiazepines, e.g., dependency and withdrawal symptoms, limit their use as a long-term treatment. In the meantime, antidepressants, especially selective serotonin reuptake inhibitors, comprise first-line treatment in the pharmacotherapy of panic disorder. They interfere with the synthesis of endogenous neuroactive steroids that allosterically modulate GABA(A) receptor function. With regard to experimentally evoked panic attacks in patients with panic disorder and healthy controls, recent investigations demonstrated that enhancing endogenous GABA through the blockade of the GABA transaminase by vigabatrin or inhibition of GABA transporters by tiagabine may exert anxiolytic effects. This novel strategy targeting the GABA binding site of the GABA(A)/benzodiazepine receptor complex and specific agonists for the benzodiazepine binding site present interesting perspectives for the future pharmacotherapy of panic disorder.


Analyses of unintentional intensity modulation in all-fiber acousto-optic tunable filters.
We theoretically and experimentally analyze unintentional intensity modulation phenomena in two types of all-fiber acousto-optic tunable filters utilizing flexural and torsional acoustic waves. Output filter signal at a resonant wavelength shows time-varying oscillations with even- and odd-order harmonics of applied acoustic frequency, which are explained by two factors of static mode coupling and acoustic back reflection. The magnitudes of static coupling and acoustic reflection in our devices are estimated from the measured first and second harmonic modulation powers.


Plastic-Chip-Based Magnetophoretic Immunoassay for Point-of-Care Diagnosis of Tuberculosis.
Tuberculosis (TB) remains a relevant infectious disease in the 21st century, and its extermination is still far from being attained. Due to the extreme infectivity of incipient TB patients, a rapid sensing system for proficient point-of-care (POC) diagnostics is required. In our study, a plastic-chip-based magnetophoretic immunoassay (pcMPI) is introduced using magnetic and gold nanoparticles (NPs) modified with Mycobacterium tuberculosis (MTB) antibodies. This pcMPI offers an ultrasensitive limit of detection (LOD) of 1.8 pg·ml(-1) for the detection of CFP-10, an MTB-secreted antigen, as a potential TB biomarker with high specificity. In addition, by combining the plastic chip with an automated spectrophotometer setup, advantages include ease of operation, rapid time to results (1 h), and cost-effectiveness. Furthermore, the pcMPI results using clinical sputum culture filtrate samples are competitively compared with and integrated with clinical data collected from conventional tools such as the acid-fast bacilli (AFB) test, mycobacteria growth indicator tube (MGIT), polymerase chain reaction (PCR), and physiological results. CFP-10 concentrations were consistently higher in patients diagnosed with MTB infection than those seen in patients infected with nontuberculosis mycobacteria (NTM) (P < 0.05), and this novel test can distinguish MTB and NTM while MGIT cannot. All these results indicate that this pcMPI has the potential to become a new commercial TB diagnostic POC platform in view of its sensitivity, portability, and affordability.


Angiogenesis-targeting microbubbles combined with ultrasound-mediated gene therapy in brain tumors.
The major challenges in gene therapy for brain cancer are poor transgene expression due to the blood-brain barrier (BBB) and neurologic damage caused by conventional intracerebral injection. Non-viral gene delivery using ultrasound-targeted microbubbles (MBs) oscillation via the systematic transvascular route is attractive, but there is currently no high-yielding and targeted gene expression method. In this study, we developed a non-viral and angiogenesis-targeting gene delivery approach for efficient brain tumor gene therapy without brain damage. We developed a VEGFR2-targeted and cationic microbubbles (VCMBs) gene vector for use with transcranial focused ultrasound (FUS) exposure to allow transient gene delivery. The system was tested in a brain tumor model using the firefly luciferase gene and herpes simplex virus type 1 thymidine kinase/ganciclovir (pHSV-TK/GCV) with VCMBs under FUS exposure for transgene expression and anti-tumor effect. In vitro data showed that VCMBs have a high DNA-loading efficiency and high affinity for cancer cells. In vivo data confirmed that this technique enhanced gene delivery into tumor tissues without affecting normal brain tissues. The VCMBs group resulted in higher luciferase expression (3.8 fold) relative to the CMBs group (1.9 fold), and the direct injection group. The tumor volume on day 25 was significantly smaller in rats treated with the pHSV-TK/GCV system using VCMBs under FUS (9.7±5.2mm3) than in the direct injection group (40.1±4.3mm3). We demonstrated the successful use of DNA-loaded VCMBs and FUS for non-viral, non-invasive and targeted gene delivery to brain tumors.


International mobility of hazardous products, industries, and wastes.
The export of hazards to developing countries, frequently associated with the transfer of technology, is an increasing public health problem. It may arise from the export of hazardous products and wastes, or from the transfer of hazardous industries in the absence of appropriate safeguards. Multinational corporations bear a major responsibility for having lower standards of health protection in manufacturing and marketing in the developing countries than in home-country operations. These firms are coming under growing international pressure from concerned citizens, unions, environmental groups, national governments and international organizations, religious groups, the media, and public health professionals.


Alternative Quantifications of Landscape Complementation to Model Gene Flow in Banded Longhorn Beetles [Typocerus v. velutinus (Olivier)].
Rapid progression of human socio-economic activities has altered the structure and function of natural landscapes. Species that rely on multiple, complementary habitat types (i.e., landscape complementation) to complete their life cycle may be especially at risk. However, such landscape complementation has received little attention in the context of landscape connectivity modeling. A previous study on flower longhorn beetles (Cerambycidae: Lepturinae) integrated landscape complementation into a continuous habitat suitability 'surface', which was then used to quantify landscape connectivity between pairs of sampling sites using gradient-surface metrics. This connectivity model was validated with molecular genetic data collected for the banded longhorn beetle (Typocerus v. velutinus) in Indiana, United States. However, this approach has not been compared to alternative models in a landscape genetics context. Here, we used a discrete land use/land cover map to calculate landscape metrics related to landscape complementation based on a patch mosaic model (PMM) as an alternative to the previously published, continuous habitat suitability model (HSM). We evaluated the HSM surface with gradient surface metrics (GSM) and with two resistance-based models (RBM) based on least cost path (LCP) and commute distance (CD), in addition to an isolation-by-distance (IBD) model based on Euclidean distance. We compared the ability of these competing models of connectivity to explain pairwise genetic distances (R ST) previously calculated from ten microsatellite genotypes of 454 beetles collected from 17 sites across Indiana, United States. Model selection with maximum likelihood population effects (MLPE) models found that GSM were most effective at explaining pairwise genetic distances as a proxy for gene flow across the landscape, followed by the landscape metrics calculated from the PMM, whereas the LCP model performed worse than both the CD and the isolation by distance model. We argue that the analysis of a continuous HSM with GSM might perform better because of their combined ability to effectively represent and quantify the continuous degree of landscape complementation (i.e., availability of complementary habitats in vicinity) found at and in-between sites, on which these beetles depend. Our findings may inform future studies that seek to model habitat connectivity in complex heterogeneous landscapes as natural habitats continue to become more fragmented in the Anthropocene.


Measuring management training. Two new instruments.
Seeks to enhance the measurement of training; describes the development of psychometrically sound and practical measures to assess outcomes relating to in-service management training within a National Health Service District. Develops legitimate measurement tools and demonstrates the positive effects of two specific programmes in relation to behavioural change and participant satisfaction. Further work is required to establish how to monitor the effects of the action learning process in terms of cognitive development.


Effect of curcumin on diabetic peripheral neuropathic pain: possible involvement of opioid system.
Neuropathic pain is one of the most common complications of diabetes mellitus. As efficacy and tolerability of current therapy for neuropathic pain are not ideal, we need to develop the novel drug for better treatment. Curcumin as a natural flavonoid from Curcuma longa has considerable effects on nervous system such as, antidepressant, antinociceptive and neuroprotective effects. The present study was designed to investigate the effect of curcumin on diabetic peripheral neuropathic pain and possible involvement of opioid system. A single dose of 60mg/kg streptozotocin was injected intraperitoneally to induce diabetes in rats. STZ-induced diabetic rats were treated with curcumin (50mg/kg/day) acute and chronically. Thermal hyperalgesia and mechanical allodynia were measured on the days 0, 7, 14 and 21 after diabetes induction as behavioral scores of neuropathic pain. Chronic, but not acute, treatment with curcumin prevents the weight loss and attenuates mechanical allodynia in STZ-induced diabetic rats. Pretreatment with naloxone (1mg/kg) significantly reduced anti-allodynic effect of chronic curcumin in von Frey filament test. Our results suggest that curcumin can be considered as a new therapeutic potential for the treatment of diabetic neuropathic pain and the activation of opioid system may be involved in the antinociceptive effect of curcumin.


Adipose Stroma Accelerates the Invasion and Escape of Human Breast Cancer Cells from an Engineered Microtumor.
Approximately 20-25% of human breast tumors are found within an adipose, rather than fibrous, stroma. Adipose stroma is associated with an increased risk of lymph node metastasis, but the causal association between adipose stroma and metastatic progression in human breast cancer remains unclear. We used micropatterned type I collagen gels to engineer ~3-mm-long microscale human breast tumors within a stroma that contains adipocytes and adipose-derived stem cells (ASCs) (collectively, "adipose cells"). Invasion and escape of human breast cancer cells into an empty 120-μm-diameter lymphatic-like cavity was used to model interstitial invasion and vascular escape in the presence of adipose cell-derived factors for up to 16 days. We found that adipose cells hasten invasion and escape by 1-2 days and 2-3 days, respectively. These effects were mediated by soluble factors secreted by the adipose cells, and these factors acted directly on tumor cells. Surprisingly, tumor invasion and escape were more strongly induced by ASCs than by adipocytes. This work reveals that both adipocytes and ASCs accelerate the interstitial invasion and escape of human breast cancer cells, and sheds light on the link between adipose stroma and lymphatic metastasis in human breast cancer. The online version contains supplementary material available at 10.1007/s12195-021-00697-6.


Evaluation of anatomically designed flocked rectal swabs for molecular detection of enteric pathogens in children admitted to hospital with severe gastroenteritis in Botswana.
Two-hundred eighty matched bulk stool and anatomically designed flocked rectal swab samples were collected from children admitted to the hospital with acute diarrhea in Botswana. Their parents were asked about the acceptability of the swab collection method compared with bulk stool sampling. All samples underwent identical testing with a validated 15-target (9 bacterial, 3 viral, and 3 parasite) commercial multiplex PCR assay. The flocked swabs had a 12% higher yield for bacterial pathogen targets (241 versus 212; P = 0.003) compared with that of stool samples, as well as similar yields for viral targets (110 versus 113; P = 0.701) and parasite targets (59 versus 65; P = 0.345). One hundred sixty-four of the flocked swab-stool pairs were also tested with separate laboratory-developed bacterial and viral multiplex assays, and the flocked rectal swabs had a performance that was similar to that seen with commercial assay testing. Almost all parents/guardians found the swabs acceptable. Flocked rectal swabs significantly facilitate the molecular diagnosis of diarrheal disease in children.


Mechanism of Electrospray Supercharging for Unfolded Proteins: Solvent-Mediated Stabilization of Protonated Sites During Chain Ejection.
Proteins that are unfolded in solution produce higher charge states during electrospray ionization (ESI) than their natively folded counterparts. Protein charge states can be further increased by the addition of supercharging agents (SCAs) such as sulfolane. The mechanism whereby these supercharged [M + zH] z+ ions are formed under unfolded conditions remains unclear. Here we employed a combination of mass spectrometry (MS), ion mobility spectrometry (IMS), and molecular dynamics (MD) simulations for probing the ESI mechanism under denatured supercharging conditions. ESI of acid-unfolded apo-myoglobin (aMb) in the presence of sulfolane produced charge states around 27+, all the way to fully protonated (33+) aMb. MD simulations of aMb 27+ to 33+ in Rayleigh-charged water/sulfolane droplets culminated in electrostatically driven protein expulsion, consistent with the chain ejection model (CEM). The electrostatically stretched conformations predicted by these simulations were in agreement with IMS experiments. The CEM involves partitioning of mobile H+ between the droplet and the departing protein. Our results imply that supercharging of unfolded proteins is caused by residual sulfolane that stabilizes protonated sites on the protruding chains, thereby promoting H+ retention on the protein. The stabilization of charged sites is due to charge-dipole interactions mediated by the large dipole moment and the low volatility of sulfolane. Support for this mechanism comes from the experimental observation of sulfolane adducts on the most highly charged ions, a phenomenon previously noted by Venter ( J. Am. Soc. Mass Spectrom. 2012, 23, 489-497). The "CEM supercharging model" proposed here for unfolded proteins is distinct from the charge trapping mechanism believed to be operative during native ESI supercharging.


Dietary phenolics against colorectal cancer--From promising preclinical results to poor translation into clinical trials: Pitfalls and future needs.
Colorectal cancer (CRC) remains a major cause of cancer death worldwide. Over 70% of CRC cases are sporadic and related to lifestyle. Epidemiological studies inversely correlate CRC incidence with the intake of fruits and vegetables but not with their phenolic content. Preclinical studies using in vitro (cell lines) and animal models of CRC have reported anticancer effects for dietary phenolics through the regulation of different markers and signaling pathways. Herein, we review and contrast the evidence between preclinical studies and clinical trials (patients with CRC or at risk, familial adenopolyposis or aberrant crypt foci) investigating the protective effects of curcumin, resveratrol, isoflavones, green tea extracts (epigallocatechin gallate), black raspberry powder (anthocyanins and ellagitannins), bilberry extract (anthocyanins), ginger extracts (gingerol derivatives), and pomegranate extracts (ellagitannins and ellagic acid). To date, curcumin is the most promising polyphenol as possible future adjuvant in CRC management. Overall, the clinical evidence of dietary phenolics against CRC is still weak and the amounts needed to exert some effects largely exceed common dietary doses. We discuss here the possible reasons behind the gap between preclinical and clinical research (inconsistence of results, lack of clinical endpoints, etc.), and provide an outlook and a roadmap to approach this topic.


Maximizing generalization of cognitions and memories after traumatic brain injury.
The purpose of this study was to examine the processes by which survivors of brain injury learn cognitive strategies and the extent to which these strategies can be applied in other situations. The Principles of transfer of training, and generalization formed the basis for specific predictions made about the acquisition and transfer of learning by the patients. Brain-injured patients were tested for their memory of learned lists of words using the part-to-whole learning paradigm. In two separate experiments participants learned part-lists, which varied in the degree of their categorical organizational of items. A test of whole-list learning in the first experiment (n = 60) showed that brain-damaged patients learned and applied the organization skill learned for the part-list items. The second experiment (n = 40) showed that brain-damaged patients were poor at spontaneously organizing material to facilitate recall. The practical implications of these findings are discussed within the transfer of learning model.


Conformational preferences induced by cyclization in orbitides: a vibrational CD study.
Orbitides are bioactive head-to-tail natural cyclic peptides from plant species. Their bioactivity is intrinsically related to the main conformations adopted in solution, whose correct characterization represents an important bottleneck for medicinal chemistry applications. To date, NMR spectrosocopy has been the most frequently used technique to assess the secondary structure of orbitides. Despite the amount of structural information commonly available from NMR, its time scale frequently results in a limited conformational ensemble with a single mean structure, which may not represent the bioactive conformation. Additionally, problems with inter-residue NOE/ROE signals can reduce the accuracy and confidence of the 3D assignments. Vibrational circular dichroism (VCD), on the other hand, has been demostrated as a powerful tool to probe the stereostructure of chiral molecules, including peptides and proteins, with enhanced sensitivity to individual conformations in the condensed phase. Herein, we present the first VCD stereochemical investigation of orbitides. By combining IR/VCD experiments in ACN-d3 and ACN-d3/D2O mixtures with DFT calculations in different levels of theory we were able to determine the solution-state conformational behavior, as well as the main structural restraints induced by cyclization, of the seven-residue orbitide pohlianin A and its linear precursor. VCD results indicated inverse γ-turns as the most prevalent structural motif for the linear precursor in partially aqueous solution, while type I and type VI β-turns were induced by the cyclization process, along with some classic γ-turns. In addition to the conformation of the cyclic peptide already described from NMR data, two previously unidentified conformations with distinct secondary structures were found to significantly populate the sample. This conformational discriminatory power of VCD for both linear and cyclic peptides in different solvent systems may lead to more accurate structural characterization of turn-rich peptidic natural products and help the design of conformationally-tailored peptides for more precise structure-activity relationship results.


[Population viability of Alouatta palliata (Primates: Atelidae) and Cebus capucinus (Primates: Cebidae) at Refugio de Vida Silvestre Privado Nogal, Sarapiquí, Heredia, Costa Rica].
Habitat destruction may cause wildlife population fragmentation and is considered an important factor in small population species extinction. As wildlife populations become smaller, threats to their stability and persistence arise as a result of demographic, environmental and genetic stochastic factors. The aim of this work was to study the effects of population fragmentation on the long term viability of Alouatta palliata and Cebus capucinus populations, at Refugio de Vida Silvestre Privado Nogal, Sarapiquí (RVSPN), Heredia. For this we used the computer software VORTEX to run a population viability analysis (PVA) for both species. The input data of the PVA were taken from the demography structure of the RVSPN, literature sources from the species and from PVA related papers. We evaluated two sets of scenarios: small fragmented populations to reflect the population current state, and one larger and continuous population, to reflect the effect of reforestation actions followed by RVSPN to connect forest fragments. Results suggest that both A. palliata and C. capucinus can survive in isolated forest fragments. However, if different factors as inbreeding depression, catastrophes or habitat loss were incorporated to the scenarios, the small fragmented populations become unstable and the risk of extinction increased for both species. Continuous and larger populations were more robust against the threats incorporated in the scenarios when compared to the current situation of smaller and fragmented populations. The best management option for both species would be to continue reforestation efforts in the area to connect forest fragments, with the result of larger and continuous populations of both species. It is important to continue the observation of both species populations, and to promote a habitat management to reduce the negative effects of stochastic environmental events.


Characterization of Enterococci- and ESBL-Producing Escherichia coli Isolated from Milk of Bovides with Mastitis in Egypt.
This study aimed to investigate the prevalence and antimicrobial resistance of enterococci- and ESBL-producing E. coli isolated from milk of bovine mastitis cases in Egypt. Fifty milk samples of dairy animals were collected from localities in the Nile Delta region of Egypt. Isolates were identified using MALDI-TOF MS, and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Seventeen Enterococcus isolates and eight coliform isolates could be cultivated. Vancomycin resistance rate was high in Ent. faecalis. The VITEK 2 system confirmed all E. coli isolates as ESBL-producing. All Ent. faecalis isolates harbored erm(B), tetL and aac-aphD genes. The vanA gene was detected in Ent. faecalis isolate, vanB was found in other Enterococcus, while one isolate of E. casseliflavus exhibited the vanA gene. E. coli isolates exhibited high prevalence of erm(B) and tetL. E. coli isolates were analyzed by DNA microarray analysis. Four isolates were determined by O-serotyping as O8 (n = 1), O86 (n = 2) and O157 (n = 1). H-serotyping resulted in H11, H12, H21 (two isolates each) and one was of H16 type. Different virulence-associated genes were detected in E. coli isolates including lpfA, astA, celB, cmahemL, intI1 and intI2, and the iroN gene was identified by DNA microarray analysis.


Giant somatotrophinoma without acromegalic features: more "quiet" than "silent": case report.
"Silent" somatotrophinomas are very rare, typically large pituitary adenomas that present with mild or no acromegalic features despite positive immunostaining for growth hormone and autonomous (nonglucose-suppressible) growth hormone secretion. Because of this, they may be amenable to treatment with somatostatin analogues. We report a giant somatotrophinoma (7 cm in maximal diameter) in a young woman with 6 years of amenorrhea who had no clinical features of acromegaly despite frankly elevated serum insulin-like growth factor 1 level at the time of diagnosis. Immunohistochemistry revealed focal strong positive staining for growth hormone in only 10% of the surgical specimen. Mutations in exons 8 or 9 of GNAS1, which are present in 40% of somatotrophinomas, were absent in the surgical specimen. The patient's clinical, biochemical, and radiological presentation is described and is compared with previously reported cases of silent somatotrophinomas. Because of the size and visual symptomatology, partial resection was performed via a transcranial approach. External adjuvant beam radiotherapy was used. As insulin-like growth factor 1 levels remained elevated, treatment with somatostatin analogue is being pursued. This case represents one of the largest somatotrophinomas described to date, and it demonstrates that serum insulin-like growth factor 1 should be measured even in the absence of acromegalic features in patients presenting with apparently nonsecreting macroadenomas. In addition to surgery and radiotherapy, somatostatin analogues may play an important role in controlling tumor growth.


Cross-correlation quantification of dyssynchrony: a new method for quantifying the synchrony of contraction and relaxation in the heart.
Quantification of left ventricular dyssynchrony using Doppler tissue imaging may improve selection of patients who will benefit from cardiac resynchronization therapy. Most methods used to quantify dyssynchrony use a time-to-peak analysis, which is quantitatively simplistic and requires manual identification of systole and selection of peak velocities. We developed and tested a new, highly automatable dyssynchrony parameter, cross-correlation delay (XCD), that does not require identification of systole or manual selection of peak systolic velocities. XCD uses all velocity data points from 3 consecutive beats (approximately 420 points). We tested XCD on 11 members of a positive control group (responders to cardiac resynchronization therapy with a >or=15% reduction in left ventricular end-systolic volume) and 12 members of a negative control group (normal 12-lead electrocardiogram and 2-dimensional echocardiogram findings). We compared XCD to septal-to-lateral delay in time-to-peak (SLD), maximum difference in the basal 2- or 4-chamber times to peak (MaxDiff), and SD of the 12 basal and midwall times-to-peak (Ts-SD). XCD and Ts-SD were significantly different between the positive and negative control groups (both P <or= .0001). SLD and MaxDiff demonstrated no difference between the positive and negative control groups. XCD and Ts-SD were superior to SLD and MaxDiff in discriminating between positive and negative control groups (both P < .01 by receiver operating characteristic comparison). XCD, SLD, MaxDiff, and Ts-SD demonstrated dyssynchrony in 0%, 50%, 58%, and 50% of the negative control group, respectively. XCD was the only parameter that decreased after resynchronization in the positive control group (from 160 +/- 88-69 +/- 61 milliseconds, P = .003). XCD is superior to existing parameters at discriminating patients with left ventricular dyssynchrony from those with normal function.


Presyrinx in children with Chiari malformations.
Presyrinx is a reversible state of spinal cord edema caused by alterations in CSF flow dynamics. Only three pediatric cases have been reported previously. We describe the clinical and radiologic features of presyrinx in six pediatric patients. We electronically searched pediatric spine MRI reports generated at our institution from January 1995 to April 2007 for the keyword "presyrinx" and identified six patients with this radiologic diagnosis. We reviewed the neuroimaging studies and medical records for information regarding symptoms, treatment, and outcome. Of six patients identified with presyrinx, four had a Chiari I malformation and two had a Chiari II malformation. The MRI characteristics of the presyrinx included T2 prolongation, mild indistinct T1 prolongation, and cord enlargement without frank cavitation. Cine phase-contrast MRI studies were performed in three patients and showed severely diminished or absent CSF flow at the foramen magnum. Five patients underwent surgical decompression. All three patients with postoperative spine imaging showed restoration of CSF flow and resolution of the presyrinx. Symptoms of chronic or acute myelopathy attributable to the presyrinx were present in two patients. These symptoms resolved postoperatively. Chiari I and II malformations obstructing CSF flow at the craniocervical junction may cause presyrinx in children. Presyrinx should be considered in the differential diagnosis of chronic or acute myelopathy in patients at risk for abnormal CSF flow dynamics.


Recruitment of human cytomegalovirus immediate-early 2 protein onto parental viral genomes in association with ND10 in live-infected cells.
The human cytomegalovirus (HCMV) immediate-early 2 (IE2) transactivator has previously been shown to form intranuclear, dot-like accumulations in association with subnuclear structures known as promyelocytic leukemia protein (PML) nuclear bodies or ND10. We recently observed that IE2 can form dot-like structures even after infection of PML knockdown cells, which lack genuine ND10. To further analyze the determinants of IE2 subnuclear localization, a recombinant HCMV expressing IE2 fused to the enhanced green fluorescent protein was constructed. We infected primary human fibroblasts expressing Sp100 fused to the autofluorescent protein mCherry while performing live-cell imaging experiments. These experiments revealed a very dynamic association of IE2 dots with ND10 structures during the first hours postinfection: juxtaposed structures rapidly fused to precise co-localizations, followed by segregation, and finally, the dispersal of ND10 accumulations. Furthermore, by infecting PML knockdown cells we determined that the number of IE2 accumulations was dependent on the multiplicity of infection. Since time-lapse microscopy in live-infected cells revealed that IE2 foci developed into viral replication compartments, we hypothesized that viral DNA could act as a determinant of IE2 accumulations. Direct evidence that IE2 molecules are associated with viral DNA early after HCMV infection was obtained using fluorescence in situ hybridization. Finally, a DNA-binding-deficient IE2 mutant could no longer be recruited into viral replication centers, suggesting that the association of IE2 with viral DNA is mediated by a direct DNA contact. Thus, we identified viral DNA as an important determinant of IE2 subnuclear localization, which suggests that the formation of a virus-induced nucleoprotein complex and its spatial organization is likely to be critical at the early stages of a lytic infection.


Some aspects of the radiopharmaceutics of 99m-Tc-phytate.
This paper outlines the method of preparation of 99m-Tc-phytate and its use for scanning the reticuloendothelial system. The tissue distribution studies in mice and rats indicate that 99m-Tc-phytate is a safe and effective liver imaging agent. This was confirmed by preliminary studies in human subjects. The stability of the aqueous tin-phytate complex is remarkably good, and it could be successfully used for more than two months to prepare the labelled product.


Quantitative mass spectrometry defines an oxidative hotspot in hemoglobin that is specifically protected by haptoglobin.
The reaction of hemoglobin (Hb) with hydrogen peroxide (H(2)O(2)) results in free radicals generated at the heme iron, followed by radical transfer to the porphyrin/globin. In the present work, we employed isobaric tagging for relative and absolute quantification (iTRAQ) and a LC-MALDI-MS/MS-based proteomic approach to identify the extent of oxidative changes within tetrameric Hb and dimeric Hb-haptoglobin (Hb-Hp) complexes. Extensive oxidative modifications were found to be restricted to peptides containing alphaTyr42, betaTyr145, and betaCys93. The protein region composed of these peptides appears to define an area of oxidative activity within the Hb tetramer that extends across the critical alpha1beta2/alpha2beta1 interface. Extensive oxidative modifications occurring at betaCys93 indicate that this surface amino acid is an important end point for free radical induced protein oxidation within Hb. Conversely when Hp 1-1 or 2-2 was complexed with dissociable Hb, oxidative changes in Hp complexed dimeric Hb were prevented. This protection was not observed in a stabilized tetrameric Hb, which displays a weak binding affinity for Hp. Therefore, dimerization of Hb and Hp binding may interfere with free radical translocation and play an important role in the overall antioxidant mechanism of Hp. Interestingly, the prevention of peroxide induced Hb amino acid oxidation in purified Hb-Hp1-1 and Hb-Hp2-2 was found to be equal, indicating a phenotype independent specificity in the process of oxidative protection. Taken together, these data suggest differences in oxidative modifications resulting from peroxide induced heme emanated free radical distribution in tetrameric compared to Hp1-1/Hp2-2 stabilized dimeric Hb.


[Cholecystectomy through a short transverse incision].
For cosmetic purposes, it is possible to perform an elective cholecystectomy through a transverse incision of no more than 5 cm. Access is obtained lengthwise through the rectus abdominis. Bleeding must be strictly controlled and cholecystectomy must be performed with great care. Before the operation, it is necessary to ensure that there is no evidence of pathology outside the gall-bladder; nevertheless the common bile duct must be checked during the operation. In the last ten years, this approach has been used in 125 patients (4.6% of cholecystectomies) with excellent results.


Detection of three sialylated tumor-associated antigens in malignant serous effusions.
One hundred thirty-seven fluid samples from patients with malignant diseases, including 58 with lung cancer and 36 with gastric carcinoma, were screened for three tumor-associated carbohydrate antigens. The antigens tested were sialylated Lewis (SLEX) antigen, sialylated SSEA-1 (SLX) antigen, and sialylated Lewis (CA19-9) antigen. Thirty-four fluid samples from patients with benign diseases including 20 with tuberculous pleurisy were also tested for these three antigens as a control. SLEX antigen had the highest positivity rate among the three antigens both in the pleural effusions of patients with pulmonary adenocarcinoma and for all types of lung cancer (64.1% and 46.6%, respectively). In cancers of digestive system, the percentage of SLEX positivity was almost same as that for CA19-9, and higher than that for SLX. A combination assay using the three antigens resulted in an increased rate of detection of tumor-associated antigens (68% of 58 lung cancers including 83% of 39 adenocarcinomas, as well as 77% of 63 digestive system cancers including 84% of 36 gastric carcinomas). Gel filtration of pleural effusions or bile using sephacryl S-1000 showed that these three antigens were eluted in the void volume, indicating a molecular weight of greater than 2 X 10(6). Thus, all 3 antigens possibly exist on carrier protein with a high molecular weight. These observations suggest the potential application of these antigens to distinguish between benign and malignant fluid, and to monitor the effects of the treatment of various types of cancer.


[Neuroophthalmic characteristics of patients with transient cerebrovascular disorders].
Results of complex neuroophthalmological examinations of a group of patients with transient disturbances of the cerebral circulation are considered and discussed. On the basis of clinical, rheoencephalographic, electroencephalographic, ophthalmocalibrometric, ophthalmodynamometric, biomicroscopic (of the conjunctiva vessels) and other ophthalmoscopic findings it has been concluded that the transient disturbances of the cerebral circulation can be differentiated both topically and pathogenetically: this may serve as a basis for individualized pathogenetic therapy.


[Current applications of numeric angiography in the study of cervico-cerebral vessels, the aorta, pulmonary vessels and following vascular surgery].
Numeric angiography is an examination of choice for studying the thoraco-abdominal aorta, whatever the underlying pathological condition, and is often the only angiographic investigation useful and necessary for deciding on therapy in cases of aneurysm. In contrast, results are unsuccessful in approximately 15% of cases when neck vessels are studied. Significant lesions nearly always produce sufficient information for the decision as to therapy to be made. Vascular surgery follow up is an excellent indication for numeric angiography, whatever the surgical technique used or the anatomic site involved. Reliable data can be obtained from pulmonary vessels by the use of this method in patients with pulmonary embolism, arteriovenous malformations, and abnormal venous return.


Identifying and quantifying orphan protein sequences in fungi.
For large regions of many proteins, and even entire proteins, no homology to known domains or proteins can be detected. These sequences are often referred to as orphans. Surprisingly, it has been reported that the large number of orphans is sustained in spite of a rapid increase of available genomic sequences. However, it is believed that de novo creation of coding sequences is rare in comparison to mechanisms such as domain shuffling and gene duplication; hence, most sequences should have homologs in other genomes. To investigate this, the sequences of 19 complete fungi genomes were compared. By using the phylogenetic relationship between these genomes, we could identify potentially de novo created orphans in Saccharomyces cerevisiae. We found that only a small fraction, <2%, of the S. cerevisiae proteome is orphan, which confirms that de novo creation of coding sequences is indeed rare. Furthermore, we found it necessary to compare the most closely related species to distinguish between de novo created sequences and rapidly evolving sequences where homologs are present but cannot be detected. Next, the orphan proteins (OPs) and orphan domains (ODs) were characterized. First, it was observed that both OPs and ODs are short. In addition, at least some of the OPs have been shown to be functional in experimental assays, showing that they are not pseudogenes. Furthermore, in contrast to what has been reported before and what is seen for older orphans, S. cerevisiae specific ODs and proteins are not more disordered than other proteins. This might indicate that many of the older, and earlier classified, orphans indeed are fast-evolving sequences. Finally, >90% of the detected ODs are located at the protein termini, which suggests that these orphans could have been created by mutations that have affected the start or stop codons.


[Study evaluating the impact of a combination of inotropic support and heart rate control on prognosis and stabilization rate in patients with decompensated chronic heart failure (LEGION)].
To evaluate the efficacy of ivabradine and levosimendan in patients with acute circulatory decompensation. The study enrolled 41 patients (20 men and 21 women) aged 61 +/- 9 years, admitted for decompensated heart failure (NYHA functional class IV). Ejection fraction averaged 21.6%. In most patients, systolic blood pressure was below 105 mm Hg and heart rate (HR) was 100-115 beats/min. Central hemodynamics was measured using the strip tests. The patients' clinical status was rated applying the scale accepted in Russia. In addition to conventional therapy, Group 1 patients received levosimendan and ivabradine and Group 2 took dopamine after randomization. A significantly more pronounced clinical improvement was noted in Group 1 patients at hours 48 and 72 of determination. There was a significantly more marked HR reduction in Group 1 at 24 and 72 hours of observation. By day 3 of observation, pulmonary wedge pressure was significantly lower in Group 1 than in Group 2. In the same control periods, the increase in coronary perfusion pressure was significantly obvious in Group 1. The drastically increased level of N-terminal fragment of the prohormone brain-type natriuretic peptide was significantly reduced in both groups, but more considerably in Group 1. In patients with decompensated heart failure on conventional therapy, the co-administration of levosimendan and ivabradine is more effective than the use of dopamine.


(+)Lysergic acid diethylamide, but not its nonhallucinogenic congeners, is a potent serotonin 5HT1C receptor agonist.
Activation of central serotonin 5HT2 receptors is believed to be the primary mechanism whereby lysergic acid diethylamide (LSD) and other hallucinogens induce psychoactive effects. This hypothesis is based on extensive radioligand binding and electrophysiological and behavioral studies in laboratory animals. However, the pharmacological profiles of 5HT2 and 5HT1C receptors are similar, making it difficult to distinguish between effects due to activation of one or the other receptor. For this reason, it was of interest to investigate the interaction of LSD with 5HT1C receptors. Agonist-stimulated phosphoinositide hydrolysis in rat choroid plexus was used as a direct measure of 5HT1C receptor activation. (+)LSD potently stimulated phosphoinositide hydrolysis in intact choroid plexus and in cultures of choroid plexus epithelial cells, with EC50 values of 9 and 26 nM, respectively. The effect of (+)LSD in both systems was blocked by 5HT receptor antagonists with an order of activity consistent with interaction at 5HT1C receptors. Neither (+)-2-bromo-LSD nor lisuride, two nonhallucinogenic congeners of LSD, were able to stimulate 5HT1C receptors in cultured cells or intact choroid plexus. In contrast, lisuride, like (+)LSD, is a partial agonist at 5HT2 receptors in cerebral cortex slices and in NIH 3T3 cells transfected with 5HT2 receptor cDNA. The present finding that (+)LSD, but not its nonhallucinogenic congeners, is a 5HT1C receptor agonist suggests a possible role for these receptors in mediating the psychoactive effects of LSD.


Associations between dietary patterns and hypertension among Korean adults: the Korean National Health and Nutrition Examination Survey (2008-2010).
The objective of this study is to identify the dietary patterns associated with the risk of hypertensions among Korean adults using data from the Korean National Health and Nutrition Examination Survey (KNHANES, 2008-2010). This study analyzes data from 11,883 subjects who participated in the health and nutrition survey, aging from 20 to 64 years. We performed factor analysis based on the weekly mean intake frequencies of 36 food groups to identify major dietary patterns. We identified three major dietary patterns in both sexes, namely "traditional", "western" and "dairy and carbohydrate" patterns. Participants in the highest quartile of western pattern scores had significantly higher blood pressure, serum total cholesterol, and triglyceride levels than those in the lowest quartile. Although not statistically significant, a trend (P for trend = 0.0732) toward a positive association between the western dietary pattern and hypertension risk was observed after adjustments for age, sex, education, income, body mass index (BMI), smoking, physical activity, and energy intake. The dairy and carbohydrate pattern was inversely related with BMI and blood pressures and positively associated with serum high-density lipoprotein (HDL)-cholesterol. After adjusting the age, sex, education, income, BMI, smoking, physical activity and energy intake, the dairy and carbohydrate pattern showed inverse associations with hypertension prevalence (OR = 0.64, 95% CI = 0.55-0.75; P for trend < 0.0001). Intakes of fiber, sodium, and antioxidant vitamins were significantly higher in the top quartile for the traditional pattern than in the lowest quartile for the traditional pattern (P for trend < 0.0001). Intakes of fiber (P for trend < 0.0001), calcium (P for trend < 0.0001), retinol (P for trend = 0.0164), vitamin B1 (P for trend = 0.001), vitamin B2 (P for trend < 0.0001), niacin (P for trend = 0.0025), and vitamin C (P for trend < 0.0001) were significantly increased across quartiles for the dairy and carbohydrate pattern whereas sodium (P for trend < 0.0001) intake was decreased for this pattern. In conclusion, the dairy and carbohydrate pattern may be associated with a reduced risk of hypertension whereas the western pattern may be associated with an increased risk of hypertension among Korean adults.


Male phenotype predicts insemination success in guppies.
Theory predicts that mate choice can lead to an increase in female fecundity if the secondary sexual traits used by females to assess male quality covary with the number of sperm transferred during copulation. Where females mate multiply, such a relationship between male attractiveness and ejaculate size may, additionally (or alternatively), serve to augment the effect of indirect selection by biasing paternity in favour of preferred males. In either case, a positive correlation between male attractiveness and the size of ejaculates delivered at copulation is predicted. To date, some of the most convincing (indirect) evidence for this prediction comes from the guppy, a species of fish exhibiting a resource-free mating system in which attractive males tend to have larger sperm reserves. We show that, during solicited copulations, male guppies with preferred phenotypes actually transfer more sperm to females than their less-ornamented counterparts, irrespective of the size of their initial sperm stores. Our results also reveal that, during coercive copulations, the relationship between ejaculate size and the male's phenotype breaks down. This latter result, in conjunction with our finding that mating speed--a factor under the female's control-is a significant predictor of ejaculate size, leads us to speculate that females may exert at least partial control over the number of sperm inseminated during cooperative matings.


Clinical results of anterior odontoid screw fixation for Type II odontoid fractures.
In this study, 31 patients with a diagnosis of Type II odontoid fractures were reported. All patients were treated with anterior transodontoid screw fixation and clinical outcomes were reported. In this study, the retrospective clinical analysis of 31 patients with traumatic type II odontoid fractures who were treated through anterior transodontoid fixation in Neurosurgery Department at VKV American Hospital between 1998 and 2012 was performed. The age, sex, cause of injury, diagnosis time, neurological examination before and after surgery, follow-up period were evaluated. The neurological status of patients was classified according to the Frankel scale. In 4 patients, 2 transodontoid screws were inserted. The mean hospital stay was 3.35 days. Posterior occipito-cervical fusion was done in 1 patient due to the lack of fusion in the first operation. No vascular injury, screw malposition, infection, neurologic deterioration, or complications were observed during the peroperative and postoperative stage. The mean postoperative follow-up period was 36 months after surgery. Radiological imaging of patients were performed at the early and late postoperative stage. We found satisfying fusion rates and better patient comfort during the postoperative period. We think that stabilization and fusion through a transodontoid screw is a minimal invasive method.


The Tyrosine Kinase-Driven Networks of Novel Long Non-coding RNAs and Their Molecular Targets in Myeloproliferative Neoplasms.
Recent research has focused on the mechanisms by which long non-coding RNAs (lncRNAs) modulate diverse cellular processes such as tumorigenesis. However, the functional characteristics of these non-coding elements in the genome are poorly understood at present. In this study, we have explored several mechanisms that involve the novel lncRNA and microRNA (miRNA) axis participating in modulation of drug response and the tumor microenvironment of myeloproliferative neoplasms (MPNs). We identified novel lncRNAs via mRNA sequencing that was applied to leukemic cell lines derived from BCR-ABL1-positive and JAK2-mutant MPNs under treatment with therapeutic tyrosine kinase inhibitors (TKI). The expression and sequence of novel LNC000093 were further validated in both leukemic cells and normal primary and pluripotent cells isolated from human blood, including samples from patients with chronic myelogenous leukemia (CML). Downregulation of LNC000093 was validated in TKI-resistant CML while a converse expression pattern was observed in blood cells isolated from TKI-sensitive CML cases. In addition to BCR-ABL1-positive CML cells, the driver mutation JAK2-V617F-regulated lncRNA BANCR axis was further identified in BCR-ABL1-negative MPNs. Further genome-wide validation using MPN patient specimens identified 23 unique copy number variants including the 7 differentially expressed lncRNAs from our database. The newly identified LNC000093 served as a competitive endogenous RNA for miR-675-5p and reversed the imatinib resistance in CML cells through regulating RUNX1 expression. The extrinsic function of LNC000093 in exosomal H19/miR-675-induced modulation for the microenvironment was also determined with significant effect on VEGF expression.


Discussion of Alfred Alder's preface to The Diary of Vaslav Nijinsky.
In his preface to The Diary of Vaslav Nijinsky, Alfred Adler (1) found his theory of the dynamics of schizophrenia supported in the Diary, (2) alluded to Nijinsky's prepsychotic personality, and (3) briefly touched on the possibility and conditions of recovery. To add to the understanding of Adler's "Preface," this discussion (1) expands his theory of schizophrenia, (2) gives some concrete data of Nijinsky's prepsychotic personality, (3) describes two episodes of recovery subsequent to the "Preface," and (4) introduces an important aspect of Adler's theory, which he had to omit out of consideration for Nijinsky's wife, Romola-namely, her role in her husband's disorder. With the larger theoretical and historical context established. Adler's "Preface" can be appreciated for its predictive validity.


The State of Squamate Genomics: Past, Present, and Future of Genome Research in the Most Speciose Terrestrial Vertebrate Order.
Squamates include more than 11,000 extant species of lizards, snakes, and amphisbaenians, and display a dazzling diversity of phenotypes across their over 200-million-year evolutionary history on Earth. Here, we introduce and define squamates (Order Squamata) and review the history and promise of genomic investigations into the patterns and processes governing squamate evolution, given recent technological advances in DNA sequencing, genome assembly, and evolutionary analysis. We survey the most recently available whole genome assemblies for squamates, including the taxonomic distribution of available squamate genomes, and assess their quality metrics and usefulness for research. We then focus on disagreements in squamate phylogenetic inference, how methods of high-throughput phylogenomics affect these inferences, and demonstrate the promise of whole genomes to settle or sustain persistent phylogenetic arguments for squamates. We review the role transposable elements play in vertebrate evolution, methods of transposable element annotation and analysis, and further demonstrate that through the understanding of the diversity, abundance, and activity of transposable elements in squamate genomes, squamates can be an ideal model for the evolution of genome size and structure in vertebrates. We discuss how squamate genomes can contribute to other areas of biological research such as venom systems, studies of phenotypic evolution, and sex determination. Because they represent more than 30% of the living species of amniote, squamates deserve a genome consortium on par with recent efforts for other amniotes (i.e., mammals and birds) that aim to sequence most of the extant families in a clade.


Relationship between pharmacokinetic parameters of gentamicin and patient characteristics and/or clinical data in patients with solid organ tumours.
Gentamicin monitoring and the selection of the initial dosage are generally based on the relationship between pharmacokinetic parameters of gentamicin (GPP) and patient characteristics and/or clinical data (PC). However, the number of studies about this relationship in cancer patients is limited. Therefore, the main objective of the present study was to evaluate the relationship between GPP and PC in cancer patients and to identify different subgroups within this group of patients with unique relationship models between GPP and PC. A total of 198 cancer patients were included in the study. Firstly, GPP were estimated by the Sawchuk and Zaske regression method. Then, a linear regression analysis was performed to investigate the relationship between GPP and PC, and lastly subgroups with unique models were identified by comparing their regression models. The results revealed that the variable which was the best predictor of the distribution volume of gentamicine was the dosing weight (DW = IBW + (ABW-IBW), ABW being the actual body weight and IBW the ideal body weight). Creatinine clearance (CLCR) measured by a 24-hour urine collection (CLCRu) was the best predictor of gentamicin clearance (CL). When this value is not available, the CLCR estimated by the formula of Crockcroft and Gault (C-G), can be used. When the C-G formula was used, unique models to predict CL from CLCR were identified for patients who were obese, patients who had received high-dose chemotherapy and, for subjects who had never developed aplasia following chemotherapy. Whichever the model used, the results showed that some variability in pharmacokinetic parameters of gentamicin was not explained by the models, especially in some groups of patients.


Protein kinase C-epsilon regulates sphingosine 1-phosphate-mediated migration of human lung endothelial cells through activation of phospholipase D2, protein kinase C-zeta, and Rac1.
The signaling pathways by which sphingosine 1-phosphate (S1P) potently stimulates endothelial cell migration and angiogenesis are not yet fully defined. We, therefore, investigated the role of protein kinase C (PKC) isoforms, phospholipase D (PLD), and Rac in S1P-induced migration of human pulmonary artery endothelial cells (HPAECs). S1P-induced migration was sensitive to S1P(1) small interfering RNA (siRNA) and pertussis toxin, demonstrating coupling of S1P(1) to G(i). Overexpression of dominant negative (dn) PKC-epsilon or -zeta, but not PKC-alpha or -delta, blocked S1P-induced migration. Although S1P activated both PLD1 and PLD2, S1P-induced migration was attenuated by knocking down PLD2 or expressing dnPLD2 but not PLD1. Blocking PKC-epsilon, but not PKC-zeta, activity attenuated S1P-mediated PLD stimulation, demonstrating that PKC-epsilon, but not PKC-zeta, was upstream of PLD. Transfection of HPAECs with dnRac1 or Rac1 siRNA attenuated S1P-induced migration. Furthermore, transfection with PLD2 siRNA, infection of HPAECs with dnPKC-zeta, or treatment with myristoylated PKC-zeta peptide inhibitor abrogated S1P-induced Rac1 activation. These results establish that S1P signals through S1P(1) and G(i) to activate PKC-epsilon and, subsequently, a PLD2-PKC-zeta-Rac1 cascade. Activation of this pathway is necessary to stimulate the migration of lung endothelial cells, a key component of the angiogenic process.


Emotional vitality in family caregivers: content validation of a theoretical framework.
Emotional vitality may play an important role in determining whether informal caregivers are able to successfully adopt and persist in their roles. This study describes a content validation of a conceptual model of emotional vitality in informal caregivers. A secondary content analysis was performed on transcripts of 30 caregivers who were interviewed about their quality of life in relation to assuming the role of informal caregiver for a family member who had experienced a recent stroke. Caregivers discussed changes in their own health, relationships, roles, finances, participation, and mood after assuming the caregiving role. Using a thematic inductive approach, two raters independently coded the presence and frequency of physical, emotional, and social impacts associated with the caregiving role in order to further develop and validate a conceptual model of caregiver emotional vitality. The interviews provided information that affirmed the relevance of four themes relevant to caregiver emotional vitality previously identified: physical health and well-being; mood regulation; sense of control/mastery of new skills; and participation in meaningful activity. An additional theme of support and recognition from others also emerged. Adopting the informal caregiving role results in major impacts to the caregiver's physical, emotional, and social health. Five core domains appear to meaningfully contribute to emotional vitality of caregivers and may influence their ability to persist in this role over time. Many of the factors that influence emotional vitality in caregivers are potentially modifiable. This new model offers new opportunities for rehabilitation specialists and allied health professionals to develop skill-building interventions that may help caregivers successfully adapt and thrive in the caregiving role.


Drosophila MMP2 regulates the matrix molecule faulty attraction (Frac) to promote motor axon targeting in Drosophila.
Matrix metalloproteinases (MMPs) are widely hypothesized to regulate signaling events through processing of extracellular matrix (ECM) molecules. We previously demonstrated that membrane-associated Mmp2 is expressed in exit glia and contributes to motor axon targeting. To identify possible substrates, we undertook a yeast interaction screen for Mmp2-binding proteins and identified the novel ECM protein faulty attraction (Frac). Frac encodes a multidomain extracellular protein rich in epidermal growth factor (EGF) and calcium-binding EGF domains, related to the vertebrate Fibrillin and Fibulin gene families. It is expressed in mesodermal domains flanking Mmp2-positive glia. The juxtaposition of Mmp2 and Frac proteins raises the possibility that Frac is a proteolytic target of Mmp2. Consistent with this hypothesis, levels of full-length Frac are increased in Mmp2 loss-of-function (LOF) and decreased in Mmp2 gain-of-function (GOF) embryos, indicating that Frac cleavage is Mmp2 dependent. To test whether frac is necessary for axon targeting, we characterized guidance in frac LOF mutants. Motor axons in frac LOF embryos are loosely associated and project ectopically, a phenotype essentially equivalent to that of Mmp2 LOF. The phenotypic similarity between enzyme and substrate mutants argues that Mmp2 activates Frac. In addition, Mmp2 overexpression pathfinding phenotypes depend on frac activity, indicating that Mmp2 is genetically upstream of frac. Last, overexpression experiments suggest that Frac is unlikely to have intrinsic signaling activity, raising the possibility that an Mmp2-generated Frac fragment acts as a guidance cue cofactor. Indeed, we present genetic evidence that Frac regulates a non-canonical LIM kinase 1-dependent bone morphogenetic protein signaling pathway in motoneurons necessary for axon pathfinding during embryogenesis.


Retinal vein occlusion and the prevalence of lipoprotein abnormalities.
Ninety-nine patients with retinal vein occlusion (40 with central, 59 with branch vein occlusion) were investigated for the prevalence of associated diseases for comparison with an age-matched control group. There was a significantly increased prevalence of hyperlipidaemia (p less than 0.001) and hypercholesterolaemia (p less than 0.001) in the group with branch retinal vein occlusion and of hyperlipidaemia (p less than 0.001) and hypercholesterolaemia (p less than 0.02) in the group with central retinal vein occlusion compared with controls. An increased prevalence of hypertriglyceridaemia (p less than 0.001) was also found in patients with central vein occlusion. Other associations were noted (hypertension and oestrogen therapy). Increased serum cholesterol concentrations were found in patients with central vein occlusion (p less than 0.05) and branch vein occlusion (p less than 0.001) compared with controls, and similar tendencies were noted for levels of LDL and HDL cholesterol. Patients with central vein occlusion also had raised levels of serum triglyceride (p less than 0.001) and VLDL-triglyceride (p less than 0.05). The occurrence of both central and branch vein occlusions may be associated with an increased prevalence of hyperlipidaemia and hypertension. Increase of serum lipids may contribute to the aetiology of vein occlusion by altering plasma viscosity or affecting platelet function.


Microparticles from human atherosclerotic plaques promote endothelial ICAM-1-dependent monocyte adhesion and transendothelial migration.
Membrane-shed submicron microparticles (MPs) released following cell activation or apoptosis accumulate in atherosclerotic plaques, where they stimulate endothelial proliferation and neovessel formation. The aim of the study was to assess whether or not MPs isolated from human atherosclerotic plaques contribute to increased endothelial adhesion molecules expression and monocyte recruitment. Human umbilical vein and coronary artery endothelial cells were exposed to MPs isolated from endarterectomy specimens (n=62) and characterized by externalized phosphatidylserine. Endothelial exposure to plaque, but not circulating, MPs increased ICAM-1 levels in a concentration-dependant manner (3.4-fold increase) without affecting ICAM-1 mRNA levels. Plaque MPs harbored ICAM-1 and transferred this adhesion molecule to endothelial cell membrane in a phosphatidylserine-dependent manner. MP-borne ICAM-1 was functionally integrated into cell membrane as demonstrated by the increased ERK1/2 phosphorylation following ICAM-1 ligation. Plaque MPs stimulated endothelial monocyte adhesion both in culture and in isolated perfused mouse carotid. This effect was also observed under flow condition and was prevented by anti-LFA-1 and anti-ICAM-1 neutralizing antibodies. MPs isolated from symptomatic plaques were more potent in stimulating monocyte adhesion than MPs from asymptomatic patients. Plaque MPs did not affect the release of interleukin-6, interleukin-8, or MCP-1, nor the expression of VCAM-1 and E-selectin. These results demonstrate that MPs isolated from human atherosclerotic plaques transfer ICAM-1 to endothelial cells to recruit inflammatory cells and suggest that plaque MPs promote atherosclerotic plaque progression.


[The antitumor activity of elemene is associated with apoptosis].
Elemene, isolated from the Chinese medicinal herb Rhizoma Zedoariae was shown to exhibit antitumor activity in vitro and in vivo to human and murine tumor cells. This novel antineoplastic agent has been demonstrated to have substantial clinical activity against various tumors. In this paper, the mechanisms of antitumor activity of elemene are reported. The in vitro effect of elemene on the growth of leukemia cells was evaluated by MTT assay. The IC50 values of elemene for promyelocytic leukemia HL-60 cells and erythroleukemia K562 cells were found to be 27.5 micrograms/ml and 81 micrograms/ml, respectively, while IC50 for peripheral blood leukocytes (PBL) was 254.3 micrograms/ml. The inhibitory effect of elemene on proliferation of HL-60 cells was associated with cell cycle arrest from, S to G2M phase transition and with induction of apoptosis. The apoptosis of tumor cells was confirmed by DNA ladder formation on gel electrophoresis and characteristic ultrastructural alternations. These results indicate that induction of apoptosis contributes to the mechanisms for antitumor activity of elemene.


Loneliness and social adaptation in Brazilian, Canadian, Chinese and Italian children: a multi-national comparative study.
Research on children's loneliness has been conducted mostly in Western, especially North American, cultures. The purpose of the study was to examine relations between loneliness and social adaptation among children and adolescents in four different societies. A total of 2263 children from grade 3 to grade 6, aged 9 to 12 years, in Brazil, Canada, P. R. China, and Southern Italy participated in the study. The participants completed a self-report measure of loneliness. Information about social behaviors and peer relationships was obtained from peer assessments. Multi-group analyses revealed that the overall patterns of relations among social behaviors, peer relationships and loneliness differed across the samples. Specifically, sociability was positively associated with peer relationships and made negative indirect contributions to loneliness through peer relationships in all four samples. Aggression made significant indirect contributions to the prediction of loneliness in Chinese children, but not in other samples. Shyness-sensitivity was associated with loneliness directly in Brazilian and Italian children and indirectly through peer relationships in Canadian children, but not associated with loneliness in Chinese children. The results suggest that the nature of children's loneliness may be affected by the broad socio-cultural context.


The RNA modification database.
The RNA modification database provides a comprehensive listing of post-transcriptionally modified nucleosides from RNA and is maintained as an updated version of the initial printed report. Information provided includes: type(s) of RNA in which found and phylogenetic distribution; common chemical names and symbols; Chemical Abstracts registry numbers and index names; chemical structures; initial literature citations for structural characterization or occurrence and chemical synthesis. The data are available through the World Wide Web, anonymous ftp or from the authors in printed form.


Safety of catheter ablation of atrial fibrillation using fiber optic-based contact force sensing.
While the body of evidence showing the benefit of contact force (CF)-sensing catheters in improving atrial fibrillation ablation success rate is growing, real-world safety data of this technology are limited. The aim of this study was to report the complication rate in patients undergoing ablation using a specific CF-sensing catheter (TactiCath Quartz CF ablation catheter, Abbott, St. Paul, MN) in a large worldwide database. User-reported adverse event data from January 1, 2014 to December 31, 2015 were reviewed for the ablation catheter. These adverse events included cardiac perforation, cerebrovascular accident, atrioesophageal (AE) fistula, steam pop, and death. Rates of these adverse events, including geographic and temporal relationships, were calculated. Adverse events were reported in 178 of 41,709 patients (adverse event rate 0.427%). Adverse events included 117 cardiac perforations (0.281%), 5 cerebrovascular accidents (0.012%), 10 AE fistulas (0.024%), 53 audible steam pops (0.127%), and 13 deaths (0.031%). The total event rate in European countries was 0.53% vs 0.38% in the United States (P = .0082). The AE fistula event rate peaked in late 2014 and early 2015 at 0.06% and decreased over time to a steady-state 0.01%, corresponding to the initial US release, subsequent increased use, and training on the manufacturer recommended settings. Complication rates while using the TactiCath CF ablation catheter appear to decrease with training and experience. The major complication rate associated with this technology is low in the context of similar reports of conventional radiofrequency catheters.