Uncovering Influential Factors in Human Antibiotic Prescribing: A Meta-Synthesis Study Informed by the Theoretical Domains Framework.
This study aimed to identify modifiable determinants (facilitators and barriers) related to the choice of prescribing antibiotics in human medicine across clinical settings. Enhanced management of antibiotics can help slow the spread of resistant bacteria. A qualitative meta-synthesis approach was carried out according to Sandelowski and Barroso's method. Included studies were evaluated using the Critical Appraisal Skills Programme. Findings were extracted and organised to form a qualitative meta-summary. The Theoretical Domains Framework, the Capabilities-Opportunities-Motivation (COM-B) model and the Behaviour Change Weel were used as a coding matrix for data interpretation. The analysis of 63 included studies revealed barriers and facilitators in 12 out of 14 domains specified by the Theoretical Domains Framework. Prescribers' capabilities, motivation and opportunities were found to be the main drivers of antibiotic prescribing behaviour. Knowledge, skills, beliefs, expectations, the influence of patients and colleagues, organisational culture and infrastructure characteristics have a significant impact on prescribing behaviours. A comprehensive inventory of factors related to antibiotic prescribing has been compiled. Interventions to promote appropriate antibiotic prescribing should take a systemic approach rather than focusing solely on individual-level variables. Furthermore, the adoption of co-design approaches for such interventions is desirable to ensure greater applicability and sustainability in the real-world context of organisations.


Changes in Reaction Time after Yoga Bellows-type Breathing in Healthy Female Volunteers.
Previously, yoga bellows-type breathing (bhastrika pranayama) reduced reaction time (RT) or reduced anticipatory responses in male participants or a mixed group of male and female participants. The present study as a control trial aimed to assess the effects of yoga bellows-type breathing on RT in females exclusively. The sample consisted of 25 healthy females, aged between 19 and 32 years (group mean ± standard deviation, 22.8 ± 3.5 years). All of them had prior minimum experience of yoga including yoga bellows-type breathing of 12 months. The RT was assessed in each participant before and after three randomized sessions differed in the intervention given held on three separate days. The sessions were (i) YOGA bellows-type breathing or bhastrika pranayama(BHK), (ii) Breath awareness (BAW), and (iii) Sitting quietly (CTL) as a control session. The duration of the intervention was 18 min, and the participants were assessed for RT before and after the intervention. Repeated measures ANOVA, post hoc tests with Bonferroni adjusted showed that the time taken to obtain a correct response reduced significantly after 18 min of BAW (P < 0.05) and CTL (P < 0.05). However, no changes were seen in the RT after BHK. The results suggest that different interventions may optimize performance in tasks requiring attention in females compared to males.


Hepatic, metabolic and toxic effects of ethanol: 1991 update.
Until two decades ago, dietary deficiencies were considered to be the only reason for alcoholics to develop liver disease. As the overall nutrition of the population improved, more emphasis was placed on secondary malnutrition and direct hepatotoxic effects of ethanol were established. Ethanol is hepatotoxic through redox changes produced by the NADH generated in its oxidation via the alcohol dehydrogenase pathway, which in turn affects the metabolism of lipids, carbohydrates, proteins, and purines. Ethanol is also oxidized in liver microsomes by an ethanol-inducible cytochrome P-450 (P-450IIE1) that contributes to ethanol metabolism and tolerance, and activates xenobiotics to toxic radicals thereby explaining increased vulnerability of the heavy drinker to industrial solvents, anesthetic agents, commonly prescribed drugs, over-the-counter analgesics, chemical carcinogens, and even nutritional factors such as vitamin A. In addition, ethanol depresses hepatic levels of vitamin A, even when administered with diets containing large amounts of the vitamin, reflecting, in part, accelerated microsomal degradation through newly discovered microsomal pathways of retinol metabolism, inducible by either ethanol or drug administration. The hepatic depletion of vitamin A is strikingly exacerbated when ethanol and other drugs were given together, mimicking a common clinical occurrence. Microsomal induction also results in increased production of acetaldehyde. Acetaldehyde, in turn, causes injury through the formation of protein adducts, resulting in antibody production, enzyme inactivation, decreased DNA repair, and alterations in microtubules, plasma membranes and mitochondria with a striking impairment of oxygen utilization. Acetaldehyde also causes glutathione depletion and lipid peroxidation, and stimulates hepatic collagen production by the vitamin A storing cells (lipocytes) and myofibroblasts.(ABSTRACT TRUNCATED AT 250 WORDS)


Ubiquitin/proteasome pathway regulates levels of retinoic acid receptor gamma and retinoid X receptor alpha in human keratinocytes.
Repeated exposure of human skin to solar UV radiation leads to premature aging (photoaging) and skin cancer. UV-induced skin damage can be ameliorated by all-trans retinoic acid treatment. The actions of retinoic acid in skin keratinocytes are mediated primarily by nuclear retinoic acid receptor gamma (RARgamma) and retinoid X receptor alpha (RXRalpha). We found that exposure of cultured primary human keratinocytes to UV irradiation (30 mJ/cm2) substantially reduced (50-90%) RARgamma and RXRalpha mRNA and protein within 8 h. The rates of disappearance of RARgamma and RXRalpha proteins after UV exposure or treatment with the protein synthesis inhibitor cycloheximide were similar. UV irradiation did not increase the rate of breakdown of RARgamma or RXRalpha but rather reduced their rate of synthesis. The addition of proteasome inhibitors MG132 and LLvL, but not the lysosomal inhibitor E64, prevented loss of RARgamma and RXRalpha proteins after exposure of keratinocytes to either UV radiation or cycloheximide. Soluble extracts from nonirradiated or UV-irradiated keratinocytes possessed similar levels of proteasome activity that degraded RARgamma and RXRalpha proteins in vitro. Furthermore, RARgamma and RXRalpha were polyubiquitinated in intact cells. RXRalpha was found to contain two proline, glutamate/aspartate, serine, and threonine (PEST) motifs, which confer rapid turnover of many short-lived regulatory proteins that are degraded by the ubiquitin/proteasome pathway. However, the PEST motifs in RXRalpha did not function to regulate its stability, because deletion of the PEST motifs individually or together did not alter ubiquitination or proteasome-mediated degradation of RXRalpha. These results demonstrate that loss of RARgamma and RXRalpha proteins after UV irradiation results from degradation via the ubiquitin/proteasome pathway. Taken together, the data here indicate that ubiquitin/proteasome-mediated breakdown is an important mechanism regulating the levels of nuclear retinoid receptors.


Identification of a conserved 8 aa insert in the PIP5K protein in the Saccharomycetaceae family of fungi and the molecular dynamics simulations and structural analysis to investigate its potential functional role.
Homologs of the phosphatidylinositol-4-phosphate-5-kinase (PIP5K), which controls a multitude of essential cellular functions, contain a 8 aa insert in a conserved region that is specific for the Saccharomycetaceae family of fungi. Using structures of human PIP4K proteins as templates, structural models were generated of the Saccharomyces cerevisiae and human PIP5K proteins. In the modeled S. cerevisiae PIP5K, the 8 aa insert forms a surface exposed loop, present on the same face of the protein as the activation loop of the kinase domain. Electrostatic potential analysis indicates that the residues from 8 aa conserved loop form a highly positively charged surface patch, which through electrostatic interaction with the anionic portions of phospholipid head groups, is expected to play a role in the membrane interaction of the yeast PIP5K. To unravel this prediction, molecular dynamics (MD) simulations were carried out to examine the binding interaction of PIP5K, either containing or lacking the conserved signature insert, with two different membrane lipid bilayers. The results from MD studies provide insights concerning the mechanistic of interaction of PIP5K with lipid bilayer, and support the contention that the identified 8 aa conserved insert in fungal PIP5K plays an important role in the binding of this protein with membrane surface. Proteins 2017; 85:1454-1467. © 2017 Wiley Periodicals, Inc.


[Role of blood lipoproteins in adaptive reorganization of the rat liver mitochondria].
It was shown in experiments in vitro that high density lipids (HDL), very low density lipids (VLDL) and cAMP taken separately did not affect the kinetics of mitochondrial swelling in the liver of control and 72-hr-fasting rats. ApoHDL and particularly ApoVLDL obtained by delipidation of appropriate lipoproteins had pronounced capacities to stimulate the process of swelling. Mitochondrial swelling increased to an ever greater degree under the action of cAMP coupled with apoproteins. The maximum stimulation of the swelling was attained as a result of administering apoVLDL in conjunction with cAMP. The effects of enhanced swelling were the most remarkable in experiments with mitochondria from the control animals.


Abbreviated course of radiation therapy in older patients with glioblastoma multiforme: a prospective randomized clinical trial.
To prospectively compare standard radiation therapy (RT) with an abbreviated course of RT in older patients with glioblastoma multiforme (GBM). One hundred patients with GBM, age 60 years or older, were randomly assigned after surgery to receive either standard RT (60 Gy in 30 fractions over 6 weeks) or a shorter course of RT (40 Gy in 15 fractions over 3 weeks). The primary end point was overall survival. The secondary end points were proportionate survival at 6 months, health-related quality of life (HRQoL), and corticosteroid requirement. HRQoL was assessed using the Karnofsky performance status (KPS) and Functional Assessment of Cancer Therapy-Brain (FACT-Br). All patients had died at the time of analysis. Overall survival times measured from randomization were similar at 5.1 months for standard RT versus 5.6 months for the shorter course (log-rank test, P =.57). The survival probabilities at 6 months were also similar at 44.7% for standard RT versus 41.7% for the shorter course (lower-bound 95% CI, -13.7). KPS scores varied markedly but were not significantly different between the two groups (Wilcoxon test, P =.63). Low completion rates of the FACT-Br (45%) precluded meaningful comparisons between the two groups. Of patients completing RT as planned, 49% of patients (standard RT) versus 23% required an increase in posttreatment corticosteroid dosage (chi(2) test, P =.02). There is no difference in survival between patients receiving standard RT or short-course RT. In view of the similar KPS scores, decreased increment in corticosteroid requirement, and reduced treatment time, the abbreviated course of RT seems to be a reasonable treatment option for older patients with GBM.


How the Presence of Others Affects Desirability Judgments in Heterosexual and Homosexual Participants.
Mate-choice copying is a mating strategy wherein women rely on contextual information to assist in securing accurate assessments of potential mates. Mate-choice copying has been extensively studied in non-human species and has begun to be examined in humans as well. Hill and Buss (2008) found evidence of opposing effects for men and women in desirability judgments based on the presence of other opposite-sex people. The current study successfully replicated these findings with 73 and 44 heterosexual men and women, respectively. Heterosexual men exhibited the desirability diminution effect, and heterosexual women exhibited the desirability enhancement effect. The current study also extended these findings to include 73 gay men and 32 lesbian women. Findings for gay and lesbian participants were inverted compared to heterosexual participants. Gay men exhibited the desirability enhancement effect, and lesbian women exhibited the desirability diminution effect, revealing sex differences in mate-choice copying spanning different sexual orientations.


Pain, aging and dementia: Towards a biopsychosocial model.
Dementia is a progressive disease associated with irreversible impairment and loss of cognitive abilities. About half of older people with dementia experience pain. In this paper, we propose that pain in older people with dementia can be conceptualized as the final result of the interaction of three heterogeneous phenomena, pain, aging, and dementia, which are created and influenced by the interactions of predisposing, lifelong, and current biopsychosocial factors. We review pain assessment in people with dementia using both self-report and observational/behavioral measures. We then review the biological/sensory, psychological (cognitive and affective) and social dimensions of pain in dementia. The available data suggest that dementia does not impact pain threshold or tolerance. To date, there is little research on the social dimension of pain in dementia. Changes in the affective domain in response to experimental pain have been contradictory with evidence supporting both increased and decreased unpleasantness and emotional responsiveness in people with dementia compared to healthy controls. Clinically, depression is a significant burden for older people with dementia and chronic pain. The relationship between pain and other neuropsychiatric symptoms is controversial, and there is insufficient evidence on which to base conclusions. Some of the most important dementia-related changes may arise in the cognitive domain, including impairments of semantic and episodic memory for pain, executive function, and pain anticipation. Changes in brain activation and interconnectivity support many of these conclusions. Despite methodological limitations, we conclude there are compelling preliminary data to support a biopsychosocial framework of pain and dementia. Future research directions, especially the need for improved assessment tools, are highlighted.


Extracting product features and opinion words using pattern knowledge in customer reviews.
Due to the development of e-commerce and web technology, most of online Merchant sites are able to write comments about purchasing products for customer. Customer reviews expressed opinion about products or services which are collectively referred to as customer feedback data. Opinion extraction about products from customer reviews is becoming an interesting area of research and it is motivated to develop an automatic opinion mining application for users. Therefore, efficient method and techniques are needed to extract opinions from reviews. In this paper, we proposed a novel idea to find opinion words or phrases for each feature from customer reviews in an efficient way. Our focus in this paper is to get the patterns of opinion words/phrases about the feature of product from the review text through adjective, adverb, verb, and noun. The extracted features and opinions are useful for generating a meaningful summary that can provide significant informative resource to help the user as well as merchants to track the most suitable choice of product.


Follicle-restricted compartmentalization of transforming growth factor beta superfamily ligands in the feline ovary.
Ovarian follicular development, follicle selection, and the process of ovulation remain poorly understood in most species. Throughout reproductive life, follicle fate is balanced between growth and apoptosis. These opposing forces are controlled by numerous endocrine, paracrine, and autocrine factors, including the ligands represented by the transforming growth factor beta (TGFbeta) superfamily. TGFbeta, activin, inhibin, bone morphometric protein (BMP), and growth differentiation factor 9 (GDF-9) are present in the ovary of many animals; however, no comprehensive analysis of the localization of each ligand or its receptors and intracellular signaling molecules during folliculogenesis has been done. The domestic cat is an ideal model for studying ovarian follicle dynamics due to an abundance of all follicle populations, including primordial stage, and the amount of readily available tissue following routine animal spaying. Additionally, knowledge of the factors involved in feline follicular development could make an important impact on in vitro maturation/in vitro fertilization (IVM/IVF) success for endangered feline species. Thus, the presence and position of TGFbeta superfamily members within the feline ovary have been evaluated in all stages of follicular development by immunolocalization. The cat inhibin alpha subunit protein is present in all follicle stages but increases in intensity within the mural granulosa cells in large antral follicles. The inhibin betaA and betaB subunit proteins, in addition to the activin type I (ActRIB) and activin type II receptor (ActRIIB), are produced in primordial and primary follicle granulosa cells. Additionally, inhibin betaA subunit is detected in the theca cells from secondary through large antral follicle size classes. GDF-9 is restricted to the oocyte of preantral and antral follicles, whereas the type II BMP receptor (BMP-RII) protein is predominantly localized to primordial- and primary-stage follicles. TGFbeta1, 2, and 3 ligand immunoreactivity is observed in both small and large follicles, whereas the TGFbeta type II receptor (TGFbeta RII) is detected in the oocyte and granulosa cells of antral follicles. The intracellular signaling proteins Smad2 and Smad4 are present in the granulosa cell cytoplasm of all follicle size classes. Smad3 is detected in the granulosa cell nucleus, the oocyte, and the theca cell nucleus of all follicle size classes. These data suggest that the complete activin signal transduction pathway is present in small follicles and that large follicles primarily produce the inhibins. Our data also suggest that TGFbeta ligands and receptors are colocalized to large antral follicles. Taken together, the ligands, receptors, and signaling proteins for the TGFbeta superfamily are present at distinct points throughout feline folliculogenesis, suggesting discrete roles for each of these ligands during follicle maturation.


Proteomic Analysis of Sauvignon Blanc Grape Skin, Pulp and Seed and Relative Quantification of Pathogenesis-Related Proteins.
Thaumatin-like proteins (TLPs) and chitinases are the main constituents of so-called protein hazes which can form in finished white wine and which is a great concern of winemakers. These soluble pathogenesis-related (PR) proteins are extracted from grape berries. However, their distribution in different grape tissues is not well documented. In this study, proteins were first separately extracted from the skin, pulp and seed of Sauvignon Blanc grapes, followed by trypsin digestion and analysis by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Proteins identified included 75 proteins from Sauvignon Blanc grape skin, 63 from grape pulp and 35 from grape seed, mostly functionally classified as associated with metabolism and energy. Some were present exclusively in specific grape tissues; for example, proteins involved in photosynthesis were only detected in grape skin and proteins found in alcoholic fermentation were only detected in grape pulp. Moreover, proteins identified in grape seed were less diverse than those identified in grape skin and pulp. TLPs and chitinases were identified in both Sauvignon Blanc grape skin and pulp, but not in the seed. To relatively quantify the PR proteins, the protein extracts of grape tissues were seperated by HPLC first and then analysed by SDS-PAGE. The results showed that the protein fractions eluted at 9.3 min and 19.2 min under the chromatographic conditions of this study confirmed that these corresponded to TLPs and chitinases seperately. Thus, the relative quantification of TLPs and chitinases in protein extracts was carried out by comparing the area of corresponding peaks against the area of a thamautin standard. The results presented in this study clearly demonstrated the distribution of haze-forming PR proteins in grape berries, and the relative quantification of TLPs and chitinases could be applied in fast tracking of changes in PR proteins during grape growth and determination of PR proteins in berries at harvest.


[Conduction velocity and excitability of A and C fibers of the mesenteric nerve in the cat].
Conduction velocity and electrical excitability of mesenteric nerve fibres were studied in experiments on cats. These nerves were shown to consist of: 1) fibres with an excitation threshold of 0.06-0.10 V (stimuli duration being 0.1 ms) and with maximum conduction velocity of 48-85 m/s; 2) fibres with a threshold of 0.3-0.7 V and with conduction velocity from 8-10 to 33-39 7/s; 3) fibres with a threshold above 1 V and with conduction velocity from 1.8 to 7 m/s; 4) fibres with a threshold 6-8 V and conduction velocity below 1.8 m/s. The fibres of the first three groups are the afferent A-fibres. According to conduction velocity and wave amplitude in the compound action potential the fibres of the last group are divided into three subgroups. The first and third subgroups are shown to consist of afferent C-fibres; the second subgroup - of both afferent and postganglionic sympathetic C-fibres.


Prognostic benefit of extended radical lymphadenectomy for patients with gastric cancer.
The therapeutic value of extensive gastric lymphadenectomy in gastric cancer is controversial. We retrospectively investigated the effect of extended lymphadenectomy on survival in 882 patients with gastric cancer. From 882 patients who underwent gastrectomy, D0 or D1 lymphadenectomy was performed on 137 patients, D2 lymphadenectomy on 524 and D3 lymphadenectomy on 221. Curative gastrectomy was performed on 771 patients and the 5-year survival rate of patients had undergone D0 or D1 lymphadenectomy (D1 group) and that of patients who had undergone D2 lymphadenectomy (D2 group) was compared with that of patients had undergone D3 lymphadenectomy (D3 group). In each stage, the 5-year survival rates of patients who had undergone curative operations (n = 771) were compared among the D1, D2 and D3 groups. The 5-year survival rates were as follows: Stage I: n = 510, 89.3% (D1 group: n = 91, 85.8%; D2 group: n = 384, 90.3%; D3 group: n = 35, 88.1%; p = 0.539), Stage II: n = 83, 77.8% (D2 group: n = 35, 82.7%; D3 group: n = 48, 74.3%; p = 0.601), Stage III: n = 133, 50.2% (D2 group: n = 46, 39%; D3 group: n = 87, 56.1%; p = 0.027), Stage IV: n = 45, 8.9% (D2 group: n = 10, 0%; D3 group: n = 35, 11.4%; p = 0.588). Postoperative complications were detected in 10.4% of the cases and the in-hospital mortality rate was 2.4%. The postoperative morbidity rate of the D3 group (15.8%) was significantly higher than that of the other groups (D1 group: 7.3% and D2 group: 9%; p = 0.008). However, the in-hospital mortality rate of the D3 group (1.4%) was not different from that of the other groups (D1 group: 3.7% and D2 group: 2.5%; p = 0.374). These results indicate that D3 lymphadenectomy might be performed as safely as D1 or D2 lymphadenectomy on patients with gastric cancer. In addition, D3 lymphadenectomy might provide a survival benefit for patients with Stage III or Stage IV gastric cancer.


Diabetes Prevention in Australia: 10 Years Results and Experience.
Clinical trials have demonstrated the efficacy of lifestyle modification for the prevention of type 2 diabetes mellitus but it was achieved at higher cost than can be sustained in routine health services. The first clinical trial to report was the Finnish Diabetes Prevention Study. This paper describes how Australia worked with Finnish colleagues to adapt the findings of that study to achieve a statewide diabetes prevention program. Small evaluative, effectiveness trials have been conducted in a number of countries to see if the results of the clinical trials can be replicated in routine health services. The Australian evaluative trial, Greater Green Triangle Diabetes Prevention Program is described in detail to demonstrate the ingredients for success in moving a program from one country to another. Few countries have managed to scale up from evaluative trials to statewide or national programs. The Australian experience is described in detail including lessons learned about what reduced the effectiveness, particularly the need for policy makers in government, people from the implementing organisation and researchers to work together from the start of the evaluative trial and throughout the first 5 years of a national program.


The modulator of the general stress response, MgsR, of Bacillus subtilis is subject to multiple and complex control mechanisms.
The alternative sigma factor σ(B) is the master regulator of the general stress regulon that comprises approximately 200 genes whose products confer a comprehensive stress resistance to Bacillus subtilis. The characterization of MgsR (modulator of the general stress response) revealed that the activation and induction of σ(B) are a prerequisite but not sufficient for a full expression of all general stress genes. MgsR is a paralogue of the global regulator of the diamide stress response, Spx, and controls a subregulon of the general stress response. Here we demonstrate that MgsR activity is controlled at multiple levels. These mechanisms include a positive autoregulatory loop on mgsR transcription, a post-translational redox-sensitive activation step by an intramolecular disulfide bond formation in response to ethanol stress in vivo, as well as rapid proteolytic degradation of MgsR by the ClpXP and ClpCP proteases. Our results indicate an elaborate regulatory network integrating secondary oxidative stress signals into a σ(B) -mediated regulatory cascade that is aimed at rapid and finely tuned target gene expression to coordinately fulfil the physiological needs of the cell in the face of multiple environmental changes.


Architecture of peroxisomal alcohol oxidase crystals from the methylotrophic yeast Hansenula polymorpha as deduced by electron microscopy.
The architecture of alcohol oxidase crystalloids occurring in vivo in the peroxisomes of methylotrophic yeasts was deduced from electron micrographs of similar crystals of the Hansenula polymorpha enzyme grown in vitro. Three characteristic views of the crystal are observed, as well as single layers in the very early stages of crystal formation. The crystal is concluded to be cubical, with every octameric molecule making the same contacts with four neighbors in one plane, at right angles to its fourfold axis. The unit cell contains six octamers, in three mutually orthogonal orientations, and two large holes, which can accommodate other peroxisomal proteins involved in methanol metabolism. The crystal contains channels, connecting the holes, which allow the diffusion of relatively large molecules through the crystal. Crystal formation depends on just one contact per subunit, which may explain the fragility of the crystals.


Comparative activities of beta-lactam antibiotics and quinolones for invasive Streptococcus pneumoniae isolates.
Streptococcus pneumoniae is a leading pathogen causing pneumonia, meningitis, otitis media, bacteremia and sinusitis resulting in significant morbidity and mortality. We examined in vitro activities of five quinolones in comparison with other antibiotics against 85 invasive pneumococcal isolates. Minimal inhibitory concentrations (MICs) of penicillin G, cefuroxime, azithromycin, clarithromycin, trimethoprim-sulfamethoxazole (SXT), ciprofloxacin, ofloxacin, levofloxacin, trovafloxacin and gemifloxacin were determined using a broth microdilution method. The overall rates of resistance to penicillin (46%), cefuroxime (20%), azithromycin (20%), clarithromycin (18%) and SXT (46%) were considerable. Among all of the isolates, 9 isolates (11%) were highly resistant (MIC >/=2 mg/l) and 30 isolates (35%) had intermediate resistance (MIC 0.12- 1.0 mg/l). Of the quinolones gemifloxacin and trovafloxacin had the highest activity. The penicillin resistance status of the isolates did not have any effect on the resistance pattern of new quinolones. The new quinolones show great potential in the treatment of invasive infections caused by both penicillin-susceptible and penicillin-resistant pneumococci.


Comparative mechanistic and substrate specificity study of inositol polyphosphate 5-phosphatase Schizosaccharomyces pombe Synaptojanin and SHIP2.
Inositol-5-phosphatases are important enzymes involved in the regulation of diverse cellular processes from synaptic vesicle recycling to insulin signaling. We describe a comparative study of two representative inositol-5-phosphatases, Schizosaccharomyces pombe synaptojanin (SPsynaptojanin) and human SH2 domain-containing inositol-5-phosphatase SHIP2. We show that in addition to Mg2+, transition metals such as Mn2+, Co2+, and Ni2+ are also effective activators of SPsynaptojanin. In contrast, Ca2+ and Cu2+ are inhibitory. We provide evidence that Mg2+ binds the same site occupied by Ca2+ observed in the crystal structure of SPsynaptojanin complexed with inositol 1,4-bisphosphate (Ins(1,4)P2). Ionizations important for substrate binding and catalysis are defined for the SPsynaptojanin-catalyzed Ins(1,4,5)P3 reaction. Kinetic analysis with four phosphatidylinositol lipids bearing a 5-phosphate and 54 water-soluble inositol phosphates reveals that SP-synaptojanin and SHIP2 possess much broader substrate specificity than previously appreciated. The rank order for SPsynaptojanin is Ins(2,4,5)P3 > phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) approximately Ins(4,5)P2 approximately Ins(1,4,5)P3 approximately Ins(4,5,6)P3 > PtdIns(3,5)P2 approximately PtdIns(3,4,5)P3 approximately Ins(1,2,4,5)P4 approximately Ins(1,3,4,5)P4 approximately Ins-(2,4,5,6)P4 approximately Ins(1,2,4,5,6)P5. The rank order for SHIP2 is Ins(1,2,3,4,5)P5 > Ins(1,3,4,5)P4 > PtdIns(3,4,5)P4 approximately PtdIns(3,5)P2 approximately Ins(1,4,5,6)P4 approximately Ins(2,4,5,6)P4. Because inositol phosphate isomers elicit different biological activities, the extended substrate specificity for SPsynaptojanin and SHIP2 suggest that these enzymes likely have multiple roles in cell signaling and may regulate distinct pathways. The unique substrate specificity profiles and the importance of 2-position phosphate in binding also have important implications for the design of potent and selective SPsynaptojanin and SHIP2 inhibitors for pharmacological investigation.


Ultrasonic lens based on a subwavelength slit surrounded by grooves.
The lensing capabilities of a single subwavelength slit surrounded by a finite array of grooves milled into a brass plate is presented. The modulation of the beam intensity of this ultrasonic lens can be adjusted by varying the groove depth. Numerical simulations as well as experimental validations at 290 kHz are shown. The experimental results are in good agreement with the numerical simulations. This system is believed to have potential applications for medical ultrasound fields such as tomography and therapy.


A Sensitive and Rapid LC-MS-MS Method for Simultaneous Determination of Propafenone and Its Active Metabolite 5-Hydroxypropafenone in Human Plasma and Its Application in a Pharmacokinetic Study.
A simple and rapid high-performance liquid chromatography tandem mass spectrometry method for the determination of propafenone (PPF) and its active metabolite 5-hydroxypropafenone (5-OHP) in human plasma was developed and validated. This new method was linear and allowed simultaneous quantification of PPF and 5-OHP at a lower level of 0.5 ng/mL. The aliquot of 200 μL plasma sample was simply treated with 4-fold methanol to deproteinize the plasma. The chromatographic separation was achieved on a Hedera ODS-2C18 analytical column with the mobile phase of methanol and 5 mM ammonium acetate solution containing 0.2% formic acid (pH 3.2) (68:32, v/v) at a flow rate of 0.4 mL/min. Quantitation of the analytes was achieved by multiple reaction monitoring under positive ionization mode. The method was successfully applied to a pharmacokinetic study of PPF and 5-OHP in healthy Chinese volunteers. After oral administration of a single dose of 425 mg PPF hydrochloride sustained-release capsule, the maximum peak plasma concentration (Cmax) of PPF was 210.9 ± 141.9 ng/mL with a Tmax of 6 ± 1 h, the Cmax of 5-OHP was 129.6 ± 65.4 ng/mL with a Tmax of 7 ± 2 h. The area under plasma concentration-time curve (AUC0-36) of PPF was 1610 ± 1309 ng·h/mL with a t1/2 of 4.6 ± 1.1 h, the AUC0-36 of 5-OHP was 1446 ± 754 ng·h/mL with a t1/2 of 7.6 ± 1.6 h.


Molecular hydrogen as a preventive and therapeutic medical gas: initiation, development and potential of hydrogen medicine.
Molecular hydrogen (H2) has been accepted to be an inert and nonfunctional molecule in our body. We have turned this concept by demonstrating that H2 reacts with strong oxidants such as hydroxyl radical in cells, and proposed its potential for preventive and therapeutic applications. H2 has a number of advantages exhibiting extensive effects: H2 rapidly diffuses into tissues and cells, and it is mild enough neither to disturb metabolic redox reactions nor to affect signaling reactive oxygen species; therefore, there should be no or little adverse effects of H2. There are several methods to ingest or consume H2; inhaling H2 gas, drinking H2-dissolved water (H2-water), injecting H2-dissolved saline (H2-saline), taking an H2 bath, or dropping H2-saline into the eyes. The numerous publications on its biological and medical benefits revealed that H2 reduces oxidative stress not only by direct reactions with strong oxidants, but also indirectly by regulating various gene expressions. Moreover, by regulating the gene expressions, H2 functions as an anti-inflammatory and anti-apoptotic, and stimulates energy metabolism. In addition to growing evidence obtained by model animal experiments, extensive clinical examinations were performed or are under investigation. Since most drugs specifically act to their targets, H2 seems to differ from conventional pharmaceutical drugs. Owing to its great efficacy and lack of adverse effects, H2 has promising potential for clinical use against many diseases.


Betaine ameliorates impaired steroidogenesis and apoptosis in mice granulosa cells induced by high glucose concentration.
Betaine is a bioactive peptide whose beneficial effects on diabetes complications have been considered, previously. The present study aimed to investigate the possible protective effects of betaine against hyperglycemia-induced steroidogenesis impairment and apoptosis in mice granulosa cells. Ovarian granulosa cells were isolated from C57/BL6 mice and cultured in steroidogenesis medium (SM) containing 30 ng/ml FSH and 0.5 µM testosterone. The cells were cultured in SM containing low (5 mM) or high (30 mM) glucose concentrations for 24 h in the presence or absence of betaine (5 mM). At the end of the experiment, estradiol and progesterone were measured by ELISA in the culture medium. Expression of apoptosis and steroidogenesis associated genes and caspase-3 activity were determined by qRT-PCR and colorimetric assays, respectively. Exposure of mice granulosa cells to high glucose concentration inhibited the steroidogenesis by decreasing estradiol and progesterone secretion and downregulation of steroidogenesis-related genes including 3βHSD, Cyp11a1, Cyp19a1, and StAR. Betaine treatment could ameliorate the steroidogenesis impairment at molecular and biochemical levels. High glucose concentration also enhanced apoptosis in mice granulosa cells that were characterized by elevation of caspase-3 activity, upregulation of bax gene and downregulation of bcl2 gene. Betaine treatment could attenuate the apoptotic-related changes induced by high glucose concentration in granulosa cells. According to the results of the present study, betaine could ameliorate the adverse effects of hyperglycemia on the physiological function of ovarian granulosa cells. The results highlight the potential role of betaine for the intervention of ovarian dysfunction in diabetic patients. Abbreviations: AABA: Betaine-α-aminobutyric acid; AGEs: Advanced glycation end products; bax: bcl2 Associated X; bcl2: B-cell lymphoma 2; AMPK: AMP-activated protein kinase; BHMT: Betaine homocysteine methyltransferase; C/EBP: CCAAT-enhancer-binding proteins; Cyp11a1: Cholesterol side-chain cleavage cytochrome P450; Cyp19a1: Cytochrome P450 aromatase; DM: Diabetes mellitus; E2: Estradiol; ERS: Endoplasmic reticulum stress; GCs: Granulosa cells; GLUT: Glucose transporter; FSH: Follicle-stimulating hormone; 3βHSD: 3β-hydroxysteroid dehydrogenase; IL-1β: interleukin-1ß; LH: Luteinizing hormone; MDCK: Madin-Darby Canine Kidney cell; MT: Methionine synthase, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NLRP3: NLR Family Pyrin Domain Containing 3; NF-κB: Nuclear factor κB; P4: Progesterone; ROS: Reactive oxygen species; SGLT: Sodium dependent glucose transporter; SLC7A6: Solute Carrier Family 7 Member 6; StAR: Steroidogenic acute regulatory protein; STZ: Streptozotocin; Tumor necrosis factor α: TNF-α; TXNIP: Thioredoxin interacting protein.


In vitro systems for nephrotoxicity studies.
A variety of in vitro systems, including the perfused kidney, microperfused nephron(s), cultured renal slices, renal tubule suspensions/cultures, isolated nephron components (glomeruli and tubule segments) and renal cell suspensions/cultures can be utilized to evaluate chemicals for their potential nephrotoxicity and mechanisms of action. These systems have varying degrees of relevance to the intact kidney or portions thereof; which model to use depends on the specific question asked. If maintenance of physiological flow conditions is important then only the perfused kidney (or tubule) can be used. If the toxicity can be studied in the absence of normal fluid flow, then the other systems may be utilized. Kidney subfractions also have the advantage of multiple biological units that can be examined simultaneously and separately. Closely defined renal slice systems allow for enrichment and evaluation of multiple target cells within the slice. Suspensions or cultures of nephron subfractions are now frequently used to assess the potency and/or mechanism of renal toxins. Disadvantages include the utilization of collagenase, which scars cell and basement membranes and affects membrane transport proteins, which leads to inadequate maintenance of transport mechanisms. Isolation of tubule segments (P(1), P(2), P(3)), allows site-specific toxicity studies. Lastly, there has been a dramatic increase in renal cell culture to study toxic events in the kidney. Because of the multiple cell types, minimal availability of specific cell markers, and de-differentiation phenomena, this technique should be used with caution. These in vitro systems have allowed us to examine the role of transport and biotransformation, and to compare potency and mechanism of nephrotoxic chemicals. Although in vitro systems cannot totally duplicate or replace in vivo toxicity studies, they are a very useful and valuable complement.


Membrane Rigidity Determined by Atomic Force Microscopy Is a Parameter of the Permeability of Liposomal Membranes to the Hydrophilic Compound Calcein.
We determined the permeability coefficient of a model hydrophilic drug, calcein, encapsulated within saturated lipid-based nano-sized liposomes of various lipid profiles. We demonstrated that the addition of cholesterol to liposomes containing saturated lipids increased the permeability of the liposomal membrane to calcein via a decrease in the membrane bending modulus, as determined by means of atomic force microscopy. We found an inverse correlation between the membrane bending modulus of saturated lipid-based nano-sized liposomes and the permeability coefficient of encapsulated calcein, demonstrating that bending modulus, as determined by means of atomic force microscopy, is a quantitative parameter describing the permeability of liposomal membranes to calcein.


Central and storage carbon metabolism of the brown alga Ectocarpus siliculosus: insights into the origin and evolution of storage carbohydrates in Eukaryotes.
• Brown algae exhibit a unique carbon (C) storage metabolism. The photoassimilate D-fructose 6-phosphate is not used to produce sucrose but is converted into D-mannitol. These seaweeds also store C as β-1,3-glucan (laminarin), thus markedly departing from most living organisms, which use α-1,4-glucans (glycogen or starch). • Using a combination of bioinformatic and phylogenetic approaches, we identified the candidate genes for the enzymes involved in C storage in the genome of the brown alga Ectocarpus siliculosus and traced their evolutionary origins. • Ectocarpus possesses a complete set of enzymes for synthesis of mannitol, laminarin and trehalose. By contrast, the pathways for sucrose, starch and glycogen are completely absent. • The synthesis of β-1,3-glucans appears to be a very ancient eukaryotic pathway. Brown algae inherited the trehalose pathway from the red algal progenitor of phaeoplasts, while the mannitol pathway was acquired by lateral gene transfer from Actinobacteria. The starch metabolism of the red algal endosymbiont was entirely lost in the ancestor of Stramenopiles. In light of these novel findings we question the validity of the 'Chromalveolate hypothesis'.


Linking descriptive and experimental analyses in the treatment of bizarre speech.
Descriptive and experimental methods were used to analyze the environmental determinants of an adult's bizarre speech. A descriptive analysis of behavior under natural conditions indicated that bizarre vocalizations occurred most often in the presence of task-related demands and in the absence of adult attention. Further, bizarre speech occurring during tasks was followed frequently by the cessation of task demands by staff or the subject's voluntary disengagement from task-related activities; bizarre speech observed during noninteractional periods (i.e., in the absence of adult attention) was frequently followed by staff attention. The escape and attention hypotheses were tested under analogue conditions. Results of the experimental analysis supported only the attention hypothesis; that is, bizarre speech appeared to function as an attention-producing behavior. The functional analysis data were used to select two different yet functionally equivalent treatments. The first treatment provided the subject with noncontingent scheduled attention. The second intervention taught the subject social language skills in the form of initiation and expansion statements. Both interventions were effective in suppressing maladapted speech. Advantages of linking descriptive and experimental analyses are discussed.


Preoperative spirometry and laparotomy: blowing away dollars.
Increasing evidence indicates that routine preoperative diagnostic spirometry (pulmonary function tests [PFTs]) before elective abdominal surgery does not predict individual risk of postoperative pulmonary complications and is overutilized. This economic evaluation estimates potential savings from reduced use of preoperative PFTs. Analyses of (1) real costs (resource consumption to perform tests) and (2) reimbursements (expenditures for charges) by third-party payers. University-affiliated public and Veterans Affairs hospitals. Adults undergoing elective abdominal operations. Average real cost of PFTs was $19.07 (95% confidence interval [CI], $18.53 to $19.61), based on a time and motion study. Average reimbursement expenditure by third-party payers for PFTs was $85 (range, $33 to $150; 95% CI, $68 to $103), based on Medicare payment of $52 and a survey of nine urban US hospitals with a spectrum of bed sizes and teaching status. Estimates from published literature included the following: (1) annual number of major abdominal operations, 3.5 million; and (2) proportion of PFTs not meeting current guidelines, 39% (95% CI, 0.31 to 0.47). Local data were used when estimates were not available in the literature: (1) proportion of laparotomies that are elective, 76% (95% CI, 0.73 to 0.79); and (2) frequency of PFTs before laparotomy, 69% (95% CI, 0.54 to 0.84). Estimated annual national real costs for preoperative PFTs are $25 million to $45 million. If use of PFTs were reduced by our estimate for the proportion of PFTs not meeting current guidelines, potential annual national cost savings would be $7,925,411 to $21,406,707. National reimbursement expenditures by third-party payers range from more than $90 million to more than $235 million. If use were reduced, potential annual savings in reimbursements would be $29,084,076 to $111,345,440. Potential savings to Medicare approach $8 million to $20 million annually. Reduced use of PFTs before elective abdominal surgery could generate substantial savings. Current evidence indicates reduced use would not compromise patients' outcomes.


Environmental complexity: A buffer against stress in the domestic chick.
Birds kept in commercial production systems can be exposed to multiple stressors from early life and this alters the development of different morphological, immunological and behavioural indicators. We explore the hypothesis that provision of a complex environment during early life, better prepares birds to cope with stressful events as well as buffers them against future unpredictable stressful episodes. In this study, 96 one day old pullets were randomly distributed in eight pens (12 birds/pen). Half of the chicks (N = 48) were assigned to a Complex Environment (CENV: with perches, a dark brooder etc.) the others to a Simple Environment (SENV: without enrichment features). Half of the birds from each of these treatments were assigned to a No Stress (NSTR, 33°C) or to an acute Cold Stress (CSTR, 18-20°C) treatment during six hours on their second day of life. At four weeks of age, chicks with these four different backgrounds were exposed to an Intermittent Stressful Challenges Protocol (ISCP). In an immunological test indicative of pro-inflammatory status Phytohemagglutinin-P (PHA-P), the response of CSTR birds was ameliorated by rearing chicks in a CENV as they had a similar response to NSTR chicks and a significantly better pro-inflammatory response than those CSTR birds reared in a SENV (five days after the CSTR treatment was applied). A similar better response when coping with new challenges (the ISCP) was observed in birds reared in a CENV compared to those from a SENV. Birds reared in the CENV had a lower heterophil/lymphocyte ratio after the ISCP than birds reared in SENV, independently of whether or not they had been exposed to CSTR early in life. No effects of stress on general behaviour were detected, however, the provision of a CENV increased resting behaviour, which may have favoured stress recover. Additionally, we found that exposure to cold stress at an early age might have rendered birds more vulnerable to future stressful events. CSTR birds had lower humoral immune responses (sheep red blood cells induced antibodies) after the ISCP and started using elevated structures in the CENV later compared to their NSTR conspecifics. Our study reflects the importance of the early provision of a CENV in commercial conditions to reduce negative stress-related effects. Within the context of the theory of adaptive plasticity, our results suggest that the early experience of the birds had long lasting effects on the modulation of their phenotypes.


Role of Ca(2+)/calmodulin-dependent protein kinase kinase in adrenal aldosterone production.
There is considerable evidence supporting the role of calcium signaling in adrenal regulation of both aldosterone synthase (CYP11B2) and aldosterone production. However, there have been no studies that investigated the role played by the Ca(2+)/calmodulin-dependent protein kinase kinase (CaMKK) in adrenal cells. In this study we investigated the role of CaMKK in adrenal cell aldosterone production. To determine the role of CaMKK, we used a selective CaMKK inhibitor (STO-609) in the HAC15 human adrenal cell line. Cells were treated with angiotensin II (Ang II) or K+ and evaluated for the expression of steroidogenic acute regulatory protein and CYP11B2 (mRNA/protein) as well as aldosterone production. We also transduced HAC15 cells with lentiviral short hairpin RNAs of CaMKK1 and CaMKK2 to determine which CaMKK plays a more important role in adrenal cell regulation of the calcium signaling cascade. The CaMKK inhibitor, STO-609, decreased aldosterone production in cells treated with Ang II or K+ in a dose-dependent manner. STO-609 (20 μM) also inhibited steroidogenic acute regulatory protein and CYP11B2 mRNA/protein induction. CaMKK2 knockdown cells showed significant reduction of CYP11B2 mRNA induction and aldosterone production in cells treated with Ang II, although there was no obvious effect in CaMKK1 knockdown cells. In immunohistochemical analysis, CaMKK2 protein was highly expressed in human adrenal zona glomerulosa with lower expression in the zona fasciculata. In conclusion, the present study suggests that CaMKK2 plays a pivotal role in the calcium signaling cascade regulating adrenal aldosterone production.


Pyelonephritis, cortical abscess, and perinephric abscess.
The causative agent in nonobstructive pyelonephritis has been shown most often to be P-fimbriated Escherichia coli, mainly because receptors for these fimbriae are found in the bladder, ureter, and the kidney tubules. Age and sex are factors leading to differences in the presentation of the disease, and early diagnosis followed by intensive therapy lessens the chance of renal damage. Renal abscess, perinephric abscess, and pyonephrosis all follow pyelonephritis in the host who is compromised by the presence of stone, obstruction, diabetes, or immunosuppression.


Toxoplasma gondii: immune response and protective efficacy induced by ROP16/GRA7 multicomponent DNA vaccine with a genetic adjuvant B7-2.
Toxoplasma gondii infection occurs commonly in humans and other warm-blooded animals. Its serious impact on public health and livestock sectors makes the development of an effective vaccine particularly important. In the current study, we constructed a multiantigenic DNA vaccine expressing ROP16 and GRA7 of T. gondii and evaluated the protective efficacy of these two fragments with or without a plasmid encoding murine costimulatory molecule B7-2. These recombinant eukaryotic expression plasmids were termed pROP16, pGRA7, pROP16-GRA7 and pB7-2, respectively. After intramuscular immunization in Kunming mice, we assessed the immune response using cytokine and antibody determinations, T lymphocyte subsets analysis, and the survival times of mice post acute T. gondii challenge. The results showed that mice immunized with the multiantigenic DNA vaccine pROP16-GRA7 gained higher levels of IgG titers and IgG2a subclass titers, production of IFN-γ, percentage of CD8+ T cells and median survival times against the acute infection of T. gondii compared with those of mice administered with pROP16 or pGRA7 and those in control groups. Moreover, the adjuvant pB7-2 formulated with DNA vaccine boosted these humoral and cellular (Th1, CD8+ T cell) immune responses. Therefore, it might be a promising genetic adjuvant to DNA vaccine against T. gondii for further investigation.


Temporal adaptation of neutrophil oxidative responsiveness to n-formyl-methionyl-leucyl-phenylalanine. Acceleration by granulocyte-macrophage colony stimulating factor.
This investigation was undertaken to clarify the mechanism by which purified recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) potentiates neutrophil oxidative responses triggered by the chemotactic peptide, FMLP. Previous studies have shown that GM-CSF priming of neutrophil responses to FMLP is induced relatively slowly, requiring 90 to 120 min of incubation in vitro, is not associated with increased levels of cytoplasmic free Ca2+, but is associated with up-regulation of cell-surface FMLP receptors. We have confirmed these findings and further characterized the process of GM-CSF priming. We found that the effect of GM-CSF on neutrophil oxidative responsiveness was induced in a temperature-dependent manner and was not reversed when the cells were washed extensively to remove the growth factor before stimulation with FMLP. Extracellular Ca2+ was not required for functional enhancement by GM-CSF and GM-CSF alone effected no detectable alteration in the 32P-labeled phospholipid content of neutrophils during incubation in vitro. Our data indicate that GM-CSF exerts its influence on neutrophils by accelerating a process that occurs spontaneously and results in up-regulation of both cell-surface FMLP receptors and oxidative responsiveness to FMLP. Thus, the results demonstrate that, with respect to oxidative activation, circulating endstage polymorphonuclear leukocytes are nonresponsive or hyporesponsive to FMLP; functional responsiveness increases dramatically as surface FMLP receptors are gradually deployed after the cells leave the circulation. Thus, as neutrophils mature, their responsiveness to FMLP changes in a manner which may be crucial for efficient host defense. At 37 degrees C, this process is markedly potentiated by GM-CSF. We conclude that endogenous GM-CSF, released systemically or at sites of infection and inflammation, potentially plays an important role in host defense by accelerating functional maturation of responding polymorphonuclear leukocytes.


Zebrafish approaches enhance the translational research tackle box.
During the past few decades, zebrafish (Danio rerio) have been a workhorse for developmental biology and genetics. Concurrently, zebrafish have proved highly accessible and effective for translational research by providing a vertebrate animal model useful for gene discovery, disease modeling, chemical genetic screening, and other medically relevant studies. Key resources such as an annotated and complete genome sequence, and diverse tools for genetic manipulation continue to spur broad use of zebrafish. Thus, the purpose of this introductory review is to provide a window into the unique characteristics and diverse uses of zebrafish and to highlight in particular the increasing relevance of zebrafish as a translational animal model. This is accomplished by reviewing broad considerations of anatomic and physiological conservation, approaches for disease modeling and creation, general laboratory methods, genetic tools, genome conservation, and diverse opportunities for functional validation. Additional commentary throughout the review also guides the reader to the 4 new reviews found elsewhere in this special issue that showcase the many unique ways the zebrafish is improving understanding of renal regeneration, mitochondrial disease, dyslipidemia, and aging, for example. With many other possible approaches and a rapidly increasing number of medically relevant reports, zebrafish approaches enhance significantly the tools available for translational research and are actively improving the understanding of human disease.


Effects of glucose, pyruvate, lactate, and amino acids on muscle protein synthesis.
Protein synthesis was measured in rat diaphragms incubated with serum amino acids + 0.35 mM L-[2,6-3H]tyrosine and different energy-yielding substrates. Muscles incubated with 5.5 mM glucose (with or without actinomycin D) synthesized more protein than those incubated with 11 mM pyruvate or 11 mM lactate. Tissue ATP decreased during incubation with lactate, but pyruvate maintained ATP, ADP, and creatine phosphate as well as glucose. Glucose 6-phosphate decreased in muscles incubated in glucose-free media. 14CO2 production from substrates was [1-14C]pyruvate greater than [1-14C]lactate greater than [3,4-14C]glucose. Intracellular lactate/pyruvate was measured to assess cytoplasmic free NADH/NAD+; the effect of different media on these ratios was lactate greater than glucose = lactate + pyruvate greater than pyruvate + glucose greater than pyruvate. Lactate + pyruvate (8.8 + 2.2 mM) supported protein synthesis better than pyruvate and as well as glucose. Adding glucose to pyruvate accelerated protein synthesis and increased NADH/NAD+. Iodoacetate (0.1 mM) inhibited glycolytic NAD reduction and abolished the stimulatory effect of glucose on protein synthesis in the presence of pyruvate. Supplementation of pyruvate media with 1 mM leucine or isoleucine stimulated protein synthesis, but beta-hydroxybutyrate, malate, alpha-ketoisocaproate, and all other amino acids were ineffective. The cytoplasmic redox potential may act as a translational modulator of protein synthesis in skeletal muscle.


Unveiling synapse pathology in spinal bulbar muscular atrophy by genome-wide transcriptome analysis of purified motor neurons derived from disease specific iPSCs.
Spinal bulbar muscular atrophy (SBMA) is an adult-onset, slowly progressive motor neuron disease caused by abnormal CAG repeat expansion in the androgen receptor (AR) gene. Although ligand (testosterone)-dependent mutant AR aggregation has been shown to play important roles in motor neuronal degeneration by the analyses of transgenic mice models and in vitro cell culture models, the underlying disease mechanisms remain to be fully elucidated because of the discrepancy between model mice and SBMA patients. Thus, novel human disease models that recapitulate SBMA patients' pathology more accurately are required for more precise pathophysiological analysis and the development of novel therapeutics. Here, we established disease specific iPSCs from four SBMA patients, and differentiated them into spinal motor neurons. To investigate motor neuron specific pathology, we purified iPSC-derived motor neurons using flow cytometry and cell sorting based on the motor neuron specific reporter, HB9e438::Venus, and proceeded to the genome-wide transcriptome analysis by RNA sequences. The results revealed the involvement of the pathology associated with synapses, epigenetics, and endoplasmic reticulum (ER) in SBMA. Notably, we demonstrated the involvement of the neuromuscular synapse via significant upregulation of Synaptotagmin, R-Spondin2 (RSPO2), and WNT ligands in motor neurons derived from SBMA patients, which are known to be associated with neuromuscular junction (NMJ) formation and acetylcholine receptor (AChR) clustering. These aberrant gene expression in neuromuscular synapses might represent a novel therapeutic target for SBMA.


Atmospheric pressure plasma induced grafting of poly(ethylene glycol) onto silicone elastomers for controlling biological response.
This study investigates the role that surface functionalisation of silicone elastomer (SE) by atmospheric pressure plasma induced graft immobilisation of poly(ethylene glycol) methyl ether methacrylate (PEGMA) plays in the attendant biological response. SE is used in modern ophthalmic medical devices and samples of the material were initially plasma treated using a dielectric barrier discharge reactor (DBD) to introduce reactive oxygen functionalities, prior to in situ grafting of two molecular weights of PEGMA (MW 1000 Da: PEGMA(1000), MW 2000 Da: PEGMA(2000)). The variously processed surfaces were characterised by water contact angle analysis, X-ray photoelectron spectroscopy, time-of-flight secondary ion mass spectrometry and atomic force microscopy. Lens epithelial cells were then cultured on the PEGMA grafted SE surfaces. It was found that cells on the pristine surface were not well spread and had shrunken morphology. On the DBD pre-treated surfaces, the cells were well spread. On the PEGMA(1000) surface, the cells displayed evidence of shrinkage and were on the verge of detaching. Remarkably, on the PEGMA(2000) surface, no cell adhesion was detection. Bacterial adhesion to the surfaces was studied using Staphylococcus aureus NTC8325. There was no difference in the number of bacteria adhering to any of the surfaces studied.


Mapping of neuroinflammation-induced hypoxia in the spinal cord using optoacoustic imaging.
Recent studies suggest that metabolic changes and oxygen deficiency in the central nervous system play an important role in the pathophysiology of multiple sclerosis (MS). In our present study, we investigated the changes in oxygenation and analyzed the vascular perfusion of the spinal cord in a rodent model of MS. We performed multispectral optoacoustic tomography of the lumbar spinal cord before and after an oxygen enhancement challenge in mice with experimental autoimmune encephalomyelitis (EAE), a model for MS. In addition, mice were transcardially perfused with lectin to label the vasculature and their spinal columns were optically cleared, followed by light sheet fluorescence microscopy. To analyze the angioarchitecture of the intact spine, we used VesSAP, a novel deep learning-based framework. In EAE mice, the spinal cord had lower oxygen saturation and hemoglobin concentration compared to healthy mice, indicating compromised perfusion of the spinal cord. Oxygen administration reversed hypoxia in the spinal cord of EAE mice, although the ventral region remained hypoxic. Additionally, despite the increased vascular density, we report a reduction in length and complexity of the perfused vascular network in EAE. Taken together, these findings highlight a new aspect of neuroinflammatory pathology, revealing a significant degree of hypoxia in EAE in vivo that is accompanied by changes in spinal vascular perfusion. The study also introduces optoacoustic imaging as a tractable technique with the potential to further decipher the role of hypoxia in EAE and to monitor it in MS patients.


Coordination of transcription factor phosphorylation and histone methylation by the P-TEFb kinase during human immunodeficiency virus type 1 transcription.
The human immunodeficiency virus type 1 (HIV-1) Tat protein recruits positive transcription elongation factor b (P-TEFb) to the transactivation response (TAR) RNA structure to facilitate formation of processive transcription elongation complexes (TECs). Here we examine the role of the Tat/TAR-specified cyclin-dependent kinase 9 (CDK9) kinase activity in regulation of HIV-1 transcription elongation and histone methylation. In HIV-1 TECs, P-TEFb phosphorylates the RNA polymerase II (RNAP II) carboxyl-terminal domain (CTD) and the transcription elongation factors SPT5 and Tat-SF1 in a Tat/TAR-dependent manner. Using in vivo chromatin immunoprecipitation analysis, we demonstrate the following distinct properties of the HIV-1 transcription complexes. First, the RNAP II CTD is phosphorylated at Ser 2 and Ser 5 near the promoter and at downstream coding regions. Second, the stable association of SPT5 with the TECs is dependent upon P-TEFb kinase activity. Third, P-TEFb kinase activity is critical for the induction of methylation of histone H3 at lysine 4 and lysine 36 on HIV-1 genes. Flavopiridol, a potent P-TEFb kinase inhibitor, inhibits CTD phosphorylation, stable SPT5 binding, and histone methylation, suggesting that its potent antiviral activity is due to its ability to inhibit several critical and unique steps in HIV-1 transcription elongation.


Thermococcus eurythermalis sp. nov., a conditional piezophilic, hyperthermophilic archaeon with a wide temperature range for growth, isolated from an oil-immersed chimney in the Guaymas Basin.
A conditional piezophilic, hyperthermophilic archaeon showing growth over a wide range of temperature, pH and pressure was isolated from an oil-immersed hydrothermal chimney at a depth of 2006.9 m in the Guaymas Basin. Enrichment and isolation of strain A501(T) were performed at 80 °C at 0.1 MPa. Cells of isolate A501(T) were irregular motile cocci with a polar tuft of flagella and generally 0.6-2.6 µm in diameter. Growth was detected over the range 50-100 °C (optimal growth at 85 °C) at atmospheric pressure and was observed at 102 °C at a pressure of 10 MPa. At 85 °C, growth was observed at a pressure of 0.1-70 MPa (optimum pressure 0.1 MPa-30 MPa), while at 95 °C, the pressure allowing growth ranged from 0.1 MPa to 50 MPa (optimum pressure 10 MPa). Cells of strain A501(T) grew at pH 4-9 (optimum pH 7.0) and a NaCl concentration of 1.0-5.0 % (w/v) (optimum concentration 2.5 % NaCl). This isolate was an anaerobic chemo-organoheterotroph and was able to utilize yeast extract, peptone, tryptone and starch as the single carbon source for growth. Elemental sulfur and cysteine stimulated growth; however, these molecules were not necessary. The DNA G+C content of the complete genome was 53.47 mol%. The results of 16S rRNA gene sequence analysis indicated that strain A501(T) belongs to the genus Thermococcus. There was no significant similarity between strain A501(T) and the phylogenetically related species of the genus Thermococcus based on complete genome sequence alignments and calculation of the average nucleotide identity and the tetranucleotide signature frequency correlation coefficient. These results indicate that strain A501(T) represents a novel species, Thermococcus eurythermalis sp. nov. The type strain is A501(T) ( = CGMCC 7834(T) = JCM 30233(T)).


A Test in Context: D-Dimer.
D-dimer is a soluble fibrin degradation product that results from ordered breakdown of thrombi by the fibrinolytic system. Numerous studies have shown that D-dimer serves as a valuable marker of activation of coagulation and fibrinolysis. Consequently, D-dimer has been extensively investigated for the diagnosis of venous thromboembolism (VTE) and is used routinely for this indication. In addition, D-dimer has been evaluated for determining the optimal duration of anticoagulation in VTE patients, for diagnosing and monitoring disseminated intravascular coagulation, and as an aid in the identification of medical patients at high risk for VTE. Thus, quantification of D-dimer levels serves an important role in guiding therapy. This review: 1) describes how D-dimer is generated; 2) reviews the assays used for its detection; and 3) discusses the role of D-dimer determination in these various conditions.


Predictive Impact of Clinical Benefit in Chemotherapy-treated Advanced Pancreatic Cancer Patients in Northern Alberta.
Patients with advanced pancreatic cancer (APC) have a poor prognosis and experience a large burden of disease-related symptoms. Despite advancements in the treatment of APC, survival is dismal and controlling disease-related symptoms and maintaining quality of life is paramount. We hypothesize that an improvement in disease-related symptoms, and therefore, a clinical benefit, while on chemotherapy is a predictive marker in APC. Patients 18 and older with APC diagnosed between January 1, 2005 and December 31, 2010 and treated at the Cross Cancer Institute were identified using the provincial cancer registry. Disease symptoms were assessed at baseline and clinical benefit while on chemotherapy was defined using a composite endpoint of improvement in patient-reported pain, opioid consumption, Eastern Cooperative Oncology Group performance status, and/or weight. Best radiologic response, progression-free survival (PFS), and overall survival (OS) were recorded. Of 103 patients, the median age was 64, 58% were male and 66% had metastatic disease. At baseline, the majority of patients reported pain (80%), opioid use (61%), or weight loss (71%). In total, 35 (34%) patients received a clinical benefit with treatment but only 6 (17%) of these patients experienced a radiologic response. The median PFS and OS were improved in patients who experienced a clinical benefit (6.6 vs. 4.6 mo; P=0.03 and 11.7 vs. 6.1 mo; P<0.0001, respectively). In patients with APC treated with chemotherapy, experiencing a clinical benefit was associated with improved PFS and OS. However, it did not appear to correlate with radiologic response to chemotherapy. Prospective studies are warranted to further investigate the prognostic and predictive value of clinical benefit and improvement in quality of life as measured by standardized tools, in APC.


Assessment of craniocervical junction and atlantoaxial relation using metrizamide-enhanced CT in flexion and extension.
Metrizamide-enhanced computed tomographic (CT) myelography has made it easier to define the relation of the spinal cord to the vertebral canal. A flexion-extension metrizamide-enhanced CT technique has been developed to study the craniocervical junction that refines evaluation of the relation between the spinal cord and the foramen magnum, atlas, and axis. This technique was used to study 15 adults who had had a structurally normal examination of the upper cervical cord and foramen magnum. The average movement of the upper cervical cord was shown to be 1 mm. The advantages of the flexion-extension metrizamide-enhanced CT examination were evident in 10 other patients who had a variety of craniocervical junction pathologies.


AKT/protein kinase B associates with β-actin in the nucleus of melanoma cells.
The serine-threonine kinase AKT/PKB is a critical regulator of various essential cellular processes, and dysregulation of AKT has been implicated in many diseases, including cancer. Despite AKT action is known to function mainly in the cytoplasm, AKT has been reported to translocate to the nucleus. However, very little is known about the mechanism required for the nuclear import of AKT as well as its function in this cellular compartment. In the present study, we characterized the presence of endogenous nuclear AKT in human melanoma cells and addressed the possible role of AKT by exploring its potential association with key interaction nuclear partners. Confocal and Western blot analyses showed that both phosphorylated and non-phosphorylated forms of AKT are present in melanoma cells nuclei. Using mass spectrometry in combination with protein-crosslinking and co-immunoprecipitation, we identified a series of putative protein partners of nuclear AKT, including heterogeneous nuclear ribonucleoprotein (hnRNP), cytoskeleton proteins β-actin, γ-actin, β-actin-like 2 and vimentin. Confocal microscopy and biochemical analyses validated β-actin as a new nuclear AKT-interacting partner. Cofilin and active RNA Polymerase II, two proteins that have been described to interact and work in concert with nuclear actin in transcription regulation, were also found associated with nuclear AKT. Overall, the present study uncovered a yet unrecognized nuclear coupling of AKT and provides insights into the involvement of AKT in the interaction network of nuclear actin.


Impact of pulmonary hypertension on long-term outcome in patients with severe aortic stenosis.
Pulmonary hypertension (PH) is common in severe symptomatic left-sided valvular disease, particularly in aging populations. Inconsistent results have been reported concerning the association between PH and adverse outcomes after aortic valve replacement for aortic stenosis (AS). We therefore retrospectively investigated the prognostic significance of PH using peak tricuspid regurgitation velocity (TRV), as defined by the current European Society of Cardiology (ESC)/European Respiratory Society (ERS) guidelines, in a large cohort of patients with severe AS. One thousand and nineteen patients (541 men; mean age 74 ± 11 years) with severe AS (aortic valve area (AVA) <1 cm2 and/or indexed AVA <0.6 cm2/m2 of body surface area) and LV ejection fraction ≥50% were included. Patients were divided into three groups according to the level of their peak TRV at the time of enrolment: Group 1 (n = 695, 68%) when TRV was ≤2.8 m/s; Group 2 (n = 212, 21%) when TRV was between 2.9 m/s and 3.4 m/s and Group 3 (n = 112, 11%) when TRV was > 3.4 m/s. Median overall follow-up was 31 [6-182] months. On univariate analysis, overall mortality during follow-up was globally different between groups (P < 0.001). On multivariate analysis, Group 3 (TRV >3.4 m/s) exhibited significant excess mortality after adjustment for covariates of prognostic importance (P = 0.032) and after further adjustment for surgery (P = 0.012), using Group 1 as the reference group. Dividing the whole population into two groups with a 3.4 m/s TRV threshold, overall mortality during follow-up was higher in the PH group [hazard ratio (HR) 1.87; 95% confidence interval [1.37-2.56]; P < 0.001)]. On multivariate analysis, after covariate adjustment, including surgery, Group 3 exhibited major excess mortality (adjusted HR 1.46 [1.10-1.95], P = 0.009). This study demonstrates the negative impact of pulmonary pressure, as assessed by current ESC/ERS guidelines, on long-term outcome of patients with severe AS, irrespective of functional status, chronic obstructive pulmonary disease, AS severity and surgery. Baseline TRV should therefore be taken into account in the management of severe AS.


Molecular genetic diversity of Gongylonema neoplasticum (Fibiger & Ditlevsen, 1914) (Spirurida: Gongylonematidae) from rodents in Southeast Asia.
More than a dozen Gongylonema spp. (Spirurida: Spiruroidea: Gongylonematidae) have been described from a variety of rodent hosts worldwide. Gongylonema neoplasticum (Fibiger & Ditlevsen, 1914), which dwells in the gastric mucosa of rats such as Rattus norvegicus (Berkenhout) and Rattus rattus (Linnaeus), is currently regarded as a cosmopolitan nematode in accordance with global dispersion of its definitive hosts beyond Asia. To facilitate the reliable specific differentiation of local rodent Gongylonema spp. from the cosmopolitan congener, the genetic characterisation of G. neoplasticum from Asian Rattus spp. in the original endemic area should be considered since the morphological identification of Gongylonema spp. is often difficult due to variations of critical phenotypical characters, e.g. spicule lengths and numbers of caudal papillae. In the present study, morphologically identified G. neoplasticum from 114 rats of seven species from Southeast Asia were selected from archived survey materials from almost 4,500 rodents: Thailand (58 rats), Cambodia (52 rats), Laos (three rats) and Philippines (one rat). In addition, several specimens from four rats in Indonesia were used in the study. Nucleotide sequences of the ribosomal RNA gene (rDNA) (5,649 bp) and the cytochrome c oxidase subunit 1 gene (cox1) (818 bp) were characterised. The rDNA showed little nucleotide variation, including the internal transcribed spacer (ITS) regions. The cox1 showed 24 haplotypes, with up to 15 (1.83%) nucleotide substitutions regardless of parasite origin. Considering that Rattus spp. have been shown to originate from the southern region of Asia and G. neoplasticum is their endogenous parasite, it is reasonable to propose that the present study covers a wide spectrum of the genetic diversity of G. neoplasticum, useful for both the molecular genetic speculation of the species and the molecular genetic differentiation of other local rodent Gongylonema spp. from the cosmopolitan congener.


Adult participation in children's word searches: on the use of prompting, hinting, and supplying a model.
Although word searching in children is very common, very little is known about how adults support children in the turns following the child's search behaviours, an important topic because of the social, educational, and clinical implications. This study characterizes, in detail, teachers' use of prompting, hinting, and supplying a model. From a classroom dataset of 53 instances, several distinctive patterns emerged. A prompted completion sequence is initiated by a 'word retrieval elicitor' ('fishing::') and is interpreted as a request to complete the phrase. Non-verbal prompting is accomplished through a combination of gaze and gesture and, also, as a series of prompts. Hinting supplies a verbal clue, typically via a wh-question, or by specifying the nature of the repairable. In contrast, the strategies that supply a linguistic model include both embedded and exposed corrections and offers of candidates. A sequential relationship was found between prompting, hinting, and supplying a model which has implications for how clinicians and teachers can foster self-repair.


Effect of drought and nutrient availability on invaded plant communities in a semi-arid ecosystem.
Ecosystem functions are heavily dependent on the functional composition of the plant community, i.e., the functional traits of plants forming the community. This, on the one hand, depends on plant occurrence, but on the other hand, depends on the intraspecific variability of functional traits of the species, which are influenced by climate and nutrient availability and affected by plant-plant interactions. To illustrate that, we studied the effects of drought and nitrogen addition (+ N), two important abiotic variables which are changing with ongoing global change, as well as their combined effect on the functional responses of grassland communities in semi-arid environments of Northern Africa comprising of natural and invasive species. We conducted an experiment where we planted three native species and one invasive plant species in artificial communities of five individuals per species per plot. We exposed these communities to four different treatments: a drought treatment, an N-addition treatment, the combination between drought and N-addition, as well as a control. To assess the performance of plants within treatments, we measured selected plant functional traits (plant height, specific leaf area [SLA], leaf dry matter content [LDMC], N content of the leaves [Nmass], specific root length [SRL], and root diameter) for all individuals occurring in our plots, and additionally assessed the above and belowground biomass for each plant individual. We found that the invasive species showed a higher performance (higher biomass accumulation, taller plants, higher SLA, Nmass, SRL, and root diameter as well as lower LDMC) than the native species under drought conditions. The invasive species was especially successful with the combined impact of drought + N, which is a likely scenario in ongoing global change for our research area. Thus, plant functional traits might be a key factor for the invasion success of plant species which will be even more pronounced under ongoing global change.


Retinoic acid is a key regulatory switch determining the difference between lung and thyroid fates in Xenopus laevis.
The lung and thyroid are derived from the anterior endoderm. Retinoic acid and Fgf signalling are known to be essential for development of the lung in mouse but little is known on how the lung and thyroid are specified in Xenopus. If either retinoic acid or Fgf signalling is inhibited, there is no differentiation of the lung as assayed by expression of sftpb. There is no change in expression of thyroid gland markers when retinoic acid signalling is blocked after gastrulation and when Fgf signalling is inhibited there is a short window of time where pax2 expression is inhibited but expression of other markers is unaffected. If exogenous retinoic acid is given to the embryo between embryonic stages 20 and 26, the presumptive thyroid expresses sftpb and sftpc, specific markers of lung differentiation and expression of key thyroid transcription factors is lost. When the presumptive thyroid is transplanted into the posterior embryo, it also expresses sftpb, although pax2 expression is not blocked. After gastrulation, retinoic acid is required for lung but not thyroid differentiation in Xenopus while Fgf signalling is needed for lung but only for early expression of pax2 in the thyroid. Exposure to retinoic acid can cause the presumptive thyroid to switch to a lung developmental program.


Comparative in vitro activity of tigecycline and other antimicrobial agents against Shigella species from Kuwait and the United Arab of Emirates.
Shigella species isolated from stool samples of symptomatic patients of all age groups at the Mubarak Al Kabir Hospital and Infectious Diseases Hospital, Kuwait and Tawam Hospital, UAE during a 2-year period were investigated for their susceptibility to tigecycline and several other antibiotics by determining the minimum inhibitory concentrations (MICs) using the E test method. A total of 100 and 42 strains were collected from UAE and Kuwait, respectively. The extent of drug resistance in the Shigella spp. isolates from these two countries was analyzed by criteria recommended by the Clinical and Laboratory Standards Institute (CLSI). Amikacin, cefotaxime, cefuroxime, ciprofloxacin, imipenem, meropenem, piperacillin-tazobactam and tigecycline had excellent activities against all isolates from UAE and Kuwait with MIC(90s) of 12, 0.094, 4, 0.012, 0.25, 0.032, 3 and 0.25 microg/ml and 4, 1, 4, 0.125, 0.38, 0.19, 3 and 0.25 microg/ml, respectively. Half of all isolates from both countries were resistant to ampicillin. None of the isolates in Kuwait was resistant to amoxicillin-clavulanic acid compared with 22% in UAE. Resistance to chloramphenicol was recorded in 50 and 36% of the isolates in Kuwait and UAE, respectively. The percentages of non-susceptibility to trimethoprim-sulfamethoxazole and tetracycline were very high in Kuwait and UAE (76% vs. 92% and 76% vs. 98%, respectively). Notably, one isolate, S. flexneri, from UAE had reduced susceptibility to ciprofloxacin (MIC, 0.25 microg/ml). Four (2.8%) of the isolates were ESBL producers by the E test ESBL method but could not be confirmed by PCR using primers for bla(CTX-M), bla(SHV) and bla(TEM). In conclusion, Shigella spp. isolated from symptomatic patients in Kuwait and the UAE demonstrated high rates of resistance to the first-line antibiotics but very susceptible to the carbapenems, cephalosporins, fluoroquinolones and tigecycline. Tigecycline holds promise as a potential drug of choice for the therapy of severe shigellosis.


Basolateral potassium channels in renal proximal tubule.
Potassium (K+) channels in the basolateral membrane of unperfused Necturus proximal tubules were studied in both cell-attached and excised patches, after removal of the tubule basement membrane by manual dissection without collagenase. Two different K+ channels were identified on the basis of their kinetics: a short open-time K+ channel, with a mean open time less than 1 ms, and a long open-time K+ channel with a mean open time greater than 20 ms. The short open-time channel occurred more frequently than the longer channel, especially in excised patches. For inside-out excised patches with Cl- replaced by gluconate, the current-voltage relation of the short open-time K+ channel was linear over +/- 60 mV, with a K+-Na+ selectivity of 12 +/- 2 (n = 12), as calculated from the reversal potential with oppositely directed Na+ and K+ gradients. With K-Ringer in the patch pipette and Na-Ringer in the bath, the conductance of the short open-time channel was 47 +/- 2 pS (n = 15) for cell-attached patches, 26 +/- 2 pS (n = 15) for patches excised (inside out) into Na-Ringer, and 36 +/- 6 pS (n = 3) for excised patches with K-Ringer on both sides. These different conductances can be partially explained by a dependence of single-channel conductance on the K+ concentration on the interior side of the membrane. In experiments with a constant K+ gradient across excised patches, large changes in Na+ at the interior side of the membrane produced no change in single-channel conductance, arguing against a direct block of the K+ channel by Na+. Finally, the activity of the short open-time channel was voltage gated, where the mean number of open channels decreased as a linear function of basolateral membrane depolarization for potentials between -60 and 0 mV. Depolarization from -60 to -40 mV decreased the mean number of open K+ channels by 28 +/- 8% (n = 6).


Reciprocal relationship between the major EEG rhythms and latencies of evoked potential intermediate components. A tentative explanation.
We recently demonstrated that there is a reciprocal relationship between the known brain wave rhythms (delta, 3.3 c/sec, 5 c/sec, theta, alpha, spindle, and beta) on the one hand, and the averaged evoked potential mid component latencies, on the other. In other words, the latter are closely related to the periods of the known brain wave frequencies. In this paper we offer a hypothetical explanation of this puzzling relationship, by assuming the following: 1) Whenever the fastest component of a sensory signal reaches the cortex it depresses EEG except for those waves of different frequencies which at that very instant exhibit a high amplitude peak of a preferential polarity. One half period later, different in real time for each brain wave frequency, high amplitude peaks will emerge from the record with opposite polarity to that which was preferential at the time of the signals' arrival. These segmental peaks are the same as those that express evoked potential components of mid latencies. For the somatosensory evoked potentials the time of occurrence of these peaks are defined by the following expression: (Formula; see text) where N is equal to successive numbers between 1 and 12, as well as to the fractions of 1; 20 msec is the moment of arrival of the fastest signal to the cortex. Some of the latencies correspond to the arrival of the delayed components of the somatosensory signals. Some correspond to the time of arrival of the auditory and visual signals, in an apparent expectation of multi-sensory signals.(ABSTRACT TRUNCATED AT 250 WORDS)


Attention and the subjective expansion of time.
During brief, dangerous events, such as car accidents and robberies, many people report that events seem to pass in slow motion, as if time had slowed down. We have measured a similar, although less dramatic, effect in response to unexpected, nonthreatening events. We attribute the subjective expansion of time to the engagement of attention and its influence on the amount of perceptual information processed. We term the effect time's subjective expansion (TSE) and examine here the objective temporal dynamics of these distortions. When a series of stimuli are shown in succession, the low-probability oddball stimulus in the series tends to last subjectively longer than the high-probability stimulus even when they last the same objective duration. In particular, (1) there is a latency of at least 120 msec between stimulus onset and the onset of TSE, which may be preceded by subjective temporal contraction; (2) there is a peak in TSE at which subjective time is particularly distorted at a latency of 225 msec after stimulus onset; and (3) the temporal dynamics of TSE are approximately the same in the visual and the auditory domains. Two control experiments (in which the methods of magnitude estimation and stimulus reproduction were used) replicated the temporal dynamics of TSE revealed by the method of constant stimuli, although the initial peak was not apparent with these methods. In addition, a third, control experiment (in which the method of single stimuli was used) showed that TSE in the visual domain can occur because of semantic novelty, rather than image novelty per se. Overall, the results support the view that attentional orienting underlies distortions in perceived duration.


Men's Vasectomy Knowledge, Attitudes, and Information-Seeking Behaviors in the Southern United States: Results From an Exploratory Survey.
Vasectomy is one of the few options men have to manage their reproductive capacity and take on a more equitable role in pregnancy prevention. While the method is underused throughout the United States, the southern states have a lower prevalence rate compared to the rest of the country. Existing survey research does not assess what men know or think about the procedure as a means of understanding why this is the case. We created and conducted an exploratory survey to assess men's knowledge, attitudes, and information-seeking behaviors about vasectomy in the Southern United States. We used targeted Facebook advertising to recruit men ages 25-70 years living in 7 southern states to complete an online survey (n = 397). Using regression analyses, we identify that participants who had a vasectomy knew more about the procedure than participants who had not. Participants who had not had a vasectomy had less positive attitudes about the procedure across all six attitude subscales compared to participants with vasectomies. We highlight potential avenues for future research to understand why this may be the case. Finally, the majority of participants knew someone who had had a vasectomy. This suggests that men disclose having a vasectomy to others. The interpersonal dynamics around vasectomy decision-making and disclosure remain unknown and a viable area for future research. Findings from this exploratory survey may be used by public health officials interested in implementing campaigns to increase knowledge about vasectomy and reduce stigma, which may encourage more positive attitudes about the procedure.


Archaeology and cognitive evolution.
Archaeology can provide two bodies of information relevant to the understanding of the evolution of human cognition--the timing of developments, and the evolutionary context of these developments. The challenge is methodological. Archaeology must document attributes that have direct implications for underlying cognitive mechanisms. One example of such a cognitive archaeology is found in spatial cognition. The archaeological record documents an evolutionary sequence that begins with ape-equivalent spatial abilities 2.5 million years ago and ends with the appearance of modern abilities in the still remote past of 400,000 years ago. The timing of these developments reveals two major episodes in the evolution in spatial ability, one, 1.5 million years ago and the other, one million years later. The two episodes of development in spatial cognition had very different evolutionary contexts. The first was associated with the shift to an open country adaptive niche that occurred early in the time range of Homo erectus. The second was associated with no clear adaptive shift, though it does appear to have coincided with the invasion of more hostile environments and the appearance of systematic hunting of large mammals. Neither, however, occurred in a context of modern hunting and gathering.


Safety of aprotinin in congenital heart operations: results from a large multicenter database.
Aprotinin was used frequently in children undergoing congenital heart operations with the aim of reducing bleeding, until it was taken off the market after adult studies reported increased renal failure and death. We evaluated the safety of aprotinin in a large multicenter cohort of children undergoing congenital heart operations. The Pediatric Health Information Systems Database was used to evaluate aprotinin in children (aged 0 to 18 years) undergoing congenital heart operations at 35 children's hospitals from 2003 to 2007. Propensity scores were constructed to account for potential confounders of age, sex, race, prematurity, genetic syndrome, type of operation (Risk Adjustment in Congenital Heart Surgery-1 category), center, and center volume. Multivariable analysis, adjusting for propensity score and individual covariates was performed to evaluate in-hospital mortality, postoperative renal failure requiring dialysis, and length of stay. Subanalysis was performed in the high-risk cohort undergoing reoperation. The study included 30,372 patients (56% male) who were a median age of 7 months (interquartile range, 36 days to 3.2 years). Overall, 44% received aprotinin. Multivariable analysis found no difference in postoperative mortality (odds ratio, 1.00; 95% confidence interval [CI], 0.99 to 1.01), dialysis (odds ratio, 1.00; 95% CI, 0.99 to 1.01), or length of stay (least square mean difference, -0.44 days; 95% CI, -1.01 to 0.13) between aprotinin recipients and nonrecipients. No difference was found in mortality or dialysis in patients undergoing reoperation. Aprotinin recipients in the reoperation subgroup had significantly reduced length of stay (least square mean difference, -2.05 days; 95% CI -3.29 to -0.81), These data suggest aprotinin is not associated with increased mortality or dialysis in children undergoing congenital heart operations and that further evaluation of aprotinin in this population could be undertaken without undue risk.


Capture of magnetic carriers within large arteries using external magnetic fields.
Our overall research goal is to advance the safety and effectiveness of acute ischemic stroke therapy by improving the benefit/risk ratio of thrombolysis and hence, the long-term outcome of acute ischemic stroke victims. Our approach is the development of a novel tissue plasminogen activator (t-PA) delivery system based on t-PA-loaded magnetic nano- and microcarriers guided directly to the site of vascular occlusion by external magnetic fields. Such a t-PA delivery system would conveniently combine the advantages of both intravenous (systemic) and intraarterial (catheter-facilitated) thrombolysis: non-invasiveness - the magnetic t-PA carriers can be injected intravenously and targeted, as drug delivery is magnetically guided to and t-PA focally released at and within the vascular clot to induce lysis. The focus of our discussion are the two necessary, fundamental and interrelated bioengineering steps: the research and development of well-characterized, biocompatible, functionally active and t-PA-loaded (encapsulated) magnetic nano- and microcarriers able to induce effective thrombolysis, and the design of magnetic guidance systems for targeted tPA-delivery allowing also the triggered release of the thrombolytic agent at the clot site. In this paper, we theoretically demonstrated magnetic trapping of blood borne magnetic nano- and microcarriers from human large vessels, especially arteries. Then, some preliminary experiments using primate models (monkeys) were done to identify successful in vivo sequestration of magnetic carriers in large and smaller arterial branches after arterial upstream and systemic venous injection. Histology (hematoxylin-eosin stain) verified intraarterial carrier concentration (identified as black carrier agglomerates on H and E staining) at the arterial region above the surface magnet. The results revealed the feasibility of magnetic drug-targeting at arteries and solidified the proposed t-PA delivery system.


Music Therapy Supports Children with Neurological Diseases during Physical Therapy Interventions.
Recent research found evidence supporting music therapy for children with neurological diseases during their hospitalized neurological early rehabilitation to promote their development during physical therapy. We hypothesized that live music therapy might improve vital signs during a physical therapy session. Seventeen children received live music therapy during the physical therapy session twice a week. Two more physical therapy sessions per week were held without music therapy. Heart rate, respiratory rate and oxygen saturation were recorded from 15 min before to 15 min after the therapy sessions. Physical therapy interventions showed changes in heart rate, respiratory rate and oxygen saturation between, before and after the sessions with or without music therapy. Live music therapy was effective for the vital signs during the intervention. We observed significantly lower heart and respiratory rates and higher oxygen saturation during physical therapy intervention with live music therapy in general (mean differences -8.0 beats per min; -0.8 breaths per min and +0.6%). When physical therapy was applied without music therapy children's heart rates increased by 8.5 beats per min and respiratory rates increased by 1.0 breaths per min. Live music therapy leads to a decrease in heart and respiratory rates and an increase in oxygen saturation in children with neurological diseases during physical therapy with live music therapy. Music therapy supports the children in physical therapy interventions during their hospitalization.


Preparing community pharmacy teams for health information exchange (HIE).
Health information exchanges (HIEs) facilitates health care professionals' electronic sharing of patient information across different organizations. When community pharmacists have access to HIE they can further contribute to improved patient outcomes. However, several implementation challenges are noted, which impede sustained pharmacist access to HIE. To our knowledge, no bidirectional HIE interface design and pharmacy team informed implementation process has been documented. In response, our research team designed and developed an HIE interface prototype for use specifically by community pharmacy teams to access local HIE data through their pharmacy dispensing software. To 1) identify barriers, facilitators, and recommendations for using HIE data in community pharmacies and 2) create a curated list of resources addressing identified implementation needs to aid future implementation of a fully functional, bidirectional HIE interface by community pharmacy teams. Pharmacists, pharmacy technicians, and patients from three pharmacy sites within the Community Pharmacy Enhanced Services Network of Indiana participated in individual semi-structured interviews. Interview questions were mapped to select constructs across all domains of the Consolidated Framework for Implementation Research. Interview transcripts were deductively coded. A subset of participants participated in Evidence Based Quality Improvement sessions to iteratively update planned resource items needed to support future HIE implementation. We interviewed 23 total participants: 8 pharmacists, 8 pharmacy technicians, and 7 patients. Five facilitators, four barriers, and two recommendations were identified. These were further characterized into four key implementation needs: instruction on how to use HIE; guidance on workflow and team roles; resources that are patient- facing; and resources that are provider-facing, resulting in 16 planned implementation resources. Our study provides the first-of-its-kind list of pharmacy team informed resources to facilitate sustainability and scalability of HIE implementation in community pharmacies.