Skin Delivery and Irritation Potential of Phenmetrazine as a Candidate Transdermal Formulation for Repurposed Indications.
Phenmetrazine, a selective dopamine and norepinephrine releaser, previously available as an oral anorectic, is prone to be abused. This study aimed to assess the feasibility of delivering phenmetrazine via the transdermal route for a new indication, while also minimizing its abuse potential. The passive permeation of phenmetrazine through dermatomed human cadaver skin was evaluated using static Franz diffusion cells at 10 mg/mL for the fumarate salt, and at 20, 40, and 80 mg/mL for the free base in propylene glycol for 24 h. Further, oleic acid (5% w/w), oleyl alcohol (5% and 10% w/w), and lauric acid (10% w/w) were investigated as chemical permeation enhancers to enhance the delivery. Skin irritation potential was assessed using EpiDerm™ in vitro reconstructed human epidermal model. The free base showed superior 24-h delivery (8.13 ± 4.07%, 10.6 ± 2.5%, and 10.4 ± 1.4% for groups with 20, 40, and 80 mg/mL of the free base, respectively) to phenmetrazine fumarate salt (undetectable). The successful screening of effective chemical enhancers, oleyl alcohol (5% and 10% w/w), oleic acid (5% w/w), and lauric acid (10% w/w) resulted in significant enhancement of delivery. The calculated therapeutic relevant flux for the potential indication, attention deficit hyperactivity disorder, 20 μg/cm2/h was met, where a 24-mg daily dose from a 50-cm2 patch was projected to be delivered to a 60-kg individual. Irritation study results suggest that formulations with therapeutically relevant delivery are likely to be non-irritant. In conclusion, it is feasible to deliver therapeutically relevant amounts of phenmetrazine via the transdermal route.


Nmnat delays axonal degeneration caused by mitochondrial and oxidative stress.
Axonal degeneration is a prominent feature of many neurological disorders that are associated with mitochondrial dysfunction, including Parkinson's disease, motor neuron disease, and inherited peripheral neuropathies. Studies of the Wld(s) mutant mouse, which undergoes delayed Wallerian degeneration in response to axonal injury, suggest that axonal degeneration is an active process. Wld(s) mice also have slower axonal degeneration and disease progression in numerous models of neurodegenerative disease. The Wld(s) mutation results in the production of a chimeric protein that contains the full-length coding sequence of nicotinamide mononucleotide adenylyltransferase 1 (Nmnat1), which alone is sufficient for axonal protection in vitro. To test the effects of increased Nmnat expression on axonal degeneration induced by mitochondrial dysfunction, we examined dorsal root ganglion (DRG) neurons treated with rotenone. Rotenone induced profound axonal degeneration in DRG neurons; however, this degeneration was delayed by expression of Nmnat. Nmnat-mediated protection resulted in decreased axonal accumulation and sensitivity to reactive oxygen species (ROS) but did not affect the change in the rate of rotenone-induced loss in neuronal ATP. Nmnat also prevented axonal degeneration caused by exposure to exogenous oxidants and reduced the level of axonal ROS after treatment with vincristine, further supporting the idea that Nmnat promotes axonal protection by mitigating the effects of ROS.


Identifying at-risk children at school entry: the usefulness of multibehavioral problem profiles.
Found that 1st-grade teacher ratings of aggressive, hyperactive-inattentive, and low levels of prosocial behaviors made unique contributions to the prediction of school outcomes (measured 2 years later) for 755 children. Person-oriented analyses compared the predictive utility of 5 screening strategies based on child problem profiles to identify children at risk for school problems. A broad screening strategy, in which children with elevations in any 1 of the 3 behavior problem dimensions were identified as "at-risk," showed lower specificity but superior sensitivity, odds ratios, and overall accuracy in the prediction of school outcomes than the other screening strategies that were more narrowly focused or were based on a total problem score. Results are discussed in terms of implications for the screening and design of preventive interventions.


Combination strategy for prognostication in patients undergoing post-resuscitation care after cardiac arrest.
This study investigated the prognostic performance of combination strategies using a multimodal approach in patients treated after cardiac arrest. Prospectively collected registry data were used for this retrospective analysis. Poor outcome was defined as a cerebral performance category of 3-5 at 6 months. Predictors of poor outcome were absence of ocular reflexes (PR/CR) without confounding factors, a highly malignant pattern on the most recent electroencephalography, defined as suppressed background with or without periodic discharges and burst-suppression, high neuron-specific enolase (NSE) after 48 h, and diffuse injury on imaging studies (computed tomography or diffusion-weighted imaging [DWI]) at 72-96 h. The prognostic performances for poor outcomes were analyzed for sensitivity and specificity. A total of 130 patients were included in the analysis. Of these, 68 (52.3%) patients had poor outcomes. The best prognostic performance was observed with the combination of absent PR/CR, high NSE, and diffuse injury on DWI [91.2%, 95% confidence interval (CI) 80.7-97.1], whereas the combination strategy of all available predictors did not improve prognostic performance (87.8%, 95% CI 73.8-95.9). Combining three of the predictors may improve prognostic performance and be more efficient than adding all tests indiscriminately, given limited medical resources.


Effect of chloride ions on the adsorption of 3-mercapto-1-propanesulfonic acid and bis(3-sulfopropyl)-disulfide on a Au(111) surface.
In situ scanning tunneling microscopy (STM) and cyclic voltammetry (CV) were used to study the adsorption of 3-mercapto-1-propanesulfonic acid (MPS) and bis(3-sulfopropyl)-disulfide (SPS) on Au(111) electrode in a HClO(4) aqueous solution. Chloride ions were introduced into the electrolyte solution, and their effect on the adsorption behavior of MPS and SPS was investigated. The CV results show that SPS and MPS molecules preferentially adsorb on the Au(111) surface compared to chloride ions, and furthermore, chloride ion can induce the adsorption of thiol molecules on the Au(111) surface. In the absence of chloride, no adsorption phase of SPS (or MPS) adlayer can be imaged by STM at low potentials. Raising electrode potential leads to the appearance of disordered adsorption phase at ca. 0.4 V (vs RHE) and ordered adlattices at ca. 0.8 V. In the presence of chloride, ordered adsorption structures of SPS and MPS appear at a lower potential (0.2 V), implying the enhancement effect of chloride to the thiol adsorption. It is inferred that the presence of chloride ions triggers a more positively charged gold surface, enhancing the reaction rate of thiol adsorption. Furthermore, the presence of chloride also leads to a decrease in the thiol-electrolyte interaction, due to the high solvation effect of chloride ions, which promotes the adsorption of SPS and MPS onto the Au surface. With further elevation of electrode potential, electrostatic interaction leads to coadsorption of chloride ions into the adlayer, as well as orientation changes of the ad-molecules. As a result, the ordered adlattice was disrupted and disappeared at ca. 0.5 V.


Runtime unknown unsafe scenarios identification for SOTIF of autonomous vehicles.
Safety is a critical concern for autonomous vehicles (AVs). Current testing approaches face challenges in simultaneously meeting the requirements of being valid, safe, and fast. To address these challenges, the silent testing approach that tests functions or systems in the background without interfering with driving is motivated. Building upon our previous research, this study first extends the method to specifically address the validation of AV perception, utilizing a lane marking detection algorithm (LMDA) as a case study. Second, field experiments were conducted to investigate the method's effectiveness in validating AV systems. For both studies, an architecture for describing the working principle is presented. The efficacy of the method in evaluating the LMDA is demonstrated through the use of adversarial images generated from a dataset. Furthermore, various scenarios involving pedestrians crossing a road under different levels of criticality were constructed to gain practical insights into the method's applicability for AV system validation. The results show that corner cases of the LMDA are successfully identified by the given evaluation metrics. Furthermore, the experiments highlight the benefits of employing multiple virtual instances with different initial states, enabling the expansion of the test space and the discovery of unknown unsafe scenarios, particularly those prone to false-positive objects. The practical implementation and systematic discussion of the method offer a significant contribution to AV safety validation.


Using the placebo response in clinical practice.
This paper reviews the psychological mechanisms that lead to placebo responses and the physiological basis for reduction in symptoms. Some psychological mechanisms (expectancy, conditioning) lead to symptom reduction but are unlikely to reduce underlying pathology. Other mechanisms (therapeutic relationship, empowerment) may additionally reduce later pathology. The nature, size and duration of the placebo response depends on the placebo inducing context. In clinical practice, the placebo response creates an adjunctive response to that of active treatment. It is a useful, but fickle, boon as it is difficult to predict when it will occur.


Clinical implications for breath-powered powder sumatriptan intranasal treatment.
The acute treatment of migraine requires matching patient need to drug and formulation. In particular, nausea and vomiting, quick time to peak intensity, and the common gastroparesis of migraineurs, all call for a variety of non-oral formulations for treatment of attacks. A novel breath-powered powder sumatriptan intranasal treatment offers an improvement, at least in pharmacokinetics, over conventional liquid nasal sumatriptan spray. The device for delivery in this breath-powered nasal sumatriptan uses natural nose anatomy to close the soft palate and propel the sumatriptan high up in the nasal cavity on one side with bidirectional airflow coming out the other side. This approach has the potential to reduce adverse events and improve efficacy. Phase 3 data on this system are in press at the time of this writing and results appear promising. The clinical role for a fast acting non-oral nasal formulation will be in those for whom tablets are bound to fail, that is, in the setting of nausea and vomiting or when the time to central sensitization, allodynia, and disabling migraine is too short for the patient to respond to a tablet. This review provides a clinical perspective on the breath-powered powder sumatriptan intranasal treatment.


Parmodel: a web server for automated comparative modeling of proteins.
Parmodel is a web server for automated comparative modeling and evaluation of protein structures. The aim of this tool is to help inexperienced users to perform modeling, assessment, visualization, and optimization of protein models as well as crystallographers to evaluate structures solved experimentally. It is subdivided in four modules: Parmodel Modeling, Parmodel Assessment, Parmodel Visualization, and Parmodel Optimization. The main module is the Parmodel Modeling that allows the building of several models for a same protein in a reduced time, through the distribution of modeling processes on a Beowulf cluster. Parmodel automates and integrates the main softwares used in comparative modeling as MODELLER, Whatcheck, Procheck, Raster3D, Molscript, and Gromacs. This web server is freely accessible at .


Simultaneous quantification of atomoxetine as well as its primary oxidative and O-glucuronide metabolites in human plasma and urine using liquid chromatography tandem mass spectrometry (LC/MS/MS).
A sensitive and selective liquid chromatography tandem mass spectrometry (LC/MS/MS) method for the determination of atomoxetine and its metabolites (4-hydroxyatomoxetine, N-des-methylatomoxetine, and 4-hydroxyatomoxetine-O-glucuronide) has been developed for human plasma and urine. Using stable-labeled internal standards, the method proved to be accurate and precise for the analytes in all species, resulting in inter-batch accuracy (percent relative error, %RE) within 100+/-13% and inter-batch precision (relative standard deviation, %RSD) within 11%. Stability was demonstrated for the analytes in neat solutions and the reconstitution solvent, as well as plasma and urine (with or without the deconjugation reagent). The method was simple, robust (utilized for the analysis of several hundred clinical study samples), and amenable to high sample throughput.


Pulp response after application of two resin modified glass ionomer cements (RMGICs) in deep cavities of prepared human teeth.
This study evaluated the human pulp response to the application of two RMGICs in deep cavities in vivo. The cavity floor prepared on the buccal surface of 34 premolars was lined with VBP (VBP), Vitrebond (VB) or Dycal® (DY), and restored with composite resin. Additional teeth were used as an intact control group. After 7 or 30-60 days, the teeth were extracted and processed for histological evaluation. The following histological events were scored: inflammatory response, tissue disorganization, reactionary dentin formation and presence of bacteria. At 7 days, VBP and VB elicited a mild inflammatory pulpal response in about 70% of specimens and in 1 specimen for DY. Only 1 specimen of each RMGICs exhibited moderate tissue disorganization. Bacteria and reactionary dentin formation were not found. At 30-60 days, about 20% of specimens lined with RMGICs showed a persistent mild inflammatory pulp response while no inflammatory reaction was observed for DY. Moderate tissue disorganization occurred with both materials. Bacteria were found only in 1 VBP specimen. The mean remaining dentin thickness (RDT) in specimens lined with VBP, VB or DY ranged from 342.3 to 436.1μm, and no statistically significant differences in RDT were found among materials or periods (two-way ANOVA, p>0.05). Comparison of the two RMGICs tested for the histological events at each period showed statistically similar results (Kruskal-Wallis, p>0.05). The use of the new Vitrebond formulation (VBP) in deep cavities in vivo caused mild initial pulp damage, which decreased with time, indicating acceptable biocompatibility.


Red cell indices: differentiation between β-thalassemia trait and iron deficiency anemia and application to sickle-cell disease and sickle-cell thalassemia.
In Tunisia, thalassemia and sickle cell disease represent the most prevalent monogenic hemoglobin disorders with 2.21% and 1.89% of carriers, respectively. This study aims to evaluate the diagnosis reliability of a series of red blood cell indices and parameters in differentiation of beta-thalassemia trait (β-TT) from iron deficiency anemia (IDA) and between homozygous sickle cell disease (SS) and sickle cell-thalassemia (ST). The study covered 384 patients divided into three groups. The first one is composed of 145 control group, the second consists of 57 β-TT and 52 IDA subjects and the last one with 88 SS and 42 ST patients. We calculated sensitivity, specificity, positive-predictive values, negative-predictive values, percentage of correctly identified patients and Youden's index for each indice. We also established new cut-off values by receiver operating characteristic curves for each indice. An evaluation study was performed on another population composed of 106 β-TT, 125 IDA, 31 SS and 17 ST patients. Srivastava Index, mean corpuscular hemoglobin, red blood cell, Mentzer Index (MI) and mean corpuscular hemoglobin concentration show the highest reliability in discriminating β-TT from IDA with new cut-offs slightly different from those described in literature. Ehsani Index, mean corpuscular volume, MI, Shine and Lal Index and Sirdah Index are the most powerful in the differentiation between SS and ST. The effectiveness and the simplicity of calculation of these indices make them acceptable and easy to use for differential diagnosis.


Processing Non-at-Issue Meanings of Conditional Connectives: The wenn/falls Contrast in German.
Logical connectives in natural language pose challenges to truth-conditional semantics due to pragmatics and gradience in their meaning. This paper reports on a case study of the conditional connectives (CCs) wenn/falls 'if/when, if/in case' in German. Using distributional evidence, I argue that wenn and falls differ in lexical pragmatics: They express different degrees of speaker commitment (i.e., credence) toward the modified antecedent proposition at the non-at-issue dimension. This contrast can be modeled using the speaker commitment scale (Giannakidou and Mari, 2016), i.e., More committed<WENN p, FALLS p>Less committed. Four experiments are reported which tested the wenn/falls contrast, as well as the summary of an additional one from Liu (2019). Experiment 1 tested the naturalness of sentences containing the CCs (wenn or falls) and conditional antecedents with varying degrees of likelihood (very likely/likely/unlikely). The starting prediction was that falls might be degraded in combination with very likely and likely events in comparison to the other conditions, which was not borne out. Experiment 2 used the forced lexical choice paradigm, testing the choice between wenn and falls in the doxastic agent's conditional thought, depending on their belief or disbelief in the antecedent. The finding was that subjects chose falls significantly more often than wenn in the disbelief-context, and vice versa in the belief-context. Experiment 3 tested the naturalness of sentences with CCs and an additional relative clause conveying the speaker's belief or disbelief in the antecedent. An interaction was found: While in the belief-context, wenn was rated more natural than falls, the reverse pattern was found in the disbelief-context. While the results are mixed, the combination of the findings in Experiment 2, Experiment 3 and that of Experiment 4a from Liu (2019) that falls led to lower speaker commitment ratings than wenn, provide evidence for the CC scale. Experiment 4b tested the interaction between two speaker commitment scales, namely, one of connectives (including weil 'because' and wenn/falls) and the other of adverbs (factive vs. non-factive, Liu, 2012). While factive and non-factive adverbs were rated equally natural for the factive causal connective, non-factive adverbs were preferred over factive ones by both CCs, with no difference between wenn and falls. This is discussed together with the result in Liu (2019), where the wenn/falls difference occurred in the absence of negative polarity items (NPIs), but disappeared in the presence of NPIs. This raises further questions on how different speaker commitment scales interact and why.


New Classification of Neisseria meningitidis by Means of Bactericidal Reactions.
A bactericidal assay is described which allows identification of distinct serotypes within a serogroup of Neisseria meningitidis. Antisera produced in rabbits against seven group C strains by two intravenous inoculations of live organisms were found to contain two types of bactericidal antibodies. One, directed against the group-specific polysaccharide, caused various degrees of killing of all strains. Absorption of this antibody by purified group C polysaccharide revealed the presence of the second bactericidal antibody. This antibody was directed against antigenically distinct factors associated with serotype specificity. Extensive cross-absorption yielded antisera with activity directed against four separate factors. The presence of a factor in a strain was indicated by its susceptibility to killing by antisera containing antibody to that factor. A serotype was defined by the particular combination of factors. Six different serotypes, containing one or two factors, were identified among 16 group C strains examined.


Long-term evaluation of mucosal and systemic immunity and protection conferred by different polio booster vaccines.
Oral polio vaccine (OPV) and Inactivated Polio Vaccine (IPV) have distinct advantages and limitations. IPV does not provide mucosal immunity and introduction of IPV to mitigate consequences of circulating vaccine-derived polio virus from OPV has very limited effect on transmission and OPV campaigns are essential for interrupting wild polio virus transmission, even in developed countries with a high coverage of IPV and protected sewer systems. The problem is magnified in many countries with limited resources. Requirement of refrigeration for storage and transportation for both IPV and OPV is also a major challenge in developing countries. Therefore, we present here long-term studies on comparison of a plant-based booster vaccine, which is free of virus and cold chain with IPV boosters and provide data on mucosal and systemic immunity and protection conferred by neutralizing antibodies. Mice were primed subcutaneously with IPV and boosted orally with lyophilized plant cells containing 1μg or 25μg polio viral protein 1 (VP1), once a month for three months or a single booster one year after the first prime. Our results show that VP1-IgG1 titers in single or double dose IPV dropped to background levels after one year of immunization. This decrease correlated with >50% reduction in seropositivity in double dose and <10% seropositivity in single dose IPV against serotype 1. Single dose IPV offered no or minimal protection against serotype 1 and 2 but conferred protection against serotype 3. VP1-IgA titers were negligible in IPV single or double dose vaccinated mice. VP1 antigen with two plant-derived adjuvants induced significantly high level and long lasting VP1-IgG1, IgA and neutralizing antibody titers (average 4.3-6.8 log2 titers). Plant boosters with VP1 and plant derived adjuvants maintained the same level titers from 29 to 400days and conferred the same level of protection against all three serotypes throughout the duration of this study. Even during period, when no plant booster was given (∼260days), VP1-IgG1 titers were maintained at high levels. Lyophilized plant cells expressing VP1 can be stored without losing efficacy, eliminating cold chain. Virus-free, cold-chain free vaccine is ready for further clinical development.


Pancreatic abnormalities and AIDS related sclerosing cholangitis.
Biliary tract abnormalities are well recognised in AIDS, most frequently related to opportunistic infection with Cryptosporidium, Microsporidium, and cytomegalovirus. We noted a high frequency of pancreatic abnormalities associated with biliary tract disease. To define these further we reviewed the clinical and radiological features in these patients. Notes and radiographs were available from two centres for 83 HIV positive patients who had undergone endoscopic retrograde cholangiopancreatography for the investigation of cholestatic liver function tests or abdominal pain. 56 patients had AIDS related sclerosing cholangitis (ARSC); 86% of these patients had epigastric or right upper quadrant pain and 52% had hepatomegaly. Of the patients with ARSC, 10 had papillary stenosis alone, 11 had intra- and extrahepatic sclerosing cholangitis alone, and 35 had a combination of the two. Ampullary biopsies performed in 24 patients confirmed an opportunistic infection in 16. In 15 patients, intraluminal polyps were noted on the cholangiogram. Pancreatograms were available in 34 of the 45 patients with papillary stenosis, in which 29 (81%) had associated pancreatic duct dilatation, often with associated features of chronic pancreatitis. In the remaining 27 patients, final diagnoses included drug induced liver disease, acalculous cholecystitis, gall bladder empyema, chronic B virus hepatitis, and alcoholic liver disease. Pancreatic abnormalities are commonly seen with ARSC and may be responsible for some of the pain not relieved by biliary sphincterotomy. The most frequent radiographic biliary abnormality is papillary stenosis combined with ductal sclerosis.


Low-frequency ultrasound induces apoptosis of rat aortic smooth muscle cells (A7r5) via the intrinsic apoptotic pathway.
Ultrasound, a non-invasive therapy method, is a potential tool for medical applications, but its biological effects on vascular smooth muscle cells (VSMCs) have not been characterized. The aim of this study was to explore the effect and possible apoptotic mechanism of VSMCs that were induced by low-frequency ultrasound (LFU). Cell viability and apoptosis of A7r5 cells were evaluated after treating A7r5 cells with a continuous 45-kHz 1.0-W/cm(2) ultrasound (exposure time of 0, 10, 20, 30, and 35 s) by MTT assay and flow cytometry. At the optimum ultrasound exposure condition (30 s), gene chip analysis was performed, and the apoptotic signaling pathway was confirmed by reverse transcription-polymerase chain reaction and Western blot. As measured by flow cytometry, LFU significantly induced A7r5 cell apoptosis. Comparing the ultrasound group with the control group, the protein expression of caspase-9 and caspase-3 was increased by 50 and 57%, respectively; the caspase-3 mRNA level was increased by 37.5%. These findings indicate that an intrinsic pathway plays a major role in apoptosis that is induced by LFU and that LFU can induce A7r5 cell apoptosis via caspase-9- and caspase-3-dependent pathways.


Attitudes toward dental appearance in 50- and 60-Year-old subjects living in Sweden.
Attitudes toward the importance of general appearance have varied much, not only over time but also among groups of individuals of different age and other characteristics. Whether the situation is similar regarding dental appearance does not seem to have been studied. The aim was to study the differences in attitudes toward dental appearance between two large samples of 50- and 60-year-old subjects. Identical questionnaires were sent to all subjects born in 1942 and 1952 living in two Swedish counties in 2002 (N=17,444; N50=8,881, N60=8,563). The final response rate was 72.2% (N=12,599). In this study, responses to four statements on the importance of dental appearance have been analyzed with respect to gender and age. Many of the responses to the four statements differed with gender and age. To the first statement ("To have beautiful and perfect teeth is very important for how you are treated by other people"), 73% agreed at age 60 compared with 64% at age 50 (p < 0.001). Approximately 90% of the subjects agreed with the second statement ("Minor esthetic imperfections of the teeth have no importance, only they function well"). Logistic regression indicated that several variables were significantly associated with the statements. Besides age and gender, education and self-assessed dental problems were most important. Attitudes toward the importance of dental appearance differed both between genders and age groups in these population samples living in Sweden. The varying attitudes toward dental appearance in the population must be acknowledged in treatment decisions.


Repositioning the Sm-Binding Site in Saccharomyces cerevisiae Telomerase RNA Reveals RNP Organizational Flexibility and Sm-Directed 3'-End Formation.
Telomerase RNA contains a template for synthesizing telomeric DNA and has been proposed to act as a flexible scaffold for holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae, the telomerase RNA, TLC1, is bound by the Sm7 protein complex, which is required for stabilization of the predominant, non-polyadenylated (poly(A)-) TLC1 isoform. However, it remains unclear (1) whether Sm7 retains this function when its binding site is repositioned within TLC1, as has been shown for other TLC1-binding telomerase subunits, and (2) how Sm7 stabilizes poly(A)- TLC1. Here, we first show that Sm7 can stabilize poly(A)- TLC1 even when its binding site is repositioned via circular permutation to several different positions within TLC1, further supporting the conclusion that the telomerase holoenzyme is organizationally flexible. Next, we show that when an Sm site is inserted 5' of its native position and the native site is mutated, Sm7 stabilizes shorter forms of poly(A)- TLC1 in a manner corresponding to how far upstream the new site was inserted, providing strong evidence that Sm7 binding to TLC1 controls where the mature poly(A)- 3' is formed by directing a 3'-to-5' processing mechanism. In summary, our results show that Sm7 and the 3' end of yeast telomerase RNA comprise an organizationally flexible module within the telomerase RNP and provide insights into the mechanistic role of Sm7 in telomerase RNA biogenesis.


The connection between illness representations of Alzheimer's disease and burnout among social workers and nurses in nursing homes and hospitals: a mixed-methods investigation.
To examine the relationship between Alzheimer's disease (AD) illness representations and burnout among social workers and nurses, based on the self-regulatory model. A mixed-methods study was conducted. First, 327 social workers and nurses completed measures of cognitive and emotional representations, burnout (emotional exhaustion, depersonalization, lack of personal accomplishment), role variables, knowledge about AD, emotion-focused coping, problem-focused coping, and demographic and occupational characteristics. Second, interviews were conducted with eight social workers and nurses to uncover their perceptions about AD and their burnout experience. Using structural equation modeling and controlling background variables, findings indicated that emotional representations were associated with burnout while only some of the cognitive illness representations were associated with burnout. While cognitive illness representations were associated directly to burnout, the association between emotional representations and burnout was mediated by emotion-focused coping. The trimmed model showed a good fit of the data and explained 32.2% of the variance in emotion-focused coping, 51% of the variance in emotional exhaustion, 37.7% of the variance in depersonalization, and 22.6% of the variance in lack of personal accomplishment. Interviews demonstrated that AD characteristics were perceived as affecting participants on both personal and professional levels; the participants expressed negative feelings towards AD and stated that these perceptions and feelings had led them to burnout. AD illness representations may be a risk factor for developing burnout. New directions for intervention programs, aiming to reduce burnout, should be examined.


Formulation and Evaluation of in situ Gels Containing Clotrimazole for Oral Candidiasis.
Gel dosage forms are successfully used as drug delivery systems to control drug release and protect the medicaments from a hostile environment. The main objective is to formulate and evaluate in situ oral topical gels of clotrimazole based on the concept of pH triggered and ion activated systems. The system utilizes polymers that exhibit sol-to-gel phase transition due to change in specific physico-chemical parameters. A pH triggered system consisting of carbopol 934P (0.2-1.4% w/v) and ion triggered system using gellan gum (0.1-0.75% w/v) along with hydroxylpropylmethylcelluose E50LV was used to prolong the release of clotrimazole (0.1% w/v). Formulations were evaluated for gelling capacity, viscosity, gel strength, bioadhesive force, spreadability, microbiological studies and in vitro release. The use of carbopol as in situ gel forming system was substantiated by the property to transform into stiff gels when the pH was raised, whereas in gellan gum this transformation occurred in the presence of monovalent/divalent cations. Effect of calcium carbonate and other process parameters optimized and found that increase in calcium ions produced stronger gels. The drug content, clarity, and pH of the formulation were found to be satisfactory. The viscosity was found to be in the range 5 to 85 centipoise for the sol, whereas for the gels it was up to 16000 centipoise. The formulation showed pseudoplastic flow with thixotrophy. The maximum gel strength (using texture analyzer) and bioadhesion was found to be up to 6.5 g and 4 g, respectively. The optimized formulations were able to release the drug up to 6 h. The formulation containing gellan gum showed better sustained release compared to carbopol based gels.


A novel electrochemical sensor for the analysis of β-agonists: the poly(acid chrome blue K)/graphene oxide-nafion/glassy carbon electrode.
A novel modified electrode was constructed by the electro-polymerization of 4,5-dihydroxy-3-[(2-hydroxy-5-sulfophenyl)azo]-2,7-naphthalenedisulfonic acid trisodium salt (acid chrome blue K (ACBK)) at a graphene oxide (GO)-nafion modified glassy carbon electrode (GCE). The characterization of an electrochemically synthesized poly-ACBK/GO-nafion film was investigated by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), atomic force microscopy (AFM) and scanning electron microscopy (SEM) techniques, and the results were interpreted and compared at each stage of the electrode construction. Electrochemical oxidation of eight β-agonists - clenbuterol, salbutamol, terbutaline, ractopamine, dopamine, dobutamine, adrenaline, and isoprenaline, was investigated by CV at the different electrodes. At the poly-ACBK/GO-nafion/GCE, the linear sweep voltammetry peak currents of the eight β-agonists increased linearly with their concentrations in the range of 1.0-36.0 ng mL(-1), respectively, and their corresponding limits of detection (LODs) were within the 0.58-1.46 ng mL(-1) range. This electrode showed satisfactory reproducibility and stability, and was used successfully for the quantitative analysis of clenbuterol in pork samples.


Hormone-Dependent Prostate Cancers are Dependent on Rac Signaling for Growth and Survival.
Prostate cancer remains a common cause of cancer mortality in men. Initially, cancers are dependent of androgens for growth and survival. First line therapies reduce levels of circulating androgens or target the androgen receptor (AR) directly. Although most patients show durable responses, many patients eventually progress to castration-resistant prostate cancer (CRPC) creating a need for alternative treatment options. The Rac1 signaling pathway has previously been implicated as a driver of cancer initiation and disease progression. We investigated the role of HACE1, the E3 ubiquitin ligase for Rac1, in prostate cancer and found that HACE1 is commonly lost resulting in hyperactive Rac signaling leading to enhanced cellular proliferation, motility and viability. Importantly, we show that a Rac inhibitor can attenuate the growth and survival of prostate cancer cells. Rac signaling was also found to be critical in prostate cancers that express the AR. Rac inhibition in androgen dependent cells resulted in reduction of AR target gene expression suggesting that targeting Rac1 may be an alternative method for blocking the AR signaling axis. Finally, when used in combination with AR antagonists, Rac inhibition enhanced the suppression of AR target gene expression. Therefore, targeting Rac in prostate cancer has the potential to enhance the efficacy of approved AR therapies.


Partial characterization of antigens from M. leprae evoking IgG and IgM antibodies in armadillos.
Armadillo IgG and IgM antibody responses to Mycobacterium leprae were analyzed using isotypic-specific antisera by means of immunoblotting. Blots developed for IgG antibodies to M. leprae showed multiple protein antigens (Mr = 12-90 K) in some heavily infected armadillos. In contrast, blots developed for IgM antibodies to M. leprae showed a single, broad, diffuse band of immunoreactivity at approximately 33 kDa. The 33-kDa immunogen was detectable with silver stain modified for carbohydrate reactivity, suggesting the presence of a polysaccharide component. In addition, binding of 125I-concanavalin A to the 33-kDa component demonstrated the presence of mannose and/or glucose residues.


Incidence and Clinical Significance of Acute Reocclusion after Emergent Angioplasty or Stenting for Underlying Intracranial Stenosis in Patients with Acute Stroke.
A major concern after emergent intracranial angioplasty in cases of acute stroke with underlying intracranial stenosis is the acute reocclusion of the treated arteries. This study reports the incidence and clinical outcomes of acute reocclusion of arteries following emergent intracranial angioplasty with or without stent placement for the management of patients with acute stroke with underlying intracranial atherosclerotic stenosis. Forty-six patients with acute stroke received emergent intracranial angioplasty with or without stent placement for intracranial atherosclerotic stenosis and underwent follow-up head CTA. Acute reocclusion was defined as "hypoattenuation" within an arterial segment with discrete discontinuation of the arterial contrast column, both proximal and distal to the hypoattenuated lesion, on CTA performed before discharge. Angioplasty was defined as "suboptimal" if a residual stenosis of ≥50% was detected on the postprocedural angiography. Clinical and radiologic data of patients with and without reocclusion were compared. Of the 46 patients, 29 and 17 underwent angioplasty with and without stent placement, respectively. Acute reocclusion was observed in 6 patients (13%) and was more frequent among those with suboptimal angioplasty than among those without it (71.4% versus 2.6%, P < .001). The relative risk of acute reocclusion in patients with suboptimal angioplasty was 27.857 (95% confidence interval, 3.806-203.911). Furthermore, a good outcome was significantly less frequent in patients with acute reocclusion than in those without it (16.7% versus 67.5%, P = .028). Acute reocclusion of treated arteries was common after emergent intracranial angioplasty with or without stent placement in patients with acute stroke with intracranial atherosclerotic stenosis and was associated with a poor outcome. Suboptimal results of angioplasty appear to be associated with acute reocclusion, irrespective of whether stent placement was performed.


Monoclonal antibodies to human vitamin D-binding protein.
Monoclonal antibodies to vitamin D-binding protein isolated from human serum have been produced. The antibodies obtained have been shown to be specific for human vitamin D-binding protein by three independent assays. The antibodies recognize human vitamin D-binding protein specifically in an enzyme-linked immunosorbent assay. Human vitamin D-binding protein is detected specifically in both pure and crude samples by a radiometric immunosorbent assay (RISA) and by an immunoprecipitation assay. The anti-human vitamin D-binding protein antibodies cross-react with monkey and pig vitamin D-binding protein, but not with vitamin D-binding protein from rat, mouse, or chicken, as determined by the RISA and immunoprecipitation assays.


The status of combination therapy for visceral leishmaniasis: an updated review.
For the past 15 years, trials of combination therapy options for visceral leishmaniasis have been conducted with the aim of identifying effective, and safe treatment regimens that were shorter than existing monotherapy regimens and could also prevent or delay the emergence of drug resistance. Although first-line treatment currently relies on combination therapy in east Africa, this is not true in Latin America owing to disappointing trial results, with lower than expected efficacy seen for the combination treatment group. By contrast, several effective combination therapy regimens have been identified through trials on the Indian subcontinent; yet, first-line therapy is still AmBisome monotherapy as the drug is part of a free donation programme and is highly effective in this region. Achieving a short all-oral combination treatment will require new chemical entities, several of which are currently under evaluation. Future studies should systematically include pharmacological substudies to ensure optimal dosing for all patient groups. To achieve maximal impact of new combination treatments, mechanisms to ensure drug availability and access after trials should be established. Enhancing the longevity of current and novel treatments will require effective systems for early detection of emerging drug resistance.


Differential requirements for clathrin endocytic pathway components in cellular entry by Ebola and Marburg glycoprotein pseudovirions.
Clathrin-mediated endocytosis was previously implicated as one of the cellular pathways involved in filoviral glycoprotein mediated viral entry into target cells. Here we have further dissected the requirements for different components of this pathway in Ebola versus Marburg virus glycoprotein (GP) mediated viral infection. Although a number of these components were involved in both cases; Ebola GP-dependent viral entry specifically required the cargo recognition proteins Eps15 and DAB2 as well as the clathrin adaptor protein AP-2. In contrast, Marburg GP-mediated infection was independent of these three proteins and instead required beta-arrestin 1 (ARRB1). These findings have revealed an unexpected difference between the clathrin pathway requirements for Ebola GP versus Marburg GP pseudovirion infection. Anthrax toxin also uses a clathrin-, and ARRB1-dependent pathway for cellular entry, indicating that the mechanism used by Marburg GP pseudovirions may be more generally important for pathogen entry.


Electrophysiological properties of rat mesenteric lymphatic vessels and their regulation by stretch.
In mammals, lymph is propelled centrally primarily via the phasic contractions of collecting lymphatic vessels, known as lymphatic pumping. Electrophysiological studies conducted in guinea pig and sheep mesenteric lymphatic vessels indicate that contractions are initiated in the lymphatic muscle by nifedipine-sensitive action potentials (APs). Lymphatic pumping is highly sensitive to luminal fluid loading and the mechanical properties of this stretch-induced pumping have been consistently studied, in particular in rat mesenteric lymphatic vessels. However, membrane potential (Vm) and the electrophysiological events underlying stretch-induced lymphatic pumping have not been investigated in the rat. The aim of this study was thus to examine the properties of rat mesenteric lymphatic muscle Vm under resting conditions and to assess changes in Vm caused by distension. Lymphatic muscle Vm was measured with sharp intracellular microelectrodes either in unstretched conditions or under isometric tension provided by a wire-myograph. In unstretched vessels, Vm was -48 ± 2 mV (n=30). APs (amplitude ∼25 mV) were observed at a frequency of ∼8/min and were abolished by nifedipine. Under isometric tension, Vm was less polarized (-36 ± 1 mV, n=23), even at minimum tension. Increase in tension led to increase in contraction strength and contraction/AP frequency, while Vm was slightly hyperpolarized and AP amplitude not markedly altered. In our experimental conditions, rat lymphatic muscle has electrophysiological characteristics similar to that in other species. It responds to an increase in isometric tension with an increase in AP frequency, but resting Vm is not significantly affected.


Functional development of the visual system in normal and protein deprived rats. II. Morphometric and biochemical studies on adult optic nerve.
The optic nerve of normal (C) and protein deprived (PD) adult rats was examined by morphometry and biochemistry. The mean cross-sectional area of the optic nerve was reduced by 15% and the number of axons per unit area increased by 17% in the PD rats. Calibre spectrum analysis of axons revealed a reduction in median diameter from 0.49 micron in controls to 0.45 micron in PD rats. The number of axons with a diameter larger than 1 micron was reduced by 35% in PD rats. These reductions were probably due to a general reduction in size, since the calculated total number of axons in the optic nerve was almost identical in C and PD rats (126 X 10(3) and 124 X 10(3), respectively). The increased packing density of axons in the nerve was not only due to thinner axons. The biochemical measurements showed a marked reduction in myelin basic protein in the optic nerves of PD rats, without an alteration in the composition of the total protein. This confirms the persistent hypomyelination which has been reported previously in other malnutrition models. The possible relations between the structural and biochemical changes affecting optic nerve fibres and physiological findings on cortical visual evoked response and on optic nerve in vitro in PD rats are discussed.


Chimeric nature of pinopsin between rod and cone visual pigments.
Chicken pineal pinopsin is the first example of extra-retinal opsins, but little is known about its molecular properties as compared with retinal rod and cone opsins. For characterization of extra-retinal photon signaling, we have developed an overexpression system providing a sufficient amount of purified pinopsin. The recombinant pinopsin, together with similarly prepared chicken rhodopsin and green-sensitive cone pigment, was subjected to photochemical and biochemical analyses by using low-temperature spectroscopy and the transducin activation assay. At liquid nitrogen temperature (-196 degrees C), we detected two kinds of photoproducts, bathopinopsin and isopinopsin, having their absorption maxima (lambda(max)) at 527 and approximately 440 nm, respectively, and we observed complete photoreversibility among pinopsin, bathopinopsin, and isopinopsin. A close parallel of the photoreversibility to the rhodopsin system strongly suggests that light absorbed by pinopsin triggers the initial event of cis-trans isomerization of the 11-cis-retinylidene chromophore. Upon warming, bathopinopsin decayed through a series of photobleaching intermediates: lumipinopsin (lambda(max) 461 nm), metapinopsin I (460 nm), metapinopsin II (385 nm), and metapinopsin III (460 nm). Biochemical and kinetic analyses showed that metapinopsin II is a physiologically important photoproduct activating transducin. Detailed kinetic analyses revealed that the formation of metapinopsin II is as fast as that of a chicken cone pigment, green, but that the decay process of metapinopsin II is as slow as that of the rod pigment, rhodopsin. These results indicate that pinopsin is a new type of pigment with a chimeric nature between rod and cone visual pigments in terms of the thermal behaviors of the meta II intermediate. Such a long-lived active state of pinopsin may play a role in the pineal-specific phototransduction process.


What is important to continuity in home care?. Perspectives of key stakeholders.
In Canada, home care is growing rapidly. Each province takes a somewhat different approach to its delivery. Ontario uses a competitive bidding model to award contracts to community agencies that bid for service delivery rights. Contracts are to be awarded based on quality and price. However, the attributes thought to contribute to high quality, such as continuity of care, are not clearly defined and are not measured. We sought to identify factors that were important to experiencing continuity of care in home care. We interviewed home care clients and their caregivers, workers in the home care system (nursing and homemaking service providers, case managers) and physicians whose patients use home care. During in-depth interviews with these key stakeholders, they described the conditions that led to continuity of care in home care. Service providers and case managers were also asked about the types of clients who need a high level of care continuity. Care that is experienced as running smoothly, that responds to clients' needs and requires no special effort for clients to maintain, was seen as having continuity. The attributes of care experienced as facilitating continuity could be grouped under two dimensions of care-managing care (care planning, monitoring and review; and care coordination) and direct service provision (uninterrupted service delivery; consistent, appropriate knowledge and skills; ongoing accurate observation; trusting relationship between service provider and client/caregiver; rapport among team members; and consistent timing). Different stakeholders emphasized different attributes of care as most important to continuity. Clients included consistency of timing of service delivery while rarely mentioning care management issues. They emphasized the importance of consistent knowledge and skills in the workers and trusting relationships as important to experiencing care continuity. The description of attributes of continuity of home care that emerged from this study is compared to definitions found in the nursing, mental health and primary care literature.


Incorporating Field Studies into Species Distribution and Climate Change Modelling: A Case Study of the Koomal Trichosurus vulpecula hypoleucus (Phalangeridae).
Species distribution models (SDMs) are an effective way of predicting the potential distribution of species and their response to environmental change. Most SDMs apply presence data to a relatively generic set of predictive variables such as climate. However, this weakens the modelling process by overlooking the responses to more cryptic predictive variables. In this paper we demonstrate a means by which data gathered from an intensive animal trapping study can be used to enhance SDMs by combining field data with bioclimatic modelling techniques to determine the future potential distribution for the koomal (Trichosurus vulpecula hypoleucus). The koomal is a geographically isolated subspecies of the common brushtail possum, endemic to south-western Australia. Since European settlement this taxon has undergone a significant reduction in distribution due to its vulnerability to habitat fragmentation, introduced predators and tree/shrub dieback caused by a virulent group of plant pathogens of the genus Phytophthora. An intensive field study found: 1) the home range for the koomal rarely exceeded 1 km in in length at its widest point; 2) areas heavily infested with dieback were not occupied; 3) gap crossing between patches (>400 m) was common behaviour; 4) koomal presence was linked to the extent of suitable vegetation; and 5) where the needs of koomal were met, populations in fragments were demographically similar to those found in contiguous landscapes. We used this information to resolve a more accurate SDM for the koomal than that created from bioclimatic data alone. Specifically, we refined spatial coverages of remnant vegetation and dieback, to develop a set of variables that we combined with selected bioclimatic variables to construct models. We conclude that the utility value of an SDM can be enhanced and given greater resolution by identifying variables that reflect observed, species-specific responses to landscape parameters and incorporating these responses into the model.


Morbidity, outcomes and cost-benefit analysis of wildlife rehabilitation in Catalonia (Spain).
There are few studies of careful examination of wildlife casualties in Wildlife Rehabilitation Centers. These studies are essential for detecting menaces to wild species and providing objective criteria about cost-benefit of treatments in those centers. The release rate is considered the main outcome indicator, but other parameters such as length of stay at the center and a cost-benefit index expressed as number of released animals per euro and day, could be used as reliable estimators of the rehabilitation costs. A retrospective study based on 54772 admissions recorded from 1995-2013 in the database of the Wildlife Rehabilitation Center of Torreferrussa (Catalonia, NW Spain) assessed the morbidity, outcomes and cost-benefits of the rehabilitation practices. Three hundred and two species were included: 232 birds (n = 48633), 37 mammals (n = 3293), 20 reptiles (n = 2705) and 13 amphibians (n = 141). The most frequent causes of admission were: 39.8% confiscation of protected species (89.4% passerines), 31.8% orphaned young animals (35.3% swifts, 21.7% diurnal raptors and owls) and 17.4% trauma casualties (46.7% raptors and owls). The highest proportion of releases was found in the captivity confiscation category [87.4% passerines (median time of stay: 12 days)], followed by the orphaned category [78% owls (66 days), 76.5% diurnal birds of prey (43 days), 75.6% hedgehogs (49 days), 52.7% swifts (19 days) and 52% bats (55 days)]. For the trauma group, 46.8% of releases were hedgehogs (44 days) and 25.6% owls (103 days). As regards the cost-benefit index, the trauma casualties and infectious diseases had the worse values with 1.3 and 1.4 released animals/euro/day respectively, and were particularly low in raptors, waders, marine birds and chiroptera. On the contrary, captivity (4.6) and misplacement (4.1) had the best index, particulary in amphibian, reptiles and passerines. Cost-benefit studies including the release rate, the time of stay at the center and the cost-benefit index should be implemented for improving management efficiency of the Wildlife Rehabilitation Centers.


Subcellular distribution and movement of 5'-nucleotidase in rat cells.
1. Cell-surface 5'-nucleotidase was assayed by incubating whole-cell suspensions with 5'[3H]-AMP in iso-osmotic buffer and measuring [3H]adenosine production. The activity of cell-surface 5'-nucleotidase in hepatocytes, adipocytes and lymphocytes isolated from the rat was 15.0, 0.5 and 0.8pmol/min per cell at 37 degrees C respectively. 2. Disruption of the cells by vigorous mechanical homogenization or detergent treatment exposed additional 5'-nucleotidase activity, which represented 52%, 25% and 21% of the total activity in the three cell types respectively. This increase in 5'-nucleotidase activity which occurred when the cells were homogenized was due to a second pool of 5'-nucleotidase within the cell, rather than activation of the cell-surface enzyme. 3. In hepatocytes the intracellular 5'-nucleotidase activity was membrane-bound, indistinguishable from cell-surface 5'-nucleotidase in its inhibition by rabbit anti-(rat liver 5'-nucleotidase) serum and its kinetics with AMP, and was located on the extracytoplasmic face of vesicles within the cell. 4. The cell-surface 5'-nucleotidase of rat hepatocytes was rapidly inhibited when rabbit anti-(rat liver 5'-nucleotidase) serum or concanavalin A was added to the medium at 37 degrees C. Incubation with antiserum for 5 min at 37 degrees C inhibited 83 +/- 3% of the cell-surface enzyme. 5. Incubation of hepatocytes with exogenous antiserum or concanavalin A for 30 min at 37 degrees C resulted in over 50% inhibition of the intracellular enzyme. This inhibition was not prevented by disruption of the cytoskeleton or by ATP depletion. 6. Incubation of hepatocytes with exogenous antiserum or concanavalin A for up to 2h at 0 degrees C caused little or no inhibition of the intracellular enzyme, but over 75% inhibition of the cell-surface enzyme. 7. When surface-inhibited hepatocytes were washed and resuspended in buffer at 37 degrees C, 5'-nucleotidase was observed to redistribute from the intracellular pool to the cell surface.


Relation of lichen sclerosus et atrophicus of the vulva to development of carcinoma.
One hundred and seven patients with lichen sclerosus et atrophicus (LS&A) of the vulva were studies to determine the malignant potential of the LS&A. Five patients had coexisting invasive carcinoma of the vulva or perineum with the LS&A, and 1 patient had coexisting intraepithelial vulvar carcinoma on the clitoris. None of these, however, was known to have LS&A prior to the biopsy for carcinoma of the vulva. The high association of carcinoma and LS*A is probably a result of selection of 2 unusual lesions sent for consultation and evaluation. Squamous hyperplasia in the vulva occurred in association with LS&A in 37 (35%) patients, but only 6 patients had areas of dysplasia coexisting with LS&A. These areas of dysplasia, like the 5 invasive carcinomas, occurred in an area of the vulva where the LS&A was minimal or absent. Follow-up data were obtained on 92 patients with LS&A. Only 1 developed carcinoma of the vulva, which occurred 12 years after identification of the LS&A. When carcinoma arises in the vulva in a patient with vulvar LS&A, it tends to arise in areas of minomal LS&A or isolated areas of relatively normal vulvar skin. This study did not provide evidence of carcinoma arising from LS&A. Five of the 92 patients developed 6 malignant neoplasms in other sites, including carcinoma of the endometrium (3 patients), lung (1 patient), and simultaneous carcinomas of the colon and cervix (1 patient).


Structural analysis of unstable intermediate and stable forms of recombinant fowlpox virus.
The stability and structure of the products of recombination in a fowlpox virus (FPV) system using the thymidine kinase (TK) gene as the insertion site were examined. A 4.6 kb chimeric DNA fragment from the pUV1 expression vector, containing the bacterial lacZ gene and the vaccinia virus P7.5 promoter, was ligated into the XbaI site of the FPV TK gene. The resulting vector, pFTKlacZb, was transfected into chicken embryo fibroblast cultures infected with FPV at an m.o.i. of 0.1. Recombinants were screened for the expression of beta-galactosidase. Five recombinants were isolated and plaque-purified to 80 to 90% for expression of beta-glucosidase. Serial cell culture passage of the recombinants led to the gradual reappearance of the non-recombinant parental phenotype. Southern hybridization analysis of EcoRI fragments from all five recombinants indicated that a single cross-over homologous recombination had occurred between either the 5' or the 3' end fragments of the TK gene, generating unstable intermediate recombinants incorporating the entire pFTKlacZb vector. Secondary intermolecular or intramolecular recombination of intergenic repetitive sequences within the intermediate recombinants appears to have resulted in frequent regeneration of the parental genotype and an infrequent generation of more stable recombinants. A method was developed to select stable recombinants by passage of the intermediate recombinants in chicken embryo fibroblast cultures treated with 5-bromo-2'-deoxyuridine.


Mirrors movement-induced equivalent rotation effect in ring laser gyros.
In this letter, the relationship between the change of the closed-loop optical path and the movement of two adjacent spherical mirrors in ring laser gyros is investigated by matrix optical approach. When one spherical mirror is pushed forward and the other is pulled backward to maintain the total length of the closed-loop optical path constant, an equivalent rotation of the closed-loop optical path is found for the first time. Both numerical simulations and experimental results show the equivalent rotation rate is proportional to the velocities of the mirrors' movement.


Antifungal and Phytotoxic Activities of Isolated Compounds from Helietta parvifolia Stems.
The identification of natural and environmentally friendly pesticides is a key area of interest for the agrochemical industry, with many potentially active compounds being sourced from numerous plant species. In this study, we report the bioassay-guided isolation and identification of phytotoxic and antifungal compounds from the ethyl acetate extract of Helietta parvifolia stems. We identified eight compounds, consisting of two coumarins and six alkaloids. Among these, a new alkaloid, 2-hydroxy-3,6,7-trimethoxyquinoline-4-carbaldehyde (6), was elucidated, along with seven known compounds. The phytotoxicity of purified compounds was evaluated, and chalepin (4) was active against Agrostis stolonifera at 1 mM with 50% inhibition of seed germination and it reduced Lemna pausicotata (duckweed) growth by 50% (IC50) at 168 μM. Additionally, we evaluated the antifungal activity against the fungal plant pathogen Colletotrichum fragariae using a thin-layer chromatography bioautography assay, which revealed that three isolated furoquinoline alkaloids (flindersiamine (3), kokusagenine (7), and maculine (8)) among the isolated compounds had the strongest inhibitory effects on the growth of C. fragariae at all tested concentrations. Our results indicate that these active natural compounds, i.e., (3), (4), (7), and (8), could be scaffolds for the production of more active pesticides with better physicochemical properties.


[MOBILE TRAUMA SIMULATION IN AN EMERGENCY DEPARTMENT OF A RURAL HOSPITAL IN A CONFLICT AREA IN ISRAEL].
In recent years the city of Eilat has come under threat of rocket attacks. This emphasized the need for an upgrade of the city's medical center capabilities in trauma care. Medical simulation has been used for many years in training and for the improvement of trauma care both at the field Level and in hospitals. Although there was significant improvement in the technological aspects of simulations, the use of simulators was restricted mainly to simulation centers and its effectiveness in training and assessment of trauma teams in situ in emergency departments was still not examined. To train the personnel engaged in trauma care in a small rural medical center in the fastest and most comprehensive manner, using a mobile and highly sophisticated medical simulator. A team of simulation specialists from MSR, Israel Center for Medical Simulation, held a one week long course including all the staff members involved in trauma care, 38 doctors and nurses in total. All drills were recorded and then reviewed in the debriefing. Four staff members were trained in video debriefing at MSR. The participants were requested to complete questionnaires before and after training. Subjective feelings of competence in the team's ability to treat trauma patients were found to improve after training. Airway management rating of very good improved from 21.05% prior to training to 50% after the course. Chest trauma management and hemorrhage control scores of very good rose from 10.53% and 26.32% before to 42.11% and 55.26% respectively after the course. The highest improvement was measured in teamwork performance, which was ranked as very good: 10.53% before training and elevated to 47.37% after training. A total of 74% of the responders stated that this training program contributed very significantly to their trauma care capabilities. The use of mobile medical simulations combined with instructors from within the hospital enabled the training and assessment of the trauma care in situ and in a short time.


Ubiquitination and degradation of CFTR by the E3 ubiquitin ligase MARCH2 through its association with adaptor proteins CAL and STX6.
Golgi-localized cystic fibrosis transmembrane conductance regulator (CFTR)-associated ligand (CAL) and syntaxin 6 (STX6) regulate the abundance of mature, post-ER CFTR by forming a CAL/STX6/CFTR complex (CAL complex) that promotes CFTR degradation in lysosomes. However, the molecular mechanism underlying this degradation is unknown. Here we investigated the interaction of a Golgi-localized, membrane-associated RING-CH E3 ubiquitin ligase, MARCH2, with the CAL complex and the consequent binding, ubiquitination, and degradation of mature CFTR. We found that MARCH2 not only co-immunoprecipitated and co-localized with CAL and STX6, but its binding to CAL was also enhanced by STX6, suggesting a synergistic interaction. In vivo ubiquitination assays demonstrated the ubiquitination of CFTR by MARCH2, and overexpression of MARCH2, like that of CAL and STX6, led to a dose-dependent degradation of mature CFTR that was blocked by bafilomycin A1 treatment. A catalytically dead MARCH2 RING mutant was unable to promote CFTR degradation. In addition, MARCH2 had no effect on a CFTR mutant lacking the PDZ motif, suggesting that binding to the PDZ domain of CAL is required for MARCH2-mediated degradation of CFTR. Indeed, silencing of endogenous CAL ablated the effect of MARCH2 on CFTR. Consistent with its Golgi localization, MARCH2 had no effect on ER-localized ΔF508-CFTR. Finally, siRNA-mediated silencing of endogenous MARCH2 in the CF epithelial cell line CFBE-CFTR increased the abundance of mature CFTR. Taken together, these data suggest that the recruitment of the E3 ubiquitin ligase MARCH2 to the CAL complex and subsequent ubiquitination of CFTR are responsible for the CAL-mediated lysosomal degradation of mature CFTR.


[Glossary of bioethics terms frequently used in nutrition support].
Bioethical decisions are present in every clinical decision. Nutrition support participates the same situation. Feeding critically ill patients, etriminal patients or in permanent vegetative status is almost always involved in bioethical dilemmas. A common problem is the confusion in concepts regarding bioethics. This lack of uniformity does not help in the deliberation process. From the Working Group in Bioethics of the Spanish Society for Parenteral and Enteral Nutrition Support (SENPE) it has been considered to gather the commonest terms used in our academic area. Each term is accompanied by a definition, a description or a commentary related to its main application.


Congenital megacystis: a case report.
Congenital megacystis without other anomalies is extremely rare. A female newborn, delivered by cesarean section at the gestational age of 36 weeks, was seen. Her prenatal sonogram at 28 weeks' gestation had demonstrated a large cystic lesion in the abdomen. Abdominal distention with a palpable mass, difficult feeding and vomiting were noted after delivery. Abdominal computed tomography scan revealed a large cyst in the middle abdomen without any other anomaly. Exploratory laparotomy was performed on the fourth day of life, and showed a large bladder with normal trigon and normal ureteral insertion. There was no abnormality of other abdominal organs. Reductive cystoplasty was performed and the post-operative course was uneventful. The infant was doing well after an eight-month follow-up. Although congenital megacystis is rare, it should be considered in the differential diagnosis of any intraabdominal cystic lesion demonstrated in a prenatal sonogram. The etiology is obscure, and the benefit of reductive cystoplasty needs more cases to be confirmed.


Crossing grain boundaries in metals by slip bands, cleavage and fatigue cracks.
The size and the character (low and large angle, special boundaries, tilt and twist boundaries, twins) of the grain boundaries (GBs) in polycrystalline materials influence their strength and their fracture toughness. Recent studies devoted to nanocrystalline (NC) materials have shown a deviation from the Hall-Petch law. Special GBs formed by Σ3 twins in face-centred cubic metals are also known to have a strong effect on the mechanical behaviour of these metals, in particular their work-hardening rate. Grain orientation influences also crack path, the fracture toughness of body-centred cubic (BCC) metals and the fatigue crack growth rate of microstructurally short cracks. This paper deals both with slip transfer at GBs and with the interactions between propagating cracks with GBs. In the analysis of slip transfer, the emphasis is placed on twin boundaries (TBs) for which the dislocation reactions during slip transfer are analysed theoretically, experimentally and using the results of atomic molecular simulations published in the literature. It is shown that in a number of situations this transfer leads to a normal motion of the TB owing to the displacement of partial dislocations along the TB. This motion can generate a de-twinning effect observed in particular in NC metals. Crack propagation across GBs is also considered. It is shown that cleavage crack path behaviour in BCC metals is largely dependent on the twist component of the GBs. A mechanism for the propagation of these twisted cracks involving a segmentation of the crack front and the existence of intergranular parts is discussed and verified for a pressure vessel steel. A similar segmentation seems to occur for short fatigue cracks although, quite surprisingly, this crossing mechanism for fatigue cracks does not seem to have been examined in very much detail in the literature. Metallurgical methods used to improve the strength of the materials, via grain boundaries, are briefly discussed.


Schools' Cafeteria Status: Does it Affect Snack Patterns? A Qualitative Study.
The aim of this study was to evaluate patterns of students' snacks habits regarding to their schools' cafeteria status in Tehran by focus group discussion (FGD) technique. Participants were 240 students (12-15 years old), selected from12 middle-schools in Tehran. The field study consisted of 24 FGDs sessions; involving 8-10 participants. Collected data were coded, categorized and analyzed using constant comparative method. Over half of the students believed that snack consumption is necessary. Although, majority of students believed that their schools' cafeterias are not acceptable, they noted them as one of the necessary parts of school. Nearly half of the children were complaining of unvaried and expensive food items. The most purchased items were: Cookies, sandwiches with mayonnaise and ketchup, soft drinks and chocolate milk. Most of the students were interested in having roles in their cafeterias. Schools' cafeteria are significant sources of supplying adolescents' snacks, so developing hygienic stores containing healthy and nutritious food items is a key element to affect their snack selection positively. Reaching this goal requires a multi disciplinary approach through participation of students, school staff, parents, and the support of community and media.


Fracture prediction in rheumatoid arthritis: validation of FRAX with bone mineral density for incident major osteoporotic fractures.
FRAX® uses clinical risk factors, with or without bone mineral density (BMD), to calculate 10-year fracture risk. Rheumatoid arthritis (RA) is a risk factor for osteoporotic fracture and a FRAX input variable. FRAX predates the current era of RA treatment. We examined how well FRAX predicts fracture in contemporary RA patients. Administrative data from patients receiving BMD testing were linked to the Manitoba Population Health Research Data Repository. Observed cumulative 10-year Major Osteoporotic Fracture (MOF) probability was compared with FRAX-predicted 10-year MOF probability with BMD for assessing calibration. MOF risk stratification was assessed using Cox regression. RA patients (N = 2,099, 208 with incident MOF) and non-RA patients (N = 2,099, with 165 incident MOF) were identified. For RA patients, FRAX predicted 10-year risk was 13.2% and observed 10-year MOF risk was 13.2% (95% CI 11.6% to 15.1%). The slope of the calibration plot was 0.67 (95% CI 0.53-0. 81) in those with RA vs 0.98 (95% CI 0.61-1.34) in non-RA patients. Risk was overestimated in RA patients with high FRAX scores (>20%), but FRAX was well-calibrated in other groups. FRAX stratified risk in those with and without RA (hazard ratios 1.52, 95% 1.25-1.72 vs 2.00, 95% 1.73-2.31), with slightly better performance in the latter (p-interaction = 0.004). FRAX predicts fracture risk in contemporary RA patients but may slightly overestimate risk in those already at high predicted risk. Thus, the current FRAX tool continues to be appropriate for fracture risk assessment in RA patients.


Investigating Changes in Real-time Conscious Postural Processing by Older Adults during Different Stance Positions Using Electroencephalography Coherence.
Background/Study Context: Adjustments of posture in response to balance challenges may lead to subsequent increases in conscious posture processing. If cognitive resources are stretched by conscious processing of postural responses fewer resources will be available to attend to environmental trip or fall hazards. The objective of the study was to explore brain activity related to conscious processing of posture as a function of movement specific reinvestment and fear of falling. Method: Forty-three older adults (M = 71.4, SD = 4.1) stood with a wide or narrow stance on a force-plate while neural coherence between verbal-analytical (T3) and motor planning (Fz) regions of the brain was assessed using electroencephalography. The propensity for movement specific reinvestment was assessed using the Chinese version Movement Specific Reinvestment Scale (MSRS-C) and fear of falling was assessed using the Chinese version Fall Efficacy Scale International (FES-I[CH]). Results: Scores from the MSRS-C were negatively correlated with changes in T3-Fz coherence that occurred when participants shifted from wide to narrow stance. Together, MSRS-C and FES-I(CH) uniquely predicted the percentage change in T3-Fz coherence between the two stance conditions. Conclusion: Presented with two postural tasks of different complexities, participants with a lower propensity for conscious control of their movements (movement specific reinvestment) exhibited larger changes in real-time brain activity (neural coherence) associated with conscious postural processing.


Clinical and MRI Features of Posterior Reversible Encephalopathy Syndrome With Atypical Regions: A Descriptive Study With a Large Sample Size.
Background: Accurate diagnosis and timely treatment for posterior reversible encephalopathy syndrome (PRES) with atypical regions are very important in clinical practice. However, until now, little has been known about the clinical and MRI manifestations of this disease. Therefore, the aim of this study is to investigate the clinical and MRI features of PRES to promote clinical management and deepen our understanding of this disease. Materials and Methods: Data from six PRES patients with atypical regions were collected from our hospital. Data from another 550 cases were obtained by searching the PubMed, EMBASE and Web of Science databases with the keywords "posterior reversible encephalopathy syndrome" "PRES" "reversible posterior leukoencephalopathy" "RPLS" "hypertensive encephalopathy" "hyperperfusion encephalopathy" or "reversible posterior cerebral edema encephalopathy." The clinical and MRI features of these 556 cases were analyzed together. Results: A total of 305 patients were female, and 248 were male, with a median age of 34 years. The information on sex and age of three patients was not available. The most common symptom was headache (282/556, 50.7%), followed by altered mental status (243/556, 43.7%), seizures (233/556, 41.9%), visual disturbances (194/556, 34.9%), nausea/vomiting (130/556, 23.4%), and focal neurological deficits (101/556, 18.2%). Hypertension (425/556, 76.4%), renal diseases (152/556, 27.3%), immunosuppressant drugs (79/556, 14.2%), and chemotherapy/chemoradiotherapy (59/556, 10.6%) were the major predisposing factors. The atypical regions of the lesions were the cerebellum (331/556, 59.5%), basal ganglia (135/556, 24.3%), periventricular/deep white matter (125/556, 22.5%), pons (124/556, 22.3%), brainstem (115/556, 20.7%), thalamus (114/556, 20.5%), midbrain (48/556, 8.6%), spinal cord (33/556, 5.9%), and medulla (29/556, 5.2%). Additionally, the following typical regions were observed: occipital (278/556, 50.0%), parietal (234/556, 42.1%), frontal (150/556, 27.0%), and temporal (124/556, 22.3%) lobes. The major treatments were antihypertensives (358/515, 69.5%), antiepileptics/sedation (126/515, 24.5%), discontinuation/switching agents (67/515, 13.0%), and steroids (54/515, 10.5%). The median time of the clinical state improved and abnormal neuroimaging resolved is 2-3 weeks after appropriate treatment. Conclusion: The common symptoms of PRES with atypical regions include headaches, altered mental status, seizures, visual disturbances, nausea or vomiting, and focal neurological deficits. The frequent predisposing factors include hypertension, renal diseases, immunosuppressant drugs and chemotherapy/chemoradiotherapy. MRI features are mainly characterized by vasogenic edema in central zones always accompanied by typical regions. Most cases can be reversed in 2-3 weeks when promptly recognized and properly treated.


CMR Reclassifies the Majority of Patients with Suspected MINOCA and non MINOCA.
In approximately 5-15% of all cases of acute coronary syndromes (ACS) have unobstructed coronaries on angiography. Cardiac magnetic resonance (CMR) has proven useful to identify in most patients the underlying diagnosis associated with this presentation. However, the role of CMR to reclassify patients from the initial suspected condition has not been clarified. The aim of this study was to assess the proportion of patients with suspected MINOCA, or non-MINOCA, that CMR reclassifies with an alternative diagnosis from the original clinical suspicion. A retrospective cohort of patients in a tertiary cardiology centre was identified from a registry database. Patients who were referred for CMR for investigation of suspected MINOCA, and a diagnosis pre- and post-CMR was recorded to determine the proportion of diagnoses reclassified.A total of 888 patients were identified in the registry. CMR reclassified diagnosis in 78% of patients. Diagnosis of MINOCA was confirmed in only 243 patients (27%), whilst most patients had an alternative diagnosis (73%): myocarditis n = 217 (24%), Takotsubo syndrome n = 115 (13%), cardiomyopathies n = 97 (11%), normal CMR/non-specific n = 216 (24%). In a large single-centre cohort of patients presenting with acute coronary syndrome and unobstructed coronary arteries, most patients had a non-MINOCA diagnosis (73%) (myocarditis, TakoTsubo, cardiomyopathies or normal CMR/non-specific findings), whilst only a minority had confirmed MINOCA (27%). Performing CMR led to reclassifying patients' diagnosis in 78% of cases, thus confirming its important clinical role and underscoring the clinical challenge in diagnosing MINOCA and non MINOCA conditions.


A questionnaire-based survey of errors in diagnostic histopathology throughout the United Kingdom.
To obtain a crude estimate of the rate at which consultant histopathologists become aware of errors in their work, and to gather information about the circumstances under which errors are made. A postal questionnaire was sent to all consultant histopathologists (1021) in the United Kingdom. There was a 19.8% response to the questionnaire (202 returned). 119 pathologists reported 244 errors, 82 from within the preceding year. 42% of these errors had affected patient management. Pathologists usually blamed human error or excessive workload; however, data analysis did not reveal an excess of responses from laboratories with heavy workloads. A disproportionately large proportion of errors appear to be made in samples from the lymphoreticular system and by locum consultant pathologists. This study undoubtedly suffered from reporting bias, but the figures suggest that a typical pathologist probably becomes aware of having made a serious diagnostic error approximately once per year. A larger, more systematic study of the circumstances under which errors are likely to occur appears to be justified but would require significant resources because of the large sample size required.


Bisphosphonates inhibit IL-6 production by human osteoblast-like cells.
Since bisphosphonates prevent bone loss in osteoporosis and rheumatoid arthritis, diseases in which the osteoclastogenic and inflammatory cytokine interleukin-6 plays a pathophysiologic role, we studied whether these drugs regulate the production of this cytokine by osteoblasts. Spontaneous and IL-1 + TNF-alpha stimulated IL-6 release was measured in supernatants of cultures of human osteoblastic osteosarcoma cells MG-63, pretreated for 4 hours with different doses of etidronate, clodronate or alendronate using a specific bioassay. Etidronate [from 10(-4) to 10(-8) M] or alendronate [from 10(-6) to 10(-11) M] inhibited in a dose-dependent manner the cytokine-induced IL-6 secretion [60+/-9.5% at 10(-5) M and 65+/-12% at 10(-7) M, respectively; p < 0.01]. Though significant, the inhibitory effect of clodronate was less [35+/-7% at 10(-5) M, p < 0.05]. These in vitro observations might have in vivo relevance in explaining at least in part the mechanisms by which bisphosphonates inhibit systemic and periarticular bone resorption.


Ultrasound-guided regional anesthesia performance in the early learning period: effect of simulation training.
Success in performing ultrasound-guided peripheral nerve blockade (PNB) demands sound knowledge of sonoanatomy, good scanning techniques, and proper hand-eye coordination. The objectives of our study were to evaluate whether simulator training aids success of novice operators in ultrasound-guided PNB and to determine what number of procedures is required to attain proficiency. Twenty Postgraduate Year 2 anesthesiology residents with no previous experience in ultrasound-guided PNB were randomly assigned into 2 groups. Both groups received conventional teaching comprising of 4 didactic lectures on PNBs with ultrasound guidance. Using a low-fidelity simulation model, 1 group further received an hourlong training session on needling and proper hand-eye coordination. Once the training was over, the residents started their rotation through our block room. Using a logbook, each resident recorded the number of successful and failed ultrasound-guided regional anesthesia blocks performed over a 3-week period. A successful block was defined as one that was effective for surgical anesthesia and performed within 15 mins, with only verbal guidance from a staff anesthesiologist. Cumulative summation charts were created to track progress using a predetermined acceptable failure rate of 30%. The conventional training group had 98 successful blocks, and the simulation group had 144 (51.3% vs 64%; P = 0.016). In the conventional training group, 4 of 10 residents achieved proficiency, and in the simulation training group, 8 of 10 residents achieved proficiency (80% vs 40%; P = 0.0849). Simulation training improves success rate in ultrasound-guided performance of regional anesthesia.


Boring in response to bark and phloem extracts from North American trees does not explain host acceptance behavior of Orthotomicus erosus (Coleoptera: Scolytidae).
When invasive herbivorous insects encounter novel plant species, they must determine whether the novel plants are hosts. The Mediterranean pine engraver, Orthotomicus erosus (Wollaston), an exotic bark beetle poised to expand its range in North America, accepts hosts after contacting the bark. To test the hypothesis that O. erosus accepts hosts on the basis of gustatory cues, we prepared bark and phloem extracts from logs of four North American tree species that we had used in previous host acceptance experiments. Water, methanol, and hexane extracts of red pine, tamarack, balsam fir, and paper birch were presented alone and in combination on a neutral filter paper substrate in a section of a plastic drinking straw. Boring behavior in response to the three-extract combinations differed from the pattern of acceptance previously observed among species when the beetles were in contact with the bark surface. Only the aqueous extracts of tamarack, Larix laricina, increased the initiation and the extent of boring by O. erosus on the filter paper substrate. We conclude that the effects of extracted chemicals do not match the behavior of the beetles observed when penetrating excised bark and phloem discs, indicating that host selection by O. erosus may not be predictable from bark and phloem chemistry alone. Instead, host acceptance may be determined by nongustatory stimuli or by a combination of stimuli including gustatory and nongustatory cues.


Predictive validity and measurement issues in documenting quit intentions in population surveillance studies.
Discrete classification of smokers by intention to quit is desirable in many public health and clinical settings. Two methodological studies examine measurement properties of measures of discrete-time intention to quit smoking used in population-based tobacco surveillance surveys: an ecological comparison of rates of positive intention in relation to the form of measure used and a prospective analysis examining predictive validity of self-reported quit intentions using multiple possible points of dichotomization of an ordinal measure of intention to quit. The prospective analysis used a repeated measures design and follow-up to 1 year for 2,047 smokers in the Ontario Tobacco Survey cohort. The estimated percent of smokers intending to quit was significantly higher using the Stages of Change intention measure, relative to another single question measure. Significant dose-response effects were found. The sooner one intended to quit the more likely one was to make an attempt or achieve at least 30 days abstinence in the next 6 months. Intending to quit in a month or later was not associated with cessation during follow-up among respondents without prior attempts. Examination of cutpoints revealed no value, which maximized both positive and negative prediction. Regardless of quit attempt history, greatest predictive validity was found where respondents stated that they had no intention at all. Measures of intentions quit smoking in specific time periods and expressed as dichotomies have limited psychometric properties but utility in applied research. Our findings suggest a possible measurement effect warranting caution in comparisons across studies.


The regulation of respiratory resistance in exercising horses.
Horses display remarkable aerobic capabilities, attaining during muscular exercise a maximal rate of oxygen consumption about 30-fold higher than the resting value, and 2.5-fold higher than that of other mammals of similar body mass. Under these circumstances an enormous mechanical burden is expected to impinge on the equine respiratory pump and regulatory mechanisms aiming to minimize this load may play an important role in determining the adequacy of the respiratory system to the metabolic requirements. The behaviour of the respiratory system has been investigated in horses at rest and during treadmill locomotion at different velocities and gaits. During exercise hyperpnoea, horses exhibit a significant reduction in the lung viscous resistance not observed in other mammals, such as dogs and humans. Therefore, the exercise-dependent increase in the rate of mechanical work of breathing is lower in the horse than in other mammals. This increase in the equine airway patency during exercise appeared to be mainly determined by the pattern of laryngeal movements. In fact, during exercise, the laryngeal cross-sectional area, determined with a video-endoscopic imaging technique at the level of rima glottidis (CSArg), undergoes during inspiration an increase averaging up to over 4 times the resting expiratory values. Although a significant linear correlation was found between CSArg and minute ventilation (VE), the laryngeal activation contributes to increase lung conductance only when CSArg is narrower than the tracheal section. It appears therefore that in exercising horses pulmonary resistive features are finely controlled to reduce the mechanical load supported by the respiratory muscles and to counterbalance the increase in the ventilatory energetic requirements inherent in the remarkably enhanced aerobic performance observed in this species.


Molecular detection and characterization of Hepatozoon canis in stray dogs from Cuba.
Canine hepatozoonosis caused by Hepatozoon canis is a worldwide distributed tick-borne disease of domestic and wild canids that is transmitted by ingestion of Rhipicephalus sanguineus sensu lato (s.l.) ticks. The present study was aimed to determine the prevalence of Hepatozoon infections in 80 stray dogs from Havana Province in Cuba, and to confirm the species identity and phylogenetic relationships of the causative agent. Samples were screened by microscopical examination of thin blood smears for the presence of Hepatozoon spp. gamonts and by genus-specific SYBR green-based real-time PCR assay targeting the 18S rRNA gene. Direct microscopy examination revealed Hepatozoon gamonts in the peripheral blood of 8 dogs (10.0%; 95% CI: 4.80-18.0%), while 38 animals (47.5%; 95% CI: 36.8-58.4%) were PCR-positive, including all microscopically positive dogs. Hence, the agreement between the two detection methods was 'poor' (κ = 0.20). Hematological parameters did not differ significantly between PCR-positive and PCR-negative dogs (p > 0.05). The DNA sequences of the 18S rRNA gene of the Hepatozoon spp. from Cuban dogs showed a nucleotide identity >99% with those of 18S rRNA sequences of Hepatozoon canis isolates from Czech Republic, Brazil and Spain. Phylogenetic analysis revealed that obtained sequences clustered within the Hepatozoon canis clade, different from the Hepatozoon felis or Hepatozoon americanum clades. The present study represents the first molecular characterization of Hepatozoon canis in stray dogs within Cuba.


Novel Antimicrobial Peptides EeCentrocins 1, 2 and EeStrongylocin 2 from the Edible Sea Urchin Echinus esculentus Have 6-Br-Trp Post-Translational Modifications.
The global problem of microbial resistance to antibiotics has resulted in an urgent need to develop new antimicrobial agents. Natural antimicrobial peptides are considered promising candidates for drug development. Echinoderms, which rely on innate immunity factors in the defence against harmful microorganisms, are sources of novel antimicrobial peptides. This study aimed to isolate and characterise antimicrobial peptides from the Edible sea urchin Echinus esculentus. Using bioassay-guided purification and cDNA cloning, three antimicrobial peptides were characterised from the haemocytes of the sea urchin; two heterodimeric peptides and a cysteine-rich peptide. The peptides were named EeCentrocin 1 and 2 and EeStrongylocin 2, respectively, due to their apparent homology to the published centrocins and strongylocins isolated from the green sea urchin Strongylocentrotus droebachiensis. The two centrocin-like peptides EeCentrocin 1 and 2 are intramolecularly connected via a disulphide bond to form a heterodimeric structure, containing a cationic heavy chain of 30 and 32 amino acids and a light chain of 13 amino acids. Additionally, the light chain of EeCentrocin 2 seems to be N-terminally blocked by a pyroglutamic acid residue. The heavy chains of EeCentrocins 1 and 2 were synthesised and shown to be responsible for the antimicrobial activity of the natural peptides. EeStrongylocin 2 contains 6 cysteines engaged in 3 disulphide bonds. A fourth peptide (Ee4635) was also discovered but not fully characterised. Using mass spectrometric and NMR analyses, EeCentrocins 1 and 2, EeStrongylocin 2 and Ee4635 were all shown to contain post-translationally brominated Trp residues in the 6 position of the indole ring.


The history of seabird colonies and the North Water ecosystem: Contributions from palaeoecological and archaeological evidence.
The North Water (NOW) polynya is one of the most productive marine areas of the Arctic and an important breeding area for millions of seabirds. There is, however, little information on the dynamics of the polynya or the bird populations over the long term. Here, we used sediment archives from a lake and peat deposits along the Greenland coast of the NOW polynya to track long-term patterns in the dynamics of the seabird populations. Radiocarbon dates show that the thick-billed murre (Uria lomvia) and the common eider (Somateria mollissima) have been present for at least 5500 cal. years. The first recorded arrival of the little auk (Alle alle) was around 4400 cal. years BP at Annikitsoq, with arrival at Qeqertaq (Salve Ø) colony dated to 3600 cal. years BP. Concentrations of cadmium and phosphorus (both abundant in little auk guano) in the lake and peat cores suggest that there was a period of large variation in bird numbers between 2500 and 1500 cal. years BP. The little auk arrival times show a strong accord with past periods of colder climate and with some aspects of human settlement in the area.


Sodium glucose co-transporter 2 inhibitor luseogliflozin in the management of type 2 diabetes: a drug safety evaluation.
Sodium glucose co-transporter-2 (SGLT2) inhibitors have been developed recently as a new class of anti-diabetic drug, and are becoming widely used in the management of type 2 diabetes (T2D). As these agents have a considerably different glucose-lowering mechanism from those of other anti-diabetic drugs, safe use of this drug class needs to be discussed based on data available from preapproval clinical trials as well as real-world studies. The SGLT2 inhibitor luseogliflozin was developed by Taisho Pharmaceutical Co., Ltd. and was approved as an oral anti-diabetic drug for T2D in Japan Areas covered: The overall safety and efficacy of SGLT2 inhibitor luseogliflozin are summarized on the basis of a literature review, with a focus on reported adverse drug reactions in preapproval clinical trials and a post-marketing surveillance. Expert opinion: SGLT2 inhibitor luseogliflozin is well tolerated, significantly improves hyperglycemia in preapproval clinical trials, and has a favorable safety profile in both preapproval clinical trials and post-marketing surveillance in elderly patients. While long-term safety and efficacy remain to be seen, luseogliflozin can benefit T2D patients worldwide. However, healthcare professionals must perform appropriate patient education that includes temporary withdrawal of luseogliflozin during patient a 'sick day' and avoidance of strict carbohydrate restriction during luseogliflozin treatment.


The fate of tetrathionate during the development of a biofilm in biogenic sulfuric acid attack on different cementitious materials.
The biodeterioration of cement-based materials in sewer environments occurs because of the production of sulfuric acid from the biochemical oxidation of H2S by sulfur-oxidizing bacteria (SOB). In the perspective of determining the possible reaction pathways for the sulfur cycle in such conditions, hydrated cementitious binders were exposed to an accelerated laboratory test (BAC test) to reproduce a biochemical attack similar to the one occurring in the sewer networks. Tetrathionate was used as a reduced sulfur source to naturally develop sulfur-oxidizing activities on the surfaces of materials. The transformation of tetrathionate was investigated on materials made from different binders: Portland cement, calcium aluminate cement, calcium sulfoaluminate cement and alkali-activated slag. The pH and the concentration of the different sulfur species were monitored in the leached solutions during 3 months of exposure. The results showed that the formation of different polythionates was independent of the nature of the material. The main parameter controlling the phenomena was the evolution of the pH of the leached solutions. Moreover, tetrathionate disproportionation was detected with the formation of more reduced forms of sulfur compounds (pentathionate, hexathionate and elemental sulfur) along with thiosulfate and sulfate. The experimental findings allowed numerical models to be developed to estimate the amount of sulfur compounds as a function of the pH evolution. In addition, biomass samples were collected from the exposed surface and from the deteriorated layers to identify the microbial populations. No clear influence of the cementitious materials on the selected populations was detected, confirming the previous results concerning the impact of the materials on the selected reaction pathways for tetrathionate transformation.


Intervertebral disc tissue engineering using a novel hyaluronic acid-nanofibrous scaffold (HANFS) amalgam.
Degeneration of the intervertebral disc (IVD) represents a significant musculoskeletal disease burden. Although spinal fusion has some efficacy in pain management, spine biomechanics is ultimately compromised. In addition, there is inherent limitation of hardware-based IVD replacement prostheses, which underscores the importance of biological approaches to disc repair. In this study, we have seeded multipotent, adult human mesenchymal stem cells (MSCs) into a novel biomaterial amalgam to develop a biphasic construct that consisted of electrospun, biodegradable nanofibrous scaffold (NFS) enveloping a hyaluronic acid (HA) hydrogel center. The seeded MSCs were induced to undergo chondrogenesis in vitro in the presence of transforming growth factor-beta for up to 28 days. The cartilaginous hyaluronic acid-nanofibrous scaffold (HANFS) construct architecturally resembled a native IVD, with an outer annulus fibrosus-like region and inner nucleus pulposus-like region. Histological and biochemical analyses, immunohistochemistry, and gene expression profiling revealed the time-dependent development of chondrocytic phenotype of the seeded cells. The cells also maintain the microarchitecture of a native IVD. Taken together, these findings suggest the prototypic potential of MSC-seeded HANFS constructs for the tissue engineering of biological replacements of degenerated IVD.