13C-NMR and transient kinetic studies on lactate dehydrogenase [Cys(13CN)165]. Direct measurement of a rate-limiting rearrangement in protein structure.
Chemical modification of cysteine-165 in pig heart lactate dehydrogenase to produce lactate dehydrogenase [Cys(13CN)165] introduces an covalently bound, enriched 13C probe at a position adjacent to the active cen. The signal from the thiocyanate probe is clearly visible at 47 ppm relative to dioxane. On formation of binary complexes with NAD+ and NADH, no signal change is detected. Formation of the ternary complexes E-NADH-oxamate and E-NAD+-oxalate results in an upfield shift of the signal of 1.2 ppm. These results interpreted as demonstrating that binding of the substrate analogue induces a conformational change a position adjacent to the active centre. Exchange experiments in which the enzyme is poised in dynamic equilibrium between binary and ternary complexes show that the rate at which the probe senses a change environment is the same as the kinetically observed unimolecular event which limits the enzyme-catalyst reduction of pyruvate. The two processes show the same dependence on temperature, solvent composition and pH. These results indicate that the rate-limiting isomerisation corresponds to a rearrangement of the protein in the region of cysteine-165.


Instrument validation and measurement of home healthcare nurses' knowledge of artificial nutrition and hydration at end of life.
The purpose of this exploratory pilot study was two-fold: (a) to determine the reliability of the instrument to measure nurses' knowledge of artificial nutrition and hydration, and (b) to assess home healthcare nurses' knowledge of artificial nutrition and hydration. A cross-sectional design was used to administer the questionnaire to 91 home healthcare nurses in North Carolina; 33 questionnaires were returned. Results revealed an accurate answer rate of 73% and Cronbach's alpha was 0.71, indicating adequate internal consistency reliability.


The occurrence of salmonellae in bean sprouts in Thailand.
Detection of Salmonella spp. in 344 samples of bean sprouts was attempted by pre-enriched in buffered peptone water (BPW) or lactose broth (LB) at 37 degrees C, followed by selective enrichments, selenite cystein broth (SCB) and Rappaport medium (RV) at 43 degrees C. Four selective media, bismuth sulphite agar (BSA), brilliant green agar (BGA), Salmonella-Shigella agar (SSA) and MacConkey agar (MA) were used for isolation. Salmonellae were isolated from 30 (8.7%) samples. They were serotypes lexington (56.7%), orion (16.7%), senftenberg (16.7%), tennessee (3.3%), poona (3.3%) and weltevreden (3.3%). Pre-enrichment in LB, followed by enrichment in SCB and streaking on to BSA gave the highest recoveries. Fifteen of 30 strains isolated were sensitive to all 14 antibiotics tested. Seven strains were resistant to one, and a strains showed multiple resistance to 2-4 antimicrobial agents.


Blast injury research models.
Blast injuries are an increasing problem in both military and civilian practice. Primary blast injury to the lungs (blast lung) is found in a clinically significant proportion of casualties from explosions even in an open environment, and in a high proportion of severely injured casualties following explosions in confined spaces. Blast casualties also commonly suffer secondary and tertiary blast injuries resulting in significant blood loss. The presence of hypoxaemia owing to blast lung complicates the process of fluid resuscitation. Consequently, prolonged hypotensive resuscitation was found to be incompatible with survival after combined blast lung and haemorrhage. This article describes studies addressing new forward resuscitation strategies involving a hybrid blood pressure profile (initially hypotensive followed later by normotensive resuscitation) and the use of supplemental oxygen to increase survival and reduce physiological deterioration during prolonged resuscitation. Surprisingly, hypertonic saline dextran was found to be inferior to normal saline after combined blast injury and haemorrhage. New strategies have therefore been developed to address the needs of blast-injured casualties and are likely to be particularly useful under circumstances of enforced delayed evacuation to surgical care.


Mesalazine to treat symptomatic uncomplicated diverticular disease and to prevent acute diverticulitis occurrence. A systematic review with meta-analysis of randomized, placebo-controlled trials.
Symptomatic Uncomplicated Diverticular disease (SUDD) affects about 25% of patients harboring colonic diverticula. We assessed the effectiveness of mesalazine in improving symptoms (namely abdominal pain, primary outcome) and in preventing diverticulitis occurrence (secondary outcome) in patients with SUDD. Pertinent studies were selected from the Medline and the Cochrane Central Register of Controlled Trials. Only randomized clinical trials (RCTs) (irrespective of language, blinding, or publication status), which compared mesalazine, irrespective of the dosage assumption, with placebo in SUDD were evaluated. Four RCTs enrolled 379 patients, 197 treated with mesalazine and 182 with placebo. Two studies provided data on symptom relief according to definition: it was achieved in 97/121 (80%) patients in the mesalazine group and in 81/129 (62.7%) patients in the placebo group (OR 0.43; 95% CI 0.24-0.75; p=0.003 in favour of the mesalazine group). Two studies provided information regarding occurrence of diverticulitis during follow-up. It occurred in 23/119 (19.3%) patients in the mesalazine group and in 34/102 (33.3%) patients in the placebo group (OR 0.35; 95% CI 0.17-0.70; p=0.003 in favour of the mesalazine group). Treatment with mesalazine seems to be effective in achieving symptom relief and in the primary prevention of diverticulitis in patients with SUDD.


Getting sperm and egg together: things conserved and things diverged.
Sperm-egg interactions occur at multiple levels on the egg surface, first with the egg's extracellular matrix and then with the egg's plasma membrane. The BioPore minisymposium on "The Egg Surface" at the 1999 annual meeting of the Society for the Study of Reproduction highlighted a series of events underlying successful interactions of the sperm with the egg: 1) composition, synthesis, and assembly of the mouse egg's extracellular matrix, the zona pellucida, during oogenesis; 2) oocyte maturation and development of the sperm-binding domain of mouse eggs; and 3) characterization of functional domains in different sperm ligands (fertilin-alpha and fertilin-beta in the mouse and lysin in the abalone) that recognize cognate binding sites on the egg surface. Data that were presented are reviewed here and discussed with respect to conserved and divergent features of gamete functions.


[Experience of continuous educational process in training of doctors specializing in reflexotherapy].
Highlights the experience of the Department of Neurology and Reflexology to organize continuous education process in preparing doctors with specialization reflexology.


Genetic variants in EPAS1 contribute to adaptation to high-altitude hypoxia in Sherpas.
Sherpas comprise a population of Tibetan ancestry in the Himalayan region that is renowned for its mountaineering prowess. The very small amount of available genetic information for Sherpas is insufficient to explain their physiological ability to adapt to high-altitude hypoxia. Recent genetic evidence has indicated that natural selection on the endothelial PAS domain protein 1 (EPAS1) gene was occurred in the Tibetan population during their occupation in the Tibetan Plateau for millennia. Tibetan-specific variations in EPAS1 may regulate the physiological responses to high-altitude hypoxia via a hypoxia-inducible transcription factor pathway. We examined three significant tag single-nucleotide polymorphisms (SNPs, rs13419896, rs4953354, and rs4953388) in the EPAS1 gene in Sherpas, and compared these variants with Tibetan highlanders on the Tibetan Plateau as well as with non-Sherpa lowlanders. We found that Sherpas and Tibetans on the Tibetan Plateau exhibit similar patterns in three EPAS1 significant tag SNPs, but these patterns are the reverse of those in non-Sherpa lowlanders. The three SNPs were in strong linkage in Sherpas, but in weak linkage in non-Sherpas. Importantly, the haplotype structured by the Sherpa-dominant alleles was present in Sherpas but rarely present in non-Sherpas. Surprisingly, the average level of serum erythropoietin in Sherpas at 3440 m was equal to that in non-Sherpas at 1300 m, indicating a resistant response of erythropoietin to high-altitude hypoxia in Sherpas. These observations strongly suggest that EPAS1 is under selection for adaptation to the high-altitude life of Tibetan populations, including Sherpas. Understanding of the mechanism of hypoxia tolerance in Tibetans is expected to provide lights to the therapeutic solutions of some hypoxia-related human diseases, such as cardiovascular disease and cancer.


Benign breast lesions with malignant clinical and mammographic presentations.
Nine cases of benign breast disease in which mammograms had been false-positive were collected at Northwestern Memorial Hospital. In all but one case the patients had presented initially with questionable masses that required biopsies with requests for frozen section diagnoses. Included in the study were three cases of indurative mastopathy, three cases of fibrocystic disease with sclerosing adenosis, and one case each of sclerosing papillary proliferation, infarcted intraductal papilloma, and fat necrosis with foreign body giant cell reaction. The mammographic and histologic findings for all cases were reviewed. Indurative mastopathy is a poorly known entity with radiologic features highly suggestive of malignancy. As described previously (Cancer 47:561, 1981), the lesion consists of a central nidus of elastosis with irregular projections radiating into the adjacent breast tissue. Peripheral areas of the infarcted papilloma and sclerosing papillary proliferation could be confused with infiltrating carcinoma in frozen sections. Familiarity of pathologists with these lesions is essential for avoiding the overdiagnosis of carcinoma.


Pulsatility of insulin release--a clinically important phenomenon.
The mechanisms and clinical importance of pulsatile insulin release are presented against the background of more than half a century of companionship with the islets of Langerhans. The insulin-secreting beta-cells are oscillators with intrinsic variations of cytoplasmic ATP and Ca(2+). Within the islets the beta-cells are mutually entrained into a common rhythm by gap junctions and diffusible factors (ATP). Synchronization of the different islets in the pancreas is supposed to be due to adjustment of the oscillations to the same phase by neural output of acetylcholine and ATP. Studies of hormone secretion from the perfused pancreas of rats and mice revealed that glucose induces pulses of glucagon anti-synchronous with pulses of insulin and somatostatin. The anti-synchrony may result from a paracrine action of somatostatin on the glucagon-producing alpha-cells. Purinoceptors have a key function for pulsatile release of islet hormones. It was possible to remove the glucagon and somatostatin pulses with maintenance of those of insulin with an inhibitor of the P2Y(1) receptors. Knock-out of the adenosine A(1) receptor prolonged the pulses of glucagon and somatostatin without affecting the duration of the insulin pulses. Studies of isolated human islets indicate similar relations between pulses of insulin, glucagon, and somatostatin as found during perfusion of the rodent pancreas. The observation of reversed cycles of insulin and glucagon adds to the understanding how the islets regulate hepatic glucose production. Current protocols for pulsatile intravenous infusion therapy (PIVIT) should be modified to mimic the anti-synchrony between insulin and glucagon normally seen in the portal blood.


A Framework for Optimizing Phytosanitary Thresholds in Seed Systems.
Seedborne pathogens and pests limit production in many agricultural systems. Quarantine programs help prevent the introduction of exotic pathogens into a country, but few regulations directly apply to reducing the reintroduction and spread of endemic pathogens. Use of phytosanitary thresholds helps limit the movement of pathogen inoculum through seed, but the costs associated with rejected seed lots can be prohibitive for voluntary implementation of phytosanitary thresholds. In this paper, we outline a framework to optimize thresholds for seedborne pathogens, balancing the cost of rejected seed lots and benefit of reduced inoculum levels. The method requires relatively small amounts of data, and the accuracy and robustness of the analysis improves over time as data accumulate from seed testing. We demonstrate the method first and illustrate it with a case study of seedborne oospores of Peronospora effusa, the causal agent of spinach downy mildew. A seed lot threshold of 0.23 oospores per seed could reduce the overall number of oospores entering the production system by 90% while removing 8% of seed lots destined for distribution. Alternative mitigation strategies may result in lower economic losses to seed producers, but have uncertain efficacy. We discuss future challenges and prospects for implementing this approach.


Distinct oligomeric forms of the Pseudomonas aeruginosa RetS sensor domain modulate accessibility to the ligand binding site.
Bacterial two-component regulatory systems (TCSs) sense environmental stimuli to adapt the lifestyle of microbial populations. For many TCSs the stimulus is a ligand of unknown chemical nature. Pseudomonas aeruginosa utilizes the closely related RetS and LadS sensor kinases to switch between acute and chronic infections. These sensor proteins antagonistically mediate biofilm formation through communication with a central TCS, GacA/GacS. Recently, it was shown that RetS modulates the GacS sensor activity by forming RetS/GacS heterodimers. LadS and RetS are hybrid sensors with a signalling domain consisting of a 7-transmembrane (7TMR) region and a periplasmic sensor domain (diverse intracellular signalling module extracellular 2, DISMED2). The 2.65 A resolution crystal structure of RetS DISMED2, called RetSp, reveals three distinct oligomeric states capable of domain swapping. The RetSp structure also displays two putative ligand binding sites. One is equivalent to the analogous site in the structurally-related carbohydrate binding module (CBM) but the second site is located at a dimer interface. These observations highlight the modular architecture and assembly of the RetSp fold and give clues on how homodimerization of RetS could be modulated upon ligand binding to control formation of a RetS/GacS heterodimer. Modelling the DISMED2 of LadS reveals conservation of only one ligand binding site, suggesting a distinct mechanism underlying the activity of this sensor kinase.


Life goes on: the resilience of maternal primary care during the Ebola outbreak in rural Sierra Leone.
Setting: All 100 health facilities providing maternal services in Moyamba, Sierra Leone, a rural district that experienced a smaller Ebola outbreak than other areas. Objective: To compare trends in antenatal care (the first and fourth visit [ANC1 and ANC4]), delivery, and postnatal care (PNC1) service utilisation before, during and after the Ebola outbreak (2014-2016). Design: Cross-sectional study using secondary programme data. Results: A total of 211 Ebola cases occurred in Moyamba District. The mean number of monthly ANC visits remained stable over time, except for the subset of care provided via outreach visits where, compared with before the outbreak (n = 390), ANC1 visits declined during (n = 331, P = 0.002) and after the outbreak (n = 342, P = 0.03). Most (>97%) deliveries occurred in health facilities, assisted by maternal and child health aides (>80%). During the outbreak, the mean number of community-based deliveries per month declined from 31 to 21 (P = 0.03), and the mean number of deliveries performed by midwives increased from 49 to 78 (P < 0.001) compared with before the outbreak. Before, during and after Ebola, there was no significant change in the mean number of live births (respectively n = 1134, n = 1110, n = 1162), maternal PNC1 (respectively n = 1110, n = 1105, n = 1165) or neonatal PNC1 (respectively n = 1028, n = 1050, n = 1085). Conclusion: In a rural district less affected by Ebola transmission than other areas, utilisation of maternal primary care remained robust, despite the outbreak.


Active and passive Ca2+ fluxes across cell membranes of the guinea-pig taenia coli.
The exchange of Ca between the extracellular fluid and the cellular compartment has been investigated in smooth muscle cells of taenia coli. It was found that during the initial phase of metabolic depletion by DNP + IAA, the net inwards flux of Ca amounts to 0.02 pmol cm(-2)-sec(-1). This increase might be proportional to the physiological calcium leak. The study of the relation between the inwardly directed Na gradient and the cellular Ca content has revealed that this Na gradient exerts no effect during prolonged exposure to K-free solution and a very limited effect during exposure to Na-deficient solutions. The cellular 45Ca release induced by metabolic inhibition is not affected by substituting Li or choline for Na. The supplementary calcium which enters the cells during exposure to a solution at low temperature is extruded on returning to a solution at 35 degrees C, even if the Na gradient is reversed. This finding and the effects of metabolic inhibition indicate that Ca extrusion in smooth muscle cells is a process which depends on metabolism and which is not affected by the inwardly directed Na gradient.


[Extension of life from a psychogeriatric viewpoint].
Life prolongation (lp) is not only a quantitative, but also a qualitative concept, especially when we look at the factors, which cause life prolongation. One of the consequences of lp is that in the Western world most people live into old age (defined as reaching the age of 65). Now the question is: do we have to continue to work on this product of 'progression-mindedness'. Do we like lp, considering all the negative features which accompany this phenomenon? We think that to a certain extent quality of life has been affected by lp. Qualitative lp we connect to the meaningfulness of lp. What do we understand by meaning? First, we describe some cases. These cases show that meaning is an interactive process, which, we can best conceptualize as commitment. We describe the different categories of commitment and their function. This makes clear the relations between commitment and the development of the individual older person. A consequence would be, that where commitments no longer exist, life prolongation should not be pursued.


[Successions in the forest-steppe under climate changes: a modelling approach].
The paper represents an attempt to apply the general principles of modelling vegetation dynamics under climate changes to a study of the long-term vegetation dynamics in the forest-steppe zone of the European territory of Russia, with a purpose to forecast under special climatic scenarios. An original technique is used to construct a Markov chain as a model of vegetation succession. The technique emanated from gebotanic knowledge generalized as a scheme of successional transitions with estimates of the average duration for certain stages of succession. Whenever the knowledge related the stage duration to certain (climate-sensitive) factors of the environment, the fundamental potentiality arises to model the temporal course of succession as a function of a given scenario for how the key factors change. In the formal terms, the model represents a random chain of the Markov kind with a finite number of states and discrete time of transitions by the given scheme. Relative square distributions of succession stages under concern at any time moment (within an adopted scenario) appear as the model outcome (forecasts), as well as estimates of the attainment time for certain states of the vegetation in the territory under study. A method is proposed to describe dynamics of the phytomass production and stores (and the corresponding model trajectories are obtained) for a given scenario.


Ultra high-performance liquid chromatography-tandem mass spectrometry for the simultaneous analysis of asparagine, sugars, and acrylamide in Maillard reactions.
We developed an automated microwave digestion labstation (MDL) combined with ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method under the control of positive-negative ion switching as a robust kinetic study tool for rapid and simultaneous quantification of asparagine, glucose, fructose, and acrylamide in Maillard reaction products. Maillard reactions were conducted in a potato model via MDL. The two-step simple pretreatment procedures included the addition of isotope internal standards (15)N(2)-asparagine, (13)C(6)-glucose, and D(3)-acrylamide, followed by appropriate dilution with the mobile phase and filtration. Analytes were separated on a Hypercarb column and monitored by MS/MS. Study of matrix effects indicated Maillard reaction products induce an ionization suppression of both positive and negative precursor ions, but quantitative results are corrected through the use of isotopically labeled internal standards. Using this method, the limit of detection (LOD) and limit of quantification (LOQ) ranges of all analytes were calculated as 0.04-0.6 and 0.1-1.1 μM, respectively. Excellent repeatability (RSD < 9.6%) and acceptable within-laboratory reproducibility (RSD < 9.2%) substantially supported the use of this method for sample analysis. The present kinetic tools, with 10-50 min mimic of Maillard reactions and short instrumental run time (5.5 min per sample), were successfully validated and applied to simultaneous determination of acrylamide and its precursors and intermediates during Maillard reactions and kinetic elucidation. Furthermore, current tools of MDL combined with simple sample treatment procedures and UHPLC-MS/MS analysis reduce sample analysis time and labor in the kinetic study.


Development of a Tailored HIV Prevention Intervention for Single Young Men Who Have Sex With Men Who Meet Partners Online: Protocol for the myDEx Project.
New cases of human immunodeficiency virus (HIV) among young men who have sex with men (YMSM), aged 18 to 24, underscore the importance of developmentally-informed HIV programs for YMSM. We developed an online intervention focused on risk reduction strategies across different sexual partner types. Intervention activities focus on assisting YMSM reflect on their partner-seeking behaviors, develop sexual decision-making rules to reduce their HIV risks, and consider the adoption of HIV prevention behaviors. This pilot, randomized controlled trial (RCT) aims to examine the feasibility, acceptability, and preliminary efficacy of a tailored, Web-based HIV prevention intervention for single YMSM. We designed a prospective RCT of online-recruited cis-gender men (N=180) who reported recent unprotected anal intercourse, self-report as HIV negative or are unaware of their HIV status, and meet sexual partners through online dating apps. Individuals in the control arm receive an attention-control condition that includes HIV/sexually transmitted infection (STI) information currently available on sex education websites. Individuals in the intervention arm receive a 6-session Web-based program tailored on their demographic information, partner-seeking behaviors and relationship desires, and prior sexual attitudes and behaviors. This tailored content will match HIV prevention messages and safer sex skills with YMSM's outcome expectancies when meeting new partners and thereby help them consider how to integrate safer sex practices into different partner types. Study assessments are taken at baseline, 30-, 60-, and 90-day follow-ups. Intervention acceptability and preliminary efficacy will be explored in sexual risk behaviors and HIV/STI testing. The RCT launched in November 2016 and is ongoing. To date, 180 eligible individuals have been enrolled, consented, and randomized. Of the 120 individuals in the intervention arm, 51.7% (62/120) identify as non-Hispanic white and half of the control arm identifies as non-Hispanic white. There were no differences observed by arm for race and/or ethnicity, age, or sexual orientation. Although there are in-person evidence-based interventions with proven efficacy for YMSM, few HIV/STI prevention interventions delivered online exist. Online interventions may ease access to comprehensive HIV/STI education among YMSM and allow personalized content to be delivered. The online intervention that we developed, myDEx, aims to alleviate the gaps within HIV prevention for YMSM by utilizing tailored, Web-based content with the goal of developing skills for same-sex dating and relationship building, while reducing their risks for HIV/STI. ClinicalTrials.gov NCT02842060; https://clinicaltrials.gov/ct2/show/NCT02842060 (Archived by WebCite at http://www.webcitation.org/6rcJdxF9v).


Vascular endothelial growth factor expression in serum of patients with hepatocellular carcinoma.
To determine the pre-therapeutic serum level of vascular endothelial growth factor (VEGF) in patients with hepatocellular carcinoma (HCC) and to elucidate the relation between the serum level and clinical characteristics and metastasis of HCC. One-hundred and fifteen HCC patients, 40 patients with benign liver lesions, and 30 healthy control subjects were included in this study. The serum VEGF level was measured with the quantitative sandwich enzyme linked immunosorbent assay (ELISA, R&D systems). The serum VEGF levels in the HCC group (465.62 +/- 336.24 pg/ml) was significantly elevated as compared with those in patients with benign liver lesions (159.54 +/- 120.58 pg/ml) and those in normal controls (123.53 +/- 51.84 pg/ml). The VEGF levels were not significantly different between the patients with benign liver lesions and the normal controls. The serum VEGF levels showed a positive rate of 77.4%, 25%, and 3.3% in the HCC patients, benign liver lesion patients and normal controls, respectively. In the 115 HCC patients, the serum VEGF levels in patients with portal vein (PV) emboli (n = 26, 582.76 +/- 441.89 pg/ml), with metastasis (n = 43, 548.29 +/- 438.57 pg/ml) or with large HCC lesions (>/= 5 cm in diameter) (n = 69, 554.43 +/- 369.99 pg/ml) were significantly higher than those without PV-emboli (n = 89, 431.39 +/- 292.84 pg/ml), without metastasis (n = 72, 416.24 +/- 247.27 pg/ml) or with small HCC lesions (n = 42, 328.67 +/- 227.47 pg/ml). The serum VEGF levels in stage I, II, III, IVa and IVb HCC patients were 340.6 pg/ml, 451.55 +/- 307.84 pg/ml, 397.44 +/- 257.18 pg/ml, 486.10 +/- 397.73 pg/ml and 647.93 +/- 344.56 pg/ml, respectively. The pre-therapeutic serum VEGF levels in HCC patients appear to reflect the disease's potential activity of vascular invasion and metastasis.


Knowledge and attitudes toward AIDS among female college students in Nagasaki, Japan.
This study assesses knowledge and attitudes concerning HIV infection and individuals with AIDS among 383 female students attending colleges in Nagasaki, Japan. A structured questionnaire containing questions concerning knowledge about AIDS, sources of information, beliefs and attitudes toward people with HIV/AIDS was administered during sessions set up for that purpose. The mean age of participants was 18.8 +/- 0.8 years (+/- SD). The main source of information for AIDS awareness as reported by the students was the mass media. Good knowledge about AIDS was positively associated with ease of acceptance of living in the same house with a person diagnosed with AIDS [odds ratio (OR): 1.90; 95% confidence interval (CI): 1.07-3.38]. However, residing at home (OR: 0.64; 95% CI: 0.42-0.98) and involvement in nurse education programmes (OR: 0.59; 95% CI: 0.37-0.95) showed a negative association. Students demonstrated a high level of knowledge concerning AIDS and HIV, but had considerable misconceptions and prejudices about people having HIV/AIDS. Our results suggest that a more appropriate education programme in colleges in Japan may be necessary to reduce the discrepancy between general knowledge and desirable attitude regarding HIV/AIDS.


Dipole-switch induced modification of the emissive response of carbon nanotubes.
The interaction of carbon nanotubes with the molecular dipole switch spiropyran is expected to affect the optical response of the tubes. Until now, the need of anchor groups to immobilize the switches on the tubes has hindered the experimental observation of the effects of switching on the emission behavior of the tubes. Here we present spiropyran-carbon nanotube complexes obtained by micelle swelling. This method does not require any anchor nor sophisticated chemistry to warrant close tube-switch proximity. For the first time, we observe the shifts predicted theoretically and their effect on the tubes' excitation and emission energies.


Thermal optimisation of the Reimer-Tiemann reaction using thermochromic liquid crystals on a microfluidic reactor.
Microreactors incorporating thin film resistive heating elements for continuous flow organic synthesis are presented. Internal thermal conditions were monitored in real time using reflectance spectra of temperature sensitive thermochromic liquid crystals (TLC) in a collateral microfluidic network. To demonstrate the precise temperature control provided by this method, the thermal optimisation of the Reimer-Tiemann formylation of beta-naphthol was performed under hydrodynamic pumping regimes.


Update on hypersomnias of central origin.
To describe the multiple clinical aspects of hypersomnias of central origin. Emphasis is given to the new pathophysiological pathways and treatment options described in the current literature. Narcolepsy is the most recognized of the hypersomnias of central origin. Hypocretin deficiency appears to underlie narcolepsy with cataplexy, and infections and vaccinations have been associated with disease onset. Targeted therapeutic approaches are currently underway. A putative naturally occurring constituent in the cerebrospinal fluid of patients with non-narcoleptic primary hypersomnias, able to stimulate γ-aminobutyric acid alpha receptors and induce sleep, has recently been postulated. Neuroimaging has also provided more insight into the pathophysiology of Kleine-Levin syndrome. Sleep deprivation is currently recognized as a major differential diagnosis. Excessive daytime sleepiness is the cardinal symptom of the hypersomnias of central origin, with major impact on the quality of life. It is important that clinicians be able to recognize these conditions, so that appropriate management or onward referral is expedited.


Time to Spread Your Wings: A Review of the Avian Ancient DNA Field.
Ancient DNA (aDNA) has the ability to inform the evolutionary history of both extant and extinct taxa; however, the use of aDNA in the study of avian evolution is lacking in comparison to other vertebrates, despite birds being one of the most species-rich vertebrate classes. Here, we review the field of "avian ancient DNA" by summarising the past three decades of literature on this topic. Most studies over this time have used avian aDNA to reconstruct phylogenetic relationships and clarify taxonomy based on the sequencing of a few mitochondrial loci, but recent studies are moving toward using a comparative genomics approach to address developmental and functional questions. Applying aDNA analysis with more practical outcomes in mind (such as managing conservation) is another increasingly popular trend among studies that utilise avian aDNA, but the majority of these have yet to influence management policy. We find that while there have been advances in extracting aDNA from a variety of avian substrates including eggshell, feathers, and coprolites, there is a bias in the temporal focus; the majority of the ca. 150 studies reviewed here obtained aDNA from late Holocene (100-1000 yBP) material, with few studies investigating Pleistocene-aged material. In addition, we identify and discuss several other issues within the field that require future attention. With more than one quarter of Holocene bird extinctions occurring in the last several hundred years, it is more important than ever to understand the mechanisms driving the evolution and extinction of bird species through the use of aDNA.


Clinical Significance of Community- and Healthcare-Acquired Carbapenem-Resistant Enterobacteriaceae Isolates.
This study was conducted to investigate the clinical significance, manifestations, microbiological characteristics and outcomes of carbapenem-resistant Enterobacteriaceae (CRE) isolates, and compare the clinical features of community- and healthcare-acquired CRE isolates. A total of 78 patients were identified to have CRE. Klebsiella pneumoniae was the most common pathogens (n = 42, 53.8%), followed by Enterobacter cloacae (n = 24, 30.8%), and Escherichia coli (n = 11, 14.1%). Most of the patients acquired CRE from healthcare settings (n = 55, 70.5%), and other cases got CRE from community settings (n = 23, 29.5%). Nine cases (11.5%) were classified as CRE colonization. Among the remaining 69 cases of CRE infections, pneumonia (n = 28, 40.6%) was the most common type of infections, followed by urinary tract infection (n = 24, 34.8%), and intra-abdominal infection (n = 16, 23.2%). The patients acquired CRE from community settings were more likely to be elderly, female, and had more urinary tract infections than from healthcare settings. In contrast, the patients acquired CRE from healthcare settings had more intra-abdominal infections, intra-abdominal surgery, and presence of indwelling device than from community settings. In conclusion, community-acquired CRE are not rare, and their associated clinical presentations are different from healthcare-acquired CRE.


Safety of a Brief Emergency Department Observation Protocol for Patients With Presumed Fentanyl Overdose.
Fentanyl overdoses are increasing and few data guide emergency department (ED) management. We evaluate the safety of an ED protocol for patients with presumed fentanyl overdose. At an urban ED, we used administrative data and explicit chart review to identify and describe consecutive patients with uncomplicated presumed fentanyl overdose (no concurrent acute medical issues) from September to December 2016. We linked regional ED and provincial vital statistics databases to ascertain admissions, revisits, and mortality. Primary outcome was a composite of admission and death within 24 hours. Other outcomes included treatment with additional ED naloxone, development of a new medical issue while in the ED, and length of stay. A prespecified subgroup analysis assessed low-risk patients with normal triage vital signs. There were 1,009 uncomplicated presumed fentanyl overdose, mainly by injection. Median age was 34 years, 85% were men, and 82% received out-of-hospital naloxone. One patient was hospitalized and one discharged patient died within 24 hours (combined outcome 0.2%; 95% confidence interval [CI] 0.04% to 0.8%). Sixteen patients received additional ED naloxone (1.6%; 95% CI 1.0% to 2.6%), none developed a new medical issue (0%; 95% CI 0% to 0.5%), and median length of stay was 173 minutes (interquartile range 101 to 267). For 752 low-risk patients, no patients were admitted or developed a new issue, and one died postdischarge; 3 (0.4%; 95% CI 0.01% to 1.3%) received ED naloxone. In our cohort of ED patients with uncomplicated presumed fentanyl overdose-typically after injection-deterioration, admission, mortality, and postdischarge complications appear low; the majority can be discharged after brief observation. Patients with normal triage vital signs are unlikely to require ED naloxone.


Tenacibaculum sediminilitoris sp. nov., isolated from a tidal flat.
A Gram-stain-negative, non-flagellated, non-gliding, non-spore-forming bacterial strain, designated YKTF-3T, was isolated from a tidal flat on the Yellow Sea, South Korea. Strain YKTF-3T grew optimally at 30 °C and in the presence of 2.0-4.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain YKTF-3T fell within the clade comprising the type strains of Tenacibaculumspecies. The novel strain exhibited 16S rRNA gene sequence similarity values of 97.22-98.34 % to the type strains of Tenacibaculum gallaicum, Tenacibaculum ascidiaceicola, Tenacibaculum litoreum, Tenacibaculum discolor, Tenacibaculum aestuarii and Tenacibaculum lutimaris, and of 95.08-96.95 % to the type strains of the other Tenacibaculum species. Strain YKTF-3T contained MK-6 as the predominant menaquinone and iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and iso-C17 : 0 3-OH as the major fatty acids. The major polar lipids of strain YKTF-3T were phosphatidylethanolamine, one unidentified lipid, one unidentified aminophospholipid, one unidentified glycolipid and one unidentified aminolipid. The DNA G+C content was 32.3 mol% and its mean DNA-DNA relatedness values with the type strains of the six phylogenetically closely relatedTenacibaculum species were 10-27 %. Differential phenotypic properties, together with phylogenetic and genetic distinctiveness, revealed that strain YKTF-3T is separate from other recognized species of the genus Tenacibaculum. On the basis of the data presented, strain YKTF-3T is considered to represent a novel species of the genus Tenacibaculum, for which the name Tenacibaculum sediminilitoris sp. nov. is proposed. The type strain is YKTF-3T (=KCTC 52210T=NBRC 111991T).


Compensation for decreased expression of B7 molecules on Leishmania infantum-infected canine macrophages results in restoration of parasite-specific T-cell proliferation and gamma interferon production.
Infection of humans and dogs by Leishmania infantum may result in visceral leishmaniasis, which is characterized by impaired T-cell-mediated immune responses to parasite antigens. Dogs are natural hosts of Leishmania parasites and play an important role in the transmission of the parasites to humans. In an effort to characterize the immune response in dogs infected with this intracellular pathogen, we examined how infection with L. infantum affects canine macrophages and the consequences for T-cell activation in vitro. We showed that the proliferation of T-cell lines to cognate antigen decreases to background levels when infected autologous monocyte-derived macrophages are used as antigen-presenting cells (APC). The observed reduction of antigen-specific T-cell proliferation was shown to be dependent on the parasite load and to require cell-to-cell interaction of T cells with the infected APC. In addition, we observed a decreased expression of costimulatory B7 molecules on infected monocyte-derived macrophages. The expression of other surface molecules involved in T-cell activation, such as major histocompatibility complex class I and class II, on these cells did not change upon infection, whereas the expression of intracellular adhesion molecule 1 was marginally increased. Compensation for the decreased expression of B7 molecules by the addition of B7-transfected cells resulted in the restoration of cell proliferation and gamma interferon (IFN-gamma) production by a Leishmania-specific T-cell line. These results showed that for the activation of parasite-specific canine T cells producing IFN-gamma, which are most likely involved in protective immunity, sufficient expression of B7 molecules on infected macrophages is required. Provision of costimulatory molecules may be an approach for immunotherapy of leishmaniaisis as well as for vaccine development.


One-year treatment with 0.1% tacrolimus ointment versus a corticosteroid regimen in adults with moderate to severe atopic dermatitis: A randomized, double-blind, comparative trial.
A one-year, randomized, double-blind study was conducted in 80 patients with atopic dermatitis treated with tacrolimus ointment or a corticosteroid regimen (hydrocortisone acetate 1% ointment for head and neck, hydrocortisone butyrate 0.1% ointment for trunk and limbs) to compare efficacy and safety, and effects on Th2-reactivity. The study was completed by 36/40 patients in the tacrolimus group, and 31/40 patients in the corticosteroid group. In both groups affected body surface area, eczema area and severity index, and transepidermal water loss decreased at months 6 and 12. Tacrolimus was superior for all efficacy scores at month 6, and in the head and neck area at month 12. Recall antigen reactivity increased at month 12 in both groups. Adverse events were reported by 40/40 patients in the tacrolimus, and by 34/40 patients in the corticosteroid group. Long-term treatment with topical tacrolimus or a corticosteroid regimen improves atopic dermatitis and recall antigen reactivity, suggesting an improvement in the Th1/Th2-balance.


Exposure to genotoxic compounds alters in vitro cellular VOC excretion.
Genotoxic carcinogens significantly damage cells and tissues by targeting macromolecules such as proteins and DNA, but their mechanisms of action and effects on human health are diverse. Consequently, determining the amount of exposure to a carcinogen and its cellular effects is essential, yet difficult. The aim of this manuscript was to investigate the potential of detecting alterations in volatile organic compounds (VOCs) profiles in the in vitro headspace of pulmonary cells after exposure to the genotoxic carcinogens cisplatin and benzo[a]pyrene using two different sampling set-ups. A prototype set-up was used for the cisplatin exposure, whereas a modified set-up was utilized for the benzo[a]pyrene exposure. Both carcinogens were added to the cell medium for 24 h. The headspace in the culture flask was sampled to measure the VOC content using gas chromatography-time-of-flight-mass spectrometry. Eight cisplatin-specific VOCs and six benzo[a]pyrene-specific VOCs were discriminatory between treated and non-treated cells. Since the in vivo biological effects of both genotoxic compounds are well-defined, the origin of the identified VOCs could potentially be traced back to common cellular processes including cell cycle pathways, DNA damage and repair. These results indicate that exposing lung cells to genotoxins alters headspace VOC profiles, suggesting that it might be possible to monitor VOC changes in vivo to study drug efficacy or exposure to different pollutants. In conclusion, this study emphasizes the innovative potential of in vitro VOCs experiments to determine their in vivo applicability and discover their endogenous origin.


Ultrasound-guided fine-needle aspiration biopsy in nonpalpable thyroid nodules: is it useful in infracentimetric nodules?
The purpose of this study was to evaluate the usefulness of an ultrasound-guided fine-needle aspiration biopsy (US-FNAB) for the evaluation and treatment planning of nonpalpable thyroid lesions, including infracentimetric nodules. One hundred and twenty one patients underwent US-FNAB for 149 non-palpable solid nodules. Sixty-five patients underwent surgery, and 84 were followed up for at least 36 months. The results of the US-FNAB correlated with the pathological findings and clinical follow-up results. The nodules ranged from 0.3 to 2 cm in diameter, with a mean of 0.8 cm. Among the 149 nodules, 115 were infracentimetric and 34 were centimetric or supracentimetric in size. Of the 149 thyroid nodules, US-FNAB was true positive in 43, true negative in 90, false positive in 7 and false negative in 1. In 8 cases, the lesion was inadequately sampled. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of the US-FNAB for the infracentimetric nodules were 96.9, 93.4, 86.1, 98.6 and 94.4%, and for the centimetric or supracentimetric nodules, were 100, 90.5, 85.7, 100 and 93.9%, respectively. There were no significant differences in the results between the infracentimetric and centimetric or supracentimetric nodules. In conclusion, an US-FNAB is a useful tool for determining the treatment plan of non-palpable solid thyroid nodules, even when less than 1 cm in diameter, and shows high sensitivity, specificity and accuracy.


Identification of potent inhibitors of SARS-CoV-2 infection by combined pharmacological evaluation and cellular network prioritization.
Pharmacologically active compounds with known biological targets were evaluated for inhibition of SARS-CoV-2 infection in cell and tissue models to help identify potent classes of active small molecules and to better understand host-virus interactions. We evaluated 6,710 clinical and preclinical compounds targeting 2,183 host proteins by immunocytofluorescence-based screening to identify SARS-CoV-2 infection inhibitors. Computationally integrating relationships between small molecule structure, dose-response antiviral activity, host target, and cell interactome produced cellular networks important for infection. This analysis revealed 389 small molecules with micromolar to low nanomolar activities, representing >12 scaffold classes and 813 host targets. Representatives were evaluated for mechanism of action in stable and primary human cell models with SARS-CoV-2 variants and MERS-CoV. One promising candidate, obatoclax, significantly reduced SARS-CoV-2 viral lung load in mice. Ultimately, this work establishes a rigorous approach for future pharmacological and computational identification of host factor dependencies and treatments for viral diseases.


Screening for consequences of trauma - an update on the global collaboration on traumatic stress.
This letter provides an update on the activities of "The Global Collaboration on Traumatic Stress" (GC-TS) as first described by Schnyder et al. in 2017. It presents in further detail the projects of the first theme, in particular the development of and initial data on the Global Psychotrauma Screen (GPS), a brief instrument designed to screen for the wide range of potential outcomes of trauma. English language data and ongoing studies in several languages provide a first indication that the GPS is a feasible, reliable and valid tool, a tool that may be very useful in the current pandemic of the coronavirus disease 2019 (COVID-19). Further multi-language and cross-cultural validation is needed. Since the start of the GC-TS, new themes have been introduced to focus on in the coming years: a) Forcibly displaced persons, b) Global prevalence of stress and trauma related disorders, c) Socio-emotional development across cultures, and d) Collaborating to make traumatic stress research data "FAIR". The most recent theme added is that of Global crises, currently focusing on COVID-19-related projects.


Sustained in vivo cardiac protection by a rationally designed peptide that causes epsilon protein kinase C translocation.
Brief periods of cardiac ischemia trigger protection from subsequent prolonged ischemia (preconditioning). epsilon Protein kinase C (epsilonPKC) has been suggested to mediate preconditioning. Here, we describe an epsilonPKC-selective agonist octapeptide, psiepsilon receptor for activated C-kinase (psiepsilonRACK), derived from an epsilonPKC sequence homologous to its anchoring protein, epsilonRACK. Introduction of psiepsilonRACK into isolated cardiomyocytes, or its postnatal expression as a transgene in mouse hearts, increased epsilonPKC translocation and caused cardio-protection from ischemia without any deleterious effects. Our data demonstrate that epsilonPKC activation is required for protection from ischemic insult and suggest that small molecules that mimic this epsilonPKC agonist octapeptide provide a powerful therapeutic approach to protect hearts at risk for ischemia.


Puberty-dependent sleep regulation and alcohol use in early adolescents.
Research has shown a bi-directional relation between alcohol use and sleep regulation in adults. Much less is known about this association in early adolescents, while profound puberty-dependent transitions regarding sleep patterns take place in early adolescence. Moreover, puberty has been associated with an increase in alcohol use of adolescents. In this study, we investigated the associations between pubertal development, sleep preference, sleep problems, and alcohol use in 431 early adolescents (mean age: 13.66). Second, it was studied whether the associations changed when controlling for adolescent internalizing and externalizing problems. Furthermore, we included gender as a moderator on all the associations. Results showed that pubertal development was positively associated with sleep problems and more evening-type tendencies (e.g., favoring later bedtimes), which in turn were positively related to alcohol use. Underlying psychopathology, gender and educational level did not change these relationships. From this study, it can be concluded that both puberty and sleep regulation are important factors in explaining alcohol use in early adolescence.


Similarity of strain- and route-dependent murine responses to an adenovirus vector using the homologous thrombopoietin cDNA as the reporter genes.
Replication-deficient adenovirus (Ad) vectors are effective in transferring genes in vivo, but their use is associated with significant variation in the extent and/or duration of expression observed among different strains of experimental animals and different routes of administration of the vector. We have minimized the variables of the heterologous transgene and animal-to-animal variation by constructing an Ad vector encoding murine thrombopoietin (mTPO, AdmTPO), a homologous protein that induces a physiologic response (elevation of blood platelet levels) that can be followed sequentially over time in the same animal. Using the C57BL/6 and BALB/c stains, liver administration was accomplished by intravenous administration and skeletal muscle administration by direct injection. Despite the use of a homologous cDNA as a transgene, the Ad genome was rapidly lost from the liver after intravenous administration over the first 1 to 2 weeks, with no difference in pattern of decline between the C57BL/6 and BALB/c strains. Both strains exhibited a cytotoxic T lymphocyte (CTL) response directed against the AdmTPO vector. Consistent with the decline in vector genome over time, the initial high levels of mTPO mRNA in the liver declined to an undetectable level within 2 weeks. Platelet counts peaked at 8- to 10-fold above baseline within the first 2 weeks, and then gradually declined, returning to normal level by 50 to 60 days. Intravenous administration of the AdmTPO vector to beta2-microglobulin-deficient mice resulted in a longer persistence of elevated platelets levels, although the eventual return of platelet levels to normal in these mice suggests the elimination of the Ad vector cannot be explained solely by CTL response. Although the intramuscular administration of the AdmTPO vector resulted in platelet levels with a lower peak and minor differences over time compared with the intravenous route, the C57BL/6 and BALB/c strains demonstrated the same rapid loss of Ad genome and mTPO mRNA levels in the muscle as in the liver. Together, these observations suggest that simplifying the experimental design by eliminating the variable of host response to a heterologous transgene, and by following the consequences of gene transfer in the same animals over time, there can be remarkable similarity in strain- and route-dependent responses to an Ad vector.


Microtensile bond strength of resin cement to a feldspathic ceramic.
The purpose of this study was to evaluate the microtensile bond strength of resin cement to a feldspathic ceramic after treating the surface with (a) hydrofluoric (HF) acid, (b) air abrasion, (c) Er:YAG laser irradiation, (d) Nd:YAG laser irradiation, and (e) HF acid etching after either air abrasion or laser irradiation. It is unknown whether the laser application or its combination with another treatment method can be used as a tool to roughen the surface of a feldspathic ceramic in order to increase the bond strength between the resin cement and ceramic surface. Forty feldspathic ceramic blocks (Ceramco(TM)) were prepared and divided into eight equal groups (n = 5) according to the following surface treatments: no treatment; etching with 9.5% HF acid; air abrasion with 50 μm Al(2)O(3); Er:YAG laser irradiation; Nd:YAG laser irradiation; air abrasion plus acid etching; Er:YAG laser plus acid etching; and Nd:YAG laser plus acid etching. After surface treatment, a silane-coupling agent and resin cement (Panavia F(TM)) were applied to each block. After storing for 24 h at 37°C and thermocycling between 5°C and 55°C for 1000 cycles, the microtensile bond strength of each specimen was measured. The highest bond strength was obtained from HF acid etching. HF acid etching after each laser irradiation significantly increased the bond strength (p < 0.05). However, HF acid etching after air abrasion decreased bond strength when compared to air abrasion alone. HF acid etching is the most effective surface treatment method for a feldspathic ceramic. However, laser irradiation with either the Er:YAG or Nd:YAG laser is not an adequate method for improving the bond strength of Panavia F. The laser application should be combined with HF acid etching.


Spartin-mediated lipid transfer facilitates lipid droplet turnover.
Lipid droplets (LDs) are organelles critical for energy storage and membrane lipid homeostasis, whose number and size are carefully regulated in response to cellular conditions. The molecular mechanisms underlying lipid droplet biogenesis and degradation, however, are not well understood. The Troyer syndrome protein spartin (SPG20) supports LD delivery to autophagosomes for turnover via lipophagy. Here, we characterize spartin as a lipid transfer protein whose transfer ability is required for LD degradation. Spartin co-purifies with phospholipids and neutral lipids from cells and transfers phospholipids in vitro via its senescence domain. A senescence domain truncation that impairs lipid transfer in vitro also impairs LD turnover in cells while not affecting spartin association with either LDs or autophagosomes, supporting that spartin's lipid transfer ability is physiologically relevant. Our data indicate a role for spartin-mediated lipid transfer in LD turnover. The Troyer syndrome protein spartin was proposed to function as a lipophagy receptor that delivers lipid droplets, organelles key for energy storage and membrane lipid homeostasis, to autophagosomes for degradation. We identify an additional function for spartin as a lipid transfer protein and show its transfer ability is required for lipid droplet degradation, including by lipophagy. Our data support that protein-mediated lipid transfer plays a role in lipid droplet turnover. Moreover, in spartin's senescence domain we have discovered a new lipid transport module that likely also features in still undiscovered aspects of lipid droplet biology and membrane homeostasis.


Exceptional plant penetration and feeding upon cortical parenchyma cells by the woolly poplar aphid.
Forty percent of aphids live wholly or partly on trees, most species being associated with leaves or petioles. Species able to exploit woody parts have either specific adaptations, such as extra long stylets that allow them to reach the phloem, or the ability to induce galls. The woolly poplar aphid, Phloeomyzus passerinii (Signoret) (Hemiptera: Aphididae), colonizes the trunks and base of the lower branches of mature poplars and causes cortical necrosis leading to the death of trees where infestation is heavy. Very little is known about the mode of feeding of P. passerinii. This study looked at the feeding behavior of P. passerinii on stem-cuttings of Populus x canadensis Moench using: (i) histological analyses of the feeding site and stylet pathway and (ii) electrical penetration graphs (EPG, DC) based on parthenogenetic apterous females on woody tissues. The histological and EPG results showed that stylets of P. passerinii penetrated into the plant tissues following a straight unbranched extracellular and intracellular pathway to reach the cortical parenchyma. Compared to EPGs for phloem sap feeding aphids, there were differences in the waveforms A and C whereas a new waveform Icp was described. Based on histological analyses and previous descriptions of EPG waveforms, correlations with the stylet tip position and aphid activities within bark tissues are discussed. A pathway and a sustained intracellular phase were distinguished, both occurring in the cortical parenchyma cells. The bark aphid feeding mode is discussed in relation to the damage caused and in terms of changes in the aphid's diet.


Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.
miR-206, a member of the so-called myomiR family, is largely acknowledged as a specific, positive regulator of skeletal muscle differentiation. A growing body of evidence also suggests a tumor suppressor function for miR-206, as it is frequently downregulated in various types of cancers. In this study, we show that miR-206 directly targets cyclin D1 and contributes to the regulation of CCND1 gene expression in both myogenic and non-muscle, transformed cells. We demonstrate that miR-206, either exogenous or endogenous, reduces cyclin D1 levels and proliferation rate in C2C12 cells without promoting differentiation, and that miR-206 knockdown in terminally differentiated C2C12 cells leads to cyclin D1 accumulation in myotubes, indicating that miR-206 might be involved in the maintenance of the post-mitotic state. Targeting of cyclin D1 might also account, at least in part, for the tumor-suppressor activity suggested for miR-206 in previous studies. Accordingly, the analysis of neoplastic and matched normal lung tissues reveals that miR-206 downregulation in lung tumors correlates, in most cases, with higher cyclin D1 levels. Moreover, gain-of-function experiments with cancer-derived cell lines and with in vitro transformed cells indicate that miR-206-mediated cyclin D1 repression is directly coupled to growth inhibition. Altogether, our data highlight a novel activity for miR-206 in skeletal muscle differentiation and identify cyclin D1 as a major target that further strengthens the tumor suppressor function proposed for miR-206.


In-situ reconstruction of rock-like 3D hierarchical MIL-53(Fe) self-supporting electrode with oxygen vacancy induced ultra-long stable and efficient water oxidation.
The exploitation of efficient, stable and cheap electrocatalyst for oxygen evolution reaction (OER) is very significant to the development of energy technology. In this study, Fe-based metal-organic frameworks (MIL-53(Fe)) self-supporting electrode with a 3D hierarchical open structure was developed through a semi-sacrificial strategy. The self-supporting electrode exhibits an excellent OER performance with an overpotential of 328 mV at 100 mA cm-2 in 1 M KOH, which is superior than that of IrO2 catalyst. Importantly, the optimized self-supporting electrode could operate at 100 mA cm-2 for 520 h without visible decrease in activity. It was also found that the structure of MIL-53(Fe) was in-situ self-reconstructed into oxyhydroxides during OER process. However, the 3D hierarchical open structure assembled with nano-microstructures kept well, which ensured the long-term stability of our self-supporting electrode for OER. Furthermore, density functional theory (DFT) calculations reveal that the FeOOH with rich oxygen vacancy transformed from MIL-53(Fe) plays a key role for the OER catalytic activity. And, the uninterrupted formation of oxygen vacancy during OER process ensures the continuous OER catalytic activity, which is the original source for the ultra-long stability of the self-supporting electrode toward OER. This work explores the way for the construction of efficient self-supporting oxygen electrodes based on MOFs.


Synaptic overflow of dopamine in the nucleus accumbens arises from neuronal activity in the ventral tegmental area.
Dopamine concentrations fluctuate on a subsecond time scale in the nucleus accumbens (NAc) of awake rats. These transients occur in resting animals, are more frequent following administration of drugs of abuse, and become time-locked to cues predicting reward. Despite their importance in various behaviors, the origin of these signals has not been demonstrated. Here we show that dopamine transients are evoked by neural activity in the ventral tegmental area (VTA), a brain region containing dopaminergic cell bodies. The frequency of naturally occurring dopamine transients in a resting, awake animal was reduced by a local VTA microinfusion of either lidocaine or (+/-)2-amino,5-phosphopentanoic acid (AP-5), an NMDA receptor antagonist that attenuates phasic firing. When dopamine increases were pharmacologically evoked by noncontingent administration of cocaine, intra-VTA infusion of lidocaine or AP-5 significantly diminished this effect. Dopamine transients acquired in response to a cue during intracranial self-stimulation were also attenuated by intra-VTA microinfusion of AP-5, and this was accompanied by an increase in latency to lever press. The results from these three distinct experiments directly demonstrate, for the first time, how neuronal firing of dopamine neurons originating in the VTA translates into synaptic overflow in a key terminal region, the NAc shell.


[Fluctuations and dyskinesias as early L-dopa-induced motor complications in severe Parkinsonian's patients].
Daily fluctuations of motor performance and dyskinesias in patients with Parkinson's disease (PD) treated with levodopa represent a difficult challenge to our understanding. We report 10 patients diagnosed of severe PD (Hoehn and Yahr: III-IV/V) treated with levodopa (range of dose: 750-900 mg/day) in single drug therapy since their diagnosis (mean time of levodopatherapy: 4.8 2.4 months, range: 3-6 months). All patients developed motor complications within weeks to months after initiating L-dopatherapy. Two patients received an intravenous apomorphine infusion (mean dose: 8.5 mg/day) during a mean time of 7.5 hours, but motor complications persisted during the infusion in spite of continuous dopaminergic stimulus. The degree of nigrostriatal damage (disease severity) seems to be a very important risk factor for the development of treatment-related motor complications.


A Naganishia in high places: functioning populations or dormant cells from the atmosphere?
Here, we review the current state of knowledge concerning high-elevation members of the extremophilic Cryptococcus albidus clade (now classified as the genus Naganishia). These fungi dominate eukaryotic microbial communities across the highest elevation, soil-like material (tephra) on volcanoes such as Llullaillaco, Socompa, and Saírecabur in the Atacama region of Chile, Argentina, and Bolivia. Recent studies indicate that Naganishia species are among the most resistant organisms to UV radiation, and a strain of N. friedmannii from Volcán Llullaillaco is the first organism that is known to grow during the extreme, diurnal freeze-thaw cycles that occur on a continuous basis at elevations above 6000 m.a.s.l. in the Atacama region. These and other extremophilic traits discussed in this review may serve a dual purpose of allowing Naganishia species to survive long-distance transport through the atmosphere and to survive the extreme conditions found at high elevations. Current evidence indicates that there are frequent dispersal events between high-elevation volcanoes of Atacama region and the Dry Valleys of Antarctica via "Rossby Wave" merging of the polar and sub-tropical jet streams. This dispersal hypothesis needs further verification, as does the hypothesis that Naganishia species are flexible "opportunitrophs" that can grow during rare periods of water (from melting snow) and nutrient availability (from Aeolian inputs) in one of the most extreme terrestrial habitats on Earth.


Time-multiplexed multi-view three-dimensional display with projector array and steering screen.
A novel time-multiplexed multi-view three-dimensional (3D) display has been implemented using a projector array to provide the image source and an angular steering-screen module to generate multiple high density horizontal views. The liquid crystal (LC)-based steering screen was specially developed to deflect light beams over a small range and operate in synchronism with the projector array with the use of a customized FPGA driver. The prototype produces vivid color 3D scenes with smooth parallax to multiple viewers. The experimental results verify the proposed multi-projection time-multiplexed multi-view 3D display method that uses a steering screen to produce dense views. Displaying both static and dynamic 3D contents is achieved in our implemented 36-view 3D display prototype. The results of crosstalk measurements are given and analyzed to evaluate the display performance.


Life span extension by glucose restriction is abrogated by methionine supplementation: Cross-talk between glucose and methionine and implication of methionine as a key regulator of life span.
Caloric restriction (CR) is known to extend life span across species; however, the molecular mechanisms are not well understood. We investigate the mechanism by which glucose restriction (GR) extends yeast replicative life span, by combining ribosome profiling and RNA-seq with microfluidic-based single-cell analysis. We discovered a cross-talk between glucose sensing and the regulation of intracellular methionine: GR down-regulated the transcription and translation of methionine biosynthetic enzymes and transporters, leading to a decreased intracellular methionine concentration; external supplementation of methionine cancels the life span extension by GR. Furthermore, genetic perturbations that decrease methionine synthesis/uptake extend life span. These observations suggest that intracellular methionine mediates the life span effects of various nutrient and genetic perturbations, and that the glucose-methionine cross-talk is a general mechanism for coordinating the nutrient status and the translation/growth of a cell. Our work also implicates proteasome as a downstream effector of the life span extension by GR.


Polyphenols can inhibit furin in vitro as a result of the reactivity of their auto-oxidation products to proteins.
Methods using fluorogenic peptide substrates have been proposed for screening of proprotein convertase (PC) inhibitors and they are attractive since they offer the advantage of being sensitive, cost-effective and susceptible to miniaturization. Several polyphenols, including epigallocatechin gallate ((-)EGCG), the main component of green tea, and quercetin, widely distributed in fruit and vegetables, however, led to false positive results when fluorogenic peptide substrates were used. Processing of genuine furin substrates was not inhibited by these polyphenols. In the present study, these discordant effects of (-)EGCG on the PC furin were studied. While quercetin can form aggregates in solution, aggregate-based promiscuous inhibition could be ruled out as underlying mechanism for (-)EGCG. Hydrogen peroxide production, from auto-oxidation, was too low to be a major factor but appeared associated to furin inhibition, suggesting a role for other auto-oxidation products. Since the instability of catechins is related to their electrophilic character, we tested the nucleophilic substance glutathione for stabilization. Indeed glutathione reduced furin inhibition and (-)EGCG binding to furin and serum albumin as shown by redox-cycling staining. Catechins, therefore, seem to form reactive compounds and this should be taken into account in screening assays. Adding glutathione to the detergent-based assay, as used in these studies to measure furin processing activity, strongly reduced inhibition by a number of polyphenols (catechins, gallic acid and quercetin), while the effect on the genuine inhibitor nona-D-arginine remained unchanged. the combined use of detergent and glutathione in the screening assay for furin inhibitors improves the predictive value.


How do physiotherapists solicit and explore patients' concerns in back pain consultations: a conversation analytic approach.
Background: Guidelines advocate that non-specific chronic low back pain (NSCLBP) be considered within a multi-dimensional bio-psychosocial (BPS) framework. This BPS approach advocates incorporating the patient's perspective as part of the treatment process. 'Agenda setting' has been introduced as the key to understanding patients' concerns in medical encounters; however, this has received little attention in physiotherapy. This study explored how physiotherapists solicit and respond to the agenda of concerns that patients with NSCLBP bring to initial encounters. Method: The research setting was primary care. Twenty initial physiotherapy consultations were video-recorded, transcribed and analyzed using conversation analysis, a qualitative observational method. Both verbal and non-verbal features of the interaction were considered. Results: This data highlights a spectrum of communication styles ranging from more physiotherapist-focused, where the physiotherapists did not attend to patients' concerns, to a more patient-focused style, which provided greater opportunities for patients to voice their concerns. On occasions, patients were willing to pursue their own agenda when their concern was initially overlooked. Conclusion: This study provides empirical evidence on communication patterns in physiotherapy practice. A more collaborative style of communication with a shared conversational agenda provided patients with the conversational space to describe their concerns more fully.


Osteopontin plays a unique role in resistance of CD34+/CD123+ human leukemia cell lines KG1a to parthenolide.
To determine if parthenolide (PTL) is cytotoxic for leukemia-like KG1a cells and if it involves in certain molecular-mediated resistance, especially osteopontin (OPN). PTL/daunorubicin (DNR)-treated KG1a cells were examined for viability using MTT and colony-formation assay, and stained for apoptosis using AV/PI. The gene and protein expression were evaluated by qReal-time PCR and Western blotting analysis, respectively. OPN gene was inhibited by OPN siRNA. The cells were stained for various fractions using PE anti-CD34, FITC anti-CD38 and PerCP anti-CD123. Cell viability and proliferation assay exhibited KG1a cells are relatively refractory to used concentrations of PTL. OPN mRNA and protein levels increased in response to PTL. Suppression of OPN with siRNA increased the cytotoxic effects of PTL on KG1a cells. PTL treatment and OPN siRNA suppression in KG1a cells resulted in a decrease of mRNA expression of AKT, mTOR, β-catenin, and Phosphatase and tensin homolog (PTEN). The sub-population cells of CD34+ and CD123+ from KG1a cells are enriched by PTL treatment. Parthenolide in spite of the reduction in gene expression of AKT, mTOR or beta-catenin, stimulates the OPN expression in KG1a cells. The OPN expression pattern in KG1a cells could be compatible with CD34+/CD123+ subtype enrichment by PTL which in turn implies OPN's unique role in resistance of cell populations characterized by CD34+/CD123+ phenotype.


A Conceptual Model of Biopsychosocial Mechanisms of Transition from Acute to Chronic Postsurgical Pain in Children and Adolescents.
Acute and chronic pain are highly prevalent and impactful consequences of surgery across the lifespan, yet a comprehensive conceptual model encompassing biopsychosocial factors underlying acute to chronic pain transition is lacking, particularly in youth. Building on prior chronic postsurgical pain models, we propose a new conceptual model of biopsychosocial mechanisms of transition from acute to chronic postsurgical pain. This review aims to summarize existing research examining key factors underlying acute to chronic postsurgical pain transition in order to guide prevention and intervention efforts aimed at addressing this health issue in children. As pain transitions from acute nociceptive pain to chronic pain, changes in the peripheral and central nervous system contribute to the chronification of pain after surgery. These changes include alterations in sensory pain processing and psychosocial processes (psychological, behavioral, and social components), which promote the development of chronic pain. Patient-related premorbid factors (eg, demographic factors, genetic profile, and medical factors such as premorbid pain) may further modulate these changes. Factors related to acute injury and recovery (eg, surgical and treatment factors), as well as biological response to surgery (eg, epigenetic, inflammatory, and endocrine factors), may also influence this process. Overall, longitudinal studies examining temporal pathways of biopsychosocial processes including both risk and resiliency factors will be essential to identify the mechanisms involved in the transition from acute to chronic pain. Research is also needed to unravel connections between the acute pain experience, opioid exposure, and sensory pain processing during acute to chronic pain transition. Furthermore, future studies should include larger and more diverse samples to more fully explore risk factors in a broader range of pediatric surgeries. The use of conceptual models to guide intervention approaches targeting mechanisms of transition from acute to chronic pain will significantly advance this field and improve outcomes for children and adolescents undergoing surgery.


Length of stay in different drug using states: lifestyles of problem and recreational drug consumers.
The hidden nature of the use of opioids, amphetamines, cocaine, and cannabis (the most commonly used and studied illicit drugs) precludes the accurate estimation of how many people use them and further important aspects of the use. Among the many aspects of drug use, the permanence times in specific drug using states related to the lifestyles of the various sub populations of users are of interest, per se, as essential components of estimates relating prevalence and incidence of drug use and for evaluating policies. Several administrative databases and surveys collected in Italy and in different EU countries concerning different sub-populations of drug users were analysed to estimate the permanence time in a specific drug using state and the stages of a typical drug-using career. Cannabis is mostly the first illegal substance used. The age at onset corresponds to the very beginning of adolescence and various sources indicate that this onset happens earlier for men than for women. For problematic drug users, the switch from cannabis to heavy drugs happens within a rather narrow period of time, within 5-6 years from the first use of cannabis, along with the first episode of drug dealing. The latency period distribution estimated from different data sources is quite stable, with men presenting longer latency periods than women. The comparative analysis of several data sources allowed a more complete overview of the different subpopulations on the drug scene and provides interesting data to understand and to estimate parameters for dynamic models of drug use.


Marine biomonitoring with eDNA: Can metabarcoding of water samples cut it as a tool for surveying benthic communities?
In the marine realm, biomonitoring using environmental DNA (eDNA) of benthic communities requires destructive direct sampling or the setting-up of settlement structures. Comparatively much less effort is required to sample the water column, which can be accessed remotely. In this study we assess the feasibility of obtaining information from the eukaryotic benthic communities by sampling the adjacent water layer. We studied two different rocky-substrate benthic communities with a technique based on quadrat sampling. We also took replicate water samples at four distances (0, 0.5, 1.5, and 20 m) from the benthic habitat. Using broad range primers to amplify a ca. 313 bp fragment of the cytochrome oxidase subunit I gene, we obtained a total of 3,543 molecular operational taxonomic units (MOTUs). The structure obtained in the two environments was markedly different, with Metazoa, Archaeplastida and Stramenopiles being the most diverse groups in benthic samples, and Hacrobia, Metazoa and Alveolata in the water. Only 265 MOTUs (7.5%) were shared between benthos and water samples and, of these, 180 (5.1%) were identified as benthic taxa that left their DNA in the water. Most of them were found immediately adjacent to the benthos, and their number decreased as we moved apart from the benthic habitat. It was concluded that water eDNA, even in the close vicinity of the benthos, was a poor proxy for the analysis of benthic structure, and that direct sampling methods are required for monitoring these complex communities via metabarcoding.


Laparoscopic splenectomy: comparison between anterior and lateral approaches.
Splenectomy, except for a traumatic purpose, is now performed through a laparoscopic approach. There are mainly two ways for laparoscopic total or partial splenectomies. For the classic anterior dissection of the splenic vessels, patient is placed in supine position and five ports are required to elevate the spleen and proceed to vessel divisions. With a lateral approach of the pedicle, patient is placed in lateral decubitus position and three ports are sufficient, because gravity help to provide traction on the splenic ligaments and to present hilar vessels and pancreas tail. The aim of our study was to compare surgical complications of those two approaches of laparoscopic splenectomy in children. We reviewed 84 medical records of patient operated on for hematological disease between January 1993 and December 2009. There were 47 anterior and 37 lateral approaches. Sex, disease, median age, operative time, blood lost or hospital stay, and associated laparotomy were not different between the two groups. Operative complications included hemorrhage (5), bowel injury (1), diaphragmatic wound (1), pancreas tail section (1), and parietal hematoma (1) in the anterior group (9 cases) versus 1 hemorrhage in the lateral group (P<.02). There were five laparotomies owing to surgical complications in the anterior group, and none in the lateral group. Splenectomy through laparoscopic approach is an effective technique. Lateral dissection of the vessels provides less operative complications in children.


Ultrasonographic image of fatty infiltration of the liver correlates with selected biochemical parameters and back fat thickness of periparturient Holstein-Friesian cows.
During the transition period, the cow's body activates adaptive mechanisms aimed at adjusting to the changing demand for energy and nutrients, which are necessary for the growing fetus and the subsequent start of milk production. This time is also associated with an increased risk of metabolic diseases and reproductive disorders. Our study aimed to identify prepartum and postpartum biochemical markers and weight loss patterns that could differentiate cows that would exhibit ultrasonographic signs of liver fatty infiltration during the latter half of the transition period. The study was performed in a single herd of Holstein-Friesian cows and the animals were divided into two groups: CON (n=13) - cows without ultrasonographic signs of fatty liver, and FL (n=16) - cows with ultrasonographic signs of fatty liver. Backfat thickness and specific biochemical parameters were measured weekly from one week before parturition to 9 weeks postpartum. Our study highlights the importance of using a combination of monitoring methods to assess the metabolic status of transition dairy cattle. The results showed that ultrasound measurements of backfat thickness, blood NEFA levels, glucose concentration, and AST activity were all different (p<0.05) between the control and FL groups, indicating the usefulness of these parameters in monitoring the health status of transition cows. Additionally, the results suggest that high prepartum glucose levels (4.99 mmol/l) could serve as a potential marker for future FL, while the elevated NEFA levels (0.51 mmol/l) and decreased AST activity (80.56 u/l) in FL animals indicate their potential as indicators of lipid mobilization and liver structural damage, respectively.


Low Yield of Cerebral Angiography in Adequately Occluded Aneurysms After Flow Diversion.
Flow diversion has emerged as a highly effective treatment for intracranial aneurysms. To assess the yield of further angiographic follow-up in aneurysms that have achieved adequate occlusion after treatment with the Pipeline Embolization Device (PED; Medtronic Inc, Dublin, Ireland). This is a single-institution, retrospective study. Inclusion criteria were as follows: (1) patients with 1 or more aneurysms treated with PED, (2) available short-term (<12 mo) follow-up digital subtraction angiography (DSA), (3) complete (100%) or near-complete (>95%) occlusion on short-term follow-up DSA, and (4) available further angiographic follow-up (DSA, Magnetic Resonance Angiography (MRA), or Computed Tomography Angiography (CTA)). A total of 146 patients were identified. Aneurysm size was 8.4 ± 5.1 mm on average. Mean angiographic follow-up time was 29.7 ± 12.2 mo. On short-term follow-up DSA images, 132 (90.4%) had complete aneurysm occlusion and 14 (9.6%) had near-complete occlusion. Four patients (3%) had further DSA follow-up alone, 30 patients (21%) had further DSA and MRA/CTA follow-up, and 112 patients (76%) had further MRA/CTA follow-up alone. On further angiographic follow-up (DSA, MRA, and/or CTA), no patient had a decrease in the degree of aneurysm occlusion (recurrence) or required retreatment. Of the 14 patients with near-complete occlusion on initial DSA images, 7 patients (50%) progressed to complete aneurysm occlusion on further angiographic follow-up. This study did not find any diagnostic yield in repeating cerebral angiography in adequately occluded aneurysms with the PED. We do not recommend repeat angiographic follow-up once aneurysms have achieved complete occlusion with the PED unless clinically warranted.


Fluorescence-based multi-parameter approach to characterize dynamics of organic carbon, faecal bacteria and particles at alpine karst springs.
Karst springs, especially in alpine regions, are important for drinking water supply but also vulnerable to contamination, especially after rainfall events. This high variability of water quality requires rapid quantification of contamination parameters. Here, we used a fluorescence-based multi-parameter approach to characterize the dynamics of organic carbon, faecal bacteria, and particles at three alpine karst springs. We used excitation emission matrices (EEMs) to identify fluorescent dissolved organic material (FDOM). At the first system, peak A fluorescence and total organic carbon (TOC) were strongly correlated (Spearman's rs of 0.949), indicating that a large part of the organic matter is related to humic-like substances. Protein-like fluorescence and cultivation-based determination of coliform bacteria also had a significant correlation with rs=0.734, indicating that protein-like fluorescence is directly related to faecal pollution. At the second system, which has two spring outlets, the absolute values of all measured water-quality parameters were lower; there was a significant correlation between TOC and humic-like fluorescence (rs=0.588-0.689) but coliform bacteria and protein-like fluorescence at these two springs were not correlated. Additionally, there was a strong correlation (rs=0.571-0.647) between small particle fractions (1.0 and 2.0μm), a secondary turbidity peak and bacteria. At one of these springs, discharge was constant despite the reaction of all other parameters to the rainfall event. Our results demonstrated that i) all three springs showed fast and marked responses of all investigated water-quality parameters after rain events; ii) a constant discharge does not necessarily mean constant water quality; iii) at high contamination levels, protein-like fluorescence is a good indicator of bacterial contamination, while at low contamination levels no correlation between protein-like fluorescence and bacterial values was detected; and iv) a combination of fluorescence measurements and particle-size analysis is a promising approach for a rapid assessment of organic contamination, especially relative to time-consuming conventional bacterial determination methods.


[Role of the basal ganglia in the occurrence of paradoxical sleep dreams (hypothetical mechanism)].
A hypothetical mechanism of the basal ganglia involvement in the occurrence of paradoxical sleep dreams and rapid eye movements is proposed. According to this mechanism, paradoxical sleep is provided by facilitation of activation of cholinergic neurons in the pedunculopontine nucleus as a result of suppression of their inhibition from the output basal ganglia nuclei. This disinhibition is promoted by activation of dopaminergic cells by pedunculopontine neurons, subsequent rise in dopamine concentration in the input basal ganglia structure. striatum, and modulation of the efficacy of cortico-striatal inputs. In the absence of signals from retina, a disinhibition of neurons in the pedunculopontine nucleus and superior colliculus allows them to excite neurons in the lateral geniculate body and other thalamic nuclei projecting to the primary and higher visual cortical areas, prefrontal cortex and back into the striatum. Dreams as visual images and "motor hallucinations" are the result of an increase in activity of definitely selected groups of thalamic and neocortical neurons. This selection is caused by modifiable action of dopamine on long-term changes in the efficacy of synaptic transmission during circulation of signals in closed interconnected loops, each of which includes one of the visual cortical areas (motor cortex), one of the thalamic nuclei, limbic and one of the visual areas (motor area) of the basal ganglia. pedunculopontine nucleus, and superior colliculus. Simultaneous modification and modulation of synapses in diverse units of neuronal loops is provided by PGO waves. Disinhibition of superioir colliculus neurons and their excitation by pedunculopontine nucleus lead to an appearance of rapid eye movements during paradoxical sleep.


Discs Large Homolog 1 Splice Variants Regulate p38-Dependent and -Independent Effector Functions in CD8+ T Cells.
Functionally diverse CD8+ T cells develop in response to antigenic stimulation with differing capacities to couple TCR engagement to downstream signals and functions. However, mechanisms of diversifying TCR signaling are largely uncharacterized. Here we identified two alternative splice variants of scaffold protein Dlg1, Dlg1AB and Dlg1B, that diversify signaling to regulate p38-dependent and -independent effector functions in CD8+ T cells. Dlg1AB, but not Dlg1B associated with Lck, coupling TCR stimulation to p38 activation and proinflammatory cytokine production. Conversely, both Dlg1AB and Dlg1B mediated p38-independent degranulation. Degranulation depended on a Dlg1 fragment containing an intact Dlg1SH3-domain and required the SH3-ligand WASp. Further, Dlg1 controlled WASp activation by promoting TCR-triggered conformational opening of WASp. Collectively, our data support a model where Dlg1 regulates p38-dependent proinflammatory cytokine production and p38-independent cytotoxic granule release through the utilization of alternative splice variants, providing a mechanism whereby TCR engagement couples downstream signals to unique effector functions in CD8+ T cells.


Acute vasodilator effect of fasudil, a Rho-kinase inhibitor, in monocrotaline-induced pulmonary hypertension in rats.
Pulmonary arterial hypertension is a progressive and fatal disease for which Rho-kinase may be substantially involved. In this study, we examined the acute vasodilator effects of fasudil, a Rho-kinase inhibitor, in monocrotaline (MCT)-induced pulmonary hypertension (PH) in rats. Three weeks after a single subcutaneous injection of MCT (60 mg/kg), hemodynamic variables were measured under conscious and free-moving conditions before and after oral administration of fasudil. MCT caused a significant elevation of mean pulmonary arterial pressure (mPAP). Although a low dose of fasudil (3 mg/kg) had no effect on mPAP, a middle dose (10 mg/kg) caused a significant reduction in mPAP without change in mean systemic arterial pressure (mSAP), and a high dose (30 mg/kg) significantly reduced both mPAP and mSAP. Rho-kinase activity was significantly increased by MCT injection in pulmonary arteries but not in the aorta. Fasudil (10 mg/kg) inhibited only the Rho-kinase activity in pulmonary arteries without any effect in the aorta. Plasma concentration of hydroxyfasudil, a metabolite of fasudil, was within its clinical range in humans. These results demonstrate that fasudil exerts effective and selective vasodilatation of pulmonary arteries in rats with MCT-induced PH at a given dose, suggesting its usefulness for the treatment of the fatal disorder.


Friedreich Ataxia: Developmental Failure of the Dorsal Root Entry Zone.
Dorsal root ganglia, dorsal roots (DR), and dorsal root entry zones (DREZ) are vulnerable to frataxin deficiency in Friedreich ataxia (FA). A previously unrecognized abnormality is the intrusion of astroglial tissue into DR. Segments of formalin-fixed upper lumbar spinal cord of 13 homozygous and 2 compound heterozygous FA patients were sectioned longitudinally to represent DREZ and stained for glial fibrillary acidic protein (GFAP), S100, vimentin, the central nervous system (CNS)-specific myelin protein proteolipid protein, the peripheral nervous system (PNS) myelin proteins PMP-22 and P0, and the Schwann cell proteins laminin, alpha-dystroglycan, and periaxin. Normal DREZ showed short, sharply demarcated, dome-like extensions of CNS tissue into DR. The Schwann cell-related proteins formed tight caps around these domes. In FA, GFAP-, S100-, and vimentin-reactive CNS tissue extended across DREZ and into DR over much longer distances by breaching the CNS-PNS barrier. The transition between PNS and CNS myelin proteins was disorganized. During development, neural-crest derived boundary cap cells provide guidance to dorsal root ganglia axons growing into the dorsal spinal cord and at the same time block the inappropriate intrusion of CNS glia into DR. It is likely that frataxin is required during a critical period of permissive (axons) and nonpermissive (astroglia) border-control.


Association of atrial fibrillation with mortality and disability after ischemic stroke.
We determined whether patient characteristics (age, sex, comorbidities), stroke severity, and quality of care explained a proportion of the association between atrial fibrillation (AF) and increased disability and mortality in patients with acute ischemic stroke. The study included a prospective cohort of consecutive patients admitted with acute ischemic stroke included in the Registry of the Canadian Stroke Network (July 1, 2003 to March 31, 2008). Multivariable logistic regression analyses were used to determine the magnitude of association between AF and modified Rankin score 4-5 at discharge, 30-day mortality, and 1-year mortality. There were 10,528 patients admitted with acute ischemic stroke. AF was associated with an increased risk of severe disability and mortality, but the magnitude of association was substantially attenuated in the full multivariable models: modified Rankin score 4-5 at discharge (univariate odds ratio [OR] 1.74, 95% confidence interval [CI] 1.57-1.93; multivariable OR 1.19, 95% CI 1.03-1.36), 30-day mortality (univariate OR 2.52, 95% CI 2.25-2.84; multivariable OR 1.36, 95% CI 1.17-1.58), and 1-year mortality (univariate OR 2.41, 95% CI 2.19-2.66; multivariable OR 1.25, 95% CI 1.10-1.42). Older age and increased stroke severity explained most of the association between AF and poor stroke outcomes. We found no association between AF and poor stroke outcomes in patients receiving therapeutic preadmission oral anticoagulant therapy. Older age and increased stroke severity explain most of the association between AF and poorer outcomes after acute ischemic stroke. Nonuse of oral anticoagulant therapy represents the most important modifiable care gap to mitigate the association between AF and poor outcomes after ischemic stroke.


Influence of sex on sleep regulatory mechanisms.
The ability of biological sex and sex-driven characteristics to alter sleep states may contribute to gender disparities in sleep disorders. Sex influences sleep-wake amount, the daily timing of the sleep-wake cycle, and the ability to restore sleep after extended wakefulness. Several lines of evidence suggest that in mammals, reproductive hormones are responsible for the effects of sex on sleep and may have organizational and activational influences on sleep regulatory mechanisms. In humans, exogenously administered estrogens and progestins generally enhance sleep amount and continuity, whereas androgens appear to have a positive impact on rapid eye movement (REM) sleep but disrupt sleep consolidation. In rodent studies, however, female reproductive hormones appear to enhance wakefulness, and male gonadal hormones reinforce sleep. Rodent studies have also revealed that neonatal exposure to reproductive hormones organizes adult sleep-wake architecture. This paper reviews how sex and reproductive hormones interact with circadian and homeostatic sleep regulatory mechanisms in humans and animal models. We examine the organizational and activational nature of these interactions and also review how these interactions change with advancing age. Finally, we discuss the potential for genetic sex to influence sleep states. It is our hope that a better understanding of the mechanisms through which sex influences sleep-wake states will lead to improvements in the design of studies that examine gender disparities in sleep-wake disorders.


Functional characterization of sodium-pumping rhodopsins with different pumping properties.
Sodium pumping rhodopsins (NaRs) are a unique member of the microbial-type I rhodopsin family which actively transport Na+ and H+ depending on ionic condition. In this study, we surveyed 12 different NaRs from various sources of eubacteria for their electrophysiological as well as spectroscopic properties. In mammalian cells several of these NaRs exhibited a Na+ based pump photocurrent and four interesting candidates were chosen for further characterization. Voltage dependent photocurrent amplitudes revealed a membrane potential-sensitive turnover rate, indicating the presence of an electrically-charged intermediate(s) in the photocycle reaction. The NaR from Salinarimonas rosea DSM21201 exhibited a red-shifted absorption spectrum, and slower kinetics compared to the first described sodium pump, KR2. Although the ratio of Na+ to H+ ion transport varied among the NaRs we tested, the NaRs from Flagellimonas sp_DIK and Nonlabens sp_YIK_SED-11 showed significantly higher Na+ selectivity when compared to KR2. All four further investigated NaRs showed a functional expression in dissociated hippocampal neuron culture and hyperpolarizing activity upon light-stimulation. Additionally, all four NaRs allowed optical inhibition of electrically-evoked neuronal spiking. Although efficiency of silencing was 3-5 times lower than silencing with the enhanced version of the proton pump AR3 from Halorubrum sodomense, our data outlines a new approach for hyperpolarization of excitable cells without affecting the intracellular and extracellular proton environment.


Effect of maturity stage and storage on flavor compounds and sensory description of berrycactus (Myrtillocactus geometrizans).
Descriptive sensory analysis combined with a chemical analysis, provided insight regarding sensory significance for a better understanding of berrycactus' flavor. This study was based on the volatile components derived from the gas chromatography-olfactometry (GC-O) analysis. Four different ripening stages were used for the experiments: unripe, red, transition, and over-ripe. Besides red and transition stages were stored in sealed plastic bags for 7 d at 5 °C as an additional treatment in order to establish comparisons with fresh samples. GC-O analysis was performed following OSME (time-intensity) method. Moreover, chemical characterization was achieved through GC-MS analysis, each compound was tentatively identified by comparing its mass spectra with the spectra from the library NIST02, by its aroma notes and Kovats Index. The aroma descriptors found were: caramel, fruity, acetic acid, fresh, citrus, floral, and phenolics. Nine volatile compounds are among the most important in flavor of berrycactus: furfural, 5-methyl-2-furancarboxaldehyde, 2(5H)-furanone, 5-acetoxymethyl-2-furaldehyde, 2-cyclohexen-1-ol, octanoic acid ethyl ester, decanoic acid ethyl ester, octanoic acid, and phenylethyl alcohol. To better reflect the real responses of these compounds, the regression analysis of concentrations to ripening stages were performed. The 2,3-dihydro-3,5-dihydroxy-6-methyl-(4H)-pyran-4-one was found in berrycactus, it has been reported in chemical profile of other berries as a product of heat-induced reactions, however, this compound has no aroma activity. The data suggest that both furanones and esters contribute to the aroma of berrycactus. On the other hand, solid phase microextraction was found to be more useful in detecting esters and acids whereas solvent extraction was more effective in detecting furanones and ketones. This information could contribute to setting up favorable processing conditions; in order to retain the best sensory characteristics of berrycactus along with functional properties that would enhance commercialization and exploitation of this fruit and to promote berrycactus cultivation programs.