[The effect of FK 506, an immunosuppressant, on cerebral infarction volume in focal cerebral ischemia in rats].
The effect of FK 506, an immunosuppressant which is widely used in the transplantation of liver, kidney and bone marrow, on cerebral infarction volume in transient focal ischemia was investigated. Focal cerebral ischemia was produced by 2 h occlusion of the left middle cerebral artery in rats. FK 506 (0.3 mg/kg body weight) and vehicle were administered intravenously immediately after induction of cerebral ischemia. After 24 h reperfusion the rats were sacrificed and brain infarct volume and edema volume were determined. The FK 506 treatment group showed a significantly reduced infarct volume in the cerebral cortex when it was compared with the vehicle treatment group, but infarct volume was not significantly reduced in the striatum which was the ischemic core in this focal ischemia model. However, FK 506 did not reduce the edema volume significantly. These results suggest that the reduction of infarct volume is a result of the neuroprotective effect of FK 506. This immunosuppressant may be useful in the treatment of cerebral infarction.


[Stiff person syndrome: still a difficult and overlooked diagnosis].
We present a case of stiff person syndrome (SPS) with a description of the diagnostic challenges and a discussion of the implication of novel antibodies. SPS is a rare neurological disorder presenting with severe rigidity, muscle spasms and impaired gait function, and diagnosis is often delayed due to low awareness. SPS is classically associated with antibodies directed against glutamic acid decarboxylase or amphiphysin. However, recent advances have contributed to our understanding because of detection of several new antibodies, which cause SPS, mostly with additional neurological/systemic manifestations.


The "Twitching Technique": A New Space-Irrespective Laparoscopic Ligation Technique Using a JAiMY Needle Holder.
Background: Laparoscopic intracorporeal sutures and knots require advanced techniques in children. The JAiMY® needle holder (Endocontrol Company, Grenoble, France), a flexible jaw with both clockwise and anticlockwise revolving functions, enables the placement of sutures in very small working spaces. Using this tool, a novel new ligation method, which we named "twitching technique," can be performed. Methods: To perform the "twitching technique," first grasp the long tail of the thread with the JAiMY and the forceps on the needle side. Then, rotate the tip of the needle holder halfway in either direction and bring the forceps closer to the needle holder to make a ring using the thread. Grasp the thread at the crossing point with the forceps, and then catch the short tail through the ring with the needle holder. Results (Case Presentation): A 1-year-old boy presented with vomiting because of esophageal hiatal hernia. He was being treated for single ventricle and asplenia at our hospital. Laparoscopic esophageal hiatus plication was performed. Although the working space was extremely limited, the JAiMY and the "twitching technique" enabled steady suture placement and ligation. Conclusions: The "twitching technique" is an easy, steady, and safe method for performing ligation in difficult situations and may facilitate the performance of many laparoscopic surgeries in pediatric fields.


Classification of changes occurring in lung patient during radiotherapy using relative γ analysis and hidden Markov models.
To present a new automated patient classification method based on relative gamma analysis and hidden Markov models (HMM) to identify patients undergoing important anatomical changes during radiation therapy. Daily EPID images of every treatment field were acquired for 52 patients treated for lung cancer. In addition, CBCT were acquired on a regular basis. Gamma analysis was performed relative to the first fraction given that no significant anatomical change was observed on the CBCT of the first fraction compared to the planning CT. Several parameters were extracted from the gamma analysis (e.g., average gamma value, standard deviation, percent above 1). These parameters formed patient-specific time series. Data from the first 24 patients were used as a training set for the HMM. The trained HMM was then applied to the remaining 28 patients and compared to manual clinical evaluation and fixed thresholds. A three-category system was used for patient classification ranging from minor deviations (category 1) to severe deviations (category 3) from the treatment plan. Patient classified using the HMM lead to the same result as the classification made by a human expert 83% of the time. The HMM overestimate the category 10% of the time and underestimate 7% of the time. Both methods never disagree by more than one category. In addition, the information provided by the HMM is richer than the simple threshold-based approach. HMM provides information on the likelihood that a patient will improve or deteriorate as well as the expected time the patient will remain in that state. We showed a method to classify patients during the course of radiotherapy based on relative changes in EPID images and a hidden Markov model. Information obtained through this automated classification can complement the clinical information collected during treatment and help identify patients in need of a plan adaptation.


Interconversion of components of the bacterial proton motive force by electrogenic potassium transport.
The influence of K+ ions on the components of the transmembrane proton motive force (delta mu H+) in intact bacteria was investigated. In K+-depleted cells of the glycolytic bacterium STreptococcus faecalis the addition of K+ ions caused a depolarization of the membrane by about 60 mV. However, since the depolarization was compensated for by an increase in the transmembrane pH gradient (delta pH), the total proton motive force remained almost constant at about 120 mV. Half-maximal changes in the potential were observed at K+ concentrations at which the cells accumulated K+ ions extensively. In EDTA-treated, K+-depleted cells of Escherichia coli K-12, the addition of K+ ions to the medium caused similar, although smaller changes in the components of delta mu H+. Experiments with various E. coli K-12 K+ transport mutants showed that for the observed potential changes the cells required either a functional TrkA or Kdp K+ transport system. These data are interpreted to mean that the inward movement of K+ ions via each of these bacterial transport systems is electrogenic. Consequently, it leads to a depolarization of the membrane, which in its turn allows the cell to pump more protons into the medium.


Chronological changes in acid phosphatase activity within neurons and perineuronal satellite cells of the inferior vagal ganglion of the cat induced by vagotomy.
The hexazonium pararosaniline method was employed to describe the distribution of acid phosphatase activity, chronologically, within neurons and their investing satellite cells of the inferior vagal ganglion of the cat after vagotomy. In control ganglia, acid phosphatase activity was invariably confined to the cytoplasm of neurons and satellite cells. Reaction product was visible as distinct granules within neuronal perikarya. The cytoplasm of perineuronal satellite cells also contained reaction product but, in most instances, activity was weak and granules were difficult to distinguish. No reaction product was observed in myelin or axonal processes; nuclear staining was absent. Acid phosphatase activity was increased in ganglionic neurons as early as 24 hours after vagotomy. Increased activity in perineuronal satellite cells was not evident until 3 days post-operatively. By 15 days, activity was ubiquitously increased in the cytoplasm of both neurons and satellite cells. Evidence suggesting neuronophagia was also apparent. Between 30 and 60 days post-operatively acid phosphatase activity gradually decreased in both neurons and satellite cells until a picture comparable with that seen in control tissue sections was visible. The functional significance of these changes in acid phosphatase activity within an altered metabolic environment induced by vagotomy is discussed.


Current management of esophageal perforation: 20 years experience.
Esophageal perforations are surgical emergencies associated with high morbidity and mortality rates. No single strategy has been sufficient to deal with the majority of situations. We aim to postulate a therapeutic algorithm for this complication based on 20 years of experience and also on data from published literature. We performed a retrospective clinical review of 44 patients treated for esophageal perforation at our hospital between January 1989 and May 2008. We reviewed the characteristics of these patients, including age, gender, accompanying diseases, etiology of perforation, diagnosis, location, time interval between perforation and diagnosis, treatment of the perforation, morbidity, hospital mortality, and duration of hospitalization. Perforation occurred in the cervical esophagus in 14 patients (31.8%), thoracic esophagus in 18 patients (40.9%), and abdominal esophagus in 12 patients (27.3%). Management of the esophageal perforation included primary closure in 23 patients (52.3%), resection in 7 patients (15.9%), and nonsurgical therapy in 14 patients (31.8%). In the surgically treated group, the mortality rate was 3 of 30 patients (10%), and 2 of 14 patients (14.3%) in the conservatively managed group. Four of the 14 nonsurgical patients were inserted with covered self-expandable stents. The specific treatment of an esophageal perforation should be selected according to each individual patient. To date, the most effective treatment would appear to be operative management. With improvements in endoscopic procedures, the morbidity and mortality rates of esophageal perforations are significantly decreased. We suggest that minimally invasive techniques for the repair of esophageal perforations will be very important in the future treatment of this condition.


BPR1J-097, a novel FLT3 kinase inhibitor, exerts potent inhibitory activity against AML.
Activating mutations of Fms-like tyrosine kinase 3 (FLT3) constitute a major driver in the pathogenesis of acute myeloid leukaemia (AML). Hence, pharmacological inhibitors of FLT3 are of therapeutic interest for AML. The effects of inhibition of FLT3 activity by a novel potent FLT3 inhibitor, BPR1J-097, were investigated using in vitro and in vivo assays. The 50% inhibitory concentration (IC(50)) of BPR1J-097 required to inhibit FLT3 kinase activity ranged from 1 to 10 nM, and the 50% growth inhibition concentrations (GC(50)s) were 21±7 and 46±14 nM for MOLM-13 and MV4-11 cells, respectively. BPR1J-097 inhibited FLT3/signal transducer and activator of transcription 5 phosphorylation and triggered apoptosis in FLT3-driven AML cells. BPR1J-097 also showed favourable pharmacokinetic property and pronounced dose-dependent tumour growth inhibition and regression in FLT3-driven AML murine xenograft models. These results indicate that BPR1J-097 is a novel small molecule FLT-3 inhibitor with promising in vivo anti-tumour activities and suggest that BPR1J-097 may be further developed in preclinical and clinical studies as therapeutics in AML treatments.


Association of CYP2A6 activity with lung cancer incidence in smokers: The multiethnic cohort study.
While smoking is the primary cause of lung cancer, only 11-24% of smokers develop the malignancy over their lifetime. The primary addictive agent in tobacco smoke is nicotine and variation in nicotine metabolism may influence the smoking levels of an individual. Therefore, inter-individual variation in lung cancer risk among smokers may be due in part to differences in the activity of enzymes involved in nicotine metabolism. In most smokers, cytochrome P450 2A6 (CYP2A6)-catalyzed C-oxidation accounts for >75% of nicotine metabolism, and the activity of this enzyme has been shown to correlate with the amount of nicotine and carcinogens drawn from cigarettes. We prospectively evaluated the association of urinary biomarkers of nicotine uptake (total nicotine equivalents [TNE]) and CYP2A6 activity (ratio of urinary total trans-3'-hydroxycotinine to cotinine) with lung cancer risk among 2,309 Multiethnic Cohort Study participants who were current smokers at time of urine collection; 92 cases were diagnosed during a mean follow-up of 9.5 years. We found that higher CYP2A6 activity and TNE was associated with increased lung cancer risk after adjusting for age, sex, race/ethnicity, body mass index, smoking duration, and urinary creatinine (p's = 0.002). The association for CYP2A6 activity remained even after adjusting for self-reported cigarettes per day (CPD) (Hazard Ratio [HR] per unit increase in log-CYP2A6 activity = 1.52; p = 0.005) and after adjusting for TNE (HR = 1.46; p = 0.01). In contrast, the association between TNE and lung cancer risk was of borderline statistical significance when adjusted for CPD (HR = 1.53; p = 0.06) and not statistically significant when further adjusted for CYP2A6 activity (HR = 1.30; p = 0.22). These findings suggest that CYP2A6 activity provides information on lung cancer risk that is not captured by smoking history or a (short-term) biomarker of dose. CYP2A6 activity should be further studied as a risk biomarker for smoking-related lung cancer.


Unpredictable chronic stress decreases inhibitory avoidance learning in Tuebingen long-fin zebrafish: stronger effects in the resting phase than in the active phase.
Zebrafish (Danio rerio Hamilton) are increasingly used as a model to study the effects of chronic stress on brain and behaviour. In rodents, unpredictable chronic stress (UCS) has a stronger effect on physiology and behaviour during the active phase than during the resting phase. Here, we applied UCS during the daytime (active phase) for 7 and 14 days or during the night-time (resting phase) for 7 nights in an in-house-reared Tuebingen long-fin (TLF) zebrafish strain. Following UCS, inhibitory avoidance learning was assessed using a 3 day protocol where fish learn to avoid swimming from a white to a black compartment where they will receive a 3 V shock. Latencies of entering the black compartment were recorded before training (day 1; first shock) and after training on day 2 (second shock) and day 3 (no shock, tissue sampling). Fish whole-body cortisol content and expression levels of genes related to stress, fear and anxiety in the telencephalon were quantified. Following 14 days of UCS during the day, inhibitory avoidance learning decreased (lower latencies on days 2 and 3); minor effects were found following 7 days of UCS. Following 7 nights of UCS, inhibitory avoidance learning decreased (lower latency on day 3). Whole-body cortisol levels showed a steady increase compared with controls (100%) from 7 days of UCS (139%), to 14 days of UCS (174%) to 7 nights of UCS (231%), suggestive of an increasing stress load. Only in the 7 nights of UCS group did expression levels of corticoid receptor genes (mr, grα, grβ) and of bdnf increase. These changes are discussed as adaptive mechanisms to maintain neuronal integrity and prevent overload, and as being indicative of a state of high stress load. Overall, our data suggest that stressors during the resting phase have a stronger impact than during the active phase. Our data warrant further studies on the effect of UCS on stress axis-related genes, especially grβ; in mammals this receptor has been implicated in glucocorticoid resistance and depression.


Antihypertensive effect of all-cis-5,8,11,14,17-icosapentaenoate of aged rats is associated with an increase in the release of ATP from the caudal artery.
Fish oils have been shown to lower blood pressure in hypertensive subjects. All-cis-5,8,11,14,17-icosapentaenoate (EPA), one of the omega-3 polyunsaturated fatty acids, is known to be one of the major active components in fish oil that has beneficial effects on the cardiovascular system. However, little is known about the antihypertensive effect of EPA alone on blood pressure. In the present study, we have determined the spontaneous and noradrenaline-evoked release of ATP, ADP, AMP, and adenosine from caudal arteries of aged (100 weeks old) Wistar rats which were fed a standard diet or a high cholesterol diet, treated with EPA. Dietary EPA administration increased plasma and caudal arterial EPA concentrations and repressed increases in blood pressure with advancing age in both aged rats with and without hypercholesterolemia. In addition, noradrenaline (1 micromol/l) evoked a significantly greater release of purines from the caudal arteries of EPA-administered aged rats compared to both sets of control rats. Regression analysis revealed a significant relationship between the total amount of purines released from the artery and blood pressure. These results suggest that administration of EPA to aged rats increases the release of ATP from the vascular endothelial cells, leading to repression of the blood pressure rise seen with advancing age.


Regulation of GluA1 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor function by protein kinase C at serine-818 and threonine-840.
Three residues within the AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor subunit GluA1 C terminus (Ser818, Ser831, Thr840) can be phosphorylated by Ca(2+)/phospholipid-dependent protein kinase (PKC). Here, we show that PKC phosphorylation of GluA1 Ser818 or Thr840 enhances the weighted mean channel conductance without altering the response time course or agonist potency. These data support the idea that these residues constitute a hyper-regulatory domain for the AMPA receptor. Introduction of phosphomimetic mutations increases conductance only at these three sites within the proximal C terminus, consistent with a structural model with a flexible linker connecting the distal C-terminal domain to the more proximal domain containing a helix bracketed by Ser831 and Thr840. NMR spectra support this model and raise the possibility that phosphorylation can alter the configuration of this domain. Our findings provide insight into the structure and function of the C-terminal domain of GluA1, which controls AMPA receptor function and trafficking during synaptic plasticity in the central nervous system.


Expression patterns of mouse and human CYP orthologs (families 1-4) during development and in different adult tissues.
The present study compared the relative expression pattern of 10 orthologous CYP forms from families 1-4 in cDNA panels of human and mouse fetal and adult tissues. Except for CYP1A2, all of these CYPs exhibited specific patterns of expression during mouse ontogeny, suggesting possible involvement in development. Cyp1a1 and Cyp2r1 were the only two of the orthologs to be expressed only in the E7 mouse; Cyp2s1 was expressed in all stages, including E7, while Cyp2e1 appeared only at E17. Highest expression of the individual CYPs in the different late term human fetal tissues was: thymus, CYP1B1 and CYP2U1; liver, CYP2E1; brain, CYP2R1, CYP1A1 and CYP4X1; and lung, CYP4B1 and CYP2W1. In general, the level of individual human CYP transcripts was lower in the fetal than the corresponding adult tissues. The pattern of expression in adult mouse and human tissues was fairly similar for CYP1A1, CYP1A2, CYP1B1, CYP2S1, and CYP2U1 orthologs.


Different sets of traits determine transition of alien species along the invasion continuum.
Invasive alien species are currently considered as one of the dominant drivers of global environmental change. Till now, the majority of studies have focused on single or a few traits of alien species that facilitate their invasion. Also inclusion of all the traits which determine the transition of aliens along the different stages of invasion continuum (casual, naturalised and invasive) has remained largely overlooked. In this study, we collected a comprehensive trait dataset on 144 alien plant species of Kashmir Himalaya - a global biodiversity hotspot region. To test which traits of alien species, individually or in combination along with anthropogenic factors, determine their transition along the invasion continuum, we employed chi-square tests, boosted regression trees and phylogenetic methods. We found the perennial lifespan, longer residence time, greater number of introduced regions, and better seed dispersal mechanism were critical in determining the transition from casual to naturalised. The herbaceous growthform (therophytes), annual lifespan, achene fruit, longer residence time and broader introduced range were the species' traits determining transition from naturalised to invasive. Aliens introduced as ornamentals have more propensity to become naturalised; whereas aliens introduced unintentionally show overrepresentation at the invasive stage. Phylogeny alone showed mixed results indicating both clustering and dispersion; however in combination with other traits, it plays a significant role in determining the stage of invasion. Overall, our results disentangle the individual and interactive roles of multiple traits that determine the transition of alien species' along the invasion continuum. Further, we foresee the potential applicability of our findings in designing robust invasion risk analysis protocols and stage-specific invasion management strategies in this Himalayan region, with learnings for elsewhere in the world.


The impact of growing-season length variability on carbon assimilation and evapotranspiration over 88 years in the eastern US deciduous forest
Recent research suggests that increases in growing-season length (GSL) in mid-northern latitudes may be partially responsible for increased forest growth and carbon sequestration. We used the BIOME-BGC ecosystem model to investigate the impacts of including a dynamically regulated GSL on simulated carbon and water balance over a historical 88-year record (1900-1987) for 12 sites in the eastern USA deciduous broadleaf forest. For individual sites, the predicted GSL regularly varied by more than 15 days. When grouped into three climatic zones, GSL variability was still large and rapid. There is a recent trend in colder, northern sites toward a longer GSL, but not in moderate and warm climates. The results show that, for all sites, prediction of a long GSL versus using the mean GSL increased net ecosystem production (NEP), gross primary production (GPP), and evapotranspiration (ET); conversely a short GSL is predicted to decrease these parameters. On an absolute basis, differences in GPP between the dynamic and mean GSL simulations were larger than the differences in NEP. As a percentage difference, though, NEP was much more sensitive to changes in GSL than were either GPP or ET. On average, a 1-day change in GSL changed NEP by 1.6%, GPP by 0.5%, and ET by 0.2%. Predictions of NEP and GPP in cold climates were more sensitive to changes in GSL than were predictions in warm climates. ET was not similarly sensitive. First, our results strongly agree with field measurements showing a high correlation between NEP and dates of spring growth, and second they suggest that persistent increases in GSL may lead to long-term increases in carbon storage.


The Core and Holoenzyme Forms of RNA Polymerase from Mycobacterium smegmatis.
Bacterial RNA polymerase (RNAP) is essential for gene expression and as such is a valid drug target. Hence, it is imperative to know its structure and dynamics. Here, we present two as-yet-unreported forms of Mycobacterium smegmatis RNAP: core and holoenzyme containing σA but no other factors. Each form was detected by cryo-electron microscopy in two major conformations. Comparisons of these structures with known structures of other RNAPs reveal a high degree of conformational flexibility of the mycobacterial enzyme and confirm that region 1.1 of σA is directed into the primary channel of RNAP. Taken together, we describe the conformational changes of unrestrained mycobacterial RNAP.IMPORTANCE We describe here three-dimensional structures of core and holoenzyme forms of mycobacterial RNA polymerase (RNAP) solved by cryo-electron microscopy. These structures fill the thus-far-empty spots in the gallery of the pivotal forms of mycobacterial RNAP and illuminate the extent of conformational dynamics of this enzyme. The presented findings may facilitate future designs of antimycobacterial drugs targeting RNAP.


Neural mechanism underlying the beneficial effect of Theory of Mind psychotherapy on early-onset schizophrenia: a randomized controlled trial.
Psychosocial interventions have emerged as an important component of a comprehensive therapeutic approach in early-onset schizophrenia, typically representing a more severe form of the disorder. Despite the feasibility and efficacy of Theory of Mind (ToM) psychotherapy for schizophrenia, relatively little is known regarding the neural mechanism underlying its effect on early-onset schizophrenia. We performed a randomized, active controlled trial in patients with early-onset schizophrenia, who were randomly allocated into either an intervention (ToM psychotherapy) or an active control (health education) group. Diffusion tensor imaging data were collected to construct brain structural networks, with both global and regional topological properties measured using graph theory. We enrolled 28 patients with early-onset schizophrenia in our study. After 5 weeks of treatment, both the intervention and active control groups showed significant improvement in psychotic symptoms, yet the improvement was greater in the intervention group. Importantly, in contrast with no brain structural network change after treatment in the active control group, the intervention group showed increased nodal centrality of the left insula that was associated with psychotic symptom improvement. We did not collect important information concerning the participants' cognitive abilities, particularly ToM performance. These findings suggest a potential neural mechanism by which ToM psychotherapy exerts a beneficial effect on early-onset schizophrenia via strengthening the coordination capacity of the insula in brain structural networks, which may provide a clinically translatable biomarker for monitoring or predicting responses to ToM psychotherapy.Clinical trial registration: NCT05577338; ClinicalTrials.gov.


Statistical tools applied in the characterisation and evaluation of a thermo-hygrometric corrective action carried out at the Noheda archaeological site (Noheda, Spain).
The Noheda archaeological site is unique and exceptional for its size, and the quality and conservation condition of the Roman mosaic pavement covering its urban pars. In 2008 a tent was installed as protection from rain and sun. Being of interest to characterise the microclimate of the remains, six probes with relative humidity and temperature sensors were installed in 2013 for this purpose. Microclimate monitoring allowed us to check relative humidity differences resulting from the groundwater level, as well as inner sensors reaching maximum temperatures higher than the outdoors ones as a consequence of the non-ventilated tent covering the archaeological site. Microclimatic conditions in the archaeological site were deemed detrimental for the conservation of the mosaics. Thus, in summer 2013, expanded clay and geotextile were installed over the mosaics as a corrective action. The outcomes of this study have proven the effectiveness of this solution to control temperature and relative humidity, helping to configure a more stable microclimate suitable for preservation of the mosaic.


Breathing pattern variability: a weaning predictor in postoperative patients recovering from systemic inflammatory response syndrome.
To investigate whether breathing pattern variability can serve as a potential weaning predictor for postoperative patients recovering from systemic inflammatory response syndrome (SIRS). A prospective measurement of retrospectively analyzed breathing pattern variability in a surgical intensive care unit. Seventy-eight mechanically ventilated SIRS patients who had undergone abdominal surgery were included when they were ready for weaning. They were divided into success (n=57) and failure (n=21) groups based upon their weaning outcome. Before weaning, tidal volume, total breath duration, inspiratory time, expiratory time, and peak inspiratory flow were continuously monitored for 30 min, while patients received 5 cmH2O pressure support weaning trial. After the patients successfully completed the trial, they were extubated. Successful weaning was defined as patients free from the ventilator for over 48 h, whereas a weaning failure was considered as reinstitution of mechanical ventilation within 48 h of extubation. The coefficient of variation and two values of standard deviation (SD1 and SD2; indicators of the dispersion of data points in the plot) obtained from the Poincaré plot of five respiratory parameters in the failure group were significantly lower than those in the success group. The area under the receiver operating characteristic curve of these variability indices was within the range of 0.73-0.80, indicating the accuracy of prediction. Small breathing pattern variability is associated with a high incidence of weaning failure in postoperative patients recovering from SIRS, and this variability may potentially serve as a weaning predictor.


Virtual Screening of Plant Volatile Compounds Reveals a High Affinity of Hylamorpha elegans (Coleoptera: Scarabaeidae) Odorant-Binding Proteins for Sesquiterpenes From Its Native Host.
Hylamorpha elegans(Burmeister) is a native Chilean scarab beetle considered to be a relevant agricultural pest to pasture and cereal and small fruit crops. Because of their cryptic habits, control with conventional methods is difficult; therefore, alternative and environmentally friendly control strategies are highly desirable. The study of proteins that participate in the recognition of odorants, such as odorant-binding proteins (OBPs), offers interesting opportunities to identify new compounds with the potential to modify pest behavior and computational screening of compounds, which is commonly used in drug discovery, may help to accelerate the discovery of new semiochemicals. Here, we report the discovery of four OBPs inH. elegans as well as six new volatiles released by its native host Nothofagus obliqua(Mirbel). Molecular docking performed between OBPs and new and previously reported volatiles from N. oblique revealed the best binding energy values for sesquiterpenic compounds. Despite remarkable divergence at the amino acid level, three of the four OBPs evaluated exhibited the best interaction energy for the same ligands. Molecular dynamics investigation reinforced the importance of sesquiterpenes, showing that hydrophobic residues of the OBPs interacted most frequently with the tested ligands, and binding free energy calculations demonstrated van der Waals and hydrophobic interactions to be the most important. Altogether, the results suggest that sesquiterpenes are interesting candidates for in vitro and in vivo assays to assess their potential application in pest management strategies.


Isolated vein thrombosis of the posterior fossa presenting as localized cerebellar venous infarctions or hemorrhages.
Cerebellar venous infarction or hemorrhage due to isolated venous thrombosis of the posterior fossa is a rare form of intracranial vein thrombosis that can be unsuspected in clinical practice. We studied 230 patients with intracranial vein thrombosis, identifying 9 (3.9%: 7 women, mean age 34 years) with neuroimaging or histopathologic evidence of localized posterior fossa vein thrombosis causing parenchymal injury limited exclusively to the cerebellum. All patients had an insidious presentation suggesting other diagnoses. Intracranial hypertension (n=6) and cerebellar (n=4) syndromes were the main clinical presentations. Intracranial vein thrombosis was idiopathic in 3 patients; associated with puerperium in 3; and with contraceptives, protein C deficiency, and dehydration in 1 case each. CT was abnormal but not diagnostic in 5 patients, showing a cerebellar hypodensity with fourth ventricle compression and variable hydrocephalus in 5 patients, and cerebellar hemorrhage in 2. Conventional MRI provided diagnosis in 6 cases, showing the causal thrombosis and cerebellar involvement; angiography was practiced in 2 of them, confirming the findings identified by MRI. In the other 3 patients, diagnosis was reached by histopathology. Thromboses were localized at the straight sinus (n=4), lateral sinuses (n=3), and superior petrosal vein (n=2). The acute case fatality rate was 22.2% (n=2), 1 (11.1%) patient was discharged in a vegetative state, 1 (11.1%) was severely disabled, and 5 (55.6%) were moderately disabled. Isolated venous thrombosis of the posterior fossa is infrequent and implies a challenging diagnosis. Risk factors for intracranial vein thrombosis and atypical cerebellar findings on CT should lead to further MRI assessment.


Fragmentation of Care After Geriatric Trauma: A Nationwide Analysis of outcomes and Predictors.
The health care system for the elderly is fragmented, that is worsened when readmission occurs to different hospitals. There is limited investigation into the impact of fragmentation on geriatric trauma patient outcomes. The aim of this study was to compare the outcomes following readmissions after geriatric trauma. The Nationwide Readmissions Database (2016-2017) was queried for elderly trauma patients (aged ≥65 years) readmitted due to any cause. Patients were divided into 2 groups according to readmission: index vs non-index hospital. Outcomes were 30 and 180-day complications, mortality, and the number of subsequent readmissions. Multivariable logistic regression was performed to analyze the independent predictors of fragmentation of care. A total of 36,176 trauma patients were readmitted, of which 3856 elderly patients (aged ≥65 years) were readmitted: index hospital (3420; 89%) vs non-index hospital (436; 11%). Following 1:2 propensity matching, elderly with non-index hospital readmission had higher rates of death and MI within 180 days (P = .01 and .02, respectively). They had statistically higher 30 and 180-day pneumonia (P < .01), CHF (P < .01), arrhythmias (P < .01), MI (P < .01), sepsis (P < .01), and UTI (P < .01). On multivariable binary logistic regression analysis, pneumonia (OR 1.70, P = .03), congestive heart failure (CHF) (OR 1.80, P = .03), female gender (OR .72, P = .04), and severe Head and Neck trauma (AIS≥3) (OR 1.50, P < .01) on index admission were independent predictors of fragmentation of care. While the increase in time to readmission (OR 1.01, P < .01) was also associated independently with non-index hospital admission. Fragmented care after geriatric trauma could be associated with higher mortality and complications.


Testbed as a RegUlatory Science Tool (TRUST): A Testbed Design for Evaluating 5G-Enabled Medical Devices.
The fifth-generation (5G) cellular communication technology introduces technical advances that can expand medical device access to connectivity services. However, assessing the safety and effectiveness of emerging 5G-enabled medical devices is challenging as relevant evaluation methods have not yet been established. In this paper, we propose a design model for 5G testbed as a regulatory science tool (TRUST) for assessing 5G connectivity enablers of medical device functions. Specifically, we first identify application specific testing needs and general testing protocols. Next, we outline the selection and customization of key system components to create a 5G testbed. A TRUST demonstration is documented through a realistic 5G testbed implementation along with the deployment of a custom-built example use-case for 5G-enabled medical extended reality (MXR). Detailed configurations, example collected data, and implementation challenges are presented. The openness of the TRUST design model allows a TRUST testbed to be easily extended and customized to incorporate available resources and address the evaluation needs of different stakeholders.


Effect of electrical stimulation on the interfacial tensile strength and amount of bone formation.
The effect of electrical stimulation upon the direct tensile strength of the interface union between porous calcium aluminate implants and bone plus the amount of bone formation were investigated in the femurs of rabbits. The study indicates there is an increased tensile strength of the interface in proportion to the amount of bone formation into the pores and the amount of electrical stimulation used "in vivo."


[Axillary artery cannulation for extracorporeal circulation].
When the ascending aorta and the femoral artery cannot be used for extracorporeal circulation, an emerging alternative is the use of axillary artery. To report the experience using the axillary artery for extracorporeal circulation. Between November 1998 and May 2002, 22 patients (14 male) were operated with extracorporeal circulation, cannulating the axillary artery. Briefly, an incision is made below the middle third of the clavicle and a cut is made on major pectoris muscle. Minor pectoris muscle is retracted and axillary artery is exposed. It is cannulated directly or with the aid of a prosthesis. Right axillary artery was used in 21 patients and in 20 it was cannulated with the aid of a prosthesis. Mean flow was 4.5 +/- 0.6 l/min. The most common indications were aortic dissection or aneurysms. The most common procedures done, were ascending aorta replacement in 8 cases and replacement of ascending aorta and aortic arch in 5. Thirty five percent of operations were emergencies and 32% were reoperations. In 15 patients (68%), a circulatory arrest was done. Of these, retrograde brain perfusion was used in 9, antegrade brain perfusion through the same axillary artery was used in 2 and mixed perfusion was used in 2. One patient had a complication related to the axillary cannulation. None had cerebrovascular accidents or thromboembolic complications. Two patients died in the postoperative period. Patients were followed up to 42 months after the procedure and no secondary complications of the cannulation were detected. When the ascending aorta and the femoral artery cannot be used, axillary artery is a good alternative for extracorporeal circulation.


Lasers in medicine and dentistry.
Since their introduction, great enthusiasm has greeted the application of lasers to medicine and dentistry. The future is exciting. Research will someday take us to the days of the "Star Trek" laser-scan tooth or bone repair procedures. Lasers are very specific in their application. Choose this new technology carefully, making choices and purchases based on quality scientific research and ongoing analysis and review.


Familial Hypercholesterolemia: The Most Frequent Cholesterol Metabolism Disorder Caused Disease.
Cholesterol is an essential component of cell barrier formation and signaling transduction involved in many essential physiologic processes. For this reason, cholesterol metabolism must be tightly controlled. Cell cholesterol is mainly acquired from two sources: Dietary cholesterol, which is absorbed in the intestine and, intracellularly synthesized cholesterol that is mainly synthesized in the liver. Once acquired, both are delivered to peripheral tissues in a lipoprotein dependent mechanism. Malfunctioning of cholesterol metabolism is caused by multiple hereditary diseases, including Familial Hypercholesterolemia, Sitosterolemia Type C and Niemann-Pick Type C1. Of these, familial hypercholesterolemia (FH) is a common inherited autosomal co-dominant disorder characterized by high plasma cholesterol levels. Its frequency is estimated to be 1:200 and, if untreated, increases the risk of premature cardiovascular disease. This review aims to summarize the current knowledge on cholesterol metabolism and the relation of FH to cholesterol homeostasis with special focus on the genetics, diagnosis and treatment.


Characteristics of Kcnn4 channels in the apical membranes of an intestinal epithelial cell line.
Intermediate-conductance K(+) (Kcnn4) channels in the apical and basolateral membranes of epithelial cells play important roles in agonist-induced fluid secretion in intestine and colon. Basolateral Kcnn4 channels have been well characterized in situ using patch-clamp methods, but the investigation of Kcnn4 channels in apical membranes in situ has been hampered by a layer of mucus that prevents seal formation. In the present study, we used patch-clamp methods to characterize Kcnn4 channels in the apical membrane of IEC-18 cells, a cell line derived from rat small intestine. A monolayer of IEC-18 cells grown on a permeable support is devoid of mucus, and tight junctions enable selective access to the apical membrane. In inside-out patches, Ca(2+)-dependent K(+) channels observed with iberiotoxin (a Kcnma1/large-conductance, Ca(2+)-activated K(+) channel blocker) and apamin (a Kcnn1-3/small-conductance, Ca(2+)-activated K(+) channel blocker) present in the pipette solution exhibited a single-channel conductance of 31 pS with inward rectification. The currents were reversibly blocked by TRAM-34 (a Kcnn4 blocker) with an IC(50) of 8.7 ± 2.0 μM. The channels were not observed when charybdotoxin, a peptide inhibitor of Kcnn4 channels, was added to the pipette solution. TRAM-34 was less potent in inhibiting Kcnn4 channels in patches from apical membranes than in patches from basolateral membranes, which was consistent with a preferential expression of Kcnn4c and Kcnn4b isoforms in apical and basolateral membranes, respectively. The expression of both isoforms in IEC-18 cells was confirmed by RT-PCR and Western blot analyses. This is the first characterization of Kcnn4 channels in the apical membrane of intestinal epithelial cells.


[Morphogenesis and course of renal amyloidosis].
Based on an analysis of the clinicolaboratory findings and morphological examination of 93 patients with renal amyloidosis the proteinuric stage was diagnosed in 24, the nephrotic in 35, and the azotemic in 34 patients. Elevation of the arterial blood pressure was revealed in 40% of the patients. During the first year since proteinuria was diagnosed, the nephrotic syndrome developed in 50% of the patients; renal failure in 75% of cases developed over 9 years. The 5-year survival since proteinuria was diagnosed amounted to 72%, the 10-year survival to 34%. The 5-year survival among patients with the nephrotic syndrome constituted 49%, whereas the 10-year survival was attained only in single cases. One year after blood creatinine became elevated only 30% of the patients were still alive. As the disease progressed, there was a gradual increase in amyloid impregnation of the glomeruli alone, while amyloidosis of the tubules was most pronounced in the nephrotic stage. No complete correlation was found between the gravity of the morphological picture of glomerular involvement and blood creatinine level which is undoubtedly influenced by alterations in the exoglomerular structures.


Speckle reduction in laser projection using a dynamic deformable mirror.
Despite of much effort and significant progress in recent years, speckle removal is still a challenge for laser projection technology. In this paper, speckle reduction by dynamic deformable mirror was investigated. Time varying independent speckle patterns were generated due to the angle diversity introduced by the dynamic mirror, and these speckle patterns were averaged out by the camera or human eyes, thus reducing speckle contrast in the final image. The speckle reduction by the wavelength diversity of the lasers was also studied. Both broadband lasers and narrowband laser were used for experiment. It is experimentally shown that speckle suppression can be attained by the widening of the spectrum of the lasers. Lower speckle contrast reduction was attained by the wavelength diversity for narrowband laser compared to the broadband lasers. This method of speckle reduction is suitable in laser projectors for wide screen applications where high power laser illumination is needed.


Anatomical Venous Variants in Children With Cerebral Sinovenous Thrombosis.
Background and Purpose- Literature is sparse on the frequency and significance of anatomical venous variants (AVVs) in pediatric cerebral sinovenous thrombosis (CSVT). Methods- We retrospectively reviewed children with CSVT and controls undergoing computed tomography/magnetic resonance venography from January 2008 to 2014. Clinical features examined included raised intracranial pressure, risk factors, and treatment. Radiological features examined included CSVT location, presence and type of AVVs, hemorrhagic venous infarction, and venous collateralization. Clinical outcome was measured by the pediatric stroke outcome measure and radiological outcome by thrombus recanalization. Results- Fifty-one children with CSVT were identified. Twenty-two (43%) had AVVs at presentation. Nineteen (86%) had hypoplasia/absence of major dural sinus, 5 (23%) had persistent fetal structures, 3 (14%) had duplications/fenestrations, and 1 (5%) had disconnected superficial and deep venous systems. Controls had a slightly higher but nonsignificant prevalence 26 (51%) of AVVs. No significant clinical and radiological differences were observed between children with CSVT and AVVs compared with those with typical venous anatomy. Conclusions- AVVs are seen in many children with and without CSVT and do not seem to alter the presentation or clinical course. The influence of these variations on the brain's ability to tolerate venous congestion because of thrombosis merits further study.


The Post-Stroke Depression Rating Scale: a test specifically devised to investigate affective disorders of stroke patients.
Owing to the lack of instruments specifically constructed to study emotional and affective disorders of stroke patients, the nature of post-stroke depression (PSD) remains controversial. With this in mind, the authors constructed a new scale, the Post-Stroke Depression Scale (PSDS) which takes into account a series of symptoms and problems commonly observed in depressed stroke patients. The PSDS and the Hamilton Depression Rating Scale (HDS) were administered to a group of 124 patients, who had been classified, on the basis of DSM III-R diagnostic criteria, in the following categories: No depression (n = 32); Minor PSD (n = 47); Major PSD (n = 45). Scores obtained by these stroke patients on the PSDS and on the HDS were compared to those obtained on the same scales by 17 psychiatric patients also classified as major depression on the basis of DSM III-R diagnostic criteria. An analysis of the symptomatological profiles clearly showed that: (1) a continuum exists between the so-called "major" and "minor" forms of PSD; (2) in both groups of depressed stroke patients the depressive symptomatology seems due to the psychological reaction to the devastating consequences of stroke, since the motivated aspects of depression prevailed in depressed stroke patients, whereas the (biologically determined) unmotivated aspects prevailed in patients with a functional form of major depression; and (3) in stroke patients a DSM III-based diagnosis of major PSD could be in part inflated by symptoms (such as apathy and vegetative disorders) that are typical of major depression in a patient free from brain damage, but that could be due to the brain lesion per se in a stroke patient.


Cellular zinc fluxes and the regulation of apoptosis/gene-directed cell death.
The maintenance of discrete subcellular pools of zinc (Zn) is critical for the functional and structural integrity of cells. Among the important biological processes influenced by Zn is apoptosis, a process that is important in cellular homeostasis (an important cellular homeostatic process). It has also been identified as a major mechanism contributing to cell death in response to toxins and in disease, offering hope that novel therapies that target apoptotic pathways may be developed. Because Zn levels in the body can be increased in a relatively nontoxic manner, it may be possible to prevent or ameliorate degenerative disorders that are associated with high rates of apoptotic cell death. This review begins with brief introductions that address, first, the cellular biology of Zn, especially the critical labile Zn pools, and, second, the phenomenon of apoptosis. We then review the evidence relating Zn to apoptosis and address three major hypotheses: (1) that a specific pool or pools of intracellular labile Zn regulates apoptosis; (2) that systemic changes in Zn levels in the body, due to dietary factors, altered physiological states or disease, can influence cell susceptibility to apoptosis, and (3) that this altered susceptibility to apoptosis contributes to pathophysiological changes in the body. Other key issues are the identity of the molecular targets of Zn in the apoptotic cascade, the types of cells and tissues most susceptible to Zn-regulated apoptosis, the role of Zn as a coordinate regulator of mitosis and apoptosis and the apparent release of tightly bound intracellular pools of Zn during the later stages of apoptosis. This review concludes with a section highlighting areas of priority for future studies.


Management of Urothelial Bladder Cancer in Clinical Practice: Real-World Answers to Difficult Questions.
Management of urothelial bladder cancer has historically been challenging as a result of a limited grasp of disease biology and few available systemic therapy options, mainly consisting of platinum-based chemotherapy. Improved understanding of molecular mechanisms underlying pathogenesis of muscle-invasive bladder cancer as well as their correlation with tumor behavior and response to treatment has emerged over the past few years. Remarkable therapeutic advances have been made with the introduction of checkpoint inhibitors, which have changed the course of this disease. Multiple agents with novel mechanisms of action are also actively being explored in ongoing clinical trials. These advances are exciting but may prove challenging in terms of how to apply this constantly evolving plethora of data to actual patients. This review addresses the gray areas and challenging questions that frequently arise in clinical practice.


Cost Benefit of Implementation of Risk Stratification Models for Adult Spinal Deformity Surgery.
Retrospective cohort study. Assess the extent to which defined risk factors of adverse events are drivers of cost-utility in spinal deformity (ASD) surgery. ASD patients with 2-year (2Y) data were included. Tertiles were used to define high degrees of frailty, sagittal deformity, blood loss, and surgical time. Cost was calculated using the Pearl Diver registry and cost-utility at 2Y was compared between cohorts based on the number of risk factors present. Statistically significant differences in cost-utility by number of baseline risk factors were determined using ANOVA, followed by a generalized linear model, adjusting for clinical site and surgeon, to assess the effects of increasing risk score on overall cost-utility. By 2 years, 31% experienced a major complication and 23% underwent reoperation. Patients with ≤2 risk factors had significantly less major complications. Patients with 2 risk factors improved the most from baseline to 2Y in ODI. Average cost increased by $8234 per risk factor (R2 = .981). Cost-per-QALY at 2Y increased by $122,650 per risk factor (R2 = .794). Adjusted generalized linear model demonstrated a significant trend between increasing risk score and increasing cost-utility (r2 = .408, P < .001). The number of defined patient-specific and surgical risk factors, especially those with greater than two, were associated with increased index surgical costs and diminished cost-utility. Efforts to optimize patient physiology and minimize surgical risk would likely reduce healthcare expenditures and improve the overall cost-utility profile for ASD interventions.Level of evidence: III.


Effect of nonuniform muscle contraction on sustainability and frequency of triggered arrhythmias in rat cardiac muscle.
Arrhythmias are benign or lethal, depending on their sustainability and frequency. To determine why lethal arrhythmias are prone to occur in diseased hearts, usually characterized by nonuniform muscle contraction, we investigated the effect of nonuniformity on sustainability and frequency of triggered arrhythmias. Force, membrane potential, and intracellular Ca(2+) concentration ([Ca(2+)](i)) were measured in 51 rat ventricular trabeculae. Nonuniform contraction was produced by exposing a restricted region of muscle to a jet of 20 mmol/L 2,3-butanedione monoxime (BDM) or 20 mumol/L blebbistatin. Sustained arrhythmias (>10 seconds) could be induced by stimulus trains for 7.5 seconds only with the BDM or blebbistatin jet (100 nmol/L isoproterenol, 1.0 mmol/L [Ca(2+)](o), 24 degrees C). During sustained arrhythmias, Ca(2+) surges preceded synchronous increases in [Ca(2+)](i), whereas the stoppage of the BDM jet made the Ca(2+) surges unclear and arrested sustained arrhythmias (n=6). With 200 nmol/L isoproterenol, 2.5 mmol/L [Ca(2+)](o), and the BDM jet, lengthening or shortening of the muscle during sustained arrhythmias accelerated or decelerated their cycle in both the absence (n=10) and presence (n=10) of 100 mumol/L streptomycin, a stretch-activated channel blocker, respectively. The maximum rate of force relaxation correlated inversely with the change in cycle lengths (n=14; P<0.01). Sustained arrhythmias with the BDM jet were significantly accelerated by 30 mumol/L SCH00013, a Ca(2+) sensitizer of myofilaments (n=10). These results suggest that nonuniformity of muscle contraction is an important determinant of the sustainability and frequency of triggered arrhythmias caused by the surge of Ca(2+) dissociated from myofilaments in cardiac muscle.


Identification of a novel function of Id-1 in mediating the anticancer responses of SAMC, a water-soluble garlic derivative, in human bladder cancer cells.
Studies have shown that the expression of inhibitor of differentiation (Id-1) is increased in bladder cancer and is associated with drug resistance. S-allylmercaptocysteine (SAMC), a water-soluble component of garlic, is known to have a potent therapeutic effect on human cancer. The aim of this study was to investigate whether Id-1 expression mediates SAMC-induced cell death in bladder cancer cells. After generating stable Id-1-expressing and si-Id-1 transfectants in various bladder cancer cell lines, cell sensitivity to SAMC was compared by colony formation and MTT assays. The results indicated that Id-1 overexpression reduced the positive effect of SAMC on cell survival, while the inactivation of Id-1 increased cellular susceptibility to SAMC. Using DAPI staining, the apoptosis of bladder cancer cells induced by SAMC was shown to be negatively regulated by Id-1 expression. The expression of apoptosis-related proteins analyzed by Western blotting further supported the negative role of Id-1 in SAMC-induced apoptosis. Furthermore, by wound closure and type I collagen invasion assays, the inhibitory effect of SAMC on the invasion and migration of bladder cancer cells was found to be associated with the down-regulation of Id-1. Our results demonstrated that SAMC-induced apoptosis is associated with the Id-1 pathway, and that the inactivation of Id-1 enhances the ability of SAMC to inhibit the survival, invasion and migration of bladder cancer cells. These findings may lead to the development of novel therapeutic strategies for the treatment of bladder cancer.


[Comparative x-ray and nuclear medical studies of osteosarcomas to evaluate the effectiveness of preoperative chemotherapy].
In 16 patients with a long bone osteosarcoma treated after the therapy regimen of COSS 80/82 (Cooperative Osteosarcoma Study) the tumour response to preoperative chemotherapy was assessed by conventional roentgenograms (plain films, tomograms, angiograms). The histological grade of regression determined in the operative specimen was used as a reference for the extent of tumour devitalisation. By their different typical roentgenological course the osteosarcomas with a very good and those with a rather poor response could be correctly identified (100%). On following the clinically used differentiation between "good responders" (histological grades I to III) and "poor responders" (grades IV to VI) the corresponding classification of these tumours by roentgenograms and functional scintigraphic imaging was found to be correct in 94% of the cases. The coincidence of the correct as well as the false results in both radiological methods was as high as 100% if those osteosarcomas showing only a "medium response" in their roentgenograms were added to the clinical group of good responders. There were no fundamentally discrepant results obtained by each of the two radiological techniques. With the combined workup of the roentgenological and the scintigraphic material any incorrect preoperative estimation of tumour regression with a harmful influence on therapy (date and type of surgery, mode of postoperative chemotherapy) could be avoided.


A meta-analysis of the efficacy of ocular prophylactic agents used for the prevention of gonococcal and chlamydial ophthalmia neonatorum.
Neonatal eye prophylaxis has been routine in North America for more than a century. Contextual changes justify reexamining this practice, and prompted a systematic review of the efficacy of prophylactic agents. We searched MEDLINE (1966-2008), EMBASE (1980-2008), CINAHL (1982-2008), and the Cochrane library (the first quarter of 2008) for relevant clinical trials and hand-searched the resulting reference lists. We independently evaluated eligibility and study quality. Meta-analyses were performed using a random effects model. Each of the eight included studies had substantial methodologic weaknesses. Data to estimate the efficacy of prophylaxis in the prevention of gonococcal ophthalmia neonatorum (GON) were not available. One study found no differences in rates of chlamydial ophthalmia neonatorum (CON) when three agents were compared to no prophylaxis: silver nitrate (relative risk [RR] = 1.06; 95% confidence interval [CI], 0.55-2.02; 2225 newborns), erythromycin (RR = 0.93; 95% CI, 0.48-1.79; 2306 newborns), and tetracycline (RR = 0.82; 95% CI, 0.42-1.63; 2299 newborns). No statistically significant differences were found between agents in the prevention of GON. Erythromycin and povidone-iodine both decrease the risk of CON when compared to silver nitrate (RR = 0.71; 95% CI, 0.52-0.97; 4514 newborns, and RR = 0.52; 95% CI, 0.38-0.71; 2005 newborns, respectively). Failure rates of universal eye prophylaxis support reexamination of this policy where the prevalence of maternal infection is low.


Adjuvanticity and strains of mycobacteria.
Water-in-oil emulsions containing mycobacteria constitute Freund's complete adjuvants (FCA), used to enhance immune responses. Mycobacteria are the essential constituent of FCA and various strains of these bacillus were used. The present study was to try to answer the follow question: Are there any differences between:-- water-in-oil emulsions containing mycobacteria of the bovine attenuated "BCG" strain, called Be (bovine emulsions) and--those containing mycobacteria of the human virulent "Canetti" strain, He (human emulsions)? Data presented here suggest that: 1. Bovine emulsions are active at low doses while human emulsions are only active at high doses; 2. The adjuvant activity of the bovine emulsions at low doses is attributed to both, the better stimulatary effect and the high concentration of an active component located in BCG cell envelope. Immuno-chemical studies of various extracts were conducted and the relationship between adjuvanticity and chemical structure of the active components was established.


Saturation mutagenesis charts the functional landscape of Salmonella ProQ and reveals a gene regulatory function of its C-terminal domain.
The global RNA-binding protein ProQ has emerged as a central player in post-transcriptional regulatory networks in bacteria. While the N-terminal domain (NTD) of ProQ harbors the major RNA-binding activity, the role of the ProQ C-terminal domain (CTD) has remained unclear. Here, we have applied saturation mutagenesis coupled to phenotypic sorting and long-read sequencing to chart the regulatory capacity of Salmonella ProQ. Parallel monitoring of thousands of ProQ mutants allowed mapping of critical residues in both the NTD and the CTD, while the linker separating these domains was tolerant to mutations. Single amino acid substitutions in the NTD associated with abolished regulatory capacity strongly align with RNA-binding deficiency. An observed cellular instability of ProQ associated with mutations in the NTD suggests that interaction with RNA protects ProQ from degradation. Mutation of conserved CTD residues led to overstabilization of RNA targets and rendered ProQ inert in regulation, without affecting protein stability in vivo. Furthermore, ProQ lacking the CTD, although binding competent, failed to protect an mRNA target from degradation. Together, our data provide a comprehensive overview of residues important for ProQ-dependent regulation and reveal an essential role for the enigmatic ProQ CTD in gene regulation.


[Integrated health information system based on Resident Assessment Instruments].
The paper explores the meaning of Resident Assessment Instruments. It gives a summary of existing RAI instruments and derived applications. It argues how all of these form the basis for an integrated health information system for "chain care" (home care, home for the elderly care, nursing home care, mental health care and acute care). The primary application of RAI systems is the assessment of client care needs, followed by an analysis of the required and administered care with the objective to make an optimal individual care plan. On the basis of RAI, however, applications have been derived for reimbursement systems, quality improvement programs, accreditation, benchmarking, best practice comparison and care eligibility systems. These applications have become possible by the development on the basis of the Minimum Data Set of RAI of outcome measures (item scores, scales and indices), case-mix classifications and quality indicators. To illustrate the possibilities of outcome measures of RAI we present a table and a figure with data of six Dutch nursing homes which shows how social engagement is related to ADL and cognition. We argue that RAI/MDS assessment instruments comprise an integrated health information system because they have consistent terminology, common core items, and a common conceptual basis in a clinical approach that emphasizes the identification of functional problems.


Constituents of the Roots of Dichapetalum pallidum and Their Anti-Proliferative Activity.
As part of our search for bioactive compounds from the Dichapetalaceae, repeated chromatographic purification of the roots of a hitherto unexamined species, Dichapetalum pallidum, led to the isolation of the newly occurring 7-hydroxydichapetalin P (1) and the known dichapetalins A (2) and X (3). Also isolated were the known compounds friedelin-2,3-lactone (4), friedelan-3-one (6), friedelan-3β-ol (7) and pomolic (8), as well as the dipeptide aurantiamide acetate (5). The compounds were characterized by direct interpretation of their IR, 1D NMR and 2D NMR spectral data and by comparison of their physico-chemical data, including their chromatographic profiles, with the literature and authentic samples in our compound library for the genus Dichapetalum. The compounds were assayed for their anti-proliferative activities against the human T-lymphocytic leukemia (Jurkat), acute promyelocytic leukemia (HL-60) and T-lymphoblast-like leukemia (CEM) cell lines. Overall, dichapetalin X showed the strongest (3.14 μM) and broadest cytotoxic activities against all the leukemic cell lines tested, exhibiting even stronger activities than the standard compound, curcumin.


Risk factors for disability pension in a population-based cohort of men and women on long-term sick leave in Sweden.
Knowledge on predictors of disability pension is very limited. The aim was to assess the importance of sick-leave diagnosis and socio-demographic variables as risk factors for disability pension among individuals on long-term sickness absence and to compare these factors by gender and over time. A prospective population-based cohort study in Ostergötland County, Sweden, included 19,379 individuals who, in 1985-87, were aged 16-60 years and had a new spell of long-term sickness absence lasting > or =56 days. Follow-up was done in two time frames: 0-5 and 6-10 years after inclusion. The risk of disability pension in relation to sick-leave diagnosis and socio-demographic factors was assessed by Cox proportional hazard regression analysis. In 5 years, after inclusion, 28% of the cohort had been granted disability pension. Those with higher age, low income, previous sick leave, no employment and non-Swedish origin had higher risk of disability pension, while those with young children had lower risk. Considering the inclusion diagnosis, the pattern differed between men and women (P < 0.001). Among men, those with mental disorders had the highest risk and among women those with musculoskeletal disorders. Except for income, the effect of which was reversed over time, the overall pattern of disability pension predictors remained 6-10 years after inclusion but was attenuated. Besides socio-demographic risk factors, the sick-leave diagnoses constitute an important both medium and long-term predictor of disability pension among both men and women on long-term sickness absence.


Single-cell genomics reveals features of a Colwellia species that was dominant during the Deepwater Horizon oil spill.
During the Deepwater Horizon (DWH) oil spill in the Gulf of Mexico a deep-sea hydrocarbon plume developed resulting in a rapid succession of bacteria. Colwellia eventually supplanted Oceanospirillales, which dominated the plume early in the spill. These successional changes may have resulted, in part, from the changing composition and abundance of hydrocarbons over time. Colwellia abundance peaked when gaseous and simple aromatic hydrocarbons increased, yet the metabolic pathway used by Colwellia in hydrocarbon disposition is unknown. Here we used single-cell genomics to gain insights into the genome properties of a Colwellia enriched during the DWH deep-sea plume. A single amplified genome (SAG) of a Colwellia cell isolated from a DWH plume, closely related (avg. 98% 16S rRNA gene similarity) to other plume Colwellia, was sequenced and annotated. The SAG was similar to the sequenced isolate Colwellia psychrerythraea 34H (84% avg. nucleotide identity). Both had genes for denitrification, chemotaxis, and motility, adaptations to cold environments and a suite of nutrient acquisition genes. The Colwellia SAG may be capable of gaseous and aromatic hydrocarbon degradation, which contrasts with a DWH plume Oceanospirillales SAG which encoded non-gaseous n-alkane and cycloalkane degradation pathways. The disparate hydrocarbon degradation pathways are consistent with hydrocarbons that were abundant at different times in the deep-sea plume; first, non-gaseous n-alkanes and cycloalkanes that could be degraded by Oceanospirillales, followed by gaseous, and simple aromatic hydrocarbons that may have been degraded by Colwellia. These insights into the genomic properties of a Colwellia species, which were supported by existing metagenomic sequence data from the plume and DWH contaminated sediments, help further our understanding of the successional changes in the dominant microbial players in the plume over the course of the DWH spill.


Schisandrin B promotes Foxp3+ regulatory T cell expansion by activating heme oxygenase-1 in dendritic cells and exhibits immunomodulatory effects in Th2-mediated allergic asthma.
Allergic asthma is induced by T helper 2 (Th2) responses and allergen-specific immunoglobulin E (IgE). In asthma, regulatory T (Treg) cells play a crucial role in controlling immune homeostasis, and induction of Treg cells is a good strategy to treat Th2-mediated allergic asthma. Schisandrin B (Sch B), the main component isolated from Schisandra chinensis, reportedly possesses various pharmacological properties, but its immunomodulatory mechanism in allergic asthma remains unclear. In the present study, we explored whether Sch B exerts an antiallergic effect through modifying the function of dendritic cells (DCs) to regulate T-cell polarization and further investigated the immunomodulatory effects of Sch B in allergic asthma. Herein, an in vitro study revealed that 20 μM of Sch B-treated bone-marrow-derived DCs exhibited a semi-mature phenotype that secreted low amounts of proinflammatory cytokines including interleukin (IL)-12, IL-1β, IL-6, and tumor necrosis factor (TNF)-α, and expressed decreased levels of surface molecules of cluster of differentiation 80 (CD80) and CD86. Compared to fully mature DCs, these Sch B-treated DCs displayed a regulatory ability to promote CD4+Foxp3+ Treg cell generation via upregulation of heme oxygenase (HO)-1 expression. Of note, in a murine model of ovalbumin (OVA)-induced asthma, levels of Th2-type cytokines such as IL-4, IL-5, and IL-13, and C-C motif chemokine 11 (CCL11) were dampened, whereas numbers of forkhead box P3 (Foxp3)-positive Treg cells were augmented in Sch B-treated mice. Moreover, administration of 5 mg/kg of Sch B alleviated the cardinal features of Th2-mediated allergic asthma, namely, serum OVA-specific IgE production, the development of airway hyperresponsiveness (AHR), and airway inflammation. Collectively, these findings indicate that the effectiveness of Sch B treatment against Th2-mediated allergic asthma was at least partially due to enhancement of DC induction of Treg cells, and Sch B can possibly be developed as an immunomodulatory adjuvant to treat allergic asthma.


Tubulin-nucleotide interactions. Effects of removal of exchangeable guanine nucleotide on protein conformation and microtubule assembly.
The removal of tightly bound GDP from the exchangeable nucleotide-binding site of tubulin has been performed with alkaline phosphatase under conditions which essentially retain the assembly properties of the protein. When microtubule protein is treated with alkaline phosphatase, nucleotide is selectively removed from tubulin dimer rather than from MAP (microtubule-associated protein)-containing oligomeric species. Tubulin devoid of E-site (the exchangeable nucleotide-binding site of the tubulin dimer) nucleotide shows enhanced proteolytic susceptibility of the beta-subunit to thermolysin and decreased protein stability, consistent with nucleotide removal causing changes in protein tertiary structure. Pyrophosphate ion (3 mM) is able to promote formation of normal microtubules in the complete absence of GTP by incubation at 37 degrees C either with nucleotide-depleted microtubule protein or with nucleotide-depleted tubulin dimer to which MAPs have been added. The resulting microtubules contain up to 80% of tubulin lacking E-site nucleotide. In addition to its effects on nucleation, pyrophosphate competes weakly with GDP bound at the E-site. It is deduced that binding of pyrophosphate at a vacant E-site can promote microtubule assembly. The minimum structural requirement for ligands to induce tubulin assembly apparently involves charge neutralization at the E-site by bidentate ligation, which stabilizes protein domains in a favourable orientation for promoting the supramolecular protein-protein interactions involved in microtubule formation.


Azotobacter vinelandii siderophore can provide nitrogen to support the culture of the green algae Neochloris oleoabundans and Scenedesmus sp. BA032.
Microalgae are viewed as a potential future agricultural and biofuel feedstock and also provide an ideal biological means of carbon sequestration based on rapid growth rates and high biomass yields. Any potential improvement using high-yield microalgae to fix carbon will require additional fertilizer inputs to provide the necessary nitrogen required for protein and nucleotide biosynthesis. The free-living diazotroph Azotobacter vinelandii can fix nitrogen under aerobic conditions in the presence of reduced carbon sources such as sucrose or glycerol and is also known to produce a variety of siderophores to scavenge different metals from the environment. In this study, we identified two strains of green algae, Neochloris oleoabundans and Scenedesmus sp. BA032, that are able to utilize the A. vinelandii siderophore azotobactin as a source of nitrogen to support growth. When grown in a co-culture, S. sp. BA032 and N. oleoabundans obtained the nitrogen required for growth through the association with A. vinelandii. These results, indicating a commensalistic relationship, provide a proof of concept for developing a mutualistic or symbiotic relationship between these two species using siderophores as a nitrogen shuttle and might further indicate an additional fate of siderophores in the environment.


[The main approaches to reducing cardiovascular morbidity and mortality in Russia].
Analysis of medical statistical data and of the socioeconomic situation in the country brought the authors to a conclusion that at present the principal factors responsible for cardiovascular diseases, specifically, coronary disease and cerebrovascular disorders, are environmental factors, such as chronic stress, alcoholism and hard drinking, violation of social and hygienic norms of labor, and a number of factors related to medicine: no prophylactic check-ups and unavailability of effective drugs. "Internal risk factors" should not be disregarded as well: hypercholesterolemia, arterial hypertension, etc. Based on their results, the authors propose measures aimed at limitation of overall stress exposures, alcoholism and hard drinking control, sociohygienic labor protection, re-creation of a system of effective prophylactic check-ups, and making present-day effective drugs available for the population.


Phase-sensitive optical low-coherence reflectometry for the detection of analyte concentrations.
Optical techniques may potentially be used for noninvasive glucose sensing. We investigated the application of phase-sensitive optical low-coherence reflectometry (PS-OLCR) to the measurement of analyte concentrations. The dependence of the PS-OLCR signal on the concentration of various analytes, including aqueous solutions of glucose, calcium chloride, magnesium chloride, sodium chloride, potassium chloride, potassium bicarbonate, urea, bovine serum albumin, and bovine globulin, were determined in clear and turbid media. Obtained results demonstrated (1) a high degree of sensitivity and accuracy of the phase measurements of analyte concentrations with PS-OLCR; (2) a concentration-dependent change in the phase-shift for glucose that is significantly greater than that of other analytes sampled over the same physiological range; and (3) a high submillimolar sensitivity of PS-OLCR for the measurement of glucose concentration. Further exploration of the application of PS-OLCR to the noninvasive, sensitive, and specific monitoring of glucose concentration seems warranted.


Rapid Generation of Neutralizing Antibody Responses in COVID-19 Patients.
SARS-CoV-2, the virus responsible for COVID-19, is causing a devastating worldwide pandemic, and there is a pressing need to understand the development, specificity, and neutralizing potency of humoral immune responses during acute infection. We report a cross-sectional study of antibody responses to the receptor-binding domain (RBD) of the spike protein and virus neutralization activity in a cohort of 44 hospitalized COVID-19 patients. RBD-specific IgG responses are detectable in all patients 6 days after PCR confirmation. Isotype switching to IgG occurs rapidly, primarily to IgG1 and IgG3. Using a clinical SARS-CoV-2 isolate, neutralizing antibody titers are detectable in all patients by 6 days after PCR confirmation and correlate with RBD-specific binding IgG titers. The RBD-specific binding data were further validated in a clinical setting with 231 PCR-confirmed COVID-19 patient samples. These findings have implications for understanding protective immunity against SARS-CoV-2, therapeutic use of immune plasma, and development of much-needed vaccines.


Comparison of two stroke volume variation-based goal-directed fluid therapies for supratentorial brain tumour resection: a randomized controlled trial.
The optimal volume status for neurosurgery has yet to be determined. We compared two fluid protocols based on different stroke volume variation (SVV) cut-offs for goal-directed fluid therapy (GDFT) during supratentorial brain tumour resection. A randomized, single-blind, open-label trial was conducted. Eighty adult patients undergoing elective supratentorial brain tumour resection were randomly divided into a low SVV and a high SVV group. The SVV cut-offs were used to determine when to initiate colloid infusion. Clinical outcomes and perioperative changes in serum neuronal biomarkers, including S100β, neurone-specific enolase (NSE) and glial fibrillary acidic protein (GFAP), were compared. Patients in the low SVV group received a higher volume of colloid [869 (SD 404) vs 569 (453) ml; P=0.0025], had a higher urine output [3.4 (2.4) vs 2.5 (1.7) ml kg-1 h-1; P=0.0416] and a higher average cardiac index [3.2 (0.7) vs 2.8 (0.6) litres min-1 m-2; P=0.0204]. Patients in the low SVV group also had a shorter intensive care unit stay [1.4 (0.7) vs 2.6 (3.3) days, P=0.0326], fewer postoperative neurological events (17.5 vs 40%, P=0.0469), attenuated changes in the NSE and GFAP levels, lower intraoperative serum lactate and a higher Barthel index at discharge (all P<0.05). During GDFT for supratentorial brain tumour resection, fluid boluses targeting a lower SVV are more beneficial than a restrictive protocol. NCT02113358.


Salience network dynamics underlying successful resistance of temptation.
Self-control and the ability to resist temptation are critical for successful completion of long-term goals. Contemporary models in cognitive neuroscience emphasize the primary role of prefrontal cognitive control networks in aligning behavior with such goals. Here, we use gaze pattern analysis and dynamic functional connectivity fMRI data to explore how individual differences in the ability to resist temptation are related to intrinsic brain dynamics of the cognitive control and salience networks. Behaviorally, individuals exhibit greater gaze distance from target location (e.g. higher distractibility) during presentation of tempting erotic images compared with neutral images. Individuals whose intrinsic dynamic functional connectivity patterns gravitate toward configurations in which salience detection systems are less strongly coupled with visual systems resist tempting distractors more effectively. The ability to resist tempting distractors was not significantly related to intrinsic dynamics of the cognitive control network. These results suggest that susceptibility to temptation is governed in part by individual differences in salience network dynamics and provide novel evidence for involvement of brain systems outside canonical cognitive control networks in contributing to individual differences in self-control.


Community-acquired pneumonia: Epidemiology, diagnosis, prognostic severity scales, and new therapeutic options.
This narrative review article explores the current scientific knowledge on the definition, epidemiology, diagnostic criteria, microbiology, treatment, and prevention of severe community-acquired pneumonia (SCAP) in immunocompetent adults. At present, despite major scientific advances in diagnostic evaluation, clinical management, antimicrobial therapy, and prevention, severe community-acquired pneumonia remains a major cause of morbidity and mortality, as well as having a major economic impact in terms of increased healthcare expenditure worldwide. This pathology is considered one of the leading causes of sepsis/septic shock, with an extremely high overall mortality rate, which justifies all the effort in early diagnosis, proper management, and prompt initiation of antimicrobial therapy. Including biomarkers (isolated or in combination) associated with applying diagnostic criteria and prognostic severity scales in clinical practice helps identify patients with severe community-acquired pneumonia, defines immediate admission to the intensive care unit, and, thus, minimizes the adverse outcomes of this serious pathology.


Evidence for reduced 3 beta-ol-hydroxysteroid dehydrogenase activity in some hirsute women thought to have polycystic ovary syndrome.
Congenital adrenal hyperplasia due to 3 beta-ol-hydroxysteroid dehydrogenase (3 beta-ol) deficiency is usually lethal. A partial deficiency in 3 beta-ol has been suggested in some women presenting with androgen excess. In this study, 24 women were investigated who had hirsutism and oligomenorrhea and high serum delta 5 androgens compared to delta 4 androgens. Of these women, 9 had significantly elevated 17-hydroxypregnenolone (17 Preg) to 17-hydroxyprogesterone (17 Prog) ratios when compared to controls under basal conditions. On further testing of 9 women with ACTH, 4 had significantly elevated 17 Preg to 17 Prog ratios. Eight women had elevated ratios of dehydroepiandrosterone sulfate to androstenedione, 4 had elevations of androstenediol (Adiol) to testosterone (T), and 4 had abnormal 17 Preg to cortisol ratios. Only 3 women out of the original 24 selected for study had elevated ratios for all 4 different steroid pairs measured. 17 Prog was normal in these women with 3 beta-ol deficiency in contradistinction to the high levels normally observed in women with congenital adrenal hyperplasia due to 21- or 11-hydroxylase deficiency. It is suggested that the cause of androgen excess in these women is high circulating levels of Adiol and, in part, the slightly elevated unbound T levels in these women. In conclusion, a subtle, incomplete form of 3 beta-ol deficiency may exist in adult women and is manifest by high delta 5 androgens such as dehydroepiandrosterone sulfate and Adiol and normal delta 4 androgens such as androstenedione and T.


Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears.
Normal blood smears were stained by the standardised azure B-eosin Y Romanowsky procedure recently introduced by the ICSH, and the classical picture resulted. The effects of varying the times and temperature of staining, the composition of the solvent (buffer concentration, methanol content, & pH), the concentration of the dyes, and the mode of fixation were studied. The results are best understood in terms of the following staining mechanism. Initial colouration involves simple acid and basic dyeing. Eosin yields red erythrocytes and eosinophil granules. Azure B very rapidly gives rise to blue stained chromatin, neutrophil specific granules, platelets and ribosome-rich cytoplasms; also to violet basophil granules. Subsequently the azure B in certain structures combines with eosin to give purple azure B-eosin complexes, leaving other structures with their initial colours. The selectivity of complex formation is controlled by rate of entry of eosin into azure B stained structures. Only faster staining structures (i.e. chromatin, neutrophil specific granules, and platelets) permit formation of the purple complex in the standard method. This staining mechanism illuminates scientific problems (e.g. the nature of 'toxic' granules) and assists technical trouble-shooting (e.g. why nuclei sometimes stain blue, not purple).


Investigating neurogenic bowel in experimental spinal cord injury: where to begin?
The devastating losses following traumatic spinal cord injury (SCI) encompass the motor, sensory and autonomic nervous systems. Neurogenic bowel is a slow transit colonic dysfunction marked by constipation, rectal evacuation difficulties, decreased anorectal sensation, fecal incontinence or some combination thereof. Furthermore, neurogenic bowel is one of the most prevalent comorbidities of SCI and is recognized by afflicted individuals and caregivers as a lifelong physical and psychological challenge that profoundly affects quality of life. The restoration of post-injury control of movement has received considerable scientific scrutiny yet the daily necessity of voiding the bowel and bladder remains critically under-investigated. Subsequently, physicians and caregivers are rarely presented with consistent, evidence-based strategies to successfully address the consequences of dysregulated voiding reflexes. Neurogenic bowel is commonly believed to result from the interruption of the supraspinal control of the spinal autonomic circuits regulating the colon. In this mini-review, we discuss the clinical challenges presented by neurogenic bowel and emerging pre-clinical evidence that is revealing that SCI also initiates functional remodeling of the colonic wall concurrent with a decrease in local enteric neurons. Since the enteric input to the colonic smooth muscle is the final common pathway for functional contractions of the colon, changes to the neuromuscular interface must first be understood in order to maximize the efficacy of therapeutic interventions targeting colonic dysfunction following SCI.


Synthesis, Characterization, and Simulation of Four-Armed Megamolecules.
This paper describes the synthesis, characterization, and modeling of a series of molecules having four protein domains attached to a central core. The molecules were assembled with the "megamolecule" strategy, wherein enzymes react with their covalent inhibitors that are substituted on a linker. Three linkers were synthesized, where each had four oligo(ethylene glycol)-based arms terminated in a para-nitrophenyl phosphonate group that is a covalent inhibitor for cutinase. This enzyme is a serine hydrolase and reacts efficiently with the phosphonate to give a new ester linkage at the Ser-120 residue in the active site of the enzyme. Negative-stain transmission electron microscopy (TEM) images confirmed the architecture of the four-armed megamolecules. These cutinase tetramers were also characterized by X-ray crystallography, which confirmed the active-site serine-phosphonate linkage by electron-density maps. Molecular dynamics simulations of the tetracutinase megamolecules using three different force field setups were performed and compared with the TEM observations. Using the Amberff99SB-disp + pH7 force field, the two-dimensional projection distances of the megamolecules were found to agree with the measured dimensions from TEM. The study described here, which combines high-resolution characterization with molecular dynamics simulations, will lead to a comprehensive understanding of the molecular structures and dynamics for this new class of molecules.


Antioxidant activities of two sweet pepper Capsicum annuum L. varieties phenolic extracts and the effects of thermal treatment.
Sweet peppers Capsicum annuum L. (C. annuum) are an excellent source of vitamins A and C as well as phenolic compounds, which are important antioxidant components that may reduce the risk of diseases. The objective of this study was to evaluate their antioxidant activity under various temperatures. To compare the antioxidant activity in various temperatures (20, 35, 50, and 65 °C), two different types of colored (red and green) sweet bell peppers C annuum were selected. The red peppers were selected from those cultivated in Shahreza, Esfahan and the green peppers with the local name of Gijlar were selected from those cultivated in Urmia, West Azarbayjan. The experiments were carried out to measure the total phenolic and flavonoid content, ferric reducing antioxidant power (FRAP), chain-breaking activity, scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), and hydrogen peroxide radicals. Total phenol and flavonoid contents of pepper extracts were enhanced with increasing temperature to 65 °C. Scavenging capacity of DPPH radical of red pepper extract was enhanced because of putting at 50 °C for 30 min and for Gijlar pepper extract scavenging capacity was increased at 65 °C. Scavenging capacity of hydrogen peroxide radical of extracts was the highest at 35 °C. Chain-breaking activity of red pepper extract was increased for 60 min at 35 °C. FRAP (C) of red pepper extract was significantly different (p<0.05) in compare with Gijlar pepper. An appropriate temperature maintained a high antioxidant activity of phenolic compound, which could be due to the combined effect of non enzymatic reaction and phenolic compound stability.


Distribution and predictors of exercise habits among pregnant women in the Danish National Birth Cohort.
Physical activity is recommended during pregnancy, although strong evidence on reproductive health is lacking. We present exercise habits and predictors of exercise during pregnancy. From the Danish National Birth Cohort (1996-2002), 88,200 singleton pregnancies were analyzed in logistic regression. About one-third of the women exercised in early/mid pregnancy and slightly less in late pregnancy. Bicycling, swimming, and low-impact activities were most common. Exercising more than three times per week was strongly correlated with older age, being a student or out of work, eating disorders, moderate alcohol consumption, and a healthy diet. Multiparity, a normal or less good self-rated health, smoking, and a less health conscious diet were the strongest predictors of not doing exercise. Women of 25 years or older, with metabolic or psychiatric disorders, or who had received subfecundity treatment were more likely to increase their activity level substantially from early to late pregnancy than comparison groups. In conclusion, exercising during pregnancy correlated with a number of maternal characteristics. The findings may be used to identify pregnant women not likely to exercise, to target activities that may fit their needs, and, for research purposes, to identify adjustment variables or guide sensitivity analyses when data on confounders are lacking.


Deciphering the regulatory network of the NAC transcription factor FvRIF, a key regulator of strawberry (Fragaria vesca) fruit ripening.
The NAC transcription factor ripening inducing factor (RIF) was previously reported to be necessary for the ripening of octoploid strawberry (Fragaria × ananassa) fruit, but the mechanistic basis of RIF-mediated transcriptional regulation and how RIF activity is modulated remains elusive. Here, we show that FvRIF in diploid strawberry, Fragaria vesca, is a key regulator in the control of fruit ripening and that knockout mutations of FvRIF result in a complete block of fruit ripening. DNA affinity purification sequencing coupled with transcriptome deep sequencing suggests that 2,080 genes are direct targets of FvRIF-mediated regulation, including those related to various aspects of fruit ripening. We provide evidence that FvRIF modulates anthocyanin biosynthesis and fruit softening by directly regulating the related core genes. Moreover, we demonstrate that FvRIF interacts with and serves as a substrate of MAP kinase 6 (FvMAPK6), which regulates the transcriptional activation function of FvRIF by phosphorylating FvRIF at Thr-310. Our findings uncover the FvRIF-mediated transcriptional regulatory network in controlling strawberry fruit ripening and highlight the physiological significance of phosphorylation modification on FvRIF activity in ripening.


[Prognostic value of apolipoproteins A and B in the clinical course of patients with chronic kidney disease previous to dialysis].
Dyslipidemia is a well-established risk factor for cardiovascular diseases in the general population. However, this association is not observed in chronic kidney disease (CKD) patients. This study examines the association between lipid levels, including apolipoproteins A-I and B concentrations, and all-cause mortality or the development of new cardiovascular events in advanced CKD patients not yet on dialysis. This observational prospective historical study included 331 patients with CKD stage 4 or 5 not yet on dialysis. In addition to conventional clinical and biochemical data, total cholesterol, triglycerides, HDL, LDL, apolipoprotein A-I (apo A) and B (apo B) plasma concentrations were measured. Cox proportional hazard models were adjusted for age, sex, comorbidity index, residual renal function, serum albumin, C-reactive protein levels, and treatment with statins. The median follow-up time was 985 days, and during this period 105 patients died and 54 patients had a new cardiovascular event. In fully-adjusted fixed-covariate Cox models, the hazard ratio for each 10 mg/dl increase of apo A concentration was 0.915 (C.I. 95% 0.844 to 0.992; p=0,031). Patients with an apo A /apo B ratio in the upper tertile (i.e. > 1.42) had a better survival than that of the rest of study patients (hazard ratio = 0.592, C.I. 95% 0.368 to 0.953, p<0.05). None of the study lipid parameters was associated with new cardiovascular events in the adjusted models. In conclusion, apo A concentrations and high apo A / apo B ratios added independent predictive information about survival of CKD patients not yet on dialysis.


Efficacy and safety of topical sevoflurane in the treatment of chronic skin ulcers.
Results of efficacy and safety assessments of topical sevoflurane use in patients with long-term treatment-refractory vascular ulcers are reported. Patients were randomly assigned to receive sevoflurane instillations (1 mL per cm2 of ulcer area 1-4 times daily) plus standard wound care (ulcer cleaning, debridement, and dressing changes) or standard care only. Topical sevoflurane was initiated during hospitalization, with self- or nurse-administered instillations continued after discharge. Study participants were evaluated at least once weekly for 1 month and then every 2 weeks for up to 90 days. The primary efficacy measures were debridement-related and overall pain (assessed using a 10-point visual analog scale), daily opioid use, and ulcer size; secondary measures were patient and clinician impressions of improvement and ulcer-related admissions during treatment. The primary safety endpoint was intolerable sevoflurane-related adverse effects. Compared with the group receiving standard care alone (n = 5), the sevoflurane group (n = 10) had significant (p = 0.001) reductions in mean ± S.D. scores for debridement-related pain on day 1 of treatment and at subsequent time points; the sevoflurane group also had significant reductions in overall pain, daily opioid use, and ulcer size. Outcomes in terms of patient- and clinician-rated improvement and emergency admissions also favored the sevoflurane group. Mild localized reddening in the area surrounding ulcers occurred in 4 sevoflurane-treated patients. Direct application of sevoflurane onto vascular ulcers resulted in an intense and long-lasting analgesia and was associated with a progressive reduction of ulcer size.


Pump flow estimation from pressure head and power uptake for the HeartAssist5, HeartMate II, and HeartWare VADs.
The use of long-term mechanical circulatory support (MCS) for heart failure by means of implanted continuous-flow left ventricular assist devices (cf-LVADs) will increase, either to enable recovery or to provide a destination therapy. The effectiveness and user-friendliness of MCS will depend on the development of near-physiologic control strategies for which accurate estimation of pump flow is essential. To provide means for the assessment of pump flow, this study presents pump models, estimating pump flow (Q(lvad)) from pump speed (n) and pressure difference across the LVAD (Δp(lvad)) or power uptake (P). The models are evaluated for the axial-flow LVADs HeartAssist5 (HA5) and HeartMate II (HMII), and for a centrifugal pump, the HeartWare (HW). For all three pumps, models estimating Q(lvad) from Δp(lvad) only is capable of describing pump behavior under static conditions. For the axial pumps, flow estimation from power uptake alone was not accurate. When assuming an increase in pump flow with increasing power uptake, low pump flows are overestimated in these pumps. Only for the HW, pump flow increased linearly with power uptake, resulting in a power-based pump model that estimates static pump flow accurately. The addition of pressure head measurements improved accuracy in the axial cf-LVAD estimation models.


Neuroendocrine-induced synthesis of bone marrow-derived cytokines with inflammatory immunomodulating properties.
Although cytokines and other soluble regulators of immunity are known to be involved in hematopoiesis, little is known about the signals that induce the synthesis of those mediators locally. Based on recent studies linking the neuroendocrine hormone thyrotropin [thyroid-stimulating hormone (TSH)] to immune cell function in other tissues, we investigated the capacity of TSH to activate cytokine responses from bone marrow cells. These studies reveal that stimulation of the TSH receptor on bone marrow cells-using highly purified or recombinant TSH or by direct stimulation with anti-TSH receptor antibodies-rapidly induces the synthesis of cytokines from bone marrow cells that are classically used in the regulation of inflammatory responses. Of 13 cytokines screened for activity by ELISA or by RNase protection assays for gene expression, IL-6, IFN-beta, TNFalpha, TNFbeta, TGFbeta2, and lymphotoxin-beta responses were reproducibly induced by TSH within 2-3 h of stimulation. Intracellularly, TSH stimulation of bone marrow cells caused rapid increases in cAMP levels and induced the phosphorylation of the Jak2 protein kinase, thereby defining a novel G-protein-coupled receptor/cytokine synthesis pathway. These findings demonstrate that TSH can serve as a primary inductive signal of cytokine production by bone marrow cells.