Serotypes of enterotoxigenic Escherichia coli isolated in the United States.
Strains of enterotoxigenic Escherichia coli isolated from humans in the United States were found in 11 of 16 serotypes that previously were documented in the international literature as associated with enterotoxin production. Of 68 strains belonging to these 11 serotypes, 28 (41%) were enterotoxigenic; none of 46 strains belonging to 5 other previously implicated serotypes was enterotoxigenic. Control cultures of various serotypes were selected for comparison and found to contain 0 to 7% enterotoxigenic E. coli. E. coli belonging to documented enterotoxin-associated serotypes, characterized by both O and H antigens, were selected for toxin testing to determine their prevalence and potential pathogenicity in this country. In this study, a strain possessing any combination of an enterotoxin-associated serotype O antigen and H antigen was more likely to be enterotoxigenic than strains possessing only the specific O antigen or H antigen or neither. Five E. coli strains belonging to undocumented enterotoxin-associated serotypes did contain a combination of previously reported enterotoxin-associated serotype O and H antigens and did produce enterotoxin.


Mainstream Smoke Levels of Volatile Organic Compounds in 50 U.S. Domestic Cigarette Brands Smoked With the ISO and Canadian Intense Protocols.
A significant portion of the increased risk of cancer and respiratory disease from exposure to cigarette smoke is attributed to volatile organic compounds (VOCs). In this study, 21 VOCs were quantified in mainstream cigarette smoke from 50U.S. domestic brand varieties that included high market share brands and 2 Kentucky research cigarettes (3R4F and 1R5F). Mainstream smoke was generated under ISO 3308 and Canadian Intense (CI) smoking protocols with linear smoking machines with a gas sampling bag collection followed by solid phase microextraction/gas chromatography/mass spectrometry (SPME/GC/MS) analysis. For both protocols, mainstream smoke VOC amounts among the different brand varieties were strongly correlated between the majority of the analytes. Overall, Pearson correlation (r) ranged from 0.68 to 0.99 for ISO and 0.36 to 0.95 for CI. However, monoaromatic compounds were found to increase disproportionately compared to unsaturated, nitro, and carbonyl compounds under the CI smoking protocol where filter ventilation is blocked. Overall, machine generated "vapor phase" amounts (µg/cigarette) are primarily attributed to smoking protocol (e.g., blocking of vent holes, puff volume, and puff duration) and filter ventilation. A possible cause for the disproportionate increase in monoaromatic compounds could be increased pyrolysis under low oxygen conditions associated with the CI protocol. This is the most comprehensive assessment of volatile organic compounds (VOCs) in cigarette smoke to date, encompassing 21 toxic VOCs, 50 different cigarette brand varieties, and 2 different machine smoking protocols (ISO and CI). For most analytes relative proportions remain consistent among U.S. cigarette brand varieties regardless of smoking protocol, however the CI smoking protocol did cause up to a factor of 6 increase in the proportion of monoaromatic compounds. This study serves as a basis to assess VOC exposure as cigarette smoke is a principle source of overall population-level VOC exposure in the United States.


Identification and validation of SRC and phospho-SRC family proteins in circulating mononuclear cells as novel biomarkers for pancreatic cancer.
There is an urgent need to develop novel markers of pancreatic cancer to facilitate early diagnosis. Pancreatic carcinoma is characterized by marked stroma formation with a high number of infiltrating tumor-associated macrophages (TAMs) that originate from circulating mononuclear cells (MNCs). We hypothesized that differential analysis of protein expression and phosphorylation in circulating MNCs from healthy nude mice and nude mice bearing orthotopic human pancreatic cancer would identify a surrogate marker of pancreatic cancer. These differences were analyzed by two-dimensional gel electrophoresis followed by Western blot analysis using antibody against phosphorylated tyrosine proteins (pY). Protein and phosphorylated protein spots of interest were identified by mass spectrometry and validated by Western blot analysis as candidate markers for pancreatic cancer. We found that the expression and phosphorylation of Src family proteins were significantly higher in circulating MNCs from mice bearing pancreatic cancer than in circulating MNCs from healthy mice. TAMs in mice with pancreatic tumors also had higher Src family protein expression and phosphorylation than resident macrophages in the pancreas of healthy mice. The expression and phosphorylation of Src family proteins were correlated with tumor weight; however, increased Src expression and phosphorylation also occurred in MNCs from mice with chronic pancreatitis. This is the first report to explore novel pancreatic tumor markers in circulating MNCs. Although the specificity of the marker for pancreatic cancer was low, it could be used to monitor the disease or to select high-risk patients with chronic pancreatitis.


Inhibitory cardiac transcription factor, SRF-N, is generated by caspase 3 cleavage in human heart failure and attenuated by ventricular unloading.
Knowledge about molecular mechanisms leading to heart failure is still limited, but reduced gene activities and modest activation of caspase 3 are hallmarks of end-stage heart failure. We postulated that serum response factor (SRF), a central cardiac transcription factor, might be a cleavage target for modest activated caspase 3, and this cleavage of SRF may play a dominant inhibitory role in propelling hearts toward failure. We examined SRF protein levels from cardiac samples taken at the time of transplantation in 13 patients with end-stage heart failure and 7 normal hearts. Full-length SRF was markedly reduced and processed into 55- and 32-kDa subfragments in all failing hearts. SRF was intact in normal samples. In contrast, the hearts of 10 patients with left ventricular assist devices showed minimal SRF fragmentation. Specific antibodies to N- and C-terminal SRF sequences and site-directed mutagenesis revealed 2 alternative caspase 3 cleavage sites, so that 2 fragments were detected of each containing either the N- or C-terminal SRF. Expression of SRF-N, the 32-kDa fragment, in myogenic cells inhibited the transcriptional activity of alpha-actin gene promoters by 50% to 60%, which suggests that truncated SRF functioned as a dominant-negative transcription factor. Caspase 3 activation in heart failure sequentially cleaved SRF and generated a dominant-negative transcription factor, which may explain the depression of cardiac-specific genes. Moreover, caspase 3 activation may be reversible in the failing heart with ventricular unloading.


Prevalence of cardiac abnormalities and heart failure in unselected out-patients with type 2 diabetes mellitus and associated clinical factors: Real-world evidence from an Indian registry.
Type 2 diabetes mellitus (T2DM) is known to be associated with development of left ventricular (LV) dysfunction and heart failure (HF). The study aimed to determine the prevalence of LV dysfunction and HF in unselected out-patients with T2DM with no previous cardiac history and to correlate LV dysfunction and HF with demographic and comorbid characteristics. This cross-sectional study conducted at 27 centers in India captured demographic and clinical data through electronic case record forms. B-type natriuretic peptide of >105 pg/mL was used to diagnose HF and two-dimensional echocardiography was used to assess LV dysfunction. Of the 615 patients, 54.3 % (n = 334) were males; mean age was 57.4 ± 10.48 years. More than one-third of the patients had T2DM duration of >10 years (n = 238; 38.7 %), with hypertension as the most prevalent comorbidity (n = 372, 78.6 %). Approximately 61.3 % of the patients had LV hypertrophy. The mean LV mass was 135.0 ± 56.16 g (95 % CI 130.28, 139.70). The prevalence of any type of LV dysfunction, including systolic or diastolic dysfunction and HF was 55 % (95 % CI 51.0, 59.0) and 10 % (95 % CI 7.0, 12.0), respectively. A negligible but statistically significant correlation was observed between LV dysfunction and T2DM duration (p = 0.011), alongside HF and age (p < 0.0001). Real-world data from this registry from India demonstrates a substantial burden of LV dysfunction and HF in individuals with T2DM in India. It is imperative to formulate strategies for early identification of LV dysfunction in individuals with T2DM for prevention and consequent management of HF.


Regulation of flower development in Arabidopsis by SCF complexes.
SCF complexes are the largest and best studied family of E3 ubiquitin protein ligases that facilitate the ubiquitylation of proteins targeted for degradation. The SCF core components Skp1, Cul1, and Rbx1 serve in multiple SCF complexes involving different substrate-specific F-box proteins that are involved in diverse processes including cell cycle and development. In Arabidopsis, mutations in the F-box gene UNUSUAL FLORAL ORGANS (UFO) result in a number of defects in flower development. However, functions of the core components Cul1 and Rbx1 in flower development are poorly understood. In this study we analyzed floral phenotypes caused by altering function of Cul1 or Rbx1, as well as the effects of mutations in ASK1 and ASK2. Plants homozygous for a point mutation in the AtCUL1 gene showed reduced floral organ number and several defects in each of the four whorls. Similarly, plants with reduced AtRbx1 expression due to RNA interference also exhibited floral morphological defects. In addition, compared to the ask1 mutant, plants homozygous for ask1 and heterozygous for ask2 displayed enhanced reduction of B function, as well as other novel defects of flower development, including carpelloid sepals and an inhibition of petal development. Genetic analyses demonstrate that AGAMOUS (AG) is required for the novel phenotypes observed in the first and second whorls. Furthermore, the genetic interaction between UFO and AtCUL1 supports the idea that UFO regulates multiple aspects of flower development as a part of SCF complexes. These results suggest that SCF complexes regulate several aspects of floral development in Arabidopsis.


Alkamides from Echinacea disrupt the fungal cell wall-membrane complex.
We tested the hypothesis that alkamides from Echinacea exert antifungal activity by disrupting the fungal cell wall/membrane complex. Saccharomyces cerevisiae cells were treated separately with each of seven synthetic alkamides found in Echinacea extracts. The resulting cell wall damage and cell viability were assessed by fluorescence microscopy after mild sonication. Membrane disrupting properties of test compounds were studied using liposomes encapsulating carboxyfluorescein. Negative controls included hygromycin and nourseothricin (aminoglycosides that inhibit protein synthesis), and the positive control used was caspofungin (an echinocandin that disrupts fungal cell walls). The results show that yeast cells exposed to sub-inhibitory concentrations of each of the seven alkamides and Echinacea extract exhibit increased frequencies of cell wall damage and death that were comparable to caspofungin and significantly greater than negative controls. Consistent with effects of cell wall damaging agents, the growth inhibition by three representative alkamides tested and caspofungin, but not hygromycin B, were partially reversed in sorbitol protection assays. Membrane disruption assays showed that the Echinacea extract and alkamides have pronounced membrane disruption activity, in contrast to caspofungin and other controls that all had little effect on membrane stability. A Quantitative Structure-Activity Relationship (QSAR) analysis was performed to study the effect of structural substituents on the antifungal activity of the alkamides. Among the set studied, diynoic alkamides showed the greatest antifungal and cell wall disruption activities while an opposite trend was observed in the membrane disruption assay where the dienoic group was more effective. We propose that alkamides found in Echinacea act synergistically to disrupt the fungal cell wall/membrane complex, an excellent target for specific inhibition of fungal pathogens. Structure-function relationships provide opportunities for synthesis of alkamide analogs with improved antifungal activities.


A disk of scattered icy objects and the origin of Jupiter-family comets.
Orbital integrations carried out for 4 billion years produced a disk of scattered objects beyond the orbit of Neptune. Objects in this disk can be distinguished from Kuiper belt objects by a greater range of eccentricities and inclinations. This disk was formed in the simulations by encounters with Neptune during the early evolution of the outer solar system. After particles first encountered Neptune, the simulations show that about 1 percent of the particles survive in this disk for the age of the solar system. A disk currently containing as few as approximately 6 x 10(8) objects could supply all of the observed Jupiter-family comets. Two recently discovered objects, 1996 RQ20 and 1996 TL66, have orbital elements similar to those predicted for objects in this disk, suggesting that they are thus far the only members of this disk to be identified.


Time series modeling of pertussis incidence in China from 2004 to 2018 with a novel wavelet based SARIMA-NAR hybrid model.
It is a daunting task to discontinue pertussis completely in China owing to its growing increase in the incidence. While basic to any formulation of prevention and control measures is early response for future epidemic trends. Discrete wavelet transform(DWT) has been emerged as a powerful tool in decomposing time series into different constituents, which facilitates better improvement in prediction accuracy. Thus we aim to integrate modeling approaches as a decision-making supportive tool for formulating health resources. We constructed a novel hybrid method based on the pertussis morbidity cases from January 2004 to May 2018 in China, where the approximations and details decomposed by DWT were forecasted by a seasonal autoregressive integrated moving average (SARIMA) and nonlinear autoregressive network (NAR), respectively. Then, the obtained values were aggregated as the final results predicted by the combined model. Finally, the performance was compared with the SARIMA, NAR and traditional SARIMA-NAR techniques. The hybrid technique at level 2 of db2 wavelet including a SARIMA(0,1,3)(1,0,0)12modelfor the approximation-forecasting and NAR model with 12 hidden units and 4 delays for the detail d1-forecasting, along with another NAR model with 11 hidden units and 5 delays for the detail d2-forecasting notably outperformed other wavelets, SARIMA, NAR and traditional SARIMA-NAR techniques in terms of the mean square error, root mean square error, mean absolute error and mean absolute percentage error. Descriptive statistics exhibited that a substantial rise was observed in the notifications from 2013 to 2018, and there was an apparent seasonality with summer peak. Moreover, the trend was projected to continue upwards in the near future. This hybrid approach has an outstanding ability to improve the prediction accuracy relative to the others, which can be of great help in the prevention of pertussis. Besides, under current trend of pertussis morbidity, it is required to urgently address strategically within the proper policy adopted.


Ethenolysis of methyl oleate in room-temperature ionic liquids.
Unsaturated vegetable oils are an attractive renewable feedstock, and the selective cleavage of unsaturated fatty esters is an important transformation in this respect. The efficient and selective cross-metathesis of methyl oleate with ethylene was achieved under mild conditions with ruthenium-alkylidene catalysts in toluene and room-temperature ionic liquids (RTILs) to give two important chemical intermediates, 1-decene and methyl 9-decenoate, without double-bond migration. As recovery of the catalyst is an important target with respect to industrial applications, catalyst recycling studies were also carried out in RTILs with the first-generation Hoveyda catalyst.


Differences in prolactin and LH responses to acute stress between peripuberal and adult male rats.
Pituitary-adrenal, pituitary-gonadal and prolactin responses to acute stress (restraint) were studied in peripuberal and adult male rats. The pituitary-adrenal response to restraint stress did not differ in peripuberal and adult rats. Prolactin increase during stress was less marked in peripuberal animals. While an increase in LH during stress was observed in adult rats, peripuberal animals did not respond to stress. Testosterone levels were also lower in peripuberal than in adult rats. Diminished LH and prolactin responses to stress in peripuberal rats did not appear to be due either to increased pituitary-adrenal activity or to altered pituitary responsiveness to LHRH and dopamine respectively. Peripuberal rats were also more sensitive to the action of morphine on LH and prolactin release than were adult rats, suggesting that endogenous opioids may be involved in the LH and prolactin responses to acute stress. Differences in the maturation of central mechanisms rather than in pituitary response appear to be responsible for the differing responses to acute stress.


First structure of protein kinase CK2 catalytic subunit with an effective CK2β-competitive ligand.
The constitutively active Ser/Thr kinase CK2 (casein kinase 2) is used by tumor cells to acquire apoptosis resistance. CK2 exists as a heterotetrameric holoenzyme with two catalytic chains (CK2α) attached to a dimer of noncatalytic subunits (CK2β). A druggable cavity at the CK2β interface of CK2α allows the design of small molecules disturbing the CK2α/CK2β interaction and thus affecting activity, stability, and substrate specificity. We describe here the first structure of CK2α with an effective CK2β-competitive compound, namely, a 13-meric cyclic peptide derived from the C-terminal CK2β segment. Some well-ordered water molecules not visible in CK2 holoenzyme structures were detected at the interface. Driven mainly by enthalpy, the peptide binds with submicromolar affinity to CK2α, stimulates its catalytic activity, and reduces effectively the CK2α/CK2β affinity. The results provide a thermodynamic and structural rationalization of the peptide's CK2β-competitive functionality and pave thus the way to a peptidomimetic drug addressing the CK2α/CK2β interaction.


A phase I study of monohydroxyethylrutoside in healthy volunteers.
The flavonol monohydroxyethylrutoside (monoHER) has demonstrated protection against doxorubicin-induced cardiotoxicity in in vitro and in vivo studies without affecting the antitumor effect. In the present phase I study, the possible side effects and the pharmacokinetics of monoHER were evaluated in healthy volunteers with the aim to develop a safe and feasible dose to be evaluated in cancer patients treated with doxorubicin. The study was performed as a single blind, randomized trial in healthy volunteers (age between 19 and 56 years). At each dose level, six subjects received monoHER and three placebo. MonoHER was solubilized in 100 ml dextrose 5% and administered as an i.v. infusion in 10 min. The placebo consisted of 100 ml dextrose 5%. The starting dose of monoHER was 100 mg/m(2). Dose escalation by 100% of the preceding dose took place after finishing each dose level until the protecting pharmacokinetic values for C (max) and AUC(infinity) (as observed in mice after 500 mg/kg monoHER i.p.) were reached and/or serious side effects were observed. The dose was escalated up to 1,500 mg/m(2). The mean values of C (max) and AUC(infinity) were 360+/-69.3 microM and 6.8+/-2.1 micromol min/ml, respectively. These values were comparable to the C (max) and AUC(infinity) observed under the protecting conditions in mice. No serious side effects occurred during the entire study. Thus, 1,500 mg/m(2) is a feasible and safe dose to be evaluated in a phase II study to investigate the protective properties of monoHER against doxorubicin-induced cardiotoxicity in cancer patients.


Optimizing subcutaneous injection of the gonadotropin-releasing hormone receptor antagonist degarelix.
The gonadotropin-releasing hormone (GnRH) receptor antagonist degarelix has several unique characteristics compared to luteinizing hormone-releasing hormone (LHRH) analogs used in the management of prostate cancer. Notable differences of GnRH receptor antagonists include no flare reaction, and a more rapid suppression of testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and prostate-specific antigen (PSA) compared to LHRH analogs. Despite emerging evidence supporting the use of GnRH receptor antagonists over the more widely used LHRH analogs in the management of prostate cancer, physicians may be reluctant to prescribe degarelix. They may be concerned about patient complaints about injection-site reactions (ISRs). The subcutaneous injection of degarelix has been associated with a higher rate of ISRs compared with the intramuscular injections of LHRH analogs. This "How I Do It" article describes techniques and strategies that have been developed by physicians and nurses to reduce the discomfort associated with the subcutaneous delivery of degarelix.


Direct purification and immobilization of his-tagged enzymes using unmodified nickel ferrite NiFe2O4 magnetic nanoparticles.
Purification of valuable engineered proteins and enzymes can be laborious, costly, and generating large amount of chemical waste. Whilst enzyme immobilization can enhance recycling and reuse of enzymes, conventional methods for immobilizing engineered enzymes from purified samples are also inefficient with multiple-step protocols, regarding both the carrier preparation and enzyme binding. Nickel ferrite magnetic nanoparticles (NiFe2O4 MNPs) offer distinct advantages in both purification and immobilization of enzymes. In this work, we demonstrate the preparation of NiFe2O4 MNPs via a one-step solvothermal synthesis and their use in direct enzyme binding from cell lysates. These NiFe2O4 MNPs have showed an average diameter of 8.9 ± 1.7 nm from TEM analysis and a magnetization at saturation (Ms) value of 53.0 emu g-1 from SQUID measurement. The nickel binding sites of the MNP surface allow direct binding of three his-tagged enzymes, D-phenylglycine aminotransferase (D-PhgAT), Halomonas elongata ω-transaminase (HeωT), and glucose dehydrogenase from Bacillus subtilis (BsGDH). It was found that the enzymatic activities of all immobilized samples directly prepared from cell lysates were comparable to those prepared from the conventional immobilization method using purified enzymes. Remarkably, D-PhgAT supported on NiFe2O4 MNPs also showed similar activity to the purified free enzyme. By comparing on both carrier preparation and enzyme immobilization protocols, use of NiFe2O4 MNPs for direct enzyme immobilization from cell lysate can significantly reduce the number of steps, time, and use of chemicals. Therefore, NiFe2O4 MNPs can offer considerable advantages for use in both enzyme immobilization and protein purification in pharmaceutical and other chemical industries.


[Choroidal type aneurysmal malformation of the vein of Galen associated with Dandy-Walker malformation in an adult].
Lasjaunias differentiated true vein of Galen aneurysmal malformations (MAVG), which drain to the median prosencephalic vein of Markowski, the embryonic precursor of the vein of Galen, from the aneurysmal dilatations of vein of Galen (VGDA), which reveal venous drainage into a dilated vein of Galen, but already formed. In angioarchitectural terms, MAVG can still be divided in two subtypes: the mural form and the choroidal form, the most common. The authors describe the clinical case of a 18 years-old female, without symptoms until July 2001, who presented an episode of impaired equilibrium associated with vomits and non-specific vision disturbances. This episode had an approximate duration of 6-8 hours with complete recovery. Neuroimaging studies were performed including CT, MRI and digital angiography, disclosed an aneurysmal vein of Galen malformation of choroidal type and Dandy-Walker malformation, association that the authors couldn't found so far described in the literature.


Novel coronavirus pandemic: A global health threat.
The world is facing one of its worst public health crises in modern history. Coronavirus 2019 (COVID-19) has shown how fragile our global preparedness for infectious diseases is. The world is a small-connected globe with short travel time between its remote parts. COVID-19 has spread globally and swiftly with major impacts on health, economy, and quality of life of communities. At this point in the time, April 9, 2020, >1,500,000 patients have been infected and >88,000 patients have died worldwide within the last 3 months. The status is evolving and the costly lessons learned over time are increasing. These lessons are global as this virus is. They involve different domains of health sciences including virology, public health, clinical, critical care, and disaster management. This review addresses our current knowledge of COVID-19 pandemic from the basic virology and transmission, through prevention, infection control, clinical management, and finally disaster management including the recovery period. This review has a multidisciplinary approach, which is needed at this time. After this difficult period passes, we have to carry the lessons we learned for the future so that we can be better prepared. One thing that has clearly emerged from this ongoing crisis is that infectious diseases have no borders and we have to work together, using the one world, one health approach, if we are to minimize the enormous impact such pandemics can cause.


A POU protein regulates mesodermal competence to FGF in Xenopus.
XLPOU91, a POU-homeobox gene is expressed in a narrow window during early Xenopus development. We show that ectopic expression of XLPOU91 RNA causes severe posterior truncations in embryos without inhibiting the formation of Spemann's organizer. Ectopic XLPOU91 expression also inhibits mesoderm induction by fibroblast growth factor (FGF) and activin in animal cap explants. Using antisense RNA, we depleted endogenous XLPOU91 protein in animal caps. Gastrula-stage animal caps expressing XLPOU91 antisense RNA do not lose competence to FGF, unlike controls, these animal caps express XBra after FGF treatment. Endogenous XLPOU91 levels are peaking when FGF mesoderm-inducing competence is lost in animal caps. Thus XLPOU91 protein may act as a competence switch during early development, as XLPOU91 levels increase in the embryo, the mesoderm response to FGF is lost.


Configurations of a two-tiered amplified gene expression system in adenoviral vectors designed to improve the specificity of in vivo prostate cancer imaging.
Effective treatment for recurrent, disseminated prostate cancer is notably limited. We have developed adenoviral vectors with a prostate-specific two-step transcriptional amplification (TSTA) system that would express therapeutic genes at a robust level to target metastatic disease. The TSTA system employs the prostate-specific antigen (PSA) promoter/enhancer to drive a potent synthetic activator, which in turn activates the expression of the therapeutic gene. In this study, we explored different configurations of this bipartite system and discovered that physical separation of the two TSTA components into E1 and E3 regions of adenovirus was able to enhance androgen regulation and cell-discriminatory expression. The TSTA vectors that express imaging reporter genes were assessed by noninvasive imaging technologies in animal models. The improved selectivity of the E1E3 configured vector was reflected in silenced ectopic expression in the lung. Significantly, the enhanced specificity of the E1E3 vector enabled the detection of lung metastasis of prostate cancer. An E1E3 TSTA vector that expresses the herpes simplex virus thymidine kinase gene can effectively direct positron emission tomography (PET) imaging of the tumor. The prostate-targeted gene delivery vectors with robust and cell-specific expression capability will advance the development of safe and effective imaging guided therapy for recurrent metastatic stages of prostate cancer.


Aciclovir protects against quinolinic-acid-induced oxidative neurotoxicity.
AIDS-related encephalopathy, including AIDS dementia complex (ADC) and the opportunistic disease, herpes simplex encephalitis (HSE), are postulated to arise due to the release of neurotoxic products, such as quinolinic acid (QUIN), by activated microglial cells in the brain. QUIN causes a cascade of events to occur, which leads to the production of reactive oxygen species (ROS), these being ultimately responsible for oxidative neurotoxicity. The antiherpes antiviral aciclovir has been reported to protect against neuron loss in HSE, but the mechanism for this neuroprotection is unknown. Therefore, this study was conducted to investigate whether aciclovir has the ability to inhibit QUIN-induced lipid peroxidation in rat brain homogenates, after in-vitro and in-vivo exposure to QUIN and aciclovir. The thiobarbituric acid (TBA) assay was the method used to analyse lipid peroxidation. Rat brains were also examined histologically after in-vivo exposure to visually assess whether neuron loss was suppressed. The results show that aciclovir inhibits the QUIN-induced lipid peroxidation, in a dose-dependent manner. Furthermore, aciclovir reduced necrosis of hippocampal neurons and retained the characteristic morphology, integrity and arrangement of these cells. Thus, it appears that aciclovir has neuroprotective properties, which could possibly be useful in the treatment of AIDS-related encephalopathy.


A Deletion Variant of the α2b-Adrenoceptor Modulates the Stress-Induced Shift from "Cognitive" to "Habit" Memory.
Stress induces a shift from hippocampus-based "cognitive" toward dorsal striatum-based "habitual" learning and memory. This shift is thought to have important implications for stress-related psychopathologies, including post-traumatic stress disorder (PTSD). However, there is large individual variability in the stress-induced bias toward habit memory, and the factors underlying this variability are completely unknown. Here we hypothesized that a functional deletion variant of the gene encoding the α2b-adrenoceptor (ADRA2B), which has been linked to emotional memory processes and increased PTSD risk, modulates the stress-induced shift from cognitive toward habit memory. In two independent experimental studies, healthy humans were genotyped for the ADRA2B deletion variant. After a stress or control manipulation, participants completed a dual-solution learning task while electroencephalographic (Study I) or fMRI measurements (Study II) were taken. Carriers compared with noncarriers of the ADRA2B deletion variant exhibited a significantly reduced bias toward habit memory after stress. fMRI results indicated that, whereas noncarriers of the ADRA2B deletion variant showed increased functional connectivity between amygdala and putamen after stress, this increase in connectivity was absent in carriers of the deletion variant, who instead showed overall enhanced connectivity between amygdala and entorhinal cortex. Our results indicate that a common genetic variation of the noradrenergic system modulates the impact of stress on the balance between cognitive and habitual memory systems, most likely via altered amygdala orchestration of these systems.SIGNIFICANCE STATEMENT Stressful events have a powerful effect on human learning and memory. Specifically, accumulating evidence suggests that stress favors more rigid dorsal striatum-dependent habit memory, at the expense of flexible hippocampus-dependent cognitive memory. Although this shift may have important implications for understanding mental disorders, such as post-traumatic stress disorder, little is known about the source of individual differences in the sensitivity for the stress-induced bias toward habit memory. We report here that a common genetic variation of the noradrenergic system, a known risk factor for post-traumatic stress disorder, modulates the stress-induced shift from cognitive to habit memory, most likely through altered crosstalk between the hippocampus and dorsal striatum with the amygdala, a key structure in emotional memory.


Double-blind comparison between doxapram and pethidine in the treatment of postanaesthetic shivering.
Sixty patients who shivered after routine surgery under general anaesthesia were allocated randomly to receive normal saline (n = 20), doxapram 1.5 mg kg-1 (n = 20) or pethidine 0.33 mg kg-1 (n = 20). Both doxapram and pethidine were effective in treating postoperative shivering 2-3 min after i.v. administration. In the group who received normal saline, 15 patients were still shivering 10 min after treatment, whilst in the doxapram group only three patients were shivering at that time. In the pethidine group, all patients had stopped shivering by 7 min after treatment. We conclude that both doxapram and pethidine were effective in the treatment of postoperative shivering.


The multicomponent reactions and their libraries for natural and preparative chemistry.
It was recently recognized that three different types of multi-component reactions (MCRs) exist. In preparative chemistry, the MCRs of type II form their products particularly efficiently. These reactions correspond to equilibria of educts and intermediate products, whose final products are formed practically irreversibly. In recent years, the four component reaction of the isocyanides (U-4CR) of type II and their unions with various reactions and MCRs have become an important industrial process for preparing products and their libraries. It has been demonstrated that all conceivable collections of U-4CR educts can be converted into the corresponding products. In the usual chemical reactions, only the substituents of the products can be varied, whereas the U-4CR and related reactions can also produce skeletally different types of products with diverse substituents. The preparative advantages of forming products by the one-pot MCRs and the great variety of the possible products are illustrated in this review.


Public views towards community health and hospital-based outpatient services and their utilisation in Zhejiang, China: a mixed methods study.
China is engaged in promoting community health services (CHS) nationwide. This study examines the public's views towards CHS and their utilisation of community-based and hospital-based outpatient services. A mixed methods study using qualitative interviews and a cross-sectional survey. The study was conducted among the public between September 2014 and September 2015 in Zhejiang province, China. Six focus groups and 13 individuals were interviewed. The questionnaire was completed by 1248 respondents (response rate: 83%). Utilisation of community-based and hospital-based outpatient services. Functions of CHS perceived by the public included provision of minor illness management, coordination, drug dispensing, follow-up care and patient education. However, many also showed a distrust in primary care providers' (PCPs) competence for confirming the initial diagnosis and management plan. As coordinators, PCPs' integrity was challenged, and PCPs were thought to be potential 'tuo er' (cunning agents who tried to lead patients to some notorious hospitals to make money). Survey results showed that 800 (64.1%) respondents visited hospital-based clinics and 688 (55.1%) visited CHS at least once in the past year. Compared with the uninsured group, those covered by Urban Resident Medical Insurance (adjusted OR (AOR)=1.95, 95% CI 1.24 to 3.07) and Urban Employee Medical Insurance (AOR=2.59, 95% CI 1.59 to 4.24) were more likely to use hospital-based services. Respondents who had a chronic condition were more likely than their counterparts to use both hospital-based services (AOR=1.72, 95% CI 1.18 to 2.49) and CHS (AOR=1.66, 95% CI 1.19 to 2.32). Income levels were positively associated with the likelihood of visiting hospital-based clinics (AOR=1.67, 95% CI 1.15 to 2.42) but negatively associated with the likelihood of using CHS (AOR=0.68, 95% CI 0.48 to 0.96). Demand of hospital-based outpatient services is much higher than the community-based outpatient services. Policy reformers need to take further actions to address the public distrust in PCPs to facilitate their gatekeeping role.


Internal Dynamics of Ionic Liquids over a Broad Temperature Range-The Role of the Cation Structure.
1H and 19F spin-lattice relaxation experiments have been performed for a series of ionic liquids sharing the same anion: bis(trifluoromethanesulfonyl)imide but including cations of different alkyl chain lengths: butyltriethylammonium, triethyloctylammonium, dodecyltriethylammo-nium and hexadecyltriethylammonium. The studies have been carried out in the temperature range from 383 to 108 K at the resonance frequency of 200 MHz (for 1H). A quantitative analysis of the relaxation data has revealed two dynamical processes for both kinds of ions. The dynamics have been successfully modeled in terms of the Arrhenius law. The timescales of the dynamical processes and their temperature evolution have been discussed in detail, depending on the structure of the cation.


Molecular epidemiology of outbreaks of gastroenteritis associated with small round-structured viruses in East Anglia, United Kingdom, during the 1996-1997 season.
During the winter season from November 1996 to May 1997, 550 fecal specimens were submitted from 94 outbreaks of gastroenteritis occurring in East Anglia, United Kingdom. These specimens were tested for the presence of small round-structured viruses (SRSVs) by electron microscopy, reverse transcriptase PCR, or both methods. SRSVs were shown to be associated with 64 of 94 (68%) of these outbreaks, of which 16 (25%) outbreaks occurred at a single location (Southend) within the region. Twenty-four specimens from 13 of the 16 SRSV-positive outbreaks occurring in Southend were available for genomic analysis, in which divergence within the RNA polymerase region of the SRSV genome was investigated. A further 27 specimens from 17 other SRSV-associated outbreaks, occurring at different locations within East Anglia but at the same time as those at Southend, were also studied. Fifty of the total of 51 (98%) specimens studied were shown to belong to genogroup II, and within this genogroup, 49 of 50 (98%) specimens were shown to be Grimsby-like viruses, with only one Mexico-like strain. Furthermore, phylogenetic analysis of the Grimsby-like viruses indicated clusterings according to the geographical location of the outbreak. One specimen contained a virus belonging to genogroup I, and this had the greatest sequence identity (83%) with Southampton virus.


Fabrication of sac-like hydrogel membranes for replicating curved tissue barriers on chips.
Current organ-on-a-chip (OOC) systems cannot mimic in vivo tissue barriers that feature curved geometries and rhythmic movement. This is due to the lack of a relevant membrane that can reproduce the natural biochemical and physical properties of a basement membrane, especially the characteristic sac-like structure possessed by multiple tissue barriers. To address this challenge, a sac-like hydrogel membrane is fabricated here using a one-step simple methodology inspired by soap bubble formation. Di-acrylated Pluronic® F127 (F127-DA) is a hydrogel that exhibits excellent mechanical properties, stably withstanding rhythmic mechanical stretching and fluid flow for at least 24 h. Using this hydrogel to make a membrane, a complex lung-on-a-chip device is successfully constructed, effectively replicating the alveolar-capillary barrier and demonstrating cellular function under physiological respiratory conditions. This membrane offers a crucial platform for replicating sac-like tissue barriers.


Community palliative care nurse experiences and perceptions of follow-up bereavement support visits to carers.
Community (district) nurses (CNs) are well positioned to provide follow-up home visits to bereaved families and carers of their recently deceased palliative clients. An Australian survey of CN's (n = 58, response rate 29%) described their experiences of bereavement support visits, perceptions of their role in bereavement care and their professional support needs. Although positive experiences were commonly reported, with 95% of participants considering bereavement follow-up visits as consistent with their role, 53% found the visits difficult for reasons such as the nurse or client not understanding the purpose, the CN's excessive personal identification with the client's situation, the emotional intensity of visits, and lack of confidence or skills despite prior training. The nature and quality of the CN's prior relationship with the bereaved family was an important determinant of the visits' success. Results highlight the value of bereavement support visits, while identifying professional development needs. Managing emotionally intense episodes should receive priority in preparing CN's for this challenging role.


Basilar-membrane responses to broadband noise modeled using linear filters with rational transfer functions.
Basilar-membrane responses to white Gaussian noise were recorded using laser velocimetry at basal sites of the chinchilla cochlea with characteristic frequencies near 10 kHz and first-order Wiener kernels were computed by cross correlation of the stimuli and the responses. The presence or absence of minimum-phase behavior was explored by fitting the kernels with discrete linear filters with rational transfer functions. Excellent fits to the kernels were obtained with filters with transfer functions including zeroes located outside the unit circle, implying nonminimum-phase behavior. These filters accurately predicted basilar-membrane responses to other noise stimuli presented at the same level as the stimulus for the kernel computation. Fits with all-pole and other minimum-phase discrete filters were inferior to fits with nonminimum-phase filters. Minimum-phase functions predicted from the amplitude functions of the Wiener kernels by Hilbert transforms were different from the measured phase curves. These results, which suggest that basilar-membrane responses do not have the minimum-phase property, challenge the validity of models of cochlear processing, which incorporate minimum-phase behavior.


[Determination of trace mercury in rice by microwave digestion-hydride generation-atomic fluorescence spectrometry].
It was reported in this paper that trace mercury in rice was determined by microwave digestion-hydride generation-atomic fluorescence spectrometry. The microwave digestion of samples was used, and the optimum conditions for the digestion were developed. The relative factors of hydride generation atomic fluorescence spectrometry for the determination were tested and discussed. The temperature and the relative humidity were discussed too. The detection limit was 0.005 ng x mL(-1). Mercury in reference materials (rice flour, GBW 08508) was determined also by the described method. The result obtained was in good agreement with standard value. The relative standard deviation (RSD) was 2.1%, and the recovery of mercury in rice samples was 95.2%-106.4%.


[The interaction between cells and fibrous dusts (author's transl)].
The shape of fibrous dust particles is an important factor for the biological effects in cell cultures. The fibres elicit a special mechanism of incorporation which is dependent on their length and diameter but independent of their chemical composition. Fibres with a length less than 5 mum are phagocytized rapidly and completely by the cells, comparable to granular dust particles or milled fibres. Phagocytosis of long inorganic fibers is delayed and/or remains incomplete. Incomplete incorporation of the fiber causes a localized discontinuity in the cell membrane. There results continuous liberation of intra-cellular enzymes, which is compensated by an increasing glycolytic metabolism. The interaction between cell and fibre means a chronic irritation which is discussed as a factor in tumour induction. Fibrous dusts, e.g. asbestos and glass fibers, induce the formation of polykaryotic giant cells by way of fusion; asbestos possibly also causes the interspecific fusion of cells. There are indications that in the process of asbestos-induced cell fusion integrated virus genomes are activated and infectious viruses are released. There seems to be a causal relationship between the fibrogenic and carcinogenic effects of inorganic dusts and their shape, i.e. their length and diameter, regardless of their chemical composition. Thus the fibrogenic and carcinogenic effect is limited by a minimal length and a maximal diameter. The findings are discussed with regard to the air burden of fibrous dusts and their implications for human health.


Increased hepatic insulin sensitivity together with decreased hepatic triglyceride stores in hormone-sensitive lipase-deficient mice.
Hormone-sensitive lipase (HSL) is a major enzyme for triglyceride (TG) lipolysis in adipose tissue. In HSL-knockout mice, plasma free fatty acid and TG levels are low, associated with low liver TG content. Because a decreased hepatic insulin sensitivity has been reported to be associated with high liver TG levels, our aim was to determine whether a hepatic TG content lower than normal, as observed in HSL-knockout mice, leads to increased hepatic insulin sensitivity. Therefore, hyperinsulinemic clamp experiments in combination with D-(3)H-glucose were used. Furthermore, hepatic insulin receptor and phosphorylated protein kinase B (PKB-P)/akt were analyzed by Western blotting. No significant differences where observed in insulin-mediated whole-body glucose uptake between HSL-knockout and control mice. Interestingly, hepatic insulin sensitivity of HSL-knockout mice was increased, because insulin caused a greater reduction in endogenous glucose production ( approximately 71% compared with approximately 31% in control mice; P < 0.05), despite decreased plasma adiponectin levels. PKB/akt phosphorylation and phosphatidylinositol-3-kinase activity was significantly higher in livers of HSL-knockout mice after insulin stimulation. In HSL-knockout mice, reduced hepatic TG stores result in an increased suppressive effect of insulin on hepatic glucose production, in line with an increased hepatic PKB-P/akt and phosphatidylinositol-3 kinase activity. Thus, hepatic insulin sensitivity is indeed increased after reducing hepatic TG stores below normal.


Meta-analysis: total parenteral nutrition versus total enteral nutrition in predicted severe acute pancreatitis.
Total parenteral nutrition (TPN) as a traditional mode of treatment in severe acute pancreatitis was still used widely in clinical work. In addition, enteral nutrition treatment methods have developed; early enteral nutrition has already been highlighted for severe acute pancreatitis, but the therapeutic risks versus benefits need to be studied. To compare total parenteral nutrition with total enteral nutrition (TEN) in patients with severe acute pancreatitis by performing a meta-analysis. Electronic databases including PubMed, EMBASE, Science Citation Index, were searched to find relevant randomized controlled trials. Two reviewers independently identified relevant trials evaluating the effect of total parenteral nutrition and early enteral nutrion. Outcome measures were the mortality, hospital length of stay, infectious complications, duration of nutrition, organ failure and surgical intervention. Eight randomized controlled trials (RCTs) including 381 patients were identified. Meta-analysis demonstrated that TEN was significantly superior to TPN when considering mortality [p=0.001, 95%CI 0.37(0.21-0.68)], infectious complications [p=0.004, 95%CI 0.46(0.27-0.78)], organ failure [p=0.02, 95%CI 0.44(0.22-0.88)] and surgical intervention [p=0.003, 95%CI 0.41(0.23-0.74)].While no difference between TEN and TPN when considering the hospital length of stay [p=0.22, 95%CI -14.10(-36.48-8.26)] and as for duration of nutrition [p=0.72, 95%CI -1.50(-9.56-6.56)] there was not enough data to compare the differences. Total enteral nutritional support is associated with lower mortality, fewer infectious complications, decreased organ failure and surgical intervention rate compared to parenteral nutritional support.


Depressive disorders in childhood. I. A longitudinal prospective study of characteristics and recovery.
In the context of a prospective, longitudinal, and controlled nosologic study, the characteristics and diagnostic validity of major depressive disorder, dysthymic disorder, and adjustment disorder with depressed mood were examined in a school-aged cohort. The entities were distinguishable on several dimensions such as age at onset and pattern of recovery. Time to recovery from onset was most favorable for the adjustment disorders (90% remission rate during nine months) and least so for the dysthymias (89% remission rate during six years). Major depression and dysthymia were similar with respect to the prevalence of concurrent nonaffective disorders. For both, early age at onset predicted a more protracted illness. Treatment contacts, none of which were under the control of the investigators, had no clear impact on recovery from the depressions.


Medullary inspiratory activity during opossum development.
Discharge properties of medullary neurons associated with inspiration were assessed during opossum development. Location of these neurons and projections to the spinal cord were also evaluated. Studies were performed on suckling opossums, from 15-20 days of age to weaning (approximately 100 days), as well as adult animals, anesthetized with Inactin. Most cells either discharged during inspiration (I cells) or were phase-spanning between late expiration and inspiration (EI cells). Number of spikes per breath for I and EI cells significantly increased as the animal matured. For younger opossums (up to approximately 60 days of age), bulbospinal I and EI cells discharged an average of 1.9 spikes per breath, with a minimum interspike interval of 59 ms. Their conduction velocities, determined by use of antidromic stimulation, were in the unmyelinated range (less than 2 m/s). The occurrence of few spikes per breath with long interspike interval, in bulbospinal cells of the younger opossums, suggests that inspiration is not subject to fine feedback control and that rate of high-frequency motor bursts is relatively slow.


Decreased Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel 2 Activity in a Rat Model of Absence Epilepsy and the Effect of ZD7288, an Ih Inhibitor, on the Spike-and-Wave Discharges.
Hyperpolarization-activated cyclic nucleotide-gated (HCN) channel currents of Ih and absence epilepsy seizures are associated, but studies reveal differential results. In our study, we aimed to investigate the role of the HCN channels on the expression of spike-and-wave discharges (SWDs) using the Genetic Absence Epilepsy Rats from Strasbourg (GAERS) model. HCN isoform levels from isolated brains of both naïve nonepileptic Wistar and GAERS groups were evaluated by enzyme-linked immunosorbent assay. ZD7288, an Ih inhibitor as well as an HCN channel antagonist, was administered intracerebroventricularly to the adult GAERS groups, and to evaluate their SWD activities, electroencephalography was recorded. The effect of ZD7288 on the cumulative total duration and number of SWDs and the mean duration of each SWD complex was evaluated. The HCN2 levels in the cortex and hippocampus of the GAERS group were lower compared to the naïve nonepileptic Wistar group (p < 0.05). ZD7288 increased the number of SWDs at the 20th and 120th min with the highest administered dose of 7 μg (p < 0.05). The Ih inhibitor ZD7288 increased the number of SWDs in a genetic absence epilepsy rat model, although this increase may not be significant due to the inconsistent time-dependent effects. In GAERS, the cortical and hippocampal HCN2 channel levels were significantly lower compared to the control group. Further studies are needed with higher doses of ZD7288 to determine if the effects will increase drastically.


Risk of secondary malignancy (including breast) in patients with mismatch-repair protein deficiency.
Lynch syndrome (LS) is an autosomal dominant inherited disease that is associated with an increased risk for colorectal and endometrial cancer due to germline mutations in mismatch-repair (MMR) genes. Whereas primary tumors in this syndrome are widely recognized, the relative risk(s) of secondary malignancies, particularly breast cancer, in LS patients are still poorly characterized. To provide an improved assessment of these risks, MMR status was evaluated in secondary tumors from a series of patients with index tumors of known MMR status (both proficient and deficient). A total of 1252 tumors (index tumors) and all secondary malignancies were tested for MMR by immunohistochemistry (MSH2, MSH6, MLH1, PMS2) between 1992 and 2013. Tumors with MLH1/PMS2 deficiency were tested for hypermethylation or BRAF mutation, when appropriate. Of the 1252 index tumors, 162 were MMR deficient (dMMR), and, of that subset, 32 secondary tumors were identified (19.7%). In contrast, 80 secondary tumors were identified in the proficient (intact) group (7.3%). Although secondary malignancies were more common in the dMMR group (P=0.0001), there was no trend in tumor type. Specifically, breast cancer was not overly represented in the dMMR group. When secondary tumors had dMMR, they were more likely to have deficiency in MSH2/MSH6 than in MLH1/PMS2 (P=0.01). Of the patients with tumors exhibiting dMMR, women were more likely to have a dMMR secondary tumor in this series (P=0.0001); however, breast cancer was not overly represented, and our study provides no evidence that it is more frequent in LS. MSH2/MSH6 deficiency is more commonly associated with a secondary tumor compared with MLH1/PMS2 deficiency, when methylation/BRAF status is taken into account.


[Human rabies transmitted by dogs: risk areas in Minas Gerais, Brazil, 1991-1999].
A retrospective study based on observation with the objective of identifying and characterizing the different risk areas for rabies transmission by dogs took place in Minas Gerais State, Brazil, from 1991 to 1999. Indicators confirmed occurrences of canine and feline rabies, notification of human rabies, and administration of appropriate vaccination. The Minas Gerais State Health System is divided into 25 Regional Health Centers, which are linked to the State Health Department (SES-MG). These Health Centers were utilized in the study. The results of 2,845 records of laboratory diagnosis for canine, feline, and human rabies were analyzed. Consolidated SES-MG reports from 1997 to 1999 for rabies vaccination and notification records for cases of human rabies from the National Health Foundation (FUNASA) were also used. In order to verify the local reality, a semi-structured interview with each regional program director was conducted. Minas Gerais presents four different risk modalities, classified as zero, low, medium, and high.


Low dose radiation mechanisms: The certainty of uncertainty.
This paper reviews the current understanding of low dose radiobiology, and how it has evolved from classical target theory. It highlights the uncertainty around low dose effects, which is due in part to the complexity of "context" surrounding the ultimate expression of biological effects following low dose exposure. The paper makes special reference to low dose non-targeted effects which, are currently ignored in radiation protection and population level risk assessment, because it is unclear what they mean for risk. The view of the authors is that this "lack of clarity" about what the effects mean is precisely the point. It indicates the uncertainty of outcomes after a given exposure. The uncertainty stems from multiple outcome options resulting from the intrinsic uncertainty of the stochastic interaction of low dose radiation with matter. This uncertainty should be embraced rather than eschewed. The impacts of the uncertainties identified in this paper is explored and an approach to quantifying mutation probability in relation to dose is presented.


Promotion of health by soy isoflavones: efficacy, benefit and safety concerns.
Cardiovascular diseases, osteoporosis-related hip fractures, and various cancers of the colon, prostate, uterus, and breast are remarkably less prevalent in Asia than in other industrialized countries. It is believed that the large consumption of soy products in Asian countries is contributory to the reduction of these chronic disorders. Genistein is a major isoflavone found in most soy products and plays an important role in the promotion of human health. Extensive epidemiological, in vitro, and animal studies have been performed, and most studies indicate that genistein has beneficial effects on a multitude of human disorders, including cancers, cardiovascular diseases, osteoporosis, and postmenopausal symptoms. To date, there is an abundance of promising studies supporting genistein's potential uses, but further research is still needed to validate its preventative and therapeutic efficacy. In addition, the adverse effects of genistein have drawn public attention. More studies are required to assess the potential detrimental effect of genistein, and a benefit-risk ratio should be considered before future clinical studies are performed.


Timely levodopa (LD) administration prolongs survival in Parkinson's disease.
Levodopa (LD) is the most effective drug for symptomatic control of Parkinsonism (PS). As prolonged LD therapy leads to response failure; is associated with adverse effects; may accelerate disease process, and its impact on life expectancy is controversial, delaying LD use has been recommended. We studied 934 PS cases including 859 Parkinson's disease (PD) patients over twenty-two years (1968-1990) to identify: (i) survival change coincident with widespread LD use and (ii) the critical level of disease severity when LD influences survival. Survival was measured from the date of first assessment until the last evaluation for a cumulative 3563 person-years of follow-up and compared with that expected in the general population of the same age, sex and year of birth. A Cox regression model was used to identify independent clinical predictors of survival. The observed to expected survival was reduced significantly in PS. Dementia and advanced disability at initial assessment and the final non-PD diagnosis, each was independently associated with poor survival. Survival improved significantly coincident with the widespread unrestricted access to LD (p = 0.024 - < 0.001) compared to pre-levodopa era cases. Life expectancy was longest in the more recent non-demented PD cases. When the major disease and treatment related factors were considered the timely use of levodopa in the recent cases resulted in improved survival compared to the earlier cases who received LD later in the course of illness. When LD naive cases were considered, survival was significantly reduced if the drug was delayed until Stage 3 Hoehn and Yahr, but not so if the patients remained untreated during Stages 1 and 2 Hoehn andYahr[1]. Life expectancy in PD has increased significantly since the widespread use of LD. This benefit is evident only if LD therapy is initiated before the patient reaches Stage 2.5 disability [2].


Indole-TEMPO conjugates alleviate ischemia-reperfusion injury via attenuation of oxidative stress and preservation of mitochondrial function.
Mitochondrial oxidative damage contributes to a wide range of pathologies including ischemia/reperfusion injury. Accordingly, protecting mitochondria from oxidative damage should possess therapeutic relevance. In the present study, we have designed and synthesized a series of novel indole-TEMPO conjugates that manifested good anti-inflammatory properties in a murine model of xylene-induced ear edema. We have demonstrated that these compounds can protect cells from simulated ischemia/reperfusion (s-I/R)-induced reactive oxygen species (ROS) overproduction and mitochondrial dysfunction. Furthermore, we have demonstrated that indole-TEMPO conjugates can attenuate organ damage induced in rodents via intestinal I/R injury. We therefore propose that the pharmacological profile and mechanism of action of these indole-TEMPO conjugates involve convergent roles, including the ability to decrease free radical production via lipid peroxidation which couples to an associated decrease in ROS-mediated activation of the inflammatory process. We further hypothesize that the protective effects of indole-TEMPO conjugates partially reside in maintaining optimal mitochondrial function.


Effects of resistance exercises on inhibitory control and plasma epinephrine levels: A registered report of a crossover randomized controlled trial.
According to the locus coeruleus-norepinephrine (LC-NE) theory, activity of the LC, the major releaser of NE in the brain, regulates inhibitory control. As there is reciprocal communication between circulating epinephrine and the LC, plasma epinephrine is used as the index of LC-NE activity. The aim of this crossover randomized controlled trial is to expand on previous findings by investigating the effects of free-weight, multiple-joint, and structural barbell resistance exercises. Previous studies have had some methodological limitations, such as failure to report the process of randomization, absence of resistance exercise familiarization before the maximal strength testing, and lack of protocol publication. To address these issues, this study incorporates resistance exercise familiarization, transparent reporting of randomization, and submission as a registered report. The results suggest that a single session of resistance exercise (barbell squat, press, and deadlift) with an intensity of 65%-78% 1RM for five repetitions (70%-90% relative intensity) and three sets with 3-min rest intervals improved Stroop congruent reaction time (RT) only (t(27) = -2.663, mean reduction = -15 ms, p = .013, 95% CI [-26, -3]). No significant enhancements were observed in Stroop incongruent RT, inhibitory control as indexed by Stroop effect, or inhibitory control as indexed by the RT difference between the Stroop task and the simple reaction task. Moreover, the alterations in plasma epinephrine levels did not significantly associate with changes in any measure of cognitive performance.


Direct Observation of Charge Collection at Nanometer-Scale Iodide-Rich Perovskites during Halide Exchange Reaction on CH3NH3PbBr3.
Organolead halide perovskites MAPbX3 (MA = CH3NH3+, X = Cl-, Br-, or I-) are known to undergo reversible halide exchange reactions, enabling bandgap tuning over the visible light region. Using single-particle photoluminescence (PL) imaging for in situ observation, we have studied the structure-dependent charge dynamics during halide exchange with iodide ions on an MAPbBr3 crystal. In particular, we optically detected nanometer-scale iodide-rich domains (i.e., MAPbBrI2) and found that their lifetimes of several tens of milliseconds are limited by reaction with diffusing vacancies. Furthermore, it was discovered that these domains effectively collect the charge carriers from the bulk crystal, thus resulting in amplified spontaneous emission (ASE) under continuous-wave laser irradiation. Our findings will provide direction for development of perovskite heterostructures with enhanced charge utilization.


Insulin and tri-iodothyronine induce glucokinase mRNA in primary cultures of neonatal rat hepatocytes.
Glucokinase (EC 126.96.36.199) first appears in the liver of the rat 2 weeks after birth and increases after weaning on to a high-carbohydrate diet. We investigated the hormonal regulation of glucokinase (GK) mRNA in primary cultures of hepatocytes from 10-12-day-old suckling rats. GK mRNA was undetectable in such cells after 48 h of culture in serum-free medium devoid of hormones. Addition of insulin or tri-iodothyronine (T3) to the medium resulted in induction of GK mRNA. The effects of insulin and T3 were dose-dependent and additive. Dexamethasone alone did not induce GK mRNA, but enhanced the response to insulin and decreased the response to T3. Induction of GK mRNA by insulin was not affected when the medium glucose concentration was varied between 5 and 15 mM, nor when culture was conducted in glucose-free medium supplemented with lactate and pyruvate or galactose. The time course of initial accumulation of GK mRNA in response to insulin was characterized by a lag of 12 h and an induction plateau reached after 36 h. If hepatocytes were then withdrawn from insulin for 24 h and subsequently subjected to a secondary stimulation by insulin, GK mRNA re-accumulated with much faster kinetics and reached the fully induced level within 8 h. Both primary and secondary responses to insulin were abolished by actinomycin D. These results provide insight into the role of hormonal stimuli in the ontogenic development of hepatic glucokinase.


Stefin B Inhibits NLRP3 Inflammasome Activation via AMPK/mTOR Signalling.
Stefin B (cystatin B) is an inhibitor of lysosomal and nuclear cysteine cathepsins. The gene for stefin B is located on human chromosome 21 and its expression is upregulated in the brains of individuals with Down syndrome. Biallelic loss-of-function mutations in the stefin B gene lead to Unverricht-Lundborg disease-progressive myoclonus epilepsy type 1 (EPM1) in humans. In our past study, we demonstrated that mice lacking stefin B were significantly more sensitive to sepsis induced by lipopolysaccharide (LPS) and secreted higher levels of interleukin 1-β (IL-1β) due to increased inflammasome activation in bone marrow-derived macrophages. Here, we report lower interleukin 1-β processing and caspase-11 expression in bone marrow-derived macrophages prepared from mice that have an additional copy of the stefin B gene. Increased expression of stefin B downregulated mitochondrial reactive oxygen species (ROS) generation and lowered the NLR family pyrin domain containing 3 (NLRP3) inflammasome activation in macrophages. We determined higher AMP-activated kinase phosphorylation and downregulation of mTOR activity in stefin B trisomic macrophages-macrophages with increased stefin B expression. Our study showed that increased stefin B expression downregulated mitochondrial ROS generation and increased autophagy. The present work contributes to a better understanding of the role of stefin B in regulation of autophagy and inflammasome activation in macrophages and could help to develop new treatments.


The validity of stimulus-evoked EMG for studying muscle fatigue characteristics of paraplegic subjects during dynamic cycling movement.
The fatigue characteristics of paralyzed muscles were investigated during dynamic cycling movement induced by functional electrical stimulation (FES). The peak-to-peak (PTP) amplitude of stimulus-evoked electromyogram (EMG), after suppression of stimulus artifact, was adopted as fatigue indicator. Compared to static contraction, the effects of dynamic movement factors on the stimulus-evoked EMG, such as the intermittent stimulation, joint angle, and contraction speed, were first evaluated in separate experiments. The results of isolated tests laid the foundation for interpreting the data obtained in two FES-cycling experiments, performed under maximum stimulation or in controlled cycling speeds. The effects of intermittent stimulation and joint angle caused periodic changes in PTP amplitude which can be alleviated by averaging the PTP amplitude of one cycle. Under the same stimulation intensity, our results indicated that slower muscle contraction speed would have larger PTP amplitude and vice versa. For the limited number of subjects with paraplegia studied, our results showed that the use of EMG PTP as reliable muscle fatigue indicator during dynamic movement is only valid at the same cycling speed or corresponding contraction speed. The decline of the PTP amplitude decreased with the decay of muscle force can be observed during cycling movement; however, reduction of cycling speed had the opposite effect on PTP amplitude. Observations from the hyperbolic modeling of fatigue process demonstrated that the EMG PTP of a fatigued muscle under dynamic movement decreased at a slower rate.


Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia.
The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSAmT/mG mice. We observe no evidence of short-term toxicity with a widespread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells.


Spontaneous oxygen radical generation by sickle erythrocytes.
Since the various membrane abnormalities of sickle erythrocytes might result from excessive accumulation of oxidant damage, we have measured the generation of superoxide, peroxide, and hydroxyl radical by normal and sickle erythrocytes using assays involving reduction of cytochrome c, aminotriazole inhibition of catalase, and methane evolution from dimethyl sulfoxide, respectively. Compared with normal erythrocytes, sickle erythrocytes spontaneously generate approximately twice as much superoxide, peroxide, and hydroxyl radical. One possible source of hydroxyl radical generation was identified as hemichrome, excessive amounts of which are bound to sickle erythrocyte membranes. Hemichrome did not generate hydroxyl radical when exposed to superoxide alone or peroxide alone. However, in the presence of both superoxide and peroxide, hemichrome greatly facilitated hydroxyl radical generation. Supporting this, normal erythrocyte membranes induced to acquire sickle hemichrome concomitantly acquired an enhanced ability to mediate hydroxyl radical generation. Finally, sickle erythrocyte membranes greatly enhanced superoxide/peroxide-driven hydroxyl radical generation as compared with normal erythrocyte membranes. These data suggest that an excessive accumulation of oxidant damage in sickle erythrocyte membranes might contribute to the accelerated membrane senescence of these cells. They further indicate that accumulation of oxidant damage could be a determinant of normal erythrocyte membrane senescence.


Normal and collapsed trachea in the dog: scanning electron microscopy study.
Scanning electron microscopy was used to examine canine tracheal cartilage and to determine the relationship between the fibrous and amorphous matrix in this tissue. Collapsed tracheal (CT) cartilage was hypocellular, compared with normal tracheal cartilage. The amorphous matrix of CT cartilage had a porous, fissured texture and did not have the homogeneous appearance of normal tracheal cartilage. Capillaries were seen to pass through CT cartilage. Randomly distributed connective tissue fibers, evident in CT cartilage matrix, were frequently attached to irregular shapes and sizes of amorphous matrix.


Molecular epidemiology and clinical characteristics of hepatitis B identified through the French mandatory notification system.
Strains responsible for acute hepatitis B infections (AHB) in France have not been characterized. This study was first designed to analyze the molecular epidemiology of AHB and second to describe the differences between AHB and chronic hepatitis B (CHB) exacerbations. This prospective study was based on the French mandatory notification system for AHB. 147 samples corresponding to declared cases were shipped to a central laboratory for classification as AHB or CHB according to the level of anti-HBc IgM and anti-HBc avidity. Based on biological marker values and file examination, 75 cases (59%) were classified as AHB. Independently of the acute or chronic status, genotype A (57%), D (22%) and E (14%) were the most prevalent and no phylogenetic clustering was observed among HBV sequences (n=68). Precore or basal core-promoter variants were not particularly associated with disease severity but were more prevalent in CHB. No antiviral resistant strains or immune-escape HBsAg was observed. HBV viral loads in AHB or CHB were comparable but with opposite distributions. ALT levels reached 10 times the upper normal value in 94% of AHB but only in 24% of CHB. After rigorous classification, no major difference at the genetic level was found between HBV strains isolated from AHB and CHB. Absence of potentially deleterious variant detection is reassuring. When based upon HBsAg and anti-HBc IgM determination, AHB notification may falsely include more than 40% CHB, leading to an important risk of bias in national surveillance programs of AHB.


A spectroscopic study of nicotine analogue 2-phenylpyrrolidine (PPD) using resonant two-photon ionization (R2PI), microwave, and 2D NMR techniques.
Conformational preferences of the nicotine analogue 2-phenylpyrrolidine (PPD) have been studied in both gaseous and solution phases. Theoretical calculations at the MP2 and B3LYP levels point to 5-6 stable conformers which differ in three degrees of conformational freedom; torsion between the two rings, inversion at the pyrrolidine (PY) amine, and PY ring puckering, characterized using the Cremer-Pople definition for pseudorotation. Only one conformer has a trans arrangement between the amino hydrogen and the phenyl substituent. It is 6-8 kJ mol(-1) more stable than the cis conformers, has a perpendicular ring arrangement, and puckers at the nitrogen atom--similar to structures reported for nicotine. Resonant two-photon ionization (R2PI) data, including hole burn spectra, indicate only one conformer is present in the free jet expansion, and band contour analysis suggests assignment to the trans conformer. Confirmation was provided by microwave spectroscopy. Fifty-seven lines measured in the 48-72 GHz region were assigned to 206 b-type transitions and fitted to yield rotational constants within 2 MHz of MP2 values predicted for the trans conformer. The solution-phase conformers of PPD were studied using 1D and 2D (1)H NMR spectroscopy and solvent-based theoretical calculations. In marked contrast to the gas phase, NMR data reveals only cis conformers present in solution. Calculations confirm increased stability for these conformers when placed in simulated chloroform or water environments. Solvent molecules are believed to disrupt a crucial N...H(ortho) stabilizing interaction present within the trans conformer.


Sexual and physical abuse history and adult sexual risk behaviors: relationships among women and potential mediators.
While research has supported associations between experiencing abuse and engaging in risky sexual behaviors during adolescence, research regarding these associations among adult women is much more equivocal. In addition, few studies have attempted to identify potential pathways from abuse experiences to sexual risk behaviors. The current study examined the associations between a history of physical or sexual abuse and recent sexual risk behaviors among adult women. Additionally, this study evaluated binge drinking and depressive symptomatology as potential mediators of any relationships between abuse history and sexual risk behaviors. A total of 1,428 women between 18 and 40 years of age attending family planning clinic appointments completed a self-report survey regarding their recent sexual behaviors and sexual and physical abuse history. Logistic regressions using backward elimination were conducted to identify factors associated with sexual risk behavior. A history of physical abuse by a romantic partner was associated with several sexual risk behaviors. Few significant associations between intrafamilial physical or sexual abuse and recent sexual risk behaviors were found. Additionally, there was no evidence that these relationships were mediated by binge drinking or depressive symptomatology. Familial abuse experiences are not necessarily associated with recent sexual risk behaviors among adults. In contrast, physical abuse experiences, particularly those perpetrated by a romantic partner, are associated with engaging in adult sexual risk behaviors among women. However, these associations are not mediated by alcohol use or depressive symptomatology.


Immune Axonal Polyneuropathy.
Immune axonal polyneuropathy is caused by a diverse group of disorders that share similar presentations and treatment regimens. This article focuses on the clinical findings, evaluation, and management of immune-mediated causes of axonal polyneuropathy, focusing primarily on large fiber sensorimotor polyneuropathy. Specific characteristics of an immune-mediated polyneuropathy have been incorporated in a new diagnostic screening tool that is highly sensitive and can easily be used in the outpatient clinic setting. New insights into autoantibodies may help identify the presence of an underlying autoimmune or paraneoplastic disease as the cause of a polyneuropathy. This article provides readers with further understanding into the autoimmune causes of axonal polyneuropathy and will help the clinician recognize key clinical features that may lead to timely diagnosis and treatment.


Influence of centriole behavior on the first spindle formation in zygotes of the brown alga Fucus distichus (Fucales, Phaeophyceae).
The influence of centrioles, derived from the sperm flagellar basal bodies, and the centrosomal material (MTOCs) on spindle formation in the brown alga Fucus distichus (oogamous) was studied by immunofluorescence microscopy using anti-centrin and anti-beta-tubulin antibodies. In contrast to a bipolar spindle, which is formed after normal fertilization, a multipolar spindle was formed in polyspermic zygote. The number of mitotic poles in polyspermic zygotes was double the number of sperm involved in fertilization. As an anti-centrin staining spot (centrioles) was located at these poles, the multipolar spindles in polyspermic zygotes were produced by the supplementary centrioles. When anucleate egg fragments were fertilized, chromosome condensation and mitosis did not occur in the sperm nucleus. Two anti-centrin staining spots could be detected, microtubules (MTs) radiated from nearby, but the mitotic spindle was never produced. When a single sperm fertilized multinucleate eggs (polygyny), abnormal spindles were also observed. In addition to two mitotic poles containing anti-centrin staining spots, extra mitotic poles without anti-centrin staining spots were also formed, and as a result multipolar spindles were formed. When karyogamy was blocked with colchicine, it became clear that the egg nucleus proceeded independently into mitosis accompanying chromosome condensation. A monoastral spindle could be frequently observed, and in rare cases a barrel-shaped spindle was formed. However, when a sperm nucleus was located near an egg nucleus, the two anti-centrin staining spots shifted to the egg nucleus from the sperm nucleus. In this case, a normal spindle was formed, the egg chromosomes arranged at the equator, and the associated MTs elongated from one pole of the egg spindle toward the sperm chromosomes which were scattered. From these results, it became clear that paternal centrioles derived from the sperm have a crucial role in spindle formation in the brown algae, such as they do during animal fertilization. However, paternal centrioles were not adequate for the functional centrosome during spindle formation. We speculated that centrosomal materials from the egg cytoplasm aggregate around the sperm centrioles and are needed for centrosomal activation.


A continuum method for determining membrane protein insertion energies and the problem of charged residues.
Continuum electrostatic approaches have been extremely successful at describing the charged nature of soluble proteins and how they interact with binding partners. However, it is unclear whether continuum methods can be used to quantitatively understand the energetics of membrane protein insertion and stability. Recent translation experiments suggest that the energy required to insert charged peptides into membranes is much smaller than predicted by present continuum theories. Atomistic simulations have pointed to bilayer inhomogeneity and membrane deformation around buried charged groups as two critical features that are neglected in simpler models. Here, we develop a fully continuum method that circumvents both of these shortcomings by using elasticity theory to determine the shape of the deformed membrane and then subsequently uses this shape to carry out continuum electrostatics calculations. Our method does an excellent job of quantitatively matching results from detailed molecular dynamics simulations at a tiny fraction of the computational cost. We expect that this method will be ideal for studying large membrane protein complexes.


Health Services Research Spending and Healthcare System Impact Comment on "Public Spending on Health Service and Policy Research in Canada, the United Kingdom, and the United States: A Modest Proposal".
The challenges associated with translating health services and policy research (HSPR) evidence into practice are many and long-standing. Indeed, those challenges have themselves spawned new areas of research, including knowledge translation and implementation science. These sub-disciplines have increased our understanding of the critical success factors associated with the uptake of research evidence into (system) practice. Engaging those for whom research evidence is likely to help solve implementation and/or policy problems, and ensuring that they are key partners throughout the research life-cycle, appear to us (based on current evidence) to be the most direct and effective paths to improved knowledge translation. In that regard, building on Canada's recent Strategy for Patient Oriented Research (SPOR) would seem to offer considerable promise. The "modest" proposals offered by Thakkar and Sullivan seem less likely to bear fruit.


Outcomes after surgical resection of head and neck paragangliomas: a review of 61 patients.
We reviewed the postoperative functional outcome following surgical resection of paragangliomas in patients with and without preoperative cranial nerve dysfunction. Patients who underwent surgical resections of head and neck paragangliomas were reviewed with functional outcomes defined as feeding tube and/or tracheostomy dependence, need for vocal cord medialization, and incidence of cerebral vascular accidents as primary end points. Secondary end points included pre- and postoperative function of lower cranial nerves and the impact of this dysfunction on long-term functional status. Sixty-one patients were identified: 27 with carotid paraganglioma (CP), 21 with jugular paraganglioma (JP), 8 with tympanic paragangliomas, 4 with vagal paragangliomas (VPs), and 1 with aortopulmonary paraganglioma. Following resection, 8 patients were feeding tube dependent, 14 patients required vocal cord medialization, 2 patients suffered strokes, but no patients required tracheostomy tubes. Twenty percent of patients (4/20) with JP and postoperative cranial neuropathies were feeding tube dependent, and 80% of patients (4/5) with CP and postoperative cranial nerve dysfunction were feeding tube dependent. Cranial nerve deficits were more common in patients with JP relative to those with CP. However, when cranial nerve dysfunction was present, our patients with CP had a higher incidence of temporary feeding tube dependence. Overall, 98% of patients were able to resume oral nutrition.


The Cox model is better than the Fine and Gray model when estimating relative revision risks from arthroplasty register data.
Background and purpose - Analysis of the revision-free survival of knee and hip prostheses has traditionally been performed using Kaplan-Meier analysis and Cox regression. The competing risk problem that is related to patients who die during follow-up has recently been increasingly discussed, not least with regard to the problem of choosing a suitable statistical method for the analysis. We compared the results from analyses of Cox models and Fine and Gray models. Methods - We used data simulation based on parameter estimates from the Swedish Knee Arthroplasty Register and assessed hypothetical effects of the studied risk factors. Results - The Cox model provided more adequate results. Interpretation - The parameter estimates from the Fine and Gray model can be misleading if interpreted in terms of relative risk.


Radiofrequency ablation of large renal angiomyolipoma: median-term follow-up.
To study the feasibility of percutaneous radiofrequency ablation (RFA) of large angiomyolipomas (AMLs) using saline-cooled electrodes. Institutional Review Board approval for the study was received. Four patients (all female, age range 33-67 years) with large AMLs (maximal axis 6.1-32.4 cm) not suitable for embolotherapy or surgery consented to a trial of RFA. Procedures were performed under computerized tomographic guidance using 14G saline-infused electrodes. Two ablations (diameter 4-7 cm) were undertaken in each patient. Variables studied were technical success, treatment safety, alteration of tumor consistency, tumor size, effect on renal function, and medium-term freedom from haemorrhage. All four patients underwent successful RFA without any intraprocedural complications. There has been no haemorrhage, or new renal specific symptom, during a minimum 48-month period, and normal renal function has been normal. On follow-up radiological imaging, the tumors have become fattier with involution of the soft-tissue elements (soft tissue-to-total tumor ratio decreased mean [range] of 0.26 [0.14-0.48] to 0.17 [0.04-0.34] U; p = 0.04 [paired Student t test]). Further evidence of treatment effect was the development of a capsule around the ablation zone, but there was no change in overall tumor volume (mean [range] 1,120 [118-2,845] to 1150 [90-3,013] ml; p = 1 [paired Student t test]). RFA of large AMLs is technically feasible using saline-infused electrodes. The soft-tissue elements decreased in volume; the tumors become fattier; and there has been no renal haemorrhage during a 48-month period.


Antibacterial Effect of Eicosapentaenoic Acid against Bacillus cereus and Staphylococcus aureus: Killing Kinetics, Selection for Resistance, and Potential Cellular Target.
Polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA; C20:5n-3), are attracting interest as possible new topical antibacterial agents, particularly due to their potency and perceived safety. However, relatively little is known of the underlying mechanism of antibacterial action of EPA or whether bacteria can develop resistance quickly against this or similar compounds. Therefore, the aim of this present study was to determine the mechanism of antibacterial action of EPA and investigate whether bacteria could develop reduced susceptibility to this fatty acid upon repeated exposure. Against two common Gram-positive human pathogens, Bacillus cereus and Staphylococcus aureus, EPA inhibited bacterial growth with a minimum inhibitory concentration of 64 mg/L, while minimum bactericidal concentrations were 64 mg/L and 128 mg/L for B. cereus and S. aureus, respectively. Both species were killed completely in EPA at 128 mg/L within 15 min at 37 °C, while reduced bacterial viability was associated with increased release of 260-nm-absorbing material from the bacterial cells. Taken together, these observations suggest that EPA likely kills B. cereus and S. aureus by disrupting the cell membrane, ultimately leading to cell lysis. Serial passage of the strains in the presence of sub-inhibitory concentrations of EPA did not lead to the emergence or selection of strains with reduced susceptibility to EPA during 13 passages. This present study provides data that may support the development of EPA and other fatty acids as antibacterial agents for cosmetic and pharmaceutical applications.


Experimental infection of lactating goats with Leptospira interrogans serovars pomona and hardjo.
Pathogenesis of Leptospira interrogans serovars pomona and hardjo was evaluated in 14 lactating goats. Although mild clinical signs of leptospiral infection characterized by pyrexia and reduction in milk yield appeared in some animals, a consistent clinical pattern was not observed in the inoculated animals. The pomona serovar was isolated from the kidney of 1 of the 4 goats inoculated with serovar pomona. The hardjo serovar (strain UI 750) was isolated in the rabbit serum-supplemented bovine albumin polysorbate-80 liquid medium only from the mammary gland of 1 of 4 goats at 13 days after inoculation with serovar hardjo. The positive culture was detected after an 8-month incubation period.


Influence of Solvent in Controlling Peptide-Surface Interactions.
Protein binding to surfaces is an important phenomenon in biology and in modern technological applications. Extensive experimental and theoretical research has been focused in recent years on revealing the factors that govern binding affinity to surfaces. Theoretical studies mainly focus on examining the contribution of the individual amino acids or, alternatively, the binding potential energies of the full peptide, which are unable to capture entropic contributions and neglect the dynamic nature of the system. We present here a methodology that involves the combination of nonequilibrium dynamics simulations with strategic mutation of polar residues to reveal the different factors governing the binding free energy of a peptide to a surface. Using a gold-binding peptide as an example, we show that relative binding free energies are a consequence of the balance between strong interactions of the peptide with the surface and the ability for the bulk solvent to stabilize the peptide.


Effects of dose rate and dose fractionation of irradiation on pulmonary alveolar macrophage colony-forming cells.
We have studied the effects of dose rate and dose fractionation on murine pulmonary alveolar macrophage colony-forming cells (AL-CFC). The dose-response curve of AL-CFC to ionizing irradiation has a Dq of about 100 rad, reflecting the cells' ability to repair sublethal damage. For comparison, we investigated the effect of dose schedule on the committed bone marrow stem cells for both granulocytes and monocytes (GM-CFC) since their dose-response curve has a very small shoulder. We compared the results of dose rates of 3 and 10 rad/min to those obtained with a dose rate of 85 rad/min. We determined survival after giving 100, 300, and 500 rad either in vivo or in vitro. A significant dose rate effect was observed. To study the effect of dose fractionation, a total of 600 rad was given either as a single fraction, three fractions of 200 rad on 3 consecutive days, or six fractions of 100 rad in 3 days. The most dramatic effect was seen in the group that received six 100-rad fractions. No reduction in the number of AL-CFC was seen in this group. In sharp contrast, only a minimal dose schedule effect was observed with GM-CFC.


Synthesis and characterization of pH-sensitive silica nanoparticles for oral-insulin delivery.
The objective of this study is to utilize the pH sensitivity of modified mesoporous silica nanoparticles (MSN) for oral delivery of insulin. Activated silica nanoparticle was modified first with different densities of 3- Chloropropyltrimethoxysilane (3-CTMS) to load the Chlor group which will bond with the organic functional group contained in 4-hydroxybenzoic acid (4-HBA). The nanoparticles were characterized by FTIR and SEM. Equilibrium swelling studies were carried out in enzyme-free simulated gastric and intestinal fluids (SGF and SIF, respectively). Insulin was entrapped in these nanoparticles and the in vitro release profiles were established separately in both (SGF, pH 1) and (SIF, pH 7.4). The amount of loading of insulin was increased with increasing -COO- densities resulting from the increasing number of 4-HBA groups. In these cases, the biological activity of insulin was retained. These results were used to design and improve protein release behavior from these carriers.


Comparative study of dietary phenols with Tartary buckwheat protein (2S/13S): impact on structure, binding sites and functionality of protein.
This research was to investigate the interaction mechanism between 2S albumin and 13S globulin (2S and 13S, the most important storage proteins in Tartary buckwheat seeds) and three phenols (rutin, quercetin and myricetin) regarding the structural and antioxidant properties of their complexes. There are differences in the binding affinity of phenols for 2S and 13S. Rutin had a higher binding affinity for 2S, myricetin had a higher binding affinity for 13S, and 13S exhibited a higher affinity toward phenols than did 2S. Binding with phenols significantly changed the secondary and tertiary structures of 2S and 13S, decreased the surface hydrophobic value and enhanced the antioxidant capacity. Molecular docking and isothermal titration calorimetry showed that the binding processes were spontaneous and that there were hydrogen bonds, hydrophobic bonds and van der Waals force interactions between phenols and proteins. These findings could provide meaningful guidance for the further application of buckwheat protein complex. © 2023 Society of Chemical Industry.


Relation of the tocopherol forms to incident Alzheimer disease and to cognitive change.
High intake of vitamin E from food (tocopherol), but not from supplements (which usually contain alpha-tocopherol), is inversely associated with Alzheimer disease. We examined whether food intakes of vitamin E, alpha-tocopherol equivalents (a measure of the relative biologic activity of tocopherols and tocotrienols), or individual tocopherols would protect against incident Alzheimer disease and cognitive decline over 6 y in participants of the Chicago Health and Aging Project. The 1993-2002 study of community residents aged >or=65 y included the administration of 4 cognitive tests and clinical evaluations for Alzheimer disease. Dietary assessment was by food-frequency questionnaire. Tocopherol intake from food was related to the 4-y incidence of Alzheimer disease determined by logistic regression in 1041 participants who were clinically evaluated (n=162 incident cases) and to change in a global cognitive score determined by mixed models in 3718 participants. Higher intakes of vitamin E (relative risk: 0.74 per 5 mg/d increase; 95% CI: 0.62, 0.88) and alpha-tocopherol equivalents (relative risk: 0.56 per 5 mg/d increase; 95% CI: 0.32, 0.98) were associated with a reduced incidence of Alzheimer disease in separate multiple-adjusted models that included intakes of saturated and trans fats and docosahexaenoic acid. alpha- and gamma-Tocopherol had independent associations. In separate mixed models, a slower rate of cognitive decline was associated with intakes of vitamin E, alpha-tocopherol equivalents, and alpha- and gamma-tocopherols. The results suggest that various tocopherol forms rather than alpha- tocopherol alone may be important in the vitamin E protective association with Alzheimer disease.


Mechanism of inhibition of mammalian DNA topoisomerase I by heparin.
We have previously shown that heparin is a potent inhibitor of a mammalian DNA topoisomerase I. We have now investigated the mechanism of its inhibition. This was carried out first by scrutinizing the structural features of heparin molecules responsible for the inhibition. Commercial heparin preparation was fractionated by antithrombin III-Sepharose into non-adsorbed, low-affinity and high-affinity fractions, of which only the high-affinity fraction of heparin is known to contain a specific oligosaccharide sequence responsible for the binding to antithrombin III. These fractions all exhibited essentially similar inhibitory activities. Furthermore, when chemically sulphated to an extent comparable with or higher than heparin, otherwise inactive glycosaminoglycans such as heparan sulphate, chondroitin 4-sulphate, dermatan sulphate and neutral polysaccharides such as dextran and amylose were converted into potent inhibitors. Sulphated dermatan sulphate, one of the model compounds, was further shown to bind competitively to the same sites on the enzyme as heparin. These observations strongly suggested that topoisomerase inhibition by heparin is attributable primarily, if not entirely, to the highly sulphated polyanionic nature of the molecules. In a second series of experiments we examined whether heparin inhibits only one or both of the topoisomerase reactions, i.e. nicking and re-joining. It was demonstrated that both reactions were inhibited by heparin, but the nicking reaction was more severely affected than was the re-joining reaction.