Physical basis of structural and catalytic Zn-binding sites in proteins.
Zn(2+), an element that is essential to all life forms, can play a catalytic or a solely structural role. Previous works have shown that Zn(2+) binds preferentially to water molecules and His in catalytic sites, but to Cys(-) instructural sites, but the molecular basis for the observed ligand preference is unclear. Here, we show that the different Zn(2+) roles are also reflected in the different bond distances to Zn(2+) in structural and catalytic sites. We reveal the physical basis for the observed differences between structural and catalytic Zn sites: In most catalytic sites, water is found bound to Zn(2+) as it transfers the least charge to Zn(2+) and is less bulky compared to the protein ligands, enabling Zn(2+) to serve as a Lewis acid in catalysis. In most structural sites, however, >/= 2 Cys(-) are found bound to Zn(2+), as Cys(-) transfers the most charge to Zn(2+) and reduces the Zn charge to such an extent that Zn(2+) can no longer act as a Lewis acid; furthermore, steric repulsion among the bulky Cys(S(-)) prevents Zn(2+) from accommodating another ligand. Based on the observed ligand preference and Zn-ligand distance differences between structural and catalytic Zn sites, we present a simple method for distinguishing the two types of sites and for verifying the catalytic role of Zn(2+). Finally, we discuss how the physical bases revealed aid in designing potential drug molecules that target Zn proteins.


The roles of the prostaglandin D(2) receptors DP(1) and CRTH2 in promoting allergic responses.
Prostaglandin D(2) (PGD(2)) is produced by mast cells, Th2 lymphocytes and dendritic cells and has been detected in high concentrations at sites of allergic inflammation. PGD(2) exerts its inflammatory effects through high affinity interactions with the G protein coupled receptors DP(1) and chemoattractant-homologous receptor expressed on Th2 cells (CRTH2, also known as DP(2)). DP(1) and CRTH2 act in concert to promote a number of biological effects associated with the development and maintenance of the allergic response. During the process of allergen sensitization, DP(1) activation may enhance polarization of Th0 cells to Th2 cells by inhibiting production of interleukin 12 by dendritic cells. Upon exposure to allergen in sensitized individuals, activation of DP(1) may contribute to the long lasting blood flow changes in the target organ. CRTH2 is expressed by Th2 lymphocytes, eosinophils and basophils and may mediate the recruitment of these cell types during the late phase allergic response. The role played by CRTH2 in promoting the production of Th2 cytokines and IgE make antagonism of this receptor a particularly attractive approach to the treatment of chronic allergic disease.


In-vitro bio-accessibility and antioxidant activity of commercial standard and enriched whole egg compounds influenced by production and domestic cooking practices.
Commercial whole eggs can be enriched to deliver health-beneficial compounds such as lutein and polyunsaturated fatty acids (PUFA). The combined effect of domestic cooking and gastrointestinal (GI) digestion on enriched egg bio-accessibility and biological activity must be fully elucidated. Thus, this study determines how the combined factors modulate whole egg bio-accessibility and antioxidant activity. Eggs from local supermarkets were cooked and subjected to in vitro GI digestion. The eggs and hydrolysates were characterized for their degree of hydrolysis (DH), carotenoid, PUFA, peptide content, and antioxidant activity. The cooking and digestion influence the DH and carotenoids, whereas PUFA and peptide contents remain unaffected. There was no difference in the antioxidant capacity between the different types of eggs. This study introduces a holistic approach to understanding how production and domestic cooking conditions modulate the bio-accessibility and bioactivity of eggs, which could be used to maximize the health benefits of eggs to consumers.


Improved survival of macroencapsulated islets of Langerhans by preimplantation of the immunoisolating device: a morphometric study.
Encapsulation of cells in a semipermeable membrane may in the future provide an opportunity to treat a variety of endocrine and neurological disorders, without the need for lifelong immunosuppression. The physiological conditions in the device are crucial factors for graft survival. Previously, we have shown that the exchange across the immunoisolating membrane and the microcirculation around the TheraCyte device increase around 3 months after implantation. The aim of this study was to determine whether preimplantation of the TheraCyte device would improve the survival of a later transplanted islet graft. A TheraCyte device was implanted SC on one side of the back of a nondiabetic SD rat. After 3 months, 1500 islets isolated from SD rats were transplanted via the device port. At the same time, another device, loaded with the same number of islets, was implanted on the other side of the back. Both devices were explanted 2 weeks after islet transplantation (i.e., 3.5 months and 0.5 month after device implantation, respectively). Six pairs of devices were evaluated by morphometery. The volume densities of viable islets were 0.22 +/- 0.04 in the preimplanted device vs. 0.06 +/- 0.03 in the nonpreimplanted one (p < 0.05). The corresponding volume densities of fibrosis and necrosis were 0.64 +/- 0.13 vs. 0.85 +/- 0.08 (p < 0.05) and 0.11 +/- 0.14 vs. 0.09 +/- 0.07 (ns), respectively. When the absolute volumes (mm3) were calculated, preimplanted devices contained 1.1 +/- 0.7 endocrine cells while nonpreimplanted ones contained 0.4 +/- 0.2 (p < 0.05). The percentages of insulin- positive beta-cells in the preimplanted versus nonpreimplanted device were 80 +/- 5% and 67 +/- 6%, respectively (p < 0.01). The corresponding volumes of fibrotic tissue were 3.0 +/- 1.8 vs. 5.2 +/- 1.2 (p < 0.05), while the amount of necrotic tissue did not differ significantly (0.42 +/- 0.5 vs. 0.50 +/- 0.3). Preimplantation of the TheraCyte device seems to improve the survival of an encapsulated islet graft and reduce fibroblast outgrowth in the device.


Synthesis, analytical characterization and bioactivity of Ag and Cu nanoparticles embedded in poly-vinyl-methyl-ketone films.
The electrosynthesis of copper and silver core-shell nanoparticles (NPs) by the sacrificial anode technique, employing tetraoctylammonium (TOA) salts as base electrolyte for the first time, is described. These surfactants were selected because they combine high NP stabilizing power with useful disinfecting properties. The resulting colloids were mixed with a solution of an inert dispersing polymer and used to prepare nanostructured composite thin films. The morphologies and chemical compositions of the nanomaterials were characterized by Transmission Electron Microscopy (TEM) and X-ray Photoelectron Spectroscopy (XPS). The TEM reveals that the average core diameter of the metal NPs ranges between 1.7 and 6.3 nm, as a function of the nature of the metal and of the electrosynthesis conditions, and does not change significantly upon inclusion in the polymer matrix. An appreciable concentration of the metal is detected on the nanoparticle surface by XPS. High-resolution XP spectra indicate that both copper and silver are present at zero oxidation state in all of the materials (colloids and composite films). This demonstrates the high efficiency of the surfactant at controlling the morphology and the chemical composition of the nanodispersed metal in both the as-synthesized colloid and in the polymeric dispersion. The nanocoatings are shown to exert a marked inhibitory effect on the growth of eukaryote and prokaryote target microrganisms, and experimental evidence of a synergic disinfecting effect due to the surfactant and the nanodispersed metal is provided. On the basis of these stability and bioactivity results, it is clear that Cu-NPs and Ag-NPs are suitable for application in disinfecting or antifouling paint and coating formulations.


Enzymatic incorporation of an unnatural base pair between 4-propynyl-pyrrole-2-carbaldehyde and 9-methyl-imidazo [(4,5)-b]pyridine into nucleic acids.
A hydrophobic unnatural nucleoside of 4-propynylpyrrole-2-carbaldehyde (designated as Pa') was synthesized to improve its affinity with a pairing partner, 9-methyl-imidazo[(4,5)-b]pyridine (Q), in enzymatic incorporation. In single-nucleotide insertion experiments using the Klenow fragment, the substrate of Pa' (dPa'TP) was efficiently incorporated opposite Q in the template strand, as compared to the incorporation of pyrrole-2-carbaldehyde (dPaTP), which was previously developed.


Long-term follow-up of short intensive multiagent chemotherapy without high-dose methotrexate ('Orange') in children with advanced non-lymphoblastic non-Hodgkin's lymphoma: a children's cancer group report.
Despite prolonged therapy (18 months), children with advanced non-lymphoblastic, non-Hodgkin's lymphoma (NHL) treated on previous Children's Cancer Group (CCG) trials achieved less than a 60% 5-year event-free survival (EFS). In this study we piloted a shorter but more intensive protocol ('Orange') to determine the feasibility, safety, and efficacy of this alternative treatment approach. Thirty-nine children received a CHOP-based induction, etoposide/ifosfamide consolidation, DECAL (dexamethasone, etoposide, cisplatin, cytosine arabinoside (Ara-C) and L-asparaginase) intensification, and either one or two similar but less intense maintenance courses. Patients were stratified to standard-risk (5 months) vs high-risk (7 months) treatment. High risk was defined as either bone marrow disease, CNS disease, mediastinal mass > or = one-third thoracic diameter at T5 and/or LDH > or =2 times institutional upper limits of normal. All other patients were considered to be standard risk. Results were compared with the previous CCG NHL study (CCG-503). Sixteen and 23 patients were considered standard- vs. high-risk, respectively. The 5-year EFS and overall survival (OS) were 77 +/- 7% and 80 +/- 7%, respectively. The 5-year EFS and OS were significantly better in the standard- vs. high-risk subgroups (100% vs. 61 +/- 11%) (P < 0.003) and (100% vs. 65 +/- 11%) (P < 0.01), respectively. Lactate dehydrogenase (LDH) > or =2 x normal (NL) was associated with significantly poorer outcomes (LDH > or =2 x NL vs. <2 x NL) (5-year EFS: 55 +/- 12% vs. 100%) (P < 0.0004). This CCG hybrid regimen, 'Orange', of short and more intensive therapy resulted in a significant improvement in outcomes compared with the previous CCG trial of more prolonged but less intense therapy. This regimen that deletes high-dose methotrexate, if confirmed in a larger trial, could be considered as an alternative treatment approach in children without high tumor burdens (LDH <2 x NL) and Murphy stage III disease.


Inhibitors of the serotonin transporter protein (SERT): the design and synthesis of biotinylated derivatives of 3-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-indoles. High-affinity serotonergic ligands for conjugation with quantum dots.
There is a growing demand for compounds with specificity for the serotonin transporter protein (SERT) that can be conjugated to cadmium selenide/zinc sulfide core shell nanocrystals. This letter describes the design and synthesis of two different biotinylated SERT antagonists that can be attached to streptavidin-coated cadmium selenide/zinc sulfide core shell nanocrystals.


Evaluation of lactic acid bacteria and yeast diversity in traditional white pickled and fresh soft cheeses from the mountain regions of Serbia and lowland regions of Croatia.
The goal of this study was the characterisation of indigenous lactic acid bacteria (LAB) and yeasts isolated from nine white pickled (BG) and nine fresh soft (ZG) artisanal cheeses collected in Serbia and Croatia. While LAB were present in all of the cheeses collected, yeasts were found in all BG cheeses but only in three ZG cheese samples. High LAB and yeast species diversity was determined (average H'(L)=0.4 and H'(Y)=0.8, respectively). The predominant LAB species in white pickled (BG) cheeses were Lactococcus lactis, Lactobacillus plantarum, and Leuconostoc mesenteroides, while in fresh soft (ZG) cheeses the most dominant LAB species were L. lactis, Enterococcus faecalis, and Leuconostoc pseudomesenteroides. Among the 20 yeast species found, Debaryomyces hansenii, Candida zeylanoides, and Torulaspora delbrueckii were found to be predominant in BG cheeses, while Yarrowia lipolytica was predominant in ZG cheeses. The characterisation of metabolic and technological potentials revealed that 53.4% of LAB isolates produced antimicrobial compounds, 44.3% of LAB strains showed proteolytic activity, while most of the yeast species possessed either lipolytic or proteolytic activity. In conclusion, the results obtained in this study showed that the composition of LAB and yeast populations in white pickled and fresh soft cheeses is region specific. The knowledge gained in this study could eventually be used to select region specific LAB and yeast strains for the production of white pickled and fresh soft artisanal cheeses with geographically specific origins under controlled conditions.


Mixing in flowing gas.
We used a mass spectrometer to analyze continuously from a flow stream after changeover of one gas to a mixture of other gases. At the end of a straight cylinder (length = 100 cm, inner diameter = 1/2 inch), light-weight gases appeared later but reached full-scale deflection earlier than heavy gases. Apparently gases with high molecular diffusivity tended to diffuse radially so that they were not carried forward axially in rapidly moving streams. This blunting of the profile of mean concentration vs distance as fresh gas move down a tube, plus spreading of a gradient region of transition between fresh gas and stale gas, are the fundamental processes of diffusion/convection interactions in the lung. Turbulence and molecular diffusion are similar in that they limit the penetration of inspired gases that could occur if laminar flow were the only process acting. However, turbulence and molecular diffusion cause true irreversible mixing of fresh and stale gases, whereas dispersion by laminar flow does not.


Dieulafoy's lesion of esophagus.
Dieulafoy's lesion is a rare arterial malformation that can cause massive gastrointestinal hemorrhage. The lesion occurs most commonly in the proximal stomach. The esophagus is not a common location for this lesion. We present the case of a 25-year-old woman who was admitted to our emergency unit with the findings of hematemesis and melena. Early upper gastrointestinal endoscopic examination revealed a Dieulafoy's lesion, which was located in the distal esophagus. Endoscopic band ligation stopped the bleeding successfully. The patient was discharged 3 days after the band ligation without any complications. Dieulafoy's lesion may cause severe, life-threatening bleeding. Endoscopic diagnosis can be difficult because of the small size and obscure location of the lesion. An abnormally dilated artery that penetrates through the mucosa constitutes the etiology. Endoscopy plays an important role in the diagnosis and treatment of this pathology. Despite widespread awareness of this entity, it may present a real challenge for the endoscopist due to the small size and hidden location of the lesion. The endoscopic approach to occult gastrointestinal bleeding for the diagnosis of vascular malformations is accepted as a quick and safe diagnostic method.


The migration of lymphocytes into rat popliteal lymph nodes during antigenic promotion or competition between heterologous erythrocytes.
The injection of chicken and sheep red blood cells (CRBC and SRBC) into rat popliteal lymph nodes either together or sequentially 2, 4, 6, or 8 days apart resulted in an enhanced immune response when the second antigen was injected 2 or 4 days after the injection of the first antigen (antigenic promotion) or a suppressed immune response when the second antigen was injected 6 days after the injection of the first antigen (antigenic competition). The immune response to either antigen was dependent upon the time of administration of the second antigen with respect to the first antigen. Lymphocyte migration into antigenically stimulated lymph nodes was greater when the two antigens were injected sequentially rather than together. Further, the migration of lymphocytes into the lymph node was enhanced when the second antigen was injected during the inductive or suppressive phase of the immune response to the first antigen (CRBC) regardless of whether the same (CRBC) or an antigenically unrelated antigen (SRBC) was used as the second antigen. While antigenic promotion may in part be explained by the increased rate at which lymphocytes migrate into lymph nodes, lymphocyte migration is also enhanced during antigenic competition. This suggests that while suppressor cells/factors may regulate the effector phase of an immune response they do not directly modulate the migration of blood-borne lymphocytes into the lymph node.


Inconsistency in the Self-report of Chronic Diseases in Panel Surveys: Developing an Adjudication Method for the Health and Retirement Study.
Chronic disease data from longitudinal health interview surveys are frequently used in epidemiologic studies. These data may be limited by inconsistencies in self-report by respondents across waves. We examined disease inconsistencies in the Health and Retirement Study and investigated a multistep method of adjudication. We hypothesized a greater likelihood of inconsistences among respondents with cognitive impairment, of underrepresented race/ethnic groups, having lower education, or having less income/wealth. We analyzed Waves 1995-2010, including adults 51 years and older (N = 24,156). Diseases included hypertension, heart disease, lung disease, diabetes, cancer, stroke, and arthritis. We used questions about the diseases to formulate a multistep adjudication method to resolve inconsistencies across waves. Thirty percent had inconsistency in their self-report of diseases across waves, with cognitive impairment, proxy status, age, Hispanic ethnicity, and wealth as key predictors. Arthritis and hypertension had the most frequent inconsistencies; stroke and cancer, the fewest. Using a stepwise method, we adjudicated 60%-75% of inconsistent responses. Discrepancies in the self-report of diseases across multiple waves of health interview surveys are common. Differences in prevalence between original and adjudicated data may be substantial for some diseases and for some groups, (e.g., the cognitively impaired).


Movement patterns of a clear-wing hawkmoth, Hemaris fuciformis, foraging at red catchfly, Viscaria vulgaris.
The bee hawkmoth, Hemaris fuciformis, tended to fly in the same direction between successive visits to inflorescences of Viscaria vulgaris. Upon leaving an inflorescence it did not fly to the first inflorescence in its path, but to the second. At inflorescences, the number of animals that started probing decreased from the bottom to the top. These movement patterns probably serve to reduce the risk of revisiting flowers. The movements of the moths at inflorescences with 2 flowers per tier were investigated and related to the nectar distribution. The nectar is distributed according to a "bonanza-blank" pattern, flowers being either empty or full, probably because previous visitors have emptied some, but not all, flowers. It is shown that the two flowers of a tier more often than random belong to the category: blank-bonanza. Thus, if a blank is found, the animal should move sideways to the second flower at the same tier in order to find a bonanza. The flowers of the inflorescences tend to be arranged in vertical rows, and it is shown that two flowers, one above the other, more often than random belong to the two categories: blank-blank or bonanza-bonanza. Thus, if a bonanza is found, the animal should move up to the flower above, in order to find another bonanza. Rules of movement at inflorescences and a rule of departure are proposed based on the probability distribution of the nectar. The observed movements agree quite well with the predicted ones.


Toxicity assessment of Clostridium difficile toxins in rodent models and protection of vaccination.
Clostridium difficile is the leading cause of hospital-acquired diarrhea, also known as C. difficile associated diarrhea. The two major toxins, toxin A and toxin B are produced by most C. difficile bacteria, but some strains, such as BI/NAP1/027 isolates, produce a third toxin called binary toxin. The precise biological role of binary toxin is not clear but it has been shown to be a cytotoxin for Vero cells. We evaluated the toxicity of these toxins in mice and hamsters and found that binary toxin causes death in both animals similar to toxins A and B. Furthermore, immunization of mice with mutant toxoids of all three toxins provided protection upon challenge with native toxins. These results support the concept that binary toxin contributes to the pathogenicity of C. difficile and provide a method for monitoring the toxicity of binary toxin components in vaccines.


The Timing of Pregnancies After Bariatric Surgery has No Impact on Children's Health-a Nationwide Population-based Registry Analysis.
Bariatric surgery has a favorable effect on fertility in women. However, due to a lack of data regarding children's outcomes, the ideal time for conception following bariatric surgery is unknown. Current guidelines advise avoiding pregnancy during the initial weight loss phase (12-24 months after surgery) as there may be potential risks to offspring. Thus, we aimed to analyze health outcomes in children born to mothers who had undergone bariatric surgery. The surgery-to-delivery interval was studied. A nationwide registry belonging to the Austrian health insurance funds and containing health-related data claims was searched. Data for all women who had bariatric surgery in Austria between 01/2010 and 12/2018 were analyzed. A total of 1057 women gave birth to 1369 children. The offspring's data were analyzed for medical health claims based on International Classification of Diseases (ICD) codes and number of days hospitalized. Three different surgery-to-delivery intervals were assessed: 12, 18, and 24 months. Overall, 421 deliveries (31%) were observed in the first 2 years after surgery. Of these, 70 births (5%) occurred within 12 months after surgery. The median time from surgery to delivery was 34 months. Overall, there were no differences noted in frequency of hospitalization and diagnoses leading to hospitalization in the first year of life, regardless of the surgery-to-delivery interval. Pregnancies in the first 24 months after bariatric surgery were common. Importantly, the surgery-to-delivery interval had no significant impact on the health outcome of the children.


Most Trial Eligibility Criteria and Patient Baseline Characteristics Do Not Modify Treatment Effect in Trials Using Targeted Therapies for Rheumatoid Arthritis: A Meta-Epidemiological Study.
To determine if variations in trial eligibility criteria and patient baseline characteristics could be considered effect modifiers of the treatment response when testing targeted therapies (biological agents and targeted synthetic disease modifying antirheumatic drugs (DMARDs)) for rheumatoid arthritis (RA). We conducted a meta-epidemiological study of all trials evaluating a targeted therapy approved by regulatory authorities for treating RA. The database search was completed on December 11th 2013. Eligible trials reported ACR20 data at months 3-6 and used an add-on design. Odds ratios (ORs) were calculated from the response rates and compared among the trial eligibility criteria/patient baseline characteristics of interest. Comparisons are presented as the Ratio of Odds Ratios (ROR). Sixty-two trials (19,923 RA patients) were included in the primary analyses using ACR20 response. Overall, targeted therapies constituted an effective treatment (OR 3.96 95% confidence interval (CI) 3.41 to 4.60). The majority of the trial eligibility criteria and patient baseline characteristics did not modify treatment effect. The added benefit of targeted therapies was lower in trials including "DMARD-naïve" patients compared with trials including "DMARD inadequate responders" (ROR = 0.45, 95%CI 0.31 to 0.66) and trials including "targeted therapy inadequate responders" (0.50, 95%CI 0.29 to 0.87), test for interaction: p = 0.0002. Longer mean disease duration was associated with a higher likelihood of responding to treatment (β = 1.05, 95%CI 1.00 to 1.11 OR's per year; p = 0.03). Analyses conducted using DAS28-remission as the outcome supported the above-mentioned findings. Our results suggest that a highly selective inclusion is not associated with greater treatment effect, as might otherwise be expected. The added benefit of a targeted therapy was lower in trials including patients who were DMARD-naïve and trials including patients with shorter disease durations.


[Behavior of selected oxidoreductases in histochemical and electro- phoretic studies of the rat kidney after administration of Biseptol 480 (co-trimoxazole)].
Examinations of the rat kidney metabolism were carried out by histological (staining with haematoxylin and eosin), histochemical (response to lactate and succinic dehydrogenases activities), and electrophoretic methods (separation of isoenzymes of lactate dehydrogenase) after treating animals with the sulphonamide Biseptol 480, for 7 (consecutive) d by stomach tube. Biseptol given to the animals exceeded 10 times human therapeutical dose. It was found out that Biseptol 480, applied to the animals with healthy urinary system, does not pathologically disturb the examined stages of the metabolic processes in the rat's kidney.


Impaired glucose tolerance in the U.S. population.
Impaired glucose tolerance (IGT) constitutes two-thirds of all glucose intolerance in the United States and is a major risk factor for diabetes. Despite these findings, the clinical and epidemiological significance of IGT has not been well investigated. The Second National Health and Nutrition Examination Survey, a cross-sectional study in which 75-g 2-h oral glucose tolerance tests (OGTTs) were performed, has provided an opportunity to examine the characteristics of IGT in the U.S. population. Data from the survey have been extrapolated to represent all U.S. residents. The findings indicate that approximately 11.2% of Americans aged 20-74 yr have IGT compared to 6.6% with diabetes. Rates of IGT increased with age for White men and women and Black men but declined for Black women greater than 54 yr of age, possibly because greater obesity in Black women precipitated earlier conversion of IGT to diabetes. The distribution of 2-h glucose values showed IGT to be part of a continuum of glucose intolerance extending from normal to diabetes. Individuals with IGT had rates of risk factors for non-insulin-dependent diabetes (age, plasma glucose, past obesity, family history of diabetes, physical inactivity) that were intermediate between those of individuals with normal glucose tolerance and those with diabetes, although current obesity was similar for IGT and diabetes. The proportion of people with medical histories of diabetes-related conditions did not differ between IGT and normal glucose tolerance. However, several cardiovascular findings were more prevalent in individuals with IGT than in those with normal glucose tolerance, including hypertension, serum cholesterol, angina, abnormal heart findings, and medical history of arteriosclerosis and stroke. Both obesity and reported family history of diabetes were associated with higher rates of IGT, with the effect of weight gain on the prevalence of IGT occurring at lower levels than for diabetes.


Thoracoscopy: a useful tool in the diagnosis of thoracic disease.
During a 12-month period, 11 patients underwent diagnostic thoracoscopy for previously undiagnosed thoracic disease. In all patients, the diagnosis had been unobtainable by the usual diagnostic modalities of bronchoscopy, scalene node biopsy, mediastinoscopy, thoracentesis, or closed pleural biopsy. Thoracoscopy was diagnostic in 10 of the 11 patients. There was no morbidity or mortality. In all patients the indication for thoracoscopy was suspected malignancy. The majority of patients had recurrent pleural effusions in which routine cytological studies and tissue biopsies had been nondiagnostic. Pathological findings were mesothelioma in 3 patients, primary carcinoma of the lung in 4, congestive heart failure with pleural effusion in 1, metastatic carcinoma in 2, and inflammatory disease in 1 patient. Indications, techniques, and results are discussed. Thoracoscopy is a valuable tool in the diagnosis of thoracic disease; with it, unneccessary thoracotomy can often be avoided.


Chromosome tests with 134 compounds on Chinese hamster cells in vitro--a screening for chemical carcinogens.
Chromosomal aberration tests in vitro were carried out on Chinese hamster cells grown in culture with various chemicals, including carcinogenic N-nitroso compounds and their related derivatives, food additives, medical drugs, pesticides and other chemicals commonly used in laboratories or industries. Sixty-three of the 134 chemicals gave negative results in our test system even with doses at which the cell growth was markedly inhibited. Nearly all compounds known to be mutagenic in bacteria were also positive in our tests. Both urethane and diethylstilbestrol were positive, even though they are known to be carcinogenic but not mutagenic in bacteria. Compounds such as N-alkyl-N'-nitroguanidines, barbital, sodium benzoate, saccharin sodium, sodium nitrite, sodium nitrate and 4-aminoquinoline-1-oxide were positive in our chromosome tests, but they have not been conclusively tested for their carcinogenicity.


Assessment and modeling of the novel toxicity data set of phenols to Chlorella vulgaris.
This study provides for the first time the 96 h toxicity of 30 phenols to Chlorella vulgaris. Analysis of the novel data set revealed that the algal toxicity of polar narcotics and respiratory uncouplers was correlated strongly to the pH corrected hydrophobicity parameter, Log D, demonstrating the importance of ionization in the C. vulgaris test system. Compounds expected to act by more reactive mechanisms were shown to have toxicity in excess of that predicted by Log D and were successfully modelled using the activation energy index (AEI). Three global quantitative structure-activity relationships (QSARs) were constructed using the C. vulgaris data set and validated externally using a data set retrieved from literature comprising the toxicity of 58 compounds to freshwater alga Pseudokirchneriella subcapitata. Results revealed that the response-surface model was highly interpretable and provided acceptable predictions for polar narcotics and respiratory uncouplers, though it lacked the reliability to predict the toxicity of reactive phenols. In two other externally validated QSAR models, a WHIM (Weighted Holistic Invariant Molecular) descriptor, namely, Tm (T total size index/weighted by atomic masses), revealed promising results that could be used to predict algal toxicity of compounds other than phenols such as anilines.


Lipid raft disarrangement as a result of neuropathological progresses: a novel strategy for early diagnosis?
Lipid rafts are the preferential site of numerous membrane signaling proteins which are involved in neuronal functioning and survival. These proteins are organized in multiprotein complexes, or signalosomes, in close contact with lipid classes particularly represented in lipid rafts (i.e. cholesterol, sphingolipids and saturated fatty acids), which may contribute to physiological responses leading to neuroprotection. Increasing evidence indicates that alteration of lipid composition in raft structures as a consequence of neuropathologies, such as Alzheimer's disease (AD) and Parkinson's disease (PD), causes a dramatic increase in lipid raft order. These phenomena may correlate with perturbation of signalosome activities, likely contributing to neurodegenerative progression. Interestingly, significant disruption of stable raft microenvironments has been already observed in the first stages of either AD or PD, suggesting that these alterations may represent early events in the neuropathological development. In this regard, the search for biochemical markers, such as specific metabolic products altered in the brain at the first steps of the disease, presently represents an important challenge for early diagnostic strategies. Alterations of these biomarkers may be reflected in either plasma or cerebrospinal fluid, thus representing a potential strategy to predict an accurate diagnosis. We propose that pathologically-linked lipid raft markers may be interesting candidates to be explored at this level, although it has not been studied so far to what extent alteration of different signalosome components may be reflected in peripheral fluids. In this mini-review, we will discuss on relevant aspects of lipid rafts that contribute to the modulation of neuropathological events related to AD and PD. An interesting hypothesis is that anomalies on raft biomarkers measured at peripheral fluids might mirror the lipid raft pathology observed in early stages of AD and PD.


Retinal AAV8-RS1 Gene Therapy for X-Linked Retinoschisis: Initial Findings from a Phase I/IIa Trial by Intravitreal Delivery.
This study evaluated the safety and tolerability of ocular RS1 adeno-associated virus (AAV8-RS1) gene augmentation therapy to the retina of participants with X-linked retinoschisis (XLRS). XLRS is a monogenic trait affecting only males, caused by mutations in the RS1 gene. Retinoschisin protein is secreted principally in the outer retina, and its absence results in retinal cavities, synaptic dysfunction, reduced visual acuity, and susceptibility to retinal detachment. This phase I/IIa single-center, prospective, open-label, three-dose-escalation clinical trial administered vector to nine participants with pathogenic RS1 mutations. The eye of each participant with worse acuity (≤63 letters; Snellen 20/63) received the AAV8-RS1 gene vector by intravitreal injection. Three participants were assigned to each of three dosage groups: 1e9 vector genomes (vg)/eye, 1e10 vg/eye, and 1e11 vg/eye. The investigational product was generally well tolerated in all but one individual. Ocular events included dose-related inflammation that resolved with topical and oral corticosteroids. Systemic antibodies against AAV8 increased in a dose-related fashion, but no antibodies against RS1 were observed. Retinal cavities closed transiently in one participant. Additional doses and immunosuppressive regimens are being explored to pursue evidence of safety and efficacy (ClinicalTrials.gov: NCT02317887).


Comparison of various anthropometric and body fat indices in identifying cardiometabolic disturbances in Chinese men and women.
Although many adiposity indices may be used to predict obesity-related health risks, uncertainty remains over which of them performs best. This study compared the predictive capability of direct and indirect adiposity measures in identifying people at higher risk of metabolic abnormalities. This population-based cross-sectional study recruited 2780 women and 1160 men. Body weight and height, waist circumference (WC), and hip circumference (HC) were measured and body mass index (BMI), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR) were calculated. Body fat (and percentage of fat) over the whole body and the trunk were determined by bioelectrical impedance analysis (BIA). Blood pressure, fasting lipid profiles, and glucose and urine acid levels were assessed. In women, the ROC and the multivariate logistic regression analyses both showed that WHtR consistently had the best performance in identifying hypertension, dyslipidemia, hyperuricemia, diabetes/IFG, and metabolic syndrome (MetS). In men, the ROC analysis showed that WHtR was the best predictor of hypertension, WHtR and WC were equally good predictors of dyslipidemia and MetS, and WHtR was the second-best predictor of hyperuricemia and diabetes/IFG. The multivariate logistic regression also found WHtR to be superior in discriminating between MetS, diabetes/IFG, and dyslipidemia while BMI performed better in predicting hypertension and hyperuricemia in men. The BIA-derived indices were the second-worst predictors for all of the endpoints, and HC was the worst. WHtR was the best predictor of various metabolic abnormalities. BMI may be used as an alternative measure of obesity for identifying hypertension in both sexes.


Low-dose contrast-enhanced time-resolved MR angiography at 3T: diagnostic accuracy for treatment planning and follow-up of vascular malformations.
To assess the accuracy of low-dose contrast-enhanced time-resolved 3T magnetic resonance angiography (MRA) for the morphological and functional assessment of vascular malformations (VM), and to evaluate its diagnostic potential for the depiction of treatment-induced changes. Twenty-five patients with known VM underwent MRA to evaluate the location and extent of lesions and their haemodynamic characteristics. Three-dimensional (3D) T1-weighted time-resolved sequences were acquired following the administration of 0.05mmol/kg of gadobenate dimeglumine. VM were classified according to their morphology and haemodynamic characteristics. All patients thereafter underwent conventional angiography to confirm the diagnosis and to treat the lesions (embolization or sclerotherapy). Follow-up MRA was performed 30 days after treatment to assess morphological and functional changes. A visual analogue scale (VAS) was used to clinically assess the severity of symptoms before and after therapy. Based on haemodynamic characteristics, VM were classified as predominantly arterial [4 (16%)], artero-venous [19 (76%)] or venous [2 (8%)]. Twenty-three (92%) lesions were classified as high-flow VM and two (8%) as low-flow VM. Intralesional thrombosis was present in 17 (68%) lesions before therapy and in 10 lesions (40%) after therapy. The median VAS scores were 5±1 before treatment and 4±2 after treatment. Very good correlation (Spearman's correlation coefficient: rho=0.87; p=0.000) was noted between the reduction of lesion size on follow-up MRA and pain relief as assessed by VAS. Low-dose contrast-enhanced time-resolved 3T MRA can be used to define morphological and functional aspects of VM accurately during treatment planning and follow-up, and can identify post-therapy changes that positively correlate with treatment outcome.


What accounts for the relationship between social class and smoking cessation? Results of a path analysis.
Despite the overall decline in cigarette smoking prevalence in the US, social class inequalities in smoking are likely to persist, or even to widen. One possible reason for the increasing gap in smoking prevalence across social class could be our lack of understanding of causal mechanisms: in other words, what accounts for the social gradient in smoking behavior? In this paper, we examine the mechanisms behind social gradients related to smoking cessation by use of path analysis techniques. The data come from a 3-year follow-up telephone survey of a cohort of US adults. The sample for the present analysis was drawn from the 481 respondents who reported being smokers and employed at baseline and who completed the follow-up interview. We examined two social class indicators, educational attainment and household income, in relation to smoking cessation. We tested the potential mediating effects of the following variables: differential use of resources for smoking cessation (e.g., booklet, pamphlet, quit line, nicotine replacement therapy and smoking cessation program), differential environments in terms of smoking at worksite and home, and differences in peer smoking. Our path analyses suggest that smokers from high social class are likely to use effective resources for smoking cessation and have restrictive home environment in terms of smoking, which leads to a relatively higher smoking cessation rate compared to those from low social class. The results of this study suggest that interventions should target resources for smoking cessation and home environments in terms of smoking to reduce socio-economic disparities in smoking cessation.


Effectiveness and safety of pemetrexed-based doublet versus pemetrexed alone as second-line treatment for advanced non-small-cell lung cancer: a systematic review and meta-analysis.
To compared pemetrexed-based doublet with single-agent pemetrexed as second-line treatment for advanced non-small-cell lung cancer We systematically searched for randomized clinical trials that compared pemetrexed-based doublet with single-agent pemetrexed in patients with histologically proven non-small-cell lung cancer. The primary end point was overall survival. Secondary end points were progression-free survival, overall response rate and grade 3 or 4 toxicity. Data were extracted from the studies by 2 independent reviewers. The meta-analysis was performed by Stata version 10.0 software (Stata Corporation, College Station, Texas, USA). Five randomized clinical trials (totally 1,186 patients) were eligible. Meta-analysis showed that there was significant improvement in PFS (HR 0.82, 95% CI 0.71-0.95, P = 0.007) and overall response rate (OR 2.39, 95% CI 1.58-3.62, P = 0.000) in pemetrexed-based doublet group, compared with pemetrexed alone, though the pooled HR for overall survival (HR 0.89, 95% CI 0.76-1.04; P = 0.129) showed no significant difference between the two groups. However, there were more incidences of grade 3 or 4 neutropenia (OR 2.3, 95% CI 1.4-3.77, P = 0.001), thrombocytopenia (OR 6.41, 95% CI 2.57-16.0, P = 0.000), and leucopenia (OR 2.45, 95% CI 1.13-5.34, P = 0.024) in pemetrexed-based doublet group. With regard to the risk of grade 3 or 4 anemia (OR 0.71, 95% CI 0.17-2.91, P = 0.629) and fatigue (OR 1.47, 95% CI 0.92-2.35, P = 0.104), there was no significant difference between the two groups. Pemetrexed-based doublet therapy didn't gain any benefit in survival but significantly improved PFS and better ORR compared with single-agent pemetrexed as second-line therapy for advanced non-small-cell lung cancer. However, more incidences of grade 3 or 4 neutropenia, thrombocytopenia, and leucopenia were observed in pemetrexed-based doublet group.


The RPA-RNF20-SNF2H cascade promotes proper chromosome segregation and homologous recombination repair.
The human tumor suppressor Ring finger protein 20 (RNF20)-mediated histone H2B monoubiquitination (H2Bub) is essential for proper chromosome segregation and DNA repair. However, what is the precise function and mechanism of RNF20-H2Bub in chromosome segregation and how this pathway is activated to preserve genome stability remain unknown. Here, we show that the single-strand DNA-binding factor Replication protein A (RPA) interacts with RNF20 mainly in the S and G2/M phases and recruits RNF20 to mitotic centromeres in a centromeric R-loop-dependent manner. In parallel, RPA recruits RNF20 to chromosomal breaks upon DNA damage. Disruption of the RPA-RNF20 interaction or depletion of RNF20 increases mitotic lagging chromosomes and chromosome bridges and impairs BRCA1 and RAD51 loading and homologous recombination repair, leading to elevated chromosome breaks, genome instability, and sensitivities to DNA-damaging agents. Mechanistically, the RPA-RNF20 pathway promotes local H2Bub, H3K4 dimethylation, and subsequent SNF2H recruitment, ensuring proper Aurora B kinase activation at centromeres and efficient loading of repair proteins at DNA breaks. Thus, the RPA-RNF20-SNF2H cascade plays a broad role in preserving genome stability by coupling H2Bub to chromosome segregation and DNA repair.


Cancer deaths and cases attributable to lifestyle factors and infections in China, 2013.
The burden of cancer in China is high, and it is expected to further increase. Information on cancers attributable to potentially modifiable risk factors is essential in planning preventive measures against cancer. We estimated the number and proportion of cancer deaths and cases attributable to ever-smoking, second-hand smoking, alcohol drinking, low fruit/vegetable intake, excess body weight, physical inactivity, and infections in China, using contemporary data from nationally representative surveys and cancer registries. The number of cancer deaths and cases in 2013 were obtained from the National Central Cancer Registry of China and data on most exposures were obtained from the China National Nutrition and Health Survey 2002 or 2006 and Global Adult Tobacco Smoking 2010. We used a bootstrap simulation method to calculate the number and proportion of cancer deaths and cases attributable to risk factors and their corresponding 95% confidence intervals (CIs), allowing for uncertainty in data. Approximately 718 000 (95% CI 702 100-732 200) cancer deaths in men and 283 100 (278 800-288 800) cancer deaths in women were attributable to the studied risk factors, accounting for 52% of all cancer deaths in men and 35% in women. The numbers for incident cancer cases were 952 500 (95% CI 934 200-971 400) in men and 442 700 (437 200-447 900) in women, accounting for 47% of all incident cases in men and 28% in women. The greatest proportions of cancer deaths attributable to risk factors were for smoking (26%), HBV infection (12%), and low fruit/vegetable intake (7%) in men and HBV infection (7%), low fruit/vegetable intake (6%), and second-hand smoking (5%) in women. Effective public health interventions to eliminate or reduce exposure from these risk factors, notably tobacco control and vaccinations against carcinogenic infections, can have considerable impact on reducing the cancer burden in China.


Change in phase relations of circadian rhythms in cell proliferation induced by time-limited feeding in BALB/c X DBA/2F1 mice bearing a transplantable Harding-Passey tumor.
The synchronization of the circadian rhythms of [3H]thymidine uptake (as gauge of DNA synthesis and presumably of cell proliferation) in colon, thymus, and Harding-Passey melanoma were studied in 456 male BALB/c X DBA/2F1 mice under a 12-hr-light, 12-hr-dark regimen. In two groups of animals, the feeding time was restricted to 4 hr/day (either at the beginning of the light span or at the beginning of the dark span). The circadian rhythms in body temperature and [3H]thymidine uptake in the colon were determined in their timing primarily by the time of food intake. In contrast, the circadian rhythm of [3H]thymidine uptake in the thymus and in the transplanted melanoma remained synchronized with the lighting regimen, and under the conditions of this study, was not altered in its timing by the change in feeding time. It thus appears feasible to alter the phase relations between certain circadian rhythms of host and tumor. If applicable to the human situation, this observation might be of interest for the scheduling of chemo- and radiotherapy, in an attempt to obtain maximal effects upon the tumor with minimal undesired side effects upon vital functions of the host.


Endoscopic transmural debridement of symptomatic organized pancreatic necrosis (with videos).
Surgical management of pancreatic necrosis is associated with significant morbidity and mortality. Several weeks after an episode of a necrotizing pancreatitis, necrosis can become organized. By the time necrosis becomes organized, endoscopic therapy has the potential to offer an alternative treatment to surgery. To evaluate the safety and efficacy of endoscopic debridement of organized pancreatic necrosis and to identify procedural aspects that may improve outcome. Retrospective cohort study. Tertiary referral center. All consecutive patients who underwent this novel endoscopic approach were included. Treatment started with a cystoenterostomy or a cystogastrostomy. The next steps consisted of balloon dilation, up to 18 mm; advancement of an endoscope into the retroperitoneal cavity; and endoscopic debridement of the collection under direct endoscopic vision. Debridement was repeated every 2 days until most necrotic material was evacuated. In addition, nasocystic catheter irrigation was performed manually with saline solution 6 to 8 times a day. Clinical success, number of endoscopic procedures, and complications. Twenty-five patients were identified, who had undergone debridement of 27 collections. In 11, 13, 2, and 1 collections, 1, 2, 3, and 4 endoscopic debridement procedures, respectively, were performed. There was no mortality. Severe complications that required surgery occurred in 2 patients: hemorrhage in 1 case and perforation of cyst wall in the other. During a median follow-up of 16 months (range 3-38 months), the overall clinical success rate with resolution of the collection and related symptoms was 93%. Retrospective study. In this study, we showed that endoscopic debridement is an effective and relatively safe minimally invasive therapy in patients with symptomatic organized pancreatic necrosis. Further comparative studies are warranted to define its definitive role in the management of these patients.


Which elderly newly diagnosed glioblastoma patients can benefit from radiotherapy and temozolomide? A PERNO prospective study.
The role of temozolomide concurrent with and adjuvant to radiotherapy (RT/TMZ) in elderly patients with glioblastoma (GBM) remains unclear. We evaluated the outcome of patients >70 years in the context of the Project of Emilia-Romagna Region in Neuro-Oncology (PERNO), the first Italian prospective observational population-based study in neuro-oncology. For this analysis the criteria for selecting patients enrolled in the PERNO study were: age >70 years; PS 0-3; histologically confirmed GBM; postoperative radiotherapy (RT) after surgery with or without concomitant temozolomide (TMZ) or postsurgical TMZ alone. Between January 2009 and December 2010, 76 GBM elderly patients were identified in the prospective PERNO study. Twenty-three patients did not receive any treatment after surgery, and 53 patients received postsurgical treatments (25 patients received RT alone and 28 patients RT/TMZ). Median survival was 11.1 months (95 % CI 8.8-13.5), adding temozolomide concomitant and adjuvant to radiotherapy it was 11.6 months (95 % CI 8.6-14.6), and 9.3 months (95 % CI 8.1-10.6) in patients treated with RT alone (P = 0.164). However, patients with MGMT methylated treated with RT/TMZ obtained a better survival (17.2 months, 95 % CI 11.5-22.9) (P = 0.042). No difference in terms of survival were observed if patients with MGMT unmethylated tumor received RT alone, or RT/TMZ or, in MGMT methylated tumor, if patients received radiotherapy alone. In elderly patients RT/TMZ represent a widely used approach but it is effective with methylated MGMT tumors only.


Conserved function of the lysine-based KXD/E motif in Golgi retention for endomembrane proteins among different organisms.
We recently identified a new COPI-interacting KXD/E motif in the C-terminal cytosolic tail (CT) of Arabidopsis endomembrane protein 12 (AtEMP12) as being a crucial Golgi retention mechanism for AtEMP12. This KXD/E motif is conserved in CTs of all EMPs found in plants, yeast, and humans and is also present in hundreds of other membrane proteins. Here, by cloning selective EMP isoforms from plants, yeast, and mammals, we study the localizations of EMPs in different expression systems, since there are contradictory reports on the localizations of EMPs. We show that the N-terminal and C-terminal GFP-tagged EMP fusions are localized to Golgi and post-Golgi compartments, respectively, in plant, yeast, and mammalian cells. In vitro pull-down assay further proves the interaction of the KXD/E motif with COPI coatomer in yeast. COPI loss of function in yeast and plants causes mislocalization of EMPs or KXD/E motif-containing proteins to vacuole. Ultrastructural studies further show that RNA interference (RNAi) knockdown of coatomer expression in transgenic Arabidopsis plants causes severe morphological changes in the Golgi. Taken together, our results demonstrate that N-terminal GFP fusions reflect the real localization of EMPs, and KXD/E is a conserved motif in COPI interaction and Golgi retention in eukaryotes.


An entrant of smaller fullerene: C56 captured by chlorines and aligned in linear chains.
A smaller fullerene C56 (#913) is stabilized, isolated, and crystallographically characterized as C56Cl10. The geometric parameters of C56Cl10 imply the otherwise unstable cage of C56 can be stabilized by chlorination through releasing its surface strains and maintaining fragmental aromaticity. An unexpected C Cl...ClC short contact, as well as the linear alignment with pearl-necklace-shaped, is revealed in C56Cl10 crystal.


Betulinic acid-induced mitochondria-dependent cell death is counterbalanced by an autophagic salvage response.
Betulinic acid (BetA) is a plant-derived pentacyclic triterpenoid that exerts potent anti-cancer effects in vitro and in vivo. It was shown to induce apoptosis via a direct effect on mitochondria. This is largely independent of proapoptotic BAK and BAX, but can be inhibited by cyclosporin A (CsA), an inhibitor of the permeability transition (PT) pore. Here we show that blocking apoptosis with general caspase inhibitors did not prevent cell death, indicating that alternative, caspase-independent cell death pathways were activated. BetA did not induce necroptosis, but we observed a strong induction of autophagy in several cancer cell lines. Autophagy was functional as shown by enhanced flux and degradation of long-lived proteins. BetA-induced autophagy could be blocked, just like apoptosis, with CsA, suggesting that autophagy is activated as a response to the mitochondrial damage inflicted by BetA. As both a survival and cell death role have been attributed to autophagy, autophagy-deficient tumor cells and mouse embryo fibroblasts were analyzed to determine the role of autophagy in BetA-induced cell death. This clearly established BetA-induced autophagy as a survival mechanism and indicates that BetA utilizes an as yet-undefined mechanism to kill cancer cells.


Effects of indoleamine 2,3-dioxygenases in carbon tetrachloride-induced hepatitis model of rats.
Indoleamine 2,3-dioxygenase (IDO) converts tryptophan to l-kynurenine, and it is noted as a relevant molecule in promoting tolerance and suppressing adaptive immunity. In this study, to investigate the effects of IDO in carbon tetrachloride (CCl(4) )-induced hepatitis model, the levels of IDO enzymic activities in the mock group, the control group and the 1-methyl-D-tryptophan (1-MT)-treated group were confirmed by determination of l-kynurenine concentrations. Serum alanine aminotransferase levels in 1-MT-treated rats after CCl(4) injection significantly increased compared with those in mock and control groups. In CCl(4)-induced hepatitis models, tumour necrosis factor-α (TNF-α) is critical in the development of liver injury. The mRNA expression and secretion levels of TNF-α in the liver from 1-MT-treated rats were more enhanced compared with those in the mock and the control groups. Moreover, the levels of cytokine and chemokine from mock, control group and 1-MT-treated rats after treated with CCl(4) were analyzed by ELISA, and the level of interleukin-6 was found to increase in 1-MT-treated rats. It was concluded that the deficiency of IDO exacerbated liver injury in CCl(4)-induced hepatitis and its effect may be connected with TNF-α and interleukin-6.


A standardized extract of Butea monosperma (Lam.) flowers suppresses the IL-1β-induced expression of IL-6 and matrix-metalloproteases by activating autophagy in human osteoarthritis chondrocytes.
Osteoarthritis (OA) is a leading cause of joint dysfunction, disability and poor quality of life in the affected population. The underlying mechanism of joint dysfunction involves increased oxidative stress, inflammation, high levels of cartilage extracellular matrix degrading proteases and decline in autophagy-a mechanism of cellular defense. There is no disease modifying therapies currently available for OA. Different parts of the Butea monosperma (Lam.) plant have widely been used in the traditional Indian Ayurvedic medicine system for the treatment of various human diseases including inflammatory conditions. Here we studied the chondroprotective effect of hydromethanolic extract of Butea monosperma (Lam.) flowers (BME) standardized to the concentration of Butein on human OA chondrocytes stimulated with IL-1β. The hydromethanolic extract of Butea monosperma (Lam.) (BME) was prepared with 70% methanol-water mixer using Soxhlet. Chondrocytes viability after BME treatment was measured by MTT assay. Gene expression levels were determined by quantitative polymerase chain reaction (qPCR) using TaqMan assays and immunoblotting with specific antibodies. Autophagy activation was determined by measuring the levels of microtubule associated protein 1 light chain 3-II (LC3-II) by immunoblotting and visualization of autophagosomes by transmission electron and confocal microscopy. BME was non-toxic to the OA chondrocytes at the doses employed and suppressed the IL-1β induced expression of inerleukin-6 (IL-6) and matrix metalloprotease-3 (MMP-3), MMP-9 and MMP-13. BME enhanced autophagy in chondrocytes as determined by measuring the levels of LC3-II by immunoblotting and increased number of autophagosomes in BME treated chondrocytes by transmission electron microscopy and confocal microscopy. BME upregulated the expression of several autophagy related genes and increased the autophagy flux in human OA chondrocytes under pathological conditions. Further analysis revealed that BME activated autophagy in chondrocytes via inhibition of mammalian target of rapamycin (mTOR) pathway. Of importance is our finding that BME-mediated suppression of IL-1β induced expression of IL-6, MMP-3, -9, and -13 was autophagy dependent and was abrogated by inhibition of autophagy. The above results show that the Butea monosperma (Lam.) extract has strong potential to activate autophagy and suppress IL-1β induced expression of IL-6 and MMP-3, -9 and -13 in human OA chondrocytes. This study shows that BME or compounds derived from BME can be developed as safe and effective chondroprotective agent(s) that function by activating autophagy to suppress the expression of inflammatory and catabolic factors associated with OA pathogenesis.


Feasibility Study on Phytoremediation of Potentially Toxic Elements (Cd, Ni and Pb) in Aquatic Environments Using Suaeda maritima Plant Species, a Case Study: Khorkhoran International Wetland, Persian Gulf.
The current study was designed to assess the phytoremediation potential of Suaeda maritima has been assessed for cleanup of contaminated sediments with Cd, Ni and Pb. In so doing, totally, 20 sampling sites were selected in the Khorkhoran International Wetland. The contents of elements in sediments, plant organs and water samples were determined using ICP-OES. The mean contents of Cd, Ni and Pb (mg/kg) in the sediment samples were found to be 0.096, 38.1 and 1.78, respectively. Moreover, the mean levels of Cd, Ni and Pb (mg/kg) in root samples of S. maritime were 0.160, 2.72 and 1.22 respectively; whereas, in leaf samples they were found to be 0.157, 3.34 and 2.23 mg/kg, respectively. Furthermore, the mean contents of Cd, Ni and Pb (µg/L) in water samples were 243, 1440 and 3010, respectively. The values of BCF for Cd, BAF for Cd and Pb, and TF for Ni and Pb were higher than 1, which would indicate that S. maritima could possibly be a suitable candidate for the phytostabilization of Cd and the phytoextraction of Ni and Pb.


Predictors and 1-year outcome of major bleeding in patients with non-ST-elevation acute coronary syndromes: insights from the Canadian Acute Coronary Syndrome Registries.
Bleeding is the major noncardiac adverse consequence of treatment in patients with non-ST-elevation (NSTE) acute coronary syndromes (ACS). The aims were to identify predictors of major bleeding in patients with NSTE ACS and to evaluate in-hospital and 1-year outcomes. We evaluated 5842 patients with NSTE-ACS included in the multicenter Canadian ACS Registries. Patients with in-hospital major bleeding (n = 79, 1.4%) were older (75 vs 66 years, P < .001), with higher baseline creatinine (1.17 vs 1.02 mg/dL, P < .002), more diabetes (36% vs 25%, P = .04), hypertension (73% vs 54%, P < .002), previous stroke (20% vs 9%, P < .002), Killip class > or = II (P < .01), and ST-depression (37% vs 18%, P < .001). Antiplatelet/thrombotic therapies did not differ between patients with and without major bleeding. Crude event rates were significantly higher in patients with major bleeding: in-hospital death 19.2% vs 1.5% (P < .001), myocardial infarction (MI) 15.4% vs 3.8% (P < .001), and death or MI 26.9% vs 4.9% (P < .001). One-year event rates were also higher: death 35.9% vs 7.4% (P < .0001), MI 15.6% vs 6.2% (P = .0021), and death or MI 40.6% vs 12.6% (P < .0001). In multivariable analysis, independent predictors for major bleeding were age (OR 1.03, 95% CI 1.03-1.09), hypertension (1.72, 1.0-2.97), previous stroke (1.84, 1.0-3.37), and in-hospital catheterization (1.93, 1.06-3.53) or bypass surgery (2.18, 1.0-4.76). Major bleeding was an independent predictor of 1-year death (OR 3.92, 2.07-7.41) and death or MI (2.51, 1.41-4.48). In patients with NSTE-ACS, major bleeding is associated with increased short- and long-term rates of death and MI, and is predicted by simple clinical characteristics.


MicroRNA-520c-3p inhibits hepatocellular carcinoma cell proliferation and invasion through induction of cell apoptosis by targeting glypican-3.
Glypican-3 (GPC3) is a membrane-associated heparan sulfate proteoglycan involved in regulation of cell proliferation, cell survival, cell migration and differentiation process. MicroRNAs (miRNAs) are single-stranded, non-coding functional RNAs that are important in many biological processes. GPC3 and miRNAs have been found to play essential roles in the development and progression of hepatocellular carcinoma (HCC). However, little information about the relationship between GPC3 and miRNAs is available nowadays. Therefore, this study aims to examine the relationship between GPC3 and miRNAs. Dual-luciferase reporter assay was used to validate the direct target of GPC3. Fluorescence quantitative PCR and Western blotting were used to examined the gene expression at mRNA and protein levels. Cell apoptosis was evaluated by flow cytometric analysis and Annexin V-FITC staining. Invasion of cells was evaluated by Transwell matrigel assay. The results showed that miR-520c-3p could specifically target GPC3 in HCC cells. GPC3 protein levels decreased with unchanged transcription efficiency after miRNA transfection, and there was negative correlation of miR-520c-3p expression in HCC in relate to GPC3 protein levels. Moreover, miR-520c-3p not only induced HCC cell apoptosis, but also inhibited the growth and invasion of the cells. Interestingly, overexpression of GPC3 could effectively reverse apoptosis induced by miR-520c-3p transfection in HCC. Taken together, these results supported that miR-520c-3p may decrease GPC3 protein levels to inhibit proliferation of HCC cells. Therefore, GPC3 could be a new target for genetic diagnosis and treatment of HCC.


Geographical and temporal patterns of incidence and mortality from prostate cancer.
In many countries of the world, prostate cancer is the second most common form of cancer in men, and in the United States it is now in first rank. It is an important public health problem, with > 0.25 million new cases diagnosed worldwide in the year 1985. Whereas earlier large increases in the incidence of prostate cancer were apparent throughout the world, the mortality rate has remained constant in generations of men born since the early years of this century. Most importantly, given that in several countries the increased number of children born after World War II will be in their mid-50s in the early part of the 21st century (at an age when cancer risk is becoming an important consideration), and coupled with the trends in increasing life expectancy, the consequence will be an increase in absolute terms in the number of cases of prostate cancer diagnosed. In the absence of treatment improvements and with prospects for prevention by modification of lifestyle remote within current knowledge, there will also be an increase in the number of deaths from prostate cancer worldwide. The situation would be further augmented by the presence of a temporal trend in risk that is widely reported from many countries and unlikely to be entirely artefact.


Evaluation of miR-302 promoter activity in transgenic mice and pluripotent stem cell lines.
Some miRNAs, including the miR-302 cluster, are critical regulators of the stemness state of embryonic stem cells and cell fate patterning. In this study, we evaluated the activity of the miR-302 core promotor in mice and human pluripotent stem cells, somatic tissue derivatives, and generated transgenic mice expressing EGFP under a miR-302 promoter. The expression of EGFP under the control of the miR-302 promotor was examined in the cell lines and somatic tissues of transgenic mice, transgenic blastocysts, and embryonic stem cells derived from transgenic blastocysts. Our results showed that the miR-302 promoter is highly expressed in the mouse and human pluripotent cells, weakly expressed in the somatic tissue derivatives, is highly expressed in both blastocysts and the first passages of transgenic embryonic stem cells, and lowly expressed in the somatic tissues of transgenic mice. It can be concluded that different temporal and spatial gene expression patterns occur during the embryonic and adult stages of cells in mice.


The Normal and Abnormal Equine Neonatal Musculoskeletal System.
The first weeks of life are critical in many aspects, and the musculoskeletal system is no exception. Being able to stand and nurse within hours of life is necessary for survival. Laxity, flexural deformities, and skeletal immaturity can all make it difficult for neonates to ambulate. The increased vascularity to bones and cartilage mixed with the newly forming immune system also make neonates susceptible to infections that we rarely see in adult animals. This article concentrates on orthopedic conditions seen in the first 2 weeks of life.


Risk factors for noncontact anterior cruciate ligament injury in female high school basketball and handball players: A prospective 3-year cohort study.
The risk factors of noncontact anterior cruciate ligament (ACL) injury remain an enigma. The purpose of this study was to prospectively evaluate the risk factors for noncontact ACL injury in female high school basketball and handball players. We conducted a 3-year prospective cohort study between 2009 and 2014, and it included 317 female high school athletes aged 15 years. At baseline, they underwent detailed examinations for various parameters that were documented during their first year of high school. The parameters assessed were height (cm), weight (kg), general joint laxity (points), navicular drop (mm), anterior laxity of the knee using the KT-1000 Knee Ligament Arthrometer (mm), angle of femoral anteversion (º), knee extensor/flexor muscle strength (Nm/kg), and hip abductor strength (Nm/kg). All ACL injuries that occurred during these 3 years were recorded. Of 317 players, 27 were excluded because they either had a history of ACL injury or could not complete the study. Thirty ACL tears occurred. Three of the ACL injuries were contact injuries, whereas the remaining 27 were noncontact injuries. Greater body weight (95% confidence interval [CI], 1.030-1.174; P = 0.004), a high hip abductor strength (95% CI, 1.462-4.827; P = 0.001), and small femoral anteversion (95% CI, 0.746-0.982; P = 0.027) were found to be risk factors in logistic regression analysis. Greater body weight, a high hip abductor strength, and small femoral anteversion were risk factors for noncontact ACL injury in female high school basketball and handball players.


Effects of psychotropic drugs on methamphetamine-induced behavioral excitation in grouped mice.
When 7 mice injected with methamphetamine (5 mg/kg, s.c.) were placed together into a box with a limited space maintained at 28-30 degrees C, the grouped mice showed extreme behavioral excitation consisting of 2 typical features: fighting behavior with squeak and hyperactive behavior characterized by running and jumping. The effects of various drugs tested, according to the minimal effective doses (mg/kg, p.o.) which inhibited fighting behavior and hyperactive behavior respectively, were: haloperidol, 0.1 (fighting)--0.25 (hyperactivity); clothiapine, 1 - 1; perphenazine, 1 - 1; thiothixene, 1 - 5; chlorpromazine, 5 - 10; diazepam, 5 - 100; reserpine, 100 - 100; phenoxybenzamine, 200 - 200 (no effect). Major tranquilizers except rauwolfia alkaloids completely inhibited both types of excited behavior at low doses, as compared with the taming effects of the drugs on footshock-induced fighting episodes in pairs of mice. On the other hand, minor tranquilizers suppressed only fighting behavior, showing little effect on hyperactive behavior. None of the antidepressants, adrenergic blocking agents and other drugs showed antagonism to both types of excited behavior. The results indicate that this method may be useful for the evaluation of psychotropic drug effects in grouped mice.


Further studies on the characterization of cylindrin and torin, two extrinsic proteins of the erythrocyte membrane.
The subunit composition of the high molecular weight proteins cylindrin and torin from human erythrocyte ghosts has been studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis on 3 to 30% "Gradipore' polyacrylamide gradient slab gels. Torin has been shown to be a multimer of a single polypeptide of approx. Mr 20 000. Cylindrin appears to contain five polypeptides, three of which predominate, in the Mr range 22 000 to 25 000. The isoelectric points (pI) of cylindrin and torin have been determined as 4.6 and 4.8, respectively. The molecular properties of cylindrin and torin are discussed in relation to the previous studies by the authors and others on these proteins.


Molecular and Cellular Advances in Endometriosis Research: Paving the Way for Future Directions.
Molecular and cellular research in the field of endometriosis is moving forward in giant steps [...].


Rate of relapse following treatment for localized prostate cancer: a critical analysis of retrospective reports.
Controversy exists over the optimal treatment for patients with clinically localized prostate cancer. Almost all of the treatment results are from non-randomized trials and interseries comparison is difficult since the apparent success of a treatment, as judged by the actuarial freedom from relapse and survival data, depends on patient selection criteria and post-treatment evaluation, in addition to the efficacy of the therapeutic intervention. In this report the calculation of a hazard function is used to estimate and compare the rate of relapse for the different treatments. Clinical reports from major surgery and radiation oncology treatment institutions were analyzed. The actuarial recurrence data were used to calculate the annual rate of recurrence within each series. For all but the lowest volume tumors, patients continue to be at risk of relapse for as long as these series have been followed. Despite the heterogeneity of patient populations, the recurrence rates by stage are similar for patients treated with surgery or irradiation. This result is consistent with pathologic data from prostatectomy specimens which indicate that for lesions > 12 cm3 (approx. 3 cm in diameter) there is high likelihood of extraprostatic disease. Treatment outcome for patients with localized prostate cancer may be more dependent on the inherent tumor biology than the particular type of treatment. Accordingly, the expectation and recommendation of a treatment must take into consideration the continued risk of relapse with either radiation therapy or surgery. There are, as yet, insufficient data regarding the impact of screening and earlier diagnosis on the curability of patients with localized prostate cancer.


Enterodiol and enterolactone, two major diet-derived polyphenol metabolites have different impact on ERalpha transcriptional activation in human breast cancer cells.
Lignans are plant compounds metabolized in the mammalian gut to produce the estrogenic enterolignans, enterodiol (ED) and enterolactone (EL). Because estrogens have been linked to breast cancer etiology, enterolignans could affect breast cancer risk, but to our knowledge, the mechanisms by which they exert their estrogenic and/or anti-estrogenic effects in humans are still unclear. To better understand how estrogenic compounds from the food, such as the enterolignans, might influence breast cancer progression and their mechanisms to interfere with human estrogen receptor (ER) signalling in hormone-dependant diseases, we examined and compared the ability of ED, EL and 17beta-estradiol (E2) to induce the transactivation of ERalpha and ERbeta, to modulate ERalpha target genes, to exert either growth stimulatory or anti-proliferative effects and finally to modulate MCF-7 cell migration by acting on matrix metalloproteases (MMP)-2 and -9, at concentrations that are achievable through a lignan-rich diet. This study indicates that enterolignans show distinct properties for transactivation of ERalpha and ERbeta. ED, as E2, induces ERalpha transcriptional activation through transactivation functions AF-1 and AF-2, while EL is less efficient in inducing AF-1, acting predominantly through AF-2. Furthermore, ED and EL modulate ERalpha mRNA and protein contents as well as MCF-7 cell proliferation and secreted MMP activities in a different way. Enterolignans are compounds of wide interest nowadays and our results help to unveil their mechanisms of action on ER, emphasizing the fact that the dietary load in lignans could be of importance in the balance between being risk or chemopreventive factors for breast cancer and women's health.


A Case Report of Dendritic Fibromyxolipoma in the Back of Pediatric Patient.
To investigate the pathologic features and differential diagnosis of a pediatric dendritic fibromyxolipoma (DFML). We collected one case of dendritic fibromyxolipoma in the back of a pediatric patient (6 years old, female) diagnosed at the First Affiliated Hospital of Dalian Medical University. The case of DFML was studied by H&E and MaxVision immunohistochemical staining. The clinical and pathological features were analyzed with review of the literatures RESULTS: The tumor was round or ovoid in shape and appeared in the surface of back. Microscopically, the tumor was mainly consisted of stellate to spindle cells, rope-like collagen fibers and varying amount of mature lipocytes embedded in myxoid stroma. Immunohistochemical staining showed that spindle cells and stellate cells were positive for vimentin, CD34 and BCL-2. DFML is a rare special variant of lipoma. It is benign, and it seems to be misdiagnosed as myxoid liposarcoma prior to the admission to our hospital. The pathological histomorphology and immunohistochemistry phenotypes are helpful to the diagnosis and differential diagnosis. Given that DFML mainly occurs in the elderly, this case is rare and worthy to be reported since it occurs in children.


Improving viral filtration capacity in biomanufacturing processes using aggregate binding properties of polyamide-6,6.
Virus retention filtration is a common step in modern biopharmaceutical manufacturing as it enables efficient removal of potential adventitious and endogenous viruses via size exclusion. Modern parvovirus retention filters have significantly improved fluxes and parvovirus retention in comparison to earlier versions of these filters. However, these filters may be more susceptible to premature fouling and require more effort for process optimization. Here, we demonstrate that polyamide-6,6 (nylon-6,6) membranes when used as prefilters can increase the capacity of these Parvovirus retentive filters that are less susceptible to premature fouling. We found that the mechanism of polyamide-mediated filtration improvement can be explained by the binding of monoclonal antibody (mAb) aggregates with a diameter of 20-100 nm, and we show that this mechanism is shared by other types of adsorptive prefilters. Finally, by the combination of mobile phase screening, additive spiking, and molecular dynamics simulations, we show that polyamide-6,6 removes mAb aggregates through hydrophobic interactions making its design space potentially complementary to other available prefilters. Our studies support the aggregate-mediated mechanism of flux decay during viral filtration and suggest that polyamide-6,6 could be considered as an alternative cost-effective option to extend the capacity of viral filters.


Current evidence on the cytotoxic T-lymphocyte antigen 4 + 49G > A polymorphism and digestive system cancer risks: a meta-analysis involving 11,923 subjects.
Cytotoxic T-lymphocyte antigen (CTLA-4) plays an important role in downregulating T cell activation and proliferation. The CTLA-4 + 49G > A polymorphism is one of the most commonly studied polymorphisms in this gene due to its association with many cancer types, but the association between CTLA-4 + 49G > A polymorphism and digestive system cancer risks remain inconclusive. An updated meta-analysis based on 17 independent case-control studies consisting of 5176 cancer patients and 6747 controls was performed to address this association. Overall, there was no statistically increased risk of digestive system cancers in every genetic comparison. In subgroup analysis, this polymorphism was significantly linked to higher risks for pancreatic cancer (GG vs. AA, OR = 1.976, 95% CI = 1.496-2.611; GA vs. AA, OR = 1.433, 95% CI = 1.093-1.879; GG/GA vs. AA, OR = 1.668, 95% CI = 1.286-2.164; GG vs. GA/AA, OR = 1.502, 95% CI = 1.098-2.054; G vs. A, OR = 1.394, 95% CI = 1.098-1.770). We also observed increased susceptibility of hepatocellular cell carcinoma in homozygote comparison (OR = 1.433, 95% CI = 1.100-1.866) and dominant model (OR = 1.360, 95% CI = 1.059-1.746). According to the source of controls, significant effects were only observed in hospital-based studies (GA/AA vs. GG, OR = 1.257, 95% CI = 1.129-1.399). In the stratified analysis by ethnicity, no significantly increased risks were found in either Asian or Caucasian. Our findings suggest that the CTLA-4 + 49G > A polymorphism may be associated with the risk of pancreatic cancer and hepatocellular cell carcinoma.


Central fatigue contributes to the greater reductions in explosive than maximal strength with high-intensity fatigue.
The study aimed to assess the influence of fatigue induced by repeated high-force explosive contractions on explosive and maximal isometric strength of the human knee extensors and to examine the neural and contractile mechanisms for the expected decrement. Eleven healthy untrained males completed 10 sets of voluntary maximal explosive contractions (five times 3 s, interspersed with 2 s rest). Sets were separated by 5 s, during which supramaximal twitch and octet contractions [eight pulses at 300 Hz that elicit the contractile peak rate of force development (pRFD)] were evoked. Explosive force, at specific time points, and pRFD were assessed for voluntary and evoked efforts, expressed in absolute terms and normalized to maximal/peak force. Maximal voluntary contraction force (MVCF) and peak evoked forces were also determined. Surface EMG amplitude was measured from three superficial agonists and normalized to maximal compound action potential area. By set 10, explosive force (47-52%, P < 0.001) and MVCF (42%, P < 0.001) had declined markedly. Explosive force declined more rapidly than MVCF, with lower normalized explosive force at 50 ms (29%, P = 0.038) that resulted in reduced normalized explosive force from 0 to 150 ms (11-29%, P ≤ 0.038). Neural efficacy declined by 34%, whilst there was a 15-28% reduction in quadriceps EMG amplitude during voluntary efforts (all P ≤ 0.03). There was demonstrable contractile fatigue (pRFD: octet, 27%; twitch, 66%; both P < 0.001). Fatigue reduced normalized pRFD for the twitch (21%, P = 0.001) but not the octet (P = 0.803). Fatigue exerted a more rapid and pronounced effect on explosive force than on MVCF, particularly during the initial 50 ms of contraction, which may explain the greater incidence of injuries associated with fatigue. Both neural and contractile fatigue mechanisms appeared to contribute to impaired explosive voluntary performance.


[Concentrated autologous tissue glue].
Since 1980 an autologous tissue glue has been used for microsurgery and plastic surgery at the University of Graz, Clinic for Oto-rhino-laryngology. This tissue glue is made from the patient's own blood plasma. The advantages are: No danger of an immune reaction or infection by virus hepatitis, no storage problems, easy to synthesize, few expenses. Because of the low concentration of fibrinogen, the stability at the beginning of gluing, compared to the other tissue glues, was less. We succeeded in producing a new autologous tissue glue with a highly concentrated amount of fibrinogen, which provided a better stability at the beginning of gluing. According to application, the content of fibrinogen can be varied. The adhesiveness of the concentrated autologous tissue glue was tested and compared to another kind of homologous tissue glue.


Identification of the three non-identical subunits constituting human deoxyribonuclease II.
We purified DNase II from human liver to apparent homogeneity. The N-terminal amino acid sequences of each of three components constituting the purified mature enzyme were then separately determined by automatic Edman degradation. A combination of this chemical information and the previously reported nucleotide sequence of the cDNA encoding human DNase II [Yasuda et al. (1998) J. Biol. Chem. 273, 2610-2626] allowed detailed elucidation of the enzyme's subunit structure: human DNase II was composed of three non-identical subunits, a propeptide, proprotein and mature protein, following a signal peptide. Expression analysis of a series of deletion mutants derived from the cDNA of DNase II in COS-7 cells suggested that although a single large precursor protein may not be necessary for proteolytic maturation, the propeptide region L17-Q46 may play an essential role in generating the active form of the enzyme.


Intraluminal pressure variations in the isthmus of the porcine oviduct after intrauterine insemination with saline, oestrogen solution or boar seminal plasma.
The purpose of this study was to investigate if an intrauterine deposition of saline, boar seminal plasma, or an oestrogen solution containing 11.5 micrograms oestrogens affected the intraluminal pressure in the isthmus of the oviduct in unrestrained gilts. In order to monitor variations in intraluminal pressure, 2 ultra-miniature pressure sensors, located at 2 different points along the same isthmus were used. After an initial recording period, either saline, boar seminal plasma, or the oestrogen solution was deposited directly into the uterus. Intraluminal pressure recordings were conducted up to 6 h after insemination, and blood samples, for monitoring oestradiol-17 beta and 15-ketodihydroprostaglandin F2 alpha levels, were collected. None of the inseminates had any consistent effect on the intraluminal pressure in the porcine oviduct. After deposition of the oestrogen solution, increases in the circulating levels of both oestradiol-17 beta and 15-ketodihydroprostaglandin F2 alpha were seen.


Priming of the respiratory burst of neutrophils by diacylglycerol. Independence from activation or translocation of protein kinase C.
Preincubation of neutrophils with certain agonists may "prime" the cells to cause increased responses to a second stimulus ("primed stimulation"). We used two approaches to examine the role of protein kinase C (Ca2+/phospholipid-dependent enzyme) in priming and stimulation by 1-oleoyl-2-acetylglycerol (OAG), phorbol 12-myristate 13-acetate (PMA), and N-formyl-Met-Leu-Phe (fMLP): inhibition of protein kinase C by 1-(5-isoquinolinesulfonyl)-piperazine (C-I) and measurement of protein kinase C translocation induced by priming and stimulatory concentrations of OAG. C-I had little effect on stimulation or primed stimulation by fMLP, suggesting that fMLP invokes events independent of protein kinase C. C-I equally inhibited stimulation and primed stimulation by PMA. Direct stimulation by OAG was inhibited, but priming and primed stimulation by OAG was unaltered by C-I. OAG concentrations greater than or equal to 100 microM caused translocation of protein kinase C, in correlation with direct stimulation of the respiratory burst. Lower OAG concentrations (10-30 microM) primed to stimulation by fMLP and, conversely, stimulated neutrophils primed with fMLP, yet did not cause translocation of protein kinase C. The data are compatible with previous assumptions that PMA and OAG directly stimulate polymorphonuclear neutrophil leukocytes by translocation and activation of protein kinase C. However, priming and primed stimulation by OAG apparently invoke distinct transduction mechanisms other than protein kinase C translocation.


A clinical study comparing marginal and occlusal accuracy of crowns fabricated from double-arch and complete-arch impressions.
The double-arch impression technique is widely used in the provision of laboratory fabricated restorations. However, there is little clinical evidence to support or refute its use. The aim of this prospective clinical study was to evaluate the occlusion and marginal fit of posterior full crowns made from double-arch impressions, and to compare these to control crowns fabricated from conventional complete-arch impressions. Ten patients requiring single posterior ceramo-metal full crowns had both double-arch and complete-arch impressions taken of the prepared tooth. Two crowns were fabricated for each tooth, using the casts made from both impression methods. Both crowns were tried in and the occlusion and margins evaluated. The results were composed using nonparametric statistical analysis with the probability level for significance at alpha=0.05. The crowns fabricated from the double-arch impression were found to be more accurate in closure to the intercuspal position and had fewer interferences in lateral excursions. There were no significant differences between the two crown groups regarding protrusive interferences and margin quality. Crowns fabricated from the double-arch impressions were equivalent in marginal accuracy and superior occlusally to crowns fabricated from the complete-arch impressions.


[Utilization rates and turnover times as indicators of OR workflow efficiency].
In many hospitals operating room (OR) utilization rates and turnover times (the time from the end of the previous surgical procedure to the beginning of the next) are used as indicators of OR workflow inefficiency. However, there have been no detailed studies to determine whether these indicators really provide an adequate picture of avoidable wasting of time in the OR. All relevant OR processes in a busy surgical suite with nine ORs were studied in detail over an 8-week period. Productive OR processes, and also reasons for unused times, were recorded by independent observers at 5-minute intervals; they were able to code for 10 different productive activities and 20 different reasons for unused time. Unused time in the OR, the OR utilization rate and the average perioperative turnover times were calculated for each day and a correlation analysis was performed. In all, 3,501 OR hours and 790 surgical cases were studied. Productive processes accounted for 85.7% of the total OR time; the unused times were times with no scheduled cases (7.7%) and waiting times that arose for many different reasons (6.6%). Correlation analysis showed that there was no close correlation between waiting time and OR utilization (Spearman's r(s) 0.104 and r(s) 0.233). The correlations between total unused time (r(s) 0.718 and r(s) 0.745) and time with no scheduled cases (r(s) 0.706 and r(s) 0.620) and utilization were more robust, but for any given OR utilization rate the range of corresponding unused time or time without scheduled cases per day was considerable. The correlation between waiting time and perioperative turnover times was negligible (r(s) 0.185 and r(s) 0.175). When different definitions of utilization rate or perioperative turnover were used the results obtained were virtually identical. Utilization rate and perioperative turnover time cannot be used as indicators of OR workflow efficiency, since they cannot identify the days during which avoidable waiting times occur. If the aim is to identify underused OR time and factors that hamper workflow efficiency, waiting times and times without scheduled cases need to be recorded directly and separately.


Evaluating the performance of three different cervical cancer screening modalities in a large prospective population-based cohort.
This study aimed to evaluate three different patterns of cervical cancer screening strategies for detection of cervical diseases. In total, 10,186 women aged 21-70 years attending cervical screening program were recruited and cotested by human papillomavirus (HPV) assays and cytology. Three-year histological follow-up data was recorded on women with abnormal screening results, and six clinically common screening algorithms were evaluated. Significantly better protection against cervical intraepithelial neoplasia 2 or worse (CIN2+) at three-year follow-up was associated with a negative HPV result than by normal cytology at baseline. HPV screening was more sensitive and less specific than cytology screening. Moreover, HPV screening with HPV16/18 and reflex cytology (atypical squamous cells of undetermined significance [ASCUS] threshold) showed a similar sensitivity (94.6% vs. 98.2%, p = 0.125) and a superior specificity as compared to cotesting reflex HPV16/18 and cytology (ASCUS threshold) for CIN2+ (95.8% vs. 95.1%, p < 0.0001), achieving a colposcopy referral rate of 5.4%, and consuming 4.8 colposcopies and 4.4 cytology tests to find one CIN2+. HPV screening with triage of HPV-positive women by HPV16/18 genotyping and cytology provided a good equilibrium between screening effectiveness, the number of cytology tests required, and referral rates; HPV testing was similar in sensitivity to cotesting and safer than cytology, thus especially suitable for large population-based screening programs.


An examination of multiple factors affecting community structure in an aquatic amphibian community.
The potential effects of multiple factors structuring certain larval amphibian communities were studied using a pen experiment in a natural pond. Potential factors (predation and competition from other species) were allowed to act in a stepwise fashion such that their relative importance could be evaluated. Based on a previous study, it was hypothesized that predation by Ambystoma salamander larvae on other larval amphibian species would be the most important factor. Survival of Ambystoma jeffersonianum salamander larvae and Rana sylvatica tadpoles was significantly depressed only by Ambystoma opacum predation. Survival of Ambystoma maculatum salamander larvae was significantly greater in the absence of both A. opacum and A. jeffersonianum predators. The virtual elimination of Hyla chrysoscelis larvae in all treatments also can be largely attributed to Ambystoma predation. Thus, Ambystoma predation was the dominant factor determining larval survival of four amphibian prey species in the experimental communities.


Magnetoencephalography recordings reveal the spatiotemporal dynamics of recognition memory for complex versus simple auditory sequences.
Auditory recognition is a crucial cognitive process that relies on the organization of single elements over time. However, little is known about the spatiotemporal dynamics underlying the conscious recognition of auditory sequences varying in complexity. To study this, we asked 71 participants to learn and recognize simple tonal musical sequences and matched complex atonal sequences while their brain activity was recorded using magnetoencephalography (MEG). Results reveal qualitative changes in neural activity dependent on stimulus complexity: recognition of tonal sequences engages hippocampal and cingulate areas, whereas recognition of atonal sequences mainly activates the auditory processing network. Our findings reveal the involvement of a cortico-subcortical brain network for auditory recognition and support the idea that stimulus complexity qualitatively alters the neural pathways of recognition memory.


Iron oxide nanoparticles as magnetic relaxation switching (MRSw) sensors: Current applications in nanomedicine.
Since pioneering work in the early 60s on the development of enzyme electrodes the field of sensors has evolved to different sophisticated technological platforms. Still, for biomedical applications, there are key requirements to meet in order to get fast, low-cost, real-time data acquisition, multiplexed and automatic biosensors. Nano-based sensors are one of the most promising healthcare applications of nanotechnology, and prone to be one of the first to become a reality. From all nanosensors strategies developed, Magnetic Relaxation Switches (MRSw) assays combine several features which are attractive for nanomedical applications such as safe biocompatibility of magnetic nanoparticles, increased sensitivity/specificity measurements, possibility to detect analytes in opaque samples (unresponsive to light-based interferences) and the use of homogeneous setting assay. This review aims at presenting the ongoing progress of MRSw technology and its most important applications in clinical medicine.


Non-contrast-enhanced MR imaging of the renal arteries.
In this article, we focus on non-contrast magnetic resonance angiography techniques for evaluating renal artery imaging. Time-of-flight, phase contrast, steady-state free procession, and arterial spin labeling are discussed.


Variations in traffic-related polycyclic aromatic hydrocarbons in PM2.5 in Kanazawa, Japan, after the implementation of a new vehicle emission regulation.
A three-year sampling campaign was conducted at a roadside air pollution monitoring station in the urban area of Kanazawa, Japan. Due to a new emission regulation, PAHs levels decreased over the sampling campaign, exhibiting values of 706 ± 413 pg/m3 in 2017, 559 ± 384 pg/m3 in 2018, and 473 ± 234 pg/m3 in 2019. In each year, similar seasonal variations in PAHs levels were observed, with higher levels observed in winter and lower levels in summer. Among the PAHs isomer ratios, we observed that the ratio of benzo[b]fluoranthene (BbF) and benzo[k]fluoranthene (BkF), [BbF]/([BbF] + [BkF]), and the ratio of indeno[1,2,3-cd]pyrene (IDP) and benzo[ghi]perylene (BgPe), [IDP]/([BgPe] + [IDP]), showed stability over the sampling campaign and were less affected by the new emission regulation, seasonal variations, and regional characteristics. When using the combined ratio ranges of 0.66 - 0.80 ([BbF]/([BbF] + [BkF]) and 0.26-0.49 ([IDP]/([BgPe] + [IDP]), traffic emissions were clearly distinguished from other PAHs emission sources. Principal component analysis (PCA) and positive matrix factorization (PMF) were also performed to further analyse the characteristics of traffic-related PAHs. Overall, this study affirmed the effectiveness of the new emission regulation in the reduction of PAHs emissions and provided a combined range for identifying PAHs traffic emission sources.


[Interaction of heterosensory afferent impulses in the neurons of the thalamic associative nuclei].
The interaction of visual, auditory and cutaneous afferent signals on neurons of pulvinar (Pulv), posterior lateral (LP) and mediodorsal (MD) thalamic nuclei was studied in acute experiments on cats anaesthetized with a mixture of nembutal and chloralorse. The first (more frequent) type of the interaction was characterized by inhibition or inhibition followed by facilitation of the response; the second--by facilitation or facilitation followed by inhibition of neuronal activity. The capacity for interaction in MD neurons was less expressed than in cells of the Pulv-LP complex (P less than 0.05). The interaction could be better seen in three-sensory elements. Certain peculiarities were found in the interaction of afferent signals coming from different peripheral receptive areas in neurons of both structures.


The inhibitory kinetics of vitamins B9, C, E, and D3 on bovine xanthine oxidase: Gout treatment.
Over-consumption of foods high in purines like seafood, red meat, and alcoholic beverages leads to hyperuricemia causing gout attacks. Xanthine oxidase was reported responsible for the overproduction of uric acid. We intend to test in silico and in vitro, the inhibition effect of four vitamins against bovine milk xanthine oxidase (BXO). We performed Molecular docking with GOLD v4.0, and the biological activity prediction with the PASS server. The best-selected vitamins were chosen based on their best PLPchem score. The BXO constant Km and Vmax were determined in vitro, and then the vitamins were tested for their inhibition effect to BXO. Furthermore, the inhibition constant Ki of each inhibitor were determined using Dixon method, the vitamins chosen were vitamin E, vitamin B9, vitamin D3, and vitamin C. The in silico results show that the tested vitamins were the best inhibitors model with PLPchem scores up to 70 comparing to the control. The in vitro results show that BXO have a Km value of 163.55 μM with Vmax of 37 U, vitamins B9, E, C, and D3 were potent inhibitors to BXO with an IC50 of 34.10 ± 0.21, 36.68 ± 1.50, 39.01 ± 0.02, and 100.28 ± 0.33 μM, respectively comparing to the control (32.03 ± 0.73 μM). The kinetic study shows that all tested vitamins were Non-competitive inhibitors, the Ki values were 15 ± 1.76 μM, 29 ± 1.06 μM, 12 ± 1.41 μM, and 20 ± 0.71 μM, for respectively vitamins B9, E, C, and D3. The obtained results promise an excellent strategy using vitamins to enhance immunity, treat hyperuricemia, and minimize the usual drug side effects.