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Molecular dynamics simulations were performed in aqueous solution to elucidate an atomistic level picture of complex formation between cucurbit[7]uril (CB7) and three standard aromatic amino acids: tyrosine, tryptophan, and phenylalanine. It was found that all three amino acids formed stable host–guest complexes with CB7, in which the side chain was included inside the hydrophobic cavity and the ammonium and carboxylate groups were excluded. The major forces driving complexation, as calculated from the MM-PBSA method, were the electrostatic and van der Waal interactions. To better understand the effect of pH and the roles of the ammonium and carboxylate groups in the complexation process, simulations were performed for phenylalanine considering different protonation state (at low and high pH) as well as for the deaminated and decarboxylated forms of phenylalanine. The results showed that, compared to phenylalanine at neutral pH (exists as zwitterion), low pH resulted in an increased complex stability for the cationic form, deamination and high pH reduced the stability, while decarboxylation did not result in a significant change. Results from quantum-chemical calculations correlated well with the simulation data.

During storage and thermal treatment meat suffers a number of biochemical and physical-chemical changes in the substrate protein, changes that take place with varying intensity depending on the method of preservation utilized and temperature of thermal treatment applied. Application of different treatments aimed to influence the proteolytic activity as is the case of enzymatic tenderization of beef.Improving the meat tenderness with proteolytic enzymes is promising, but current legislation restricting the use of proteolytic enzymes from bacterial origin and recommended tenderizers salts containing papain, ficin and bromelain. Recent research revealed that meat marinating before grilling results in a reduction of heterocyclic amine content after thermal treatment. Also, the addition of fruit pulp, garlic or other spices contributes to decreased production of heterocyclic amines because of their antioxidant activity. In the present study was aimed influence of exogenous proteolytic enzymes on adult beef tenderness. To increase the tenderness of adult beef were used exogenous enzymes preparations (papain and bromelain) and natural sources of enzymes using pineapple and papaya fruit. It was intended to establish the correlation between enzymatic activity of enzymes used in the study, the processing technology and changes in the physical-chemical and biochemical characteristics that occur during storage in refrigerated conditions (evolution of the rigidity index and water holding capacity, cooking losses and cooking yield of the samples injected/marinated with enzymes).

Objectives Multiple aspects of the metabolic syndrome (MetS) remain problematic. Here we assess the association between epidemic obesity and the other MetS traits, and MetS utility for cardiovascular disease (CVD) risk assessment. Methods Italian population-based Progetto CUORE data were used: 17 252 women and men ages 35–69 years, baseline 1984–1993, mean follow-up of 10 years, for nonfatal plus fatal CVD events. NCEP-ATP III criteria defined MetS. Results Epidemic obesity was strongly related to epidemic rates of the four other MetS traits. Only four of 16 possible MetS trait combinations were common; their CVD hazard ratios ranged from 1.21 to 1.70. In multivariate analyses MetS was no better than the sum of its parts in predicting CVD, important information was lost due to omission of non-HDL-C and smoking, and from considering MetS traits as yes/no variables. CVD risk prediction by MetS was less strong for men and no stronger for women than by classical risk factors (blood pressure, diabetes, serum cholesterol, smoking, overweight/obesity). Conclusions These findings are concordant with the inference that epidemic obesity importantly influences epidemic occurrence of the other MetS traits; they also indicate that use of MetS for CVD risk assessment has limitations and needs critical reconsideration.

objective: To analyse the role of recombinant HIV-1 protein p17 in the modulation of cell activity. Methods: Peripheral blood mononuclear cells (PBMC) obtained from healthy donors were cultured in the presence or absence of p17 with mitogens such as phytohaemagglutinin or interleukin-2 and their response assayed by cell proliferation. Cross-linking experiments were employed to investigate the presence of a binding between p17 and factor(s) present in human serum. An immunoenzymatic assay for p24 antigen detection was used to analyse the effect of the addition of exogenous p17 to cultures of PBMC infected with HIV-1 in vitro. Results: Purified recombinant p17 protein at a concentration of 0.25 μg/ml significantly increased the proliferation of preactivated PBMC obtained from healthy donors. This effect was obtained by binding p17 to factor(s) present in human serum and observed on both CD4 + and CD8 + T cells. Recombinant p17 also induced an increased rate of HIV-1 replication, probably due to enhanced T-cell proliferation. The activity of p17 protein was inhibited by anti-p17 antibodies generated by injecting recombinant p17 in rabbits, but not by human antibodies generated during the natural course of HIV infection. Conclusion: Characterization of the human factor(s) and identification of the interacting p17 epitope(s) will improve our understanding of the mechanisms used by HIV to efficiently replicate in our organisms.

The relationship between the number of circulating CD4+ T cells and the presence of particular CD8+ T cell subsets was analyzed by flow cytometry on PBL from asymptomatic HIV-1-infected patients whose specimens were collected every 2 mo for a total period of 32 mo. Only slight variations were detected in the absolute number of lymphocytes and percentage of CD3+ lymphocytes, whereas both CD4+ and CD8+ T cell subsets showed wide intrapatient variation. Variations in the number of CD8+CD28+ cells paralleled those of the CD4+ T cell subset in each patient tested, while the presence of CD8+CD28- T cells correlated inversely with CD4+ and CD8+CD28+ T cells. These data show that changes in the number of circulating CD4+-and CD8+CD28+ T cells are strongly related to the presence of CD8+CD28- T cells in these patients. Insight into the significance of CD8+CD28- T cell expansion will allow us to understand the mechanisms and significance of the HIV-1- driven change in CD4+CD8+ T cell homeostasis and the basic immunopathology of HIV disease.

The expression of activation antigens, namely CD25, CD69, CD71, and HLA-DR on T cells from 15 healthy individuals stimulated with different mitogens and specific antigens was evaluated by immunofluorescence assay and flow cytometric analysis and compared with cell proliferation as a function of [3H]thymidine incorporation. CD69 was the earliest expressed antigen on stimulated cells, while HLA-DR was the latest. Regardless of the stimulus used, lymphocytes expressing CD25 and CD71 were always more numerous than cells expressing CD69 and HLA-DR. Variations in the proportion of CD4+ and CD8+ T cells expressing each activation marker were observed with different antigenic stimuli. The expression of each activation marker showed overall agreement with the [3H]thymidine incorporation assay in discriminating between positive and negative immune response. However, no correlation was observed between the percentage of CD25-, CD69-, CD71-, and HLA-DR-positive T cells and the amount of [3H]thymidine incorporation. Moreover, low doses of mitogens and antigens as well as short time of stimulation were sufficient to induce T cells to express activation antigens but not to proliferate. Our data show that results obtained by flow cytometry and [3H]thymidine incorporation may differ qualitatively, at least under certain conditions; this suggests that the 2 assays are complementary, and when combined, may gives a clearer understanding of events leading to efficient cell-mediated immune response.

BACKGROUND The Centers for Medicare & Medicaid Services publicly reports risk-standardized mortality rates (RSMRs) within 30-days of admission and, in 2013, risk-standardized unplanned readmission rates (RSRRs) within 30-days of discharge for patients hospitalized with acute myocardial infarction (AMI), heart failure (HF), and pneumonia. Current publicly reported data do not focus on variation in national results or annual changes. OBJECTIVE Describe U.S. hospital performance on AMI, HF, and pneumonia mortality and updated readmission measures to provide perspective on national performance variation. DESIGN To identify recent changes and variation in national hospital-level mortality and readmission for AMI, HF, and pneumonia, we performed cross-sectional panel analyses of national hospital performance on publicly reported measures. PARTICIPANTS Fee-for-service Medicare and Veterans Health Administration beneficiaries, 65 years or older, hospitalized with principal discharge diagnoses of AMI, HF, or pneumonia between July 2009 and June 2012. RSMRs/RSRRs were calculated using hierarchical logistic models risk-adjusted for age, sex, comorbidities, and patients’ clustering among hospitals. Results Median (range) RSMRs for AMI, HF, and pneumonia were 15.1% (9.4–21.0%), 11.3% (6.4–17.9%), and 11.4% (6.5–24.5%), respectively. Median (range) RSRRs for AMI, HF, and pneumonia were 18.2% (14.4–24.3%), 22.9% (17.1–30.7%), and 17.5% (13.6–24.0%), respectively. Median RSMRs declined for AMI (15.5% in 2009–2010, 15.4% in 2010–2011, 14.7% in 2011–2012) and remained similar for HF (11.5% in 2009–2010, 11.9% in 2010–2011, 11.7% in 2011–2012) and pneumonia (11.8% in 2009–2010, 11.9% in 2010–2011, 11.6% in 2011–2012). Median hospital-level RSRRs declined: AMI (18.5% in 2009–2010, 18.5% in 2010–2011, 17.7% in 2011–2012), HF (23.3% in 2009–2010, 23.1% in 2010–2011, 22.5% in 2011–2012), and pneumonia (17.7% in 2009–2010, 17.6% in 2010–2011, 17.3% in 2011–2012). Conclusions We report the first national unplanned readmission results demonstrating declining rates for all three conditions between 2009–2012. Simultaneously, AMI mortality continued to decline, pneumonia mortality was stable, and HF mortality experienced a small increase. Electronic supplementary material The online version of this article (doi:10.1007/s11606-014-2862-5) contains supplementary material, which is available to authorized users.

Background Delays in treatment time are commonplace for patients with ST-segment elevation acute myocardial infarction who must be transferred to another hospital for percutaneous coronary intervention. Experts have recommended that door-in to door-out (DIDO) time (ie, time from arrival at the first hospital to transfer from that hospital to the percutaneous coronary intervention hospital) should not exceed 30 minutes. We sought to describe national performance in DIDO time using a new measure developed by the Centers for Medicare & Medicaid Services. Methods We report national median DIDO time and examine associations with patient characteristics (age, sex, race, contraindication to fibrinolytic therapy, and arrival time) and hospital characteristics (number of beds, geographic region, location [rural or urban], and number of cases reported) using a mixed effects multivariable model. Results Among 13 776 included patients from 1034 hospitals, only 1343 (9.7%) had a DIDO time within 30 minutes, and DIDO exceeded 90 minutes for 4267 patients (31.0%). Mean estimated times (95% CI) to transfer based on multivariable analysis were 8.9 (5.6-12.2) minutes longer for women, 9.1 (2.7-16.0) minutes longer for African Americans, 6.9 (1.6-11.9) minutes longer for patients with contraindication to fibrinolytic therapy, shorter for all age categories (except >75 years) relative to the category of 18 to 35 years, 15.3 (7.3-23.5) minutes longer for rural hospitals, and 14.4 (6.6-21.3) minutes longer for hospitals with 9 or fewer transfers vs 15 or more in 2009 (all P Conclusion Among patients presenting to emergency departments and requiring transfer to another facility for percutaneous coronary intervention, the DIDO time rarely met the recommended 30 minutes.

While the Azov and Black seas are subject to anthropogenic pollution to a much higher extent than any other seas, this has been little studied with only a few critical reviews of contaminant fluxes to these seas. Riverine fluxes of the organochlorine pesticides (OCPs) hexachlorocyclohexane (HCH) and DDT to the Azov and Black seas from the former Soviet Union and Russian Federation were thus reviewed for 1986 to 1996. The review was based on official data and data obtained by independent specialists. The amount of HCH used, and the intensity of usage, in these river catchments decreased during the review period. Concurrently, OCP concentrations in the rivers and their fluxes also decreased according to both official and independent data. A comparison of the official and the independent data sets for 1988 revealed significant differences, reflecting the need for more rigorous sampling and analytical protocols for both data sets. According to the OGSNK/GSN data, the flux rates of the five largest rivers were ranked (from largest to smallest) as follows: Don > Dnestr > Danube > Kuban > Dnepro (alpha-HCH); Danube > Don > Dnestr > Dnepro > Kuban (gamma-HCH); Dnestr > Danube > Don > Dnepro-Kuban (DDT+DDE). For rivers with lower annual riverine discharges, the DDT fluxes were surprisingly high (0.43 to 1.49 tonnes a(-1)). According to independent data for 1988 the rankings of the rivers was: Danube > Don > Dnepro > Dnestr > Kuban (alpha-HCH); Danube > Don > Dnestr > Dnepro > Kuban (gamma-HCH); Danube > Dnepro > Dnestr > Don > Kuban (DDT); Danube > Dnepro > Don > Kuban > Dnestr (DDE). The DDT flux estimates for small rivers derived from independent data were 19 to 46 times lower than those calculated using OGSNK/GSN data. According to the independent data, the total riverine OCP transport from the Russian Federation into the Azov Sea from 1988 to 1996 was 1.288 tonnes of gamma-HCH+alpha-HCH and 1.693 tonnes of DDT+DDE while for the Black Sea they were 3.830 tonnes and 5.116 tonnes for gamma-HCH+alpha-HCH and DDT+DDE, respectively.

The synergy between glycopeptides and beta-lactams was studied using different techniques such as broth macrodilution, killing curves and agar dilution combined with agar diffusion. Two glycopeptide-resistant enterococci isolated from different clinical samples were used. Results showed different effects with significant changes in MICs. Antibacterial activity was related to the concentration of glycopeptide and beta-lactam for Enterococcus faecalis 8253, while for Enterococcus faecium 8072 a paradoxical effect was observed. With this strain, the best synergic effect was detected at teicoplanin concentrations of 1-4 mg/L, but antibacterial activity was reduced at concentrations of 8, 16 and 32 mg/L. No synergic effect was observed with vancomycin. The combination of agar dilution with agar diffusion techniques may constitute a simple method for routine detection of synergic effects between glycopeptides and beta-lactams.

The addition of IFN-gamma to cultures of peripheral blood mononuclear cells (PBMCs) obtained from asymptomatic HIV-infected patients increased cell proliferation in response to HIV envelope synthetic peptides (Env), influenza A virus (VIRUS), and allogeneic lymphocytes (ALLO) but not to phytohaemagglutinin (PHA) stimulation. F(Ab)2 fragments of IgG purified from the sera of HIV-seropositive patients specifically interfered with IFN-gamma-induced cell proliferation in response to recall antigens. Neutralization of the lymphokine activity was found to be sustained by specific IFN-gamma antibodies. Data obtained demonstrate that IFN-gamma can restore the cell-mediated immunity of a number of asymptomatic HIV+ individuals in vitro, while IFN-gamma antibodies present in sera of patients with AIDS interfere with the activity of the lymphokine.

The principal design of a newly developed two-zone valved cracking phosphorus P2 molecular beam source with greatly improved performance based on InP thermal decomposition is outlined. Precise dimer phosphorus beam flux control is accomplished due to a thoughtfully designed and externally activated faucet placed between the InP decomposition zone and the cracking area of P4 vapors. Experimental tests show that the source can be easily incorporated into the standard ion-pumped molecular beam epitaxy (MBE) machine and can be used successfully for the MBE growth of device quality III–V single and multi-component phosphide epilayers incorporated into single- and multi-layer heterostructures with sharp interfaces.

Clostridium difficile-associated diarrhea is a significant problem in many hospitals and chronic care facilities (2). When established in the colon, pathogenic strains of C. difficile produce toxins that cause diarrhea and colitis; strains that do not produce toxins are not pathogenic. Toxin A (molecular mass, 308 kDa) acts as an enterotoxin, while toxin B (molecular mass, 250 to 270 kDa) acts as a cytotoxin (3), and both toxins may act synergistically in vivo. We report a case of isolation of a Clostridium sp. other than C. difficile that produced A and B toxins from an elderly patient belonging to a group of 17 subjects with diarrhea who were admitted to the same room of the orthopedics unit and all treated with ceftriaxone for 7 to 10 days of prophylaxis before operation. Diarrhea in this group of patients lasted for 20 days, but the patients were not simultaneously affected. Stool samples from all patients were collected in clear containers and screened for exotoxins A and B by a commercially available enzyme immunoassay (Ridascreen C. difficile toxin A, B; R-Biopharm, Darmstadt, Germany). Swabs of stool specimen from each patient were transported in an anaerobic system (Port-A-Cul; BBL) and inoculated onto C. difficile selective agar plates (Oxoid) prepared in-house. The plates were then incubated anaerobically for 48 to 72 h at 35°C. Colonies were identified as Clostridium by colonial morphology, Gram’s stain, and biochemical profile (API 20 A anaerobe and Vitek ANI systems; bio-Merieux). Out of samples from 17 patients, toxins A and B were detected at different times in the stools of only 8 patients, but no C. difficile was isolated. In our experience, this is very unusual, because we generally isolate C. difficile from stools with toxins A and B. For the last 3 of the 17 patients we therefore considered all the clostridia isolated, namely, C. paraputrificum, C. baratii, C. tertium, C. bifermentans, C. innocuum, C. clostridiiforme, and C. sporogenes. After identification the clostridium strains were cultured and stored, before being checked for the presence of toxigenic products, in Bactec plus anaerobic vials (Becton Dickinson) containing liquid medium and matrix with fluorescent material that reacts to CO2 produced by bacteria. This reaction, as measured by the photodetectors of an instrument, is evidence of bacterial growth. We chose this method because the bacterial culture is performed outside an anaerobic chamber and we observed (unpublished observation) that the production of bacterial exotoxins may be predicted by the black coloring of matrix at the bottom of the vials. Indeed, the bacterial cultures of nontoxigenic strains produced no change in color and the matrix remained brown, while exotoxin-producing strains changed the matrix to black. Only in one of the last three patients considered did we identify a strain of C. baratii producing exotoxins A and B. Cultures of this microorganism were positive for A and B exotoxin production by enzyme immunoassay and for cytotoxin B by tissue culture assay using Vero cells and C. difficile antitoxin B (Biolife) as a control. After the addition of a 0.5-ml inoculum (0.5 McFarland) in 30 ml of liquid medium, C. difficile ATCC 9689 produced black coloring of the matrix in 3 days while C. baratii produced black coloring in 8 days. This Clostridium sp. was positive for C. difficile antigens by a latex agglutination test (Meridian Diagnostics Inc.), but its biochemical profile and gas chromatography results were different from those of C. difficile ATCC 9689 and similar to those of a C. baratii toxin nonproducer. For toxigenic C. baratii tested by an agar dilution method (4), the MICs of teicoplanin and metronidazole were 1 and 8 mg/liter, respectively. The patients with diarrhea were treated at first with metronidazole without success and later with teicoplanin. This experience suggests that Clostridium microorganisms other than C. difficile sometimes produce A and B toxins but also that C. baratii, which is reported as a botulinal toxin producer (1, 5), is capable of toxin A and B production.

The increased social and economic importance of ready–made food production, together with the complexity of production technology, processing, handling and acceptance of these fragile and perishable products requires extensive knowledge of their physical properties. Viscoelastic properties play an important role in the handling and quality attributes of creams.Our study was to investigate the rheological properties of different confectionary creams, by scanning the field of shear rates at constant temperature and frequency, angular frequency scanning at small deformations and quantification of rheological changes during application of deformation voltages. The creams tested were made in the laboratory using specific concentrates as fine powders, marketed by the company “Dr. Oetker” compared with similar creams based on traditional recipes and techniques. Following the researches conducted we could conclude that both traditional creams and the instant ones are semi fluid food products with pseudoplastic and thixotropic shear flow behavior, with structural viscosity. Instant and traditional creams behaved as physical gels with links susceptible to destruction, when subjected to deformation forces.

Since the 1990s, when the news about antioxidants and their benefits to health has begun to spread to the general public, statements about the benefits of antioxidants ranged from preventing colds to cancer treatment. Fruits and vegetables are excellent sources of antioxidants. Since the beginning of the 21st century, goji berries have become increasingly popular in Europe and North America and have been promoted in advertisements and in the media as an anti-aging remedy. Goji is a relatively new name given to Lycium Barbarum and Lycium chinense, two nearby species, with a long history of use as medicinal and food plants in East Asia, particularly in China. In the present paper are presented analysis results of Goji fruits and food products made from goji fruits (jam and jelly). Storage conditions are important factors for jams and jelly quality. The objective of this study was to monitor the physicochemical stability, antioxidant activity and sensorial profile of goji fruits jam and jelly. Special attention was paid to total phenolic and flavonoid content, antioxidant activity, total soluble solids, titratable acidity, pH and sensorial characteristics. Our results showed the antioxidant activity of the goji fruit, values which correlate well with the results obtained for total phenolic (351±7.25 mg GAE/100g ) and flavonoid content (53.06±1.23 mg QE/100g). The antioxidant activity of the goji fruits was maintained also in the finished products obtained in the present study jam (60.98 %) and jelly (41.96 %). Both goji fruits jam and goji fruits jelly showed no significant variations of physico-chemical characteristics and sensorial parameter scores after storage at refrigeration temperature for 10 days.