Metabolomic based approach to identify biomarkers of broccoli intake.
It is well-established that consumption of cruciferous and brassica vegetables has a correlation with reduced rates of many negative health outcomes. There is an increased interest in identifying food intake biomarkers to address limitations related to self-reported dietary assessment. The study aims to identify biomarkers of broccoli intake using metabolomic approaches, examine the dose-response relationship, and predict the intake by multimarker panel. Eighteen volunteers consumed cooked broccoli in A-Diet Discovery study and fasting and postprandial urine samples were collected at 2, 4 and 24 hours. Subsequently the A-Diet Dose-response study was performed where volunteers consumed different portions of broccoli (49, 101 or 153 g) and urine samples were collected at the end of each intervention week. Urine samples were analysed by 1H-NMR and LC-MS. Multivariate data analysis and one-way ANOVA were performed to identify discriminating biomarkers. A panel of putative biomarkers was examined for its ability to predict intake through a multiMarker model. Multivariate analysis revealed discriminatory spectral regions between fasting and fed metabolic profiles. Subsequent time-series plots revealed multiple features increased in concentration following the consumption. Urinary S-methyl cysteine sulfoxide (SMCSO) increased as broccoli intake increased (0.17-0.24 μM per mOSM per kg, p < 0.001). Similarly from LC-MS data genipin, dihydro-β-tubaic acid and sinapic acid increased with increasing portions of intake. A panel of 8 features displayed good ability to predict intake from biomarker data only. In conclusion, urinary SMCSO and several LC-MS features appeared as potentially promising biomarkers of broccoli consumption and demonstrated dose-response relationship. Future work should focus on validating these compounds as food intake biomarkers.


[Estimation of ethanol concentrations in biological media. Comparison of alkylnitrite and direct vapor-phase methods].
Ethanol assays in the blood, saliva and urine were made according to two methods: alkylnitrite gas-liquid chromatography and direct vapor-phase method without thermostating. N-propanol was used as an internal standard. For saliva and urine the methods produced similar results. For blood, the direct vapor-phase method detected significantly lower level of ethanol compared to alkylnitrite one (p = 0.001). 24-h storage of the samples in refrigerator does not deviate the results. Recommendations are provided for most effective usage of each method.


Inhibition of Jurkat T Cell Growth by N-farnesyl-norcantharimide Through Up-regulation of Tumor Suppressor Genes and Down-regulation of Genes for Steroid Biosynthesis, Metabolic Pathways and Fatty Acid Metabolism.
To evaluate the anti-cancer mechanism of N-Farnesyl-norcantharimide (NC15). The viability of NC15-treated human leukemic Jurkat T (JKT) cells was assessed using the Kit-8 cell counting method. Flow cytometry analysis, human apoptosis antibody array assay, and whole genome sequencing were adopted to investigate the mechanism underlying the anti-cancer activity of NC15 in JKT cells. The growth inhibition rates of NC15 in JKT cells were about 80% and 95% after treatment with 8 μmol/l NC15 for 24 and 48 h, respectively. The percentages of NC15-treated JKT cells in the sub-G1 phase at 24 and 48 h were 22.0% and 34.3%, respectively, in contrast to the 1.5% in the control. Next-generation sequencing showed that many tumor suppressor genes (TSG) were up-regulated, while many genes associated with steroid biosynthesis, metabolic pathways, and fatty acid metabolism were down-regulated. NC15 can reduce the cell viability and increase the percentage of JKT cells in the sub-G1 phase by up-regulating TSG and related genes, and down-regulating the genes for steroid biosynthesis, metabolic pathways and fatty acid metabolism, instead of through apoptosis.


Prognostic factors in patients with advanced gastric cancer with macroscopic invasion to adjacent organs treated with radical surgery.
BACKGROUND: The prognosis of patients with gastric cancer with invasion to adjacent organs is poor. The prognostic factors of patients with advanced gastric cancer with macroscopic invasion to adjacent organs (T4) who were treated with radical surgery was determined in the present study.METHODS: A total of 86 consecutive patients with advanced gastric cancer who underwent radical (potentially curable) gastrectomy with combined resection of other organs for macroscopic invasion to adjacent organs during surgery, were investigated. The organs invaded macroscopically were the pancreas in 43 patients, mesocolon in 29, liver in 7, transverse colon in 5, adrenal gland in 3, spleen in 1, diaphragm in 1, and other organs in 5. The prognostic factors were evaluated by univariate and multivariate analysis.RESULTS: The cumulative 5-year survival rate of the patients treated by radical surgery with the combined resection of invaded organs was 35.0%. Multivariate analysis demonstrated that location of the tumor, lymph node metastasis, histological depth of invasion, and extent of lymph node dissection were significant prognostic factors in advanced gastric cancer patients treated by radical surgery with combined resection of adjacent organs for macroscopic invasion.CONCLUSION: For patients with macroscopic T4 gastric cancer located in the middle- or lower-third of the stomach, aggressive resection of invaded adjacent organs with extended lymph node dissection should be performed to improve long-term outcome.


Phosphate Limitation Triggers the Dissolution of Precipitated Iron by the Marine Bacterium Pseudovibrio sp. FO-BEG1.
Phosphorus is an essential nutrient for all living organisms. In bacteria, the preferential phosphorus source is phosphate, which is often a limiting macronutrient in many areas of the ocean. The geochemical cycle of phosphorus is strongly interconnected with the cycles of other elements and especially iron, because phosphate tends to adsorb onto iron minerals, such as iron oxide formed in oxic marine environments. Although the response to either iron or phosphate limitation has been investigated in several bacterial species, the metabolic interplay between these two nutrients has rarely been considered. In this study we evaluated the impact of phosphate limitation on the iron metabolism of the marine bacterium Pseudovibrio sp. FO-BEG1. We observed that phosphate limitation led to an initial decrease of soluble iron in the culture up to three times higher than under phosphate surplus conditions. Similarly, a decrease in soluble cobalt was more pronounced under phosphate limitation. These data point toward physiological changes induced by phosphate limitation that affect either the cellular surface and therefore the metal adsorption onto it or the cellular metal uptake. We discovered that under phosphate limitation strain FO-BEG1, as well as selected strains of the Roseobacter clade, secreted iron-chelating molecules. This leads to the hypothesis that these bacteria might release such molecules to dissolve iron minerals, such as iron-oxyhydroxide, in order to access the adsorbed phosphate. As the adsorption of phosphate onto iron minerals can significantly decrease phosphate concentrations in the environment, the observed release of iron-chelators might represent an as yet unrecognized link between the biogeochemical cycle of phosphorus and iron, and it suggests another biological function of iron-chelating molecules in addition to metal-scavenging.


Is combined topical and intravenous tranexamic acid superior to intravenous tranexamic acid alone for controlling blood loss after total hip arthroplasty?: A meta-analysis.
We performed a meta-analysis of randomized controlled trials (RCTs) to compare the efficacy and safety of combined intravenous (IV) and topical tranexamic acid (TXA) with IV-TXA alone for controlling blood loss in patients following primary total hip arthroplasty (THA). PubMed, EMBASE, the Cochrane Central Register of Controlled Trials, the Google database, the Chinese Wanfang database, and the China National Knowledge Infrastructure database were searched to identify studies comparing combined IV and topical TXA with IV-TXA alone in patients who were prepared for THA. The weighed mean differences for total blood loss, hemoglobin drop, intraoperative blood loss, and the length of hospital stay were calculated. We calculated risk ratios for the need for transfusion and the occurrence of deep venous thrombosis (DVT) in the combined TXA and IV-TXA alone groups. Relevant data were analyzed using Reviewer Manager 5.3.0. Eight RCTs with a total of 850 patients (combined TXA: n = 471; IV-TXA: n = 479) were included in this meta-analysis. Pooled results indicated that compared with the IV-TXA alone group, the combined TXA group was associated with a lesser need for transfusion, total blood loss, intraoperative blood loss, and hemoglobin drop (P < .05). There was no significant difference between the 2 groups for the length of hospital stay and the occurrence of DVT (P > .05). The current meta-analysis indicated that combined topical and IV-TXA was a relatively effective hemostasis method compared with IV-TXA alone. The number of studies included in this meta-analysis is limited, and more studies are needed to verify the effects of combined IV and topical TXA in THA patients.


Importance of genetic testing in unexplained cardiac arrest.
Genetic testing is recommended in specific inherited heart diseases but its role remains unclear and it is not currently recommended in unexplained cardiac arrest (UCA). We sought to assess the yield and clinical utility of genetic testing in UCA using whole-exome sequencing (WES). Survivors of UCA requiring external defibrillation were included from the Cardiac Arrest Survivor with Preserved Ejection fraction Registry. Whole-exome sequencing was performed, followed by assessment of rare variants in previously reported cardiovascular disease genes. A total of 228 UCA survivors (mean age at arrest 39 ± 13 years) were included. The majority were males (66%) and of European ancestry (81%). Following advanced clinical testing at baseline, the likely aetiology of cardiac arrest was determined in 21/228 (9%) cases. Whole-exome sequencing identified a pathogenic or likely pathogenic (P/LP) variant in 23/228 (10%) of UCA survivors overall, increasing the proportion of 'explained' cases from 9% only following phenotyping to 18% when combining phenotyping with WES. Notably, 13 (57%) of the 23 P/LP variants identified were located in genes associated with cardiomyopathy, in the absence of a diagnosis of cardiomyopathy at the time of arrest. Genetic testing identifies a disease-causing variant in 10% of apparent UCA survivors. The majority of disease-causing variants was located in cardiomyopathy-associated genes, highlighting the arrhythmogenic potential of such variants in the absence of an overt cardiomyopathy diagnosis. The present study supports the use of genetic testing including assessment of arrhythmia and cardiomyopathy genes in survivors of UCA.


Optimal Tuning Perspective of Range-Separated Double Hybrid Functionals.
We study the optimal tuning of the free parameters in range-separated double hybrid functionals, based on enforcing the exact conditions of piecewise linearity and spin constancy. We find that introducing the range separation in both the exchange and the correlation terms allows for the minimization of both fractional charge and fractional spin errors for singlet atoms. The optimal set of parameters is system specific, underlining the importance of the tuning procedure. We test the performance of the resulting optimally tuned functionals for the dissociation curves of diatomic molecules. We find that they recover the correct dissociation curve for the one-electron system, H2+, and improve the dissociation curves of many-electron molecules such as H2 and Li2, but they also yield a nonphysical maximum and only converge to the correct dissociation limit at very large distances.


From mindful attention to social connection: The key role of emotion regulation.
Effective emotion regulation is important for high-quality social functioning. Recent laboratory-based evidence suggests that mindfulness may enhance emotion regulation in socioemotional contexts; however, little is known about mindful emotion regulation during in vivo social interactions. In a study of romantic couples, we assessed each partner's mindfulness and top-down attentional efficiency (with an Emotional Go/No-Go task) prior to sampling emotions and perceived connection with others during day-to-day social interactions. Analyses revealed that mindfulness-related differences in top-down attentional efficiency on the Emotional Go/No-Go predicted positive emotion during daily social interactions. In turn, positive emotion and two additional indices of social emotion regulation each mediated the relation between actor mindfulness and perceived social connection. In corresponding analyses, neither trait reappraisal nor suppression use predicted the outcomes, and all mindfulness relations held controlling for these strategies. Findings support a framework for investigating mindfulness and higher-quality social functioning, for which mindful emotion regulation may be key.


Functional characterization of the polyketide synthase gene required for ochratoxin A biosynthesis in Penicillium verrucosum.
The ochratoxin A (OTA) polyketide synthase otapks gene has been cloned from Penicillium verrucosum. A P. verrucosum mutant in which the otapksPV gene has been interrupted cannot synthesize ochratoxin A. The protein is most similar to the citrinin polyketide synthase CtnpksMa from Monascus anka (83% identity at the amino acid level). Different nutritional conditions influence OTA production in P. verrucosum, with the addition of glycerol and galactose to MCB resulting in approximately 19 and 32 fold increases in OTA production respectively. These effects are mirrored in increased levels of otapksPV gene transcription. In contrast, the addition of glucose to MCB containing galactose results in an approximate 10 fold repression in OTA production, with this repression again being mirrored in decreased levels of otapksPV gene transcription. Thus the effects of different carbon sources on OTA production in P. verucosum appear to be regulated at the level of gene transcription. Two additional open reading frames, otaE and otaT, were identified in the 5' and 3' flanking regions of otapksPV, respectively. The otaT and otaE genes are co-expressed with P. verrucosum otapksPv, indicating a possible role for these genes in OTA biosynthesis. Furthermore, otaT and otaE were identified as putative homologues of the M. anka citrinin transporter ctnC (72% amino acid identity) and M. anka citrinin oxidoreductase ctnB (83% amino acid identity); suggesting that the genes involved in OTA production in P. verrucosum may be very similar to those involved in citrinin production in M. anka.


Tailoring Intercalant Assemblies at the Graphene-Metal Interface.
The influence of graphene on the assembly of intercalated material is studied using low-temperature scanning tunneling microscopy. Intercalation of Pt under monolayer graphene on Pt(111) induces a substrate reconstruction that is qualitatively different from the lattice rearrangement induced by metal deposition on Pt(111) and, specifically, the homoepitaxy of Pt. Alkali metals Cs and Li are used as intercalants for monolayer and bilayer graphene on Ru(0001). Atomically resolved topographic data reveal that at elevated alkali metal coverage (2 × 2)Cs and (1 × 1)Li intercalant structures form with respect to the graphene lattice.


Structure of aspartate β-semialdehyde dehydrogenase from Francisella tularensis.
Aspartate β-semialdehyde dehydrogenase (ASADH) is an enzyme involved in the diaminopimelate pathway of lysine biosynthesis. It is essential for the viability of many pathogenic bacteria and therefore has been the subject of considerable research for the generation of novel antibiotic compounds. This manuscript describes the first structure of ASADH from Francisella tularensis, the causative agent of tularemia and a potential bioterrorism agent. The structure was determined at 2.45 Å resolution and has a similar biological assembly to other bacterial homologs. ASADH is known to be dimeric in bacteria and have extensive interchain contacts, which are thought to create a half-sites reactivity enzyme. ASADH from higher organisms shows a tetrameric oligomerization, which also has implications for both reactivity and regulation. This work analyzes the apo form of F. tularensis ASADH, as well as the binding of the enzyme to its cofactor NADP+.


Initial-value-problem solution for isolated rippled shock fronts in arbitrary fluid media.
Following the work of Roberts [Los Alamos Scientific Laboratory Report No. LA-299, 1945 (unpublished)], we investigate the effect of small two-dimensional perturbations on an isolated, planar shock front moving steadily through an inviscid fluid medium with an arbitrary equation of state (EOS). In the context of an initial-value problem, we derive explicit analytical expressions for the linearized, time-dependent Fourier coefficients associated with an initial corrugation of the front. The temporal evolution of these coefficients superficially resembles the attenuated "ringing" of a damped harmonic oscillator, but with the important distinctions that the frequency of oscillation is not constant, and that the damping factor is not simply an exponential function of time t. It is shown that at least two three-parameter families of stable solutions exist, one more strongly damped than the other. In both cases, we find that the envelope of oscillations decays asymptotically as t(-3/2), with shorter wavelengths dying out earlier than longer ones. For a particular perturbed-shock system, the strength of the front and the EOS properties of the material through which it propagates determine the applicable family of solutions. Theoretical predictions agree well with FAST2D numerical simulations for several examples derived from the CALEOS library.


Careers beyond the perioperative environment.
Perioperative nursing has adapted to meet the needs of new health care delivery models. Opportunities are present for perioperative nurses not only as caregivers, but also as professionals with the ability to transition into nontraditional positions. A prospective employee needs to consider many factors before making a career-changing decision.


An unusual case of globe-sparing penetrating orbital injury by a nail.
We describe a case of a 28-month-old boy who presented to our emergency department with a right orbital penetration by an iron nail. At presentation, no pain or functional loss were noted. Plain head radiographs and computed tomography were performed and demonstrated a two centimeters long nail penetrating the right orbit. The foreign body was extracted carefully in the operating room. No complications were observed upon follow-up. Rarely, orbital penetration by an iron-nail can present without functional disturbances or pain and is not necessarily accompanied by sequelae.


Postoperative chemo-radiotherapy with temodal in patients with glioblastoma multiforme--survival rates and prognostic factors.
Glioblastoma multiforme (GM) is the most malignant histological type of brain tumors. It affects the active age and independent of the applied general therapeutic methods in oncology the problem with the short survival of the patients still remains. In the recent years simultaneous application of Temodal and radiation therapy was introduced in medical practice. AIM OF THE PRESENT STUDY: To compare the survival rate of a group of patients subjected to radiation therapy only to the survival rate of a group subjected to simultaneous treatment with Temodal and radiation therapy (RT). 179 patients with GM were divided into 2 groups and subjected to radiation therapy with realized total dose of 60 Gy fractionated into 5 x 2 Gy weekly. 44 of the patients underwent chemo-radiotherapy with Temodal dosed 75 mg/m2, and the rest 135 ones received RT alone. All patients received corticosteroid treatment. The Karnofski status (KPS) of the group with Temodal treatment was 90-100%. Survival rates were assessed using the Kaplan-Mayer method and side effects for-the group with simultaneous chemo-radiotherapy. Chemo-radiation therapy was well tolerated by all patients. Median survival time was 14.83 months for the group with Temodal and 14.67 months for the other group. The progression-free time was 8.5 months. Vomiting was observed in 11.4% of the patients and was corrected with antiemetics. No side hematological deviations were developed as well as pneumonias. Temozolomide in combination with chemotherapy is a gold standard for GM patients and it shows a tendency of increased survival. The clinical tolerance is good and no marked side reactions are observed


Monitoring minimal residual disease using chromosomal translocations in childhood ALL.
Clonal chromosomal abnormalities have been identified in approximately 80% of childhood ALL. In most instances the genes disrupted by these abnormalities have been identified, thus providing important insights into disease pathogenesis and normal cellular physiology. Polymerase chain reaction (PCR) amplification of fusion transcripts resulting from chromosomal translocations has emerged as a sensitive and reproducible method to monitor minimal residual disease (MRD) in childhood ALL. The measure of the initial response to therapy in patients who have achieved complete remission by morphological standards can dissect clinical heterogeneity within the genetically homogeneous childhood ALL subgroup. Moreover, MRD monitoring can be applied to predict impending relapses early. Despite notable progress with this method, several critical issues must be resolved before MRD determinations can be routinely considered in clinical decision making. This chapter will focus on the main progress and common pitfalls in the PCR detection of chromosomal translocations applied to clinical studies.


In-situ AFM studies of the phase-transition behavior of single thermoresponsive hydrogel particles.
The volume phase transition (VPT) behavior of individual thermally responsive poly(N-isopropylacrylamide-co-acrylic acid) (pNIPAm-co-AAc) hydrogel microparticles was studied by in-situ dynamic mode atomic force microscopy (AFM) and force spectroscopy during heating and cooling cycles. Hydrogel samples were prepared by electrostatic immobilization of microparticles to amine-modified gold surfaces. The AFM studies of particle deswelling were performed by varying the force applied on the particles during imaging as a function of the geometry and material of the AFM probe. Aluminum-coated silicon cantilevers were found to influence substantially the behavior of the particles during the VPT, leading to a significant shape change. Low force impact magnetic excitation of the AFM probe (MAC mode) during dynamic mode measurements resulted in an undisturbed deswelling behavior enabling observation of the expected volume changes of the particles without significant tip-sample interaction. Hence, MAC-mode AFM was determined to be the most suitable technique for in-situ AFM studies on volume and shape changes at single hydrogel particles during VPT. Elasticity measurements performed at single particles at temperatures below and above the VPT revealed a 15-fold increase in the Young's modulus after passing the VPT, indicating the transition from a soft, swollen network to a stiffer, deswollen state.


Well-dispersed Au co-catalyst deposited on a rutile TiO2 photocatalyst via electron traps.
We deposited Au nanoparticles as a co-catalyst onto a TiO2 photocatalyst by reducing [AuCl4]- using electrons trapped in the oxygen vacancies of TiO2. The dispersibility and hydrogen production ability of the Au co-catalyst are higher than those prepared using the conventional photodeposition method.


Organotin and organogermanium linkers for simple, direct functionalization of polyoxotungstates.
Fifteen Keggin-anion-derived polytungstates [TW11O39[MCH2CH2X]]n- (T = Si, Ge, Ga; M = Sn, Ge; X = COOH, COOCH3, CONH2, CN; n = 5, 6) were prepared in aqueous or aqueous-organic solution from the corresponding lacunary polytungstates and trichlorotin and -germanium precursors, and were isolated as caesium salts. The derivatized polytungstates were characterized by elemental analysis, multinuclear NMR spectroscopy, and cyclic voltammetry; they are stable in aqueous solution to pH 6-7. NMR spectroscopy revealed the presence of a second (beta1 or beta3) isomer in the tungstogallate derivatives. Acid hydrolysis of the ester and nitrile derivatives could be achieved without decomposition of the polytungstate moieties, and esterification and amidation of the carboxylate functions was straightforward using standard coupling techniques, e.g. the formation, isolation and characterization of [SiW11O39[Ge(CH2)2CONHCH2COOCH3]]5- from glycine methyl ester. Since the Cl3MCH2CH2X precursors are readily accessible by hydrostannation/germanation reactions with the corresponding alkenes, novel coupled polytungstates, such as [(SiW11O39GeCH2CH2COOCH2)4C]20- from pentaerythritol tetraacrylate, can also be prepared.


Coping with fraud.
The tardiness of actions by Harvard officials when faced with confessed fraudulent research conducted by John Darsee is contrasted with the more rapid responses by authorities in the cases of William T. Summerlin (Sloan-Kettering) and Vijay Soman (Yale).


Robust Chalcogenophene Viologens as Anolytes for Long-Life Aqueous Organic Redox Flow Batteries with High Battery Voltage.
A series of chalcogenophene viologens ([(NPr)2FV]Cl4, [(NPr)2TV]Cl4, and [(NPr)2SeV]Cl4) as anolytes for neutral aqueous organic redox flow batteries (AORFBs) via a combination of chalcogenophenes (furan, thiophene, and selenophene) and viologens are reported. The chalcogenophene viologens showed narrow HOMO-LUMO energy gap, high solubility, and stable electrochemical properties. Compared with the parent [(NPr)2V]Cl4, the introduction of π-conjugated chalcogenophene groups reduced the redox potential and enhanced the stability of their free radical state, which endowed the chalcogenophene viologens/FcNCl-based AORFBs with a higher theoretical battery voltage of 1.20 V and enhanced stability for one-electron storage. In particular, the [(NPr)2FV]Cl4/FcNCl-based AORFB exhibited excellent long-cycle stability for 3000 cycles with 0.0006% capacity decay per cycle for one-electron storage and 300 cycles with 0.06% capacity decay per cycle for two-electron storage at a charge voltage of 1.9 V (1.42 V theoretical battery voltage). This work provided a new strategy for regulating the voltage and improving the performance of neutral AORFBs.


Protective influences of families for children adopted from the former Soviet Union.
To (a) characterize the total competence of 9- to 12-year old children adopted from the former Soviet Union who have resided in the United States at least 5 years, and (b) evaluate risks and protective influences of adoptive families and their relationships to competence. Longitudinal, descriptive study. In the previously reported phase, the families of 105 internationally adopted children and their families from 23 U.S. states were assessed via telephone interviews and postal survey. In this phase, 3.5 years later, 46 of the families from 16 states were located and reassessed. Measures included: Total Competence scale of the Child Behavior Checklist (CBCL), Family Environment Scale (FES), and demographic information. Multivariate regression analysis was conducted to determine which risk and protective factors contributed to competence scores of the adopted children. Between Time 1 (mean age of children, 7.7 years) and Time 2 (mean age of children, 11 years), average scores on the Total Competence scale of the CBCL did not change significantly, despite impending challenges of early adolescence. Subdomain scores on the FES were more positive than norms. Results from the regression analysis showed that, of the risk and protective factors considered, only birth weight and the cohesion subscale of the FES were statistically significant in explaining variation in the competence score of the CBCL. Families continued to face challenges, but findings were consistent with other studies showing that, despite early adversities, these adopted children generally fared well developmentally with protective family environments of major importance.


Investigating homology between proteins using energetic profiles.
Accumulated experimental observations demonstrate that protein stability is often preserved upon conservative point mutation. In contrast, less is known about the effects of large sequence or structure changes on the stability of a particular fold. Almost completely unknown is the degree to which stability of different regions of a protein is generally preserved throughout evolution. In this work, these questions are addressed through thermodynamic analysis of a large representative sample of protein fold space based on remote, yet accepted, homology. More than 3,000 proteins were computationally analyzed using the structural-thermodynamic algorithm COREX/BEST. Estimated position-specific stability (i.e., local Gibbs free energy of folding) and its component enthalpy and entropy were quantitatively compared between all proteins in the sample according to all-vs.-all pairwise structural alignment. It was discovered that the local stabilities of homologous pairs were significantly more correlated than those of non-homologous pairs, indicating that local stability was indeed generally conserved throughout evolution. However, the position-specific enthalpy and entropy underlying stability were less correlated, suggesting that the overall regional stability of a protein was more important than the thermodynamic mechanism utilized to achieve that stability. Finally, two different types of statistically exceptional evolutionary structure-thermodynamic relationships were noted. First, many homologous proteins contained regions of similar thermodynamics despite localized structure change, suggesting a thermodynamic mechanism enabling evolutionary fold change. Second, some homologous proteins with extremely similar structures nonetheless exhibited different local stabilities, a phenomenon previously observed experimentally in this laboratory. These two observations, in conjunction with the principal conclusion that homologous proteins generally conserved local stability, may provide guidance for a future thermodynamically informed classification of protein homology.


Chronic autoimmune disorders are increased in coeliac disease: A case-control study.
Coeliac disease (CD) is an autoimmune disorder of the small bowel associated with increased risk of additional autoimmune diseases (ADs).To investigate the prevalence of ADs in a population of adult coeliac patients.This was a retrospective case-control study. Data from coeliac patients and controls referred to a tertiary center between 2013 and 2016 were collected. The frequency of ADs and the unadjusted and adjusted odds ratios (ORs) for age, gender, disease duration, and body mass index with their 95% confidence intervals (CIs) were evaluated.Two hundred fifty-five patients with CD (median age 37.1 years; 206 women) were matched with 250 controls. ADs were more frequent (35.3%) in coeliac patients than in controls (15.2%). Adjusted ORs for the presence of only 1, at least 1, and more than 1 AD were 3.13 (95% CI 1.81-5.42, P < .0001), 3.31 (95% CI 2.00-5.46, P < .0001), and 3.93 (95% CI 1.49-10.36, P = .006), respectively. Hashimoto thyroiditis was the most prevalent AD (24.3% vs. 10%) OR = 2.55 (95% CI 1.39-4.70, P < .0001), followed by psoriasis (4.3% vs. 1.6%), type 1 diabetes (2.7% vs. 0.4%), and Sjögren syndrome (2.4% vs. 0.4%).These findings suggest a need for a careful surveillance of autoimmune status, especially for Hashimoto thyroiditis in patients with celiac disease.


Activity and stability of recombinant human superoxide dismutase in buffer solutions and hypothermic perfusates.
The stability of recombinant human superoxide dismutase (r-hSOD) in buffer solutions was studied in solutions at various pH and temperatures. Additionally, we studied the effects of incubation with proteases, serum and two types of hypothermic perfusates. R-hSOD was stable in the pH range of 6-11 and at temperatures up to 80 degrees C for 30 min. R-hSOD activity was not affected by incubation with trypsin, aminopeptidase M or serum for 2 h. R-hSOD activity determined at various temperatures (4-37 degrees C) did not vary remarkably. R-hSOD in hypothermic perfusates was stable at 4-37 degrees C for 24 h.


Ultrasound evaluation of fetal nasal bone at 11 to 14 weeks in a consecutive series of 1906 fetuses.
The aim of this study is to evaluate the significance of nasal bone ossification as a marker for trisomy 21 at 11 to 14 weeks' gestation in an unselected obstetric population referred to our Centre. A total of 1906 consecutive fetuses undergoing nuchal translucency scan at 11 to 14 weeks' gestation were evaluated for the presence of hypoplasia/absence of nasal bone. The data obtained were correlated with fetal karyotype. A successful view of the fetal profile was obtained in 1752 fetuses (91.9%). The nasal bone was hypoplastic/absent in 12 of 19 fetuses with chromosomal abnormalities. There were 10 cases of trisomy 21, in 8 of which hypoplastic/absent nasal bone was observed. Furthermore, absence of nasal bone was recorded in 24 of 1733 chromosomally normal fetuses. Nasal bone evaluation may improve the detection of trisomy 21 in the first trimester in an unselected obstetric population. Although numerically limited, our experience confirms that delayed nasal bone ossification (hypoplasia/absence of nasal bone) is rarely observed in chromosomally normal fetuses (1.4%). An appropriate training of operators is mandatory in order to achieve an acceptable performance.


Effects of (+)-cyanidanol-3 on chronic liver injury.
Thirty patients of both sexes, suffering from alcoholic or diabetic liver injury, were studied in order to establish therapeutic effects of daily 3000 mg (+)-cyanidanol-3 administered for 3 weeks. In addition to subjective complaints, alterations in liver-specific enzyme-levels, changes in serumlipoids, the BSP half-time, activity of the microsomal enzyme-system and changes in the diabetic metabolism were studied. During the observation significant improvement of the disturbed enzyme-activities, decrease of the high lipoid level, reduction of the BSP half-time and the increasing activity of the microsomal enzyme-system could be proved. In case of diabetic patients a favourable change of the metabolic status could be established. Subjective complaints of patients subsided. In case of 17 patients the radionuclide HIDA method was applied too in order to study the dynamics of bile-transport. A moderately significant acceleration of the bile-transport could be observed. While continuing the treatment of 15 patients for 6 months, a control liver biopsy was carried out and in 10 cases it revealed favourable changes in the histological finding. During cyanidanol treatment gastrointestinal intolerance was observed in two patients, but only temporarily. Immune haemolysis did not occur.


Toxic effects of cannabis and cannabinoids: Animal data.
The present article reviews the main toxic effects of cannabis and cannabinoids in animals. Toxic effects can be separated into acute and chronic classifications. Acute toxicity studies show that it is virtually impossible to die from acute administration of marijuana or tetrahydrocannabinol, the main psychoactive component of cannabis. Chronic toxicity involves lesions of airway and lung tissues, as well as problems of neurotoxicity, tolerance and dependence, and dysregulations in the immune and hormonal systems. Animal toxicity data, however, are difficult to extrapolate to humans.


Co-delivery nanoparticles with characteristics of intracellular precision release drugs for overcoming multidrug resistance.
Combination chemotherapy in clinical practice has been generally accepted as a feasible strategy for overcoming multidrug resistance (MDR). Here, we designed and successfully prepared a co-delivery system named S-D1@L-D2 NPs, where denoted some smaller nanoparticles (NPs) carrying a drug doxorubicin (DOX) were loaded into a larger NP containing another drug (vincristine [VCR]) via water-in-oil-in-water double-emulsion solvent diffusion-evaporation method. Chitosan-alginate nanoparticles carrying DOX (CS-ALG-DOX NPs) with a smaller diameter of about 20 nm formed S-D1 NPs; vitamin E D-α-tocopheryl polyethylene glycol 1000 succinate-modified poly(lactic-co-glycolic acid) nanoparticles carrying VCR (TPGS-PLGA-VCR NPs) with a larger diameter of about 200 nm constituted L-D2 NPs. Some CS-ALG-DOX NPs loaded into TPGS-PLGA-VCR NPs formed CS-ALG-DOX@TPGS-PLGA-VCR NPs. Under the acidic environment of cytosol and endosome or lysosome in MDR cell, CS-ALG-DOX@TPGS-PLGA-VCR NPs released VCR and CS-ALG-DOX NPs. VCR could arrest cell cycles at metaphase by inhibiting microtubule polymerization in the cytoplasm. After CS-ALG-DOX NPs escaped from endosome, they entered the nucleus through the nuclear pore and released DOX in the intra-nuclear alkaline environment, which interacted with DNA to stop the replication of MDR cells. These results indicated that S-D1@L-D2 NPs was a co-delivery system of intracellular precision release loaded drugs with pH-sensitive characteristics. S-D1@L-D2 NPs could obviously enhance the in vitro cytotoxicity and the in vivo anticancer efficiency of co-delivery drugs, while reducing their adverse effects. Overall, S-D1@L-D2 NPs can be considered an innovative platform for the co-delivery drugs of clinical combination chemotherapy for the treatment of MDR tumor.


Relationships between Physical Activity and Selected Chronic Diseases among Functionally Independent Long-Term Care Residents during the Post-Lockdown Period in Croatia.
The aim of this observational study was to investigate the level and association of physical activity and selected chronic diseases in functionally independent LTC residents after prolonged physical and social isolation during COVID-19 in Croatia. Adhering to the inclusion criteria, 180 functionally independent residents were included in the study. Assessment of physical activity was carried out by 7-day motor monitoring. Prolonged physical and social isolation negatively affected the achieved level of physical activity of LTC residents (x¯ = 5058.74). Major depressive disorder resulted in significantly lower residents' physical activity scores, demonstrating a shrinking effect ranging from 0.42 to 0.45. A significant negative impact on the residents' physical activity was also found in the presence of osteoarthritis and iron deficiency anemia, where a downward effect was present in the range from 0.66-0.72 and 0.64 to 0.66. The presence of comorbidities has a significant negative impact on the residents' physical activity, where a downward effect is present in the range from 0.91-0.92.


Banded structure in solitary trichoepithelioma.
Electron microscopical observations were made in a case of solitary trichoepithelioma. Enlarged ERs were found in the tumor cells, showing a peculiar structure with fine granules and an electron-dense banded structure, about 50 nm wide respectively, lying at intervals of 250 nm. In addition to this structure, dense ovoid bodies, about 500 to 600 nm in diameter, were also observed.


Temperature dependence of protein folding kinetics in living cells.
We measure the stability and folding rate of a mutant of the enzyme phosphoglycerate kinase (PGK) inside bone tissue cells as a function of temperature from 38 to 48 °C. To facilitate measurement in individual living cells, we developed a rapid laser temperature stepping method capable of measuring complete thermal melts and kinetic traces in about two min. We find that this method yields improved thermal melts compared to heating a sample chamber or microscope stage. By comparing results for six cells with in vitro data, we show that the protein is stabilized by about 6 kJ/mole in the cytoplasm, but the temperature dependence of folding kinetics is similar to in vitro. The main difference is a slightly steeper temperature dependence of the folding rate in some cells that can be rationalized in terms of temperature-dependent crowding, local viscosity, or hydrophobicity. The observed rate coefficients can be fitted within measurement uncertainty by an effective two-state model, even though PGK folds by a multistate mechanism. We validate the effective two-state model with a three-state free energy landscape of PGK to illustrate that the effective fitting parameters can represent a more complex underlying free energy landscape.


Field longevity of a fluorescent protein marker in an engineered strain of the pink bollworm, Pectinophora gossypiella (Saunders).
The cotton pest, pink bollworm (Pectinophora gossypiella (Saunders)), is a significant pest in most cotton-growing areas around the world. In southwestern USA and northern Mexico, pink bollworm is the target of the sterile insect technique (SIT), which relies on the mass-release of sterile pink bollworm adults to over-flood the wild population and thereby reduce it over time. Sterile moths reared for release are currently marked with a dye provided in their larval diet. There are concerns, however, that this marker fails from time to time, leading to sterile moths being misidentified in monitoring traps as wild moths. This can lead to expensive reactionary releases of sterile moths. We have developed a genetically marked strain that is engineered to express a fluorescent protein, DsRed2, which is easily screened under a specialised microscope. In order to test this marker under field conditions, we placed wild-type and genetically marked moths on traps and placed them in field cages. The moths were then screened, in a double-blind fashion, for DsRed2 fluorescence at regular intervals to determine marker reliability over time. The marker was shown to be robust in very high temperatures and generally proved reliable for a week or longer. More importantly, genotyping of moths on traps by PCR screening of the moths was 100% correct. Our findings indicate that this strain--and fluorescent protein markers in general--could make a valuable contribution to SIT.


Role of IL-17 in Glioma Progression.
There is increasing evidence in the literature pointing to an important role of inflammation during initiation and progression of cancer. Glioblastoma is the most common malignant primary brain tumor with approximately 23,000 newly-diagnosed cases each year in the United States, and has a dismal median survival of only 15 months. Although the blood-brain barrier maintains an immune-privileged status of the brain under steady state, intracranial tumors including gliomas are invariably infiltrated with various types of immune cells. The T helper 17 (Th17) cells, a recently discovered interleukin (IL)-17-producing T cell subtype, have been reported in several extracranial and some intracranial tumors, where they have been implicated in either pro- or antitumor activity depending on the tumor type. Here, we present a succinct review of the current literature on the prevalence and potential role of IL-17 in malignant gliomas. Further mechanistic studies on IL-17 mediated inflammatory pathway in gliomas may provide with opportunities for novel immunotherapeutic interventions.


Comparison of the effects of cycloheximide and inhibin on the gonadotropin subunit messenger ribonucleic acids.
Cycloheximide (CHX) has been shown to mimic the action of inhibin on gonadotropin secretion by pituitary cell cultures. We showed previously that suppression of FSH secretion by inhibin is associated with a rapid and profound suppression of FSH beta mRNA levels. The present study was designed to examine the mechanism of action of CHX and to determine whether inhibin's actions involve new proteins synthesis. Pituitary cell cultures were treated with control medium or medium containing inhibin, CHX, or inhibin plus CHX for 2 or 6 h. At 6 h, secretion of FSH was decreased by inhibin (72% of control), CHX (58% of control), and the combined inhibitors (56% of control). LH secretion was not significantly changed, while that of free alpha-subunit was reduced only by CHX (68% of control). Levels of FSH beta, LH beta, and alpha-subunit mRNAs were measured by Northern analysis. At 2 h inhibin decreased FSH beta mRNA to 49% of the control value. CHX alone had no effect, while CHX plus inhibin produced intermediate levels (77% of control). By 6 h, however, inhibin and CHX each decreased FSH beta mRNA to very low levels (12% and 15% of control, respectively), and in cultures treated with both inhibin and CHX, this RNA was barely detectable. To determine the reversibility of the effects of these inhibitors, cells were incubated with fresh control medium after 6 h. Secretion of FSH and free alpha-subunit remained suppressed 4 h later; recovery was complete by 16 h in inhibin treated cultures. FSH beta mRNA returned to control levels by 4 h in inhibin-treated and by 16 h in CHX-treated cultures. Levels of LH beta and alpha-subunit mRNA were comparable to control values at all times. In conclusion, 1) CHX, like inhibin, suppresses FSH beta mRNA levels, although its actions are less rapid and less rapidly reversible; 2) inhibin requires ongoing protein synthesis for full expression of its inhibitory effects; 3) the synthesis and secretion of LH are much less sensitive to inhibition by either inhibin or CHX than are the synthesis and secretion of FSH; and 4) secretion of free alpha-subunit involves a labile protein(s).


[Noninvasive regional cerebral blood flow measurements by 99mTc-HMPAO using only three-head SPECT system].
A special bed and a new software program were developed for noninvasive regional cerebral blood flow (rCBF) measurements by 99mTc hexamethylpropylene amine oxime (99mTc-HMPAO) using only three-head SPECT system, Siemens MULTI-SPECT3. A large field gamma camera with a minimum of 38 cm-diameter is required for simultaneous acquisition of time activity curves of brain and aortic arch in this noninvasive method. A special bed with short width was designed and inserted obliquely into the gantry of the SPECT system to acquire both time activity curves of brain and aortic arch with the use of diagonal length (51 cm) of a gamma camera with 31 x 41 cm width. Moreover a software program was developed for processing a series of the graphical analysis of time activity curves, Lassen's linearization correction of SPECT images, and drawing of a rCBF map. This noninvasive rCBF measurement using only MULTISPECT3 in a short period with high spatial resolution is quite useful for routine clinical studies.


Ambulatory blood pressure and insulinaemia in different clinical conditions.
The aim of our study was to examine the relation between insulin and ambulatory blood pressure (ABPM) in several clinical conditions. ABPM monitoring was performed with Spacelabs device. Fasting plasma insulin (FPI) was measured by radioimmunoassay. In young, non-obese normotensive subjects (n = 32) there were correlations between FPI and both asleep (r = 0.61, p < 0.001), and awake systolic ABPM (r = 0.44, p < 0.01). We have not observed any significant correlation between FPI and ABP in borderline hypertensives or in patients with established hypertension. In a group of 21 hypertensive type 2 diabetics (age 52 +/- 8 years) there was no significant correlation between FPI and ABPM. Among 14 normotensive type 1 diabetics (age 31 +/- 7 years, diabetes duration > 10 years) there was a significant negative correlation between the daily dose of insulin and 24-h systolic ABPM (r =-0.63, p < 0.02). In 20 patients with renal failure on chronic haemodialysis we have found a significant negative correlation between FPI and 24-h systolic APBM (r = 0.80, p < 0.001) and 24-h diastolic ABPM (r = -0.55, p < 0.05). Similar negative correlations were found in 20 nondialysed subjects with moderate chronic renal failure. Taken together, our results suggest that insulin could reveal its hypertensive or vasodilatory effect which depends on a clinical condition of the studied subjects.


Effect of sintering process on color parameters of nano-sized yttria partially stabilized tetragonal monolithic zirconia.
Sintering process is responsible for aesthetic of zirconia restoration. This study evaluated the effect of different sintering temperatures and sintered-holding times on color parameters of monolithic zirconia. One hundred and thirty five zirconia bar specimens (width-length-thickness = 10×20×1.5 mm) were prepared from yttria-stabilized tetragonal zirconia polycrystalline (Y-TZP) ceramic and randomly divided into nine groups to be sintered at different temperatures [decreasing- (SD, 1350°C), regular- (SR, 1450°C), and increasing- (SI, 1550°C) sintering temperature] and different sintered-holding times [shortening- (HS, 60 min), regular- (HR, 120 min), and prolonged- (HP, 180 min) sintered-holding time]. Color appearance (ΔE), translucency parameter (TP), contrast ratio (CR), and opalescence parameter (OP) were determined with spectrophotometer. An analysis of variance (ANOVA) and Tukey's multiple comparisons were used to determine for statistically significant difference of color parameters (α=0.05). Crystal sizes were microscopically examined using scanning electron microscope (SEM), and phase composition of zirconia was determined using X-ray diffraction (XRD). The mean±sd for ΔE, TP, CR, OP were 82.28±1.27, 1.4±0.13, 0.982±0.004, 1.25±0.15 for SDHS, 78.38±0.74, 2.16±0.10, 0.967±0.005, 1.90±0.11 for SDHR, 74.43±0.91, 2.24±0.10, 0.964±0.004, 1.94±0.09 for SDHP, 76.31±1.22, 3.03±0.10, 0.945±0.003, 2,50±0.09 for SRHS, 74.51±1.27, 3.19±0.17, 0.942±0.003, 2.65±0.16 for SRHR, 73.94±0.49, 3.42±0.10, 0.937±0.003, 2,83±0.09 for SRHP, 76.30±0.43, 3.16±0.09, 0.937±0.002, 2.48±0.09 for SIHS 76.73±1.15, 3.05±0.20, 0.939±0.005, 2.38±0.17 for SIHR, and 75.32±1.37, 2.95±0.18, 0.942±0.006, 2.33±0.15 for SIHP. The ΔE, TP, CR, and OP were significantly affected by altering sintering temperatures and holding times (p<0.05). Increasing sintering temperature and extending sintering time significantly improved color appearance, translucency, contrast, and opalescence of Y-TZP (p<0.05) as evidenced by enlarging grain size and increasing t→m phase shift. Raising sintering temperature and prolonging sintering time lead to better color appearance, translucency, contrast and opalescence of nano-sized monolithic Y-TZP, and are suggested for sintering process. Key words:Color appearance, contrast, monolithic zirconia, opalescence, sintering process, translucency.


Life cycle assessment of microgreen production: effects of indoor vertical farm management on yield and environmental performance.
The global production of plant-based foods is a significant contributor to greenhouse gas emissions. Indoor vertical farms (IVFs) have emerged as a promising approach to urban agriculture. However, their environmental performance is not well understood, particularly in relation to operational choices where global warming potentials (GWP) can vary between 0.01-54 kg CO2e/kg-1 of leafy greens produced. We conducted a life cycle assessment (LCA) of a building-integrated IVF for microgreen production to analyse a range of operational conditions for cultivation: air temperature, CO2 concentration, and photoperiod. We analyzed a dynamic LCA inventory that combined a process-based plant growth model and a mass balance model for air and heat exchange between the chamber and the outside. Results showed that the GWP of IVFs can vary greatly depending on the operation conditions set, ranging from 3.3 to 63.3 kg CO2e/kg-1. The optimal conditions for minimizing GWP were identified as 20 ℃, maximum CO2 concentration in the chamber, and maximum photoperiod, which led to a minimum GWP of 3.3 kg CO2e/kg-1 and maximum production of 290.5 kg fresh weight week-1. Intensification of production thus led to lower impacts because the marginal increase in yield due to increased resource use was larger than the marginal increase in impact. Therefore, adjusting growing conditions is essential for the sustainability of urban food production.


Impaired beta-cell and alpha-cell function in African-American children with type 2 diabetes mellitus--"Flatbush diabetes".
The incidence of type 2 diabetes mellitus (T2DM) in children and adolescents has substantially increased over the past decade. This is attributed to obesity, insulin resistance and deficient beta-cell function. In children a pubertal increase in insulin resistance and an inability to mount an adequate beta-cell insulin response results in hyperglycemia. Adults with T2DM have a diminished first phase response to intravenous glucose and a delayed early insulin response to oral glucose. Long-term studies show progressive loss of beta-cell function in T2DM in adults; however, such long-term studies are not available in children. To characterize beta- and alpha-cell function in African-American adolescents with established T2DM, we used mixed meal, intravenous glucagon and oral glucose tolerance testing and compared them to obese non-diabetic controls. T2DM was defined as fasting C-peptide >0.232 nmol/l and absent autoimmune markers. BETA-CELL FUNCTION: Meal testing in 24 children and adolescents with T2DM, mean age 14 years, BMI 30 kg/m2, Tanner stage II-V, HbA1c 8.9%, were compared with BMI- and age-matched controls. Forty percent presented with DKA. Half were treated with insulin and half with diet/oral anti-diabetic agents. Although absolute C-peptide response in both groups was similar, the incremental rise in C-peptide relative to plasma glucose in the patients with T2DM compared to controls was 40% and 35% lower 30 and 60 min after the meal, p <0.007 and p <0.026. Glucagon testing in 20 pediatric patients with T2DM compared with 15 matched controls showed significantly lower 6 min stimulated C-peptide relative to the ambient plasma glucose in patients with T2DM compared to controls (0.039 +/- 0.026 vs 0.062 +/- 0.033, p <0.05). The clinical utility is that 78% of patients with a 6 min C-peptide <1.4 nmol required insulin, while 81% of those >1.4 nmol required oral anti-diabetic agents, p <0.0001. Furthermore, the duration of T2DM up to 5 years after diagnosis was associated with lower fasting and glucagon-stimulated C-peptide levels, implying worsening beta-cell function over time, even in children and adolescents. ALPHA-CELL FUNCTION: During meal testing, children and adolescents with T2DM had less suppression of plasma glucagon than non-diabetic controls; this was more severe with longer duration of T2DM and poorer glycemic control. BETA-CELL RECOVERY: In African-American and Hispanic adults, intensive treatment of blood glucose may achieve beta-cell recovery with 35-40% of newly diagnosed patients going into remission after 6 months treatment. They remain off anti-diabetic pharmacological agents in remission for a median of over 3 years with normal HbA1c levels. We hypothesize this to be due to removal of a critical component of glucose or lipotoxicity at the level of the beta-cell and/or peripheral tissue. Four of 20 African-American children presenting with mean glucose 650 mg/dl maintained normal HbA1c levels on small doses of metformin after initial treatment with multiple insulin injections with or without metformin. This suggests a marked recovery of beta-cell function, similar to that in adults. T2DM in children, as in adults, is characterized by insulin deficiency relative to insulin resistance. Plasma C-peptide levels may be clinically useful in guiding therapeutic choices, since patients with lower levels required insulin treatment; beta-cell function is also diminished with longer duration of T2DM. The possibility exists that in children, as in adults, intensive glycemic regulation may allow for beta-cell recovery and preservation. Thus, optimum beta- and alpha-cell function are central to the prevention of DM and maintenance of good glycemic control in African-American and Hispanic children and adolescents with T2DM.


Combined modifications of mexiletine pharmacophores for new lead blockers of Na(v)1.4 channels.
Previously identified potent and/or use-dependent mexiletine (Mex) analogs were used as template for the rational design of new Na(v)-channel blockers. The effects of the novel analogs were tested on sodium currents of native myofibers. Data and molecular modeling show that increasing basicity and optimal alkyl chain length enhance use-dependent block. This was demonstrated by replacing the amino group with a more basic guanidine one while maintaining a proper distance between positive charge and aromatic ring (Me13) or with homologs having the chirality center nearby the amino group or the aromatic ring. Accordingly, a phenyl group on the asymmetric center in the homologated alkyl chain (Me12), leads to a further increase of use-dependent behavior versus the phenyl Mex derivative Me4. A fluorine atom in paraposition and one ortho-methyl group on the xylyloxy ring (Me15) increase potency and stereoselectivity versus Me4. Charge delocalization and greater flexibility of Me15 may increase its affinity for Tyr residues influencing steric drug interaction with the primary Phe residue of the binding site. Me12 and Me15 show limited selectivity against Na(v)-isoforms, possibly due to the highly conserved binding site on Na(v). To our knowledge, the new compounds are the most potent Mex-like Na(v) blockers obtained to date and deserve further investigation.


Abscopal effect with fever of unknown cause during radiotherapy: Two case reports and review of the literature.
The abscopal effect is a rare phenomenon that is defined as regression of tumor lesions distant from irradiation targets. At our department, two cases with an abscopal effect with fever of unknown cause (FUC) and an inflammatory response during radiotherapy were encountered. Radiotherapy is a local treatment; therefore, it rarely causes systemic side effects during radiotherapy, and if a patient develops a fever during radiotherapy, it is frequently considered tumor fever. We experienced 2 cases of FUC during irradiation followed by abscopal effect. The obvious relationship between the abscopal effect and the fever remains to be clarified. However, FUC during radiotherapy may be a hint to the abscopal effect, considering that immune response and cytokines are closely related to the abscopal effect.


Mega-abrasion: Case History Report on Management of a Fluorosis-Affected Patient.
Different approaches ranging from veneers and crowns to employing the more conservative use of freehand resin composites have been proposed to correct esthetic problems caused by enamel fluorosis (EF). However, the protocol of mega-abrasion followed by microabrasion may be used as an alternative. It is less invasive and can minimize the unesthetic brown-to-whitish spots caused by EF. This article describes an easy and low-cost technique for managing enamel defects in a patient with severe enamel fluorosis.


Shifting preferences for choice-making opportunities through histories of differential reinforcer quality.
Children of both typical and atypical cognitive development tend to prefer contexts in which their behavior results in a choice of reinforcers rather than a single reinforcer, even when the reinforcer accessed is identical across conditions. The origin of this preference has been attributed speculatively to behavioral histories in which choice making tends to be associated with differentially beneficial outcomes. Few studies have evaluated this claim, and those that have, have yielded mixed results. We provided five preschool-aged children experiences in which choice-making and no-choice contexts were differentially associated with higher preference and larger magnitude reinforcers, and we assessed changes in their preference for choice and no-choice contexts in which outcomes were equated. These conditioning experiences resulted in consistent and replicable shifts in child preference, indicating that preference for choice is malleable through experience.


Mucus Hypersecretion in Chronic Obstructive Pulmonary Disease and Its Treatment.
Most patients diagnosed with chronic obstructive pulmonary disease (COPD) present with hallmark features of airway mucus hypersecretion, including cough and expectoration. Airway mucus function as a native immune system of the lung that severs to trap particulate matter and pathogens and allows them to clear from the lung via cough and ciliary transport. Chronic mucus hypersecretion (CMH) is the main factor contributing to the increased risk of morbidity and mortality in specific subsets of COPD patients. It is, therefore, primarily important to develop medications that suppress mucus hypersecretions in these patients. Although there have been some advances in COPD treatment, more work remains to be done to better understand the mechanism underlying airway mucus hypersecretion and seek more effective treatments. This review article discusses the structure and significance of mucus in the lungs focusing on gel-forming mucins and the impacts of CMH in the lungs. Furthermore, we summarize the article with pharmacological and nonpharmacological treatments as well as novel and interventional procedures to control CMH in COPD patients.


Maternal HBsAg status and infant size--a Faustian bargain?
Information on the impact of maternal hepatitis B virus (HBV) infection on pregnancy outcome is conflicting. Some studies reported an association with increased infant birthweight, which could be interpreted as advantageous to pregnancy. A retrospective study was performed to compare birthweight outcome between 6261 and 55,817 singleton pregnancies in mothers screened positive and negative for hepatitis B surface antigen (HBsAg), respectively. The HBsAg positive women were younger, had higher body mass index (BMI) and incidence of overweight, but less gestational weight gain, and were associated with increased macrosomia (birthweight ≥4000 g) in mothers <35 years (odds ratio, OR, 1.28), BMI ≥25 kg/m(2) (OR 1.24), without gestational diabetes mellitus (GDM, OR 1.19), and in male infants (OR 1.18). It was also associated with increased large-for-gestational age (LGA, birthweight >90th percentile) infants in nulliparas (OR 1.13), age <35 years (OR 1.12), BMI ≥25 kg/m(2) (OR 1.19), with (OR 1.36) and without (OR 1.09) GDM, and in male infants (OR 1.13). When the effects of high BMI, advanced age, GDM, and male infants were controlled for, positive HBsAg was significantly associated with macrosomic (adjusted odds ratio, aOR, 1.15) and LGA (aOR 1.11) infants. In view of the latest findings on the association between high infant birthweight with increased risk of obesity, diabetes mellitus, and various forms of malignancies from childhood to adulthood, further studies are warranted to determine if maternal hepatitis B infection would impact adversely on the long-term health of the offspring through its effect on increasing birthweight.


Serum resistin: A possible link between inflammation, hypertension and coronary artery disease.
Inflammation is considered as the main triggering factor in evolution of atherosclerotic pathology of heart and blood vessels. Resistin, an inflammatory cytokine is proved to be a main mediator of initiation and progression of mechanisms leading to atherosclerosis, hypertension and ultimately to coronary artery disease. Our objective was to compare the levels of serum resistin, C-reactive protein and total leucocyte count in subjects of hypertension and coronary artery disease; and to observe the correlation of serum resistin with CRP and TLC in the study participants. Eighty selected participants were divided into four equal groups including normal healthy participants, newly diagnosed cases of hypertension, stable angina pectoris and myocardial infarction, both with hypertension. The study was conducted in the physiology department of Post Graduate Medical Institute Lahore, during 2013. After consent, history and examination, fasting blood samples of the participants were collected. Serum resistin and C-reactive protein were determined by using standard techniques of enzyme linked immunosorbent assay, while total leukocyte count by automated hematology analyzer. The values of serum resistin, C- reactive protein and total leukocyte count were found significantly raised in patients of hypertension, angina pectoris and myocardial infarction with hypertension as compared to normal participants (p<0.001 for all). Significantly positive correlation of resistin was observed with TLC only in hypertensive patients of myocardial infarction (r = 0.459, n = 20, p = 0.042) while in other study groups correlation between resistin and TLC as well as CRP was non-significant. Serum resistin levels along with CRP and TLC are significantly raised in patients of hypertension and coronary artery disease while resistin levels revealed significantly positive correlation with TLC in hypertensive patients of myocardial infarction.


Probability Theory Plus Noise: Descriptive Estimation and Inferential Judgment.
We describe a computational model of two central aspects of people's probabilistic reasoning: descriptive probability estimation and inferential probability judgment. This model assumes that people's reasoning follows standard frequentist probability theory, but it is subject to random noise. This random noise has a regressive effect in descriptive probability estimation, moving probability estimates away from normative probabilities and toward the center of the probability scale. This random noise has an anti-regressive effect in inferential judgement, however. These regressive and anti-regressive effects explain various reliable and systematic biases seen in people's descriptive probability estimation and inferential probability judgment. This model predicts that these contrary effects will tend to cancel out in tasks that involve both descriptive estimation and inferential judgement, leading to unbiased responses in those tasks. We test this model by applying it to one such task, described by Gallistel et al. ). Participants' median responses in this task were unbiased, agreeing with normative probability theory over the full range of responses. Our model captures the pattern of unbiased responses in this task, while simultaneously explaining systematic biases away from normatively correct probabilities seen in other tasks.


Parents' perception of their children's development, perception of satisfaction, social support and level of depression.
The purpose of this study was to describe how parents and guardians in rural South Africa reported on the development of their children, their perception of social support and satisfaction, their level of depression, and the relationships that exist among the variables. The data from a larger research study were used to answer these questions: Is there a relationship between the child's development status and the parents/guardians level of satisfaction and social support? Is there a difference in the educational level of the parents/guardians and their perception of social support and the level of satisfaction? What is the percentage of parents/guardians working/not working? Is there a relationship between the status of work of the parents/guardians and the level of depression? Is there a relationship between the age of the parents/guardians and their level of depression? A convenience sample of 198 parents/guardians was recruited. Interviews were conducted using a structured interview guide. Relationships and differences were analyzed using Pearson's r correlation index, paired t-tests and regression analysis. The majority (73.7%) of the respondents were unemployed. No significant relationships were found between the child's developmental status and the parents/guardians perceived level of social support and satisfaction. However, the difference between the educational level of parents/guardians and the level of perceived social support was significant. Additionally, as the amount of family strains increased so did the parental level of depression.


[Methods and clinical values for minimal residual disease detection in patients with multiple myeloma].
The complete response (CR) rates in patients with multiple myeloma (MM) have been rapidly increasing after the approval of novel agents such as proteasome inhibitors, immunomodulatory drugs, histone deacetylase inhibitors, and monoclonal antibodies. Recently, patients administered with a combination of novel agents with autologous stem-cell transplantation achieved a 70% CR rate. The assessment of minimal residual disease (MRD) is necessary for effective stratification of the CR cases and the generation of accurate prognoses. This review summarizes recent topics pertaining to MRD detection methods (multiparameter flow cytometry, allele-specific oligonucleotide real-time quantitative polymerase chain reaction, droplet digital PCR, and next-generation sequencing) and the prognostic value of MRD assessment in patients with MM.


Characterization and Comparative Analysis of MicroRNAs in 3 Representative Red Algae.
MicroRNA (miRNA) is a key regulator at the gene posttranscriptional regulation level. We have previously identified miRNAs and their putative targets in 3 representative red algae, Chondrus crispus, Galdieria sulphurariais and Porphyridium purpureum. In this study, unique molecular and evolutionary characterization of miRNAs were revealed in the 3 red algae based on the comparative miRNAs profiling. Genome locations of small RNAs (sRNAs), miRNAs and MIRNAs (MIRs) in the 3 red algae were shown by collinearity analysis. Characterization of miRNAs and MIRs were profiled via bioinformatics analysis. Taken MIR156s and miR156s for examples, red algae miRNAs evolutionary features were demonstrated via phylogenetic and evolutionary information analysis. MiRNA targets main inhibition type was validated via performing data statistics and RLM-RACE PCR. Key target genes and their function were predicted by the common Gene Ontolgoy (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Quantity, nucleotide bias and common sequences of miRNAs were analyzed in the 3 red algae. Four typical precursor structures and primary molecular features of red algae miRNAs were profiled. Genome-wide collinearity analysis of sRNAs, miRNAs and MIRs in the 3 red algae was performed to show their distribution and interrelation based on the deep sequencing data. Taken red algae MIR156s for example, their family members and sequences divergence were demonstrated. The whole evolutionary processes of miR156s and pre-miR156s in red algae were steady with negative selected pressure though diverse phylogenetic relationships and evolutionary parameters showed. Through 3 red algae miR156 targets validation, cleavage was validated as their main miRNA targets inhibition type. The common target genes (GO:0009536) enriched significantly for plastid formation will provide important insights for red algal biopigment research. The common KEGG pathways (ko01100) enriched significantly were predicted without a detailed reference metabolic map. MiRNA plays an essential role in gene expression regulation involved in diverse biological processes of red algae. Comprehensive molecular and evolutionary features of miRNAs in the 3 red algae will provide insights for further utilizing the algae resources at the molecular level.


Synthesis and two-photon absorption enhancement of porphyrin macrocycles.
Improving the nonlinear optical properties of organic materials through transition to macromolecular architectures of highly conjugated chromophores has been shown to be a viable strategy for generating materials suitable for TPA applications. In this study we display a simple and elegant method by which to synthesize macrocycles of porphyrin dimers. Two-photon absorption studies show 2 orders of magnitude enhancement of cross-section of the material, giving a maximum delta(2) of 10(6) GM.


[Triple pelvic osteotomy (according to Tönnis and Kalchschmidt) in the treatment of acetabular dysplasia--medium-term results].
The triple pelvic osteotomy according to Tönnis and Kalchschmidt has gained major significance in the surgical treatment of acetabular dysplasia in adolescents and adults. The curative and prophylactic potentials of this operative procedure are reviewed in this study by means of a clinical and radiological follow-up trial. 34 patients/38 hip joints treated by the triple pelvic osteotomy between 1996 and 2005 were reviewed clinically and radiologically with an average follow-up of 3.5 years. The radiological classification into grades of deviation was done according to Tschauner et al., clinically the preoperative Harris hip score was determined, as well as common epidemiologic parameters. At follow-up, the preoperatively determined radiological and clinical parameters were controlled and subjected to statistical analysis using Student's t-test for the comparison of means between two paired groups. Finally, the subjective assessment of the operation's result by the patients was registered. A statistically highly significant amelioration of the Harris hip score, relevant radiological angles and grades of deviation could be demonstrated at follow-up. This fact coincided with a high or very high patient satisfaction in 81.7% of the cases. The conversion rate to alloarthroplasty was 2.6%. The complication profile was within the scope of similar elective hip surgeries. The triple pelvic osteotomy according to Tönnis and Kalchschmidt has shown in the present study a high and constant potential with regard to reduction/absence of disorders as well as a statistically highly significant amelioration of clinical scores and relevant radiological angles. In our experience, this surgical method can be regarded as the joint-preserving method of choice in the cardinal indication of adolescent and adult acetabular dysplasia.


Ruthenium complexes/polypeptide self-assembled nanoparticles for identification of bacterial infection and targeted antibacterial research.
Bacterial infection has been a threat to human health, and so early diagnosis and treatment of bacterial infection is an urgent problem that needs to be solved. In this work, a multifunctional theranostic selenium nanoplatform (Se@PEP-Ru NPs) with early imaging diagnosis and efficient treatment of bacterial infections was designed and constructed. First, the antibacterial peptide UBI29-41 (PEP) was linked to functionalized Selenium nanoparticles (NPs), which enhanced the stability of the antimicrobial peptide and also caused the nanocomposites to specifically target bacterial infection. Ruthenium complexes with good antibacterial activity and fluorescence properties were then coated on to their outer layers. It was worth mentioning that, when the resulting nanoprobe was injected into mice by intravenous injection it was found to be sensitive to sites of bacterial infection for selective fluorescence imaging and targeted therapy. Thus, it can be used to distinguish between bacterial infection, inflammation, and tumor-induced tissue infection with high specificity. In the further antibacterial activity experiments, Ruthenium complexes showed synergistic antimicrobial activity with Se NPs, which indicated that the antibacterial activity of Se@PEP-Ru NPs was the strongest that could promote wound healing. Thus, Se@PEP-Ru NPs appears to be a promising antimicrobial with good biocompatibility, excellent selectivity, and potent antimicrobial activity.


Healing of freeze-treated hemimaxillary bone.
Earlier studies suggested that the combined techniques of excision, freezing, and reimplantation of mandibular bone may provide a new method for the treatment of intraoral bone tumors. An experiment was conducted to investigate the bone healing and revascularization associated with resected, frozen, and reimplanted maxillary bone. In all the FT and NFT hemimaxillas evaluated, complete reconstitution occurred. The degree of revascularization was similar in both FT and NFT monkeys, suggesting that freezing does not adversely effect subsequent revascularization. Compared to the sham operated sides, both FT and NFT hemimaxillas showed a high degree of revascularization; this phenomenon was greatest at six weeks postoperatively. Microscopically, in all the animals, periosteal osteoblastic activity was present in variable amounts. The interstitial tissue was predominantly fibrous rather than fatty marrow, and no detectable differences were noticed between FT and NFT animals in the same time interval.


The pharmacodynamics and pharmacokinetics of the metabolite 3-desacetylvecuronium (ORG 7268) and its parent compound, vecuronium, in human volunteers.
The pharmacology of 3-desacetylvecuronium, the principal metabolite of vecuronium, was investigated. We studied 12 healthy volunteers, each on two occasions. First they received 3-desacetylvecuronium alone and then, on a later occasion, vecuronium. Six subjects received a large dose of each drug (pharmacokinetic study), the remaining six received a small dose (pharmacodynamic study). Drug concentrations in plasma and urine were measured using capillary gas chromatography. Neuromuscular block was assessed by measuring force of contraction of the adductor pollicis. Drug plasma concentration vs. time and neuromuscular effect data were analyzed by nonlinear mixed-effects modeling. 3-Desacetylvecuronium, compared with vecuronium (median, range in parentheses), had a smaller plasma clearance, 3.51 (2.11-6.57) vs. 5.39 (5.04-7.19) ml.kg-1.min-1; a larger steady-state distribution volume, 254 (215-410) vs. 152 (111-170) ml.kg-1; a longer terminal elimination half-life 116 (44-672) vs. 34 (25-61) min and a longer mean residence time, 67 (42-145) vs. 26 (18-32) min (P < .05). Renal clearances of 3-desacetylvecuronium and vecuronium were 0.85 (0.15-1.24) and 0.58 (0.16-0.66) ml.kg-1.min-1, respectively (P < .05). Conversion to 3-desacetylvecuronium accounted for 12% of vecuronium's clearance. Concentrations of 3-desacetylvecuronium and vecuronium that produced 50% neuromuscular block were 123 (109-154) and 102 (71-123) ng.ml-1, respectively (P < .05). 3-Desacetylvecuronium is a potent neuromuscular blocking drug and may be responsible for episodes of prolonged paralysis after long-term administration of vecuronium to patients in intensive care units.


Considering protonation as a posttranslational modification regulating protein structure and function.
Posttranslational modification is an evolutionarily conserved mechanism for regulating protein activity, binding affinity, and stability. Compared with established posttranslational modifications such as phosphorylation or ubiquitination, posttranslational modification by protons within physiological pH ranges is a less recognized mechanism for regulating protein function. By changing the charge of amino acid side chains, posttranslational modification by protons can drive dynamic changes in protein conformation and function. Addition and removal of a proton is rapid and reversible and, in contrast to most other posttranslational modifications, does not require an enzyme. Signaling specificity is achieved by only a minority of sites in proteins titrating within the physiological pH range. Here, we examine the structural mechanisms and functional consequences of proton posttranslational modification of pH-sensing proteins regulating different cellular processes.


Recent Molecular Aspects and Integrated Omics Strategies for Understanding the Abiotic Stress Tolerance of Rice.
Rice is an important staple food crop for over half of the world's population. However, abiotic stresses seriously threaten rice yield improvement and sustainable production. Breeding and planting rice varieties with high environmental stress tolerance are the most cost-effective, safe, healthy, and environmentally friendly strategies. In-depth research on the molecular mechanism of rice plants in response to different stresses can provide an important theoretical basis for breeding rice varieties with higher stress resistance. This review presents the molecular mechanisms and the effects of various abiotic stresses on rice growth and development and explains the signal perception mode and transduction pathways. Meanwhile, the regulatory mechanisms of critical transcription factors in regulating gene expression and important downstream factors in coordinating stress tolerance are outlined. Finally, the utilization of omics approaches to retrieve hub genes and an outlook on future research are prospected, focusing on the regulatory mechanisms of multi-signaling network modules and sustainable rice production.


Visualisation of the left anterior descending coronary artery on CT images used for breast radiotherapy planning.
To assess the visualisation of the left anterior descending (LAD) coronary artery on CT images used for breast radiation treatment planning. Delineation of the LAD artery was achieved for 25 breast patients by 1 radiologist and 1 radiation oncologist independently on two sets of images for each patient: one pre-operative CT scan using intravenous (IV) contrast media to determine the primary gross tumour volume (GTV) and one post-operative CT scan used for treatment planning. A Student's paired t-test was used to compare the number of CT slices in which the LAD was visible for each patient in the two series. Interpolations and extrapolations of the LAD volume were performed for the left-sided cases using a published heart atlas in order to report doses to the LAD structure. There was a non-significant difference between the results with and without IV contrast media (p=0.34 for the radiologist; p=0.90 for the radiation oncologist). The visible LAD artery corresponded to a 30% portion (range 12-47%) of the interpolated structure. The maximum dose to the left artery varied widely, from 2.7 to 41.7 Gy, in the group of patients with left breast tumours. The largest values (>25 Gy) corresponded to those patients in whom the LAD artery distal extremity lay inside the breast fields. With the current planning CT protocol, only one-third of the LAD artery could be objectively visualised. Contrast-enhanced imaging used for GTV delineation before the breast surgery did not improve the visualisation of the artery. This study has revealed the lack of consistency that may be encountered when contouring heart vessels, thereby questioning the reliability of dose reporting.


On-filter direct amplification of Legionella pneumophila for rapid assessment of its abundance and viability.
Guidelines and regulations to control Legionella pneumophila in cooling water systems of large buildings are evolving due to the increasing number of outbreaks. Rapid, on-site, simple, and sensitive quantification methods that are also able to assess viability may be extremely useful in monitoring and control. Culture-based methods for measuring L. pneumophila may take 4-10 days and qPCR-based methods are also slow, requiring at least a day from sample to result, albeit mainly due to the need for sample transport to a centralized laboratory. This study reports a rapid isothermal amplification method for L. pneumophila concentration and detection with live/dead differentiation under field conditions. Using an on-filter direct amplification (i.e., amplification of cells without DNA extraction and purification) approach with propidium monoazide (PMA), and a real time isothermal amplification platform (Gene-Z), L. pneumophila could be detected in 1-2 h at ∼1 cfu/100 ml of tap water. Signature sequences from 16S rRNA and cadA genes were used as genetic markers for L. pneumophila and loop-mediated isothermal amplification (LAMP) primers were designed using Primer Explorer V4. Result were also compared with direct amplification of cells spiked into distilled, tap, and cooling water samples as well as extracted DNA by qPCR. This method may be useful to managers of cooling water systems in large buildings for rapid detection of L. pneumophila. The overall approach of on-site sample concentration, on-filter amplification, and live/dead differentiation may be extended to other organisms where analytical sensitivity and speed are equally important.


Non-communicable diseases (NCDs) in developing countries: a symposium report.
In recent years, non-communicable diseases (NCDs) have globally shown increasing impact on health status in populations with disproportionately higher rates in developing countries. NCDs are the leading cause of mortality worldwide and a serious public health threat to developing countries. Recognizing the importance and urgency of the issue, a one-day symposium was organized on NCDs in Developing Countries by the CIHLMU Center for International Health, Ludwig-Maximilians-Universität, Munich on 22nd March 2014. The objective of the symposium was to understand the current situation of different NCDs public health programs and the current trends in NCDs research and policy, promote exchange of ideas, encourage scientific debate and foster networking, partnerships and opportunities among experts from different clinical, research, and policy fields. The symposium was attended by more than seventy participants representing scientists, physicians, academics and students from several institutes in Germany and abroad. Seven key note presentations were made at the symposium by experts from Germany, UK, France, Bangladesh and Vietnam. This paper highlights the presentations and discussions during the symposium on different aspects of NCDs in developing countries. The symposium elucidated the dynamics of NCDs in developing countries and invited the participants to learn about evidence-based practices and policies for prevention and management of major NCDs and to debate the way forward.


Circulating long noncoding RNA DKFZP434I0714 predicts adverse cardiovascular outcomes in patients with end-stage renal disease.
Cardiovascular (CV) diseases are major causes of mortality in uremic patients. Conventional risk factors fail to identify uremic patients with increased propensity for adverse CV outcomes. We aimed to test the hypothesis that circulating long noncoding RNAs (lncRNAs) could be a prognostic marker to predict adverse CV outcomes in uremic patients. Plasma lncRNAs were profiled in patients with end-stage renal disease (ESRD, n = 28) or chronic kidney disease (CKD, n = 8) and in healthy (n = 12) subjects by RNA sequencing. A total of 179 lncRNAs were significantly dysregulated with ESRD; the expression signature of plasma lncRNAs distinguished ESRD from both CKD and control samples. Analysis on a microarray dataset obtained from renal biopsy samples of patients with advanced kidney disease (GSE66494) revealed that a significant proportion of plasma lncRNAs (30.7%) and mRNAs (49.5%) dysregulated with uremia were similarly dysregulated in diseased kidneys, suggesting that plasma RNA profiles mirror the transcriptomal changes in diseased kidney tissues. Further analyses identified eight plasma lncRNAs as potential predictors of adverse CV outcomes in uremic patients. Validation study in an independent cohort of ESRD patients (n = 111) confirmed that elevated plasma lncRNA DKFZP434I0714 is a significant independent predictor of adverse CV outcomes in uremic patients. Additional experiments demonstrated the functional involvement of DKFZP434I0714 in the pathogenesis of endothelial dysfunction. In summary, plasma lncRNA expression signature reflects the disease states of uremia. Elevated plasma levels of lncRNA DKFZP434I0714 in uremic patients portend a worse CV outcome and warrant closer monitoring and more aggressive management.


Correlation between the distribution of smooth muscle or non muscle myosins and alpha-smooth muscle actin in normal and pathological soft tissues.
The distribution of smooth muscle (SM) and non muscle myosins was compared with that of alpha-SM actin in various normal and pathological tissues and in cultured cells by means of indirect immunofluorescence using a monoclonal antibody specific for alpha-SM actin [anti-alpha sm-1, Skalli et al., 1986b] and two polyclonal antibodies raised against bovine aortic myosin (ABAM) and human platelet myosin (AHPM), respectively. In normal tissues ABAM stained vascular and parenchymal smooth muscle cells (SMC), myoepithelial cells and myoid cells of the testis in a pattern similar to that reported by other authors with antisera raised against non vascular SM myosin. Cells stained with ABAM were always positive for anti-alpha sm-1. In human and experimental atheromatous plaques, most cells were positive for AHPM; a variable proportion was also stained for ABAM plus anti-alpha sm-1. Myofibroblasts from rat granulation tissue, Dupuytren's nodule and stroma from breast carcinoma were constantly positive for AHPM and negative for ABAM; however, myofibroblasts from Dupuytren's nodule and breast carcinoma were anti-alpha sm-1 positive. Early primary cultures of rat aortic SMC were positive for ABAM and anti-alpha sm-1 and became negative for ABAM and positive for AHPM after a few days in culture. They remained positive for AHPM and anti-alpha sm-1 after passages; the staining of AHPM and anti-alpha sm-1 appeared to be colocalized along the same stress fibers. These results may be relevant for the understanding of SMC function and adaptation, and show that in non malignant SMC proliferation, alpha-SM actin represents a more general marker of SM origin than SM myosin.


Secondary mAb--vcMMAE conjugates are highly sensitive reporters of antibody internalization via the lysosome pathway.
Monoclonal antibodies (mAb) selectively recognizing tumor surface antigens are an important and evolving approach to targeted cancer therapy. One application of therapeutic mAbs is drug targeting via mAb-drug conjugate (ADC) technology. Identification of mAbs capable of internalizing following antigen binding has been accomplished by tracking decline of surface-bound mAb or by internalization of a secondary mAb linked to a toxin. These methods may not be sufficiently sensitive for screening nor wholly predictive of the mAbs' capacity for a specific drug delivery. We have developed a highly selective and sensitive method to detect mAbs for cell internalization and drug delivery. This system uses secondary anti-human or anti-murine mAbs conjugated to the high-potency drug monomethyl auristatin E (MMAE) via a highly stable, enzymatically cleavable linker. Prior studies of this drug linker technology demonstrated internalization of a primary ADC leads to trafficking to lysosomes, drug release by lysosomal cathepsin B, and ensuing cell death. A secondary antibody--drug conjugate (2 degrees ADC) capable of binding primary mAbs bound to the surface of antigen-positive cells has comparable drug delivery capability. The system is sufficiently sensitive to detect internalizing mAbs in nonclonal hybridoma supernatants and is predictive of the activity of subsequently produced primary ADC. Because of their high extracellular stability, the noninternalized 2 degrees ADC are 100--1000-fold less toxic to cells over extended periods of time, permitting an assay in which components can be added without need for separate wash steps. This homogeneous screening system is amenable to medium-throughput screening applications and enables the early identification of mAbs capable of intracellular trafficking for drug delivery and release.


Effects of dietary manganese deficiency on rat pancreatic amylase mRNA levels.
Manganese deficiency is associated with increased pancreatic amylase activity in rats. The present study investigated whether this increase in amylase activity is associated with increased pancreatic amylase messenger RNA (mRNA) levels. Weanling rats were fed a high carbohydrate diet containing either 40 micrograms Mn/g (control) or 0.5 microgram Mn/g (deficient) for 4 to 8 wk. Mn deficiency was confirmed by determining hepatic Mn content, which was significantly lower in Mn-deficient rats than in controls. Pancreatic RNA from both groups of rats was hybridized with 32P-labeled complementary DNAs for amylase and trypsinogen. Amylase mRNA levels were increased in rats fed Mn-deficient diets for both 4 wk (200%) and 8 wk (250%) when compared with respective control levels. In contrast, Mn deficiency was not associated with alterations in trypsinogen mRNA levels. Serum levels of insulin and corticosterone, hormones known to increase pancreatic amylase mRNA, were not affected by Mn deficiency. These observations suggest that Mn may participate in the regulation of amylase gene expression in a manner that is independent of insulin and corticosterone.


Human umbilical cord blood-borne fibroblasts contain marrow niche precursors that form a bone/marrow organoid in vivo.
Human umbilical cord blood (CB) has attracted much attention as a reservoir for functional hematopoietic stem and progenitor cells, and, recently, as a source of blood-borne fibroblasts (CB-BFs). Previously, we demonstrated that bone marrow stromal cell (BMSC) and CB-BF pellet cultures make cartilage in vitro Furthermore, upon in vivo transplantation, BMSC pellets remodelled into miniature bone/marrow organoids. Using this in vivo model, we asked whether CB-BF populations that express characteristics of the hematopoietic stem cell (HSC) niche contain precursors that reform the niche. CB ossicles were regularly observed upon transplantation. Compared with BM ossicles, CB ossicles showed a predominance of red marrow over yellow marrow, as demonstrated by histomorphological analyses and the number of hematopoietic cells isolated within ossicles. Marrow cavities from CB and BM ossicles included donor-derived CD146-expressing osteoprogenitors and host-derived mature hematopoietic cells, clonogenic lineage-committed progenitors and HSCs. Furthermore, human CD34+ cells transplanted into ossicle-bearing mice engrafted and maintained human HSCs in the niche. Our data indicate that CB-BFs are able to recapitulate the conditions by which the bone marrow microenvironment is formed and establish complete HSC niches, which are functionally supportive of hematopoietic tissue.


PTMOracle: A Cytoscape App for Covisualizing and Coanalyzing Post-Translational Modifications in Protein Interaction Networks.
Post-translational modifications of proteins (PTMs) act as key regulators of protein activity and of protein-protein interactions (PPIs). To date, it has been difficult to comprehensively explore functional links between PTMs and PPIs. To address this, we developed PTMOracle, a Cytoscape app for coanalyzing PTMs within PPI networks. PTMOracle also allows extensive data to be integrated and coanalyzed with PPI networks, allowing the role of domains, motifs, and disordered regions to be considered. For proteins of interest, or a whole proteome, PTMOracle can generate network visualizations to reveal complex PTM-associated relationships. This is assisted by OraclePainter for coloring proteins by modifications, OracleTools for network analytics, and OracleResults for exploring tabulated findings. To illustrate the use of PTMOracle, we investigate PTM-associated relationships and their role in PPIs in four case studies. In the yeast interactome and its rich set of PTMs, we construct and explore histone-associated and domain-domain interaction networks and show how integrative approaches can predict kinases involved in phosphodegrons. In the human interactome, a phosphotyrosine-associated network is analyzed but highlights the sparse nature of human PPI networks and lack of PTM-associated data. PTMOracle is open source and available at the Cytoscape app store: http://apps.cytoscape.org/apps/ptmoracle .


A mixed method approach to evaluating eating-related psychopathologies in collegiate student-athletes.
To examine the presence of body image concerns, drive for muscularity, and disordered eating behaviors in collegiate student-athletes. One hundred and one NCAA Division I student-athletes participated in Phase I; 15 of these also participated in Phase II. This study employed a mixed method, sequential explanatory design. Participants first completed survey measures assessing body image concern, drive for muscularity, and eating behaviors. These results influenced open-ended, semi-structured interviews, which were thematically analyzed. Body image and disordered eating behaviors were of greater concern than drive for muscularity. Student-athletes reported engaging in eating behaviors as opposed to not eating, yet these eating behaviors trended toward disordered behaviors such as obsessive "healthy eating" or orthorexia. This study took a novel methodological approach to examining student-athlete body image and eating behaviors. Results emphasize the need for further support and education for student-athletes around body image and eating behaviors.


Differences in the ultimate strength of the biceps anchor and the generation of type II superior labral anterior posterior lesions in a cadaveric model.
The pathogenesis of superior labral anterior posterior lesions remains controversial. The biceps anchor is more vulnerable to loading with a posterior vector as opposed to an in-line pull. Controlled laboratory study. Eight pairs of cadaveric shoulders were dissected, and the biceps tendon was loaded to failure in 1 of 2 loading patterns. Loading pattern A was meant to simulate the eccentric load of the biceps in the deceleration phase of throwing; loading pattern B was meant to simulate the posterior biceps load of the late cocking phase. The biceps anchor demonstrated significantly increased ultimate strength with in-line loading (group A, 508 N) as opposed to posterior loading (group B, 262 N, P < .001). All group B specimens failed at the biceps anchor, resulting in a type II superior labral anterior posterior lesion. Specimens in group A did not create a superior labral anterior posterior lesion. Direction of biceps loading resulted in significant differences in the ultimate strength of the biceps anchor and the generation of superior labral anterior posterior lesions. The biceps anchor was significantly weaker when loaded with a posterior vector. The superior labrum may be most vulnerable to injury in late cocking. The reproducible generation of type II superior labral anterior posterior lesions may have applications as a biomechanical model.