Synergy between sulforaphane and selenium in the up-regulation of thioredoxin reductase and protection against hydrogen peroxide-induced cell death in human hepatocytes.
Dietary isothiocyanates and selenium are chemopreventive agents and potent inducers of antioxidant enzymes. It has been previously shown that sulforaphane and selenium have a synergistic effect on the upregulation of thioredoxin reductase-1 (TrxR-1) in human hepatoma HepG2 cells. In this paper, further evidence is presented to show that sulforaphane and selenium synergistically induce TrxR-1 expression in immortalised human hepatocytes. Sulforaphane was found to be more toxic toward hepatocytes than HepG2 cells with IC50=25.1 and 56.4 μM, respectively. Sulforaphane can protect against hydrogen peroxide-induced cell death and this protection was enhanced by co-treatment with selenium. Using siRNA to knock down TrxR-1 or Nrf2, sulforaphane (5 μM)-protected cell viability was reduced from 73% to 46% and 34%, respectively, suggesting that TrxR-1 is an important enzyme in protection against hydrogen peroxide-induced cell death. Sulforaphane-induced TrxR-1 expression was positively associated with significant levels of Nrf2 translocation into the nucleus, but co-treatment with selenium showed no significant increase in Nrf2 translocation. Moreover, MAPK (ERK, JNK and p38) and PI3K/Akt signalling pathways were found to play no significant role in sulforaphane-induced Nrf2 translocation into the nucleus. However, blocking ERK and JNK signalling pathways decreased sulforaphane-induced TrxR-1 mRNA by about 20%; whereas blocking p38 and PI3K/AKT increased TrxR-1 transcription. In summary, a combination of sulforaphane and selenium resulted in a synergistic upregulation of TrxR-1 that contributed to the enhanced protection against free radical-mediated oxidative damage in human hepatocytes.


Error-resilient image and video transmission over the Internet using unequal error protection.
This paper presents a new bit-plane-wise unequal error protection algorithm for progressive bitstreams transmitted over lossy networks. The proposed algorithm protects a compressed embedded bitstream generated by a 3-D SPIHT algorithm by assigning an unequal amount of forward error correction (FEC) to each bit-plane. The proposed algorithm reduces the amount of side information needed to send the size of each code to the decoder by limiting the number of quality levels to the number of bit-planes to be sent while providing a graceful degradation of picture quality as packet losses increase. We also apply our proposed algorithm to transmission of JPEG 2000 coded images over the Internet. To get additional error-resilience at high packet loss rates, we extend our algorithm to multiple-substream unequal error protection. Simulation results show that the proposed algorithm is simple, fast and robust in hostile network conditions and, therefore, can provide reasonable picture quality for video applications under varying network conditions.


DNA copy number aberrations associated with aneuploidy and chromosomal instability in breast cancers.
Biological characteristics of a tumor are primarily affected by its genomic alterations. It is thus important to ascertain whether there are genomic changes linked with DNA ploidy and/or chromosomal instability (CIN). In the present study, using fresh-frozen samples of 46 invasive breast cancers, laser scanning cytometry, array-based comparative genomic hybridization, and chromosome fluorescence in situ hybridization were performed to assess DNA ploidy, DNA copy number aberrations (DCNAs), and CIN status. Both ploidy and CIN status were examined in 36 tumors, resulting in 23 aneuploid/CIN+ tumors, 1 aneuploid/CIN-, 2 diploid/CIN+, and 10 diploid/CIN- tumors. Comparison of the aCGH data with the DNA ploidy and CIN status identified cytogenetically 11 characteristic breast cancers with distinctive DCNAs. The 11 tumors were classified into two types; one type is diploid/CIN- phenotype containing 4 DCNAs, and the other aneuploid/CIN+ phenotype containing 7 DCNAs. In 30 (65.2%) of the 36 breast cancers, the status of DNA ploidy and CIN depended on the type of DCNAs. Furthermore, the DNA ploidy phenotype depended on the dominant type of DCNAs even in tumors with a mixture of multiple DCNAs of one type and a single DCNA of the other type. Tumors with multiple DCNAs of both types represented aneuploidy and over three quarters of breast cancers carry at least one type of the DCNAs. These results suggested that, in breast cancers, the status of DNA ploidy and CIN was likely to determine at the beginning of carcinogenesis.


One year update on the COVID-19 pandemic: Where are we now?
We are living through an unprecedented crisis with the rapid spread of the new coronavirus disease (COVID-19) worldwide within a short time. The timely availability of thousands of SARS-CoV-2 genomes has enabled the scientific community to study the origin, structures, and pathogenesis of the virus. The pandemic has spurred research publication and resulted in an unprecedented number of therapeutic proposals. Because the development of new drugs is time consuming, several strategies, including drug repurposing and repositioning, are being tested to treat patients with COVID-19. Researchers have developed several potential vaccine candidates that have shown promise in phase II and III trials. As of 12 November 2020, 164 candidate vaccines are in preclinical evaluation, and 48 vaccines are in clinical evaluation, of which four have cleared phase III trials (Pfizer/BioNTech's BNT162b2, Moderna's mRNA-1273, University of Oxford & AstraZeneca's AZD1222, and Gamaleya's Sputnik V vaccine). Despite the acquisition of a vast body of scientific information, treatment depends only on the clinical management of the disease through supportive care. At the pandemic's 1-year mark, we summarize current information on SARS-CoV-2 origin and biology, and advances in the development of therapeutics. The updated information presented here provides a comprehensive report on the scientific progress made in the past year in understanding of SARS-CoV-2 biology and therapeutics.


An evaluation of the role of risk-based decision-making in a former manufactured gas plant site remediation.
Environmental remediation decisions are driven by the need to minimize human health and ecological risks posed by environmental releases. The Risk Assessment Guidance for Superfund Sites enunciates the principles of exposure and risk assessment that are to be used for reaching remediation decisions for sites under Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). Experience with remediation management under CERCLA has led to recognition of some crucial infirmities in the processes for managing remediation: cleanup management policies are ad hoc in character, mandates and practices are strongly conservative, and contaminant risk management occurs in an artificially narrow context. The purpose of this case study is to show how a policy of risk-based decision-making was used to avoid customary pitfalls in site remediation. This case study describes the risk-based decision-making process in a remedial action program at a former manufactured gas plant site that successfully achieved timely and effective cleanup. The remediation process operated outside the confines of the CERCLA process under an administrative consent order between the utility and the New Jersey Department of Environmental Protection. A residential use end state was negotiated as part of this agreement. The attendant uncertainties, complications, and unexpected contingencies were overcome by using the likely exposures associated with the desired end state to structure all of the remediation management decisions and by collecting site-specific information from the very outset to obtain a detailed and realistic characterization of human health risks that needed to be mitigated. The lessons from this case study are generalizable to more complicated remediation cases, when supported by correspondingly sophisticated technical approaches.


Thermally oxidized dietary fats increase the susceptibility of rat LDL to lipid peroxidation but not their uptake by macrophages.
The aim of this study was to investigate the effect of dietary oxidized fats on the lipoprotein profile and the atherogenicity of LDL. Two experiments with male Sprague-Dawley rats were conducted. In Experiment 1, diets with either fresh fat or oxidized fat, prepared by heating at temperatures of 50, 105 or 190 degrees C, containing either 25 or 250 mg alpha-tocopherol equivalents/kg were used. In Experiment 2, diets with fresh or oxidized fat, heated at a temperature of 55 degrees C, containing 25 mg alpha-tocopherol equivalents/kg, were used. In Experiment 1, rats fed all types of oxidized fats had higher concentrations of HDL cholesterol and lower ratios between plasma and HDL cholesterol than rats fed the diet containing the fresh fat. As determined from the lag time, the susceptibility of LDL to copper-induced lipid peroxidation was higher in rats fed oxidized fats heated at 105 or 190 degrees C than in rats fed the diets containing the fresh fat or the oxidized fat treated at 50 degrees C, irrespective of the dietary vitamin E concentration. However, in Experiment 2, the composition of LDL apolipoproteins and uptake of LDL by macrophages were not different between rats fed the fresh fat diet and those fed the oxidized fat diet. We conclude that ingestion of oxidized fats does not adversely affect the lipoprotein profile in the rat model used, and does not cause modifications of apolipoproteins that would lead to enhanced uptake of LDL via macrophage scavenger receptors.


Josephson tunnel-junction electrode materials.
Josephson superconducting devices of the tunnel-junction type have exciting potential for use in building ultrahigh-speed computers. We consider the properties of superconducting metals that are needed for such devices that would be used in integrated computer circuits operated at a temperature near absolute zero. Recent advances in lead-alloy thin-film materials are described that have led to substantial improvements in lead-alloy Josephson device reliability. The properties of a Pb(0.84)In(0.12)Au(0.04) alloy, of Nb, and of Nb(3)Sn are discussed as examples of three different groups of materials that are of interest. Investigations of lead-alloy and niobium devices have progressed to the point that it is evident that they have good potential for fabricating integrated circuits containing large numbers of devices.


Regulatory role of cAMP on expression of Cdk4 and p27(Kip1) by inhibiting phosphatidylinositol 3-kinase in corneal endothelial cells.
Fibroblast growth factor (FGF)-2 is a potent mitogen of corneal endothelial cells (CECs). Results in an earlier study showed that FGF-2 activates phosphatidylinositol (PI) 3-kinase to stimulate the cell cycle machinery. The current study was designed to determine whether adenosine 3',5'-monophosphate (cAMP) antagonizes FGF-2 by inhibiting PI 3-kinase/Akt pathways, thus leading to regulation of cyclin-dependent kinase 4 (Cdk4) and p27(Kip1) (p27) expression. Cell proliferation was assayed by counting cells. Subcellular localization of proteins was determined by immunofluorescent staining and expression of Cdk4, p27, PI 3-kinase, Akt, and beta-actin was analyzed by immunoblot analysis. PI 3-kinase activity was determined by measuring production of phosphatidylinositol-3-phosphate. 8-Bromoadenosine cAMP (8-Br-cAMP), a diffusible cAMP analogue, inhibited the PI 3-kinase/Akt signaling pathways. The 8-Br-cAMP and PI 3-kinase inhibitor (LY294002) produced equivalent stimulation and inhibition, respectively, of p27 and Cdk4 protein levels. They also equally inhibited cell proliferation, nuclear translocation of Cdk4, and phosphorylation of p27. Negative regulation of PI 3-kinase by 8-Br-cAMP was mediated by a direct inhibition of PI 3-kinase activity, which subsequently blocked phosphorylation of Akt at both the Ser473 and Thr308 sites. In addition, 8-Br-cAMP promoted a rapid turnover of Akt protein, and 8-Br-cAMP markedly reduced the half-life of Cdk4 protein. This inhibitory activity of cAMP was not mediated by PKA, but 8-Br-cAMP inhibited membrane localization of the p85 regulatory subunit of PI 3-kinase. These data support the hypothesis that cAMP inhibits the proliferation of CECs, preventing them from entering the S phase by negatively regulating PI 3-kinase.


A Preliminary Experimental Investigation of Emotion Dysregulation and Impulsivity in Risky Behaviours.
This prospective experimental study sought to examine the unique effects of emotion dysregulation and impulsivity on risky behaviors over time. To this end, 20 African American women enrolled in a historically black university in the southern United States were randomly assigned to receive one of two brief empirically-supported skills training modules (i.e., emotion modulation [EM] or impulsivity reduction [IR]). Participants completed measures of emotion dysregulation, impulsivity, and past-week risky behaviors before (pre-) and one week after (post-) the experimental manipulation. Participants assigned to the EM condition reported significant improvements from pre- to post-manipulation in overall emotion dysregulation (as well as all specific dimensions of emotion dysregulation other than lack of emotional awareness), as well as two dimensions of impulsivity: negative and positive urgency. Participants assigned to the IR condition reported a significant decrease in one dimension of impulsivity (lack of premeditation) from pre- to post-manipulation. Findings also revealed a significant effect of time on risky behaviors, with participants reporting significantly fewer past-week risky behaviors at the post- (vs. pre-) manipulation assessment. Finally, changes in emotion dysregulation from pre- to post-manipulation accounted for the observed reduction in risky behaviors over time (above and beyond changes in impulsivity dimensions). Results highlight the relevance of emotion dysregulation to risky behaviors and suggest that treatments targeting emotion dysregulation may be useful in reducing risky behaviors.


Aerobic biodegradation of 2,4-DNT and 2,6-DNT: performance characteristics and biofilm composition changes in continuous packed-bed bioreactors.
This manuscript deals with continuous experiments for biodegradation of individual dinitrotoluenes by a defined mixed culture in packed-bed reactors (PBRs) containing either poraver or fire-clay as packing material. Removal efficiencies and volumetric biodegradation rates were measured as a function of the loading rate of 2,4-dinitrotoluene (2,4-DNT) and 2,6-dinitrotoluene (2,6-DNT) under steady-state conditions. The poraver reactor showed higher removal efficiencies for both the DNTs. The removal efficiency for 2,4-DNT remained greater than 90% in the poraver reactor whereas it dropped steadily from 85 to 65% in the fire-clay reactor as the organic loading rates were increased from 19 to 60 mg L(-1)day(-1). Similar trends were seen for the volumetric degradation rate as well. In both the reactors, 2,4-DNT degraded more effectively than 2,6-DNT. The microbial consortium was characterized both in the inoculum as well as in the operating PBR. Cell numbers per gram dry packing material were similar in the two reactors. However, there was a distinct difference in the nature of microorganisms that were found in the two packings. The fire-clay contained a larger number of cells that were not primary degraders of DNTs.


Comparing the Palliative Care Needs of Those With Cancer to Those With Common Non-Cancer Serious Illness.
Historically, palliative care has been focused on those with cancer. Although these ties persist, palliative care is rapidly integrating into the care of patients with common, non-cancer serious illnesses. Despite this, the bulk of literature informing palliative care practices stems from the care of cancer patients. We compared functionality, advanced care planning, hospital admissions, prognosis, quality of life, pain, dyspnea, fatigue, and depression between patients with cancer and three non-cancer diagnoses-end-stage renal disease (ESRD), heart failure (HF), and chronic obstructive pulmonary disease (COPD). We conducted a cross-sectional, retrospective analysis of the characteristics and symptoms of patient's with ESRD, HF, COPD, and cancer at time of first specialty palliative care referral. Using a web-based point of care quality assessment and reporting tool, Quality Data and Collection Tool-Palliative care, this analysis evaluated all eligible patients who received a palliative care consultation between October 1, 2012 and November 25, 2014. Data were obtained from 13 participating sites. The primary outcome for the study was functionality using the palliative performance scale. Hospital admission in the last 30 days, prognosis, patient's understanding of prognosis, advanced care planning including code status and appointed decision maker, pain, fatigue, depression, and dyspnea were also evaluated as secondary outcomes. We tested for an association between our outcomes with disease type (cancer vs. non-cancer) fitting multivariable logistic regression models. We found that the patients with primary diagnoses other than cancer were less functional at time of referral (odds ratio: 1.6; 95% CI: 1.1, 2.3; P < 0.05). Patients with COPD, ESRD, and HF were less functional and more likely to be hospitalized at time of referral to palliative care than cancer patients. These findings may be reflective of the slower and more varied trajectory of non-cancer serious illness. One aim of palliative care for those with non-cancer severe illness should be directed toward improving and assisting with functionality and decreasing frequency of hospital admissions. These interventions could take place in the palliative care office, but could also be integrated into hospital discharge plans.


Storage of conidia of Penicillium chrysogenum in liquid nitrogen.
Conidiated slope cultures of derivative of Penicillium chrysogenum Wis 54-1255 were stored at -196 or +4 C for a period of 3.5 years. After this time, the viability fell to 68% in the former case and to 4% in the latter. At the end of the experiment, 65 single conidial isolates from each series were tested for penicillin yield. Among those from conidia stored at -196 C, the spread of penicillin yields did not differ markedly from that of 65 single conidial isolates made as controls prior to storage. However, 18% of those from conidia stored at +4 C formed a subpopulation with substantially lower penicillin titers than those of control isolates. Storage at -196 C may reduce or prevent a possible source of penicillin yield decay, namely, the selection of spontaneous mutants of low titer present in small numbers in the original culture and selected, as viability decreased, by virtue of their increased longevity relative to that of the parental culture.


Pattern of rash, diarrhea, and hepatic toxicities secondary to lapatinib and their association with age and response to neoadjuvant therapy: analysis from the NeoALTTO trial.
We investigated the pattern of rash, diarrhea, and hepatic adverse events (AEs) secondary to lapatinib and their association with age and pathologic complete response (pCR) in the Neoadjuvant Lapatinib and/or Trastuzumab Treatment Optimisation (NeoALLTO) phase III trial. Patients with HER2-positive early breast cancer were randomly assigned to receive lapatinib (Arm A), trastuzumab (Arm B), or their combination (Arm C) for 6 weeks followed by the addition of paclitaxel for 12 weeks before surgery. We investigated the frequency and time to developing each AE according to age (≤ 50 v > 50 years) and their association with pCR in a logistic regression model adjusted for age, hormone receptors, tumor size, nodal status, planned breast surgery, completion of lapatinib administration, and treatment arm. Only patients randomly assigned to arms A and C were eligible (n = 306). Younger patients (≤ 50 years) experienced significantly more rash compared with older patients (74.4% v 47.9%; P < .0001). Diarrhea and hepatic AEs were observed in 78.8% and 41.2% of patients, respectively, with no differences in rate or severity or time of onset according to age. Early rash (ie, before starting paclitaxel) was independently associated with a higher chance of pCR, mainly in patients older than 50 years (odds ratio [OR] = 3.76; 95% CI, 1.69 to 8.34) but not in those ≤ 50 years (OR = 0.92; 95% CI, 0.45 to 1.88; P for interaction = .01). No significant association was observed between pCR and diarrhea or hepatic AEs. Our results indicate that the frequency and clinical relevance of lapatinib-related rash is largely dependent on patient age.


The induction of mutants of Acinetobacter calcoaceticus NCIB8250 and their selection by vancomycin.
The mutagenic and lethal effects of N-methyl-N'-nitro-N-nitrosoguanidine (NTG), ethyl methanesulphonate (EMS), ultraviolet light irradiation and near-ultraviolet light irradiation with 8-methoxypsoralen on the bacterium Acinetobacter calcoaceticus NCIB8250 were examined. The production of auxotrophic mutants was used as a measure of mutagenic efficiency. Under appropriate conditions all four agents were mutagenic. EMS and NTG although more effective than irradiation, did not cause such a high frequency of mutation as has been observed with other bacteria. A combination of vancomycin and penicillin V gave enrichment of non-metabolizing bacteria and optimum conditions were found for the use of these compounds in a selection technique.


Predictors of early remission of hyperthyroidism in children.
Children with hyperthyroidism often require prolonged courses of antithyroid medication to achieve remission, and long-term compliance is problematic. To determine which clinical and laboratory features predict early remission, we reviewed the records of 191 patients less than 19 yr old with Graves' disease. We compared patients achieving remission within 2 yr (group 1, n = 27) with those who completed more than 2 yr of medical therapy but did not achieve a remission (group 2, n = 79). Patients who were in neither of the above categories (n = 85) were excluded from the statistical analysis. Variables that were measurable at the time of diagnosis, recorded in more than 50% of the study population and associated with early remission in the univariate analysis (P < or = 0.05), were entered into a stepwise multiple logistic regression analysis. Variables retaining a significant association with early remission (P < 0.05) were considered independent predictors of early remission. Patients achieving early remission were older (mean, 12.5 vs. 10.9 yr, P = 0.039) and had higher body mass indexes (BMI, 19.0 vs. 16.6, P = 0.002), higher BMI SD scores (-0.03 vs. -0.60, P = 0.004), lower heart rates (110 vs. 121, P = 0.023), smaller goiters (group 1: 60% with moderate/large goiter; group 2: 83%, P = 0.050), lower platelet counts (272 vs. 339 K/microL, P = 0.006), lower serum T4 and T3 concentrations at presentation (T4: 18.3 vs. 22.5 microg/dL, P = 0.015; T3: 439 vs. 613 ng/dL, P = 0.008), and were less likely to have a positive test for thyroid stimulating Igs (group 1: 50% vs. group 2: 93%, P = 0.008). Regression analysis identified BMI SD score and goiter size as independent predictors of early remission (P < 0.05). Eighty-six percent of patients with BMI SD score above -0.5 SD and minimal/small goiters achieved early remission, compared with 13% of those with BMI SD score below -0.5 SD and moderate/large goiters. We conclude that, of multiple clinical and laboratory variables associated with early remission, BMI SD score and goiter size are independent predictors. Algorithms employing these two variables can be used to facilitate counseling of patients and expedite therapeutic decisions.


Incidence, prevalence and mortality of patients with psoriasis: a U.K. population-based cohort study.
The burden of psoriasis across many world regions is high and there is a recognized need to better understand the epidemiology of this common skin disorder. To examine changes in the prevalence and incidence of psoriasis, and mortality rates over a 15-year period. Cohort study involving analysis of longitudinal electronic health records between 1999 and 2013 using the U.K. Clinical Practice Research Datalink (CPRD). The prevalence of psoriasis increased steadily from 2·3% (2297 cases per 100 000) in 1999 to 2·8% (2815 per 100 000) in 2013, which does not appear to be attributable to changes in incidence rates. We observed peaks in age bands characteristic of early-onset (type I) and late-onset (type II) psoriasis, and changes in incidence and prevalence rates with increasing latitude in the U.K. All-cause mortality rates for the general population and for patients with psoriasis have decreased over the last 15 years. However, the risk of all-cause mortality for patients with psoriasis remains elevated compared with people without psoriasis (hazard ratio 1·21; 95% confidence interval 1·13-1·3) and we found no significant change in this relative excess mortality gap over time. We found an increasing population living longer with psoriasis in the U.K., which has important implications for healthcare service delivery and for resource allocation. Importantly, early mortality in patients with psoriasis remains elevated compared with the general population and we found no evidence of change in this premature mortality gap.


Vascular endothelial growth factor blockade promotes the transition from compensatory cardiac hypertrophy to failure in response to pressure overload.
Cardiac hypertrophy is associated with upregulation of vascular endothelial growth factor (VEGF) in the myocardium. Here, we evaluated the effects of a decoy VEGF receptor on heart morphology and function to a murine model of pressure overload hypertrophy. Mice were administered adenoviral vector encoding a decoy VEGF receptor (Ad-Flk), and their hearts were subjected to pressure overload by transverse aortic constriction (TAC). Treatment with Ad-Flk led to a net reduction in capillary density in hearts subjected to TAC. Ad-Flk also led to a reduction in TAC-induced cardiac hypertrophy and promoted left ventricle dilatation and a loss in contractile function. Treatment with Ad-Flk markedly increased myocardial fibrosis and collagen gene upregulation. In contrast, Ad-Flk had no effect on any of these parameters in sham-treated mice. Administration of a VEGF trap reagent diminished pressure overload cardiac hypertrophy and promoted the progression to heart failure but had no effect on sham-treated animals. These findings suggest that VEGF is required to maintain myocardial capillary density and that reductions in the vascular bed are associated with the transition from compensatory hypertrophy to failure.


First principles investigation of the spectral properties of neutral, zwitterionic, and bis-cationic azaacenes.
An in-depth investigation of the optical properties of recently-synthesized linear azaacene derivatives of various electronic nature (neutral, dicationic, and zwitterionic) is presented. Our simulations include not only the determination of the vertical transitions, but also of the adiabatic energies and vibrationally-resolved spectra using the adiabatic Hessian approach in connection with the Time-Dependent Density Functional Theory (TD-DFT) and second-order Coupled-Cluster (CC2) methods. We show that the theoretical results are in excellent agreement with experiment for both the λmax and the band topologies, a statement holding in the full visible domain. We also analyse the nature of the key vibrations explaining the specific band shapes of these compounds. In addition, we investigate a series of yet-unknown substituted systems in an effort to design new compounds with improved (redshifted) properties for optoelectronics applications.


Molecular cloning, characterization, and expression of the M antigen of Histoplasma capsulatum.
The major diagnostic antigens of Histoplasma capsulatum are the H and M antigens, pluripotent glycoproteins that elicit both humoral and T-cell-mediated immune responses. These antigens may play a role in the pathogenesis of histoplasmosis. M antigen is considered immunodominant because antibodies against it are the first precipitins to arise in acute histoplasmosis and are commonly present during all phases of infection. The biological activity of monomolecular M antigen and its ability to elicit a protective immune response to H. capsulatum are largely unknown. A molecular approach was used to identify the biological nature of M antigen, including its purification from histoplasmin, partial digestion with proteinases, and reverse-phase high-performance liquid chromatography to separate the released peptides. The amino acid sequences of the purified peptides were obtained by Edman degradation, and using degenerate oligonucleotide primers for PCR, a 321-bp fragment of the gene encoding the M antigen was amplified from genomic H. capsulatum DNA. This fragment was used to screen an H. capsulatum genomic DNA library, leading to the isolation, cloning, and sequencing of the full-length gene. The M gene consists of 2, 187-bp DNA encoding a protein of 80,719 Da, which has significant homology to catalases from Aspergillus fumigatus, Aspergillus niger, and Eimericella nidulans. A cDNA was generated by reverse transcription-PCR and cloned into the expression vector pQE40. The identity of the cloned, expressed protein was confirmed by Western blotting. The recombinant fusion protein was immunoreactive with monoclonal antibodies raised against M antigen, with polyclonal mouse anti-M antiserum, and with a serum sample from a patient with histoplasmosis. The gene encoding the major immunodominant M antigen of H. capsulatum is a presumptive catalase, and the recombinant protein retains serodiagnostic activity.


Cholesterol-rich synovial effusions.
The occurrence of cholesterol-rich synovial fluids in 2 patients with rheumatoid arthritis is described. These fluids are rare, and are readily mistaken for pus. Their origins are discussed in relation to chronic inflammation.


The electric field induced in the brain by magnetic stimulation: a 3-D finite-element analysis of the effect of tissue heterogeneity and anisotropy.
We investigate the effect of tissue heterogeneity and anisotropy on the electric field and current density distribution induced in the brain during magnetic stimulation. Validation of the finite-element (FE) calculations in a homogeneous isotropic sphere showed that the magnitude of the total electric field can be calculated to within an error of approximately 5% in the region of interest, even in the presence of a significant surface charge contribution. We used a high conductivity inclusion within a sphere of lower conductivity to simulate a lesion due to an infarct. Its effect is to increase the electric field induced in the surrounding low conductivity region. This boost is greatest in the vicinity of interfaces that lie perpendicular to the current flow. For physiological values of the conductivity distribution, it can reach a factor of 1.6 and extend many millimeters from the interface. We also show that anisotropy can significantly alter the electric field and current density distributions. Either heterogeneity or anisotropy can introduce a radial electric field component, not present in a homogeneous isotropic conductor. Heterogeneity and anisotropy are predicted to significantly affect the distribution of the electric field induced in the brain. It is, therefore, expected that anatomically faithful FE models of individual brains which incorporate conductivity tensor data derived from diffusion tensor measurements, will provide a better understanding of the location of possible stimulation sites in the brain.


[Antibodies against p-aminophenyl-beta-D-galactopyranoside-containing proteins].
The antibodies were prepared from antisera of rabbits immunized with bovine serum albumin containing covalently bound p-aminophenyl-beta-D-galactopyranoside (APG) and purified by affinity chromatography on APG-containing ovalbumin immobilized by BrCN-activated Sepharose 4B. The antibodies possessed a selective specificity for APG and interacted with different APG-containing proteins, including APG-containing lysosomal alpha-glucosidase. The purified antibodies are immunoglobulins of G type as was determined from the molecular weights of native and dissociated antibodies and from the immunochemical assays with antibodies against rabbit IgG.


Reliability Fusion of Time-of-Flight Depth and Stereo Geometry for High Quality Depth Maps.
Time-of-flight range sensors have error characteristics, which are complementary to passive stereo. They provide real-time depth estimates in conditions where passive stereo does not work well, such as on white walls. In contrast, these sensors are noisy and often perform poorly on the textured scenes where stereo excels. We explore their complementary characteristics and introduce a method for combining the results from both methods that achieve better accuracy than either alone. In our fusion framework, the depth probability distribution functions from each of these sensor modalities are formulated and optimized. Robust and adaptive fusion is built on a pixel-wise reliability weighting function calculated for each method. In addition, since time-of-flight devices have primarily been used as individual sensors, they are typically poorly calibrated. We introduce a method that substantially improves upon the manufacturer's calibration. We demonstrate that our proposed techniques lead to improved accuracy and robustness on an extensive set of experimental results.


Striking variation in coronary heart disease mortality in the United States among black and white women aged 45-54 by state.
Coronary heart disease (CHD) is the leading cause of death for women in the United States. There has been a substantial decrease in CHD mortality in the past few decades in the United States for both women and men. The change in lifestyle after World War II may affect the incidence of and mortality from CHD in a more recent birth cohort, such as the 45-54-year-old age group. CHD mortality among women aged 45-54 by state and race in the United States and other countries in 1994 was evaluated. Correlation of the CHD mortality with state-specific and race-specific educational attainment (% of not a high school graduate) and state-specific prevalence of smoking was examined. There was a 2.5-fold difference in CHD mortality (ICD 410-414, 429.2) between black and white women aged 45-54: 78/100,000 for black versus 31/100,000 for white women; a 3-fold difference in CHD mortality among white women by state: 16/100,000 in Colorado versus 53/100,000 in Louisiana; and a 3-fold difference in CHD mortality among black women by state: 45/100,000 in New Jersey versus 124/100,000 in Arkansas. CHD mortality was correlated with educational attainment among white women (r = 0.62, p = 0.001) and with prevalence of smoking (r = 0.39, p = 0.021). There is a large variation in CHD mortality among women aged 45-54 in the United States by race and state. These differences may reflect variations in coronary risk factors. More detailed evaluation of determinants of CHD mortality by area is needed, as are public health programs that can reduce the marked disparity in CHD mortality in the United States.


Standardizing postoperative care for pediatric intradural Chiari decompressions to decrease length of stay.
Amid national and local budget crises, cutting costs while maintaining quality care is a top priority. Chiari malformation is a relatively common pediatric neurosurgical pathology, and postoperative care varies widely. The postoperative course can be complicated by pain and nausea, which can extend the hospital stay. In this study, the authors aimed to examine whether instituting a standardized postoperative care protocol would decrease overall patient hospital length of stay (LOS) as well as cost to families and the hospital system. A retrospective study of pediatric patients who underwent an intradural Chiari decompression with expansile duraplasty at a single institution from January 2016 to September 2019 was performed. A standardized postoperative care protocol was instituted on May 17, 2018. Pre- and postprotocol groups were primarily analyzed for demographics, LOS, and the estimated financial expense of the hospital stay. Secondary analysis included readmissions, opioid consumption, and follow-up. The analysis included 132 pediatric patients who underwent an intradural Chiari decompression with expansile duraplasty. The preprotocol group included 97 patients and the postprotocol group included 35 patients. Patient age ranged from 0.5 to 26 years (mean 9.5 years). The mean LOS preprotocol was 55.48 hours (range 25.90-127.77 hours), and the mean postprotocol LOS was 46.39 hours (range 27.58-77.38 hours). The comparison between means showed a statistically significant decrease following protocol initiation (95% CI 1.87-16.31 hours, p = 0.014). In the preprotocol group, 21 of 97 patients (22%) were discharged the first day after surgery compared with 14 of 35 patients (40%) in the postprotocol group (p = 0.045). The estimated cost of one night on the pediatric neurosurgical intermediate ward was approximately $4500, which gives overall cost estimates for 100 theoretical cases of $927,800 for the preprotocol group and $732,900 for the postprotocol group. By instituting a Chiari protocol, postoperative LOS was significantly decreased, which resulted in decreased healthcare costs while maintaining high-quality and safe care.


Spatiotemporal Crossover between Low- and High-Temperature Dynamical Regimes in the Quantum Heisenberg Magnet.
The stranglehold of low temperatures on fascinating quantum phenomena in one-dimensional quantum magnets has been challenged recently by the discovery of anomalous spin transport at high temperatures. Whereas both regimes have been investigated separately, no study has attempted to reconcile them. For instance, the paradigmatic quantum Heisenberg spin-1/2 chain falls at low temperature within the Tomonaga-Luttinger liquid framework, while its high-temperature dynamics is superdiffusive and relates to the Kardar-Parisi-Zhang universality class in 1+1 dimensions. This Letter aims at reconciling the two regimes. Building on large-scale matrix product state simulations, we find that they are connected by a temperature-dependent spatiotemporal crossover. As the temperature T is reduced, we show that the onset of superdiffusion takes place at longer length and timescales ∝1/T. This prediction has direct consequences for experiments including nuclear magnetic resonance: it is consistent with earlier measurements on the nearly ideal Heisenberg S=1/2 chain compound Sr_{2}CuO_{3}, yet calls for new and dedicated experiments.


Hepatitis B virus DNA splicing in Lebanese blood donors and genotype A to E strains: implications for hepatitis B virus DNA quantification and infectivity.
Hepatitis B virus (HBV) is one of the major viruses transmissible by blood that causes chronic infection in immunocompromised individuals. The study of 61 HBV carrier blood donors from Lebanon revealed multiple patterns of spliced HBV DNA. HBV DNA splicing was examined and quantified in samples of five genotypes and in seroconversion panels. The Lebanese sample median viral load was 1.5 ×10(2) IU/ml. All strains were genotype D, serotype ayw; 35 clustered as subgenotype D1 and 7 clustered as subgenotype D2. Three splice variants (SP1, SP1A, and Pol/S) were observed in 12 high-viral-load samples. Twenty samples of each genotype, A to E, were tested for the presence of HBV spliced DNA and SP1-specific splice variant. An unspliced HBV genome was dominant, but 100% of strains with a viral load of ≥10(5) copies/ml contained various proportions of spliced DNA. SP1 was detected in 56/100 (56%) samples in levels that correlated with the overall viral load. HBV DNA quantification with S (unspliced) and X (total DNA) regions provided different levels of viral load, with the difference corresponding to spliced DNA. During the highly infectious window period, the SP1 variant became detectable shortly after the hepatitis B surface antigen (HBsAg), suggesting a correlation between the initiation of splicing and the production of detectable levels of HBsAg. The quantification of HBV DNA with primers located outside and inside the spliced region might provide different estimations of viral load and differentiate between infectious and defective viral genomes. The role of splicing neoproteins in HBV replication and interaction with the host remains to be determined.


[Diseases of the lumbosacral division of the peripheral nervous system in children and adolescents].
The clinical picture and course of diseases affecting the lumbosacral division of the peripheral nervous system in 111 children and adolescents aged 8 to 18 years are described. It was found that in 40.5% of the patients the diseases developed in the presence of congenital anomalies in the development of the lumbosacral part of the vertebral column. Unlike adults, the pain syndrome in children and adolescents is less pronounced, and leading in the clinical picture are signs of reflectory-tonic protection of the vertebral column.


Last rolls of the yoyo: Assessing the human canonical protein count.
In 2004, when the protein estimate from the finished human genome was only 24,000, the surprise was compounded as reviewed estimates fell to 19,000 by 2014. However, variability in the total canonical protein counts (i.e. excluding alternative splice forms) of open reading frames (ORFs) in different annotation portals persists. This work assesses these differences and possible causes. A 16-year analysis of Ensembl and UniProtKB/Swiss-Prot shows convergence to a protein number of ~20,000. The former had shown some yo-yoing, but both have now plateaued. Nine major annotation portals, reviewed at the beginning of 2017, gave a spread of counts from 21,819 down to 18,891. The 4-way cross-reference concordance (within UniProt) between Ensembl, Swiss-Prot, Entrez Gene and the Human Gene Nomenclature Committee (HGNC) drops to 18,690, indicating methodological differences in protein definitions and experimental existence support between sources. The Swiss-Prot and neXtProt evidence criteria include mass spectrometry peptide verification and also cross-references for antibody detection from the Human Protein Atlas. Notwithstanding, hundreds of Swiss-Prot entries are classified as non-coding biotypes by HGNC. The only inference that protein numbers might still rise comes from numerous reports of small ORF (smORF) discovery. However, while there have been recent cases of protein verifications from previous miss-annotation of non-coding RNA, very few have passed the Swiss-Prot curation and genome annotation thresholds. The post-genomic era has seen both advances in data generation and improvements in the human reference assembly. Notwithstanding, current numbers, while persistently discordant, show that the earlier yo-yoing has largely ceased. Given the importance to biology and biomedicine of defining the canonical human proteome, the task will need more collaborative inter-source curation combined with broader and deeper experimental confirmation in vivo and in vitro of proteins predicted in silico. The eventual closure could be well be below ~19,000.


Genetic examination of children suffering from cystic fibrosis.
CFTR protein (cystic fibrosis trans membrane conductance regulator) is expressed in multiple epithelial tissues, including upper and lower respiratory tracts, pancreas, sweat glands and gastrointestinal tract. More than 800 mutations and 100 polymorphic variants of DNA sequences were identified in patients with CF (Cystic fibrosis) and CFTR-diseases. In this study, genetic CFTR analysis of the children suffering from chronic lung disease (cystic fibrosis) is presented. They are treated and regularly controlled at the Pediatric hospital Sarajevo. CFTR analysis was done in 9 cases, 4 boys (44.4%) and 5 girls (55.55%). There are 3 children (33.3%) in the age group 1 to 3 years, 1 child (11.1%) in the age group 3 to 6 years, 3 children (33.3%) in the age group 6 to 9 years and 2 children (22.2%) in the age group 9 to 12 years. Genetic analysis was conducted at the Medical center for molecular biology, School of Medicine, Ljubljana. PCR method with PAGE and direct sequestration on ABI PRISM 31 was applied. The majority of children (7 children, i.e. 77.77%) had CFTR mutation Delta F 508 whilst one child had G542X mutation and one child R 1174 mutation. The purpose of this study is to emphasize the need for CFTR gene identification in the institutes of our country.


Development of mammalian artificial chromosomes for the treatment of genetic diseases: Sandhoff and Krabbe diseases.
Mammalian artificial chromosomes (MACs) are being developed as alternatives to viral vectors for gene therapy applications, as they allow for the introduction of large payloads of genetic information in a non-integrating, autonomously replicating format. One class of MACs, the satellite DNA-based artificial chromosome expression vehicle (ACE), is uniquely suited for gene therapy applications, in that it can be generated denovo in cells, along with being easily purified and readily transferred into a variety of recipient cell lines and primary cells. To facilitate the rapid engineering of ACEs, the ACE System was developed, permitting the efficient and reproducible loading of pre-existing ACEs with DNA sequences and/or target gene(s). As a result, the ACE System and ACEs are unique and versatile platforms for ex vivo gene therapy strategies that circumvent and alleviate existing safety and delivery limitations surrounding conventional gene therapy vectors. This review will focus on the status of MAC technologies and, in particular, the application of the ACE System towards an ex vivo gene therapy treatment of lysosomal storage diseases, specifically Sandhoff (MIM #268800) and Krabbe (MIM #245200) diseases.


Counting distance: Effects of egocentric distance on numerical perception.
Numerical value is long known to be associated with a variety of magnitude representations, such as size, time and space. The present study focused on the interactive relations of numerical magnitude with a spatial factor which is dominant in everyday vision and is often overlooked, namely, egocentric distance, or depth. We hypothesized that digits denoting large magnitudes are associated with large perceived distances, and vice versa. While the relations of numerical value and size have been long documented, effects of egocentric distance on numeral perception have been scarcely investigated, presumably due to the difficulty to disentangle size and depth factors within three-dimensional visual displays. The current study aimed to assess the potential linkage between egocentric distance and number magnitude, while neutralizing any perceived and/or physical size parameters of target digits. In Experiment 1, participants conducted a numeral size-classification task ('bigger or smaller than 5'), to which they responded with a near-to-body or a far-from-body key. Results revealed shorter responses for small than for large numbers when responded with a key positioned close to the body, and for large than small numbers when responded with a key positioned far from the body (regardless of hand-key mapping). Experiment 2 used verbal stimuli denoting near/remote concepts as irrelevant primes to target digits, further demonstrating a priming effect of conceived distance on numerical value processing. Collectively, our results suggest that distance magnitudes are associatively linked to numerical magnitudes and may affect digit processing independently of the effects of visual size.


[Scanning probe microscopy and medico-biological nanotechnology: history and prospects].
Literature data on the use of scanning probe microscopy for biological samples studying are presented. As an illustration, some results of the authors' investigations of the shark olfactory receptor surface, human lipoprotein of low density and human erythrocyte are given. Perspectives of the medico-biological development of nanotechnologies are evaluated.


Multiple alpha 2-adrenoceptor signalling pathways mediate pigment aggregation within melanophores.
It has previously been shown that alpha 2-adrenoceptors (alpha 2-ARs) mediate pigment granule (melanosome) aggregation in melanophores of the teleost fish Labrus ossifagus. The present investigation scrutinized the signalling mechanisms of melanosome aggregation induced by sympathetic nerve stimulation or by exogenous addition of alpha-AR agonists and cAMP analogues. The following was observed: i) nerve-induced melanosome aggregation was associated with a rapid decrease in the cAMP level; ii) noradrenaline or medetomidine (an alpha 2-AR agonist) caused melanosome aggregation and reduced the cAMP content; iii) RP-8-Cl-cAMP, a membrane-permeating inhibitor of protein kinase A induced melanosome aggregation; and iv) B-HT 920 (an alpha 2-AR agonist) and methoxamine (an alpha 1-AR agonist) induced melanosome aggregation, although they did not reduce cAMP. It has been suggested that in some teleost species alpha 1-ARs mediate melanosome aggregation by increasing the level of intracellular calcium. However, we found that the effect of methoxamine in melanophores from Labrus ossifagus could be blocked by yohimbine (an alpha 2-AR antagonist) but not by equimolar concentration of prazosin (an alpha 1-AR antagonist). Furthermore, 1 microM ionomycin (a calcium ionophore) did not induce melanosome aggregation. Our findings therefore do not indicate that alpha 1-ARs and/or an increase in intracellular calcium mediate melanosome aggregation in Labrus ossifagus. Our results suggest that alpha 2-AR-mediated melanosome aggregation is induced by multiple signalling pathways. One of these involves a reduction in cAMP, but none involves an increase in intracellular calcium.


PDGF receptors-mediators of autocrine tumor growth and regulators of tumor vasculature and stroma.
PDGFs and their cognate tyrosine kinase alpha- and beta-receptors are involved in multiple tumor-associated processes including autocrine growth stimulation of tumor cells, stimulation of tumor angiogenesis and recruitment and regulation of tumor fibroblasts. The recent development of clinically useful PDGF antagonists, like STI571/Glivec, has increased the interest in PDGF receptors as cancer drug targets. Autocrine PDGF receptor signaling occurs in certain malignancies characterized by mutational activation of PDGF or PDGF receptors, for instance, dermatofibrosaracoma protuberans, gastrointestinal stromal tumors, and hypereosinophilic syndrome. The roles of PDGF in regulation of tumor angiogenesis and tumor fibroblasts are more general, and probably occur in most common solid tumors. Concerning tumor angiogenesis recent studies have predominantly focused on the importance of PDGF receptor signaling for tumor pericyte recruitment. PDGF receptors in the tumor stroma have also attracted attention as interesting drug targets because of their function as regulators of tumor interstitial fluid pressure, tumor transvascular transport and tumor drug uptake. In summary, the improved understanding of the role of PDGF signaling in tumor biology, and the introduction of PDGF antagonists, has set the stage for a continued development of PDGF antagonists as novel cancer drugs.


DSA in acute gastrointestinal hemorrhage: clinical and in vitro studies.
Selective intraarterial digital subtraction angiography (DSA) was used to examine 37 patients with acute gastrointestinal (GI) tract bleeding. Conventional screen-film angiography was used as an adjunct to DSA when a larger field of view was needed (five patients) and when bowel motion prevented the acquisition of adequate image quality with DSA (two patients). Conventional angiography was also performed in all cases in which there were negative DSA examinations. DSA reduced the mean examination time considerably (20% reduction overall), especially for cases involving embolization therapy (35% reduction). DSA was especially valuable in the upper GI tract, where it was used to rapidly locate and/or assist in the embolization of bleeding sites in 19 of 20 patients with positive angiograms. There were 12 true-negative DSA examinations and one false-negative examination due to the limited field of view (9 inches [22.9 cm]). Bowel and respiratory motion were not important problems in the upper GI tract. In the lower GI tract, the usefulness of DSA was severely limited by the small field of view and the misregistration artifact caused by bowel motion. In an in vitro study, DSA and conventional angiography were compared as to their ability to depict several rates of extravasation of contrast material in a model of GI bleeding. DSA tended to be more sensitive for the detection of simulated extravasation (P less than .07).


Morphodynamic modeling of erodible laminar channels.
A two-dimensional model for the erosion generated by viscous free-surface flows, based on the shallow-water equations and the lubrication approximation, is presented. It has a family of self-similar solutions for straight erodible channels, with an aspect ratio that increases in time. It is also shown, through a simplified stability analysis, that a laminar river can generate various bar instabilities very similar to those observed in natural rivers. This theoretical similarity reflects the meandering and braiding tendencies of laminar rivers indicated by F. Métivier and P. Meunier [J. Hydrol. 27, 22 (2003)]. Finally, we propose a simple scenario for the transition between patterns observed in experimental erodible channels.


Tropic interactions between sympathetic nerves and vascular smooth muscle.
Explants of sympathetic ganglia from 3-9 day old rats were grown on collagen-coated coverslips in modified Rose chambers for 5 days, either alone or 2 mm away from explants of 6 week old rat caudal artery and aorta. Nerve fibre growth was stimulated on the side of the ganglion explant near the caudal artery explants but was not stimulated near the aorta. To determine the source of the nerve growth stimulation, explants of whole intact wall of the caudal artery, separated adventitia and medial layers, enzyme-dispersed cells, homogenized cells and medium pre-conditioned by caudal artery explants, were combined with ganglia. Explants of whole caudal artery and dispersed cells were also precultured prior to combination with ganglia. These combinations allowed analysis of the role of smooth muscle cells, existing nerve fibres, necrotic cells and connective tissue. Results suggested that degenerating nerve fibres within the blood vessels caused the increase in the number and the 'attraction' of the nerve fibres growing from the sympathetic ganglia. In contrast, both caudal artery and aorta from 3-9 day old rats caused stimulation of nerve fibre growth. Since these vessels were not yet innervated, the effect cannot be due to degenerating nerve terminals and a different mechanism must be involved.


Does an Annular Puncture Influence the Herniation Path?: An In Vitro Mechanical and Structural Investigation.
A study of mechanically induced herniation in punctured ovine discs followed by structural analysis. To investigate whether an annular puncture influences the path that herniation takes by providing direct passage for nucleus through the annulus and therefore whether it increases the risk of acute herniation from overload at the site of damage independent of any longer-term degeneration. Ten years after treatment with discography both degenerative changes and frequency of herniation have been shown to increase compared to untreated discs. Although the effect of an annular puncture over time has been widely investigated the question of whether it increases the risk of acute herniation has not been resolved. The posterolateral annuli of healthy ovine lumbar discs were punctured with either a 25-gauge (n = 8) or a larger 18-gauge (n = 8) needle and then compressed in a flexed posture of 10° until initial indications of failure. The entire volume of the disc was visually assessed for structural damage by obtaining progressive, full transverse cross-sections of its entire height thus exposing all regions of the disc. There was no association between the 25-gauge puncture and disc disruption and herniation. In contrast, nuclear material was observed to migrate through the 18-gauge needle puncture. Disruption of the lateral inner annulus was observed in 12 out of the 16 discs tested. The risk of acute herniation through the puncture site is dependent on the needle diameter used. Under the conditions employed the lateral inner annulus remains the site most vulnerable to disruption independent of the presence of a posterolateral puncture. N /A.


Microbead-based immunoassay for simultaneous detection of Shiga toxins and isolation of Escherichia coli O157 in foods.
Shiga toxin-producing Escherichia coli (STEC) is a significant foodborne pathogen with great economic consequences. There has been an increased food safety concern with this organism since outbreaks of human illnesses caused by this pathogen were first reported in 1982. Therefore, developing a reliable, sensitive, and rapid assay capable of detecting E. coli O157 and the main toxins produced by STEC (i.e., Shiga toxins 1 [Stx(1)] and 2 [Stx(2)]) will directly benefit regulatory agencies by minimizing analysis time. Here, we use Luminex technology to detect multiple analytes in a single 50-ml sample. Using commercially available monoclonal antibodies coupled to carboxylated magnetic microbeads, we developed an immunoassay capable of simultaneously serotyping E. coli O157 and detecting Stx(1) and/or Stx(2). The specificity and sensitivity of this immunoassay was tested against a collection of 34 E. coli isolates belonging to various O serogroups phenotypically different for Stx. The results were compared with microplate sandwich enzyme-linked immunosorbent assay (ELISA), and no cross-reactivity was observed for any of the monoclonal antibodies used. An increased sensitivity up to 1,000 times was observed in the microbead-based immunoassay when compared with the microplate sandwich ELISA. The results indicate that Luminex technology has the potential to simultaneously detect multiple targets without loss of specificity and/or sensitivity. A blind experiment was conducted with 48 samples of ground beef, lettuce, and milk spiked with ≤2 CFU/g E. coli. All the samples were correctly identified, with no false positives or false negatives. This microbead-based immunoassay could be extended to simultaneously detect additional foodborne pathogens and their toxic markers.


The structure and function of the 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase from Haemophilus influenzae.
The gene encoding the 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase of Haemophilus influenzae has been cloned and expressed in Escherichia coli. A complex of the purified protein with a substrate analog has been crystallized and its structure solved by multiple anomalous dispersion using phase information obtained from a single crystal of selenomethione-labeled protein. The enzyme folds into a four-stranded antiparallel beta-sheet flanked on one side by two alpha-helices and on the other by three consecutive alpha-helices, giving a novel beta1alpha1beta2beta3alpha2beta4alpha3alpha4alpha5 polypeptide topology. The three-dimensional structure of a binary complex has been refined at 2.1 A resolution. The location of the substrate analog and a sulfate ion gives important insight into the molecular mechanism of the enzyme.


A Strengths and Needs Assessment of Campus Sexual Assault Response.
Sexual violence and sexual assault (SA) disproportionately affect the young adult population aged 18-24 years, half of which are enrolled in higher education. Campuses are tasked with providing a safe space for students and enacting an appropriate response to campus SA (CSA). As part of a grant-funded program to strengthen SA nurse examiner (SANE) services to those who experience SA on college campuses, we conducted a campus needs assessment and a campus website review for key messaging and analyzed the responses to identify strengths and gaps in campus resources related to CSA. These findings were the basis of individualized Campus Community Summary reports shared with participants from each campus. Guided qualitative interviews with interdisciplinary stakeholders across seven campus communities and campus website reviews were used to identify campus resource strengths, gaps, and recommendations for improvement. Common strengths included awareness of campus programs and community services, leadership support for CSA activities, and community CSA response. Identified opportunities for improvement included strengthening interdisciplinary collaboration, solving transportation issues to obtain care, mental healthcare and advocacy services, awareness and resource messaging, and fighting stigma and bias. Campus websites reviews indicated gaps in key information about SANE care and how to access SANE services. Numerous gaps and opportunities to strengthen campus SA awareness and coordination and access to a timely response to SA were identified. Addressing these gaps is essential to ensure quality care and services for individuals who experience SA on college campuses.


Characteristics of spoilage-associated secondary cucumber fermentation.
Secondary fermentations during the bulk storage of fermented cucumbers can result in spoilage that causes a total loss of the fermented product, at an estimated cost of $6,000 to $15,000 per affected tank. Previous research has suggested that such fermentations are the result of microbiological utilization of lactic acid and the formation of acetic, butyric, and propionic acids. The objectives of this study were to characterize the chemical and environmental conditions associated with secondary cucumber fermentations and to isolate and characterize potential causative microorganisms. Both commercial spoilage samples and laboratory-reproduced secondary fermentations were evaluated. Potential causative agents were isolated based on morphological characteristics. Two yeasts, Pichia manshurica and Issatchenkia occidentalis, were identified and detected most commonly concomitantly with lactic acid utilization. In the presence of oxygen, yeast metabolic activities lead to lactic acid degradation, a small decline in the redox potential (E(h), Ag/AgCl, 3 M KCl) of the fermentation brines, and an increase in pH to levels at which bacteria other than the lactic acid bacteria responsible for the primary fermentation can grow and produce acetic, butyric, and propionic acids. Inhibition of these yeasts by allyl isothiocyanate (AITC) resulted in stabilization of the fermented medium, while the absence of the preservative resulted in the disappearance of lactic and acetic acids in a model system. Additionally, three Gram-positive bacteria, Lactobacillus buchneri, a Clostridium sp., and Pediococcus ethanolidurans, were identified as potentially relevant to different stages of the secondary fermentation. The unique opportunity to study commercial spoilage samples generated a better understanding of the microbiota and environmental conditions associated with secondary cucumber fermentations.


[Comparison of single-agent docetaxel versus docetaxel plus platinum combination agent in second-line treatment for advanced non-small cell lung cancer].
To compare the therapeutic efficacies and toxicities of single-agent docetaxel or docetaxel plus platinum combination agent in patients with advanced non-small cell lung cancer (NSCLC) so as to provide rationales for standard second-line chemotherapy. The clinical data from 152 patients with NSCLC who were admitted into Chest Hospital Affiliated to Shanghai Jiaotong University, from January 2004 to May 2008 were retrospectively analyzed. Forty patients were treated with single-agent docetaxel (single-agent group, 16 and 24 patients with stages IIIb and IV disease respectively; 32 patients with PS score 0-1 before treatment and 8 patients with PS score 2 before treatment), and 112 patients were treated with docetaxel plus platinum combination agent (combination group, 29 and 83 patients with stage IIIb and IV disease respectively; 98 patients with PS score 0-1 before treatment and 14 patients with PS score 2 before treatment). Primary end point was overall survival (OS), and secondary end point were progression-free survival time (PFS), disease control rate (DCR), one-year survival, and drug toxicity. Survival analysis was evaluated by Kaplan-Meier. Single factor analysis and Cox regression model were done to analyze the relationship between the influencing factors and the prognosis of disease. The median PFS of the single-agent group was 3.0 months, significantly shorter than that of the combination group (4.2 months, P = 0.048). However, no statistical differences were found in OS, DCR or one-year survival between the two groups (all P > 0.05). The most common grade 3 to 4 toxicities were leukopenia (32.5% for single-agent group, and 56.2% for combination group, P = 0.000), and gastro-intestinal toxicity (0 for single-agent group, and 4.5% for combination group, P = 0.000). Single factor analysis showed that the factors including radiotherapeutic history, operative history, PS score before treatment, clinical stage, and response to second-line treatment influenced the prognosis of NSCLC (all P < 0.05). Cox regression analysis demonstrated operative history (HR = 0.428, 95% CI: 0.261-0.701), PS score before treatment (HR = 1.919, 95% CI: 0.999-3.685), clinical stage (HR = 2.297, 95% CI: 1.427-3.696) and response to second-line treatment (HR = 0.318, 95% CI: 0.177-0.571) had effects on survival. To those well-selected patients, docetaxel plus platinum combination agent as the second-line treatment of advanced NSCLC significantly prolongs the progression-free survival. But such a regimen is more toxic and does not improve the response rate and overall survival.


Conformational analysis of chiral supramolecular aggregates: modeling the subtle difference between hydrogen and deuterium.
A detailed analysis of the conformational states of self-assembled, stereoselectively deuterated benzene-1,3,5-tricarboxamides ((S,S,S)-D-BTAs) reveals four different conformers for the supramolecular polymers. The relative amount of the conformers depends on the solvent structure and the temperature. With the help of a model, the thermodynamic parameters that characterize the different conformational states were quantified as well as the amount of the species that occur at different stages of the polymerization process. The results show that small changes in the stability between different types of conformers formed by (S,S,S)-D-BTAs—in the order of a few J mol(–1)—arise from the combination of interactions between the solvent/supramolecular aggregate, temperature, and solvent structure. While the introduction of a deuterium label allows to sensitively probe the solvophobic effects in the supramolecular aggregation, a rationalization of the observed effects on a molecular level is not yet straightforward but is proposed to result from subtle effects in the vibrational enthalpy and entropy terms of the isotope effect.


[Interaction of a substance based on phytoecdysteroids (BTK-8L) with components of fractionated chromatin from intact rat liver].
Results of the study of interaction of the phytoecdysteroid preparation (BTK-8L) with fractionated rat liver nuclear chromatin were described. It was shown that the interaction resulted in the "loosening" of the histone proteins structure both in active and repressed chromatin. At the same time the reparative DNA synthesis in the repressed fraction was stimulated. The Change revealed might be the basis of protective properties of BTK-8L as to chromatin at chloro and organophosphorus intoxications.


Psychodrama: an innovative way of improving self-awareness of nurses.
The aim of this educational session was to form a group interaction model for improving the self-awareness of nurses via psychodrama. The structured group interaction session was conducted three times, with three separate groups, during the 'Intensive Care Nursing' training programme at a teaching hospital in Ankara. An assessment was made using the written records and observations of the group director and co-directors and feedback from nurses regarding the session. The nurses were highly motivated, adapting readily to the learning environment and following the instructions without difficulty. They were able to describe their personal experience with a specific patient and also to identify the fundamental emotion engendered by that interaction. Their feedback regarding the session was favourable. The psychodrama technique helped the nurses to understand themselves, to explore the perspective of others and to make the connection between their own thoughts/feelings and those of their patients. Psychodrama can be an effective teaching tool in addressing the communication issues that arise in nursing.


Risk scoring systems for predicting preterm birth with the aim of reducing associated adverse outcomes.
Identification of pregnancies that are higher risk than average is important to allow the possibility of interventions aimed at preventing adverse outcomes like preterm birth. Many scoring systems designed to classify the risk of a number of poor pregnancy outcomes (e.g. perinatal mortality, low birthweight, and preterm birth) have been developed, but they have usually been introduced without evaluation of their utility and validity. To determine whether the use of a risk-screening tool designed to predict preterm birth (in combination with appropriate consequent interventions) reduces the incidence of preterm birth and very preterm birth, and associated adverse outcomes. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (December 2010), CENTRAL (The Cochrane Library 2010, Issue 4), MEDLINE (1966 to 17 December 2010), EMBASE (1974 to 17 December 2010), and CINAHL (1982 to 17 December 2010). All randomised or quasi-randomised (including cluster-randomised) or controlled clinical trials that compared the incidence of preterm birth between groups that used a risk scoring instrument to predict preterm birth with those who used an alternative instrument, or no instrument; or that compared the use of the same instrument at different gestations. The reports may have been published in peer reviewed or non-peer reviewed publications, or not published, and written in any language. All review authors planned to independently assess for inclusion all the potential studies we identified as a result of the search strategy. However, we identified no eligible studies. Extensive searching revealed no trials of the use of risk scoring systems to prevent preterm birth. The role of risk scoring systems in the prevention of preterm birth is unknown.There is a need for prospective studies that evaluate the use of a risk-screening tool designed to predict preterm birth (in combination with appropriate consequent interventions) to prevent preterm birth, including qualitative and/or quantitative evaluation of their impact on women's well-being. If these prove promising, they should be followed by an adequately powered, well-designed randomised controlled trial.


Extrusion of lupines with or without addition of reducing sugars: Effects on the formation of Maillard reaction compounds, partition of nitrogen and Nε-carboxymethyl-lysine, and performance of dairy cows.
The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ15N in plasma and δ15N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars.


Proteolytic activation and solubilization of endoplasmic-reticulum cyclic AMP phosphodiesterase activity.
Dithiothreitol led to the activation and solubilization of the cyclic nucleotide phosphodiesterase activities associated with the smooth and various rough subfractions of rat liver endoplasmic reticulum. The activity in each of the subfractions exhibited somewhat different time courses, and sensitivities to dithiothreitol concentration, in respect of their solubilization and activation. Both activation and solubilization by dithiothreitol could be blocked by either thiol proteinase inhibitors or excess bovine serum albumin. Freeze-thaw solubilization was not blocked by the thiol proteinase inhibitor antipain and did not lead to the activation of the enzyme. After dithiothreitol-induced solubilization, all of the enzymes exhibited non-linear Lineweaver-Burk plots indicative of apparent negative co-operativity. In contrast, after freeze-thaw solubilization the enzyme in the smooth-endoplasmic-reticulum-plus-Golgi fraction still obeys Michaelis kinetics, as does the membrane-bound enzyme. It is possible to mimic the action of dithiothreitol in solubilizing and activating the enzyme by limited proteolysis with trypsin. Triton X-100 is highly efficient at solubilizing these enzymes, yet has little effect on their activities. Charged detergents exhibit highly selective effects on the enzymes as regards their solubilization and activity expressed.


The potential clinical value of plasma biomarkers in Alzheimer's disease.
Many people with cognitive complaints or impairment never receive an accurate diagnosis of the underlying condition, potentially impacting their access to appropriate treatment. To address this unmet need, plasma biomarker tests are being developed for use in Alzheimer's disease (AD). Plasma biomarker tests span various stages of development, including in vitro diagnostic devices (or tests) (IVDs), laboratory-developed tests (LDTs) and research use only devices (or tests) (RUOs). Understanding the differences between each test type is important for appropriate implementation into the AD diagnostic pathway and care continuum. Authors reviewed scientific literature (PubMed, meeting abstracts and presentations, company press releases and websites) on AD plasma biomarkers. This article defines IVDs, LDTs, and RUOs, discusses potential clinical applications and highlights the steps necessary for their clinical implementation. Plasma biomarkers could revolutionize many areas of the AD diagnostic pathway and care continuum, but further research is needed. There is a need for a minimally invasive Alzheimer's disease (AD) diagnostic tool. AD plasma biomarker tests exist at various stages of commercial development. Understanding the development stage of a test is important for its appropriate use. Plasma biomarker tests could function as a triage tool to streamline AD diagnosis. Further steps remain before AD plasma biomarkers can be used routinely.


Identifying the determinants in the equatorial domain of Buchnera GroEL implicated in binding Potato leafroll virus.
Luteoviruses avoid degradation in the hemolymph of their aphid vector by interacting with a GroEL homolog from the aphid's primary endosymbiotic bacterium (Buchnera sp.). Mutational analysis of GroEL from the primary endosymbiont of Myzus persicae (MpB GroEL) revealed that the amino acids mediating binding of Potato leafroll virus (PLRV; Luteoviridae) are located within residues 9 to 19 and 427 to 457 of the N-terminal and C-terminal regions, respectively, of the discontinuous equatorial domain. Virus overlay assays with a series of overlapping synthetic decameric peptides and their derivatives demonstrated that R13, K15, L17, and R18 of the N-terminal region and R441 and R445 of the C-terminal region of the equatorial domain of GroEL are critical for PLRV binding. Replacement of R441 and R445 by alanine in full-length MpB GroEL and in MpB GroEL deletion mutants reduced but did not abolish PLRV binding. Alanine substitution of either R13 or K15 eliminated the PLRV-binding capacity of the other and those of L17 and R18. In the predicted tertiary structure of GroEL, the determinants mediating virus binding are juxtaposed in the equatorial plain.


A review of health resource tracking in developing countries.
Timely, reliable and complete information on financial resources in the health sector is critical for sound policy making and planning, particularly in developing countries where resources are both scarce and unpredictable. Health resource tracking has a long history and has seen renewed interest more recently as pressure has mounted to improve accountability for the attainment of the health Millennium Development Goals. We review the methods used to track health resources and recent experiences of their application, with a view to identifying the major challenges that must be overcome if data availability and reliability are to improve. At the country level, there have been important advances in the refinement of the National Health Accounts (NHA) methodology, which is now regarded as the international standard. Significant efforts have also been put into the development of methods to track disease-specific expenditures. However, NHA as a framework can do little to address the underlying problem of weak government public expenditure management and information systems that provide much of the raw data. The experience of institutionalizing NHA suggests progress has been uneven and there is a potential for stand-alone disease accounts to make the situation worse by undermining capacity and confusing technicians. Global level tracking of donor assistance to health relies to a large extent on the OECD's Creditor Reporting System. Despite improvements in its coverage and reliability, the demand for estimates of aid to control of specific diseases is resulting in multiple, uncoordinated data requests to donor agencies, placing additional workload on the providers of information. The emergence of budget support aid modalities poses a methodological challenge to health resource tracking, as such support is difficult to attribute to any particular sector or health programme. Attention should focus on improving underlying financial and information systems at the country level, which will facilitate more reliable and timely reporting of NHA estimates. Effective implementation of a framework to make donors more accountable to recipient countries and the international community will improve the availability of financial data on their activities.


[Electromagnetic articulography: a new method for studying the motor function of the velum palatinum].
The observation of articulatory movements, particularly of velar movements, is of considerable theoretical and practical interest in evaluating the velopharyngeal function of the cleft palate patient. A new electromagnetic device is presented for tracking the movements of the velum and the tongue simultaneously in correlation with the posterior pharyngeal wall and the hard palate in the midsagittal plane. An inductive measuring principle is used; a miniature transmitting coil is placed at the lower margin of the velum, another coil is used for recording the position of the posterior pharyngeal wall as well as the surface of the hard palate and the movements of the tongue dorsum. The measuring principle is described and the movements of the velum during speech production are demonstrated in cleft palate patients with different speech results.


Estimated cost of needlestick injuries for six major needled devices.
A major factor in the introduction of new products designed to decrease the risk of needlesticks to healthcare workers (HCWs) is whether the increased expense of a safer device is offset by the savings of preventing needlesticks. The itemized costs of needlestick injuries associated with six major needled devices were estimated and compared to the cost of the devices causing the injuries, based on 1988 dollars. Included was the cost of treatment, prophylaxis and employee health department personnel time. The average cost of needlestick injury was $405, with a narrow range of $390 to $456 for different devices. As a percent of the cost of the devices, needlesticks cost as little as 10% of the cost of the device, for the intravenous (IV) catheter, to as much as 457%, for needles used to connect IV lines. On the average, needlesticks cost 36% of the devices' cost. These data may be used to weigh the potential economic benefits of safer needle technology or other strategies intended to reduce the incidence of needlesticks.


The Leucine Catabolite and Dietary Supplement β-Hydroxy-β-Methyl Butyrate (HMB) as an Epigenetic Regulator in Muscle Progenitor Cells.
β-Hydroxy-β-Methyl Butyrate (HMB) is a natural catabolite of leucine deemed to play a role in amino acid signaling and the maintenance of lean muscle mass. Accordingly, HMB is used as a dietary supplement by sportsmen and has shown some clinical effectiveness in preventing muscle wasting in cancer and chronic lung disease, as well as in age-dependent sarcopenia. However, the molecular cascades underlying these beneficial effects are largely unknown. HMB bears a significant structural similarity with Butyrate and β-Hydroxybutyrate (βHB), two compounds recognized for important epigenetic and histone-marking activities in multiple cell types including muscle cells. We asked whether similar chromatin-modifying actions could be assigned to HMB as well. Exposure of murine C2C12 myoblasts to millimolar concentrations of HMB led to an increase in global histone acetylation, as monitored by anti-acetylated lysine immunoblotting, while preventing myotube differentiation. In these effects, HMB resembled, although with less potency, the histone deacetylase (HDAC) inhibitor Sodium Butyrate. However, initial studies did not confirm a direct inhibitory effect of HMB on HDACs in vitro. β-Hydroxybutyrate, a ketone body produced by the liver during starvation or intense exercise, has a modest effect on histone acetylation of C2C12 cells or in vitro HDAC inhibitor activities, and, unlike Butyrate and HMB, did not interfere with myotube formation in a myoblast differentiation assay. Instead, βHB dramatically increased lysine β-hydroxybutyrylation (Kbhb) of histone tails, an epigenetic mark associated with fasting responses and muscle catabolic states. However, when C2C12 cells were exposed to βHB in the presence of equimolar HMB this chromatin modification was drastically reduced, pointing to a role for HMB in attenuating ketosis-associated muscle wasting. In conclusion, while their mechanistic underpinnings remain to be clarified, these preliminary observations highlight novel and potentially important activities of HMB as an epigenetic regulator and βHB antagonist in muscle precursor cells, to be further explored in their biomedical implications.


Transparent and broadband absorption-diffusion-integrated low-scattering metamaterial by standing-up lattice.
In this paper, a transparent absorption-diffusion-integrated metamaterial (ADMM) based on standing-up lattice structure is proposed which takes full advantage of electromagnetic absorption and destructive interference simultaneously for the suppression of broadband backward scattering within a wide angular domain, especially for the lower-frequency scattering. The proposed ADMM is constituted by two kinds of rhombic and squared ITO lattices arranged in a pseudorandom distribution and then backed with ITO film. Calculation, simulation, and experimental measurement show that the proposed ADMM can achieve low scattering with normalized reflection less than 0.1 in the frequency band of 6.1-21.0GHz. In addition, owing to the standing-up lattice structure, the averaged optical transmittance of our ADMM reaches the optimal value of around 82.1% in the visible wavelength range of 380-780nm, promising an excellent optical transparency. The proposed comprehensive scheme provides an effective way to achieve broadband scattering suppression and high compatibility with optical transparency, enabling a wide range of applications in the window glass of stealth armament, electromagnetic compatibility facility and photovoltaic solar device.


Mouse models for studying the formation and propagation of prions.
Prions are self-propagating protein conformers that cause a variety of neurodegenerative disorders in humans and animals. Mouse models have played key roles in deciphering the biology of prions and in assessing candidate therapeutics. The development of transgenic mice that form prions spontaneously in the brain has advanced our understanding of sporadic and genetic prion diseases. Furthermore, the realization that many proteins can become prions has necessitated the development of mouse models for assessing the potential transmissibility of common neurodegenerative diseases. As the universe of prion diseases continues to expand, mouse models will remain crucial for interrogating these devastating illnesses.


Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering.
Previous work documented the capacity of dendritic cells (DC) to stimulate primary immune responses and to physically cluster with the responding lymphocytes. Rapid cell-cell aggregation assays were used here to study the interaction of DC and other types of APC with T lymphocytes. Graded doses of APC were sedimented with T cells that had been primed to alloantigens, soluble proteins, or lectin, and then labeled with carboxyfluorescein diacetate. The number of clustered T cells was measured after 10 min at 4 or 37 degrees C. At 4 degrees, binding was antigen-dependent and included greater than 50% of the added T cells. Clustering was mediated by all types of APC tested, including DC, macrophages, B lymphocytes, and fresh Langerhans cells, although DC were the most effective. Specificity was evident in the findings that alloreactive T lymphoblasts bound to allogeneic but not syngeneic APC; KLH- and OVA-reactive T cells bound to syngeneic APC in the presence of specific protein: and Con A blasts needed lectin to cluster. A 30 min pretreatment with chloroquine, a drug known to inhibit APC activity, markedly blocked the specific binding of alloreactive and protein-specific T blasts at 4 degrees C. Since Lyt-2- alloreactive blasts should specifically recognize Ia, presentation of Ia seems to be altered by chloroquine. Binding assays at 37 degrees C gave similar results to those performed at 4 degrees C, with one exception. When DC were used as APC, striking antigen-independent clustering occurred. DC could efficiently cluster primed T cells in the absence of alloantigen, soluble protein, or lectin. We suggest that antigen-independent binding contributes to the distinctive capacity of DC to prime T cells in the afferent limb of the immune response, whereas antigen-dependent binding between other APC and sensitized lymphocytes is critical in the efferent limb.


[Quantitative analysis of the ultrastructure of gastric mucocytes in the rat after prostaglandin treatment].
Effect of prostaglandin E2 (PGE2) on mucus secretion by mucocytes of the fundal part of the rat stomach has been studied electron microscopically, morphometrically and biochemically. PGE2 is found to stimulate synthesis of glycoproteins and their secretion by the mucocytes that results in increasing the relative volume of Golgi complex and in decreasing that of secretory granules. Actinomycin does not affect and cycloheximide blocks the stimulating activity of prostaglandin. This demonstrates that prostaglandin produces its action at the posttranscriptional (translational) level controlling mucus secretion of mucocytes. Owing to the results of the investigation and of those previously obtained on incorporation of 3H-pentagastrin and 3H-histamine into the cells of the mucous tunic, at least two mechanisms are supposed to control the gastric mucus secretion.


Losses of sexual dichromatism involve rapid changes in female plumage colors to match males in New World blackbirds.
Differences in coloration between the sexes (sexual dichromatism) can increase or decrease in a species through evolutionary changes in either or both sexes diverging or converging in their colors. Few previous studies, however, have examined the relative rates of such changes, particularly when dichromatism is lost. Using reflectance data from 37 species of the New World blackbird family (Icteridae), we compared evolutionary rates of plumage color change in males and females when dichromatism was either increasing (colors diverging) or decreasing (colors converging). Increases in dichromatism involved divergent changes in both sexes at approximately equal rates. Decreases in dichromatism, in contrast, involved changes in females to match male plumage colors that were significantly more rapid than any changes in males. Such dramatic changes in females show how selection can differ between the sexes. Moreover, these evolutionary patterns support the idea that losses of dimorphism involve genetic mechanisms that are already largely present in both sexes, whereas increases in dimorphism tend to involve the appearance of novel sex-specific traits, which evolve more slowly. Our results have broad implications for how sexual dimorphisms evolve.


A conserved Asn in transmembrane helix 7 is an on/off switch in the activation of the thyrotropin receptor.
The thyrotropin (TSH) receptor is an interesting model to study G protein-coupled receptor activation as many point mutations can significantly increase its basal activity. Here, we identified a molecular interaction between Asp(633) in transmembrane helix 6 (TM6) and Asn(674) in TM7 of the TSHr that is crucial to maintain the inactive state through conformational constraint of the Asn. We show that these residues are perfectly conserved in the glycohormone receptor family, except in one case, where they are exchanged, suggesting a direct interaction. Molecular modeling of the TSHr, based on the high resolution structure of rhodopsin, strongly favors this hypothesis. Our approach combining site-directed mutagenesis with molecular modeling shows that mutations disrupting this interaction, like the D633A mutation in TM6, lead to high constitutive activation. The strongly activating N674D (TM7) mutation, which in our modeling breaks the TM6-TM7 link, is reverted to wild type-like behavior by an additional D633N mutation (TM6), which would restore this link. Moreover, we show that the Asn of TM7 (conserved in most G protein-coupled receptors) is mandatory for ligand-induced cAMP accumulation, suggesting an active role of this residue in activation. In the TSHr, the conformation of this Asn residue of TM7 would be constrained, in the inactive state, by its Asp partner in TM6.


Bryostatin-1 stimulates the transcription of cyclooxygenase-2: evidence for an activator protein-1-dependent mechanism.
Bryostatin-1 (bryostatin) is a macrocyclic lactone derived from Bugula neritina, a marine bryozoan. On the basis of the strength of in vitro and animal studies, bryostatin is being investigated as a possible treatment for a variety of human malignancies. Severe myalgias are a common dose-limiting side effect. Because cyclooxygenase-2 (COX-2)-derived prostaglandins can cause pain, we investigated whether bryostatin induced COX-2. Bryostatin (1-10 nM) induced COX-2 mRNA, COX-2 protein, and prostaglandin biosynthesis. These effects were observed in macrophages as well as in a series of human cancer cell lines. Transient transfections localized the stimulatory effects of bryostatin to the cyclic AMP response element of the COX-2 promoter. Electrophoretic mobility shift assays and supershift experiments revealed a marked increase in the binding of activator protein-1 (AP-1)(c-Jun/c-Fos) to the cyclic AMP response element of the COX-2 promoter. Pharmacological and transient transfection studies indicated that bryostatin stimulated COX-2 transcription via the protein kinase C-->mitogen-activated protein kinase-->AP-1 pathway. All-trans-retinoic acid, a prototypic AP-1 antagonist, blocked bryostatin-mediated induction of COX-2. Taken together, these results suggest that bryostatin-mediated induction of COX-2 can help to explain the myalgias that are commonly associated with treatment. Moreover, it will be worthwhile to evaluate whether the addition of a selective COX-2 inhibitor can increase the antitumor activity of bryostatin.


Charge-storage mechanism of highly defective NiO nanostructures on carbon nanofibers in electrochemical supercapacitors.
An electrode composed of highly defective nickel oxide (NiO) nanostructures supported on carbon nanofibers (CNFs) and immersed in an Li+-based aqueous electrolyte is studied using Raman spectroscopy under dynamic polarization conditions to address the charge-storage phenomenon. By this operando technique, the formation of Li2SO4·H2O during the discharge process is verified. At the same time, we observed the phase transformation of NiO to NiOOH. The Ni(OH)2/NiOOH redox couple is responsible for the pseudocapacitive behavior with intercalation of cationic species in the different Ni structures. A 'substitutive solid-state redox reaction' is proposed to represent the amphoteric nature of the oxide, resulting in proton intercalation, while the insertion of Li+ occurs to a less extent. The electrode material exhibits outstanding stability with 98% coulombic efficiency after 10 000 charge-discharge cycles. The excellent electrode properties can be ascribed to a synergism between CNFs and NiO, where the carbon nanostructures ensured rapid electron transport from the hydrated nickel nanoparticles. The NiO@CNF composite material is a promising candidate for future applications in aqueous-based supercapacitors. DFT simulation elucidates that compressive stress and Ni-site displacement lead to a decrease up-to 3.5-fold on the electron density map located onto the Ni-atom, which promotes NiO/Ni(OH)2/NiOOH transition.


Cladosporium Species Recovered from Clinical Samples in the United States.
Cladosporium species are ubiquitous, saprobic, dematiaceous fungi, only infrequently associated with human and animal opportunistic infections. We have studied a large set of Cladosporium isolates recovered from clinical samples in the United States to ascertain the predominant species there in light of recent taxonomic changes in this genus and to determine whether some could possibly be rare potential pathogens. A total of 92 isolates were identified using phenotypic and molecular methods, which included sequence analysis of the internal transcribed spacer (ITS) region and a fragment of the large subunit (LSU) of the nuclear ribosomal DNA (rDNA), as well as fragments of the translation elongation factor 1 alpha (EF-1α) and actin (Act) genes. The most frequent species was Cladosporium halotolerans (14.8%), followed by C. tenuissimum (10.2%), C. subuliforme (5.7%), and C. pseudocladosporioides (4.5%). However, 39.8% of the isolates did not correspond to any known species and were deemed to comprise at least 17 new lineages for Cladosporium. The most frequent anatomic site of isolation was the respiratory tract (54.5%), followed by superficial (28.4%) and deep tissues and fluids (14.7%). Species of the two recently described cladosporiumlike genera Toxicocladosporium and Penidiella are reported for the first time from clinical samples. In vitro susceptibility testing of 92 isolates against nine antifungal drugs showed a variety of results but high activity overall for the azoles, echinocandins, and terbinafine.