Shape-shifting 3D protein microstructures with programmable directionality via quantitative nanoscale stiffness modulation.
The ability to shape-shift in response to a stimulus increases an organism's survivability in nature. Similarly, man-made dynamic and responsive "smart" microtechnology is crucial for the advancement of human technology. Here, 10-30 μm shape-changing 3D BSA protein hydrogel microstructures are fabricated with dynamic, quantitative, directional, and angle-resolved bending via two-photon photolithography. The controlled directional responsiveness is achieved by spatially controlling the cross-linking density of BSA at a nanometer lengthscale. Atomic force microscopy measurements of Young's moduli of structures indicate that increasing the laser writing distance at the z-axis from 100-500 nm decreases the modulus of the structure. Hence, through nanoscale modulation of the laser writing z-layer distance at the nanoscale, control over the cross-linking density is possible, allowing for the swelling extent of the microstructures to be quantified and controlled with high precision. This method of segmented moduli is applied within a single microstructure for the design of shape-shifting microstructures that exhibit stimulus-induced chirality, as well as for the fabrication of a free-standing 3D microtrap which is able to open and close in response to a pH change.


Immune-Guided Therapy of COVID-19.
Vaccination has been a game changer in our efforts to address the coronavirus disease 2019 (COVID-19) pandemic. However, the disease might still represent a clinical crisis for several more years, in part because of the inevitable emergence of variants capable of evading the preexisting immunity. Drugs affecting viral spread will help curtail transmission, but therapeutics are needed to treat the more severe cases requiring hospitalization. A deep analysis of the evolving immune landscape of COVID-19 suggests that understanding the molecular bases of the distinct clinical stages is paramount if we are to limit the burden of inflammation, which can lead to death in frail individuals, according to age, sex, and comorbidities. Different phases can be defined using immune biomarkers and need specific therapeutic approaches, tailored to the underlying immune contexture.


Mesenchymal stromal cell-derived CCL2 suppresses plasma cell immunoglobulin production via STAT3 inactivation and PAX5 induction.
We demonstrate that the secretome of mesenchymal stromal cells (MSCs) suppresses plasma cell (PC) immunoglobulin (Ig) production, induces plasmablast proliferation, and leads to interleukin-10-mediated blockade in vitro. We found that these effects are the result of MSC-derived CC chemokine ligands CCL2 and CCL7. More specifically, MSCs further processed these CC chemokines by the activity of matrix metalloproteinases (MMPs), leading to the generation of proteolytically processed antagonistic CCL2 variant. Neutralizing CCL2 or inhibiting MMP enzymatic activity abolished the PC-suppressive effect of MSCs. We also observed that MMP-processed CCL2 suppresses signal transducer and activator of transcription 3 (STAT3) activation in PC. As a result, the transcription factor PAX5 is induced, thus explaining the inhibition of Ig synthesis. The absence of inhibitory effects by MSC on the humoral response of CCR2(-/-) mice to xenoantigen suggests that MMP-cleaved CCL2/CCR2 interaction as well as downstream phosphatase activity is necessary for antagonistic effect. We tested syngeneic MSCs in hemophilic B6 mice with predeveloped antihuman factor VIII (hFVIII) antibodies and demonstrated a robust decrease in hFVIII-specific IgG levels. Thus, MSCs may play a role in modulating Ig production by PCs via MMP processing of CCL2 and may represent an appealing cell therapy approach for pathologic humoral responses.


Effect of Rosa damascena Mill. flower extract on rat ileum.
Rosa damascena flower is widely used for gastrointestinal (GI) disorders. However, its pharmacological action on isolated ileum has not been studied. In this research, the effect of extract of flower petals of R. damascena Mill. growing in Kashan, Iran, on ileum motility was investigated. Hydroalcoholic extract was prepared by percolation method. A section of rat ileum was suspended in an organ bath containing Tyrode's solution. The tissue was stimulated with electrical field stimulation (EFS), KCl and acetylcholine (ACh). The tissue was kept under 1g tension at 37°C and continuously gassed with O2. Effect of the R. damascena extract was studied on ileum contractions induced by EFS, KCl and ACh and compared with that of atropine. R. damascena extract (10-100 mg/ml) induced a contraction in rat isolated ileum while at 1mg bath concentration it had relaxant effect on rat ileum. Hydroalcoholic extract of R. damascena (1-8 mg/ml) concentration dependently inhibited ileum contraction induced by KCl (IC50=3.3 ± 0.9 mg/ml), ACh (IC50=1.4 ± 0.1 mg/ml) and EFS (IC50=1.5 ± 0.3 mg/ml). The vehicle had no significant effect on ileum contractions. From this experiment it was concluded that R. damascena extract at microgram concentrations had stimulatory effect on ileum smooth muscle. However, at milligram concentrations, it shows an inhibitory effect. This is most likely due to presence of different components in the extract. The stimulatory effect of the extract confirms its benefits for the treatment of constipation. Therefore, separation and identification of active components is recommended.


Prognostic value of neutrophil-to-lymphocyte ratio in urothelial carcinoma of the upper urinary tract and bladder: a systematic review and meta-analysis.
The neutrophil-to-lymphocyte ratio (NLR) is an inflammation marker that has prognostic value for various tumors, but its prognostic value in urothelial carcinoma (UC) remains controversial. This meta-analysis investigated the prognostic value of NLR in UC. A systematic search was performed on PubMed, ISI Web of Science, and Embase for studies focusing on the association between NLR and clinical features or prognosis of UC and published until November 2016. Prognostic outcomes and clinical features were collected and analyzed. A total of 11,538 patients from 32 studies were included in the meta-analysis. Increased pretreatment NLR predicted poor overall survival (hazard ratio [HR] = 1.72, 95% confidence interval [CI] = 1.45-2.05), progression free survival (HR = 1.68, 95% CI = 1.44-1.96), and cancer specific survival (HR = 1.64, 95% CI = 1.39-1.93) in all the patients. The increased pretreatment NLR was correlated with increased lymphovascular invasion (HR = 1.29, 95% CI = 1.17-1.43), high tumor T stage (HR = 1.25, 95% CI = 1.12-1.39), and tumor grade (HR = 1.07, 95% CI = 1.01-1.14) but not with lymph node involvement, carcinoma in situ, multifocality, or positive margin. Our meta-analysis indicated that NLR could predict the prognosis for UC and was associated with UC progression in terms of lymphovascular invasion, tumor T stage, and tumor grade.


Verbal processing deficits in schizophrenia.
The authors reported that a subgroup of schizophrenic patients performed well on a tone serial position task but was impaired on an auditory word serial position task (Wexler, Stevens, Bowers, Cerniak, & Goldman-Rakic, 1998). This study assessed 30 schizophrenic and 32 controls (matched for comparable tone discrimination) on 4 versions of the verbal serial position tasks and 2 tone serial position tasks. Patients performed poorly on all verbal tasks but performed comparably to controls when tones served as stimuli. Proactive interference and visual presentation further compounded the verbal deficits. Deficits persisted with pronounceable nonword stimuli. These findings provide evidence of specific deficits in language-related processing, although the authors could not rule out the possibility that the differential effects that were observed between the tone and word tasks, and particularly among the verbal tasks, may result from differing discriminating power of the different tests.


The chemokine CXCL1/growth related oncogene increases sodium currents and neuronal excitability in small diameter sensory neurons.
Altered Na+ channel expression, enhanced excitability, and spontaneous activity occur in nerve-injury and inflammatory models of pathological pain, through poorly understood mechanisms. The cytokine GRO/KC (growth related oncogene; CXCL1) shows strong, rapid upregulation in dorsal root ganglion in both nerve injury and inflammatory models. Neurons and glia express its receptor (CXCR2). CXCL1 has well-known effects on immune cells, but little is known about its direct effects on neurons. We report that GRO/KC incubation (1.5 nM, overnight) caused marked upregulation of Na+ currents in acutely isolated small diameter rat (adult) sensory neurons in vitro. In both IB4-positive and IB4-negative sensory neurons, TTX-resistant and TTX-sensitive currents increased 2- to 4 fold, without altered voltage dependence or kinetic changes. These effects required long exposures, and were completely blocked by co-incubation with protein synthesis inhibitor cycloheximide. Amplification of cDNA from the neuronal cultures showed that 3 Na channel isoforms were predominant both before and after GRO/KC treatment (Nav 1.1, 1.7, and 1.8). TTX-sensitive isoforms 1.1 and 1.7 significantly increased 2 - 3 fold after GRO/KC incubation, while 1.8 showed a trend towards increased expression. Current clamp experiments showed that GRO/KC caused a marked increase in excitability, including resting potential depolarization, decreased rheobase, and lower action potential threshold. Neurons acquired a striking ability to fire repetitively; IB4-positive cells also showed marked broadening of action potentials. Immunohistochemical labelling confirmed that the CXCR2 receptor was present in most neurons both in dissociated cells and in DRG sections, as previously shown for neurons in the CNS. Many studies on the role of chemokines in pain conditions have focused on their rapid and indirect effects on neurons, via release of inflammatory mediators from immune and glial cells. Our study suggests that GRO/KC may also have important pro-nociceptive effects via its direct actions on sensory neurons, and may induce long-term changes that involve protein synthesis.


Functional heterogeneity of osteopontin isoforms in non-small cell lung cancer.
Osteopontin (OPN) is a multifunctional protein with an important but poorly understood role in non-small cell lung cancer (NSCLC) pathogenesis. Moreover, the role of the three known mRNA isoforms (OPNa, OPNb, and OPNc) has not been reported. We hypothesize that OPN isoforms play different roles in determining the metastatic potential of NSCLC. We amplified mRNA for each OPN isoform in NSCLC tumors and matched normal lung. The functional impact of each isoform was evaluated by transfecting cDNA plasmids specific to each isoform into NSCLC cell lines and comparing behavior to empty vector controls in scratch closure, cell proliferation, soft-agar colony formation, and Matrigel invasion assays. Gene array was used to evaluate differences in downstream targets and was compared with a panel of markers for epithelial-mesenchymal transition (EMT). OPNa expression was increased in 91% of NSCLC tumors compared with matched lung. OPNa overexpression significantly increased activity in scratch closure, proliferation, soft-agar colony formation, and Matrigel invasion assays compared with controls in all cell lines. OPNb overexpression produced a less significant modulation of function. OPNc overexpression significantly decreased activity in proliferation, colony formation, and invasion assays compared with controls. Expression arrays revealed an increase in EMT with OPNa overexpression but not OPNc. Differences were validated by quantitative reverse transcriptase-polymerase chain reaction. Overexpression of the individual OPN isoforms in NSCLC results in divergent functional phenotypes. OPNa produced an aggressive phenotype, whereas OPNc produced a more indolent phenotype. Exon 4, which is transcribed in OPNa but absent in OPNc, may be central to this phenomenon and could serve as a target for isoform-specific inhibition of OPN in NSCLC.


Cost-effectiveness of a peer mentoring intervention to improve disease self-management practices and self-efficacy among African American women with systemic lupus erythematosus: analysis of the Peer Approaches to Lupus Self-management (PALS) pilot study.
The Peer Approaches to Lupus Self-management (PALS) program was developed as a peer mentoring tool to improve health behaviors, beliefs, and outcomes in African American women with systemic lupus erythematosus (SLE). This study aims to assess the cost of the PALS intervention and determine its effectiveness when compared to existing treatments. Peer mentors and mentees were paired on shared criteria such as life stage, marital status, or whether they were mothers. This 12-week program consisted of a weekly peer mentoring session by telephone. Cost of healthcare utilization was evaluated by assessing the healthcare costs pre- and post-intervention. Validated measures of quality of life, self-management, disease activity, depression, and anxiety were collected. Total direct program costs per participant were totaled and used to determine average per unit improvement in outcome measures. The benefit-cost ratio and pre- versus post-intervention hospital charges were examined. A total of 20 mentees and 7 mentors were enrolled in the PALS program. All PALS pairs completed 12 sessions lasting an average of 54 minutes. Mentees reported statistically significant decreases in patient-reported disease activity, depression, and anxiety, with improved trends in patient activation or patient engagement in their disease and management. The total cost per patient was $1291.50, which was $107.62 per patient per week. There was a savings of $23,417 per individual receiving the intervention with a benefit-cost ratio of 18.13 per patient. These findings suggest that the PALS intervention was effective in improving patient-level factors and was cost-effective. Future research will need to validate these findings in a larger sample.


THE AKR THYMIC ANTIGEN AND ITS DISTRIBUTION IN LEUKEMIAS AND NERVOUS TISSUES.
A clear-cut serological differentiation between AKR lymphocytes of thymic and non-thymic origin is reported: these two cell types are antigenically distinct. In newborn mice, the AKR thymic antigen was found at a high concentration only in thymus. In adult mice, the antigen was present at a high level in thymus, all nervous tissues tested, and some leukemias. It was present at much lower levels in lymph node lymphocytes, splenic lymphocytes, appendix, lung, and certain other leukemias, which appeared to be of non-thymic origin. The AKR thymic antigen was present at a high level in thymus and nervous tissues of RF mice, but was absent from thymocytes of sixteen other mouse strains. These sixteen strains possessed the C3HeB/Fe thymic antigen. The distribution of this antigen in neonatal and adult tissues of the strains tested was similar to that of the AKR thymic antigen in AKR mice. No exceptions were found. These results were obtained by use of immune cytolysis, and of a new method for the quantitative treatment of data from absorption experiments.


Association between human immunodeficiency virus infection and stiffness of the common carotid artery.
Individuals with human immunodeficiency virus (HIV) who use highly active antiretroviral therapy (HAART) may have an increased risk for cardiovascular-related events, although the underlying mechanism remains unclear. We tested the hypothesis that carotid arterial stiffness was higher among persons using HAART compared to HAART-naïve and HIV-uninfected persons. Between 2004 and 2006, we performed high-resolution B-mode ultrasound on 2789 HIV-infected and HIV-uninfected participants of the Women's Interagency HIV Study (1865 women) and the Multicenter AIDS Cohort Study (924 men) and determined carotid arterial distensibility, which is a direct measure of carotid arterial stiffness. We used generalized estimating equations to evaluate the association between distensibility and HIV infection, CD4(+) cell count, and exposure to HAART adjusted for demographic, behavioral, and clinical characteristics. In multivariable analysis, distensibility was 4.3% lower (95% confidence interval, -7.4% to -1.1%) among HIV-infected vs uninfected participants. Among HIV-infected participants with <200 CD4(+) cells, distensibility was 10.5% lower (95% confidence interval, -14.5% to -6.2%) than that among HIV-uninfected participants, and this effect did not differ significantly by cohort or race. Concurrent HAART use was independently associated with lower distensibility among Multicenter AIDS Cohort Study participants but not among Women's Interagency HIV Study participants. Our finding that advanced HIV-related immunosuppression was associated with increased carotid arterial stiffness independent from the effects of traditional atherosclerosis risk factors suggests that the etiologic mechanism underlying reports of an increased cardiovascular disease risk among HIV-infected individuals might involve HIV-related immunosuppression leading to vascular dysfunction and arterial stiffening.


Perturbational treatment of spin-orbit coupling for generally applicable high-level multi-reference methods.
An efficient perturbational treatment of spin-orbit coupling within the framework of high-level multi-reference techniques has been implemented in the most recent version of the Columbus quantum chemistry package, extending the existing fully variational two-component (2c) multi-reference configuration interaction singles and doubles (MRCISD) method. The proposed scheme follows related implementations of quasi-degenerate perturbation theory (QDPT) model space techniques. Our model space is built either from uncontracted, large-scale scalar relativistic MRCISD wavefunctions or based on the scalar-relativistic solutions of the linear-response-theory-based multi-configurational averaged quadratic coupled cluster method (LRT-MRAQCC). The latter approach allows for a consistent, approximatively size-consistent and size-extensive treatment of spin-orbit coupling. The approach is described in detail and compared to a number of related techniques. The inherent accuracy of the QDPT approach is validated by comparing cuts of the potential energy surfaces of acrolein and its S, Se, and Te analoga with the corresponding data obtained from matching fully variational spin-orbit MRCISD calculations. The conceptual availability of approximate analytic gradients with respect to geometrical displacements is an attractive feature of the 2c-QDPT-MRCISD and 2c-QDPT-LRT-MRAQCC methods for structure optimization and ab inito molecular dynamics simulations.


Hospital Opioid Requirements Following Continuation Versus Discontinuation of Buprenorphine for Addiction - A Retrospective Cohort Study.
Evidence guiding inpatient management of buprenorphine is lacking-this retrospective cohort study evaluated the clinical impact of hospital continuation versus discontinuation of buprenorphine at an academic medical center. The primary outcome was inpatient oral morphine equivalents (OME). Secondary outcomes included patient pain levels, functional assessment, and hospital length of stay. One hundred thirty-one patients (74 continued buprenorphine, 57 discontinued) were included in the analysis. Median OME were significantly lower among patients continued on buprenorphine versus discontinued (11 mg vs 103 mg, p < 0.001), as was maximum 24-hour opioid utilization (60 mg vs 240 mg, p < 0.001) and 24-hour pre-discharge utilization (10 mg vs 128 mg, p < 0.001). Median pain levels were similar between groups at the time of admission (8 in each group, p = 0.48), discharge (7 in each group, p = 0.26), and over the first 7 days of hospitalization (7 vs 8, p = 0.08). Hospital length of stay was similar between groups (5 days in each group, p > 0.99). Failure to reinitiate buprenorphine occurred in 31/57 patients (54.4%) in the discontinuation group. Hospital buprenorphine continuation is associated with reduced opioid requirements, while not significantly impacting pain levels, functionality, or length of admission. Failure to reinitiate buprenorphine was common and may have negative implications for addiction treatment.


Stereochemical considerations of the actions of some psychotropic drugs.
Stereoisomers have been used as tools to investigate the role of spatial (3-dimensional) features in neurochemical and behavioral effects of psychotropic drugs. Studies using enantiomers (optical isomers), geometrical isomers and semi-rigid molecules have shown that spatial features play a role in effects of antidepressant, antipsychotic, antiparkinsonian, and antianxiety drugs, as well as stimulants and psychotomimetics. An understanding of the role of spatial features in effects of psychotropic drugs might aid in reducing their side effects and in enhancing their therapeutic efficacy.


Intima-media thickness of the carotid arteries is related to serum osteoprotegerin levels in healthy postmenopausal women.
Osteoprotegerin (OPG) regulates bone mass by inhibiting osteoclast differentiation and activation, and also plays a role in vascular calcification. The objective of this study was to evaluate the relationship between serum OPG levels, and carotid artery intima-media thickness (IMT) and carotid plaque formation in healthy postmenopausal women. We recruited 68 healthy postmenopausal women for the study. Carotid plaque presence and IMT were evaluated by high resolution B-mode ultrasound. IMT was positively correlated with presence of plaque, age, menopause age and OPG, and inversely correlated with Apolipoprotein A1 (Apo A1). Serum OPG level was positively correlated with IMT (r = 0.366; p < 0.003) and age (r = 0.324; p < 0.008), and negatively correlated with Apo A1 (r = -0.481; p < 0.0001). We did not observe any significant relation between plaque occurrence and levels of serum OPG. In regression analysis OPG (p < 0.02) and menopause age (p < 0.05) were independent risk factors for IMT, and age (p < 0.05) and IMT (p < 0.05) were independent risk factors for plaque formation. Although the role of OPG in the vascular biology is poorly understood, our results suggest that elevated levels of serum OPG is associated with IMT and may play a role in the pathogenesis of atherosclerotic disease.


Association of the inflammatory balance of diet and lifestyle with colorectal cancer among Korean adults: a case-control study.
Dietary and lifestyle exposures may affect the risk of colorectal cancer (CRC) by promoting chronic inflammation. Therefore, we assessed the separate and joint associations of dietary and lifestyle inflammation scores (DIS and LIS, respectively) with CRC. Data from 919 pathologically confirmed CRC cases and 1,846 age- and sex-matched controls recruited at the National Cancer Center Korea were analyzed. We calculated the DIS and LIS, which characterize the collective contributions of 19 dietary and 4 lifestyle factors, respectively, to systemic inflammation by applying weights based on high-sensitivity C-reactive protein. A higher score represented a higher balance of pro- to anti-inflammatory exposures. Unconditional logistic regression models were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for CRC risk compared across the DIS and LIS tertile categories, with the lowest tertile as the reference group. The highest DIS tertile had significantly increased odds of having CRC (OR, 2.65; 95% CI, 2.10 to 3.36), and the odds increased with increasing DIS. The highest LIS tertile group had 1.28-fold higher odds of having CRC (95% CI, 1.03 to 1.58). In the cross-classification analysis, the odds of having CRC increased as the DIS and LIS jointly increased until the DIS reached the highest tertile, where the risk was very high (3-fold or more) regardless of the LIS. In conclusion, a higher balance of pro-inflammatory relative to anti-inflammatory dietary and lifestyle factors, especially dietary factors, was associated with higher CRC risk among Korean adults.


The amino acid sequence of the B-protein of bacteriophage ZJ-2.
1. Bacteriophage ZJ-2 is a filamentous male-specific coliphage. Methods are given for its preparation and separation into nucleic acid and the major coat-protein component (B-protein). 2. The determination of the amino acid sequence of the B-protein of bacteriophage ZJ-2 is described. The principal features of this aspect of the work were the special procedures dictated by the insolubility of the protein and the extreme insolubility of its fragments. The sequence is compared with that obtained by other workers for the coat protein of the related bacteriophage fd.


Cocrystal Structures of Antibody N60-i3 and Antibody JR4 in Complex with gp120 Define More Cluster A Epitopes Involved in Effective Antibody-Dependent Effector Function against HIV-1.
Accumulating evidence indicates a role for Fc receptor (FcR)-mediated effector functions of antibodies, including antibody-dependent cell-mediated cytotoxicity (ADCC), in prevention of human immunodeficiency virus type 1 (HIV-1) acquisition and in postinfection control of viremia. Consequently, an understanding of the molecular basis for Env epitopes that constitute effective ADCC targets is of fundamental interest for humoral anti-HIV-1 immunity and for HIV-1 vaccine design. A substantial portion of FcR effector function of potentially protective anti-HIV-1 antibodies is directed toward nonneutralizing, transitional, CD4-inducible (CD4i) epitopes associated with the gp41-reactive region of gp120 (cluster A epitopes). Our previous studies defined the A32-like epitope within the cluster A region and mapped it to the highly conserved and mobile layers 1 and 2 of the gp120 inner domain within the C1-C2 regions of gp120. Here, we elucidate additional cluster A epitope structures, including an A32-like epitope, recognized by human monoclonal antibody (MAb) N60-i3, and a hybrid A32-C11-like epitope, recognized by rhesus macaque MAb JR4. These studies define for the first time a hybrid A32-C11-like epitope and map it to elements of both the A32-like subregion and the seven-layered β-sheet of the gp41-interactive region of gp120. These studies provide additional evidence that effective antibody-dependent effector function in the cluster A region depends on precise epitope targeting--a combination of epitope footprint and mode of antibody attachment. All together these findings help further an understanding of how cluster A epitopes are targeted by humoral responses. HIV/AIDS has claimed the lives of over 30 million people. Although antiretroviral drugs can control viral replication, no vaccine has yet been developed to prevent the spread of the disease. Studies of natural HIV-1 infection, simian immunodeficiency virus (SIV)- or simian-human immunodeficiency virus (SHIV)-infected nonhuman primates (NHPs), and HIV-1-infected humanized mouse models, passive transfer studies in infants born to HIV-infected mothers, and the RV144 clinical trial have linked FcR-mediated effector functions of anti-HIV-1 antibodies with postinfection control of viremia and/or blocking viral acquisition. With this report we provide additional definition of the molecular determinants for Env antigen engagement which lead to effective antibody-dependent effector function directed to the nonneutralizing CD4-dependent epitopes in the gp41-reactive region of gp120. These findings have important implications for the development of an effective HIV-1 vaccine.


Surface nanoscale axial photonics: robust fabrication of high-quality-factor microresonators.
Recently introduced surface nanoscale axial photonics (SNAP) makes it possible to fabricate high-Q-factor microresonators and other photonic microdevices by dramatically small deformation of the optical fiber surface. To become a practical and robust technology, the SNAP platform requires methods enabling reproducible modification of the optical fiber radius at nanoscale. In this Letter, we demonstrate superaccurate fabrication of high-Q-factor microresonators by nanoscale modification of the optical fiber radius and refractive index using CO2 laser and UV excimer laser beam exposures. The achieved fabrication accuracy is better than 2 Å in variation of the effective fiber radius.


Evapotranspiration and favorable growing degree-days are key to tree height growth and ecosystem functioning: Meta-analyses of Pacific Northwest historical data.
While temperature and precipitation comprise important ecological filtering for native ranges of forest trees and are predisposing factors underlying forest ecosystem dynamics, the extent and severity of drought raises reasonable concerns for carbon storage and species diversity. Based on historical data from common garden experiments across the Pacific Northwest region, we developed non-linear niche models for height-growth trajectories of conifer trees at the sapling stage using annual or seasonal climatic variables. The correlations between virtual tree height for each locality and ecosystem functions were respectively assessed. Best-fitted models were composed of two distinct components: evapotranspiration and the degree-days disparity for temperature regimes between 5 °C and 18 °C (effective temperature sum and growth temperature, respectively). Tree height prediction for adaptive generalists (e.g., Pinus monticola, Thuja plicata) had smaller residuals than for specialists (e.g., Pinus contorta, Pseudotsuga menziesii), albeit a potential confounding factor - tree age. Discernably, there were linearly positive patterns between tree height growth and ecosystem functions (productivity, biomass and species diversity). Additionally, there was a minor effect of tree diversity on height growth in coniferous forests. This study uncovers the implication of key ecological filtering and increases our integrated understanding of how environmental cues affect tree stand growth, species dominance and ecosystem functions.


Predictors of cycling in college students.
To (1) assess cycling-related questions that have been added to the electronic version of the American College Health Association National College Health Assessment II (ACHA-NCHA), (2) examine cycling prevalence, and (3) identify predictors of cycling in college students. Predominately female (69%), undergraduate (89%), and white (85%) students (N = 949) from a large, urban, northwestern, bicycle-friendly university completed the electronic version of the ACHA-NCHA II. Thirty cycling-related questions were added to the ACHA-NCHA II and a subsample of questions was analyzed. Cycling questions added to the ACHA-NCHA II scale were reliable and valid, based on the psychometric data analysis. More than half (59%) of this sample cycled; of those, 58% cycled for transportation and 44% for recreation. Facilitators and barriers to cycling were different for cycling in general and cycling for transportation. Cycling questions added to the ACHA-NCHA II can be utilized to enhance knowledge relative to cycling on college campuses.


Biogenic and Synthetic Peptides with Oppositely Charged Amino Acids as Binding Sites for Mineralization.
Proteins regulate diverse biological processes by the specific interaction with, e.g., nucleic acids, proteins and inorganic molecules. The generation of inorganic hybrid materials, such as shell formation in mollusks, is a protein-controlled mineralization process. Moreover, inorganic-binding peptides are attractive for the bioinspired mineralization of non-natural inorganic functional materials for technical applications. However, it is still challenging to identify mineral-binding peptide motifs from biological systems as well as for technical systems. Here, three complementary approaches were combined to analyze protein motifs consisting of alternating positively and negatively charged amino acids: (i) the screening of natural biomineralization proteins; (ii) the selection of inorganic-binding peptides derived from phage display; and (iii) the mineralization of tobacco mosaic virus (TMV)-based templates. A respective peptide motif displayed on the TMV surface had a major impact on the SiO₂ mineralization. In addition, similar motifs were found in zinc oxide- and zirconia-binding peptides indicating a general binding feature. The comparative analysis presented here raises new questions regarding whether or not there is a common design principle based on acidic and basic amino acids for peptides interacting with minerals.


Candida auris: A Decade of Understanding of an Enigmatic Pathogenic Yeast.
Candida auris is an enigmatic yeast that continues to stimulate interest within the mycology community due its rapid and simultaneous emergence of distinct clades. In the last decade, almost 400 manuscripts have contributed to our understanding of this pathogenic yeast. With dynamic epidemiology, elevated resistance levels and an indication of conserved and unique pathogenic traits, it is unsurprising that it continues to cause clinical concern. This mini-review aims to summarise some of the key attributes of his remarkable pathogenic yeast.


An atypical pneumococcal arthritis.
Bone and joint infections due to Streptococcus pneumoniae usually occur in patients who are immunocompromised, and involve one site. The unique case of a 49-year-old immunocompetent man, with an unremarkable medical history, with septicaemia and polyarticular septic arthritis involving the shoulder and knee and with cervical spondylodiscitis due to S pneumoniae, is described. In this case, S pneumoniae probably originated from the gingiva, which is commonly colonised in children and adults. S pneumoniae should be considered routinely when facing bone and joint infections, and multiple locations should be carefully sought owing to the possible lack of symptoms.


Protective effect of arctiin against Toxoplasma gondii HSP70-induced allergic acute liver injury by disrupting the TLR4-mediated activation of cytosolic phospholipase A2 and platelet-activating factor.
Toxoplasma gondii (T. gondii)-derived heat shock protein 70 (T.g.HSP70) is a toxic protein that downregulates host defense responses against T. gondii infection. T.g.HSP70 was proven to induce fatal anaphylaxis in T. gondii infected mice through cytosolic phospholipase A2 (cPLA2) activated-platelet-activating factor (PAF) production via Toll-like receptor 4 (TLR4)-mediated signaling. In this study, we investigated the effect of arctiin (ARC; a major lignan compound of Fructus arctii) on allergic liver injury using T.g.HSP70-stimulated murine liver cell line (NCTC 1469) and a mouse model of T. gondii infection. Localized surface plasmon resonance, ELISA, western blotting, co-immunoprecipitation, and immunofluorescence were used to investigate the underlying mechanisms of action of ARC on T. gondii-induced allergic acute liver injury. The results showed that ARC suppressed the T.g.HSP70-induced allergic liver injury in a dose-dependent manner. ARC could directly bind to T.g.HSP70 or TLR4, interfering with the interaction between these two factors, and inhibiting activation of the TLR4/mitogen-activated protein kinase/nuclear factor-kappa B signaling, thereby inhibiting the overproduction of cPLA2, PAF, and interferon-γ. This result suggested that ARC ameliorates T.g.HSP70-induced allergic acute liver injury by disrupting the TLR4-mediated activation of inflammatory mediators, providing a theoretical basis for ARC therapy to improve T.g.HSP70-induced allergic liver injury.


Pharyngeal pressure generation during tongue-hold swallows across age groups.
To compare the effects of the tongue-hold swallowing maneuver on pharyngeal pressure generation in healthy young and elderly research volunteers. Sixty-eight healthy research volunteers (young, n = 34, mean age = 26.8 years, SD = 5.5; elderly, n = 34, mean age = 72.6 years, SD = 4.8; sex equally represented) performed 5 noneffortful saliva swallows and 5 tongue-hold swallows each. Amplitude and duration of pharyngeal pressure were investigated during both swallowing conditions with solid-state pharyngeal manometry at the level of the oropharynx, hypopharynx, and upper esophageal sphincter (UES). At both pharyngeal levels, tongue-hold swallows produced lower peak pressure compared with saliva swallows. During tongue-hold swallows, UES relaxation pressure was increased in the elders, whereas the younger group displayed a trend toward reduced relaxation pressure. Elderly individuals produced pressure longer during control swallows in the oropharynx and hypopharynx than young individuals. The tongue-hold maneuver affects oropharyngeal and hypopharyngeal pressure in the young and elders in similar ways, whereas effects on UES peak relaxation pressure differ between age groups. Reduced pharyngeal peak pressure and increased UES relaxation pressure underscore the notion that tongue-hold swallows should not be performed when bolus is present. Long-term training effects remain to be investigated.


GRADE concept 6: a novel application of external indirect evidence into GRADE ratings of evidence certainty in network meta-analysis.
We describe how consideration of external evidence may play an important role in judging certainty in the process of establishing the certainty of the evidence. Our example is a network meta-analysis (NMA) addressing treatment for Ebola virus disease, which informed a World Health Organization guideline. Through Grading of Recommendations Assessment, Development, and Evaluations (GRADE) project group iterative online, in-person and email discussions, we developed this GRADE concept and obtained approval from the GRADE working group. Using the null as a threshold, we rated our certainty for network estimates in mortality, including consideration of evidence external to the NMA (i.e., did not meet eligibility criteria) and formal logical construction. Based on the existing GRADE guidance, we rated the network estimate for one indirect comparison as low certainty. The formal logical construction that lead us reevaluate the certainty of the evidence is as follows: if A is superior to B, and B is not inferior to C, then A must be superior to C. After considering the logic and the external indirect evidence, we concluded at least moderate certainty for the comparison. Systematic review authors and guideline developers should apply the fundamental logical construction for indirect comparisons and consider compelling external evidence in NMA certainty ratings.


Drift and shape-new insights into human immunity against influenza virus neuraminidase.
Influenza virus hemagglutinin mediates infection by binding sialic acids, whereas neuraminidase cleaves sialic acids to release progeny virions. Both are targets of protective antibodies, but influenza vaccine strain selection and antigen dose are based on hemagglutinin alone. Virus characterization using first infection ferret sera indicates that escape from hemagglutination inhibiting (HI) antibodies occurs more frequently and is not coordinated with escape from neuraminidase inhibiting (NI) antibodies. A key question addressed by Daulagala et al. (P. Daulagala, B. R. Mann, K. Leung, E. H. Y. Lau, et al., mBio 14:e00084-23, 2023, https://doi.org/10.1128/mbio.00084-23) is how this translates to humans who encounter multiple influenza viruses throughout life. Their cross-sectional study, using sera from a wide age range of participants and H1N1 viruses spanning 1977-2015, indicates that NI antibodies are more broadly cross-reactive than HI antibodies. Both HI and NI titers were highest against strains encountered in childhood indicating that both are shaped by priming exposures. The study further supports the development of NA-optimized vaccines.


Cutting edge: the Th1 response inhibits the generation of peripheral regulatory T cells.
The possibility that effector T cells can be converted into forkhead box P3(+) regulatory T cells (Tregs) has potential therapeutic implications. To analyze the relationship between Th1 effectors and Tregs, we have used a model of systemic autoimmunity in which both effector and Tregs arise from a single population specific for a transgene-encoded systemic protein. In vitro, the presence of IFN-gamma inhibits Treg generation during activation. Using IFN-gamma reporter mice, we demonstrate that IFN-gamma-producing cells tend not to develop into Tregs, and Th1 priming of T cells prior to cell transfer limits the number of forkhead box P3(+) T cells generated in vivo. Moreover, transfer of IFN-gamma(-/-) or STAT1(-/-) T cells resulted in an increase in the number of Tregs. These data support a role for Th1 effector molecules and transcription factors in the control of peripheral Treg generation and demonstrates the limited plasticity of Th1 populations.


[Torsional tension in metaphase chromosome DNA].
The torsional tension in DNA of isolated metaphase chromosomes from murine fibroblasts was measured by the microfluorescent method. The method is based on the ability of a fluorescent dye ethidium bromide to compensate for the negative torsional tension in topologically closed DNA by intercalation between DNA base pairs. The value of the relative twist difference delta Tw/Tw = -0.1 was found in a bulk (about 3/4) of unconstrained chromosomal DNA. In interphase nuclei, the torsionally stressed DNA comprises about 15%, with value of delta Tw/Tw = -0.075. We suppose that the tension in chromosomal DNA was created in the prophase stage of mitosis by condensines, the drivers of chromosomal condensation.


A comparison of murine and human alveolar macrophage responses to urban particulate matter.
There is increasing evidence linking mortality, increased asthma morbidity, and other respiratory disorders to increases in fine airborne particulate matter (PM) concentrations. However, there are only limited data dealing with the biological mechanisms that ultimately lead to the reported health effects. Rodents are frequently used as an animal model to help elucidate the mechanisms of toxicity that may provide clues for the understanding of PM toxicity in humans; however, the relationships between murine and human PM toxicity have not been established. PM is known to target the pulmonary epithelium and resident alveolar macrophages (AM). PM can initiate cytotoxic effects on the AM including apoptosis and necrosis, depending on the particle concentration, which may be central to the pathological effects just described. This study examined AM apoptosis and necrosis initiated by PM in AM from humans and BALB/c mice in an in vitro exposure model. Freshly isolated AM from human volunteers were incubated with seven different residual fractions of PM1648 derived from organic solvent extractions, high-temperature heating and acid digestions that change the surface characteristics of the original PM. These results were compared to the analogous murine experiments. The results suggested that, at the same concentration of PM, the trend of toxicity and the posttreatment effects observed in BALB/c and human AM have a similar pattern. Altering the surface chemistry by removal of one or more PM components, such as through the various treatments conducted in this study, is sufficient to alter PM bioactivity in both human and murine AM in a similar manner. In addition, the human and murine models were compared with regard to in vitro cytotoxicity using PM(2.5) particles. The cytotoxic PM(2.5) effects were identical in both human and mouse models. Regression analysis revealed that the BALB/c mouse is a suitable model for PM cytotoxicity of AM as it is a good predictive model for the human AM responses.


Therapeutic effects of human multilineage-differentiating stress enduring (MUSE) cell transplantation into infarct brain of mice.
Bone marrow stromal cells (BMSCs) are heterogeneous and their therapeutic effect is pleiotropic. Multilineage-differentiating stress enduring (Muse) cells are recently identified to comprise several percentages of BMSCs, being able to differentiate into triploblastic lineages including neuronal cells and act as tissue repair cells. This study was aimed to clarify how Muse and non-Muse cells in BMSCs contribute to functional recovery after ischemic stroke. Human BMSCs were separated into stage specific embryonic antigen-3-positive Muse cells and -negative non-Muse cells. Immunodeficient mice were subjected to permanent middle cerebral artery occlusion and received transplantation of vehicle, Muse, non-Muse or BMSCs (2.5×104 cells) into the ipsilateral striatum 7 days later. Motor function recovery in BMSC and non-Muse groups became apparent at 21 days after transplantation, but reached the plateau thereafter. In Muse group, functional recovery was not observed for up to 28 days post-transplantation, but became apparent at 35 days post-transplantation. On immunohistochemistry, only Muse cells were integrated into peri-infarct cortex and differentiate into Tuj-1- and NeuN-expressing cells, while negligible number of BMSCs and non-Muse cells remained in the peri-infarct area at 42 days post-transplantation. These findings strongly suggest that Muse cells and non-Muse cells may contribute differently to tissue regeneration and functional recovery. Muse cells may be more responsible for replacement of the lost neurons through their integration into the peri-infarct cortex and spontaneous differentiation into neuronal marker-positive cells. Non-Muse cells do not remain in the host brain and may exhibit trophic effects rather than cell replacement.


Maternal Investment Fosters Male but Not Female Social Interactions with Other Group Members in Immature Wild Spider Monkeys (Ateles geoffroyi).
In several species, individuals form long-lasting social relationships with other group members, which provide them with important fitness benefits. In primates, patterns of social relationships are known to differ between sexes, but little is known about how these differences emerge through development or the role that mothers might have in this process. Here, we investigated how sex differences in social behaviour emerge during the first six years of primate life and how sex-biased maternal investment can foster immatures' social development and social interaction with other group members. For this purpose, we observed 20 males and females aged between zero and six years in a wild group of spider monkeys (Ateles geoffroyi) that was male-philopatric and, therefore, expected to show sex-biased maternal investment. Our results showed no sex difference in the social development of offspring with regards to body contact and grooming, but the probability of play was rather constant throughout age for females, whereas, for males, it became higher than females around two years of age, peaking between three and four years of age. Moreover, we found differences between female and male immatures in the importance of maternal investment (which included the time mothers spent nursing, carrying, grooming, touching and playing with their offspring) for their social integration in the natal group. In particular, maternal investment increased the probability of playing with other group members for sons, but not for daughters. Our findings suggest that mothers, through sex-biased maternal investment, might have a crucial function in the social development of spider monkeys, fostering the abilities that young offspring need to thrive as adults. By shedding light on maternal investment and social development in a still understudied primate species, these findings contribute to understanding the evolutionary roots of human maternal care and social development.


Mode of degradation of the chondroitin sulphate proteoglycan in rat costal cartilage.
1. Chondroitin sulphate was isolated from different regions of rat costal cartilage after extensive proteolysis of the tissues. The molecular weight, determined by gel chromatography, of the polysaccharide obtained from an actively growing region (lateral zone) near the osteochondral junction was higher than that of the polysaccharide isolated from the remaining portion of the costal cartilage (medial zone). 2. In both types of cartilage the molecular weight of chondroitin sulphate, labelled with [(35)S]sulphate, remained unchanged in vivo over a period of 10 days, approximately corresponding to the half-life of the chondroitin sulphate proteoglycan. The molecular-weight distribution of chondroitin [(35)S]sulphate, labelled in vivo or in vitro, was invariably identical with that of the bulk polysaccharide from the same tissue. It is concluded that the observed regional variations in molecular-weight distribution were established at the time of polysaccharide biosynthesis. 3. In tissue culture more than half of the (35)S-labelled polysaccharide-proteins of the two tissues was released into the medium within 10 days of incubation. The released materials were of smaller molecular size than were the corresponding native proteoglycans. In contrast, the molecular-weight distribution of the chondroitin [(35)S]sulphate (single polysaccharide chains) remained constant throughout the incubation period. 4. A portion (about 20%) of the total radioactive material released from (35)S-labelled cartilage in tissue culture was identified as inorganic [(35)S]sulphate. No corresponding decrease in the degree of sulphation of the labelled polysaccharide could be detected. These findings suggest that a limited fraction of the proteoglycan molecules had been extensively desulphated. 5. It is suggested that the initial phase of degradation involves proteolytic cleavage of the proteoglycan, but the constituent polysaccharide chains remain intact. The partially degraded proteoglycan may be eliminated from the cartilage by diffusion into the circulatory system. An additional degradative process, which may occur intracellularly, includes desulphation of the polysaccharide, probably in conjunction with a more extensive breakdown of the polymer.


Prehypertension, obesity, and risk of kidney disease: 20-year follow-up of the HUNT I study in Norway.
The combined effect of blood pressure (BP) and body weight on risk of kidney disease has not been previously studied. To improve risk stratification in prehypertensive individuals (ie, BP, 120 to 139/80 to 89 mm Hg), we examined the interaction between BP and body weight on the risk of end-stage renal disease or chronic kidney disease (CKD)-related death. Retrospective cohort study. 74,986 adults participating in the first Health Study in Nord-Trøndelag (88% participation rate) were linked to the Norwegian Renal Registry and Cause of Death Registry. BP and body weight were measured by using standard procedures, and other relevant covariates were obtained from an extensive questionnaire. Hazard ratios for treated end-stage renal disease and CKD-related death were calculated. Mean systolic BP and body mass index (BMI) were 136.8 +/- 23.3 (SD) mm Hg and 25.2 +/- 3.9 kg/m(2), whereas 12.9% had treated hypertension at baseline, respectively. During a median follow-up of 21 years (1,345,882 person-years), 507 men (1.4%) and 319 women (0.8%) initiated renal replacement therapy (n = 157) or died of CKD (n = 669). Multiadjusted risk of these kidney outcomes increased continuously with no lower threshold for BP. The risk associated with body weight started to increase from a BMI of 25.0 kg/m(2). In participants with BP less than 120/80 mm Hg, risk did not increase with increasing BMI. In prehypertensive participants, multivariate adjusted hazard ratios in the BMI categories 18.5 to 24.9, 25.0 to 29.9, 30.0 to 34.9, and 35.0 kg/m(2) or greater were 1.21 (95% confidence interval [CI], 0.67 to 2.17), 1.10 (95% CI, 59 to 2.00), 2.66 (95% CI, 1.28 to 5.53), and 5.94 (95% CI, 1.94 to 18.20) compared with BP less than 120/80 mm Hg and BMI of 18.5 to 24.9 kg/m(2), respectively (P = 0.02 for trend). Corresponding risks in hypertensive participants were 2.13 (95% CI, 1.23 to 3.70), 2.40 (95% CI, 1.40 to 4.15), 3.32 (95% CI, 1.89 to 5.81), and 5.53 (95% CI, 3.01 to 10.20), respectively (P < 0.001 for trend). Baseline creatinine measurements were not available; hence, a secondary analysis was performed that excluded all individuals who experienced outcomes in the 5 years after the study start. Participants with prehypertension are not at increased risk of serious kidney outcomes if BMI is less than 30.0 kg/m(2). However, the risk of kidney disease increases substantially if prehypertension is present in obese participants.


Projected Clinical Benefits of Implementation of SGLT-2 Inhibitors Among Medicare Beneficiaries Hospitalized for Heart Failure.
The sodium-glucose cotransporter-2 (SGLT-2) inhibitors form the latest pillar in the management of heart failure with reduced ejection fraction (HFrEF) and appear to be effective across a range of patient profiles. There is increasing interest in initiating SGLT-2 inhibitors during hospitalization, yet little is known about the putative benefits of this implementation strategy. We evaluated Medicare beneficiaries with HFrEF (≤ 40%) hospitalized at 228 sites in the Get With The Guidelines-Heart Failure (GWTG-HF) registry in 2016 who had linked claims data for ≥ 1 year postdischarge. We identified those eligible for dapagliflozin under the latest U.S. Food and Drug Administration label (excluding estimated glomerular filtration rates < 25 mL/min per 1.73 m2, dialysis and type 1 diabetes). We evaluated 1-year outcomes overall and among key subgroups (age ≥ 75 years, gender, race, hospital region, kidney function, diabetes status, triple therapy). We then projected the potential benefits of implementation of dapagliflozin based on the risk reductions observed in the Dapagliflozin and Prevention of Adverse Outcomes in Heart Failure (DAPA-HF) trial. Among 7523 patients hospitalized for HFrEF, 6576 (87%) would be candidates for dapagliflozin (mean age 79 ± 8 years, 39% women, 11% Black). Among eligible candidates, discharge use of β-blockers, ACEi/ARB, MRA, ARNI, and triple therapy (ACEi/ARB/ARNI+β-blocker+MRA) was recorded in 88%, 64%, 29%, 3%, and 20%, respectively. Among treatment-eligible patients, the 1-year incidence (95% CI) of mortality was 37% (36-38%) and of HF readmission was 33% (32-34%), and each exceeded 25% across all key subgroups. Among 1333 beneficiaries eligible for dapagliflozin who were already on triple therapy, the 1-year incidence of mortality was 26% (24%-29%) and the 1-year readmission due to HF was 30% (27%-32%). Applying the relative risk reductions observed in DAPA-HF, absolute risk reductions with complete implementation of dapagliflozin among treatment-eligible Medicare beneficiaries are projected to be 5% (1%-9%) for mortality and 9% (5%-12%) for HF readmission by 1 year. The projected number of Medicare beneficiaries who would need to be treated for 1 year to prevent 1 death is 19 (11-114), and 12 (8-21) would need to be treated to prevent 1 readmission due to HF. Medicare beneficiaries with HFrEF who are eligible for dapagliflozin after hospitalization due to HF, including those well-treated with other disease-modifying therapies, face high risks of mortality and HF readmission by 1 year. If the benefits of reductions in death and hospitalizations due to HF observed in clinical trials can be fully realized, the absolute benefits of implementation of SGLT-2 inhibitors among treatment-eligible candidates are anticipated to be substantial in this high-risk postdischarge setting.


Comparison of Antimicrobial Efficacy of Green Tea, Garlic with Lime, and Sodium Fluoride Mouth Rinses against Streptococcus mutans, Lactobacilli species, and Candida albicans in Children: A Randomized Double-blind Controlled Clinical Trial.
With greater awareness worldwide, the use of herbs and herbal products has increased to a large extent. To evaluate and compare the antimicrobial efficacy of green tea, garlic with lime, and 0.05% sodium fluoride (NaF) mouth rinses against Streptococcus mutans, Lactobacilli species, and Candida albicans. A total of 45 children aged 4 to 6 years with severe early childhood caries (S-ECC; based on decayed extracted filled [defs] score) were selected. Children were divided randomly into three equal groups and were asked to rinse with the prescribed mouth rinse once daily for 2 weeks after breakfast under supervision. A base-line and postrinsing nonstimulated whole salivary sample (2 mL) was collected and tested for the number of colony-forming units (CFUs). The data were statistically analyzed using Statistical Package for the Social Sciences (SPSS) version 16.0 software with one-way analysis of variance (ANOVA) and Tukey's post hoc test. A statistically significant fall in colony count was found with the three mouth rinses in S. mutans (p < 0.001, p < 0.001) and Lactobacilli spp. (p < 0.001, p < 0.001), but not against C. albicans (p = 0.264, p = 0.264). On comparison, no statistically significant difference was found against S. mutans (p = 1, p = 0.554, p = 0.572), lactobacilli spp. (p = 0.884, p = 0.999, p = 0.819), and C. albicans (p = 0.999, p = 0.958, p = 0.983). The findings of this study indicate that green tea and garlic with lime mouth rinse can be an economical alternative to NaF mouth rinse both for prevention and therapeutics. Thomas A, Thakur S, Habib R. Comparison of Antimicrobial Efficacy of Green Tea, Garlic with Lime, and Sodium Fluoride Mouth Rinses against Streptococcus mutans, Lactobacilli species, and Candida albicans in Children: A Randomized Double-blind Controlled Clinical Trial. Int J Clin Pediatr Dent 2017;10(3):234-239.


Dietary Antioxidant Insufficiency Is Associated With Increased Inflammatory Markers and Poorer Health-Related Quality of Life in Patients With Heart Failure.
Antioxidant insufficiency, elevated inflammatory markers, and poor health-related quality of life (HRQOL) are prevalent in patients with heart failure (HF). The objective of this study was to examine the associations among dietary antioxidant intake, inflammatory markers, and HRQOL in patients with HF. This was a secondary analysis of 265 patients with HF who completed a 4-day food diary. We assessed intake of 10 antioxidants: alpha carotene, beta carotene, beta cryptoxanthin, lutein, zeaxanthin, lycopene, vitamins C and E, zinc, and selenium. Antioxidant insufficiency was reflected by a measured level for each antioxidant that was below the estimate average requirement or lower than median for antioxidants without an estimate average requirement. Inflammatory markers including serum C-reactive protein, cytokines (interleukins 6 and 10), tumor necrosis factor-alpha, and soluble receptors (sTNFR1 and sTNFR2) were assessed with enzyme immunoassay. Health-related quality of life was measured using the Minnesota Living with Heart Failure at 12 months. Dietary antioxidant insufficiency predicted C-reactive protein (β = 0.135, P = .032) and interleukin 10 (β = -.155, P = .027). Patients with higher antioxidant insufficiency had higher C-reactive protein and lower interleukin 10. Both antioxidant insufficiency (β = 0.13, P = .049) and higher C-reactive protein (β = 0.16, P = .019) were independently associated with poorer HRQOL while adjusting for covariates. Dietary antioxidant insufficiency was associated with increased markers of inflammation and poorer HRQOL. Improvement of diet quality among patients with HF may be a fruitful area of research for enhancing HRQOL.


Electrically switchable liquid crystal Fresnel lens using UV-modified alignment film.
A simple method to make a switchable liquid crystal (LC) Fresnel lens with high diffraction efficiency and a low driving voltage was proposed based on the photo-induced surface modification of the vertical alignment layer. UV illumination alters the pretilt angle of alignment layers, a Fresnel zone-distribution hybrid alignment in the homeotropic LC cell can be straightforwardly achieved through UV exposure, yielding a concentric structure of the Fresnel phase LC lens. A remarkable diffraction efficiency of ~31.4%, close to the measured diffraction efficiency of the used Fresnel-zone-plate mask of 32%, was detected using a linearly polarized incident beam.


Discriminative Accuracy of Chronic Obstructive Pulmonary Disease Screening Instruments in 3 Low- and Middle-Income Country Settings.
Most of the global morbidity and mortality in chronic obstructive pulmonary disease (COPD) occurs in low- and middle-income countries (LMICs), with significant economic effects. To assess the discriminative accuracy of 3 instruments using questionnaires and peak expiratory flow (PEF) to screen for COPD in 3 LMIC settings. A cross-sectional analysis of discriminative accuracy, conducted between January 2018 and March 2020 in semiurban Bhaktapur, Nepal; urban Lima, Peru; and rural Nakaseke, Uganda, using a random age- and sex-stratified sample of the population 40 years or older. Three screening tools, the COPD Assessment in Primary Care to Identify Undiagnosed Respiratory Disease and Exacerbation Risk (CAPTURE; range, 0-6; high risk indicated by a score of 5 or more or score 2-5 with low PEF [<250 L/min for females and <350 L/min for males]), the COPD in LMICs Assessment questionnaire (COLA-6; range, 0-5; high risk indicated by a score of 4 or more), and the Lung Function Questionnaire (LFQ; range, 0-25; high risk indicated by a score of 18 or less) were assessed against a reference standard diagnosis of COPD using quality-assured postbronchodilator spirometry. CAPTURE and COLA-6 include a measure of PEF. The primary outcome was discriminative accuracy of the tools in identifying COPD as measured by area under receiver operating characteristic curves (AUCs) with 95% CIs. Secondary outcomes included sensitivity, specificity, positive predictive value, and negative predictive value. Among 10 709 adults who consented to participate in the study (mean age, 56.3 years (SD, 11.7); 50% female), 35% had ever smoked, and 30% were currently exposed to biomass smoke. The unweighted prevalence of COPD at the 3 sites was 18.2% (642/3534 participants) in Nepal, 2.7% (97/3550) in Peru, and 7.4% (264/3580) in Uganda. Among 1000 COPD cases, 49.3% had clinically important disease (Global Initiative for Chronic Obstructive Lung Disease classification B-D), 16.4% had severe or very severe airflow obstruction (forced expiratory volume in 1 second <50% predicted), and 95.3% of cases were previously undiagnosed. The AUC for the screening instruments ranged from 0.717 (95% CI, 0.677-0.774) for LFQ in Peru to 0.791 (95% CI, 0.770-0.809) for COLA-6 in Nepal. The sensitivity ranged from 34.8% (95% CI, 25.3%-45.2%) for COLA-6 in Nepal to 64.2% (95% CI, 60.3%-67.9%) for CAPTURE in Nepal. The mean time to administer the instruments was 7.6 minutes (SD 1.11), and data completeness was 99.5%. This study demonstrated that screening instruments for COPD were feasible to administer in 3 low- and middle-income settings. Further research is needed to assess instrument performance in other low- and middle-income settings and to determine whether implementation is associated with improved clinical outcomes.


Effects of zinc on tissue uptake and toxicity of lead in Sprague Dawley rat.
Lead (Pb) exposure occurs together with other metals including zinc (Zn). This study investigated the impact of Zn on Pb tissue accumulation and Pb-induced toxicities. Animals (n=6 rats per group) were exposed to lead acetate (PbAc) or a combination of PbAc and zinc acetate (ZnAc) under the following groups: control (deionized water), low PbAc [12 mg/kg PbAc (3 mg PbAc/rat/day)], low PbAc-ZnAc [12 mg/kg PbAc (3 mg PbAc/rat/day) + 0.2 mg ZnAc/rat/48 hr], high PbAc [120 mg/kg (30 mg PbAc/rat/day)], and high PbAc-ZnAc [120 mg/kg (30 mg PbAc/rat/day) + 1 mg ZnAc/rat/48 hr] for 8 weeks. A significant reduction in body weight gain was observed in the high PbAc group relative to the control group. Muscles and testes both had reduced and increased Pb uptake in low PbAc-ZnAc and high PbAc-ZnAc groups compared to PbAc only groups, respectively. Bone Pb levels in the high PbAc-ZnAc group were lower than the high PbAc group. Zinc co-administration attenuated Pb-induced inhibition of delta aminolaevulinic acid dehydratase enzyme and enhanced catalase enzyme activity at a high level of exposure. Moreover, ZnAc seems to have minimized the effects of Pb-induced mRNA dysregulation in antioxidant and antiapoptotic enzymes encoding genes. Heme oxygenase-1 was downregulated in the kidney and brain in the low PbAc group. Liver glutathione peroxidase and thioredoxin reductase-1 were downregulated in the high PbAc group. These findings suggest that zinc co-administration with lead may partially mitigate against Pb-induced toxicities.


Bovine lactoferrin prevents invasive fungal infections in very low birth weight infants: a randomized controlled trial.
Lactoferrin is a mammalian milk glycoprotein involved in innate immunity. Recent data show that bovine lactoferrin (bLF) prevents late-onset sepsis in preterm very low birth weight (VLBW) neonates. This is a secondary analysis of data from a multicenter randomized controlled trial where preterm VLBW neonates randomly received bLF (100 mg/day; group A1), bLF + Lactobacillus rhamnosus GG (10(6) colony-forming units per day; group A2), or placebo (group B) for 6 weeks. Here we analyze the incidence rates of fungal colonization, invasive fungal infection (IFI), and rate of progression from colonization to infection in all groups. This study included 472 neonates whose clinical, nutritional, and demographical characteristics were similar. Overall, the incidence of fungal colonization was comparable (17.6%, 16.6%, and 18.5% in A1, A2, and B, respectively; P = .89 [A1] and .77 [A2]). In contrast, IFIs were significantly decreased in A1 and A2 (0.7% and 2.0%, respectively) compared with B (7.7%; P = .002 [A1] and .02 [A2]), and this was significantly true both in <1000 g (0.9% [A1] and 5.6% [A2], vs 15.0%) and in 1001 to 1500 g infants (0% and 0% vs 3.7%). The progression rate colonization-infection was significantly lower in the bLF groups: 3.7% (A1) and 12% (A2), vs 41.9%; P < .001 (A1) and P = .02 (A2). No IFI-attributable deaths occurred in the treatment groups, versus 2 in placebo. No adverse effects or intolerances occurred. Prophylactic oral administration of bLF reduces the incidence of IFI in preterm VLBW neonates. No effect is seen on colonization. The protective effect on IFI is likely due to limitation of ability of fungal colonies to progress toward invasion and systemic disease in colonized infants.


Vitamin D receptor gene polymorphism and serum vitamin D level as risk factors for acquiring Type II diabetes mellitus.
The studies on the relationship between type 2 diabetes mellitus (T2DM) risk and vitamin D receptor (VDR) gene polymorphisms are still inconclusive. Therefore, the objective of the study was to assess possible risks of acquiring T2DM due to polymorphisms in the VDR gene or abnormal serum levels of VD. 362 participants (181 T2DM patients and 181 healthy controls) from the Diabetic Center, Sulaimaniyah/Iraq, from December 2020 to May 2021 were volumtarily enrolled in the study. For each participant, HbA1c, fasting blood sugar (FBS), serum cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG), markers of calcium homeostasis, alkaline phosphatase (ALP), phosphorus, VD and insulin were measured. In addition, FokI, TaqI, ApaI, and BsmI genotypes were also performed using Polymerase Chain Reaction (PCR). The results showed that VD level was significantly lower in T2DM compared to the control group. While, HbA1c was significantly higher in T2DM than in the control group. In contrast to AA (P=0.034) and CC (P=0.011) genotype of ApaI (rs7975232) gene polymorphism, which were dominant among the control group, AC-genotype was significantly (P=0.0001) dominat among T2DM group. Meanwhile, TT-genotype of TaqI (rs731236) was significantly (P=0.05) dominant among control group. While there were not any significant differences between other genotypes among T2DM and control groups. In conclusion, low VD-level is a possible risk factor for developing T2DM, and an association was found, especially between ApaI genotypes and T2DM.


Somatic or sympathetic block for reflex sympathetic dystrophy. Which is indicated?
Patients with reflex sympathetic dystrophy (complex regional pain syndromes) are often referred to pain medicine physicians for assistance in providing pain control during rehabilitation of their painful upper extremity. When deciding which pain control technique to use in an individual patient, physicians must consider both somatic and sympathetic blocks. Each of these nerve blocks has advantages and disadvantages that may be tailored to an individual patient's pain state and rehabilitation program to optimize recovery.


Food-induced anaphylaxis.
Food-induced anaphylaxis is the leading single cause of anaphylaxis treated in emergency departments and increasing in prevalence. Food allergy is an increasing problem in westernized countries around the world, with a cumulative prevalence of 3-6%. Peanut, tree nuts, and shellfish are the most commonly implicated foods in anaphylaxis, although milk is a common trigger in children. Asthmatics, adolescents, and those with a prior reaction are at increased risk for more severe reactions. Most first reactions and reactions in children most commonly occur at home, whereas most subsequent reactions and reactions in adults occur outside home. Studies on schools have identified inadequate management plans and symptom recognition whereas those on restaurants report lack of prior notification by allergic individuals and lack in staff education. Epinephrine, although underutilized is the drug of choice with multiple doses needed in up to one-fifth of reactions. Diagnosis is currently based on convincing history and allergy testing supported by elevated serum tryptase, if available. Long-term management includes strict avoidance and emergency action plan. With a growing population of food-allergic children and adults, markers to predict which individuals are at increased risk for anaphylaxis as well as new therapies are vigorously sought.


Effects of detraining on enzymes of energy metabolism in individual human muscle fibers.
Muscle biopsies were obtained from three cyclists and four runners at the end of 10-24 mo of intensive training and after intervals of detraining up to 12 wk. Control samples came from four untrained persons and four former athletes. Macro mixed fiber samples were assayed for lactate dehydrogenase, adenylate kinase, glycogen phosphorylase, citrate synthase, malate dehydrogenase, beta-hydroxyacyl-CoA dehydrogenase, succinate dehydrogenase, beta-hydroxybutyrate dehydrogenase, creatine kinase, hexokinase, 1-phosphofructokinase, fructosebisphosphatase, protein, and total creatine. In the case of three trained persons and two controls, the first six of the enzymes were also measured in individual fibers. Before detraining, enzymes of oxidative metabolism were substantially higher than in controls, and differences in levels between type I and type II fibers were smaller. During detraining, oxidative enzymes were decreased in both fiber types but the type II fibers did not fall to control levels even after 12 wk. Phosphorylase increased with detraining in both fiber types. The same is true for lactate dehydrogenase and adenylate kinase, except in the case of the type I fibers of one individual. Among the other six enzymes (measured in mixed fiber samples), only hexokinase was consistently affected (decreased) by detraining.


Radioimmunodetection: technical problems and methods of improvement.
Radioimmunodetection (RAID) is a technique which uses radiolabelled antibodies to visualize tumours, taking advantage of antigens preferentially expressed by malignant tissue. Gamma radiation emitted by radioisotopes can be detected using an external gamma camera (RAID), or intraoperatively with a hand-held Geiger counter (radioimmunoguided surgery, RIGS). RAID has significant inherent problems. Many have been overcome as a result of nearly 50 years of research, and others still remain as obstacles precluding the routine use of the technique. This article summarizes the technical limitations of RAID and outlines the relative successes of the methods evolved to overcome them.


Systematic review of room temperature stability of key beta-lactam antibiotics for extended infusions in inpatient settings.
Extended infusion (EI) of beta-lactam antibiotics may offer clinical benefits aligned with improved probability of target attainment for critical pharmacokinetic/pharmacodynamic parameters that correlate with efficacy. There is much research interest in prolonged and continuous infusions (collectively, extended infusions) of beta-lactams to improve patient outcomes, particularly in critically ill patients in intensive care. While definitive clinical trial data demonstrating beneficial outcomes is awaited, there has been limited focus on the stability of the agents given by EI, which may be an equally critical parameter. EI may allow for savings in nursing time due to reduced need for drug reconstitution. We set out to examine the data for stability for EI at room temperature, consistent with the requirements of 'A Standard Protocol for Deriving and Assessment of Stability- Part 1 Aseptic Preparation (Small Molecules)', which allows a 5% loss of active pharmaceutical ingredient (API) applicable for those territories that use the British Pharmacopoeia also for a 10% loss applicable in much of rest of the world. Searches using preferred reporting items for systematic reviews and meta-analyses (PRISMA) principles for stability data on freshly prepared beta-lactam antimicrobials for extended administration at room temperature (at or above 23°C) were conducted in November 2021 and updated in December 2022. We found data to support the extension of the shelf life of 12 key beta-lactam antibiotics once reconstituted (aztreonam, amoxicillin, benzylpenicillin, flucloxacillin, piperacillin/tazobactam, cefazolin, cefmetazole, ceftaroline, ceftazidime, ceftriaxone, imipenem and meropenem) compliant with the NHS protocol, and data for five other agents (ticarcillin, cefepime, cefiderocol, cefoxitin and doripenem) which would be acceptable in regions outside the UK beyond that listed in the Summary of Product Characteristics.This review has not been registered under PROSPERO.


[Effect of Danshen-containing serum on expression of SuFu and DYRK2 in HSCs].
To observe the effects of Danshen-containing serum on SuFu and DYRK2 expression in the HSCs stimulated by leptin. SD rats (n = 60) were used to make danshen-containing serum by gastric perfusion for ten days with Danshen water decoction, normal saline and colchicine. The HSCs that were cultured in vitro would be stimulated for 24 hours by leptin (100 μg x L(-1)) except blank control group, after being intervened, the drug serum in each group would be cultured at 37 degrees C in 5% incubator. The cells would be collected after 24 hours, then the effects of danshen-containing serum on the proliferation of HSCs were detected by MTT, the expression of SuFu mRNA and DYRK2 mRNA were detected by RT-PCR, the expression of SuFu and DYRK2 proteins were tested by Western blot. Compared with blank control group, the expression of DYRK2 mRNA and DYRK2 proteins were enhanced obviously after stimulated the HSCs of rats by leptin (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, after cyclopamine group (Hh pathway inhibitor), Danshen-containing serum and colchicine were interfered, the expression of DYRK2 mRNA and DYRK2 proteins were decreased clearly (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were increased significantly (P < 0.01 or P < 0.05). Compared with model group, adding purmorphamine (Hh pathway agonist) to model group and making it activate could increase the expression of DYRK2 mRNA and DYRK2 proteins, but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, using the Danshen-containing serum to interfere the purmorphamine group could make the expression of DYRK2 mRNA and DYRK2 proteins decrease and the expression of SuFu mRNA and SuFu proteins increase significantly (P < 0.01). Danshen-containing serum would inhibition the activation and increment of HSCs by interfering the expression of SuFu and DYRK2 which were induced by leptin.


Ionizing radiation and genetic risks. XI. The doubling dose estimates from the mid-1950s to the present and the conceptual change to the use of human data on spontaneous mutation rates and mouse data on induced mutation rates for doubling dose calculations.
This paper provides an overview of the concept of doubling dose, changes in the database employed for calculating it over the past 30 years and recent advances in this area. The doubling dose is estimated as a ratio of the average rates of spontaneous and induced mutations in a defined set of genes. The reciprocal of the doubling dose is the relative mutation risk per unit dose and is one of the quantities used in estimating genetic risks of radiation exposures. Most of the doubling dose estimates used thus far have been based on mouse data on spontaneous and induced rates of mutations. Initially restricted to mutations in defined genes (with particular focus on the seven genes at which induced recessive mutations were studied in different laboratories), the doubling dose concept was subsequently expanded to include other endpoints of genetic damage. At least during the past 20 years, the magnitude of the doubling dose has remained unchanged at approximately 1 Gy for chronic low LET radiation exposures. One of the assumptions underlying the use of the doubling dose based on mouse data for predicting genetic risks in humans, namely, that the spontaneous rates of mutations in mouse and human genes are similar, is incorrect; this is because of the fact that, unlike in the mouse, the mutation rate in humans differs between the two sexes (being higher in males than in females) and increases with paternal age. Further, an additional source of uncertainty in spontaneous mutation rate estimates in mice has been uncovered. This is related to the non-inclusion of mutations which arise as germinal mosaics and which result in clusters of identical mutations in the following generation. In view of these reasons, it is suggested that a prudent way forward is to revert to the use of human data on spontaneous mutation rates and mouse data on induced mutation rates for doubling dose calculations as was first done in the 1972 BEIR report of the US National Academy of Sciences. The advantages of this procedure are the following: (i) estimates of spontaneous mutation rates in humans, which are usually presented as sex-averaged rates, automatically include sex differences and paternal age-effects; (ii) since human geneticists count all mutations that arise anew irrespective of whether they are part of a cluster or not, had clusters occurred, they would have been included in mutation rate calculations and (iii) one stays close to the aim of risk estimation, namely, estimation of the risk of genetic diseases in humans. On the basis of detailed analyses of the pertinent data, it is now estimated that the average spontaneous mutation rate of human genes (n=135 genes) is: (2.95+/-0.64)x10(-6) per gene and the average induced mutation rate of mouse genes (n=34) is: (0.36+/-0.10)x10(-5) per gene per Gy for chronic low LET radiation. The resultant doubling dose is (0.82+/-0.29) Gy. The standard error of the doubling dose estimate incorporates sampling variability across loci for estimates of spontaneous and induced mutation rates as well as variability in induced mutation rates in individual mouse experiments on radiation-induced mutations. We suggest the use of a rounded doubling dose value of 1 Gy for estimating genetic risks of radiation. Although this value is the same as that used previously, its conceptual basis is different and the present estimate is based on more extensive data than has so far been the case.


Comparison of N-acyl phosphatidylethanolamines with different N-acyl groups as activators of glucocerebrosidase in various forms of Gaucher's disease.
The acidic phospholipid requirement of the predominant particulate beta-glucosidase of mammalian spleen and liver was investigated using a series of N-acyl derivatives of dioleoyl phosphatidylethanolamine (PE). The PE, a neutral phospholipid, was converted to an acidic lipid, (N-acyl)-phosphatidylethanolamine (NAPE) by acylation of the amino group with different fatty acyl chains. Lysosomal beta-glucosidases from rat liver and spleens of controls and patients with various types of Gaucher's disease were solubilized and delipidated by extraction with sodium cholate and 1-butanol. All members of the NAPE series tested were effective activators of the delipidated rat liver beta-glucosidase, and the stimulatory power of the NAPE family increased with increasing chain length of the fatty acid substitution. In contrast, dioleoyl-PE had no effect on beta-glucosidase activity. A heat-stable factor (HSF) purified from the spleen of a patient with Gaucher's disease significantly increased the sensitivity of the rat liver beta-glucosidase to all of the NAPE derivatives. The maximum stimulation achieved in the presence of HSF was independent of N-acyl chain length. Compared to the potent activator, phosphatidylserine (PS), (N-acetyl)-PE and (N-linoleoyl)-PE were half as effective as activators of beta-glucosidase from control human spleen. PS stimulated the beta-glucosidase of type 1 nonneurologic Gaucher's disease, but none of the NAPE compounds activated it. Neither PS nor any of the (N-acyl)-PE compounds could activate a delipidated preparation of beta-glucosidase obtained from the spleen of a neurologic case. These results indicate that although the presence of a net negative charge on a phospholipid confers upon it an ability to reconstitute beta-glucosidase activity to the normal, nonmutant enzyme, it is insufficient to permit differentiation of the various types of Gaucher's disease.


The Role of Experience and Gender in Founders' Business Planning Activities: A Meta Analysis.
The question of why entrepreneurs undertake business planning activities differently, ranging from planning "in the head" to generating formal written documents, is still impenetrable. Aggregating data on 11,064 observations from 32 independent data set, this study meta-analyzed how business experience and gender influence entrepreneurs' disposition to business planning behaviors. Surprisingly, contradictory to some extant views that entrepreneurs without prior experience are more likely to make business plans, we found that both managerial experience and entrepreneurial experience positively influence entrepreneurs' subsequent business planning behaviors. Drawing insight from the effectuation and institutional perspectives, this study showed that, rather than entrepreneurial experience, managerial experience motivates entrepreneurs to generate formal business plans. For entrepreneurs who create formal business plans, both entrepreneurial experience and managerial experience enhance their business planning sophistication. In addition, we examined the moderating effects of gender on the relationship between business experience and business planning. The results suggested that female entrepreneurs with entrepreneurial experience are more likely to undertake business planning behaviors and create formal business plans than their male counterparts.


An Update on Biochemical and Genomic Markers for Prostate Cancer.
Detecting prostate cancer, developing therapeutic plans after negative biopsies, and prognosis-based patient counseling can be challenging for many urologists dealing with prostate cancer-specific antigens. New Biomarkers advances made improvement for prediction of responses to therapeutic option and can tell us about survival and recurrence. In this review, we have assessed current and upcoming biomarkers that are opening a new era in diagnosing the disease. We conducted a comprehensive literature review of studies describing prostate cancer biomarkers. Two independent investigators searched PubMed, Embase, Web of Science, and Cochrane Databases to identify biomarkers in prostate cancer conducted a literature review. Recently, combining prostate cancer-specific biomarkers into a single test has gained increasing attention, especially since the introduction of genomic and molecular tools. The development of the Prostate Health Index (PHI), SelectMDx, and Confirm MDx have shown promising results for prostate cancer detection, in addition to risk stratification and biopsy avoidance. Despite major improvements and innovations in prostate cancer biomarkers, application in current clinical practice is limited. However, these biomarkers have an important role in determining risk, preventing unnecessary prostate biopsies, and predicting prognoses. Additional confirmatory studies will be needed to fully understand the impact of prostate cancer-specific biomarkers.


Hypoxia and extracellular matrix proteins influence angiogenesis and lymphangiogenesis in mouse embryoid bodies.
Regulatory mechanisms for angiogenesis are relatively well established compared to lymphangiogenesis. Few studies have shown that a combination of vascular endothelial growth factor VEGF-A/C with hypoxia or collagen matrix promotes lymphatic structures along with blood vessel development in mouse embryoid bodies (EB). In this study we tested the hypothesis that while hypoxia combined with prolonged VEGF-A/C treatment would induce early lymphangiogenesis in addition to angiogenesis in mouse EBs, under similar conditions specific extracellular matrix (ECM) proteins would promote lymphatic vessel-like structures over angiogenesis. EBs were subjected to four conditions and were maintained under normoxia and hypoxia (21% and 2.6% O(2), respectively) with or without VEGF-A/C. Microarray analyses of normoxic and hypoxic EBs, and immunofluorescence data showed very low expression of early lymphatic endothelial cell (LEC) markers, lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), and prospero-related homeobox 1 (Prox1) at early time points. Double immunofluorescence using MECA-32 and Prox1/LYVE1 demonstrated that combined hypoxia and VEGF-A/C treatment promoted formation of blood vessel-like structures, whereas only Prox1(+)/LYVE1(+) LECs were detected in EBs at E22.5. Furthermore, EBs were grown on laminin or collagen-I coated plates and were subjected to the four treatments as described above. Results revealed that LECs in EBs at E36.5 attached better to collagen-I, resulting in an organized network of lymphatic vessel-like structures as compared to EBs grown on laminin. However, blood vessel-like structures were less favored under these same conditions. Collectively, our data demonstrate that hypoxia combined with growth factors promotes angiogenesis, whereas combination of these conditions with specific ECM proteins favors lymphangiogenesis processes in mouse EBs.


Dense junctures of ethical concern.
A collection of stories by bioethicists writing about their own illnesses displays the importance of microethics. From this perspective, ethics happens not in the application of principles to specific decisions, but rather in the moment-to-moment flow of clinical interaction, as healthcare workers and patients make decisions, especially in their use of language. Microethical issues that are common to multiple stories are described as dense junctures of ethical concern. Three junctures are discussed in detail: conflicts between medical and patient rationalities, issues of how clinicians represent patients, and practices of truth telling. These junctures are illustrated with examples from the bioethicists' personal stories of treatment.


Targeting ganglioside epitope 3G11 on the surface of CD4+ T cells suppresses EAE by altering the Treg/Th17 cell balance.
Loss of expression of the 3G11 epitope, present on disialoceramide that is predominantly found on CD4(+) T cells, has been associated with a regulatory T cell (Treg) phenotype and tolerance induction in experimental autoimmune encephalomyelitis (EAE). Here we report that treatment with anti-3G11 mAb shifts the immune response from pro-inflammatory to tolerogenic and suppresses both chronic-progressive and relapsing-remitting EAE. This therapeutic effect can be achieved at different stages of EAE. Treatment with anti-3G11 mAb increased the proportion of Foxp3(+)CD25(+)CD4(+) Tregs and IL-10 production while inhibiting production of pro-inflammatory cytokines and responsiveness to IL-2 and decreasing the proportion of T(h)17 cells. The effect of anti-3G11 mAb was diminished in IL-10(-/-) mice, indicating that this cytokine mediates some of its effects. As 3G11 belongs to the ganglioside family, which is expressed on the surface of both murine and human CD4(+) T cells, targeting this class of molecules may provide a novel approach for treating autoimmune diseases.


Uptake of oxidized LDL by macrophages results in partial lysosomal enzyme inactivation and relocation.
The cytotoxicity of oxidized LDL (oxLDL) to several types of artery wall cells might contribute to atherosclerosis by causing cell death, presumably by both apoptosis and necrosis. After its uptake into macrophage lysosomes by receptor-mediated endocytosis, oxLDL is poorly degraded, resulting in ceroid-containing foam cells. We studied the influence ofoxLDL on lysosomal enzyme activity and, in particular, on lysosomal membrane stability and the modulation of these cellular characteristics by HDL and vitamin E (vit-E). Unexposed cells and cells exposed to acetylated LDL (AcLDL) were used as controls. The lysosomal marker enzymes cathepsin L and N-acetyl-beta-glucosaminidase (NAbetaGase) were biochemically assayed in J-774 cells after fractionation. Lysosomal integrity in living cells was assayed by the acridine orange (AO) relocation test. Cathepsin D was immunocytochemically demonstrated in J-774 cells and human monocyte-derived macrophages. We found that the total activities of NAbetaGase and cathepsin L were significantly decreased, whereas their relative cytosolic activities were enhanced, after oxLDL exposure. Labilization of the lysosomal membranes was further proven by decreased lysosomal AO uptake and relocation to the cytosol of cathepsin D, as estimated by light and electron microscopic immunocytochemistry. HDL and vit-E diminished the cytotoxicity of oxLDL by decreasing the lysosomal damage. The results indicate that endocytosed oxLDL not only partially inactivates lysosomal enzymes but also destabilizes the acidic vacuolar compartment, causing relocation of lysosomal enzymes to the cytosol. Exposure to AcLDL resulted in its uptake with enlargement of the lysosomal apparatus, but the stability of the lysosomal membranes was not changed.


Modelling of the break force of tablets under diametrical compression.
A numerical method to predict the break force of curved faced tablets is proposed. The constitutive model and the failure criteria necessary to obtain predictions consistent with experimental data are identified. A modified Drucker-Prager cap model together with a maximum principal stress based failure criteria was found sufficient to predict the break force of tablets under diametrical compression loading. The conditions for the validity of the method were identified with reference to practical tablet shapes and failure patterns. Under these conditions the numerical procedures can be used as a practical tool to predict tablet breakage as an alternative to the empirical methods currently used in pharmaceutical product design and process development.


IgG and IgM cooperate in coating of intestinal bacteria in IgA deficiency.
Immunoglobulin A (IgA) is acknowledged to play a role in the defence of the mucosal barrier by coating microorganisms. Surprisingly, IgA-deficient humans exhibit few infection-related complications, raising the question if the more specific IgG may help IgM in compensating for the lack of IgA. Here we employ a cohort of IgA-deficient humans, each paired with IgA-sufficient household members, to investigate multi-Ig bacterial coating. In IgA-deficient humans, IgM alone, and together with IgG, recapitulate coating of most bacterial families, despite an overall 3.6-fold lower Ig-coating. Bacterial IgG coating is dominated by IgG1 and IgG4. Single-IgG2 bacterial coating is sparse and linked to enhanced Escherichia coli load and TNF-α. Although single-IgG2 coating is 1.6-fold more prevalent in IgA deficiency than in healthy controls, it is 2-fold less prevalent than in inflammatory bowel disease. Altogether we demonstrate that IgG assists IgM in coating of most bacterial families in the absence of IgA and identify single-IgG2 bacterial coating as an inflammatory marker.


Removal of Mannose-Ending Glycan at Asn2118 Abrogates FVIII Presentation by Human Monocyte-Derived Dendritic Cells.
The development of an immune response against therapeutic factor VIII is the major complication in hemophilia A patients. Oligomannose carbohydrates at N239 and/or N2118 on factor VIII allow its binding to the macrophage mannose receptor expressed on human dendritic cells, thereby leading to factor VIII endocytosis and presentation to CD4+ T lymphocytes. Here, we investigated whether altering the interaction of factor VIII with mannose-sensitive receptors on antigen-presenting cells may be a strategy to reduce factor VIII immunogenicity. Gene transfer experiments in factor VIII-deficient mice indicated that N239Q and/or N2118Q factor VIII mutants have similar specific activities as compared to non-mutated factor VIII; N239Q/N2118Q mutant corrected blood loss upon tail clip. Production of the corresponding recombinant FVIII mutants or light chains indicated that removal of the N-linked glycosylation site at N2118 is sufficient to abrogate in vitro the activation of FVIII-specific CD4+ T cells by human monocyte-derived dendritic cells. However, removal of mannose-ending glycans at N2118 did not alter factor VIII endocytosis and presentation to CD4+ T cells by mouse antigen-presenting cells. In agreement with this, the N2118Q mutation did not reduce factor VIII immunogenicity in factor VIII-deficient mice. Our results highlight differences in the endocytic pathways between human and mouse dendritic cell subsets, and dissimilarities in tissue distribution and function of endocytic receptors such as CD206 in both species. Further investigations in preclinical models of hemophilia A closer to humans are needed to decipher the exact role of mannose-ending glycans in factor VIII immunogenicity.


Unraveling self-assembly pathways of the 468-kDa proteolytic machine TET2.
The spontaneous formation of biological higher-order structures from smaller building blocks, called self-assembly, is a fundamental attribute of life. Although the protein self-assembly is a time-dependent process that occurs at the molecular level, its current understanding originates either from static structures of trapped intermediates or from modeling. Nuclear magnetic resonance (NMR) spectroscopy has the unique ability to monitor structural changes in real time; however, its size limitation and time-resolution constraints remain a challenge when studying the self-assembly of large biological particles. We report the application of methyl-specific isotopic labeling combined with relaxation-optimized NMR spectroscopy to overcome both size- and time-scale limitations. We report for the first time the self-assembly process of a half-megadalton protein complex that was monitored at the structural level, including the characterization of intermediate states, using a mutagenesis-free strategy. NMR was used to obtain individual kinetics data on the different transient intermediates and the formation of final native particle. In addition, complementary time-resolved electron microscopy and native mass spectrometry were used to characterize the low-resolution structures of oligomerization intermediates.


Epidemiology of gonococcal and chlamydial infections in Harrow and Brent.
To describe the epidemiology and associated clinical features of gonorrhoea and chlamydial infection and to develop a profile of sexually transmitted diseases (STDs) in an outer London health district. Hospital-based retrospective study. Genitourinary medicine clinic, Northwick Park Hospital, Harrow and Brent Health Authority. 70 male and female individuals with gonorrhoea and 129 with chlamydial infection, diagnosed consecutively over 28 months in 1992-94. More men than women had gonorrhoea (43 versus 27) but more women than men had chlamydial infection (84 versus 45), p < 0.001. There was a clear tendency for cases with either infection to locate along major road and rail transport routes. Foci of gonococcal infection were concentrated mainly in the densely populated areas, whereas chlamydial cases were more evenly spread. There was no significant effect of gender or type of STD on the odds ratio for residence in Harrow, single marital status or attendance for test of cure. However, the odds ratios for women having sexual intercourse with a regular partner only or previous STD were 5 (95% CI 2.4 to 10.2), p < 0.001 and 0.3 (95% CI 0.18 to 0.69), p = 0.002 times the odds for men, respectively. The odds ratios for patients with gonococcal infection being employed or having sex with a regular partner only were 0.5 (95% CI 0.27 to 0.98), p = 0.04 and 0.30 (95% CI 0.15 to 0.60), p < 0.001 times the odds for patients with chlamydial infection, respectively. Of the women with gonorrhoea and previous pregnancy, 68% gave a history of abortion compared with 44% of those with chlamydial infection (p = 0.03). The identification of foci gonococcal and chlamydial infection and apparent location of these infections along the major transport routes in our health district require further study. That chlamydial infection, unlike gonorrhoea, is evenly distributed irrespective of population concentration and deprivation, suggests urgent need for a comprehensive local effort to control both STDs.