Differential effects of dietary whey, casein and soya on colonic DNA damage and large bowel SCFA in rats fed diets low and high in resistant starch.
Feeding higher levels of dietary animal protein (as casein or red meat) increases colonic DNA damage and thins the colonic mucus barrier in rats. Feeding resistant starch (RS) reverses these changes and increases large bowel SCFA. The present study examined whether high dietary dairy (casein or whey) or plant (soya) proteins had similar adverse effects and whether dietary RS was protective. Adult male rats were fed diets containing 15 or 25 % casein, whey or soya protein with or without 48 % high amylose starch (as a source of RS) for 4 weeks. DNA damage was measured in isolated colonocytes using the comet assay. Higher dietary casein and soya (but not whey) increased colonocyte DNA damage. DNA damage was highest with soya when fed at 15 or 25 % protein without RS. Dietary RS attenuated protein-induced colonocyte DNA damage in all groups but it remained significantly higher in rats fed 25 % soya compared with those fed 15 % protein. Dietary protein level did not affect colonic mucus thickness overall but the barrier was thinner in rats fed high dietary casein. This effect was reversed by feeding RS. Caecal total SCFA and butyrate pools were higher in rats fed RS compared with digestible starch. Caecal and faecal SCFA were unrelated to genetic damage but correlated with mucus thickness. The present data confirm that higher dietary protein affected colonocyte DNA and colonic mucus thickness adversely but that proteins differ in their effects on these indices of colon health. The data show also that these changes were reversed by RS.


Evaluation of growth monitoring programme in children in Kinshasa.
A high dropout rate was noted at the under five clinic of the University of Kinshasa Teaching Hospital. A preliminary study carried out in March 1984 on 197 children indicated that none of them had completed the five year period of the growth monitoring programme. Five hundred mothers of children attending the under five year clinic were then randomly selected from the 1224 who registered between September 1983 and August 1984. They were interviewed at home during a six week period. This paper confirms the dropout problem. It discusses whether mothers should abandon the visits early. It concludes that health workers should be trained to advise mothers to bring their children to the programme up to five years though the attendance frequency may be reduced from the second year onwards.


Quality of osteoarthritis care for community-dwelling older adults.
To describe the quality of osteoarthritis care provided to community-dwelling elderly patients and to characterize arthritis-related function in these patients. Two medical groups in the western United States participated in a practice-redesign intervention targeted at falls and mobility disorders, incontinence, and cognitive impairment, but not osteoarthritis. From 339 individuals reporting a diagnosis of arthritis, we collected information on demographics, functional status, and quality of care via patient interviews and mailed questionnaires. Eight quality indicators measured osteoarthritis care: 4 indicators measuring the provision of effective osteoarthritis care (osteoarthritis treatment indicators) and 4 measuring the provision of safe osteoarthritis care (medication safety indicators). The mean +/- SD Short Form 12 physical component summary score was 35 +/- 11 points, indicating a physically frail population. The overall indicator pass rate was 57.0% (95% confidence interval [95% CI] 53.9-60.2). Pass rates were higher for indicators of osteoarthritis treatment (63.5%, 95% CI 59.8-67.2) than for indicators of medication safety (43.8%, 95% CI 38.2-49.4). Patients with hip or knee pain had mean +/- SD Western Ontario and McMaster Universities Osteoarthritis Index pain, stiffness, and function scores of 6.0 +/- 4.0, 3.1 +/- 1.7, and 25 +/- 12 points, respectively. Quality of osteoarthritis care for older adults is suboptimal, particularly with regard to medication safety. Given the high prevalence of osteoarthritis in older age groups, the population impact of any improvement in quality would be substantial. Quality improvement efforts for osteoarthritis should target appropriate use of and counseling regarding medications, as well as underuse of efficacious therapy for osteoarthritis.


An inadequate supply of qualified people will slow the pace of health system change.
Executive recruiters bring a unique perspective on changes in the health system. By knowing the hiring needs of health care organizations, recruiters understand how the organizations are reacting to changes that have already occurred and how their strategies for dealing with the changes set the direction for future trends in the health system. This Issue Brief reports on a roundtable discussion during which a group of executive recruiters considered how health system changes are reflected in recruitment efforts in health care, and what the shifts in recruitment priorities indicate for the future direction and pace of health system change. The roundtable was sponsored by the Center for Studying Health System Change.


Association of Maternal Hypothyroidism With Cardiovascular Diseases in the Offspring.
No previous study has examined the effect of maternal hypothyroidism on a broad spectrum of cardiovascular disease (CVD) endpoints in the offspring. A nationwide population-based cohort study based on the linkage of several Danish nationwide registries was conducted to explore whether maternal hypothyroidism is associated with offspring's CVD. Altogether 1,041,448 singletons born between the 1st of January 1978 and the 31st of December 1998 were investigated from the age of 8 years to the 31st of December 2016. Exposure was maternal diagnosis of hypothyroidism across lifespan and the outcome of interest was a CVD diagnosis in the offspring. Cox regression models were performed to estimate the hazard ratios (HRs) of CVD. Offspring born to mothers with hypothyroidism had an increased risk of CVD (hazard ratios (HR)=1.23, 95% confidence interval (CI): 1.12-1.35), and of several subcategories of CVD including hypertension, arrhythmia, and acute myocardial infarction in offspring. The magnitude of association was the most pronounced in an exposure occur during pregnancy (HR=1.71, 95% CI: 1.10-2.67), which is consistent across all the subgroup analysis, including sibling analysis. Maternal hypothyroidism is associated with an increased risk of CVD in offspring. Thyroid hormone insufficiency during pregnancy may predominantly contribute to the observed associations; however, the effects of a shared genetic background and a time-stable familial environment/lifestyle factors cannot be excluded.


Viral persistence alters CD8 T-cell immunodominance and tissue distribution and results in distinct stages of functional impairment.
Chronic viral infections often result in ineffective CD8 T-cell responses due to functional exhaustion or physical deletion of virus-specific T cells. However, how persisting virus impacts various CD8 T-cell effector functions and influences other aspects of CD8 T-cell dynamics, such as immunodominance and tissue distribution, remains largely unknown. Using different strains of lymphocytic choriomeningitis virus (LCMV), we compared responses to the same CD8 T-cell epitopes during acute or chronic infection. Persistent infection led to a disruption of the normal immunodominance hierarchy of CD8 T-cell responses seen following acute infection and dramatically altered the tissue distribution of LCMV-specific CD8 T cells in lymphoid and nonlymphoid tissues. Most importantly, CD8 T-cell functional impairment occurred in a hierarchical fashion in chronically infected mice. Production of interleukin 2 and the ability to lyse target cells in vitro were the first functions compromised, followed by the ability to make tumor necrosis factor alpha, while gamma interferon production was most resistant to functional exhaustion. Antigen appeared to be the driving force for this loss of function, since a strong correlation existed between the viral load and the level of exhaustion. Further, epitopes presented at higher levels in vivo resulted in physical deletion, while those presented at lower levels induced functional exhaustion. A model is proposed in which antigen levels drive the hierarchical loss of different CD8 T-cell effector functions during chronic infection, leading to distinct stages of functional impairment and eventually to physical deletion of virus-specific T cells. These results have implications for the study of human chronic infections, where similar T-cell deletion and functional dysregulation has been observed.


Structure-based drug design approaches for predicting binding affinities of HIV1 protease inhibitors.
Computational assessment of the binding affinity of enzyme inhibitors prior to synthesis is an important component of computer-assisted drug design (CADD) paradigms. The free energy perturbation (FEP) methodology is the most accurate means of estimating relative binding affinities between two inhibitors. However, due to its complexity and computation-intensive nature, practical applications are restricted to analysis of structurally-related inhibitors. Accordingly, there is a need for methods that enable rapid assessment of large number of structurally-unrelated molecules in a suitably accurate manner. In this review, the FEP method is compared with regression-based methods that employ multivariate models to assess the advantages of each in the estimation of relative binding affinities of inhibitors to an enzyme. Semiquantitative predictions of relative binding free energies of human immunodeficiency virus 1 (HIV1) protease inhibitors are also presented and compared with the corresponding FEP results. The results indicate that the regression-based methods and the FEP method are useful in the semi-quantitative and quantitative assessment of relative binding affinities of enzyme inhibitors, respectively, prior to synthesis.


Neutral Genetic Processes Influence MHC Evolution in Threatened Gopher Tortoises (Gopherus polyphemus).
Levels of adaptive genetic variation influence how species deal with environmental and ecological change, but these levels are frequently inferred using neutral genetic markers. Major histocompatibility complex (MHC) genes play a key role in the adaptive branch of the immune system and have been used extensively to estimate levels of adaptive genetic variation. Parts of the peptide binding region, sites where MHC molecules directly interact with pathogen and self-proteins, were sequenced from a MHC class I (95/441 tortoises) and class II (245/441 tortoises) gene in threatened and nonthreatened populations of gopher tortoises (Gopherus polyphemus), and adaptive genetic variation at MHC genes was compared to neutral genetic variation derived from 10 microsatellite loci (441 tortoises). Genetic diversity at the MHC class II locus and microsatellites was greater in populations in the nonthreatened portion of the gopher tortoise's range (MHC class II difference in mean A = 8.11, AR = 0.79, HO = 0.51, and HE = 0.16; microsatellite difference in mean A = 1.05 and AR = 0.47). Only MHC class II sequences showed evidence of positive selection (dN/dS > 1, Z = 1.81, P = 0.04). Historical gene flow as estimated with Migrate-N was greater than recent migration estimated with BayesAss, suggesting that populations were better connected in the past when habitat was less fragmented. MHC genetic differentiation was correlated with microsatellite differentiation (Mantel r = 0.431, P = 0.001) suggesting neutral genetic processes are influencing MHC evolution, and advantageous MHC alleles could be lost due to genetic drift.


Towards subpercentage uncertainty proton stopping-power mapping via dual-energy CT: Direct experimental validation and uncertainty analysis of a statistical iterative image reconstruction method.
To assess the potential of a joint dual-energy computerized tomography (CT) reconstruction process (statistical image reconstruction method built on a basis vector model (JSIR-BVM)) implemented on a 16-slice commercial CT scanner to measure high spatial resolution stopping-power ratio (SPR) maps with uncertainties of less than 1%. JSIR-BVM was used to reconstruct images of effective electron density and mean excitation energy from dual-energy CT (DECT) sinograms for 10 high-purity samples of known density and atomic composition inserted into head and body phantoms. The measured DECT data consisted of 90 and 140 kVp axial sinograms serially acquired on a Philips Brilliance Big Bore CT scanner without beam-hardening corrections. The corresponding SPRs were subsequently measured directly via ion chamber measurements on a MEVION S250 superconducting synchrocyclotron and evaluated theoretically from the known sample compositions and densities. Deviations of JSIR-BVM SPR values from their theoretically calculated and directly measured ground-truth values were evaluated for our JSIR-BVM method and our implementation of the Hünemohr-Saito (H-S) DECT image-domain decomposition technique for SPR imaging. A thorough uncertainty analysis was then performed for five different scenarios (comparison of JSIR-BVM stopping-power ratio/stopping power (SPR/SP) to International Commission on Radiation Measurements and Units benchmarks; comparison of JSIR-BVM SPR to measured benchmarks; and uncertainties in JSIR-BVM SPR/SP maps for patients of unknown composition) per the Joint Committee for Guides in Metrology and the Guide to Expression of Uncertainty in Measurement, including the impact of uncertainties in measured photon spectra, sample composition and density, photon cross section and I-value models, and random measurement uncertainty. Estimated SPR uncertainty for three main tissue groups in patients of unknown composition and the weighted proportion of each tissue type for three proton treatment sites were then used to derive a composite range uncertainty for our method. Mean JSIR-BVM SPR estimates deviated by less than 1% from their theoretical and directly measured ground-truth values for most inserts and phantom geometries except for high-density Delrin and Teflon samples with SPR error relative to proton measurements of 1.1% and -1.0% (head phantom) and 1.1% and -1.1% (body phantom). The overall root-mean-square (RMS) deviations over all samples were 0.39% and 0.52% (head phantom) and 0.43% and 0.57% (body phantom) relative to theoretical and directly measured ground-truth SPRs, respectively. The corresponding RMS (maximum) errors for the image-domain decomposition method were 2.68% and 2.73% (4.68% and 4.99%) for the head phantom and 0.71% and 0.87% (1.37% and 1.66%) for the body phantom. Compared to H-S SPR maps, JSIR-BVM yielded 30% sharper and twofold sharper images for soft tissues and bone-like surrogates, respectively, while reducing noise by factors of 6 and 3, respectively. The uncertainty (coverage factor k = 1) of the DECT-to-benchmark values comparison ranged from 0.5% to 1.5% and is dominated by scanning-beam photon-spectra uncertainties. An analysis of the SPR uncertainty for patients of unknown composition showed a JSIR-BVM uncertainty of 0.65%, 1.21%, and 0.77% for soft-, lung-, and bony-tissue groups which led to a composite range uncertainty of 0.6-0.9%. Observed JSIR-BVM SPR estimation errors were all less than 50% of the estimated k = 1 total uncertainty of our benchmarking experiment, demonstrating that JSIR-BVM high spatial resolution, low-noise SPR mapping is feasible and is robust to variations in the geometry of the scanned object. In contrast, the much larger H-S SPR estimation errors are dominated by imaging noise and residual beam-hardening artifacts. While the uncertainties characteristic of our current JSIR-BVM implementation can be as large as 1.5%, achieving < 1% total uncertainty is feasible by improving the accuracy of scanner-specific scatter-profile and photon-spectrum estimates. With its robustness to beam-hardening artifact, image noise, and variations in phantom size and geometry, JSIR-BVM has the potential to achieve high spatial-resolution SPR mapping with subpercentage accuracy and estimated uncertainty in the clinical setting.


Temporal variations of polybrominated dibenzo-p-dioxin and methoxylated diphenyl ether concentrations in fish revealing large differences in exposure and metabolic stability.
The concentrations of polybrominated dibenzo-p-dioxins (PBDDs) and polybrominated methoxylated diphenyl ethers (MeO-PBDEs) were investigated in perch (Perca fluviatilis) collected from a Baltic Sea background contaminated area between 1990 and 2005. No temporal trend was found, but large variations were observed - up to 5-fold and 160-fold differences in MeO-PBDE and PBDD concentrations, respectively - between consecutive years, suggesting that retention of these compounds, particularly the PBDDs, is limited. Examination of the congener profiles using principal component analysis (PCA) and correlation analysis indicated that MeO-PBDEs without adjacent substituents (6-MeO-BDE47) or with two adjacent substituents (2'-MeO-BDE68 and 6-MeO-BDE90) are retained more than MeO-PBDEs with three adjacent substituents (6-MeO-BDE85 and 6-MeO-BDE99) and that 1,3,6,8-tetraBDD and 1,3,7,9-tetraBDD are retained more than the other PBDDs which have vicinal hydrogen. Debromination could explain the limited retention of 6-MeO-PBDE85 and 6-MeO-BDE99 and the absence of 2-MeO-BDE123 and 6-MeO-BDE137, and cytochrome P-450 mediated oxidation could explain the limited retention of PBDDs containing vicinal hydrogen. The levels of organobromines, especially MeO-PBDEs, were found to covary with water conditions related to primary production, for example temperature, depth visibility, and inorganic nutrient concentrations, which also favor fish productivity. The results suggest natural production of MeO-PBDEs and PBDDs and imply that they fluctuate considerably over time, as do common marine toxins in fish. Thus, assessments of human and environmental risk should consider both the average and peak concentrations of these contaminants in marine biota.


[Sensitivity of various metabolic processes to adrenaline and insulin experimental thyrotoxicosis].
The fasting hypoglycemia and hyperlipocidemia were observed in rabbits fed with massive doses of thyroidine. The hypoglycemic effect of insulin was diminished but its hypolipocidemic action rose. In the isolated diaphragm of rats given massive doses of 1-thyroxine insulin produced a more pronounced effect on glucose uptake than in the muscle of control animals. The hyperglycemic action of epinephrine in thyrotoxic rabbits remained unaltered, but its hyperlipocidemic effect proved to be less than in control animals. The in vitro effect of epinephrine on the adipose tissue lipolytic activity of severe thyrotoxic rats was less than that in control rats.


Abnormal fatty alcohol metabolism in cultured keratinocytes from patients with Sjögren-Larsson syndrome.
Sjögren-Larsson syndrome (SLS) is an inherited neurocutaneous disorder characterized by ichthyosis, mental retardation, spasticity, and deficient activity of fatty aldehyde dehydrogenase (FALDH). FALDH is an enzyme component of fatty alcohol:NAD oxidoreductase (FAO), which is necessary for fatty alcohol metabolism. To better understand the biochemical basis for the cutaneous symptoms in this disease, we investigated lipid metabolism in cultured keratinocytes from SLS patients. Enzyme activities of FALDH and FAO in SLS cells were <10% of normal. SLS keratinocytes accumulated 45-fold more fatty alcohol (hexadecanol, octadecanol, and octadecenol) than normal, whereas wax esters and 1-O-alkyl-2,3-diacylglycerols were increased by 5.6-fold and 7.5-fold, respectively. SLS keratinocytes showed a reduced incorporation of radioactive octadecanol into fatty acid (24% of normal) and triglyceride (13% of normal), but incorporation into wax esters and 1-O-alkyl-2,3-diacylglycerol was increased by 2.5-fold and 2.8-fold, respectively. Our results indicate that FALDH deficiency in SLS keratinocytes causes the accumulation and diversion of fatty alcohol into alternative biosynthetic pathways. The striking lipid abnormalities in cultured SLS keratinocytes are distinct from those seen in fibroblasts and may be related to the stratum corneum dysfunction and ichthyosis in SLS.


[Prospective memory and its theoretical considerations].
Prospective memory involves memory for future intentions in our everyday lives and it is one of the hot topics in current memory research. We reexamined what is prospective memory and discussed how intentions had been conceived of in the history of psychology. We emphasize the necessity of classifying a form of remembering intentions into the self-initiated existence-remembrance and its content-remembrance. Many studies on prospective memory in recent decades were reviewed with respect to the following perspectives: (1) how intentions are recollected, (2) what is remembering intentions based on, (3) how we attempt to avoid forgetting to do things, and (4) what is neuropsychological mechanism of remembering intentions. Requirements for experimental research on prospective memory are summarized and future directions are discussed.


NO decreases phosphorylation of focal adhesion proteins via reduction of Ca in rat aortic smooth muscle cells.
Our laboratory has previously reported that the antimitogenic effect of nitric oxide (NO) in primary cultures of rat aortic smooth muscle cells may be attributed to activation of protein tyrosine phosphatase and dephosphorylation of protein phosphotyrosine [G.S. Dhaunsi, C. Matthews, K. Kaur, and A. Hassid, Am, J. Physiol. 272 (Heart Circ. Physiol. 41): H1342-H1349, 1997]. The goal of the current study was to investigate the role of cytoplasmic Ca in this process and to identify protein substrates that are dephosphorylated by treatment with NO. Treatment of primary rat aortic smooth muscle cell cultures with the NO donor S-nitroso-N-acetylpenicillamine (SNAP) decreased cytoplasmic Ca levels and elicited phosphotyrosine dephosphorylation. Both effects were mimicked by the extracellular and intracellular Ca chelators ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), respectively, and by the Ca channel blocker nifedipine. Conversely, elevation of cytoplasmic Ca via the use of the Ca ionophore A-23187 or high extracellular K+ prevented or attenuated SNAP-induced dephosphorylation. Both BAPTA and nifedipine also decreased DNA synthesis, providing further evidence to link dephosphorylation to antimitogenesis. Two of the proteins dephosphorylated by treatment of cells with NO or EGTA were identified as the focal adhesion proteins, cortactin and paxillin. These results indicate that NO-induced dephosphorylation of protein phosphotyrosine is mediated by reduction of cytoplasmic Ca and suggest that dephosphorylation of focal adhesion proteins may be of relevance to the antimitogenic effect of NO.


Regulation of calpain and calpastatin in differentiating myoblasts: mRNA levels, protein synthesis and stability.
Calpain (Ca(2+)-dependent intracellular protease)-induced proteolysis has been considered to play a role in myoblast fusion to myotubes. We found previously that calpastatin (the endogenous inhibitor of calpain) diminishes transiently during myoblast differentiation. To gain information about the regulation of calpain and calpastatin in differentiating myoblasts, we evaluated the stability and synthesis of calpain and calpastatin, and measured their mRNA levels in L8 myoblasts. We show here that mu-calpain and m-calpain are stable, long-lived proteins in both dividing and differentiating L8 myoblasts. Calpain is synthesized in differentiating myoblasts, and calpain mRNA levels do not change during differentiation. In contrast, calpastatin (though also a long-lived protein in myoblasts), is less stable in differentiating myoblasts than in the dividing cells, and its synthesis is inhibited upon initiation of differentiation. Inhibition of calpastatin synthesis is followed by a diminution in calpastatin mRNA levels. A similar calpastatin mRNA diminution is observed upon drug-induced inhibition of protein translation. On the other hand, transforming growth factor beta (which inhibits differentiation) allows calpastatin synthesis and prevents the diminution in calpastatin mRNA. The overall results suggest that at the onset of myoblast differentiation, calpastatin is regulated mainly at the level of translation and that an inhibition of calpastatin synthesis leads to the decrease in its mRNA stability. The existing calpastatin then diminishes, resulting in decreased calpastatin activity in the fusing myoblasts, allowing calpain activation and protein degradation required for fusion.


Dynamic Electromechanical Hydrogel Matrices for Stem Cell Culture.
Hydrogels have numerous biomedical applications including synthetic matrices for cell culture and tissue engineering. Here we report the development of hydrogel based multifunctional matrices that not only provide three-dimensional structural support to the embedded cells but also can simultaneously provide potentially beneficial dynamic mechanical and electrical cues to the cells. A unique aspect of these matrices is that they undergo reversible, anisotropic bending dynamics in an electric field. The direction and magnitude of this bending can be tuned through the hydrogel crosslink density while maintaining the same electric potential gradient, allowing control over the mechanical strain imparted to the cells in a three-dimensional environment. The conceptual design of these hydrogels was motivated through theoretical modeling of the osmotic pressure changes occurring at the gel-solution interfaces in an electric field. These electro-mechanical matrices support survival, proliferation, and differentiation of stem cells. Thus, these new three-dimensional in vitro synthetic matrices, which mimic multiple aspects of the native cellular environment, take us one step closer to in vivo systems.


Sexual selection on brain size in shorebirds (Charadriiformes).
Natural selection is considered a major force shaping brain size evolution in vertebrates, whereas the influence of sexual selection remains controversial. On one hand, sexual selection could promote brain enlargement by enhancing cognitive skills needed to compete for mates. On the other hand, sexual selection could favour brain size reduction due to trade-offs between investing in brain tissue and in sexually selected traits. These opposed predictions are mirrored in contradictory relationships between sexual selection proxies and brain size relative to body size. Here, we report a phylogenetic comparative analysis that highlights potential flaws in interpreting relative brain size-mating system associations as effects of sexual selection on brain size in shorebirds (Charadriiformes), a taxonomic group with an outstanding diversity in breeding systems. Considering many ecological effects, relative brain size was not significantly correlated with testis size. In polyandrous species, however, relative brain sizes of males and females were smaller than in monogamous species, and females had smaller brain size than males. Although these findings are consistent with sexual selection reducing brain size, they could also be due to females deserting parental care, which is a common feature of polyandrous species. Furthermore, our analyses suggested that body size evolved faster than brain size, and thus the evolution of body size may be confounding the effect of the mating system on relative brain size. The brain size-mating system association in shorebirds is thus not only due to sexual selection on brain size but rather, to body size evolution and other multiple simultaneous effects.


Cyclooxygenase-2 inhibition improves amyloid-beta-mediated suppression of memory and synaptic plasticity.
Non-steroidal anti-inflammatory agents (NSAIDs) are associated with a marked reduction in the risk of developing Alzheimer's disease, a form of dementia characterized by the accumulation of amyloid plaques containing the amyloid-beta protein (Abeta). Studies of the effects of NSAIDs upon the inflammatory response surrounding amyloid plaques and upon the generation of Abeta from the amyloid precursor protein (APP) have led to two proposed mechanisms by which NSAIDs may protect against Alzheimer's disease: one, the selective lowering of Abeta42 by a subset of NSAIDs; and two, the reduction of inflammation. Although Alzheimer's disease is a disorder of brain and synaptic function, the effects of NSAIDs on Abeta-mediated suppression of synaptic plasticity and memory function have never been reported. We therefore investigated how three different NSAIDs, chosen for their distinct effects on Abeta42 production and the inhibition of the cyclooxygenase (COX) isoenzymes, COX-1 and COX-2, affect memory function and synaptic plasticity. By focusing upon brain and synapse function, we made novel observations about the effects of NSAIDs on Abeta-mediated neural processes. Here we report that the selective inhibition of COX-2, but not COX-1, acutely prevented the suppression of hippocampal long-term plasticity (LTP) by Abeta. The non-selective NSAIDs, ibuprofen and naproxen, and a selective COX-2 inhibitor, MF-tricyclic, each restored memory function in Tg2576 mice over-expressing APP, and also blocked Abeta-mediated inhibition of LTP. There was no advantage of ibuprofen, a selective Abeta42-lowering agent (SALA), over the non-SALAs, naproxen and MF-tricyclic. The beneficial effects on memory did not depend upon lowered levels of Abeta42 or the inflammatory cytokines, tumour necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta). Intriguingly, improved memory function was inversely related to prostaglandin E2 (PGE2) levels. Conversely, exogenous PGE2 prevented the restorative effects of COX-2 inhibitors on LTP. The data indicate that the inhibition of COX-2 blocks Abeta-mediated suppression of LTP and memory function, and that this block occurs independently of reductions in Abeta42 or decreases in inflammation. The results lead us to propose a third possible mechanism by which NSAIDs may protect against Alzheimer's disease, involving the blockade of a COX-2-mediated PGE2 response at synapses.


Subsidence of Additively-Manufactured Cages in Foam Substrates: Effect of Contact Topology.
Subsidence of implants into bone is a major source of morbidity. The underlying mechanics of the phenomenon are not clear, but are likely related to interactions between contact stresses and the underlying porous trabecular bone structure. To gain insight into these interactions, we studied the penetration of three-dimensional (3D)-printed indenters with systematically varying geometries into Sawbones® foam substrates and isolated the effects of contact geometry from those of overall contact size and area. When size, contact area, and indented material stiffness and strength are controlled for, we show that resistance to penetration is in fact a function of topology only. Indenters with greater line contact lengths support higher subsidence loads in compression. These results have direct implications for the design of implants to resist subsidence into bone.


Aspirin-triggered resolvin D1 reduces mucosal inflammation and promotes resolution in a murine model of acute lung injury.
Acute lung injury (ALI) is a severe illness with excess mortality and no specific therapy. Protective actions were recently uncovered for docosahexaenoic acid-derived mediators, including D-series resolvins. Here, we used a murine self-limited model of hydrochloric acid-induced ALI to determine the effects of aspirin-triggered resolvin D1 (AT-RvD1; 7S,8R,17R-trihydroxy-4Z,9E,11E,13Z,15E,19Z-docosahexaenoic acid) on mucosal injury. RvD1 and its receptor ALX/FPR2 were identified in murine lung after ALI. AT-RvD1 (~0.5-5 μg kg(-1)) decreased peak inflammation, including bronchoalveolar lavage fluid (BALF) neutrophils by ~75%. Animals treated with AT-RvD1 had improved epithelial and endothelial barrier integrity and decreased airway resistance concomitant with increased BALF epinephrine levels. AT-RvD1 inhibited neutrophil-platelet heterotypic interactions by downregulating both P-selectin and its ligand CD24. AT-RvD1 also significantly decreased levels of BALF pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, Kupffer cells, and tumor necrosis factor-α, and decreased nuclear factor-κB-phosphorylated p65 nuclear translocation. Taken together, these findings indicate that AT-RvD1 displays potent mucosal protection and promotes catabasis after ALI.


Nutritional, dietary and postprandial oxidative stress.
Nutritional, or dietary oxidative stress denotes a disturbance of the redox state resulting from excess oxidative load or from inadequate nutrient supply favoring prooxidant reactions. Low intake or impaired availability of dietary antioxidants including vitamins E and C, carotenoids, polyphenols, and other micronutrients (e.g., selenium) weakens the antioxidant network. Postprandial oxidative stress, as a subform of nutritional oxidative stress, ensues from sustained postprandial hyperlipidemia and/or hyperglycemia and is associated with a higher risk for atherosclerosis, diabetes, and obesity. In Western societies, a significant part of the day is spent in the postprandial state. Unsaturated fatty acids incorporated into LDL and oxidized LDL are an atherogenic factor. Lipid hydroperoxides present in the diet are absorbed, contributing to the prooxidant load. In hyperlipidemic and hyperglycemic subjects, endothelium-dependent vasodilation is impaired in the postprandial state, making postprandial oxidative stress an important factor modulating cardiovascular risk. Postprandial oxidative stress is attenuated when dietary antioxidants are supplied together with a meal rich in oxidized or oxidizable lipids. Ingestion of dietary polyphenols, e.g., from wine, cocoa, or tea, improves endothelial dysfunction and lowers the susceptibility of LDL lipids to oxidation. Polyphenols affect endothelial function not solely as antioxidants but also as modulatory signaling molecules.


A 2D gel electrophoresis-based snapshot of the phosphoproteome in the cyanobacterium Synechocystis sp. strain PCC 6803.
Cyanobacteria are photoautotrophic prokaryotes that occur in highly variable environments. Protein phosphorylation is one of the most widespread means to adjust cell metabolism and gene expression to the demands of changing growth conditions. Using a 2D gel electrophoresis-based approach and a phosphoprotein-specific dye, we investigated the protein phosphorylation pattern in cells of the model cyanobacterium Synechocystis sp. strain PCC 6803. The comparison of gels stained for total and phosphorylated proteins revealed that approximately 5 % of the protein spots seemed to be phosphoproteins, from which 32 were identified using MALDI-TOF MS. For eight of them the phosphorylated amino acid residues were mapped by subsequent mass spectrometric investigations of isolated phosphopeptides. Among the phosphoproteins, we found regulatory proteins, mostly putative anti-sigma factor antagonists, and proteins involved in translation. Moreover, a number of enzymes catalysing steps in glycolysis or the Calvin-Benson cycle were found to be phosphorylated, implying that protein phosphorylation might represent an important mechanism for the regulation of the primary carbon metabolism in cyanobacterial cells.


Impedance analyser module for EIT and spectroscopy using undersampling.
In this paper we present the concept, the design and the test procedure for a DSP-based high-precision and high-performance wide-band (up to 10 MHz) bioimpedance analyser module for application in EIT or bioimpedance spectroscopy. The module implements a digital concept with appropriate signal conditioning hardware for voltage and current measurement, early signal digitization and subsequent digital signal processing in order to calculate the components of impedance (or admittance). At low frequencies, the module utilizes the conventional direct conversion method, whereas at high frequencies the undersampling technique is used. The advantages of the described system are the following: (a) the frequency range is extended to higher frequencies, (b) the number of data sampled per time interval is significantly reduced, and (c) the current consumption and the costs of the ADCs can be significantly reduced. The validation procedure is performed by comparing the measured and theoretical values of the magnitude and the phase of the impedance for a commonly used tissue model. The module offers an accuracy of better than 0.012% for the magnitude of impedance and better than 0.02 degrees for the phase.


Evolution of discrete phenotypes from continuous norms of reaction.
Discrete phenotypic variation often involves threshold expression of a trait with polygenic inheritance. How such discrete polyphenisms evolve starting from continuously varying phenotypes has received little theoretical attention. We model the evolution of sigmoid norms of reaction in response to variation in an underlying trait or in a continuous environment to identify conditions for the evolution of discontinuity. For traits with expression depending on a randomly varying underlying factor, such as developmental noise, polyphenism is unstable under constant phenotypic selection for two selective peaks, and reaction norm evolution results in a phenotypic distribution concentrated at only one peak. But with frequency-dependent selection between two adaptive peaks, a steep threshold maintaining polyphenism can evolve. For inducible plastic traits with expression conditioned on an environmental variable that also affects phenotypic selection, the steepness of the evolved reaction norm depends both on the differentiation of the environment in time or space and on its predictability between development and selection. Together with recent measurements of genetic variance of threshold steepness, these predictions suggest that quasi-discrete phenotypic variation may often evolve from continuous norms of reactions rather than being an intrinsic property of development.


Regulation of ATP supply during muscle contraction: theoretical studies.
The dynamic computer model of oxidative phosphorylation developed previously and successfully tested for large-scale changes in fluxes and metabolite concentrations was used to study the question of how the rate of ATP production by oxidative phosphorylation is adjusted to meet the energy demand during muscle contraction, which causes a great increase in ATP consumption in relation to the resting state. The changes in the respiration rate and ATP/ADP ratio after the onset of maximal work measured experimentally were compared with simulated changes in the respiration rate and ATP/ADP in several different cases, assuming direct activation of different steps by an external effector. On the basis of the computer simulations performed, it was possible to conclude which enzymes/metabolic blocks should be directly activated to cause the experimentally observable changes in fluxes and metabolite concentrations. The theoretical results obtained suggest that the parallel direct activation of actinomyosin-ATP-ase and oxidative phosphorylation by an external effector (for example calcium ions) is the main mechanism responsible for fitting of ATP production to ATP consumption, while the negative feedback via an increase in ADP concentration (decrease in ATP/ADP), which indirectly activates the ATP supply, plays only a minor role. Additionally, the conclusion is drawn that most of the oxidative phosphorylation steps should be directly activated in order to explain the observed changes in the respiration rate and ATP/ADP ratio (and also in other parameters) during muscle contraction. It is suggested that there should exist a universal external activator/regulatory mechanism which causes a parallel stimulation of different enzymes/processes. A possible nature of such an activator is shortly discussed.


The influence of ascorbic acid on the growth of solid tumors in mice and on tumor control by X-irradiation.
Swiss mice drank either distilled water or 0.1% ascorbic acid in distilled water for one week prior to and during the experiments. Solid tumors were induced by injecting Ehrlich ascites tumor cells i.m. into the hind limb of Swiss male and female mice. It was found that the tumor growth was significantly faster in the mice that were drinking distilled water. Beginning two days after tumor cell injection, the tumor bearing limbs were irradiated every 24 h as follows: a) 6 exposures of 400 R, 500 R, 600 R, or 700 R each, b) 10 exposures of 400 R each, and c) 11 exposures of 400 R each. Our results indicated that when 6 exposures of 700 R each were given every 24 h to mice drinking distilled water, about 80-85% tumor control was achieved; the percent tumor cure was even better in mice drinking ascorbic acid in water than in mice drinking water.


Design of blocking filters of any narrow bandwidth.
A design approach is described to achieve spectral blocking filters of any spectral width and optical density for narrow blocking bands. The ratio of the thickness of the high-index material to the low-index material in the layer pairs is adjusted to obtain the desired bandwidth in the first-harmonic reflection band. The number of layer pairs is adjusted to provide the required optical density. Equations are provided for estimating the ratios and number of layer pairs needed to achieve a given bandwidth and optical density. This approach can be useful for laser line blocking filters, night vision filters, and other general applications.


Origins of the selectivity for borylation of primary over secondary C-H bonds catalyzed by Cp*-rhodium complexes.
Detailed experimental and computational studies of the high selectivity for functionalization of primary over secondary sp(3) C-H bonds in alkanes by borane reagents catalyzed by Cp*Rh complexes are reported. Prior studies have shown that Cp*Rh(X)(Bpin) (X = H or Bpin), generated from Cp*Rh(H)(2)(Bpin)(2) and Cp*Rh(H)(2)(Bpin)(3), are likely intermediates in these catalytic reactions. To allow analysis of the system by H/D exchange, the current studies focused on reactions of Cp*Rh(D)(2)(Bpin)(2) through the 16-electron species Cp*Rh(D)(Bpin). Density functional theory (DFT) calculations of the reaction between Cp*Rh(H)(BO(2)C(2)H(4)) and the primary and secondary C-H bonds of propane indicate that the lowest energy pathway for C-H bond cleavage occurs to form an isomer in which the alkyl and boryl groups are trans to each other, while the lowest energy pathway for functionalization of the primary C-H bond occurs by formation of the isomer in which these two groups are cis to each other. The barrier for formation of the rhodium complex by cleavage of secondary C-H bonds is higher than that by cleavage of primary C-H bond. The alkyl intermediates are formed reversibly, and steric effects cause the barrier for B-C bond formation from the secondary alkyl intermediate to be higher than that from the primary alkyl intermediate. Experimental studies are consistent with this computational analysis. H/D exchange occurs between (Cp*d(15))Rh(D)(2)(Bpin)(2) and n-octane, indicating that C-H bond cleavage occurs reversibly and occurs faster at primary over secondary C-H bonds. The observation of small amounts of H/D exchange into the secondary C-H bonds of linear alkanes and the clear observation of H/D exchange into the secondary positions of cyclic alkanes without formation of products from functionalization are consistent with the high barrier calculated for B-C bond formation from the secondary alkyl intermediate. A series of kinetic experiments are consistent with a mechanism for H/D exchange between (Cp*d(15))Rh(D)(2)(Bpin)(2) and n-octane occurring by dissociation of borane-d(1) to form (Cp*d(15))Rh(D)(Bpin). Thus, the origin of the selectivity for borylation of primary over secondary C-H bonds is due to the cumulative effects of selective C-H bond cleavage and selective C-B bond formation.


Pigment epithelium-derived factor binds to hyaluronan. Mapping of a hyaluronan binding site.
Pigment epithelium-derived factor (PEDF) is a multifunctional serpin with antitumorigenic, antimetastatic, and differentiating activities. PEDF is found within tissues rich in the glycosaminoglycan hyaluronan (HA), and its amino acid sequence contains putative HA-binding motifs. We show that PEDF coprecipitation with glycosaminoglycans in media conditioned by human retinoblastoma Y-79 cells decreased after pretreatments with hyaluronidase, implying an association between HA and PEDF. Direct binding of human recombinant PEDF to highly purified HA was demonstrated by coprecipitation in the presence of cetylpyridinium chloride. Binding of PEDF to HA was concentration-dependent and saturable. The PEDF-HA interactions were sensitive to increasing NaCl concentrations, indicating an ionic nature of these interactions and having affinity higher than PEDF-heparin. Competition assays showed that PEDF can bind heparin and HA simultaneously. PEDF chemically modified with fluorescein retained the capacity for interacting with HA but lacked heparin affinity, suggesting one or more distinct HA-binding regions on PEDF. The HA-binding region was examined by site-directed mutagenesis. Single-point and cumulative alterations at basic residues within the putative HA-binding motif K189A/K191A/R194A/K197A drastically reduced the HA-binding activity without affecting heparin- or collagen I binding of PEDF. Cumulative alterations at sites critical for heparin binding (K146A/K147A/R149A) decreased HA affinity but not collagen I binding. Thus these clusters of basic residues (BXBXXBXXB and BX3AB2XB motifs) in PEDF are functional regions for binding HA. In the spatial PEDF structure they are located in distinct areas away from the collagen-binding site. The HA-binding activity of PEDF may contribute to deposition in the extracellular matrix and to its reported antitumor/antimetastatic effects.


Central role for MCP-1/CCL2 in injury-induced inflammation revealed by in vitro, in silico, and clinical studies.
The translation of in vitro findings to clinical outcomes is often elusive. Trauma/hemorrhagic shock (T/HS) results in hepatic hypoxia that drives inflammation. We hypothesize that in silico methods would help bridge in vitro hepatocyte data and clinical T/HS, in which the liver is a primary site of inflammation. Primary mouse hepatocytes were cultured under hypoxia (1% O2) or normoxia (21% O2) for 1-72 h, and both the cell supernatants and protein lysates were assayed for 18 inflammatory mediators by Luminex™ technology. Statistical analysis and data-driven modeling were employed to characterize the main components of the cellular response. Statistical analyses, hierarchical and k-means clustering, Principal Component Analysis, and Dynamic Network Analysis suggested MCP-1/CCL2 and IL-1α as central coordinators of hepatocyte-mediated inflammation in C57BL/6 mouse hepatocytes. Hepatocytes from MCP-1-null mice had altered dynamic inflammatory networks. Circulating MCP-1 levels segregated human T/HS survivors from non-survivors. Furthermore, T/HS survivors with elevated early levels of plasma MCP-1 post-injury had longer total lengths of stay, longer intensive care unit lengths of stay, and prolonged requirement for mechanical ventilation vs. those with low plasma MCP-1. This study identifies MCP-1 as a main driver of the response of hepatocytes in vitro and as a biomarker for clinical outcomes in T/HS, and suggests an experimental and computational framework for discovery of novel clinical biomarkers in inflammatory diseases.


The timing and quality of antenatal care received by women attending a primary care centre in Iquitos, Peru: A facility exit survey.
Maternal mortality is high in Loreto, Peru, but can be reduced by high quality antenatal care. Indicators for the quality of antenatal care received include the timing (with respect to gestational age) and number of antenatal appointments attended, the delivery of antenatal services and health information, and women's perceptions about their care. This study investigated these indicators amongst women receiving antenatal care in predominantly the San Juan Bautista district of Iquitos, Loreto. This was to identify areas for improvement through comparison with antenatal guidelines published by the Ministry of Health, Peru, and the World Health Organization. A total of 134 women were recruited at the Centro de Salud, San Juan-a primary care centre in Iquitos. Information about the delivery of antenatal services and the number of and gestational ages at appointments attended was collected from 121/134 women's hand-held antenatal cards. The delivery of health information and women's perceptions about their antenatal care were investigated through questionnaires (133/134 completed). Descriptive statistics, such as frequencies and valid percentages, were determined. Hand-held antenatal cards revealed that 52.9% of participants began their antenatal care in the first trimester. Compared to national guidelines, 42.1% attended appointments at recommended gestational ages and no women received all recommended antenatal services. Most women received information about identifying complications in pregnancy and health and lifestyle topics. Over 85% of women reported satisfaction with their antenatal care. Timely antenatal attendance and delivery of services should be encouraged to meet national and global standards. Although all services were not delivered in a combined manner according to national guidelines, individual services were mostly delivered to a high standard and therefore a high proportion of women were satisfied with their antenatal care.


Killing mechanism of Listeria monocytogenes in activated macrophages as determined by an improved assay system.
Exposure of Listeria monocytogenes to gentamicin 5 mg/L for 4 h resulted in the killing of most extracellular bacteria, but had no effect on the survival of bacteria inside macrophages. Higher concentrations of gentamicin caused a reduction in the number of intracellular bacteria. This effect was associated with cellular uptake of gentamicin, but was unaffected by activation of macrophages by interferon-gamma and lipopolysaccharide. In experiments in which exposure to gentamicin 5 mg/L for 4 h was used to kill extracellular bacteria, killing by activated macrophages was impaired when O2- production was inhibited by superoxide dismutase, but not when nitric oxide production was blocked by NG-monomethyl-L-arginine. These data suggest that the reactive oxygen intermediates are more important than nitric oxide in the killing of L. monocytogenes, at least in macrophages activated in vitro.


Prostaglandin independence of kinin-stimulated renin release.
Bradykinin can increase prostaglandin synthesis and also stimulate renin release in vitro. Because prostaglandins also stimulate renin, studies were performed to determine whether bradykinin stimulation of renin is a function of prostaglandin synthesis. Isolated glomeruli with attendant arteriolar attachments were harvested from rat kidneys and superfused. The effluent was analyzed for renin, prostaglandins E2 and I2 (6-keto-PGF1 alpha). Bradykinin (10(-5) M) increased renin by 50% with a concomitant increase in prostacyclin (PGI2) but not in prostaglandin E2 (PGE2). The cyclooxygenase inhibitor meclofenamate (1.6 X 10(-5) M) inhibited bradykinin-induced PGI2 synthesis but not the concurrent increase in renin release. Additionally, neither the phospholipase inhibitor quinacrine (10(-2) M) nor the prostacyclin synthetase inhibitor 9,11-azoprosta-5,13-dienoic acid (Azo analogue-1) (5.67 X 10(-6) M) eliminated bradykinin-induced renin release. Superfusion with calcium-free media and EDTA increased basal renin release 2.5-fold, and bradykinin stimulated a twofold increase in renin release. Neither a high (10(-2) M) media calcium nor the calcium channel blocker nifedipine (10(-6) M) eliminated bradykinin stimulation of renin. These results suggest that bradykinin stimulation of renin is at least partially independent of prostaglandin synthesis and that bradykinin must act by some prostaglandin-independent pathway to induce renin release from isolated glomeruli.


Evidence based management guideline for the COVID-19 pandemic - Review article.
COVID-19 has now been declared a pandemic. To date, COVID-19 has affected over 2.5 million people worldwide, resulting in over 170,000 reported deaths. Numerous preventative strategies and non-pharmaceutical interventions have been employed to mitigate the spread of disease including careful infection control, the isolation of patients, and social distancing. Management is predominantly focused on the provision of supportive care, with oxygen therapy representing the major treatment intervention. Medical therapy involving corticosteroids and antivirals have also been encouraged as part of critical management schemes. However, there is at present no specific antiviral recommended for the treatment of COVID-19, and no vaccine is currently available. Despite the strategic implementation of these measures, the number of new reported cases continues to rise at a profoundly alarming rate. As new findings emerge, there is an urgent need for up-to-date management guidelines. In response to this call, we review what is currently known regarding the management of COVID-19, and offer an evidence-based review of current practice.


Study of the muscarinic receptor subtypes in N1E 115 mouse neuroblastoma cells.
Muscarinic receptors in N1E 115 mouse neuroblastoma cells were characterized by competition binding experiments using three agonists and five antagonists, including 4-DAMP and AF-DX 116, and by studying the effect of agonist stimulation on the cellular cAMP and cGMP content. The results of the binding studies with the antagonists suggest that only one single homogeneous binding site of the M1 muscarinic receptor subtype is present. For the binding with the agonists, two binding sites were detected, one with high affinity for the ligand (between 53 and 77% of the total binding sites depending on the agonist) and one with low affinity. In contrast to the results obtained with the binding experiments using antagonists, the study of the cellular cyclic nucleotide response upon carbachol stimulation suggested the presence of both the M1 and M2 subtypes as there was an increase in cyclic GMP concentration while at the same time, the prostaglandin-stimulated synthesis of cyclic AMP was inhibited. Considering both binding and functional data we suggest that in N1E 115 cells a majority of M1 and a minority of M2 muscarinic receptors are present; there is no evidence for the presence of M3 muscarinic receptors.


Risk factors for rehospitalization for acute coronary syndromes and unplanned revascularization following acute myocardial infarction.
Rehospitalizations for acute coronary syndromes (ACS) and coronary revascularization after an acute myocardial infarction (AMI) are not only common and costly but can also impact patients' quality of life. In contrast to mortality and all-cause readmissions, little insight is available into risk factors associated with ACS and revascularization after AMI. In a multicenter AMI registry, we examined the rates and predictors of rehospitalizations for ACS and revascularization within the year after AMI among 3283 patients. Staged revascularization procedures were excluded. Kaplan-Meier estimated rates of rehospitalization due to ACS and revascularization were 6.8% and 4.1%, respectively. In hierarchical, multivariable models, the strongest predictors of rehospitalization for ACS were coronary artery bypass graft prior to AMI hospitalization (hazard ratio [HR] 2.12, 95% CI 1.45 to 3.10), female sex (HR 1.67, 95% CI 1.23 to 2.25), and in-hospital PCI (HR 1.85, 95% CI 1.28 to 2.69). The strongest predictors of subsequent revascularization were multivessel disease (HR 2.89, 95% CI 1.90 to 4.39) and in-hospital percutaneous coronary intervention with a bare metal stent (HR 2.08, 95% CI 1.19 to 3.63). The Global Registry of Acute Coronary Events mortality risk score was not associated with the risk of rehospitalization for ACS or revascularization. Unique characteristics are associated with admissions for ACS and revascularization, as compared with survival. These multivariable risk predictors may help identify patients at high risk for ACS and revascularization, in whom intensification of secondary prevention therapies or closer post-AMI follow-up may be warranted.


Kidney-based in vitro models for drug-induced toxicity testing.
The kidney is frequently involved in adverse effects caused by exposure to foreign compounds, including drugs. An early prediction of those effects is crucial for allowing novel, safe drugs entering the market. Yet, in current pharmacotherapy, drug-induced nephrotoxicity accounts for up to 25% of the reported serious adverse effects, of which one-third is attributed to antimicrobials use. Adverse drug effects can be due to direct toxicity, for instance as a result of kidney-specific determinants, or indirectly by, e.g., vascular effects or crystals deposition. Currently used in vitro assays do not adequately predict in vivo observed effects, predominantly due to an inadequate preservation of the organs' microenvironment in the models applied. The kidney is highly complex, composed of a filter unit and a tubular segment, together containing over 20 different cell types. The tubular epithelium is highly polarized, and the maintenance of this polarity is critical for optimal functioning and response to environmental signals. Cell polarity is dependent on communication between cells, which includes paracrine and autocrine signals, as well as biomechanic and chemotactic processes. These processes all influence kidney cell proliferation, migration, and differentiation. For drug disposition studies, this microenvironment is essential for prediction of toxic responses. This review provides an overview of drug-induced injuries to the kidney, details on relevant and translational biomarkers, and advances in 3D cultures of human renal cells, including organoids and kidney-on-a-chip platforms.


Oligonucleotide Enrichment of HSV-1 Genomic DNA from Clinical Specimens for Use in High-Throughput Sequencing.
To date more than 400 genomes of herpes simplex virus 1 (HSV-1) and the distantly related HSV-2 have been examined using deep sequencing techniques. This powerful approach has been especially useful for revealing the global genetic diversity that exists within and between strains of each virus species. However, most early methods for high-throughput sequencing required the input of abundant viral genomic DNA to enable the successful production of sequencing libraries, and the generation of sufficient short-read sequencing data for de novo genome assembly and similar applications. Therefore, the majority of sequenced HSV strains have been cultured and expanded in vitro prior to genomic analysis, to facilitate isolation of sufficient viral DNA for sequencing-library preparation. Here, we describe an in-solution targeted enrichment procedure for isolating, enriching, and sequencing HSV genomic DNA directly from clinical specimens. When this enrichment technique is combined with traditional sequencing-library preparation procedures, the need for in vitro culturing, expansion, and purification of viral DNA is eliminated. Furthermore, enrichment reduces the large amount of nonviral DNA that is typically present in specimens obtained directly from natural infections, thereby increasing the likelihood of successful viral genome sequencing and assembly. We have used this approach to prepare viral DNA libraries from clinical specimens derived from skin swabs, saliva, blood, and similar sources. We then use these libraries for deep sequencing and successful de novo assembly of the ~152 kb viral genomes, at coverage depths exceeding 100-1000×, for both HSV-1 and HSV-2.


Conjunctival epithelial and goblet cell function in chronic inflammation and ocular allergic inflammation.
Although conjunctival goblet cells are a major cell type in ocular mucosa, their responses during ocular allergy are largely unexplored. This review summarizes the recent findings that provide key insights into the mechanisms by which their function and survival are altered during chronic inflammatory responses, including ocular allergy. Conjunctiva represents a major component of the ocular mucosa that harbors specialized lymphoid tissue. Exposure of mucin-secreting goblet cells to allergic and inflammatory mediators released by the local innate and adaptive immune cells modulates proliferation, secretory function, and cell survival. Allergic mediators like histamine, leukotrienes, and prostaglandins directly stimulate goblet cell mucin secretion and consistently increase goblet cell proliferation. Goblet cell mucin secretion is also detectable in a murine model of allergic conjunctivitis. Additionally, primary goblet cell cultures allow evaluation of various inflammatory cytokines with respect to changes in goblet cell mucin secretion, proliferation, and apoptosis. These findings in combination with the preclinical mouse models help understand the goblet cell responses and their modulation during chronic inflammatory diseases, including ocular allergy. Recent findings related to conjunctival goblet cells provide the basis for novel therapeutic approaches, involving modulation of goblet cell mucin production, to improve treatment of ocular allergies.


[Utero-inguinal hernia].
A ten year old boy was operated for left inguinal hernia at age four years. Male gender was confirmed by sex chromatin. Y corpuscle and male genotype in chromosomal studies, and testicular function was tested by normal testosterone serum levels before and after parenteral human chorionic gonadotropin stimulus. On a second operation, two structurally normal testes and was deferens coexisted with Müllerian structures (rudimentary uterus and two Fallopian tubes of normal histological features) were found. A diagnosis of uterus-inguinalis hernia was made. Deficiencies at Müllerian inhibiting factor's secretion, activity or receptors have been postulated to explain this anomaly.


Characterization of stage-specific antigens of Paragonimus westermani.
In order to develop a serodiagnostic assay to detect antigens of Paragonimus westermani in biological specimens, we generated monoclonal antibodies to antigens of metacercariae and adult worms, and partially characterized the epitopes recognized by representative Mabs. Metacercarial stage-specific determinants were periodate-sensitive and protease-resistant, indicating that they are carbohydrate epitopes. In contrast, adult worm-specific determinants resisted periodate treatment but were sensitive to proteases, indicating that they are polypeptides. The majority of cross-reactive Mabs studied were directed against phosphorylcholine determinants. When used in a dot-ELISA, Paragonimus-specific Mabs could detect 0.3-7 ng/ml parasite antigen in human sera.


Investigation of the status of depression and anxiety among leaders and staff in pharmacy intravenous admixture services in China.
We aimed to investigate the status of depression and anxiety among leaders and staff in pharmacy intravenous admixture services (PIVAS) and analyze influencing factors to provide a reference for improving their mental health status. This study involved a multi-center cross-sectional survey. PIVAS leaders and staff from across China were invited to participate. The Generalized Anxiety Disorder-7 and 9-item Patient Health Questionnaire were used to measure the status of anxiety and depression. A multiple linear regression model was used to analyze influencing factors. In total, 137 PIVAS leaders and 501 staff were included in this study. The results showed that 43.8% (60/137) of the leaders had anxiety and 38.7% (53/137) had depression. Among staff, 47.3% (237/501) had anxiety and 46.5% (233/501) had depression. Multiple linear regression showed that for PIVAS leaders, the degree of recognition by doctors was associated with anxiety scores, whereas PIVAS charge standard, PIVAS profit and loss situation, and the degree of recognition by doctors were associated with depression scores. For PIVAS staff, satisfaction with working in PIVAS was associated with anxiety scores, and job title, working hours per day, and satisfaction with working in PIVAS were associated with depression scores. Depression and anxiety are common among PIVAS leaders and staff working in hospitals in China. Hospitals should implement measures to improve the mental health of PIVAS leaders and staff.


C1q binds phosphatidylserine and likely acts as a multiligand-bridging molecule in apoptotic cell recognition.
Efficient apoptotic cell clearance is critical for maintenance of tissue homeostasis, and to control the immune responses mediated by phagocytes. Little is known about the molecules that contribute "eat me" signals on the apoptotic cell surface. C1q, the recognition unit of the C1 complex of complement, also senses altered structures from self and is a major actor of immune tolerance. HeLa cells were rendered apoptotic by UV-B treatment and a variety of cellular and molecular approaches were used to investigate the nature of the target(s) recognized by C1q. Using surface plasmon resonance, C1q binding was shown to occur at early stages of apoptosis and to involve recognition of a cell membrane component. C1q binding and phosphatidylserine (PS) exposure, as measured by annexin V labeling, proceeded concomitantly, and annexin V inhibited C1q binding in a dose-dependent manner. As shown by cosedimentation, surface plasmon resonance, and x-ray crystallographic analyses, C1q recognized PS specifically and avidly (K(D) = 3.7-7 x 10(-8) M), through multiple interactions between its globular domain and the phosphoserine group of PS. Confocal microscopy revealed that the majority of the C1q molecules were distributed in membrane patches where they colocalized with PS. In summary, PS is one of the C1q ligands on apoptotic cells, and C1q-PS interaction takes place at early stages of apoptosis, in newly organized membrane patches. Given its versatile recognition properties, these data suggest that C1q has the unique ability to sense different markers which collectively would provide strong eat me signals, thereby allowing efficient apoptotic cell removal.


Targeting colon cancer cells with genistein-17.1A immunoconjugate.
We have shown that genistein, a major component of soy, has anti-colon cancer effects in vitro. These effects are attainable at high concentrations that are difficult to achieve in the serum. The purpose of this study was to enhance the activity of genistein against colon cancer cells by coupling it to 17.1A. The monoclonal antibody 17.1A recognizes an epithelial membrane antigen that is overexpressed in colon cancer. Synthesis of Gen-17.1A was achieved by photochemical conjugation using sulfa-SANPAH. Its purity was evaluated by SDS-PAGE. Binding of Gen-17.1A to SW-620 and HT-29 cells was shown using flow cytometry. Internalization was demonstrated by FITC-labeling. Gen-17.1A induced apoptosis in colon cancer cells as evidenced by the acridine orange/ethidium bromide staining method. Gen-17.1A significantly inhibited colon cancer cell growth in vitro and in vivo. Our findings suggest that conjugating genistein to 17.1A monoclonal antibody enhances its effects against colon cancer cells.


[Immunologic aspects of pathologic pain].
The scientific review is devoted to an analysis of neuro-immune aspects of the pathological pain and to the role of disregulation between the central nervous system (CNS) and the immune system in triggering the mechanisms of such pain. The importance of anti-inflammatory cytokines (interleukins, and tumor necrosis factor) as well as of autoantibodies to neuro-mediators in the pathogenesis of different forms of hyperalgetic conditions is evaluated. New data are discussed, which are related with the possibility of modulating the antibodies to neuro-transmitters (serotonin and catecholamines) of experimental neuropathic pain syndromes.


In Situ Electrospun Zein/Thyme Essential Oil-Based Membranes as an Effective Antibacterial Wound Dressing.
Wound dressings are an important element in promoting the healing of wounds. Electrospun fibrous materials have a highly porous structure and controllable antibacterial activity and are therefore popular as potential wound dressings. However, electrospun fibrous wound dressings are usually conveniently packaged for immediate use but cannot accommodate irregularly shaped wounds, and their misuse runs the risk of causing a secondary injury to the wound. To overcome these issues, in situ electrospun zein/thyme essential oil (TEO) nanofibrous membranes are proposed as a potential type of wound dressing and applied for wound management through an in situ electrospinning process, which uses a portable electrospinning device. The as-spun zein/TEO membranes show high gas permeability up to 154 ± 20.9 m2/s and superhydrophilicity with a 0° contact angle. With the addition of TEO, good antibacterial effects are also imparted onto the membrane to prevent infection. Moreover, the in situ electrospinning can directly deposit the zein/TEO membranes onto the site of the wound to accommodate the shape of the wound with increased convenience and perceived comfort. Experiments carried out on mice suggest that the in situ electrospun zein/TEO membrane greatly promotes the wound healing process within 11 days. The study results, therefore, suggest that wound dressings in the form of in situ electrospun zein/TEO membranes can be used to facilitate wound healing.


Tenascin C affects mineralization of SaOS2 osteoblast-like cells through matrix vesicles.
Tenascin C (TNC) is an extracellular matrix glycoprotein involved in osteogenesis and bone mineralization. In a previous study, we identified TNC protein located in the matrix vesicles (MVs) of osteoblasts. MVs are determinant in the mineralization formation. Therefore, we hypothesize whether TNC can modulate osteoblast mineralization via MVs. In this study, we demonstrated that the expression level of TNC was increased with osteoblast differentiation of osteoblast-like SaOS2 cells, and down-regulation of TNC expression by siRNA could significantly inhibit SaOS2 differentiation toward osteoblasts and mineralization as evidenced by decreases in ALP activity, mineralized nodule formation, calcium deposition, and down-regulation of osteogenic marker genes ALP, and COL1A1. Furthermore, we validated that TNC located in the MVs of mineralized SaOS2 cells, and that down-regulation of TNC could decrease MVs mineralization ability in vitro, and the decrease of MVs mineralization ability was not associated with annexins. In conclusion, in this study, we extended the role of TNC during osteogenesis previous progresses, and that supported TNC as an important functional MVs component in modulating osteoblast mineralization.


[Homocystinuria: effectiveness of the treatment with pyridoxine, folic acid, and betaine].
We present the results achieved with vitamin (pyridoxine and folic acid) and betaine (trimethyl-glycine) treatment of three patients with homocystinuria. Cases 1 and 2 were detected by having clinical findings suggestive of the disease (ocular and orthopedic alterations) and case 3 was diagnosed after a family metabolic screening was done. All presented a positive Brand's test and an abnormal elevation of plasma and urine homocysteine, as well as high methionine and low cystine levels in the plasma. Initially, when pyridoxine (600 mg/d) and folic acid (10 mg/d) were given for one month, a partial fall in the homocysteine levels was observed in cases 2 and 3, but not in case 1. When betaine was added (6 g/d), homocysteine disappeared from the plasma after the first month in cases 2 and 3, but only after the third month in case 1. Case 1 also showed a moderate clinical improvement in behavior and school performance. The treatment was maintained for two years in case 1, and for one year in cases 2 and 3. After betaine therapy, no disturbances were observed in the hepatic, renal and bone marrow functions, nor were there any clinically relevant ill-effects. These findings show that betaine offers a therapeutic alternative in the treatment of this disease, independent of the patient's response to pyridoxine.


Plasmapheresis in idiopathic inflammatory myopathy. Experience with 35 patients.
Thirty-five patients with idiopathic inflammatory myopathy (IM) were treated with plasmapheresis. They had had inadequate clinical responses to prednisone or to prednisone and cytotoxic immunosuppressive drug therapy. All patients received cyclophosphamide or chlorambucil in addition to plasmapheresis; 33 also received prednisone. Strength increased in 32 patients during combined therapy; patients with the most active disease experienced substantial improvement that approached clinical remission in some cases. Clinical improvement was documented by muscle testing, forced vital capacity, electromyography, muscle biopsy, and serum creatine phosphokinase activity. The major side effect of treatment was herpes zoster. Primary biliary cirrhosis improved in one patient. Although the effects of plasmapheresis cannot be dissociated from those of the concomitantly administered immunosuppressive drugs, the improvement seen in patients with active disease suggests that plasmapheresis with immunosuppressive drug therapy may find a place in their treatment and implicates circulating factors in the pathogenesis of IM.


The Ca2+-binding sequence in bovine brain S100b protein beta-subunit. A spectroscopic study.
Conformational changes in the beta-subunit of the bovine brain Ca2+-binding protein S100b (S100-beta) accompanying Ca2+ binding were investigated by analysis of the spectroscopic properties of the single tyrosine residue (Tyr17 beta) and flow-dialysis binding experiments. S100-beta binds Ca2+ sequentially at two sites to change the conformation of the protein. The first Ca2+ ion binds to site II beta, a typical Ca2+-binding site in the C-terminal region, and it does not significantly perturb the proximal environment of Tyr17 beta. After the first site is occupied, another Ca2+ ion binds to the N-terminal Ca2+-binding site, I beta, and strengthens a hydrogen bond between Tyr17 beta and a neighbouring carboxylate acceptor group, which results in a large increase in the Tyr17 beta fluorescence spectrum half-width and a positive absorption and c.d. signal between 290 and 275 nm. Ca2+ binding to the S100b.Zn2+6 complex, studied by flow-dialysis and fluorescence measurements showed that, although Zn2+ ions increase the affinity of S100b protein for Ca2+, the Ca2+-binding sequence was not changed. Tb3+ (terbium ion) binding studies on the S100b.Zn2+6 complex proved that Tb3+ antagonizes only Ca2+ binding site II beta and confirmed the sequential occupation of Ca2+-binding sites on the S100b.Zn2+6 complex.


[Molecular basis of the variability of epidemic strains of human influenza viruses].
The data from literature and authors own studies are reviewed on variability of human influenza viral strains, isolated during the same epidemic season in different periods of pandemic cycle. The data obtained indicate that variability of epidemic strains of human influenza virus deals with the genes coding for outer membrane proteins (hemagglutinin and neuraminidase) as well as nonglycosylated proteins. Circulation of a number of viral variants of the same serotype, differing in antigenic specificity of outer membrane proteins or in the genes coding for nonglycosylated proteins was registered during one and the same season of one epidemic. During circulation of viral variants of the same serotype recombination may take place. Heterogeneity of viral strains circulating during different epidemic seasons of the same pandemic cycle is different. The possible mechanisms of development of the new epidemic variants of human influenza virus are discussed.


Understanding the gastrointestinal tract of the elderly to develop dietary solutions that prevent malnutrition.
Although the prevalence of malnutrition in the old age is increasing worldwide a synthetic understanding of the impact of aging on the intake, digestion, and absorption of nutrients is still lacking. This review article aims at filling the gap in knowledge between the functional decline of the aging gastrointestinal tract (GIT) and the consequences of malnutrition on the health status of elderly. Changes in the aging GIT include the mechanical disintegration of food, gastrointestinal motor function, food transit, chemical food digestion, and functionality of the intestinal wall. These alterations progressively decrease the ability of the GIT to provide the aging organism with adequate levels of nutrients, what contributes to the development of malnutrition. Malnutrition, in turn, increases the risks for the development of a range of pathologies associated with most organ systems, in particular the nervous-, muscoskeletal-, cardiovascular-, immune-, and skin systems. In addition to psychological, economics, and societal factors, dietary solutions preventing malnutrition should thus propose dietary guidelines and food products that integrate knowledge on the functionality of the aging GIT and the nutritional status of the elderly. Achieving this goal will request the identification, validation, and correlative analysis of biomarkers of food intake, nutrient bioavailability, and malnutrition.


Spatial and single-cell profiling of the metabolome, transcriptome and epigenome of the aging mouse liver.
Tissues within an organism and even cell types within a tissue can age with different velocities. However, it is unclear whether cells of one type experience different aging trajectories within a tissue depending on their spatial location. Here, we used spatial transcriptomics in combination with single-cell ATAC-seq and RNA-seq, lipidomics and functional assays to address how cells in the male murine liver are affected by age-related changes in the microenvironment. Integration of the datasets revealed zonation-specific and age-related changes in metabolic states, the epigenome and transcriptome. The epigenome changed in a zonation-dependent manner and functionally, periportal hepatocytes were characterized by decreased mitochondrial fitness, whereas pericentral hepatocytes accumulated large lipid droplets. Together, we provide evidence that changing microenvironments within a tissue exert strong influences on their resident cells that can shape epigenetic, metabolic and phenotypic outputs.


Spectral-ripple representation of steady-state vowels in primary auditory cortex.
Responses to various steady-state vowels were recorded in single units in the primary auditory cortex (AI) of the barbiturate-anaesthetized ferret. Six vowels were presented (/a/, /epsilon/, 2 different /i/'s, and 2 different /u/'s) in a natural voiced and a synthetic unvoiced mode. In addition, the responses to broadband stimuli with a sinusoidally shaped spectral envelope (called ripple stimuli) were recorded in each cell, and the response field (RF), which consists of both excitatory and inhibitory regions, was derived from the ripple transfer function. We examined whether the vowel responses could be predicted using a linear ripple analysis method [Shamma et al., Auditory Neurosci. 1, 233-254 (1995)], i.e., by cross correlating the RF of the single unit, and the smoothed spectral envelope of the vowel. We found that for most AI cells (71%) the relative responses to natural vowels could be predicted on the basis of this method. Responses and prediction results for unvoiced and voiced vowels were very similar, suggesting that the spectral fine structure may not play a significant role in the neuron's response to the vowels. Predictions on the basis of the entire RF were significantly better than based solely on best frequency (BF) (or "place"). These findings confirm the ripple analysis method as a valid method to characterize AI responses to broadband sounds as we proposed in a previous paper using synthesized spectra [Shamma and Versnel, Auditory Neurosci. 1, 255-270 (1995)].


Shannon Entropy Loss in Mixed-Radix Conversions.
This paper models a translation for base-2 pseudorandom number generators (PRNGs) to mixed-radix uses such as card shuffling. In particular, we explore a shuffler algorithm that relies on a sequence of uniformly distributed random inputs from a mixed-radix domain to implement a Fisher-Yates shuffle that calls for inputs from a base-2 PRNG. Entropy is lost through this mixed-radix conversion, which is assumed to be surjective mapping from a relatively large domain of size 2J to a set of arbitrary size n. Previous research evaluated the Shannon entropy loss of a similar mapping process, but this previous bound ignored the mixed-radix component of the original formulation, focusing only on a fixed n value. In this paper, we calculate a more precise formula that takes into account a variable target domain radix, n, and further derives a tighter bound on the Shannon entropy loss of the surjective map, while demonstrating monotonicity in a decrease in entropy loss based on increased size J of the source domain 2J. Lastly, this formulation is used to specify the optimal parameters to simulate a card-shuffling algorithm with different test PRNGs, validating a concrete use case with quantifiable deviations from maximal entropy, making it suitable to low-power implementation in a casino.


A scoping review of the Choice and Partnership Approach in child and adolescent mental health services.
Child and adolescent mental health (CAMH) problems are prevalent and inefficient mental health (MH) care systems can contribute to poor outcomes. The Choice and Partnership Approach (CAPA) is a MH care delivery model aiming to provide efficient, high-quality care. Although widely used, no CAPA research review exists. We conducted a scoping review to fill this gap. Medline, Embase, and PsycINFO databases were searched from inception to June 2021. Grey Matters and Google were used to search the grey literature. We identified 5322 records. Removal of duplicates left 4720 documents, which were successively screened and data extracted by pairs of co-authors. The final dataset comprised six published and three non-published studies, conducted primarily at CAMH sites in England, Scotland, Australia, or Canada. Each study had multiple research objectives, which we summarized into seven categories. Positive outcomes were reported for most objectives, but attributing causality to CAPA was hampered by research methodology. Observational designs were used in all studies and approaches to analyzing data varied considerably. Research gaps included the lack of healthcare economics studies of CAPA and no assessment of facilitators and barriers. Current research on CAPA provides intriguing findings meriting further investigation. We suggest strategies to improve future studies.


Repositioning osteotomy for intra-articular malunion of distal radius with radiocarpal and/or distal radioulnar joint subluxation.
Intra-articular malunion of the distal radius may be complicated with radiocarpal and radioulnar joint subluxation, which may result in joint stiffness and loss of function. Conventional corrective osteotomy emphasizes on the restoration of the articular step-off. However, little information is available concerning the restoration of a concentric functioning joint through osteotomy. From 2002 to 2007, 12 patients with chronic intra-articular distal radius fractures were evaluated at an average follow-up of 33.6 months after repositioning osteotomy. The average time from initial injury to reconstructive operation was 11.3 months. The indication for osteotomy included dorsal or volar subluxation of the radiocarpal joint, distal radioulnar joint, or both in addition to articular incongruity. A preoperative computed tomography scan or rapid prototyping (RP) models were performed as part of the surgical planning. Operation was preceded by volar, dorsal, or both approaches. Repositioning osteotomy and internal fixation were also performed. Radiographic analysis and the Disability of Arm, Shoulder and Hand score were used for the outcome assessment. All osteotomy sites healed and all events of radiocarpal and radioulnar subluxation were corrected. The average correction was 13.8 degrees (palmar tilt of the radius) and 1.9 mm in ulnar variance. The mean Disability of Arm, Shoulder and Hand score improved from 64 to 18. Conventional corrective osteotomy via an extra-articular approach was favorably performed to correct an extra-articular malalignment or nascent intra-articular malunion. Problems of abnormal architecture after an intra-articular fracture of the radius are complicated with subluxation of carpus or distal radioulnar joint, which require repositioning via precise articular approach. Both reconstructed computed tomography images and rapid prototyping models are very useful tools in preoperative planning for intra-articular osteotomy. Simulated osteotomy and joint repositioning can be performed in solid models before commencement of actual operation. Repositioning osteotomy consistently restores joint alignment and achieves functional improvement either in cases of nascent simple malunion or complex intra-articular malunion.


Estimating Causal Effects of Treatment in a Randomized Trial When Some Participants Only Partially Adhere.
The intention-to-treat analysis evaluates the causal effect of treatment assignment in a randomized controlled trial; however, participants do not always adhere to assigned treatment and the intention-to-treat effect may differ from the effect of treatment receipt. Although more recent adherence-based methods assess a well-defined causal effect of receiving treatment, adherence is assumed to be dichotomized as all or none. This approach can lack precision in the real world because adherence is a complex and heterogeneous phenomenon. In this article, we illustrate a simple method that provides estimates of bounds on the causal effect of full adherence to treatment in the presence of partial adherence. We first define three types of partial adherence (delayed, partial-dose, posttreatment). We then use casual diagrams to show that categorizing partial adherence as nonadherence in a sensitivity analysis can lead to a violation of the exclusion restriction principle. Finally, we apply recently published sensitivity analyses related to principal stratification that allow for creating bound estimates around the causal effect of treatment in the presence of partial adherence.


Mechanistic characterization of a copper containing thiosemicarbazone with potent antitumor activity.
The thiosemicarbazone CD 02750 (VLX50) was recently reported as a hit compound in a phenotype-based drug screen in primary cultures of patient tumor cells. We synthesized a copper complex of VLX50, denoted VLX60, and characterized its antitumor and mechanistic properties. The cytotoxic effects and mechanistic properties of VLX60 were investigated in monolayer cultures of multiple human cell lines, in tumor cells from patients, in a 3-D spheroid cell culture system and in vivo and were compared with those of VLX50. VLX60 showed ≥ 3-fold higher cytotoxic activity than VLX50 in 2-D cultures and, in contrast to VLX50, retained its activity in the presence of additional iron. VLX60 was effective against non-proliferative spheroids and against tumor xenografts in vivo in a murine model. In contrast to VLX50, gene expression analysis demonstrated that genes associated with oxidative stress were considerably enriched in cells exposed to VLX60 as was induction of reactive oxygen. VLX60 compromised the ubiquitin-proteasome system and was more active in BRAF mutated versus BRAF wild-type colon cancer cells. The cytotoxic effects of the copper thiosemicarbazone VLX60 differ from those of VLX50 and shows interesting features as a potential antitumor drug, notably against BRAF mutated colorectal cancer.


Efficient co-delivery of microRNA 21 inhibitor and doxorubicin to cancer cells using core-shell tecto dendrimers formed via supramolecular host-guest assembly.
Development of versatile and powerful nanoplatforms for efficient therapeutic delivery represents a major topic for current nanomedicine. Herein, we present the development of core-shell tecto dendrimers (CSTDs) for co-delivery of a therapeutic gene and drug for enhanced anticancer therapy applications. In this work, CSTDs were first prepared via supramolecular recognition of β-cyclodextrin (CD)-decorated generation 5 (G5) poly(amidoamine) (PAMAM) dendrimers as cores and adamantane (Ad)-functionalized G3 PAMAM dendrimers as shell components. The formed CSTDs with each G5 dendrimer surrounded with 4.2 G3 dendrimers were evaluated as a gene vector for delivery of plasmid DNA encoding enhanced green fluorescent protein as well as microRNA 21 inhibitor (miR 21i). We show that under an appropriate N/P ratio, the CSTDs enable effective transfection of both genetic materials to cancer cells. In particular, the transfection of miR 21i led to the inhibition of cancer cell migration, decreased miR 21 gene expression, and the effective regulation of the target genes and proteins (e.g., PTEN, PDCD4, p53, and Caspase-3). Furthermore, we revealed that the CSTDs were able to co-deliver miR 21i and an anticancer drug doxorubicin, leading to enhanced therapeutic efficacy to cancer cells in vitro. Our findings imply that the developed CSTDs could be adopted as a versatile platform for effective co-delivery of different therapeutic components for enhanced anticancer therapy applications.


Coordinated Regulation of Anthocyanin Biosynthesis Genes Confers Varied Phenotypic and Spatial-Temporal Anthocyanin Accumulation in Radish (Raphanus sativus L.).
Anthocyanins are natural pigments that have important functions in plant growth and development. Radish taproots are rich in anthocyanins which confer different taproot colors and are potentially beneficial to human health. The crop differentially accumulates anthocyanin during various stages of growth, yet molecular mechanisms underlying this differential anthocyanin accumulation remains unknown. In the present study, transcriptome analysis was used to concisely identify putative genes involved in anthocyanin biosynthesis in radish. Spatial-temporal transcript expressions were then profiled in four color variant radish cultivars. From the total transcript sequences obtained through illumina sequencing, 102 assembled unigenes, and 20 candidate genes were identified to be involved in anthocyanin biosynthesis. Fifteen genomic sequences were isolated and sequenced from radish taproot. The length of these sequences was between 900 and 1,579 bp, and the unigene coverage to all of the corresponding cloned sequences was more than 93%. Gene structure analysis revealed that RsF3'H is intronless and anthocyanin biosynthesis genes (ABGs) bear asymmetrical exons, except RsSAM. Anthocyanin accumulation showed a gradual increase in the leaf of the red radish and the taproot of colored cultivars during development, with a rapid increase at 30 days after sowing (DAS), and the highest content at maturity. Spatial-temporal transcriptional analysis of 14 genes revealed detectable expressions of 12 ABGs in various tissues at different growth levels. The investigation of anthocyanin accumulation and gene expression in four color variant radish cultivars, at different stages of development, indicated that total anthocyanin correlated with transcript levels of ABGs, particularly RsUFGT, RsF3H, RsANS, RsCHS3 and RsF3'H1. Our results suggest that these candidate genes play key roles in phenotypic and spatial-temporal anthocyanin accumulation in radish through coordinated regulation and the major control point in anthocyanin biosynthesis in radish is RsUFGT. The present findings lend invaluable insights into anthocyanin biosynthesis and may facilitate genetic manipulation for enhanced anthocyanin content in radish.


The human hippocampus at 7 T--in vivo MRI.
The human hippocampus plays a central role in various neuropsychiatric disorders, such as temporal lobe epilepsy (TLE), Alzheimer's dementia, mild cognitive impairment, and schizophrenia. Its volume, morphology, inner structure, and function are of scientific and clinical interest. Magnetic resonance (MR) imaging is a widely employed tool in neuroradiological workup regarding changes in brain anatomy, (sub-) volumes, and cerebral function including the hippocampus. Gain in intrinsic MR signal provided by higher field strength scanners and concomitant improvements in spatial resolution seem highly valuable. An examination protocol permitting complete, high-resolution imaging of the human hippocampus at 7 T was implemented. Coronal proton density, T2, T2*, and fluid-attenuated inversion recovery contrasts were acquired as well as an isotropic 3D magnetization-prepared rapid acquisition gradient-echo (500 microm isotropic voxel dimension, noninterpolated). Observance of energy deposition restrictions within acceptable scan times remained challenging in the acquisition of thin, spin-echo-based sections. At the higher resolution enabled by 7 T, demarcation of the hippocampus and some internal features including gray/white matter differentiation and depiction of the hippocampal mantle becomes much more viable when compared with 1.5 T; thus, in the future, this imaging technology might help in the diagnosis of subtle hippocampal changes.


Infrared remote sounding of the middle atmosphere of venus from the pioneer orbiter.
Orbiter infrared measurements of the Venus atmosphere in the 60- to 140-kilometer region show very small diurnal temperature differences near the cloud tops, increasing somewhat at higher levels. The seasonal (that is, equator to pole) contrasts are an order of magnitude larger, and the temperatures unexpectedly increase with increasing latitude below 80 kilometers. An isothermal layer at least two scale heights in vertical extent is found near the 100-kilometer altitude, where the temperature is about 175 K. Structure is present in the cloud temperature maps on a range of spatial scales. The most striking is at high latitude, where contrasts of nearly 50 K are observed between a cold circumpolar band and the region near the pole itself.


Recombination in Enteroviruses, a Multi-Step Modular Evolutionary Process.
RNA recombination is a major driving force in the evolution and genetic architecture shaping of enteroviruses. In particular, intertypic recombination is implicated in the emergence of most pathogenic circulating vaccine-derived polioviruses, which have caused numerous outbreaks of paralytic poliomyelitis worldwide. Recent experimental studies that relied on recombination cellular systems mimicking natural genetic exchanges between enteroviruses provided new insights into the molecular mechanisms of enterovirus recombination and enabled to define a new model of genetic plasticity for enteroviruses. Homologous intertypic recombinant enteroviruses that were observed in nature would be the final products of a multi-step process, during which precursor nonhomologous recombinant genomes are generated through an initial inter-genomic RNA recombination event and can then evolve into a diversity of fitter homologous recombinant genomes over subsequent intra-genomic rearrangements. Moreover, these experimental studies demonstrated that the enterovirus genome could be defined as a combination of genomic modules that can be preferentially exchanged through recombination, and enabled defining the boundaries of these recombination modules. These results provided the first experimental evidence supporting the theoretical model of enterovirus modular evolution previously elaborated from phylogenetic studies of circulating enterovirus strains. This review summarizes our current knowledge regarding the mechanisms of recombination in enteroviruses and presents a new evolutionary process that may apply to other RNA viruses.


The effect of cobalt/copper ions on the structural, thermal, optical, and emission properties of erbium zinc lead borate glasses.
A host glass network of 70B2O3-10Pb3O4-18ZnO-2Er2O3 (ErCoCu1) was proposed and the impact of 1 mol% of Co or/and Cu ions on its structural, thermal, optical, and green emission properties was studied extensively. The X-ray diffraction spectra confirmed the amorphous structure of the produced glasses. Density and density-based parameters behavior showed that the Co or/and Cu ions fill the interstitial positions of the proposed ErCoCu1 network, causing its compactness. Both ATR-FTIR and Raman Spectra affirmed the formation of the fundamental structural units of the borate network, B-O-B linkage, BO3, and BO4. Additionally, the penetration of Co or/and Cu ions inside the ErCoCu1 converts the tetrahedral BO4 units to triangle BO3 causing its richness by non-bridging oxygens. The addition of Co or/and Cu reduces the glass transition temperature as a result of the conversion of the BO4 to BO3 units. Optical absorption spectra for the host glass ErCoCu1 showed many of the distinguished absorption bands of the Er3+ ion. Penetration of Co ion generates two broadbands referring to the presence of Co2+ ions in both tetrahedral and octahedral coordination and Co3+ ions in the tetrahedral coordination. In the Cu-doped glasses, the characteristic absorption bands of Cu2+ and Cu+ were observed. A green emission was generated from the ErCoCu1 glass under 380 nm excitation wavelength. Moreover, no significant effect of Co or/and Cu on the emission spectra was recorded. The considered glasses had appropriate properties qualifying them for optoelectronics and nonlinear optics applications.