Role of peroxisome proliferator-activated receptor-gamma in maintenance of the characteristics of mature 3T3-L1 adipocytes.
Peroxisome proliferator-activated receptor (PPAR)-gamma plays an important role in adipogenesis. However, the functions of PPAR-gamma in differentiated adipocytes have remained unclear. The role of PPAR-gamma in mature 3T3-L1 adipocytes was therefore investigated by overexpression of a dominant negative mutant of this protein (PPAR-gamma-DeltaC) that lacks the 16 COOH-terminal amino acids and that has been shown to prevent the thiazolidinedione-induced differentiation of 3T3-L1 cells into adipocytes. Overexpression of PPAR-gamma-DeltaC in mature 3T3-L1 adipocytes by adenovirus gene transfer resulted in a decrease in both cell size and intracellular triglyceride content, an increase in the extent of lipolysis, and a reduction in the rate of free fatty acid uptake. Furthermore, overexpression of this mutant reduced the abundance of mRNAs for several key enzymes that contribute to triglyceride and free fatty acid metabolism as well as the amounts of GLUT4, insulin receptor, insulin receptor substrate (IRS), and C/EBPalpha mRNAs. It also reduced both the concentration of IRS2 and the insulin-stimulated glucose uptake. These results suggest that PPAR-gamma plays an important role in mature 3T3-L1 adipocytes at least in part by maintaining the expression of genes that confer the characteristics of mature adipocytes.


Meta-analysis of occupational exposures in the rubber manufacturing industry and risk of cancer.
Occupational exposures in the rubber manufacturing industry showed an increased risk of cancer and have been classified as a group 1 carcinogen, largely from studies on workers employed before 1950s. Cancer sites considered as causally associated are bladder, lung and stomach, and leukaemia. Recent studies did not report an increased risk of cancer. A meta-analysis was conducted on observational studies published until April 2016 on occupational exposures in the rubber manufacturing industry and risk of cancer. Case-control and cohort studies were included. Random effect models were used. Heterogeneity and publication bias were evaluated. Stratified analyses were conducted on study characteristics. The literature search identified 46 cohorts and 59 case-control studies. An increased risk was found for bladder cancer [standardised incidence ratio (SRR) = 1.36; 95% confidence interval (CI) 1.18, 1.57], leukaemia (SRR = 1.29; 95% CI 1.11, 1.52), lymphatic and haematopoietic system (SRR = 1.16; 95% CI 1.02, 1.31) and larynx cancer (SRR = 1.46; 95% CI 1.10, 1.94). For lung cancer, a borderline statistically significant increased risk was identified (SRR = 1.08; 95% CI 0.99, 1.17). No association was found for stomach cancer (SRR = 1.06; 95% CI 0.95, 1.17). In stratified analyses, risks of cancer were not increased for workers employed after 1960 for bladder cancer (SRR = 1.06; 95% CI 0.66, 1.71), lung cancer (SRR = 0.94; 95% CI 0.68, 1.29) or leukaemia (SRR = 0.92; 95% CI 0.62, 1.36). Risk of bladder cancer, lung cancer, leukaemia and larynx cancer were increased among workers in rubber industry. Evidence of elevated risks was no longer seen for bladder cancer, lung cancer or leukemia among workers first employed after 1960.


Solid phase click ligation for the synthesis of very long oligonucleotides.
Oligonucleotides have been ligated efficiently on solid-phase using CuAAC and SPAAC chemistry to produce up to 186-mer triazole linked DNA products. Multiple sequential ligation reactions can be carried out by using a masked azide approach. This work suggests a novel modular approach to the synthesis of large complex oligonucleotide analogues.


A new mulinane diterpenoid from the cushion shrub Azorella compacta growing in Perú.
Azorella compacta is a rare yellow-green compact resinous cushion shrub growing from the high Andes of southern Perú to northwestern Argentina, and which is a producer of biologically active and unique diterpenoids. This study investigated the secondary metabolites present in a Peruvian sample of Azorella compacta and the evaluation of gastroprotective activity of the isolated compounds in a gastric- induced ulcer model in mice. Six secondary metabolites (diterpenoids 1-6) present in the dichloromethane (DCM) extract of A. compacta growing in Perú were isolated by a combination of Sephadex LH-20 permeation and silica gel chromatography and their chemical structures were elucidated by spectroscopic methods (NMR) and molecular modeling. The gastroprotective activity of the new compound 1 was evaluated on the HCl/EtOH-induced gastric lesion model in mice and compared to the activity showed by the known compounds. A new mulinane diterpene along with five known diterpenoids have been isolated from a Peruvian sample of A. compacta and the gastroprotective results show that compound 1 is less active than the other known mulinane diterpenoids isolated. A. compacta growing in Perú showed the presence of the new mulinane 1, which was poorly active in the HCl/EtOH-induced gastric lesion model in mice. Indeed, the activity was lower than other diterpenoids (2-6) showing an oxygenated function at C-16 or/and C-20, which confirm the role of an oxygenated group (OH or carboxylic acid) for the gastroprotective activity of mulinane compounds.


Prostate-specific antigen-retargeted recombinant newcastle disease virus for prostate cancer virotherapy.
Oncolytic virus (OV) therapies of cancer are based on the use of replication-competent, tumor-selective viruses with limited toxicity. Newcastle disease virus (NDV), an avian paramyxovirus, is a promising OV and is inherently tumor selective and cytotoxic only to tumor cells. Replication is restricted in normal cells. Despite encouraging phase I/II clinical trials with NDV, further refinements for tumor-specific targeting are needed to enhance its therapeutic index. Systemically delivered NDV fails to reach solid tumors in therapeutic concentrations and also spreads poorly within the tumors due to barriers including complement, innate immunity, and the extracellular matrix. Overcoming these hurdles is paramount to realizing the exceptional oncolytic efficacy of NDV. We engineered the F protein of NDV and generated a recombinant NDV (rNDV) whose F protein is cleavable exclusively by prostate-specific antigen (PSA). The rNDV replicated efficiently and specifically in prostate cancer (CaP) cells and 3-dimensional prostaspheres but failed to replicate in the absence of PSA. Induction of intracellular PSA production by a synthetic androgen analog (R1881) enhanced fusogenicity in androgen-responsive CaP cells. Further, PSA-cleavable rNDV caused specific lysis of androgen-independent and androgen-responsive/nonresponsive CaP cells and prostaspheres, with a half-maximal effective concentration (EC50) ranging from a multiplicity of infection of 0.01 to 0.1. PSA-retargeted NDV efficiently lysed prostasphere tumor mimics, suggesting efficacy in vivo. Also, PSA-cleavable NDV failed to replicate in chicken embryos, indicating no pathogenicity for chickens. Prostate-specific antigen targeting is likely to enhance the therapeutic index of rNDV owing to tumor-restricted replication and enhanced fusogenicity.


Conductive Na+ transport in an epithelial cell line (LLC-PK1) with characteristics of proximal tubular cells.
Na+ influx and efflux from confluent monolayers of an epithelial cell line with multiple differentiated characteristics of the straight segment of the renal proximal tubule were studied in the presence and absence of a pH gradient. The results show that Na+ influx in the absence of a pH gradient is inhibited by amiloride as well as by complete replacement of Cl- by an impermeable anion, such as isethionate. Dissipation of cell membrane potential by increasing the potassium concentration of the extracellular medium in the presence of valinomycin also inhibited Na+ influx, whereas sodium influx induced by an H+ gradient was not affected. Inhibition of Na+ influx by different maneuvers produced hyperpolarization of the plasma cell membrane, as would be expected if the sodium movement involved net displacement of charges. Calcium and other divalent and trivalent cations also inhibited Na+ influx measured in the absence of an H+ gradient. Na+ influx induced by a pH gradient, however, was not affected. Like the Na+-H+-exchange system, the conductive Na+ pathway is localized in the apical membrane of the epithelial cells. From these results, we conclude that at least a fraction of transepithelial Na+ transport in LLC-PK1 monolayers occurs through a simple rheogenic transport system.


Efficient Delivery of Quantum Dots into the Cytosol of Cells Using Cell-Penetrating Poly(disulfide)s.
Quantum dots (QDs) are extremely bright, photostable, nanometer particles broadly used to investigate single molecule dynamics in vitro. However, the use of QDs in vivo to investigate single molecule dynamics is impaired by the absence of an efficient way to chemically deliver them into the cytosol of cells. Indeed, current methods (using cell-penetrating peptides for instance) provide very low yields: QDs stay at the plasma membrane or are trapped in endosomes. Here, we introduce a technology based on cell-penetrating poly(disulfide)s that solves this problem: we deliver about 70 QDs per cell, and 90% appear to freely diffuse in the cytosol. Furthermore, these QDs can be functionalized, carrying GFP or anti-GFP nanobodies for instance. Our technology thus paves the way toward single molecule imaging in cells and living animals, allowing to probe biophysical properties of the cytosol.


Wireless Galvanic Impulse Communication for High-Throughput, Low-Power, Miniaturized Neuromodulation Implants.
Deeply implanted bioelectronic devices that selectively record and stimulate peripheral nerves have the potential to revolutionize healthcare by delivering on-demand, personalized therapy. A key barrier to this goal is the lack of a miniaturized, robust, and energy-efficient wireless link capable of transmitting data from multiple sensing channels. To address this issue, we present a wireless galvanic impulse link that uses two 500μm diameter planar electrodes on the outside of a nerve cuff to transmit data to a wearable receiver on the skin's surface at rates greater than 1Mbps. To achieve an energy-efficient, high data rate link, our protocol encodes information in the timing of narrow biphasic pulses that is reconstructed by the wearable receiver. We use a combination of modeling and in vivo and in vitro experimentation to demonstrate the viability of the link. We demonstrate losses lower than 60dB even with significant, 50mm lateral misalignment, ensuring a sufficient signal-to-noise ratio for robust operation. Using a custom, flexible nerve cuff, we demonstrate data transmission in a 14mm-thick rodent animal model and in a 42mm-thick heterogeneous human tissue phantom.


Causality, consciousness, and cerebral organization.
This article is based upon data which are suitable for the correlation of behavioral research and experimental neurophysiology. Causal thinking manifests a sort of integrative activity which brings simultaneous and successive patterns of nervous excitation into a subjectively meaningful frame of reference. While neuronal patterns determine the content of consciousness, they fail to provide clues concerning the transformation of such patterns into subjective experience.


Randomly inserted and targeted Hox/reporter fusions transcriptionally silenced in Polycomb mutants.
Polycomb-group (Pc-G) proteins ensure late maintenance of transcriptional repression outside the expression domain of target genes in flies and vertebrates. They act in complexes, presumably by modulating chromatin structure. In Drosophila, they have been found to be associated with transcriptionally inactive loci but seem to be present in association with actively transcribed promoters as well, a feature which is not yet understood. In the mouse, mutations in several Pc-G genes result in an often subtle, local derepression of only a subset of the Hox genes rostral to their expression domains. We report here that Hox/reporter fusion genes, either randomly integrated as transgenes or as insertions within endogenous loci, are transcriptionally silenced in two mouse Pc-G-null mutants, Mel18 and rae28. Transcriptional silencing of Hox/reporter transgenes in Pc-G mutants was accompanied by increased DNA methylation in the promoter region. Gene silencing was observed at early developmental stages, long before Pc-G and trithorax-group proteins exert their function in maintenance of the Hox patterns. Although all five Hox genes tested as Hox/reporter fusions were silenced in the Pc-G mutants, transcription of the endogenous loci was mildly decreased in a subset of these Hox genes, and Hoxb1 was the most strongly affected. We discuss the possibilities that the observed negative effect of Pc-G mutations on Hox and Hox/reporter expression may reflect a positive involvement of the Pc-G epigenetic repressors in initial Hox gene transcription and that this requirement is exacerbated by the reporter insertion.


Assessment of Na+/K+ ATPase Activity in Small Rodent and Human Skeletal Muscle Samples.
In skeletal muscle, the Na/K ATPase (NKA) plays essential roles in processes linked to muscle contraction, fatigue, and energy metabolism; however, very little information exists regarding the regulation of NKA activity. The scarcity of information regarding NKA function in skeletal muscle likely stems from methodological constraints, as NKA contributes minimally to total cellular ATP utilization, and therefore contamination from other ATPases prevents the assessment of NKA activity in muscle homogenates. Here we introduce a method that improves accuracy and feasibility for the determination of NKA activity in small rodent muscle samples (5-10 mg) and in human skeletal muscle. Skeletal muscle homogenates from mice (n = 6) and humans (n = 3) were used to measure NKA and sarcoplasmic reticulum Ca ATPase (SERCA) activities with the addition of specific ATPase inhibitors to minimize "background noise." We observed that myosin ATPase activity was the major interfering factor for estimation of NKA activity in skeletal muscle homogenates, as the addition of 25 μM of blebbistatin, a specific myosin ATPase inhibitor, considerably minimized "background noise" (threefold) and enabled the determination of NKA maximal activity with values three times higher than previously reported. The specificity of the assay was demonstrated after the addition of 2 mM ouabain, which completely inhibited NKA. On the other hand, the addition of blebbistatin did not affect the ability to measure SERCA function. The coefficient of variation for NKA and SERCA assays were 6.2% and 4.4%, respectively. The present study has improved the methodology to determine NKA activity. We further show the feasibility of measuring NKA and SERCA activities from a common muscle homogenate. This methodology is expected to aid in our long-term understanding of how NKA affects skeletal muscle metabolic homeostasis and contractile function in diverse situations.


Mouse c1q: light and electron microscopic immunohistochemical localization.
The localization of mouse C1q in various mouse tissues was assessed by immunohistochemistry. In all mouse strains examined, C1q was associated with cells in small focal areas of lymph nodes, Peyer's patches, and spleens often near germinal centers. Ultrastructural studies of these foci demonstrated that C1q was present in the endoplasmic reticulum of mononucleated cells that had an extensive cytoplasmic network. Occasionally, small deposits of C1q were observed in the perinuclear space of mononucleated cells that had scanty cytoplasm with few organelles. Extracellular C1q was found at the surface of interdigitating cell processes in these lymphatic foci. In addition to the presence of C1q within lympho-reticular tissues, C1q frequently was detected in renal glomeruli but not in urinary tract epithelium or intestinal epithelium of adult mice; in contrast, C1q was localized in the intestinal epithelium of neonatal mice. C1q was not detected in tissue fibroblasts nor in cultured fibroblasts; however, a subpopulation of mouse adherent peritoneal cells maintained in vitro did contain C1q. The results of these studies provide new information regarding the distribution and, thus, the possible sites of C1q biosynthesis in normal mouse tissues.


The nuclear envelope in the crystalline lens fiber cell.
Rabbit lenses which have been fixed, dehydrated, and dried by a critical-point drying method, can be fractured through the cytoplasm of the differentiating lens fibers, exposing the cell nuclei. The fracture, under these conditions, causes a complete separation of the two membranes of the nuclear envelope from one another, thus exposing entire membrane surfaces (those which line the perinuclear space). These surfaces are not seen in their entirety in typical freeze-fracture or freeze-etch preparations, and consequently have not been described previously. The exposed membrane surfaces which line the perinuclear space have numerous convex structures of approximately 1,000 A, and some larger more irregularly shaped structures. These appear to be fragments of the nuclear pore complexes. Differences in these structures between young fibers and those nearing completion of differentiation is suggested.


Photocontact allergy to benzocaine.
A photodermatitis developed in two patients after the use of commercial sunscreens and a topical anesthetic lotion containing benzocaine. Photopatch testing indicated the presence of photocontact allergy to benzocaine. One patient had a positive photopatch test to a commercial glyceryl para-aminobenzoic acid (PABA) preparation that was heavily contaminated with benzocaine, but not to a benzocaine-free sample of glyceryl PABA. Benzocaine failed to produce phototoxic reactions in normal volunteers. Efforts to induce photocontact allergy in guinea pigs were unsuccessful.


Bullous dermatosis of hemodialysis.
Bullous dermatosis of hemodialysis (BDH), first described in 1975, is characterized by porphyrialike blisters on sun-exposed parts of the scalp, face, arms, and hands. Four patients with BDH are presented. In two of these patients the deliberate lowering of their serum iron levels led to improvement of the condition. Further investigation into the role of iron metabolism in BDH is suggested.


Prevalence and predictors of unknown HIV status among women delivering in Mulago National Referral Hospital, Kampala, Uganda.
Knowledge of a person's HIV status during pregnancy is critical for prevention of mother to child transmission of HIV. To determine the prevalence and factors associated with unknown HIV status among women delivering in Mulago Hospital. This was a cross-sectional study of women that had just delivered. The women's demographic characteristics, health seeking behaviour, health system-related factors and knowledge on PMTCT were collected. Fisher's exact test, Wilcoxon rank sum test and logistic regression were used to test associations. The prevalence of unknown HIV status was 2.6% (10/382). Attending ANC at higher level facilities (OR =0.1 95% CI 0.0 - 0.4) and having been counselled for HIV testing during ANC (OR=0.1, 95% CI 0.0 - 0.4) were associated with likelihood of having a known HIV status. Out of the ten women with unknown HIV status, 4/6 who attended ANC in public/government accredited health facilities "opted out" of HIV testing due to personal reasons. Among the four who attended ANC in private clinics, two were not offered HIV testing and one "opted out". Most participants had a known HIV status at labour (97%). Private clinics need to be supported to provide free quality HCT services in ANC.


Uptake and predictors of contraceptive use in Afghan women.
Afghanistan has one of the world's highest fertility rates and, related to this, an infant mortality rate far higher than its South Asian neighbors. Contraception enhances family spacing, improves women's safety in child birth and, as a result, reduces infant and child mortality. Until recently, there has been a paucity of information on the comparative rates of contraceptive practices in the country and socioeconomic correlates of uptake. We aimed to elucidate the factors influencing the use of contraception in Afghanistan using recent, robust national data. Using Afghanistan Mortality Survey (AMS) 2010 data, the distribution of Contraceptive Prevalence Rate (CPR) and correlates of contraceptive use among currently married women aged 15-49 years were explored. We initially summarised descriptive data on 25,743 married women and then derived predictors of the use of any form of contraception using a multiple logistic regression model. The prevalence of self-reported current use of any contraceptive method was 21.8% (95% CI: 20.4-23.4) at the national level though there was a wide variation in practice between provinces. Herat province in the West region had a highest contraceptive prevalence rate of 49.4% while Paktika in the Southeast region had the lowest CPR of 2%. Multiple logistic regression analysis showed that a family size of greater than 6 living children strongly predicted contraceptive use (AOR 7.4 (95% CI:6.1-9.0)). Other independent predictors included: secondary or high level of education (AOR 2.1 (95% CI: 1.8-2.5)) and being in the wealthiest stratum (OR 2.1 (95% CI 1.5-3.0)). Rural residence predicted a lower use of contraception (AOR, 0.72; 95% CI: 0.56-0.92). Contraceptive uptake rate was low overall with wide inter provincial variation. Strengthening female education, targeting married women in rural area and women with no education may enhance the effectiveness of National Family planning program in Afghanistan.


Quasi-BIC Mode Lasing in a Quadrumer Plasmonic Lattice.
Plasmonic lattices of metal nanoparticles have emerged as an effective platform for strong light-matter coupling, lasing, and Bose-Einstein condensation. However, the full potential of complex unit cell structures has not been exploited. On the other hand, bound states in continuum (BICs) have attracted attention, as they provide topologically protected optical modes with diverging quality factors. Here, we show that quadrumer nanoparticle lattices enable lasing in a quasi-BIC mode with a highly out-of-plane character. By combining theory with polarization-resolved measurements of the emission, we show that the lasing mode has a topological charge. Our analysis reveals that the mode is primarily polarized out-of-plane as a result of the quadrumer structure. The quality factors of the out-of-plane BIC modes of the quadrumer array can be exceedingly high. Our results unveil the power of complex multiparticle unit cells in creating topologically protected high-Q modes in periodic nanostructures.


The human chondrosarcoma HCS-2/8 cell line is responsive to BMP-7, but not to IL-1beta.
Cultures of primary chondrocytes as in vitro model systems for studying the cellular behavior of chondrocytes are notoriously difficult to cultivate and propagate. One way to circumvent these problems appears to be the use of immortalized/immortal chondrocytic cell lines. In the present study, we were interested whether the chondrosarcoma derived HCS-2/8 cells are suitable for studying major cellular reaction pattern in response to key anabolic (BMP-7) and catabolic (IL-1beta) factors. Therefore, we used cDNA array and real-time PCR technology in order to evaluate gene expression triggered by stimulation with IL-1beta (0,1-100 ng/ml) and BMP-7 in confluent monolayer cultures. HCS-2/8 cells hardly responded to IL-1beta, but showed good responsiveness to BMP-7. We found 12 genes up- and 17 significantly down-regulated by BMP-7 (out of 340 investigated genes). Besides the expected activation of anabolic genes chondrocytic cells after BMP-stimulation try to neutralize activation of the BMP-signalling cascade by expressing intra- and extracellular BMP-antagonists. Chondrosarcoma derived cell lines are a potential substitute for primary articular chondrocytes promising consistent expression of a differentiated chondrocyte phenotype with sufficient proliferative capacity. However, as shown by this study one needs to carefully select the cell line depending on the effects which one intends to study. In this respect, HCS-2/8 cells are a validated tool for studying BMP-effects on chondrocytes, but not e.g. effects of interleukin-1.


[The mucosal electrosensitivity of the anal canal following restorative proctocolectomy for ulcerative colitis].
The aim of this study was to investigate the mucosal electrosensitivity of the anus in 23 patients following restorative proctocolectomy for ulcerative colitis. In 16 cases mechanical anastomosis was performed 2 cm above the pectineal line conserving the transitional anal mucosa and in 7 cases distal rectal mucosectomy and manual anastomosis was performed to the pectineal line. In these patients and in those of the control group of 21 patients without previous anorectal disease, the threshold of sensitivity of the lower, middle and upper anus segment was determined. A clinical questionnaire was made in the patients with ileoanal reservoir to evaluate the existence of alterations in continence, and in the capacity to discriminate between gases and stools. The sensitivity thresholds obtained were significantly higher than those in the control group. A significantly greater sensitivity threshold (p = 0.01) was found in the upper anus segment in patients with mucosectomy (median 18.5 mA) with respect to the patients with mechanical anastomosis (median 11 mA). A significantly greater threshold was also observed (p = 0.03) globally in the patients with some alteration in continence (median 15 mA; 57.1% mucosectomy and 18.8% mechanical anastomosis) with respect to completely continent patients (median 11 mA). The results demonstrate the importance of conserving the transitional anal mucosa during surgery with the aim of improving the functional results during the follow of the functioning ileoanal reservoir.


The cannabinoid receptor antagonist SR 141716 attenuates overfeeding induced by systemic or intracranial morphine.
Considerable interplay exists between the brain's opioid and cannabinoid systems. These systems are both involved in the control of appetite and research supports the notion that the opioid system modulates the role of the cannabinoid system on appetite. However, the ability of the cannabinoid system to modulate the opioid system's control over appetite has not been well studied. The present study examined the role of cannabinoid CB(1) receptors in the control of opioid-induced feeding, and sought to identify specific brain regions underlying this role. After being habituated to the test environment and injection procedure, sated rats were injected with the cannabinoid CB(1) receptor antagonist SR 141716 (0.03-3.0 mg/kg, IP). Thirty minutes later, morphine or its vehicle were administered systemically (2.5 mg/kg SC, experiments 1 and 2) or intracranially into the nucleus accumbens (nAcc, experiment 3) or paraventricular nucleus of the hypothalamus (PVN, experiment 4). Food intake and locomotor activity was then recorded for 120 min. A significant increase in food intake was observed following systemic and intracranial (10 nmol) application of morphine in all experiments. SR 141716 suppressed systemic and intra-PVN morphine induced feeding (experiments 2 and 4), but did not attenuate food intake induced by intra-nAcc application of morphine (experiment 3). Because SR 141716 had no effect on intra-nAcc morphine-stimulated feeding, it would appear that cannabinoid receptors do not modify opioid-mediated hedonic responses to food. Rather, we conclude that cannabinoid CB(1) receptor blockade may suppress opioid-induced feeding by stimulating the release of satiety-related peptides within the hypothalamus. Further, because SR 141716 did not block morphine induced locomotor activity, the observed effects on feeding do not appear to be due to a non-specific reduction in motivated behaviour.


Field bioefficacy of deltamethrin residual spraying against dengue vectors.
Field bioefficacy of residual-sprayed deltamethrin against Aedes vectors was evaluated in an urban residential area in Kuala Lumpur. The trial area consisted of single storey wood-brick houses and a block of flat. The houses were treated with outdoor residual spraying while the flat was used as an untreated control. Initial pre-survey using ovitrap surveillance indicated high Aedes population in the area. Deltamethrin WG was sprayed at a dosage of 25mg/m2 using a compression sprayer. The effectiveness of deltamethrin was determined by wall bioassay and ovitrap surveillance. The residual activity of 25mg/m2 deltamethrin was still effective for 6 weeks after treatment, based on biweekly bioassay results. Bioassay also indicated that both Aedes aegypti and Aedes albopictus were more susceptible on the wooden surfaces than on brick. Aedes aegypti was more susceptible than Ae. albopictus against deltamethrin. Residual spraying of deltamethrin was not very effective against Aedes in this study since the Aedes population in the study area did not reduce as indicated by the total number of larvae collected using the ovitrap (Wilcoxon Sign Test, p> 0.05). Further studies are required to improve the effectiveness of residual spraying against Aedes vectors.


[Effect of selenium enriched diet on lipid peroxidation in patients with diabetes mellitus type 2].
It was investigated the influence of a diet supplemented with selenium on the clinicometabolic parameters and the activity of lipid peroxidation in 43 patients with Type 2 diabetes mellitus. It was indicated that under the influence of dietotherapy besides the decrease of glucose levels in capillary and venous blood, improvement of parameters of lipid metabolism, a reduction of malondialdehyde level and tendency to increase of glutathione peroxidase activity were observed. The results of investigations testify about sufficiently expressed efficacy of a diet supplemented with selenium in correction of oxidative stress and clinicometabolic parameters in this contingent of patients.


Speech and oro-motor function in children with developmental coordination disorder: a pilot study.
The protracted maturation and development of speech articulation underlies the complexity of the skill, and suggests it may be an area susceptible to a general deficit in motor control. Recent research suggests a high co-occurrence between Developmental Coordination Disorder (DCD) and disordered speech production. Despite this there has been no systematic investigation of speech motor control in children with DCD. We conducted a pilot study which looked at speech motor control in a group of children with DCD (N=5) and a group typically developing (TD) children (N=5). Movements of the upper and lower lip were recorded during non-verbal movements, single words, syllable sequences, and sentence repetition. In the baseline conditions (normal talking speed or an isolated utterance) children with DCD demonstrated a typical pattern of movement, albeit a slower and shorter movement. In contrast, when task complexity was increased the children with DCD showed an atypical pattern of movement. It was concluded that children with DCD demonstrate inferior motor control for complex speech gestures, suggesting that the motor deficit in DCD may indeed be a more generalized phenomenon affecting the speech motor system.


New bio-ceramization processes applied to vegetable hierarchical structures for bone regeneration: an experimental model in sheep.
Bone loss is still a major problem in orthopedics. The purpose of this experimental study is to evaluate the safety and regenerative potential of a new scaffold based on a bio-ceramization process for bone regeneration in long diaphyseal defects in a sheep model. The scaffold was obtained by transformation of wood pieces into porous biomorphic silicon carbide (BioSiC®). The process enabled the maintenance of the original wood microstructure, thus exhibiting hierarchically organized porosity and high mechanical strength. To improve cell adhesion and osseointegration, the external surface of the hollow cylinder was made more bioactive by electrodeposition of a uniform layer of collagen fibers that were mineralized with biomimetic hydroxyapatite, whereas the internal part was filled with a bio-hybrid HA/collagen composite. The final scaffold was then implanted in the metatarsus of 15 crossbred (Merinos-Sarda) adult sheep, divided into 3 groups: scaffold alone, scaffold with platelet-rich plasma (PRP) augmentation, and scaffold with bone marrow stromal cells (BMSCs) added during implantation. Radiological analysis was performed at 4, 8, 12 weeks, and 4 months, when animals were sacrificed for the final radiological, histological, and histomorphometric evaluation. In all tested treatments, these analyses highlighted the presence of newly formed bone at the bone scaffolds' interface. Although a lack of substantial effect of PRP was demonstrated, the scaffold+BMSC augmentation showed the highest value of bone-to-implant contact and new bone growth inside the scaffold. The findings of this study suggest the potential of bio-ceramization processes applied to vegetable hierarchical structures for the production of wood-derived bone scaffolds, and document a suitable augmentation procedure in enhancing bone regeneration, particularly when combined with BMSCs.


Inhibitory effects of ZSTK474, a novel phosphoinositide 3-kinase inhibitor, on osteoclasts and collagen-induced arthritis in mice.
Targeting joint destruction induced by osteoclasts (OCs) is critical for management of patients with rheumatoid arthritis (RA). Since phosphoinositide 3-kinase (PI3-K) plays a critical role in osteoclastogenesis and bone resorption, we examined the effects of ZSTK474, a novel phosphoinositide 3-kinase (PI3-K)-specific inhibitor, on murine OCs in vitro and in vivo. The inhibitory effect of ZSTK474 on OC formation was determined and compared with other PI3-K inhibitors by counting tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells after culturing murine bone marrow monocytic OC precursors, and RAW264.7 cells. Activation of Akt and expression of nuclear factor of activated T cells (NFAT) c1 in cultured RAW264.7 cells were examined. The suppressing effect of ZSTK474 on bone resorption was assessed by the pit formation assay. The in vivo effects of ZSTK474 were studied in collagen-induced arthritis (CIA) in the mouse. Oral daily administration of ZSTK474 was started either when more than half or when all mice developed arthritis. Effects of ZSTK474 were evaluated using the arthritis score and histological score of the hind paws. ZSTK474 inhibited the differentiation of bone marrow OC precursors and RAW264.7 cells in a dose-dependent manner. The inhibitory effect of ZSTK474 was much stronger than that of LY294002, the most commonly used PI3-K inhibitor. In addition, ZSTK474 suppressed the bone resorbing activity of mature OCs. Moreover, oral daily administration of ZSTK474, even when begun after the development of arthritis, ameliorated CIA in mice without apparent toxicity. Histological examination of the hind paw demonstrated noticeable reduction of inflammation and of cartilage destruction in ZSTK474-treated mice. ZSTK474 also significantly decreased OC formation adjacent to the tarsal bone of the hind paw. These findings suggest that inhibition of PI3-K with ZSTK474 may potentially suppress synovial inflammation and bone destruction in patients with RA.


Top Ten Tips Palliative Care Clinicians Should Know About Integrative Palliative Care.
Integrative medicine (IM) use is widespread among individuals living with serious illness. There is a natural alignment between the fields of IM and palliative care (PC) rooted in their shared core values. Integrative palliative care (IPC) is an emerging focus within the field of PC that aims to broaden the healing toolkit available to patients with serious illness by combining standard-of-care biomedical treatments with evidence-informed integrative and complementary medicine practices with the goal of enhancing quality of life at every stage of a person's health journey. This article is an evidence-based guide to incorporating IPC practices into the care of seriously ill individuals.


SMA-miRs (miR-181a-5p, -324-5p, and -451a) are overexpressed in spinal muscular atrophy skeletal muscle and serum samples.
Spinal muscular atrophy (SMA) is a neuromuscular disorder characterized by the degeneration of the second motor neuron. The phenotype ranges from very severe to very mild forms. All patients have the homozygous loss of the SMN1 gene and a variable number of SMN2 (generally 2-4 copies), inversely related to the severity. The amazing results of the available treatments have made compelling the need of prognostic biomarkers to predict the progression trajectories of patients. Besides the SMN2 products, few other biomarkers have been evaluated so far, including some miRs. We performed whole miRNome analysis of muscle samples of patients and controls (14 biopsies and 9 cultures). The levels of muscle differentially expressed miRs were evaluated in serum samples (51 patients and 37 controls) and integrated with SMN2 copies, SMN2 full-length transcript levels in blood and age (SMA-score). Over 100 miRs were differentially expressed in SMA muscle; 3 of them (hsa-miR-181a-5p, -324-5p, -451a; SMA-miRs) were significantly upregulated in the serum of patients. The severity predicted by the SMA-score was related to that of the clinical classification at a correlation coefficient of 0.87 (p<10-5). miRNome analyses suggest the primary involvement of skeletal muscle in SMA pathogenesis. The SMA-miRs are likely actively released in the blood flow; their function and target cells require to be elucidated. The accuracy of the SMA-score needs to be verified in replicative studies: if confirmed, its use could be crucial for the routine prognostic assessment, also in presymptomatic patients. Telethon Italia (grant #GGP12116).


[Variability of seizure phenomenology in the light of stereo-EEG studies].
The demonstration of a largely stable electroclinical semiology in the early phase of seizures is a main prerequisite for successful causative surgery of epilepsy. A few experiences with analysis of similarities during seizures are presented and discussed regarding their fundamental pathogenetic but also practical clinical significance. The primary data stem from candidates for selective surgery of the temporal lobe, mainly afflicted by psychomotor seizures. These seizures were recorded either by invasive stereoelectroencephalography or by semi-invasive foramen ovale electrodes.


Power and challenges of using zebrafish as a model for skeletal tissue imaging.
The zebrafish (Danio rerio) is now a widely used model organism in biomedical research. The species is also increasingly used for studying skeletal development and regeneration and for understanding human skeletal diseases. The small size of this model organism is an advantage and an extreme challenge for visualizing and diagnosing the animals' skeleton. This applies especially to early stages of skeletal development. Similar challenges arise for the analysis of the skeleton of other small fish species, such as medaka (Oryzias latipes). High quality histological preparations and knowledge about the special quality of the zebrafish skeleton remain prerequisites for a correct analysis. In addition, new methods for fast and high-resolution 2D and 3D skeletal tissue screening are required for a maximal understanding of skeletal development. We, in this study, review advantages and limitations of adapting current visualization techniques for zebrafish skeletal research. We discuss the methods for in toto visualization, such as X-raying, micro-CT, Alizarin red staining and optical projection tomography. Techniques for in vivo imaging, such as second harmonic generation microscopy and two-photon excitation fluorescence, are also discussed. Finally, we explore the possibilities of light-sheet microscopy for the analysis of the zebrafish skeleton.


Short and Long term predictions of Hospital emergency department attendances.
Emergency departments in the United Kingdom (UK) experience significant difficulties in achieving the 95% NHS access standard due to unforeseen variations in patient flow. In order to maximize efficiency and minimize clinical risk, better forecasting of patient demand is necessary. The objective is therefore to create a tool that accurately predicts attendance at emergency departments to support optimal planning of human and physical resources. Historical attendance data between Jan-2011 - December-2015 from four hospitals were used as a training set to develop and validate a forecasting model. To handle weekday variations, the data was first segmented into each weekday time series and a separate model for each weekday was performed. Seasonality testing was performed, followed by Box-Cox transformations. A modified heuristics based on a fuzzy time series model was then developed and compared with autoregressive integrated moving average and neural networks models using Harvey, Leybourne and Newbold (HLN) test. The time series models were tested in four emergency department sites to assess forecasting accuracy using the root mean square error and mean absolute percentage error. The models were tested for (i) short term prediction (four weeks ahead), using weekday time series; and (ii) long term predictions (four months ahead) using monthly time series. Data analysis revealed that presentations to emergency department and subsequent admissions to hospital were not a purely random process and therefore could be predicted with acceptable accuracy. Prediction accuracy improved as the forecast time intervals became wider (from daily to monthly). For each weekday time series modelling using fuzzy time series, for forecasting daily admissions, the mean absolute percentage error ranged from 2.63% to 4.72% while for monthly time series mean absolute percentage error varied from 2.01%-2.81%. For weekday time series, the mean absolute percentage error for autoregressive integrated moving average and neural network forecasting models ranged from 6.25% to 7.47% and 6.04%-7.42% respectively. The proposed fuzzy time series model proved to have statistically significant performance using Harvey, Leybourne and Newbold (HLN) test. This was explained by variations in attendances in different sites and weekdays. This paper described a heuristic-based fuzzy logic model for predicting emergency department attendances which could help resource allocation and reduce pressure on busy hospitals. Valid and reproducible prediction tools could be generated from these hospital data. The methodology had an acceptable accuracy over a relatively short time period, and could be used to assist better bed management, staffing and elective surgery scheduling. When compared to other prediction models usually applied for emergency department attendances prediction, the proposed heuristic model had better accuracy.


Effectiveness of mailed letters to improve medication adherence among Medicare Advantage Plan participants with chronic conditions.
Medication adherence is associated with improved health outcomes in multiple chronic diseases. Information is needed on the effectiveness of specific adherence interventions. This study's objectives were to quantify effects of a targeted mailing intervention on adherence among older adults at risk for nonadherence, and to examine associations of individual and plan characteristics with adherence. Among adults enrolled in a Medicare Advantage Plan with prescription drug coverage from May 2014 to June 2015, those identified as eligible for the mailing intervention had a late refill for oral antidiabetic medication, statin, angiotensin-converting enzyme inhibitor, or angiotensin receptor blocker medication and were previously unreachable by telephone. Pharmacy claims data were analyzed with the outcome of 6-month proportion of days covered (PDC) before and after the mailing. The t-test and chi-square analyses were used to evaluate univariate associations. Multivariable linear and logistic regression models were conducted to assess relative covariate effects. A sub-analysis of those with at least one medication fill post-mailing was also performed. A total of 460 non-adherent individuals aged 70±10.5 years, with 50.2% female and 66.7% white individuals, were included. Of those who were mailed a letter, 24.1% became adherent to the specified maintenance medication. Those who received >30-day supplies were more than twice as likely to become adherent after the mailed letter than those who received 30-day supplies or less (P<0.05). Baseline higher PDC was also associated with greater adherence post-mailing (P<0.01). A total of 284 (61.7%) individuals filled their medication at least once after the mailed letter; of those, 39.1% became adherent (mean [SD] change in PDC =0.15 [±0.28]). Our findings suggest that a single mailed letter improved medication adherence by 24.1% in adults with chronic conditions. As a health plan seeking to improve its customers' well-being and outcomes, Cigna continues to utilize targeted mail interventions to improve medication adherence.


Neuroimaging Findings in Cardiac Myxoma Patients: A Single-Center Case Series of 47 Patients.
Cardiac myxomas can present with a myriad of neurological complications including stroke, cerebral aneurysm formation and metastatic disease. Our study had two objectives: (1) to describe the neuroimaging findings of patients with cardiac myxomas and (2) to examine the relationship between a history of embolic complications secondary to myxoma and intracranial aneurysm formation, hemorrhage and metastatic disease. We hypothesized that patients who present with embolic complications related to myxoma would be more likely to have such complications. We searched our institutional database for all patients with pathologically proven cardiac myxomas from 1995 to 2014 who received neuroimaging. Neuroimaging findings were categorized as acute ischemic stroke, intracerebral hemorrhage, oncotic aneurysm, and cerebral metastasis. Cardiac myxoma patients were divided into those presenting with embolic complications (i.e. lower extremity emboli or cerebral emboli) and those presenting with non-embolic complications prior to surgical resection of the myxoma. The prevalence of intracranial hemorrhage, myxomatous aneurysm formation, and cerebral metastases was compared in myxoma patients presenting with and without embolic complications using a Chi-squared test. Forty-seven consecutive patients were included in this study. Sixteen patients (34.0%) had imaging evidence of acute ischemic stroke. Of these, 13 had acute ischemic strokes directly attributed to the cardiac myxoma (27.7%) and 3 had acute ischemic strokes secondary to causes other than myxoma (6.4%). Seven patients (14.9%) had aneurysms. Two patients (4.3%) had parenchymal metastatic disease on long-term imaging. Fourteen patients (29.8%) presented with ischemic symptoms that were attributed to cardiac myxoma (1 with lower extremity ischemia, 1 with lower extremity ischemia and ischemic stroke, and 12 with ischemic stroke). Patients presenting with embolic complications related to the myxoma (ischemic stroke or lower extremity ischemia) were more likely to have imaging evidence of intracranial hemorrhage (21.4 vs. 3.0%, p = 0.09), oncotic aneurysm (35.7 vs. 6.1%, p = 0.03), and cerebral metastasis (14.3 vs. 0.0%, p = 0.07) on follow-up imaging. Ischemic stroke and intracranial oncotic aneurysm were found in a substantial proportion of cardiac myxoma patients undergoing neuroimaging. Patients presenting with embolic complications of cardiac myxoma are more likely to have intracranial hemorrhage, intracranial oncotic aneurysms, and cerebral metastatic disease.


Divergence between Drosophila santomea and allopatric or sympatric populations of D. yakuba using paralogous amylase genes and migration scenarios along the Cameroon volcanic line.
We have used two paralogous genes (Amyrel and Amy) of the amylase multigene family to reconstruct the phylogeny of the nine Drosophila melanogaster subgroup sister species, including D. santomea, the newly discovered endemic from São Tomé island. The evolutionary divergence of these genes is of special interest as it is suspected to result from physiological evolution via gene duplication. This paper describes the relationship between the geographical origin of the various strains and the patterns of mating and phylogeny, focusing on the evolution of D. santomea and its relationship to other species and their niches. The Amyrel and Amy data indicate that, contrary to expectations, the sympatric insular D. yakuba population is less closely related to D. santomea than allopatric mainland ones, suggesting that the extant insular D. yakuba population on São Tomé results from a recent secondary colonization. Data for sympatric and allopatric D. yakuba suggest that D. santomea arose from a mainland D. yakuba parental stock when montane habitats of the Cameroon volcanic line extended to lower altitudes during colder and less humid periods. Despite their different modes of evolution and different functions, the Amyrel and Amy genes provide remarkably consistent topologies and hence reflect the same history, that of the species.


Neurogenic origin of articular hyperemia in early degenerative joint disease.
It has been speculated that joint instability resulting from anterior cruciate ligament (ACL) rupture could be exacerbated by changes in vasomotor activity in the remaining supporting structures. In this study, the effect of ACL transection on medial collateral ligament (MCL) basal perfusion and its responsiveness to calcitonin gene-related peptide (CGRP) and sympathetic adrenergic influences was examined. Using urethan-anesthetized rabbits, we tested the effects of CGRP and its antagonist CGRP-(8-37) by topical application of these agents to the exposed knee while sympathetic influences were tested by electrically stimulating the saphenous nerve. It was found that MCL basal perfusion was elevated in ACL-sectioned joints; however, this effect was abrogated by prior resection of the articular nerve supply. At the doses tested, the normal vasodilator response to CGRP was abolished in ACL-sectioned joints, whereas the response to CGRP-(8-37) was attenuated. Even under the influence of increased constrictor tone, MCL and capsule blood vessels still showed substantially reduced responses to exogenous CGRP administration. By contrast, nerve-mediated constrictor responses were mostly unaffected by joint instability. This study suggests that posttraumatic knee joint hyperemia is neurogenically mediated, possibly by increased secretion of CGRP.


Spontaneous spatiotemporal ordering of shape oscillations enhances cell migration.
The migration of cells is relevant for processes such as morphogenesis, wound healing, and invasion of cancer cells. In order to move, single cells deform cyclically. However, it is not understood how these shape oscillations influence collective properties. Here we demonstrate, using numerical simulations, that the interplay of directed motion, shape oscillations, and excluded volume enables cells to locally "synchronize" their motion and thus enhance collective migration. Our model captures elongation and contraction of crawling ameboid cells controlled by an internal clock with a fixed period, mimicking the internal cycle of biological cells. We show that shape oscillations are crucial for local rearrangements that induce ordering of neighboring cells according to their internal clocks even in the absence of signaling and regularization. Our findings reveal a novel, purely physical mechanism through which the internal dynamics of cells influences their collective behavior, which is distinct from well known mechanisms like chemotaxis, cell division, and cell-cell adhesion.


Analysis of primary care electronic health record data of people living with hepatitis B virus: infection and hepatocellular carcinoma risk associated with socio-economic deprivation.
We set out to characterise chronic hepatitis B (CHB) in the primary care population in England and investigate risk factors for progression to hepatocellular carcinoma (HCC). Retrospective cohort study. We identified 8039 individuals with CHB in individuals aged ≥18 years between 1999 and 2019 in the English primary care database QResearch. HCC risk factors were investigated using Cox proportional hazards modelling. Most of those with a record of CHB were males (60%) of non-White ethnicity (>70%), and a high proportion were in the most deprived Townsend deprivation quintile (44%). Among 7029 individuals with longitudinal data, 161 HCC cases occurred. Increased HCC hazards were significantly associated with male sex (adjusted hazards ratio [aHR] 3.17, 95% confidence interval [95% CI] 1.92-5.23), in the fifth deprivation quintile as compared to the third quintile (aHR 1.69, 95% CI 1.01-2.84), with older age (for age groups 56-65 and ≥66 years, compared to 26-35 years, aHRs 2.82 [95% CI 1.45-5.46] and 3.76 [95% CI 1.79-7.9], respectively), Caribbean ethnicity (aHR 3.32, 95% CI 1.43-7.71, compared to White ethnicity), ascites (aHR 3.15, 95% CI 1.30-7.67), cirrhosis (aHR 6.55, 95% CI 4.57-9.38) and peptic ulcer disease (aHR 2.26, 95% CI 1.45-3.51). Targeting interventions and HCC surveillance at vulnerable groups is essential to improve CHB outcomes and to support progress towards international goals for the elimination of hepatitis infection as a public health threat.


The endocannabinoid system and the brain.
The psychoactive constituent in cannabis, Δ(9)-tetrahydrocannabinol (THC), was isolated in the mid-1960s, but the cannabinoid receptors, CB1 and CB2, and the major endogenous cannabinoids (anandamide and 2-arachidonoyl glycerol) were identified only 20 to 25 years later. The cannabinoid system affects both central nervous system (CNS) and peripheral processes. In this review, we have tried to summarize research--with an emphasis on recent publications--on the actions of the endocannabinoid system on anxiety, depression, neurogenesis, reward, cognition, learning, and memory. The effects are at times biphasic--lower doses causing effects opposite to those seen at high doses. Recently, numerous endocannabinoid-like compounds have been identified in the brain. Only a few have been investigated for their CNS activity, and future investigations on their action may throw light on a wide spectrum of brain functions.


Profiles of cognitive dysfunction in chronic amphetamine and heroin abusers.
Groups of subjects whose primary drug of abuse was amphetamine or heroin were compared, together with age- and IQ-matched control subjects. The study consisted of a neuropsychological test battery which included both conventional tests and also computerised tests of recognition memory, spatial working memory, planning, sequence generation, visual discrimination learning, and attentional set-shifting. Many of these tests have previously been shown to be sensitive to cortical damage (including selective lesions of the temporal or frontal lobes) and to cognitive deficits in dementia, basal ganglia disease, and neuropsychiatric disorder. Qualitative differences, as well as some commonalities, were found in the profile of cognitive impairment between the two groups. The chronic amphetamine abusers were significantly impaired in performance on the extra-dimensional shift task (a core component of the Wisconsin Card Sort Test) whereas in contrast, the heroin abusers were impaired in learning the normally easier intra-dimensional shift component. Both groups were impaired in some of tests of spatial working memory. However, the amphetamine group, unlike the heroin group, were not deficient in an index of strategic performance on this test. The heroin group failed to show significant improvement between two blocks of a sequence generation task after training and additionally exhibited more perseverative behavior on this task. The two groups were profoundly, but equivalently impaired on a test of pattern recognition memory sensitive to temporal lobe dysfunction. These results indicate that chronic drug use may lead to distinct patterns of cognitive impairment that may be associated with dysfunction of different components of cortico-striatal circuitry.


Differential diagnoses and management strategies in patients with schizophrenia and bipolar disorder.
Successful treatment of psychiatric disorders, including bipolar disorder and schizophrenia, is complicated and is affected by a broad range of factors associated with the diagnosis, choice of treatment and social factors. In these patients, treatment management must focus on accurate and early diagnosis, to ensure that correct treatment is administered as soon as possible. In both disorders, the treatment of the disease in the acute phase must be maintained long term to provide continuous relief and normal function; the treatment choice in the early stages of the disease may impact on long-term outcomes. In schizophrenia, treatment non-compliance is an important issue, with up to 50% of patients discontinuing treatment for reasons as diverse as efficacy failure, social barriers, and more commonly, adverse events. Treatment non-compliance also remains an issue in bipolar disorder, as tolerability of mood stabilizers, especially lithium, is not always good, and combination treatments are frequent. In order to achieve an optimal outcome in which the patient continues with their medication life-long, treatment should be tailored to each individual, taking into account treatment and family history, and balancing efficacy with tolerability to maximize patient benefit and minimize the risk of discontinuation. These case studies illustrate how treatment should be monitored, tailored and often changed over time to meet these needs.


Comparison of cutaneous and bronchial reactivity to leukotriene D4 in humans.
To establish whether a relationship exists between cutaneous and bronchial reactivity to leukotriene D4 (LTD4), endpoint cutaneous reactivity and bronchial reactivity were determined in six patients with asthma, six patients with seasonal allergic rhinitis, and six nonatopic men. In all subjects, increasing doses of LTD4 produced decreases in specific airway conductance (SGaw) and flow at 30% of vital capacity measured from a partial forced expiratory maneuver (V30P). Intracutaneous injections of LTD4 and histamine resulted in immediate wheal and erythema reactions in all patients. Serial 10-fold dilutions of LTD4 and histamine were injected to establish the lowest dose capable of producing a 4 mm wheal with erythema. Endpoint cutaneous reactivity to LTD4 ranged from 1 mumol to 100 pmol in controls and from 100 nmol to 1 fmol in patients with asthma, but statistically significant differences between groups were not apparent even with inclusion of six additional subjects. No meaningful correlations were found between the quantity of LTD4 producing the threshold cutaneous reaction and the dose of inhaled LTD4 that produced a 35% decrease in SGaw and 30% decrease in V30P. In contrast to the bronchi, where subjects in all three groups were 264 to 840 times more sensitive to LTD4 than to methacholine, 11/18 subjects (3/6 with asthma) were less sensitive to LTD4 than to histamine in terms of endpoint cutaneous reactivity. However, seven of 18 subjects who were more sensitive to LTD4 than to histamine had relative potencies ranging from 5 to 500,000.


Artificial intelligence in health care: accountability and safety.
The prospect of patient harm caused by the decisions made by an artificial intelligence-based clinical tool is something to which current practices of accountability and safety worldwide have not yet adjusted. We focus on two aspects of clinical artificial intelligence used for decision-making: moral accountability for harm to patients; and safety assurance to protect patients against such harm. Artificial intelligence-based tools are challenging the standard clinical practices of assigning blame and assuring safety. Human clinicians and safety engineers have weaker control over the decisions reached by artificial intelligence systems and less knowledge and understanding of precisely how the artificial intelligence systems reach their decisions. We illustrate this analysis by applying it to an example of an artificial intelligence-based system developed for use in the treatment of sepsis. The paper ends with practical suggestions for ways forward to mitigate these concerns. We argue for a need to include artificial intelligence developers and systems safety engineers in our assessments of moral accountability for patient harm. Meanwhile, none of the actors in the model robustly fulfil the traditional conditions of moral accountability for the decisions of an artificial intelligence system. We should therefore update our conceptions of moral accountability in this context. We also need to move from a static to a dynamic model of assurance, accepting that considerations of safety are not fully resolvable during the design of the artificial intelligence system before the system has been deployed.


Proteomic serum profile in menstrual-related and post menopause migraine.
The aim of this pilot study was to analyze the serum proteomic profile of women suffering from menstrual-related migraine (MM group, n = 15) and migraine in post-menopause (PM group, n = 15) in comparison with non-headache control females (C group, n = 15). Serum samples were subjected to two-dimensional gel electrophoresis (2-DE) followed by mass spectrometry (MS) analysis for protein identification. Based on 2D-gel maps and PDQuest 2-D software, 13 differentially expressed spots, corresponding to 12 unique proteins identified by Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight/tandem mass spectrometry (LC-ESI-QToF-MS/MS), were detected in the MM and PM groups vs C group. Five inflammatory and regulatory of vascular integrity proteins (prothrombin, serum amyloid P-component, Ig kappa chain C region, apolipoprotein A-I, serum amyloid A-4 protein) were found deregulated in both MM and PM groups compared to C group; MM group showed the upregulation of other inflammatory protein fragments (inter-alpha-trypsin inhibitor heavy chain H4 and complement C4-A) compared to C group; PM group, in comparison with C group, displayed a noteworthy upregulation of transthyretin and other deregulated proteins (tetranectin, alpha-1-antitrypsin, haptoglobin, apolipoprotein A-IV) playing a role in anti-inflammatory and reparative processes. In conclusion, proteomic analysis was able to reveal differences in protein expression between migraine sufferers and non-headache women; as in other neurological diseases characterized by neuroinflammation, the serum proteome of migraine women presents an abundance of proteins indicative of cellular damage, oxidative stress and inflammation. This relevant inflammatory status, if confirmed in larger series, could represent a target for menstrual-related migraine treatment.


Linarin Protects Against CCl4-Induced Acute Liver Injury via Activating Autophagy and Inhibiting the Inflammatory Response: Involving the TLR4/MAPK/Nrf2 Pathway.
Linarin has been implicated in the inhibition of inflammatory responses and hepatoprotective effects. However, the precise mechanism by which Linarin integrates injury-induced signaling from inflammatory responses and oxidative stress remains unclear. We evaluated the role of Linarin in a mouse model of carbon tetrachloride (CCl4)-induced acute liver injury. Mice were orally pretreated with Linarin or vehicle for seven consecutive days, followed by intraperitoneal injection with 0.2% (v/v) CCl4. To investigate the mechanism of action on oxidative stress, CCl4-stimulated HepG2 cells were utilized. Our results revealed Linarin remarkably attenuated the loss of hepatic architecture, inflammatory cell infiltration, serum transaminases, and pro-inflammatory cytokines induced by CCl4. Linarin attenuated CCl4-induced oxidative stress by increasing the expression of cytosolic Nrf2 (nuclear factor erythroid 2-related factor 2), inducing nuclear localization of Nrf2, and increasing stress-induced protein heme oxygenase-1 (HO-1). Additionally, Linarin decreased the expression of toll-like receptors (TLR)-4, and its downstream proteins, MyD88, IRAK1, and TRAF6. Furthermore, Linarin reversed CCl4-induced phosphorylation of ERK, p38, and JNK. Importantly, Linarin increased the expression of both LC3II and Beclin 1, which are hallmarks of autophagic flux. Autophagy-mediated hepatoprotective effects in Linarin-treated HepG2 cells were mitigated by the autophagy inhibitor 3-MA. However, combined treatment of Linarin with 3-MA failed to significantly reverse cell apoptosis and the production of transaminases and pro-inflammatory cytokines. Linarin prevents acute liver injury, possibly by alleviating ROS-induced oxidative stress, inhibiting TLR4/MyD88 and JNK/p38/ERK-mediated inflammatory responses, and promoting Beclin 1/LC3II-mediated autophagic flux.


Genetically distinct Group B Streptococcus strains induce varying macrophage cytokine responses.
Group B Streptococcus (GBS) is an opportunistic pathogen that causes preterm birth and neonatal disease. Although GBS is known to exhibit vast diversity in virulence across strains, the mechanisms of GBS-associated pathogenesis are incompletely understood. We hypothesized that GBS strains of different genotypes would vary in their ability to elicit host inflammatory responses, and that strains associated with neonatal disease would induce different cytokine profiles than those associated with colonization. Using a multiplexed, antibody-based protein detection array, we found that production of a discrete number of inflammatory mediators by THP-1 macrophage-like cells was universally induced in response to challenge with each of five genetically distinct GBS isolates, while other responses appeared to be strain-specific. Key array responses were validated by ELISA using the initial five strains as well as ten additional strains with distinct genotypic and phenotypic characteristics. Interestingly, IL-6 was significantly elevated following infection with neonatal infection-associated sequence type (ST)-17 strains and among strains possessing capsule (cps) type III. Significant differences in production of IL1-β, IL-10 and MCP-2 were also identified across STs and cps types. These data support our hypothesis and suggest that unique host innate immune responses reflect strain-specific differences in virulence across GBS isolates. Such data might inform the development of improved diagnostic or prognostic strategies against invasive GBS infections.


Computational and Mathematical Methods in Medicine Prediction of COVID-19 in BRICS Countries: An Integrated Deep Learning Model of CEEMDAN-R-ILSTM-Elman.
Since the outbreak of COVID-19, BRICS countries have experienced different epidemic spread due to different health conditions, social isolation measures, vaccination rates, and other factors. A descriptive analysis is conducted for the spread of the epidemic in the BRICS countries. Considering the nonlinear and nonstationary characteristics of COVID-19 data, a principle of decomposition-reconstruction(R)-prediction-integration is proposed. Correspondingly, this paper constructs an integrated deep learning prediction model of CEEMDAN-R-ILSTM-Elman. Specifically, the prediction model is integrated by complete ensemble empirical mode decomposition (CEEMDAN), improved long-term and short-term memory network (ILSTM), and Elman neural network. First, the data is decomposed by adopting CEEMDAN. Then, by calculating the permutation entropy and average period, the decomposed eigenmode component IMFs are reconstructed into four sequences of high, medium, low level, and trend term. Thus, ILSTM and Elman algorithms are used for component sequence prediction, whose results are integrated as the final results. The ILSTM is established based on the LSTM model and the improved beetle antennae search algorithm (IBAS). The ILSTM mainly considers that the prediction accuracy of LSTM model is vulnerable to the influence of parameter selection. The IBAS with adaptive step size is used to automatically optimize the super parameters of LSTM model and to improve the modeling efficiency and prediction accuracy. Experimental results indicate that compared with other benchmark models, CEEMDAN-R-ILSTM-Elman integrated model predicts the number of newly confirmed cases of COVID-19 in BRICS countries with higher accuracy and lower error. Strict social policies have a greater impact on the infection rate and mortality rate of the epidemic. During July-August 2021, epidemic spread in BRICS countries will slow down, and the overall situation is still quite severe.


Peripheral blood mononuclear cell gene expression profiles predict poor outcome in idiopathic pulmonary fibrosis.
We aimed to identify peripheral blood mononuclear cell (PBMC) gene expression profiles predictive of poor outcomes in idiopathic pulmonary fibrosis (IPF) by performing microarray experiments of PBMCs in discovery and replication cohorts of IPF patients. Microarray analyses identified 52 genes associated with transplant-free survival (TFS) in the discovery cohort. Clustering the microarray samples of the replication cohort using the 52-gene outcome-predictive signature distinguished two patient groups with significant differences in TFS. We studied the pathways associated with TFS in each independent microarray cohort and identified decreased expression of "The costimulatory signal during T cell activation" Biocarta pathway and, in particular, the genes CD28, ICOS, LCK, and ITK, results confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). A proportional hazards model, including the qRT-PCR expression of CD28, ICOS, LCK, and ITK along with patient's age, gender, and percent predicted forced vital capacity (FVC%), demonstrated an area under the receiver operating characteristic curve of 78.5% at 2.4 months for death and lung transplant prediction in the replication cohort. To evaluate the potential cellular source of CD28, ICOS, LCK, and ITK expression, we analyzed and found significant correlation of these genes with the PBMC percentage of CD4(+)CD28(+) T cells in the replication cohort. Our results suggest that CD28, ICOS, LCK, and ITK are potential outcome biomarkers in IPF and should be further evaluated for patient prioritization for lung transplantation and stratification in drug studies.


Effects of dihydropyridines on aldosterone secretion in adrenal capsule preparations from pregnant rats.
The primary aim of this study was to determine when sensitivity in the aldosterone response to extracellular potassium (K+) decreases during pregnancy. Second, it tested the hypothesis that calcium channel alterations occur in the adrenal cortex during pregnancy. The decreased sensitivity to K+, observed at 22 days of gestation, was not evident at 15 days and between 18 and 36 h postpartum. Increases in extracellular calcium concentration heightened sensitivity to K+ in adrenal capsule preparations derived from nonpregnant rats but had no effect in pregnant animals. The influence of nifedipine and BAY K 8644 (blocker and activator, respectively, of voltage-operated calcium channels) on the aldosterone response to K+ and to adrenocorticotropic hormone (ACTH) was studied. Sensitivity to K+ in nonpregnant rats decreased in the presence of nifedipine and became similar to that in pregnant rats. Responses to ACTH were not affected by nifedipine. BAY K 8644 produced a larger increase in sensitivity in adrenal capsule preparations from pregnant than from nonpregnant rats, leading to superposition of the two dose-response curves to K+. These results indicate that voltage-operated calcium channels involved in aldosterone secretion are functionally impaired during pregnancy.


The Psychometric Structure of Items Assessing Autogynephilia.
Autogynephilia, or paraphilic sexual arousal in a man to the thought or image of himself as a woman, manifests in a variety of different behaviors and fantasies. We examined the psychometric structure of 22 items assessing five known types of autogynephilia by subjecting them to exploratory factor analysis in a sample of 149 autogynephilic men. Results of oblique factor analyses supported the ability to distinguish five group factors with suitable items. Results of hierarchical factor analyses suggest that the five group factors were strongly underlain by a general factor of autogynephilia. Because the general factor accounted for a much greater amount of the total variance of the 22 items than did the group factors, the types of autogynephilia that a man has seem less important than the degree to which he has autogynephilia. However, the five types of autogynephilia remain conceptually useful because meaningful distinctions were found among them, including differential rates of endorsement and differential ability to predict other relevant variables like gender dysphoria. Factor-derived scales and subscales demonstrated good internal consistency reliabilities, and validity, with large differences found between autogynephilic men and heterosexual male controls. Future research should attempt to replicate our findings, which were mostly exploratory.


The disturbed redox-balance in pulmonary fibrosis is modulated by the plant flavonoid quercetin.
Idiopathic pulmonary fibrosis (IPF) is characterized by a disturbed pulmonary redox balance associated with inflammation. To restore this balance, antioxidants are often suggested as therapy for IPF but previous clinical trials with these compounds and their precursors have not been successful in the clinic. The exogenous antioxidant quercetin, which has a versatile antioxidant profile and is effective in restoring a disturbed redox balance, might be a better candidate. The aim of this study was to evaluate the protective effect of quercetin on oxidative and inflammatory markers in IPF. Here, we demonstrate that IPF patients have a significantly reduced endogenous antioxidant defense, shown by a reduced total antioxidant capacity and lowered glutathione and uric acid levels compared to healthy controls. This confirms that the redox balance is disturbed in IPF. Ex vivo incubation with quercetin in blood of both IPF patients and healthy controls reduces LPS-induced production of the pro-inflammatory cytokines IL-8 and TNFα. This anti-inflammatory effect was more pronounced in the blood of the patients. Our pro-fibrotic in vitro model, consisting of bleomycin-triggered BEAS-2B cells, shows that quercetin boosts the antioxidant response, by increasing Nrf2 activity, and decreases pro-inflammatory cytokine production in a concentration-dependent manner. Collectively, our findings implicate that IPF patients may benefit from the use of quercetin to restore the disturbed redox balance and reduce inflammation.


Plasmonic titanium nitride nanoparticles for in vivo photoacoustic tomography imaging and photothermal cancer therapy.
Titanium nitride, an alternative plasmonic material to gold with unique physiochemical properties, has been widely used in microelectronics, biomedical devices and food-contact applications. However, its potential application in the area of biomedicine has not been effectively explored. With the spectral match of their plasmon resonance band and the biological transparency window as well as good biocompatibility, titanium nitride nanoparticles (TiN NPs) are promising photoabsorbing agents for photothermal therapy (PTT) and photoacoustic imaging. Nevertheless, the photothermal performance of TiN NPs has not been investigated until now. Here, we presented the investigation of employing TiN NPs as photoabsorbing agents for in vivo photoacoustic tomography (PAT) imaging-guided photothermal cancer therapy. Our experimental results showed that TiN NPs could strongly absorb the NIR light and provided up to 48% photothermal conversion efficiency. After PEGylation, the resultant nanoparticles demonstrated improved physiological stability and extensive blood retention. Following intravenously administration, they could simultaneously enhance the photoacoustic signals of the tumor region and destroy tumors in the tumor-bearing mouse model by taking advantage of the photothermal effect of the TiN NPs. Our findings highlighted the great potential of plasmonic TiN NPs in detection and treatment of cancer.


Copper status of ewes fed increasing amounts of copper from copper sulfate or copper proteinate.
The Cu status of mature, crossbred ewes fed two sources (CuSO4 vs. Cu proteinate) and three levels (10, 20, or 30 mg/kg) of dietary Cu was determined in a 73-d feeding trial. Ewes (n = 30) were fed a basal diet containing rice meal feed, cottonseed hulls, cottonseed meal, meat and bone meal, cracked corn, and vitamin-mineral supplements at 2.5% of BW to meet NRC requirements for protein, energy, macrominerals, and microminerals, excluding Cu. The basal diet contained 5 mg/kg Cu, 113 mg/kg Fe, .1 mg/kg Mo, and .17% S. Copper sulfate or Cu proteinate was added to the basal diet to supply 10, 20, or 30 mg/kg of dietary copper in a 2x3 factorial arrangement of treatments. Ewes were housed in 3.7- x 9.1-m pens in an open-sided barn. Blood samples were collected on d 28 and 73. Ewes were slaughtered on d 74, and liver and other tissues were collected to determine Cu concentrations. An interaction (P = .08) occurred between source and level for liver Cu. The interaction existed due to an increase in liver Cu concentrations when ewes were fed increasing dietary Cu from CuSO4 but not when fed Cu proteinate diets. There was no source x level interaction (P>.10) for the blood constituents measured. On d 73, plasma ceruloplasmin activity was greater (P<.05) in ewes fed Cu proteinate than in those fed CuSO4 (33.1 vs. 26.8 microM x min(-1) x L(-1)). Increasing the concentration of dietary Cu did not affect (P>.10) plasma ceruloplasmin. Packed cell volume (PCV), red blood cell count (RBC), white blood cell count, whole blood hemoglobin (wHb), plasma hemoglobin, and plasma Cu were similar between sources of Cu. Ewes fed 20 mg/kg Cu had lower (P<.05) PCV, RBC, and wHb than those fed 10 or 30 mg/kg Cu diets. Feeding up to 30 mg/kg Cu from these sources did not cause an observable Cu toxicity during the 73-d period.


Effect of methionine oxidation of a recombinant monoclonal antibody on the binding affinity to protein A and protein G.
Oxidation of methionine (Met) residues is one of the most common protein degradation pathways. Two Met residues, Met256 and Met432, of a recombinant fully human monoclonal IgG1 antibody have been shown to be susceptible to oxidation. Met256 and Met432 are located in the antibody CH2-CH3 interface and in close proximity to protein A and protein G binding sites. The effect of oxidation of these susceptible Met residues on the binding to protein A and protein G was investigated in the current study. Incubation of the antibody with 5% tert-butyl hydroperoxide (tBHP) resulted in a nearly complete oxidation of Met256 and Met432, while incubation with 1% tBHP resulted in mixed populations of the antibody with different degrees of Met oxidation. Oxidation of Met256 and Met432 resulted in earlier elution of the antibody from protein A and protein G columns when eluted with a gradient of decreasing pH. Analysis by ELISA and surface plasmon resonance (SPR) revealed decreased binding affinity of the oxidized antibody to protein A and protein G. It is therefore concluded that oxidation of the Met256 and Met432 residues of the recombinant monoclonal antibody altered its interaction with protein A and protein G resulting in a decrease in binding affinity.


Modularity Facilitates Classification Performance of Spiking Neural Networks for Decoding Cortical Spike Trains.
After the introduction of recurrence, an important property of the biological brain, spiking neural networks (SNNs) have achieved unprecedented classification performance. But they still cannot outperform many artificial neural networks. Modularity is another crucial feature of the biological brain. It remains unclear if modularity can also improve the performance of SNNs. To investigate this idea, we proposed the modular SNN, and compared its performance with a uniform SNN without modularity by employing them to classify cortical spike trains. For the first time, a significant improvement was found in our modular SNN. Further, we probed into the factors influencing the performance of the modular SNN and found: (a). The modular SNN outperformed the uniform SNN more significantly when the number of neurons in the networks increased; (b). The performance of the modular SNNs increased as the number of modules dropped. These preliminary but novel findings suggest that modularity may help develop better artificial intelligence and brain-machine interfaces. Also, the modular SNN may serve as a model for the study of neuronal spike synchrony.


EMDR effects on pursuit eye movements.
This study aimed to objectivize the quality of smooth pursuit eye movements in a standard laboratory task before and after an Eye Movement Desensitization and Reprocessing (EMDR) session run on seven healthy volunteers. EMDR was applied on autobiographic worries causing moderate distress. The EMDR session was complete in 5 out of the 7 cases; distress measured by SUDS (Subjective Units of Discomfort Scale) decreased to a near zero value. Smooth pursuit eye movements were recorded by an Eyelink II video system before and after EMDR. For the five complete sessions, pursuit eye movement improved after their EMDR session. Notably, the number of saccade intrusions-catch-up saccades (CUS)-decreased and, reciprocally, there was an increase in the smooth components of the pursuit. Such an increase in the smoothness of the pursuit presumably reflects an improvement in the use of visual attention needed to follow the target accurately. Perhaps EMDR reduces distress thereby activating a cholinergic effect known to improve ocular pursuit.


Identifying target organ location of Radix Achyranthis Bidentatae: a bioinformatics approach on active compounds and genes.
Background: Herbal medicines traditionally target organs for treatment based on medicinal properties, and this theory is widely used for prescriptions. However, the scientific evidence explaining how herbs act on specific organs by biological methods has been still limited. This study used bioinformatic tools to identify the target organ locations of Radix Achyranthis Bidentatae (RAB), a blood-activating herb that nourishes the liver and kidney, strengthens bones, and directs prescription to the lower body. Methods: RAB's active compounds and targets were collected and predicted using databases such as TCMSP, HIT2.0, and BATMAN-TCM. Next, the RAB's target list was analyzed based on two approaches to obtain target organ locations. DAVID and Gene ORGANizer enrichment-based approaches were used to enrich an entire gene list, and the BioGPS and HPA gene expression-based approaches were used to analyze the expression of core genes. Results: RAB's targets were found to be involved in whole blood, blood components, and lymphatic organs across all four tools. Each tool indicated a particular aspect of RAB's target organ locations: DAVID-enriched genes showed a predominance in blood, liver, and kidneys; Gene ORGANizer showed the effect on low body parts as well as bones and joints; BioGPS and HPA showed high gene expression in bone marrow, lymphoid tissue, and smooth muscle. Conclusion: Our bioinformatics-based target organ location prediction can serve as a modern interpretation tool for the target organ location theory of traditional medicine. Future studies should predict therapeutic target organ locations in complex prescriptions rather than single herbs and conduct experiments to verify predictions.


Genetic interactions between Cdk1-CyclinB and the Separase complex in Drosophila.
Cdk1-CycB plays a key role in regulating many aspects of cell-cycle events, such as cytoskeletal dynamics and chromosome behavior during mitosis. To investigate how Cdk1-CycB controls the coordination of these events, we performed a dosage-sensitive genetic screen, which is based on the observations that increased maternal CycB (four extra gene copies) leads to higher Cdk1-CycB activity in early Drosophila embryos, delays anaphase onset, and generates a sensitized non-lethal phenotype at the blastoderm stage (defined as six cycB phenotype). Here, we report that mutations in the gene three rows (thr) enhance, while mutations in pimples (pim, encoding Drosophila Securin) or separase (Sse) suppress, the sensitized phenotype. In Drosophila, both Pim and Thr are known to regulate Sse activity, and activated Sse cleaves a Cohesin subunit to initiate anaphase. Compared with the six cycB embryos, reducing Thr in embryos with more CycB further delays the initiation of anaphase, whereas reducing either Pim or Sse has the opposite effect. Furthermore, nuclei move slower during cortical migration in embryos with higher Cdk1-CycB activity, whereas reducing either Pim or Sse suppresses this phenotype by causing a novel nuclear migration pattern. Therefore, our genetic screen has identified all three components of the complex that regulates sister chromatid separation, and our observations indicate that interactions between Cdk1-CycB and the Pim-Thr-Sse complex are dosage sensitive.


Dual therapy with peg-interferon and ribavirin in thalassemia major patients with chronic HCV infection: Is there still an indication?
Iron overload and hepatitis C virus (HCV) infection together can lead to chronic liver damage in thalassemia major (TM) patients. We investigated viral, genetic, and disease factors influencing sustained virological response (SVR) after peg-interferon and ribavirin therapy in TM patients with HCV infection. We analyzed 230 TM patients with HCV infection (mean age 36.0±6.3 years; 59.1% genotype 1; 32.2% genotype 2; 3.4% genotype 3; and 5.3% genotype 4; 28.7% carried CC allele of rs12979860 in IL28B locus; 79.6% had chronic hepatitis and 20.4% cirrhosis; 63.5% naive and 36.5% previously treated with interferon alone) treated in 14 Italian centers. By multivariate regression analysis SVR was independently associated with CC allele of IL28B SNP (OR 2.98; CI 95% 1.29-6.86; p=0.010) and rapid virologic response (OR 11.82; CI 95% 3.83-36.54; p<0.001) in 136 genotype 1 patients. Combining favorable variables the probability of SVR ranged from 31% to 93%. In genotype 2 patients, only RVR (OR 8.61; CI 95% 2.85-26.01; p<0.001) was associated with SVR higher than 80%. In 3 patients with cirrhosis a decompensation of liver or heart disease were observed. Over 50% of patients increased blood transfusions. Dual therapy in TM patients with chronic HCV infection is efficacious in patients with the best virological, genetic and clinical predictors. Patients with cirrhosis have an increased risk of worsening liver or heart disease.


Urban bat communities are affected by wetland size, quality, and pollution levels.
Wetlands support unique biota and provide important ecosystem services. These services are highly threatened due to the rate of loss and relative rarity of wetlands in most landscapes, an issue that is exacerbated in highly modified urban environments. Despite this, critical ecological knowledge is currently lacking for many wetland-dependent taxa, such as insectivorous bats, which can persist in urban areas if their habitats are managed appropriately. Here, we use a novel paired landscape approach to investigate the role of wetlands in urban bat conservation and examine local and landscape factors driving bat species richness and activity. We acoustically monitored bat activity at 58 urban wetlands and 35 nonwetland sites (ecologically similar sites without free-standing water) in the greater Melbourne area, southeastern Australia. We analyzed bat species richness and activity patterns using generalized linear mixed-effects models. We found that the presence of water in urban Melbourne was an important driver of bat species richness and activity at a landscape scale. Increasing distance to bushland and increasing levels of heavy metal pollution within the waterbody also negatively influenced bat richness and individual species activity. Areas with high levels of artificial night light had reduced bat species richness, and reduced activity for all species except those adapted to urban areas, such as the White-striped free-tailed bat (Austronomus australis). Increased surrounding tree cover and wetland size had a positive effect on bat species richness. Our findings indicate that wetlands form critical habitats for insectivorous bats in urban environments. Large, unlit, and unpolluted wetlands flanked by high tree cover in close proximity to bushland contribute most to the richness of the bat community. Our findings clarify the role of wetlands for insectivorous bats in urban areas and will also allow for the preservation, construction, and management of wetlands that maximize conservation outcomes for urban bats and possibly other wetland-dependent and nocturnal fauna.


Smartamine M and MetaSmart supplementation during the peripartal period alter hepatic expression of gene networks in 1-carbon metabolism, inflammation, oxidative stress, and the growth hormone-insulin-like growth factor 1 axis pathways.
Peripartal cows likely require greater amounts of Met not only at the tissue and cell level for methylation reactions but also for milk protein synthesis after calving. Thirty-nine Holstein cows were fed throughout the peripartal period (-21 d to 30 d in milk) a basal control (CON) diet (n=14) with no Met supplementation, CON plus MetaSmart (MS; Adisseo Inc., Antony, France; n=12), or CON plus Smartamine M (SM; Adisseo Inc.; n=13). The Met supplements were adjusted daily and top-dressed over the total mixed ration at a rate of 0.19 or 0.07% (dry matter) of feed for MS or SM. Liver tissue was collected on -10, 7, and 21 d for transcriptome profiling of genes associated with Met and glutathione metabolism as well as components of the inflammation, oxidative stress, growth hormone/insulin-like growth factor-1 axis, and DNA methylation pathways. Data were analyzed using PROC MIXED of SAS (SAS Institute Inc., Cary, NC) with the preplanned contrasts CON versus SM + MS and SM versus MS. The S-adenosylhomocysteine hydrolase (SAHH) gene was the most abundant among all genes evaluated, with overall greater expression in Met-supplemented cows than CON, and in SM than MS. Expression of Met adenosyltransferase 1A (MAT1A) was greater in Met-supplemented cows than CON by 21 d postpartum. A greater overall expression of 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR) occurred in Met-supplemented cows than CON. In contrast, the expression of glutathione synthase (GSS); glutamate-cysteine ligase, catalytic subunit (GCLC); and superoxide dismutase 1, cytosolic (SOD1) was lower in Met-supplemented cows than CON. A greater overall expression of nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NFKB1) and greater upregulation of haptoglobin (HP) on d 7 occurred in Met-supplemented cows than CON. Expression of DNA cytosine-5-methyltransferase 3 alpha (DNMT3A) was greater but expression of DNMT1 was lower in Met-supplemented cows than CON. The response observed in SAHH reflects its importance to Met supplementation during the peripartum period. Despite greater HP expression after calving, the lower expression of glutathione (GSS and GCLC) metabolism genes and SOD1 due to Met reflect a lower oxidative stress and mild inflammatory status. The extent to which changes in expression of DNMT3A and DNMT1 result in epigenetic effects partly responsible for the previously observed enhanced performance in Met-supplemented cows remains to be examined. Increasing the supply of Met as SM or MS can affect expression of genes in the Met cycle to various extents and, hence, the supply of methyl donors such as S-adenosylmethionine and antioxidants such as glutathione. These compounds likely are in high demand during the peripartum period.


Effect of functionalization on drug delivery potential of carbon nanotubes.
The main aim of the present investigation was to explore the effect of functionalization on drug delivery potential of carbon nanotubes (CNTs) and to compare the in vitro and in vivo cancer targeting potential of doxorubicin HCL (DOX)-loaded ox-/multi-walled CNTs (MWCNTs), DOX-loaded PEG-MWCNTs and DOX-loaded FA-PEG-MWCNTs. The DOX/PEG-FA-MWCNTs showed enhanced cytotoxicity and were most preferentially taken up by the cancerous cells. The obtained results also support the extended resistance time and sustained release profile of drug-loaded surface-engineered MWCNTs. Overall, we concluded that the developed MWCNTs nanoformulations have higher cancer targeting potential.


Comparative investigations into the growth details of Histoplasma capsulatum var farciminosum on four different agar media.
Epizootic equine lymphangitis (EEL) is a chronic fungal disease that affects equids. The causative agent is a dimorphic fungus called Histoplasma capsulatum var farciminosum. H. capsulatum var farciminosum field strain 7 (D 2878/2023) isolated from the eye socket of an EEL Ethiopian horse was sub-cultured on 4 different solid media and incubated at 26°C and 37°C for 6 weeks. Details of growth morphology were recorded and shown in images during 6 weeks of incubation. H. capsulatum var farciminosum grew best at 26°C on all 4 agars, but only on sheep blood agar at 37°C as small, white dry colonies.


Epoxyeicosatrienoic acid agonist regulates human mesenchymal stem cell-derived adipocytes through activation of HO-1-pAKT signaling and a decrease in PPARγ.
Human mesenchymal stem cells (MSCs) expressed substantial levels of CYP2J2, a major CYP450 involved in epoxyeicosatrienoic acid (EET) formation. MSCs synthesized significant levels of EETs (65.8 ± 5.8 pg/mg protein) and dihydroxyeicosatrienoic acids (DHETs) (15.83 ± 1.62 pg/mg protein), suggesting the presence of soluble epoxide hydrolase (sEH). The addition of an sEH inhibitor to MSC culture decreased adipogenesis. EETs decreased MSC-derived adipocytes in a concentration-dependent manner, 8,9- and 14,15-EET having the maximum reductive effect on adipogenesis. We examined the effect of 12-(3-hexylureido)dodec-8(Z)-enoic acid, an EET agonist, on MSC-derived adipocytes and demonstrated an increased number of healthy small adipocytes, attenuated fatty acid synthase (FAS) levels (P < 0.01), and reduced PPARγ, C/EBPα, FAS, and lipid accumulation (P < 0.05). These effects were accompanied by increased levels of heme oxygenase (HO)-1 and adiponectin (P < 0.05), and increased glucose uptake (P < 0.05). Inhibition of HO activity or AKT by tin mesoporphyrin (SnMP) and LY2940002, respectively, reversed EET-induced inhibition of adipogenesis, suggesting that activation of the HO-1-adiponectin axis underlies EET effect in MSCs. These findings indicate that EETs decrease MSC-derived adipocyte stem cell differentiation by upregulation of HO-1-adiponectin-AKT signaling and play essential roles in the regulation of adipocyte differentiation by inhibiting PPARγ, C/EBPα, and FAS and in stem cell development. These novel observations highlight the seminal role of arachidonic acid metabolism in MSCs and suggest that an EET agonist may have potential therapeutic use in the treatment of dyslipidemia, diabetes, and the metabolic syndrome.


The smoothness index: a new, reproducible and clinically relevant measure of the homogeneity of the blood pressure reduction with treatment for hypertension.
To introduce a new method, the smoothness index, for assessing the homogeneity of 24 h blood pressure reduction by antihypertensive treatment and to compare it with the trough : peak ratio; and to assess the ability of both indices to predict a reduction in the left ventricular mass index induced by treatment. In 174 patients with essential hypertension and left ventricular hypertrophy, enrolled in the Study on Ambulatory Monitoring of Pressure and Lisinopril Evaluation (SAMPLE), aged 20-65 years, we measured clinic blood pressure, 24 h ambulatory blood pressure and the left ventricular mass index (echocardiography) before and after treatment with lisinopril at 20 mg with the addition of 12.5 or 25 mg hydrochlorothiazide as needed to reach a sufficient blood pressure reduction. The following parameters were computed for systolic and diastolic ambulatory blood pressure: (1) hourly and 24 h blood pressure averages (+/- SD) at baseline and after 3 and 12 months of treatment; (2) hourly blood pressure changes from baseline after 3 and 12 months of treatment, and their average (+/- SD) over 24 h; (3) the trough : peak ratio after 3 and 12 months of treatment; and (4) the smoothness index after 3 and 12 months of treatment Similar calculations were also performed at the end of a final study month during which active treatment was withdrawn and placebo was substituted (n = 164). The smoothness index for systolic and diastolic ambulatory blood pressure computed after 3 months of treatment was more closely related to its corresponding values after 12 months of treatment than the trough : peak ratio values computed after the same time periods were (r = 0.68 versus 0.38 for systolic and 0.68 versus 0.42 for diastolic blood pressure, respectively). The smoothness index showed an inverse correlation with the 24 h standard deviation of systolic and diastolic blood pressure (r = -0.25 and -0.16, P < 0.01 and < 0.05, respectively, for 12 months of treatment), while the trough : peak ratio did not (r = -0.01 to -0.12, NS). A treatment-induced reduction in the left ventricular mass index was related to the smoothness index for systolic and diastolic blood pressure (r = -0.35 and -0.32, P< 0.001 with 12 months of treatment), but not to the corresponding trough : peak ratios. The smoothness index identifies the occurrence of a balanced 24 h blood pressure reduction with treatment and correlates with the favourable effects of treatment on left ventricular hypertrophy better than the commonly used trough : peak ratio.