Diurnal rhythmicity in intestinal SGLT-1 function, V(max), and mRNA expression topography.
Mechanisms underlying the circadian rhythmicity in intestinal sugar absorption remain unclear. To test whether this rhythmicity is caused by changes in Na(+)-glucose cotransporter 1 (SGLT-1) function, we measured phloridzin-inhibitable sugar fluxes as an index of SGLT-1 activity. Jejunum obtained from rats killed at 6-h intervals during a 12-h light-dark cycle (CT0 is circadian time 0 h, time of light onset) were mounted in Ussing chambers, and 3-O-methylglucose (3-OMG) fluxes were calculated before and after addition of phloridzin. 3-OMG-induced change in short-circuit current and absorptive flux were significantly greater at CT9 than at CT3. This increase was phloridzin inhibitable. Kinetic studies indicated a significant increase in SGLT-1 maximal velocity (V(max)) at CT9. Food intake between CT3 and CT9 was <10% of the daily total, indicating that the increased SGLT-1 activity was anticipatory. Diurnicity of SGLT-1 mRNA was confirmed by Northern blotting. Expression topography analyzed by in situ hybridization revealed more intense labeling along the entire villus axis at CT9 and CT15 compared with CT3 and CT21. We conclude that diurnicity in intestinal sugar absorption is caused by periodicity in SGLT-1 V(max).


Alpha-lipoic acid treatment ameliorates metabolic parameters, blood pressure, vascular reactivity and morphology of vessels already damaged by streptozotocin-diabetes.
The present study investigated the effects of alpha-lipoic acid treatment (50 mg/kg/day) on the metabolism and vascular condition already damaged by streptozotocin (STZ)-diabetes in rats. Carbohydrate and lipid metabolism, oxidative stress and antioxidant status were assessed in non-diabetic controls, 12-week untreated diabetic and 12-week treated diabetic (untreated for 6 weeks and then treated with alpha-lipoic acid for the last 6 weeks) rats. Blood pressures of rats were measured by tail-cuff method. Vascular reactivity was evaluated in isolated aortic rings. Morphology of aorta was examined by electron microscopy technique. Alpha-lipoic acid treatment effectively reversed body weight, blood glucose, plasma insulin, cholesterol, triglycerides and lipid peroxidation levels of diabetic animals. STZ-diabetes resulted in increased blood pressure, which was partially improved by alpha-lipoic acid treatment. Although the superoxide dismutase (SOD) activity in aortic homogenates was not changed by diabetes or antioxidant treatment, catalase or glutathione peroxidase (GPx) activity significantly increased in untreated diabetic rats. Alpha-lipoic acid treatment improved catalase activity in diabetic aorta. The contractile effect of phenylephrine markedly increased in diabetic rings, which was completely reversed by alpha-lipoic acid treatment. The maximum vasorelaxant response of pre-contracted aortic rings exposed to cumulatively increased concentrations of acetylcholine was unaffected by diabetes or antioxidant treatment. Sodium nitroprusside-induced endothelium-independent relaxations were similar in all experimental groups. Various alterations caused by STZ-diabetes in aorta structure were partially ameliorated by alpha-lipoic acid treatment. The potency of alpha-lipoic acid on the reversal of hypertension by affecting vascular reactivity and morphology as well as general metabolism of diabetic rats confirms the importance of hyperglycemia-induced oxidative stress in the development of diabetes-induced vascular complications and suggests a potential therapeutic approach.


Integrative Comparison of the Role of the PHOSPHATE RESPONSE1 Subfamily in Phosphate Signaling and Homeostasis in Rice.
Phosphorus (P), an essential macronutrient for all living cells, is indispensable for agricultural production. Although Arabidopsis (Arabidopsis thaliana) PHOSPHATE RESPONSE1 (PHR1) and its orthologs in other species have been shown to function in transcriptional regulation of phosphate (Pi) signaling and Pi homeostasis, an integrative comparison of PHR1-related proteins in rice (Oryza sativa) has not previously been reported. Here, we identified functional redundancy among three PHR1 orthologs in rice (OsPHR1, OsPHR2, and OsPHR3) using phylogenetic and mutation analysis. OsPHR3 in conjunction with OsPHR1 and OsPHR2 function in transcriptional activation of most Pi starvation-induced genes. Loss-of-function mutations in any one of these transcription factors (TFs) impaired root hair growth (primarily root hair elongation). However, these three TFs showed differences in DNA binding affinities and messenger RNA expression patterns in different tissues and growth stages, and transcriptomic analysis revealed differential effects on Pi starvation-induced gene expression of single mutants of the three TFs, indicating some degree of functional diversification. Overexpression of genes encoding any of these TFs resulted in partial constitutive activation of Pi starvation response and led to Pi accumulation in the shoot. Furthermore, unlike OsPHR2-overexpressing lines, which exhibited growth retardation under normal or Pi-deficient conditions, OsPHR3-overexpressing plants exhibited significant tolerance to low-Pi stress but normal growth rates under normal Pi conditions, suggesting that OsPHR3 would be useful for molecular breeding to improve Pi uptake/use efficiency under Pi-deficient conditions. We propose that OsPHR1, OsPHR2, and OsPHR3 form a network and play diverse roles in regulating Pi signaling and homeostasis in rice.


Self-diffusion in dense granular shear flows.
Diffusivity is a key quantity in describing velocity fluctuations in granular materials. These fluctuations are the basis of many thermodynamic and hydrodynamic models which aim to provide a statistical description of granular systems. We present experimental results on diffusivity in dense, granular shear flows in a two-dimensional Couette geometry. We find that self-diffusivities D are proportional to the local shear rate gamma; with diffusivities along the direction of the mean flow approximately twice as large as those in the perpendicular direction. The magnitude of the diffusivity is D approximately gamma;a(2), where a is the particle radius. However, the gradient in shear rate, coupling to the mean flow, and strong drag at the moving boundary lead to particle displacements that can appear subdiffusive or superdiffusive. In particular, diffusion appears to be superdiffusive along the mean flow direction due to Taylor dispersion effects and subdiffusive along the perpendicular direction due to the gradient in shear rate. The anisotropic force network leads to an additional anisotropy in the diffusivity that is a property of dense systems and has no obvious analog in rapid flows. Specifically, the diffusivity is suppressed along the direction of the strong force network. A simple random walk simulation reproduces the key features of the data, such as the apparent superdiffusive and subdiffusive behavior arising from the mean velocity field, confirming the underlying diffusive motion. The additional anisotropy is not observed in the simulation since the strong force network is not included. Examples of correlated motion, such as transient vortices, and Lévy flights are also observed. Although correlated motion creates velocity fields which are qualitatively different from collisional Brownian motion and can introduce nondiffusive effects, on average the system appears simply diffusive.


A urine inhibitor of interleukin 1 activity affects both interleukin 1 alpha and 1 beta but not tumor necrosis factor alpha.
Urine from monocytic leukemia and other febrile patients contains an inhibitor of interleukin 1 (IL-1), as measured by prostaglandin E2 and collagenase production by human fibroblasts and synovial cells. With the use of recombinant IL-1, the IL-1 inhibitor was partially purified by using ammonium sulfate precipitation, anion-exchange, and gel filtration chromatographies. IL-1 inhibitory activity elutes with an 18,000 to 25,000 apparent molecular size. The same fractions also inhibit IL-1 assayed by the proliferation of murine thymocytes and human fibroblasts. Both forms of human recombinant IL-1, IL-1 alpha and IL-1 beta, which show only 26% homology, but nevertheless bind to the same receptor, are affected by this natural inhibitor to the same extent. In contrast, human recombinant tumor necrosis factor, which shares some of the biologic activities of IL-1, is not inhibited by the urinary IL-1 inhibitor. This study shows that the various biologic activities of both forms of human recombinant IL-1 are inhibited by a partially purified natural urine-derived factor.


Professional self-regulation: peer review.
Dentistry, in fulfilling its responsibility for self-regulation, has established a highly specific and formalized method of peer review. Its primary purpose is to mediate problems between patients, dentists, and third parties relative to the quality of treatment and appropriateness of care. It is a confidential service provided to the public and profession at no cost and accomplished with the voluntary cooperation of all parties involved. Decisions of peer review committees are advisory in nature but offer an alternative to legal resolution of misunderstandings and problems in patient care. The profession, by continually refining peer review and by educating its members and the public to its function, is providing a unique and valuable service to patients, dentists, and third parties. The objective and consistent application of this self-monitoring process is but one mechanism by which dentistry seeks to serve the public and perpetuate its high standard of care.


beta-Catenin and actin reorganization in HGF/SF response of ST14A cells.
Hepatocyte growth factor/scatter factor (HGF/SF) is a pleiotropic factor that activates proliferation, differentiation, and migration of various cell types. Its action is mediated by c-Met, a receptor endowed with tyrosine kinase activity that activates complex signaling cascades and mediates diverse cell responses. Although HGF action was first demonstrated in epithelial cells, expression of HGF and c-Met receptor has also been described in developing and adult mammalian brain. In the developing central nervous system, areas of HGF and c-Met expression are coincident with the migratory pathway of precursor cells. In the present article we report that the interaction between c-Met and HGF/SF in striatal progenitor ST14A cells triggers a signaling cascade that induces modification of cell morphology, with decreased cell-cell interactions and increased cell motility; in particular, we analyzed the reorganization of the actin cytoskeleton and the delocalization of beta-catenin and N-cadherin. The testing of other neurotrophic factors (NGF, BDNF, NT3, and CNTF) showed that the observed modifications were peculiar to HGF. We show that phosphoinositide 3-kinase inhibitor treatment, which blocks cell scattering induced by HGF/SF, does not abolish actin and beta-catenin redistribution. The effects of HGF/SF on primary spinal cord cell cultures were also investigated, and HGF/SF was found to have a possible motogenic effect on these cells. The data reported suggest that HGF could play a role in the early steps of neurogenesis as a motogenic factor.


Clinical profile of children experiencing in-hospital clinical deterioration requiring transfer to a higher level of care.
Few studies have described the various reasons for unplanned transfer to a higher level of care due to clinical deterioration and the clinical profile of those paediatric patients. However, an understanding of the nature of illness is important to patient safety. This study aimed to describe the frequency and clinical characteristics of children who experience unplanned transfer to a higher level of care due to clinical deterioration. A retrospective, descriptive registry study design was used. Of the 92 paediatric patients included, 69% (n = 64) was male. The median age was 2.1 years (interquartile range 0.4-6.9) with 33% being infants under 1 year. The highest number (61.3%) of transfers occurred between 8 and 16 hours. In the 24 hours leading up to a transfer due to clinical deterioration, 15 patients had no vital parameters documented, and 77 patients had least one vital parameter measured. Physiological abnormalities were present in 19 (37.7%) of the 77 patients where vital parameters were documented. This study provides essential baseline data to inform further research to improve care and treatment for critically ill children in paediatric wards. This study's findings suggest reporting of vital parameters is incomplete and infrequent.


Attitudes Toward Fruit and Vegetable Consumption and Farmers' Market Usage Among Low-Income North Carolinians.
Low fruit and vegetable (F&V) consumption is associated with higher rates of obesity and chronic disease among low-income individuals. Understanding attitudes towards F&V consumption and addressing policy and environmental changes could help improve diet and reduce disease risk. A survey of North Carolinians receiving government assistance was used to describe benefits, barriers, and facilitators of eating F&V and shopping at farmers' markets in this population. A total of 341 eligible individuals from 14 counties completed the survey. The most commonly cited barriers to eating F&V were cost (26.4%) and not having time to prepare F&V (7.3%). Facilitators included access to affordable locally grown F&V (13.5%) and knowledge to quickly and easily prepare F&V (13.2%). Among people who did not use farmers' markets, common barriers to shopping there were not being able to use food assistance program benefits (35.3%) and not knowing of a farmers' market in their area (28.8%); common facilitators included transportation (24.8%) and having more information about farmers' market hours (22.9%). In addition to breaking down structural/environmental barriers to farmers' market usage, there is a need to disseminate promotional information about farmers' markets, including hours, location, and accepted forms of payment.


Current application of selective COX-2 inhibitors in cancer prevention and treatment.
The multistep process of carcinogenesis, which can take many years, provides many opportunities for intervention to inhibit disease progression. Effective chemoprevention agents may reduce the risk of cancer by inhibiting the initiation stage of carcinoma through induction of apoptosis or DNA repair in cells harboring mutations, or they may act to prevent promotion of tumor growth. Similarly, chemoprevention may entail blocking cancer progression to an invasive phenotype. Over the past decade, in vitro, preclinical, and clinical data have supported the hypothesis that cyclooxygenase (COX)-2 plays a central role in oncogenesis and that treatment with COX-2 inhibitors offers an effective chemoprevention strategy, as exemplified by the activity of celecoxib (Celebrex) in familial adenomatous polyposis. These COX-2 data have contributed to initiation of clinical trials testing COX-2 inhibitors for the chemoprevention of a wide variety of cancers that overexpress COX-2.


Bacillus halodurans Strain C125 Encodes and Synthesizes Enzymes from Both Known Pathways To Form dUMP Directly from Cytosine Deoxyribonucleotides.
Analysis of the genome of Bacillus halodurans strain C125 indicated that two pathways leading from a cytosine deoxyribonucleotide to dUMP, used for dTMP synthesis, were encoded by the genome of the bacterium. The genes that were responsible, the comEB gene and the dcdB gene, encoding dCMP deaminase and the bifunctional dCTP deaminase:dUTPase (DCD:DUT), respectively, were both shown to be expressed in B. halodurans, and both genes were subject to repression by the nucleosides thymidine and deoxycytidine. The latter nucleoside presumably exerts its repression after deamination by cytidine deaminase. Both comEB and dcdB were cloned, overexpressed in Escherichia coli, and purified to homogeneity. Both enzymes were active and displayed the expected regulatory properties: activation by dCTP for dCMP deaminase and dTTP inhibition for both enzymes. Structurally, the B. halodurans enzyme resembled the Mycobacterium tuberculosis enzyme the most. An investigation of sequenced genomes from other species of the genus Bacillus revealed that not only the genome of B. halodurans but also the genomes of Bacillus pseudofirmus, Bacillus thuringiensis, Bacillus hemicellulosilyticus, Bacillus marmarensis, Bacillus cereus, and Bacillus megaterium encode both the dCMP deaminase and the DCD:DUT enzymes. In addition, eight dcdB homologs from Bacillus species within the genus for which the whole genome has not yet been sequenced were registered in the NCBI Entrez database.


Biochemical and physical parameters of the electrical currents measured with the ADP/ATP carrier by photolysis of caged ADP and ATP.
The transport by the mitochondrial ADP/ATP carrier (AAC) has been shown in a preliminary communication to produce electrical capacitive currents on photolysis of caged ATP or ADP with reconstituted AAC liposomes attached to black lipid membranes [Brustovetsky, N., Becker, A., Klingenberg, M., and Bamberg, E. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 664-668]. Here we study the relation of the currents to ADP/ATP fluxes, the interaction of caged ADP and ATP with AAC and other basic facets of this method. Caged ADP and ATP are not transported by the AAC, as shown in mitochondria. Flux measurements with reconstituted AAC show that caged nucleotides are competitive inhibitors (Ki = 5 microM for caged ADP and 1 microM for caged ATP). Caged ATP competes with photolyzed ATP as shown by the dependence of the currents on the caged ATP concentration and on the light intensity. A competition of added ADP with caged ATP on the currents yields Ki = 50 microM for ADP. We conclude that caged ADP and ATP bind tighter to AAC than ADP or ATP, allowing immediate initiation of translocation by in situ photolysis. The caged compounds bind preferentially at the cytosolic side of AAC. With a regenerative hexokinase + glucose system, the currents are stabilized in repetitive flashes and can be used for applying inhibitors etc. during a flash series. The currents are completely inhibited by the combined addition of the AAC inhibitors bongkrekate (BKA) and carboxyatractylate (CAT). The partial inhibition by CAT or BKA is dependent on the number of flash cycles increasing from 60% to 90%, and by replacing chloride with gluconate from only 30% to 90%. The current are increased by a K+ diffusion potential (valinomycin + KCl) and decreased by the permeant anion TPB-. The pH dependence of the currents and of the parallel flux measurements indicates that only the fully charged ATP4- and ADP3- are transported. A strong temperature dependence of the currents with a break at 15 degrees C (EA = 95 and 28 kJ) agrees with former measurements of flux rates in mitochondria. In conclusion, the capacitive currents faithfully reflect AAC transport function and are a powerful tool for investigating the charge transfer in transport.


Adhesion molecules for mouse primordial germ cells.
In the present article we will focus on the adhesion molecules expressed by mouse primordial germ cells (PGCs) and will discuss the role that they play, or are believed to play, in two crucial processes of PGC development, namely cell lineage specification and migration into the gonadal ridges. Recent findings indicate that the adhesion-dependent allocation of the PGC precursors to a niche within the epiblast and the forming extraembryonic mesoderm during the pre-gastrulation period is crucial for their commitment. Subsequently, PGC migration and homing within the gonadal ridges require integrated signals involving contact of PGCs with extracellular matrix molecules and cellular substrates or repulsion from them, adhesion among PGCs themselves and attraction by the developing gonads. A number of adhesion, or putative adhesion molecules, have been identified in mammalian PGCs, mainly in the mouse. These molecules belong to three adhesion molecule families such as cadherins (E-P- and N-cadherins), integrins and the IgG superfamily (PECAM-1). Moreover oligosaccarides (LewisX) and growth factor receptors (c-Kit) can also play adhesive roles in some stages of PGC development. An understanding of how genes encoding adhesive molecules are regulated in PGCs and the molecular pathways associated with the functions of adhesion receptors is crucial in furthering our knowledge of PGC biology. Adhesion molecules might once again turn out to be crucial in controlling not only the germ cell lineage and PGC migration but also the PGC differentiation fate itself.


Real-time monitoring of enzyme-free strand displacement cascades by colorimetric assays.
The enzyme-free toehold-mediated strand displacement reaction has shown potential for building programmable DNA circuits, biosensors, molecular machines and chemical reaction networks. Here we report a simple colorimetric method using gold nanoparticles as signal generators for the real-time detection of the product of the strand displacement cascade. During the process the assembled gold nanoparticles can be separated, resulting in a color change of the solution. This assay can also be applied in complex mixtures, fetal bovine serum, and to detect single-base mismatches. These results suggest that this method could be of general utility to monitor more complex enzyme-free strand displacement reaction-based programmable systems or for further low-cost diagnostic applications.


Efficacy and safety of hepcidin-based screen-and-treat approaches using two different doses versus a standard universal approach of iron supplementation in young children in rural Gambia: a double-blind randomised controlled trial.
Iron deficiency prevalence rates frequently exceed 50 % in young children in low-income countries. The World Health Organization (WHO) recommended universal supplementation of young children where anaemia rates are >40 %. However, large randomized trials have revealed that provision of iron to young children caused serious adverse effects because iron powerfully promotes microbial growth. Hepcidin - the master regulator of iron metabolism that integrates signals of infection and iron deficiency - offers the possibility of new solutions to diagnose and combat global iron deficiency. We aim to evaluate a hepcidin-screening-based iron supplementation intervention using hepcidin cut-offs designed to indicate that an individual requires iron, is safe to receive it and will absorb it. The study is a proof-of-concept, three-arm, double blind, randomised controlled, prospective, parallel-group non-inferiority trial. Children will be randomised to receive, for a duration of 12 weeks, one of three multiple micronutrient powders (MNP) containing: A) 12 mg iron daily; B) 12 mg or 0 mg iron daily based on a weekly hepcidin screening indicating if iron can be given for the next seven days or not; C) 6 mg or 0 mg iron daily based on a weekly hepcidin screening indicating if iron can be given for the next seven days or not. The inclusion criteria are age 6-23 months, haemoglobin (Hb) concentration between 7 and 11 g/dL, z-scores for Height-for-Age, Weight-for-Age and Weight-for-Height > -3 SD and free of malaria. Hb concentration at 12 weeks will be used to test whether the screen-and-treat approaches are non-inferior to universal supplementation. Safety will be assessed using caregiver reports of infections, in vitro bacterial and P. falciparum growth assays and by determining the changes in the gut microbiota during the study period. A screen-and-treat approach using hepcidin has the potential to make iron administration safer in areas with widespread infections. If this proof-of-concept study shows promising results the development of a point-of-care diagnostic test will be the next step. ISRCTN07210906 , 07/16/2014.


Multi-tissue epigenetic analysis identifies distinct associations underlying insulin resistance and Alzheimer's disease at CPT1A locus.
Insulin resistance (IR) is a major risk factor for Alzheimer's disease (AD) dementia. The mechanisms by which IR predisposes to AD are not well-understood. Epigenetic studies may help identify molecular signatures of IR associated with AD, thus improving our understanding of the biological and regulatory mechanisms linking IR and AD. We conducted an epigenome-wide association study of IR, quantified using the homeostatic model assessment of IR (HOMA-IR) and adjusted for body mass index, in 3,167 participants from the Framingham Heart Study (FHS) without type 2 diabetes at the time of blood draw used for methylation measurement. We identified DNA methylation markers associated with IR at the genome-wide level accounting for multiple testing (P < 1.1 × 10-7) and evaluated their association with neurological traits in participants from the FHS (N = 3040) and the Religious Orders Study/Memory and Aging Project (ROSMAP, N = 707). DNA methylation profiles were measured in blood (FHS) or dorsolateral prefrontal cortex (ROSMAP) using the Illumina HumanMethylation450 BeadChip. Linear regressions (ROSMAP) or mixed-effects models accounting for familial relatedness (FHS) adjusted for age, sex, cohort, self-reported race, batch, and cell type proportions were used to assess associations between DNA methylation and neurological traits accounting for multiple testing. We confirmed the strong association of blood DNA methylation with IR at three loci (cg17901584-DHCR24, cg17058475-CPT1A, cg00574958-CPT1A, and cg06500161-ABCG1). In FHS, higher levels of blood DNA methylation at cg00574958 and cg17058475 were both associated with lower IR (P = 2.4 × 10-11 and P = 9.0 × 10-8), larger total brain volumes (P = 0.03 and P = 9.7 × 10-4), and smaller log lateral ventricular volumes (P = 0.07 and P = 0.03). In ROSMAP, higher levels of brain DNA methylation at the same two CPT1A markers were associated with greater risk of cognitive impairment (P = 0.005 and P = 0.02) and higher AD-related indices (CERAD score: P = 5 × 10-4 and 0.001; Braak stage: P = 0.004 and P = 0.01). Our results suggest potentially distinct epigenetic regulatory mechanisms between peripheral blood and dorsolateral prefrontal cortex tissues underlying IR and AD at CPT1A locus.


Transcriptomics and Spatial Proteomics for Discovery and Validation of Missing Proteins in the Human Ovary.
Efforts to understand the complexities of human biology encompass multidimensional aspects, with proteins emerging as crucial components. However, studying the human ovary introduces unique challenges due to its complex dynamics and changes over a lifetime, varied cellular composition, and limited sample access. Here, four new RNA-seq samples of ovarian cortex spanning ages of 7 to 32 were sequenced and added to the existing data in the Human Protein Atlas (HPA) database www.proteinatlas.org, opening the doors to unique possibilities for exploration of oocyte-specific proteins. Based on transcriptomics analysis of the four new tissue samples representing both prepubertal girls and women of fertile age, we selected 20 protein candidates that lacked previous evidence at the protein level, so-called "missing proteins" (MPs). The proteins were validated using high-resolution antibody-based profiling and single-cell transcriptomics. Fourteen proteins exhibited consistent single-cell expression patterns in oocytes and granulosa cells, confirming their presence in the ovary and suggesting that these proteins play important roles in ovarian function, thus proposing that these 14 proteins should no longer be classified as MPs. This research significantly advances the understanding of MPs, unearthing fresh avenues for prospective exploration. By integrating innovative methodologies and leveraging the wealth of data in the HPA database, these insights contribute to refining our understanding of protein roles within the human ovary and opening the doors for further investigations into missing proteins and human reproduction.


Rapid actions of xenoestrogens disrupt normal estrogenic signaling.
Some chemicals used in consumer products or manufacturing (e.g. plastics, surfactants, pesticides, resins) have estrogenic activities; these xenoestrogens (XEs) chemically resemble physiological estrogens and are one of the major categories of synthesized compounds that disrupt endocrine actions. Potent rapid actions of XEs via nongenomic mechanisms contribute significantly to their disruptive effects on functional endpoints (e.g. cell proliferation/death, transport, peptide release). Membrane-initiated hormonal signaling in our pituitary cell model is predominantly driven by mERα with mERβ and GPR30 participation. We visualized ERα on plasma membranes using many techniques in the past (impeded ligands, antibodies to ERα) and now add observations of epitope proximity with other membrane signaling proteins. We have demonstrated a range of rapid signals/protein activations by XEs including: calcium channels, cAMP/PKA, MAPKs, G proteins, caspases, and transcription factors. XEs can cause disruptions of the oscillating temporal patterns of nongenomic signaling elicited by endogenous estrogens. Concentration effects of XEs are nonmonotonic (a trait shared with natural hormones), making it difficult to design efficient (single concentration) toxicology tests to monitor their harmful effects. A plastics monomer, bisphenol A, modified by waste treatment (chlorination) and other processes causes dephosphorylation of extracellular-regulated kinases, in contrast to having no effects as it does in genomic signaling. Mixtures of XEs, commonly found in contaminated environments, disrupt the signaling actions of physiological estrogens even more severely than do single XEs. Understanding the features of XEs that drive these disruptive mechanisms will allow us to redesign useful chemicals that exclude estrogenic or anti-estrogenic activities.


Optimal unified combination rule in application of Dempster-Shafer theory to lung cancer radiotherapy dose response outcome analysis.
Our previous study demonstrated the application of the Dempster-Shafer theory of evidence to dose/volume/outcome data analysis. Specifically, it provided Yager's rule to fuse data from different institutions pertaining to radiotherapy pneumonitis versus mean lung dose. The present work is a follow-on study that employs the optimal unified combination rule, which optimizes data similarity among independent sources. Specifically, we construct belief and plausibility functions on the lung cancer radiotherapy dose outcome datasets, and then apply the optimal unified combination rule to obtain combined belief and plausibility, which bound the probabilities of pneumonitis incidence. To estimate the incidence of pneumonitis at any value of mean lung dose, we use the Lyman-Kutcher-Burman (LKB) model to fit the combined belief and plausibility curves. The results show that the optimal unified combination rule yields a narrower uncertainty range (as represented by the belief-plausibility range) than Yager's rule, which is also theoretically proven.


[Cancer of the penis. Personal experience].
The author reports 123 cases of penile cancer, with a follow up of fifteen years, and an incidence of 38.2% in North East Argentina. He considers the clinical and diagnostic aspects, and proposes a classification with an eye to suitable treatment, based on detailed results. He stresses the need for early diagnosis, and the importance of preventive treatment. This article does not claim to be an exhaustive study of this pathology, but it does recount the author's personal experience, based on exhaustive and detailed analysis of a large number of cases which highlight the frequency of this type of neoplasia in North Eastern Argentina, and which give rise to a considerable scientific concern. Since 1968, he has examined and treated 123 cases of penile cancer. This experience has provided an interesting series of statistics concerning diagnosis, management, prognosis and prophylaxis.


[Effect of double helix conformation transitions on DNA hypochromism].
The sensitivity of the first UV-absorption band hypochromism of DNA to changes in double helix form has been investigated theoretically. The hypochromism of DNA is studied within the perturbation theory and the nearest neighbour approximation. The heterogeneity of DNA is taken into account by introducing the nearest neighbour frequencies. The electronic characteristics of monomers (nucleotide bases) are calculated by the quantum mechanical PPP CI method. The hypochromism of A-, B-, C-, D- and T-forms of DNA is calculated. A slight dependence of the hypochromism on double helix conformation is obtained. This result is in agreement with experimental data. The reasons for the observed non-sensitivity of the DNA UV-absorption spectrum to conformational transitions within a two-stranded structure are explained. It is shown that the theory leads to an agreement with experiments because a large number of electronic transitions in nucleotide bases and the heterogeneity of DNA are taken into account.


Retinoic acid receptor-alpha is stabilized in a repressive state by its C-terminal, isotype-specific F domain.
Retinoic acid receptors (RARs) are hormone-regulated transcription factors that play multiple roles in vertebrate development and differentiation. Three isotypes of RARs, alpha, beta, and gamma, are encoded by distinct genetic loci and possess distinct transcriptional properties. Typically, RARalpha represses target gene transcription in the absence of hormone, whereas RARbeta and gamma fail to repress under these conditions. This inability of RARbeta and RARgamma to repress transcription is due to intramolecular interactions between helix 3 and helix 12 of the hormone binding domains of these isotypes that inhibit corepressor binding while favoring coactivator binding. We report here that the converse ability of RARalpha to repress requires the integrity of the receptor F domain, a domain that maps C-terminal to helix 12, varies in sequence among different nuclear receptors, and is of poorly understood function. The F domain appears to help stabilize helix 12 of RARalpha in a more open position that enhances corepressor binding and inhibits coactivator binding in the absence of hormone. Intriguingly, the RARalpha F domain is isotype autonomous in its function. We speculate that the RARalpha F domain may dock elsewhere on the receptor surface, and this intramolecular interaction may maintain RARalpha helix 12 in an open, repression-competent conformation.


Apparent affinity of the Na/K pump for ouabain in cultured chick cardiac myocytes. Effects of Nai and Ko.
The measured apparent affinity (K0.5) of the Na/K pump for ouabain has been reported to vary over a wide range. In a previous report we found that changing Nai could alter apparent affinity by at least an order of magnitude and that the model presented predicted this variability. To increase our understanding of this variability, isolated cells or two- to three-cell clusters of cardiac myocytes from 11-d embryonic chick were used to measure the effects of Nai and Ko on the K0.5 of the Na/K pump for ouabain. Myocytes were whole-cell patch clamped and Na/K pump current (Ip) was measured in preparations exposed to a Ca-free modified Hank's solution (HBSS) that contained 1 mM Ba, 10 mM Cs, and 0.1 mM Cd. Under these conditions there are no Ko-sensitive currents other than Ip because removal of Ko in the presence of ouabain had no effect on the current-voltage (I-V) relation. The I-V relation for Ip showed that in the presence of 5.4 mM Ko and 51 mM Nai, Ip has a slight voltage dependence, decreasing approximately 30% from 0 to -130 mV. Increasing Nai in the patch pipette from 6 to 51 mM (Ko = 5.4 mM) caused Ip to increase from 0.46 +/- 0.07 (n = 5) to 1.34 +/- 0.08 microA/cm2 (n = 13) with a K0.5 for Nai of 17.4 mM and decreased the K0.5 for ouabain from 18.5 +/- 1.8 (n = 4) to 3.1 +/- 0.4 microM (n = 3). Similarly, varying Ko between 0.3 and 10.8 mM (Nai = 24 mM) increased Ip from 0.13 +/- 0.01 (n = 5) to 0.90 +/- 0.05 microA/cm2 (n = 5) with a K0.5 for Ko of 1.94 mM and increased K0.5 for ouabain from 0.56 +/- 0.14 (n = 3-6) to 10.0 +/- 1.1 microM (n = 6). All of these changes are predicted by the model presented. A qualitative explanation of these results is that Nai and Ko interact with the Na/K pump to shift the steady-state distribution of the Na/K pump molecules among the kinetic states. This shift in state distribution alters the probability that the Na/K pump will be in the conformation that binds ouabain with high affinity, thus altering the apparent affinity. In intact cells, the measured apparent affinity represents a combination of all the rate constants in the model and does not equate to simple first-order binding kinetics.(ABSTRACT TRUNCATED AT 400 WORDS)


Effect of highly active antiretroviral therapy and thymic transplantation on immunoreconstitution in HIV infection.
The purpose of this study was to determine whether thymic transplantation in addition to highly active antiretroviral therapy (HAART) will restore T cell function in HIV infection. Eight treatment-naive HIV-infected patients with CD4+ T cell counts of 200-500/mm3 were randomized into thymic transplantation and control arms. All patients received HAART (zidovudine, lamivudine, and ritonavir) for 6 weeks prior to transplantation. Thymic transplantation was done without immunosuppression, using postnatal HLA-unmatched cultured allogeneic thymus tissue. Patients were immunized every 6 months with the neoantigen keyhole limpet hemocyanin (KLH) and the recall antigen tetanus toxoid (TT). T cell phenotype and function and T cell receptor rearrangement excision circles (TRECs) were assessed. Thymic allografts were biopsied at 2 months. Six HIV-infected patients completed the study. Four patients received cultured allogeneic postnatal thymic grafts, two others were controls. CD4+ T cell counts increased and T cell-proliferative responses to Candida antigen and TT normalized in all patients. Proliferative responses to KLH developed in three of four transplant recipients and one of two controls. Patients responding to KLH after secondary immunization had greater TREC increases compared with the patients who did not respond. All thymic allografts were rejected within 2 months. In summary, four of six patients developed T cell-proliferative responses to the neoantigen KLH over the first 2 years of HAART. The transplanted thymus tissue, however, was rejected. There was no clear difference in restoration of T cell function in the transplant recipients compared with the controls. Increases in TRECs after initiation of HAART may correlate with improved immune function.


From new frontiers to new standards of practice: advances in radiotherapy planning and delivery.
Radiation therapy treatment planning and delivery capabilities have changed dramatically since the introduction of three-dimensional treatment planning in the 1980s and continue to change in response to the implementation of new technologies. CT simulation and three-dimensional radiation treatment planning systems have become the standard of practice in clinics around the world. Medical accelerator manufacturers have employed advanced computer technology to produce treatment planning/delivery systems capable of precise shaping of dose distributions via computer-controlled multileaf collimators, in which the beam fluence is varied optimally to achieve the plan prescription. This mode of therapy is referred to as intensity-modulated radiation therapy (IMRT), and is capable of generating extremely conformal dose distributions including concave isodose volumes that provide conformal target volume coverage and avoidance of specific sensitive normal structures. IMRT is rapidly being implemented in clinics throughout the USA. This increasing use of IMRT has focused attention on the need to better account for both intrafraction and interfraction spatial uncertainties, which has helped spur the development of treatment machines with integrated planar and volumetric advanced imaging capabilities. In addition, advances in both anatomical and functional imaging provide improved ability to define the tumor volumes. Advances in all these technologies are occurring at a record pace and again pushing the cutting-edge frontiers of radiation oncology from IMRT to what is now referred to as image-guided IMRT, or simply image-guided radiation therapy (IGRT). A brief overview is presented of these latest advancements in conformal treatment planning and treatment delivery.


Mucus Plugs in Asthma at CT Associated with Regional Ventilation Defects at 3He MRI.
Background Airway mucus plugs in asthma are associated with exacerbation frequency, increased eosinophilia, and reduced lung function. The relationship between mucus plugs and spatially overlapping ventilation abnormalities observed at hyperpolarized gas MRI has not been assessed quantitatively. Purpose To assess regional associations between CT mucus plugs scored by individual bronchopulmonary segment and corresponding measurements of segmental ventilation defect percentage (VDP) at hyperpolarized helium 3 (3He) MRI. Materials and Methods In this secondary analysis of a Health Insurance Portability and Accountability Act-compliant prospective observational cohort, participants in the Severe Asthma Research Program (SARP) III (NCT01760915) between December 2012 and August 2015 underwent hyperpolarized 3He MRI to determine segmental VDP. Segmental mucus plugs at CT were scored by two readers, with segments scored as plugged only if both readers agreed independently. A linear mixed-effects model controlling for interpatient variability was then used to assess differences in VDP in plugged versus plug-free segments. Results Forty-four participants with asthma were assessed (mean age ± standard deviation, 47 years ± 15; 29 women): 19 with mild-to-moderate asthma and 25 with severe asthma. Mucus plugs were observed in 49 total bronchopulmonary segments across eight of 44 patients. Segments containing mucus plugs had a median segmental VDP of 25.9% (25th-75th percentile, 7.3%-38.3%) versus 1.4% (25th-75th percentile, 0.1%-5.2%; P < .001) in plug-free segments. Similarly, the model estimated a segmental VDP of 18.9% (95% CI: 15.7, 22.2) for mucus-plugged segments versus 5.1% (95% CI: 3.3, 7.0) for plug-free segments (P < .001). Participants with one or more mucus plugs had a median whole-lung VDP of 11.1% (25th-75th percentile, 7.1%-18.9%) versus 3.1% (25th-75th percentile, 1.1%-4.4%) in those without plugs (P < .001). Conclusion Airway mucus plugging at CT was associated with reduced ventilation in the same bronchopulmonary segment at hyperpolarized helium 3 MRI, suggesting that mucus plugging may be an important cause of ventilation defects in asthma. © RSNA, 2021 Online supplemental material is available for this article.


Resurfacing hip replacement and cemented total hip replacement have equivalent outcome at one year in a disease matched population: a case-control study of patient reported outcome measures.
Resurfacing hip replacement has demonstrated good survival and outcomes for cohorts of younger male patients, but few controlled studies exist. In this study we compared patient reported outcome measures and satisfaction scores at one year following resurfacing hip replacement in 69 male patients with two control groups of equal numbers undergoing cemented total hip replacement: aged-matched patients and disease matched patients. At one year we found no difference in improvement in patient reported outcome measures between patients undergoing resurfacing hip replacement and disease matched patients, whereas patients undergoing resurfacing hip replacement had a statistically significant improvement in Oxford Hip Score compared to the age-matched controls (p<0.047) although this was below the minimally clinically detectable difference. Resurfacing hip replacement and total hip replacement both confer increase in patient reported outcome scores and high patient satisfaction at one year. The results of this study will allow better counselling of patients and help inform treatment decisions.


Evidence for diversifying selection of genetic regions of encoding putative collagen-like host-adhesive fibers in Pasteuria penetrans.
Pasteuria spp. belong to a group of genetically diverse endospore-forming bacteria (phylum: Firmicutes) that are known to parasitize plant-parasitic nematodes and water fleas (Daphnia spp.). Collagen-like fibres form the nap on the surface of endospores and the genes encoding these sequences have been hypothesised to be involved in the adhesion of the endospores of Pasteuria spp. to their hosts. We report a group of 17 unique collagen-like genes putatively encoded by Pasteuria penetrans (strain: Res148) that formed five different phylogenetic clusters and suggest that collagen-like proteins are an important source of genetic diversity in animal pathogenic Firmicutes including Pasteuria. Additionally, and unexpectedly, we identified a putative collagen-like sequence which had a very different sequence structure to the other collagen-like proteins but was similar to the protein sequences in Megaviruses that are involved in host-parasite interactions. We, therefore, suggest that these diverse endospore surface proteins in Pasteuria are involved in biological functions, such as cellular adhesion; however, they are not of monophyletic origin and were possibly obtained de novo by mutation or possibly through selection acting upon several historic horizontal gene transfer events.


Salivary signals of European corn borer induce indirect defenses in tomato.
Plants can recognize the insect elicitors and activate its defense mechanisms. European Corn Borer (ECB; Ostrinia nubilalis) saliva, produced from the labial salivary glands and released through the spinneret, is responsible for inducing direct defenses in host plants. Glucose oxidase (GOX) present in the ECB saliva induced direct defenses in tomato. By contrast, GOX activity in ECB saliva was insufficient to trigger defenses in maize, suggesting that host-specific salivary elicitors are responsible for inducing direct defenses in host plants. Our current study further examined whether ECB saliva can trigger indirect defenses in tomato. Relative expression levels of TERPENE SYNTHASE5 (TPS5) and HYDROPEROXIDE LYASE (HPL), marker for indirect defenses in host plants, were monitored. Quantitative real-time PCR analysis revealed that ECB saliva can induce the expression of TPS5 and HPL, suggesting that salivary signals can induce indirect defenses in addition to the direct defenses. Further experiments are required to identify different ECB elicitors that are responsible for inducing direct and indirect defenses in host plants.


Energetic costs of ectoparasite infection in Atlantic salmon.
Parasites are widespread in nature, where they affect the energy budget of hosts, and depending on the imposed pathogenic severity, this may reduce host fitness. However, the energetic costs of parasite infections are rarely quantified. In this study, we measured metabolic rates in recently seawater adapted Atlantic salmon (Salmo salar) infected with the ectoparasitic copepod Lepeophtheirus salmonis and used an aerobic scope framework to assess the potential ecological impact of this parasite-host interaction. The early chalimus stages of L. salmonis did not affect either standard or maximum metabolic rates. However, the later mobile pre-adult stages caused an increase in both standard and maximum metabolic rate yielding a preserved aerobic scope. Notably, standard metabolic rates were elevated by 26%, presumably caused by increased osmoregulatory burdens and costs of mobilizing immune responses. The positive impact on maximum metabolic rates was unexpected and suggests that fish are able to transiently overcompensate energy production to endure the burden of parasites and thus allow for continuation of normal activities. However, infected fish are known to suffer reduced growth, and this suggests that a trade-off exists in acquisition and assimilation of resources despite an uncompromised aerobic scope. As such, when assessing impacts of environmental or biotic factors, we suggest that elevated routine costs may be a stronger predictor of reduced fitness than the available aerobic scope. Furthermore, studying the effects on parasitized fish in an ecophysiological context deserves more attention, especially considering interacting effects of other stressors in the Anthropocene.


Finding our way: On the sharing and reuse of animal telemetry data in Australasia.
The presence and movements of organisms both reflect and influence the distribution of ecological resources in space and time. The monitoring of animal movement by telemetry devices is being increasingly used to inform management of marine, freshwater and terrestrial ecosystems. Here, we brought together academics, and environmental managers to determine the extent of animal movement research in the Australasian region, and assess the opportunities and challenges in the sharing and reuse of these data. This working group was formed under the Australian Centre for Ecological Analysis and Synthesis (ACEAS), whose overall aim was to facilitate trans-organisational and transdisciplinary synthesis. We discovered that between 2000 and 2012 at least 501 peer-reviewed scientific papers were published that report animal location data collected by telemetry devices from within the Australasian region. Collectively, this involved the capture and electronic tagging of 12 656 animals. The majority of studies were undertaken to address specific management questions; rarely were these data used beyond their original intent. We estimate that approximately half (~500) of all animal telemetry projects undertaken remained unpublished, a similar proportion were not discoverable via online resources, and less than 8.8% of all animals tagged and tracked had their data stored in a discoverable and accessible manner. Animal telemetry data contain a wealth of information about how animals and species interact with each other and the landscapes they inhabit. These data are expensive and difficult to collect and can reduce survivorship of the tagged individuals, which implies an ethical obligation to make the data available to the scientific community. This is the first study to quantify the gap between telemetry devices placed on animals and findings/data published, and presents methods for improvement. Instigation of these strategies will enhance the cost-effectiveness of the research and maximise its impact on the management of natural resources.


Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection.
Yersinia pestis is a Gram-negative bacterium that is the causative agent of plague and is widely recognized as a potential biological weapon. Due to the high fatality rate of plague when diagnosis is delayed, the development of rapid, sensitive, specific, and cost-effective methods is needed for its diagnosis. The Y. pestis low calcium response V (LcrV) protein has been identified as a potential microbial biomarker for the diagnosis of plague. In this paper, we present a highly sensitive, paper-based, vertical flow immunoassay (VFI) prototype for the detection of LcrV and the diagnosis of plague. An antigen-capture assay using monoclonal antibodies is employed to capture and detect the LcrV protein, using a colorimetric approach. In addition, the effect of miniaturizing the VFI device is explored based on two different sizes of VFI platforms, denoted as "large VFI" and "mini VFI." Also, a comparative analysis is performed between the VFI platform and a lateral flow immunoassay (LFI) platform to exhibit the improved assay sensitivity suitable for point-of-care (POC) diagnostics. The analytical sensitivity or limit of detection (LOD) in the mini VFI is approximately 0.025 ng/mL, that is, 10 times better than that of the large VFI platform or 80 times over a standard lateral flow configuration. The low LOD of the LcrV VFI appears to be highly suitable for testing clinical samples and potentially diagnosing plague at earlier time points. In addition, optimization of the gold nanoparticle (AuNP) concentration, nanomaterial plasmonic properties, and flow velocity analysis could improve the performance of the VFI. Furthermore, we developed automated image analysis software that shows potential for integrating the diagnostic system into a smartphone. These methods and findings demonstrate that the VFI platform is a highly sensitive device for detecting the LcrV and potentially many other biomarkers.


Fabrication of HKUST-1/ZnO/SA nanocomposite for Doxycycline and Naproxen adsorption from contaminated water.
Doxycycline and Naproxen are among the most widely used drugs in the therapy of CoVID 19 disease found in surface water. Water scarcity in recent years has led to research to treat polluted water. One of the easy and low-cost methods for treatment is adsorption. The utilize of Metal-Organic Frameworks (MOFs) to evacuate pharmaceutical contaminants from water sources has been considered by researchers in the last decade. In this research, HKUST-1/ZnO/SA composite with high adsorption capacity, chemical and water stability, recovery, and reuse properties has been synthesized and investigated. By adding 10 wt% of ZnO and 50 wt% of sodium alginate to HKUST-1, at 25 °C and pH = 7, the specific surface area is reduced by 60%. The parameters of drugs concentration C0 =(5,80) mg/L, time=(15,240) min, and pH= (2,12) were investigated, and the results showed that the HKUST-1/ZnO/SA is stable in water for 14 days and it can be used in 10 cycles with 80% removal efficiency. The maximum Adsorption loading of doxycycline and Naproxen upon HKUST-1/ZnO/SA is 97.58 and 80.04 mg/g, respectively. Based on the correlation coefficient (R2), the pseudo-second-order and the Langmuir isotherm models were selected for drug adsorption. The proposed mechanism of drug uptake is by MOFs, hydrogen bonding, electrostatic bonding, and acid-base interaction.


Inhibition of porcine detrusor contractility by the flavonoid fraction of Bryophyllum pinnatum--a potential phytotherapeutic drug for the treatment of the overactive bladder syndrome.
To determine if the phytotherapeutic agent, Bryophyllum pinnatum, could serve as an alternative drug for the overactive bladder syndrome, and to characterise the fraction responsible for the inhibition of detrusor contractility. Fractions were prepared from the MeOH extract of B. pinnatum and further analysed by HPLC-PDA-MS. Detrusor muscle strips were prepared from porcine bladders and the electrically induced muscle contractility measured by organ bath. The effect of B. pinnatum leaf press juice (2.5-10%), a flavonoid fraction (0.1-1 mg/ml), and a bufadienolide fraction (0.1-40 μg/ml) on detrusor contractility was assessed and compared with controls (polar fraction (0.5-5 mg/ml) and oxybutynin (10(-8)-10(-6) M)). The press juice, at a concentration of 10% led to a reduction of detrusor contractility. Bladder strips treated with the flavonoid fraction showed a significant reduction of the contractility to 21.3 ± 5.2% (1 mg/ml) while the bufadienolide fraction had no inhibitory effect in the investigated concentrations. The polar fraction showed a reduction of the contractility in a pH-dependent fashion. At 10(-6) M concentration oxybutynin reduced the detrusor contractility to 21.9 ± 4.7%. The flavonoid fraction of Bryophyllum pinnatum reduces the porcine detrusor contractility in a dose- and time-dependent manner. Fractions from B. pinnatum may be a new pharmacological approach for the treatment of OAB.


Adaptation of Patterns of Motile Filaments under Dynamic Boundary Conditions.
Boundary conditions are important for pattern formation in active matter. However, it is still not well-understood how alterations in the boundary conditions (dynamic boundary conditions) impact pattern formation. To elucidate the effect of dynamic boundary conditions on the pattern formation by active matter, we investigate an in vitro gliding assay of microtubules on a deformable soft substrate. The dynamic boundary conditions were realized by applying mechanical stress through stretching and compression of the substrate during the gliding assay. A single cycle of stretch-and-compression (relaxation) of the substrate induces perpendicular alignment of microtubules relative to the stretch axis, whereas repeated cycles resulted in zigzag patterns of microtubules. Our model shows that the orientation angles of microtubules correspond to the direction to attain smooth movement without buckling, which is further amplified by the collective migration of the microtubules. Our results provide an insight into understanding the rich dynamics in self-organization arising in active matter subjected to time-dependent boundary conditions.


Intrathecal bupivacaine in humans: influence of volume and baricity of solutions.
The effects of volume and baricity of spinal bupivacaine on block onset, height, duration, and hemodynamics were studied. Ninety patients undergoing endoscopic urologic procedures were randomized to receive 10 mg of intrathecal bupivacaine at L2-L3 level in sitting position. In the operating room, commercial products were diluted as needed with NaCl 0.9% to obtain isobaric solutions (density, 1.005-1.008) or with NaC 10.9% and glucose 30% to obtain hyperbaric solutions (density, 1.031-1.037) of 2, 5, or 10 ml (six groups of 15 patients each). Three minutes after spinal injection the patients were placed in lithotomy position. Sensory blockade was assessed using pinprick and cold sensation tests, and motor blockade was assessed using a four-point scale. Onset times to maximal cephalad spread of spinal blockade were similar with isobaric and hyperbaric solutions. A greater maximal cephalad spread of anesthesia was obtained with diluted isobaric bupivacaine but was not associated with more hypotension. Volume had no effect on cephalad extent of anesthesia with hyperbaric bupivacaine. Times for regression of anesthesia to L2 and offset of motor block were longer with isobaric than with hyperbaric solutions of bupivacaine. The intensity of motor blockade was decreased with diluted hyperbaric bupivacaine. No patient reported back pain. In this study, volume had no significant influence on either cephalad spread or duration of sensory blockade for either isobaric or hyperbaric bupivacaine. Time for offset of anesthesia was shorter with hyperbaric bupivacaine compared with isobaric solutions.


Balkan nephropathy: evolution of our knowledge.
Balkan endemic nephropathy (BEN), originally described in the late 1950s as a chronic tubulointerstitial kidney disease, is identified by its unique epidemiological features. The most remarkable characteristic of BEN is the focal topographical nature that characterizes its occurrence at the global, national, and even household level. BEN affects only certain endemic rural foci along tributaries of the Danube River in the Balkan countries of Bosnia, Bulgaria, Croatia, Romania, and Serbia. The spatial distribution has remained astonishingly unchanged with time because the disease affects the same endemic clusters as 50 years ago. The natural course of the disease is characterized by universal development of end-stage renal disease and the frequent development of upper urinary tract tumors, posing a substantial disease burden to the afflicted areas. The greatest challenge in the study of BEN has been the elucidation of its cause. The unique features of the disease, in particular its endemic nature and the long incubation period required for the disease to develop, have led to the proposal that BEN represents a unique environmental disease. The quest for the responsible environmental factor has been long and diverse, and although no definitive answer has been provided to date, converging lines of evidence support the theory that long-term consumption of food contaminated with aristolochic acid underlies the pathogenesis of BEN. The present review describes the evolution of our knowledge of BEN in relation to the development of the main theories for its pathogenesis.


Multilevel versus single-level regression for the analysis of multilevel information: The case of quantitative intersectional analysis.
Intersectional MAIHDA involves applying multilevel models in order to estimate intercategorical inequalities. The approach has been validated thus far using both simulations and empirical applications, and has numerous methodological and theoretical advantages over single-level approaches, including parsimony and reliability for analyzing high-dimensional interactions. In this issue of SSM, Lizotte, Mahendran, Churchill and Bauer (hereafter "LMCB") assert that there has been insufficient clarity on the interpretation of fixed effects regression coefficients in intersectional MAIHDA, and that stratum-level residuals in intersectional MAIHDA are not interpretable as interaction effects. We disagree with their second assertion; however, the authors are right to call for greater clarity. For this purpose, in this response we have three main objectives. (1) In their commentary, LMCB incorrectly describe model predictions based on MAIHDA fixed effects as estimates of "grand means" (or the mean of means), when they are actually "precision-weighted grand means." We clarify the differences between average predicted values obtained by different models, and argue that predictions obtained by MAIHDA are more suitable to serve as reference points for residual/interaction effects. This further enables us to clarify the interpretation of residual/interaction effects in MAIHDA and conventional models. Using simple simulations, we demonstrate conditions under which the precision-weighted grand mean resembles a grand mean, and when it resembles a population mean (or the mean of all individual observations) obtained using single-level regression, explaining the results obtained by LMCB and informing future research. (2) We construct a modification to MAIHDA that constrains the fixed effects so that the resulting model predictions provide estimates of population means, which we use to demonstrate the robustness of results reported by Evans et al. (2018). We find that stratum-specific residuals obtained using the two approaches are highly correlated (Pearson corr = 0.98, p < 0.0001) and no substantive conclusions would have been affected if the preference had been for estimating population means. However, we advise researchers to use the original, unconstrained MAIHDA. (3) Finally, we outline the extent to which single-level and MAIHDA approaches address the fundamental goals of quantitative intersectional analyses and conclude that intersectional MAIHDA remains a promising new approach for the examination of inequalities.


Current Controversies in Thrombolytic Use in Acute Pulmonary Embolism.
Acute pulmonary embolism (PE) has an annual incidence of 100,000 cases in the United States and is divided into three categories: nonmassive, submassive, and massive. Several studies have evaluated the use of thrombolytics in submassive and massive PE. Our aim was to provide emergency physicians with an updated review of the controversy about the use of thrombolytics in submassive and massive PE. Nonmassive PE is defined as PE in the setting of no signs of right ventricular strain (echocardiogram or biomarker) and hemodynamic stability. Submassive PE is defined as evidence of right ventricular strain with lack of hemodynamic instability. Massive PE occurs with occlusive thromboembolism that causes hemodynamic instability. Thrombolysis is warranted in patients with massive PE. Thrombolytic use in submassive PE with signs of right ventricular strain or damage presents a quandary for physicians. Several recent studies have evaluated the use of thrombolytics in patients with submassive PE. These studies have inconsistent definitions of submassive PE, evaluate differing primary outcomes, and use different treatment protocols with thrombolytics and anticoagulation agents. Although significant study heterogeneity exists, thrombolytics can improve long-term outcomes, with decreased bleeding risk with half-dose thrombolytics and catheter-directed treatments. Major bleeding significantly increases in patients over age 65 years. The risks and benefits of thrombolytic treatment-primarily improved long-term outcomes-should be considered on a case-by-case basis. Shared decision-making with the patient discussing the risks and benefits of treatment is advised. Thrombolytic use in massive PE is warranted, but patients with submassive PE require case-by-case analysis with shared decision making. The risks, including major hemorrhage, and benefits, primarily improved long-term outcomes, should be considered. Half-dose thrombolytics and catheter-directed treatment demonstrate advantages with decreased risk of bleeding and improved long-term functional outcomes. Further studies that assess risk stratification, functional outcomes, and treatment protocols are needed.


Study methods, recruitment, sociodemographic findings, and demographic representativeness in the OPPERA study.
This paper describes methods used in the project "Orofacial Pain Prospective Evaluation and Risk Assessment" (OPPERA) and evaluates sociodemographic characteristics associated with temporomandibular disorders (TMD) in the OPPERA case-control study. Representativeness was investigated by comparing sociodemographic profiles of OPPERA participants with population census profiles of counties near study sites and by comparing age and gender associations with TMD in OPPERA and the 2007 to 2009 US National Health Interview Survey. Volunteers aged 18 to 44 years were recruited at 4 US study sites: 3,263 people without TMD were enrolled into the prospective cohort study; 1,633 of them were selected as controls for the baseline case-control study. Cases were 185 volunteers with examiner-classified TMD. Distributions of some demographic characteristics among OPPERA participants differed from census profiles, although there was less difference in socioeconomic profiles. Odds of TMD was associated with greater age in this 18 to 44 year range; females had 3 times the odds of TMD as males; and relative to non-Hispanic-Whites, other racial groups had one-fifth the odds of TMD. Age and gender associations with chronic TMD were strikingly similar to associations observed in the US population. Assessments of representativeness in this demographically diverse group of community volunteers suggest that OPPERA case-control findings have good internal validity. Demographic associations with TMD were consistent with population benchmarks and with other studies, suggesting broad applicability of these OPPERA findings. Greater occurrence of TMD in non-Hispanic-Whites than in other racial/ethnic groups and the lack of a socioeconomic gradient contradicts the disparities seen in many other health conditions.


Early menopause is associated with lack of response to antiviral therapy in women with chronic hepatitis C.
Chronic hepatitis C (CHC) and liver fibrosis progress more rapidly in men and menopausal women than in women of reproductive age. We investigated the associations among menopause, sustained virologic response (SVR), and liver damage in patients with CHC. We performed a prospective study of 1000 consecutive, treatment-naïve patients 18 years of age and older with compensated liver disease from CHC. Liver biopsy samples were analyzed (for fibrosis, inflammation, and steatosis) before patients received standard antiviral therapy. From women (n = 442), we collected data on the presence, type, and timing of menopause; associated hormone and metabolic features; serum levels of interleukin-6; and hepatic tumor necrosis factor (TNF)-α. Postmenopausal women achieved SVRs less frequently than women of reproductive age (46.0% vs 67.5%; P < .0001) but as frequently as men (51.1%; P = .283). By multivariate regression analysis, independent significant predictors for women to not achieve an SVR were early menopause (odds ratio [OR], 8.055; 95% confidence interval [CI], 1.834-25.350), levels of γ-glutamyl transpeptidase (OR, 2.165; 95% CI, 1.364-3.436), infection with hepatitis C virus genotype 1 or 4 (OR, 3.861; 95% CI, 2.433-6.134), and cholesterol levels (OR, 0.985; 95% CI, 0.971-0.998). Early menopause was the only independent factor that predicted lack of an SVR among women with genotype 1 hepatitis C virus infection (OR, 3.933; 95% CI, 1.274-12.142). Baseline levels of liver inflammation, fibrosis, steatosis, serum interleukin-6 (P = .04), and hepatic TNF-α (P = .007) were significantly higher among postmenopausal women than women of reproductive age. Among women with CHC, early menopause was associated with a low likelihood of SVR, probably because of inflammatory factors that change at menopause.


The mechanism of binding of the second PDZ domain from the Protein Tyrosine Phosphatase-BL to the Adenomatous Polyposis Coli tumor suppressor.
Many biological processes are regulated by the interaction between protein domains and their corresponding binding partners. The PDZ domain is one of the most common protein-protein interaction modules in mammalian cells, whose role is to bind C-terminal sequences of specific targets. The second PDZ domain from the Protein Tyrosine Phosphatase-BL (PDZ2) binds to the C-terminal of Adenomatous Polyposis Coli protein (APC), one of the major tumor suppressor whose task is to regulate cell adhesion and proliferation. Here, we present a detailed kinetics analysis of the interaction between PDZ2 domain and a peptide mimicking the PDZ binding motif of APC. By analyzing data obtained at different experimental conditions, we propose a plausible mechanism for binding. Furthermore, a comparison between the dissociation rate constant measured by different methodologies allow us to identify an additional kinetic step, which is likely to arise from a conformational change of PDZ2 occurring after binding. The data are discussed on the light of previous work on PDZ domains.


Colorectal Cancer Develops Inherent Radiosensitivity That Can Be Predicted Using Patient-Derived Organoids.
Identifying colorectal cancer patient populations responsive to chemotherapy or chemoradiation therapy before surgery remains a challenge. Recently validated mouse protocols for organoid irradiation employ the single hit multi-target (SHMT) algorithm, which yields a single value, the D0, as a measure of inherent tissue radiosensitivity. Here, we translate these protocols to human tissue to evaluate radioresponsiveness of patient-derived organoids (PDO) generated from normal human intestines and rectal tumors of patients undergoing neoadjuvant therapy. While PDOs from adenomas with a logarithmically expanded Lgr5+ intestinal stem cell population retain the radioresistant phenotype of normal colorectal PDOs, malignant transformation yields PDOs from a large patient subpopulation displaying marked radiosensitivity due to reduced homologous recombination-mediated DNA repair. A proof-of-principle pilot clinical trial demonstrated that rectal cancer patient responses to neoadjuvant chemoradiation, including complete response, correlate closely with their PDO D0 values. Overall, upon transformation to colorectal adenocarcinoma, broad radiation sensitivity occurs in a large subset of patients that can be identified using SHMT analysis of PDO radiation responses. Analysis of inherent tissue radiosensitivity of patient-derived organoids may provide a readout predictive of neoadjuvant therapy response to radiation in rectal cancer, potentially allowing pretreatment stratification of patients likely to benefit from this approach.


Why actions for early treatment of febrile illnesses in children are delayed by caregivers.
The study explored why actions for early treatment of febrile illnesses in children are delayed by caregivers of children less than five years in five Local Government Areas (LGAs) of southwestern Nigeria using four indicators: caregivers' perception of illness, notion of causation and seriousness, belief in efficacy of selected pathway, and the decision making process. Seven types of febrile illnesses (yellow fever, typhoid fever, ordinary fever/malaria, hot body fever, rain fever, cold fever, and headache fever) were identified and yellow fever was perceived as the most severe type (60.8%). Only 24.2% correctly identified an infected mosquito bite as the cause of ordinary fever/malaria. Use of leftover drugs at home (55.6%) was the main action taken and fathers were the main decisionmakers in all cases. Empowerment of women for improved knowledge on causation, severity, and importance of prompt action for a healthy outcome is recommended.


Microneedles permit transdermal delivery of a skin-impermeant medication to humans.
Drugs with poor oral bioavailability usually are administered by hypodermic injection, which causes pain, poor patient compliance, the need for trained personnel, and risk of infectious disease transmission. Transdermal (TD) delivery provides an excellent alternative, but the barrier of skin's outer stratum corneum (SC) prevents delivery of most drugs. Micrometer-scale microneedles (MNs) have been used to pierce animal and human cadaver skin and thereby enable TD delivery of small molecules, proteins, DNA, and vaccines for systemic action. Here, we present a clinical study of MN-enhanced delivery of a medication to humans. Naltrexone (NTX) is a potent mu-opioid receptor antagonist used to treat opiate and alcohol dependence. This hydrophilic and skin-impermeant molecule was delivered from a TD patch to healthy human subjects with and without pretreatment of the skin with MNs. Whereas delivery from a standard NTX TD patch over a 72-h period yielded undetectable drug plasma levels, pretreatment of skin with MNs achieved steady-state plasma concentrations within 2 h of patch application and were maintained for at least 48 h. The MNs and NTX patch were well tolerated with mild systemic and application site side effects. The MN arrays were painless upon administration and not damaged during skin insertion, and no MNs were broken off into the skin. This human proof-of-concept study demonstrates systemic administration of a hydrophilic medication by MN-enhanced TD delivery. These findings set the stage for future human studies of skin-impermeant medications and biopharmaceuticals for clinical applications.


Medical problems of amputees.
The initial trauma of a major amputation and the medical disorders secondary to it are being studied at the Biomechanics Laboratory. Problems that arise from amputation and from the replacement of the limb by a prosthetic device are complex but interrelated, and approaches to study on basic, applied and clinical levels must be integrated. Three studies carried on at the Biomechanics Laboratory are described in this paper. In the field of amputation operations, it is believed that certain osteoplastic and myoplastic techniques, along with protection for nerve stumps and measures to provide optimum circulatory conditions, may restore biological and mechanical function to a greater degree than is possible with present procedures. From studies of the expenditure of energy during locomotion with and without assistive devices and during therapeutic exercise, it was found that crutch-walking is metabolically much more costly in energy for the amputee than walking with the suction-socket prosthesis; through these studies, it will be possible to define criteria for maximum work loads for disabled persons. In dermatological studies, although there are still unsolved problems, successful methods of treatment have been developed for certain bacterial and fungus infections, contact dermatitides and disorders resulting from edema.


Fluorescent spot test for detecting carbohydrate-splitting enzymes in meconium.
I describe a fluorescent spot test for detecting the acidic glycosidases in meconium. Meconium is mixed with various 4-methylumbelliferylglycosides at the appropriate pH. In the presence of carbohydrate-splitting enzymes, 4-methylumbelliferone, a strongly fluorescent compound, is released. The reaction mixture, when spotted on chromatography paper and viewed under ultraviolet light, exhibits a bright fluorescence for specimens containing acidic glycosidases. The technique is applicable to any test that involves production of fluorescent products from non-fluorescent reactants. The procedure was applied to meconium specimens selected because of increased albumin content known to be associated with cystic fibrosis in newborns. The presence of beta-D-fucosidase in meconium was correlated to increased albumin content. Detection of beta-D-fucosidase by the present procedure may be useful as a screening test for cystic fibrosis.


Impact of poster presentations on academic knowledge transfer from the oncologist perspective in Turkey.
Currently poster presentations offer a common visual medium for knowledge transfer by a wide range of health professionals. Our study aimed to determine the scientific importance of poster presentations for Medical and Radiation Oncologists. A survey form including 40 questions was distributed to a total of 131 oncologists experienced in poster presentations. One hundred completed survey forms were included in the study. Descriptive statistics and modified thematic analyses were performed on the responses. Overall 64% of the participants agreed that posters were a good medium for knowledge transfer. Some 88% agreed that concise and clear styled presentations would increase appealing interests for poster contents. Visual appearance was cited more influential than content of the subject; 70% of participants agreed that appearances of posters could help to draw more viewer attention. Of respondents, 63% believed that posters accompanied by their author were more attractive for congress attendees, and 33% of them declared that the halo effect of the poster presenter was also important. The present study indicated that intelligibility, appearance and visuality of posters are most important factors from the aspect of oncologist participants. Presenters must take into account these important points when preparing their academic posters.


Isolation and partial characterization of anglefish proglucagon.
Evidence is presented that proglucagon from anglefish islets is a single chain polypeptide with 78 amino acid residues and that the glucagon portion of it is liberated after tryptic cleavage. The most striking characteristic in the conversion of the anglerfish proglucagon to glucagon is that the cleaved peptide bonds display enormous sensitivity toward trypsin. Thus, conversion of the prohormone to glucagon occurs very rapidly within 3-10 min with a 1:500-1:1000 molar ratio of enzyme to substrate. Further, trypic cleavage of the anglerfish glucagon requires higher concentrations of trypsin (molar ratio 1:25 enzyme to substrate) and longer incubation time. The behavior of proglucagon and glucagon toward trypsin shows striking similarities with the tryptic conversion of anglerfish proinsulin to insulin.


Bilateral avascular necrosis of the femoral head following asynchronous postictal femoral neck fractures: a case report and review of the literature.
Bilateral avascular necrosis (AVN) following postictal bilateral fracture neck of the femur is a rare occurrence. Here, we report a case of bilateral AVN of the femoral head following an asynchronous bilateral postictal fracture neck of the femur. A 16-year-old autistic boy presented with left hip pain following an episode of seizures and radiographs showed Delbet type II fracture neck of the left femur. This was treated by closed reduction and cancellous screw fixation and skeletal traction for 6 weeks. At 3 months, follow-up radiograph showed union of the fracture, but he had developed segmental AVN with collapse of the head. At 8 months, the patient presented with pain in the right hip following another episode of seizures and radiograph of the pelvis showed a fresh Delbet type II fracture neck of the right femur with established AVN of the left femoral head. He underwent closed reduction and cancellous screw fixation of the right hip and implant exit of the left hip. At the 6-month follow-up after this surgery, his radiograph of the pelvis showed AVN with collapse and extrusion of the femoral head on the right side as well. Literature review shows an increased risk of fracture neck of the femur among epileptics. The incidence of AVN is maximum in Delbet type I, followed by Delbet type II and type III in that order. Although there are no clear guidelines on the management of post-traumatic AVN of the femoral head, the majority have reported that most of them will eventually develop arthritis and will require total hip replacement at a later date. Upon extensive literature search, no case report of bilateral fracture neck of the femur with bilateral AVN was found and hence this case was reported.


Altered spectra of hypermutation in DNA repair-deficient mice.
Affinity maturation of the humoral immune response is based on the ability of immunoglobulin variable genes to undergo a process of rapid and extensive somatic mutation followed by antigenic selection for antibodies with higher affinity. While the behaviour of this somatic hypermutation phenomenon has been well characterized over the last 20 years, the molecular mechanism responsible for inserting mutations has remained shrouded. To better understand this mechanism, we studied the interplay between hypermutation and other DNA associated activities such as DNA repair. There was no effect on the frequency and pattern of hypermutation in mice deficient for nucleotide excision repair, base excision repair and ataxia-telangiectasia mutated gene repair of double strand breaks. However, variable genes from mice lacking some components of mismatch repair had an increased frequency of tandem mutations and had more mutations of G and C nucleotides. These results suggest that the DNA polymerase(s) involved in the hypermutation pathway produces a unique spectra of mutations, which is then altered by mismatch repair and antigenic selection. We, also describe the differential pattern of expression of some nuclear DNA polymerases in hypermutating versus non-hypermutating B lymphocytes. The rapidly dividing germinal centre B cells expressed DNA polymerases alpha, beta, delta, epsilon and zeta, whereas the resting non-germinal centre cells did not express polymerases alpha or epsilon at detectable levels, although they did express polymerases beta, delta and zeta. The lack of expression of polymerase epsilon in the non-germinal centre cells suggests that this enzyme has a critical role in chromosomal replication but does not participate in DNA repair in these cells.


Humanized gene replacement in mice reveals the contribution of cancer stroma-derived HB-EGF to tumor growth.
Tumor progression is a complex process that involves the interaction of cancer cells with the cancer-surrounding stromal cells. The cancer stroma influences the cancer cell growth and metastatic potential. The EGF family growth factor HB-EGF is synthesized in cancer cells and plays pivotal roles in oncogenic transformation and tumor progression, but the contribution of HB-EGF expressed in tumor stromal cells to tumor growth remains unclear. In the present study, we found that HB-EGF was expressed in host-derived cancer stromal cells in xenograft and allograft mouse tumor models. CRM197 is a specific inhibitor of human HB-EGF that has no effect on mouse HB-EGF. To elucidate whether host-derived stromal HB-EGF contributes to tumor growth, we generated knock-in mice expressing a CRM197-inhibitable humanized mutant form of HB-EGF. Administration of CRM197 to humanized knock-in mice that were bearing tumors derived from human or mouse cancer cells revealed that inhibition of host-derived stromal HB-EGF by CRM197 significantly reduced tumor growth. These results suggest that HB-EGF in the cancer-associated stroma plays a significant role for tumor growth, and that the HB-EGF derived from the stroma, as well as that expressed by cancer cells, is a potential target for cancer therapy. The present results also suggest that the humanized HB-EGF knock-in mice could be utilized for pathophysiological studies of HB-EGF as well as the development of therapeutic strategies targeting HB-EGF.


Reduction of Ischemic Brain Edema by Combined use of Paeoniflorin and Astragaloside IV via Down-Regulating Connexin 43.
Paeoniflorin (PF) and astragaloside IV (AS-IV) have protective effects on cerebral ischemia. We aimed to test the effects of combined use of PF and AS-IV on ischemic brain edema and investigate whether the effects were dependent on connexin43 (Cx43). We detected the expression of Cx43 induced by PF and AS-IV after cerebral ischemia. We also examined the effects of combined use of PF and AS-IV on ischemic edema and further investigated the related pathways. We demonstrated PF and AS-IV decreased Cx43 and aquaporin4 (AQP4) associating with reduction of brain edema by dry-wet weight and brain-specific gravity methods after cerebral ischemia. Administration of PF and AS-IV displayed a further attenuation of brain edema with lower Cx43 levels. Meanwhile, Cx43 blockade inhibited AQP4 down-regulation by the two drugs. Moreover, phosphorylation of C-Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) were increased by PF and AS-IV, respectively. The effects of PF and AS-IV to down-regulate Cx43 were suppressed by JNK and ERK inhibitors, respectively. Our data indicate that PF and AS-IV alleviate ischemic brain edema, which has close relation to Cx43 down-regulation causing decrease of AQP4 via JNK and ERK pathways activation, respectively. Combined administration elicits synergistic effects on brain edema reduction. Copyright © 2017 John Wiley & Sons, Ltd.


Comparative assessment of articular cartilage and synovial membrane in experimental haemarthrosis.
The changes in articular cartilage and synovial membrane of the knee joints were studied in two groups of rabbits and Wistar rats with experimental haemarthrosis, electron microscopically. Hamarthrosis was produced in group 1 by a single autologous blood injection, in group 2 by intraarticular fracture of the femoral condyles. Samples were taken from the intact articular cartilage, the menisci and the infrapatellar portion of the synovial membrane 12 h to 20 days after intervention. Blood resorption occurs only in the synovial membrane. Fragmentation of erythrocytes and erythrocytophagy by synovial macrophages is documented. The different stages of intracellular digestion of erythrocyte fragments are traced down. Synovial fibroblasts do not participate in erythrocytophagy, although they disclose morphological signs of enhanced functional activity. The findings show changes in the matrix and chondrocytes within the articular cartilage and menisci, and presence of free erythrocytes and lipoprotein complexes amidst the collagen fibres of the matrix. The chondrocytes are poor in cell organelles, while the intracytoplasmic filaments, lipid droplets and glycogen granules are augmented in number. There is no evidence of erythrocytophagy by cartilage cells. On single blood injection in the joint, the ensuing changes are reversible, and the normal synovial membrane structure is restored much quicker than the articular cartilage.


Point-of-care screening to identify cognitive impairment in older adults.
To explore associations between results of a rapid screening tool for cognitive impairment and individual patient characteristics in a sample of patients receiving outpatient anticoagulation therapy who were not previously diagnosed with a dementia. Descriptive, nonexperimental, cross-sectional study. Pharmacist-managed anticoagulation clinic in Spokane, WA, from June 2006 to March 2007. 300 community-dwelling patients aged 60 years or older who had at least 6 months of outpatient anticoagulation therapy services. Following informed consent, demographic, medical history, medication history, anticoagulation therapy, and cognitive screening data were recorded from participant medical records, and a participant interview was performed using a standardized questionnaire and data collection form. Cognitive screening status (suggests cognitive impairment versus suggests dementia less likely) was used as an independent variable by which to compare patient demographics, medical history, medication history, and percent of out-of-range International Normalized Ratio (INR) visits. 55 of 300 participants (18.3%) with no previous diagnosis of a cognitive impairment were classified as "suggests cognitive impairment" based on the screening test. Presence or absence of cognitive impairment differed in those needing assistance with taking medications but was not associated with other sample characteristics, including percentage of visits with out-of-range INR value, gender, in-home care needs, age, and number of medical conditions. Screening at a convenient health care access point may lead to increased identification of community-dwelling elderly patients with unrecognized and undiagnosed cognitive impairment. Pharmacists are particularly well suited to conduct this screening because of the extended and frequent contact they have with patients in settings such as anticoagulation therapy clinics.


Carriage of extended-spectrum beta-lactamase-producing enterobacteriacae among internal medicine patients in Switzerland.
The incidence of extended-spectrum beta-lactamase producing-enterobacteriacae (ESBL-E) infection is rising worldwide. We aimed to determine the prevalence and nosocomial acquisition rate of ESBL-E as well as the risk factors for ESBL-E carriage and acquisition amongst patients consecutively admitted to 13 internal medicine units at our hospital who were not previously known to be ESBL-E carriers. We screened all patients admitted or transferred to internal medicine units for ESBL-E on admission and discharge using rectal swabs. Of 1072 patients screened, 51 (4.8%) were carriers of an ESBL-E at admission. Of 473 patients who underwent admission and discharge screening, 21 (4.4%) acquired an ESBL-E. On multivariate analysis, diabetes mellitus without end-organ complications (OR 2.87 [1.09-7.08]), connective tissue disease (OR 7.22 [1.17-44.59]), and liver failure (OR 8.39 [1.55-45.45]) were independent risk factors for carriage of an ESBL-E upon admission to hospital (area under the ROC curve, 0.68). Receipt of a first- or second-generation cephalosporin (OR 9.25 [2.22-37.82]), intra-hospital transfer (OR 6.68 [1.71-26.06]), and a hospital stay >21 days (OR 25.17 [4.18-151.68]) were associated with acquisition of an ESBL-E during hospitalisation; whilst admission from home was protective (OR 0.16 [0.06-0.39]) on univariate regression. No risk profile with sufficient accuracy to predict previously unknown carriage on admission or acquisition of ESBL-E could be developed using readily available patient information. ESBL-E carriage is endemic amongst internal medicine patients at our institution. We were unable to develop a clinical risk profile to accurately predict ESBL-E carriage amongst these patients.


Availability of Nanopore sequences in the genome taxonomy for Vibrionaceae systematics: Rumoiensis clade species as a test case.
Whole genome sequence comparisons have become essential for establishing a robust scheme in bacterial taxonomy. To generalize this genome-based taxonomy, fast, reliable, and cost-effective genome sequencing methodologies are required. MinION, the palm-sized sequencer from Oxford Nanopore Technologies, enables rapid sequencing of bacterial genomes using minimal laboratory resources. Here we tested the ability of Nanopore sequences for the genome-based taxonomy of Vibrionaceae and compared Nanopore-only assemblies to complete genomes of five Rumoiensis clade species: Vibrio aphrogenes, V. algivorus, V. casei, V. litoralis, and V. rumoiensis. Comparison of overall genome relatedness indices (OGRI) and multilocus sequence analysis (MLSA) based on Nanopore-only assembly and Illumina or hybrid assemblies revealed that errors in Nanopore-only assembly do not influence average nucleotide identity (ANI), in silico DNA-DNA hybridization (DDH), G+C content, or MLSA tree topology in Vibrionaceae. Our results show that the genome sequences from Nanopore-based approach can be used for rapid species identification based on the OGRI and MLSA.


An overview of recent advances and applications of FT-IR spectroscopy for quality, authenticity, and adulteration detection in edible oils.
Authenticity and adulteration detection are primary concerns of various stakeholders, such as researchers, consumers, manufacturers, traders, and regulatory agencies. Traditional approaches for authenticity and adulteration detection in edible oils are time-consuming, complicated, laborious, and expensive; they require technical skills when interpreting the data. Over the last several years, much effort has been spent in academia and industry on developing vibrational spectroscopic techniques for quality, authenticity, and adulteration detection in edible oils. Among them, Fourier transforms infrared (FT-IR) spectroscopy has gained enormous attention as a green analytical technique for the rapid monitoring quality of edible oils at all stages of production and for detecting and quantifying adulteration and authenticity in edible oils. The technique has several benefits such as rapid, precise, inexpensive, and multi-analytical; hence, several parameters can be predicted simultaneously from the same spectrum. Associated with chemometrics, the technique has been successfully implemented for the rapid detection of adulteration and authenticity in edible oils. After presenting the fundamentals, the latest research outcomes in the last 10 years on quality, authenticity, and adulteration detection in edible oils using FT-IR spectroscopy will be highlighted and described in this review. Additionally, opportunities, challenges, and future trends of FT-IR spectroscopy will also be discussed.


[On the specifics of the treatment effectiveness of the respiratory regulator in patients with bronchial asthma].
The authors show that the response of the respiratory system on additional resistance induced by RR depends on personal anxiety of the patient. Respiratory training with RR improves the condition of the respiratory system and psychological status of asthmatics. The training should be adjusted to psychological peculiarities of each patient and conducted according to individual program. Previous test for reaction of the respiratory system to additional exogenic resistance is needed.


Zebrafish hox paralogue group 2 genes function redundantly as selector genes to pattern the second pharyngeal arch.
The pharyngeal arches are one of the defining features of the vertebrates, with the first arch forming the mandibles of the jaw and the second forming jaw support structures. The cartilaginous elements of each arch are formed from separate migratory neural crest cell streams, which derive from the dorsal aspect of the neural tube. The second and more posterior crest streams are characterized by specific Hox gene expression. The zebrafish has a larger overall number of Hox genes than the tetrapod vertebrates, as the result of a duplication event in its lineage. However, in both zebrafish and mouse, there are just two members of Hox paralogue group 2 (PG2): Hoxa2 and Hoxb2. Here, we show that morpholino-mediated "knock-down" of both zebrafish Hox PG2 genes results in major defects in second pharyngeal arch cartilages, involving replacement of ventral elements with a mirror-image duplication of first arch structures, and accompanying changes to pharyngeal musculature. In the mouse, null mutants of Hoxa2 have revealed that this single Hox gene is required for normal second arch patterning. By contrast, loss-of-function of either zebrafish Hox PG2 gene individually has no phenotypic consequence, showing that these two genes function redundantly to confer proper pattern to the second pharyngeal arch. We have also used hoxb1a mis-expression to induce localized ectopic expression of zebrafish Hox PG2 genes in the first arch; using this strategy, we find that ectopic expression of either Hox PG2 gene can confer second arch identity onto first arch structures, suggesting that the zebrafish Hox PG2 genes act as "selector genes."