Disease-related pregnancy concerns and reproductive planning in women with inflammatory bowel diseases.
Women with inflammatory bowel diseases (IBD) endorse disease-related pregnancy concerns that influence parity. Improvements in IBD management have potentially altered reproductive planning. Additionally, the proportion of American women who choose not to have children is increasing. To explore the effect of disease-related pregnancy concerns on parity and reproductive planning in a subset of women with IBD. Cross-sectional qualitative phone survey in an academic gastroenterology practice. Questions included demographics, medical and reproductive history, future pregnancy plans, and if IBD affected pregnancy decision-making. Qualitative data were coded and frequencies and proportions calculated. The 129 female participants (31% response rate) were predominately white (85%), had at least some college education (97%) and a mean age of 34.3 years [standard deviation 6.2]. Some 60% had Crohn's disease and 30% had undergone IBD-related surgery. Half were nulliparae, 53% reported IBD-related pregnancy concerns and 57% desired future pregnancy. Women who desired a future pregnancy and had IBD-related concerns had higher parity than those without concerns (p=0.02). Women desiring a future pregnancy and those with Crohn's disease had increased IBD-related concerns. Only four (3.1%) women identified IBD-related concerns that led to a smaller family size than desired. IBD-related concerns appear to be less likely to affect a woman's planned family size than previously reported. Concern about adverse pregnancy outcomes is more common in women with Crohn's disease and those desiring future pregnancy, suggesting a need for targeted counselling to moderate risk perception.


Fluorescence-based system for measurement of electrophysiological changes in stretched cultured cardiomyocytes.
Acute or sustained stretch of cardiac tissue is known to play a key role in arrhythmogenesis. Using a fluorescence approach, we designed a system measuring calcium transients and transmembrane potential changes in monolayers of cultured cardiomyocytes under uniaxial elongation and electrical stimulation. Cardiac myocytes are seeded on a rectangular PDMS template held and stretched by a motorized linear guide system. Electrical stimulation is performed with two parallel carbon electrodes supplied by amplified pulses from a digital-to-analog converter. The cells are stained with either voltage- or calcium-sensitive dye (di-4-ANEPPS and Fluo-4 AM respectively). The two available excitation light sources are both current-controlled LED arrays (λ = 523 ± 45 nm for di-4-ANEPPS and λ = 505 ± 15 nm for Fluo-4 AM). The filtered emitted fluorescence (λ > 610 nm for di-4-ANEPPS and λ = 535 ± 25 nm for Fluo-4 AM) is transduced to current with a photodiode, converted to amplified voltage signals and digitized. The design and preliminary validation results are presented.


An N-terminal fragment of the gene 4 helicase/primase of bacteriophage T7 retains primase activity in the absence of helicase activity.
Primase and helicase activities of bacteriophage T7 are present in a single polypeptide coded by gene 4. Because the amino terminal region of the gene 4 protein contributes to primase activity, we constructed a truncated gene 4 encoding the N-terminal 271-aa residues. The truncated protein, purified from cells overexpressing the protein, is a dimer in solution; the full-length protein is a hexamer. Although the fragment is devoid of dTTPase and helicase activities, it catalyzes template-directed synthesis of di-, tri-, and tetranucleotides. The rates for tetraribonucleotide synthesis and for dinucleotide extension on a 20-nucleotide template are similar for the full-length and truncated proteins. However, the activity of the primase fragment is unaffected by dTTP whereas the primase activity of the full-length protein is stimulated >14-fold. The primase fragment is defective in the interaction with T7 DNA polymerase in that primer synthesis cannot be coupled to DNA synthesis.


[Randomized comparison of intravenous immunoglobulin and methylprednisolone pulse therapy in children with newly diagnosed idiopathic thrombocytic purpura. The Danish ITP Study Group].
Forty three children with newly diagnosed idiopathic thrombocytopenic purpura (ITP), platelet count (pl.c.) below 20 x 10(9)/l, and either clinically significant bleeding or failure to show a spontaneous platelet rise within three days of admission were randomly allocated to treatment with intravenous infusions of either immunoglobulin (IVIG) 1 g/kg or methylprednisolone (MPPT) 30 mg/kg on two consecutive days. Prompt induction of partial remission with pl.c. > 50 x 10(9)/l after 72 hours was seen in 21/23 given IVIG versus 10/20 given MPPT (exact p = 0.003); mean pl.c.s after 72 hours were 188 versus 77 x 10(9)/l (2p < 0.001). Poor responders were then given the alternative infusions in addition. After six days, complete remission with pl.c. > 150 x 10(9)/l was achieved in 16/23 versus 10/20 (p = 0.16). During six months follow-up, there were no significant differences regarding relapse rates or chronic course. Eleven children with relapse were crossed over to the alternative treatment arm: the estimated treatment effect in pl.c. after 72 hours was 134 x 10(9)/l in favour of IVIG. These results indicate that IVIG infusions may be preferable to high-dose corticosteroids as initial treatment for children with ITP.


Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress.
We examined the mechanism regulating the cellular levels of PNKP, the major kinase/phosphatase involved in the repair of oxidative DNA damage, and find that it is controlled by ATM phosphorylation and ubiquitylation-dependent proteasomal degradation. We discovered that ATM-dependent phosphorylation of PNKP at serines 114 and 126 in response to oxidative DNA damage inhibits ubiquitylation-dependent proteasomal degradation of PNKP, and consequently increases PNKP stability that is required for DNA repair. We have also purified a novel Cul4A-DDB1 ubiquitin ligase complex responsible for PNKP ubiquitylation and identify serine-threonine kinase receptor associated protein (STRAP) as the adaptor protein that provides specificity of the complex to PNKP. Strap(-/-) mouse embryonic fibroblasts subsequently contain elevated cellular levels of PNKP, and show elevated resistance to oxidative DNA damage. These data demonstrate an important role for ATM and the Cul4A-DDB1-STRAP ubiquitin ligase in the regulation of the cellular levels of PNKP, and consequently in the repair of oxidative DNA damage.


Genetic Mapping and Evolutionary Analyses of the Black Grain Trait in Barley.
Barley occupies the widest ecological area among the major cereal crops, thereby generating a high potential for adaptive genetic diversity against various environmental factors. Colored barley such as black grain barley has been suggested to result from environmental adaptation to biotic and abiotic stresses. Using one double haploid population (433 lines), plus three F5 recombinant inbred line (RIL) populations (1,009 lines), the black lemma and pericarp (Blp) gene was mapped between two Insertion/deletion (Indel) markers MC_1570156 and MC_162350 with a physical distance of 0.807 Mb, containing 21 annotated genes in the mapped interval. Whole-genome re-sequencing was performed on two Tibetan wild barley lines (X1 and W1) with black grain phenotype. The probable candidate genes for Blp were discussed based on gene functional annotation and gene sequence variation analyses. Thirteen polymorphic Indel markers covering the target genetic region were used to analyze 178 barley accessions including 49 black husk entries. Genotype-based clustering analyses showed that the black landraces of different geographical background may have evolved from a single origin. Our study represents a significant improvement on the genetic mapping of Blp and would facilitate future study on the characterization of the genetic basis underlying this interesting agronomic trait.


Can Impedance-pH Testing on Medications Reliably Identify Patients with GERD as Defined by Pathologic Esophageal Acid Exposure off Medications?
Impedance-pH testing (MII-pH) while patients are on acid suppression medications is frequently used to evaluate persistent reflux symptoms. The aim of this study was to determine whether MII-pH on medications can reliably identify patients with gastroesophageal reflux disease (GERD) as defined by pathologic esophageal acid exposure off medications, and to determine if there is a threshold of total reflux episodes on medications where pH testing off medications may be unnecessary. A retrospective review identified all patients between 1/2010 and 4/2017 who underwent MII-pH testing on PPI medications and subsequently had pH testing off medications. GERD was defined on pH testing off medications by an abnormal DeMeester Score (DMS) and on MII-pH on medications by ≥ 48 total reflux episodes. Patients with an abnormal DMS by MII-pH on medications were excluded. There were 71 patients, (22 males; 49 females), with a median age of 52 years. Based on ≥ 48 total reflux episodes by MII-pH testing on medications, 42 patients (59%) had GERD. When tested off medications, an abnormal DMS was present in 44 patients (62%). Among those with GERD based on impedance testing on medications, 31% did not have GERD based on pH testing off medications. Further, in the 29 patients with ≤ 48 total reflux episodes on MII-pH (normal test), 15 patients (52%) had pathologic acid exposure off medications. When there were > 73 reflux events with MII-pH on medications, all 15 patients in our series had pathologic acid exposure on pH testing off medications. MII-pH testing on medications in patients with refractory GERD symptoms does not reliably correlate with a diagnosis of GERD as defined by pathologic esophageal acid exposure off medications. The commonly used abnormal MII-pH test value of ≥ 48 total reflux episodes is not validated and should not be used. However, in our series, patients with > 73 total reflux episodes had a high likelihood of having pathologic acid exposure off medications. Overall, the preferred strategy to evaluate patients with persistent GERD symptoms on acid suppression therapy should be pH testing off medications.


EndoS2 is a unique and conserved enzyme of serotype M49 group A Streptococcus that hydrolyses N-linked glycans on IgG and α1-acid glycoprotein.
Many bacteria have evolved ways to interact with glycosylation functions of the immune system of their hosts. Streptococcus pyogenes [GAS (group A Streptococcus)] secretes the enzyme EndoS that cleaves glycans on human IgG and impairs the effector functions of the antibody. The ndoS gene, encoding EndoS, has, until now, been thought to be conserved throughout the serotypes. However, in the present study, we identify EndoS2, an endoglycosidase in serotype M49 GAS strains. We characterized EndoS2 and the corresponding ndoS2 gene using sequencing, bioinformatics, phylogenetic analysis, recombinant expression and LC-MS analysis of glycosidic activity. This revealed that EndoS2 is present exclusively, and highly conserved, in serotype M49 of GAS and is only 37% identical with EndoS. EndoS2 showed endo-β-N-acetylglucosaminidase activity on all N-linked glycans of IgG and on biantennary and sialylated glycans of AGP (α1-acid glycoprotein). The enzyme was found to act only on native IgG and AGP and to be specific for free biantennary glycans with or without terminal sialylation. GAS M49 expression of EndoS2 was monitored in relation to carbohydrates present in the culture medium and was linked to the presence of sucrose. We conclude that EndoS2 is a unique endoglycosidase in serotype M49 and differs from EndoS of other GAS strains by targeting both IgG and AGP. EndoS2 expands the repertoire of GAS effectors that modify key glycosylated molecules of host defence.


Association between Slip Severity and Muscle Synergies of Slipping.
Falls impose significant negative impacts to the US population and economy. A significant number of falls may be prevented via appropriate slip-responses since a strong relation exists between slips and falls. More importantly, as severe slips are more prone to result in a fall, identifying severe slippers along with the responsible factors for their adverse motor control and severe slipping should be the highest priority in fall prevention process. Previous studies have suggested that muscle synergies may be building blocks of the central nervous system in controlling motor tasks. Muscle synergies observed during slipping ('post-slip-initiation synergies' or 'just briefly,' 'slipping muscle synergies'), may represent the fundamental blocks of the neural control during slipping. Hence, studying the differences in slipping muscle synergies of mild and severe slippers can potentially reveal the differences in their neural control and subsequently, indicate the responsible factors for the adverse post-slip response in severe slippers. Even though the slipping muscle synergies have been investigated before, it still remains unclear on how the slip severity is associated with the slipping muscle synergies. More importantly, muscle synergies can be interpreted not only as neural blocks but also as physical sub-tasks of the main motor task. Hence, studying the differences of slipping synergies of mild and severe slippers would reveal the discrepancies in sub-tasks of their post-slip response. These discrepancies help pinpoint the malfunctioning sub-function associated with inadequate motor response seen in severe slippers. Twenty healthy subjects were recruited and underwent an unexpected slip (to extract their slipping synergies). Subjects were classified into mild and severe slippers based on their Peak Heel Speed. An independent t-test revealed several significant inter-group differences for muscle synergies of mild and severe slippers indicating differences in their neural control of slipping. A forward dynamic simulation was utilized to reveal the functionality of each synergy. Decomposition of slipping into sub-tasks (synergies), and finding the malfunctioning sub-task in severe slippers is important as it results in a novel targeted motor-rehabilitation technique that only aims to re-establish the impaired sub-task responsible for the adverse motor-response in severe slippers.


Positive correlation between the use of complementary and alternative medicine and internal health locus of control.
Because many people with chronic medical conditions use complementary and alternative medicine (CAM), understanding their psychosocial characteristics may be useful for identifying effective interventions. The health locus of control (HLOC) measures the perception of controlling one's own health outcomes by various attributes. People with a high internal HLOC believe that the outcome of their own health seeking is related to their behavior or personal investment. Earlier evidence has shown that a higher internal HLOC is a predictive factor of positive treatment outcomes. This study measured the correlation between the degree of CAM use and the level of HLOC. An online cross-sectional survey was conducted via public bulletin boards and invitational e-mails. Data from 123 usable responses were analyzed for bivariate correlation between CAM use and HLOC. Subjective reports of various medical modalities were classified into six CAM domains and one conventional biomedicine domain. Subscales of HLOC included internal, chance, and powerful others. Chronic conditions, health status, and demographics were self-reported. Internal HLOC significantly correlated with CAM use (Spearman's rho, P < .004) but not with conventional medicine use (Spearman's rho, P > .130). Further analysis of this correlation for those people with chronic conditions could not identify a particular domain used more by people with a high internal HLOC (P > .187), but the lesser use of conventional medicine was significant (P < .031). Complementary and alternative medicine is either empowering or has empowered patients to use CAM. People who use CAM may have a better prognosis and better management of chronic conditions.


A role for brassinosteroids in light-dependent development of Arabidopsis.
Although steroid hormones are important for animal development, the physiological role of plant steroids is unknown. The Arabidopsis DET2 gene encodes a protein that shares significant sequence identity with mammalian steroid 5 alpha-reductases. A mutation of glutamate 204, which is absolutely required for the activity of human steroid reductase, abolishes the in vivo activity of DET2 and leads to defects in light-regulated development that can be ameliorated by application of a plant steroid, brassinolide. Thus, DET2 may encode a reductase in the brassinolide biosynthetic pathway, and brassinosteroids may constitute a distinct class of phytohormones with an important role in light-regulated development of higher plants.


Rickettsia felis in cat fleas, Ctenocephalides felis parasitizing opossums, San Bernardino County, California.
Los Angeles and Orange Counties are known endemic areas for murine typhus in California; however, no recent reports of flea-borne rickettsioses are known from adjacent San Bernardino County. Sixty-five opossums (Didelphis virginiana) were trapped in the suburban residential and industrial zones of the southwestern part of San Bernardino County in 2007. Sixty out of 65 opossums were infested with fleas, primarily cat fleas, Ctenocephalides felis (Bouché, 1835). The flea minimum infection rate with Rickettsia felis was 13.3% in pooled samples and the prevalence was 23.7% in single fleas, with two gltA genotypes detected. In spite of historic records of murine typhus in this area, no evidence for circulation of R. typhi in fleas was found during the present study. Factors contributing to the absence of R. typhi in these cat fleas in contrast to its presence in cat fleas from Orange and Los Angeles Counties are unknown and need to be investigated further in San Bernardino County.


Telework After COVID: A "Silver Lining" for Workers with Disabilities?
Purpose The COVID pandemic was a severe blow to all workers, but it may ultimately have a silver lining for some workers with disabilities if it makes work from home easier and more acceptable. In addition, the pandemic is shaking up traditional workplace structures and causing employers to rethink how essential tasks can be done, which may broaden their views of workplace accommodations. We assess the potential for the pandemic to improve employment opportunities for people with disabilities. Methods This article analyzes pre-COVID data on disability and home-based work from three representative data sources-the American Community Survey, American Time Use Survey, and Current Population Survey. We employ both cross-tabulations and regressions to predict work at home. Results We find that workers with disabilities are more likely than those without disabilities both to work primarily from home and to do any work at home. This is true for both employees and self-employed workers. Workers with disabilities face similar wage gaps in on-site and home-based work, indicating that while increased availability of home-based work may create more employment opportunities for workers with disabilities, it is unlikely to erase wage disparities. While workers with disabilities are currently more likely to be working primarily from home, only 34% are in occupations with high potential for home-based work, compared to 40% of workers without disabilities. Conclusions Workers with disabilities are currently more likely to work from home and many may benefit from expanded work-at-home opportunities, but the types of jobs they hold constrain this potential. Research is needed to see how home-based work evolves as the economy recovers from the pandemic over the next several years.


MicroRNA programs in normal and aberrant stem and progenitor cells.
Emerging evidence suggests that microRNAs (miRNAs), an abundant class of ∼22-nucleotide small regulatory RNAs, play key roles in controlling the post-transcriptional genetic programs in stem and progenitor cells. Here we systematically examined miRNA expression profiles in various adult tissue-specific stem cells and their differentiated counterparts. These analyses revealed miRNA programs that are common or unique to blood, muscle, and neural stem cell populations and miRNA signatures that mark the transitions from self-renewing and quiescent stem cells to proliferative and differentiating progenitor cells. Moreover, we identified a stem/progenitor transition miRNA (SPT-miRNA) signature that predicts the effects of genetic perturbations, such as loss of PTEN and the Rb family, AML1-ETO9a expression, and MLL-AF10 transformation, on self-renewal and proliferation potentials of mutant stem/progenitor cells. We showed that some of the SPT-miRNAs control the self-renewal of embryonic stem cells and the reconstitution potential of hematopoietic stem cells (HSCs). Finally, we demonstrated that SPT-miRNAs coordinately regulate genes that are known to play roles in controlling HSC self-renewal, such as Hoxb6 and Hoxa4. Together, these analyses reveal the miRNA programs that may control key processes in normal and aberrant stem and progenitor cells, setting the foundations for dissecting post-transcriptional regulatory networks in stem cells.


Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice.
Inflammatory processes and foam cell formation are key determinants in the initiation and progression of atherosclerosis. Electrophilic nitro-fatty acids, byproducts of nitric oxide- and nitrite-dependent redox reactions of unsaturated fatty acids, exhibit antiinflammatory signaling actions in inflammatory and vascular cell model systems. The in vivo action of nitro-fatty acids in chronic inflammatory processes such as atherosclerosis remains to be elucidated. Herein, we demonstrate that subcutaneously administered 9- and 10-nitro-octadecenoic acid (nitro-oleic acid) potently reduced atherosclerotic lesion formation in apolipoprotein E-deficient mice. Nitro-fatty acids did not modulate serum lipoprotein profiles. Immunostaining and gene expression analyses revealed that nitro-oleic acid attenuated lesion formation by suppressing tissue oxidant generation, inhibiting adhesion molecule expression, and decreasing vessel wall infiltration of inflammatory cells. In addition, nitro-oleic acid reduced foam cell formation by attenuating oxidized low-density lipoprotein-induced phosphorylation of signal transducer and activator of transcription-1, a transcription factor linked to foam cell formation in atherosclerotic plaques. Atherosclerotic lesions of nitro-oleic acid-treated animals also showed an increased content of collagen and alpha-smooth muscle actin, suggesting conferral of higher plaque stability. These results reveal the antiatherogenic actions of electrophilic nitro-fatty acids in a murine model of atherosclerosis.


Time-Course Gene Set Analysis for Longitudinal Gene Expression Data.
Gene set analysis methods, which consider predefined groups of genes in the analysis of genomic data, have been successfully applied for analyzing gene expression data in cross-sectional studies. The time-course gene set analysis (TcGSA) introduced here is an extension of gene set analysis to longitudinal data. The proposed method relies on random effects modeling with maximum likelihood estimates. It allows to use all available repeated measurements while dealing with unbalanced data due to missing at random (MAR) measurements. TcGSA is a hypothesis driven method that identifies a priori defined gene sets with significant expression variations over time, taking into account the potential heterogeneity of expression within gene sets. When biological conditions are compared, the method indicates if the time patterns of gene sets significantly differ according to these conditions. The interest of the method is illustrated by its application to two real life datasets: an HIV therapeutic vaccine trial (DALIA-1 trial), and data from a recent study on influenza and pneumococcal vaccines. In the DALIA-1 trial TcGSA revealed a significant change in gene expression over time within 69 gene sets during vaccination, while a standard univariate individual gene analysis corrected for multiple testing as well as a standard a Gene Set Enrichment Analysis (GSEA) for time series both failed to detect any significant pattern change over time. When applied to the second illustrative data set, TcGSA allowed the identification of 4 gene sets finally found to be linked with the influenza vaccine too although they were found to be associated to the pneumococcal vaccine only in previous analyses. In our simulation study TcGSA exhibits good statistical properties, and an increased power compared to other approaches for analyzing time-course expression patterns of gene sets. The method is made available for the community through an R package.


Ineffective vaccination against solid tumors can be enhanced by hematopoietic cell transplantation.
Vaccination with tumor Ags has not been an effective treatment for solid tumors. The goal of the current study was to determine whether a combination of vaccination and hematopoietic cell transplantation (HCT) can effectively treat primary, disseminated, or metastatic CT26 and MC38 murine colon tumors. Vaccination of tumor-bearing mice with irradiated tumor cells and CpG adjuvant failed to alter progressive tumor growth. However, mice bearing primary, disseminated lung, or metastatic liver tumors were uniformly cured after administration of total body irradiation, followed by the transplantation of hematopoietic progenitor cells and T cells from syngeneic, but not allogeneic vaccinated donors. Requirements for effective treatment of tumors included irradiation of hosts, vaccination of donors with both tumor cells and CpG, transfer of both CD4(+) and CD8(+) T cells along with progenitor cells, and ability of donor cells to produce IFN-gamma. Irradiation markedly increased the infiltration of donor T cells into the tumors, and the combined irradiation and HCT altered the balance of tumor-infiltrating cells to favor CD8(+) effector memory T cells as compared with CD4(+)CD25(+)FoxP3(+) T regulatory cells. The combination of vaccination and autologous hematopoietic cell transplantation was also effective in treating tumors. In conclusion, these findings show that otherwise ineffective vaccination to solid nonhematologic tumors can be dramatically enhanced by HCT.


Neisseria cinerea isolates can adhere to human epithelial cells by type IV pilus-independent mechanisms.
In pathogenic Neisseria species the type IV pili (Tfp) are of primary importance in host-pathogen interactions. Tfp mediate initial bacterial attachment to cell surfaces and formation of microcolonies via pilus-pilus interactions. Based on genome analysis, many non-pathogenic Neisseria species are predicted to express Tfp, but aside from studies on Neisseria elongata, relatively little is known about the formation and function of pili in these organisms. Here, we have analysed pilin expression and the role of Tfp in Neisseria cinerea. This non-pathogenic species shares a close taxonomic relationship to the pathogen Neisseria meningitidis and also colonizes the human oropharyngeal cavity. Through analysis of non-pathogenic Neisseria genomes we identified two genes with homology to pilE, which encodes the major pilin of N. meningitidis. We show which of the two genes is required for Tfp expression in N. cinerea and that Tfp in this species are required for DNA competence, similar to other Neisseria. However, in contrast to the meningococcus, deletion of the pilin gene did not impact the association of N. cinerea to human epithelial cells, demonstrating that N. cinerea isolates can adhere to human epithelial cells by Tfp-independent mechanisms.


Characterization of a germ-line proliferation mutation in C. elegans.
The C. elegans germ line is generated by extensive proliferation of the two germ-line progenitor cells present in newly hatched larvae. We describe genetic and phenotypic characterization of glp-4, a locus whose product is required for normal proliferation of the germ line. glp-4(bn2ts) mutant worms raised at the restrictive temperature contain approximately 12 germ nuclei, in contrast to the 700-1000 present in wild-type adults. The few germ cells present in sterile glp-4 adults appear to be arrested at prophase of the mitotic cell cycle. This cell-cycle disruption prevents the germ cells from entering meiosis and differentiating into gametes. Shifting sterile glp-4 worms to the permissive temperature enables their germ cells to undergo extensive proliferation and form gametes, demonstrating that the bn2-induced cell-cycle arrest is reversible and that proliferation and differentiation of germ cells can be uncoupled from development of the somatic gonad. The glp-4(bn2ts) mutation can be used to generate large populations of worms that are severely depleted in germ cells, facilitating determination of whether any gene of interest is expressed in the germ line or soma or both.


Population structure of Cercospora kikuchii, the causal agent of Cercospora leaf blight and purple seed stain in soybean.
Random amplified polymorphic DNA (RAPD) and microsatellite-primed polymerase chain reaction (MP-PCR) were used to characterize 164 isolates of Cercospora kikuchii, most of which were collected from Louisiana. Plant tissue (seeds versus leaves), but not host cultivar, had a significant impact on pathogen population differentiation. Cluster analysis showed that the Louisiana population was dominated by a primary lineage (group I) with only a few Louisiana isolates belonging to the minor lineage that also included the non-Louisiana isolates (group II). A previous study showed that isolates could be differentiated according to vegetative compatibility groups (VCGs). However, RAPD and MP-PCR data demonstrated that isolates of C. kikuchii were not generally clustered according to these VCGs. Furthermore, genetic relationships within and between VCGs were examined using sequences of the intergenic spacer region of rDNA. These analyses showed that VCG is not an indicator of evolutionary lineage in this fungus. Our results suggest the likely existence of a cryptically functioning sexual stage in some portion of the C. kikuchii population.


Catalytic versatility and backups in enzyme active sites: the case of serum paraoxonase 1.
The origins of enzyme specificity are well established. However, the molecular details underlying the ability of a single active site to promiscuously bind different substrates and catalyze different reactions remain largely unknown. To better understand the molecular basis of enzyme promiscuity, we studied the mammalian serum paraoxonase 1 (PON1) whose native substrates are lipophilic lactones. We describe the crystal structures of PON1 at a catalytically relevant pH and of its complex with a lactone analogue. The various PON1 structures and the analysis of active-site mutants guided the generation of docking models of the various substrates and their reaction intermediates. The models suggest that promiscuity is driven by coincidental overlaps between the reactive intermediate for the native lactonase reaction and the ground and/or intermediate states of the promiscuous reactions. This overlap is also enabled by different active-site conformations: the lactonase activity utilizes one active-site conformation whereas the promiscuous phosphotriesterase activity utilizes another. The hydrolysis of phosphotriesters, and of the aromatic lactone dihydrocoumarin, is also driven by an alternative catalytic mode that uses only a subset of the active-site residues utilized for lactone hydrolysis. Indeed, PON1's active site shows a remarkable level of networking and versatility whereby multiple residues share the same task and individual active-site residues perform multiple tasks (e.g., binding the catalytic calcium and activating the hydrolytic water). Overall, the coexistence of multiple conformations and alternative catalytic modes within the same active site underlines PON1's promiscuity and evolutionary potential.


T cell-mediated curation and restructuring of tumor tissue coordinates an effective immune response.
Antigen-specific T cells traffic to, are influenced by, and create unique cellular microenvironments. Here we characterize these microenvironments over time with multiplexed imaging in a melanoma model of adoptive T cell therapy and human patients with melanoma treated with checkpoint inhibitor therapy. Multicellular neighborhood analysis reveals dynamic immune cell infiltration and inflamed tumor cell neighborhoods associated with CD8+ T cells. T cell-focused analysis indicates T cells are found along a continuum of neighborhoods that reflect the progressive steps coordinating the anti-tumor immune response. More effective anti-tumor immune responses are characterized by inflamed tumor-T cell neighborhoods, flanked by dense immune infiltration neighborhoods. Conversely, ineffective T cell therapies express anti-inflammatory cytokines, resulting in regulatory neighborhoods, spatially disrupting productive T cell-immune and -tumor interactions. Our study provides in situ mechanistic insights into temporal tumor microenvironment changes, cell interactions critical for response, and spatial correlates of immunotherapy outcomes, informing cellular therapy evaluation and engineering.


Thalamo-Cortical Disruption Contributes to Short-Term Memory Deficits in Patients with Medial Temporal Lobe Damage.
Short-term (STM) and long-term memory (LTM) have largely been considered as separate brain systems reflecting fronto-parietal and medial temporal lobe (MTL) functions, respectively. This functional dichotomy has been called into question by evidence of deficits on aspects of working memory in patients with MTL damage, suggesting a potentially direct hippocampal contribution to STM. As the hippocampus has direct anatomical connections with the thalamus, we tested the hypothesis that damage to thalamic nuclei regulating cortico-cortical interactions may contribute to STM deficits in patients with hippocampal dysfunction. We used diffusion-weighted magnetic resonance imaging-based tractography to identify anatomical subdivisions in patients with MTL epilepsy. From these, we measured resting-state functional connectivity with detailed cortical divisions of the frontal, temporal, and parietal lobes. Whereas thalamo-temporal functional connectivity reflected LTM performance, thalamo-prefrontal functional connectivity specifically predicted STM performance. Notably, patients with hippocampal volume loss showed thalamic volume loss, most prominent in the pulvinar region, not detected in patients with normal hippocampal volumes. Aberrant thalamo-cortical connectivity in the epileptic hemisphere was mirrored in a loss of behavioral association with STM performance specifically in patients with hippocampal atrophy. These findings identify thalamo-cortical disruption as a potential mechanism contributing to STM deficits in the context of MTL damage.


Cognitive decline in the elderly after surgery and anaesthesia: results from the Oxford Project to Investigate Memory and Ageing (OPTIMA) cohort.
Concerns have been raised about the effects on cognition of anaesthesia for surgery, especially in elderly people. We recorded cognitive decline in a cohort of 394 people (198 women) with median (IQR) age at recruitment of 72.6 (66.6-77.8) years, of whom 109 had moderate or major surgery during a median (IQR) follow-up of 4.1 (2.0-7.6) years. Cognitive decline was more rapid in people who on recruitment were: older, p = 0.0003; male, p = 0.027; had worse cognition, p < 0.0001; or carried the ε4 allele of apoliprotein E (APOEε4), p = 0.008; and after an operation if cognitive impairment was already diagnosed, p = 0.0001. Cognitive decline appears to accelerate after surgery in elderly patients diagnosed with cognitive impairment, but not other elderly patients.


Circadian rhythms vary over the growing season and correlate with fitness components.
Circadian clocks have evolved independently in all three domains of life, suggesting that internal mechanisms of time-keeping are adaptive in contemporary populations. However, the performance consequences of either discrete or quantitative clock variation have rarely been tested in field settings. Clock sensitivity of diverse segregating lines to the environment remains uncharacterized as do the statistical genetic parameters that determine evolutionary potential. In field studies with Arabidopsis thaliana, we found that major perturbations to circadian cycle length (referred to as clock period) via mutation reduce both survival and fecundity. Subtler adjustments via genomic introgression of naturally occurring alleles indicated that clock periods slightly >24 hr were adaptive, consistent with prior models describing how well the timing of biological processes is adjusted within a diurnal cycle (referred to as phase). In segregating recombinant inbred lines (RILs), circadian phase varied up to 2 hr across months of the growing season, and both period and phase expressed significant genetic variances. Performance metrics including developmental rate, size and fruit set were described by principal components (PC) analyses and circadian parameters correlated with the first PC, such that period lengths slightly >24 hr were associated with improved performance in multiple RIL sets. These experiments translate functional analyses of clock behaviour performed in controlled settings to natural ones, demonstrating that quantitative variation in circadian phase is highly responsive to seasonally variable abiotic factors. The results expand upon prior studies in controlled settings, showing that discrete and quantitative variation in clock phenotypes correlates with performance in nature.


Mechanisms of probiotic action: Implications for therapeutic applications in inflammatory bowel diseases.
Probiotics are defined as nonpathogenic living microorganisms, including some commensal bacterial flora, which have beneficial effects on host health and disease prevention and/or treatment. Clinical trials have shown beneficial effects of probiotics on several human diseases, such as inflammatory bowel diseases (IBDs), which are among the most-studied diseases testing probiotics as a potential therapy. However, a significant question regarding clinical use of probiotics is the mechanism underlying the wide range of actions. Studies discussed in this review suggest 3 distinct cellular and molecular mechanisms for probiotic regulation in IBD therapy: 1) Probiotics block pathogenic bacterial effects by producing bactericidal substances and competing with pathogens and toxins for adherence to the intestinal epithelium; 2) Probiotics regulate immune responses by enhancing the innate immunity and modulating pathogen-induced inflammation via toll-like receptor-regulated signaling pathways; and 3) Probiotics regulate intestinal epithelial homeostasis by promoting intestinal epithelial cell survival, enhancing barrier function, and stimulating protective responses. Probiotics modulate host cell signaling pathways, including Akt, mitogen-activated protein kinases, and nuclear factor-kappaB to mediate these intestinal epithelial functions. It is hoped that developing a mechanistic understanding of probiotic action will provide the rationale to support the development of new hypothesis-driven studies to define the clinical efficacy in preventive, adjunctive, or alternative treatments for IBD.


The PERK Branch of the Unfolded Protein Response Safeguards Protein Homeostasis and Mesendoderm Specification of Human Pluripotent Stem Cells.
Cardiac development involves large-scale rearrangements of the proteome. How the developing cardiac cells maintain the integrity of the proteome during the rapid lineage transition remains unclear. Here it is shown that proteotoxic stress visualized by the misfolded and/or aggregated proteins appears during early cardiac differentiation of human pluripotent stem cells and is resolved by activation of the PERK branch of unfolded protein response (UPR). PERK depletion increases misfolded and/or aggregated protein accumulation, leading to pluripotency exit defect and impaired mesendoderm specification of human pluripotent stem cells. Mechanistically, it is found that PERK safeguards mesendoderm specification through its conserved downstream effector ATF4, which subsequently activates a novel transcriptional target WARS1, to cope with the differentiation-induced proteotoxic stress. The results indicate that protein quality control represents a previously unrecognized core component of the cardiogenic regulatory network. Broadly, these findings provide a framework for understanding how UPR is integrated into the developmental program by activating the PERK-ATF4-WARS1 axis.


Effects of room environment and nursing experience on clinical blood pressure measurement: an observational study.
This study aimed to examine the effects of measurement room environment and nursing experience on the accuracy of manual auscultatory blood pressure (BP) measurement. A training database with 32 Korotkoff sounds recordings from the British Hypertension Society was played randomly to 20 observers who were divided into four groups according to the years of their nursing experience (i.e. ≥10 years, 1-9 years, nursing students with frequent training, and those without any medical background; five observers in each group). All the observers were asked to determine manual auscultatory systolic blood pressure (SBP) and diastolic blood pressure (DBP) both in a quiet clinical assessment room and in a noisy nurse station area. This procedure was repeated on another day, yielding a total of four measurements from each observer (i.e. two room environments and two repeated determinations on 2 separate days) for each Korotkoff sound. The measurement error was then calculated against the reference answer, with the effects of room environment and nursing experience of the observer investigated. Our results showed that there was no statistically significant difference for BPs measured under both quiet and noisy environments (P>0.80 for both SBP and DBP). However, there was a significant effect on the measurement accuracy between the observer groups (P<0.001 for both SBP and DBP). The nursing students performed best with overall SBP and DBP errors of -0.8±2.4 and 0.1±1.8 mmHg, respectively. The SBP measurement error from the nursing students was significantly smaller than that for each of the other three groups (all P<0.001). Our results indicate that frequent nursing trainings are important for nurses to achieve accurate manual auscultatory BP measurement.


AMPK Contributes to Cardioprotective Effects of Pterostilbene Against Myocardial Ischemia- Reperfusion Injury in Diabetic Rats by Suppressing Cardiac Oxidative Stress and Apoptosis.
Pterostilbene (PT) exerts antidiabetic effects by decreasing blood glucose and modulating lipid metabolism and has been shown to attenuate myocardial ischemia-reperfusion (IR) injury in non-diabetic subjects. However, whether PT can protect against myocardial IR injury in diabetes is unknown. AMPK stimulation is indispensable in offering cardioprotection against myocardial IR injury in diabetes by limiting cardiac apoptosis. Thus, we hypothesized that PT may confer protection against myocardial IR injury in diabetes via AMPK activation. Sprague-Dawley rats at eight weeks of diabetes induction (induced by an intravenous dose of 65 mg/kg streptozotocin) were administered with vehicle or PT (20 and 40 mg/kg/day, p.o.) for four weeks (starting from week 9 to 12). At the end of week 12, myocardial IR injury was induced by subjecting the diabetic rats to 30 minutes of coronary artery ligation and followed by 2 hours of reperfusion. In in vitro studies, rat primary cardiomyocytes were incubated with low glucose (LG, 5.5 mM) or high glucose (HG, 30 mM) and exposed to 45 minutes hypoxia and 2 hours reoxygenation in the presence or absence of PT (0.5 µM) or the AMPK inhibitor compound C (CC, 5 µM). PT significantly reduced post-ischemic cardiac infarct size, oxidative stress, plasma lactate dehydrogenase (LDH), creatine kinase-MB levels and apoptosis in diabetic rats. In cardiomyocytes, PT decreased hypoxia/ reoxygenation-induced oxidative stress, attenuated LDH and cleaved caspase3/caspase3 ratio and increased Bcl-2/Bax ratio and AMPK phosphorylation. However, CC administration blunted the cardioprotective effects of PT both in vivo and in vitro. Suppressing cardiac oxidative stress and apoptosis via AMPK stimulation may represent a primary mechanism whereby pterostilbene attenuates diabetic myocardial IR injury.


High-resolution NMR correlation spectra of disordered solids.
We show how high-resolution NMR spectra can be obtained for solids for which the spectra are normally broadened due to structural disorder. The method relies on correlations in the chemical shifts between pairs of coupled spins. It is found experimentally that there are strong correlations in the chemical shifts between neighboring spins in both phosphorus-31 and carbon-13 spectra. These correlations can be exploited not only to provide resolution in two-dimensional spectra, but also to yield "chains" of correlated chemical shifts, constituting a valuable new source of structural information for disordered materials.


Organic-inorganic macroion coacervate complexation.
Coacervate complexes that are liquid-liquid separated complex materials are often formed by stoichiometrically mixing oppositely charged polyelectrolytes in salted aqueous solution. Entropy-driven ion pairing, resulting from the release of counterions near polyelectrolytes, has been identified as the primary driving force for coacervate formation between oppositely charged polyelectrolytes, including proteins and DNA, in aqueous solution. In this work we have examined the complexation between net neutral zwitterionic poly(sulfobetaine methacrylate) (PSBMA) and inorganic polyoxometalate (POM) polyanions in LiCl aqueous solutions. Biphasic liquid-like coacervate complexes can be formed over a much broader range of POM-to-PSBMA molar ratio and LiCl concentration than that for conventional polyelectrolyte coacervate complexation. Composition analysis of the dried supernatant and dense coacervate has confirmed that both PSBMA and POM macroions are primarily present in the dense coacervate as the macroion-rich phase in contrast to the presence of LiCl solely in the supernatant as the macroion-poor phase. The increase of net charge negativity of PSBMA and supernatant conductivity suggests stronger binding of PSBMA with POM anions than monovalent Cl-, resulting in the release of bound Cl- anions to the aqueous solution for the formation of PSBMA-POM coacervates in LiCl solution. All experimental evidence has demonstrated the generality of ion-pairing induced coacervate complexation with net neutral zwitterionic polymers and multivalent inorganic nanomaterials. The complexation between organic and inorganic macroions could give insights into many supramolecular assembly processes in nature and also lead to a new paradigm in developing hybrid macroionic materials for emerging applications from green catalysis to nanomedicine.


A theoretical study of lipid accumulation in the liver-implications for nonalcoholic fatty liver disease.
A hallmark of the nonalcoholic fatty liver disease is the accumulation of lipids. We developed a mathematical model of the hepatic lipid dynamics to simulate the fate of fatty acids in hepatocytes. Our model involves fatty acid uptake, lipid oxidation, and lipid export. It takes into account that storage of triacylglycerol within hepatocytes leads to cell enlargement reducing the sinusoids radius and impairing hepatic microcirculation. Thus oxygen supply is reduced, which impairs lipid oxidation. The analysis of our model revealed a bistable behavior (two stable steady states) of the system, in agreement with histological observations showing distinct areas of lipid accumulation in lobules. The first (healthy) state is characterized by intact lipid oxidation and a low amount of stored lipids. The second state in our model may correspond to the steatotic cell; it is marked by a high amount of stored lipids and a reduced lipid oxidation caused by impaired oxygen supply. Our model stresses the role of insufficient oxygen supply for the development of steatosis. We discuss implications of our results in regard to the experimental design aimed at exploring lipid metabolism reactions under steatotic conditions. Moreover, the model helps to understand the reversibility of lipid accumulation and predicts the reversible switch to show hysteresis. The system can switch from the steatotic state back to the healthy state by reduction of fatty acid uptake below the threshold at which steatosis started. The reversibility corresponds to the observation that caloric restriction can reduce the lipid content in the liver.


Institutional responsibilities and workload of faculty in baccalaureate dental hygiene programs.
This article reports the results of a study conducted to determine the institutional responsibilities and workload of full-time faculty in baccalaureate dental hygiene programs. A mail questionnaire was sent to program administrators, who were asked to distribute it to faculty. Faculty reported an average work week of 50.5 hours, which includes 46.9 hours spent on paid activities and 3.6 hours spent on unpaid activities. In specific workload activities, the majority of faculty time was spent on teaching undergraduate students (56.8 percent), institutional service (14.9 percent), and research/scholarship (9.5 percent). Forty-seven percent of the faculty described their primary professional research as program/curriculum design, and 78 percent were not engaged in funded research. The most common form of scholarship reported by faculty was presentations at professional meetings; this outnumbered all other types of scholarly activity, including publications. Faculty spent significantly more time than they preferred on teaching undergraduate students and institutional service. Faculty spent significantly less time than they preferred on teaching graduate/first professional students, research/scholarship, professional growth, and public service. We recommend that future studies compare workloads and scholarly production of dental hygiene faculty in associate and baccalaureate degree programs. We also recommend future longitudinal assessments of institutional responsibilities and workload of baccalaureate dental hygiene faculty.


Future research needs for evaluating the integration of mental health and substance abuse treatment with primary care.
Research needs are many in the current health care environment. In this article, we describe a novel method developed by the Agency for Healthcare Research and Quality (AHRQ) Evidence-based Practice Center Program for prioritizing areas for future research. Using a recent- ly published systematic review as a foundation, investigators worked with a diverse group of 10 stakeholders to identify and prioritize research needs. We enumerate 13 high-priority research needs, as determined by stakeholders who represented researchers, funders, health care providers, and patients and families, and discuss considerations for specific study designs. Our findings suggest that future research on integrating mental health and primary care should focus first on a) identifying methods of integrating primary care into specialty mental health settings, b) identifying cross-cutting strategies for integration across multiple mental health diagnostic categories as opposed to a separate strategy for each diagnostic category, and c) examining the use of information technology for integrating mental and general medical health care. Other priorities for consideration include examining the economic and organizational sus- tainability of successful integration models, identifying dissemination methods for various settings, examining the business case for integration as well as methods of payment, assessing the cost-effectiveness of integration, and identifying key components of successful strategies. The importance of sustainability and economic justification for integrated care strategies was a recurring theme in discussions with the stake- holders. The ability to sustain integrated care in everyday practice remains to be proved and will depend in part on the level of incentives and sup- port provided through payment system reform, as well as the ability of practices to provide care efficiently.


Comparison of multilocus sequence typing and pulsed-field gel electrophoresis as tools for typing Staphylococcus aureus isolates in a microepidemiological setting.
Multilocus sequence typing (MLST) of Staphylococcus aureus is well suited to the study of global or long-term epidemiology, but its role in local epidemiology has not been defined. The present study has compared MLST with pulsed-field gel electrophoresis (PFGE) by using S. aureus isolates associated with carriage and disease in a busy regional renal unit. One hundred forty-four patients were prospectively recruited, of whom 103 were receiving hemodialysis and 41 were on continuous ambulatory peritoneal dialysis. Three nasal swab specimens were obtained 1 month apart on entering the study. A nasal swab was positive for S. aureus on at least one occasion in 50 patients (35%). Typing of the 104 carriage isolates demonstrated 21 PFGE types and 21 sequence types (STs). Thirty-one carriers had two or more positive nasal swabs; of these, the isolates in all swabs from a given carrier had identical PFGE types for 29 carriers; the isolates in all of the same 29 swabs had identical STs. The carriage strain in two patients changed both PFGE type and STs during the period of swabbing. Eight patients (6%) had an episode of S. aureus bacteremia during the 12-month study period, and two of these were nasal carriers. One of these invasive isolates had the same PFGE type and ST as the carriage isolate. There were no differences between Simpson's index of diversity for PFGE and Simpson's index of diversity for MLST for both invasive and carriage isolates, suggesting that the two methods have very similar discriminatory abilities. We conclude that PFGE and MLST performed equally in this study.


The structure of state health agencies: a strategic analysis.
Leaders in public organizations are adopting many private sector management practices to control costs and increase efficiency. Nowhere is this more evident than among state health agencies. State health agencies were encouraged to change the way they operate by the 1988 Institute of Medicine (IOM) report on The Future of Public Health. This report portrayed public health as being in disarray. To address major deficiencies identified by the IOM study, some public health leaders have reevaluated their environments, reconfigured their organizations, and adopted a strategic mindset. The purpose of this research is to explore the various organizational configurations of state health agencies. Replicating methods used in studies of private sector organizations, five distinct strategic configurations or archetypes were identified. This comprehensive public health agency taxonomy will assist future researchers in analyzing public health organizations' environments, structures, and strategies.


A chromosome 3-encoded repressor of the human telomerase reverse transcriptase (hTERT) gene controls the state of hTERT chromatin.
Telomerase is crucial for human carcinogenesis. The limiting component of telomerase activity is telomerase reverse transcriptase (hTERT), undetectable in differentiated somatic cells but present in most tumor cells. There is evidence that hTERT transcription is shut down by a repressor in normal cells, but the mechanisms that turn on or maintain expression in tumor cells are not understood. To identify cis-acting regulatory elements, we scanned the hTERT gene for nuclease sensitive sites. In tumor cells and in in vitro transformed fibroblasts that contain hTERT mRNA, we detected a pattern of nuclease-sensitive sites in the second intron different from that in normal fibroblasts. To test whether the chromatin state characterized by the increased nuclease sensitivity plays a role in tumor-specific hTERT expression, we used a telomerase-positive breast carcinoma line, 21NT. Introduction of a normal chromosome 3 into these cells is known to down-regulate hTERT expression, probably through transcriptional silencing. 21NT cells displayed a similar pattern of micrococcal nuclease (MNase) sensitivity to other telomerase-positive lines, whereas the hTERT chromatin of the chromosome 3-hybrids resembled that of normal fibroblasts. In segregants that had lost the normal chromosome 3, the MNase sensitivity pattern characteristic of telomerase-positive cells was restored, and some (but not all) re-expressed the hTERT gene. The simplest model compatible with these results, and with data on the mapping of an hTERT repressor on chromosome 3, is that hTERT expression in tumor cells depends on an open state of intron 2-chromatin. We propose that, during the development of the breast carcinoma from which the 21NT cell line was derived, loss of function of this repressor led to chromatin remodeling necessary (but probably not sufficient) for expression of the hTERT gene. An improved understanding of the precise mechanism of hTERT dysregulation in human cancer may well find applications in the development of antitelomerase cancer therapy.


Twelve tips for efficient procedural simulation.
Procedural simulation (PS) is increasingly being used worldwide in healthcare for training caregivers in psychomotor competencies. It has been demonstrated to improve learners' confidence and competence in technical procedures, with consequent positive impacts on patient outcomes and safety. Several frameworks can guide healthcare educators in using PS as an educational tool. However, no theory-informed practical framework exists to guide them in including PS in their training programs. We present 12 practical tips for efficient PS training that translates educational concepts from theory to practice, based on the existing literature. In doing this, we aim to help healthcare educators to adequately incorporate and use PS both for optimal learning and for transfer into professional practice.


Influence of stochastic domain growth on pattern nucleation for diffusive systems with internal noise.
Numerous mathematical models exploring the emergence of complexity within developmental biology incorporate diffusion as the dominant mechanism of transport. However, self-organizing paradigms can exhibit the biologically undesirable property of extensive sensitivity, as illustrated by the behavior of the French-flag model in response to intrinsic noise and Turing's model when subjected to fluctuations in initial conditions. Domain growth is known to be a stabilizing factor for the latter, though the interaction of intrinsic noise and domain growth is underexplored, even in the simplest of biophysical settings. Previously, we developed analytical Fourier methods and a description of domain growth that allowed us to characterize the effects of deterministic domain growth on stochastically diffusing systems. In this paper we extend our analysis to encompass stochastically growing domains. This form of growth can be used only to link the meso- and macroscopic domains as the "box-splitting" form of growth on the microscopic scale has an ill-defined thermodynamic limit. The extension is achieved by allowing the simulated particles to undergo random walks on a discretized domain, while stochastically controlling the length of each discretized compartment. Due to the dependence of diffusion on the domain discretization, we find that the description of diffusion cannot be uniquely derived. We apply these analytical methods to two justified descriptions, where it is shown that, under certain conditions, diffusion is able to support a consistent inhomogeneous state that is far removed from the deterministic equilibrium, without additional kinetics. Finally, a logistically growing domain is considered. Not only does this show that we can deal with nonmonotonic descriptions of stochastic growth, but it is also seen that diffusion on a stationary domain produces different effects to diffusion on a domain that is stationary "on average."


Mitochondrial ATP synthase dimers spontaneously associate due to a long-range membrane-induced force
Adenosine triphosphate (ATP) synthases populate the inner membranes of mitochondria, where they produce the majority of the ATP required by the cell. From yeast to vertebrates, cryoelectron tomograms of these membranes have consistently revealed a very precise organization of these enzymes. Rather than being scattered throughout the membrane, the ATP synthases form dimers, and these dimers are organized into rows that extend for hundreds of nanometers. The rows are only observed in the membrane invaginations known as cristae, specifically along their sharply curved edges. Although the presence of these macromolecular structures has been irrefutably linked to the proper development of cristae morphology, it has been unclear what drives the formation of the rows and why they are specifically localized in the cristae. In this study, we present a quantitative molecular-simulation analysis that strongly suggests that the dimers of ATP synthases organize into rows spontaneously, driven by a long-range attractive force that arises from the relief of the overall elastic strain of the membrane. The strain is caused by the V-like shape of the dimers, unique among membrane protein complexes, which induces a strong deformation in the surrounding membrane. The process of row formation is therefore not a result of direct protein-protein interactions or a specific lipid composition of the membrane. We further hypothesize that, once assembled, the ATP synthase dimer rows prime the inner mitochondrial membrane to develop folds and invaginations by causing macroscopic membrane ridges that ultimately become the edges of cristae. In this way, mitochondrial ATP synthases would contribute to the generation of a morphology that maximizes the surface area of the inner membrane, and thus ATP production. Finally, we outline key experiments that would be required to verify or refute this hypothesis.


Age-Dependent Oxidative Stress Elevates Arginase 1 and Uncoupled Nitric Oxide Synthesis in Skeletal Muscle of Aged Mice.
Aging is associated with reduced muscle mass (sarcopenia) and poor bone quality (osteoporosis), which together increase the incidence of falls and bone fractures. It is widely appreciated that aging triggers systemic oxidative stress, which can impair myoblast cell survival and differentiation. We previously reported that arginase plays an important role in oxidative stress-dependent bone loss. We hypothesized that arginase activity is dysregulated with aging in muscles and may be involved in muscle pathophysiology. To investigate this, we analyzed arginase activity and its expression in skeletal muscles of young and aged mice. We found that arginase activity and arginase 1 expression were significantly elevated in aged muscles. We also demonstrated that SOD2, GPx1, and NOX2 increased with age in skeletal muscle. Most importantly, we also demonstrated elevated levels of peroxynitrite formation and uncoupling of eNOS in aged muscles. Our in vitro studies using C2C12 myoblasts showed that the oxidative stress treatment increased arginase activity, decreased cell survival, and increased apoptotic markers. These effects were reversed by treatment with an arginase inhibitor, 2(S)-amino-6-boronohexanoic acid (ABH). Our study provides strong evidence that L-arginine metabolism is altered in aged muscle and that arginase inhibition could be used as a novel therapeutic target for age-related muscle complications.


Convalescent plasma for COVID-19: male gender, older age and hospitalisation associated with high neutralising antibody levels, England, 22 April to 12 May 2020.
We analysed factors associated with neutralising antibody levels in 330 convalescent plasma donors. Women and younger donors were more likely not to have measurable neutralising antibodies, while higher antibody levels were observed in men, in older donors and in those who had been hospitalised. These data will be of value in the timely recruitment of convalescent plasma donors most likely to have high levels of neutralising antibodies for ongoing studies investigating its effectiveness.


Prioritization of high-cost new drugs for HCV: making sustainability ethical.
Hepatitis C virus (HCV) infection is a major health problem worldwide. Chronic HCV infection may in the long run cause cirrhosis, hepatic decompensation and hepatocellular carcinoma, with an ultimate disease burden of at least 350,000 deaths per year worldwide. The new generation of highly effective direct acting antivirals (DAA) to treat HCV infection brings major promises to infected patients in terms of exceedingly high rates of sustained virological response (SVR) but also of tolerability, allowing even the sickest patients to be treated. Even in the face of the excellent safety and efficacy and wide theoretical applicability of these regimens, their introduction is currently facing cost and access issues denying their use to many patients in need. Health systems in all countries are facing a huge problem of distributive justice, since while they should guarantee individual rights, among which the right to health in its broader sense, therefore not limited to healing, but extended to quality of life, they must also grant equal access to the healthcare resources and keep the distribution system sustainable. In the face of a disease with a relatively unpredictable course, where many but not of all chronically infected will eventually die of liver disease, selective allocation of this costly resource is debatable. In most countries the favorite solution has been a stratification of patients for prioritization of treatment, which means allowing Interferon-free DAA treatment only in patients with advanced fibrosis or cirrhosis, while keeping on hold persons with lesser stages of liver disease. In this report, we will perform an ethical assessment addressing the issues linked to access to new therapies, prioritization and eligibility criteria, analyzing the meaning of the term "distributive justice" and the different approaches that can guide us (individualistic libertarianism, social utilitarianism and egalitarianism) on this specific matter. Even if over time the price of new DAA will be reduced through competition and eventual patent expiration, the phenomenon of high drug costs will go on in the next decades and we need adequate tools to face the problems of distributive justice that come with it.


Immune Profiling and Precision Medicine in Systemic Lupus Erythematosus.
Systemic lupus erythematosus (SLE) is an autoimmune disorder with a wide range of clinical symptoms. Enormous progress has been made in the immunological and genetic understanding of SLE. However, the biology of disease heterogeneity in SLE has remained largely unexplored. Human immune profiling studies, helped by recent technological advances especially in single-cell and "omics" analyses, are now shedding light on the cellular and molecular basis of clinical symptoms and disease flares in individual patients. Peripheral blood immunophenotyping analysis with flow cytometry or mass cytometry are identifying responsible cell subsets and markers characteristic of disease heterogeneity. Transcriptome analysis is discovering molecular networks responsible for disease activity, disease subtype and future relapse. In this review, we summarize recent advances in the immune profiling analysis of SLE patients and discuss how they will be used for future precision medicine.


Short-term effects of heat and cold on respiratory drug use. A time-series epidemiological study in A Coruña, Spain.
The consumption of medication, especially over-the-counter drugs, can reflect environmental exposure with a lesser degree of severity in terms of morbidity. The non-linear effects of maximum and minimum apparent temperature on respiratory drug sales in A Coruña from 2006 to 2010 were examined using a distributed lag nonlinear model. In particular, low apparent temperatures proved to be associated with increased sales of respiratory drugs. The strongest consistent risk estimates were found for minimum apparent temperatures in respiratory drug sales with an increase of 33.4% (95% CI, 12.5%-58.0%) when the temperature changed from 2.8°C to -1.4 °C. These findings may serve to guide the planning of public health interventions to predict and manage the health effects of exposure to the thermal environment for lower degrees of morbidity. More precisely, significant increases in the use of measured over-the-counter medication could be used to identify and anticipate influenza outbreaks due to a more sensitive degree of the data source.


High-throughput SNP-genotyping analysis of the relationships among Ponto-Caspian sturgeon species.
Legally certified sturgeon fisheries require population protection and conservation methods, including DNA tests to identify the source of valuable sturgeon roe. However, the available genetic data are insufficient to distinguish between different sturgeon populations, and are even unable to distinguish between some species. We performed high-throughput single-nucleotide polymorphism (SNP)-genotyping analysis on different populations of Russian (Acipenser gueldenstaedtii), Persian (A. persicus), and Siberian (A. baerii) sturgeon species from the Caspian Sea region (Volga and Ural Rivers), the Azov Sea, and two Siberian rivers. We found that Russian sturgeons from the Volga and Ural Rivers were essentially indistinguishable, but they differed from Russian sturgeons in the Azov Sea, and from Persian and Siberian sturgeons. We identified eight SNPs that were sufficient to distinguish these sturgeon populations with 80% confidence, and allowed the development of markers to distinguish sturgeon species. Finally, on the basis of our SNP data, we propose that the A. baerii-like mitochondrial DNA found in some Russian sturgeons from the Caspian Sea arose via an introgression event during the Pleistocene glaciation. In the present study, the high-throughput genotyping analysis of several sturgeon populations was performed. SNP markers for species identification were defined. The possible explanation of the baerii-like mitotype presence in some Russian sturgeons in the Caspian Sea was suggested.


Business entity selection: why it matters to healthcare practitioners--part I--Conceptual framework, sole proprietorships, and partnerships.
The Bureau of Labor statistics indicates only a 50% four-year survivability rate among businesses classified as "education and health services." Gaining knowledge of IRS business entities can result in cost savings, operational efficiency, reduced liability, and enhanced sustainability. Each entity has unique disadvan- tages, depending on size, diversity of ownership, desire to expand, and profitability. Business structures should be compatible with the organizational mission or vision statement, services and products, and professional codes of ethics. Healthcare reform will require greater business acumen. We have an ethical duty to disseminate and acquire the knowledge to properly establish and manage healthcare practices to ensure sustainable services that protect and serve the community.


Accurate adsorption energies of small molecules on oxide surfaces: CO-MgO(001).
A hybrid MP2:DFT + D optimization method is applied using a 3 × 3 × 2 cluster model (Mg9O9) embedded in a 4 × 4 × 4 slab model. The calculated CO-Mg(2+) distance is 248 pm, and the calculated CO frequency (blue) shift is 20 cm(-1), 6 cm(-1) larger than the experimental value. For the structure obtained, MP2 calculations with basis set extrapolation on a series of cluster models of increasing size are performed. Taking into account the difference in the periodic limit at the DFT + D level, 20.9 ± 0.7 kJ mol(-1) is obtained as the estimate for the full periodic MP2 limit for the energy of CO desorption from the MgO(001) surface. CCSD(T) corrections are evaluated for the Mg9O9 cluster model using an augmented double-zeta basis set. Basis set extension effects are examined for smaller models. For a loading of Θ = 1/8, the estimated CCSD(T) value is 21.0 ± 1.0 kJ mol(-1), which is 0.4 ± 1.0 kJ mol(-1) larger than the (electronic) desorption energy derived in this study from TPD desorption barriers reported in the literature.


Gait speed and handgrip strength as predictors of all-cause mortality and cardiovascular events in hemodialysis patients.
Low physical performance in patients undergoing maintenance hemodialysis is associated with a high mortality rate. We investigated the clinical relevance of gait speed and handgrip strength, the two most commonly used methods of assessing physical performance. We obtained data regarding gait speed and handgrip strength from 277 hemodialysis patients and evaluated their relationships with baseline parameters, mental health, plasma inflammatory markers, and major adverse clinical outcomes. Low physical performance was defined by the recommendations suggested by the Asian Working Group on Sarcopenia. The prevalence of low gait speed and handgrip strength was 28.2 and 44.8%, respectively. Old age, low serum albumin levels, high comorbidity index score, and impaired cognitive functions were associated with low physical performance. Patients with isolated low gait speed exhibited a general trend for worse quality of life than those with isolated low handgrip strength. Gait speed and handgrip strength showed very weak correlations with different determining factors (older age, the presence of diabetes, and lower serum albumin level for low gait speed, and lower body mass index and the presence of previous cardiovascular events for low handgrip strength). Patients with low gait speed and handgrip strength had elevated levels of plasma endocan and matrix metalloproteinase-7 and the highest risks for all-cause mortality and cardiovascular events among the groups (adjusted hazard ratio of 2.72, p = 0.024). Elderly patients with low gait speed and handgrip strength were at the highest risk for poor clinical outcomes. Gait speed and handgrip strength reflected distinctive aspects of patient characteristics and the use of both factors improved the prediction of adverse clinical outcomes in hemodialysis patients. Gait speed seems to be a better indicator of poor patient outcomes than is handgrip strength.


Survival following allogeneic transplant in patients with myelofibrosis.
Allogeneic hematopoietic cell transplantation (HCT) is the only curative therapy for myelofibrosis (MF). In this large multicenter retrospective study, overall survival (OS) in MF patients treated with allogeneic HCT (551 patients) and without HCT (non-HCT) (1377 patients) was analyzed with Cox proportional hazards model. Survival analysis stratified by the Dynamic International Prognostic Scoring System (DIPSS) revealed that the first year of treatment arm assignment, due to upfront risk of transplant-related mortality (TRM), HCT was associated with inferior OS compared with non-HCT (non-HCT vs HCT: DIPSS intermediate 1 [Int-1]: hazard ratio [HR] = 0.26, P < .0001; DIPSS-Int-2 and higher: HR, 0.39, P < .0001). Similarly, in the DIPSS low-risk MF group, due to upfront TRM risk, OS was superior with non-HCT therapies compared with HCT in the first-year post treatment arm assignment (HR, 0.16, P = .006). However, after 1 year, OS was not significantly different (HR, 1.38, P = .451). Beyond 1 year of treatment arm assignment, an OS advantage with HCT therapy in Int-1 and higher DIPSS score patients was observed (non-HCT vs HCT: DIPSS-Int-1: HR, 2.64, P < .0001; DIPSS-Int-2 and higher: HR, 2.55, P < .0001). In conclusion, long-term OS advantage with HCT was observed for patients with Int-1 or higher risk MF, but at the cost of early TRM. The magnitude of OS benefit with HCT increased as DIPSS risk score increased and became apparent with longer follow-up.


Can we reduce health inequalities? An analysis of the English strategy (1997-2010).
England was the first European country to pursue a systematic policy to reduce socio-economic inequalities in health. This paper assesses whether this strategy has worked, and what lessons can be learnt. A review of documents was conducted, as well as an analysis of entry-points chosen, specific policies chosen, implementation of these policies, changes in intermediate outcomes, and changes in final health outcomes. Despite some partial successes, the strategy failed to reach its own targets, that is, a 10% reduction in inequalities in life expectancy and infant mortality. This is due to the fact that it did not address the most relevant entry-points, did not use effective policies and was not delivered at a large enough scale for achieving population-wide impacts. Health inequalities can only be reduced substantially if governments have a democratic mandate to make the necessary policy changes, if demonstrably effective policies can be developed, and if these policies are implemented on the scale needed to reach the overall targets.


Reptiles in Guadeloupe (French West Indies) are a reservoir of major human Salmonella enterica serovars.
The epidemiology of human Salmonella enterica infections in Guadeloupe (French West Indies) appears to be specific, with a higher prevalence of the subspecies enterica serovars Panama and Arechavaleta (Panama and Arechavaleta) than in other regions. A study was performed in Guadeloupe to identify the reservoir of Salmonella serovars by comparing their distribution in warm- and cold-blooded animals and in humans living in Guadeloupe and mainland France. Furthermore, a case-control study was conducted in 2012-2013 to identify the main epidemiologic risk factors for S. enterica infection among children under 15 years of age. Between June 2011 and December 2014, feces from 426 reptiles (322 anoles, 69 iguanas and 35 geckos) and 50 frogs distributed throughout Guadeloupe and nearby islands were investigated. The frequency of S. enterica carriage was 15.0% (n = 64) in reptiles but varied by species. The only significant risk factor for S. enterica infection was a more frequent presence of frogs in the houses of cases than in those of controls (P = 0.042); however, isolates were not collected. Panama and Arechavaleta were the two serovars most often recovered between 2005 and 2014 from humans living in Guadeloupe (24.5% (n = 174) and 11.5% (n = 82), respectively), which is in contrast to the low prevalence in mainland France (0.4%). Their presence at low frequencies in wild reptiles (4.6% (n = 3) and 3.1% (n = 2), respectively) and pigs (7.5% (n = 5) and 1.5% (n = 1), respectively) suggests a broad host range, and humans may be infected by indirect or direct contact with animals. These serovars are probably poorly adapted to humans and therefore cause more severe infections. The unusual subspecies houtenae serovar 43:z4,z32:- was a major subspecies in wild reptiles (24.6%, n = 16) and humans (9.4%, n = 67) but was not recovered from warm-blooded animals, suggesting that reptiles plays a key role in human infection.


An Automated Grading and Diagnosis System for Evaluation of Dry Eye Syndrome.
This article describes methods used to determine the severity of Dry Eye Syndrome (DES) based on Oxford Grading Schema (OGS) automatically by developing and applying a decider model. The number of dry punctate dots occurred on corneal surface after corneal fluorescein staining can be used as a diagnostic indicator of DES severity according to OGS; however, grading of DES severity exactly by carefully assessing these dots is a rather difficult task for humans. Taking into account that current methods are also subjectively dependent on the perception of the ophtalmologists coupled with the time and resource intensive requirements, enhanced diagnosis techniques would greatly contribute to clinical assessment of DES. Automated grading system proposed in this study utilizes image processing methods in order to provide more objective and reliable diagnostic results for DES. A total of 70 fluorescein-stained cornea images from 20 patients with mild, moderate, or severe DES (labeled by an ophthalmologist in the Keratoconus Center of Yildirim Beyazit University Ataturk Training and Research Hospital) used as the participants for the study. Correlations between the number of dry punctate dots and DES severity levels were determined. When automatically created scores and clinical scores were compared, the following measures were observed: Pearson's correlation value between the two was 0.981; Lin's Concordance Correlation Coefficients (CCC) was 0.980; and 95% confidence interval limites were 0.963 and 0.989. The automated DES grade was estimated from the regression fit and accordingly the unknown grade is calculated with the following formula: Gpred = 1.3244 log(Ndots) - 0.0612. The study has shown the viability and the utility of a highly successful automated DES diagnostic system based on OGS, which can be developed by working on the fluorescein-stained cornea images. Proper implemention of a computationally savvy and highly accurate classification system, can assist investigators to perform more objective and faster DES diagnoses in real-world scenerios.


Use of osteopathic manipulative treatment by Ohio osteopathic physicians in various specialties.
The authors mailed a survey designed to determine the use of osteopathic manipulative treatment (OMT) to the 2,318 active osteopathic physicians registered with the Ohio Osteopathic Association; 871 responses were received, for a response rate of 38%. Approximately 75% of the respondents had not or had rarely used OMT: 44% of these respondents did not use any OMT and 31% reported treating fewer than 10 patients with OMT during the week before the survey. Approximately 25% of the surveyed osteopathic physicians treated more than 10 patients with OMT, and about 6% of these treated more than 30 patients with OMT. Respondents represented 40 specialty disciplines. All of the osteopathic physicians in 17 specialties reported no OMT use, osteopathic physicians in 9 specialties reported using OMT for fewer than 10 patients during the previous week, and osteopathic physicians in 9 specialties reported using OMT for more than 10 patients during the previous week. Of the somatic dysfunctions listed in the survey, low back disorders were treated with OMT most often. Few osteopathic specialists used OMT for patients with asthma or chronic obstructive pulmonary disease. The data suggest that a great opportunity exists to increase the use of OMT by osteopathic physicians, especially those who are specialists.


Exploring the multi-sensory environment as a leisure resource for people with complex neurological disabilities.
The problems experienced by people with neurological disabilities create barriers to participation in leisure activities. Particular common features include cognitive deficits (attention, executive functioning, language and memory), functional difficulties (in activities of daily living), psychomotor impairment (dyspraxia and poor co-ordination), and behavioural difficulties (aggression, agitation, and wandering). The Multi-sensory environment (MSE) has the potential to accommodate some of these problems experienced by people with neurological disabilities and, as such, is a valuable leisure resource. Multi-sensory activity can address individual sensory needs, such as offering a stronger stimulus if initial attempts are unnoticed, and be offered alongside familiar activities and routines to enhance sensory awareness. If the complexity of the activity, individual needs, and MSE demands are matched, engagement in this activity may be achieved. The Pool Activity Level Occupational Profiling Tool can provide a protocol from which MSE activity may be facilitated. By using this tool the MSE fits well within the World Health Organisation International Classification of functioning, Disability and Health by facilitating participation and reducing environmental barriers.


Sensory analysis and consumer acceptance of 140 high-quality extra virgin olive oils.
Sensory analysis is a crucial tool for evaluating the quality of extra virgin olive oils. One aim of such an investigation is to verify if the sensory attributes themselves - which are strictly related to volatile and phenolic compounds - may permit the discrimination of high-quality products obtained by olives of different cultivars and/or grown in various regions. Moreover, a crucial topic is to investigate the interdependency between relevant parameters determining consumer acceptance and objective sensory characteristics evaluated by the panel test. By statistically analysing the sensory results, a grouping - but not discriminatory - effect was shown for some cultivars and some producing areas. The preference map shows that the most appreciated samples by consumers were situated in the direction of the 'ripe fruity' and 'sweet' axis and opposite to the 'bitter' and 'other attributes' (pungent, green fruity, freshly cut grass, green tomato, harmony, persistency) axis. Extra virgin olive oils produced from olives of the same cultivars and grown in the same areas shared similar sensorial attributes. Some differences in terms of expectation and interpretation of sensory characteristics of extra virgin olive oils might be present for consumers and panellists: most of the consumers appear unfamiliar with positive sensorial attributes, such as bitterness and pungency.


Genetic variability among elite popcorn lines based on molecular and morphoagronomic characteristics.
Knowledge of genetic diversity among genotypes and relationships among elite lines is of great importance for the development of breeding programs. Therefore, the objective of this study was to evaluate genetic variability based on the morphoagronomic and molecular characterization of 18 elite popcorn (Zea mays var. everta) lines to be used by Universidade Estadual de Maringá breeding programs. We used 31 microsatellite primers (widely distributed in the genome), and 16 morphological descriptors (including the resistance to maize white spot, common rust, polysora rust of maize, cercospora and leaf blights). The molecular data revealed variability among the lines, which were divided into four groups that were partially concordant with unweighted pair group method with arithmetic mean (UPMGA) and Bayesian clusters. The lines G3, G4, G11, and G13 exhibited favorable morphological characters and low disease incidence rates. The four groups were confirmed using the Gower distance in the UPGMA cluster; however, there was no association with the dissimilarity patterns obtained using the molecular data. The absence of a correlation suggests that both characterizations (morphoagronomic and molecular) are important for discriminating among elite popcorn lines.


Histone deacetylases 1 and 2 act in concert to promote the G1-to-S progression.
Histone deacetylases (HDACs) regulate gene expression by deacetylating histones and also modulate the acetylation of a number of nonhistone proteins, thus impinging on various cellular processes. Here, we analyzed the major class I enzymes HDAC1 and HDAC2 in primary mouse fibroblasts and in the B-cell lineage. Fibroblasts lacking both enzymes fail to proliferate in culture and exhibit a strong cell cycle block in the G1 phase that is associated with up-regulation of the CDK inhibitors p21(WAF1/CIP1) and p57(Kip2) and of the corresponding mRNAs. This regulation is direct, as in wild-type cells HDAC1 and HDAC2 are bound to the promoter regions of the p21 and p57 genes. Furthermore, analysis of the transcriptome and of histone modifications in mutant cells demonstrated that HDAC1 and HDAC2 have only partly overlapping roles. Next, we eliminated HDAC1 and HDAC2 in the B cells of conditionally targeted mice. We found that B-cell development strictly requires the presence of at least one of these enzymes: When both enzymes are ablated, B-cell development is blocked at an early stage, and the rare remaining pre-B cells show a block in G1 accompanied by the induction of apoptosis. In contrast, elimination of HDAC1 and HDAC2 in mature resting B cells has no negative impact, unless these cells are induced to proliferate. These results indicate that HDAC1 and HDAC2, by normally repressing the expression of p21 and p57, regulate the G1-to-S-phase transition of the cell cycle.


Inhibition of adenylyl cyclase by Gi alpha.
Evidence suggests that both alpha and beta gamma subunits of heterotrimeric guanine nucleotide-binding regulatory proteins (G proteins) inhibit adenylyl cyclase. Although type I adenylyl cyclase is inhibited directly by exogenous beta gamma, inhibition of adenylyl cyclase by Gi alpha has not been convincingly demonstrated in vitro. Concentration-dependent inhibition of adenylyl cyclases by purified Gi alpha subunits is described. Activated Gi alpha but not G(o) alpha was effective, and myristoylation of Gi alpha was required. The characteristics of the inhibitory effect were dependent on the type of adenylyl cyclase and the nature of the activator of the enzyme. The concentrations of Gi alpha required to inhibit adenylyl cyclase were substantially higher than those normally thought to be relevant physiologically. However, analysis indicates that these concentrations may be relevant and reasonable.


Changes in health-related quality of life (HRQoL) in a population-based sample of children and adolescents after 3 years of follow-up.
To assess changes in health-related quality of life (HRQoL) in children and adolescents over a 3-year period and to examine factors associated with change. A representative sample of Spanish children and adolescents aged 8-18 years and their parents completed the KIDSCREEN-52 questionnaire at baseline and again after 3 years. Data were also collected on gender, pubertal development (PD), and family socio-economic status (SES). Change in HRQOL over time was evaluated using effect sizes (ES). Generalized estimating equations (GEE) were used to analyze associations among changes in KIDSCREEN scores, socio-demographic factors, and pubertal development. Response rate at follow-up was 54% (n = 454). Overall, HRQoL worsened in eight out of the ten KIDSCREEN dimensions, with ES ranging from -0.10 (Moods and Emotions) to -0.34 (Psychological Well-being). The decrease was most marked in the intermediate age group (13-17 years of age at follow-up) and in girls. In the GEE models, pubertal development more strongly impacted changes in girls than in boys. In this representative, population-based sample of children and adolescents, moderate decrements in HRQoL were observed after 3 years. Changes were particularly important among girls and in relation to pubertal development. These results could provide useful reference data for other longitudinal studies in population sub-groups.


Effects of chronic sodium depletion on canine brain renin and cathepsin D activities.
The activities of brain renin and cathepsin D were measured in brain regions of 10 dogs on a normal sodium intake (65 mEq Na+/day) and 10 other dogs placed on a low sodium diet (less than 4 mEq Na+/day) for 21 days and given a diuretic. The purpose of this study was twofold: to assess the effect of sodium depletion on brain renin activity; and to assess in the same regions alterations in brain renin and cathepsin D activities. Sodium depletion caused a ninefold increase in plasma renin activity, hemoconcentration, and hyponatremia. In the presence of marked hyperreninemia, the average cerebral renin activity was reduced significantly; the most pronounced changes occurred in the upper and lower brain-stem regions. Cerebrospinal fluid renin was increased by 30%, but this change was not significant in sodium-depleted dogs. There were no significant alterations in cathepsin D activity whether assessed in total or regional brain areas. These observations support the view that there is an inverse relationship between plasma and brain renin activity in chronically sodium-depleted dogs. Additionally, evidence is provided that brain renin activity is modified independently from cathepsin D activity.


Characteristics and applications of small, portable gaseous air pollution monitors.
Traditional approaches for measuring air quality based on fixed measurements are inadequate for personal exposure monitoring. To combat this issue, the use of small, portable gas-sensing air pollution monitoring technologies is increasing, with researchers and individuals employing portable and mobile methods to obtain more spatially and temporally representative air pollution data. However, many commercially available options are built for various applications and based on different technologies, assumptions, and limitations. A review of the monitor characteristics of small, gaseous monitors is missing from current scientific literature. A state-of-the-art review of small, portable monitors that measure ambient gaseous outdoor pollutants was developed to address broad trends during the last 5-10 years, and to help future experimenters interested in studying gaseous air pollutants choose monitors appropriate for their application and sampling needs. Trends in small, portable gaseous air pollution monitor uses and technologies were first identified and discussed in a review of literature. Next, searches of online databases were performed for articles containing specific information related to performance, characteristics, and use of such monitors that measure one or more of three criteria gaseous air pollutants: ozone, nitrogen dioxide, and carbon monoxide. All data were summarized into reference tables for comparison between applications, physical features, sensing capabilities, and costs of the devices. Recent portable monitoring trends are strongly related to associated applications and audiences. Fundamental research requires monitors with the best individual performance, and thus the highest cost technology. Monitor networking favors real-time capabilities and moderate cost for greater reproduction. Citizen science and crowdsourcing applications allow for lower-cost components; however important strengths and limitations for each application must be addressed or acknowledged for the given use.


Nervonoylceramide (C24:1Cer), a lipid biomarker for ocular irritants released from the 3D reconstructed human cornea-like epithelium, MCTT HCE™.
Due to invasive and painful procedures during in vivo rabbit eye irritation test, in vitro alternative methods have been widely investigated. Recently, 3D reconstructed human cornea-like epitheliums (RhCEs) garner a huge attention. RhCEs employ the tissue viability as a primary endpoint to determine ocular irritancy but additional biomarkers may improve its predictive capacity. Here, we explored lipid biomarkers for ocular irritants in MCTT HCE™ RhCE model. Three irritants; sodium lauryl sulfate, benzalkonium chloride and triton X-100 were selected to represent anionic, cationic and non-ionic detergent respectively. After treating MCTT HCE™ with irritants, the alteration of lipids in the treated tissues was examined with Nile Red staining, which revealed the depletion of corneal lipids. We further quantitated the release of ceramides and free fatty acids, major lipid components of cornea, into the medium during the post-treatment incubation, employing a sensitive UPLC-MS/MS method. Among 44 lipid species, nervonoylceramide (C24:1Cer) was found to be released commonly by all three irritants in a concentration-dependent manner. Tests with 10 additional reference substances further supported that C24:1Cer release was significantly correlated with viability. Examination of the genes involved in the biosynthetic pathway for C24:1Cer revealed that stearoylCoA desaturase (SCD) and elongase1 (ELOVL1) were upregulated, suggesting that lipids and related genes may be employed as biomarkers for ocular irritants.


Interrupted Blood Feeding in Ticks: Causes and Consequences.
Ticks are obligate hematophagous arthropods and act as vectors for a great variety of pathogens, including viruses, bacteria, protozoa, and helminths. Some tick-borne viruses, such as Powassan virus and tick-borne encephalitis virus, are transmissible within 15-60 min after tick attachment. However, a minimum of 3-24 h of tick attachment is necessary to effectively transmit bacterial agents such as Ehrlichia spp., Anaplasma spp., and Rickettsia spp. to a new host. Longer transmission periods were reported for Borrelia spp. and protozoans such as Babesia spp., which require a minimum duration of 24-48 h of tick attachment for maturation and migration of the pathogen. Laboratory observations indicate that the probability of transmission of tick-borne pathogens increases with the duration an infected tick is allowed to remain attached to the host. However, the transmission time may be shortened when partially fed infected ticks detach from their initial host and reattach to a new host, on which they complete their engorgement. For example, early transmission of tick-borne pathogens (e.g., Rickettsia rickettsii, Borrelia burgdorferi, and Brucella canis) and a significantly shorter transmission time were demonstrated in laboratory experiments by interrupted blood feeding. The relevance of such situations under field conditions remains poorly documented. In this review, we explore parameters of, and causes leading to, spontaneous interrupted feeding in nature, as well as the effects of this behavior on the minimum time required for transmission of tick-borne pathogens.