Populations, pathogens, and epidemic phases: closing the gap between theory and practice in the prevention of sexually transmitted diseases.
The phase specific model for the prevention and control of sexually transmitted infections (STI) offers new insights into the strategic planning of programmes. The model illustrates the importance of modifying the focus of prevention and control activities to different subpopulations as the epidemic evolves over time. However, the practical application of phase specific approaches will depend on an understanding of the variability and determinants in the trajectory by which STI epidemics progress through epidemic phases. This paper draws on empirical observations from diverse populations to explore the influence of sexual behaviour patterns in populations, the biological characteristics of STI pathogens, and the population-pathogen interactions in relation to epidemic trajectories. In addition, various approaches to the determination of epidemic phase are presented.


Comparison of wound culture and bronchial lavage in the severely burned child: implications for antimicrobial therapy.
The relationship of the burn wound flora to microbial pathogens in the tracheobronchial tree has important implications for antimicrobial therapy in the severely burned patient. Management of septic complications is bolstered by surveillance quantitative wound cultures (QWC) and bronchial lavage fluid (BLF) cultures. To compare the organisms present in BLF with those found in QWC and to determine if QWC can predict BLF results. Results of BLF cultures from all patients who underwent bronchial lavage from January 1, 1996, to December 31, 1996, at our institution were compared with QWC data from the same date. Criteria for a positive match included an identical antibiotic susceptibility pattern and biotype. Match rates were calculated qualitatively and quantitatively. In 30 (48%) of the 62 BLF cultures, there was a match between the organism identified in the BLF and the QWC. When strict quantitative criteria were applied, the match rate was only 9 (14%) of 62. Burn size and inhalation injury had no significant effect on match rate. Whereas the microbial pathogens were similar in the QWC and BLF, linear regression showed no value of QWC in predicting BLF culture results. The difference between qualitative and quantitative match rates suggests cross-colonization between the burn wound and tracheobronchial tree, but little to no cross-infection. The QWC and BLF cultures must be performed independently in determining antimicrobial specificity in the burned patient.


A single amino acid in coat protein of Pepper mild mottle virus determines its subcellular localization and the chlorosis symptom on leaves of pepper.
Pepper mild mottle virus (PMMoV) causes serious economic losses in pepper production in China. In a survey for viral diseases on pepper, two PMMoV isolates (named PMMoV-ZJ1 and PMMoV-ZJ2) were identified with different symptoms in Zhejiang province. Sequence alignment analysis suggested there were only four amino acid differences between the isolates: Val262Gly, Ile629Met and Ala1164Thr in the replicase, and Asp20Asn in the coat protein. Infectious cDNA clones of both isolates were constructed and shown to cause distinctive symptoms. Chlorosis symptoms appeared only on PMMoV-ZJ2-infected plants and the Asp20Asn substitution in the CP was shown to be responsible. Confocal assays revealed that the subcellular localization pattern of the two CPs was different, CP20Asp was mainly located at the cell periphery, whereas most CP20Asn located in the chloroplast. Thus, a single amino acid in the CP determined the chlorosis symptom, accompanied by an altered subcellular localization.


The influence of dispersal and diet breadth on patterns of genetic isolation by distance in phytophagous insects.
To determine the effects of dispersal ability and diet breadth on population-genetic structure, we reviewed the allozyme literature and estimated genetic isolation by distance (IBD) for 43 species/host races of phytophagous insects. Subsequently, we tested two opposing hypotheses regarding the influence of dispersal ability on IBD: that IBD slopes do not vary with mobility, but that intercepts increase with mobility, and, alternatively, that IBD slopes vary with dispersal ability. We found that from tens of kilometers to more than 1,000 km, IBD is weak in sedentary and highly mobile species but pronounced in moderately mobile species. We attribute the weak IBD in strong dispersers to the homogenizing effects of gene flow, whereas in sedentary species, limited gene flow allows nearly all populations to diverge. In intermediate dispersers, genetic homogeneity is achieved at small spatial scales, but limited dispersal promotes genetic divergence over long distances. We also tested the hypothesis that IBD increases with decreasing diet breadth. We discovered no such pattern, casting doubt on the supposition that specialization promotes speciation by influencing population-genetic subdivision. Finally, we found that the number of populations is a more important consideration than the number of polymorphic loci in studies of IBD.


[Frequency of isolation of viable tuberculosis bacteria from patients with different forms of lung tuberculosis].
Tuberculosis mycobacteria isolated from patients with lung tuberculosis have been analyzed to define their viability. The character of bacterial excretions was studied in patients with different clinical forms and detection genesis of tuberculosis process. During 1998-2000 about 228 cases of growth absence after mycobacteria inoculation have been revealed under positive microscopy. When determining viability of tuberculosis mycobacteria, it was shown that in most cases patients with all the forms of the pathologic process, except for the fibrous-cavernysh form, excrete alive microorganisms. The considerably higher amount of alive tuberculosis mycobacteria was distinguished in people who had chronical or relapsed tuberculosis in comparison with newly diagnosed patients.


Robot-enhanced diabetes care for middle-aged and older adults living with diabetes in the community: A small sample size mixed-method evaluation.
This study assessed robot-enhanced healthcare in practical settings for the purpose of community diabetes care. A mixed method evaluation collected quantitative and qualitative data on diabetes patients over 45 (N = 30) and community pharmacists (N = 10). It took 15-20 min for the diabetes patients to interact with the robot. Before and after the interaction, questionnaires including a diabetes knowledge test, self-efficacy for diabetes, and feasibility of use of the robot was administered. In-depth interviews with both pharmacists and patients were also conducted. After interacting with the robot, a statistically significant improvement in diabetes knowledge (p < .001) and feasibility of the robot (p = .012) was found, but self-efficacy (p = .171) was not significantly improved. Five themes emerged from interviewing the diabetes patients: Theme 1: meets the needs of self-directed learning for the elderly; Theme 2: reduces alertness and creates comfortable interaction; Theme 3: vividness and richness enhance interaction opportunities; Theme 4: Robots are not without disadvantages, and Theme 5: Every person has unique tastes. Three themes emerged from interviewing pharmacists: Theme 1: Technology must meet the real needs of the patient; Theme 2: creates new services, and Theme 3: The use of robots must conform to real-life situations. Both the diabetes patients and the pharmacist reported more positive feedback on the robot-enhanced diabetes care than concerns. Self-directed learning, comfortable interaction, and vividness were the most focuses when using robot to enhance self-management for the patients. Pharmacists were most receptive to fit conforming with reality and creating new services.


Mutations in the rod outer segment membrane guanylate cyclase in a cone-rod dystrophy cause defects in calcium signaling.
Rod outer segment guanylate cyclase 1 (ROS-GC1) is a member of the subfamily of Ca(2+)-regulated membrane guanylate cyclases; and it is pivotal for vertebrate phototransduction. Two opposing regulatory modes control the activity of ROS-GC1. At nanomolar concentrations of Ca(2+), ROS-GC1 is activated by Ca(2+)-binding proteins named guanylate cyclase activating proteins (GCAPs). However, at micromolar concentrations of Ca(2+), ROS-GC1 is stimulated by S100beta [also named calcium-dependent (CD) GCAP]. This mode is not linked with phototransduction; instead, it is predicted to be involved in retinal synaptic activity. Two point mutations, E786D and R787C, in ROS-GC1 have been connected with cone-rod dystrophy (CORD6), with only one type of point mutation occurring in each family. The present study shows that the E786D mutation has no effect on the basal catalytic activity of ROS-GC1 and on its activation by GCAP1 and S100beta; however, the mutated cyclase becomes more activated by GCAP2. The R787C mutation has three consequences: (1) it causes major damage to the basal cyclase activity, (2) it makes the cyclase 5-fold more sensitive to activation by GCAP1; and 3) converts the cyclase into a form that is less sensitive to activation by GCAP2 and S100beta. Thus, the two CORD6-linked mutations in ROS-GC1, which occur at adjacent positions, result in vastly different biochemical phenotypes, and they are connected with very specific molecular defects in the Ca(2+) switching components of the cyclase. These defects, in turn, are proposed to have a profound effect on both the machinery of phototransduction and the retinal synapse. The study for the first time defines the biochemistry of CORD6 pathology in precise molecular terms.


Brominated flame retardants in the environment of Asia-Pacific: an overview of spatial and temporal trends.
In this paper, we summarize spatial and temporal trends of polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDs) in coastal and marine biota, and further assess human exposure to these brominated flame retardants (BFRs) in Asia-Pacific. The review is based mainly on the studies that were conducted in our laboratory and utilized samples archived in the environmental specimen bank (es-BANK) of Ehime University, Japan. The studies suggest that the target BFRs are ubiquitous in the environment of Asia-Pacific. Examination of spatial trends reveals that concentrations of these contaminants are relatively high in samples from Korea, South China and Japan. In general, the magnitude of environmental contamination by PBDEs in Asia-Pacific, as well as human exposure to these contaminants, seem to be comparable to or slightly higher than in Europe, but lower than in North America. Evaluation of temporal trends in concentrations of BFRs in marine mammals from the coastal waters of Japan and China showed drastic increase during the last 30 years. These changes in BFR levels in samples from Japan were in line with trends in production/use of the commercial formulations. Since the withdrawal of some PBDE products from the Japanese market in the 1990s, concentrations of HBCDs appear to exceed those of PBDEs, reflecting increasing usage of HBCDs over PBDEs. The increasing environmental contamination by BFRs in Chinese coastal waters indicates that contamination by BFRs has already become evident, even in developing countries. In view of the rising environmental levels and the high consumption volume of BFRs in Asia, further efforts should be made to monitor environmental contamination by these chemicals in order to identify sources and reduce emissions.


A low voltage-activated Ca2+ current mediates cytokine-induced pancreatic beta-cell death.
Insulin-dependent diabetes mellitus is characterized by the selective destruction of pancreatic beta-cells. Chronic treatment with cytokines induced a low voltage-activated (LVA) Ca2+ current in mouse beta-cells. The concomitant increase in the basal cytoplasmic free Ca2+ concentration ([Ca2+]i) was associated with DNA fragmentation and cell death. Antagonists of LVA Ca2+ channels prevented this elevation of basal [Ca2+]i and DNA fragmentation and reduced the percentage of cell death. Exposure to cytokines did not affect the profile of Ca2+ currents or basal [Ca2+]i in glucagon-secreting alpha-cells. An increased Ca2+ signal through LVA Ca2+ channels may thus be a key feature in cytokine-induced beta-cell destruction.


Modeling the Bistable Dynamics of the Innate Immune System.
The size of primary challenge with lipopolysaccharide induces changes in the innate immune cells phenotype between pro-inflammatory and pro-tolerant states when facing a secondary lipopolysaccharide challenge. To determine the molecular mechanisms governing this differential response, we propose a mathematical model for the interaction between three proteins involved in the immune cell decision making: IRAK-1, PI3K, and RelB. The mutual inhibition of IRAK-1 and PI3K in the model leads to bistable dynamics. By using the levels of RelB as indicative of strength of the immune responses, we connect the size of different primary lipopolysaccharide doses to the differential phenotypical outcomes following a secondary challenge. We further predict under what circumstances the primary LPS dose does not influence the response to a secondary challenge. Our results can be used to guide treatments for patients with either autoimmune disease or compromised immune system.


Prediction of irritancy in the human skin irritancy model and occupational setting.
It is of great importance to find ways to lower the incidence of chronic irritant contact dermatitis. In this process, it is crucial to have insight in the factors that can predict irritancy. This review offers a survey of recent findings in the field of skin irritancy testing, discussed in the context of renowned, older work. Extrinsic and intrinsic factors that may determine the outcome of irritancy testing in the human skin model are considered. In recent decades, there has been increasing interest in factors influencing the development of occupational dermatitis by means of prospective cohort studies. This promising new area of investigation is discussed separately.


Clinical science training at the University of Colorado: a measurement of trainee satisfaction.
Clinical science (CLSC) research education differs from basic science education in that many CLSC programs have an added goal of creating successful academicians. CLSC programs have expanded curricula that include teaching career development techniques, such as manuscript and grant writing, and helping young investigators establish successful mentor-mentee relationships. A group of K30 CLSC training program students coordinated a pilot survey to determine if the CLSC training programs at the University of Colorado were meeting the needs of the participants in both didactic courses and in other aspects of academic medicine, including research. The small group survey was conducted as part of a clinical outcomes assessment course. Opportunities for improvement in the CLSC training programs were explored based on the results. Of 117 CLSC training program participants surveyed, 56% responded. Overall, there was a positive improvement found for the didactic CLSC research constructs. Participants also reported success in manuscript publication and grant writing applications. The CLSC program, however, was not successful in coordinating faculty mentor support for student research projects for 78% of respondents. Once a mentoring relationship was established, students were satisfied with the mentoring they received. In general, CLSC trainees were satisfied that the K30 clinical research curriculum was meeting their needs. Many of the trainees were successful in developing academic skills during the program. Establishing a mentor relationship was the missing ingredient within the K30 CLSC training program. This may be an important component that should be considered when developing programs to create the next generation of clinician-scientists.


Immunotherapy of cancer with genetically modified tumor vaccines.
Distant metastasis is the major cause for therapeutic failures in Clinical oncology. Active immunotherapy in the adjuvant setting of patients with low tumor volume would contribute to further reduction of the remaining tumor and establish long-lasting immunity that could protect the patient from recurrence of disease. Studies employing rodent tumor models with little or no intrinsic immunogenicity have shown that genetically modified tumor cell preparations consisting of irradiated tumor cells transduced with and expressing cytokines, such as interleukin-2 (IL-2), IL-6, interferon-gamma (IFNgamma), or granulocyte-macrophage colony stimulating factor (GM-CSF), or co-stimulatory molecules, such as B7-I, were capable of inducing the regression of preexisting tumors and cure animals from their disease. Moreover, in some instances, the cured animals have retained immunological memory, as indicated by the fact that animals have resisted a second challenge with the parental tumor cells. Induction of potent immune responses in tumor-bearing animals against nonimmunogenic or weakly immunogenic tumors supports the view that active immunization of cancer patients deserves consideration despite lack of demonstrable immunogenicity in many human tumors.


A novel, fast-growing Borrelia sp. isolated from the hard tick Hyalomma aegyptium in Turkey.
A novel, fast-growing spirochaete was isolated from the hard tick Hyalomma aegyptium (family Ixodidae, subfamily Metastriata) using Barbour-Stoenner-Kelly (BSK) II medium. Tick samples were taken during the summer of 2000 from the Istanbul area in northwestern Turkey. Sixty-seven of 153 adults (44%) and 72 of 185 nymphs (39%) were infected with the novel spirochaete, whereas none of the 20 larvae examined were infected. The optimal growth temperature of the spirochaete in BSK II medium was 34-37 degrees C, and it could grow at 39 degrees C. Doubling times at 34 and 37 degrees C were 5.3 and 5.1 h, respectively. Six pure cultures of the spirochaete were obtained and characterized by microscopic observation, sequence analysis of the flagellin gene (flaB), SDS-PAGE and Western blotting. The spirochaete was morphologically similar to those of the genus Borrelia and contained a 41 kDa protein reactive with mAb H9724 specific to the flagellin of a Borrelia species. Polyclonal antibody raised to this spirochaete reacted with several antigen bands, whereas no bands were detected with Borrelia burgdorferi, Borrelia hermsii, Borrelia turicatae and Borrelia parkeri. The flaB sequences of the six isolates showed high similarity, with sequence similarity values ranging from 99.2 to 100%; however, the similarity of the isolates' flaB sequences to those of the Lyme-disease-related Borrelia and relapsing-fever-associated Borrelia species was less than 90%. These findings suggest that the unique spirochaete is a member of the genus Borrelia, and differs from previously described Borrelia species.


A Qualitative Investigation of Return to Sport After Arthroscopic Bankart Repair: Beyond Stability.
Arthroscopic shoulder stabilization is known to have excellent functional results, but many patients do not return to their preinjury level of sport, with return to play rates reported between 48% and 100% despite good outcome scores. To understand specific subjective psychosocial factors influencing a patient's decision to return to sport after arthroscopic shoulder stabilization. Case series; Level of evidence, 4. Semistructured qualitative interviews were conducted with patients aged 18 to 40 years who had undergone primary arthroscopic shoulder stabilization and had a minimum 2-year follow-up. All patients participated in sport before surgery without any further revision operations or shoulder injuries. Qualitative data analysis was performed in accordance with the Strauss and Corbin theory to derive codes, categories, and themes. Preinjury and current sport participation was defined by type, level of competition, and the Brophy/Marx shoulder activity score. Patient-reported pain and shoulder function were also obtained. A total of 25 patients were interviewed, revealing that fear of reinjury, shifts in priority, mood, social support, and self-motivation were found to greatly influence the decision to return to sport both in patients who had and had not returned to their preinjury level of play. Patients also described fear of sporting incompetence, self-awareness issues, recommendations from physical therapists, and degree of confidence as less common considerations affecting their return to sport. In spite of excellent functional outcomes, extrinsic and intrinsic factors such as competing interests, kinesiophobia, age, and internal stressors and motivators can have a major effect on a patient's decision to return to sport after arthroscopic shoulder stabilization. The qualitative methods used in this study provide a unique patient-derived perspective into postoperative recovery and highlight the necessity to recognize and address subjective and psychosocial factors rather than objective functional outcome scores alone as contributing to a patient's decision to return to play.


[Reorganization of the protein composition of the nuclear matrix of hepatoma cells after inhibition of DNA synthesis with novobiocin].
The nuclear matrix of Zajdela hepatoma cells, in which DNA synthesis was blocked by novobiocin, contained 2.5-3.0 times more DNA and protein not dissociating in 2 M NaCl than the nuclear matrix of control cells. Chromatography of nuclear matrix preparations on Sepharose 2B-CL resulted in isolation of tightly bound DNA-protein complexes which did not dissociate in 8 M urea or 0.1% SDS. Subsequent elution of DNA-protein complexes on a hydroxylapatite column with a buffer containing 4 M guanidine hydrochloride and 5 M urea caused partial dissociation of the complexes. Electrophoretic analysis revealed essential changes in the composition of proteins DNA-protein complexes of hepatoma cells nuclear matrix during inhibition of DNA synthesis.


The evolution of a G1/S transcriptional network in yeasts.
The G1-to-S cell cycle transition is promoted by the periodic expression of a large set of genes. In Saccharomyces cerevisiae G1/S gene expression is regulated by two transcription factor (TF) complexes, the MBF and SBF, which bind to specific DNA sequences, the MCB and SCB, respectively. Despite extensive research little is known regarding the evolution of the G1/S transcription regulation including the co-evolution of the DNA binding domains with their respective DNA binding sequences. We have recently examined the co-evolution of the G1/S TF specificity through the systematic generation and examination of chimeric Mbp1/Swi4 TFs containing different orthologue DNA binding domains in S. cerevisiae (Hendler et al. in PLoS Genet 13:e1006778. doi: 10.1371/journal.pgen.1006778 , 2017). Here, we review the co-evolution of G1/S transcriptional network and discuss the evolutionary dynamics and specificity of the MBF-MCB and SBF-SCB interactions in different fungal species.


Adult Attention Deficit Hyperactivity Disorder and comorbidity.
Attention Deficit Hyperactivity Disorder (ADHD) is a chronic and incapacitating mental disorder characterized by overactivity, inattention and impulsiveness. The syndrome is usually diagnosed in childhood and it continues into adulthood. However, because the symptoms of ADHD frequently overlap with other categories of psychopathology, in adults it is often underdiagnosed or neglected, and left untreated. This paper emphasizes the importance of recognizing this syndrome in adulthood. To this end, we provide a review of the literature and four illustrative cases of our own. Appropriate treatment is also discussed. (Int J Psych Clin Pract 2000; 4:35-39).


Critical evaluation of the existing guidelines on mild traumatic brain injury.
The purpose of guidelines is to reduce practice variability, but they need to be evidence-based. We examine current mild traumatic brain injury guidelines, critique their basis in evidence and examine their variability in recommendations. A systematic search of the literature found 38,806 abstracts, with 41 guidelines. There were 18 sports-related guidelines, 13 related to admission policies, 12 related to imaging and 5 related to neuropsychological assessment. Some guidelines addressed several areas. Only 5 guidelines reported a methodology for the assembly of evidence used to develop the guideline. After appraising the guidelines against a validated index, we found that 3 of the 41 guidelines could be categorized as evidence-based. Two of these focused on paediatric patients and 1 on adult patients. Limited methodological quality in the current guidelines results in conflicting recommendations amongst them.


[Prevalence of obesity and altered lipid profile in university students].
Obesity is a serious public health problem because its association with the risk to develop various chronic diseases. Atherogenic dyslipidemia that often accompany obesity is also associated to the metabolic syndrome and to cardiovascular diseases. The transition from adolescence to young adulthood appears to be a period where major changes occur in the lifestyle which contributes to the development of obesity, however, little attention has been given to this transition stage. The inclination to adopt unhealthy behaviors which occurs during early adulthood may be increased on university students because their lifestyle, which is characterized by lack of time to eat a healthy diet, which can make them susceptible to obesity. To determine the prevalence of obesity and lipid levels abnormalities and their relationship in a group of university students. Transversal study of university students aged between 18 and 24 years. Body mass index, waist circumference and blood lipid profile where evaluated. Of the 620 students surveyed about one-third have either overweight or obesity. 86% of students had at least one alteration in the evaluated parameters. Lipid profile results show a high prevalence of minor alterations in levels, particularly in cholesterol linked to low density lipoproteins levels. University young students have a high prevalence of overweight and plasma lipid levels above the norm, but most are in the low-risk categories. It is necessary to establish early preventive measures aimed at promoting in the university student good eating habits and increased physical activity.


Provision of respiratory support compared to no respiratory support before cord clamping for preterm infants.
Placental transfusion (by means of delayed cord clamping (DCC), cord milking, or cord stripping) confers benefits for preterm infants. It is not known if providing respiratory support to preterm infants before cord clamping improves outcomes. To assess the efficacy and safety of respiratory support provided during DCC compared with no respiratory support during placental transfusion (in the form of DCC, milking, or stripping) in preterm infants immediately after delivery. We used the standard search strategy of Cochrane Neonatal to search the Cochrane Central Register of Controlled Trials (CENTRAL, 2017, Issue 5), MEDLINE via PubMed (1966 to 19 June 2017), Embase (1980 to 19 June 2017), and CINAHL (1982 to 19 June 2017). We also searched clinical trials databases, conference proceedings, and the reference lists of retrieved articles for randomized controlled trials and quasi-randomized trials. Randomized, cluster randomized, or quasi-randomized controlled trials enrolling preterm infants undergoing DCC, where one of the groups received respiratory support before cord clamping and the control group received no respiratory support before cord clamping. All review authors assisted with data collection, assessment, and extraction. Two review authors assessed the quality of evidence using the GRADE approach. We contacted study authors to request missing information. One study fulfilled the review criteria. In this study, 150 preterm infants of less than 32 weeks' gestation undergoing 60 second DCC were randomized to a group who received respiratory support in the form of continuous positive airway pressure (CPAP) or positive pressure ventilation during DCC and a group that did not receive respiratory support during the procedure. Mortality during hospital admission was not significantly different between groups with wide confidence intervals (CI) for magnitude of effect (risk ratio (RR) 1.67, 95% CI 0.41 to 6.73). The study did not report neurodevelopmental disability and death or disability at two to three years of age. There were no significant differences between groups in condition at birth (Apgar scores or intubation in the delivery room), use of inotropic agents (RR 1.25, CI 0.63 to 2.49), and receipt of blood transfusion (RR 1.03, 95% CI 0.70 to 1.54). In addition, there were no significant differences in the incidences of any intraventricular haemorrhage (RR 1.50, 95% CI 0.65 to 3.46) and severe intraventricular haemorrhage (RR 1.33, 95% CI 0.31 to 5.75). Several continuous variables were reported in subgroups depending on method of delivery. Unpublished data for each group as a whole was made available and showed peak haematocrit in the first 24 hours and duration of phototherapy did not differ significantly. Overall, the quality of evidence for several key neonatal outcomes (e.g. mortality and intraventricular haemorrhage) was low because of lack of precision with wide CIs. The results from one study with wide CIs for magnitude of effect do not provide evidence either for or against the use of respiratory support before clamping the umbilical cord. A greater body of evidence is required as many of the outcomes of interest to the review occurred infrequently. Similarly, the one included study cannot answer the question of whether the intervention is or is not harmful.


Application of electrospray mass spectrometry to the structural determination of glycosylphosphatidylinositol membrane anchors.
The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the degree of structural information that can be obtained by applying electrospray mass spectrometry and tandem mass spectrometry to permethylated GPI glycans prepared from a well-characterized GPI-anchored glycoprotein, the variant surface glycoprotein from Trypanosoma brucei. All GPI glycans contain a non-N-acetylated glucosamine residue, and permethylation leads to the formation of a fixed positive charge on the glycans, in the form of a quaternary amine. The permethylated glycans were detected as [M +- Na](2+-) ions, and tandem mass spectrometry of these ions produced substantial, albeit incomplete, structural information on the branching patterns and linkage types for various GPI glycoforms of the variant surface glycoprotein.


Quantification of regional left ventricular wall motion in newborns by color kinesis.
Normal values for regional left ventricular wall motion, although documented in adults, have not been reported in healthy newborns. This study prospectively evaluated global and segmental systolic and diastolic cardiac function by color kinesis in clinically asymptomatic healthy newborns. Eighty-eight asymptomatic infants who were less than 48 hours old were studied. Systolic and diastolic parameters of global and regional left ventricular function are reported as means +/- SD, medians, 5th and 95th percentiles to establish the normative values for newborns. The reported fractional area changes during systole and diastole are similar to the reported normal values for older subjects. Higher body surface area significantly correlated with an increased peak velocity during systole, and fractional area changes during filling of the lateral wall. Our report of left ventricular regional wall-motion characteristics of healthy newborns, as evaluated by color kinesis, may help in the objective evaluation and management of newborns suspected to have global or segmental ventricular dysfunction.


Beech roots are simultaneously colonized by multiple genets of the ectomycorrhizal fungus Laccaria amethystina clustered in two genetic groups.
In this study, we characterize and compare the genetic structure of aboveground and belowground populations of the ectomycorrhizal fungus Laccaria amethystina in an unmanaged mixed beech forest. Fruiting bodies and mycorrhizas of L. amethystina were mapped and collected in four plots in the Świętokrzyskie Mountains (Poland). A total of 563 fruiting bodies and 394 mycorrhizas were successfully genotyped using the rDNA IGS1 (intergenic spacer) and seven simple sequence repeat markers. We identified two different genetic clusters of L. amethystina in all of the plots, suggesting that a process of sympatric isolation may be occurring at a local scale. The proportion of individuals belonging to each cluster was similar among plots aboveground while it significantly differed belowground. Predominance of a given cluster could be explained by distinct host preferences or by priority effects and competition among genets. Both aboveground and belowground populations consisted of many intermingling small genets. Consequently, host trees were simultaneously colonized by many L. amethystina genets that may show different ecophysiological abilities. Our data showed that several genets may last for at least 1 year belowground and sustain into the next season. Ectomycorrhizal species reproducing by means of spores can form highly diverse and persistent belowground genets that may provide the host tree with higher resilience in a changing environment and enhance ecosystem performance.


Charge transport in organic donor-acceptor mixed-stack crystals: the role of nonlocal electron-phonon couplings.
The charge-transport properties in C8BTBT-FnTCNQ and DMQtT-F4TCNQ mixed-stack crystals have been investigated by means of density functional theory, molecular dynamics and kinetic Monte Carlo simulations. The super-exchange nature of charge transport in these crystals is elucidated by the Larsson partition-based electronic coupling method that was developed recently by us. Compared with hole transport, in addition to the donor HOMO-acceptor LUMO interaction, the interaction between the donor HOMO-1 and the acceptor LUMO will also make an important contribution to electron transport. Moreover, this additional interaction plays an opposite role and results in electron-dominant and hole-dominant transport in the C8BTBT-FnTCNQ and DMQtT-F4TCNQ crystals, respectively. Most importantly, our calculations point out that the nonlocal electron-phonon couplings are very weak and much smaller than the electronic couplings in all the studied crystals. This implies that the nonlocal couplings have little influence on charge transport. In contrast to the experimental measurements, the external reorganization energies are thus expected to play an essential role in determining charge carrier mobilities. These findings pave the way for rational design of high performance organic donor-acceptor mixed-stack semiconductors.


Interaction of the signalling pathways of insulin-like growth factor-I and sex steroids in the neuroendocrine hypothalamus.
Among the numerous endocrine signals that affect the central nervous system, sex steroids play an important role. It has been recently postulated that part of the effects of these hormones on the brain may be mediated by trophic factors, such as insulin-like growth factor I (IGF-I). Both estradiol and IGF-I increase the survival and differentiation of developing fetal rat hypothalamic neurons in culture. The effect of estradiol is blocked by the pure estrogen receptor antagonist ICI 182,780, by an antisense oligonucleotide to the estrogen receptor, and by an antisense oligonucleotide to IGF-I. In turn, the effect of IGF-I is blocked by ICI 182,780 and by the antisense oligonucleotide to the estrogen receptor. These findings indicate that estrogen-induced activation of the estrogen receptor in developing hypothalamic neurons requires the presence of IGF-I and that both estradiol and IGF-I use the estrogen receptor to mediate their trophic effects on hypothalamic cells. In vivo, sex steroids affect IGF-I levels in the endocrine hypothalamus. IGF-I levels in tanycytes, a specific subtype of glial cells present in the arcuate nucleus and median eminence, are sexually dimorphic in the rat, increase with the onset of puberty, and are regulated by perinatal and adult levels of sex steroids. These changes may be due to hormonal modifications of IGF-I uptake by tanycytes from blood or cerebrospinal fluid. Therefore, this type of glial cell appears to play a central role in the interaction of sex steroids and IGF-I in the hypothalamus.


Knowledge, attitude, and practices related to COVID-19 pandemic among social media users in J&K, India.
A series of measures have been suggested to reduce Covid-19 infection, including knowledge training for prevention and control, isolation, disinfection, classified protections at different degrees in infection areas, and protection of confirmed cases. We conducted this study with an aim to assess the knowledge, attitude and practice among the general population regarding COVID-19. This was a cross-sectional study carried out by the Department of Community Medicine, Government Medical College, Srinagar in the month of April 2020. The questionnaire had four segments to collect data regarding social-demographic details, knowledge regarding Covid-19, attitude and practice based questions. The questionnaire was shared via social media applications like face book and Whatsapp to reach the target population. Continuous variables were summarized as frequency and percentage. All the analysis was done using Microsoft Excel 2016. Among participants who responded, 1252 (82%) were in the age group of 18-40 years and 912 (60%) from urban areas. A total of 934 (61%) respondents had heard details on COVID-19 from the social media, 1358 (89%) knew all ways of coronavirus transmission, 602 (40%) felt that COVID-19 is a serious disease, 1184 (78%) responded that they totally agree with the lockdown decision, and 1296 (85%) responded that lockdown is helping in reducing the number of cases. The majority, i.e. 1318 (87%), followed advisories and reported washing hands with soap and water regularly, 1108 (73%) reported regularly wearing masks, 1344 (89%) reported following lockdown guidelines, and 1306 (87%) reported maintaining social distancing. The respondents exhibited good knowledge, positive attitude, and sensible practices regarding COVID-19. Our study showed that the respondents have exhibited good knowledge, positive attitude and sensible practices regarding covid-19 during the pandemic.


Framework for continuous quality improvement in the provision of pharmaceutical care.
Implementation of the continuous quality improvement (CQI) process by health-care organizations is described. CQI involves a chain of events: Improvement in quality leads to decreased costs because of less rework, fewer mistakes, fewer delays, and better use of people and materials; productivity improves; better quality and lower price facilitate capturing the market, staying in business, and providing more jobs. The implementation of CQI within an organization involves the development, standardization, and deployment of standard methods and the use of daily management, cross-functional management, policy management, departmental management, and quality control circles to maintain, improve, or innovate standard methods. An organization's standard methods, key indicators (indicators for measuring performance of the organization's key functions), and standards emanate from its vision and mission statements and midterm and short-term strategic plans. In a quality control audit, an agreed-upon method, as monitored through a key indicator, is used to measure a result against an established standard; when a problem is detected, a quality control diagnosis is conducted and the method is changed if necessary. Use of this process requires the creation of standard methods for all important aspects of the organization's services, including operations, clinical services, and training of personnel. Health-care managers can use the principles of CQI in assessing the quality of care provided.


No Intern Left Behind: Using a Dedicated Transition to Discipline Block Improves Resident Outcomes.
This article was migrated. The article was marked as recommended. Introduction: A program was developed for incoming PGY-1 residents using the Accreditation Council for Graduate Medical Education milestones ratings. This program detects critical deficiencies and works to correct them as early as possible. Methods: A one month period was used for this transition to discipline block to identify at-risk learners. The block utilized cased-based discussions, interactive lectures, simulations, and clinical core rotations. All activities were tied to milestones measures to recognize deficiencies and provided a goal to correct the individual's progression. Results: Interns that completed the transition to discipline block were compared to the most recent previous class at the same institution. The same number of individuals with critical deficits were found in each class at first milestones rating (4 deficits per class, p value 1.0). The intervention classes had critical deficiencies recognized earlier and all identified deficiencies were extinguished earlier. Medical knowledge as compared by In-Training Examination percentile scores improved (Pre-Intervention Mean Percentile 28.9, Post-Intervention Mean Percentile 49.5, p value 0.005). Discussion: A milestones-based transition to residency block identified critical deficiencies earlier, which allowed for earlier intervention and improvement in resident performance. A similar process may benefit other residency programs.


Protein kinase Cα and P-type Ca channel CaV2.1 in red blood cell calcium signalling.
Protein kinase Cα (PKCα) is activated by an increase in cytosolic Ca(2+) in red blood cells (RBCs). Previous work has suggested that PKCα directly stimulates the CaV2.1 channel, whereas other studies revealed that CaV2.1 is insensitive to activation by PKC. The aim of this study was to resolve this discrepancy. We performed experiments based on a single cell read-out of the intracellular Ca(2+) concentration in terms of Fluo-4 fluorescence intensity and phosphatidylserine exposure to the external membrane leaflet. Measurement modalities included flow cytometry and live cell imaging. Treatment of RBCs with phorbol 12-myristate 13-acetate (PMA) led to two distinct populations of cells with an increase in intracellular Ca(2+): a weak-responding and a strong-responding population. The EC50 of PMA for the number of cells with Ca(2+) elevation was 2.7±1.2 µM; for phosphatidylserine exposure to the external membrane surface, it was 2.8±0.5 µM; and for RBC haemolysis, it was 2.9±0.5 µM. Using pharmacological manipulation with the CaV2.1 inhibitor ω-agatoxin TK and the broad protein kinase C inhibitor Gö6983, we are able to show that there are two independent PMA-activated Ca(2+) entry processes: the first is independent of CaV2.1 and directly PKCα-activated, while the second is associated with a likely indirect activation of CaV2.1. Further studies using lysophosphatidic acid (LPA) as a stimulation agent have provided additional evidence that PKCα and CaV2.1 are not directly interconnected in a signalling chain. Although we provide evidence for a lack of interaction between PKCα and CaV2.1 in RBCs, further studies are required to decipher the signalling relationship between LPA, PKCα and CaV2.1.


Interaction of formamidine pesticides with the presynaptic alpha(2)-adrenoceptor regulating.
The effects of the formamidine pesticides amitraz and chlordimeform on the alpha(2)-adrenergic receptor subtype that mediates the release of [(3)H]noradrenaline by synaptosomes from rat hypothalami were studied. We initially characterized the presynaptic autoreceptor on noradrenergic nerve endings using selective antagonists. Yohimbine (a nonselective alpha(2) antagonist) and BRL 44408 (selective for subtypes alpha(2A)/alpha(2D)) diminished the inhibitory effect of xylazine on K(+)-evoked release of [(3)H]noradrenaline; the K(B) values were 481 and 154 nM, respectively. In contrast, prazosin (a selective alpha(2B)/alpha(2C) antagonist) did not modify the inhibitory effect of xylazine. These results indicate that the release of noradrenaline by noradrenergic nerve endings in the rat hypothalamus is regulated by alpha(2D)-adrenoceptors, a species variation of the human alpha(2A) subtype. We then assessed the effects of the two pesticides on the K(+)-evoked release of [(3)H]noradrenaline. Amitraz reduced release in a dose-dependent manner; the effect observed at the maximal concentration tested (10 microM) was 13.0 +/- 2.0% and it was reversed by yohimbine. Amitraz also diminished the inhibitory effects of the alpha(2)-adrenergic agonists clonidine and xylazine. Chlordimeform displayed no effects, possibly because the true active compound of this insecticide is its demethylated metabolite. Based on these findings we conclude that the formamidine pesticides act as partial agonists of presynaptic alpha(2D)-adrenergic receptors in the rat hypothalamus. This interaction may be responsible for the in vivo alterations in catecholaminergic regulation of cyclic variations in gonadotropin-releasing hormone (GnRH) secretion, which can have grave functional repercussions on the reproductive system of mammals exposed to these xenobiotics.


NeuroEPO plus (NeuralCIM®) in mild-to-moderate Alzheimer's clinical syndrome: the ATHENEA randomized clinical trial.
NeuroEPO plus is a recombinant human erythropoietin without erythropoietic activity and shorter plasma half-life due to its low sialic acid content. NeuroEPO plus prevents oxidative damage, neuroinflammation, apoptosis and cognitive deficit in an Alzheimer's disease (AD) models. The aim of this study was to assess efficacy and safety of neuroEPO plus. This was a double-blind, randomized, placebo-controlled, phase 2-3 trial involving participants ≥ 50 years of age with mild-to-moderate AD clinical syndrome. Participants were randomized in a 1:1:1 ratio to receive 0.5 or 1.0 mg of neuroEPO plus or placebo intranasally 3 times/week for 48 weeks. The primary outcome was change in the 11-item cognitive subscale of the AD Assessment Scale (ADAS-Cog11) score from baseline to 48 weeks (range, 0 to 70; higher scores indicate greater impairment). Secondary outcomes included CIBIC+, GDS, MoCA, NPI, Activities of Daily Living Scales, cerebral perfusion, and hippocampal volume. A total of 174 participants were enrolled and 170 were treated (57 in neuroEPO plus 0.5 mg, 56 in neuroEPO plus 1.0 mg and 57 in placebo group). Mean age, 74.0 years; 121 (71.2%) women and 85% completed the trial. The median change in ADAS-Cog11 score at 48 weeks was -3.0 (95% CI, -4.3 to -1.7) in the 0.5 mg neuroEPO plus group, -4.0 (95% CI, -5.9 to -2.1) in the 1.0 mg neuroEPO plus group and 4.0 (95% CI, 1.9 to 6.1) in the placebo group. The difference of neuroEPO plus 0.5 mg vs. placebo was 7.0 points (95% CI, 4.5-9.5) P = 0.000 and between the neuroEPO plus 1.0 mg vs. placebo was 8.0 points (95% CI, 5.2-10.8) P = 0.000. NeuroEPO plus treatment induced a statistically significant improvement in some of clinical secondary outcomes vs. placebo including CIBIC+, GDS, MoCA, NPI, and the brain perfusion. Among participants with mild-moderate Alzheimer's disease clinical syndrome, neuroEPO plus improved the cognitive evaluation at 48 weeks, with a very good safety profile. Larger trials are warranted to determine the efficacy and safety of neuroEPO plus in Alzheimer's disease. https://rpcec.sld.cu Identifier: RPCEC00000232.


Atrial fibrillation and dementia in a population-based study. The Rotterdam Study.
Atrial fibrillation is a frequent disorder in the elderly and a known risk factor for cerebrovascular stroke. We investigated the association of atrial fibrillation with dementia and cognitive impairment in a large cross-sectional, population-based study in the elderly. Of the 6584 participants in the Rotterdam Study aged 55 to 106 years, detailed information on dementia status and ECG abnormalities was available. Dementia was diagnosed in three phases. First, participants were screened. Screen-positive subjects were tested further. Those with possible dementia underwent an extensive diagnostic workup. Dementia and dementia subtypes were diagnosed according to prevailing criteria. Cognitive impairment was defined as a Mini-Mental State Examination test score of < 26 points for a nondemented subject. Atrial fibrillation was diagnosed in 195, dementia in 276, and cognitive impairment in 635 subjects. We found significant positive associations of atrial fibrillation with both dementia and impaired cognitive function (age- and sex-adjusted odds ratios, 2.3 [95% confidence interval, 1.4 to 3.7] and 1.7 [95% confidence interval, 1.2 to 2.5]), respectively). The strongest association was found not for vascular dementia but rather for Alzheimer's disease with cerebrovascular disease. The associations were stronger in women, and the relation with dementia was more pronounced in the relatively younger elderly. A history of stroke in subjects with atrial fibrillation could not account for these associations. Dementia and subtypes Alzheimer's disease and vascular dementia may be related to atrial fibrillation even if no clinical stokes have occurred.


Experimental measurement and numerical analysis on resonant characteristics of cantilever plates for piezoceramic bimorphs.
Three experimental techniques are used in this study to access the resonant characteristics of piezoceramic bimorphs in parallel and series connections. These experimental methods, including the amplitude-fluctuation electronic speckle pattern interferometry (AF-ESPI), laser Doppler vibrometer-dynamic signal analyzer (LDV-DSA), and impedance analysis, are based on the measurement of full-field displacement, point-wise displacement, and electric impedance, respectively. Because the clear fringe patterns will be shown only at resonant frequencies, both the resonant frequencies and the corresponding vibration mode shapes are successfully obtained at the same time by the AF-ESPI method. LDV-DSA is used to determine the resonant frequencies of the vibration mode for out-of-plane motion. The impedance analysis is used to measure the resonant and antiresonant frequencies for in-plane motion. Although the out-of-plane mode is the dominant motion of piezoceramic bimorphs, it is found in this study that the amount of displacement for the in-plane motion in parallel connection is large enough to be measured by AF-ESPI and impedance. It is interesting to note that resonant frequencies of the specimen in parallel connection for the out-of-plane motion determined by LDV-DSA are the same as that for the in-plane motion obtained by impedance. Furthermore, both in-plane and out-of-plane mode shapes for the specimen in parallel connection are obtained in the same resonant frequency from the AF-ESPI method. It is concluded in this study that the particle motions of piezoceramic bimorphs for parallel connection in resonance are essentially three-dimensional. However, it is found that only out-of-plane vibration modes can be excited for the specimen in series connection. Numerical computations based on the finite-element method are presented, and the theoretical predicted results are compared with the experimental measurements. Good agreements between the experimental measured data and numerical calculated results are found for resonant frequencies and mode shapes of the piezoceramic bimorph.


Vanadium bioavailability in soils amended with blast furnace slag.
Blast furnace (BF) slags are commonly applied as soil amendments and in road fill material. In Sweden they are also naturally high in vanadium. The aim of this study was to assess the vanadium bioavailability in BF slags when applied to soil. Two soils were amended with up to 29% BF slag (containing 800 mg V kg(-1)) and equilibrated outdoors for 10 months before conducting a barley shoot growth assay. Additional soil samples were spiked with dissolved vanadate(V) for which assays were conducted two weeks (freshly spiked) and 10 months (aged) after spiking. The BF slag vanadium was dominated by vanadium(III) as shown by V K-edge XANES spectroscopy. In contrast, results obtained by HPLC-ICP-MS showed that vanadium(V), the most toxic vanadium species, was predominant in the soil solution. Barley shoot growth was not affected by the BF slag additions. This was likely due to limited dissolution of vanadium from the BF slag, preventing an increase of dissolved vanadium above toxic thresholds. The difference in vanadium bioavailability among treatments was explained by the vanadium concentration in the soil solution. It was concluded that the vanadium in BF slag is sparingly available. These findings should be of importance in environmental risk assessment.


Personalized dosimetry with intensification using 90Y-loaded glass microsphere radioembolization induces prolonged overall survival in hepatocellular carcinoma patients with portal vein thrombosis.
The objective of this study was to evaluate the response rate and survival of hepatocellular carcinoma portal vein thrombosis (PVT) patients treated with (90)Y-loaded glass microspheres using a personalized dosimetry and intensification concept. The microspheres were administered to 41 hepatocellular carcinoma PVT patients (main = 12; lobar/segmental = 29). (99m)Tc-macroaggregated albumin SPECT/CT quantitative analysis was used to calculate the tumor dose (TD), healthy injected liver dose (HILD), and injected liver dose (ILD). Response was evaluated at 3 mo using the criteria of the European Association for the Study of the Liver, with CT follow-up lasting until disease progression or death. Survival was assessed using the Kaplan-Meier method. The mean injected activity was 3.1 ± 1.5 GBq, and mean ILD was 143 ± 49 Gy. When a TD threshold of 205 Gy was applied, (99m)Tc-macroaggregated albumin SPECT/CT achieved a 100% sensitivity and 90% overall accuracy (0 false-negatives; 4 false-positives) in response prediction. On the basis of TD and HILD values, 37% of patients received an intensification of the treatment (increased injected activity with the aim of achieving a TD ≥ 205 Gy and HILD < 120 Gy, applying an ILD > 150 Gy). This intensification resulted in a high response rate (85%) without increased liver toxicity of grade 3 or higher (6% vs. 12% in the patients who did not receive treatment intensification; not statistically significant). For the total 41 patients, median overall survival (OS) was 18 mo (95% confidence interval, 11-25 mo). For patients with a TD of less than 205 Gy, median OS was 4.3 mo (3.7-5 mo), versus 18.2 mo (8.5-28.7 mo) for those with a TD of 205 Gy or more (P = 0.005). Median OS was 20.9 mo for patients with a TD of 205 Gy or more and good PVT targeting (n = 36). OS was 12 mo (3 mo to ∞) for patients with main PVT, versus 21.5 mo (12-28.7 mo) for those with segmental or lobar PVT (not statistically significant). For the 5 patients with complete portal vein revascularization who underwent lobar hepatectomy, median OS was not reached yet exceeded 24.5 mo and was significantly higher than that of other patients (P = 0.0493). Using a (99m)Tc-macroaggregated albumin SPECT/CT personalized dosimetry and intensification concept with (90)Y-loaded glass microspheres induced prolonged OS for PVT patients as compared with the standard of care (sorafenib), without increasing liver toxicity. Prospective randomized studies are therefore warranted.


Vascular risk factors and longitudinal changes on brain MRI: the ARIC study.
To evaluate associations between vascular risk factors and changes in burden of infarcts, ventricular size (VS), sulcal widening (SW), and white matter hyperintensities (WMH) in an initially middle-aged, biracial cohort from the Atherosclerosis Risk in Communities (ARIC) study. Initial brain magnetic resonance (MR) scans and evaluations for vascular risk factors were performed in 1,812 ARIC participants in 1994-1995. In 2004-2006, 1,130 ARIC participants underwent repeat MR scans. MR scans were rated using a validated 9-point scale for VS, SW, and WMH. Infarcts were recorded. Multiple logistic regression analysis was used to assess associations between vascular risk factors and change between MR scans of one or more grades in VS, SW, WMH, or appearance of new infarcts, controlling for age, sex, and race. At baseline, the 1,112 participants with usable scans (385 black women, 200 black men, 304 white women, 223 white men) had a mean age of 61.7 ± 4.3 years. In adjusted models, diabetes at baseline was associated with incident infarcts (odds ratio [OR] 1.95, 95% confidence interval [CI] 1.29-2.95) and worsening SW (OR 2.10, 95% CI 1.36-3.24). Hypertension at baseline was associated with incident infarcts (OR 1.73, 95% CI 1.23-2.42). In subjects with the highest tertile of fasting blood sugar and systolic blood pressure at baseline, the risk of incident infarcts was 3.68 times higher (95% CI 1.89-7.19) than those in the lowest tertile for both. Both atrophic and ischemic imaging changes were driven by altered glycemic and blood pressure control beginning in midlife.


Survival Rates and Bone and Soft Tissue Level Changes Around One-Piece Dental Implants Placed with a Flapless or Flap Protocol: 8.5-Year Results.
The purpose of the current study was to determine the survival rates and to measure marginal bone changes and peri-implant conditions 8.5 years after placement of one-piece implants with an anodically oxidized surface (AOS). A total of 52 subjects who received a one-piece implant with an AOS using a flapless or flap protocol and completed a previous randomized clinical trial were contacted for a recall visit 8.5 years after implant placement (T8.5). Implant success and survival rates, probing pocket depth (PPD), presence of bleeding on probing (BoP), papilla level, and incidence of complications and peri-implant disease were assessed by a single, blinded examiner. A second blinded examiner evaluated marginal bone level changes. Results for 8.5 years were compared to those at the time of implant placement, implant loading (0.5 year), and 1 and 1.5 years follow-up. The results based on 28 patients who attended the follow-up visit (half had flapless and half a flap protocol) showed a 100% implant survival rate and a 96.4% implant success rate 8.5 years after implant placement using one-piece implants, with no difference in survival and success rates between the flapless and the flap protocol. During the same follow-up period, a significant increase in crestal bone height from 1.5 to 8.5 years was observed. Analysis suggested decreasing mean levels of bone loss with time (P < .001). Moreover, there was 0.8 to 1.0 mm of bone loss through year 1.5, which decreased to 0.3 mm at 8.5 years (P < .05). There was no statistically significant difference in PPD or BoP over time. Similar mean levels of PPD were found in flap and flapless groups (mean [SD] = 2.4 [0.3] and 2.2 [0.4] mm, respectively [P = .18]), as well as similar rates of BoP (22.8% vs 17.9%, respectively). Papilla levels increased during the first year after implant loading. However, there was little additional change between 1.5 and 8.5 years. A total of eight fractured porcelain crowns and three crown loosenings were reported. One-piece implants with an AOS showed high survival rates and stable marginal bone and periimplant soft tissue levels regardless of whether a flapless or flap protocol was used.


Quantitative Proteomics of Maternal Blood Plasma in Isolated Intrauterine Growth Restriction.
Intrauterine growth restriction (IUGR) remains a significant concern in modern obstetrics, linked to high neonatal health problems and even death, as well as childhood disability, affecting adult quality of life. The role of maternal and fetus adaptation during adverse pregnancy is still not completely understood. This study aimed to investigate the disturbance in biological processes associated with isolated IUGR via blood plasma proteomics. The levels of 125 maternal plasma proteins were quantified by liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM MS) with corresponding stable isotope-labeled peptide standards (SIS). Thirteen potential markers of IUGR (Gelsolin, Alpha-2-macroglobulin, Apolipoprotein A-IV, Apolipoprotein B-100, Apolipoprotein(a), Adiponectin, Complement C5, Apolipoprotein D, Alpha-1B-glycoprotein, Serum albumin, Fibronectin, Glutathione peroxidase 3, Lipopolysaccharide-binding protein) were found to be inter-connected in a protein-protein network. These proteins are involved in plasma lipoprotein assembly, remodeling, and clearance; lipid metabolism, especially cholesterol and phospholipids; hemostasis, including platelet degranulation; and immune system regulation. Additionally, 18 proteins were specific to a particular type of IUGR (early or late). Distinct patterns in the coagulation and fibrinolysis systems were observed between isolated early- and late-onset IUGR. Our findings highlight the complex interplay of immune and coagulation factors in IUGR and the differences between early- and late-onset IUGR and other placenta-related conditions like PE. Understanding these mechanisms is crucial for developing targeted interventions and improving outcomes for pregnancies affected by IUGR.


The TRIB3 R84 variant is associated with increased carotid intima-media thickness in vivo and with enhanced MAPK signalling in human endothelial cells.
TRIB3, a mammalian tribbles homologue, affects insulin signalling and action by inhibiting Akt phosphorylation. A TRIB3 Q84R gain-of-function polymorphism has been associated with insulin resistance both in vitro and in vivo and with several atherosclerotic phenotypes, including increased carotid intima-media thickness (IMT). We wanted to replicate this latter association and, if so, to get deeper insights about the molecular mechanisms underlying the role of the TRIB3 Q84R polymorphism in atherosclerosis. in 430 Caucasians of European ancestry, carotid IMT was increased in QR (n = 116) and RR (n = 15) when compared with QQ (n = 299) subjects (P= 0.009), thus replicating similar data recently obtained among Asians. In human umbilical vein endothelial cells (HUVECs) naturally carrying the QQ genotype, 24 h insulin stimulation increased monocyte adhesion, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression, and mitogen-activated protein kinase (MAPK) kinase (MEK)-MAPK activation. Conversely, QR- and RR-HUVECs had increased unstimulated monocyte adhesion, VCAM-1 and ICAM-1 expression, and MEK-MAPK activation which did not increase further upon insulin stimulation. In addition, QQ-, QR-, and RR-HUVECs showed similar basal Akt phosphorylation and nitric oxide synthase activity which, however, were significantly increased by insulin only in QQ cells. the TRIB3 R4 variant is associated with increased carotid IMT also in Caucasians, thus replicating previous data obtained in Asians. In addition, in HUVECs, this variant is associated with unbalanced insulin signalling. This abnormality may favour vasoreactivity, intima-media thickening, and plaque formation and may, therefore, underlie the deleterious role exerted by the variant on the susceptibility to atherosclerosis.


Phospholipid morphologies on photochemically patterned silane monolayers.
We have studied the spreading of phospholipid vesicles on photochemically patterned n-octadecylsiloxane monolayers using epifluorescence and imaging ellipsometry measurements. Self-assembled monolayers of n-octadecylsiloxanes were patterned using short-wavelength ultraviolet radiation and a photomask to produce periodic arrays of patterned hydrophilic domains separated from hydrophobic surroundings. Exposing these patterned surfaces to a solution of small unilamellar vesicles of phospholipids and their mixtures resulted in a complex lipid layer morphology epitaxially reflecting the underlying pattern of hydrophilicity. The hydrophilic square regions of the photopatterned OTS monolayer reflected lipid bilayer formation, and the hydrophobic OTS residues supported lipid monolayers. We further observed the existence of a boundary region composed of a nonfluid lipid phase and a lipid-free moat at the interface between the lipid monolayer and bilayer morphologies spontaneously corralling the fluid bilayers. The outer-edge of the boundary region was found to be accessible for subsequent adsorption by proteins (e.g., streptavidin and BSA), but the inner-edge closer to the bilayer remained resistant to adsorption by protein or vesicles. Mechanistic implications of our results in terms of the effects of substrate topochemical character are discussed. Furthermore, our results provide a basis for the construction of complex biomembrane models, which exhibit fluidity barriers and differentiate membrane properties based on correspondence between lipid leaflets. We also envisage the use of this construct where two-dimensionally fluid, low-defect lipid layers serve as sacrificial resists for the deposition of protein and other material patterns.


Recall and recognition of verbal paired associates in early Alzheimer's disease.
The primary impairment in early Alzheimer's disease (AD) is encoding/consolidation, resulting from medial temporal lobe (MTL) pathology. AD patients perform poorly on cued-recall paired associate learning (PAL) tasks, which assess the ability of the MTLs to encode relational memory. Since encoding and retrieval processes are confounded within performance indexes on cued-recall PAL, its specificity for AD is limited. Recognition paradigms tend to show good specificity for AD, and are well tolerated, but are typically less sensitive than recall tasks. Associate-recognition is a novel PAL task requiring a combination of recall and recognition processes. We administered a verbal associate-recognition test and cued-recall analogue to 22 early AD patients and 55 elderly controls to compare their ability to discriminate these groups. Both paradigms used eight arbitrarily related word pairs (e.g., pool-teeth) with varying degrees of imageability. Associate-recognition was equally effective as the cued-recall analogue in discriminating the groups, and logistic regression demonstrated classification rates by both tasks were equivalent. These preliminary findings provide support for the clinical value of this recognition tool. Conceptually it has potential for greater specificity in informing neuropsychological diagnosis of AD in clinical samples but this requires further empirical support.


Hydration of ionic species studied by the reference interaction site model with a repulsive bridge correction.
We have tested the reference interaction site model (RISM) for the case of the hypernetted chain (HNC) and the partially linearized hypernetted chain (PLHNC) closures improved by a repulsive bridge correction (RBC) for ionic hydrated species. We have analyzed the efficiency of the RISM/HNC+RBC and RISM/PLHNC+RBC techniques for decomposition of the electrostatic and the nonpolar hydration energies on the energetic and the enthalpic parts for polyatomic ions when the repulsive bridge correction is treated as a thermodynamic perturbation, and investigate the repulsive bridge effect on the electrostatic potential induced by solvent on solute atoms. For a number of univalent and bivalent atomic ions, molecular cations, and anions, the method provides hydration energies deviating only by several percents from the experimental data. In most cases, the enthalpic contributions to the free energies are also close to the experimental results. The above models are able to satisfactory predict the hydration energies as well as the electrostatic potential around the ionic species. For univalent atomic ions, they also provide qualitative estimates of the Samoilov activation energies.


FDA-approved drugs and other compounds tested as inhibitors of human glutathione transferase P1-1.
Glutathione transferase P1-1 (GST P1-1) is often overexpressed in tumor cells and is regarded as a contributor to their drug resistance. Inhibitors of GST P1-1 are expected to counteract drug resistance and may therefore serve as adjuvants in the chemotherapy of cancer by increasing the efficacy of cytostatic drugs. Finding useful inhibitors among compounds used for other indications would be a shortcut to clinical applications and a search for GST P1-1 inhibitors among approved drugs and other compounds was therefore conducted. We tested 1040 FDA-approved compounds as inhibitors of the catalytic activity of purified human GST P1-1 in vitro. We identified chlorophyllide, merbromine, hexachlorophene, and ethacrynic acid as the most effective GST P1-1 inhibitors with IC50 values in the low micromolar range. For comparison, these compounds were even more potent in the inhibition of human GST A3-3, an enzyme implicated in steroid hormone biosynthesis. In distinction from the other inhibitors, which showed conventional inhibition patterns, the competitive inhibitor ethacrynic acid elicited strong kinetic cooperativity in the glutathione saturation of GST P1-1. Apparently, ethacrynic acid serves as an allosteric inhibitor of the enzyme. In their own right, the compounds investigated are less potent than desired for adjuvants in cancer chemotherapy, but the structures of the most potent inhibitors could serve as leads for the synthesis of more efficient adjuvants.


Ultrahigh-resolution optical coherence tomography in patients with decreased visual acuity after retinal detachment repair.
To assess microstructural changes in the retina that may explain incomplete visual recovery after anatomically successful repair of rhegmatogenous retinal detachments (RD) using ultrahigh-resolution optical coherence tomography (UHR OCT). Retrospective observational case series. Seventeen patients with decreased visual acuity after RD repair. Twelve patients had macula-involving and 5 had macula-sparing RDs. The UHR OCT prototype capable of approximately 3 mum axial resolution was developed for clinical use. The UHR OCT images through the center of the fovea in 17 patients with visual complaints after RD surgery were obtained. Patients were either postoperative patients from the New England Eye Center or tertiary referrals. Baseline visual acuity, preoperative lens status, location of retinal detachment, macular involvement, and postoperative visual acuity were recorded. The UHR OCT images after RD repair. The UHR OCT images were obtained 1 to 84 months (median, 5 months) postoperatively. The mean preoperative logarithm of the minimum angle of resolution (logMAR) visual acuity was 1.37 (Snellen equivalent, 20/390). The mean postoperative logMAR visual acuity was 0.48 (Snellen equivalent, 20/60). Anatomical abnormalities that were detected included distortion of the photoreceptor inner/outer segments (IS/OS) junction in 14 of 17 patients (82%), epiretinal membranes in 10 of 17 patients (59%), residual subretinal fluid in 3 of 17 patients (18%), and cystoid macular edema in 2 of 17 patients (12%). Of the 5 patients with preoperative macula-on detachments, 4 had distortion of the outer retina after RD repair. The higher resolution of UHR OCT facilitates imaging of the IS/OS junction. Therefore, UHR OCT is able to confirm prior histopathologic findings that damage to photoreceptor outer segments may occur as a consequence of retinal detachment. This may explain poor postoperative visual acuity in eyes with anatomically successful repair.


Utilization Rates of SGLT2 Inhibitors and GLP-1 Receptor Agonists and Their Facility-Level Variation Among Patients With Atherosclerotic Cardiovascular Disease and Type 2 Diabetes: Insights From the Department of Veterans Affairs.
There is mounting evidence regarding the cardiovascular benefits of sodium-glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide 1 receptor agonists (GLP-1 RA) among patients with atherosclerotic cardiovascular disease (ASCVD) and type 2 diabetes mellitus (T2DM). There is paucity of data assessing real-world practice patterns for these drug classes. We aimed to assess utilization rates of these drug classes and facility-level variation in their use. We used the nationwide Veterans Affairs (VA) health care system data set from 1 January 2020 to 31 December 2020 and included patients with established ASCVD and T2DM. Among these patients, we assessed the use of SGLT2i and GLP-1 RA and the facility-level variation in their use. Facility-level variation was computed using median rate ratios (MRR), a measure of likelihood that two random facilities differ in use of SGLT2i and GLP-1 RA in patients with ASCVD and T2DM. Among 537,980 patients with ASCVD and T2DM across 130 VA facilities, 11.2% of patients received an SGLT2i while 8.0% of patients received a GLP-1 RA. Patients receiving these cardioprotective glucose-lowering drug classes were on average younger and had a higher proportion of non-Hispanic Whites. Overall, median (10th-90th percentile) facility-level rates were 14.92% (9.31-22.50) for SGLT2i and 10.88% (4.44-17.07) for GLP-1 RA. There was significant facility-level variation among SGLT2i use-MRRunadjusted: 1.41 (95% CI 1.35-1.47) and MRRadjusted: 1.55 (95% CI 1.46 -1.63). Similar facility-level variation was observed for use of GLP-1 RA-MRRunadjusted: 1.34 (95% CI 1.29-1.38) and MRRadjusted: 1.78 (95% CI 1.65-1.90). Overall utilization rates of SGLT2i and GLP-1 RA among eligible patients are low, with significantly higher residual facility-level variation in the use of these drug classes. Our results suggest opportunities to optimize their use to prevent future adverse cardiovascular events among these patients.


In Vivo Selectivity and Localization of Reactive Oxygen Species (ROS) Induction by Osmium Anticancer Complexes That Circumvent Platinum Resistance.
Platinum drugs are widely used for cancer treatment. Other precious metals are promising, but their clinical progress depends on achieving different mechanisms of action to overcome Pt-resistance. Here, we evaluate 13 organo-Os complexes: 16-electron sulfonyl-diamine catalysts [(η6-arene)Os( N, N')], and 18-electron phenylazopyridine complexes [(η6-arene)Os( N, N')Cl/I]+ (arene = p-cymene, biphenyl, or terphenyl). Their antiproliferative activity does not depend on p21 or p53 status, unlike cisplatin, and their selective potency toward cancer cells involves the generation of reactive oxygen species. Evidence of such a mechanism of action has been found both in vitro and in vivo. This work appears to provide the first study of osmium complexes in the zebrafish model, which has been shown to closely model toxicity in humans. A fluorescent osmium complex, derived from a lead compound, was employed to confirm internalization of the complex, visualize in vivo distribution, and confirm colocalization with reactive oxygen species generated in zebrafish.


Development of Cardiometabolic Health indicators to advance the quality of spinal cord injury rehabilitation: SCI-High Project.
Context: Spinal cord injury or disease (SCI/D) leads to unchanged low-density lipoprotein and cholesterol, very low high-density lipoprotein a form of dyslipidemia and physical inactivity which combine to increase risk of morbidity and mortality from cardiometabolic disease. Herein, we describe the selection of structure, process and outcome indicators for adults in the first 18 months post-SCI/D rehabilitation admission. Methods: A Pan-Canadian Cardiometabolic Health Working Group was formed to develop a construct definition. Cardiometabolic risk factors were summarized in a Driver diagram. Release of the Paralyzed Veterans of America "Identification and Management of Cardiometabolic Risk after Spinal Cord Injury" and the International Scientific Exercise Guidelines: "Evidence-based scientific exercise guidelines for adults with spinal cord injury", informed the group's focus on prevention strategies to advance this Domain of rehabilitation admission. Results: The structure indicator identifies during rehabilitation the presence of appropriate time and resources for physical exercise prescription. Process indicators are lipid profile assessment at rehabilitation admission and documented exercise prescriptions prior to discharge. The outcome indicators track patient's knowledge retention regarding exercise prescription at discharge, current exercise adherence and lipid status 18 months after rehabilitation discharge. Conclusion: Routine national implementation of these indicators at the specified time points will enhance efforts to detect dyslipidemia and assure routine participation in endurance exercise. These indicators align with international initiatives to improve cardiometabolic health through interventions targeting modifiable risk factors specifically endurance exercising and optimal lipid profiles, crucial to augmenting cardiometabolic health after SCI/D.


A simple method for reducing cardiac output in the conscious lamb.
We have developed a method for reducing cardiac output in a controlled stepwise fashion using awake, intact, unsedated lambs. The method involves placing a balloon-tipped (Foley) catheter into the right atrium from a jugular vein isolated by a small neck incision. Systemic venous return and cardiac output are limited by balloon inflation. With each balloon inflation the animal reaches a new and stable cardiac output, which allows the measure of steady-state hemodynamic and metabolic variables. We have been able to decrease cardiac output to as low as 20% of the resting cardiac output and maintain a stable preparation. The reductions in cardiac output are quickly reversible by balloon deflation. Animal survival allows repeated study. We present data from five lambs studied between 26 and 36 days of age. Alterations in O2 consumption, O2 transport, O2 extraction, blood pressure, and arterial lactate concentration are examined in response to decrements in systemic blood flow and are consistent with changes seen in response to a reduction of cardiac output by other methods.


Brachybacterium endophyticum sp. nov., a novel endophytic actinobacterium isolated from bark of Scutellaria baicalensis Georgi.
A Gram-positive, aerobic, coccus-shaped, non-spore-forming actinobacterium, designated strain M1HQ-2T, was isolated from a surface-sterilized bark of Scutellaria baicalensis Georgi collected from Guizhou, China and tested using a polyphasic approach to determine its taxonomic position. Strain M1HQ-2T grew at 4-37 °C (optimum, 30 °C), pH 5.0-11.0 (pH 8.0) and in the presence of 0-15 % (w/v) NaCl (1-3 %). Substrate mycelia and aerial mycelia were not formed, and diffusible pigments were not observed on any media tested. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain M1HQ-2T belonged to the genus Brachybacterium and had the highest 16S rRNA gene sequence similarity of 97.6 % to Brachybacteriumsquillarum M-6-3T. Strain M1HQ-2T contained MK-7 as the dominant menaquinone. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The polar lipids profile of strain M1HQ-2T contained diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified lipid. The predominant fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content of strain M1HQ-2T was 71.0 mol%. The average nucleotide identity value between strain M1HQ-2T and type strain of Brachybacterium sacelli was 76.7 %. The estimated DNA-DNA hybridization value between strain M1HQ-2T and type strain of B. sacelli was 20.6 %. On the basis of phylogenetic analysis, chemotaxonomic characteristics and phenotypic data, strain M1HQ-2T represents a novel species of the genus Brachybacterium, for which the name Brachybacteriumendophyticum sp. nov. is proposed. The type strain is M1HQ-2T (=KCTC 49087T=CGMCC 1.16391T).


Characterization of the effects of n-butanol on the cell envelope of E. coli.
Biofuel alcohols have severe consequences on the microbial hosts used in their biosynthesis, which limits the productivity of the bioconversion. The cell envelope is one of the most strongly affected structures, in particular, as the external concentration of biofuels rises during biosynthesis. Damage to the cell envelope can have severe consequences, such as impairment of transport into and out of the cell; however, the nature of butanol-induced envelope damage has not been well characterized. In the present study, the effects of n-butanol on the cell envelope of Escherichia coli were investigated. Using enzyme and fluorescence-based assays, we observed that 1 % v/v n-butanol resulted in the release of lipopolysaccharides from the outer membrane of E. coli and caused 'leakiness' in both outer and inner membranes. Higher concentrations of n-butanol, within the range of 2-10 % (v/v), resulted in inner membrane protrusion through the peptidoglycan observed by characteristic blebs. The findings suggest that strategies for rational engineering of butanol-tolerant bacterial strains should take into account all components of the cell envelope.


Ocular torsional movements in humans with normal and abnormal ocular motility: Part II--Subjective observations.
A technique is described by which subjects, using a Maddox rod, can subjectively appreciate the dynamic torsional movements that occur at the end of forced head tilt. The subjective responses of a large series of subjects with normal ocular motility confirm my previous observations that at the end of ipsilateral head tilt the eye is undergoing an extorsional movement and at the end of contralateral head tilt, an intorsional movement. Observations made in patients with oblique dysfunction confirm that the extorsional movements are largely produced by the inferior oblique and the intorsional movements by the superior oblique. This is consistent with the classic observation that a partial compensatory intorsion occurs after ipsilateral tilt, and extorsion after contralateral tilt, however, it suggests that this compensatory torsion occurs by a different mechanism than is classically taught. Patients with dissociated vertical divergence appear to have an abnormal response of the oblique muscles to change in head tilt.


Quantitative structure-activity relationship study on affinity profile of a series of 1,8-naphthyridine antagonists toward bovine adenosine receptors.
The affinity profiles for the bovine adenosine receptors, A(1) and A(2A), of a series of 1,8-naphthyridine derivatives were quantitatively analyzed using physicochemical and structural parameters of the substituents, present at varying positions of the molecules. The derived significant correlation, for bovine A(1) receptor, suggested that a R(1) substituent having a higher van der Waals volume, a R(2) substituent being a hydrogen-bond donor and a R(3) substituent able to transmit a higher field effect are helpful in augmenting the pK(i) of a compound. Similarly the study, pertaining to bovine A(2A) receptor, revealed that a less bulky substituent at R(2) and a strong electron-withdrawing substituent at R(3) are desirable in improving the binding affinity of a compound while substituents at R(1) remain insignificant to any interaction.


Effect of ocular compression on intraocular pressure.
A prospective double-blind controlled study was undertaken to determine the effects of different compression times applied to soften the eye on normal subjects. Application of the Honan intraocular pressure reducer (HIPR) produced a significantly (p less than 0.02) greater decrease in intraocular pressure when applied for 40 minutes (14.3 mmHg SD = 1.7) when compared to a five-minute application (8.8 mmHg SD = 1.7). No untoward effects were noted in these patients. Physiologic effects of external ocular compression are discussed.


Exploring the impacts of safety culture on immigrants' vulnerability in non-motorized crashes: a cross-sectional study.
Pedestrians and cyclists are a vulnerable group of road users. Immigrants are disproportionally represented in pedestrian and cyclist crashes. We postulate that the mismatch in safety culture between countries of their origin and the U.S.A. contribute to their vulnerability in pedestrian and cyclist crashes. Over time, the differences may disappear and immigrants' traffic behavior gravitates toward those of native-borns. We describe this process as safety assimilation. Using the pedestrian and cyclist crash database in New York City between 2001 and 2003, we examined the effects of foreign-born population, their countries of origin, and time of entry into the USA on census tract-level pedestrian and cyclist crashes. We find that neighborhoods with a higher concentration of immigrants, especially those from Latin America, Eastern Europe, and Asia, have more crashes. Our results also exhibit a pattern of the hypothesized safety assimilation process. The study suggests a higher level of vulnerability of immigrants to pedestrian and cyclist crashes. We propose that targeted policies and programs need to be developed for immigrants of different countries of origin.


Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes.
Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D) cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-b2) and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-b2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro. These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.


Natural history of model organisms: The secret (group) life of Drosophila melanogaster larvae and why it matters to developmental ecology.
Model organisms such as Drosophila melanogaster have been key tools for advancing our fundamental and applied knowledge in biological and biomedical sciences. However, model organisms have become intertwined with the idea of controlled and stable laboratory environments, and their natural history has been overlooked.In holometabolous insects, lack of natural history information on larval ecology has precluded major advances in the field of developmental ecology, especially in terms of manipulations of population density early in life (i.e., larval density). This is because of relativistic and to some extent, arbitrary methodologies employed to manipulate larval densities in laboratory studies. As a result, these methodologies render comparisons between species impossible, precluding our understanding of macroevolutionary responses to population densities during development that can be derived from comparative studies.We recently proposed a new conceptual framework to address this issue, and here, we provide the first natural history investigation of Drosophila melanogaster larval density under such framework. First, we characterized the distribution of larval densities in a wild population of D. melanogaster using rotting apples as breeding substrate in a suburban area in Sweden.Next, we compiled the commonly used methodologies for manipulating larval densities in laboratory studies from the literature and found that the majority of laboratory studies identified did not manipulate larval densities below or above the densities observed in nature, suggesting that we have yet to study true life history and physiological responses to low and high population densities during D. melanogaster development.This is, to our knowledge, the first direct natural history account of larval density in nature for this model organism. Our study paves the way for a more integrated view of organismal biology which re-incorporates natural history of model organisms into hypothesis-driven research in developmental ecology.


Silica-induced pulmonary inflammation in rats: activation of NF-kappa B and its suppression by dexamethasone.
The goal of this study was to examine the relationship of the transcriptional regulatory factor nuclear factor-kappaB (NF-kappa B) to the early inflammatory events involved with silica exposure. Male F-344 rats received an intratracheal (i.t.) instillation of silica (100 mg/kg in a volume of 1 ml/kg) of saline. At 1, 3, 6, and 18 h postinstillation, and the rats were sacrificed and underwent bronchoalveolar lavage (BAL) for functional analysis of inflammation. Beginning at 1 h postinstillation, the silica-instilled (Si) rats displayed significant increases in neutrophils in BAL fluid compared to the saline controls. BAL cells from the Si group displayed a significant increase in luminol-dependent chemiluminescence (LDCL) compared to the controls. NF-kappa B activation was measurable at 3 h postinstillation, and this activation continued throughout the 18-h time course. Treatment with dexamethasone (5 mg/kg) at -3 h prior to silica instillation, at the time of instillation (0 h), and +1.5 h postinstillation resulted in both a reduction in NF-kappa B expression (by 70%) at 3 h postinstillation and corresponding reductions in LDCL, BAL cell count, and BAL neutrophils. These results show that activation of NF-kappa B is associated with silica-induced pulmonary inflammation, and the inhibition of its activation correlates temporally with suppression of inflammation.


The IMG/M data management and analysis system v.7: content updates and new features.
The Integrated Microbial Genomes & Microbiomes system (IMG/M: https://img.jgi.doe.gov/m/) at the Department of Energy (DOE) Joint Genome Institute (JGI) continues to provide support for users to perform comparative analysis of isolate and single cell genomes, metagenomes, and metatranscriptomes. In addition to datasets produced by the JGI, IMG v.7 also includes datasets imported from public sources such as NCBI Genbank, SRA, and the DOE National Microbiome Data Collaborative (NMDC), or submitted by external users. In the past couple years, we have continued our effort to help the user community by improving the annotation pipeline, upgrading the contents with new reference database versions, and adding new analysis functionalities such as advanced scaffold search, Average Nucleotide Identity (ANI) for high-quality metagenome bins, new cassette search, improved gene neighborhood display, and improvements to metatranscriptome data display and analysis. We also extended the collaboration and integration efforts with other DOE-funded projects such as NMDC and DOE Biology Knowledgebase (KBase).


Human brain single nucleus cell type enrichments in neurodegenerative diseases.
Single-cell RNA sequencing has opened a window into clarifying the complex underpinnings of disease, particularly in quantifying the relevance of tissue- and cell-type-specific gene expression. To identify the cell types and genes important to therapeutic target development across the neurodegenerative disease spectrum, we leveraged genome-wide association studies, recent single-cell sequencing data, and bulk expression studies in a diverse series of brain region tissues. We were able to identify significant immune-related cell types in the brain across three major neurodegenerative diseases: Alzheimer's disease, amyotrophic lateral sclerosis, and Parkinson's disease. Subsequently, putative roles of 30 fine-mapped loci implicating seven genes in multiple neurodegenerative diseases and their pathogenesis were identified. We have helped refine the genetic regions and cell types effected across multiple neurodegenerative diseases, helping focus future translational research efforts.


Tuning the Selectivity and Activity of Electrochemical Interfaces with Defective Graphene Oxide and Reduced Graphene Oxide.
Engineered solid-liquid interfaces will play an important role in the development of future energy storage and conversion (ESC) devices. In the present study, defective graphene oxide (GO) and reduced graphene oxide (rGO) structures were used as engineered interfaces to tune the selectivity and activity of Pt disk electrodes. GO was deposited on Pt electrodes via the Langmuir-Blodgett technique, which provided compact and uniform GO films, and these films were subsequently converted to rGO by thermal reduction. Electrochemical measurements revealed that both GO and rGO interfaces on Pt electrodes exhibit selectivity toward the oxygen reduction reaction (ORR), but they do not have an impact on the activity of the hydrogen oxidation reaction in acidic environments. Scanning transmission electron microscopy at atomic resolution, along with Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM), revealed possible diffusion sites for H2 and O2 gas molecules and functional groups relevant to the selectivity and activity of these surfaces. Based on these insights, rGO interfaces are further demonstrated to exhibit enhanced activity for the ORR in nonaqueous environments and demonstrate the power of our ex situ engineering approach for the development of next-generation ESC devices.


Diacylglycerol acyltransferase-2 (DGAT2) and monoacylglycerol acyltransferase-2 (MGAT2) interact to promote triacylglycerol synthesis.
Acyl CoA:1,2-diacylglycerol acyltransferase (DGAT)-2 is an integral membrane protein that catalyzes triacylglycerol (TG) synthesis using diacylglycerol and fatty acyl CoA as substrates. DGAT2 resides in the endoplasmic reticulum (ER), but when cells are incubated with fatty acids, DGAT2 interacts with lipid droplets presumably to catalyze localized TG synthesis for lipid droplet expansion. Previous studies have shown that DGAT2 interacts with proteins that synthesize its fatty acyl CoA substrates. In this study, we provide additional evidence that DGAT2 is present in a protein complex. Using a chemical cross-linker, disuccinimidyl suberate (DSS), we demonstrated that DGAT2 formed a dimer and was also part of a protein complex of ∼ 650 kDa, both in membranes and on lipid droplets. Using co-immunoprecipitation experiments and an in situ proximity ligation assay, we found that DGAT2 interacted with monoacylglycerol acyltransferase (MGAT)-2, an enzyme that catalyzes the synthesis of diacylglycerol. Deletion mutagenesis showed that the interaction with MGAT2 was dependent on the two transmembrane domains of DGAT2. No significant interaction of DGAT2 with lipin1, another enzyme that synthesizes diacylglycerol, could be detected. When co-expressed in cells, DGAT2 and MGAT2 co-localized in the ER and on lipid droplets. Co-expression also resulted in increased TG storage compared with expression of DGAT2 or MGAT2 alone. Incubating McArdle rat hepatoma RH7777 cells with 2-monoacylglycerol caused DGAT2 to translocate to lipid droplets. This also led to the formation of large cytosolic lipid droplets, characteristic of DGAT2, but not DGAT1, and indicated that DGAT2 can utilize monoacylglycerol-derived diacylglycerol. These findings suggest that the interaction of DGAT2 and MGAT2 serves to channel lipid substrates efficiently for TG biosynthesis.


[Comparison of clinical outcomes of blastocysts derived from non-top quality embryos and cleavage-stage high-quality embryos in frozen-thawed embryo transfer cycles].
To explore the developmental potential of embryos at different developmental days and provide evidence for blastocyst culture of non-top quality cleavage stage embryos in frozen-thawed embryo transfer (FET) cycles. The clinical data of 687 FET cycles were retrospectively analyzed. According to the embryo freezing time, the patients were divided into day 5 (D5) blastocyst group (n=87), day 6 (D6) blastocyst group (n=111) and day 3 cleavage-stage embryo (D3) group (n=489) with hormone replacement cycles or natural cycles for endometrial preparation. The clinical pregnancy rates, miscarriage rates, and implantation rates were compared between the 3 groups. The clinical pregnancy rate, miscarriage rate and implantation rate per transfer were 58.6%, 9.8%, and 42.9% in D5 group, 32.4%, 19.4%, and 23.3% in D6 group, and 44.9%, 16.4%, and 26.9% in D3 group, respectively. The clinical pregnancy rate and implantation rate were significantly higher in D5 group than in the other two groups (P<0.05). The D5 blastocysts derived from non-top quality D3 embryos after cryopreservation can have better clinical outcomes than those derived from D3 cleavage-stage embryos and D6 blastocysts, and are therefore a better option than D3 cleavage-stage embryos in FET cycles.