Characterization of a h-3 GnRH method for the measurement of GnRH binding-sites in human breast-cancer and breast-cancer cell-lines.
The peptide hormone, GnRH has been hypothesized to play a direct antitumoral role in certain kinds of breast cancer. This direct effect is considered to function via high affinity membrane receptors to GnRH that have been demonstrated in tumors. However, no routine assay for GnRH receptors exist limiting further research in elucidating the mechanisms of GnRH action during treatment and both the clinical usefulness of this measure and the development or refinement of GnRH treatments. Part of the problem has been that previously reported procedures require expensive, difficult (for routine clinical laboratories) iodination of specific GnRH analogs. The use of iodinated substrates present the additional problems of radioactive waste disposal and safety of laboratory personal. Further, the use of different analogs complicates the comparison of data between laboratories. We present an assay that utilizes tritiated natural GnRH in order to assay the native receptor binding and reduce both safety problems and cost of assay/waste disposal. The use of nitrocellulose filters treated overnight with 1% BSA resolved the problems of high blank values. Kinetic data demonstrate that this assay is sensitive, accurate and give results comparable with other methods. We also show that the membranes derived from the tumor and the cell cultures may be frozen at -80 degrees C for up to 90 days without affecting the results of the assay.


Synergistic effect of spexin and progesterone on pain sensitivity attenuation in ovariectomized rats.
Spexin is a central modulator of nociception. The aim of the present study was to investigate the effect of intra-hippocampal CA3 (IHCA3) injection of spexin and spexin-progesterone co-administration on pain sensitivity in ovariectomized rat. Thirty-five adult female rats were divided into five groups. Sham: the animals received injection of 0.5 μL ACSF by IHCA3. Experiments 1 and 2: the animals received injection of 0.5 μL of spexin bilaterally (10 and 30 nmol/rat respectively). Experiments 3 and 4: the animals received injection of 0.5 μL of spexin bilaterally (10 and 30 nmol/rat respectively) + subcutaneous (s.c.) injection of progesterone (5 mg/kg). Ovariectomy was performed in all groups to eliminate the effects of cyclic changes in the female rats. The formalin test (formalin 2.5%) was performed following the administration of spexin and progesterone. Results showed that bilateral injection of spexin in IHCA3 at both concentrations a significant (P < .05) decrease in the pain sensitivity in the two phases of formalin test. Similarly, the bilateral injection of spexin in IHCA3 at both concentrations following the s.c. injection of progesterone significantly (P < .05) decreases pain sensitivity in two phases of the formalin test. This pain attenuation due to the co-administration of spexin and progesterone was more potent than spexin-induced analgesia. According to the present results, spexin has a modulatory effect on pain sensitivity, which becomes more pronounced by progesterone administration.


Social-environmental phenotypes of rapid cystic fibrosis lung disease progression in adolescents and young adults living in the United States.
Cystic fibrosis (CF) is a genetic disease but is greatly impacted by non-genetic (social/environmental and stochastic) influences. Some people with CF experience rapid decline, a precipitous drop in lung function relative to patient- and/or center-level norms. Those who experience rapid decline in early adulthood, compared to adolescence, typically exhibit less severe clinical disease but greater loss of lung function. The extent to which timing and degree of rapid decline are informed by social and environmental determinants of health (geomarkers) is unknown. A longitudinal cohort study was performed (24,228 patients, aged 6-21 years) using the U.S. CF Foundation Patient Registry. Geomarkers at the ZIP Code Tabulation Area level measured air pollution/respiratory hazards, greenspace, crime, and socioeconomic deprivation. A composite score quantifying social-environmental adversity was created and used in covariate-adjusted functional principal component analysis, which was applied to cluster longitudinal lung function trajectories. Social-environmental phenotyping yielded three primary phenotypes that corresponded to early, middle, and late timing of peak decline in lung function over age. Geographic differences were related to distinct cultural and socioeconomic regions. Extent of peak decline, estimated as forced expiratory volume in 1 s of % predicted/year, ranged from 2.8 to 4.1 % predicted/year depending on social-environmental adversity. Middle decliners with increased social-environmental adversity experienced rapid decline 14.2 months earlier than their counterparts with lower social-environmental adversity, while timing was similar within other phenotypes. Early and middle decliners experienced mortality peaks during early adolescence and adulthood, respectively. While early decliners had the most severe CF lung disease, middle and late decliners lost more lung function. Higher social-environmental adversity associated with increased risk of rapid decline and mortality during young adulthood among middle decliners. This sub-phenotype may benefit from enhanced lung-function monitoring and personalized secondary environmental health interventions to mitigate chemical and non-chemical stressors.


Impact of extra-mitral valve cardiac involvement in patients with primary mitral regurgitation undergoing transcatheter edge-to-edge repair.
In the context of primary mitral regurgitation (PMR), the selection of patients for transcatheter edge-to-edge repair (TEER) does not include a systematic assessment of PMR-associated cardiac remodelling. We aimed to investigate the epidemiology and prognostic significance of different phenotypes of extra-mitral valve (MV) cardiac involvement in a large series of patients with PMR referred for TEER. The study included 654 patients from the multicentre Italian GIOTTO registry, stratified into groups according to extra-mitral valve (MV) cardiac involvement. The primary endpoint was all-cause death at 2-year follow-up. Patients with no cardiac involvement (NI; n=58), left heart involvement (LHI; n=343) and right heart involvement (RHI; n=253) were analysed. Acute technical success was achieved in 98% of patients. Kaplan-Meier curve analysis revealed significantly worse survival in patients with LHI and RHI (p=0.041). On multivariate Cox regression analysis, extra-MV cardiac involvement, haemoglobin level and technical success were independent predictors of the primary endpoint occurrence. Grading cardiac involvement may help refine risk stratification, since at least 1 group of extra-MV cardiac involvement represents in itself a negative predictor of midterm outcome.


Dysregulation of the Immune System in HIV/HCV-Coinfected Patients According to Liver Stiffness Status.
Advanced cirrhosis is related to alterations in immunity. We aimed to evaluate the levels of peripheral CD4⁺ T cells (Tregs) and plasma cytokine in patients coinfected with human immunodeficiency virus and hepatitis C virus (HIV/HCV) according to liver fibrosis stages [evaluated as liver stiffness measure (LSM)] and their linear relationship. We performed a cross-sectional study on 238 HIV/HCV-coinfected patients (119 had <12.5 kPa, 73 had 12.5⁻25 kPa, and 46 had >25 kPa). Peripheral T-cell subsets were phenotyped by flow cytometry, plasma biomarkers were assessed by multiplex immunoassays, and LSM was assessed by transient elastography. Results: We found HIV/HCV-coinfected patients had higher values of CD4⁺ Tregs (p < 0.001), memory Tregs (p ≤ 0.001), and plasma cytokine levels [IFN-γ (p ≤ 0.05) and IL-10 (p ≤ 0.01)] compared with healthy donors and HIV-monoinfected patients. In the multivariate analysis, higher LSM values were associated with reduced levels of IL-10 (adjusted arithmetic mean ratio (aAMR) = 0.83; p = 0.019), IL-2 (aAMR = 0.78; p = 0.017), TNF-α (aAMR = 0.67; p < 0.001), and IL-17A (aAMR = 0.75; p = 0.006). When we focus on HIV/HCV-coinfected patients analyzed by LSM strata, patients with ≥25 kPa had lower values of IL-2 (aAMR = 0.66; p = 0.021), TNF-α (aAMR = 0.565; p = 0.003), and IL-17A (aAMR = 0.58; p = 0.003) than patients with <12.5 kPa. HIV/HCV-coinfected patients showed an immunosuppressive profile compared to healthy controls and HIV-monoinfected patients. Additionally, HIV/HCV-coinfected patients with advanced cirrhosis (LSM ≥ 25 kPa) had the lowest plasma values of cytokines related to Th1 (IL-2 and TNF-α) and Th17 (IL-17A) response.


Development of surfactant coacervation in aqueous solution.
Coacervation is a phenomenon in which a colloidal dispersion separates into two immiscible liquid phases: a liquid rich in colloidal phase in equilibrium with another diluted liquid phase. Surfactant coacervation here refers to coacervation whose main components are surfactants with low molecular weights. Over the past two decades, surfactants have been greatly developed and studies on coacervation in systems of novel surfactants have been reported. This review summarizes the development of coacervation occurring in monomeric surfactants, one-head and two-tail surfactants, gemini surfactants and their mixtures. The effects of surfactant molecular structure and external conditions on critical conditions for coacervation, structures of precursors and coacervates, and their relationships are described. The effects of inorganic salts, alcohols and organic salts on surfactant coacervation are also reviewed.


Tobacco cessation education for dentists: an evaluation of the lecture format.
Dentists with tobacco cessation training perform more interventions, report increased self-efficacy, preparedness and fewer barriers than those without training. The aim of this study was to assess changes in knowledge, attitudes and behavior of dentists (CE group) at six months following presentation of a standardized tobacco cessation lecture module. These data were compared to those from age and gender-matched controls who did not receive training. The CE group was more likely to feel cessation was very important, score higher on knowledge questions, update tobacco use of continuing patients, ask former smokers about relapse and ask about daily consumption. The CE group was also more likely to discuss the personal relevance of quitting, roadblocks and setting quit dates, identify triggers, discuss pharmacotherapy and provide follow-up. These results suggest that group education appears to be a cost-efficient and effective method of teaching dentists about the latest methods of tobacco cessation.


Spontaneous activity of guinea pig ileum longitudinal muscle regulated by Ca(2+)-activated K+ channel.
Isolated guinea pig ileum displayed spontaneous contraction and fluctuation of membrane potential originating from the longitudinal muscle. Transmural stimulation of the enteric nerve plexus evoked contractions that were followed by lowered muscle tone and decreased spontaneous activity. The Na+ channel blocker tetrodotoxin, N-type Ca2+ channel blockers omega-conotoxins GVIA and MVIIA, the muscarinic receptor antagonist atropine, and inhibitory mediators alpha, beta-methylene-ATP and sodium nitroprusside all inhibited stimulation-evoked contraction while restoring spontaneous motility. L-type Ca2+ channel blockers nifedipine and calciseptine completely suppressed evoked and spontaneous contractions. Blockade of the large-conductance Ca(2+)-activated K+ (Kca) channel with charybdotoxin (but not of small-conductance Kca channel by apamin or of ATP-regulated K+ channel by glybenclamide) induced spikelike depolarization, inhibited nerve stimulation-evoked membrane hyperpolarization, and increased spontaneous activity. The results suggest that the large-conductance Kca channel is constitutively activated for modulations of spontaneous activity, and that muscle excitation, through elevation of Ca2+ levels, stimulates the large-conductance Kca channel to suppress spontaneous activity.


Establishing and evaluating a Balint group for fourth-year medical students at an Irish University.
The study was designed to establish and evaluate the impact of a 6-week Balint group on empathy and resilience in fourth-year medical students during their psychiatry rotation. This prospective study used the Jefferson Scale of Empathy - Student Version and the Brief Resilience Scale before and after 6-week Balint groups. Participating students also completed a qualitative assessment of their experience. Students who participated were enthusiastic regarding the value of Balint groups in promoting self-reflection and gaining insight into self- and patient-care dynamics. There was a significant difference in empathy scores pre- and post-Balint intervention. There was no significant difference in resilience scores. The establishment of a 6-week Balint group for fourth-year medical students was successful in increasing empathy. Students reported a positive view of Balint and its beneficial role in this study group.


Detection of Invasive Mosquito Vectors Using Environmental DNA (eDNA) from Water Samples.
Repeated introductions and spread of invasive mosquito species (IMS) have been recorded on a large scale these last decades worldwide. In this context, members of the mosquito genus Aedes can present serious risks to public health as they have or may develop vector competence for various viral diseases. While the Tiger mosquito (Aedes albopictus) is a well-known vector for e.g. dengue and chikungunya viruses, the Asian bush mosquito (Ae. j. japonicus) and Ae. koreicus have shown vector competence in the field and the laboratory for a number of viruses including dengue, West Nile fever and Japanese encephalitis. Early detection and identification is therefore crucial for successful eradication or control strategies. Traditional specific identification and monitoring of different and/or cryptic life stages of the invasive Aedes species based on morphological grounds may lead to misidentifications, and are problematic when extensive surveillance is needed. In this study, we developed, tested and applied an environmental DNA (eDNA) approach for the detection of three IMS, based on water samples collected in the field in several European countries. We compared real-time quantitative PCR (qPCR) assays specific for these three species and an eDNA metabarcoding approach with traditional sampling, and discussed the advantages and limitations of these methods. Detection probabilities for eDNA-based approaches were in most of the specific comparisons higher than for traditional survey and the results were congruent between both molecular methods, confirming the reliability and efficiency of alternative eDNA-based techniques for the early and unambiguous detection and surveillance of invasive mosquito vectors. The ease of water sampling procedures in the eDNA approach tested here allows the development of large-scale monitoring and surveillance programs of IMS, especially using citizen science projects.


Aminoglycoside redistribution phenomenon after hemodialysis: netilmicin and tobramycin.
The serum concentration time profile of netilmicin and tobramycin before, during, and after hemodialysis was assessed in 5 noninfected adult chronic hemodialysis patients. The pharmacokinetic profile was biexponential for both agents in the pre-hemodialysis period. The total body clearance of netilmicin was significantly greater than that of tobramycin (5.32 +/- 0.75 ml/min vs 3.66 +/- 1.00 ml/min; p less than 0.05). The hemodialysis clearances of netilmicin and tobramycin were similar (60.8 +/- 16.6 ml/min and 54.7 +/- 18.8 ml/min, respectively). Netilmicin and tobramycin serum concentrations increased significantly 10 minutes after cessation of hemodialysis and maximally rebounded to 38.3 +/- 16.2% and 18.3 +/- 3.0% at 1.7 +/- 0.3 hours and 1.9 +/- 0.7 hours, respectively. This phenomenon may be a primary contributor to the marked variability observed in the clinical pharmacokinetics of these agents in hemodialysis patients. These data suggest that clinical serum concentrations should not be drawn until two hours after hemodialysis.


Effects of a 1.5-Tesla static magnetic field on spermatogenesis and embryogenesis in mice.
There is a trend toward the use of higher magnetic field strengths in magnetic resonance imaging procedures. Considering this trend and the lack of consensus on the biologic effects of static magnetic fields, it is of considerable interest to examine the biologic effects of a 1.5-tesla (T) static magnetic field on spermatogenesis and embryogenesis in mice. Male and pregnant female Swiss Webster mice were exposed to a 1.5-T static magnetic field for 30 minutes. Effects on spermatogenesis in male mice were investigated by counting testicular spermheads and epididymal spermhead shape-abnormalities as a function of time after exposure. Pregnant female mice were exposed to the field at the two-cell embryo stage, sacrificed immediately, and the ability of these preimplantation embryos to mature into blastocysts was examined in vitro. Exposure to the static 1.5-T magnetic field caused a statistically significant reduction (15%) in testicular sperm on the 16th and 29th days after exposure. However, the increase in spermhead shape abnormalities above normal control values was minimal. A substantial effect was noted on the development of preimplantation embryos with a survival fraction of 0.56 compared with controls. A 30-minute exposure to a 1.5-T static magnetic field appears to cause some deleterious effects on spermatogenesis and embryogenesis in mice.


Microglia dynamic response and phenotype heterogeneity in neural regeneration following hypoxic-ischemic brain injury.
Hypoxic-ischemic brain injury poses a significant threat to the neural niche within the central nervous system. In response to this pathological process, microglia, as innate immune cells in the central nervous system, undergo rapid morphological, molecular and functional changes. Here, we comprehensively review these dynamic changes in microglial response to hypoxic-ischemic brain injury under pathological conditions, including stroke, chronic intermittent hypoxia and neonatal hypoxic-ischemic brain injury. We focus on the regulation of signaling pathways under hypoxic-ischemic brain injury and further describe the process of microenvironment remodeling and neural tissue regeneration mediated by microglia after hypoxic-ischemic injury.


Volumetric reduction in various cortical regions of elderly patients with early-onset and late-onset mania.
Few studies have examined alterations of the brain in elderly bipolar patients. As late-onset mania is associated with increased cerebrovascular morbidity and neurological damage compared with typical/early-onset mania, we investigated differences in the volume of various cortical regions between elderly patients with early-onset versus late-onset mania. We recruited 44 bipolar patients aged over 60 years, who underwent volumetric magnetic resonance imaging at 1.5 T. The analytic method is based on the hidden Markov random field model with an expectation-maximization algorithm. We determined the volume of each cortical region as a percentage of the total intracranial volume. The cutoff age for defining early versus late onset was 45 years. The study participants consisted of 25 patients with early-onset mania and 19 patients with late-onset mania; their mean ages were 65.7 years and 62.8 years, respectively. The demographic variables of the two groups were comparable. The volumes of the left caudate nucleus (p = 0.022) and left middle frontal gyrus (p = 0.013) were significantly greater and that of the right posterior cingulate gyrus (p = 0.019) was significantly smaller in the late-onset group. More patients with late-onset mania had comorbid cerebrovascular disease (p = 0.072). The right posterior cingulate gyrus is smaller and the left caudate nucleus and left middle frontal gyrus are larger in patients with late-onset mania compared with those with early-onset mania. Volumetric change in brain regions may vary in elderly bipolar patients with early and late-onset mania.


Nurse managers' experience with ethical issues in six government hospitals in Malaysia: a cross-sectional study.
Nurse managers have the burden of experiencing frequent ethical issues related to both their managerial and nursing care duties, according to previous international studies. However, no such study was published in Malaysia. The purpose of this study was to explore nurse managers' experience with ethical issues in six government hospitals in Malaysia including learning about the way they dealt with the issues. A cross-sectional study was conducted in August-September, 2010 involving 417 (69.2%) of total 603 nurse managers in the six Malaysian government hospitals. Data were collected using three-part self-administered questionnaire. Part I was regarding participants' demographics. Part II was about the frequency and areas of management where ethical issues were experienced, and scoring of the importance of 11 pre-identified ethical issues. Part III asked how they dealt with ethical issues in general; ways to deal with the 11 pre-identified ethical issues, and perceived stress level. Data were analyzed using descriptive statistics, cross-tabulations and Pearson's Chi-square. A total of 397 (95.2%) participants experienced ethical issues and 47.2% experienced them on weekly to daily basis. Experiencing ethical issues were not associated with areas of practice. Top area of management where ethical issues were encountered was "staff management", but "patient care" related ethical issues were rated as most important. Majority would "discuss with other nurses" in dealing generally with the issues. For pre-identified ethical issues regarding "patient care", "discuss with doctors" was preferred. Only 18.1% referred issues to "ethics committees" and 53.0% to the code of ethics. Nurse managers, regardless of their areas of practice, frequently experienced ethical issues. For dealing with these, team-approach needs to be emphasized. Proper understanding of the code of ethics is needed to provide basis for reasoning.


Topoisomerase II beta interacts with cohesin and CTCF at topological domain borders.
Type II DNA topoisomerases (TOP2) regulate DNA topology by generating transient double stranded breaks during replication and transcription. Topoisomerase II beta (TOP2B) facilitates rapid gene expression and functions at the later stages of development and differentiation. To gain new insight into the genome biology of TOP2B, we used proteomics (BioID), chromatin immunoprecipitation, and high-throughput chromosome conformation capture (Hi-C) to identify novel proximal TOP2B protein interactions and characterize the genomic landscape of TOP2B binding at base pair resolution. Our human TOP2B proximal protein interaction network included members of the cohesin complex and nucleolar proteins associated with rDNA biology. TOP2B associates with DNase I hypersensitivity sites, allele-specific transcription factor (TF) binding, and evolutionarily conserved TF binding sites on the mouse genome. Approximately half of all CTCF/cohesion-bound regions coincided with TOP2B binding. Base pair resolution ChIP-exo mapping of TOP2B, CTCF, and cohesin sites revealed a striking structural ordering of these proteins along the genome relative to the CTCF motif. These ordered TOP2B-CTCF-cohesin sites flank the boundaries of topologically associating domains (TADs) with TOP2B positioned externally and cohesin internally to the domain loop. TOP2B is positioned to solve topological problems at diverse cis-regulatory elements and its occupancy is a highly ordered and prevalent feature of CTCF/cohesin binding sites that flank TADs.


[Amphibians and reptiles in the swamps dominated by the palm Raphia taedigera (Arecaceae) in northeastern Costa Rica].
The herpetofauna that inhabits Caribbean Costa Rica has received considerable attention in the last two decades. This assemblage includes a total of 141 species of reptiles and 95 amphibians mostly distributed in tropical wet and moist lowland forests. While most information available came from primary and secondary forest sites, little is known about the amphibians and reptiles that inhabit more open habitats, such as wetlands and swamps. For instances, swaps dominated by the yolillo palm Raphia taedigera extend through much of the northeastern Caribbean coast of Costa Rica and eastern Nicaragua, but information about the herpetological community that uses such environments remains practically unknown. This situation reflects the little research conducted in such inhospitable environments. Here, we report the results of an intensive survey conducted to assess the herpetological community that inhabit R. taedigera palm-swamps. A total of 14 species of amphibians and 17 of reptiles have been recorded from these swamps. Amphibians and reptiles that inhabit yolillo swamps have wide distributions along much of Middle America and are considered common species throughout their range. In general, yolillo swamps are poor environments for herpetofauna: richness of reptiles and amphibians is almost two times higher in the adjacent forest than in the palm dominated swamps. Furthermore, most species observed in this swamps can be considered habitat generalists that are well adapted to the extreme conditions imposed by the changes in hydroperiods, reduce understory cover, low tree diversity and simple forest architecture of these environments. Despite similarities in the herpetofauna, it is clear that not all forest species use yolillo habitat, a characteristic that is discussed in terms of physical stress driven by the prolonged hydroperiod and reduced leaflitter in the ground, as these features drive habitat structure and herpetofaunal complexity. Our list of species using yolillo swamps is not definitive, and is likely to grow as more studies are conducted in this unexplored environment.


Metabolomic and biochemical analysis of mesocarp tissues from table grape berries with contrasting firmness reveals cell wall modifications associated to harvest and cold storage.
Tissue texture influences the grape berry consumers acceptance. We studied the biological differences between the inner and outer mesocarp tissues in hard and soft berries of table grapes cv NN107. Texture analysis revealed lower levels of firmness in the inner mesocarp as compared with the outer tissue. HPAEC-PAD analysis showed an increased abundance of cell wall monosaccharides in the inner mesocarp of harder berries at harvest. Immunohistochemical analysis displayed differences in homogalacturonan methylesterification and cell wall calcium between soft and hard berries. This last finding correlated with a differential abundance of calcium measured in the alcohol-insoluble residues (AIR) of the inner tissue of the hard berries. Analysis of abundance of polar metabolites suggested changes in cell wall carbon supply precursors, providing new clues in the identification of the biochemical factors that define the texture of the mesocarp of grape berries.


Direct integration of cell-free-synthesized connexin-43 into liposomes and hemichannel formation.
Proteoliposomes were directly prepared by synthesizing membrane proteins with the use of minimal protein synthesis factors isolated from Escherichia coli (the PURE system) in the presence of liposomes. Connexin-43 (Cx43), which is a water-insoluble integral membrane protein that forms a hexameric complex in membranes, was cotranslationally integrated with an essentially uniform orientation in liposomes. The addition of liposomes following protein expression (post-translational presence of liposomes) did not lead to the integration of Cx43 into the liposome membranes. The amount of integrated Cx43 increased as the liposome concentration increased. The presence of liposomes did not influence the total amount of synthesized Cx43. The Cx43 integrated into the liposome membranes formed open membrane pores. These results indicate that the liposomes act in a chaperone-like manner by preventing Cx43 from aggregating in solution, because of integration into the bilayer, and also by functionalization of the integrated Cx43 in the membrane. This is the first report that cell-free-synthesized water-insoluble membrane protein is directly integrated with a uniform orientation as a functional oligomer into liposome membranes. This simple proteoliposome preparation procedure should be a valuable approach for structural and functional studies of membrane proteins.


Spontaneous and aging-dependent development of arthritis in NADPH oxidase 2 deficiency through altered differentiation of CD11b+ and Th/Treg cells.
Emerging evidence indicates that NADPH oxidase (NOX) and its reactive oxygen species (ROS) products modulate a variety of cellular events, including proliferation, differentiation, and apoptosis. In this study, we investigated the functions of NOX2 and ROS in immune modulation using NOX2 knockout (KO) mice. Interestingly, NOX2 KO mice spontaneously developed arthritis with onset at 6-7 wk of age and high incidence (60%) at 15-18 wk of age. Arthritis severity in NOX2 KO mice was proportionally increased with age and higher in females than in males. Bone destruction was confirmed by microcomputed tomography scanning and histological analyses of joints. Inflammatory factors, including TNF-α, IL-1β, and RANKL, and serum level of anti-type II collagen IgG were significantly increased in NOX2 KO mice. In addition, NOX2 deficiency perturbed the immune system upon aging. NOX2 KO mice demonstrated preferred development of CD11b+Gr-1+ myeloid cells with profound production of proinflammatory cytokines and augmented expression of IL-17 through the activation of STAT3 and RORγt in vivo. NOX2 deficiency increased differentiation of effector Th cells in vitro and decreased CD25+FoxP3+ Treg cells both in vitro and in vivo. Furthermore, adoptive transfer of NOX2-deficient CD4(+) T cells into RAG KO mice increased arthritic inflammation compared with WT cells. These results demonstrated that NOX2 deficiency affected the development of CD11b+ myeloid cells and Th17/Treg cells, and thus promoted inflammatory cytokine production and inflammatory arthritis development, strongly supporting a crucial role for ROS generation in the modulation of Th17/Treg cell development and its related inflammatory immune response upon aging.


Hepatic phenotypes of HNF1B gene mutations: a case of neonatal cholestasis requiring portoenterostomy and literature review.
Hepatocyte nuclear factor 1-β (HNF1B) defects cause renal cysts and diabetes syndrome (RCAD), or HNF1B-maturity-onset diabetes of the young. However, the hepatic phenotype of HNF1B variants is not well studied. We present a female neonate born small for her gestational age [birth weight 2360 g; -2.02 standard deviations (SD) and birth length 45 cm; -2.40 SD at the 38(th) gestational week]. She developed neonatal cholestasis due to biliary atresia and required surgical intervention (portoenterostomy) when 32-d old. Following the operation, icterus resolved, but laboratory signs of liver dysfunction persisted. She had hyperechogenic kidneys prenatally with bilateral renal cysts and pancreatic hypoplasia postnatally that led to the diagnosis of an HNF1B deletion. This represents the most severe hepatic phenotype of an HNF1B variant recognized thus far. A review of 12 published cases with hepatic phenotypes of HNF1B defects allowed us to distinguish three severity levels, ranging from neonatal cholestasis through adult-onset cholestasis to non-cholestatic liver impairment, all of these are associated with congenital renal cysts and mostly with diabetes later in life. We conclude that to detect HNF1B variants, neonates with cholestasis should be checked for the presence of renal cysts, with special focus on those who are born small for their gestational age. Additionally, patients with diabetes and renal cysts at any age who develop cholestasis and/or exocrine pancreatic insufficiency should be tested for HNF1B variants as the true etiological factor of all disease components. Further observations are needed to confirm the potential reversibility of cholestasis in infancy in HNF1B mutation/deletion carriers.


Chinese Immigrant Entrepreneurship in the United States: Temporal and Spatial Dimensions.
Focusing on transnational entrepreneurship and immigrant businesses in new destinations, this paper studies entrepreneurship of Chinese immigrants in the U.S. using data from three surveys. In the analysis of transnational connections, we focus on the temporal dimension that links pre-migration and post-migration business activities. Results from logistic models reveal that the prospect of being self-employed among Chinese immigrants is significantly enhanced if they are from households in China with business backgrounds. This finding highlights the fact that transnational entrepreneurship is embedded in the multi-stranded connections between the immigrant sending and receiving societies. In the second part of the paper, sequence analysis is used to describe and classify business trajectories in traditional and new immigrant destinations. The results establish that while it may take a longer time for immigrants to achieve business ownership in new destinations than in traditional destinations, new immigrant destinations increase the chance of business expansion from one business to multiple businesses. These findings indicate a transition in immigrant entrepreneurs' business models. Businesses in traditional destinations mainly follow a survival strategy, while those in new destinations are adopting models that are akin to mainstream business operations, which gives rise to more opportunities for socioeconomic mobility.


Imaging tumor angiogenesis with fluorescent proteins.
We have developed three unique mouse models to image angiogenesis with fluorescent proteins, which are described in this review. First, we have adapted the surgical orthotopic implantation (SOI) model to image angiogenesis of human tumors labeled with green fluorescent protein (GFP) transplanted in nude mice. The nonluminous induced capillaries are clearly visible by contrast against the very bright tumor fluorescence examined either intravitally or by whole-body imaging in real time. Intravital images of an SOI model of human pancreatic tumors expressing GFP visualized angiogenic capillaries at both primary and metastatic sites. Whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period in an SOI model of human breast cancer expressing GFP. Opening a reversible skin-flap in the light path markedly reduces signal attenuation, increasing detection sensitivity many-fold and enabling vessels to be externally visualized in GFP-expressing tumors growing on internal organs. The second model utilizes dual-color fluorescence imaging, effected by using red fluorescent protein (RFP)-expressing tumors growing in GFP-expressing transgenic mice that express GFP in all cells. This dual-color model visualizes with great clarity the details of the tumor-stroma interaction, especially tumor-induced angiogenesis. The GFP-expressing tumor vasculature, both nascent and mature, are readily distinguished interacting with the RFP-expressing tumor cells. Using a spectral imaging system based on liquid crystal tunable filters, we were able to separate individual spectral species on a pixel-by-pixel basis. Such techniques non-invasively visualized the presence of host GFP-expressing vessels within an RFP-labeled orthotopic human breast tumor by real-time whole-body imaging. The third model involves a transgenic mouse in which the regulatory elements of the stem cell marker nestin drive GFP. The nestin-GFP mouse expresses GFP in areas of the brain, hair follicle stem cells, and in a network of blood vessels in the skin interconnecting hair follicles. RFP-expressing tumors transplanted to nestin-GFP mice enable specific visualization of nascent vessels in skin-growing tumors such as melanoma. Thus, fluorescent proteins expressed in vivo offer very high resolution and sensitivity for real-time imaging of angiogenesis.


Cellular responses to hypoxia in the pulmonary circulation.
Hypoxia can be defined as a reduction in available oxygen, whether in a whole organism or in a tissue or cell. It is a real life cause of pulmonary hypertension in humans both in terms of patients with chronic hypoxic lung disease and people living at high altitude. The effect of hypoxia on the pulmonary vasculature can be described in two ways; Hypoxic pulmonary vasoconstriction (HPV) (resulting from smooth muscle cell contraction) and pulmonary vascular remodelling (PVR) (resulting from pulmonary vascular cell proliferation). The pulmonary artery is made up of three resident cell types, the endothelial (intima), smooth muscle (media) and fibroblast (adventitia) cells. This review will examine the effects of hypoxia on the cells of the pulmonary vasculature and give an insight into the possible underlying mechanisms.


Natural variability of respiratory-syncytial viruses, their interference and reproduction characteristics.
Over a 5-year observation period in a children-institution community, the severity of respiratory-syncytial (RS) virus infection varied within a broad range depending on biological properties of the circulating virus population. The type of virus population was characterized by a complex of biological features, namely, the extent of their reproduction and interference in tissue cultures of different origin at optimal incubation temperature. The dynamics of genetic alterations in respect to these properties within 1976-1980 was established. Moreover, the seasonal variability of the strains with respect to these properties has been determined.


Estradiol regulates the slow Ca2+-activated K+ current in hippocampal pyramidal neurons.
The slow Ca2+-activated K+ current (sIAHP) was recorded in CA1 pyramidal neurons in hippocampal slices obtained from ovariectomized (OVX) or sham OVX (control) female rats. The sIAHP was significantly larger in cells from OVX rats than in cells from control rats. Superfusion with 5-100 nm 17beta-estradiol (E2) caused a progressive decrease in the sIAHP in cells from OVX rats but not in cells from control rats. In slices from OVX rats injected with 10 microg of E2 24 and 48 hr before they were killed, superfusion with E2 did not modify the sIAHP. In neurons from OVX rats, but not in neurons from control rats, E2 significantly increased both the number of action potentials and the burst duration generated by depolarizing pulses. The inactive isomer 17alpha-estradiol had no effect. The impermeant protein conjugate E2--BSA was as effective as free E2 at decreasing the sIAHP. Ca2+ spikes were also depressed by E2 in neurons from OVX rats, but not in control rats. A decrease in the intracellular Ca2+ signal, correlating with the inhibition of the Ca2+ spike and sIAHP produced by E2, was observed only in neurons from OVX rats. Our results indicate that ovariectomy increases the sIAHP and depresses excitability, whereas bath application or priming with E2 decreases the sIAHP, thus promoting excitability. These effects of E2 on the sIAHP and excitability, which are stereospecific and presumably mediated by membrane-bound receptors, could contribute to the hormonal regulation of synaptic plasticity and epileptiform activity as well as to learning and cognitive abilities dependent on the function of hippocampal neural circuits.


Microvascular free flap based on the medial femoral main cutaneous artery.
This article introduces a free flap based on the medial femoral main cutaneous artery. Flap transfer was carried out in two cases with complete postoperative success. An anatomical description of the midportion of the medial femoral region, based on magnification and binocular dissection microscope studies is provided, and the operative procedure is described.


High Leptospira Diversity in Animals and Humans Complicates the Search for Common Reservoirs of Human Disease in Rural Ecuador.
Leptospirosis is a zoonotic disease responsible for high morbidity around the world, especially in tropical and low income countries. Rats are thought to be the main vector of human leptospirosis in urban settings. However, differences between urban and low-income rural communities provide additional insights into the epidemiology of the disease. Our study was conducted in two low-income rural communities near the coast of Ecuador. We detected and characterized infectious leptospira DNA in a wide variety of samples using new real time quantitative PCR assays and amplicon sequencing. We detected infectious leptospira in a high percentage of febrile patients (14.7%). In contrast to previous studies on leptospirosis risk factors, higher positivity was not found in rats (3.0%) but rather in cows (35.8%) and pigs (21.1%). Six leptospira species were identified (L. borgpetersenii, L kirschnerii, L santarosai, L. interrogans, L noguchii, and an intermediate species within the L. licerasiae and L. wolffii clade) and no significant differences in the species of leptospira present in each animal species was detected (χ2 = 9.89, adj.p-value = 0.27). A large portion of the world's human population lives in low-income, rural communities, however, there is limited information about leptospirosis transmission dynamics in these settings. In these areas, exposure to peridomestic livestock is particularly common and high prevalence of infectious leptospira in cows and pigs suggest that they may be the most important reservoir for human transmission. Genotyping clinical samples show that multiple species of leptospira are involved in human disease. As these genotypes were also detected in samples from a variety of animals, genotype data must be used in conjunction with epidemiological data to provide evidence of transmission and the importance of different potential leptospirosis reservoirs.


Surfactants from itaconic acid: Toxicity to HaCaT keratinocytes in vitro, micellar solubilization, and skin permeation enhancement of hydrocortisone.
One of the most widely used approaches for improving drug permeation across the stratum corneum barrier of the skin is the use of chemical penetration enhancers, such as surfactants. In this study, two anionic surfactants, named C12-OPK and C18-OPK, were synthesized via condensation of itaconic acid and fatty amines, with C12 and C18 alkyl chains, respectively. Assessment of impacts on HaCaT keratinocyte cell viability was used as indicator of their potential to cause skin irritation 24h post exposure (Alamar Blue assay). The LC50 values of C12-OPK and C18-OPK (144 and 85mg/L, respectively) were lower than LC50 values of the most used commercial surfactants (e.g. SDS). The effect of different surfactant concentrations (up to ten times the critical micellar concentration, CMC) on hydrocortisone (HC) solubility and permeation through porcine skin was also evaluated. Results showed that drug solubility increased linearly with increasing concentrations of both surfactants, as a consequence of the association between drug and micelles. In vitro permeation results showed that the permeability coefficient increased at surfactant concentrations lower than the CMC. In particular, a higher enhancement effect on drug permeation was obtained with C18-OPK, due to its hydrophobic properties that ensured a more effective HC permeation in comparison to C12-OPK.


Conjunctival Melanocytic Lesions.
Conjunctival melanocytic lesions consist of a variety of neoplastic and nonneoplastic conditions. These include benign processes such as primary intraepithelial hypermelanosis and melanocytic hyperplasia, secondary forms of intraepithelial hypermelanosis and melanocytic hyperplasia, melanocytic nevi, melanocytic proliferations with malignant potential, and melanoma. To provide a concise yet comprehensive resource regarding the histopathologic diagnosis of conjunctival melanocytic lesions. We aim to detail and clarify the numerous classification schemes that exist for junctional melanocytic proliferations of the conjunctiva (known as primary acquired melanosis or PAM; also termed conjunctival melanocytic intraepithelial neoplasia or C-MIN). Although not uniformly adopted, C-MIN is classified by using a numeric system based on a defined set of criteria. A less complex scheme (conjunctival melanocytic intraepithelial lesion or CMIL) has recently been proposed by the World Health Organization. Additionally, we aim to update the reader regarding molecular features and prognostic indicators. Peer-reviewed literature and archived cases for illustration. Accurate histologic classification is essential, as PAM/C-MIN/CMILs that have a significant potential to progress to invasive melanoma may be clinically indistinguishable from low-risk lesions. Conjunctival melanoma (CM) more closely resembles cutaneous melanoma in terms of its pathogenesis and molecular features, compared to melanoma arising at other mucosal sites or to uveal melanoma. Depth of invasion and ulceration status, among other factors, have emerged as important prognostic indicators in CM. Sentinel lymph node biopsy may provide further prognostic information. Lastly, integration of pathologic and clinical findings is essential at this anatomically sensitive location to determine appropriate clinical management.


Synaptic input and local output of dopaminergic neurons in grafts that functionally reinnervate the host neostriatum.
In adult rats with a unilateral 6-hydroxy-dopamine-induced lesion of the nigrostriatal dopamine pathway, grafts of embryonic ventral mesencephalon can establish extensive efferent connections with the previously denervated host neostriatum and can compensate for motor and sensorimotor asymmetries induced by the lesion. The object of this study was to examine the afferent synaptic inputs to grafted dopaminergic neurons, implanted into a cortical cavity overlying the previously denervated caudate-putamen, using electron microscopic immunocytochemistry. The dopaminergic neurons of the grafts in the same animals had previously been shown to re-innervate the host neostriatum, to form synaptic connections therein and to attenuate the lesion-induced motor asymmetry that occurred in response to amphetamine (Freund et al. 1985). In the light microscope, the grafts were found to contain numerous tyrosine hydroxylase-immunoreactive perikarya, dendrites, axons and axonal swellings which had distinct distributions. In addition axons and axonal swellings that were immunoreactive for either substance P or glutamate decarboxylase were present. Electron microscopic analysis of the boutons contacting tyrosine hydroxylase-immunoreactive neurons in the grafts revealed the presence of at least five distinct types of afferent synaptic boutons based on their immunochemistry, morphology, or types of membrane specialization. One type was itself immunoreactive for tyrosine hydroxylase; such synapses are extremely rare in the intact substantia nigra, none were found in the contralateral substantia nigrae or the substantia nigra of a control rat. Three of the remaining types had ultrastructural features that were similar to synaptic terminals that were immunoreactive for substance P or glutamate decarboxylase. These synapses were similar to the types of synapses found contacting dopaminergic neurons in the substantia nigra contralateral to the graft or the substantia nigra of a control rat. The results demonstrate that, in the absence of the normal extrinsic afferent inputs, the intracortical mesencephalic grafts have a well-developed local synaptic circuitry. It is suggested that local circuit regulation of dopaminergic neurons within the graft may, at least in part, be responsible for the maintenance of a normal or close to normal functional activity.


Age-related patterns in high-risk alcohol and cannabis use and their associations with positive and negative affect in young adulthood.
We examined age-varying associations between young adult simultaneous alcohol and marijuana/cannabis use (SAM) and heavy episodic drinking (HED) and positive and negative affect to inform harm reduction efforts. Young adults reporting past-year alcohol use (n = 556; ages 19-25) were recruited in a state where alcohol and nonmedical cannabis use was legal for those 21 +. Participants provided 24 repeated monthly assessments. Among those reporting past-month cannabis use on at least one survey, logistic time-varying effect models estimated (1) the age-varying prevalence of and associations between past-month SAM and HED and (2) age-varying unique associations of affect with SAM and HED. There was a positive age-varying association between HED and SAM over time that was highest at age 19 (OR = 7.56), decreased until age 20.7 (OR = 3.39), increased until age 23.0 (OR = 4.85), and decreased until the association became non-significant by age 25. Negative affect was positively associated with SAM from ages 20.7 to 23.0, peaking at age 21.8 (OR = 1.36). Positive affect was positively associated with HED from ages 19.4 to 20.4 (peak OR = 1.25) and ages 22.5 to 24.5 (peak OR = 1.38). In contrast, positive affect was not uniquely associated with SAM nor negative affect with HED across ages 19-25. While HED and SAM were positively associated throughout young adulthood and interventions could target them in tandem, their associations with affect suggest differential etiologic processes. Preventive intervention and harm reduction efforts should attend to psychological context in which these behaviors occur.


Using spectrophotometric titrations to characterize humic acid reactivity at environmental concentrations.
Potentiometric titration is a common method to characterize dissolved organic matter (DOM) reactivity. Because of the sensitivity of pH electrodes, it is necessary to work with very high DOM (>1 g/L) concentrations that are unrealistic compared to those found in natural waters (0.1 to 100 mg/L). To obtain proton binding data for concentrations closer to environmental values, spectroscopic titration methodology is a viable alternative to traditional potentiometric titrations. Spectrophotometric titrations and UV-visible spectra of a diluted solution of purified Aldrich humic acid (5 mgDOC/L) are used to estimate changes in proton binding moieties as function of pH and ionic strength after calculation of differential absorbance spectra variations. After electrostatic correction of spectrophotometric data, there is a linear operational correlation between spectrophotometric and potentiometric data which can be used as a transfer function between the two properties. Spectrophotometric titrations are then used to determine the changes of humic acid protonation after adsorption onto alpha-alumina.


[Prognosis after acute coronary syndrome. Lack of difference according to the sex].
Acute coronary syndromes (ACS) represent a continuum from unstable angina to non-ST-elevation myocardial infarction. ACS is the preferred diagnostic term for non transmural myocardial infarction. The different forms of ACS share a common anatomic substrate consisting of atherosclerotic plaque rupture or erosion, with variable degrees of thrombus formation and compromised blood flow to viable myocardium. Patients with ACS have a heterogeneous profile of short- and mid-term adverse outcomes and require a tailored approach. Recent major advances in the management of ACS have emphasized the importance of earlier identification of higher-risk patients, whose outcome might be improved by aggressive revascularization. However, during the past several years, numerous studies have shown a significant difference in the prognosis of women and men with ACS. Some reports have concluded that women have a worse prognosis than men after thrombolytic therapy and/or coronary angioplasty with stenting for myocardial reperfusion. The aim of our retrospective study was to determine sex differences in outcomes after early percutaneous intervention (PCI) in high-risk patients with acute coronary syndromes (ACS). A total of 694 consecutive patients (151 women with 233 treated lesions and 543 men with 850 treated lesions) were included. Enrollments were limited to ACS patients judged to be at high risk [unstable angina/non ST-elevation myocardial infarction with recurrent ischemia/dynamic ST segment changes (53.6% vs 52.4%) or post-infarction unstable angina (46.4% vs 47.6%)] who underwent PCI within 24 hours of admission if the coronary anatomy was deemed suitable. The two groups were well matched for clinical and lesion characteristics, except that the women, as in other studies, were older (67.9 +/- 11.3 vs 62.3 +/- 12.3 years) and had a higher prevalence of hypertension. All the lesions were treated by stent implantation, and glycoprotein IIb/IIIa inhibitors were used similarly in the two sexes (27.1% of women, 30.5% of men). The success rates were similar (94% and 93.7%), with a similar incidence of in-hospital MACE (4% vs 3.8%, p = 0.56). After a mean follow-up of 564 +/- 294 days, the two groups had similar rates of mortality (2% vs 3.2%), myocardial infarction (6.7% vs 7.1%) and repeated revascularization (7.2% vs 8.3%). The respective event-free survival rates were 88 +/- 60.3% and 83 +/- 0.3% at 1 year and 87% vs 78 +/- 0.2% at 2 years (p = 0.58). Previous studies have shown increased morbidity and mortality associated with recurrent ischemia and myocardial infarction in women after acute revascularization for ACS. Our study confirms that aggressive revascularization offers a comparable survival benefit in the two sexes. Women with ACS who are at high risk derive the same benefit as men from early angioplasty and coronary stenting of the culprit lesion, with satisfactory in-hospital and mid-term outcomes.


Neuro-optic cell apoptosis and microangiopathy in KKAY mouse retina.
Diabetic retinopathy is one of the common complications of diabetes and is the leading cause for patients' visual dysfunction and sight loss. However, the mechanism of diabetic retinopathy is not clearly defined. The present study was undertaken to investigate neuroretinal apoptosis in different stages in a mouse model for type 2 diabetes mellitus and the mechanism of diabetic retinopathy. KKAY mouse with genetic diabetes, an animal model for type 2 diabetes, was used in this study. Mice were divided into a control group and a diabetic group. The mice in both groups were sacrificed at one month and three months, and the mouse eyeballs were used for making retinal histologic sections. We showed in this study that the apoptotic cell numbers for retinal neural cells in the ganglion cell layer were significantly greater in the diabetic group than in the control group (p<0.01), as determined by the TUNEL assay. In addition, many more apoptotic retinal neuronal cells were found in the retinal ganglion cell layer and medial part of the inner nuclear layer in the diabetic group when the mice were sacrificed at three months as compared to those sacrificed at one month (p<0.01). We also studied the ultrastructure of the retinal nerve cells and microvesseles by electron microscopy and demonstrated that the ultrastructure changes for retinal neural cells and retinal microangiopathy were observed in, as early as, the early stage of diabetes. These findings indicate that: i). retinal neuropathy and microangiopathy occur in the early stage of diabetes, ii). apoptosis may be an important mechanism through which retinal neurodegeneration is developed, and iii). both retinal neurodegeneration and retinal microangiopathy should be considered as the diabetic retinopathy.


Biological sensitivity to context: Cortisol awakening response moderates the effects of neighbourhood density on the development of adolescent externalizing problem behaviours.
This four-year longitudinal study attempted to test person-environment interaction theory and biological sensitivity theory by assessing whether individuals' biological stress activity CARAUCg (Cortisol Awakening Response Area Under the Curve with respect to ground) moderates the effects of neighbourhood density on the development of adolescent externalizing problem behaviours. Participants were 358 Dutch adolescents with a mean age of 15 years at the first measurement. Our analyses showed that CARAUCg moderated the effects of neighbourhood density on the level of parent-reported delinquency and aggression and adolescent self-reported delinquency. More specifically, for adolescents with high CARAUCg, higher neighbourhood density significantly predicted higher levels of parent-reported and adolescent self-reported delinquency and aggression, whereas the association was reversed or non-significant for adolescents with low CARAUCg. Our findings suggest that adolescents with different levels of CARAUCg respond differentially to the density of the neighbourhood they live in, supporting for person-environment interaction perspectives and biological sensitivity theory.


Analysis of a noncanonical poly(A) site reveals a tripartite mechanism for vertebrate poly(A) site recognition.
At least half of all human pre-mRNAs are subject to alternative 3' processing that may modulate both the coding capacity of the message and the array of post-transcriptional regulatory elements embedded within the 3' UTR. Vertebrate poly(A) site selection appears to rely primarily on the binding of CPSF to an A(A/U)UAAA hexamer upstream of the cleavage site and CstF to a downstream GU-rich element. At least one-quarter of all human poly(A) sites, however, lack the A(A/U)UAAA motif. We report that sequence-specific RNA binding of the human 3' processing factor CFI(m) can function as a primary determinant of poly(A) site recognition in the absence of the A(A/U)UAAA motif. CFI(m) is sufficient to direct sequence-specific, A(A/U)UAAA-independent poly(A) addition in vitro through the recruitment of the CPSF subunit hFip1 and poly(A) polymerase to the RNA substrate. ChIP analysis indicates that CFI(m) is recruited to the transcription unit, along with CPSF and CstF, during the initial stages of transcription, supporting a direct role for CFI(m) in poly(A) site recognition. The recognition of three distinct sequence elements by CFI(m), CPSF, and CstF suggests that vertebrate poly(A) site definition is mechanistically more similar to that of yeast and plants than anticipated.


Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis.
Mosquito salivary proteins are involved in several biological processes that facilitate their blood feeding and have also been reported to elicit an IgG response in vertebrates. A growing number of studies have focused on this immunological response for its potential use as a biological marker of exposure to arthropod bites. As mosquito saliva collection is extremely laborious and inefficient, most research groups prefer to work on mosquito salivary glands (SGs). Thus, SG protein integrity is a critical factor in obtaining meaningful data from immunological and biochemical analysis. Current methodologies rely on an immediate freezing of SGs after their collection. However, the maintenance of samples in a frozen environment can be hard to achieve in field conditions. In this study, SG proteins from two mosquito species (Aedes aegypti and Anopheles gambiae s.s.) stored in different media for 5 days at either +4°C or room temperature (RT) were evaluated at the quantitative (i.e., ELISA) and qualitative (i.e., SDS-PAGE and immunoblotting) levels. Our results indicated that PBS medium supplemented with an anti-protease cocktail seems to be the best buffer to preserve SG antigens for 5 days at +4°C for ELISA analysis. Conversely, cell-lysis buffer (Urea-Thiourea-CHAPS-Tris) was best at preventing protein degradation both at +4°C and RT for further qualitative analysis. These convenient storage methods provide an alternative to freezing and are expected to be applicable to other biological samples collected in the field.


Course and predictors of posttraumatic stress and depression longitudinal symptom profiles in refugees: A latent transition model.
Exposure to potentially traumatic events and post-migration living difficulties (PMLDs) may explain the high rates of posttraumatic stress disorder (PTSD) and depression in resettled refugees. Latent class analyses (LCAs) in refugees have identified subgroups that differ in symptom profiles of PTSD and comorbid symptoms. However, knowledge on longitudinal symptom profiles in refugees is sparse. Examining longitudinal PTSD and depression symptom profiles could provide information on risk factors underlying worsening of symptoms post-resettlement. Self-rated PTSD (Posttraumatic Diagnostic Scale) and depression (Patient Health Questionnaire-9) symptoms were assessed among 613 refugees who had resettled in Australia up to two years previously (W1) and at 6 months follow-up (W2). PTSD and depression symptom profiles were identified using LCAs for W1 and W2 separately. Latent transition analysis was used to examine (predictors of) changes in symptom profiles, including gender, age, trauma exposure, and PMLDs. Four classes were identified that were consistent across timepoints: a No symptoms (W1 61%; W2 68%), Low PTSD/Moderate depression (W1 16%; W2 10%), Moderate PTSD/depression (W1 16%; W2 14%), and High symptoms class (W1 7%; W2 7%). Higher levels of problems with PMLDs, including being discrimination and family separation, predicted movements out of the No symptom class at W1 to classes with psychopathology at W2. To conclude, most participants did not develop PTSD or depression symptoms. The risk of developing these symptoms seems higher when problems with interpersonal PMLDs increased, pointing to the need for considering these stressors when addressing the mental health needs in this population.


Role of myokines in the maintenance of whole-body metabolic homeostasis.
Obesity is reaching epidemic proportions in developed countries and is on the rise in developing countries. Obesity-related changes in lipid and glucose metabolism predispose to the development of metabolic syndrome and type 2 diabetes. Skeletal muscle constitutes about 40 percent of total body weight and is unique compared to other muscle types since it is one of the most important organs for insulin-dependent glucose metabolism in humans. Abnormalities in skeletal muscle lipid and glucose metabolism as well as abnormal accumulation of intramyocellular lipids could predispose for the development of type 2 diabetes. Skeletal muscle synthesizes and secretes factors with autocrine/paracrine/endocrine functions that can regulate skeletal muscle metabolism as well as affect other organs. These factors secreted by skeletal muscle are called myokines. Secretion and action of myokines is regulated by physiological conditions. Some myokines have positive effect on metabolism, improving functions of multiple organs. Yet, other myokines are released under pathological conditions and might exacerbate abnormal metabolic functions. Expression and/or secretion of a number of myokines are regulated by exercise and therefore might mediate positive effects of physical activity on whole-body metabolism. In the current review we summarized current knowledge on some of the myokines with important physiological functions in lipid and glucose metabolism. A better understanding of the effects of myokines on whole-body metabolism can aid in development of the future pharmacologic therapies for counteracting the current worldwide obesity epidemic and obesity-mediated abnormalities.


Can autism be triggered by acetaminophen activation of the endocannabinoid system?
Acetaminophen use in children has been associated with increased autism risk. Recent evidence suggests that acetaminophen's analgesic actions result from activation of the endocannabinoid system, and activation of this system can have neuromodulatory consequences during development. This investigation was performed to determine if there is evidence to support the hypothesis that acetaminophen use can trigger autism by activation of the endocannabinoid system.


Prevalence of overt dilated cardiomyopathy in two regions of England.
The prevalence of dilated cardiomyopathy was assessed in two regions in England from 420 replies to 771 questionnaires sent to general practitioners. Overall point prevalence was 8.317 per 100 000 population in areas covering a total sample of 913 836 inhabitants. There was a statistically significant regional variation between East Anglia (Norfolk, Suffolk, and Cambridgeshire) and Essex, Hertfordshire, and Bedfordshire. Though the natural history of dilated cardiomyopathy remains to be defined, this study has provided an assessment of point prevalence of overt cases of this condition.


Situating the Pandemic in the Life-Course: Older Adults' Perspectives on the Challenges and Positives of the COVID-19 Pandemic.
Older adults experienced the highest rates of infections and deaths and significant social isolation during the COVID-19 pandemic. While these negative impacts are important to address, the positive outcomes among older adults during the pandemic are equally important. A survey was distributed to adults aged 65 or older living in St. Louis, Missouri from August 2020 through March 2021 to characterize the challenges and positives they found during the pandemic. A total of 103 older adults participated. Compared to previous times in their lives, a majority of participants felt the pandemic had been more disruptive (62%) and more confusing (78%) and a majority were more worried (58%) and more afraid of dying (53%) during the pandemic. The most common positives were increased emotional well-being (23% of responses) and stronger personal connections (20% of responses). This ability to identify positive outcomes of the pandemic demonstrates the resilience of older adults and counters the ageist narrative that dominated the dialogue early in the pandemic, which depicted older adults as weak, vulnerable, and dispensable. As the pandemic evolves, nurturing and leveraging these positives will be key to combatting the cycle of fear and restrictions that the pandemic could bring.


Nose-to-Brain Delivery of Peptide Drugs Enhanced by Coadministration of Cell-penetrating Peptides: Therapeutic Potential for Dementia.
Recent reports suggest that peptide drugs such as insulin have the potential to serve as therapeutics in neurodegenerative diseases such as Alzheimer's disease. However, the transport of these drugs to the therapeutic target, the brain, is significantly hindered by the blood-brain barrier (BBB). Intranasal administration appears to be an ideal solution for drug delivery to the brain, bypassing the BBB, however the entry of peptide drugs into neuronal and epithelial cells in the olfactory mucosa remains low. In this study, we therefore examined whether intranasal coadministration of cell-penetrating peptides (CPPs) could improve nose-to-brain drug transport. In both mice and rats, we found that direct transport of insulin into the brain was significantly facilitated when coadministered with amphipathic CPP penetratin, and eventually insulin reached the deeper regions of the brain such as the hippocampus. In the mouse line senescence-accelerated mouse prone-8 (SAMP8), spatial learning tests demonstrated that long-term intranasal coadministration of insulin with penetratin improved mild memory loss in the early stages of dementia. In contrast, the severe cognitive dysfunction in the aged SAMP8 mice was preserved despite intranasal coadministration of insulin with penetratin. The immunohistological examination of the hippocampus suggested that enhanced nose-to-brain delivery of insulin had a partial neuroprotective effect but unexpectedly increased amyloid β plaque deposition. In conclusion, intranasal coadministration of insulin with CPPs has the potential to serve as a therapeutic for mild cognitive dysfunction. To identify suitable pharmacotherapy for dementia with severe pathology, further studies of nose-to-brain delivery of molecularly appropriate biopharmaceuticals are necessary.


Hepatoprotective Effect of Vitamin B12 in Acetaminophen Induce Hepatotoxicity in Male Rats.
Acetaminophen is a pharmaceutical synthesized non-opioid analgesic that belongs to the "aniline analgesics" class of medicine. Because it lacks a significant anti-inflammatory effect, it is not classified as a non-steroidal anti-inflammatory therapeutic medication (NSAID). As an over-the-counter pain reliever and antipyretic, Acetaminophen is the active metabolite of phenacetin and acetanilide, but it is less toxic than either precursor. According to some medical studies, Acetaminophen toxicity can be treated with vitamin B12. Acetaminophen-poisoned Male Wister rats were the subject model of the current study, which examines the effects of vitamin B12 on their hepatic health. There were three groups of animals: Acetaminophen treated animals (750 ml/kg), vitamin B12-treated animals (0.63 g/kg), and a control group that received distilled water (750 ml/kg). All animals were given oral medication for seven days. On the seventh day, the animal was sacrificed. Plasma levels of Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Glutathione (GSH), total antioxidant capacity (TAC), Caspase3, Malondialdehyde (MDA), Interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-) were measured in the cardiac blood samples. Vitamin B12 lowers liver enzyme levels in the blood, increases overall antioxidant levels, and compensates for tissue glutathione deficiency while lowering serum elevations. TNF-α and interleukin-6 levels are also reduced by caspase3. Acetaminophen-induced hepatic necrosis and inflammatory cell infiltration were both considerably reduced by vitamin B12 supplementation. According to this study, vitamin B12 was found to have a protective effect against acetaminophen-induced hepatotoxicity.


Effects of simulated cochlear-implant processing on speech reception in fluctuating maskers.
This study investigated the effects of simulated cochlear-implant processing on speech reception in a variety of complex masking situations. Speech recognition was measured as a function of target-to-masker ratio, processing condition (4, 8, 24 channels, and unprocessed) and masker type (speech-shaped noise, amplitude-modulated speech-shaped noise, single male talker, and single female talker). The results showed that simulated implant processing was more detrimental to speech reception in fluctuating interference than in steady-state noise. Performance in the 24-channel processing condition was substantially poorer than in the unprocessed condition, despite the comparable representation of the spectral envelope. The detrimental effects of simulated implant processing in fluctuating maskers, even with large numbers of channels, may be due to the reduction in the pitch cues used in sound source segregation, which are normally carried by the peripherally resolved low-frequency harmonics and the temporal fine structure. The results suggest that using steady-state noise to test speech intelligibility may underestimate the difficulties experienced by cochlear-implant users in fluctuating acoustic backgrounds.


Anti-alpha-fodrin autoantibodies in Moyamoya disease.
Moyamoya disease (MMD) is a rare entity that results in progressive occlusion of the arteries of the circle of Willis, but the pathogenesis of MMD is unknown. MMD sera (n=32) were tested for anti-endothelial cell antibodies by enzyme-linked immunoassays and flow cytometric analysis. Apoptosis was induced in human umbilical vein endothelial cells by tumor necrosis factor-alpha. We found that a high proportion of MMD sera had anti-endothelial cell antibodies with apoptotic stimuli. Prominent reactivities of MMD sera (72%) with recombinant human alpha-fodrin were observed. Our study demonstrates that MMD sera contain a high incidence of anti-alpha-fodrin autoantibodies, providing new insight into the mechanisms of occlusion of MMD arteries.


Repeatability of a novel technique for in vivo measurement of three-dimensional patellar tracking using magnetic resonance imaging.
To determine the repeatability of a novel noninvasive MRI-based technique for measuring patellofemoral kinematics in vivo. The patellar kinematics measurement method relies on registering bone models (with associated coordinate systems) developed from a high resolution MRI scan to loaded bone positions derived from fast, low resolution MRI scans. The intrasubject variability, high resolution to low resolution registration error, and interexperimenter repeatability were quantified in experiments on three healthy subjects. The intrasubject variability and registration error were within range of the accuracy of our procedure; specifically, less than or equal to 1.40 degrees for orientation and 0.81 mm for translation. The interexperimenter repeatability was less than or equal to 1.28 degrees for orientation, with the exception of patellar spin, and 0.68 mm for translation. Our novel measurement technique can measure three-dimensional patellar tracking noninvasively during loaded flexion in a repeatable manner. Our results compare well to another noninvasive tracking protocol, fast phase-contrast MRI, which has a reported subject interexam variability of 2.4 degrees or less for patellar orientation. A particular strength of our method is that axes and high-resolution bone models need only be determined once for intrasubject comparisons. The method is sufficiently accurate and repeatable to detect clinically significant changes in patellofemoral kinematics.


Murine noroviruses comprising a single genogroup exhibit biological diversity despite limited sequence divergence.
Viruses within the genus Norovirus of the family Caliciviridae are the major cause of acute, nonbacterial gastroenteritis worldwide. Human noroviruses are genetically diverse, with up to 57% divergence in capsid protein sequences, and comprise three genogroups. The significance of such genetic diversity is not yet understood. The discovery of murine norovirus (MNV) and its ability to productively infect cultured murine macrophages and dendritic cells has provided an opportunity to determine the functional consequences of norovirus diversity in vitro and in vivo. Therefore, we compared the full-length genomes of 21 new MNV isolates with five previously sequenced MNV genomes and demonstrated a conserved genomic organization consisting of four open reading frames (ORFs) and a previously unknown region of nucleotide conservation in ORF2. A phylogenetic analysis of all 26 MNV genomes revealed 15 distinct MNV strains, with up to 13% divergence at the nucleotide level, that comprise a single genotype and genogroup. Evidence for recombination within ORF2 in several MNV genomes was detected by multiple methods. Serological analyses comparing neutralizing antibody responses between highly divergent strains suggested that the MNV genogroup comprises a single serotype. Within this single genogroup, MNV strains exhibited considerable biological diversity in their ability to grow in culture and to infect and/or persist in wild-type mice. The isolation and characterization of multiple MNV strains illustrate how genetic analysis may underestimate the biological diversity of noroviruses and provide a molecular map for future studies of MNV biology.


Assessment of the effects of sugar cane plantation burning on daily counts of inhalation therapy.
This study was designed to evaluate the association between sugar cane plantation burning and hospital visits in Araraquara in the state of São Paulo, Brazil. From June 1 to August 31, 1995, the daily number of visits of patients who needed inhalation therapy in one of the main hospitals of the city was recorded and used as health impairment estimation. Sedimentation of particle mass (the amount of particles deposited on four containers filled with water) was measured daily. The association between the weight of the sediment and the number of visits was evaluated by means of Poisson regression models controlled for seasonality, temperature, day of the week, and rain. We found a significant and dose-dependent relationship between the number of visits and the amount of sediment. The relative risk of visit associated with an increase of 10 mg in the sediment weight was 1.09 (1-1.19), and the relative risk of an inhalation therapy was 1.20 (1.03-1.39) on the most polluted days (fourth quartile of sediment mass). These results indicate that sugar cane burning may cause deleterious health effects in the exposed population.


Early environment and neurobehavioral development predict adult temperament clusters.
Investigation of the environmental influences on human behavioral phenotypes is important for our understanding of the causation of psychiatric disorders. However, there are complexities associated with the assessment of environmental influences on behavior. We conducted a series of analyses using a prospective, longitudinal study of a nationally representative birth cohort from Finland (the Northern Finland 1966 Birth Cohort). Participants included a total of 3,761 male and female cohort members who were living in Finland at the age of 16 years and who had complete temperament scores. Our initial analyses (Wessman et al., in press) provide evidence in support of four stable and robust temperament clusters. Using these temperament clusters, as well as independent temperament dimensions for comparison, we conducted a data-driven analysis to assess the influence of a broad set of life course measures, assessed pre-natally, in infancy, and during adolescence, on adult temperament. Measures of early environment, neurobehavioral development, and adolescent behavior significantly predict adult temperament, classified by both cluster membership and temperament dimensions. Specifically, our results suggest that a relatively consistent set of life course measures are associated with adult temperament profiles, including maternal education, characteristics of the family's location and residence, adolescent academic performance, and adolescent smoking. Our finding that a consistent set of life course measures predict temperament clusters indicate that these clusters represent distinct developmental temperament trajectories and that information about a subset of life course measures has implications for adult health outcomes.


Evaluating how variants of floristic quality assessment indicate wetland condition.
Biological indicators are useful tools for the assessment of ecosystem condition. Multi-metric and multi-taxa indicators may respond to a broader range of disturbances than simpler indicators, but their complexity can make them difficult to interpret, which is critical to indicator utility for ecosystem management. Floristic Quality Assessment (FQA) is an example of a biological assessment approach that has been widely tested for indicating freshwater wetland condition, but less attention has been given to clarifying the factors controlling its response. FQA quantifies the aggregate of vascular plant species tolerance to habitat degradation (conservatism), and model variants have incorporated species richness, abundance, and indigenity (native or non-native). To assess bias, we tested FQA variants in open-canopy freshwater wetlands against three independent reference measures, using practical vegetation sampling methods. FQA variants incorporating species richness did not correlate with our reference measures and were influenced by wetland size and hydrogeomorphic class. In contrast, FQA variants lacking measures of species richness responded linearly to reference measures quantifying individual and aggregate stresses, suggesting a broad response to cumulative degradation. FQA variants incorporating non-native species, and a variant additionally incorporating relative species abundance, improved performance over using only native species. We relate our empirical findings to ecological theory to clarify the functional properties and implications of the FQA variants. Our analysis indicates that (1) aggregate conservatism reliably declines with increased disturbance; (2) species richness has varying relationships with disturbance and increases with site area, confounding FQA response; and (3) non-native species signal human disturbance. We propose that incorporating species abundance can improve FQA site-level relevance with little extra sampling effort. Using our practical sampling methods, an FQA variant ignoring species richness and incorporating non-native species and relative species abundance can be logistically efficient, easily understood, and effective for wetland assessment.


Clinically manifest thromboembolic complications of femoral vein catheterization for continuous renal replacement therapy.
The safety of femoral vein (FV) catheterization for continuous renal replacement therapy is uncertain. We sought to determine the incidence of clinically manifest venous thromboembolism (VTE) in such patients. We retrospectively studied patients with femoral high flow catheters (≥ 13F) (December 2005 to February 2011). Discharge diagnostic codes were independently screened for VTE. The incidence of VTE was also independently similarly assessed in a control cohort of patients ventilated for more than 2 days (January 2011 to December 2011) in the same intensive care unit (ICU). We studied 380 patients. Their mean age was 61 years, and 59% were male. The mean Acute Physiology and Chronic Health Evaluation III score was 84; average duration of continuous renal replacement therapy was 74 hours, and 232 patients (61%) survived to hospital discharge with an average length of hospital stay of 22 days. Only 5 patients (1.3%) had clinically manifest VTE after FV catheterization. In the control cohort of 514 ICU patients, the incidence of VTE was 4.4% (P < .05 compared with FV group). The incidence of clinically manifest VTE after FV catheterization with high flow catheters is low and lower to that seen in general ICU patients.


Human sex hormone-binding globulin variants associated with hyperandrogenism and ovarian dysfunction.
The access of testosterone and estradiol to target tissues is regulated by sex hormone-binding globulin (SHBG) in human blood. Serum SHBG levels are low in patients with hyperandrogenism, especially in association with polycystic ovarian syndrome (PCOS) and in individuals at risk for diabetes and heart disease. Here, we identify SHBG coding region variations from a compound heterozygous patient who presented with severe hyperandrogenism during pregnancy. Serum SHBG levels in this patient measured 2 years after her pregnancy were exceptionally low, and her non-protein-bound testosterone concentrations greatly exceeded the normal reference range. A single-nucleotide polymorphism within the proband's maternally derived SHBG allele encodes a missense mutation, P156L, which allows for normal steroid ligand binding but causes abnormal glycosylation and inefficient secretion of SHBG. This polymorphism was identified in four other patients with either PCOS, ioiopathic hirsutism, or ovarian failure. The proband's paternal SHBG allele carries a single-nucleotide deletion within exon 8, producing a reading-frame shift within the codon for E326 and a premature termination codon. CHO cells transfected with a SHBG cDNA carrying this mutation fail to secrete the predicted truncated form of SHBG. To our knowledge, these are the first examples of human SHBG variants linked to hyperandrogenism and ovarian dysfunction.


Quality of delivered care for people with type 2 diabetes: a new patient-centred model.
The quality of care from the perspective of people with Type 2 diabetes using a new model (CQMH) including three dimensions of quality in health care (Technical, Service and Customer Quality) was assessed. A cross-sectional survey with a sample of 577 people with Type 2 diabetes was conducted. Measures were self-reported adherence to national guidelines for technical quality, the Netherlands Institute for Health Services Research questionnaire for service quality and the short form of the Patient Activation Measure for Customer Quality. There was a significant gap in technical quality between what diabetes care the patients reported receiving and what was recommended in the guideline, particularly for management and lifestyle aspects. For service quality, the lowest scores were for choice of care provider and accessibility of care. The mean Customer Quality score was 64.5 (meaning higher score indicating better quality). A positive relationship was demonstrated between higher technical, service and customer quality scores, and better diabetes control status as well as maintaining continuity of care. The average Quality Index was 70.0 of a 0-100 scale. Customer Quality appears to be a useful third dimension in conceptualising quality in health care, particularly in the context of chronic disease, where good self-management can improve the outcomes of care. A high proportion of Queensland adults with Type 2 diabetes reported receiving suboptimal care in the majority aspects of provided care services as reflected in the overall Quality Index score indicating substantial room for quality improvement.


Differential diagnosis between dilated cardiomyopathy and ischemic cardiomyopathy based on variational mode decomposition and high order spectra analysis.
The clinical manifestations of ischemic cardiomyopathy (ICM) bear resemblance to dilated cardiomyopathy (DCM). The definitive diagnosis of DCM necessitates the identification of invasive, costly, and contraindicated coronary angiography. Many diagnostic studies of cardiovascular disease have tried modal decomposition based on electrocardiogram (ECG) signals. However, these studies ignored the connection between modes and other fields, thus limiting the interpretability of modes to ECG signals and the classification performance of models. This study proposes a classification algorithm based on variational mode decomposition (VMD) and high order spectra, which decomposes the preprocessed ECG signal and extracts its first five modes obtained through VMD. After that, these modes are estimated for their corresponding bispectrums, and the feature vector is composed of fifteen features including bispectral, frequency, and nonlinear features based on this. Finally, a dataset containing 75 subjects (38 DCM, 37 ICM) is classified and compared using random forest (RF), decision tree, support vector machine, and K-nearest neighbor. The results show that, in comparison to previous approaches, the technique proposed provides a better categorization for DCM and ICM of ECG signals, which delivers 98.21% classification accuracy, 98.22% sensitivity, and 98.19% specificity. And mode 3 always has the best performance among single mode. The proposed computerized framework significantly improves automatic diagnostic performance, which can help relieve the working pressure on doctors, possible economic burden and health threaten.


Determination of body fluids by the impedance technique.
For the purposes of determining body composition, the body can be viewed as the sum of two components, fat and fat-free mass, the latter including water, protein, and minerals. Current methods for determining the size of these various components of the body are either inexact, invasive, or dependent on expensive or slow instrumentation. Bioelectrical impedance, which is rapid, noninvasive, and relatively inexpensive is examined as an alternative method for measuring body composition. The potential for the impedance technique arises from the very different electrical properties of the body components. The theory of bioelectrical impedance analysis is briefly presented, and human applications are discussed. The results of studies validating the method are cited.


Analyzing for co-localization of proteins at a cell membrane.
Cell-to-cell communication is mediated by molecular interactions at the surface of the cell by soluble ligands released from distant cells or by cell surface molecules on adjacent cells. These interactions lead to activation of intracellular signaling pathways that subsequently can lead to activation of specific genes. This signal transduction process controls cellular activities as diverse as proliferation, differentiation and apoptosis, so we must understand the underlying molecular events in detail in order to understand broader questions related to development, uncontrolled growth in tumors, tissue regeneration and use of stem cells to name a few. Binding of a ligand in the extracellular space to a transmembrane receptor constitutes the first crucial step for activation of a signaling pathway within the cell. This binding can either lead to oligomerization of individual receptors, to reorganization of existing clusters of receptors or to changes in the protein conformations, which in turn results in recruitment of signaling molecules in the cytoplasm. While different membrane receptors activate different downstream signaling pathways, some receptors can activate more than one pathway and a particular pathway can be activated by different receptors. It appears that these processes are regulated either by agonists and antagonists in the extracellular medium, by receptor-receptor interactions in the membrane or by a number of signaling mediators in the cytoplasm of the cell. Our work has focused on understanding how the intermolecular interactions in the membrane can control the signal transduction process: Are there specialized structures on the surface that facilitate receptor-receptor interactions? Do the receptors exist as monomers or pre-existing complexes that enhance the probability of activation? Do different receptors associate in the same domains or are there distinct organizational principles for each receptor type. In order to address these questions, we seek to develop tools that allow us to examine intermolecular interactions and reactions directly on the cell surface, particularly on live cells in culture or in tissue. This review discusses some of the approaches that are currently available and highlights some of the key advantages and disadvantages they represent with particular focus on image cross correlation spectroscopy as a relatively new quantitative tool developed by us to address some of these issues.


Latent Rheumatic Heart Disease: Identifying the Children at Highest Risk of Unfavorable Outcome.
Screening echocardiography has emerged as a potentially powerful tool for early diagnosis of rheumatic heart disease (RHD). The utility of screening echocardiography hinges on the rate of RHD progression and the ability of penicillin prophylaxis to improve outcome. We report the longitudinal outcomes of a cohort of children with latent RHD and identify risk factors for unfavorable outcomes. This was a prospective natural history study conducted under the Ugandan RHD registry. Children with latent RHD and ≥1 year of follow-up were included. All echocardiograms were re-reviewed by experts (2012 World Heart Federation criteria) for inclusion and evidence of change. Bi- and multivariable logistic regression, Kaplan-Meier analysis, and Cox proportional hazards models, as well, were developed to search for risk factors for unfavorable outcome and compare progression-free survival between those treated and not treated with penicillin. Propensity and other matching methods with sensitivity analysis were implemented for the evaluation of the penicillin effect. Blinded review confirmed 227 cases of latent RHD: 164 borderline and 63 definite (42 mild, 21 moderate/severe). Median age at diagnosis was 12 years and median follow-up was 2.3 years (interquartile range, 2.0-2.9). Penicillin prophylaxis was prescribed in 49.3% with overall adherence of 84.7%. Of children with moderate-to-severe definite RHD, 47.6% had echocardiographic progression (including 2 deaths), and 9.5% had echocardiographic regression. Children with mild definite and borderline RHD showed 26% and 9.8% echocardiographic progression and 45.2% and 46.3% echocardiographic improvement, respectively. Of those with mild definite RHD or borderline RHD, more advanced disease category, younger age, and morphological mitral valve features were risk factors for an unfavorable outcome. Latent RHD is a heterogeneous diagnosis with variable disease outcomes. Children with moderate to severe latent RHD have poor outcomes. Children with both borderline and mild definite RHD are at substantial risk of progression. Although long-term outcome remains unclear, the initial change in latent RHD may be evident during the first 1 to 2 years following diagnosis. Natural history data are inherently limited, and a randomized clinical trial is needed to definitively determine the impact of penicillin prophylaxis on the trajectory of latent RHD.


Democracies restricting democratic rights: some classical sources and implications for ethics of biometrics.
Ancient Greek and 17th century English philosophy are not usually discussed along with the ethics of biometrics and data sharing. Academic ethics today, however, suffers from a lack of background in classical texts. We may discuss whether biometrics and data sharing are consistent with democracy, but if we do not know what democracy is, then we cannot know what actions are consistent with it. I shall discuss how and why democracies have restricted the rights of their citizens. I will give the most attention to two paradigms that have most influenced modern democratic thinking: 17th century English democracy and ancient Athens. I do not accept the dogma that the Athenians were obviously wrong to try and then to condemn Socrates. His death-loving doctrine could not but have weakened the will of the youth to work and fight for the good of Athens. I will try to understand the Athenians' point of view and their need to defend their security. At the end, I will apply these lessons to biometrics and data sharing for security reasons.


Implications for the use of amnion and chorion in podiatric medicine and surgery.
The purpose of this article is to show the possibility of using fetal membranes as a biologic dressing for some of the pedal ulcerations that podiatrists encounter. There are as many treatment modalities for lower extremity ulcerations as there are types of ulcers. Through the following case histories, the authors demonstrate various types of ulcerations treated with amnion and chorion and the results achieved. The clinical application of fetal membranes has been under study for many years. The authors' efforts support the use of amnion and chorion by the medical community in treating some lower extremity ulcers.


Learn, simplify and implement: developmental re-engineering strategies for cartilage repai.
The limited self-healing capacity of cartilage in adult individuals, and its tendency to deteriorate once structurally damaged, makes the search for therapeutic strategies following cartilage-related traumas relevant and urgent. To date, autologous cell-based therapies represent the most advanced treatments, but their clinical success is still hampered by the long-term tendency to form fibrous as opposed to hyaline cartilage tissue. Would the efficiency and robustness of therapies be enhanced if cartilage regeneration approaches were based on the attempt to recapitulate processes occurring during cartilage development ("developmental engineering")? And from this perspective, shouldn't cartilage repair strategies be inspired by development, but adapted to be effective in a context (an injured joint in an adult individual) that is different from the embryo ("developmental re-engineering")? Here, starting from mesenchymal stem/stromal cells (MSCs) as an adult cell source possibly resembling features of the embryonic mesenchyme, we propose a developmental re-engineering roadmap based on the following three steps: (i) learn from embryonic cartilage development which are the key pathways involved in MSC differentiation towards stable cartilage, (ii) simplify the complex developmental events by approximation to essential molecular pathways, possibly by using in vitro high-throughput models and, finally, (iii) implement the outcomes at the site of the injury by establishing an appropriate interface between the delivered signals and the recipient environment (e.g., by controlling inflammation and angiogenesis). The proposed re-design of developmental machinery by establishing artificial developmental events may offer a chance for regeneration to those tissues, like cartilage, with limited capacity to recover from injuries.


Vascular-targeting activity of ZD6126, a novel tubulin-binding agent.
The tubulin-binding agent ZD6126 is a novel vascular-targeting agent in clinical development for the treatment of solid tumors. In vivo, ZD6126 is rapidly converted into N-acetylcolchinol (ZD6126 phenol). In this study, we have explored the antivascular property of N-acetylcolchinol in vitro and ZD6126 in vivo. In cell culture, N-acetylcolchinol induced rapid changes in the morphology of human umbilical vein and lung microvessel endothelial cells. Within 40 min, the compound induced endothelial cell contraction, destabilization of the tubulin cytoskeleton, induction of actin stress fibers, and membrane blebbing. These effects occurred at noncytotoxic concentrations and were rapidly reversed on removal of the drug. Nonconfluent endothelial cells were more sensitive than confluent, quiescent cells. Among different cell types, endothelial cells were the most sensitive to the induction of morphological changes, whereas smooth muscle cells were not affected. In vitro, N-acetylcolchinol rapidly disrupted a network of newly formed cords. In vivo, ZD6126 caused shut down of newly formed vessels in the Matrigel plug assay, shortly after injection. This study indicates that rapid alteration of endothelial cell morphology may be responsible for the loss of tumor blood vessel integrity, vessel shut down, and extensive tumor necrosis induced by ZD6126 in experimental tumor models.