Distinct profiles of TERT promoter mutations and telomerase expression in head and neck cancer and cervical carcinoma.
Two recurrent mutations (-124 G > A and -146 G > A) in the core promoter region of the human telomerase reverse transcriptase (TERT) gene create consensus binding sites for ETS transcription factors and cause increased TERT expression in several tumour types. We analyzed TERT promoter mutations and TERT mRNA levels in head and neck cancer, cervical carcinoma and cervical intraepithelial neoplasia (CIN) as well as in C-4I, CaSki, HeLa and SiHa cervical cell lines. Nucleotide sequence analysis of TERT promoter region showed that 33.3% of oral squamous cell carcinoma (SCC) and 16.8% of cervical SCC harboured mutually exclusive G to A transitions at nucleotide position -124 or -146. TERT promoter was mutated at nucleotide -146 (G > A) in SiHa cell line. Other nucleotide changes creating in some cases putative ETS binding sites were more frequent in oral SCC (26.7%) than in cervical carcinoma (4.8%). The frequency of mutations was independent of human papillomavirus (HPV) tumour status in both cervical and oral cancer. Expression of TERT gene was significantly higher in TERT promoter mutated (-124G > A or -146G > A) cervical SCC compared to not mutated SCC irrespective of HPV16 E6 and E7 levels. Such hot spot changes were not detected in oropharyngeal SCC, cervical adenocarcinoma and CIN lesions. Our results suggest that TERT promoter mutations play a relevant role in oral SCC as well as in cervical SCC, besides the already known effect of HPV16 E6 protein on TERT expression.


Klotho regulates retinal pigment epithelial functions and protects against oxidative stress.
The retinal pigment epithelium (RPE) is a highly specialized CNS tissue that plays crucial roles in retinal homeostasis. Age-related morphological changes in the RPE have been associated with retinal degenerative disorders; our understanding of the underlying molecular mechanisms, however, remains incomplete. Here we report on a key role of Klotho (Kl), an aging-suppressor gene, in retinal health and RPE physiology. Kl(-/-) mice show RPE and photoreceptor degeneration, reduced pigment synthesis in the RPE, and impaired phagocytosis of the outer segment of the photoreceptors. Klotho protein (KL) is expressed in primary cultured human RPE, and regulates pigment synthesis by increasing the expression of MITF (microphthalmia transcription factor) and TYR (tyrosinase), two pivotal genes in melanogenesis. Importantly, KL increases phagocytosis in cultured RPE by inducing gene expression of MERTK/AXL/TYRO3. These effects of KL are mediated through cAMP-PKA-dependent phosphorylation of transcription factor CREB. In cultured human RPE, KL increases the l-3,4-dihydroxyphenylalanine synthesis and inhibits vascular endothelial growth factor (VEGF) secretion from basal membrane by inhibiting IGF-1 signaling and VEGF receptor 2 phosphorylation. KL also regulates the expression of stress-related genes in RPE, lowers the production of reactive oxygen species, and thereby, protects RPE from oxidative stress. Together, our results demonstrate a critical function for KL in mouse retinal health in vivo, and a protective role toward human RPE cells in vitro. We conclude that KL is an important regulator of RPE homeostasis, and propose that an age-dependent decline of KL expression may contribute to RPE degeneration and retinal pathology.


Sequential alteration of microglia and astrocytes in the rat thalamus following spinal nerve ligation.
Spinal reactive astrocytes and microglia are known to participate to the initiation and maintenance of neuropathic pain. However, whether reactive astrocytes and microglia in thalamic nuclei that process sensory-discriminative aspects of pain play a role in pain behavior remains poorly investigated. Therefore, the present study evaluated whether the presence of reactive glia (hypertrophy, increased number and upregulation of glial markers) in the ventral posterolateral thalamic nucleus (VPL) correlates with pain symptoms, 14 and 28 days after unilateral L5/L6 spinal nerve ligation (SNL) in rats. Mechanical allodynia and hyperalgesia (von Frey filament stimulation) as well as ambulatory pain (dynamic weight bearing apparatus) were assessed. Levels of nine glial transcripts were determined by quantitative real-time PCR on laser microdissected thalamic nuclei, and levels of proteins were assessed by Western blot. We also studied by immunohistofluorescence the expression of glial markers that label processes (GFAP for astrocytes and iba-1 for microglia) and cell body (S100β for astrocytes and iba-1 for microglia) and quantified the immunostained surface and the number of astrocytes and microglia (conventional counts and optical dissector method of stereological counting). Differential, time-dependent responses were observed concerning microglia and astrocytes. Specifically, at day 14, iba-1 immunostained area and number of iba-1 immunopositive cells were decreased in the VPL of SNL as compared to naïve rats. By contrast, at day 28, GFAP-immunostained area was increased in the VPL of SNL as compared to naïve rats while number of GFAP/S100β immunopositive cells remained unchanged. Using quantitative real-time PCR of laser microdissected VPL, we found a sequential increase in mRNA expression of cathepsin S (day 14), fractalkine (day 28), and fractalkine receptor (day 14), three well-known markers of microglial reactivity. Using Western blot, we confirmed an increase in protein expression of fractalkine receptor at day 14. Our results demonstrate a sequential alteration of microglia and astrocytes in the thalamus of animals with lesioned peripheral nerves. Furthermore, our data report unprecedented concomitant molecular signs of microglial activation and morphological signs of microglial decline in the thalamus of these animals.


Mobilization of Arsenic and Other Naturally Occurring Contaminants during Managed Aquifer Recharge: A Critical Review.
Population growth and climate variability highlight the need to enhance freshwater security and diversify water supplies. Subsurface storage of water in depleted aquifers is increasingly used globally to alleviate disparities in water supply and demand often caused by climate extremes including floods and droughts. Managed aquifer recharge (MAR) stores excess water supplies during wet periods via infiltration into shallow underlying aquifers or direct injection into deep aquifers for recovery during dry seasons. Additionally, MAR can be designed to improve recharge water quality, particularly in the case of soil aquifer treatment and riverbank filtration. While there are many potential benefits to MAR, introduction of recharge water can alter the native geochemical and hydrological conditions in the receiving aquifer, potentially mobilizing toxic, naturally occurring (geogenic) contaminants from sediments into groundwater where they pose a much larger threat to human and ecosystem health. On the basis of the present literature, arsenic poses the most widespread challenge at MAR sites due to its ubiquity in subsurface sediments and toxicity at trace concentrations. Other geogenic contaminants of concern include fluoride, molybdenum, manganese, and iron. Water quality degradation threatens the viability of some MAR projects with several sites abandoning operations due to arsenic or other contaminant mobilization. Here, we provide a critical review of studies that have uncovered the geochemical and hydrological mechanisms controlling mobilization of arsenic and other geogenic contaminants at MAR sites worldwide, including both infiltration and injection sites. These mechanisms were evaluated based on site-specific characteristics, including hydrological setting, native aquifer geochemistry, and operational site parameters (e.g., source of recharge water and recharge/recovery cycling). Observed mechanisms of geogenic contaminant mobilization during MAR via injection include shifting redox conditions and, to a lesser extent, pH-promoted desorption, mineral solubility, and competitive ligand exchange. The relative importance of these mechanisms depends on various site-specific, operational parameters, including pretreatment of injection water and duration of injection, storage, and recovery phases. This critical review synthesizes findings across case studies in various geochemical, hydrological, and operational settings to better understand controls on arsenic and other geogenic contaminant mobilization and inform the planning and design of future MAR projects to protect groundwater quality. This critical review concludes with an evaluation of proposed management strategies for geogenic contaminants and identification of knowledge gaps regarding fate and transport of geogenic contaminants during MAR.


Diabetes in urban African-Americans. XVII. Availability of rapid HbA1c measurements enhances clinical decision-making.
To assess the impact of rapid-turnaround HbA1c results on providers' clinical decision-making and on follow-up HbA1c levels. The research design was a randomized clinical trial in which rapid HbA1c results were made available to providers on even days of the month (rapid, n = 575), but delayed by 24 h on odd days (conventional, n = 563). Adjustment of therapy for patients with type 2 diabetes was considered appropriate if therapy was intensified for HbA1c values >7% or not intensified for HbA1c values < or =7%. A post-hoc analysis was also performed using patients (n = 574) who returned for follow-up 2-7 months later to ascertain the effect of rapid HbA1c availability on subsequent glycemic control. Rapid HbA1c availability resulted in more appropriate management compared with conventional HbA1c availability (79 vs. 71%, P = 0.003). This difference was due mainly to less frequent intensification when HbA1c levels were < or =7% (10 vs. 22%, P < 0.0001) and slightly to more frequent intensification for patients with HbA1c values >7% (67 vs. 63%, P = 0.33). For both groups, intensification was greatest for patients on insulin (51%) compared with patients on oral agents (35%) and diet alone (14%) (P < 0.0001). Regression analysis confirmed that providers receiving conventional HbA1c results were more likely to intensify therapy in patients who already had HbA1c levels < or =7%. Over 2-7 months of follow-up, HbA1c rose more in patients with conventional HbA1c results compared with rapid results (0.8 vs. 0.4%, P = 0.02). In patients with initial HbA1c >7%, rapid HbA1c results had a favorable impact on follow-up HbA1c independent of the decision to intensify therapy (P = 0.03). Availability of rapid HbA1c determinations appears to facilitate diabetes management. The more favorable follow-up HbA1c profile in the rapid HbA1c group occurs independently of the decision to intensify therapy, suggesting the involvement of other factors such as enhanced provider and/or patient motivation.


A Meta-Analysis on Degraded Alpine Grassland Mediated by Climate Factors: Enlightenment for Ecological Restoration.
Alpine grassland is the main ecosystem on the Qinghai-Tibet Plateau (QTP). Degradation and restoration of alpine grassland are related to ecosystem function and production, livelihood, and wellbeing of local people. Although a large number of studies research degraded alpine grassland, there are debates about degradation patterns of alpine grassland in different areas and widely applicable ecological restoration schemes due to the huge area of the QTP. In this study, we used the meta-analysis method to synthesize 80 individual published studies which were conducted to examine aboveground and underground characteristics in non-degradation (ND), light degradation (LD), moderate degradation (MD), heavy degradation (HD), and extreme degradation (ED) of alpine grassland on the QTP. Results showed that aboveground biomass (AGB), belowground biomass (BGB), Shannon-Wiener index (H'), soil moisture (SM), soil organic carbon (SOC), soil total nitrogen (TN), and available nitrogen (AN) gradually decreased along the degradation gradient, whereas soil bulk density (BD) and soil pH gradually increased. In spite of a tendency to soil desertification, losses of other soil nutrients and reduction of enzymes, there was no linear relationship between the variations with degradation gradient. Moreover, the decreasing extent of TN was smaller in areas with higher precipitation and temperature, and the decreasing extent of AGB, SOC, and TN was larger in areas with a higher extent of corresponding variables in the stage of ND during alpine grassland degradation. These findings suggest that in areas with higher precipitation and temperature, reseeding and sward cleavage can be used for restoration on degraded alpine grassland. Fencing and fertilization can be used for alpine grassland restoration in areas with lower precipitation and temperature. Microbial enzymes should not be used to restore degraded alpine grassland on a large scale on the QTP without detailed investigation and analysis. Future studies should pay more attention to the effects of climate factors on degradation processes and specific ecological restoration strategies in different regions of the QTP.


The problem with cationic liposome/micelle-based non-viral vector systems for gene therapy.
Gene therapy research is in crisis owing to the lack of acceptable vector systems to deliver nucleic acids to patients for therapy. Viral vectors are efficient but currently appear to be too dangerous for routine clinical use. Synthetic non-viral vectors are inherently much safer but are currently not efficient enough to be clinically viable. The solution for gene therapy lies with improved synthetic non-viral vectors based upon well-found platform technologies and a thorough understanding of the barriers to efficient gene delivery and expression (transfection) relevant to clinical applications of interest. In this review, the current status and prospects for cationic liposome/micelle-based synthetic non-viral vector systems are discussed including a description of the barriers to efficient transfection, a summary of the main structure/activity studies and mention of ternary cationic liposome/micelle-nucleic acid (LD) systems. The review culminates with a description of two promising cationic liposome/micelle-based non-viral vector platform systems known as liposome:mu:DNA (LMD) and stabilised plasmid-lipid particles (SPLP) that should create a real opportunity for the development of clinically viable synthetic vector systems within the next few years.


Recognizing emotions expressed by body pose: a biologically inspired neural model.
Research into the visual perception of human emotion has traditionally focused on the facial expression of emotions. Recently researchers have turned to the more challenging field of emotional body language, i.e. emotion expression through body pose and motion. In this work, we approach recognition of basic emotional categories from a computational perspective. In keeping with recent computational models of the visual cortex, we construct a biologically plausible hierarchy of neural detectors, which can discriminate seven basic emotional states from static views of associated body poses. The model is evaluated against human test subjects on a recent set of stimuli manufactured for research on emotional body language.


Experimental investigation on removal of suspended particles from water using flax fibre geotextiles.
Natural geotextiles are increasingly used in geotechnical applications such as bank protection and short-term soil reinforcement. This study aimed to highlight the behaviour of natural flax fibre geotextiles towards the retention of suspended particles (SP) present in urban runoff and often polluted. Indeed, it is well known that a large fraction of the heavy metals are often associated with the SP. Long-term filtration tests were performed on four laboratory filter designs. The objective was to study the influence of a nonwoven flax fibre geotextiles on SP transfer through a sandy porous media under saturated conditions. The experiments consisted of injecting SP in a column filled with sand with and without flax fibre geotextiles at a constant flow rate. The results showed that nonwoven flax fibre geotextiles can increase flow homogeneity. Retention efficiency of SP at column scale as well as spatial deposition profile was modified by the presence of geotextile. The use of flax fibre geotextiles increases considerably the durability of the filtration system. Particle size analysis of the particles retained in the sand medium and in geotextiles shows that the coarser particles are mainly deposited at the entrance of the column, and in the geotextiles.


Glutamate oxidase sheets-Prussian blue grafted amperometric biosensor for the real time monitoring of glutamate release from primary cortical neurons.
Glutamate (GLU) is a primary excitatory neurotransmitter, and its dysregulation is associated with several neurodegenerative disorders. A major challenge in GLU estimation is the existence of other biomolecules in the brain that could directly get oxidized at the electrode. Hence, highly selective electroenzymatic biosensors that enable rapid estimation of GLU are needed. Initially, a copolymer, poly(2-dimethylaminoethyl methacrylate- styrene) was synthesized through reversible addition-fragmentation chain transfer polymerization to noncovalently functionalize reduced graphene oxide (rGO), named DS-rGO. Glutamate oxidase macromolecule immobilized DS-rGO formed enzyme nanosheets, which was drop-coated over Prussian blue electrodeposited disposable electrodes to fabricate the GLU biosensor. The interconnectivity between the enzyme nanosheets and the Prussian blue endows the biosensor with enhanced conductivity and electrochemical activity. The biosensor exhibited a linearity: 3.25-250 μM; sensitivity: 3.96 μA mM-1 cm-2, and a limit of detection: 0.96 μM for GLU in the Neurobasal Medium. The biosensor was applied to an in vitro primary rat cortical model to discriminate GLU levels in Neurobasal Medium, before and after KCl mediated depolarization, which provides new insights for elucidating neuronal functioning in the brain.


Sonochemical synthesis of lignin nanoparticles and their applications in poly (vinyl) alcohol composites.
Lignin is a common and abundant byproduct of the pulp and paper industry and is generally burned to produce steam. Opportunities exist to acquire greater value from lignin by leveraging the properties of this highly conjugated biomacromolecule for applications in UV absorption and polymer reinforcement. These applications can be commercialized by producing value-added lignin nanoparticles (LNPs) using a scalable sonochemical process. In the present research, monodisperse LNPs have been synthesized by subjecting aqueous dispersions of alkali lignin to acoustic irradiation. The resulting particle size distribution and colloidal stability, as determined by dynamic light scattering, transmission electron microscopy and zeta potential analysis, of LNPs can be adjusted by varying the solution pH and ultrasonication energy. As-synthesized LNPs with a mean diameter of 204 nm were incorporated into poly (vinyl) alcohol (PVA) to prepare thin and flexible nanocomposite films using a simple solvent casting method. The addition of 2.5 wt% LNP increased the material's Sun Protection Factor up to 26 compared to 0 for neat PVA, while maintaining light transmission above 75 % in the visible spectra. In addition, the tensile strength and elastic modulus of the PVA nanocomposites improved by 47 % and 36 %, respectively. The presence of LNP also enhanced the thermal stability of the materials. Significantly, the proposed sonochemical process may be generally applicable to the synthesis of a range of naturally-derived LNPs for a variety of value-added applications.


Effects of global competition on health care administration in Finland and the United States. What you can do to save your practice.
Global competition, a faltering Finnish economy, and deep recession are forcing difficult decisions on the health care administrators of this Nordic country known for its extensive social welfare safety net and socialized health care system. In the first part of this article the Finnish health care system and its problems, as well as possible solutions, are presented. Similarities to our own system are noted and discussed. In an interview, the director general for the Minister of Social Affairs and Health of Finland, Kimmo Leppo, expressed concern about future funding sources in his country for the same levels of social welfare and health care programs that presently exist. The Finns are considering managed competition among providers, increasing copayments, excess hospital beds, anticipated hospital closures in their country, block grants, and how to deal with rising physician unemployment. He was interested in the proposed US government health care plans, and especially the state of Tennessee's TennCare plan, a state plan that closely resembled the failed Clinton health care reform package. The second part of this article explores how global competition has brought the pressure of modern competitive economics into the US medical arena.


Size at onset of sexual maturity in Macrobrachium amazonicum (Heller, 1862) phenotypes: an integrative approach.
The size at the onset of sexual maturity (SOM) was determined in four hololimnetic populations of Macrobrachium amazonicum using morphometric, physiological and functional criteria. Male prawns from two populations analyzed had hypertrophied chelipeds and large body proportions, showing the presence of four morphotypes, i.e. large-size phenotype (LS). However, the other two populations showed smaller males without morphotypes, i.e. small-size phenotype (SS). The development of sexual weapons in males modulated the mating system and SOM in this species. It was verified that there is a synchrony between physiological and functional maturities in SS males. On the other hand, functional maturity in LS males occurred after physiological and morphological maturities. In females, we verified synchronization between morphological and functional maturities. The results obtained indicated that, in both sexes, physiological maturity anticipates the others criteria. There were no differences between the sequential scheme of sexual maturity for female phenotypes, being: physiological → functional morphological. However, the evaluation of sexual maturity in males must take into account the phenotype of population, since that was modulated by functional maturity. The sequential scheme for LS males is: physiological → morphological → functional, while for SS males it is: physiological = functional → morphological.


A questionnaire for assessing environmental tobacco smoke exposure.
Exposure to environmental tobacco smoke (ETS) has been associated with adverse health consequences. In this study, the association between self-reported ETS exposure and serum cotinine levels was examined. As part of the population-based Epidemiology of Hearing Loss Study in Beaver Dam, WI, self-reported data on ETS were obtained at the 5-year follow-up examination (1998-2000; n = 2800). Serum cotinine levels were obtained on 643 of these participants (53-75 years old). Serum cotinine levels increased with reported number of hours of recent ETS exposure. Most (95.3%) participants who reported being smokers had serum cotinine levels >15 ng/mL, compared with only 2.7% of nonsmokers. Usual ETS exposure among nonsmokers was assessed with a brief questionnaire quantifying exposure at work, at home, and in social settings, and then classified into three levels of exposure. Those reporting little or no exposure had a geometric mean cotinine level of 0.06 ng/mL (95% confidence interval (CI): 0.05, 0.07), compared with 0.14 (95% CI: 0.09, 0.22) and 0.63 (95% CI: 0.36, 1.10) for those reporting moderate and high ETS exposure, respectively (F-test for trend: P < 0.001). These results suggest that the questionnaire can be used to distinguish relative levels of exposure to ETS.


Chemokines in the balance: maintenance of homeostasis and protection at CNS barriers.
In the adult central nervous system (CNS), chemokines and their receptors are involved in developmental, physiological and pathological processes. Although most lines of investigation focus on their ability to induce the migration of cells, recent studies indicate that chemokines also promote cellular interactions and activate signaling pathways that maintain CNS homeostatic functions. Many homeostatic chemokines are expressed on the vasculature of the blood brain barrier (BBB) including CXCL12, CCL19, CCL20, and CCL21. While endothelial cell expression of these chemokines is known to regulate the entry of leukocytes into the CNS during immunosurveillance, new data indicate that CXCL12 is also involved in diverse cellular activities including adult neurogenesis and neuronal survival, having an opposing role to the homeostatic chemokine, CXCL14, which appears to regulate synaptic inputs to neural precursors. Neuronal expression of CX3CL1, yet another homeostatic chemokine that promotes neuronal survival and communication with microglia, is partly regulated by CXCL12. Regulation of CXCL12 is unique in that it may regulate its own expression levels via binding to its scavenger receptor CXCR7/ACKR3. In this review, we explore the diverse roles of these and other homeostatic chemokines expressed within the CNS, including the possible implications of their dysfunction as a cause of neurologic disease.


Measuring positive and negative aspects of youth behavior: Development and validation of the Adolescent Functioning Scale.
This paper outlines the development and validation of the Adolescent Functioning Scale (AFS) in an Australian sample of parents of young people aged 11-18 years (N = 278). The AFS, a parent self-report measure, was designed to assess problem behavior and positive development in adolescents. Principal components analysis produced a 33-item measure comprising four subscales: Positive Development, Oppositional Defiant Behavior, Antisocial Behavior and Emotional Difficulties. Convergent validity was established via correlations between the AFS and established measures of adolescent functioning and parenting, and discriminant validity was shown through no association between the AFS and a measure of technology use. Internal consistency for the subscales was high (H = .82-.92 for different age groups), as was test-retest reliability (r = .77-.86). The study indicated that the AFS is a potentially valuable tool for assessing levels of problem behaviors and positive development in adolescents.


[Prevalence and pharmacotherapy of diabetes mellitus in primary care].
So far only incomplete epidemiological data on the management of diabetes mellitus have been available for the Federal Republic of Germany. It was the aim of this study to obtain such information from a representative cross-section of patients. 43549 consecutive, unselected patients (52.6% females; mean age 64.9 +/- 11.7 years) of general practitioners and internists in private practice were included in this study which was based on data provided by the general practitioners or internists during September 2001. They recorded prevalence of diabetes, hypertension and 22 other diseases, and the patients' current treatment. Also recorded were associations of other patient-related variables (age, gender, associated or resulting diseases, hypertension, micro- and macrovascular complications, obesity), with medical practitioner-related variables (medical training, use of guidelines, regional factors) and with the frequency of antidiabetic treatment (total and differentiated by class of drug). The prevalence of diabetes mellitus in this cohort was 15.6% (18.5% for males and 13.7% for females). 67.6% of the diabetics received antidiabetic medication. There was little difference, between the different age groups and between males and females, with regard to which of the various drugs were prescribed. Patients with macrovascular complications were more intensively treated with antidiabetics than those with exclusively microvascular ones. There was hardly any correlation between patterns of prescription and various practitioner characteristics (medical training, use of guidelines, regional factors). There was little differentiation in the prescription of antidiabetic medication by general practitioners/internists among an unselected patient cohort.


Pediatric Internet resources. Creation and growth of the PEDINFO index.
To illustrate the growth of pediatric-related material on the Internet as demonstrated by the growth and use of an index of pediatric Internet resources. Descriptive analysis. The log files of the computer systems on which the PEDINFO index was implemented were examined. File size of the index was measured, and a record of Internet hosts that connected to the server each day was produced. A proportion of Internet sites (n = 300) were examined for author reliability and practice enhancement value. In a period of 14 months, the file size grew from 7 kilobytes (KB) to more than 80 KB. By November 1995, usage exceeded 250 individuals per day and has since then leveled off to about 180 users per day. Analysis of the domains of the users showed a shift from equal proportion of "com" (commercially obtained Internet addresses) and "edu" (educational institution) addresses to an increase in com addresses with a stable proportion of edu addresses. One hundred twenty-seven sites contained patient or parent information, and 62 sites contained reference material. There was a wide range of author reliability ratings. From the growth of PEDINFO, we conclude that an increase in pediatric-related information available on the Internet is steady, much of which is patient or parent educational material. We anticipate further growth and use of the Internet in the exchange of information and cell for further education regarding its use so that pediatricians can more easily direct their patients to the most medically relevant sources.


The transcription factor HNF1α regulates expression of chloride-proton exchanger ClC-5 in the renal proximal tubule.
The Cl(-)/H(+) exchanger ClC-5 is essential for the endocytic activity of the proximal tubule cells and the tubular clearance of proteins filtered in the glomeruli. The mechanisms that regulate the expression of ClC-5 in general and its specific expression in the proximal tubule are unknown. In this study, we investigated the hypothesis that the hepatocyte nuclear transcription factor HNF1α, which is predominantly expressed in proximal tubule segments, may directly regulate the expression of ClC-5. In situ hybridization demonstrated that the expression of Clcn5 overlaps with that of Hnf1α in the developing kidney as well as in absorptive epithelia, including the digestive tract and yolk sac. Multiple binding sites for HNF1 were mapped in the 5'-regulatory sequences of the mouse and human Clcn5/CLCN5 genes. The transactivation of the Clcn5/CLCN5 promoter by HNF1α was verified in vitro, and the binding of HNF1α to the Clcn5 promoter in vivo was confirmed by chromatin immunoprecipitation in mouse kidney. The expression of Clcn5 was reduced in the proximal tubule segments of HNF1α-null kidneys, and it was rescued upon transfection of HNF1α-null cells with wild-type but not with mutant HNF1α. These data demonstrate that HNF1α directly regulates the expression of ClC-5 in the renal proximal tubule and yield insights into the mechanisms governing epithelial differentiation and specialized transport activities in the kidney.


Measurement of pulse reappearance time in diagnosis of peripheral vascular disease in diabetes.
Forty-eight patients (20 diabetic, 28 nondiabetic) with angiographically confirmed peripheral vascular disease (PVD) were examined to discover whether the measurement of pulse reappearance time (PRT) during reactive hyperemia is a more useful method than the measurement of peripheral systolic blood pressure (ankle pressure index; API) for making a specific diagnosis of PVD. Specific diagnosis refers to the degree and localization of occlusive atherosclerosis determined by Doppler ultrasound techniques for both measurements. We found that PRT and API both provided accurate qualitative proof of a peripheral blood flow deficit in diabetic and nondiabetic subjects. However, in relation to the angiographically defined degree and localization of sclerotic lesions, there were significant differences. The sclerotic degree of occlusive PVD in diabetic subjects was correlated with the results of the PRT (P less than .001), whereas the API was not (P greater than .05). The occlusion localization could only be distinguished by PRT measurements in both diabetic and nondiabetic subjects. Compared with control subjects (4.1 s) the half-maximum PRT of blood flow velocity was delayed in stenotic PVD to 5.7 s, in occlusive PVD of the upper leg to 14.3 s, in occlusive PVD of the lower leg to 29.6 s, and in multilevel disease to 45.0 s (P less than .0005 vs. control). The results show that Doppler sonographic measurement of the peripheral systolic blood pressure is only useful for an overall diagnosis of PVD in diabetic subjects, whereas PRT measurement, by quantifying the degree and localization of sclerotic lesions, can be used additionally either to confirm or to specify this diagnosis.


Costs of medication use among patients with juvenile idiopathic arthritis in the Dutch healthcare system.
Background: This study aims to quantify medication costs in juvenile idiopathic arthritis (JIA), based on subtype.Research design and methods: This study is a single-center, retrospective analysis of prospective data from electronic medical records of JIA patients, aged 0-18 years between 1 April 2011 and 31 March 2019. Patient characteristics (age, gender, subtype) and medication use were extracted. Medication use and costs were reported as: 1) mean total annual costs; 2) between-patient heterogeneity in these costs; 3) duration of medication use; and, 4) costs over the treatment course.Results: The analysis included 691 patients. Mean total medication costs were €2,103/patient/year, including €1,930/patient/year (91.8%) spent on biologicals. Costs varied considerably between subtypes, with polyarticular rheumatoid-factor positive and systemic JIA patients having the highest mean costs (€5,020/patient/year and €4,790/patient/year, respectively). Mean annual medication costs over the patient's treatment course ranged from <€1,000/year (71.1% of patients) to >€11,000/year (2.5% of patients). Etanercept and adalimumab were the most commonly used biologicals. Cost fluctuations over the treatment course were primarily attributable to biological use.Conclusions: Polyarticular rheumatoid-factor positive and systemic JIA patients had the highest mean total annual medication costs, primarily attributable to biologicals. Costs varied considerably between subtypes, individuals, and over the treatment course.


Use of a saline chaser to reduce the dose of contrast material for dynamic CT in cattle.
To investigate the effectiveness of a saline chaser in reducing the required dose of contrast material while carrying out dynamic CT scans in cattle, six clinically normal Holstein calves were subjected to CT scans of the head in a study using a crossover design. The calves were studied as three groups: group A received only contrast material (600 mgI/kg); group B received a 30 per cent lower dose of the same contrast material followed by 50 ml of saline solution; and group C received a 40 per cent lower dose of the same contrast material followed by 50 ml of saline solution. In all the animals, a power injector was used for administering the contrast material, with an injection rate of 4 ml/s and a pressure of 4.7 to 5.7 kg/cm(2). Attenuation values were obtained from the right and left maxillary arteries and dorsal sagittal sinus. There were no significant differences in the maximum enhancement value (MEV) of the maxillary arteries and dorsal sagittal sinus between groups A and B. The MEVs of the maxillary arteries and dorsal sagittal sinus in groups A and B were significantly greater than the MEV in group C (P<0.05). The use of a saline chaser allowed a 30 per cent reduction in the dose of contrast material without decreasing vascular enhancement.


Incongruous consultation behaviour: results from a UK-wide population survey.
Symptom characteristics are strong drivers of care seeking. Despite this, incongruous consultation behaviour occurs and has implications for both individuals and health-care services. The aim of this study was to determine how frequently incongruous consultation behaviour occurs, to examine whether it is more common for certain types of symptoms and to identify the factors associated with being an incongruous consulter. An age and sex stratified random sample of 8,000 adults was drawn from twenty UK general practices. A postal questionnaire was used to collect detailed information on the presence and characteristics of 25 physical and psychological symptoms, actions taken to manage the symptoms, general health, attitudes to symptom management and demographic/socio-economic details. Two types of incongruous consultation behaviour were examined: i) consultation with a GP for symptoms self-rated as low impact and ii) no consultation with a GP for symptoms self-rated as high impact. A fifth of all symptoms experienced resulted in consultation behaviour which was incongruous based on respondents' own rating of the symptoms' impact. Low impact consultations were not common, although symptoms indicative of a potentially serious condition resulted in a higher proportion of low impact consultations. High impact non-consultations were more common, although there was no clear pattern in the type of associated symptoms. Just under half of those experiencing symptoms in the previous two weeks were categorised as an incongruous consulter (low impact consulter: 8.3%, high impact non-consulter: 37.1%). Employment status, having a chronic condition, poor health, and feeling that reassurance or advice from a health professional is important were associated with being a low impact consulter. Younger age, employment status, being an ex-smoker, poor health and feeling that not wasting the GPs time is important were associated with being a high impact non-consulter. This is one of the first studies to examine incongruous consultation behaviour for a range of symptoms. High impact non-consultations were common and may have important health implications, particularly for symptoms indicative of serious disease. More research is now needed to examine incongruous consultation behaviour and its impact on both the public's health and health service use.


Individual and health system variation in rehospitalizations the year after pneumonia.
Little is known about variation in patterns of recovery among patients discharged alive from hospitalizations for pneumonia.The aim of the is observational cohort study was to characterize the variation in patterns of hospital readmission and survival in the year after discharge for pneumonia in 3 different health systems.The 3 cohorts consisted of (1) the Health and Retirement Study participants enrolled in Fee-for-service Medicare (FFS), (2) Veterans Administration (VA) Healthcare system, and (3) Kaiser Permanente of Northern California (KPNC). The 365-day survival and re-hospitalizations were determined for each cohort. Multinomial logistic regression was used to identify potential contributors to the different patterns.We identified 2731, 23,536, and 39,147 hospitalizations for pneumonia in FFS Medicare, VA, and KPNC, respectively, of whom 88.1%, 92.8%, and 89.7% survived to hospital discharge. The median patient survived to 1 year and was rehospitalized twice in FFS (9.0%), once in VA (14.1%) and KPNC (9.1%). Of the patients who survived the hospitalization, 33.3% (FFS), 30.2% (VA), and 26.8% (KPNC) died during the subsequent year. Of those who survived, 29.8% (FFS), 35.9% (VA), and 46.1% (KPNC) were never rehospitalized. 11.9% (FFS), 11.9% (VA), and 11.7% (KPNC) had greater than 3 hospitalizations. Age, race, gender, comorbidity, ICU use, and hospital length stay collectively explained little (5-7%) of the variation in the recovery pattern.There is significant variation in the year after the hospitalization for pneumonia across individuals, but less so across health systems. There may be important opportunities to better classify these heterogeneous individual-level pathways.


The Personalized Advantage Index: translating research on prediction into individualized treatment recommendations. A demonstration.
Advances in personalized medicine require the identification of variables that predict differential response to treatments as well as the development and refinement of methods to transform predictive information into actionable recommendations. To illustrate and test a new method for integrating predictive information to aid in treatment selection, using data from a randomized treatment comparison. Data from a trial of antidepressant medications (N = 104) versus cognitive behavioral therapy (N = 50) for Major Depressive Disorder were used to produce predictions of post-treatment scores on the Hamilton Rating Scale for Depression (HRSD) in each of the two treatments for each of the 154 patients. The patient's own data were not used in the models that yielded these predictions. Five pre-randomization variables that predicted differential response (marital status, employment status, life events, comorbid personality disorder, and prior medication trials) were included in regression models, permitting the calculation of each patient's Personalized Advantage Index (PAI), in HRSD units. For 60% of the sample a clinically meaningful advantage (PAI≥3) was predicted for one of the treatments, relative to the other. When these patients were divided into those randomly assigned to their "Optimal" treatment versus those assigned to their "Non-optimal" treatment, outcomes in the former group were superior (d = 0.58, 95% CI .17-1.01). This approach to treatment selection, implemented in the context of two equally effective treatments, yielded effects that, if obtained prospectively, would rival those routinely observed in comparisons of active versus control treatments.


Should this patient be screened for cancer?
Advances in imaging technology have provided numerous opportunities for cancer screening but have also raised numerous questions. Who should be screened and how exactly should screening be performed? If spiral computed tomography (spiral CT) were being considered for lung cancer screening, for example, important questions would need to be answered: Should nonsmokers be screened? How often should screening take place? What should the diagnostic work-up be after abnormal findings were seen on spiral CT? Randomized, controlled trials (RCTs) are the most valid method for determining which medical interventions are most effective. These trials are particularly useful in the evaluation of screening because they eliminate the early detection biases that may result in groosly misleading survival statistics. Randomized, controlled trials of screening are subject to other biases, and their results may be difficult to generalize. In addition, because they require an enormous number of participants and many years of follow-up, RCTs can be applied only to a small proportion of the questions about cancer screening. Quantitative decision analysis can be applied to the remaining questions and help inform decision making about cancer screening.


Parkinson's disease: focus on management alternatives.
This activity is designed for physicians, pharmacists, nurses, health planners, directors of managed care organizations, and payers of health services. To understand the impact that Parkinson's disease has on patients and to identify areas of drug therapy that can be optimized to improve a patient's quality of life. 1. Discuss the epidemiology and pathophysiology of idiopathic Parkinson's disease (IPD). 2. Identify causes of secondary parkinsonism and medications associated with drug-induced parkinsonism. 3. Discuss the cardinal and secondary manifestations of IPD. 4. Outline the 6 stages of IPD. 5. Identify therapeutic alternatives for various levodopa treatment failures. 6. Identify the 3 newer antiparkinson agents and state their mechanisms of action, common adverse effects, drug interactions, and appropriate use. 7. Identify the appropriate use, common adverse effects, and drug interactions of dopamine agonists, anticholinergic agents, monoamine oxidase inhibitors, and catechol-O-methyltransferase inhibitors in Parkinson's disease therapy.


Synthesis and characterization of nanocrystalline calcium sulfate for use in osseous regeneration.
Nanoparticles of calcium sulfate (nCS) have potential advantages as a ceramic matrix, scaffold and/or vehicle for delivering growth factors for osseous regeneration in a variety of clinical situations. The objectives of this study were to synthesize and characterize nanoparticles of hemihydrate calcium sulfate (nCS) and to develop a nCS-based system for bone regeneration. A cryo-vacuum method was used to process dihydrate CS into dihydrate nCS, which was then subjected to oven drying to produce hemihydrate. The nCS was sterilized by glow discharge treatment for use as a synthetic graft material for the treatment of bone defects. Electron microscopy showed that the nCS powder consisted of aggregates of closely arranged acicular crystals, approximately 30-80 nm in width, 400-600 nm in length and approximately 80-100 nm in diameter, providing a surface area about ten times that of conventional CS. Thorough physico-chemical characterization confirmed the composition and phase of the material. Cell viability/metabolic activity assays and alkaline phosphate assays confirmed the safety and biocompatibility of nCS. Release kinetics for adsorbed platelet-derived growth factor and bone morphogenetic protein-2 (BMP-2) suggests that nCS may serve as an appropriate vehicle for slow release delivery of these agents. The studies presented here give evidence of the advantages of nCS as a scaffold to support osteoblastic cell activity.


Pathogenesis of RSV lower respiratory tract infection: implications for vaccine development.
Respiratory syncytial virus (RSV) infection is the most prevalent cause of severe respiratory disease in infants. It also causes considerable morbidity in older children and adults with underlying risk factors. RSV vaccine development has been complicated by the need to administer the vaccine at a very young age and by enhanced disease observed after vaccination with formalin inactivated RSV. For infants live attenuated vaccines, which may not be expected to predispose for vaccine induced enhanced pathology, hold the greatest promise. However, the balance between attenuation and immunogenicity appears to be delicate. For older risk groups, results with subunit vaccines are most promising.


Diversity of microbial communities and soil nutrients in sugarcane rhizosphere soil under water soluble fertilizer.
The dynamics of soil microbial communities are important for plant health and productivity. Soil microbial communities respond differently to fertilization. Organic water soluble fertilizer is an effective soil improver, which can effectively improve soil nutrient status and adjust soil pH value. However, little is known about the effects of water soluble fertilizers on soil microbial community, and the combined effects on soil nutrients and sugarcane productivity. Therefore, this study sought to assess the effects of water soluble fertilizer (1,050 kg/hm2 (WS1), 1,650 kg/hm2 (WS2)) and mineral fertilizer (1,500 kg/hm2 (CK)) on the soil microbial community, soil nutrients and crop yield of sugarcane. The results showed that compared with CK, the application of water soluble fertilizers (WS1 and WS2) alleviated soil acidity, increased the OM, DOC, and AK contents in the soil, and further improved agronomic parameters and sugarcane yield. Both WS1 and WS2 treatments significantly increased the species richness of microorganisms, especially the enrichment of beneficial symbiotic bacteria such as Acidobacteria and Planctomycetes, which are more conducive to the healthy growth of plants. Furthermore, we found that soil nutrient contents were associated with soil microbial enrichment. These results indicate that water soluble fertilizer affects the enrichment of microorganisms by improving the nutrient content of the soil, thereby affecting the growth and yield of sugarcane. These findings therefore suggest that the utilization of water soluble fertilizer is an effective agriculture approach to improve soil fertility.


A serious adverse surgical event: Management of suspected HSV-1 keratitis in a donor cornea.
To describe the management of a serious adverse event in a patient undergoing penetrating keratoplasty (PK). A 68-year-old man underwent PK for an aphakic bullous keratopathy following previous complicated cataract surgery. He had no past history of herpetic disease. Storage of the corneoscleral disc in the transport bottle precluded microscopic examination. After placement of the trephined donor cornea on the open eye of the recipient, a large dendritiform geographic ulcer was noted on the donor cornea. A replacement cornea was used after changing potentially contaminated instruments. Intravenous antiviral treatment was commenced intraoperatively to reduce the risk of infection to the central nervous system. Postoperatively, oral and topical antiviral treatment was commenced and 6 months following surgery the patient developed a geographic corneal ulcer at the graft host interface. Containers to transport corneoscleral discs should enable microscopic examination by the surgeon prior to use. High dose systemic antivirals may reduce the risk of herpetic disease involving the posterior segment of the eye and neuroretina in the aphakic eye and spread to the central nervous system.


Cholinesterases as biomarkers for parasympathetic dysfunction and inflammation-related disease.
Accumulating evidence suggests parasympathetic dysfunction and elevated inflammation as underlying processes in multiple peripheral and neurological diseases. Acetylcholine, the main parasympathetic neurotransmitter and inflammation regulator, is hydrolyzed by the two closely homologous enzymes, acetylcholinesterase and butyrylcholinesterase (AChE and BChE, respectively), which are also expressed in the serum. Here, we consider the potential value of both enzymes as possible biomarkers in diseases associated with parasympathetic malfunctioning. We cover the modulations of cholinesterase activities in inflammation-related events as well as by cholinesterase-targeted microRNAs. We further discuss epigenetic control over cholinesterase gene expression and the impact of single-nucleotide polymorphisms on the corresponding physiological and pathological processes. In particular, we focus on measurements of circulation cholinesterases as a readily quantifiable readout for changes in the sympathetic/parasympathetic balance and the implications of changes in this readout in health and disease. Taken together, this cumulative know-how calls for expanding the use of cholinesterase activity measurements for both basic research and as a clinical assessment tool.


Impaired tethering and fusion of GLUT4 vesicles in insulin-resistant human adipose cells.
Systemic glucose homeostasis is profoundly influenced by adipose cell function. Here we investigated GLUT4 dynamics in living adipose cells from human subjects with varying BMI and insulin sensitivity index (Si) values. Cells were transfected with hemagglutinin (HA)-GLUT4-green fluorescent protein (GFP)/mCherry (red fluorescence), and were imaged live using total internal reflection fluorescence and confocal microscopy. HA-GLUT4-GFP redistribution to the plasma membrane (PM) was quantified by surface-exposed HA epitope. In the basal state, GLUT4 storage vesicle (GSV) trafficking to and fusion with the PM were invariant with donor subject Si, as was total cell-surface GLUT4. In cells from insulin-sensitive subjects, insulin augmented GSV tethering and fusion approximately threefold, resulting in a corresponding increase in total PM GLUT4. However, with decreasing Si, these effects diminished progressively. All insulin-induced effects on GLUT4 redistribution and trafficking correlated strongly with Si and only weakly with BMI. Thus, while basal GLUT4 dynamics and total cell-surface GLUT4 are intact in human adipose cells, independent of donor Si, cells from insulin-resistant donors show markedly impaired GSV tethering and fusion responses to insulin, even after overnight culture. This altered insulin responsiveness is consistent with the hypothesis that adipose cellular dysfunction is a primary contributor to systemic metabolic dysfunction.


[Cytomorphological and metabolic changes in the adrenal glands of rats exposed to coal and rock dust and during exercise].
Cytomorphological and cytochemical studies were used to examine 28 experimental non-inbred male rats that had been intratracheally exposed to 50 mg of coal rock dust (CRD) and had concurrently exercises (running) for 2 hours 5 times a week during a 3-month experiment. Toxic and physical factors were found to adversely affect the functional status of the sympathoadrenal systems of adrenal cortical cells (ACC). The most significant impairments of cytomorphological and metabolic changes were observed in ACC under the combined effects of CRD and during exercise.


[Evaluation of cardio-pulmonary function for lobectomy under the condition of selective pulmonary arterial occlusion test (SPAO)].
We performed selective pulmonary arterial occlusion test (SPAO) and unilateral pulmonary arterial occlusion test (UPAO) in 30 patients with lung cancer who would be undertaken right lower lobectomy or middle and lower lobectomy, and evaluated the preoperative cardio-pulmonary function. The lung hemodynamics during SPAO were different from those during UPAO. This indicates that SPAO evaluated reserve capacity of vascular bed in the right upper lobe which would be remained at the surgery. Moreover, the ratio of total pulmonary vascular resistance during SPAO to that of before SPAO was not differed in two groups, one was a group with and the other was a group without impaired reserve capacity of vascular bed in the contralateral lung. This suggests that reserve capacity of pulmonary vascular bed including right upper lobe would be well compensated even in the patients with highly impaired capacity in the contralateral lung. Based on these findings, we performed lung resections in 9 patients, who were judged contraindications for lung resection by the results of UPAO, and no patient suffered postoperative cardio-pulmonary complications. Total pulmonary vascular resistance immediately after the right middle and lower lobectomy were equal to those during SPAO. Thus we conclude that SPAO is very useful technique for evaluating cardiopulmonary function after right lower lobectomy or middle and lower lobectomy, and that it would be possible to extend the boundaries of safety for lung resection based on evaluation of cardio-pulmonary function by SPAO.


Advances in reducing the burden of vascular disease in type 2 diabetes.
1. The epidemic of diabetes is accelerating and the World Health Organization estimates that the number of people affected worldwide will grow from 171 million in 2000 to 366 million by 2030. 2. The main causes of death and disability in individuals with type 2 diabetes are macrovascular and microvascular disease, and blood pressure is one of the main determinants of vascular complications in this population. 3. While randomized trials have demonstrated that blood pressure lowering reduces vascular complications in subjects with type 2 diabetes and hypertension, ADVANCE was designed to determine whether the addition of a fixed combination of perindopril and indapamide, on top of comprehensive and effective cardiovascular treatments and glucose control therapy, would produce further benefits, irrespective of the initial blood pressure. 4. The blood pressure lowering arm of ADVANCE has demonstrated that the simple addition of the fixed combination of perindopril and indapamide compared to matching placebo, significantly reduces combined macrovascular and microvascular complications by 9%, all-cause mortality by 14% and cardiovascular death by 18%. It also reduces total coronary events by 14% and all renal events and microalbuminuria by 21%. 5. Similar benefits were observed in participant sub-groups characterized by age, sex, baseline blood pressure, previous vascular diseases and concomitant cardiovascular therapy including blood pressure lowering therapy. 6. Successful implementation of this treatment, with a single combination tablet of perindopril and indapamide, should be practical and affordable in most clinical settings worldwide and has the capacity to save countless lives and to reduce the burden of coronary disease and renal disease burden among millions of people with type 2 diabetes.


Association of drinking behaviors with TXNIP DNA methylation levels in leukocytes among the general Japanese population.
Background: Thioredoxin-interacting protein (TXNIP) controls the cellular redox balance by binding to and inhibiting the expression and function of thioredoxin. DNA methylation of the TXNIP gene is involved in the regulation of TXNIP mRNA expression. Changes in TXNIP DNA methylation levels are associated with the development of various diseases such as type 2 diabetes mellitus (T2DM). However, few studies have focused on the influence of lifestyle factors such as alcohol intake on TXNIP DNA methylation.Objectives: This research examines the association of drinking behaviors with TXNIP DNA methylation levels in the general Japanese population.Methods: We conducted a cross-sectional study of 404 subjects (176 males and 228 females) who were divided into non-, moderate and heavy drinkers based on self-reported drinking behaviors. TXNIP DNA methylation levels in leukocytes were determined using a pyrosequencing assay.Results: The mean TXNIP DNA methylation level in heavy drinkers (74.2%) was significantly lower than that in non- and moderate drinkers (non: 77.7%, p < .001; moderate: 76.6%, p = .011). Multivariable linear regression analysis showed that log-transformed values of daily (b = -1.34; p < .001) and cumulative (b = -1.06; p = .001) alcohol consumption were associated with decreased TXNIP DNA methylation levels.Conclusion: TXNIP DNA methylation levels in heavy drinkers was lower than in non- and- moderate drinkers. Decreased TXNIP DNA methylation level increases the expression of TXNIP and elevates the risk of developing of diseases such as T2DM. Therefore, decreasing alcohol use in heavy drinkers may lessen the likelihood of some alcohol-related illnesses moderated through TXNIP DNA methylation.


Purification and properties of a DNase inhibitor from Nicotiana tabacum cell cultures.
Extraction of Nicotiana tabacum cell cultures, chromatography on DEAE-cellulose and gel filtration resulted in a homogeneous protein (Mr = 14500), which strongly reduces the hydrolysis of Escherichia coli DNA by DNase I. DNA degradation by micrococcal nuclease is not inhibited. The inhibitor protein interacts with DNase I in the absence of DNA, as determined by the partial quenching of protein intrinsic fluorescence; a 1:1 stoichiometry is deduced. From the reduction of DNase I activity with increasing inhibitor concentration apparent equilibrium constants for the inhibitor X DNase-I complex have been calculated. This interaction is strongly temperature-dependent; at 20 degrees C and 26 degrees C dissociation constants of 5 nM and 110 nM, respectively, were determined. As a consequence a rather high enthalpy of interaction can be estimated.


Nur77 is differentially modified in PC12 cells upon membrane depolarization and growth factor treatment.
The rat pheochromocytoma cell line PC12 can be induced by growth factors to undergo proliferation and neuronal differentiation. These cells also have excitable membranes that can be depolarized by neurotransmitters or elevated levels of extracellular KCl. Treatment of PC12 cells with growth factors or membrane-depolarizing agents rapidly activates the expression of specific genes whose products are thought to mediate the subsequent biological responses. One such gene, nur77, is a member of the steroid and thyroid hormone receptor gene superfamily. We have identified the Nur77 protein and shown that it is synthesized rapidly and transiently in PC12 cells following stimulation, has a short half-life of 30 to 40 min, and is located in both the nucleus and the cytoplasm. Nur77 is posttranslationally modified, primarily by phosphorylation on serine residues. Phosphopeptide analysis reveals that Nur77 is modified differently upon membrane depolarization than after treatment with growth factors. We hypothesize that the activity of Nur77 is regulated by both differential gene expression and posttranslational modification and that these modes of regulation contribute to distinct downstream responses specific to membrane depolarization and growth factor treatment.


[The tongue mast cells of Bufo marinus L. toad: characterization of glycoconjugates with conventional and lectin methods].
Mast cell populations in mammals have been recognized as morphologically and functionally heterogeneous. In situ characterization of these cell type carbohydrates with conventional histochemical techniques and with lectin histochemistry has not been afforded in amphibian species. Different conventional staining methods for complex carbohydrates and 18 different, biotin or peroxidase conjugated, lectins were used in paraffin embedded Rossman fluid-fixed sections of mid-central region of the toad tongue. Conventional carbohydrate histochemistry showed a single type of mast cells with a variable concentration of highly sulphated glycosaminoglycans. Lectin histochemistry showed partial heterogeneity in the mast cell population. Most cells contain N-linked and O-linked oligosaccharides with variable quantities of Man but high of GlcNAc and terminal Gal-beta (1,3)-GaINAc residues. A considerable number of mast cells showed proteins or oligosaccharides with terminal sialic acid residues but only few mast cells contain terminal fucose. Discussion is made comparing these results with similar histochemical studies and with functional experimental studies performed in human and rat mast cells.


Role of corticotropin-releasing hormone in onset of labour.
Corticotropin-releasing hormone (CRH) derived from the placenta is secreted into the maternal circulation in large amounts during the third trimester of human pregnancy and may have an important role in the onset of labour. Although the biological role of CRH remains enigmatic, the presence of functional CRH receptors in the myometrium suggests that CRH may modulate myometrial contractility and hence parturition. CRH action is mediated via multiple receptor subtypes and pregnancy results in differential receptor expression. These receptors are primarily linked to the adenylate cyclase second messenger system, which would help the intracellular microenvironment to maintain the required myometrial quiescence. CRH can exert further actions such as inhibition of prostaglandin production to prevent contractions. At term under the influence of oxytocin there is a modification in the coupling mechanisms that leads to a decrease in the biological activity of the CRH receptor and in the generation of cyclic adenosine monophosphate which favours myometrial contractions. CRH, via distinct receptor subtypes, is then able to enhance the contractile response of the myometrium. This hypothesis places CRH in a central role in coordinating the smooth transition from a state of relaxation to one of contraction.


Oral melanoacanthoma with prominent spongiotic intraepithelial vesicles.
Oral melanoacanthoma is a rarely-reported, benign pigmented lesion of oral mucosa. Spongiosis was only recently mentioned as a light-microscopical finding occasionally seen in oral melanoacanthoma, and the formation of spongiotic intraepithelial vesicles to our knowledge has never been reported. The authors discuss a case of a 34-year-old, black woman who acutely developed extensive hyperpigmented oral lesions which on the biopsy showed both intraepithelial dendritic melanocytes and severe spongiosis forming intraepithelial vesicles. These lesions resolved spontaneously in a few months which led the authors to believe that the term melanoacanthoma, implying that it is a tumor, is a misnomer. This report supports findings of other investigators who consider oral melanoacanthoma to be a mucositis with unusual proliferation of dendritic melanocytes in the epidermis.


Gene expression patterns during the larval development of European sea bass (dicentrarchus labrax) by microarray analysis.
During the larval period, marine teleosts undergo very fast growth and dramatic changes in morphology, metabolism, and behavior to accomplish their metamorphosis into juvenile fish. Regulation of gene expression is widely thought to be a key mechanism underlying the management of the biological processes required for harmonious development over this phase of life. To provide an overall analysis of gene expression in the whole body during sea bass larval development, we monitored the expression of 6,626 distinct genes at 10 different points in time between 7 and 43 days post-hatching (dph) by using heterologous hybridization of a rainbow trout cDNA microarray. The differentially expressed genes (n = 485) could be grouped into two categories: genes that were generally up-expressed early, between 7 and 23 dph, and genes up-expressed between 25 and 43 dph. Interestingly, among the genes regulated during the larval period, those related to organogenesis, energy pathways, biosynthesis, and digestion were over-represented compared with total set of analyzed genes. We discuss the quantitative regulation of whole-body contents of these specific transcripts with regard to the ontogenesis and maturation of essential functions that take place over larval development. Our study is the first utilization of a transcriptomic approach in sea bass and reveals dynamic changes in gene expression patterns in relation to marine finfish larval development.


Effects of Changtai granules, a traditional compound Chinese medicine, on chronic trinitrobenzene sulfonic acid-induced colitis in rats.
To study the effects of Changtai granules (CTG), a traditional compound Chinese medicine, on chronic trinitrobenzene sulfonic acid-induced colitis in rats. Healthy adult Sprague-Dawley (SD) rats of both sexes, weighing 250-300 g, were employed in the present study. The rat colitis models were induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) enemas at a concentration of 100 mg/kg in 50% ethanol. The experimental animals were randomly divided into dexamethasone (DX) treatment, CTG treatment, and model control groups, which were intracolicly treated daily with DX (0.2 mg/kg), CTG at doses of 2.9, 5.7 and 11.4 g crude drug/kg, and the equal amount of saline respectively from 6 h following induction of the colitis in rats inflicted with TNBS to the end of study. A normal control group of rats treated without TNBS but saline enema was also included in the study. After 3 wk of treatment, the animals were assessed for colonal inflammatory and ulcerative responses with respect to mortality, frequency of diarrhea, histology and myeloperoxidase activity (MPO). The therapeutic effect of CTG on ulcerative colitis (UC) was better than DX. CTG effectively inhibited the activity of granulocytes, macrophages and monocytes in a dose-dependent manner. Also it reduced MPO and formation of inflammation in colonic mucosal tissue. Furthermore, administration of CTG significantly prevented body mass loss and death, and decreased frequency of diarrhea in UC rats, when compared with the model control group rats. CTG would prove to be an ideal drug for chronic UC, and is warranted to be studied further.


The effects of a tobacco intervention on binge drinking among African American light smokers.
Tobacco use and binge drinking are commonly associated with each other and research has shown that reducing smoking may influence alcohol use in alcohol-dependent populations. Although African Americans report a lower prevalence of binge drinking than other racial/ethnic groups, they are more likely to report consequences associated with this behavior. The aim of this article was to study the effect of a smoking intervention (counseling) on binge drinking prevalence and the frequency and average daily alcohol consumption in a sample of African American light smokers (those who smoke 10 cigarettes or less per day). Multivariate models were used to assess whether counseling type (health education or motivational interviewing) affected binge drinking prevalence and frequency or average daily alcohol consumption among participants (N = 755). Generalized linear models were run to assess the mediation effect of smoking cessation on the relationship between the counseling intervention and drinking outcomes. Finally, smoking cessation (regardless of counseling type) was assessed to determine the effect on alcohol use outcomes. Overall, counseling type was significantly related to a reduction in past 30-day binge drinking prevalence at week 8 among participants in the health education counseling (P = .045); however, these results diminished within 6 months. Smoking cessation did not appear to mediate the relationship between counseling type and alcohol use outcomes. Regardless of counseling type, individuals who quit smoking within the first 8 weeks of the study reported lower past 30-day binge drinking prevalence at week 8 than those who did not quit during the first 8 weeks (P = .035), but the effect was not sustained at the end of the study (week 26). These results show that tobacco interventions can affect binge drinking, but the effect does not appear to be sustained over time.


Prospective assessment of primary rectal cancer response to preoperative radiation and chemotherapy using 18-fluorodeoxyglucose positron emission tomography.
The purpose of this prospective study was to determine the ability of fluorine-18 fluorodeoxyglucose positron emission tomography to assess extent of pathologically confirmed rectal cancer response to preoperative radiation and 5-fluorouracil-based chemotherapy. Patients with primary rectal cancer deemed eligible for preoperative radiation and 5-fluorouracil-based chemotherapy because of a clinically bulky or tethered tumor or endorectal ultrasound evidence of T3 and/or N1 were prospectively enrolled. Positron emission tomography and CT scans were obtained before preoperative radiation and 5-fluorouracil-based chemotherapy (5,040 cGy to the pelvis and 2 cycles of bolus 5-fluorouracil with leucovorin) and repeated four to five weeks after completion of radiation and 5-fluorouracil-based chemotherapy. In addition to routine pathologic staging, detailed assessment of rectal cancer response to preoperative radiation and 5-fluorouracil-based chemotherapy was performed independently by two pathologists. Positron emission tomography parameters studied included conventional measures such as standardized uptake value (average and maximum), positron emission tomography-derived tumor volume (size), and two novel parameters: visual response score and change in total lesion glycolysis. Of 21 patients enrolled, prospective data (pretreatment and posttreatment positron emission tomography, and complete pathologic assessment) were available on 15 patients. All 15 demonstrated pathologic response to preoperative radiation and 5-fluorouracil-based chemotherapy. This was confirmed in 100 percent of the cases by positron emission tomography compared with 78 percent (7/9) by CT. In addition, one positron emission tomography parameter (visual response score) accurately estimated the extent of pathologic response in 60 percent (9/15) of cases compared with 22 percent (2/9) of cases with CT. This pilot study demonstrates that fluorine-18 fluorodeoxyglucose positron emission tomography imaging adds incremental information to the preoperative assessment of patients with rectal cancer. However, further studies in a larger series of patients are needed to verify these findings and to determine the value of fluorine-18 fluorodeoxyglucose positron emission tomography in a preoperative strategy aimed at identifying patients suitable for sphincter-preserving rectal cancer surgery.


In vitro cytotoxicity testing of three zinc metal salts using established fish cell lines.
The utilisation of fish cell lines has proven to be a valuable, rapid and cost-effective tool in the ecotoxicological assessment of chemicals and environmental samples. The main objective of this study was to investigate the value of multiple endpoint measurements in evaluating the cytotoxicity of three divalent zinc salts in three established fish cell lines (EPC, CHSE and RTG-2) and the potential for their employment as effective screening tools for zinc contaminated environmental samples. A significant stimulatory effect was detected with the neutral red assay in EPC and RTG-2 cells exposed to the lower doses of some zinc compounds. Significant (p < or = 0.01) lactate dehydrogenase release was detectable only with the highest exposure concentration of ZnCl2. Toxicity ranking based on IC50 values calculated from the neutral red and coomassie blue assay data found that in general, ZnC2 was the most cytotoxic metal compound to the cell lines employed. Differential cell sensitivities were observed to be dependant on the particular compound tested and the endpoint employed. It was found that the use of light microscopy in the identification of cell morphological changes was a valuable adjunct in verifying the results of colorimetric tests. In conclusion, careful consideration should be given to study design and statistics applied and use of a battery style approach is recommended for toxicological screening studies.


Health Inequities Among People Who Use Drugs in a Post-Dobbs America: The Case for a Syndemic Analysis.
Punitive policy responses to substance use and to abortion care constitute direct attacks on personal liberty and bodily autonomy. In this article, we leverage the concept of "syndemics" to anticipate how the already synergistic stigmas against people who use drugs and people who seek abortion services will be further compounded the Dobbs decision.


Effects of removal and reapplication of K(+) and Cl(-) on spontaneous electrical activity, slow wave, in the circular muscle of the guinea-pig gastric antrum.
In the circular muscle of guinea-pig gastric antrum, the effects of removal and reapplication of K(+) and Cl(-) were studied on the slow wave, which consists of the lower, first and upper, second components. The first component appeared to be triggered by the driving potential generated in the interstitial cells. K(+) removal slightly depolarized the membrane, increased frequency, and shortened the first component and driving potential, and K(+) reapplication hyperpolarized and prolonged these potentials transiently. Ouabain abolished the K(+)-induced hyper-polarization but had no inhibitory effect on the K(+)-induced potentiation. The K(+)-induced prolongation was much reduced in Ca(2+)-deficient and increased in Ca(2+)-excess solution. BAPTA-AM, thapsigargin, and cyclopiazonic acid shortened the slow wave and inhibited the K(+)-induced prolongation but did not block the slow wave. Effects of Cl(-) removal were stronger than K(+) removal in shortening and increasing the frequency. In Cl(-)-deficient solution, no prolongation was observed on K(+) reapplication. Although no conclusive evidence was obtained as to the ionic mechanism involved in the effects of K(+) or Cl(-) removal and reapplication, a possibility is considered that the sarcoplasmic reticulum is involved in determining the duration of the driving potential and the first component of the slow wave.


TOR and PKA pathways synergize at the level of the Ste11 transcription factor to prevent mating and meiosis in fission yeast.
In the fission yeast Schizosaccharomyces pombe, the TOR (target of rapamycin) and PKA (protein kinase A) signaling transduction pathways regulate the expression of genes required for cell growth and sexual differentiation in response to the nutritional environment. Inhibition of Tor2 signaling results in the induction of genes involved in sexual differentiation, and the cells undergo mating and meiosis, even under good nutritional conditions. The same phenotype is observed in mutants in which the PKA pathway is inactive. By contrast, Tor2 overexpression or mutations that hyperactivate PKA signaling impair sexual differentiation, even under poor nutritional conditions. Accordingly, a very important question is to understand the molecular mechanism by which these two pathways coordinately regulate gene expression in response to nutrients. Here we demonstrate that TOR and PKA pathways operate coordinately to negatively regulate sexual differentiation by inhibiting the nuclear accumulation of the Ste11 transcription factor. However, the Tor2 pathway is unable to block the nuclear localization of Ste11 under good nutritional conditions when the PKA pathway is inactive. Using microarray analyses, we found that both pathways inhibit sexual differentiation by blocking ste11-dependent gene expression. We conclude that both the PKA and the TOR pathways inhibit Ste11 nuclear accumulation to repress Ste11-dependent gene expression. However, the PKA pathway plays a quantitatively more important role than the TOR pathway in this process.


Effectiveness of Platelet-Rich Plasma for COVID-19-Related Olfactory Dysfunction: A Controlled Study.
To investigate the effectiveness of platelet-rich plasma (PRP) injection into the olfactory clefts of coronavirus disease 2019 (COVID-19) patients with persistent olfactory dysfunction (OD). Controlled study. Multicenter study. From March 2022 to November 2022, COVID-19 patients with persistent OD were recruited from three European hospitals to undergo PRP injections into the olfactory clefts. Olfactory function was evaluated at baseline and 10 weeks postinjection with the Olfactory Disorder Questionnaire (ODQ) and threshold, discrimination, and identification (TDI) test. Data were compared with a control group of untreated patients. Eighty-one patients who underwent PRP injection and 78 controls were included. Sixty-five PRP patients (80.3%) experienced subjective smell improvement after a mean duration of 3.4 ± 1.9 weeks. The parosmia, life quality statement, and ODQ sub- and total scores significantly decreased from pre- to 10-week postinjection in the PRP group. The TDI sub- and total scores significantly increased 10 weeks postinjection. In controls, the ODQ score did not change over time, while the discrimination, identification, and total TDI scores significantly increase after 10 weeks of follow-up. The 10-week TDI and ODQ scores were significantly better in the PRP group compared with the controls. Patients who underwent PRP injection reported better 10-week subjective and objective smell outcomes than controls. Future randomized-controlled studies using saline injection into the olfactory cleft of controls are needed to determine the superiority of PRP over placebo.


Characterization of feline T cell receptor gamma (TCRG) variable region genes for the molecular diagnosis of feline intestinal T cell lymphoma.
A diagnosis of intestinal lymphoma is currently made on the basis of clinical and morphologic criteria. This can prove problematic for many reasons that include inadequate sample size, the coexistence of lymphoma and inflammation, and the inability to assess architectural integrity of all tissue compartments in biopsy specimens obtained endoscopically. The detection of a clonal population of cells in a lymphoproliferative lesion represents an important criterion for the diagnosis of neoplasia, but this has not been assessed in feline intestinal lymphoma. T cell receptor gamma (TCRG) gene rearrangement analysis using polymerase chain reaction (PCR) is a methodology that can be used to detect clonality in T cell populations. The basis of this assay depends on the assessment of the junctional diversity that results from rearrangement of TCRG V (variable) and J (joining) gene segments. Feline TCRG transcripts from normal small intestine and spleen were obtained using a rapid amplification of cDNA ends (5'RACE) method. Limited diversity of TCRG V and J gene segments was observed. The high degree of sequence homology in the TCRG V and J gene segments was exploited to develop a PCR test for the assessment of TCRG V--J junctional diversity and hence clonality determination of T cell populations in cats. Molecular clonality determination was applied to feline intestinal lymphoplasmacytic inflammatory bowel disease (IBD) (9 cats), and transmural and mucosal T cell lymphoma (28 cats). Clonal rearrangement of the TCRG V--J junction was detected in 22 of 28 intestinal T cell lymphomas, and oligoclonality was detected in 3 intestinal T cell lymphomas. This contrasted with the detection of polyclonal rearrangement in normal intestinal tissues (3 cats) and in lymphoplasmacytic IBD (9 cats). It is proposed that assessment of TCRG V--J junctional diversity for the detection of clonality represents an important adjunctive tool for the diagnosis of T cell lymphoma in the cat.


Studies on the size and shape of rabbit intestinal glucoamylase-maltase complex.
Rabbit intestinal glucoamylase-maltase was examined in detail with respect to its molecular weight, sedimentation, diffusion and viscosity. It is a large asymmetrical molecule, with a molecular weight of 750 000-760 000. Its appearance under the electron microscope supports the idea that it is a long string (62.0 nm) consisting of eight beads of diameter 6.0 nm each and a surface-to-surface interbead distance of approx. 2.0 nm. The shape of the enzyme derived from its hydrodynamic behaviour by using the string-of-spherical-beads model originally proposed by Kuhn [(1932) Z. Phys. Chem. Abt. A 161, 1-32] and later modified by Shulman [(1953) J. Am. Chem. Soc. 75, 5846-5852] fits moderately well with the electron-microscopic picture. The beads might represent about six subunits, and the absence of sulphur from the enzyme and the inability to dissociate the enzyme by conventional methods indicate the possibility of unusual covalent cross-linking between the subunits and between the beads.


Histological Grade and Magnetic Resonance Imaging Quantitative T1rho/T2 Mapping in Osteoarthritis of the Knee: A Study in 20 Patients.
BACKGROUND Magnetic resonance imaging (MRI) of osteoarthritis (OA) of the knee is a preoperative method of joint assessment. Histology of the joint is invasive and performed after surgery. T1rho/T2 MRI mapping is a new preoperative method of quantifying joint changes. This study aimed to analyze and compare the histological changes in the joint cartilage with the use of quantitative T1rho/T2 MRI mapping in patients with OA of the knee. MATERIAL AND METHODS Twenty patients with OA of the knee (20 knees) underwent preoperative MRI with T1rho mapping, T2 mapping, T1-weighted, and T2-weighted fat-suppressed MRI sequences. The degree of OA of the knee on MRI was graded according to the Osteoarthritis Research Society International (OARSI) criteria and the Kellgren-Lawrence grading system. Histological grading of OA used the OARSI criteria. Four tibiofemoral condyles were assessed histologically, and the degree of cartilage destruction was determined using the OARSI criteria. Two investigators performed cartilage segmentation for T1rho/T2 values. RESULTS Histology of the four knee joint condyles confirmed mild to severe OA. The histology of the cartilage thickness (P<0.001) and the MRI findings of the distal medial condyle (P<0.00) were significantly different from the other three knee joint condyles. The T2 and T1rho values of each condyle were significantly correlated with the histological grade (II-IV) of the joint condyles, including the cartilage volume, cartilage defects, thickness, and bone lesions (P<0.05). CONCLUSIONS In 20 patients with OA of the knee, preoperative T2/T1rho MRI identified Grade II-IV OA changes in the joint.


Differences in mechanical properties lead to anomalous phase separation in a model cell co-culture.
During the morphogenesis of tissues and tumors, cells often interact with neighbors with different mechanical properties, but the understanding of its role is lacking. We use active Brownian dynamics simulations to study a model co-culture consisting of two types of cells with the same size and self-propulsion speed, but different mechanical stiffness and cell-cell adhesion. As time evolves, the system phase separates out into clusters with distinct morphologies and transport properties for the two cell types. The density structure factors and the growth of cell clusters deviate from behavior characteristic of the phase separation in binary fluids. Our results capture emergent structure and motility previously observed in co-culture experiments and provide mechanistic insights into intercellular phase separation during development and disease.


Assigning N-glycosylation sites of glycoproteins using LC/MSMS in conjunction with endo-M/exoglycosidase mixture.
The assignment of protein glycosylation sites and their microheterogeneities are of biological importance, yet such characterization is still considered to be analytically very challenging. Several approaches have been recently developed to improve the characterization of glycosylation sites of proteins, including lectin and HILIC enrichment-based methods coupled to mass spectrometry. However, unequivocal assignment of protein glycosylation remains to be a daunting task, prompting continuous demands for the development of sensitive and cutting-edge analytical approaches. beta-N-Acetylglucosaminidase (endo-beta-GlcNAc-ases, Endo-M) is an endoglycosidase capable of hydrolyzing N,N'-diacetylchitobiose moiety in N-linked oligosaccharides bound to the asparagine amino acid residue in various glycoproteins. An attractive feature of this enzyme is its ability to cleave the N,N'-diacetylchitobiose moiety while leaving an N-acetylglucosamine residue bound to the protein. This enzyme is also known to be inactive in the presence of core fucose residue linked to the reducing-end N-acetylglucosamine residue (GlcNAc). Here, we describe an approach capitalizing on these features of Endo-M to (a) determine the glycosylation sites of proteins and the occupancy of these sites, and (b) determine the attachment sites of fucose residue containing N-glycans. The latter is important because of its biological implications. Tryptically digested glycoproteins, which were subjected to Endo-M treatment, were analyzed by LC-MS/MS. Systematic evaluation of the activity of Endo-M toward different glycan structures indicated a dependence of enzyme activity on the complexity of the glycan structures. Efficient release of N-glycans using Endo-M is only achieved through the inclusion of a battery of exoglycosidases to reduce the complexity of the attached glycans and subsequently prompt an effective enzymatic release. Upon Endo-M/exoglycosidase treatment of tryptically digested glycoproteins, glycosylated sites retain GlcNAc residue. The resulting peptides with GlcNAc residues attached to the glycosylation sites are easily assigned through LC-MS/MS analysis and subsequent database searching of the generated tandem MS of such entities. Comparing the LC-MS/MS results of the tryptic digest of glycoproteins treated with PNGase F and Endo-M/exoglycosidases allowed the assignment of core fucose residues to N-glycan reducing-ends. The detection of glycosylation sites only in the tryptic digest of PNGase F treated samples suggested core fucosylation of the attached N-glycans to such sites. This strategy was initially validated using model glycoproteins. It also proved to be useful in determining the glycosylation sites of blood serum glycoproteins.


Amino acid sequence motifs essential for P0-mediated suppression of RNA silencing in an isolate of potato leafroll virus from Inner Mongolia.
Polerovirus P0 suppressors of host gene silencing contain a consensus F-box-like motif with Leu/Pro (L/P) requirements for suppressor activity. The Inner Mongolian Potato leafroll virus (PLRV) P0 protein (P0(PL-IM)) has an unusual F-box-like motif that contains a Trp/Gly (W/G) sequence and an additional GW/WG-like motif (G139/W140/G141) that is lacking in other P0 proteins. We used Agrobacterium infiltration-mediated RNA silencing assays to establish that P0(PL-IM) has a strong suppressor activity. Mutagenesis experiments demonstrated that the P0(PL-IM) F-box-like motif encompasses amino acids 76-LPRHLHYECLEWGLLCG THP-95, and that the suppressor activity is abolished by L76A, W87A, or G88A substitution. The suppressor activity is also weakened substantially by mutations within the G139/W140/G141 region and is eliminated by a mutation (F220R) in a C-terminal conserved sequence of P0(PL-IM). As has been observed with other P0 proteins, P0(PL-IM) suppression is correlated with reduced accumulation of the host AGO1-silencing complex protein. However, P0(PL-IM) fails to bind SKP1, which functions in a proteasome pathway that may be involved in AGO1 degradation. These results suggest that P0(PL-IM) may suppress RNA silencing by using an alternative pathway to target AGO1 for degradation. Our results help improve our understanding of the molecular mechanisms involved in PLRV infection.


Who falls between the cracks? Identifying eligible PrEP users among people with Sub-Saharan African migration background living in Antwerp, Belgium.
This study produces an estimate of the proportion of eligible PrEP users among people of Sub-Saharan African background based on the Belgian PrEP eligibility criteria and examines associations with socio-economic and demographic characteristics. We performed logistic regression analysis on data of a representative community-based survey conducted among Sub-Saharan African communities (n = 685) living in Antwerp. Almost a third (30.3%) of the respondents were eligible to use PrEP. Those who were male, single, lower educated, undocumented, and had experienced forced sex were more likely to be eligible for PrEP use. The findings highlight the importance of taking intra-, interpersonal and structural HIV risk factors into account. The study shows high unmet PrEP needs in this population, especially among those with high vulnerability for HIV acquisition. A better understanding of barriers to PrEP use in this population group is needed to allow for equitable access.


Differential gene expression induced by insulin and insulin-like growth factor-II through the insulin receptor isoform A.
The human insulin receptor (IR) exists in two isoforms (IR-A and IR-B). IR-A is a short isoform, generated by the skipping of exon 11, a small exon encoding for 12 amino acid residues at the carboxyl terminus of the IR alpha-subunit. Recently, we found that IR-A is the predominant isoform in fetal tissues and malignant cells and binds with a high affinity not only insulin but also insulin-like growth factor-II (IGF-II). To investigate whether the activation of IR-A by the two ligands differentially activate post-receptor molecular mechanisms, we studied gene expression in response to IR-A activation by either insulin or IGF-II, using microarray technology. To avoid the interfering effect of the IGF-IR, IGF-II binding to the IR-A was studied in IGF-IR-deficient murine fibroblasts (R- cells) transfected with the human IR-A cDNA (R-/IR-A cells). Gene expression was studied at 0.5, 3, and 8 h. We found that 214 transcripts were similarly regulated by insulin and IGF-II, whereas 45 genes were differentially transcribed. Eighteen of these differentially regulated genes were responsive to only one of the two ligands (12 to insulin and 6 to IGF-II). Twenty-seven transcripts were regulated by both insulin and IGF-II, but a significant difference between the two ligands was present at least in one time point. Interestingly, IGF-II was a more potent and/or persistent regulator than insulin for these genes. Results were validated by measuring the expression of 12 genes by quantitative real-time reverse transcriptase-PCR. In conclusion, we show that insulin and IGF-II, acting via the same receptor, may differentially affect gene expression in cells. These studies provide a molecular basis for understanding some of the biological differences between the two ligands and may help to clarify the biological role of IR-A in embryonic/fetal growth and the selective biological advantage that malignant cells producing IGF-II may acquire via IR-A overexpression.


Synthesis and relaxometry of high-generation (G = 5, 7, 9, and 10) PAMAM dendrimer-DOTA-gadolinium chelates.
A series of high-generation (G) ethylenediamine-core polyamidoamine (PAMAM) dendrimers corresponding to G = 5, 7, 9, and 10 were conjugated with the bifunctional chelate 2-(4-isothiocyanatobenzyl)- 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetate (p-SCN-Bz-DOTA). Gadolinium (III) ion was added to the macromolecules, and the 1/T1 and 1/T2 NMRD profiles were measured at 3 degrees, 23 degrees, and 37 degrees C. The synthesis resulted in preparations that ranged from an average of 127 chelates and 96 Gd3+ ions per G = 5 dendrimer to an average of 3727 chelates and 1860 Gd3+ ions per G = 10 dendrimer. At 20 MHz and 23 degrees C, the 1/T1 ion relaxivity increased from 30 mM(-1) s(-1) for the G = 5 to 35 mM(-1) s(-1) for the G = 7 PAMAM dendrimer-DOTA-Gd, reaching a plateau at 36 mM(-1) s(-1) for the G = 9 and G = 10 dendrimers. A similar plateau was observed for 1/T2 with values of 36 mM(-1) s(-1) for G = 5, 42 mM(-1) s(-1) for G = 7, and 45 mM(-1) s(-1) for the G = 9 and G = 10 dendrimers. This "saturation" of ion relaxivity for high-generation dendrimers occurred over the entire frequency range studied. The 1/T1 and 1/T2 relaxivities decreased as the temperature decreased for each generation of dendrimer studied, implying that slow water exchange of bound water molecules with the bulk solvent limits the relaxivity. In such circumstances, increases in the rotational correlation time of the macromolecules associated with higher generations of dendrimer does not result in significant increases in the ion relaxivity. Although the ion relaxivity does not increase, the total molecular relaxivities increased from 2880 mM(-1) s(-1) to 66960 mM(-1) s(-1) for the G = 5 to the G = 10 dendrimer. The current findings are relevant for the design of high-generation dendrimer-based receptor agents.


EP2 and EP4 receptor antagonists: Impact on cytokine production and β2 -adrenergic receptor desensitization in human airway smooth muscle.
Prostaglandin E2 (PGE2 ) is a key prostanoid known to have both proinflammatory and anti-inflammatory impact in the context of chronic respiratory diseases. We hypothesize that these opposing effects may be the result of different prostanoid E (EP) receptor-mediated signaling pathways. In this study, we focus on two of the four EP receptors, EP2 and EP4 , as they are known to induce cyclic adenosine monophosphate (cAMP)-dependent signaling pathways. Using primary human airway smooth muscle (ASM) cells, we first focussed on the PGE2 -induced production of two cAMP-dependent proinflammatory mediators: interleukin 6 (IL-6) and cyclo-oxygenase 2 production. We show that PGE2 -induced IL-6 protein secretion occurs via an EP2 -mediated pathway, in a manner independent of receptor-mediated effects on messenger RNA (mRNA) expression and temporal activation kinetics of the transcription factor cAMP response element binding. Moreover, stimulation of ASM with PGE2 did not establish a positive, receptor-mediated, feedback loop, as mRNA expression for EP2 and EP4 receptors were not upregulated and receptor antagonists were without effect. Our studies revealed that the EP2 , but not the EP4 , receptor is responsible for β2 -adrenergic desensitization induced by PGE2 . We demonstrate that PGE2 -induced heterologous receptor desensitization responsible for tachyphylaxis to short- (salbutamol) or long- (formoterol) β2 -agonists (measured by cAMP release) can be reversed by the EP2 receptor antagonist PF-04418948. Importantly, this study highlights that inhibiting the EP2 receptor restores β2 -adrenergic receptor function in vitro and offers an attractive novel therapeutic target for treating infectious exacerbations in people suffering from chronic respiratory diseases in the future.


Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI-Application in Premanifest Huntington's Disease.
Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington's disease (HD), regional brain atrophy begins many years prior to the motor onset (during the "premanifest" period), but the spatiotemporal pattern of regional atrophy across the brain has not been fully characterized. Here we demonstrate an online cloud-computing platform, "MRICloud", which provides atlas-based whole-brain segmentation of T1-weighted images at multiple granularity levels, and thereby, enables us to access the regional features of brain anatomy. We then describe a regression model that detects statistically significant inflection points, at which regional brain atrophy starts to be noticeable, i.e. the "change-point", with respect to a disease progression index. We used the CAG-age product (CAP) score to index the disease progression in HD patients. Change-point analysis of the volumetric measurements from the segmentation pipeline, therefore, provides important information of the order and pattern of structural atrophy across the brain. The paper illustrates the use of these techniques on T1-weighted MRI data of premanifest HD subjects from a large multicenter PREDICT-HD study. This design potentially has wide applications in a range of neurodegenerative diseases to investigate the dynamic changes of brain anatomy.