Purification and characterization of normal and mutant forms of T4 endonuclease V.
Endonuclease V of bacteriophage T4 has been purified to physical homogeneity from T4D-infected Escherichia coli 1100. The enzyme, whose molecular weight is 16,000, possessed two distinct catalytic activities, a pyrimidine dimer-DNA glycosylase and an apurinic/apyrimidinic endonuclease. They acted on UV-irradiated poly(dA) . poly(dT) in a sequential manner; the glycosylase cleaved the N-glycosyl bond between the 5'-pyrimidine of a dimer and the corresponding sugar and then the endonuclease hydrolyzed a phosphodiester bond on the 3'-side of the apyrimidinic site. The 5'-termini thus generated were phosphorylated by T4 polynucleotide kinase only after they had been subjected to direct photoreversal and then treated with alkaline phosphatase. By using two phage mutants, uvs-5 and uvs-13, it was shown that occurrence of an amber mutation in the denV gene caused a simultaneous loss of the two activities. Suppression of the mutation of uvs-5 rendered both activities partially active. When the mutation of uvs-13 was suppressed, a mutant form of enzyme that possessed only a glycosylase activity was produced. This suggests that there are two distinct domains in a single enzyme, each of which corresponds to one of the activities.


Establishing evidence-based physical activity guidelines: methods for the Study of Health and Activity in People with Spinal Cord Injury (SHAPE SCI).
Prospective, observational cohort study. This paper describes the rationale and methodology for the Study of Health and Activity in People with Spinal Cord Injury (SHAPE SCI). The study aims to (1) describe physical activity levels of people with different injury levels and completeness, (2) examine the relationship between physical activity, risk and/or presence of secondary health complications and risk of chronic disease, and (3) identify determinants of physical activity in the SCI population. Ontario, Canada. Seven hundred and twenty men and women who have incurred a traumatic SCI complete self-report measures of physical activity, physical activity determinants, secondary health problems and subjective well-being during a telephone interview. A representative subsample (n=81) participate in chronic disease risk factor testing for obesity, insulin resistance and coronary heart disease. Measures are taken at baseline, 6 and 18 months. SHAPE SCI will provide much-needed epidemiological information on physical activity patterns, determinants and health in people with SCI. This information will provide a foundation for the establishment of evidence-based physical activity guidelines and interventions tailored to the SCI community.


Correlation between virulence markers of Helicobacter pylori in the oral cavity and gastric biopsies.
The clinical outcome of Helicobacter pylori infection has been associated with virulence factors. The presence of these factors is useful as molecular markers in the identification of the high risk for developing severe gastric pathologies. To correlate the presence of virulence markers cagA and bab2A of H. pylori in oral and gastric biopsy samples. An observational, prospective, descriptive, and cross-sectional study was carried out between September 2011 and September 2012. Patients suffering dyspepsia with indication for upper gastrointestinal video endoscopy who attended the Gastroenterology Service of the Hospital Dr. Julio C. Perrando were included. Epidemiological investigation was completed. To detect the bacteria and their virulence genes, samples of saliva, dental plaque and gastric biopsy were taken and processed by PCR. Sixty-one patients were selected for this study (30 women and 31 men). H. pylori was detected in 31 gastric biopsies and 31 oral samples. Significant difference between oral and gastric samples was found in cagA genotype. Agreement between oral and gastric genotypes was found in 38.7% of samples from the same patient. This study is the first in provide information about the genotypes of the Argentinean Northeast H. pylori strains. Despite the high prevalence of H. pylori infection, the most of patients had less virulent genotypes in oral cavity and gastric tissue. The cagA / babA2 combination was not frequent in the samples studied. There was not a statistical correlation between the virulence genes and gastroduodenal or oral diseases. Although in some patients the same genotype was found both in oral and gastric samples, it cannot be ensure that they corresponding to the same strain because a DNA sequencing was not performed.


Technoeconomic Analysis of Negative Emissions Bioenergy with Carbon Capture and Storage through Pyrolysis and Bioenergy District Heating Infrastructure.
Bioenergy with carbon capture and storage (BECCS) has been identified as a cost-effective negative emission technology that will be necessary to limit global warming to 1.5 °C targets. However, the study of BECCS deployment has mainly focused on large-scale, centralized facilities and geologic sequestration. In this study, we perform technoeconomic analysis of BECCS through pyrolysis technology within a district heating system using locally grown switchgrass. The analysis is based on a unique case study of an existing switchgrass-fueled district heating system in the rural southeastern United States and combines empirical daily energy data with a retrospective analysis of add-on pyrolysis technology with biochar storage. We show that at current heating oil and switchgrass prices, pyrolysis-bioenergy (PyBE) and pyrolysis BECCS (PyBECCS) can each reach economic parity with a fossil fuel-based system when the prices of carbon is $116/Mg CO2-eq and $51/Mg CO2-eq, respectively. In addition, each can reach parity with a direct combustion bioenergy (BE) system when the prices of carbon is $264/Mg CO2-eq and $212/Mg CO2-eq, respectively. However, PyBECCS cannot reach economic parity with BE without revenue from carbon sequestration, while PyBE can, and in some cases, PyBECCS could counterintuitively require more reliance on fossil fuels than both the PyBE case and BE.


Effects of the glucagon-like peptide-1 receptor agonist liraglutide on 24-h ambulatory blood pressure in patients with type 2 diabetes and stable coronary artery disease: a randomized, double-blind, placebo-controlled, crossover study.
The glucagon-like peptide-1 receptor agonist liraglutide has been shown to reduce blood pressure (BP) in clinical trials using office BP measurements. However, the effects of liraglutide on 24-h BP and on the diurnal variation in BP have not been explored sufficiently. Forty-one patients with type 2 diabetes and stable coronary artery disease were randomized to receive liraglutide or placebo to a backbone therapy of metformin in this double-blind, placebo-controlled 12 along with 12 weeks crossover study. Ambulatory blood pressure monitoring (ABPM) was performed at the start and end of each intervention. Twenty-four individuals completed all 24-h BP measurements. Liraglutide, when compared with placebo, did not induce any significant changes in mean 24-h SBP [difference +1.8 mmHg (95% confidence interval, 95% CI: -4.33 to 7.93)] or DBP [+4.2 mmHg (-0.74 to 9.17)]. Twenty-four-hour BP profiles revealed a trend for increase in evening SBP and DBP [+9.2 mmHg (95% CI: 1.1-17.2) and +9.7 mmHg (95% CI: 3.9-15.5), respectively]. Mean heart rate significantly increased after liraglutide [+7.6 bpm (95% CI: 2.56-12.62)]. Liraglutide did not affect the BP variability or the nocturnal BP dipping. We could not demonstrate any BP-lowering effect of liraglutide when using 24-h ABPM. Liraglutide exhibited diurnal variation in the effect on BP without affecting the BP variability or nocturnal BP dipping.


[Primary carcinoma of Bartholin's glands with HPV 18 detection].
A primary carcinoma of the Bartholin's gland is a rarely type of vulva-carcinoma. The patients median age are 55 years and the etiology of the carcinoma are still unknown. Viral influences are discussed. Most frequent types of carcinomas of Bartholin's gland are those of squamous cells and adenocarcinomas. This article reports on a 55 year old patient with a primary carcinoma of Bartholin's gland. The human papillomavirus 18 was detected by a polymerase chain reaction. The correct diagnosis was delayed because of an initial misinterpretation of the carcinoma as a Bartholin cyst, marsupialization and bad healing of the wound. The patient underwent a radical vulvectomy with bilateral inguinofemoral lymphonodectomy in combination with radiation therapy.


Agreement between careHPV and hybrid capture 2 in detecting high-risk HPV in women in Tanzania.
Visual inspection of the cervix with acetic acid is used to control the burden of cervical cancer in low- and middle-income countries. This method has some limitations and HPV DNA testing may be an alternative, but it is expensive and requires a laboratory setup. Cheaper and faster HPV tests have been developed. This study describe the agreement between a fast HPV test (careHPV) and hybrid capture 2 (HC2) in detection of high-risk HPV among Tanzanian women. The study involved women attending routine cervical cancer screening at the Ocean Road Cancer Institute and Kilimanjaro Christian Medical Centre in Tanzania. The women were offered HIV testing. Two cervical samples were subsequently obtained; the first sample was processed at the clinics using careHPV and the second sample was transported to Denmark and Germany for cytology and HC2 analysis. Kappa statistic was calculated to assess the agreement between careHPV and HC2. The sensitivity, specificity and predictive values of careHPV were calculated using HC2 as reference. The analyses were done for the overall study population and stratified by testing site and HIV status. A total of 4080 women were enrolled, with 437 being excluded due to invalid information, lack of careHPV or HC2 results. Overall agreement between the tests was substantial with a kappa value of 0.69 (95% confidence interval [CI] 0.66-0.72). The sensitivity and specificity of careHPV was 90.7% (95% CI 89.6-91.8) and 84.2% (95% CI 81.2-86.8), respectively. The agreement was similar in the stratified analyses where the kappa values were 0.75 (95% CI 0.70-0.79) in women aged 25-34, 0.66 (95% CI 0.62-0.70) in women aged 35-60, 0.73 (95% CI 0.70-0.77) at the Ocean Road Cancer Institute, 0.64 (95% CI 0.60-0.69) at the Kilimanjaro Christian Medical Center, 0.73 (95% CI 0.68-0.79) in HIV-positive and 0.66 (95% CI 0.63-0.70) in HIV-negative women. The kappa value of 0.64 (95% CI 0.39-0.88) for cervical high-grade lesions indicates a substantial agreement between careHPV and HC2 in detecting HPV among women with cervical high-grade lesions. A substantial agreement was found between careHPV and HC2 in detecting HPV overall as well as detecting HPV among women with cervical high-grade lesions. However, given the limited resources available in low and middle-income countries, the HPV testing assay should be weighed against the cost-effectiveness of the test.


Simian virus 40-transformed human cells that express large T antigens defective for viral DNA replication.
Many types of human cells cultured in vitro are generally semipermissive for simian virus 40 (SV40) replication. Consequently, subpopulations of stably transformed human cells often carry free viral DNA, which is presumed to arise via spontaneous excision from an integrated DNA template. Stably transformed human cell lines that do not have detectable free DNA are therefore likely to harbor harbor mutant viral genomes incapable of excision and replication, or these cells may synthesize variant cellular proteins necessary for viral replication. We examined four such cell lines and conclude that for the three lines SV80, GM638, and GM639, the cells did indeed harbor spontaneous T-antigen mutants. For the SV80 line, marker rescue (determined by a plaque assay) and DNA sequence analysis of cloned DNA showed that a single point mutation converting serine 147 to asparagine was the cause of the mutation. Similarly, a point mutation converting leucine 457 to methionine for the GM638 mutant T allele was found. Moreover, the SV80 line maintained its permissivity for SV40 DNA replication but did not complement the SV40 tsA209 mutant at its nonpermissive temperature. The cloned SV80 T-antigen allele, though replication incompetent, maintained its ability to transform rodent cells at wild-type efficiencies. A compilation of spontaneously occurring SV40 mutations which cannot replicate but can transform shows that these mutations tend to cluster in two regions of the T-antigen gene, one ascribed to the site-specific DNA-binding ability of the protein, and the other to the ATPase activity which is linked to its helicase activity.


Update on the Role of the Non-Canonical Wnt/Planar Cell Polarity Pathway in Neural Tube Defects.
Neural tube defects (NTDs), including spina bifida and anencephaly, represent the most severe and common malformations of the central nervous system affecting 0.7-3 per 1000 live births. They result from the failure of neural tube closure during the first few weeks of pregnancy. They have a complex etiology that implicate a large number of genetic and environmental factors that remain largely undetermined. Extensive studies in vertebrate models have strongly implicated the non-canonical Wnt/planar cell polarity (PCP) signaling pathway in the pathogenesis of NTDs. The defects in this pathway lead to a defective convergent extension that is a major morphogenetic process essential for neural tube elongation and subsequent closure. A large number of genetic studies in human NTDs have demonstrated an important role of PCP signaling in their etiology. However, the relative contribution of this pathway to this complex etiology awaits a better picture of the complete genetic architecture of these defects. The emergence of new genome technologies and bioinformatics pipelines, complemented with the powerful tool of animal models for variant interpretation as well as significant collaborative efforts, will help to dissect the complex genetics of NTDs. The ultimate goal is to develop better preventive and counseling strategies for families affected by these devastating conditions.


Predictors of esophageal stricture formation post endoscopic mucosal resection.
Stricture formation is a common complication after endoscopic mucosal resection. Predictors of stricture formation have not been well studied. We conducted a retrospective, observational, descriptive study by using a prospective endoscopic mucosal resection database in a tertiary referral center. For each patient, we extracted the age, sex, lesion size, use of ablative therapy, and detection of esophageal strictures. The primary outcome was the presence of esophageal stricture at follow-up. Multivariate logistic regression was used to analyze the association between the primary outcome and predictors. Of 136 patients, 27% (n=37) had esophageal strictures. Thirty-two percent (n=44) needed endoscopic dilation to relieve dysphagia (median, 2; range, 1 to 8). Multivariate logistic regression analysis showed that the size of the lesion excised is associated with increased odds of having a stricture (odds ratio, 1.6; 95% confidence interval, 1.1 to 2.3; p=0.01), when controlling for age, sex, and ablative modalities. Similarly, the number of lesions removed in the index procedure was associated with increased odds of developing a stricture (odds ratio, 2.3; 95% confidence interval, 1.3 to 4.2; p=0.007). Stricture formation after esophageal endoscopic mucosal resection is common. Risk factors for stricture formation include large mucosal resections and the resection of multiple lesions on the initial procedure.


Serious Games for Sexual Health.
Program developers and researchers in the sexual health domain have increasingly embraced technological trends as they emerge. With the emergence of serious game applications to impact health behaviors, a natural step for research enquiry will be the investigation of serious games for sexual health education. We invited a panel of sexual health researchers who are working at the intersection of sexual health behavior change and technology applications to comment on the place of serious games in furthering the field of sexual health. The panel grappled with six questions.


Potential of endogenous regenerative technology for in situ regenerative medicine.
Endogenous regenerative technology (Endoret) involves the use of patient's own biologically active proteins, growth factors and biomaterial scaffolds for therapeutic purposes. This technology provides a new approach for the stimulation and acceleration of tissue healing and bone regeneration. The versatility and biocompatibility of using patient-derived fibrin scaffold as an autologous, biocompatible and biodegradable drug delivery system open the door to a personalized medicine that is currently being used in numerous medical and scientific fields including dentistry, oral implantology, orthopaedics, ulcer treatment, sports medicine and tissue engineering among others. This review discusses the state of the art and new directions in the use of endogenous technology in the repair and regeneration of injured tissues by means of a controlled and local protein and growth factor delivery. The next generations of engineering strategies together with some of the most interesting therapeutic applications are discussed together with the future challenges in the field.


Effects of adriamycin on lipid polymorphism in cardiolipin-containing model and mitochondrial membranes.
The effects of the anti-tumor drug adriamycin on lipid polymorphism in cardiolipin-containing model membranes and in isolated inner mitochondrial membranes has been examined by 31P-NMR. Adriamycin binding does not affect the macroscopic structure or local order in the phosphate region of cardiolipin liposomes. In cardiolipin liposomes and in cardiolipin-phosphatidylcholine (1:1) liposomes, the drug inhibits the ability of Ca2+ to induce the hexagonal HII phase. Adriamycin interaction with both dioleoylphosphatidylethanolamine-cardiolipin (2:1) and dioleoylphosphatidylethanolamine-phosphatidylserine (1:1) liposomes results in structural phase separation into a liquid-crystalline hexagonal HII phase for the phosphatidylethanolamine and a liquid-crystalline lamellar phase for the negatively charged phospholipid. Combined high-resolution 31P-NMR, electron microscopy and light scattering studies reveal the prominent fusion capacity of adriamycin towards cardiolipin-phosphatidylcholine small unilamellar vesicles. Addition of Ca2+ to total rat liver inner mitochondrial membrane lipids, dispersed in excess buffer, results in hexagonal HII formation for part of the phospholipids. By contrast, the original bilayer structure is completely conserved when the above experiment is performed in the presence of adriamycin. 31P-NMR spectra of isolated inner mitochondrial membranes are indicative of a bilayer organization for the majority of the phospholipids. Approximately 15% of the signal intensity originates from phospholipids which experience isotropic motion. Adriamycin addition almost completely eliminates the latter spectral component. In the absence of adriamycin, Ca2+ addition greatly increases the percentage of the phospholipids giving rise to an isotropic signal possibly indicating the formation of non-lamellar lipid structures. Adriamycin which specifically binds to cardiolipin (K. Nicolay et al. (1984) Biochim. Biophys. Acta 778, 359-371) completely blocks the Ca2+-induced structural reorganization of the lipids in this membrane.


Characterization of a polymer film optical fiber hydrophone for use in the range 1 to 20 MHz: a comparison with PVDF needle and membrane hydrophones.
A small aperture wideband ultrasonic optical fiber hydrophone is described. The transduction mechanism is based on the detection of acoustically induced changes in the optical thickness of a 25-microm thick parylene polymer film acting as a low finesse Fabry Perot (FP) interferometer that is deposited directly onto the end of a single mode optical fiber. The acoustic performance compares favorably with that of PVDF needle and membrane hydrophones with a peak noise-equivalent-pressure (without signal averaging) of 10 kPa over a 25-MHz measurement bandwidth, a wideband response to 20 MHz, and a near omnidirectional performance at 10 MHz. The dynamic range was 60 dB with an upper limit of linear detection of 11 MPa and a temporal stability of <5% over a period of 20 h. The hydrophone can also measure temperature changes with a resolution of 0.065 degrees C, offering the prospect of making simultaneous acoustic pressure and temperature measurements. The transduction parameters of the FP sensing element were measured, yielding an ultrasonic acoustic phase sensitivity of 0.075 rad/MPa and a temperature phase sensitivity of 0.077 rad/ degrees C. The ability to achieve high acoustic sensitivity with small element sizes and to repeatably fabricate rugged sensor downleads using polymer deposition techniques suggests that this type of hydrophone can provide a practical alternative to piezoelectric hydrophone technology.


Mean glucose values predict trauma patient mortality.
Tight glycemic control in a mixed surgical intensive care unit patient population has been associated with improved survival. We postulated targeted therapy to achieve glucose levels <140 mg/dL would reduce infectious complications and mortality in trauma patients admitted to the intensive care unit (ICU). Adult trauma patients admitted to our American College of Surgeons Level I Trauma Center ICU from July 2004 through June 30, 2006 were studied. Insulin therapy was instituted for ICU patients admitted after July 1, 2005 with glucose >140 mg/dL. Data on infections and all glucose values were collected. Multivariate analysis adjusting for age, Injury Severity Score, Glasgow Coma Scale Score, admit blood pressure, and intubation status was performed. Five thirty-one ICU patients were admitted with a mean Injury Severity Score of 23 +/- 13 and mean age of 45 years +/- 19 years. The admission, mean, and maximum glucoses were 141, 129, and 192 respectively. In multivariate analyses, increases in all three glucose values were associated with a significantly higher mortality, with the best model achieved using mean glucose with a receiver operating curve of 0.90. For mean glucose categories of >200 mg/dL, 141 mg/dL to 200 mg/dL, and </=140 mg/dL, the mortality was 40%, 20%, and 3.3%, respectively. Higher glucose levels were not associated with increased rates of infection after risk adjustment. After July 1, 2005, the use of insulin drips rose from 13% to 22% (p = 0.01), and the number of glucose checks per patient in the ICU rose from 27 to 43 (p < 0.02), and the percent of ICU patients with all glucose values less than 140 mg/dL rose from 59% to 78%. Higher glucose levels were significantly associated with increased risk of fatal outcome in trauma patients. Hyperglycemia was not an independent predictor of infectious complications. Despite the increased use of insulin drips and the higher number of glucose checks after adopting a stricter insulin treatment protocol, ICU outcomes remained unchanged.


Pilot of group intervention for bipolar disorder.
Objective. This pilot study aimed to determine whether a group based psychosocial intervention reduced rates of relapse, improved function and quality of life in people with bipolar disorder. Method. Patients with a diagnosis of bipolar disorder, types I and II were recruited in the Geelong Region of Victoria. Patients were assessed at baseline for psychiatric status, mood episode, function, and medication adherence. They were randomly assigned to either the intervention arm, a 12-week, structured group-based therapy as an adjunct to treatment as usual or the control arm, which consisted of treatment as usual, plus weekly phone calls. Participants were then followed up for a period of 3 months and assessed by a researcher blinded to treatment and control interventions. Results. Functioning as measured by the Global Assessment of Functioning (GAF) was significantly improved in the intervention group (P=0.008). The social relationships subscale on the (WHOQoL-BREF) showed significant results (P<0.05 level). There was also a positive trend in reduction of relapses in the intervention group. Conclusion. The use of a group intervention for bipolar disorder as an adjunct to usual treatment has potential benefits, both in reduction of relapse and improvement in functionality, and may be a cost effective way of delivering psychosocial treatments.


Mycobacterium behavior in wastewater treatment plant, a bacterial model distinct from Escherichia coli and Enterococci.
Mycobacteria are waterborne emerging pathogens causing infections in human. Mycobacteria have been previously isolated from wastewater and sludge, but their densities were not estimated due to cultural biases. In order to evaluate the impact of wastewater treatment processes on mycobacteria removal, we used a real time PCR method. First we compared six DNA extraction methods and second we used the more efficient DNA extraction procedure (i.e., enzymatic lysis combined with hexadecyltrimethylammonium bromide-NaCl procedure) in order to quantify Mycobacterium. With the aim to identify parameters that could serve as indicator of mycobacterial behavior, mycobacterial densities were measured in parallel to those of Escherichia coli and enterococci, and to concentrations of chemical parameters usually monitored in wastewater. Mycobacterium reached 5.5 × 10⁵ ± 3.9 × 10⁵ copies/L in the influent, but was not detected in the effluent after decantation and biofiltration. Most mycobacteria (98.6 ± 2.7%, i.e. 2.4 ± 0.7 log₁₀) were removed by the physical-chemical decantation, and the remaining mycobacteria were removed by biofiltration. In contrast, enterococci and E. coli were lightly removed by decantation step and mainly removed by biofiltration. Our results showed that Mycobacterium corresponds to a hydrophobic behavior linked to insoluble compound removal, whereas enterococci and E. coli refer to hydrophilic behaviors linked to soluble compound removals.


[Bacterial outer membrane vesicles as nano carriers to study immunological activities].
Objective: To prepare a nano-carrier based on combining bacterial outer membrane vesicles (OMV) with three block polymer pluronic F127 (PEO100-PPO65-PEO100) (OMV-F127) and to investigate its immunological activity. Methods: Attenuated salmonella (sal) was cultivated. OMV were separated by centrifugal ultrafiltration or ultrasonication, and OMV-F127 was prepared by mechanical extrudation method. The protein contents and compositions were tested with BCA and SDS-PAGE; the morphology of OMV, F127 and OMV-F127 were observed with FM and TEM; the particle sizes and their zeta potential were determined with DLS. Mouse macrophage RAW246.7 cells were treated with OMV-F127 (50 μg/mL, 100 μg/mL) in vitro, and the concentrations of IL-12, TNF-α and IFN-γ in culture supernatant were measured with ELISA kits. Results: The contents of protein in separated OMV by centrifugal ultrafiltration and ultrasonication were 2.8 mg/mL and 2.7 mg/mL, respectively. SDS-PAGE showed the marker protein OmpF/C in OMV. Under the FM and TEM, ball-like structure of F127 and OMV-F127 was observed. Size analysis revealed that the diameters of OMV, F127 and OMV-F127 were 72±2 nm, 90±3 nm and 92±2 nm, respectively. ELISA tests revealed that OMV-F127 significantly stimulated the secretion of IL-12, TNF-α and IFN-γ in RAW246.7 cells. Conclusion: A nano-carrier based on bacterial outer membrane vesicles has been prepared, which can stimulate the secretion of cytokines and may have immunomodulatory effects.


Active Ca2+ transport in primary cultures of rabbit kidney CCD: stimulation by 1,25-dihydroxyvitamin D3 and PTH.
Rabbit connecting tubules and cortical collecting ducts, which represent 79 +/- 5% of the calbindin-D28k-containing kidney cells, were isolated by immunodissection from the rabbit kidney superficial cortex and seeded on permeable filters. After 6 days in culture the monolayers developed a potential difference (PD) of -24 +/- 3 mV (lumen negative) and a transepithelial resistance (R) of 284 +/- 19 omega.cm2. Addition of 10(-6) M amiloride to or removal of Na+ from the mucosal side reversed the PD to +6 +/- 4 mV and concomitantly increased R to 660 +/- 122 omega.cm2. The cells developed functional parathyroid hormone (PTH) and arginine vasopressin receptors, but calcitonin receptors were absent. The monolayer actively absorbed Ca2+ against an electrochemical gradient with a rate of 121 +/- 13 nmol.h-1.cm-2. Removal of serosal Na+ inhibited Ca2+ absorption by 63 +/- 8%. Exposure to 10(-7) M 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] for 48 h or 10(-7) M bovine PTH (bPTH)-(1-34) for 1 h, increased transcellular Ca2+ absorption by 53 +/- 13% or 24 +/- 8%, respectively. The effects of 1,25(OH)2D3 and PTH in combination were neither additive nor potentiating. In addition, the cultured cells expressed calbindin-D28k, and, after exposure to 10(-7) M 1,25(OH)2D3 for 48 h, the calbindin-D28k content increased fourfold.


Vertigo and vestibular rehabilitation.
The role of rehabilitation in the management of vertigo is limited to a very specific group of conditions. An Occupational therapist who is a part of the multidisciplinary team treating the vertiginous patient, with the knowledge of physiology and therapeutic benefit of vestibular rehabilitation can widen the rehabilitation spectrum for various diseases producing vertigo and dysequilibrium, to resolve or minimise these symptoms. The present article reviews the need for vestibular rehabilitation and the different conditions needing the same along with its characteristics, physiology and various exercises prescribed.


[Accumulation of lipid peroxidation products in the human lens during cataract maturation].
130 human lens impaired with cataract of various etiology (senile, traumatic, complicated cataract) and of different cataract maturation (initial, immature, quite mature, mature, over-maturated) as well as 18 transparent lens were studied. Primary products of lipid peroxidation (LP, hydroperoxides), secondary products (ketodiens) and the end products (Shiff bases) were distinctly accumulated in cataract lens. Intermediates of primary LP products were accumulated in lens within early periods of cataract development but the end LP products were mainly found at the late steps of cataract formation. Accumulation of the LP products was not distinctly different in cataracts with various clinical manifestations but similar in the level of maturation, thus indicating that lipid peroxidation plays a universal role in the process of lens opacity. The data obtained suggest that inhibitors of free radical oxidation (antioxidants) may be employed in medicamental treatment of cataracts.


Peptidoglycan Deacetylases in Bacterial Cell Wall Remodeling and Pathogenesis.
The bacterial cell wall peptidoglycan (PG) is a dynamic structure that is constantly synthesized, re-modeled and degraded during bacterial division and growth. Postsynthetic modifications modulate the action of endogenous autolysis during PG lysis and remodeling for growth and sporulation, but also they are a mechanism used by pathogenic bacteria to evade the host innate immune system. Modifications of the glycan backbone are limited to the C-2 amine and C-6 hydroxyl moieties of either GlcNAc or MurNAc residues. This paper reviews the functional roles and properties of peptidoglycan de-Nacetylases (distinct PG GlcNAc and MurNAc deacetylases) and recent progress through genetic studies and biochemical characterization to elucidate their mechanism of action, 3D structures, substrate specificities and biological functions. Since they are virulence factors in pathogenic bacteria, peptidoglycan deacetylases are potential targets for the design of novel antimicrobial agents.


Education, Activity Engagement, and Cognitive Function in US Chinese Older Adults.
To examine whether and how early-life experiences such as years of schooling affect late-life cognitive function through a pathway of activity engagement. Prospective. We used data from 2 waves of the Population Study of Chinese Elderly in Chicago (PINE). PINE is the largest population-based epidemiological study of Chinese-American adults aged 60 and older in the greater Chicago area. Wave 1 data were collected for 2 years, from July 2011 to June 2013, and Wave 2 data were collected from 2013 to 2015; total sample size was 2,713. Education was measured in years of schooling. Activity engagement was assessed using 15 items grouped into two clusters: cognitive activity and social activity. Cognitive function was evaluated using five instruments to assess general mental status (Chinese Mini-Mental State Examination (C-MMSE)), episodic memory, perceptual speed, working memory, global cognition score. Adjusting for sociodemographic and health-related control variables, education measured at Wave 1 was associated with better global cognition (b = 0.025, p < .001), C-MMSE (b = .037, p < .001), episodic memory (b = .026, p < .001), Symbol Digit Modalities Test perceptual speed (b = .036, p < .001), and Digit Span Backward working memory (b = .047, p < .001) at Wave 2. Activity engagement, cognitive activity in particular, significantly mediates the effect of education on all cognitive tests, with the size of the mediating effect ranging from 16% to approximately 24%. Amount of schooling early in life is significantly related to late-life cognitive function in virtually all domains, and cognitive activity is one of many links between the two. J Am Geriatr Soc 67:S525-S531, 2019.


Development and validation of an HPLC-UV method for pazopanib quantification in human plasma and application to patients with cancer in routine clinical practice.
Pazopanib, a new oral angiogenesis inhibitor, has demonstrated clinical activity against multiple solid tumors and was approved for the treatment of patients with advanced renal cell carcinoma. As an exposure-response relationship has been observed for pazopanib, its therapeutic drug monitoring could be a valuable tool in clinical practice. Therefore, the aim of this study was to develop and validate a selective and precise high performance liquid chromatography-ultraviolet method for the measurement of pazopanib in plasma from patients with cancer. After liquid-liquid extraction with diethyl ether, pazopanib and gefitinib (internal standard) were separated using isocratic elution on an Ultrabase C18 column using a mobile phase consisting of a mixture in vol/vol proportion of 47:53 of ammonium acetate (pH, 7; 0.02 mol/L) and acetonitrile/methanol (70:30, vol/vol) pumped at a constant flow rate of 1 mL/min. Quantification was performed at 260 nm. Method validation was undertaken as per the guidelines for Bioanalytical Method Validation published by the Food and Drug Administration and European Medicines Agency. Calibration curves were linear over the range 0.5-100 mcg/mL. Interday and intraday coefficients of variations were less than 4.5%. The limit of detection and the lower limit of quantification were 0.2 and 0.5 mcg/mL, respectively. Recovery of pazopanib from plasma was >80%. This is the first high performance liquid chromatography-ultraviolet method for pazopanib quantification that has been validated within a wide range of plasma concentrations and is a suitable method for therapeutic drug monitoring of pazopanib.


A case of juvenile huntington disease in a 6-year-old boy.
Huntington disease is a neurodegenerative disorder distinguished by the triad of dominant inheritance, choreoathetosis and dementia, usually with onset in the fourth and fifth decades. It is caused by an unstable cytosine-adenine-guanine (CAG) trinucleotide repeat expansion in the gene IT15 in locus 4p16.3. Juvenile HD that constitutes about 3% to 10% of all patients is clinically different from adult-onset form and characterized by a larger number of CAG repeats typically exceeding 60. We report a case of a 6-year-old boy with myoclonic seizure and 140 CAG repeats confirmed by molecular genetic analysis.


Identification of a novel form of noradrenergic-dependent respiratory motor plasticity triggered by vagal feedback.
The respiratory control system is not just reflexive, it is smart, it learns, and, in fact, it has a memory. The respiratory system listens to and carefully remembers how previous stimuli affect breathing. Respiratory memory is laid down by adjusting synaptic strength between respiratory neurons. For example, repeated hypoxic bouts trigger a form of respiratory memory that functions to strengthen the ability of respiratory motoneurons to trigger contraction of breathing muscles. This type of respiratory plasticity is known as long-term facilitation (LTF). Although chemical feedback, such as hypoxia, initiates LTF, it is unknown whether natural modulation of mechanical feedback (from vagal inputs) also causes motor plasticity. Here, we used reverse microdialysis, electrophysiology, neuropharmacology, and histology to determine whether episodic modulation of vagally mediated mechanical feedback is able to induce respiratory LTF in anesthetized adult rats. We show that repeated obstructive apneas disrupt vagal feedback and trigger LTF of hypoglossal motoneuron activity and genioglossus muscle tone. This same stimulus does not cause LTF of diaphragm activity. Hypoxic episodes do not cause apnea-induced LTF; instead, LTF is triggered by modulation of vagal feedback. Unlike hypoxia-induced respiratory plasticity, vagus-induced LTF does not require 5-HT(2) receptors but instead relies on activation of α1-adrenergic receptors on hypoglossal motoneurons. In summary, we identify a novel form of hypoxia- and 5-HT-independent respiratory motor plasticity that is triggered by physiological modulation of vagal feedback and is mediated by α1-adrenergic receptor activation on (or near) hypoglossal motoneurons.


Developmental changes of the sensitivity of cardiac and liver mitochondrial permeability transition pore to calcium load and oxidative stress.
Opening of the mitochondrial membrane permeability transition pore (MPTP) is an important factor in the activation of apoptotic and necrotic processes in mammalian cells. In a previous paper we have shown that cardiac mitochondria from neonatal rats are more resistant to calcium load than mitochondria from adult animals. In this study we have analyzed the ontogenetic development of this parameter both in heart and in liver mitochondria. We found that the high resistance of heart mitochondria decreases from day 14 to adulthood. On the other hand, we did not observe a similar age-dependent sensitivity in liver mitochondria, particularly in the neonatal period. Some significant but relatively smaller increase could be observed only after day 30. When compared with liver mitochondria cardiac mitochondria were more resistant also to the peroxide activating effect on calcium-induced mitochondrial swelling. These data thus indicate that the MPTP of heart mitochondria is better protected against damaging effects of the calcium load and oxidative stress. We can only speculate that the lower sensitivity to calcium-induced swelling may be related to the higher ischemic tolerance of the neonatal heart.


A new approach to prostate cancer screening.
Prostate cancer screening based on prostate-specific antigen (PSA) testing has been a matter of controversy. Although screening for prostate cancer was effective in reducing mortality, it resulted in overdiagnosis, which translated into unnecessary treatments and numerous adverse effects. As a result, recommendations from scientific societies became increasingly restrictive. In the recent years, new approaches to prostate cancer screening have been proposed. These new approaches are aimed at solving the controversy between widespread screening vs. no screening, and reconsidering PSA testing as a screening tool with a good benefit/risk balance. In this context, the European Association of Urology submitted a proposal to the European Commission for prostate cancer screening to be performed as a function of baseline PSA concentrations. The European Commission recently recommended the implementation of organized prostate cancer screening programs for men aged ≤70 years based on PSA values in combination with follow-up magnetic resonance imaging.


Clinical Practice in the Prevention, Diagnosis and Treatment of Vitamin D Deficiency: A Central and Eastern European Expert Consensus Statement.
Vitamin D deficiency has a high worldwide prevalence, but actions to improve this public health problem are challenged by the heterogeneity of nutritional and clinical vitamin D guidelines, with respect to the diagnosis and treatment of vitamin D deficiency. We aimed to address this issue by providing respective recommendations for adults, developed by a European expert panel, using the Delphi method to reach consensus. Increasing the awareness of vitamin D deficiency and efforts to harmonize vitamin D guidelines should be pursued. We argue against a general screening for vitamin D deficiency but suggest 25-hydroxyvitamin D (25(OH)D) testing in certain risk groups. We recommend a vitamin D supplementation dose of 800 to 2000 international units (IU) per day for adults who want to ensure a sufficient vitamin D status. These doses are also recommended for the treatment of vitamin D deficiency, but higher vitamin D doses (e.g., 6000 IU per day) may be used for the first 4 to 12 weeks of treatment if a rapid correction of vitamin D deficiency is clinically indicated before continuing, with a maintenance dose of 800 to 2000 IU per day. Treatment success may be evaluated after at least 6 to 12 weeks in certain risk groups (e.g., patients with malabsorption syndromes) by measurement of serum 25(OH)D, with the aim to target concentrations of 30 to 50 ng/mL (75 to 125 nmol/L).


Unicystic ameloblastoma: a clinicopathologic study of 33 Chinese patients.
The term unicystic ameloblastoma refers to those cystic lesions that show clinical, radiographic, or gross features of a jaw cyst, but on histologic examination show a typical ameloblastomatous epithelium lining part of the cyst cavity, with or without luminal and/or mural tumor growth. To ascertain the clinicomorphologic spectrum and biologic behavior of this tumor group, the clinicopathologic features of 33 unicystic ameloblastomas from Chinese patients were studied. This series represents approximately 19% of all cases of ameloblastoma accessioned in the authors' hospital during a 15-year period. Twenty-one patients were male and 12 were female, for a total of 33 patients. The age at diagnosis ranged from 8 to 60 years (mean, 25.3 yrs) and peaked at the second and third decades (70%), Thirty tumors (91%) occurred in the mandible and three in the maxilla. Of the 29 patients with a radiographic record, an expansive unilocular radiolucency was seen in 22 cases, and was multilocular in seven cases. Microscopically, all tumors demonstrated a generally monocystic growth pattern. Eight tumors were simple cystic, 10 comprised intraluminal tumor nodules, and the remaining 15 had a conspicuous component of infiltrative tumor islands in the cyst capsule. The cystic tumor linings invariably showed, at least in part, a typical ameloblastomatous pattern that was often accompanied by epithelial areas of various histologic appearance. Follow up of 29 patients revealed no recurrence in less than 4 years of follow up, but did reveal a 35% recurrence rate at more than 4 years of follow up. The average interval to recurrence was approximately 7 years. Recurrence also appeared to relate to histologic subtypes of unicystic ameloblastoma, with those invading the fibrous wall having a rate of 35.7%, but other types having a rate of 6.7%. Despite the fact that unicystic ameloblastoma may, in general, compare favorably with its solid or multicystic counterpart in terms of clinical behavior and response to treatment, the subsets of the maxillary lesions or tumors containing invading islands in the fibrous wall could have a high risk of recurrence. Furthermore, recurrence of unicystic ameloblastoma may be long delayed, and a long-term postoperative follow up is essential to the proper management of these patients.


Frontonasal dysmorphology in bipolar disorder by 3D laser surface imaging and geometric morphometrics: comparisons with schizophrenia.
Any developmental relationship between bipolar disorder and schizophrenia engenders continuing debate. As the brain and face emerge in embryological intimacy, brain dysmorphogenesis is accompanied by facial dysmorphogenesis. 3D laser surface imaging was used to capture the facial surface of 13 male and 14 female patients with bipolar disorder in comparison with 61 male and 75 female control subjects and with 37 male and 32 female patients with schizophrenia. Surface images were analysed using geometric morphometrics and 3D visualisations to identify domains of facial shape that distinguish bipolar patients from controls and bipolar patients from those with schizophrenia. Both male and female bipolar patients evidenced significant facial dysmorphology: common to male and female patients was overall facial widening, increased width of nose, narrowing of mouth and upward displacement of the chin; dysmorphology differed between male and female patients for nose length, lip thickness and tragion height. There were few morphological differences in comparison with schizophrenia patients. That dysmorphology of the frontonasal prominences and related facial regions in bipolar disorder is more similar to than different from that found in schizophrenia indicates some common dysmorphogenesis. Bipolar disorder and schizophrenia might reflect similar insult(s) acting over slightly differing time-frames or slightly differing insult(s) acting over a similar time-frame.


Evolution of the vertebrate claudin gene family: insights from a basal vertebrate, the sea lamprey.
Claudins are major constituents of tight junctions, contributing both to their intercellular sealing and selective permeability properties. While claudins and claudin-like molecules are present in some invertebrates, the association of claudins with tight junctions has been conclusively documented only in vertebrates. Here we report the sequencing, phylogenetic analysis and comprehensive spatiotemporal expression analysis of the entire claudin gene family in the basal extant vertebrate, the sea lamprey. Our results demonstrate that clear orthologues to about half of all mammalian claudins are present in the lamprey, suggesting that at least one round of whole genome duplication contributed to the diversification of this gene family. Expression analysis revealed that claudins are expressed in discrete and specific domains, many of which represent vertebrate-specific innovations, such as in cranial ectodermal placodes and the neural crest; whereas others represent structures characteristic of chordates, e.g. pronephros, notochord, somites, endostyle and pharyngeal arches. By comparing the embryonic expression of claudins in the lamprey to that of other vertebrates, we found that ancestral expression patterns were often preserved in higher vertebrates. Morpholino mediated loss of Cldn3b demonstrated a functional role for this protein in placode and pharyngeal arch morphogenesis. Taken together, our data provide novel insights into the origins and evolution of the claudin gene family and the significance of claudin proteins in the evolution of vertebrates.


[Endoscopic radio wave hemostasis in combined treatment of gastroduodenal ulcers complicated by bleeding].
Radiowave hemostasis was used for endoscopic stopping of bleeding from gastric and duodenal ulcers in 96 patients. Primary single radiowave hemostasis permitted to achieve stable effect in 80 (83.3%) patients. After repeated radiowave hemostasis bleeding was completely stopped in 14 (14.6%) patients, 2 (2.1%) patients of this group underwent urgent surgery. On day 10 after radiowave hemostasis 95% of mucosal defects epithialized.


Natural host resistance and in vivo selection of malignant tumour cells.
In cells transformed in vitro by different agents we have identified two discrete characteristics essential for in vivo selection by the effectors of host natural resistance. These two biochemically distinct characteristics, ie resistance to hydrogen peroxide and ability to secrete E type prostaglandin (PGE) in contact with NK cells, macrophages and neutrophils, may both be expressed either in about 100% of cells transformed by Rous sarcoma virus, or in less than 0.1% of cells transformed spontaneously. The expression of these characteristics provides the cells with selective advantages in vivo and determines the level of their tumorigenic activity and experimental metastasis-forming activity. These characteristics are apparently necessary, but insufficient, for spontaneous metastasizing activity. Demonstrated NK resistance of malignant cell variants is directly connected with their ability to produce PGE. The in vitro transformed cells which do not possess these qualities apparently have no ability for in vivo survival and growth. Though these two cellular properties are connected with biochemically different pathways (catabolism of H2O2 and metabolism of arachidonic acid) they are jointly expressed, or not expressed, in transformed cells. The possible mechanisms of in vivo selection of malignant cell variants, with one or a few cell properties united in clusters, and the dual role of natural resistance in this process are discussed.


Nitric Oxide-Mediated Induction of Dispersal in Pseudomonas aeruginosa Biofilms Is Inhibited by Flavohemoglobin Production and Is Enhanced by Imidazole.
The biological signal molecule nitric oxide (NO) was found to induce biofilm dispersal across a range of bacterial species, which led to its consideration for therapeutic strategies to treat biofilms and biofilm-related infections. However, biofilms are often not completely dispersed after exposure to NO. To better understand this phenomenon, we investigated the response of Pseudomonas aeruginosa biofilm cells to successive NO treatments. When biofilms were first pretreated with a low, noneffective dose of NO, a second dose of the signal molecule at a concentration usually capable of inducing dispersal did not have any effect. Amperometric analysis revealed that pretreated P. aeruginosa cells had enhanced NO-scavenging activity, and this effect was associated with the production of the flavohemoglobin Fhp. Further, quantitative real-time reverse transcription-PCR (qRT-PCR) analysis showed that fhp expression increased by over 100-fold in NO-pretreated biofilms compared to untreated biofilms. Biofilms of mutant strains harboring mutations in fhp or fhpR, encoding a NO-responsive regulator of fhp, were not affected in their dispersal response after the initial pretreatment with NO. Overall, these results suggest that FhpR can sense NO to trigger production of the flavohemoglobin Fhp and inhibit subsequent dispersal responses to NO. Finally, the addition of imidazole, which can inhibit the NO dioxygenase activity of flavohemoglobin, attenuated the prevention of dispersal after NO pretreatment and improved the dispersal response in older, starved biofilms. This study clarifies the underlying mechanisms of impaired dispersal induced by repeated NO treatments and offers a new perspective for improving the use of NO in biofilm control strategies.


Enhancing vocational outcomes of spinal cord-injured persons: the occupational therapist's role.
Because work is a core element of our physical, social, and psychological survival, the significance of work for spinal cord-injured persons is no less than it is for able-bodied persons. To develop expectations of a productive life-style, vocational planning must be initiated early in the rehabilitation process, with the occupational therapist contributing significantly to the initial and ongoing functional and prevocational assessment. Interaction between the therapist and the spinal cord-injured person can promote the experience of control over environment, a feeling of responsibility for success of the rehabilitation process, the ability to solve functional problems outside of the rehabilitation environment, an understanding of the range of behavioral and environmental options available to the individual, and successful performance of job-related tasks in a supportive setting. The interaction between the occupational therapist, the vocational rehabilitation specialist, and the employer should be characterized by clear, nontechnical communication, an understanding of what functional activities the patient actually does (rather than what the person can do), a willingness to try creative solutions to environmental and performance problems, and a recognition of the employer's need for quantity and quality of production. All these combined reduces the potential for failure on the job and enhances the likelihood of achieving the highest possible level of vocational potential.


New media art design based on fast visual segmentation and 3D image processing.
Acquiring innovative styles and compositions from intricate and heterogeneous artistic imagery has emerged as a pivotal research quandry within contemporary new media art image conception. In a concerted effort to adeptly distill the quintessence of artistic styles and elements embedded within these visuals, an innovative methodology is posited herein, underpinned by an enhanced U-net segmentation framework and harmoniously fused with the surface extraction image reconstruction algorithm. This meticulous amalgamation endeavors to attain accurate segmentation and tridimensional reconstruction of the artistry encapsulated in these images. Primarily, the imagery is meticulously partitioned, culminating in an output that artfully encapsulates the inherent artistic attributes. Subsequently, this segmentation outcome is adeptly reconstituted, bestowing form to a three-dimensional artistry model. Empirical validation substantiates the efficacy of this approach, with the method's Mean Intersection over the Union (MIoU) parameter yielding an impressive score of 0.939 in segmentation performance. Moreover, the peak signal-to-noise ratio and structural similarity attain commendable zeniths of 38.16 and 0.9808, respectively, underscoring the excellence of the reconstruction process. The proposed methodology demonstrates its prowess in exacting segmentation and comprehensive reconstruction of semantic intricacies and nuanced features pervading the realm of artistic imagery. Consequently, this novel methodology augments artists' capacity to discern diverse artistic paradigms and fabricate superlative new media art compositions of heightened caliber.


Phase-contrast and magnification radiography at diagnostic X-ray energies using a pseudo-microfocus X-ray source.
To investigate the use of conventional diagnostic X-ray tubes for applications in which specialist microfocus sources are normally required. A conventional diagnostic X-ray tube was used in conjunction with a range of apertures to investigate improvements in spatial resolution using a line-pairs test object. Phase-contrast effects were investigated by varying source-to-object and object-to-receptor distances using a 2-French catheter as a clinically realistic test object. For magnification radiography using a computed radiography receptor and conventional X-ray tube with a 1-mm nominal focus size, the limiting spatial resolution was improved from 3.55 line-pairs per millimetre, for a conventional contact image, to 5.6 line-pairs per millimetre, for a ×2 magnified view with a 250-µm aperture. For inline phase-contrast radiography, phase contrast enhancement of a 2-French catheter was demonstrated, and the expected trends with variations in source-to-object and object-to-receptor distances were found. Images of a neonatal phantom demonstrated a subtle improvement in visibility of a superimposed 1-French catheter simulating a percutaneously inserted central catheter for no increase in patient radiation dose. Spatial resolution improvement and visible phase contrast can be produced in clinically relevant objects using a pseudo-microfocus geometry at X-ray energies in the normal diagnostic range, using conventional diagnostic X-ray tubes and image receptors. The disadvantages of the proposal are the large distances required to produce phase contrast and limitations imposed by the resulting tube loading. It is possible to use conventional diagnostic X-ray equipment in applications that normally require microfocus X-ray sources. This presents some possibilities for clinical applications.


STENTYS coronary system: current status and future direction.
Stentys self-apposing stent was designed to face complex lesions in the precincts of percutaneous coronary interventions. Nitinol platform and disconnectable struts were designed to provide a complete apposition on the vessel wall in challenging lesions such as significant tapering, primary angioplasty in ST segment elevation myocardial infarction and bifurcation. Stentys X-position S is a sirolimus eluting stent with a novel delivery system aiming to improve positioning. Clinical trials showed good results in terms of procedural success rate, clinical outcome and short-term strut apposition. Nevertheless, Stentys stent did not show superiority over the conventional balloon-expandable stents in the clinical outcomes. Authors underlined the importance of a learning curve and an adequate training period to get familiar with the device's features. Future trials in an all-comer population using the novel X-Position S stent will confirm the preliminary findings and strengthen evidence in clinical practice.


Socioeconomic Status and Incidence of Traffic Accidents in Metropolitan Tehran: A Population-based Study.
Population-based estimates of traffic accidents (TAs) are not readily available for developing countries. This study examined the contribution of socioeconomic status (SES) to the risk of TA among Iranian adults. A total of 64,200people aged ≥18years were identified from 2008 Urban Health Equity Assessment and Response Tool (Urban HEART) survey. 22,128 households were interviewed to estimate the overall annual incidence, severity and socioeconomic determinants of TAs for males and females in Iranian capital over the preceding year. Wealth index and house value index were constructed for economic measurement. Weighted estimates were computed adjusting for complex survey design. Logistic regression models were used to examine individual and SES measures as potential determinants of TAs in adults. The overall incidence of traffic accident was 17.3(95% CI 16.0, 18.7) per 1000 per year. TA rate in men and women was 22.6(95% CI 20.6, 24.8) and 11.8(95% CI 10.4, 13.2), respectively. The overall TA mortality rate was 26.6(95% CI 13.4, 39.8) per 100,000 person-years, which was almost three times higher in men than that for women (40.4 vs. 12.1 per 100,000person-years). Lower economic level was associated with increased incidence and mortality of TA. Association between SES and incidence, and severity and mortality of TA were identified. TAs occur more in lower socioeconomic layers of the society. This should be taken seriously into consideration by policy makers, so that preventive programs aimed at behavioral modifications in the society are promoted to decrease the health and economic burden imposed by TAs.


An algorithm for network-based gene prioritization that encodes knowledge both in nodes and in links.
Candidate gene prioritization aims to identify promising new genes associated with a disease or a biological process from a larger set of candidate genes. In recent years, network-based methods - which utilize a knowledge network derived from biological knowledge - have been utilized for gene prioritization. Biological knowledge can be encoded either through the network's links or nodes. Current network-based methods can only encode knowledge through links. This paper describes a new network-based method that can encode knowledge in links as well as in nodes. We developed a new network inference algorithm called the Knowledge Network Gene Prioritization (KNGP) algorithm which can incorporate both link and node knowledge. The performance of the KNGP algorithm was evaluated on both synthetic networks and on networks incorporating biological knowledge. The results showed that the combination of link knowledge and node knowledge provided a significant benefit across 19 experimental diseases over using link knowledge alone or node knowledge alone. The KNGP algorithm provides an advance over current network-based algorithms, because the algorithm can encode both link and node knowledge. We hope the algorithm will aid researchers with gene prioritization.


An improved method for liver diseases detection by ultrasound image analysis.
Ultrasound imaging is a popular and noninvasive tool frequently used in the diagnoses of liver diseases. A system to characterize normal, fatty and heterogeneous liver, using textural analysis of liver Ultrasound images, is proposed in this paper. The proposed approach is able to select the optimum regions of interest of the liver images. These optimum regions of interests are analyzed by two level wavelet packet transform to extract some statistical features, namely, median, standard deviation, and interquartile range. Discrimination between heterogeneous, fatty and normal livers is performed in a hierarchical approach in the classification stage. This stage, first, classifies focal and diffused livers and then distinguishes between fatty and normal ones. Support vector machine and k-nearest neighbor classifiers have been used to classify the images into three groups, and their performance is compared. The Support vector machine classifier outperformed the compared classifier, attaining an overall accuracy of 97.9%, with a sensitivity of 100%, 100% and 95.1% for the heterogeneous, fatty and normal class, respectively. The Acc obtained by the proposed computer-aided diagnostic system is quite promising and suggests that the proposed system can be used in a clinical environment to support radiologists and experts in liver diseases interpretation.


Percutaneously Inserted Central Catheter-Related Pleural Effusion in a Level III Neonatal Intensive Care Unit: A 5-Year Review (2008-2012).
Although peripherally inserted central catheters (PICCs) provide vascular access in newborns who require parenteral nutrition and medications, they can be associated with complications that lead to significant morbidity and mortality. To describe the characteristics of pleural effusion (PLE) associated with PICC use in a large level III neonatal intensive care unit. A retrospective review of PICC-related PLE in newborns was conducted over a 5-year period, from 2008-2012. A total of 926 PICCs were inserted, accounting for 17,606 catheter days. PICC-related PLE was identified in 7 infants, with an incidence of 0.4 per 1000 catheter days. Infants who developed PLE had a median gestational age of 28 weeks (range, 24-38 weeks) and birth weight of 735 g (range, 500-2975 g). PICCs were inserted at a median age of 4 days (range, 3-11 days). The median time from catheter insertion to the development of PLE was 16 days (range, 7-75 days). In all cases, the catheter tips were centrally located at the time of insertion but migrated to the subclavian veins or tributaries at the time of the events. PICC-related PLE can be associated with the migration of PICC tips to noncentral locations, despite optimal positioning of the tip at the time of insertion. Attention should be paid to migration of catheter tips on subsequent x-ray films. For PICCs inserted via upper limb or scalp, serial follow-up x-rays, beginning 1 week after insertion, may be helpful to detect migration of catheter tips and identify patients at risk.


Home Health Care For Children With Medical Complexity: Workforce Gaps, Policy, And Future Directions.
With the medical and surgical advances of recent decades, a growing proportion of children rely on home-based care for daily health monitoring and care tasks. However, a dearth of available home health care providers with pediatric training to serve children and youth with medical complexity markedly limits the current capacity of home health care to meet the needs of patients and their families. In this article we analyze the workforce gaps, payment models, and policy challenges unique to home health care for children and youth with medical complexity, including legal challenges brought by families because of home nursing shortages. We propose a portfolio of solutions to address the current failures, including payment reform, improved coordination of services and pediatric home health training through partnerships with child-focused health systems, telehealth-enabled opportunities to bridge current workforce gaps, and the better alignment of pediatric care with the needs of adult-focused long-term services and supports.


In situ expression of nifD in Geobacteraceae in subsurface sediments.
In order to determine whether the metabolic state of Geobacteraceae involved in bioremediation of subsurface sediments might be inferred from levels of mRNA for key genes, in situ expression of nifD, a highly conserved gene involved in nitrogen fixation, was investigated. When Geobacter sulfurreducens was grown without a source of fixed nitrogen in chemostats with acetate provided as the limiting electron donor and Fe(III) as the electron acceptor, levels of nifD transcripts were 4 to 5 orders of magnitude higher than in chemostat cultures provided with ammonium. In contrast, the number of transcripts of recA and the 16S rRNA gene were slightly lower in the absence of ammonium. The addition of acetate to organic- and nitrogen-poor subsurface sediments stimulated the growth of Geobacteraceae and Fe(III) reduction, as well as the expression of nifD in Geobacteraceae. Levels of nifD transcripts in Geobacteraceae decreased more than 100-fold within 2 days after the addition of 100 microM ammonium, while levels of recA and total bacterial 16S rRNA in Geobacteraceae remained relatively constant. Ammonium amendments had no effect on rates of Fe(III) reduction in acetate-amended sediments or toluene degradation in petroleum-contaminated sediments, suggesting that other factors, such as the rate that Geobacteraceae could access Fe(III) oxides, limited Fe(III) reduction. These results demonstrate that it is possible to monitor one aspect of the in situ metabolic state of Geobacteraceae species in subsurface sediments via analysis of mRNA levels, which is the first step toward a more global analysis of in situ gene expression related to nutrient status and stress response during bioremediation by Geobacteraceae.


Effect of mouse cumulus cells on the in vitro maturation and developmental potential of bovine denuded germinal vesicle oocytes.
We investigated the effect mouse cumulus cells (mCCs) on the in vitro maturation (IVM) and developmental potential of bovine denuded germinal vesicle oocytes (DOs). Cumulus-oocyte complexes (COCs), DOs and DOs cocultured with either mCCs (DOs + mCCs) or bovine cumulus cells (bCCs; DOs + bCCs) were subjected to IVM. The meiosis II (MII) rates of DOs, glutathione (GSH) contents, zona pellucida (ZP) hardening and parthenogenetic blastocyst rates of MII oocytes were determined. The relative expression levels of bone morphogenetic protein 15 (BMP-15) and growth differentiation factor 9 (GDF-9) in MII oocytes were measured using quantitative real-time polymerase chain reaction (PCR). mCCs significantly increased the MII rate of DOs from 53.5 ± 3.58% to 69.67 ± 4.72% (p < 0.05) but had no effect on the GSH content (2.17 ± 0.31 pmol/oocyte with mCCs, 2.14 ± 0.53 pmol/oocyte without mCCs). For the DOs + mCCs group, the BMP-15 and GDF-9 expression levels were significantly higher and the ZP dissolution time was significantly lower (162.49 ± 12.51 s) than that of the DOs group (213.95 ± 18.87 s; p < 0.05). The blastocyst rate of the DOs + mCCs group (32.56 ± 4.94%) was similar to that of the DOs group (31.75 ± 3.65%) but was significantly lower than that of the COCs group (43.52 ± 5.37%; p < 0.05). In conclusion, mCCs increased the MII rate of DOs and expression of certain genes in MII oocytes, and decreased the ZP hardening of MII oocytes, but could not improve their GSH content or developmental potential.


Anthocyanins of Hierbamora (Solanum nigrescens): Revealing their Nutraceutical Potential for Controlling Hypertriglyceridemia and Helicobacter pylori Viability.
We present the inhibitory properties of the Solanum nigrescens anthocyanin fraction (SNAF) and its major constituents on alpha-glucosidase (AG), pancreatic lipase (PL), HMG-CoA reductase, and ornithine decarboxylase (ODC). The effect of SNAF was simultaneously evaluated in ICR male mice exposed to triglyceride charge test (TCT). HPLC-MS profiling revealed the presence cyanidin-3-O-rutinoside-5-glucoside (CRG), delphinidin-3-(p-coumaroyl)-rutinoside-5-glucoside (DCRG), and petunidin-3-(cis-p-coumaroyl)-rutinoside-5-glucoside (PCRG) as major constituents of the fraction. SNAF, CRG, and specially PCRG, induced strong non-competitive inhibition on PL (IC50 , 33-86 μg mL-1 ). The results of TCT confirmed their capacity to ameliorate (p <0.001) hypertriglyceridemia during postprandial and interdigestive stages. SNAF, CRG, DCRG, and PCRG caused negligible growth inhibition (MIC>600 μg mL-1 ) on beneficial bacteria whereas SNAF and DCRG exerted inhibitory activity on Helicobacter pylori ATCC 53504 (MIC,187-64 μg mL-1 ). Additional exploration revealed that SNAF and DCRG produced non-competitive activity on H. pylori urease, which facilitates bacterial growth under acidic conditions.


What it's like to grow older: the aging perceptions of people with an intellectual disability in Ireland.
The Intellectual Disability Supplement to The Irish Longitudinal Study on Ageing is a national longitudinal study on the aging of people with an intellectual disability (ID) using a randomly selected sample of people with ID over the age of 40. In total, 367 people with an ID completed the aging perception self-report only section. Over 57% of people described their health as very good to excellent with no significant difference in health perceptions found for gender, level of ID, or living circumstance. Exploring people's perceptions utilizing PASW Text Analytics for Surveys 4.1 perceptions often supported negative views of the consequences of aging. These findings suggest challenging negative aging concepts is essential to promote positivity with associated improved health and wellbeing.


Characterization in rats of the anxiolytic potential of ELB139 [1-(4-chlorophenyl)-4-piperidin-1-yl-1,5-dihydro-imidazol-2-on], a new agonist at the benzodiazepine binding site of the GABAA receptor.
Benzodiazepines are among the most effective drugs for the treatment of anxiety disorders. However, their use is limited by undesired side effects, including sedation, development of tolerance, and drug abuse. The aim of this study was to evaluate the pharmacological profile of ELB139 [1-(4-chlorophenyl)-4-piperidin-1-yl-1,5-dihydro-imidazol-2-on] in different models of anxiety and to correlate these effects with its activity in vitro. ELB139 binds with an IC(50) of 1390 nM to the flunitrazepam binding site in rat forebrain cortical membranes. In rat hippocampal neurons, ELB139 potentiated GABA-induced currents without reaching the maximum effect of diazepam, indicating a partial benzodiazepine agonism. The potentiation was antagonized by the benzodiazepine antagonist flumazenil. ELB139 (10 and 30 mg/kg p.o.) was active in three different animal models of anxiety, i.e., in the elevated plus-maze, the light and dark box, and the Vogel conflict test. The anxiolytic activity in the elevated plus-maze was almost completely reversed by flumazenil (5 mg/kg i.p.), indicating that interaction with the benzodiazepine binding site is central to the pharmacological activity. No hint of sedation was observed at the doses tested in the three anxiety models and the open field. Also, no development of tolerance was observed within 6 weeks b.i.d. treatment with ELB139 in the elevated plus-maze test. In summary, ELB139 elicits strong effects on anxiety-related behavior in rats mediated by its benzodiazepine-like activity without showing sedation or the development of tolerance, a major side effect of benzodiazepines. These characteristics make the compound a prime candidate for clinical development.


Antibody-mediated osseous regeneration: a novel strategy for bioengineering bone by immobilized anti-bone morphogenetic protein-2 antibodies.
Bone regeneration often requires harvesting of autologous bone with significant potential morbidity and cost. Recombinant human bone morphogenetic protein (rhBMP)-2 has been approved by the U.S. Food and Drug Administration for specific regenerative indications. However, administration of exogenous growth factors has many drawbacks. The objective of the present proof-of-concept study was to determine whether immobilized anti-BMP-2 antibodies (Abs) could capture endogenous BMP-2 in local sites to mediate osteogenesis, a strategy we refer to as antibody-mediated osseous regeneration (AMOR). We have generated a murine anti-BMP-2 monoclonal antibody library, which was tested along with commercially available Abs in vitro and in vivo for their ability to mediate AMOR. In vitro studies demonstrated that only some anti-BMP-2 Abs tested formed immune complexes with BMP-2, which can bind to BMP cellular receptor, whereas other BMP-2/anti-BMP-2 complexes failed to bind. To investigate whether anti-BMP-2 Abs were able to mediate AMOR in vivo, anti-BMP-2 Abs were immobilized on absorbable collagen sponge (ACS) and surgically placed in rat calvarial defects. Microcomputed tomography analysis of live animals at 2, 4, and 6 weeks demonstrated that some anti-BMP-2 Abs immobilized on ACS mediated significant bone regeneration, whereas other clones did not mediate any bone regeneration. In situ BMP-2 and osteocalcin expression was investigated by immunohistochemistry. Results demonstrated higher BMP-2 and osteocalcin expression in sites with increased bone regeneration. Results provide first evidence for the ability of anti-BMP2 Abs to form an immune complex with endogenous BMP-2 and mediate bone regeneration in vivo, suggesting a promising therapeutic method for tissue engineering.


The Role and Potential Therapeutic Implications of the Fibroblast Growth Factors in Energy Balance and Type 2 Diabetes.
Obesity and its associated metabolic diseases have reached epidemic proportions worldwide, reducing life expectancy and quality of life. Several drugs have been tested to treat these diseases but many of them have damaging side effects. Consequently, there is an urgent need to develop more effective therapies. Recently, endocrine fibroblast growth factors (FGFs) have become attractive targets in the treatment of metabolic diseases. This review summarizes their most important functions as well as FGF-based therapies for the treatment of obesity and type 2 diabetes (T2D). Recent studies demonstrate that circulating levels of FGF19 are reduced in obesity. In fact, exogenous FGF19 administration is associated with a reduction in food intake as well as with improvements in glycaemia. In contrast, FGF21 levels are elevated in subjects with abdominal obesity, insulin resistance and T2D, probably representing a compensatory response. Additionally, elevated levels of circulating FGF23 in individuals with obesity and T2D are reported in most clinical studies. Finally, increased FGF1 levels in obese patients associated with adipogenesis have been described. FGFs constitute important molecules in the treatment of metabolic diseases due to their beneficial effects on glucose and lipid metabolism. Among all members, FGF19 and FGF21 have demonstrated the ability to improve glucose, lipid and energy homeostasis, along with FGF1, which was recently discovered to have beneficial effects on metabolic homeostasis. Additionally, FGF23 may also play a role in insulin resistance or energy homeostasis beyond mineral metabolism control. These results highlight the relevant use of FGFs as potential biomarkers for the early diagnosis of metabolic diseases. In this regard, notable progress has been made in the development of FGF-based therapies and different approaches are being tested in different clinical trials. However, further studies are needed to determine their potential therapeutic use in the treatment of obesity and obesity-related comorbidities.


Osteonecrosis of the Shoulders in Pediatric Patients Treated for Leukemia or Lymphoma: Single-Institutional Experience.
Osteonecrosis of the hips and knees is an often debilitating adverse event in children treated with glucocorticoids for leukemia and lymphoma but the impact of shoulder involvement has been understudied. Thus, we investigated the severity and functional impairment of shoulder osteonecrosis in a well-characterized population of pediatric patients treated for acute lymphoblastic leukemia or lymphoma. We retrospectively reviewed orthopaedic clinic and physical therapy evaluations to determine range of motion (ROM), pain, and impact of magnetic resonance-defined osteonecrosis (ON) on activities of daily living. Adverse events were classified according to the National Cancer Institute's Common Terminology Criteria for Adverse Events version 4.03. We identified 35 patients (22 female), median age at cancer diagnosis 14.2 (range, 4.3 to 19) years; median age at ON diagnosis 16.7 (range, 5.5 to 28) years. Median time to last follow-up from diagnosis of primary malignancy was 6.4 (range, 0 to 12.7) years and from diagnosis of ON was 4.0 (range, 0 to 8.9) years. Twenty-two patients had magnetic resonance evidence of ON; 43 shoulders involved at least 30% of the articular surface of the capital humeral epiphyses.Common Terminology Criteria for Adverse Events mean scores for initial assessments of 55 shoulders (29 patients) showed moderate negative impact of ON on activity of daily living (1.94), decreased ROM limiting athletic activity (0.98), and mild to moderate levels of pain (1.38). Analysis of this group's most recent assessment showed improvement in pain and ON over time, with an average pain grade of 0.58 indicating no pain to mild pain, and 1.37 for ON grade, indicating asymptomatic to mildly symptomatic impact on activities of daily living. We also found minimal worsening average ROM grades (1.11). Shoulder ON is an underappreciated adverse late effect of therapy in children treated for leukemia/lymphoma which can limit quality of life and functionality. In most cases, pain and disability can be improved with treatment. Level IV-case series.


Education's impact on primary care dentists' confidence in treating patients with special health care needs.
Special care dentistry (SCD) is still developing in XX. This study aimed to clarify whether primary care dentists are treating patients with special health care needs (SHCN), to know if they have had previous education on SCD (on an undergraduate or postgraduate level), whether their training level impacts their confidence when treating patients with SHCN, and to assess their opinion on SCD as a relevant topic in undergraduate education. A survey was answered by 149 primary care dentists working for the National Health Service of the XX region in XX, including information on their daily clinical practice, undergraduate, and postgraduate training in SCD, and their opinions on them. Most interviewees would like to complement their training and believed that SCD should be formally incorporated into undergraduate programs. There was a significant association between confidence in treating patients with SHCN and the rating of their undergraduate training, and between confidence and the number of hours of continuous development courses. Most primary care dentists treat patients with SHCN regularly. Therefore, including training in the undergraduate curriculum and afterward becomes necessary to increase their confidence when facing this challenging group of patients.


p53-Induced DNA bending: the interplay between p53-DNA and p53-p53 interactions.
Specific p53 binding-induced DNA bending and its underlying responsible forces are crucial for the understanding of selective transcription activation. Diverse p53-response elements exist in the genome; however, it is not known what determines the DNA bending and to what extent. In order to gain knowledge of the forces that govern the DNA bending, molecular dynamics simulations were performed on a series of p53 core domain tetramer-DNA complexes in which each p53 core domain was bound to a DNA quarter site specifically. By varying the sequence of the central 4-base pairs of each half-site, different DNA bending extents were observed. The analysis showed that the dimer-dimer interactions in p53 were similar for the complexes; on the other hand, the specific interactions between the p53 and DNA, including the interactions of Arg280, Lys120, and Arg248 with the DNA, varied more significantly. In particular, the Arg280 interactions were better maintained in the complex with the CATG-containing DNA sequence and were mostly lost in the complex with the CTAG-containing DNA sequence. Structural analysis shows that the base pairings for the CATG sequence were stable throughout the simulation trajectory, whereas those for the CTAG sequence were partially dissociated in part of the trajectory, which affected the stability of the nearby Arg280-Gua base interactions. Thus, DNA bending depends on the balance between the p53 dimer-dimer interactions and p53-DNA interactions, which is in turn related to the DNA sequence and DNA flexibility.


The Canadian Registry on Nonvariceal Upper Gastrointestinal Bleeding and Endoscopy (RUGBE): Endoscopic hemostasis and proton pump inhibition are associated with improved outcomes in a real-life setting.
From the Canadian Registry of patients with Upper Gastrointestinal Bleeding and Endoscopy (RUGBE), we determined clinical outcomes and explored the roles of endoscopic and pharmacologic therapies in a contemporary real-life setting. Analysis of randomly selected patients endoscoped for nonvariceal upper gastrointestinal bleeding at 18 community and tertiary care institutions between 1999 and 2002. Covariates and outcomes were defined a priori and 30-day follow-up obtained. Logistic regression models identified predictors of outcomes. One thousand eight-hundred and sixty-nine patients were included (66 +/- 17 yr, 38% female, 2.5 +/- 1.6 comorbid conditions, hemoglobin, 96 +/- 27 g/L, 54% received a mean of 2.9 +/- 1.7 units of blood). Endoscopy was performed within 24 h in 76%, with ulcers (55%) most commonly noted. High-risk endoscopic stigmata and endoscopic therapy were reported in 37%. Rebleeding, surgery, and mortality rates were 14.1%, 6.5%, and 5.4%, respectively. Decreased rebleeding was significantly and independently associated with PPI use (85% of patients, mean daily dose 56 +/- 53 mg) in all patients regardless of endoscopic stigmata, (odds ratio (OR):0.53, 95% confidence interval, 95% CI:0.37-0.77) and endoscopic hemostasis in patients with high-risk stigmata (OR:0.39, 95% CI:0.25-0.61). PPI use (OR:0.18, 95% CI:0.04-0.80) and endoscopic therapy (OR:0.31, 95% CI:0.11-0.91) were also each independently associated with decreased mortality in patients with high-risk stigmata. These results appear to confirm the protective role of endoscopic therapy in patients with high-risk stigmata, and suggest that acute use of PPIs may be associated with a reduction of rebleeding in all patients, and lower mortality in patients with high-risk stigmata. Independent prospective validation of these observational findings is now required.


Scaling up the functional response for spatially heterogeneous systems.
Scale transition theory is a framework for predicting regional population dynamics from local process functions and estimates of spatial heterogeneity. Using this framework, we estimated regional scale functional responses for a benthic predator-prey system in the Baltic Sea. Functional responses were based on laboratory experiments or field observations of stomach contents, and prey densities measured at a local scale (0.1 m(2)) or a regional scale (300 km(2)). Laboratory data overestimated consumption at high prey densities, whereas predictions based on local scale data tallied closely with consumption observed at the regional scale. The predicted regional functional response was different for increasing and decreasing prey densities, reflecting that predator and prey densities, as well as the covariance between them, exhibit oscillatory dynamics. We conclude that it is important to validate laboratory data with small-scale field observations and that scale transition is a powerful tool for scaling-up process functions in heterogeneous systems.


Effects of terbuthylazine on myocardial oxidative stress and ferroptosis via Nrf2/HO-1 signaling pathway in broilers.
Terbuthylazine (TBA) is one of the most commonly used and effective herbicides. However, due to its affinity for soil organic matter and water solubility, TBA can lead to biological health concerns. This study exposed broilers to TBA (0 mg/kg bw, 0.4 mg/kg bw, 4 mg/kg bw) for 28 days. The results showed significant pathological damage in broiler myocardial tissue, such as widening of the interstitial space, rupture of muscle fibers, and deposition of myocardial collagen fibers. In addition, Under the 0.4 mg/kg bw TBA exposure, myocardial oxidative stress was observed in broilers, which was accompanied by the activation of Nrf2/HO-1 pathway and the increased protein and mRNA levels of NQO1, NOX2 and SOD2 antioxidant enzymes. However, Nrf2/HO-1 protein and mRNA levels were reversed at 4 mg/kg bw TBA exposure. Meanwhile, the Nrf2/HO-1 mediated antioxidant defense was impaired. In contrast with the low dose, the protein and gene expression levels of NQO1, NOX2, and SOD2 were reduced in 4 mg/kg bw TBA group. The expression of GPX4 and SLC7A11 was significantly downregulated at both protein and mRNA levels. Beyond that, ACSL4 expression was significantly up-regulated, and the protein result was consistent with the mRNA expression, demonstrating the occurrence of ferroptosis. In general, TBA exposure activated the Nrf2/HO-1 pathway, resulting in ferroptosis. This study links ferroptosis to the Nrf2/HO-1 pathway, providing new insights into the potential role of TBA in myocardial toxicity.


Reflections on the 20th anniversary of Taiwan's single-payer National Health Insurance System.
On its twentieth anniversary, Taiwan's National Health Insurance (NHI) stands out as a high-performing single-payer national health insurance system that provides universal health coverage to Taiwan's 23.4 million residents based on egalitarian ethical principles. The system has encountered myriad challenges over the years, including serious financial deficits. Taiwan's government managed those crises through successive policy adjustments and reforms. Taiwan's NHI continues to enjoy high public satisfaction and delivers affordable modern health care to all Taiwanese without the waiting times in single-payer systems such as those in England and Canada. It faces challenges, including balancing the system's budget, improving the quality of health care, and achieving greater cost-effectiveness. However, Taiwan's experience with the NHI shows that a single-payer approach can work and control health care costs effectively. There are lessons for the United States in how to expand coverage rapidly, manage incremental adjustments to the health system, and achieve freedom of choice.


Mutations affecting the synthesis of NADP-dependent glutamate dehydrogenase in Pseudomonas aeruginosa.
NADP-dependent glutamate dehydrogenase (NADP-GDH) was purified to homogeneity from Pseudomonas aeruginosa strain 8602 (PAC 1). The Mr determined by Sephadex gel filtration was 280,000; the subunit Mr determined by SDS-PAGE was 45,000. Mutant strains lacking NADP-GDH and glutamate synthase (Gdh-Glt-) required glutamate for growth. Transductants that lacked only NADP-GDH were indistinguishable from the wild-type strain in growth properties. It was concluded that NADP-GDH is not essential for growth of the wild-type organism and that glutamate formation via NAD-dependent glutamate dehydrogenase does not occur to a significant extent. A mutant strain, 39, producing high NADP-GDH activity, synthesized normal NADP-GDH and had the same intracellular glutamate concentrations as its parent. The mutation responsible for the synthesis of high levels of NADP-GDH was shown, by transduction, to be closely linked to the NADP-GDH structural gene (gdhA).


Antiallodynic effect of β-caryophyllene on paclitaxel-induced peripheral neuropathy in mice.
Painful peripheral neuropathy is a common side effect of paclitaxel (PTX). The use of analgesics is an important component for management of PTX-induced peripheral neuropathy (PINP). However, currently employed analgesics have several side effects and are poorly effective. β-caryophyllene (BCP), a dietary selective CB2 agonist, has shown analgesic effect in neuropathic pain models, but its role in chemotherapy-induced neuropathic pain has not yet been investigated. Herein, we used the mouse model of PINP to show the therapeutic effects of BCP in this neuropathy. Male Swiss mice receiving PTX (2 mg kg-1, ip, four alternate days) were treated with BCP (25 mg kg-1, po, twice a day) either during or after PTX administration. Some groups were also pretreated with AM630 (CB2 antagonist, 3 mg kg-1, ip) or AM251 (CB1 antagonist, 1 mg kg-1, ip). Spinal cord samples were collected in different time points to perform immunohistochemical analysis. BCP attenuated the established mechanical allodynia induced by PTX (p < 0.0001) in a CB2-dependent manner. Of note, when given concomitantly with PTX, BCP was able to attenuate the development of PINP (p < 0.0001). Spinal cord immunohistochemistry revealed that preventive treatment with BCP reduced p38 MAPK and NF-κB activation, as well as the increased Iba-1 and IL-1β immunoreactivity promoted by PTX. Our findings show that BCP effectively attenuated PINP, possibly through CB2-activation in the CNS and posterior inhibition of p38 MAPK/NF-κB activation and cytokine release. Taken together, our results suggest that BCP could be used to attenuate the establishment and/or treat PINP.


PGRN induces impaired insulin sensitivity and defective autophagy in hepatic insulin resistance.
Progranulin (PGRN) has recently emerged as an important regulator for glucose metabolism and insulin sensitivity. However, the underlying mechanisms of PGRN in the regulation of insulin sensitivity and autophagy remain elusive. In this study, we aimed to address the direct effects of PGRN in vivo and to evaluate the potential interaction of impaired insulin sensitivity and autophagic disorders in hepatic insulin resistance. We found that mice treated with PGRN for 21 days exhibited the impaired glucose tolerance and insulin tolerance and hepatic autophagy imbalance as well as defective insulin signaling. Furthermore, treatment of mice with TNF receptor (TNFR)-1 blocking peptide-Fc, a TNFR1 blocking peptide-Fc fusion protein to competitively block the interaction of PGRN and TNFR1, resulted in the restoration of systemic insulin sensitivity and the recovery of autophagy and insulin signaling in liver. Consistent with these findings in vivo, we also observed that PGRN treatment induced defective autophagy and impaired insulin signaling in hepatocytes, with such effects being drastically nullified by the addition of TNFR1 blocking peptide -Fc or TNFR1-small interference RNA via the TNFR1-nuclear factor-κB-dependent manner, indicating the causative role of PGRN in hepatic insulin resistance. In conclusion, our findings supported the notion that PGRN is a key regulator of hepatic insulin resistance and that PGRN may mediate its effects, at least in part, by inducing defective autophagy via TNFR1/nuclear factor-κB.


Tibial artery duplex ultrasound-derived peak systolic velocities may be an objective performance measure after above-knee endovascular therapy for arterial stenosis.
The ankle-brachial index (ABI) is a well-established measure of distal perfusion in lower extremity ischemia; however, the ABI is of limited value in patients with noncompressible lower extremity arteries. We sought to demonstrate whether duplex ultrasound-determined tibial artery velocities can be used as an alternative to ABI as an objective performance measure after endovascular treatment of above-knee arterial stenosis. Thirty-six patients undergoing above-knee endovascular intervention had preprocedure and postprocedure duplex ultrasound examination within 6 months of intervention. Preprocedure vs postprocedure changes in tibial artery mean peak systolic velocity (PSV; mean of proximal, mid, and distal velocities) were compared with changes in ABI and a reference (control) cohort of 68 patients without peripheral vascular disease. Thirty-six patients (41 limbs) had an above-knee endovascular intervention and had preprocedure and postprocedure duplex ultrasound examinations of the ipsilateral extremity including the tibial arteries. Before the procedure, mean tibial artery PSVs in the 36 patients undergoing intervention were outside (below) the 95% confidence intervals for the control patients. In comparing preprocedure and postprocedure PSVs, the mean anterior tibial (P < .01), mean peroneal (P < .01), and mean posterior tibial (P < .01) PSVs all increased and correlated with an increase in ABI (P < .01). After endovascular intervention, duplex ultrasound-derived mean PSVs fell within or near established reference ranges for patients without peripheral arterial disease. Mean tibial artery PSV increases were similar in patients with and without noncompressible vessels. Tibial artery PSVs increase, correlate with an increase in ABI, and fall within or near confidence intervals for normal controls after above-knee endovascular interventions. After endovascular intervention, tibial artery PSVs can supplement ABI as an objective performance measure in patients with and in particular without compressible tibial arteries.


Footwear in the causation and prevention of foot ulcers in diabetes mellitus.
Inappropriate footwear may be a major cause of foot ulceration among patients with diabetic neuropathy in India. No study has specifically examined the types of footwear or its components in patients with diabetes mellitus and their role in causing foot ulcers. We analysed the role of commonly used footwear in India in inducing first foot ulcers (FFU) in people with diabetes. Of 4800 patients with diabetes attending our centre over 1 year, 301 had FFU from different causes. Sixty-six patients with diabetic neuro-/vasculopathy presenting with ulcers due to footwear were included as cases. An equal number of patients with diabetes but without foot ulcers were enrolled as controls. Cases and controls were matched demographically and clinically for type of diabetes, metabolic control, duration of diabetes, comorbid conditions and foot neurovascular status. We did a detailed foot examination for neurological, vascular and wound status. We also evaluated the footwear in both groups. In one-fifth of 335 limbs (301 patients), the primary cause for the FFU was use of inappropriate footwear. The patients used seven different models of footwear, six of which were found to be inappropriate. The straps of footwear caused over 50% of ulcers. Another one-third were due to penetration of sharp objects through the outer sole of footwear; among these cases, 1 3.6% of ulcers were caused by not using soft inner soles. The use of softer insole is least effective in preventing foot ulcers. Similarly, straps contribute to a higher percentage of foot ulcers. Foot ulcers can be prevented by a combination of soft insole, with midsole and hard outsole with proper back counter and adjustable front and back straps.


Validation and Reliability of the 12-item Zarit Burden Interview among Informal Caregivers of Elderly Persons in Nigeria.
Research on ageing in Nigeria has shown that informal care places economic and psychological strain on the caregivers of the elderly persons. However, objective method of assessment of the burden of care for the elderly among caregivers is lacking. This study was aimed at validating the 12-item Zarit Burden Interview (ZBI) among caregivers of elderly persons in Nigeria in correlation with the Katz Index of Independence in Activities of Daily Living (Katz-ADL), the General Health Questionnaire (GHQ-12) and the Modified Conflicts Tactics Scale for Elder Abuse (MCTS). Eighty primary caregivers of community-dwelling elders were selected using a two-stage cluster sampling from two communities. Principal component analysis with varimax rotation was used to determine the factor structure of the ZBI. Construct validity was assessed using Spearman's correlations between the ZBI, the GHQ-12, Katz ADL and MCTS. Internal consistency and instrument reliability were examined with the Cronbach's alpha split-half correlation. Three factors with eigenvalues greater than 1.0 were extracted on the exploratory factor analysis comprising all 12 items accounting for 68.0% of the total item variance. The reliability analysis gave a Cronbach's α of 0.90 and a split-half correlation coefficient of 0.84. Spearman's correlation (rs) showed good correlation between the ZBI scores and GHQ-12 (rs=0.44, p<0.001), MCTS (rs=0.43, p<0.001) and Katz ADL (rs= -0.50, p <0.001). The 12-item ZBI is a valid and reliable tool for assessing burden in informal settings among caregivers of elderly persons. The tool can be used to assess burden of care among caregivers of elderly persons in community settings.