Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenes.
A major obstacle to creating precisely expressed transgenes lies in the epigenetic effects of the host chromatin that surrounds them. Here we present a strategy to overcome this problem, employing a Gal4-inducible luciferase assay to systematically quantify position effects of host chromatin and the ability of insulators to counteract these effects at phiC31 integration loci randomly distributed throughout the Drosophila genome. We identify loci that can be exploited to deliver precise doses of transgene expression to specific tissues. Moreover, we uncover a previously unrecognized property of the gypsy retrovirus insulator to boost gene expression to levels severalfold greater than at most or possibly all un-insulated loci, in every tissue tested. These findings provide the first opportunity to create a battery of transgenes that can be reliably expressed at high levels in virtually any tissue by integration at a single locus, and conversely, to engineer a controlled phenotypic allelic series by exploiting several loci. The generality of our approach makes it adaptable to other model systems to identify and modify loci for optimal transgene expression.


Is greater antibiotic therapy prior to ESS associated with differences in surgical outcomes in CRS?
Antibiotics have been a mainstay of chronic rhinosinusitis therapy; however, data suggest that antibiotics may also result in several adverse unintended consequences. We aimed to evaluate if greater antibiotic use prior to sinus surgery was associated with differences in surgical outcomes. Adult CRS patients enrolled in a prospective, multi-institutional, observational cohort study were asked to recall systemic antibiotic use in the 90 days prior to endoscopic sinus surgery (ESS). Antibiotic use was examined in relation to demographics, disease characteristics, and outcomes. Data were collected for 271 study participants followed for a mean of 14.9 [± 5.1] months across four institutions, with a mean preoperative antibiotic use of 27.8 [± 22.7] days out of the 90 preceding ESS. After ESS, significant improvement (P < 0.001) was reported for patient-reported outcome measures and endoscopy scores for the overall cohort. No bivariate correlation between preoperative antibiotic use and degree of benefit in objective clinical measures (endoscopy, Brief Smell Identification Test) was seen. Increased preoperative antibiotic use was associated with less improvement in 22-item SinoNasal Outcome Test (SNOT-22) and its rhinologic subdomain after ESS. Prevalence of achieving a minimal clinically important difference in overall SNOT-22 symptom scores was lower in those using more preoperative antibiotics, although statistical significance was not reached when adjusting for independent covariates in multivariate modeling. Higher amounts of previous antibiotic use do not appear to be associated with better ESS outcomes. Specific recommendations for antibiotic use as part of CRS-appropriate medical therapy prior to ESS require further study, particularly given concerns for antibiotic overuse and implications for improving outcomes in the modern healthcare era. 2C Laryngoscope, 129:558-566, 2019.


Number concepts of preschool children.
Using the format of the tin game described by Hughes in 1986, this study investigated the extent and variety of written representations of small numbers of objects used spontaneously and meaningfully by a sample of 39 preschool children.


[Hand grip strength in a sample of 11 to 14 years old children].
The determination of hand grip strength is an objective method of evaluation of upper members' integrity. It is also used as a general health indicator. Data about grip strength values in healthy children is useful in the research and diagnosis of diseases that interfere with grip force. The aim of this study was to determine average values of children hand grip strength by age and evaluate its relation with weight and height. Hand grip strength was measured with the dynamometer Jamar Hydraulic Hand in a convenience sample of 195 children of either gender, with ages between 11 and 14 years old. Statistical significant differences were observed in hand grip strength between genders, being higher in boys. Moderate correlations were found between grip strength and age (R = 0.679, p < 0.001), height (R = 0.691, p < 0.001), and weight (R = 0.620, p < 0.001). The present study presents values of hand grip strength obtained from a sample of Portuguese children of 11 to 14 years old, which are directly associated with height and weight.


[Need for research collaboration between cardiologists and emergency service doctors].
Time is a key factor when tackling acute coronary diseases and syndromes. The current organization of health services means that the first minutes of care are not carried out in cardiology rooms and do not depend on cardiologists. Instead they take place in the pre-hospital setting of emergency medical services (EMS) and in hospital accident and emergency services (A&E) and in the first phase this attention is provided by emergency doctors. Therefore, to obtain the best possible result for the patient it is necessary that there should be extensive two-way collaboration between cardiologists and emergency service doctors. This article analyzes these needs and tries to identify areas of improvement in two of the most frequent acute coronary processes: acute coronary syndrome and acute cardiac insufficiency.


Experimental estimation of the efficacy of the FLOTAC basic technique.
The FLOTAC technique is a quantitative coproscopic method for the diagnosis of parasitic infection that is based on the centrifugation of a fecal sample to levitate helminth eggs with a flotation solution in a proprietary apparatus. Determination of the efficacy of the FLOTAC method and multiplication factors for calculation of the number of Toxocara, Trichuris, and Ascaris eggs in 1 g of feces on the basis of the number of detected eggs is presented. An investigation was conducted using feces samples enriched with a known number of parasite eggs: 3, 15, 50, or 100 parasite eggs of 3 nematode genera (Toxocara, Trichuris, and Ascaris) per 1 g (EPG) of feces. In addition, 80 samples of dog feces were prepared consisting of 20 repetitions for each level of contamination. The samples were analyzed using the FLOTAC basic technique. The limit of detection was calculated as the lowest level of egg content at which at least 50% of repetitions were positive. Multiplication factors for estimating the true number of parasite eggs in the samples were derived from regression coefficients that illustrated the linear relationship between the number of detected eggs and the number of eggs added to the sample. The percentages of recovered eggs for 1 chamber and for the whole apparatus ranged from 11.67 to 21.90% and from 21.33 to 40.10%, respectively, depending on dose enrichment and genus of parasite. The limit of detection calculated for the whole FLOTAC device was 3 EPG and was 15 EPG for 1 chamber for each of the 3 parasite genera. The limit of quantification calculated for whole FLOTAC was 15 EPG for each of 3 kinds of eggs. For 1 chamber, the limit of quantification was 15 EPG for Ascaris and Toxocara eggs and 50 EPG for Trichuris eggs. Multiplication factors for calculation of the number of eggs in 1 g of feces calculated for whole FLOTAC were 3 (for Toxocara and Ascaris eggs) and 4 (for Trichuris eggs). Experimentally calculated parameters of the method differ significantly from the theoretical assumptions of the authors of the FLOTAC technique and can significantly affect the reliability of the results. This does not alter the fact that the FLOTAC technique is the most effective parasitological quantitative method, which can be used to detect parasitic forms in feces. However, the results of our study emphasized the need for validation of the method before using it in the laboratory.


Differential expression of proteins in fetal brains of alcohol-treated prenatally C57BL/6 mice: a proteomic investigation.
Alcohol is known to impede the growth of the central nervous system and to induce neurodegeneration through cellular apoptosis. We have previously shown that moderate prenatal alcohol exposure results in brain defects at different stages of development. In this study, we further characterize the proteomic architecture underlying ethanol teratogenesis during early fetal brain development using chromatography in conjunction with a LC-MS/MS system. Pregnant C57BL/6 mice were exposed from embryonic day 7 (E7) to E13 with either a 25% ethanol derived calorie or pair-fed liquid diets. At E13, fetal brains were collected from five dams for each group. Individual brains were homogenized and the extracted proteins were then tryptically digested and analyzed by LC-MS/MS. Label-free quantitative proteomic analyses were performed on proteomes extracted from fetal brains of both alcohol-treated (ALC) and pair-fed groups. These analyses demonstrated that prenatal alcohol exposure induced significant downregulation (p<0.001) of the expression of mitochondrial enzymes including ADP/ATP translocase 1, ATP synthase subunit alpha and ubiquinol-cytochrome-c reductases. In addition, mitochondrial carrier homolog 1, which plays a role in apoptosis, was significantly downregulated (p<0.001) in the ALC group. Moreover, among the cytosolic proteins that were significantly downregulated (p<0.001) are Bcl-2, 14-3-3 protein and calmodulin. Significant downregulation (p<0.001) of proteins that are critical for fetal brain development was observed such as prohibitin and neuronal migration protein doublecortin. These findings provide information about possible mechanisms underlying the effects of prenatal alcohol exposure during early embryonic stage.


Resistance to apoptosis in complicated Crohn's disease: Relevance in ileal fibrosis.
The stiffening of the extracellular matrix, and changes in its cellular and molecular composition, have been reported in the pathogenesis of fibrosis. We analyze the mechanisms that perpetuate ileal fibrosis in surgical resections of complicated Crohn's disease patients. Ileal resections were obtained from affected and non-affected tissue of stenotic or penetrating Crohn's disease behavior. Ilea from non-IBD patients were used as control tissue. All samples underwent RNA sequencing. Human small intestinal fibroblasts were treated for 48 h with IL-1β, TFGβ1, PDGFB or TNF-α. Resistance to apoptosis was analysed by RT-PCR, western blot and immunohistochemistry in ileal tissue and by RT-PCR and FACS in cultured cells. Growth factor-driven signaling pathways and increased RAS GTPase activity were up-regulated in affected ilea in which we found expression of both the antiapoptotic molecule MCL1 and the transcription factor ETS1 in submucosal fibroblasts, and a senescence-associated secretory phenotype. In cultured intestinal fibroblasts, PDGFB induced an ETS1-mediated resistance to apoptosis that was associated with the induction of both of TGFB1 and IL1B, a cytokine that replicated the expression of SASP detected in ileal tissue. ETS1 drove fibroblast polarization between inflammatory and fibrogenic phenotypes in IL1β-treated cells. Our data show resistance to apoptosis in complicated ileal CD, and demonstrate that PDGFB induce an ETS1-mediated resistance to apoptosis associated with an inflammatory and fibrogenic pattern of expression in intestinal fibroblasts. Results point to PDGFRB, IL1R1 or MCL1 as potential targets against ileal fibrosis.


Opioid requirement, opioid receptor expression, and clinical outcomes in patients with advanced prostate cancer.
Preclinical studies show that opioids stimulate angiogenesis and tumor progression through the mu opioid receptor (MOR). Although MOR is overexpressed in several human malignancies, the effect of chronic opioid requirement on cancer progression or survival has not been examined in humans. We performed a retrospective analysis on 113 patients identified in the Minneapolis VA Tumor Registry (test cohort) and 480 patients from the national VA Central Cancer Registry (validation cohort) who had been diagnosed with stage IV prostate cancer between 1995 and 2010 to examine whether MOR expression or opioid requirement is associated with disease progression and survival. All opioids were converted to oral morphine equivalents for comparison. Laser scanning confocal microscopy was used to analyze MOR immunoreactivity in prostate cancer biopsies. The effects of variables on outcomes were analyzed in univariable and multivariable models. In patients with metastatic prostate cancer, MOR expression and opioid requirement were independently associated with inferior progression-free survival (hazard ratio [HR] 1.65, 95% confidence interval [CI] 1.33-2.07, P<.001 and HR 1.08, 95% CI 1.03-1.13, P<.001, respectively) and overall survival (HR 1.55, 95% CI 1.20-1.99, P<.001 and HR 1.05, 95% CI 1.00-1.10, P = .031, respectively). The validation cohort confirmed that increasing opioid requirement was associated with worse overall survival (HR 1.005, 95% CI 1.002-1.008, P = .001). Higher MOR expression and greater opioid requirement are associated with shorter progression-free survival and overall survival in patients with metastatic prostate cancer. Nevertheless, clinical practice should not be changed until prospective randomized trials show that opioid use is associated with inferior clinical outcomes, and that abrogation of the peripheral activities of opioids ameliorates this effect.


Design and execution of clinical trials in orthopaedic surgery.
High-quality randomised controlled trials (RCTs) evaluating surgical therapies are fundamental to the delivery of evidence-based orthopaedics. Orthopaedic clinical trials have unique challenges; however, when these challenges are overcome, evidence from trials can be definitive in its impact on surgical practice. In this review, we highlight several issues that pose potential challenges to orthopaedic investigators aiming to perform surgical randomised controlled trials. We begin with a discussion on trial design issues, including the ethics of sham surgery, the importance of sample size, the need for patient-important outcomes, and overcoming expertise bias. We then explore features surrounding the execution of surgical randomised trials, including ethics review boards, the importance of organisational frameworks, and obtaining adequate funding. Cite this article: Bone Joint Res 2014;3:161-8.


Macrophage targeted theranostic strategy for accurate detection and rapid stabilization of the inflamed high-risk plaque.
Rationale: Inflammation plays a pivotal role in the pathogenesis of the acute coronary syndrome. Detecting plaques with high inflammatory activity and specifically treating those lesions can be crucial to prevent life-threatening cardiovascular events. Methods: Here, we developed a macrophage mannose receptor (MMR)-targeted theranostic nanodrug (mannose-polyethylene glycol-glycol chitosan-deoxycholic acid-cyanine 7-lobeglitazone; MMR-Lobe-Cy) designed to identify inflammatory activity as well as to deliver peroxisome proliferator-activated gamma (PPARγ) agonist, lobeglitazone, specifically to high-risk plaques based on the high mannose receptor specificity. The MMR-Lobe-Cy was intravenously injected into balloon-injured atheromatous rabbits and serial in vivo optical coherence tomography (OCT)-near-infrared fluorescence (NIRF) structural-molecular imaging was performed. Results: One week after MMR-Lobe-Cy administration, the inflammatory NIRF signals in the plaques notably decreased compared to the baseline whereas the signals in saline controls even increased over time. In accordance with in vivo imaging findings, ex vivo NIRF signals on fluorescence reflectance imaging (FRI) and plaque inflammation by immunostainings significantly decreased compared to oral lobeglitazone group or saline controls. The anti-inflammatory effect of MMR-Lobe-Cy was mediated by inhibition of TLR4/NF-κB pathway. Furthermore, acute resolution of inflammation altered the inflamed plaque into a stable phenotype with less macrophages and collagen-rich matrix. Conclusion: Macrophage targeted PPARγ activator labeled with NIRF rapidly stabilized the inflamed plaques in coronary sized artery, which could be quantitatively assessed using intravascular OCT-NIRF imaging. This novel theranostic approach provides a promising theranostic strategy for high-risk coronary plaques.


Aromatase inhibitors: a new paradigm in breast cancer treatment.
Microsomal cytochrome P450 (CYP 450) enzyme aromatase belongs to CYP 19 super family. It is involved in the conversion of androgens to estrogens. In postmenopausal women the main sites of aromatisation are skin, adipose tissue and breast. Aromatase localized in breast tumor produces sufficient estrogen for its proliferation. Hence it is an important target for the treatment of hormone dependent breast cancer in postmenopausal women. There are mainly two types of aromatase inhibitors, one is steroidal another is nonsteroidal type. The first and second generation aromatase inhibitors encounter the undesirable drug- drug interactions besides being not very specific and plagued with pharmacokinetic problems. Third generation aromatase inhibitors developed recently are more potent and specific with a greater capacity to annihilate circulating estrogen levels. These agents have satisfactory pharmacokinetic profiles and are devoid of major drug-drug interactions. Third generation aromatase inhibitors became drugs of choice for both first and second line treatment of advanced breast cancer. Aromatase inhibitors can also be used for neoadjuvant therapy of breast cancer in which they have achieved better therapeutic efficacy than tamoxifen. Early results of ATAC (Armidex Tamoxifen Alone or Combination) trial suggest that anastrozole is superior to tamoxifen in adjuvant setting for disease free survival, particularly in receptor positive patients, and in reducing the incidence of contralateral breast cancer. Therapeutic potential of aromatase inhibitors stretches beyond the postmenopausal breast cancer treatment as they also play a role in the treatment of estrogen dependent benign and malignant conditions such as gynaecomastia, prostate cancer, fibroadenomata and the induction of ovulation. By virtue of their ability to reduce estrogen levels they pose problems like demineralization of bone, hot flushes and anti-implantation effects.


The Chicago Health Corps: strengthening communities through structured volunteer service.
The Chicago Health Corps is an AmeriCorps*USA program, established in 1994 by the Corporation for National Service in partnership with the Health Resources and Services Administration (HRSA) of the U.S. Public Health Service. The Chicago Health Corps deploys 20 full-time equivalent corps members in selected community sites that offer primary health care services to Chicago's underserved families. Chicago Health Corps members provide a combination of outreach, home visit, and case management services to address unmet health needs identified by community members, including both laypersons and professionals. Providing meaningful opportunities for participants to assist their communities with health care helps corps members develop an awareness of their fellow community members and an ethic of service.


Phylogenomic Resolution of the Cetacean Tree of Life Using Target Sequence Capture.
The evolution of cetaceans, from their early transition to an aquatic lifestyle to their subsequent diversification, has been the subject of numerous studies. However, although the higher-level relationships among cetacean families have been largely settled, several aspects of the systematics within these groups remain unresolved. Problematic clades include the oceanic dolphins (37 spp.), which have experienced a recent rapid radiation, and the beaked whales (22 spp.), which have not been investigated in detail using nuclear loci. The combined application of high-throughput sequencing with techniques that target specific genomic sequences provide a powerful means of rapidly generating large volumes of orthologous sequence data for use in phylogenomic studies. To elucidate the phylogenetic relationships within the Cetacea, we combined sequence capture with Illumina sequencing to generate data for $\sim $3200 protein-coding genes for 68 cetacean species and their close relatives including the pygmy hippopotamus. By combining data from $>$38,000 exons with existing sequences from 11 cetaceans and seven outgroup taxa, we produced the first comprehensive comparative genomic data set for cetaceans, spanning 6,527,596 aligned base pairs (bp) and 89 taxa. Phylogenetic trees reconstructed with maximum likelihood and Bayesian inference of concatenated loci, as well as with coalescence analyses of individual gene trees, produced mostly concordant and well-supported trees. Our results completely resolve the relationships among beaked whales as well as the contentious relationships among oceanic dolphins, especially the problematic subfamily Delphinidae. We carried out Bayesian estimation of species divergence times using MCMCTree and compared our complete data set to a subset of clocklike genes. Analyses using the complete data set consistently showed less variance in divergence times than the reduced data set. In addition, integration of new fossils (e.g., Mystacodon selenensis) indicates that the diversification of Crown Cetacea began before the Late Eocene and the divergence of Crown Delphinidae as early as the Middle Miocene. [Cetaceans; phylogenomics; Delphinidae; Ziphiidae; dolphins; whales.].


Campylobacter jejuni and salmonella in raw red meats: A Public Health Laboratory Service Survey.
Thirty-one laboratories examined a total of 6169 meat samples, 1236 from abattoirs and 4933 from retail and other outlets. Campylobacter jejuni was isolated from 98 (1.6%). A higher isolation rate of 49/1236 (4.0%) was found among abattoir than among retail and other samples (49/4933-1.0%). Twenty-two of the laboratories looked for salmonella; although 94/4002 (2.3%) were positive, in only one sample of minced beef were campylobacter and salmonella found together. Isolation rates for salmonellae were 75/3576 (2.1%) from retail and 19/426 (4.5%) from abattoir samples.Analysis of the results revealed that (1) the contamination rate of raw red meat by C. jejuni is, in general, very low; (2) when contaminated, numbers of organisms are generally also very low; (3) enrichment procedures were of some value; 41/98 (42%) isolates were detected by enrichment only, but, on the other hand 8 (8%) were direct plate positive/enrichment negative; (4) practice at looking for the organism and increased seasonal temperatures over the survey period did not result in a noticeable increase in isolations; (5) there was no apparent correlation between campylobacter and salmonella isolations.


Deletion mutations in the small t antigen gene alter the tissue specificity of tumors induced by simian virus 40.
Subcutaneous injection of wild-type simian virus 40 into Syrian hamsters normally induces fibrosarcomas at the injection site. We showed that subcutaneous injection of three different small t deletion mutants (dl884, dl883, and dl890) led to the formation of abdominal reticulum cell sarcomas (lymphomas) in about 15% of the animals bearing tumors. The remainder of the tumors were fibrosarcomas occurring with prolonged latencies at the site of injection. We postulated that, in the absence of an active small t protein, which is thought to have cell growth-promoting properties, the mutant virus preferentially transforms rapidly proliferating lymphoid cells.


Developmental delay and multiple congenital anomalies in a child with a unique combination of partial monosomy 18 and partial trisomy 16.
A male child with multiple congenital anomalies and developmental delay is described. Cytogenetic evaluation showed that the patient was partially monosomic for the short arm of chromosome 18 and partially trisomic for the short arm of chromosome 16: a combination of chromosomal syndromes not previously described.


Head circumference, atrophy, and cognition: implications for brain reserve in Alzheimer disease.
Clinical and epidemiologic studies suggest that patients with Alzheimer disease (AD) with larger head circumference have better cognitive performance at the same level of brain pathology than subjects with smaller head circumference. A total of 270 patients with AD participating in the Multi-Institutional Research in Alzheimer's Genetic Epidemiology (MIRAGE) study underwent cognitive testing, APOE genotyping, and MRI of the brain in a cross-sectional study. Linear regression analysis was used to examine the association between cerebral atrophy, as a proxy for AD pathology, and level of cognitive function, adjusting for age, duration of AD symptoms, gender, head circumference, APOE genotype, diabetes mellitus, hypertension, major depression, and ethnicity. An interaction term between atrophy and head circumference was introduced to explore if head circumference modified the association between cerebral atrophy and cognition. There was a significant inverse association between atrophy and cognitive function, and a significant interaction between atrophy and head circumference. With greater levels of atrophy, cognition was higher for individuals with greater head circumference. This study suggests that larger head circumference is associated with less cognitive impairment in the face of cerebral atrophy. This finding supports the notion that head circumference (and presumably brain size) offers protection against AD symptoms through enhanced brain reserve.


Fast and accurate quantitative determination of the lung shunt fraction in hepatic radioembolization.
Radioembolization treatment is preceded by a 99mTc-MAA safety procedure, which is used to estimate the lung shunt fraction (LSF). Normally, the LSF is estimated by using the geometric mean of planar scintigraphy (PS-GM). However, concern has been raised about the potential overestimation of the LSF by PS-GM. Alternatively, SPECT/CT may be used for LSF estimation, but requires lengthy acquisitions, 3D segmentation, and has a limited field of view, which calls for extrapolation of the reconstructed lung counts, which introduces another source of error. We have developed a simplified SPECT/CT protocol for LSF estimation, called the quantitative orthogonal planar (QOP) method that requires only four projections to quantitatively reconstruct liver and lung activity. This mitigates the problems associated with LSF estimations from SPECT/CT. The purpose of this study was to introduce and evaluate QOP by comparing its performance to PS-GM and SPECT/CT in a retrospective patient study, and by supporting simulation experiments. Patients who received at least one 99mTc-MAA safety procedure in our center were included in this study. QOP and PS-GM were compared to SPECT/CT in Bland-Altman analyses. Supporting digital phantom experiments with a known ground-truth were performed to evaluate the performance of this method. Analysis of PS-GM versus SPECT/CT LSF estimates revealed both a larger imprecision and significant bias by PS-GM (limits of agreement: 8.1 percentage points (pp); bias: 2.7 pp). The QOP method agreed better with the SPECT/CT-based estimation (limits of agreement: 2.07 pp; bias: 0.52 pp). This observation was consistent with the digital phantom experiments. We have proposed and evaluated a novel method called QOP for LSF estimation that performs almost as accurate as SPECT/CT, but without the need for lung mass extrapolation, long scan duration, or extensive manual segmentation, making it as fast as current PS-GM.


Different dose-response relationships in the induction of different types of colonic tumors in Wistar rats by 1,2-dimethylhydrazine.
Four groups of 21 male Wistar rats were given weekly subcutaneous injections of 1,2-dimethylhydrazine at a dose of 20 mg/kg body weight for 2, 4, 6 and 8 weeks, respectively, and 7 animals of each group were killed in weeks 24, 32 and 40. This treatment induced a clear dose-related increase in the total number of colonic carcinomas. The number of tubular adenocarcinomas, which were mostly located in the descending colon, was clearly dose-related, whereas the number of signet ring cell carcinomas, which developed in the ascending colon, was apparently dose-independent.


The extent of injection site infection in injecting drug users: findings from a national surveillance study.
Injection site infections in injecting drug users (IDUs) are associated with serious morbidity and healthcare costs. Factors associated with symptoms of these were examined through annual (2006-2008) unlinked-anonymous survey of IDUs in England, Wales and Northern Ireland. Overall 36% (1863/5209) self-reported having a symptom with no trend over time (35% 2006, 37% 2007, 34% 2008). Symptoms were less common in the North East of England; increased with years injecting; and were higher in women, those recently homeless, those recently using a needle exchange, and those injecting both opiates and stimulants. Of those injecting during the previous 4 weeks (n=3733) symptoms were associated with: injecting daily; injecting >or=10 times a day; injecting into hands, groin, or legs; sharing filters; and reusing water to flush syringes. Symptoms of injection site infections are common in IDUs. Better-targeted preventive interventions are needed, and continued surveillance should assist with assessing the impact of new initiatives.


Phototoxicity of traditional chinese medicine (TCM).
Phototoxicity can cause toxic responses such as edemas and lesions, and is one of the severe adverse effects that largely limit the use of these phototoxic drugs. Some traditional Chinese medicines (TCMs) and their constituents have been reported to be phototoxic. However, to date, their phototoxicity information is still very limited, and lacks systemic investigation. This article presents the phototoxicity potential of various types of TCMs and their active components in an effort to provide valuable information for drug research and discovery to mitigate phototoxicity concerns. Some potential mechanisms of action (MoAs) of phototoxicity are discussed. In addition, in vivo and in vitro phototoxicity assays are summarized this review.


Application of a solvent-tolerant microbial consortium for biofiltration of extremely high concentration gaseous solvent streams.
The aerobic biological oxidation of 2-propanol (isopropyl alcohol, IPA) at extremely high concentrations in air by an enriched solvent-tolerant microbial consortium operating at ambient temperature was evaluated for six months. Solvent-tolerant microbial cells were immobilised onto porous glass pall rings and fed with either IPA or its metabolic product acetone as sole carbon source. Successful biofiltration of solvent vapour at a concentration of 24 g m(-3) was achieved with oxidation of up to 100% total inlet carbon. The maximum IPA mass loading and IPA elimination capacity (EC) was 1700 g m(-3) h(-1). This performance exceeds all previous values published in the literature for similar processes. A slip feed experiment, using acetone, was also performed in order to assess the substrate specificity performance. The biofilter responded successfully to a switch from acetone to IPA as sole carbon source, displaying little reduction in overall organic carbon removal.


Insulin secretion and incretin hormone concentration in women with previous gestational diabetes mellitus.
We examined the change in the levels of incretin hormone and effects of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) on insulin secretion in women with previous gestational diabetes (pGDM). A 75-g oral glucose tolerance test (OGTT) was conducted on 34 women with pGDM. In addition, 11 women with normal glucose tolerance, matched for age, height and weight, were also tested. The insulin, GIP, GLP-1, and glucagon concentrations were measured, and their anthropometric and biochemical markers were also measured. Among 34 women with pGDM, 18 had normal glucose tolerance, 13 had impaired glucose tolerance (IGT) and 1 had diabetes. No significant differences were found in GLP-1 concentration between the pGDM and control group. However, a significantly high level of glucagon was present in the pGDM group at 30 minutes into the OGTT. The GIP concentration was elevated at 30 minutes and 60 minutes in the pGDM group. With the exception of the 30-minute timepoint, women with IGT had significantly high blood glucose from 0 to 120 minutes. However, there was no significant difference in insulin or GLP-1 concentration. The GIP level was significantly high from 0 to 90 minutes in patients diagnosed with IGT. GLP-1 secretion does not differ between pGDM patients and normal women. GIP was elevated, but that does not seem to induce in increase in insulin secretion. Therefore, we conclude that other factors such as heredity and environment play important roles in the development of type 2 diabetes.


The Application of 3D Landmark-Based Geometric Morphometrics towards Refinement of the Piglet Grimace Scale.
Proper methods of assessment to objectively identify pain are essential for welfare improvements of piglets undergoing surgical castration on commercial farms. The Piglet Grimace Scale (PGS) is used to identify changes in facial expressions caused by acute pain in piglets undergoing tail docking and castration. However, subjective scoring methods are a concern for the validation of the PGS. The objectives of this study were to evaluate and refine the PGS through 3D landmark geometric morphometrics. Male piglets (n = 88) were randomly allocated to one of two treatments: castration and sham-castration. Piglet facial images were taken at four time points (before treatment, immediately post-treatment, 1 h post-treatment, and 4 h post-treatment) using a photogrammetry rig. Images were scored by four raters using five facial action units (FAUs): orbital tightening, ear position, temporal tension, lip contraction, and nose bulge/cheek tension. Three-dimensional facial models were generated and landmarked denoting 3 FAUs (orbital tightening, lip contraction, and nose bulge/cheek tension). Results suggest that orbital tightening and ear position may be reliable FAUs for the PGS. However, neither the PGS nor 3D landmark-based geometric morphometrics were able (both p > 0.10) to reliably identify facial indicators of pain in piglets undergoing castration.


Fabrication of Binder-Free Pencil-Trace Electrode for Lithium-Ion Battery: Simplicity and High Performance.
A binder-free and solvent-free pencil-trace electrode with intercalated clay particles (mainly SiO2) is prepared via a simple pencil-drawing process on grinded Cu substrate with rough surface and evaluated as an anode material for lithium-ion battery. The pencil-trace electrode exhibits a high reversible capacity of 672 mA h g(-1) at 100 mA g(-1) after 100 cycles, which can be attributed to the unique multilayered graphene particles with lateral size of few micrometers and the formation of LixSi alloys generated by interaction between Li(+) and an active Si produced in the electrochemical reduction of nano-SiO2 in the clay particles between the multilayered graphene particles. The multilayered graphene obtained by this process consists of 1 up to 20 and occasionally up to 50 sheets and thus can not only help accommodating the volume change and alleviating the structural strain during Li ion insertion and extraction but also allow rapid access of Li ions during charge-discharge cycling. Drawing with a pencil on grinded Cu substrate is not only very simple but also cost-effective and highly scalable, easily establishing graphitic circuitry through a solvent-free and binder-free approach.


Global value chains and agrifood standards: challenges and possibilities for smallholders in developing countries.
The rise of private food standards has brought forth an ongoing debate about whether they work as a barrier for smallholders and hinder poverty reduction in developing countries. This paper uses a global value chain approach to explain the relationship between value chain structure and agrifood safety and quality standards and to discuss the challenges and possibilities this entails for the upgrading of smallholders. It maps four potential value chain scenarios depending on the degree of concentration in the markets for agrifood supply (farmers and manufacturers) and demand (supermarkets and other food retailers) and discusses the impact of lead firms and key intermediaries on smallholders in different chain situations. Each scenario is illustrated with case examples. Theoretical and policy issues are discussed, along with proposals for future research in terms of industry structure, private governance, and sustainable value chains.


Facile discovery of a therapeutic agent for NK-mediated synergistic antitumor effects using a patient-derived 3D platform.
Despite the essential roles of natural killer (NK) cells in cancer treatment, the physical barrier and biological cues of the tumor microenvironment (TME) may induce NK cell dysfunction, causing their poor infiltration into tumors. The currently available two-dimensional (2D) cancer-NK co-culture systems hardly represent the characteristics of TME and are not suitable for tracking the infiltration of immune cells and assessing the efficacy of immunotherapy. This study aims to monitor NK-mediated cancer cell killing using a polymer thin film-based, 3D assay platform that contains highly tumorigenic cancer spheroids. A poly(cyclohexyl methacrylate) (pCHMA)-coated surface enables the generation of tumorigenic spheroids from pancreatic cancer patient-derived cancer cells, showing considerable amounts of extracellular matrix (ECM) proteins and cancer stem cell (CSC)-like characteristics. The 3D spheroid-based assay platform allows rapid discovery of a therapeutic agent for synergistic NK-mediated cytotoxicity through imaging-based high-content screening. In detail, the small molecule C19, known as a multi-epithelial-mesenchymal transition pathway inhibitor, is shown to enhance NK activation and infiltration via modulation of the ECM, resulting in synergistic cytotoxicity against cancer spheroids. This 3D biomimetic co-culture assay platform provides promising applications for predicting patient-specific responses to immunotherapy through advanced therapeutic combinations involving a chemical drug and immune cells.


Gravitoelectromagnetic perturbations of Kerr-Newman black holes: stability and isospectrality in the slow-rotation limit.
The most general stationary black-hole solution of Einstein-Maxwell theory in vacuum is the Kerr-Newman metric, specified by three parameters: mass M, spin J, and charge Q. Within classical general relativity, one of the most important and challenging open problems in black-hole perturbation theory is the study of gravitational and electromagnetic fields in the Kerr-Newman geometry, because of the indissoluble coupling of the perturbation functions. Here we circumvent this long-standing problem by working in the slow-rotation limit. We compute the quasinormal modes up to linear order in J for any value of Q and provide the first, fully consistent stability analysis of the Kerr-Newman metric. For scalar perturbations the quasinormal modes can be computed exactly, and we demonstrate that the method is accurate within 3% for spins J/J(max) ≲ 0.5, where J(max) is the maximum allowed spin for any value of Q. Quite remarkably, we find numerical evidence that the axial and polar sectors of the gravitoelectromagnetic perturbations are isospectral to linear order in the spin. The extension of our results to nonasymptotically flat space-times could be useful in the context of gauge-gravity dualities and string theory.


Overexpression of Insig-1 in the livers of transgenic mice inhibits SREBP processing and reduces insulin-stimulated lipogenesis.
In the current studies we generated transgenic mice that overexpress human Insig-1 in the liver under a constitutive promoter. In cultured cells Insig-1 and Insig-2 have been shown to block lipid synthesis in a cholesterol-dependent fashion by inhibiting proteolytic processing of sterol regulatory element-binding proteins (SREBPs), membrane-bound transcription factors that activate lipid synthesis. Insig's exert this action in the ER by binding SREBP cleavage-activating protein (SCAP) and preventing it from escorting SREBPs to the Golgi apparatus where the SREBPs are processed to their active forms. In the livers of Insig-1 transgenic mice, the content of all nuclear SREBPs (nSREBPs) was reduced and declined further upon feeding of dietary cholesterol. The nuclear content of the insulin-induced SREBP isoform, SREBP-1c, failed to increase to a normal extent upon refeeding on a high-carbohydrate diet. The nSREBP deficiency produced a marked reduction in the levels of mRNAs encoding enzymes required for synthesis of cholesterol, fatty acids, and triglycerides. Plasma cholesterol levels were strongly reduced, and plasma triglycerides did not exhibit their normal rise after refeeding. These results provide in vivo support for the hypothesis that nSREBPs are essential for high levels of lipid synthesis in the liver and indicate that Insig's modulate nSREBP levels by binding and retaining SCAP in the ER.


Operant ethanol self-administration after nicotine treatment and withdrawal.
The effects of chronic intermittent administration of nicotine (NIC) and withdrawal on operant ethanol (EtOH) self-administration were tested in Long-Evans rats (n = 8). EtOH self-administration (10% v/v, Fixed Ratio 4 reinforcement schedule) was induced by the sucrose-substitution procedure. Then the animals were divided into two groups of four rats matched on EtOH self-administration and the locomotor activity following an injection of NIC (0.35 mg/kg, SC) or saline was measured. The groups then received 9 days of injection of either NIC (0.35 mg/kg) or saline and then motor activity was retested using the initial NIC dose. This was followed by 17 days of NIC injections (0.6 mg/kg) or saline injections. A final locomotor test using the higher NIC dose was then conducted. The initial acute administration of NIC had no effect on motor activity compared to saline (measured by the number of horizontal movements). However, after the repeated treatment, the group of animals injected chronically and acutely with NIC showed motor activation in comparison with the animals injected chronically with saline and injected acutely with NIC only on the days of activity testing. At the end of the chronic NIC treatment, operant EtOH self-administration was not changed. However, 6 days after the NIC injections were concluded, a change in the pattern of responding for EtOH was observed in the NIC group, showing a decrease in the mean rate of responding during the first half of the operant self-administration session. When both groups were again tested for locomotor activity at the end of the operant self-administration experiment, the increased motor activity in the NIC group was still observed. The results suggest that alterations in the nicotinic system may affect EtOH self-administration, but this appears to be only modulatory, even with a significant change in locomotor response to NIC following chronic NIC administration.


Virulence characterization and molecular subtyping of typical and atypical Escherichia coli O157:H7 and O157:H(-) isolated from fecal samples and beef carcasses in Mexico.
The objective of the study was to characterize virulence genes and subtype Escherichia coli O157:H7 and O157:H( 2 ) isolates obtained from a vertically integrated feedlot slaughter plant in Mexico. A total of 1,695 samples were collected from feedlots, holding pens, colon contents, hides, and carcasses. E. coli O157:H7 detection and confirmation was carried out using conventional microbiology techniques, immunomagnetic separation, latex agglutination, and the BAX system. A total of 97 E. coli O157 strains were recovered and screened for key virulence and metabolic genes using multiplex and conventional PCR. Eighty-eight (91.72%) of the strains carried stx2, eae, and ehxA genes. Ten isolates (8.25%) were atypical sorbitol-fermenting strains, and nine were negative for the flicH7 gene and lacked eae, stx1, stx2, and ehxA. One sorbitol-positive strain carried stx2, eae, tir, toxB, and iha genes but was negative for stx1 and ehxA. Pulsed-field gel electrophoresis (PFGE) analysis yielded 49 different PFGE subtypes, showing a high genetic diversity; however, the majority of the typical isolates were closely related (80 to 90% cutoff). Atypical O157 isolates were not closely related within them or to typical E. coli O157:H7 isolates. Identical PFGE subtypes were found in samples obtained from colon contents, feedlots, holding pens, and carcasses. Isolation of a sorbitolfermenting E. coli O157 positive for a number of virulence genes is a novel finding in Mexico. These data showed that genetically similar strains of E. coli O157:H7 can be found at various stages of beef production and highlights the importance of preventing cross-contamination at the pre- and postharvest stages of processing.


Lifetime of Prevalence and Risk Factors of Problem and Pathologic Gambling in North Cyprus.
In this article, the results of the national survey of adult gambling behavior in North Cyprus (NC) in 2012 are presented. The aim of this study is to investigate the characteristics of adults' participation in gambling, and to determine the prevalence of 'problem and pathological gambling' in NC. The population of this study was formed from all the people living permanently in NC, speaking Turkish, and within the age group 18-65. Household interviews were conducted with 966 people. To obtain data, a 30 item questionnaire prepared by the researchers and a Turkish version of the Revised South Oaks Gambling Screen were used. Prevalence rates are compared with the results of the study conducted in 2007 using the same methodology and survey form. The lifetime prevalence of participating at least once in any of the 17 gambling activities investigated in the survey was 66.4 %. 3.5 % of the respondents scored as lifetime probable pathological gamblers and 9.2 % as probable problem gamblers. Risk factors for becoming probable problem and pathological gamblers include being male, being in the 19-28 age group, having a high education level, having a job and being born in Cyprus. This study shows that the prevalence of problem gambling is high in NC and increasing gradually. NC has socio-cultural features such as a history of colonization, socioeconomic problems and high unemployment, similar to other high prevalence gambling regions, which is suggestive of the importance of socio-cultural factors on gambling behavior.


Techniques for intracranial stent navigation in patients with tortuous vessels.
In some patients with stenosis of an intracranial artery, navigating the balloon or stent-delivery system is difficult of tortuous anatomy of the aortic arch, carotid arteries, or vertebral arteries Our purpose was to describe techniques of intracranial stent placement that help in navigating the stent-delivery system in tortuous vessels. Between May 1998 and June 2004, 73 symptomatic stenotic (>50%) intracranial arteries were successfully treated with stent-assisted angioplasty. In 11 cases, standard techniques of navigating the stent-delivery system into the intended lesion failed because of vascular tortuosity. In these difficult cases, several techniques were used to overcome the tortuosity. Five lesions were located in the middle cerebral arteries, four were in the supraclinoid internal carotid arteries, and two were in the distal vertebral arteries. In all difficult cases, stents were successfully placed in the intracranial artery by using several techniques: 1) waiting method in which we waited for 20-30 minutes after advancement of the microwire across the lesion, 2) the double-wire technique, and 3) the coaxial double-guiding catheter technique. The waiting method made smooth stent navigation possible in five cases, the double-wire technique was successful in four cases, and the coaxial double-guiding catheter technique was effective in two cases. No technique-related complications occurred. In difficult cases in which standard techniques of navigating the stent-delivery system into the intended lesion fail because of vascular tortuosity, our techniques were useful methods for intracranial stent navigation.


Regenerative Cells For Facial Surgery: Biofilling and Biocontouring.
Zuk et al in 2001 identified stem and regenerative cells within the stromal vascular fraction of fat. In preclinical studies, these cells appeared to stimulate angiogenesis and reduce inflammation, and soon thereafter, clinical use of stromal vascular fraction (SVF) evolved as researchers such as Rigotti, Coleman, Mojallal, our group, and others demonstrated that fat can be used for both therapeutic and aesthetic indications. The regenerative effects of fat and its contents on facial aesthetics have been shown at the histologic and cellular level. Regeneration of elastin and collagen fibers as well as improvement in capillary density and reduction of inflammation have been reported. We review our current approach to the use of regenerative cells and different types of fat grafts in facial surgery. The fat graft is classified, both from a regenerative point of view as well as a tissue product that can be modified into different tissue characteristics, depending on the area and condition treated. Clinical use of SVF enriched fat, millifat, microfat, and nanofat grafts as well as composite fat grafts are reviewed. Based on clinical experience and evidence to date, it appears that the regenerative effects seen with the use of SVF in aesthetic surgery are modest, but there appear to be definite histologic findings of regeneration. These improvements may not be clinically apparent to a patient when cell enriched fat grafts are compared to fat grafts alone. However, the subtle changes seen in histology may be cumulative over time. Three types of fat grafts are defined: millifat (parcel size 2.4<), microfat (1.2<), and nanofat (400-600 μm). Each are characterized by their injectability ratings and emulsification parcel size as well as amount of sSVF cells. Newer concepts of periosteal fat grafting, buccal fat pad grafting, pyriform aperture fat grafting, intraorbital fat grafting, and nanofat grafting are discussed. Composite fat grafts are presented as a new concept as is biofilling and biocontouring. The use of regenerative cells in facial surgery is evolving rapidly. Our understanding of the anatomic changes that occur with aging has become more precise and our ability to target histologic changes seen with aging has become more effective. Deep fat compartment grafting, superficial fat grafting, nanofat, and SVF are becoming important components of contemporary facial rejuvenation. The use of regenerative approaches in facial rejuvenation is a logical step in changing the paradigm from surgical treatment of aging to a more proactive prevention and maintenance approach that keeps up with changes in the tissues as they age.


A single skin lesion--an unusual presentation of lepromatous leprosy.
Lepromatous leprosy presenting as a solitary lesion with a high bacterial count is a rare occurrence. Such a case has been followed at the National Hansen's Disease Center, Carville, Louisiana, U.S.A., since 1977. The lesion was located on the left elbow and had been present for about nine months. The bacterial index in the lesion was 5+ and the morphological index was 0%, but slit-skin smears elsewhere were negative. The histopathology of the lesion was that of subpolar lepromatous leprosy, and the lepromin skin test was negative. Growth of the organism in the mouse foot pad was characteristic of Mycobacterium leprae, and the patient's response to dapsone monotherapy has been excellent. Therefore, this patient with a single skin lesion has lepromatous leprosy histologically, immunologically, and bacteriologically. This case illustrates the importance of slit-skin scrapings and biopsy in new cases with unusual lesions and, secondly, suggests that there are factors yet undetermined which play a significant role in determining host response to M. leprae.


Antiviral drugs for the treatment of influenza: a systematic review and economic evaluation.
To evaluate the clinical effectiveness (including adverse events) and cost-effectiveness of antivirals for the treatment of naturally acquired influenza for 'at-risk' and otherwise healthy populations. Eleven electronic databases (MEDLINE, EMBASE, Cumulative Index to Nursing and Allied Health Literature, Pascal, Science Citation Index, BIOSIS, Latin American and Caribbean Health Sciences, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effects, and Health Technology Assessment Database) were searched from October 2001 to November 2007. A supplementary search was undertaken in June 2008 for information relating to drug resistance during the 2007-8 influenza season. Systematic reviews of the evidence on the clinical effectiveness and cost-effectiveness of antivirals for the treatment of influenza were undertaken. Twenty-nine randomised controlled trials comparing antivirals with each other, placebo, or best symptomatic care were included in the evaluation of clinical effectiveness in patients presenting with an influenza-like illness (ILI). Primary outcomes were measures of symptom duration (median time to alleviation of symptoms and median time to return to normal activity). Incidence of complications, mortality, hospitalisations, antibiotic use (as a surrogate for complications) and adverse events was also assessed. In addition, an independent decision model was developed to evaluate the cost-effectiveness of antiviral treatment from the perspective of the UK NHS. Amantadine was excluded at an early stage, owing to a lack of any new trials that met the inclusion criteria and the limitations of the existing evidence. The review therefore focused on the neuraminidase inhibitors (NIs) oseltamivir and zanamivir, both of which were found to be effective in reducing symptom duration (zanamavir by 0.5-1.0 days and oseltamivir by 0.5-1.5 days). However, the effect sizes were often small and unlikely to be clinically significant in many cases, particularly in healthy adults. For the at-risk subgroups, effect sizes for differences in symptom duration were generally larger, and potentially more clinically significant, than those seen in healthy adults (median duration of symptoms reduced by 1-2 days with zanamivir and 0.50-0.75 days with oseltamivir). However, there was greater uncertainty around these results, with estimates often failing to reach statistical significance. The most consistent data and strongest evidence related to antibiotic use, with both zanamivir and oseltamivir resulting in statistically significant reductions in antibiotic use. In general, the estimates from the cost-effectiveness model were more favourable in at-risk populations (including adults and children with comorbid conditions and the elderly) compared with otherwise healthy populations. Zanamivir was the optimal NI treatment in each of the at-risk populations considered, and oseltamivir was optimal for healthy populations (both adults and children). The clinical effectiveness data for population subgroups used to inform the multiparameter evidence synthesis and cost-effectiveness modelling were, in places, limited and this should be borne in mind when interpreting the findings of this review. Trials were often not designed to determine clinical effectiveness in population subgroups and hence, although the direction of effect was clear, estimates of differences in symptom duration tended to be subject to greater uncertainty in subgroups. Despite some concerns, the use of NIs in at-risk populations appeared to be a cost-effective approach for the treatment of influenza. Well-designed observational studies might also be considered to evaluate the clinical course of influenza in terms of complications, hospitalisation, mortality and quality of life, as well as the impact of NIs.


[A simple radioimmunological method of determining thymidine in biological fluids].
A simple, sensitive and reproducible radioimmunoassay for determination of thymidine in biological fluids was developed. The technique is based on competition between 3H-thymidine and unlabelled thymidine for antithymidine antibodies. The cross-reactivity of structurally related compounds with the antibody was mostly negligible, except for deoxyuridine. The sensitivity of the assay was 10(-7) M. This radioimmunoassay may be used for measuring content of thymidine in blood serum and urine. A method for direct determination of thymidine in urine was not described before.


[Delirium in old age can be prevented and treated].
Delirium is a common neuropsychiatric syndrome characterized by disturbance of attention and consciousness developing over a short period of time. Symptoms tend to fluctuate during the course of the day. Delirium is by definition a direct physiological consequence of a general medical condition and is probably the most common presenting symptom of disease in old age. Literature review based on search on PubMed and Medline up to 31 December 2001 and a summary of several doctoral theses and our own clinical experience. Several recent intervention studies have clearly shown that delirium can be prevented and treated. Successful intervention programmes have been multifactorial and interdisciplinary, including assessment and treatment of underlying causes as well as prevention and treatment of factors endangering cerebral metabolism. In particular, cerebral hypoxemia caused by i.e. sleep-apnoea syndrome, anaemia, hypotension, pulmonary diseases, and heart failure is often easily prevented and treated. Excellent nursing care seems to be a prerequisite for successful prevention and treatment of delirium. Acceptable scientific evidence for pharmacological treatment is still lacking; it is sometimes necessary but should be used with caution. If sedation is acceptable, clomethiazole is the drug of choice but if the delirium is complicated by frightening hallucinations and agitation, haloperidol or risperidone can be used but only for short periods. Cholinesterase inhibitors are probably a better choice, though randomised treatment studies are still lacking.


Genome-wide expression analysis of salt-stressed diploid and autotetraploid Paulownia tomentosa.
Paulownia tomentosa is a fast-growing tree species with multiple uses. It is grown worldwide, but is native to China, where it is widely cultivated in saline regions. We previously confirmed that autotetraploid P. tomentosa plants are more stress-tolerant than the diploid plants. However, the molecular mechanism underlying P. tomentosa salinity tolerance has not been fully characterized. Using the complete Paulownia fortunei genome as a reference, we applied next-generation RNA-sequencing technology to analyze the effects of salt stress on diploid and autotetraploid P. tomentosa plants. We generated 175 million clean reads and identified 15,873 differentially expressed genes (DEGs) from four P. tomentosa libraries (two diploid and two autotetraploid). Functional annotations of the differentially expressed genes using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed that plant hormone signal transduction and photosynthetic activities are vital for plant responses to high-salt conditions. We also identified several transcription factors, including members of the AP2/EREBP, bHLH, MYB, and NAC families. Quantitative real-time PCR analysis validated the expression patterns of eight differentially expressed genes. Our findings and the generated transcriptome data may help to accelerate the genetic improvement of cultivated P. tomentosa and other plant species for enhanced growth in saline soils.


Vasoactive factors in sickle cell disease: in vitro evidence for endothelin-1-mediated vasoconstriction.
While systemic plasma endothelin-1 (ET-1) levels are increased during acute crisis in sickle cell disease, the relative levels of potent vasoactive factors that contribute to the regulation of vascular function, such as ET-1, NO, and cell-free hemoglobin, during the course of periodic vaso-occlusive episodes remain unclear. Moreover, whether and to what extent sickling-induced release of ET-1 alters vascular tone is not completely understood. To investigate the sequential changes in circulating vasoactive factors, we measured plasma ET-1, NO metabolites (NOx), and cell-free hemoglobin (Hb) before (steady-state), during (crisis), and after a vaso-occlusive (post-crisis) episode. Steady-state ET-1 levels (fmol/mL) increased from 2.3 +/- 0.4 to 11.0 +/- 1.4 and 4.2 +/- 1.0 during crisis and post-crisis periods, respectively. There was no significant difference in plasma NOx levels. Cell-free Hb levels were significantly higher in sickle cell patients in all phases as compared to the control group, and especially during crisis cell-free Hb levels were elevated by 4-fold (209,000 +/- 31,000 vs. 46,000 +/- 5,300 ng/mL in steady-state). Conditioned medium from human pulmonary artery endothelial cells exposed to sickled erythrocytes prepared by deoxygenation induced contraction of aortic rings, and this effect was blocked by an ET(A) receptor antagonist. These findings indicate that ET-1 is the predominant contractile factor released by cultured endothelial cells upon exposure to deoxygenated sickled SS erythrocytes and ET-1-NO-NO scavenger balance is altered in favor of vasoconstriction during an acute episode in SCD.


Clinical evaluation of marketed orthodontic products: are researchers behind the times? A meta-epidemiological study.
The role of marketing and industry in the treatment decisions of orthodontists has received increasing attention in recent years with clinical research typically undertaken subsequent to established use of these devices and often failing to confirm the promise of manufacturers' claims. This meta-epidemiological study was undertaken to assess the proportion of clinical trials in orthodontics evaluating commercially marketed products and to evaluate the direction of the results of these studies. Electronic searching was undertaken to identify randomized controlled trials (RCTs) published over a 5-year period (1 January 2012 to 31 December 2016). Data obtained included the type of marketed intervention, direction of effect and declaration of both industry sponsorship and conflict of interest. Eighty-four RCTs published in 23 scientific journals were included with the highest percentage in the American Journal of Dentofacial Orthopedics (AJO-DO) (23.8%), followed by the European Journal of Orthodontics (EJO) (14.3%), Journal of Orthodontics (JO) (10.7%) and Angle Orthodontist (AO) (10.7%). Overall, 45% (38/84) of clinical trials assessed involved analysis of marketed products after their introduction. Interventions to improve oral health or circumvent the risk of iatrogenic damage, such as white spot lesions, were most commonly assessed (15.8%), with the relative merits of non-surgical adjuncts (14.1%) and other orthodontic auxiliaries (13.1%) also frequently evaluated. In 44% of RCTs, a positive effect of the marketed intervention was not reported. Industry sponsorship of the research was declared in 9.5% RCTs. No significant associations between the direction of the effect and both declaration of industry sponsorship (p = 0.56) and conflict of interest (p = 0.96) were detected. Moreover, for marketed and non-marketed products, no significant associations for both declaration of industry sponsorship (p = 0.44) and conflict of interest (p = 0.28) were found. Almost half of orthodontic clinical trials over the past 5 years involve analysis of marketed products after their introduction. The results highlight a potential source of waste in orthodontic research emanating from existing approaches to licensing and marketing of orthodontic products.


Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology.
Heavy metal pollution has become an environmental problem throughout the world because heavy metal can be accumulated into the food chain and bring about serious problems, not only for ecosystems but also for human health. In this study, functional metal-binding proteins (FMBPs) were isolated from a metal-stimulated activated sludge culture with the aim of applying them to an innovative metal removal technology. Activated sludge bacteria was cultured in growth media including copper ion, and the stimulation of protein production by copper ion led to the 14% increase in a quantity of extracted crude proteins per 1 g of bacterial cell pellet (wet). In order to isolate FMBPs, extracted crude proteins were applied to the immobilized metal affinity column in which each of copper, nickel and zinc was used as a ligand. Several FMBPs were succesfully isolated from copper-stimulated bacteria. One of FMBPs (molecular weight of about 40 kDa) exhibited an ability to adsorb all three metals. The multi metal-binding property of this FMBP could be applied to an innovative metal removal technology. Furthermore, isolated FMBPs that could capture only one kind of heavy metal would also be attractive as a metal adsorbent in recovering a specific metal as a resource from wastewater, including several heavy metals.


Lymphosarcoma cell leukemia: the contribution of cell surface study to diagnosis.
Cell surface immunoglobulin, complement receptor, and spontaneous rosette formation with sheep erythrocytes were investigated in 43 patients with malignant lymphoma, including 13 with lymphosarcoma cell leukemia, and in 59 patients with chronic lymphocytic leukemia. The quantity of immunoglobulin on the lymphocyte surface was estimated from the intensity of fluorescent staining with fluorescein-conjugated anti-immunoglobulin antisera. At least two, and probably three, B cell species could be recognized by cell surface study. Cells from chronic lymphocytic leukemia and diffuse well-differentiated lymphocytic lymphoma had sparse amounts of surface immunoglobulin, while the cells of diffuse poorly differentiated lymphocytic lymphoma had large quantities of this material. Nodular lymphoma probably represented a third B-cell subtype with intermediate amounts of surface immunoglobulin. The lymphocytes of chronic lymphosarcoma cell leukemia exhibited the intense surface staining, which was characteristic of the underlying poorly differentiated lymphocytic lymphoma (diffuse or nodular), and could be readily distinguished from the faint-staining chronic lymphocytic leukemia cells.


Examining rural-urban disparities in perceived need for health care services among adults with disabilities.
The purpose of this study is to parse out differences between unmet need and perceived need for health care services among rural and urban adults with disabilities in the United States. While unmet need focuses primarily on environmental factors such as access to health insurance or provider availability, perceived need relates to personal choice. This distinction between unmet and perceived need is largely ignored in prior studies, but relevant to public health strategies to improve access and uptake of preventive care. Using Wave 2 data from the National Survey on Health and Disability, we explored rural and urban differences in unmet and perceived health care needs among working-age adults with disabilities for acute and preventive services. Although we found no significant differences in unmet needs between rural and urban respondents, we found that perceived needs for dental care and mental health counseling varied significantly across geography. Using logistic regression analysis and controlling for observable participant characteristics, we found that respondents living in noncore counties relative to metropolitan counties were more likely to report not needing dental care (OR 1.89, p = 0.028), and not needing mental health counseling services (OR 2.15, p ≤ 0.001). These findings suggest additional study is warranted to understand perceived need for preventive services and the levers for addressing rural disparities.


Vitamin D status associates with skeletal muscle loss after anterior cruciate ligament reconstruction.
BACKGROUNDAlthough 25-hydroxyvitamin D [25(OH)D] concentrations of 30 ng/mL or higher are known to reduce injury risk and boost strength, the influence on anterior cruciate ligament reconstruction (ACLR) outcomes remains unexamined. This study aimed to define the vitamin D signaling response to ACLR, assess the relationship between vitamin D status and muscle fiber cross-sectional area (CSA) and bone density outcomes, and discover vitamin D receptor (VDR) targets after ACLR.METHODSTwenty-one young, healthy, physically active participants with recent ACL tears were enrolled (17.8 ± 3.2 years, BMI 26.0 ± 3.5 kg/m2). Data were collected through blood samples, vastus lateralis biopsies, dual energy x-ray bone density measurements, and isokinetic dynamometer measures at baseline, 1 week, 4 months, and 6 months after ACLR. The biopsies facilitated CSA, Western blotting, RNA-seq, and VDR ChIP-seq analyses.RESULTSACLR surgery led to decreased circulating bioactive vitamin D and increased VDR and activating enzyme expression in skeletal muscle 1 week after ACLR. Participants with less than 30 ng/mL 25(OH)D levels (n = 13) displayed more significant quadriceps fiber CSA loss 1 week and 4 months after ACLR than those with 30 ng/mL or higher (n = 8; P < 0.01 for post hoc comparisons; P = 0.041 for time × vitamin D status interaction). RNA-seq and ChIP-seq data integration revealed genes associated with energy metabolism and skeletal muscle recovery, potentially mediating the impact of vitamin D status on ACLR recovery. No difference in bone mineral density losses between groups was observed.CONCLUSIONCorrecting vitamin D status prior to ACLR may aid in preserving skeletal muscle during recovery.FUNDINGNIH grants R01AR072061, R01AR071398-04S1, and K99AR081367.


Clinical features and hMSH2/hMLH1 germ-line mutations in Chinese patients with hereditary nonpolyposis colorectal cancer.
At least five mismatch repair (MMR) genes, including hMSH2, hMLH1, hPMS, hPMS2, and hMSH6/GTBP, are associated with hereditary nonpolyposis colorectal cancer (HNPCC). More than 90% of families with HNPCC harbor the hMSH2 and hMLH1 gene mutations. We have analyzed the clinical features of HNPCC among Chinese patients and report the results of screening for mutations in the hMSH2 and hMLH1 genes. The data concerning gender, site of colorectal cancer (CRC), age at diagnosis, history of synchronous and/or metachronous colorectal cancer, instance of extracolonic cancers, and histopathology of tumors for 126 patients from 28 independent families with HNPCC were collected. Fifteen of the families met the Amsterdam I criteria, and 13 met the Japanese clinical criteria for diagnosis. Genomic DNA was extracted from the peripheral lymphocytes. Polymerase chain reaction (PCR) and denaturing high-performance liquid chromatography (DHPLC) were used to screen the coding region of the hMSH2 and hMLH1 genes. Samples showing abnormal DHPLC profiles were sequenced. One hundred and seventy malignant neoplasms were found in the 126 patients, of whom 23 had multiple cancers. Ninety-eight of the patients (77.8%) had colorectal cancers, with an average age at onset of 45.9 years and a right-sided predominance. Eight hMSH2 or hMLH1 gene sequence variations were found in 12 families, and a germ-line G204X nonsense mutation in the third exon of hMSH2 was found, representing the first mutation in an MMR gene ever found in people of Chinese Mongolian ethnicity. HNPCC is a typical autosomally dominant hereditary disease, characterized by early onset, a predominance of proximal colorectal cancer, and multiple synchronous and metachronous colorectal cancers. DHPLC is a powerful tool for detecting mutations in the hMSH2 and hMLH1 genes. Mutations in the first nine exons of the hMLH1 gene were more common in Chinese patients.


Structures of apo IRF-3 and IRF-7 DNA binding domains: effect of loop L1 on DNA binding.
Interferon regulatory factors IRF-3 and IRF-7 are transcription factors essential in the activation of interferon-β (IFN-β) gene in response to viral infections. Although, both proteins recognize the same consensus IRF binding site AANNGAAA, they have distinct DNA binding preferences for sites in vivo. The X-ray structures of IRF-3 and IRF-7 DNA binding domains (DBDs) bound to IFN-β promoter elements revealed flexibility in the loops (L1-L3) and the residues that make contacts with the target sequence. To characterize the conformational changes that occur on DNA binding and how they differ between IRF family members, we have solved the X-ray structures of IRF-3 and IRF-7 DBDs in the absence of DNA. We found that loop L1, carrying the conserved histidine that interacts with the DNA minor groove, is disordered in apo IRF-3 but is ordered in apo IRF-7. This is reflected in differences in DNA binding affinities when the conserved histidine in loop L1 is mutated to alanine in the two proteins. The stability of loop L1 in IRF-7 derives from a unique combination of hydrophobic residues that pack against the protein core. Together, our data show that differences in flexibility of loop L1 are an important determinant of differential IRF-DNA binding.


DNA sequence analysis of crown gall tumor T-DNA encoding the 0.7 kb transcript.
Crown gall tumor formation involves integration into the plant genome of DNA sequences (the T-region) of tumor-inducing (Ti) plasmids present in Agrobacterium tumefaciens. The T-DNA of the tumor expresses several gene products. Little is known about the function or regulation of expression of the 0.7kb transcript, which represents a relatively abundant T-DNA transcript in octopine-type tumors. In this report, a detailed structural analysis of the gene encoding the 0.7 kb transcript has been obtained by DNA sequence analysis of T-DNA isolated from A6S/2 tumor line. An indication of the structural characteristics of the protein product is obtained from the predicted amino acid sequence. The sequences flanking the open reading frame show characteristics with other eucaryotic genes. The corresponding DNA sequence of the inducing Ti plasmid (pTiA6) is identical with that of the DNA sequence from the tumor. Comparison of this gene sequence with the corresponding region of another Ti plasmid (pTiAch5) shows several differences in the 5' flanking sequence, but the nucleotide sequence of the coding region and 3' flanking region are identical.


Gene-specific differences in the supranucleosomal organization of rat liver chromatin.
Rat liver chromatin was separated into a solubilized portion and insoluble nuclear material, and the solubilized portion was fractionated by sucrose gradient sedimentation. The chromatin encompassing three transcribed genes (albumin, tryptophan oxygenase, and alpha-fetoprotein), which are expressed at very different levels, partitions preferentially with insoluble nuclear material and possesses a disrupted nucleosome structure. On the contrary, the chromatin encompassing three inactive genes fractionates into the solubilized chromatin portion and exhibits a canonical nucleosome repeat structure. By sucrose gradient sedimentation, all size classes of inactive chromatin particles are found to contain internal cleavages in the linker region between nucleosomes; they are probably held together by histone H1 and mono- and divalent cations. When the chromatin encompassing two flanking sequences of the tyrosine aminotransferase gene is studied, the 0.35-kilobase upstream-located chromatin exhibits features of active genes, while the 2.55-kilobase upstream-located chromatin partitions preferentially with solubilized chromatin and sediments in internally cleaved particles.


Relationship between bone adaptation and in-vivo mechanical stimulus in biological reconstructions after bone tumor: A biomechanical modeling analysis.
Biomechanical interpretations of bone adaptation in biological reconstructions following bone tumors would be crucial for orthopedic oncologists, particularly if based on quantitative observations. This would help plan for surgical treatments, rehabilitative programs and communication with the patients. We aimed to analyze the biomechanical adaptation of a femoral reconstruction after Ewing sarcoma according to an increasingly-used surgical technique, and to relate in-progress bone resorption to the mechanical stimulus induced by different motor activities. We created a multiscale musculoskeletal and finite element model from CT scans and motion analysis data at a 76-month follow-up of a patient, to analyze muscle and joint loads, and to compare the mechanical competence of the reconstructed bone with the contralateral limb, in the current real condition and in a possible revision surgery that removed proximal screws. Our results showed strategies of muscle coordination that led to differences in joint loads between limbs more marked in more demanding motor activities, and generally larger in the contralateral limb. The operated femur presented a markedly low ratio of physiological strain due to load-sharing with the metal implant, particularly in the lateral aspect. The possible revision surgery would help restore a physiological strain configuration, while the safety of the reconstruction would not be threatened. We suggest that bone resorption is related to load-sharing and to the internal forces exerted during movement, and the mechanical stimulus should be improved by adopting modifications in the surgical treatment and by promoting physical therapy aimed at specific muscle strengthening.


Exploding-wire experiments and theory for metal conductivity evaluation in the sub-eV regime.
Copper and silver wires are subjected to pulsed high current densities producing high density metal plasma in the sub-eV regime with atmospheric air as a background gas. Numerical simulation via application of the one-dimensional magnetohydrodynamic partial differential equations solved simultaneously with the constraining circuit equations is presented. The simulations require accurate knowledge of the material equation of state (EOS) and transport properties; the LANL sesame database is applied for the EOS in all cases. Two electrical conductivity models are applied. First, the Lee-More-Desjarlais (LMD) and its modification, the quantum LMD (QLMD) conductivity, which have been well proven at higher temperatures, are applied. Simulations with the LMD and QLMD data indicate that the conductivity data as well as the MHD methodology are accurate in the sub-eV regime of interest. A less computationally involved, empirical conductivity model is applied in the same regime to explore its temperature-density range of applicability compared to the more sophisticated model.


Cyr61 protects against hyperoxia-induced cell death via Akt pathway in pulmonary epithelial cells.
We have used gene expression profiling approaches to identify new molecular targets in various models of lung injury and human lung diseases. Among the many genes that are significantly induced in these studies, cysteine-rich61 (Cyr61) consistently ranks as one of the most significant genes. Here, we use the well-established model of hyperoxia to better understand the function of Cyr61 in acute lung injury. Cyr61, a stress-related immediate-early response gene, has known diverse functions involving angiogenesis, tumorigenesis, and wound repair. It belongs to the newly discovered "CCN" family containing six growth and regulatory factors. We showed that hyperoxia induces Cyr61 expression in a variety of pulmonary cells and in lung tissue in vivo. Loss of function studies, by suppressing Cyr61 expression by siRNA, accelerated lung epithelial cell death after hyperoxia. Gain of function studies, by overexpressing Cyr61, significantly conferred increased resistance to hyperoxia-induced cell death. Moreover, cells overexpressing Cyr61 induce Akt activation. Inhibition of Akt by siRNA abrogated the protective effects of Cyr61-overexpressing cells in response to hyperoxia. Taken together, our data demonstrate that Cyr61 expression provides cytoprotection in hyperoxia-induced pulmonary epithelial cell death and that this effect was in part mediated via the Akt signaling pathway.


Formation of malondialdehyde adducts in livers of rats exposed to ethanol: role in ethanol-mediated inhibition of cytochrome c oxidase.
Previous studies in our laboratory demonstrated that short-term ethanol consumption by maternal rats increased the hepatic levels of 4-hydroxynonenal (HNE) in both the adult and the fetus. Additionally, HNE inhibited cytochrome c oxidase (COX) by forming adducts with the enzyme subunits. The present study examined modification of COX by another major aldehydic lipid peroxidation product, malondialdehyde (MDA), and its role in COX inhibition by ethanol. It is demonstrated in vitro that MDA inhibits the activity of purified COX while forming adducts with the enzyme. Compared with HNE, MDA is a more potent inhibitor of COX. Overnight incubation at room temperature caused an 80% decrease in COX activity by MDA versus a 67% decrease by HNE. MDA produced marked inhibition of COX activity at physiologically relevant concentrations, e.g., 43% inhibition at 10 microM. Although our previous studies documented that HNE formed adducts primarily with subunit IV of COX via histidine residues, the current report showed that MDA forms adducts with both subunit IV and subunit V via lysine residues. Furthermore, both aldehydes induce carbonyl formation in subunit IV. The in vivo role of MDA in the impairment of COX by ethanol is assessed in both adult and fetal liver after maternal ethanol consumption. The results showed that: (1) there are significant increases in MDA levels in liver homogenate as well as mitochondria in both adult and fetal livers after ethanol exposure; (2) these MDA levels are in the nanomole/mg protein range, in contrast to picomole/mg protein range of HNE in identical setting; and (3) ethanol-induced production of MDA is accompanied by enhanced formation of MDA adducts with COX. These findings suggest that MDA may play at least as equally an important role as HNE in ethanol-induced inhibition of COX.


Diurnal potassium excretory cycles in the rat.
Diurnal potassium cycles (DPC) were measured in unanesthetized undisturbed rats fed a liquid diet and maintained in a 12-h light-dark environment. A fourfold step increase in diet potassium content increased DPC amplitude without altering phase. After presentation of the high-potassium diet, the initial adaptive increase in excretion occurred within 1.5 h (diet given during dark phase) and within 6 h (diet given during light phase). On a day when food was withheld (no potassium intake), DPC were present but with a lowered amplitude. The amount of potassium excreted on a fasting day exceeded gut and extracellular fluid potassium content and was only modestly increased when rats were previously fed a high-potassium diet. In adrenalectomized rats that received no steroid replacement or received constant infusions of low levels of aldosterone, dexamethasone, or aldosterone plus dexamethasone, potassium balance and DPC were normal. It is concluded that the amplitude of DPC in the rat is determined in part by the availability of potassium from both intracellular and extracellular potassium pools; mechanisms independent of potassium intake can generate the DPC; and the presence or the cyclic secretion of adrenal steroids is not necessary for the generation of DPC in the rat.


Alcohol expectancies among male alcoholics, problem drinkers, and nonproblem drinkers.
Drinkers' expectancies regarding alcohol effects are cognitive factors hypothesized to affect decisions to drink and responses to alcohol. Recent research in this area has yielded descriptive data mostly on nonalcoholic drinkers' motives for drinking and alcohol expectancies. Much less attention has been focused on assessing the extent to which these findings generalize to a broader array of drinkers. Alcohol expectancies were assessed in this report among 260 alcoholics, 79 problem drinkers, and 81 nonproblem drinkers (all subjects were male) using a comprehensive alcohol expectancy measure. Alcoholics almost uniformly scored higher than the problem and nonproblem drinkers in their endorsements of alcohol's effects in different domains (e.g., alcohol acts as a global positive transforming agent, enhances physical and social pleasure, engenders assertiveness, reduces tension). Problem drinkers also tended to score higher than the nonproblem drinkers in these domains. A multiple discriminant analysis yielded two functions which together were moderately successful in classifying these subjects according to drinker group. Taken together, these data suggest that important individual differences exist in drinkers' expectations regarding alcohol's utility, and that alcohol expectancies vary considerably as a function of problem drinking severity. The clinical implications of these findings are discussed.


Membrane integration into PDMS-free microfluidic platforms for organ-on-chip and analytical chemistry applications.
Membranes play a crucial role in many microfluidic systems, enabling versatile applications in highly diverse research fields. However, the tight and robust integration of membranes into microfluidic systems requires complex fabrication processes. Most integration approaches, so far, rely on polydimethylsiloxane (PDMS) as base material for the microfluidic chips. Several limitations of PDMS have resulted in the transition of many microfluidic approaches to PDMS-free systems using alternative materials such as thermoplastics. To integrate membranes in those PDMS-free systems, novel alternative approaches are required. This review provides an introduction into microfluidic systems applying membrane technology for analytical systems and organ-on-chip as well as a comprehensive overview of methods for the integration of membranes into PDMS-free systems. The overview and examples will provide a valuable resource and starting point for any researcher that is aiming at implementing membranes in microfluidic systems without using PDMS.


Fibroblast growth factor 23: a possible cause of left ventricular hypertrophy in hemodialysis patients.
Left ventricular hypertrophy (LVH) is a common cardiovascular disorder and an independent risk factor for cardiovascular death in dialysis patients. Hyperphosphatemia is associated with LVH. Previous studies have shown that fibroblast growth factor 23 (FGF23), which has an important role in phosphate metabolism, is elevated in chronic hemodialysis patients. The aim of this study is to determine the association of FGF23 and LVH and the prognostic value of FGF23 in chronic hemodialysis patients. One hundred twenty-four end-stage renal disease patients were evaluated for LVH by echocardiography. Serum FGF23 levels were measured using a commercial enzyme-linked immunosorbent assay kit. Patients with LVH were more likely to have poor urea clearance (Kt/V), higher systolic blood pressure, and comorbidity of diabetes mellitus and coronary artery disease. LVH was also associated with higher levels of FGF23. Multivariate analysis indicated that FGF23 level, systolic blood pressure, and comorbidity of diabetes mellitus and coronary artery disease remained correlated with LVH. This suggested that serum FGF23 level is independently associated with LVH in our hemodialysis patients. Cox analysis indicated no significant difference in risk of death for patients with elevated levels of FGF23. LVH has a high prevalence in hemodialysis patients, and FGF23 is independently associated with LVH but is not a predictor for short-term prognosis (2-year follow-up).


It's Your Game…Keep It Real: can innovative public health prevention research thrive within a comparative effectiveness research framework?
The federal comparative effectiveness research (CER) initiative is designed to evaluate best practices in health care settings where they can be disseminated for immediate benefit to patients. The CER strategic framework comprises four categories (research, human and scientific capital, data infrastructure, and dissemination) with three crosscutting themes (conditions, patient populations, and types of intervention). The challenge for the field of public health has been accommodating the CER framework within prevention research. Applying a medicine-based, research-to-practice CER approach to public health prevention research has raised concerns regarding definitions of acceptable evidence (an evidence challenge), effective intervention dissemination within heterogeneous communities (a dissemination and implementation challenge), and rewards for best practice at the cost of other promising but high-risk approaches (an innovation challenge). Herein, a dynamic operationalization of the CER framework is described that is compatible with the development, evaluation, and dissemination of innovative public health prevention interventions. An effective HIV, STI, and pregnancy prevention program, It's Your Game…Keep It Real, provides a case study of this application, providing support that the CER framework can compatibly coexist with innovative, community-based public health prevention research.


In vitro synthesis of IgG by peripheral blood lymphocytes in chronic liver disease.
In vitro IgG synthesis by peripheral blood mononuclear cells (PBM) from patients with chronic liver disease (CLD) was studied. In addition, the effect of pokeweed mitogen (PWM), polyadenylic-polyuridylic acid complexes (poly A:U) and thymosin fraction 5 on IgG synthesis was determined. Unstimulated cultures of PBM from patients with chronic active hepatitis (CAH) and alcoholic cirrhosis (AC) synthesized significantly higher quantities of IgG than the controls. Moreover, there was a direct correlation between serum IgG concentrations and the quantity of newly synthesized IgG in these unstimulated cultures. PWM, poly A:U and thymosin each stimulated increased IgG synthesis in the controls. While neither poly A:U nor thymosin enhanced IgG synthesis in patients with CLD, PWM increased IgG synthesis in CAH but not AC. These results indicate that spontaneous in vitro B cell synthesis of IgG is enhanced in CLD and may reflect antigenic stimulation in vivo.


Cell cycle-related phosphatases CDC25A and B expression correlates with survival in ovarian cancer patients.
Alterations in cell cycle regulating proteins are common in many cancer types. Recent data suggest a possible link between CDC25 phosphatases overexpression and malignancy. Our objective was to evaluate the expression of the three cell cycle-related phosphatases CDC25A, CDC25B and CDC25C in patients with ovarian cancer. All the patients had a minimal follow up of three years. CDC25A, CDC25B and CDC25C expression were investigated by immunohistochemistry in 106 patients with ovarian cancer. CDC25A and CDC25B were found expressed in almost all the samples analyzed, while CDC25C was undetectable in more than 80% of the patients. The low evaluable data on CDC25C expression, did not allow any association between the expression of this phosphatase and prognosis. The expression of CDC25A and CDC25B showed some evidences of association with a poor prognosis (p = 0.034 and p = 0.058 respectively). This relationship was independent of other factors such as tumor grade, histotype, stage and residual tumor after surgery. In the same patients the examined parameters (residual tumor, grade, stage and histotype) did show the expected relation with survival. The results indicate that high CDC25A and CDC25B expression is related to a worse prognosis in ovarian cancer patients. CDC25 phosphatases expression can be regarded as a possible prognostic factor for ovarian cancer and these proteins should be evaluated as potential molecular targets of novel drugs against this human neoplasm.


Psychosocial treatment of posttraumatic stress disorder.
This article reviews empirically validated psychosocial treatments for posttraumatic stress disorder (PTSD) and considers factors associated with successful therapy outcome. Most of the treatments whose efficacy was studied empirically fall within the broad category of cognitive-behavioral therapy. These include exposure therapy, anxiety management programs, and cognitive therapy. These therapy modalities have been developed to modify conditioned fear and erroneous cognitions that are thought to underlie PTSD. Exposure therapy has the most empirical support because it was found to be effective across different populations of trauma victims with PTSD. Combinations of therapies have also been used, and the value of these is discussed. In addition, this article presents recent evidence about the efficacy of eye movement and desensitization reprocessing. A growing body of evidence supports the use of psychosocial treatments for PTSD, but not all patients benefit. Future research should develop programs that increase the motivation of patients to take advantage of these efficacious treatments.


Increased plasma triglycerides, cholesterol and apolipoprotein E during prolonged fasting in normal subjects.
Plasma lipid and high density lipoprotein (HDL) levels were studied in 20 normal, healthy, non-obese males while fasting (150 kcal/d with free intake of water) for 6 d in a hunger strike. Plasma triglyceride and cholesterol levels were increased by 18% after 6 d of fasting. HDL-cholesterol concentration was not significantly changed for 4 d, but decreased by 22% after 6 d. Platelet aggregation induced by adenosine diphosphate (ADP) or collagen after 6 d of fasting was in the normal range. In 3 subjects fasted for 9 d, a complete plasma lipoprotein analysis was done. Very low and low density lipoprotein (VLDL and LDL) levels were elevated, whereas HDL was reduced after 9 d of fasting. On isoelectric focusing analysis, a marked reduction in apolipoprotein (apo) E concentration in both VLDL and HDL was noted. Liver function tests showed a reduction in hepatic enzyme activity; and since apo E is of hepatic origin also, we suggest that long fasting inhibits liver function in normal subjects.


Mitochondrial DNA mutations as a fundamental mechanism in physiological declines associated with aging.
The hypothesis that mitochondrial DNA damage accumulates and contributes to aging was proposed decades ago. Only recently have technological advancements, which facilitate microanalysis of single cells or portions of cells, revealed that mtDNA deletion mutations and, perhaps, single nucleotide mutations accumulate to physiologically relevant levels in the tissues of various species with age. Although a link between single nucleotide mutations and physiological consequences in aging tissue has not been established, the accumulation of deletion mutations in skeletal muscle fibres has been associated with sarcopenia. Different, and apparently random, deletion mutations are specific to individual fibres. However, the mtDNA deletion mutation within a phenotypically abnormal region of a fibre is the same, suggesting a selection, amplification and clonal expansion of the initial deletion mutation. mtDNA deletion mutations within a muscle fibre are associated with specific electron transport system abnormalities, muscle fibre atrophy and fibre breakage. These data point to a causal relationship between mitochondrial DNA mutations and the age-related loss of muscle mass.


Reassessment of the taxonomic structure of the diazotrophic genus Azoarcus sensu lato and description of three new genera and new species, Azovibrio restrictus gen. nov., sp. nov., Azospira oryzae gen. nov., sp. nov. and Azonexus fungiphilus gen. nov., sp. nov.
The taxonomic structure of members of the genus Azoarcus sensu lato was reassessed in a polyphasic approach. Two species, Azoarcus communis and Azoarcus indigens, three unnamed species containing diazotrophs associated with Kallar grass roots (groups C, D) and a group of strains (E) isolated from fungi were analysed. They were compared by PAGE analyses of cellular proteins, genomic fingerprints, morphological and nutritional features to new isolates from rice roots. All strains within groups C, D and E containing 5-12 isolates showed group-specific cell and colony morphology and carbon source utilization patterns, with exception of the obligately microaerobic strain BS20-3, a member of group C. All strains, with this exception, also had almost indistinguishable electrophoretic protein patterns and genomic fingerprints generated with tDNA-directed primers, suggesting they belong to the same species. Phylogenetic analyses of almost complete 16S rDNA sequences carried out with three different algorithms (neighbour-joining, maximum-likelihood, parsimony) revealed that Azoarcus sensu lato is not monophyletic. Groups C, D and E formed three distinct lineages located between the Azoarcus/Thauera and the Rhodocyclus clusters. Phylogenetic distances between groups C, D and E were as large as between other genera (93-94% sequence similarity). This suggested they have the rank of three different genera. Since it was possible to differentiate them from each other and other related bacteria by phenotypic features, three new genera with one type species each are proposed: Azovibrio restrictus gen. nov., sp. nov., Azospira oryzae gen. nov., sp. nov. and Azonexus fungiphilus gen. nov., sp. nov.


[New approaches in the treatment of high intra-articular fractures of the mandibular condylar process].
This paper presents treatment experience obtained upon 7 patients studied case of mandible condylar process high intra-articular fractures accompanied by the dislocation of the articular head. To reach dislocated head the osteotomy of hinder border ramus mandible together with the condylar process neck was carried out. And the latter was rewomed form the wound. After that the extraction of all the head itself followed. A side the wound the osteosynthesis with the help of titanium bone screws and miniplates was conducted. Then the fragment of ascending ramus mandible with attached mandible condylar condylar process head was replanted into the wound. High immediate and late results (followed up for 18 months) demonstrate the potential efficiency of the proposed method.