Self-assembled pH-responsive films prepared from mussel anchoring threads.
The byssus is a series of collagen-rich fibers securing mussels to surfaces. The complex but elegant heterogeneous assembly of the various proteins in the threads is responsible for their remarkable mechanical properties combining strength and extensibility. Along with the well-known biocompatibility and biodegradability attributed to collagen-based materials, these mechanical properties are highly desirable to produce biomaterials for soft tissue engineering and drug delivery applications. In order to replicate the byssus natural features and properties, we prepared a soluble byssus protein hydrolyzate (BPH) that can generate water-insoluble self-standing films. Atomic force and scanning electron microscopy revealed the presence of self-assembled collagen-like fibrils at the surface of the films. Infrared spectroscopy analysis of the film formation showed that insolubility is caused by the self-assembly of polypeptides from the hydrolyzate into antiparallel β-sheets, aggregated β-strands and collagen triple-helix structures. The mechanical properties and water swelling measurements on the films can be reversibly pH-modulated by modifying the electrostatic interactions between the ∼30 mol% of charged amino acids. Optimal mechanical properties and minimum swelling are obtained at the isoelectric point (pH 4.5). Higher or lower pH treatment reversibly decreases their stiffness and strength and increases their swelling ratio. Altogether, our results show that byssus proteins are an interesting sustainable feedstock for preparing new solid-state pH-tuneable biomaterials.


Build-up cap materials for measurement of photon head-scatter factors.
The suitability of high-Z materials as build-up caps for head-scatter measurements has been investigated. Build-up caps are often used to enable characterization of fields too small for a mini-phantom. We have studied lead and brass build-up caps with sufficiently large wall thicknesses, as compared to the range of contaminating electrons originating in the accelerator head, and compared them with build-up caps made of ionization chamber equivalent materials, i.e. graphite. The results were also compared with measurements taken using square and cylindrical polystyrene mini-phantoms. Field sizes ranging from 3 cm x 3 cm up to 40 cm x 40 cm were studied for nominal photon energies of 4, 6, 10 and 18 MV. The results show that the use of lead and brass build-up caps produces normalized head-scatter data slightly different from graphite build-up caps for large fields at high photon energies. At lower energies, however, no significant differences were found. The intercomparison between the two different plastic mini-phantoms and graphite caps showed no differences.


Direct Pulmonary Vein Ablation With Stenosis Prevention Therapy.
The dominant location of electrical triggers for initiating atrial fibrillation (AF) originates from the muscle sleeves inside pulmonary veins (PVs). Currently, radiofrequency ablation (RFA) is performed outside of the PVs to isolate, rather than directly ablate these tissues, due to the risk of intraluminal PV stenosis. In 4 chronic canine experiments, we performed direct PV muscle sleeve RFA ± postablation drug-coated balloon (DCB) treatment with paclitaxel/everolimus. Of the 4 PVs, 2 PVs were ablated and treated with DCB, 1 PV was ablated without DCB treatment (positive control), and 1 PV was left as a negative control. Local electrograms were assessed in PVs for near-field signals and were targeted for ablation. After 12-14 weeks survival, PVs were interrogated for absence of near-field PV potentials, and each PV was assessed for stenosis. All canines survived the study period without cardiorespiratory complications, and remained ambulatory. In all canines, PVs that were ablated and treated with DCB remained without any significant intraluminal stenosis. In contrast, PVs that were ablated and not treated with DCB showed near or complete intraluminal stenosis. At terminal study, PV potentials remained undetectable. A blinded, histologic analysis demonstrated that ablated PVs without DCB treatment had extensive thrombus, fibrin, mineralization, and elastin disruption. Our chronic canine data suggest that direct PV tissue ablation without subsequent stenosis is feasible with the use of postablation DCBs.


Transformation of spatial sensitivity along the ascending auditory pathway.
Locations of sounds are computed in the central auditory pathway based primarily on differences in sound level and timing at the two ears. In rats, the results of that computation appear in the primary auditory cortex (A1) as exclusively contralateral hemifield spatial sensitivity, with strong responses to sounds contralateral to the recording site, sharp cutoffs across the midline, and weak, sound-level-tolerant responses to ipsilateral sounds. We surveyed the auditory pathway in anesthetized rats to identify the brain level(s) at which level-tolerant spatial sensitivity arises. Noise-burst stimuli were varied in horizontal sound location and in sound level. Neurons in the central nucleus of the inferior colliculus (ICc) displayed contralateral tuning at low sound levels, but tuning was degraded at successively higher sound levels. In contrast, neurons in the nucleus of the brachium of the inferior colliculus (BIN) showed sharp, level-tolerant spatial sensitivity. The ventral division of the medial geniculate body (MGBv) contained two discrete neural populations, one showing broad sensitivity like the ICc and one showing sharp sensitivity like A1. Dorsal, medial, and shell regions of the MGB showed fairly sharp spatial sensitivity, likely reflecting inputs from A1 and/or the BIN. The results demonstrate two parallel brainstem pathways for spatial hearing. The tectal pathway, in which sharp, level-tolerant spatial sensitivity arises between ICc and BIN, projects to the superior colliculus and could support reflexive orientation to sounds. The lemniscal pathway, in which such sensitivity arises between ICc and the MGBv, projects to the forebrain to support perception of sound location.


Immune status of mice tolerant of living cells. II. Continuous presence and nature of facilitation-enhancing antibodies in tolerant animals.
CBA mice were rendered highly tolerant to A/Jax cells by neonatal intravenous injections of (CBA x A)F(1) spleen cells. The high degree of tolerance was ascertained by the absence of circulating antibodies detected in the sera by the usual tests and by the perfect state of A skin grafts during all the experiments. Tolerant sera (sera from tolerant animals) were studied at three periods of tolerance: before skin test grafting, from 2 to 11 wk after grafting, and at time of sacrifice at almost 6 months of age. The tolerant sera were shown to have specific facilitation-enhancing properties promoting the take and growth of A/Jax sarcoma (SaI and /Sa 15091a grafted on normal CBA mice. These properties were present throughout the duration of the experiments, showing that they were not the result of a beginning interruption of tolerance. The tolerant sera, although lacking the usual serological properties (hemagglutination, hemolysis, cytotoxicity, passive cutaneous anaphylaxis) had, however, specific synergistic hemagglutinating properties (increasing the hemagglutinating titer of a reference immune serum). Antibodies giving direct specific hemagglutination could be extracted from spleens of 20% of highly tolerant mice. The tolerant sera were also found to contain more IgG1 and more IgA than normal sera while they contained normal quantities of the complement-fixing immunoglobulins IgG2 and IgM. Fractionation of tolerant sera on DEAE chromatography column confirmed the data concerning immunoglobulin classes and demonstrated direct specific serological activities undetected in unfractionated sera: a weak hemolysis in the most cationic fractions and a weak hemagglutination in the middle fractions. Synergistic hemagglutination, detected in unfractionated serum, was localized in fast anionic fractions containing high IgA concentration, along with facilitation-enhancing activity, thus confirming a link suggested previously between these three properties. The relation between immunological tolerance and facilitating antibodies was discussed in the light of the fact that antibodies, possibly of a particular class continuously present at low dose in the sera of highly tolerant animals, are able to transfer (at least partly) this state of tolerance provided a sensitive test system is utilized.


Blood Pressure Variability May Be a New Predictor for the Occurrence and Prognosis of Ischemic Stroke.
Despite declines in morbidity and mortality in recent years, ischemic stroke (IS) remains one of the leading causes of death and disability from cerebrovascular diseases. Addressing the controllable risk factors underpins the successful clinical management of IS. Hypertension is one of the most common treatable risk factors for IS and is associated with poor outcomes. Ambulatory blood pressure monitoring has revealed that patients with hypertension have a higher incidence of blood pressure variability (BPV) than those without hypertension. Meanwhile, increased BPV has been identified as a risk factor for IS. The risk of IS is higher and the prognosis after infarction is worse with higher BPV, no matter in the acute or subacute phase. BPV is multifactorial, with alterations reflecting individual physiological and pathological changes. This article reviews the current research advances in the relationship between BPV and IS, with an attempt to raise awareness of BPV among clinicians and IS patients, explore the increased BPV as a controllable risk factor for IS, and encourage hypertensive patients to control not only average blood pressure but also BPV and implement personalized blood pressure management.


Individualized management of hepatic diseases in hereditary hemorrhagic telangiectasia.
Liver involvement in patients with hereditary hemorrhagic telangiectasia (HHT) has not been fully characterized in China. The clinical manifestations, imaging studies, results of treatment in six patients and symptomatic liver involvement were analyzed. Patients included three women and three men with age from 35 to 62 years old. Two patients presented with shortness of breath, one patient with anemia and splenomegaly, and one with chronic gastrointestinal bleeding; the remaining two were asymptomatic. CT and CT angiography (CTA) showed arterioportal and arteriovenous shunting in liver. CTA showed at least one enlarged hepatic artery in all patients. One patient received ligation of the enlarged arteries with subsequent disappearance of symptoms at 56-month follow-up. The patient with gastrointestinal bleeding received interventional embolotherapy and resolved; interventional therapy to embolize the enlarged hepatic arteries was unsuccessful in another patient and the patient died of heart failure and liver dysfunction 38 months later. The patient with splenomegaly received a splenectomy and bandage of an enlarged hepatic artery. One of the two patients with no symptoms died of liver dysfunction 41 months after diagnosis. The other showed abnormal liver function and ascites, and traditional Chinese medicinal herb was used with no effect 21 months later. The symptoms disappeared after systemic medical treatment. Individualized and active therapy is advantageous and proper for patients with HHT.


Reference values for skin microanatomy: A systematic review and meta-analysis of ex vivo studies.
Few studies have characterized reference values of normal human skin microanatomy parameters. To quantify histologic measurements of epidermal thickness, melanocyte density, hair follicle density, and eccrine gland density as a function of age and anatomic site. We searched the PubMed, Embase, Web of Science, and Cochrane databases for articles published through May 25, 2017. Two reviewers independently screened 2016 articles; 327 relevant articles and 151 additional articles found via forward or reference citations underwent full-text review by 1 of 4 reviewers for relevance, data extraction, and critical appraisal. Weighted averages, meta-analysis, and meta-regression were used in statistical analysis. A total of 56 articles were included; when all anatomic locations were used, the overall estimates for epidermal thickness, melanocyte density, hair follicle density, and eccrine gland density were 99.75 μm (95% confidence interval [CI], 83.25-116.25), 955.05 cells/mm2 (95% CI. 880.89-1029.21), 1.40 hairs/mm2 (95% CI. 0.91-1.89), and 1.28 glands/mm2 (95% CI. 0.91-1.64), respectively. There was significant data heterogeneity across studies, possibly because of differences in histological techniques and absence of standardized microanatomy definitions. We established summary estimates for normal human skin microanatomy parameters.


Dietetics students' perceptions of classroom-based learning activities.
The Accreditation Council for Education in Nutrition and Dietetics requires that undergraduate dietetics courses utilize a variety of educational approaches to facilitate learning. The aim of this pilot study was to evaluate undergraduate nutrition students' perceptions of 16 classroom-based learning activities before and after taking an upper-level nutrition course. A survey was completed by students before and after taking an upper-level nutrition course, Methods in Nutrition Education, at a single university in the southwest region of the United States in fall 2016 and 2017. The survey included demographic questions and assessed students' perceptions of the helpfulness of 16 traditional and active classroom-based activities to learning. Perceptions were measured via Likert scale (1 = strongly disagree to 5 = strongly agree) at baseline and postcourse. Wilcoxon signed rank tests assessed changes in students' perceptions of learning activities from baseline to postcourse (significance = P < 0.05). Ninety-seven completed baseline surveys; 67 (69%) completed postcourse surveys. Observing professionals (median = 5), interviewing professionals (median = 5), and critical thinking (median = 4) were perceived as most helpful to learning postcourse. Students agreed critical thinking, integrating material from other courses, interviewing professionals, case studies, writing short reports and summaries, and group projects and activities were significantly more helpful postcourse compared to baseline (P < 0.05). Undergraduate nutrition students perceive a variety of classroom-based activities are helpful to learning including traditional (textbook readings, lectures) and active learning strategies (observation, practice). Instructors may consider implementing a variety of traditional and active learning strategies in upper-level nutrition and health-related courses to facilitate learning.


[Cryosurgery of focal lesions of the liver in patients requiring reoperation].
Cryosurgery ++ (cryoresection and cryodestruction) was carried out in 21 of a group of 43 patients (48.8%) during resection and +re-resection of the liver in repeated operations. One patient died. It was found that cryodestruction combined with radical and palliative resections of the liver in reoperations improved the late-term results of the operation significantly. With the use of cryotechniques, blood loss is reduced and intraoperative dissemination of the tumour cells and alveococcus and recurrences in the postoperative period are prevented.


Immune vulnerability of infants to tuberculosis.
One of the challenges faced by the infant immune system is learning to distinguish the myriad of foreign but nonthreatening antigens encountered from those expressed by true pathogens. This balance is reflected in the diminished production of proinflammatory cytokines by both innate and adaptive immune cells in the infant. A downside of this bias is that several factors critical for controlling Mycobacterium tuberculosis infection are significantly restricted in infants, including TNF, IL-1, and IL-12. Furthermore, infant T cells are inherently less capable of differentiating into IFN- γ -producing T cells. As a result, infected infants are 5-10 times more likely than adults to develop active tuberculosis (TB) and have higher rates of severe disseminated disease, including miliary TB and meningitis. Infant TB is a fundamentally different disease than TB in immune competent adults. Immunotherapeutics, therefore, should be specifically evaluated in infants before they are routinely employed to treat TB in this age group. Modalities aimed at reducing inflammation, which may be beneficial for adjunctive therapy of some forms of TB in older children and adults, may be of no benefit or even harmful in infants who manifest much less inflammatory disease.


Intestinal bypass in morbid obesity: long-term metabolic sequelae.
A review of 51 patients undergoing jejunoileal bypass for morbid obesity is presented. There were two postoperative deaths; the 49 remaining patients have been followed for up to five years. Weight loss has been acceptable, the complications for the most part being transient and easily controlled. We believe that jejunoileal bypass for morbid obesity is a useful treatment modality in carefully selected patients.


The heart of anaesthesiology: revitalising humanism in the age of technology.
Technological innovation has greatly aided modern medicine, and anaesthesiology in particular, but also contributes to dehumanising influences that promote physician burnout and dissatisfaction among patients. Here we advocate for a profound reaffirmation of humanistic principles-empathy, compassion, and communication-in perioperative medicine. We propose adaptable strategies to bolster humanism in practice, such as curricular offerings, simulation training, role modelling, and recognition. As perioperative technologies continue to evolve, the threat of depersonalisation in anaesthetic care looms, making commitments to humanism a crucial precondition for healing in the communities in which we work and live.


Parkin gene modifies the effect of RLS4 on the age at onset of restless legs syndrome (RLS).
A co-occurrence of restless legs syndrome (RLS) and Parkin mutations has been described. In South Tyrolean RLS patients, a novel RLS locus has been found (RLS4) and recurrent Parkin mutations have been reported. By a systematic screen we investigated the presence of founder Parkin mutations in South Tyrolean RLS patients with known carrier status at the RLS4 locus and assessed whether these mutations alone or in combination influence the RLS phenotype measured by three quantitative RLS traits (age at onset (AAO) and two severity measurements). The Parkin mutation alone showed no effect, whereas RLS4 had a significant effect on the AAO (P = 0.0096, decrease of AAO of 9.1 years), but did not influence severity. Carriers of both, a Parkin mutation and the RLS4 haplotype, showed an association with AAO (P = 0.0016), corresponding to an anticipation of RLS onset age of 16.9 years. However, there was no effect on the disease severity. Our results suggest that the occurrence of a heterozygous Parkin mutation works in tandem with the gene at the RLS4 locus to lower the AAO in RLS.


Correlation of peripheral white cell and bone marrow changes with folate levels in pregnancy and their clinical significance.
Two hundred and thirty-eight ;megaloblastic' pregnancies are reviewed. Bone-marrow aspiration was performed in every case. Serum folate estimations were strongly suggestive of folic acid deficiency in only 34% of cases. If, however, the red cell folate was determined as well, there was an almost complete agreement with the changes in the erythroblasts. It is suggested that hyper-segmentation of the neutrophils in association with a diminished red cell folate level is indicative of significant folic acid deficiency. This series possibly shows a slight but statistically insignificant increase of accidental haemorrhage. No effect on birth weight or incidence of stillbirth was demonstrable.


The association of modernization with dyslipidaemia and changes in lipid levels in the Polynesian population of Western Samoa.
Obesity and non-insulin-dependent diabetes mellitus (NIDDM) have increased in prevalence in Polynesian Western Samoans over the 13-year period 1978-1991, as the population undergoes an 'epidemiological transition'. We therefore investigated changes in the frequency of dyslipidaemia over the same period in adults aged 25-74 years, and examined factors associated with dyslipidaemia in cross-sectional and longitudinal data. Subjects were drawn from three geographically defined locations representing different degrees of modernization. The age-standardized prevalence of dyslipidaemia increased in each location between 1978 (n = 1197) and 1991 (n = 1748) with the prevalence of hypercholesterolaemia (> or = 5.5 mmol/l) increasing from 18% to 36% (P < 0.001), and that of hypertriglyceridaemia (> or = 2.0 mmol/l) increasing from 9% to 15% (P < 0.001) in the capital city, Apia. In 1991 the highest serum concentrations of total, high density lipoprotein (HDL) and calculated low density lipoprotein (LDL) cholesterol were found in Poutasi (intermediate level of modernization), and the highest triglyceride levels in urbanized Apia. Higher levels of body mass index (BMI), waist-hip ratio (WHR), glucose intolerance, fasting insulin concentration, physical inactivity, educational level, and occupational status were all associated with adverse lipid levels in univariate data. Obesity (BMI in women, WHR in men) and survey location were the most important correlates of abnormal lipid levels in logistic regression models. Fasting insulin was also independently associated with high triglyceride levels in men, while in women the increasing levels of fasting insulin were associated with adverse levels of total, LDL and HDL cholesterol, and triglycerides. In longitudinal data (n = 311), lower baseline levels of cholesterol and triglycerides were associated with greater increases in either parameter at follow-up. Elevated fasting insulin and female gender also predicted increasing cholesterol concentrations, and urban residence predicted an increase in triglyceride levels. Current levels of dyslipidaemia in Western Samoa are similar to those observed in developed Western populations, and are increasing rapidly. These findings, considered along with the high prevalence of other cardiovascular disease risk factors in Samoans, including smoking, obesity and NIDDM, suggest that cardiovascular disease will be a major health concern in the future.


Prefrontal deficits in a murine model overexpressing the down syndrome candidate gene dyrk1a.
The gene Dyrk1a is the mammalian ortholog of Drosophila minibrain. Dyrk1a localizes in the Down syndrome (DS) critical region of chromosome 21q22.2 and is a major candidate for the behavioral and neuronal abnormalities associated with DS. PFC malfunctions are a common denominator in several neuropsychiatric diseases, including DS, but the contribution of DYRK1A in PFC dysfunctions, in particular the synaptic basis for impairments of executive functions reported in DS patients, remains obscure. We quantified synaptic plasticity, biochemical synaptic markers, and dendritic morphology of deep layer pyramidal PFC neurons in adult mBACtgDyrk1a transgenic mice that overexpress Dyrk1a under the control of its own regulatory sequences. We found that overexpression of Dyrk1a largely increased the number of spines on oblique dendrites of pyramidal neurons, as evidenced by augmented spine density, higher PSD95 protein levels, and larger miniature EPSCs. The dendritic alterations were associated with anomalous NMDAR-mediated long-term potentiation and accompanied by a marked reduction in the pCaMKII/CaMKII ratio in mBACtgDyrk1a mice. Retrograde endocannabinoid-mediated long-term depression (eCB-LTD) was ablated in mBACtgDyrk1a mice. Administration of green tea extracts containing epigallocatechin 3-gallate, a potent DYRK1A inhibitor, to adult mBACtgDyrk1a mice normalized long-term potentiation and spine anomalies but not eCB-LTD. However, inhibition of the eCB deactivating enzyme monoacylglycerol lipase normalized eCB-LTD in mBACtgDyrk1a mice. These data shed light on previously undisclosed participation of DYRK1A in adult PFC dendritic structures and synaptic plasticity. Furthermore, they suggest its involvement in DS-related endophenotypes and identify new potential therapeutic strategies.


Predictors of young people's use of sexual and reproductive health services in Nigeria: a mixed-method approach.
Sexual and Reproductive health Services (SRHS) are essential for the prevention and control of SRH problems among young people and the achievement of sustainable development goal 3. These services may be available but certain factors interfere with their access and utilization by the young people. This study sought to determine factors that predict the utilization of SRHS among young people in Enugu State, Nigeria. The study adopted mixed-method research employing a cross-sectional research design. The population of the study comprised young people between the ages of 12 and 22 years. A multi-stage sampling procedure was used to select 1447 young people used for the study. A questionnaire, in-depth interview, and focus group discussion were used for data collection. Percentages, Chi-square, and logistic regression were used to analyse quantitative data, while qualitative data were thematically analysed using NVivo software. Socio-demographic factors of gender, age, education, income, and living status (p = < .05) were significant predictors of utilization of SRHS. Psycho-cultural and health system factors (p = < .05) were also significant predictors of utilization of SRHS. The study concluded that some socio-demographic factors (of gender, age, level of education, income, and living status), psycho-cultural, and health system factors can be used to predict young people's utilization of SRHS. These predictors could be addressed through home sex education, regular training of health care providers on youth-friendly services delivery, and policy reforms.


Neuronal early endosomes require EHD1 for L1/NgCAM trafficking.
In neurons, the endosomal system is essential for membrane receptor trafficking to dendrites and axons and thereby participates in various neuronal functions, such as neurite outgrowth and synaptic plasticity. A multitude of regulators coordinates trafficking through endosomes, but most of them have not been studied in detail in neurons. In non-neuronal cells, EHD1 (Eps15 homology-domain containing protein 1) functions in the recycling endosome and is required for endosome-to-plasma membrane transport of multiple cargos. In this study, we analyze the role of EHD1 in neurons. In particular, we investigate whether EHD1 is required for polarized trafficking of the dendritically targeted transferrin and the axonal adhesion molecule L1/NgCAM (neuron-glia cell adhesion molecule) and, if so, in what compartment it is required. We find that endosomal recycling of both L1/NgCAM and transferrin is impaired when EHD1 is downregulated. We show that EHD1 colocalizes with L1/NgCAM and transferrin mostly in EEA1 (early endosome antigen 1)-positive early endosomes and less extensively with recycling endosomes. Using live imaging, we observe that EHD1 is stably associated with endosomal membranes during their maturation into EEA1-positive compartments and often persists on them longer than EEA1. Finally, we show that downregulation of EHD1 causes a delay of L1/NgCAM in exiting EEA1-positive endosomes, resulting in impaired targeting of L1/NgCAM to the axonal membrane. We conclude that, in neurons, EHD1 functions in early endosomes rather than (or possibly in addition to) recycling endosomes. These findings point to the existence of neuronal adaptations of the endosomal system.


Phospholipase C signaling tonically represses basal atrial natriuretic factor secretion from the atria of the heart.
The cardiac hormone atrial natriuretic factor (ANF or ANP) plays significant, well-established roles in a large number of physiological and pathophysiological processes, including water and electrolyte balance, blood pressure regulation, and cardiovascular growth. Understanding the regulation of its production and secretion by atrial cardiomyocytes is incomplete. We have previously established a significant role of G(i/o) protein signaling in modulating ANF secretion as promoted by stretch of the atrial myocardium. In the present study, we investigated the role of G(q) protein signaling and its relationship to G(i/o) protein signaling using pharmacological manipulation of proximal effectors of G(αq) in an ex vivo model of spontaneously beating rat atria. Phospholipase C (PLC) and protein kinase C (PKC) inhibitors dramatically increased basal secretion of ANF. Furthermore, although atrial wall stretch is a potent stimulus for secretion, stretch unexpectedly reduced ANF secretion to basal levels under PLC and PKC inhibitory conditions. Inhibition of the inositol triphosphate receptor did not appear to affect basal secretion but dose-dependently blocked stretch-secretion coupling. The results obtained demonstrate that the PLC and PKC signaling cascades play important albeit unexpected roles in the regulation of basal and stimulated ANF secretion and suggest interplay between the G(q) and G(i/o) protein signaling pathways.


Detection of nandrolone, testosterone, and their esters in rat and human hair samples.
Nandrolone and testosterone are anabolic androgenic steroids occasionally abused by athletes. A sensitive, specific, and reproducible gas chromatography-mass spectrometry method for the quantitative determination of nandrolone, testosterone, and their esters in hair has been developed. The limits of quantitation of this method, based on 20 mg of hair, were 50 pg/mg for nandrolone and testosterone, 100 pg/mg for testosterone acetate, and 200 pg/mg for nandrolone-decanoate. Nandrolone-d3 and testosterone-d3 were used as internal standards. This method has been applied to the analysis of these compounds incorporated into rat and human hair. Male Long-Evans rats were given nandrolone decanoate 60 mg/kg intraperitoneally (i.p.) once daily for 10 days over a time period of 14 days. Two of the three rats contained nandrolone in the pigmented hair collected at day 21 at a concentration of 63 and 76 pg/mg, respectively. No drug was found in the corresponding nonpigmented hair. The rat hair samples that tested positive for nandrolone contained also nandrolone decanoate in concentrations of 0.9 and 1.2 ng/mg, respectively. In a separate experiment rats were given testosterone acetate 10 mg/kg i.p. once daily for five days. No testosterone or testosterone acetate was detected in the rat hair samples. Hair specimens were also obtained from four self-reported steroid users. The hair of two subjects were determined to be positive for testosterone in concentrations of 54 and 81 pg/mg. These data demonstrate that it is possible to detect the steroids nandrolone, testosterone, and nandrolone decanoate in hair after systemic administration.


Randomized Phase IIB Trial of Proton Beam Therapy Versus Intensity-Modulated Radiation Therapy for Locally Advanced Esophageal Cancer.
Whether dosimetric advantages of proton beam therapy (PBT) translate to improved clinical outcomes compared with intensity-modulated radiation therapy (IMRT) remains unclear. This randomized trial compared total toxicity burden (TTB) and progression-free survival (PFS) between these modalities for esophageal cancer. This phase IIB trial randomly assigned patients to PBT or IMRT (50.4 Gy), stratified for histology, resectability, induction chemotherapy, and stage. The prespecified coprimary end points were TTB and PFS. TTB, a composite score of 11 distinct adverse events (AEs), including common toxicities as well as postoperative complications (POCs) in operated patients, quantified the extent of AE severity experienced over the duration of 1 year following treatment. The trial was conducted using Bayesian group sequential design with three planned interim analyses at 33%, 50%, and 67% of expected accrual (adjusted for follow-up). This trial (commenced April 2012) was approved for closure and analysis upon activation of NRG-GI006 in March 2019, which occurred immediately prior to the planned 67% interim analysis. Altogether, 145 patients were randomly assigned (72 IMRT, 73 PBT), and 107 patients (61 IMRT, 46 PBT) were evaluable. Median follow-up was 44.1 months. Fifty-one patients (30 IMRT, 21 PBT) underwent esophagectomy; 80% of PBT was passive scattering. The posterior mean TTB was 2.3 times higher for IMRT (39.9; 95% highest posterior density interval, 26.2-54.9) than PBT (17.4; 10.5-25.0). The mean POC score was 7.6 times higher for IMRT (19.1; 7.3-32.3) versus PBT (2.5; 0.3-5.2). The posterior probability that mean TTB was lower for PBT compared with IMRT was 0.9989, which exceeded the trial's stopping boundary of 0.9942 at the 67% interim analysis. The 3-year PFS rate (50.8% v 51.2%) and 3-year overall survival rates (44.5% v 44.5%) were similar. For locally advanced esophageal cancer, PBT reduced the risk and severity of AEs compared with IMRT while maintaining similar PFS.


Identification and expression analysis of herpes B virus-encoded small RNAs.
Herpes B virus (BV) naturally infects macaque monkeys and is genetically similar to herpes simplex virus (HSV). Zoonotic infection of humans can cause encephalitis and if untreated has a fatality rate of ∼80%. The frequent use of macaques in biomedical research emphasizes the need to understand the molecular basis of BV pathogenesis with a view toward improving safety for those working with macaques. MicroRNAs (miRNAs) are small noncoding RNAs that regulate the expression of mRNAs bearing complementary target sequences and are employed by viruses to control viral and host gene expression. Using deep sequencing and validation by expression in transfected cells, we identified 12 novel BV-encoded miRNAs expressed in lytically infected cells and 4 in latently infected trigeminal ganglia (TG). Using quantitative reverse transcription-PCR (RT-qPCR), we found that most of the miRNAs exhibited a high level of abundance throughout infection. Further analyses showed that some miRNAs could be generated from multiple transcripts with different kinetic classes, possibly explaining detection throughout infection. Interestingly, miRNAs were detected at early times in the absence of viral gene expression and were present in purified virions. In TG, despite similar amounts of viral DNA per ganglion, it was notable that the relative amount of each miRNA varied between ganglia. The majority of the miRNAs are encoded by the regions that exhibit the most sequence differences between BV and HSV. Additionally, there is no sequence conservation between BV- and HSV-encoded miRNAs, which may be important for the differences in the human diseases caused by BV and HSV.


Intermittent feeding in a migratory omnivore: digestion and body composition of American black duck during autumn.
Birds fast intermittently during weather disturbances and migration. We tested responses of black duck to lost feeding days during autumn mass gain. Nine adult males were fed a pelleted diet (1.5% fat, 15.8% protein, and 18.3% neutral detergent fiber) and caged indoors during September and October (12 h light; 17 degrees -24 degrees C) to measure balances over 14 d when fed ad lib. each day and fasted intermittently for 2 d wk(-1) (short fast) or 4 d wk(-1) (long fast). Body mass (1,081 g), body water content, and metabolizable intakes of energy and protein were maintained as daily intakes of dry matter increased to 1.65 (short fast) and 2.35 (long fast) times the unfasted level. Intermittent feeding reduced metabolizability of dry matter, energy, protein, and acid detergent fiber. Concentrations of Mn provided similar estimates of metabolizability to direct measures in unfasted birds but underestimated measures of birds on long fasts. Fasting regimes continued outdoors for 9 wk when temperatures declined to -9 degrees C. Birds on short fasts were heavier (1,373 vs. 1,241 g) and fatter (159 vs. 58 g) than those on long fasts, while body water (894 g) and protein (316 g) were similar between groups after 5 wk. Birds on long fasts subsequently gained mass when fed daily, but those on short fasts lost mass when fed each day. Omnivorous waterfowl combine ingestive and digestive flexibility with plasticity of body lipid to contend with uncertain food availability.


A generalized distance function for preferential choices.
Many cognitive theories of judgement and decision making assume that choice options are evaluated relative to other available options. The extent to which the preference for one option is influenced by other available options will often depend on how similar the options are to each other, where similarity is assumed to be a decreasing function of the distance between options. We examine how the distance between preferential options that are described on multiple attributes can be determined. Previous distance functions do not take into account that attributes differ in their subjective importance, are limited to two attributes, or neglect the preferential relationship between the options. To measure the distance between preferential options it is necessary to take the subjective preferences of the decision maker into account. Accordingly, the multi-attribute space that defines the relationship between options can be stretched or shrunk relative to the attention or importance that a person gives to different attributes describing the options. Here, we propose a generalized distance function for preferential choices that takes subjective attribute importance into account and allows for individual differences according to such subjective preferences. Using a hands-on example, we illustrate the application of the function and compare it to previous distance measures. We conclude with a discussion of the suitability and limitations of the proposed distance function.


Immunization of dissociated spleen cell cultures from normal mice.
A culture system for cell suspensions from mouse spleens has been described. The system provides adequate conditions for in vitro immunization on initial exposure to heterologous erythrocytes. The in vitro response closely parallels that observed in vivo with respect to size, early kinetics, antigen dose, and the inhibitory effect of passive antibody. The response of cultured cells differs in two respects from that seen in vivo. There is an increase in the ability to discriminate between different varieties of homologous erythrocytes and the in vitro response does not appear to be limited by whatever mechanisms regulate the in vivo response.


Mutational analysis of multiple lung cancers: Discrimination between primary and metastatic lung cancers by genomic profile.
In cases of multiple lung cancers, individual tumors may represent either a primary lung cancer or both primary and metastatic lung cancers. Treatment selection varies depending on such features, and this discrimination is critically important in predicting prognosis. The present study was undertaken to determine the efficacy and validity of mutation analysis as a means of determining whether multiple lung cancers are primary or metastatic in nature. The study involved 12 patients who underwent surgery in our department for multiple lung cancers between July 2014 and March 2016. Tumor cells were collected from formalin-fixed paraffin-embedded tissues of the primary lesions by using laser capture microdissection, and targeted sequencing of 53 lung cancer-related genes was performed. In surgically treated patients with multiple lung cancers, the driver mutation profile differed among the individual tumors. Meanwhile, in a case of a solitary lung tumor that appeared after surgery for double primary lung cancers, gene mutation analysis using a bronchoscopic biopsy sample revealed a gene mutation profile consistent with the surgically resected specimen, thus demonstrating that the tumor in this case was metastatic. In cases of multiple lung cancers, the comparison of driver mutation profiles clarifies the clonal origin of the tumors and enables discrimination between primary and metastatic tumors.


Risk factors for isolation of fluconazole and echinocandin non-susceptible Candida species in critically ill patients.
Introduction. Resistance rates to azoles and echinocandins of Candida spp. increased over the last decade.Hypothesis/Gap Statement. Widespread use of antifungals could lead to development and dissemination of resistant Candida spp.Aim. To identify risk factors for isolation of Candida spp. non-susceptible to either fluconazole or echinocandins.Methodology. All patients hospitalized in the Intensive Care Unit (ICU) of the University General Hospital of Patras, Greece with Candida spp. isolated from clinical specimens during a ten-year period (2010-19) were included. Candida isolates were identified using Vitek-2 YST card. Consumption of antifungals was calculated.Results. During the study period, 253 isolates were included. C. non-albicans predominated (64.4 %) with C. parapsilosis being the most commonly isolated (42.3 %) followed by C. glabrata (nomenclatural change to Nakaseomyces glabrataa; 8.7 %) and C. tropicalis (11.9 %). Among all isolates, 45.8 and 28.5 % were non-susceptible and resistant to fluconazole, respectively. Concerning echinocandins, 8.7 % of isolates were non-susceptible to at least one echinocandin (anidulafungin or micafungin) and 3.1 % resistant. Multivariate analysis revealed that hospitalization during 2015-19, as compared to 2010-14, isolate being non-albicans or non-susceptible to at least one echinocandin was associated with isolation of fluconazole non-susceptible isolate. Administration of echinocandin, isolate being C. glabrata or C. tropicalis, or Candida spp. non-susceptible to fluconazole were independently associated with isolation of Candida spp. non-susceptible to at least one echinocandin. Fluconazole's administration decreased during the study period, whereas liposomal-amphotericin B's and echinoncandins' administration remained stable.Conclusion. Fluconazole's non-susceptibility increased during the study period, despite the decrease of its administration. Although echinocandins' administration remained stable, non-susceptibility among Candida spp. increased.


Examining antenatal health literacy in Ghana.
To explore Ghanaian pregnant women's understanding and recognition of danger signs in pregnancy, birth preparedness and complication readiness, and their understanding of newborn care. An exploratory, qualitative study design was used. Data were gathered through six focus group discussions with 68 pregnant women attending antenatal care at a busy urban hospital in Ghana. Qualitative and descriptive data were analyzed using SPSS version 21. Health literacy was used as the guiding framework to analyze the qualitative data. Data were analyzed in the content domains of (a) understanding and recognition of danger signs in pregnancy, (b) preparedness for childbirth, (c) understanding and recognition of danger signs in the newborn, and (d) appropriate and timely referral. Women in this study identified danger signs of pregnancy and in the newborn, but had difficulty interpreting and operationalizing information they received during antenatal care visits, indicating that health education did not translate to appropriate health behaviors. Cultural beliefs in alternative medicine, lack of understanding, and prior negative encounters with healthcare professionals may have led to underutilization of professional midwives for delivery and health services. Women in this study exhibited low health literacy by incorrectly interpreting and operationalizing health education received during antenatal care. With limited health literacy, pregnant women cannot fully comprehend the scope of services that a health system can provide for them and their families. Achieving the greatest impact with limited time in antenatal care is a challenge. Since antenatal care is widely available to pregnant women in Ghana, it is vital to reexamine the way antenatal education is delivered. Pregnant women must receive health information that is accurate and easy to understand in order to make informed health choices that will improve maternal and child health.


Morphologic, phenotypic, and genotypic similarities between primary tumors and corresponding 3D cell cultures grown in a repeatable system-preliminary results.
Three-dimensional (3D) cell cultures are the new frontier for reproducing the tumor micro-environment in vitro. The aims of the study were (1) to establish primary 3D cell cultures from canine spontaneous neoplasms and (2) to demonstrate the morphological, phenotypic and genotypic similarities between the primary canine neoplasms and the corresponding 3D cultures, through the expression of tumor differentiation markers. Seven primary tumors were collected, including 4 carcinomas and 3 soft tissue sarcomas. 3D cell cultures reproduced the morphological features of the primary tumors and showed an overlapping immunophenotype of the primary epithelial tumors. Immunohistochemistry demonstrated the growth of stromal cells and macrophages admixed with the neoplastic epithelial component, reproducing the tumor microenvironment. Mesenchymal 3D cultures reproduced the immunophenotype of the primary tumor completely in 2 out of 3 examined cases while a discordant expression was documented for a single marker in one case. No single nucleotide variants or small indel were detected in TP53 or MDM2 genes, both in primary tumors and in 3D cell cultures specimens. In one sample, MDM2 amplicons were preferentially increased in number compared to TP53 ones, indicating amplification of MDM2, detectable both in the primary tumor and in the corresponding cell culture specimen. Here we demonstrate a good cell morphology, phenotype and genetic profile overlap between primary tumors and the corresponding 3D cultures grown in a repeatable system.


Breast tissue classification using x-ray scattering measurements and multivariate data analysis.
This study utilized two radiation scatter interactions in order to differentiate malignant from non-malignant breast tissue. These two interactions were Compton scatter, used to measure the electron density of the tissues, and coherent scatter to obtain a measure of structure. Measurements of these parameters were made using a laboratory experimental set-up comprising an x-ray tube and HPGe detector. The breast tissue samples investigated comprise five different tissue classifications: adipose, malignancy, fibroadenoma, normal fibrous tissue and tissue that had undergone fibrocystic change. The coherent scatter spectra were analysed using a peak fitting routine, and a technique involving multivariate analysis was used to combine the peak fitted scatter profile spectra and the electron density values into a tissue classification model. The number of variables used in the model was refined by finding the sensitivity and specificity of each model and concentrating on differentiating between two tissues at a time. The best model that was formulated had a sensitivity of 54% and a specificity of 100%.


Does poor ovarian response to gonadotrophins predict early menopause? A retrospective cohort study with minimum of 10-year follow-up.
One in 10 women reach menopause before they are 45 years of age, and 1 in 100 before 40. In most cases, poor ovarian response to gonadotrophins is a result of poor ovarian reserve. An early menopause is associated with long-term health risks. Identifying women at risk may allow appropriate measures to be instigated early. Women aged <40 years treated in the Aberdeen Fertility Centre between 1998 and 2002 were identified. Those with poor response to an age appropriate dose of gonadotrophins (obtaining ≤3 eggs or had cycle cancelled) after exclusion of hypothalamic insufficiency, or whose cycle was cancelled due to poor response were age matched with good responders (6-15 eggs). In this retrospective cohort study, women who have had IVF at least 10 years ago (157 poor responders and 314 good responders) were sent a postal questionnaire to determine age at menopause. A total of 219 women (64 poor responders, 155 good responders) returned their questionnaires. Poor responders were more likely to have premature menopause (3% vs. 0%; p = .024). A higher proportion of poor responders experienced early menopause (11% vs. 3%; p = .044). Despite being the first study with a 10-year follow-up, this study is limited to one centre and has a small number of women reaching premature menopause. Poor response to gonadotrophins in the context of IVF treatment is a marker of reduced ovarian reserve and is associated with early menopause. Results of this study underline the need for larger studies with long-term follow-up.


A Study on the Use of Massive Open Online Courses in Otorhinolaryngology After Graduation.
The use of massive open online courses (MOOCs) in medical education and postgraduate education is becoming more and more widespread across the world. A survey study was carried out in Turkey to collect information and raise awareness about the use of MOOCs for educational and training purposes in the field of otorhinolaryngology after graduation. The prepared survey questionnaire was published on the online communication platform of the Turkish Otorhinolaryngology-Head and Neck Surgery Association, and volunteers were asked to fill-out the questionnaire by clicking the given link. A total of 210 volunteers participated in the survey study. Most of the volunteers (26.2%) were in the 36-40 age range. At the time of the study 53.8% of the volunteers were working as specialist doctors, 19.5% as residents, 10.5% as professors, 8.1% as associate professors, and 8.1% as assistant professors. Only 19 (9%) respondents ticked "yes" in response to the question "Are you familiar with the MOOC programs?" and only six (2.9%) of these respondents had attended a MOOC program. Evaluation of the correlation between academic title and awareness of MOOC programs showed that the instructors had a significantly higher level of MOOC awareness compared to the specialists and the residents (p=0.032). Results of the survey revealed that the use of MOOC programs in postgraduate otorhinolaryngology education was low and the volunteers were not well-informed about this practice.


A Rare Case of Nasal Lobular Capillary Hemangioma Centered at the Pyriform Aperture.
We present the rare case of a nasal lobular capillary hemangioma with bony erosion atypically localized to the left piriform aperture. This fast-growing lesion caused both physical and emotional distress due to recurrent epistaxis and prominent esthetic deformity evidenced by a large nasal growth. While the final diagnosis was benign, the clinical and radiological findings caused concern. This case serves as a reminder that prompt referral, work-up, and diagnosis are essential to optimize patient care.


Occupancy models for monitoring marine fish: a bayesian hierarchical approach to model imperfect detection with a novel gear combination.
Occupancy models using incidence data collected repeatedly at sites across the range of a population are increasingly employed to infer patterns and processes influencing population distribution and dynamics. While such work is common in terrestrial systems, fewer examples exist in marine applications. This disparity likely exists because the replicate samples required by these models to account for imperfect detection are often impractical to obtain when surveying aquatic organisms, particularly fishes. We employ simultaneous sampling using fish traps and novel underwater camera observations to generate the requisite replicate samples for occupancy models of red snapper, a reef fish species. Since the replicate samples are collected simultaneously by multiple sampling devices, many typical problems encountered when obtaining replicate observations are avoided. Our results suggest that augmenting traditional fish trap sampling with camera observations not only doubled the probability of detecting red snapper in reef habitats off the Southeast coast of the United States, but supplied the necessary observations to infer factors influencing population distribution and abundance while accounting for imperfect detection. We found that detection probabilities tended to be higher for camera traps than traditional fish traps. Furthermore, camera trap detections were influenced by the current direction and turbidity of the water, indicating that collecting data on these variables is important for future monitoring. These models indicate that the distribution and abundance of this species is more heavily influenced by latitude and depth than by micro-scale reef characteristics lending credence to previous characterizations of red snapper as a reef habitat generalist. This study demonstrates the utility of simultaneous sampling devices, including camera traps, in aquatic environments to inform occupancy models and account for imperfect detection when describing factors influencing fish population distribution and dynamics.


Detection of mucus glycoconjugates in human conjunctiva by using the lectin colloidal gold technique in TEM. I. A quantitative study in normal subjects.
We applied a specific cytochemical reaction to characterize the glycoconjugates produced by goblet and non-goblet epithelial cells of normal human conjunctiva. For this purpose we utilized the lectins, proteins of vegetal origin, which are extremely sensitive in binding glycosidic residues. In particular, we used WGA, PNA, SBA and ConA conjugated with colloidal gold as ultrastructural marker for Transmission Electron Microscopy. This technique allowed us also to perform a quantitative analysis, by counting colloidal gold particles present on mucus granules. In this way we analyzed the content both of goblet and non-goblet epithelial cells. In the former, WGA, PNA, SBA and ConA receptors, here reported in decreasing density, were present. In the latter WGA was always positive, SBA and PNA sometimes were negative, ConA was always negative. We speculate the different contribution to mucus production by these two sources may be important in evaluating tear film stability alterations occurring in those diseases in which non-goblet epithelial cell vesicles increase.


Development of a nomogram prognostic model for early Grade ≥ 3 infection in newly diagnosed multiple myeloma based on immunoparesis.
Infection, a significant cause of death in multiple myeloma (MM) patients, is a common complication and is closely associated with immunoparesis. There exists no clear definition of early infection, so early infection is defined in this paper as the occurrence within 3 months after diagnosis, considering the high incidence of infections within 3 months after diagnosis. This study established a new nomogram model based on immunoparesis to identify MM patients with high-risk early infection. A retrospective collection of 430 NDMM patients from June 2013 to June 2022 was conducted, and the patients were further divided into a training cohort and a validation cohort. In the training cohort, the least absolute shrinkage and selection operator (LASSO) was used to select the best variables that can be used to establish a new nomogram prediction model. Validation was performed in the validation and entire cohorts. After diagnosis, 67.7 % of the patients suffered from severe infection within 1 year, and 59.5 % experienced the first severe infection within 3 months. Variables associated with an increased risk of severe infection in the first 3 months included: BMPC, D-dimer, serum β2 microglobulin, immunoparesis, albumin, and eGFR. The nomogram based on the above six factors achieved a good C-index of 0.754, 0.73, and 0.731 in predicting early infection in the training cohort, validation cohort, and entire cohort, respectively. Finally, the time-dependent receiver operating characteristic (ROC) curve and decision curve analysis (DCA) of the nomogram showed that the model provided superior diagnostic capacity and clinical net benefit. In this study, we established a nomogram model to predict early grade ≥ 3 infection in NDMM patients. This model can assist clinicians in identifying NDMM patients with high-risk infections and improve their prognosis through early intervention.


Transrearrangements between antigen receptor genes in normal human lymphoid tissues and in ataxia telangiectasia.
The polymerase chain reaction (PCR) was used on DNA obtained from various normal lymphoid tissues to amplify chimeric TCR gene rearrangements involving J segments of the beta gene and V segments of the gamma or delta genes. As found previously for the transrearrangements between the gamma and delta genes, transrearrangements involving the beta gene were more abundant in DNA of the thymus than in DNA of the spleen, lymph node, bone marrow, or PBL. In addition, transrearrangements between Ig H chain V region segment and J segment of TCR delta chain were also found in DNA of normal thymus. Sequence analysis of the trans-rearrangement PCR products revealed structures closely resembling normal intragenic rearrangements, with N insertions and often D segments at the junctions between segments. The sequences analyzed suggest that transrearrangements arise through the action of normal lymphocyte recombinase, involve trans recognition of heptamer/nonamer recombination signals, and follow the 12 + 23 spacer rule. To test whether transrearrangements result from chromosomal rearrangements with breakpoints at the sites of Ag receptor genes, PCR was performed on the DNA of PBL from patients with ataxia telangiectasia, a disorder in which circulating lymphocytes often have numerous karyotypic abnormalities with breakpoints at the cytogenetic positions of these genes. Comparison of the results of PCR on this DNA and that of normal tissues demonstrated a substantially increased frequency for most types of transrearrangements investigated. These results support the interpretation that transrearrangement among TCR genes may occur by chromosomal rearrangement.


Attenuation of growth hormone production at the fetal stage is critical for dioxin-induced developmental disorder in rat offspring.
Although dioxins and related chemicals have been suspected to disrupt child development, their toxic mechanism remains poorly understood. Our previous studies in rat fetuses revealed that maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a highly-toxic dioxin, suppresses fetal synthesis of pituitary growth hormone (GH) that is essential for development. This study examined the hypothesis that attenuating GH expression in fetuses triggers developmental disorders. Treating pregnant rats with 1 μg/kg TCDD reduced the circulating level of GH and its downstream factor, insulin-like growth factor-1 (IGF-1), in the offspring only during the fetal and early neonatal stages. Although maternal TCDD exposure resulted in low body weight and length at babyhood and defects in the learning and memory ability at adulthood, GH supplementation in TCDD-exposed fetuses restored or tended to restore the defects including IGF-1 downregulation. Moreover, maternal TCDD exposure decreased the number of GH-positive cells during the fetal/neonatal stage. A microarray analysis showed that TCDD reduced the expression of death-associated protein-like 1 (DAPL1), a cell cycle-dependent proliferation regulator, in the fetal pituitary gland. In addition, TCDD treatment attenuated proliferating cells and cyclin mRNA expression in the fetal pituitary gland. Aryl hydrocarbon receptor (AHR)-knockout fetuses were insensitive to TCDD treatment, indicating that the TCDD-induced reduction in DAPL1 and GH mRNAs expression was due to AHR activation. Finally, DAPL1 knockdown suppressed GH and cyclin D2 expression in fetal pituitary cells. These results provide a novel evidence that dioxin suppresses GH-producing cell proliferation and GH synthesis due to partly targeting DAPL1, thereby impairing offspring development.


Investigation of atmospheric turbulence by narrow laser beams.
In order to determine the characteristic parameters of the atmospheric turbulence, the mutual dancing of two parallel narrow laser beams propagating through a turbulent atmosphere has been investigated theoretically and experimentally. The experiments have been carried out with a He-Ne laser on a 130-m path, one meter from the ground. The method allowed the determination of the variance of the refractive index fluctuations, as well as the values of scale of the turbulence in the vertical and horizontal planes.


Analysis of benzodiazepine withdrawal program managed by primary care nurses in Spain.
Benzodiazepine (BZD), the long-term treatment of which is harmful for cognitive function, is widely prescribed by General Practitioners in Spain. Based on studies performed in other countries we designed a nurse-led BZD withdrawal program adapted to Spanish Primary Care working conditions. A pseudo-experimental (before-after) study took place in two Primary Care Centres in Barcelona. From a sample of 1150 patients, 79 were identified. They were over 44 years old and had been daily users of BZD for a period exceeding six months. Out of the target group 51 patients agreed to participate. BZD dosage was reduced every 2-4 weeks by 25% of the initial dose with the optional support of Hydroxyzine or Valerian. The rating measurements were: reduction of BZD prescription, demographic variables, the Short-Form Health Survey (SF-12) to measure quality of life, the Medical Outcomes Study (MOS) Sleep Scale, and the Goldberg Depression and Anxiety Scale.By the end of the six-month intervention, 80.4% of the patients had discontinued BZD and 64% maintained abstinence at one year. An improvement in all parameters of the Goldberg scale (p <0.05) and in the mental component of SF-12 at 3.3 points (p = 0.024), as well as in most components of the MOS scale, was observed in the group that had discontinued BZD. No significant differences in these scales before and after the intervention were observed in the group that had not discontinued. At one year approximately 2/3 of the patients had ceased taking BZD. They showed an overall improvement in depression and anxiety scales, and in the mental component of the quality of life scale. There was no apparent reduction in the sleep quality indicators in most of the analysed components. Nurses in a Primary Care setting can successfully implement a BZD withdrawal program.


Morphologic and biochemical features affecting the antithrombotic properties of the inferior vena cava of rabbit pups and adult rabbits.
The incidence of venous thromboembolic disease is reduced in children compared with adults. Thromboprotective mechanisms, some of which have already been identified in plasma, must be present in children. Blood vessel walls have important antithrombotic properties that maintain blood fluidity. This is in part due to proteoglycan (PG)-related glycosaminoglycan (GAG) molecules within vessel walls. PGs are macromolecules with covalently attached GAG chains, either chondroitin, dermatan, heparan, or keratan sulfate. The influence of age on the concentration and anticoagulant activities of PGs and GAGs, within vein walls before puberty has not been previously investigated. We hypothesized that developmental differences in vein walls may contribute to the reduced risk of thrombosis in children. We used a rabbit model to examine morphologic and biochemical features of inferior venae cavae (IVCs). We assessed IVC wall morphology, PG distribution, GAG mass, and GAG antithrombin activity. Morphologically, there were only minor differences between pups and adult rabbits' IVCs. However, there was a significant increase in GAGs by mass in IVCs from pups compared with adult rabbits (p = 0.012). In addition the total antithrombin activity (p = 0.04), and especially that of heparan sulfate (p = 0.01) was significantly increased in pups compared with adult rabbits. These results demonstrate important differences in the antithrombotic properties of IVC walls in pups and adult rabbits. In summary, developmental differences in vein wall PG content and activity exist which may contribute to the reduced risk of venous thromboembolism in children. Further characterization of these differences is required.


Effect of the Xpert MTB/RIF on the detection of pulmonary tuberculosis cases and rifampicin resistance in Shanghai, China.
Xpert MTB/RIF (Xpert) is an automated molecular test recommended by World Health Organization (WHO) for diagnosis of tuberculosis (TB). This study evaluated the effect of Xpert implementation on the detection of pulmonary TB (PTB) and rifampicin-resistant TB (RR-TB) cases in Shanghai, China. Xpert was routinely implemented in 2018 for all presumptive PTB patients. All PTB patients above 15 years-old identified within the Provincial TB Control Program during the first half of each of 2017 and 2018, were enrolled to compare the difference in proportions of bacteriological confirmation, patients with drug susceptibility test (DST) results for rifampicin (ie, DST coverage) and RR-TB detection before and after Xpert's implementation. A total of 6047 PTB patients were included in the analysis with 1691 tested by Xpert in 2018. Percentages of bacteriological confirmation, DST coverage and RR-TB detection in 2017 and 2018 were 50% vs. 59%, 36% vs. 49% and 2% vs. 3%, respectively (all p-values < 0.05). Among 1103 PTB patients who completed sputum smear, culture and Xpert testing in 2018, Xpert detected an additional 121 (11%) PTB patients who were negative by smear and culture, but missed 248 (23%) smear and/or culture positive patients. Besides, it accounted for an increase of 9% in DST coverage and 1% in RR-TB detection. The median time from first visit to a TB hospital to RR-TB detection was 62 days (interquartile range -IQR 48-84.2) in 2017 vs. 9 days (IQR 2-45.7) in 2018 (p-value < 0.001). In the multivariate model, using Xpert was associated with decreased time to RR-TB detection (adjusted hazard ratio = 4.62, 95% confidence interval: 3.18-6.71). Integrating Xpert with smear, culture and culture-based DST in a routine setting significantly increased bacteriological confirmation, DST coverage and RR-TB detection with a dramatic reduction in the time to RR-TB diagnosis in Shanghai, China. Our findings can be useful for other regions that attempt to integrate Xpert into routine PTB and RR-TB case-finding cascade. Further study should focus on the identification and elimination of operational level challenges to fully utilize the benefit of rapid diagnosis by Xpert.


Prognostic variables in adult Wilms tumour.
To identify outcomes and prognostic variables that predict survival outcomes in adult Wilms tumour patients. We collected data on 128 patients with adult Wilms tumour treated between 1973 and 2006. Six cases from our 2 Canadian centres have not been previously reported. We collected data on the remaining 122 patients from published case reports or case series. Analyzed factors included age, sex, favourable or unfavourable histopathology, clinical stage (I, II, III or IV) and chemotherapy and radiotherapy received. The outcomes studied included overall survival (OS) and disease-specific survival (DSS). Univariate analysis with Kaplan-Meier actuarial methodology and multivariate analyses with Cox regression were used to determine outcomes and predictive clinical factors. The patients' mean age was 26 (range 15-73) years. After a mean follow-up of 54 (range 2-240) months, the OS and DSS of the entire cohort were both 68%. Favourable histopathology predicted superior OS and DSS (both p < 0.001). Higher clinical stage predicted inferior OS and DSS (both p < 0.001). Adult Wilms tumour has a poorer prognosis than pediatric Wilms tumour. In adults with Wilms tumour, more aggressive patient-and tumour-specific surveillance and adjunctive therapies than those advocated by pediatric National Wilms Tumor Study guidelines may be warranted, especially in patients with an unfavourable histopathology and higher clinical stage.


Effect of subinhibitory concentration of piperacillin/tazobactam on Pseudomonas aeruginosa.
Subinhibitory concentrations (sub-MICs) of antibiotics, although not able to kill bacteria, can modify their physico-chemical characteristics and the architecture of their outermost surface and may interfere with some bacterial functions. This study investigated the ability of sub-MIC piperacillin/tazobactam (P/T) to interfere with the bacterial virulence parameters of adhesiveness, cell-surface hydrophobicity, motility, biofilm formation and sensitivity to oxidative stress. Antimicrobial activity against five Pseudomonas aeruginosa clinical isolates, representative of clonal lineages of 96 strains of nosocomial origin, and six control strains (ATCC 27853, PAO1, AK1, MT1562, PT623, PAO1algC) was evaluated in vitro using the NCCLS microdilution method. The effects of sub-MIC on bacterial adhesion and biofilm formation were studied using a modified microtitre plate assay. The relative cell-surface hydrophobicity of P. aeruginosa strains was determined by measuring their ability to adhere to n-hexadecane. P. aeruginosa that had been exposed overnight to P/T and incubated with P/T in the plate were also screened for their ability to swim using flagella and to twitch and for their sensitivity to oxidative stress. The results obtained showed that the impact of sub-MIC P/T on bacterial characteristics was different for the various strains of P. aeruginosa. There was a change in bacterial morphology and hydrophobicity that could explain a significant decrease in adhesion values in all clinical isolates and controls tested, a decrease in biofilm formation, a significant increase in sensitivity to oxidative stress, a significant decrease in flagellum-mediated swimming and a decrease in type IV fimbriae-mediated twitching. The results obtained indicate that sub-MIC P/T interferes with the pathogenic potential of P. aeruginosa.


Cancer in pregnancy: gaps, challenges and solutions.
Cancer is the second leading cause of death during the reproductive years complicating between 0.02% and 0.1% of pregnancies. This incidence is expected to rise with the increase in age of childbearing. The relatively rare occurrence of pregnancy-associated cancer precludes conducting large, prospective studies to examine diagnostic, management and outcome issues. This article reviews the available data regarding the different aspects of the diagnosis and treatment of cancer during pregnancy as well as the effect of pregnancy on cancer prognosis. In pregnant patients diagnosed with cancer during the first trimester, treatment with multi-drug anti-cancer chemotherapy or radiotherapy (with fetal exposure >0.1-0.2 Gy) is associated with an increased risk of congenital malformations and therefore should follow a strong recommendation for pregnancy termination. The risk for malformation diminishes as pregnancy advances and when cancer is diagnosed during the second or third trimesters there is usually no clear indication for abortion. Treatment postponement, until achieving fetal maturity, while closely monitoring tumor growth may be considered in selected cases. According to the available experience it seems that non-obstetrical surgery may be performed during pregnancy without an increased risk for adverse outcomes. In most types of cancer, pregnancy has no effect on maternal prognosis when compared to non-pregnant patients matched by age, cancer stage and treatment.


Accurate Detection of Dysmorphic Nuclei Using Dynamic Programming and Supervised Classification.
A vast array of pathologies is typified by the presence of nuclei with an abnormal morphology. Dysmorphic nuclear phenotypes feature dramatic size changes or foldings, but also entail much subtler deviations such as nuclear protrusions called blebs. Due to their unpredictable size, shape and intensity, dysmorphic nuclei are often not accurately detected in standard image analysis routines. To enable accurate detection of dysmorphic nuclei in confocal and widefield fluorescence microscopy images, we have developed an automated segmentation algorithm, called Blebbed Nuclei Detector (BleND), which relies on two-pass thresholding for initial nuclear contour detection, and an optimal path finding algorithm, based on dynamic programming, for refining these contours. Using a robust error metric, we show that our method matches manual segmentation in terms of precision and outperforms state-of-the-art nuclear segmentation methods. Its high performance allowed for building and integrating a robust classifier that recognizes dysmorphic nuclei with an accuracy above 95%. The combined segmentation-classification routine is bound to facilitate nucleus-based diagnostics and enable real-time recognition of dysmorphic nuclei in intelligent microscopy workflows.


Pregnancy outcome following in utero exposure to lithium: a prospective, comparative, observational study.
The authors conducted a prospective, comparative observational study to evaluate the risk of major anomalies following exposure to lithium during pregnancy. A total of 183 lithium-exposed pregnancies of women who contacted the Israeli Teratology Information Service were followed up (90.2% in the first trimester) and compared with 72 disease-matched and 748 nonteratogenic-exposed pregnancies. There were significantly more miscarriages (adjusted odds ratio=1.94, 95% CI=1.08-3.48) and elective terminations of pregnancy (17/183 [9.3%] compared with 15/748 [2.0%]) in the lithium-exposed group compared with the nonteratogenic exposure group. The rate of major congenital anomalies after exclusion of genetic or cytogenetic anomalies was not significantly different between the three groups (lithium-exposed in the first trimester: 8/123 [6.5%]; bipolar: 2/61 [3.3%]; nonteratogenic: 19/711 [2.7%]). Cardiovascular anomalies occurred more frequently in the lithium group exposed during the first trimester when compared with the nonteratogenic exposure group (5/123 [4.1%] compared with 4/711 [0.6%]) but not after excluding anomalies that spontaneously resolved (3/123 [2.4%] compared with 2/711 [0.3%]). Ebstein's anomaly was diagnosed in one lithium-exposed fetus and in two retrospective lithium cases that were not included because contact with the information service was made after the prenatal diagnosis by ultrasound. The rate of noncardiovascular anomalies was not significantly different between the groups. The rate of preterm deliveries was higher in the lithium group compared with the nonteratogenic exposure group (18/131 [13.7%] compared with 41/683 [6.0%]). Lithium treatment in pregnancy is associated with a higher rate of cardiovascular anomalies. Women who are treated with lithium during organogenesis should undergo fetal echocardiography and level-2 ultrasound.


Equine atypical myopathy in the UK: Epidemiological characteristics of cases reported from 2011 to 2015 and factors associated with survival.
Equine atypical myopathy (AM) is a toxic rhabdomyolysis associated with ingestion of hypoglycin A, derived typically in Europe, from Acer pseudoplatanus tree. Despite the wide distribution of this tree species in the UK, the number of cases reported annually varies, and there has been an apparent increase in prevalence in recent years. Although AM was first recognised in the UK, epidemiological studies have never been conducted focused solely on this country. To describe the spatiotemporal distribution, presentation, treatment and outcome of AM cases reported in the UK. Retrospective case series. British AM cases reported to the atypical myopathy alert website, between 2011 and 2015 were included (n = 224). Data were obtained via standardised epidemiological questionnaires from owners and veterinarians. Factors associated with survival were assessed using logistic regression. Most cases reported were from England (87.9%). Survival was 38.6% (n = 73/189). Clinical factors associated with reduced odds of survival included, hypothermia (odds ratio [OR] 0.18; 95% confidence interval [CI] 0.06-0.57; P = 0.01), bladder distension (OR 0.11; CI 0.02-0.59; P = 0.01), tachycardia (OR 0.97; CI 0.94-0.99; P = 0.04) and serum creatine kinase activity >100,000 IU/L (OR 0.17; CI 0.04-0.68; P = 0.01) in the univariable analysis as well as recumbency. The latter was the only sign retained in multivariable analysis (OR = 0.19; CI 0.06-0.62; P = 0.006). Administration of vitamins during the disease was associated with survival (OR 3.75; CI 1.21-11.57; P = 0.02). Reporting cases to the Atypical Myopathy Alert Group is voluntary; therefore, under-reporting will result in underestimation of AM cases; furthermore, direct owner-reporting could have introduced misdiagnosis bias. Some areas of the UK reported AM cases more commonly. Clinical signs such as recumbency, rectal temperature, distended bladder and serum creatine kinase activity might be useful prognostic indicators though should be considered in the context of the clinical picture. Treatment with vitamins increases odds of survival.


A new perspective on the genetics of keratoconus: why have we not been more successful?
Twin studies and family studies suggest an important genetic basis for keratoconus (KC). Involvement and association of several genes with the disease has been reported. Additionally, genes associated with central corneal thickness (CCT) and corneal curvature (CC) via genome-wide association studies (GWAS), also potentially underlie KC. Although a long list of genes has been reported for KC, the evidence for a pathogenic role for most genes remains limited. Furthermore, if the involvement of the reported genes in KC development can be proven, they only account for a limited number of patients. VSX1, ZNF469, SOD1, and miR184 have been most frequently investigated, but only mutations in miR184 indisputably underlie corneal abnormalities. For the three other genes, analysis of the minor allele frequencies (MAF) in public databases argues against a pathogenic role for most reported variants. For the remainder of variants, functional evidence is needed to prove their contribution to the pathogenesis. Despite the large amount of studies, clear results remain rare. A possible explanation for the cumbersome gene-identification is that genetic defects underlying KC are located in regions that are understudied (such as non-coding regions) or that KC is not as monogenic (= one gene with large effect size) as initially considered. Since many of the applied research strategies can only identify large effect mutations, strategies to identify variants with smaller effect sizes might lead to more progress in KC research.


Pallidal deep brain stimulation in a 5-year-old child with dystonic storm: case report.
A 5-year-old child had a medical history of epilepsy and a newly presented mental retardation with a life-threatening dystonic storm. Neuroimagings showed bilateral calcification of the pallidum. Several treatment modalities were performed, but the symptoms showed no significant improvement. The patient was operated on in order to place a deep brain stimulation (DBS) targeting bilateral globus pallidum internus (GPi). The dystonia showed a remarkable improvement after surgery, with 81% reduction of dystonia severity after 15 months. To our best knowledge, this is the youngest patient mentioned in the literature to be treated with DBS, which was also life-saving in this case.


Effects of a brief mindfulness-based intervention program for stress management among medical students: the Mindful-Gym randomized controlled study.
Pursuing undergraduate medical training can be very stressful and academically challenging experience. A 5-week mindfulness-based stress management (MBSM/Mindful-Gym) program was developed to help medical students cope with stress. The aim of this study was to evaluate the effectiveness of the intervention in reducing stress among students in a medical school in Malaysia. Seventy-five medical students participated in the program. They were stratified according to years of studies and randomly allocated to intervention (N = 37) and control groups (N = 38). The following outcome variables were measured at pre- and post-intervention: mindfulness (with Mindful Awareness Attention Scale); perceived stress (with Perceived Stress Scale); mental distress (with General Health Questionnaire), and self-efficacy (with General Self-efficacy Scale). Hierarchical multiple regressions were used to analyse the effect of group (intervention vs. control) on changes in the outcome variables. There were significant improvements at one week post-intervention in all outcome variables: mindfulness (β = 0.19, ΔR2 = 0.04, p = .040, f (2) = 0.05), perceived stress (β = -0.26, ΔR2 = 0.07, p = .009, f (2) = 0.10); mental distress (β = -0.28, ΔR2 = 0.10, p = .003, f (2) = 0.15); and self-efficacy (β = 0.30, ΔR2 = 0.09, p < .001, f (2) = 0.21). Six months after the intervention, those who had joined the program reported higher self-efficacy compared to those in the control group (β = 0.24, ΔR2 = 0.06, p = .020, f (2) = 0.08); but there was no difference in other outcome measures. More than 90% of the participants found the program applicable in helping patients and all reported that they would recommend it to others. This study indicates that the program is potentially an effective stress management program for medical students in Malaysia.


Residual ligand entropy in the binding of p-substituted benzenesulfonamide ligands to bovine carbonic anhydrase II.
In studies on the thermodynamics of ligand-protein interactions, it is often assumed that the configurational and conformational entropy of the ligand is zero in the bound state (i.e., the ligand is rigidly fixed in the binding pocket). However, there is little direct experimental evidence for this assumption, and in the case of binding of p-substituted benzenesulfonamide inhibitors to bovine carbonic anhydrase II (BCA II), the observed thermodynamic binding signature derived from isothermal titration calorimetry experiments leads indirectly to the conclusion that a considerable degree of residual entropy remains in the bound ligand. Specifically, the entropy of binding increases with glycine chain length n, and strong evidence exists that this thermodynamic signature is not driven by solvent reorganization. By use of heteronuclear (15)N NMR relaxation measurements in a series (n = 1-6) of (15)N-glycine-enriched ligands, we find that the observed thermodynamic binding signature cannot be explained by residual ligand dynamics in the bound state, but rather results from the indirect influence of ligand chain length on protein dynamics.


Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation.
Gas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to develop a fully automated system for quantifying serum organic acids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The pretreatment CLAM-2030 device was connected to an LC-MS/MS system for processing serum under optimized conditions, which included derivatizing serum organic acids using 3-Nitrophenylhydrazine. The derivatized organic acids were separated on a reverse-phase Sceptor HD-C column and detected using negative-ion electrospray ionization multiple reaction monitoring MS. The automated pretreatment-LC-MS/MS system processed serum in less than 1 h and analyzed 19 serum organic acids, which are used to detect organic acidemias. The system exhibited high quantitative sensitivity ranging from approximately 2 to 100 µM with a measurement reproducibility of 10.4% CV. Moreover, a proof-of-concept validation of the system was performed using sera from patients with propionic acidemia (n = 5), methylmalonic acidemia (n = 2), and 3-methylcrotonylglycinuria (n = 1). The levels of marker organic acids specific to each disease were significantly elevated in the sera of the patients compared to those in control samples. The automated pretreatment-LC-MS/MS system can be used as a rapid in-hospital system to measure organic acid levels in serum for the diagnosis of organic acidemias.


Transplantation of islet cells.
In an effort to improve the viability of canine pancreatic islet cell allografts after cryopreservation, Cy A was used for graft pretreatment and as a general immunosuppressant on graft recipients who had previously undergone pancreatectomy. Using this combination in conjunction with cryopreservation, islet cell allografts exhibited good viability and long term function after freezing to minus 196 degrees C. These results might encourage the further use of this technique for transplantation.


Rationale and Methodology of Reprogramming for Generation of Induced Pluripotent Stem Cells and Induced Neural Progenitor Cells.
Great progress has been made regarding the capabilities to modify somatic cell fate ever since the technology for generation of induced pluripotent stem cells (iPSCs) was discovered in 2006. Later, induced neural progenitor cells (iNPCs) were generated from mouse and human cells, bypassing some of the concerns and risks of using iPSCs in neuroscience applications. To overcome the limitation of viral vector induced reprogramming, bioactive small molecules (SM) have been explored to enhance the efficiency of reprogramming or even replace transcription factors (TFs), making the reprogrammed cells more amenable to clinical application. The chemical induced reprogramming process is a simple process from a technical perspective, but the choice of SM at each step is vital during the procedure. The mechanisms underlying cell transdifferentiation are still poorly understood, although, several experimental data and insights have indicated the rationale of cell reprogramming. The process begins with the forced expression of specific TFs or activation/inhibition of cell signaling pathways by bioactive chemicals in defined culture condition, which initiates the further reactivation of endogenous gene program and an optimal stoichiometric expression of the endogenous pluri- or multi-potency genes, and finally leads to the birth of reprogrammed cells such as iPSCs and iNPCs. In this review, we first outline the rationale and discuss the methodology of iPSCs and iNPCs in a stepwise manner; and then we also discuss the chemical-based reprogramming of iPSCs and iNPCs.


Graded interference with FGF signalling reveals its dorsoventral asymmetry at the mid-hindbrain boundary.
Signalling by fibroblast growth factors (FGFs) at the mid-hindbrain boundary (MHB) is of central importance for anteroposterior neural patterning from the isthmic organiser. Graded suppression of FGF signalling by increasing amounts of a dominant negative FGF receptor provides evidence that in addition to anteroposterior patterning, FGF signalling is also involved in patterning along the dorsoventral axis at the MHB. FGF signalling at the MHB is required for the activation of the HH target gene spalt at the MHB. Our results indicate that FGF signalling mediates the competence of the MHB to activate spalt in response to SHH. This interdependence of the two signalling pathways is also found in the outbudding optic vesicle where HH requires functional FGF signalling to activate spalt in the proximal eye region.


Activation of transcription factors NF-kappaB and NF-IL-6 by human immunodeficiency virus type 1 protein R (Vpr) induces interleukin-8 expression.
Human immunodeficiency virus (HIV)-positive individuals express elevated levels of interleukin-8 (IL-8), which is believed to be responsible for some of the clinical manifestations occurring during AIDS. We report here that virion-derived HIV type 1 (HIV-1) protein R (Vpr) increased IL-8 expression in primary T cells and macrophages, as well as in the T-cell line Jurkat, the monocytic cell line U937, and the epithelial cell line A549. Vpr appeared to increase IL-8 expression and IL-8 promoter activity by activating transcription factors NF-kappaB and NF-IL-6. Elevated Vpr was also shown to increase transcription of the NF-kappaB and NF-IL-6 enhancer-containing viral promoters for HIV, cytomegalovirus, and simian virus 40, as well as increase the expression of IL-6 and IL-10 in primary macrophages and in A549 cells, tumor necrosis factor alpha expression in primary T cells, and IL-6 and gamma interferon expression in U937 cells. These results suggest a new role for Vpr in the pathogenesis of HIV infection, namely, the activation of transcription factors NF-IL-6 and NF-kappaB.


Cardiac medications and their association with cardiovascular events in incident dialysis patients: cause or effect?
There are no large randomized, controlled trials evaluating the efficacy of common cardioprotective medications on cardiovascular outcomes in the dialysis population. We aimed to determine the association between cardioprotective medications and a composite end point of cardiovascular events or death in an incident dialysis cohort. Medicare claims data were utilized to determine outcomes in participants of the Dialysis Morbidity and Mortality Wave 2 cohort. Information was gathered at baseline regarding demographics, comorbidities, medication use, and lab data. Cox proportional hazard models were developed to determine the association between cardioprotective medication use at baseline and the subsequent development of a composite of cardiovascular events or death. Two thousand two hundred thirty-four of 3044 individuals experienced a cardiovascular event or death. Use of the following medications at baseline was associated with an increased event rate: nitrates, antiplatelet agents, and warfarin. There was no association between the use of angiotensin-converting enzyme inhibitors, beta-blockers, or 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors and a composite cardiovascular outcome or death. Only use of a calcium channel blocker at baseline was associated with a reduced risk of events. This beneficial association did not persist when death was excluded as an outcome. Many of the associations observed varied substantially depending on the outcome utilized. Use of cohort data to determine an association between medication use and outcome is difficult because the indication for the medication confounds the observed association. Before widespread practice changes, rigorous clinical trials of common cardioprotective medications on cardiovascular morbidity and mortality in dialysis should be performed.


Implications of secondary aortic intervention after thoracic endovascular aortic repair for acute and chronic type B dissection.
Thoracic endovascular aortic repair (TEVAR) has become a mainstay of therapy for acute and chronic type B aortic dissection (TBAD). Dynamic aortic morphologic changes, untreated dissected aorta, and persistent false lumen perfusion have significant consequences for reintervention after TEVAR for TBAD. However, few reports contrast differences in secondary aortic intervention (SAI) after TEVAR for TBAD or describe their influence on mortality. This analysis examined incidence, timing, and types of SAI after TEVAR for acute and chronic TBAD and determined their impact on survival. All TEVAR procedures for acute and chronic TBAD (2005-2016) were retrospectively reviewed. Patients with staged (<30 days) or concomitant ascending aortic arch repair or replacement were excluded. Acuity was defined by symptom onset (0-30 days, acute; >30 days, chronic). SAI procedures were grouped into open (intended treatment zone or remote aortic site), major endovascular (TEVAR extension or endograft implanted at noncontiguous site), and minor endovascular (side branch or false lumen embolization) categories. Kaplan-Meier methodology was used to estimate freedom from SAI and survival. Cox proportional hazards were used to identify SAI predictors. TEVAR for TBAD was performed in 258 patients (acute, 49% [n = 128]; chronic, 51% [n = 130]). Mean follow-up was 17 ± 22 months with an overall SAI rate of 27% (n = 70; acute, 22% [28]; chronic, 32% [42]; odds ratio, 1.7; 95% confidence interval, 0.9-2.9; P = .07]. Median time to SAI was significantly less after acute than after chronic dissection (0.7 [0-12] vs 7 [0-91] months; P < .001); however, freedom from SAI was not different (1-year: acute, 67% ± 4%, vs chronic, 68% ± 5%; 3-year: acute, 65% ± 7%, vs chronic, 52% ± 8%; P = .7). Types of SAI were similar (acute vs chronic: open, 61% vs 55% [P = .6]; major endovascular, 36% vs 38% [P = .8]; minor endovascular, 21% vs 21% [P = 1]). The open conversion rate (either partial or total endograft explantation: acute, 10% [13/128]; chronic, 15% [20/130]; P = .2) and incidence of retrograde dissection (acute, 6% [7/128]; chronic, 4% [5/130]; P = .5) were similar. There was no difference in survival for SAI patients (5-year: acute + SAI, 55% ± 9%, vs acute without SAI, 67% ± 8% [P = .3]; 5-year: chronic + SAI, 72% ± 6%, vs chronic without SAI, 72% ± 7% [P = .7]). Factors associated with SAI included younger age, acute dissection with larger maximal aortic diameter at presentation, Marfan syndrome, and use of arch vessel adjunctive procedures with the index TEVAR. Indication for the index TEVAR (aneurysm, malperfusion, rupture, and pain or hypertension) or remote preoperative history of proximal arch procedure was not predictive of SAI. SAI after TEVAR for TBAD is common. Acute TBAD has a higher proportion of early SAI; however, chronic TBAD appears to have ongoing risk of remediation after the first postoperative year. SAI types are similar between groups, and the occurrence of aorta-related reintervention does not affect survival. Patients' features and anatomy predict need for SAI. These data should be taken into consideration for selection of patients, device design, and surveillance strategies after TEVAR for TBAD.


Tissue distribution, turnover, and glycosylation of the long and short growth hormone receptor isoforms in rat tissues.
Two isoforms of the GH receptor, the full-length receptor (GHRL) and a short isoform (GHRS) that lacks the transmembrane and intracellular domains of GHRL, have been analyzed in rat tissue extracts by Western blotting and immunoprecipitation. Although quantitative estimates of GHRS and GHRL based on coprecipitation of [125I]GH indicated similar amounts of both isoforms in tissue extracts, the 110 kDa band corresponding to GHRL was generally not detected on Western blots without enrichment by immunoprecipitation. Two bands with electrophoretic mobilities corresponding to 38 and 42 kDa were present in extracts prepared from liver, muscle, and adipocytes. Western blots of the GH binding protein in rat serum also revealed two bands, but these had electrophoretic mobilities corresponding to 44 and 52 kDa. After digestion by endoglycosidase F, a single band with an electrophoretic mobility corresponding to 31 kDa was detected in samples from adipocytes, liver or serum, indicating that GHRS retained in tissues is glycosylated less extensively than that in rat serum. Digestion with neuraminidase indicated that the smaller glycoproteins in tissue extracts lack sialic acid residues that are present in serum samples. Furthermore, endoglycosidase H degraded GHRS in liver extracts to a 31 kDa band but did not degrade serum samples, suggesting that tissues retain a high mannose form of GHRS. The abundance of GHRS or GHRL in tissues from male, virgin female, and pregnant rats was estimated from the amount of 125I-GH that was bound to each isoform after immunoprecipitation. Liver contained more than 10 times as much GHRS per gram of tissue as fat or muscle. In liver, muscle, and fat, the amount of GHRS exceeded that of GHRL, sometimes by as much as 6-fold. GHBP levels in serum of females exceeded those in males, and rose even higher in pregnant females. The abundance of GHRS in all tissue extracts paralleled serum levels. In muscle and fat, the levels of GHRL did not differ in male, female and pregnant rats, whereas in liver, the pattern was similar to the GHRS pattern. In all tissues, pools of GHRS exceeded those of GHRL by a factor that grew larger as tissue and serum levels increased. The half life of GHBP in serum was estimated to be 2.4 h in rats treated with cycloheximide, whereas that of GHRS was 20 min in liver and 8.5 h in fat. These results suggest that GHRS is synthesized in liver 8 times faster than it is released into serum, whereas synthesis in fat is less than 30% of the rate at which it is released into serum by all tissues. Therefore, liver appears to be the major source of GHBP in serum. Although secretion into the circulatory system accounts for little or perhaps none of its turnover in some tissues, GHRS pools in tissues do appear to be regulated, suggesting that GHRS may function primarily in the cells in which it is synthesized.


Clinically relevant concentrations of valproic acid modulate melatonin MT(1) receptor, HDAC and MeCP2 mRNA expression in C6 glioma cells.
C6 glioma cells were treated with clinically relevant concentrations of valproic acid (0.5 or 1.0 mM) for 1-7 days and RT-PCR used to examine expression of the melatonin MT(1) receptor and selected epigenetic modulators. Valproic acid caused significant time-dependent changes in the mRNA expression of the melatonin MT(1) receptor, histone deacetylase (HDAC) 1, 2 and 3, and methyl CpG binding protein 2 (MeCP2). A structurally distinct HDAC inhibitor, trichostatin A, also caused a significant concentration-dependent induction of melatonin MT(1) receptor mRNA expression, suggesting involvement of an epigenetic mechanism. The ability of clinical concentrations of valproic acid to significantly alter melatonin MT(1) receptor expression, suggests a role for this receptor in the diverse neuropharmacological and oncostatic effects of this agent.