Functional anatomy and muscle moment arms of the thoracic limb of an elite sprinting athlete: the racing greyhound (Canis familiaris).
We provide quantitative muscle-tendon architecture and geometry data for the racing greyhound thoracic limb. Muscle mass, belly length, fascicle lengths, pennation angles and moment arms were measured, as were tendon masses and lengths. Maximum isometric force and maximum power were estimated for muscles, and maximum stress and strain were estimated for tendons. Results are compared with other fast quadrupedal runners, and to previously published data in mixed-breed dogs. The implications of the functional adaptations of the greyhound thoracic limb for sprinting performance are discussed. The thoracic limb was found to benefit from a similar proportion of locomotor muscle mass to the pelvic limb, suggesting that it may be used to some extent in propulsion, or alternatively that stabilisation is very important in this animal. Extrinsic muscles, especially latissimus dorsi and pectoralis profundus, were predicted to be powerful and important for generating net positive work during accelerations. Proximal biarticular muscles show specialisation toward preventing collapse of the shoulder and elbow joints to enable strut-like limb function, or some form of dynamic control. Distal muscles did not appear specialised for elastic energy storage, a functional difference to pelvic limb muscles, and the equivalents in horse thoracic limbs. The greyhound thoracic limb appears to possess substantial differences from both that of more 'sub-maximal specialist' quadrupeds, and from the greyhound pelvic limb.


Cytochrome P450 and P-Glycoprotein-Mediated Interactions Involving African Herbs Indicated for Common Noncommunicable Diseases.
Herbal remedies are regularly used to complement conventional therapies in the treatment of various illnesses in Africa. This may be because they are relatively cheap and easily accessible and are believed by many to be safe, cause fewer side effects, and are less likely to cause dependency. On the contrary, many herbs have been shown to alter the pharmacokinetics of coadministered allopathic medicines and can either synergize or antagonize therapeutic effects as well as altering the toxicity profiles of these drugs. Current disease burden data point towards epidemiological transitions characterised by increasing urbanization and changing lifestyles, risk factors for chronic diseases like hypertension, diabetes, and cancer which often present as multimorbidities. As a result, we highlight African herb-drug interactions (HDIs) modulated via cytochrome P450 enzyme family (CYP) and P-glycoprotein (P-gp) and the consequences thereof in relation to antihypertensive, antidiabetic, and anticancer drugs. CYPs are enzymes which account for to up to 70% of drug metabolism while P-gp is an efflux pump that extrudes drug substrates out of cells. Consequently, regulation of the relative activity of both CYP and P-gp by African herbs influences the effective drug concentration at the site of action and modifies therapeutic outcomes.


Long-term alpha-tocopherol supplementation is associated with lower serum vascular endothelial growth factor levels.
We previously reported that daily supplementation with alpha-tocopherol reduced prostate cancer risk in a large, randomized trial, the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. One potential mechanism explaining this is that alpha-tocopherol inhibited tumor angiogenesis, an effect demonstrated in animal models. We evaluated whether long-term supplementation with alpha-tocopherol modified serum vascular endothelial growth factor (VEGF) levels, a cytokine integrally involved in angiogenesis, in men who were not diagnosed with cancer and had baseline and follow-up blood available. One hundred of these men who received alpha-tocopherol (50 mg daily) were randomly selected and matched on age, study center and time between blood draws to 100 men who received placebo (median follow-up 3.7 years). VEGF levels were measured by enzyme-linked immunosorbent assay. The effect of alpha-tocopherol supplementation on serum VEGF was evaluated using a matched-paired t-test for differences in the change in VEGF over the intervention period between groups. There was an 11% reduction in VEGF levels in the alpha-tocopherol group as compared with a 10% increase in the placebo group (p=0.03). Our findings suggest that one of the mechanisms behind the inhibition of prostate carcinogenesis by alpha-tocopherol in the ATBC Study may have been through reduced VEGF concentrations and the suppression of tumor angiogenesis and therefore growth.


Phospholipase D1 is threonine-phosphorylated in human-airway epithelial cells stimulated by sphingosine-1-phosphate by a mechanism involving Src tyrosine kinase and protein kinase Cdelta.
The regulatory role of protein kinase C (PKC) delta isoform in the stimulation of phospholipase D (PLD) by sphingosine-1-phosphate (SPP) in a human-airway epithelial cell line (CFNPE9o(-)) was revealed by using antisense oligodeoxynucleotide to PKCdelta, in combination with the specific inhibitor rottlerin. Cell treatment with antisense oligodeoxynucleotide, but not with sense oligodeoxynucleotide, completely eliminated PKCdelta expression and resulted in the strong inhibition of SPP-stimulated phosphatidic acid formation. Indeed, among the PKCalpha, beta, delta, epsilon and zeta isoforms expressed in these cells, only PKCdelta was activated on cell stimulation with SPP, as indicated by translocation into the membrane fraction. Furthermore, pertussis toxin and genistein eliminated both PKCdelta translocation and PLD activation. In particular, a significant reduction in phosphatidylbutanol formation by SPP was observed in the presence of 4-amino-5-(4-methylphenyl)-7-(t-butyl) pyrazolo [3,4-d] pyrimidine (PP1), an inhibitor of Src tyrosine kinase. Furthermore, the activity of Src kinase was slightly increased by SPP and inhibited by PP1. However, the level of PKCdelta tyrosine phosphorylation was not increased in SPP-stimulated cells, suggesting that Src did not directly phosphorylate PKCdelta. Finally, the level of serine phosphorylation of PLD1 and PLD2 isoforms was not changed, whereas the PLD1 isoform alone was threonine-phosphorylated in SPP-treated cells. PLD1 threonine phosphorylation was strongly inhibited by rottlerin, by anti-PKCdelta oligodeoxynucleotide and by PP1. In conclusion, in CFNPE9o(-) cells, SPP interacts with a membrane receptor linked to a G(i) type of G-protein, leading to activation of PLD, probably the PLD1 isoform, by a signalling pathway involving Src and PKCdelta.


Associations between radius low-frequency axial ultrasound velocity and bone fragility in elderly men and women.
An exploratory study in elderly women and men from the Geneva Retirees Cohort indicates that low-frequency quantitative ultrasound measurement at the radius captures aBMD, bone size, and cortical tissue mineral density and might be used for screening purposes prior to DXA to evaluate fracture risk. INTRODUCTION: The contribution of distal radius bone mineral density (BMD) and cortical microstructure to fracture risk has recently been demonstrated. In this exploratory study, we investigated whether low-frequency quantitative ultrasound measurement at the distal radius may capture the peripheral determinants of bone fragility assessed with dual-energy X-ray absorptiometry (DXA) and high-resolution peripheral quantitative computed tomography (HR-pQCT). Low-frequency velocity (VLF) was measured at the radius using OsCare Sono®, a portable axial transmission ultrasonometer, in 271 community-dwelling postmenopausal women and men (age 71.5 ± 1.4 years) from the Geneva Retirees Cohort. Cortical (Ct) and trabecular (Tb) volumetric (v) BMD and microstructure at the distal radius were assessed by HR-pQCT, in addition to areal (a) BMD by DXA, at the same time point. VLF was highly correlated with aBMD at the distal third radius (r = 0.72, p < 0.001). For microstructure parameters, the highest correlation was observed with cortical area (r = 0.59, p < 0.001). VLF also captured bone geometry (total area) and cortical tissue mineral density independently of aBMD. In models adjusted for age and sex, VLF was significantly associated with prevalent low-trauma fractures [OR 95%CI for one SD decrease of VLF 1.50 (1.05, 2.14), p = 0.024], with discrimination performance comparable to femoral neck or distal radius aBMD. Measurement of VLF at the radius captures aBMD, bone size, and cortical tissue mineral density and might be used for screening purposes prior to DXA to evaluate fracture risk.


A bis-sulfonyl O,C,O aryl pincer ligand and its tin(II) complex: synthesis, structural studies, and DFT calculations.
The efficiency of the deprotonated aryl bis-sulfone [2,6-{(p-tolyl)SO2}2C6H3](-) as an O,C,O-coordinating pincer-type ligand was described. The bis-sulfone precursor was synthesized using a straightforward palladium-catalyzed cross-coupling reaction. As a result of directed ortho metalation (DoM) through sulfonyl groups, a selective lithiation of the aryl group was achieved and the corresponding carbanion was isolated and its structure determined by single-crystal X-ray diffraction analysis. A heteroleptic tin(II) complex has been prepared by a nucleophilic substitution reaction. Crystallographic analysis and DFT calculations indicate that the bis-sulfonyl moiety acts as a new O,C,O-coordinating pincer-type ligand with intramolecular S=O coordination to a tin(II) center. The cis form with the two nonbonded oxygen atoms of the sulfonyl groups on the same side is preferentially obtained.


Impact of cyanidin-3-glucoside on glycated LDL-induced NADPH oxidase activation, mitochondrial dysfunction and cell viability in cultured vascular endothelial cells.
Elevated levels of glycated low density lipoprotein (glyLDL) are frequently detected in diabetic patients. Previous studies demonstrated that glyLDL increased the production of reactive oxygen species (ROS), activated NADPH oxidase (NOX) and suppressed mitochondrial electron transport chain (mETC) enzyme activities in vascular endothelial cells (EC). The present study examined the effects of cyanidin-3-glucoside (C3G), a type of anthocyanin abundant in dark-skinned berries, on glyLDL-induced ROS production, NOX activation and mETC enzyme activity in porcine aortic EC (PAEC). Co-treatment of C3G prevented glyLDL-induced upregulation of NOX4 and intracellular superoxide production in EC. C3G normalized glyLDL-induced inhibition on the enzyme activities of mETC Complex I and III, as well as the abundances of NADH dehydrogenase 1 in Complex I and cytochrome b in Complex III in EC. Blocking antibody for the receptor of advanced glycation end products (RAGE) prevented glyLDL-induced changes in NOX and mETC enzymes. Combination of C3G and RAGE antibody did not significantly enhance glyLDL-induced inhibition of NOX or mETC enzymes. C3G reduced glyLDL-induced RAGE expression with the presence of RAGE antibody. C3G prevented prolonged incubation with the glyLDL-induced decrease in cell viability and the imbalance between key regulators for cell viability (cleaved caspase 3 and B cell Lyphoma-2) in EC. The findings suggest that RAGE plays an important role in glyLDL-induced oxidative stress in vascular EC. C3G may prevent glyLDL-induced NOX activation, the impairment of mETC enzymes and cell viability in cultured vascular EC.


Chronic inflammation therapy combined with low dose cytokines in the treatment of chronic ulcers of lower limbs.
Prevalence of chronic venous leg ulcers has increased from 55% to 75% in 10 years. We treat chronic vascular lesions according to a cyclic multiphase method that follows the instrumental and differential etiologic diagnosis of ulcers of the lower limbs. The AIMED (Anti Inflammatory Regenerative Medicine) operating model follows the diagnostic phase and is structured in 5 interconnected phases. Mare evaluation, pH control, biofilm removal and prevention and infections are approached according to the International Guidelines. Low-dose cytokine therapy to contrast the inflammatory process are administrated to the patients. Chronic skin lesions of these patients, treated according to the AIMED protocol, have been evaluated twice a week. The clinical evaluation parameters evaluated by us were: reduction of pain at 15 and 30 days, reduction of 50% of the injury at 30 days, characteristics of perilesional skin (edema, erythema) and global profile of the patient. The so-called Low Dose Cytokines therapy is changing the previous therapeutic algorithms thanks to a safe and effective systemic cytokines-receptors interactive physiological process.


Importing MAGE-ML format microarray data into BioConductor.
The microarray gene expression markup language (MAGE-ML) is a widely used XML (eXtensible Markup Language) standard for describing and exchanging information about microarray experiments. It can describe microarray designs, microarray experiment designs, gene expression data and data analysis results. We describe RMAGEML, a new Bioconductor package that provides a link between cDNA microarray data stored in MAGE-ML format and the Bioconductor framework for preprocessing, visualization and analysis of microarray experiments. http://www.bioconductor.org. Open Source.


Improving the outcoupling efficiency of indium-tin-oxide-free organic light-emitting diodes via rough internal interfaces.
We present low-cost texturing methods to produce different surface roughnesses on glass substrates. Using sand blasting, abrasion and wet etching we achieve roughnesses of about 50 nm to 250 nm (root mean squared roughness Rq). These textured substrates are used as extraction elements for guided modes and substrate modes in organic light-emitting diodes (OLEDs). We evaporate 50 nm of the high index material Ta₂O₅ on the textured substrate, which acts as waveguide layer, and flatten it with the transparent photoresist SU-8. On top of that, we fabricate indium tin oxide (ITO)-free OLEDs, which are characterized by electroluminescence and photoluminescence measurements. The devices with rough interfaces obtain an up to 37.4% and 15.5% (at 20 mA/cm²) enhanced emission and it is shown that the enhancement is due to an increased outcoupling efficiency.


[We must not neglect new realities: the drama of migrants.]
Since 2014 1.8 million migrant people arrived in Europe from Italy, Spain and Greece. It represents a quarter of the London population. And other 16 thousand are the deaths in the Mediterranean Sea in four years. Yet, there are those who still talk about invasion, making it a weapon for political propaganda, and the health sector is among the most exploited. The Italian politicians themselves are at the forefront on the diffusion of fake news on the health conditions of people arriving in Europe, riding the long wave of fear of the unknown foreign people, and of the possibility of a contagion. However, data tell a completely different story: migrants who arrive in Europe does not bring vulnerability to the natives: they are vulnerable, and they need first of all our protection. There is no evidence of outbreaks of infectious diseases, and in any case border controls are carried out, and they can identify anyone who is not in good health before landing. The same can not be said of mental illness. Rape, torture, murders: this is what people arrived in Italy from Libya have lived. This should be the starting point - not the refusal to give hospitality - for aligning our communities with the UN warning from here until 2030: no one left behind.


Protein kinase C signaling and cell cycle regulation.
A link between T cell proliferation and the protein kinase C (PKC) family of serine/threonine kinases has been recognized for about 30 years. However, despite the wealth of information on PKC-mediated control of, T cell activation, understanding of the effects of PKCs on the cell cycle machinery in this cell type remains limited. Studies in other systems have revealed important cell cycle-specific effects of PKC signaling that can either positively or negatively impact proliferation. The outcome of PKC activation is highly context-dependent, with the precise cell cycle target(s) and overall effects determined by the specific isozyme involved, the timing of PKC activation, the cell type, and the signaling environment. Although PKCs can regulate all stages of the cell cycle, they appear to predominantly affect G0/G1 and G2. PKCs can modulate multiple cell cycle regulatory molecules, including cyclins, cyclin-dependent kinases (cdks), cdk inhibitors and cdc25 phosphatases; however, evidence points to Cip/Kip cdk inhibitors and D-type cyclins as key mediators of PKC-regulated cell cycle-specific effects. Several PKC isozymes can target Cip/Kip proteins to control G0/G1 → S and/or G2 → M transit, while effects on D-type cyclins regulate entry into and progression through G1. Analysis of PKC signaling in T cells has largely focused on its roles in T cell activation; thus, observed cell cycle effects are mainly positive. A prominent role is emerging for PKCθ, with non-redundant functions of other isozymes also described. Additional evidence points to PKCδ as a negative regulator of the cell cycle in these cells. As in other cell types, context-dependent effects of individual isozymes have been noted in T cells, and Cip/Kip cdk inhibitors and D-type cyclins appear to be major PKC targets. Future studies are anticipated to take advantage of the similarities between these various systems to enhance understanding of PKC-mediated cell cycle regulation in T cells.


Relationships between body temperatures and inflammation indicators under physiological and pathophysiological conditions in pigs exposed to systemic lipopolysaccharide and dietary deoxynivalenol.
We studied the constancy of the relationship between rectal and intraabdominal temperature as well as their linkage to inflammatory markers (leucocyte counts, kynurenine-to-tryptophan ratio (Kyn-Trp ratio), tumour necrosis factor alpha (TNF-α) in healthy and in pigs exposed to lipopolysaccharide (LPS) and/or deoxynivalenol (DON). Barrows (n = 44) were fed 4 weeks either a DON-contaminated (4.59 mg DON/kg feed) or a control (CON) diet and equipped with an intraabdominal temperature logger and a multicatheter system (V.portae hepatis, V.lienalis, Vv.jugulares) facilitating infusion of 0.9% NaCl (CON) or LPS (7.5 μg/kg BW) and simultaneous blood sampling. Body temperatures were measured and blood samples taken every 15 min for leucocyte counts, TNF-α and Kyn-Trp ratio. Combination of diet and infusion created six groups: CON_CONjug .-CONpor. , CON_CONjug. -LPSpor. , CON_LPSjug. -CONpor. , DON_CONjug. -CONpor. , DON_CONjug. -LPSpor. , DON_LPSjug. -CONpor. . The relationship between both temperatures was not uniform for all conditions. Linear regression revealed that an intraabdominal increase per 1°C increase in rectal temperature was ~25% higher in all LPS-infused pigs compared to NaCl-infusion, albeit diet and site of LPS infusion modified the magnitude of this difference. Inflammatory markers were only strongly present under LPS influence and showed a significant relationship with body temperatures. For example, leucocyte counts in clinically inconspicuous animals were only significantly correlated to core temperature in DON-fed pigs, but in all LPS-infused groups, irrespective of diet and temperature method. In conclusion, the gradient between body core and rectal temperature is constant in clinically inconspicuous pigs, but not under various pathophysiological conditions. In the latter, measurement of inflammatory markers seems to be a useful completion.


Model-based Meta-Analysis on the Efficacy of Pharmacological Treatments for Idiopathic Pulmonary Fibrosis.
Recently, the US Food and Drug Administration (FDA) approved the first two drugs (pirfenidone and nintedanib) indicated for the treatment of idiopathic pulmonary fibrosis (IPF). The purpose of this analysis was to leverage publicly available data to quantify comparative efficacy of compounds that are approved or in development. An analysis-ready database was developed, and the analysis dataset is composed of summary-level data from 43 arms in 20 trials, with treatment durations ranging from 8-104 weeks. A hierarchical multivariable regression model with nonparametric placebo estimation was used to fit the longitudinal profile of change from baseline of percent predicted forced vital capacity (%predicted FVC) data. Pirfenidone and nintedanib were the only drugs identified to have significant estimated positive treatment effects. Model simulations were performed to further evaluate the covariate and time course of treatment effects on longitudinal change from baseline %predicted FVC to inform future trial designs and support decision making.


Is palliative laparoscopic hyperthermic intraperitoneal chemotherapy effective in patients with malignant hemorrhagic ascites?
Malignant hemorrhagic ascites may complicate the terminal evolution of digestive cancers with peritoneal carcinomatosis. It has a bad influence on prognosis and may severely impair patients' quality of life. Palliative laparoscopic hyperthermic intraperitoneal chemotherapy (HIPEC) has been proposed to treat debilitating malignant ascites. Two cases of peritoneal carcinomatosis causing hemorrhagic ascites and severe anemia that needed iterative blood transfusions are reported. These patients were treated by laparoscopic HIPEC (mitomycin C and cisplatin with an inflow temperature of 43°C), resulting in cessation of peritoneal bleeding. No postoperative complication or relapse of ascites occurred during the following months. No more blood transfusion was needed. Laparoscopic HIPEC might be an effective and safe therapeutic option to consider in patients with malignant hemorrhagic ascites.


Reconstructing Ancient Proteins to Understand the Causes of Structure and Function.
A central goal in biochemistry is to explain the causes of protein sequence, structure, and function. Mainstream approaches seek to rationalize sequence and structure in terms of their effects on function and to identify function's underlying determinants by comparing related proteins to each other. Although productive, both strategies suffer from intrinsic limitations that have left important aspects of many proteins unexplained. These limits can be overcome by reconstructing ancient proteins, experimentally characterizing their properties, and retracing their evolution through time. This approach has proven to be a powerful means for discovering how historical changes in sequence produced the functions, structures, and other physical/chemical characteristics of modern proteins. It has also illuminated whether protein features evolved because of functional optimization, historical constraint, or blind chance. Here we review recent studies employing ancestral protein reconstruction and show how they have produced new knowledge not only of molecular evolutionary processes but also of the underlying determinants of modern proteins' physical, chemical, and biological properties.


Exporting ethics: a narrative about narrative research in South India.
The article notes some problems with the prepositional discourse of research ethics that is dominant in the West, and joins the call for an ethics-in-context approach in the human sciences. Using detailed examples from my fieldwork in South India to develop a narrative about ethical conflicts, I explore the problematics of informed consent, confidentiality and other concepts central to research ethics in the USA. The article underscores the inherent and practical risks associated with ethical universalism - applying 'universal' moral principles that have been constructed (that is, derived) in one cultural context and exporting them, without modifications, to another. The personal narrative includes my emotions in the field; they do moral work. The article draws theoretically from Bakhtin, Rabinow and feminist scholars of the Indian diaspora.


Emasculation increases seed set in the bird-pollinated hermaphrodite Kniphofia linearifolia (Xanthorrhoeaceae): evidence for sexual conflict?
Hermaphroditism in plants can lead to gender conflicts, such as pollen discounting-the loss of siring opportunities following self-pollination, and ovule discounting-the loss of seed production opportunities when self-pollen tubes disable ovules through early inbreeding depression or late-acting self-incompatibility. If ovules are discounted by self-pollination, it can be predicted that emasculation (removal of self-pollen) should increase seed production, as long as pollinators are not deterred by emasculation. • Using the hermaphroditic Kniphofia linearifolia, which belongs to a lineage known to possess late-acting self-incompatibility, we performed hand-pollinations with either self- or cross-pollen and recorded pollen tube growth and seed production. We paired experimentally emasculated plants with non-emasculated controls and quantified pollinator visitation rates, fruit, and seed set in both groups. • Ovules penetrated by tubes from self-pollen uniformly failed to develop into seeds, as expected from ovarian self-incompatibility (or strong early inbreeding depression). Experimental emasculation had no effect on the rate of visitation by birds, the primary pollinators of this species, but led to increased rejection by bees, which are secondary pollinators. Despite reducing visitation by some pollinators, emasculation led to increased seed production and reduced rates of seed abortion. • These results show that female performance in hermaphroditic flowers can be enhanced when flowers are emasculated, even if the overall number of pollinator visits is decreased. The most likely explanation for this effect of emasculation is that it reduces ovule discounting arising from pollinator-mediated self-pollination.


Induction of IgE synthesis in anti-IgM-activated nonatopic human B cells by recombinant interleukin-3.
The signals required to induce purified normal human B cell subpopulations into IgE production were studied. Pokeweed mitogen (PWM)-stimulated T cell supernatant induced IgE synthesis in low-density but not high-density Percoll-gradient-separated resting B cells. The PWM supernatant also enhanced (greater than 2-fold) IgE synthesis by anti-IgM (but not Staphylococcus A Cowan I)-activated high-density B cells (but not low-density B cells) and had affinity for lentil lectin. Subsequent studies were carried out using purified human recombinant interleukins (rIL). Human rIL-3 consistently (10/19 experiments) augmented IgE synthesis by anti-IgM-activated normal B cells. rIL-4 did not consistently (4/23 experiments) induce IgE synthesis by normal B cells or mixtures of T and B lymphocytes +/- monocytes. Furthermore, IL-1, IL-2, IL-5, IL-6, granulocyte, macrophage-colony stimulating factor (GM-CSF), granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF) and interferon-gamma also failed to induce IgE synthesis. Identification of IL-3 as the factor(s) in the PWM supernatant that was responsible for inducing IgE synthesis was inconclusive since its effect could not be reversed by the addition of anti-IL-3 antibody. Thus, our results suggest that at least two distinct soluble factors are involved in the induction of IgE synthesis by nonatopic B cells.


Complexation of bovine serum albumin and sugar beet pectin: structural transitions and phase diagram.
The complexation between bovine serum albumin (BSA) and sugar beet pectin (SBP) was studied in situ by coupling glucono-δ-lactone (GDL) induced acidification with dynamic light scattering and turbidity measurements. Individual measurements at specific pHs and mixing ratios were also carried out using zeta potentiometry, gel permeation chromatography-multiangle laser light scattering (GPC-MALLS), and isothermal titration calorimetry (ITC). These investigations together enabled the establishment of a phase diagram of BSA/SBP and the identification of the molecular events during protein/polysaccharide complexation in relation to the phase diagram, which showed five regions: (I) a stable region of mixed individual soluble polymers, (II) a stable region of intramolecular soluble complexes, (III) a quasi-stable region of intermolecular soluble complexes, (IV) an unstable region of intermolecular insoluble complexes, and (V) a second stable region of mixed individual soluble polymers, on lowering pH. We found for the first time that the complexation could take place well above the critical pH(c), the value that most previous studies had regarded as the onset occurrence of complexation. A model of structural transitions between the regions was proposed. The borderline between region II and region III represents the BSA/SBP stoichiometry for intramolecular soluble complex at a specific pH, while that between region III and region IV identifies the composition of the intermolecular insoluble complex. Also studied was the effect of NaCl and CaCl(2) on the phase diagram and structural transitions.


[Birth weight, health status and physical development of young men after 20 years].
The aim of the study was to assess the health status and physical development of young men with low birth weight (LBW) compared to normal birth weight (NBW). The study was based on the birth cohort of children born in Warsaw at six obstetric wards in 1970-74. The data from the medical records of military conscription units, concerning 682 persons with LBW and 408 persons with NBW, were taken for comparison. Mean height and weight of LBW persons were lower then for those with NBW, with similar weight-for-height proportions. The prevalence of hearing loss was higher among LBW then NBW individuals despite similar rates of ear disorders assessed at the mid-point of the follow-up. Also, neurological disorders were more prevalent among LBW then NBW individuals at the time of conscription, which could not be fully ascribed to the possible birth injuries.


Clinical status of comorbid bipolar disorder and borderline personality disorder.
The status and differentiation of comorbid borderline personality disorder and bipolar disorder is worthy of clarification. To determine whether comorbid borderline personality disorder and bipolar disorder are interdependent or independent conditions. We interviewed patients diagnosed with either a borderline personality disorder and/or a bipolar condition. Analyses of participants grouped by DSM diagnoses established that those with comorbid conditions scored similarly to those with a borderline personality disorder alone on all key variables (i.e. gender, severity of borderline personality scores, developmental stressors, illness correlates, self-injurious behaviour rates) and differed from those with a bipolar disorder alone on nearly all non-bipolar item variables. Similar findings were returned for groups defined by clinical diagnoses. Comorbid bipolar disorder and borderline personality disorder is consistent with the formal definition of comorbidity in that, while coterminous, individuals meeting such criteria have features of two independent conditions.


Assessing exposure to secondhand smoke in restaurants and bars 2 years after the smoking regulations in Beijing, China.
Field observation of patron smoking behaviors and multiple sampling approaches were conducted in 79 restaurants and bars in Beijing, 2010, 2 years after implementing the governmental smoking regulations. Smoking was observed in 30 visits to 22 of the 37 nominal non-smoking venues during peak patronage times and six visits to four of the 14 nominal non-smoking sections. The median area secondhand smoke (SHS) concentrations during peak patronage time were 27, 15, 43, and 40 μg/m(3) in nominal non-smoking venues, non-smoking sections, smoking sections, and smoking venues, respectively, as indicated by the difference between indoor and outdoor PM2.5 levels; and 1.4, 0.6, 1.7, and 2.7 μg/m(3) , respectively, as indicated by airborne nicotine. In the 27 venues with sampling of different approaches and over different time periods, the median nicotine concentration was 1.8 μg/m(3) by one-hour peak patronage-time sampling, 1.1 μg/m(3) by 1-day active area sampling, 2.5 μg/m(3) by 1-day personal sampling, and 2.3 μg/m(3) by week-long passive sampling. No significant differences in nicotine levels were observed among venues/sections with different nominal smoking policies by all sampling approaches except during peak patronage time. This study showed that the 2008 Beijing governmental smoking restriction has been poorly implemented, and SHS exposures in Beijing restaurants and bars remain high. The 2008 Beijing governmental smoking restriction requires large restaurants to designate no less than 50% of their dining area as non-smoking, without defining 'large' or specifying how the designated smoking sections and non-smoking sections should be separated. Two years after its implementation, smoking is still commonly observed in nominally non-smoking restaurants and bars and in designated non-smoking sections, and both patrons and servers are exposed to high concentrations of secondhand smoke. These results indicate that the Beijing governmental regulation fails to protect the population from SHS exposure in restaurants and bars and that more efforts are needed to pass stronger smoking regulations and ensure better compliance in Beijing, China.


Chromium infusion reverses extreme insulin resistance in a cardiothoracic ICU patient.
Insulin resistance is common and often multifactorial in acutely critically ill patients. At our institution, glycemic control is achieved in these patients using an intravenous insulin protocol. The authors present a case in which a patient developed severe insulin resistance following surgical repair of a thoracic aorta aneurysm. Postoperatively, the patient required 2110 units of insulin over 40 hours while receiving pressors and glucocorticoids. After the administration of intravenous chromium at 3 microg/h, the blood sugar normalized and insulin therapy was discontinued. This case represents a unique approach using intravenous chromium to achieve glycemic control in a patient with extreme insulin resistance and acute critical illness. Prospective clinical trials using intravenous chromium may provide the means to optimize intensive insulin therapy for critically ill patients.


Encephalitis associated with the chikungunya epidemic outbreak in Brazil: report of 2 cases with neuroimaging findings.
Chikungunya, an alphavirus infection presenting with fever, rash, and polyarthritis, is most often an acute febrile illness. Neurologic complications of chikungunya infection have been reported. Here we report the clinical and neuroimaging data of 2 patients with chikungunya-associated encephalitis during the recent Brazilian epidemic.


Anticonvulsant effect of primidone in the gerbil. Time course and significance of the active metabolites.
The anticonvulsant effect of primidone was determined in gerbils, in which seizures were elicited by a blast of compressed air, over the time range of 30 min to 18 h after oral administration. ED50s remained fairly constant from 1 to 12 h after administration: 46-73 mumol/kg with the minimal value at 6 h. Of the metabolites, phenobarbital was maximally effective at 2 h after administration (ED50 35 mumol/kg), whereas phenylethylmalondiamide (PEMA) only had a weak anticonvulsant effect (ED50 1.55 mmol/kg at 2 h). By determination of primidone and its active metabolites in plasma and brain at 1, 4 and 12 h after administration of the respective ED50s, it could be shown that unchanged primidone is mostly responsible for the anticonvulsant effect of the first hours, but, at 12 h, only phenobarbital could be detected in both tissues. PEMA could not be detected in brain. From the effective brain concentrations at different times it could be calculated that primidone and phenobarbital have the same anticonvulsant potency on a molar base in the gerbil. The concentrations necessary to control seizures in this model were considerably lower than those needed to suppress convulsions in maximal seizure models in mice and rats.


Efficacy of sulfonylureas with insulin in type 2 diabetes mellitus.
In subjects with type 2 diabetes mellitus, glycemic control deteroriates while patients use sulfonylurea drugs during the course of the disease. Adjunctive therapy with insulin at this stage requires a lesser daily insulin dose in comparison with insulin monotherapy while restoring desirable glycemic control. However, data regarding direct comparison between various sulfonylureas in this regard are lacking. To examine comparative efficacies of adjunctive therapy with insulin in subjects with type 2 diabetes manifesting lapse of glycemic control while receiving various individual sulfonylurea drugs. Four groups of 10 subjects, each presenting with glycosylated hemoglobin (HbA(1C)) >8.0% while using either tolazamide, glyburide, glipizide Gastrointestinal Therapeutic System (GITS), or glimepiride, were recruited. Two from each group were randomized to receive placebo; the others continued the same drug. Pre-supper subcutaneous 70 NPH/30 regular insulin was initiated at 10 units and gradually increased and adjusted as necessary to attain fasting blood glucose levels between 80 and 120 mg/dL and maintain the same range for 6 months. Fasting plasma glucose, plasma C-peptide, and HbA(1C) concentrations were determined prior to the addition of insulin and at the end of the study. Daily insulin dose and changes in body weight (BW) were noted at the end of the study, and the number of hypoglycemic events during the last 4 weeks of the study was determined. Daily insulin dose (units/kg BW), weight gain, and number of hypoglycemic events were significantly lower (p < 0.01) in subjects receiving sulfonylureas in comparison with placebo. However, the daily insulin dose alone was significantly lower (p < 0.05) with glimepiride (0.49 +/- 0.10; mean +/- SE) than with other sulfonylureas (tolazamide 0.58 +/- 0.12, glyburide 0.59 +/- 0.12, glipizide GITS 0.59 +/- 0.14). Finally, a significant correlation (r = 0.68; p < 0.001) was noted between suppression of plasma C-peptide level and the daily insulin dose among all participants. By lowering the daily insulin dose, sulfonylurea drugs appear to improve the sensitivity of exogenous insulin in subjects with type 2 diabetes mellitus manifesting lapse of glycemic control. Moreover, glimepiride appears to possess a greater insulin-sparing property than other sulfonylureas.


Loss of heterozygosity and mitotic linkage maps in the mouse.
Loss of heterozygosity is a significant oncogenetic mechanism and can involve a variety of mechanisms including chromosome loss, deletion, and homologous interchromosomal mitotic recombination. Analysis of H-2 antigen-loss variants from heterozygous murine cell lines provides an experimental system to estimate the relative contributions of different mechanisms for allele loss and to compare the chromosomal patterns of mitotic and meiotic recombination. Cytotoxic anti-H-2D antibodies and complement were used to isolate 161 independent target antigen-negative clones from H-2d/H-2b heterozygous cell lines; of these, 131 (84.5%) lost the allele encoding the target antigen. Allele-loss variants were typed and scored as either heterozygous or homozygous for six H-2D-proximal chromosome 17 markers and for one distal marker by restriction enzyme-site variations and Southern analysis. A single mitotic crossover could account for 50 clones (37%), with heterozygosity for at least one proximal marker and loss of heterozygosity for all markers distal to the putative recombination site. Eighty-two allele-loss variants (60%) were homozygous for all markers; the origin of these clones could be either chromosome loss or mitotic recombination between the centromere and the most proximal marker. Only 4 clones (3%) arose through more complex events such as multiple crossovers or deletion. A mitotic linkage map for mouse chromosome 17 was constructed, and the gene order deduced from somatic recombination was identical to that obtained by conventional transmission genetics. These results demonstrate that mitotic recombination is a common event leading to allele loss, in spite of the lack of evidence for frequent somatic pairing of homologous chromosomes. Mitotic mapping provides a defined system for comparison of mitotic and meiotic recombination and may lead to practical advances for elucidating somatic mechanisms of oncogenesis and for gene therapy in targeting mutations to specific sites through homologous recombination.


Occlusal traits in children with neurofibromatosis type 1.
Literature is poor of data about the occlusion in children affected by neurofibromatosis type 1 (NF1). This case-control study investigated the occlusal traits in a group of children with NF1. A hundred and fifteen children with NF1 were enrolled; non-NF1 controls were sequentially selected among subjects referred to the Pediatric Dentistry Department. All patients underwent a clinical dental examination and a panoramic radiography. The following orthodontic variables were considered: molar relationship, overjet, overbite, cross-bite, scissor bite, and crowding/spacing. Class III molar relationship resulted significantly (p = 0.01) more common in children with NF1 than in the control group as well as the unilateral posterior cross-bite (p = 0.0017). Forty-three children with NF1 (37.3%) showed radiographic abnormalities; in one case, a plexiform neurofibroma was detected. An early orthodontic evaluation might be planned in the management of children with NF1 to prevent or decrease the need for extensive orthodontic interventions.


Ivabradine Treatment Reduces Cardiomyocyte Apoptosis in a Murine Model of Chronic Viral Myocarditis.
This study was designed to explore the effects of ivabradine on cardiomyocyte apoptosis in a murine model of chronic viral myocarditis (CVMC). Mice were inoculated intraperitoneally with Coxsackievirus B3 at days 1, 14, and 28, respectively. On day 42, the mice were gavaged with ivabradine for 30 days until the 72nd day. The heart of infected mice was dilated and a large number of interstitial fibroblasts infiltrated into the myocardium on day 42. Compared with the untreated CVMC mice, mice treated with ivabradine showed a significant reduction in heart rate and less impairment of left ventricular function on day 72. The positive apoptosis of myocardial cells in the untreated CVMC group was significantly higher than that of the normal group and was significantly reduced after treatment with ivabradine. The expression levels of Bax and Caspase-3 in the untreated CVMC group were significantly higher than those of the normal group and were apparently reduced in the ivabradine-treated group versus the untreated CVMC group. Bcl-2 showed a high expression in the normal group and low expression in the untreated CVMC group, but its expression level in the ivabradine-treated group were higher than that of the untreated CVMC group. These results indicate that ivabradine could attenuate the expression of Caspase-3 by downregulation of Bax and upregulation of Bcl-2 to prevent the deterioration of cardiac function resulting from ventricular myocyte loss by cardiomyocyte apoptosis.


Effects of gibberellins on important agronomic traits of horticultural plants.
Horticultural plants such as vegetables, fruits, and ornamental plants are crucial to human life and socioeconomic development. Gibberellins (GAs), a class of diterpenoid compounds, control numerous developmental processes of plants. The roles of GAs in regulating growth and development of horticultural plants, and in regulating significant progress have been clarified. These findings have significant implications for promoting the quality and quantity of the products of horticultural plants. Here we review recent progress in determining the roles of GAs (including biosynthesis and signaling) in regulating plant stature, axillary meristem outgrowth, compound leaf development, flowering time, and parthenocarpy. These findings will provide a solid foundation for further improving the quality and quantity of horticultural plants products.


The paired neuroglial and interglial membranes in the crayfish stretch receptor and their local disorganization.
The paired neuronal and glial membranes, or interglial membranes, which are separated by the narrow layer of the extracellular medium, are involved in intercellular communications. In the crayfish stretch receptor, the paired neuroglial membranes contain thin protein bridges (septate junctions) that maintain the intermembrane gap. In some places the paired membranes are locally disorganized. In the altered regions, they comprise the diffuse material in which a few 10-15 nm vesicles are embedded. The development of these defects can lead to formation of 20-30 nm vesicles and perforations in the paired membranes. The presence of such holes can, in principle, disturb ionic gradients and neuronal activity. However, a free passage between contacting neurons and glia is prevented by the diffuse proteolipid material (the product of the membrane disorganization) that seals perforations. As a result, the neuroglial border does not lose its integrity and impermeability for ions so that the sensory neurons save the capability for prolonged regular firing. Unlike the neuroglial border, some perforations in the paired glia-glial membranes are not sealed. This can create the interglial syncytial connections providing the shortcut pathway for transport of ions and metabolites across the glial layers in the crayfish stretch receptor.


Studies of the responses of basophil and eosinophil leucocytes and mast cells to the nematode Trichostrongylus colubriformis. I. Observations during the expulsion of first and second infections by guinea-pigs.
During expulsion by guniea-pigs of both first and second infections with the intestinal nematode parasite Trichostrongylus colubriformis, increased numbers of basophils and eosinophils were present in the bone marrow and blood. In addition, increased numbers of basophils, mast cells and eosinophils, many possessing reduced numbers of cytoplasmic granules, accumulated at the sites of infection. Blood eosinophil counts, however, did not show significant changes during the early part of a first infection.


Analysis of translation elongation factors from wheat during development and following heat shock.
Translational activity in plants undergoes rapid changes during developmental stages such as seed formation and germination, and during abiotic stresses such as heat shock, hypoxia and wounding. We examined the protein levels and isoelectric state of two components of the translation machinery, elongation factor (EF) 1 alpha and 2, to determine their roles in the regulation of translation. We found that the apparent protein levels of EF1 alpha increase relative to the EF2 levels which decline slightly during the development of the wheat seed. During germination, high levels of these factors are present in seedling tissues known to be actively engaged in translation; however, no differences in isoelectric state were observed during germination. As an example of abiotic stress, heat shock had little impact on the apparent levels of EF1 alpha or EF2 present in wheat leaves, nor were changes in the number or levels of isoforms observed.


[Vasovagal syncope in pediatric patients: a medium-term follow-up analysis].
Little information is available on the evolution of pediatric patients with vasovagal syncope. We therefore aimed to assess the medium-term clinical outcome of children evaluated by tilt testing for syncope of unknown origin. Fifty-one children under 17 years of age who had undergone tilt testing were identified from a data base and studied prospectively. Kaplan-Meier and Cox regression analyses were performed to estimate syncope-free survival, its predictors, and the relative risks of several patient subgroups. Forty-seven (92%) of the children were followed for a mean 21 9 months. The rate of recurrence of syncope was considerably lower than that estimated during history taking before the tilt test (19% vs 47%; p < 0.01). Although the low rate made it difficult to identify predictors, several potential predictors emerged from the multivariate analysis. Only the history of more than one syncope before the tilt test (vs. isolated syncope) was found to have independent predictive value (p = 0.04). The cumulative probability of recurrence projected for a period of 38 months was 66.2% (SEM = 16.5%) for children with more than one syncope before testing vs. 0% for those who had experienced only one. No other events occurred. The medium-term prognosis seems to be good for children with vasovagal syncope of unknown origin, given the low rate of recurrence, regardless of the results of tilt testing. The only predictor of recurrent syncope was pretest history, such that children with only one syncope before testing experience no recurrence and those with one or more episodes are estimated to have an increasingly higher likelihood of recurrence. These data may be useful for the recommending tilt testing and for planning therapy for children with vasovagal syncope.


Effects of deoxyadenosine triphosphate and 9-beta-D-arabinofuranosyl-adenine 5'-triphosphate on human ribonucleotide reductase from Molt-4F cells and the concept of "self-potentiation".
Deoxyadenosine triphosphate (dATP) acted as a noncompetitive inhibitor with respect to the specific nucleoside triphosphate activator for the reduction of all four common ribonucleoside diphosphates catalyzed by the reductase derived from human Molt-4F (T-type lymphoblast) cells. The inhibition constant of dATP for different ribonucleotide reduction reactions was different, indicating that the binding of the nucleoside triphosphate activator or substrate could modify the binding affinity of dATP to the enzyme. dATP also acted as a noncompetitive inhibitor with respect to cytidine diphosphate (CDP) for reductase-catalyzed CDP reduction. 9-beta-D-Arabinofuranosyl-adenine 5'-triphosphate acted as a competitive inhibitor with respect to either adenosine triphosphate or guanosine triphosphate for CDP or for adenosine diphosphate reduction, respectively. The inhibition constant was 15 microM for CDP reduction and 4 microM for adenosine diphosphate reduction. 1-beta-D-Arabinofuranosyladenine 5'-triphosphate could not substitute for adenosine triphosphate or guanosine triphosphate as the activator for CDP or adenosine diphosphate reduction, respectively. The effects of 9-beta-D-arabinofuranosylcytosine 5'-triphosphate and 5-iodo-2'-deoxyuridine 5'-triphosphate on ribonucleotide reductase were also included for comparison. The "self-potentiation" mechanism of the action of 9-beta-D-arabinofuranosyladenine and 5-iodo-2'-deoxyuridine is discussed.


First-Trimester Urinary Bisphenol A Concentration in Relation to Anogenital Distance, an Androgen-Sensitive Measure of Reproductive Development, in Infant Girls.
Evidence from animal models suggests that prenatal exposure to bisphenol A (BPA), a ubiquitous endocrine-disrupting chemical, is associated with adverse reproductive outcomes in females. Exposure during early gestation, a critical period for reproductive development, is of particular concern. Anogenital distance (AGD) is a sensitive biomarker of the fetal hormonal milieu and a measure of reproductive toxicity in animal models. In some studies, the daughters of BPA-exposed dams have shorter AGD than controls. Here, we investigate this relationship in humans. BPA was assayed in first-trimester urine samples from 385 participants who delivered infant girls in a multicenter pregnancy cohort study. After birth, daughters underwent exams that included two measures of AGD (AGD-AC: distance from center of anus to clitoris; AGD-AF: distance from center of anus to fourchette). We fit linear regression models to examine the association between specific gravity-adjusted (SPG-adj) maternal BPA concentrations and infant AGD, adjusting for covariates. BPA was detectable in 94% of women. In covariate-adjusted models fit on 381 eligible subjects, the natural logarithm of SpG-adj maternal BPA concentration was inversely associated with infant AGD-AC [β=−0.56, 95% confidence interval (CI): −0.97, −0.15]. We observed no association between maternal BPA and infant AGD-AF. BPA may have toxic effects on the female reproductive system in humans, as it does in animal models. Higher first-trimester BPA exposure was associated with significantly shorter AGD in daughters, suggesting that BPA may alter the hormonal environment of the female fetus. https://doi.org/10.1289/EHP875.


Emerging platforms using liquid biopsy to detect EGFR mutations in lung cancer.
Advances in target therapies for lung cancer have enabled detection of gene mutations, specifically those of EGFR. Assays largely depend on the acquisition of tumor tissue biopsy, which is invasive and may not reflect the genomic profile of the tumor at treatment due to tumor heterogeneity or changes that occur during treatment through acquired resistance. Liquid biopsy, a blood test that detects evidence of cancer cells or tumor DNA, has generated considerable interest for its ability to detect EGFR mutations. However, its clinical application is limited by complicated collection methods and the need for technique-dependent platforms. Recently, simpler techniques for EGFR mutant detection in urine or saliva samples have been developed. This review focuses on advances in liquid biopsy and discusses its potential for clinical implementation in lung cancer.


Multinodular goiter management in Western Saudi Arabia.
The aim of this study was to provide epidemiological data and summarize the different modalities of management of multinodular goiters (MNG) in the Western Province of Saudi Arabia (KSA). A total of 135 patients had MNG, which were diagnosed by ultrasonography at King Abdul-Aziz University Hospital in Jeddah, KSA between January 2003 and June 2004. The data collected includes the age, gender, physical examination of the thyroid gland (trifluorothymidine, free thyroxine, free triiodothyronine, thyroid stimulating hormone), ultrasonographic findings and Tc99 radioiodine nuclear scan of the thyroid gland, fine needle aspiration (FNA) of the gland and antithyroid antibodies. The study group was divided according to the thyroid status: nontoxic euthyroid MNG, hypothyroid MNG and toxic hyperthyroid MNG. The management of MNGs according to the patients' clinical presentations and thyroid status was reviewed. The mean age was 39 +/- 12.66 years with a range of 10-79 years. Forty-two patients (31%), with no history of thyroid cancer, had ultrasoundguided FNA; 41 (97.6%) of the FNAs were benign. One FNA (2.3%) was positive for papillary carcinoma in a 56-year-old female patient. Thyroid antibodies (anti-microsomal antibodies and anti-thymoglobulin antibodies) were measured in 50 patients (37%). Thirty-one (62%) were positive, 11 (35%) of which were positive in patients with documented hypothyroidism. All patients with hypothyroidism were treated with levothyroxine; however, only 25.6% with euthyroid MNG were treated with suppressive doses of levothyroxine. Twelve patients (44.4%) with toxic MNG were treated with antithyroid medications. Radioiodine therapy was not given to any patient with nontoxic MNG; however, 48% of those with hyperthyroid MNG received radioactive iodine treatment. Surgery was carried out in 25.6% of patients with euthyroid nontoxic MNG, and in 11 patients with toxic MNG. Nearly half of those with nontoxic MNG (46.5%) and 14.8% of those with a subclinical hyperthyroid MNG refrained from any therapy. They were followed-up by their clinician in the outpatient clinic. Serum thyroid stimulating hormone levels, ultrasound and fine needle biopsy were the cornerstones of the diagnostic evaluation of patients with MNG. Review of the management of these patients was comparable to that found in the literature. The treatment strategy was similar to the recommendations by the American Thyroid Society; however, radioactive iodine treatment was not used as a treatment for patients with nontoxic goiters.


MethyCancer: the database of human DNA methylation and cancer.
Cancer is ranked as one of the top killers in all human diseases and continues to have a devastating effect on the population around the globe. Current research efforts are aiming to accelerate our understanding of the molecular basis of cancer and develop effective means for cancer diagnostics, treatment and prognosis. An altered pattern of epigenetic modifications, most importantly DNA methylation events, plays a critical role in tumorigenesis through regulating oncogene activation, tumor suppressor gene silencing and chromosomal instability. To study interplay of DNA methylation, gene expression and cancer, we developed a publicly accessible database for human DNA Methylation and Cancer (MethyCancer, http://methycancer.genomics.org.cn). MethyCancer hosts both highly integrated data of DNA methylation, cancer-related gene, mutation and cancer information from public resources, and the CpG Island (CGI) clones derived from our large-scale sequencing. Interconnections between different data types were analyzed and presented. Furthermore, a powerful search tool is developed to provide user-friendly access to all the data and data connections. A graphical MethyView shows DNA methylation in context of genomics and genetics data facilitating the research in cancer to understand genetic and epigenetic mechanisms that make dramatic changes in gene expression of tumor cells.


Perception of illness in patients with traumatic brain injury.
Perception of illness plays an important role in recovery process. It affects our coping behaviors, adherence to treatment and preventive measures taken for healthy recovery. The aim of the study is to examine perception of illness in patients with traumatic brain injury (TBI). This was a cross sectional study design done on sample of 31 patients with mild to moderate TBI. Depression anxiety stress scales-21 (DASS-21), Brief illness perception questionnaire (IPQ) and Rivermead Post Concussion Symptoms Questionnaire (RPQ) was used. Descriptive and correlational statistics was used. The results indicated that overall higher percentage falls within low and medium range of IPQ. However higher percentage falls within higher range, on coherence and emotional response subscales of IPQ. Consequence, timeline, personal control, treatment control, concern, emotional control, and total of the subscales of IPQ were positively correlated with RPQ3 and RPQ13 at 0.01 and 0.05 level of significance. A significant correlation was found between demographic variables and subscales of IPQ at 0.01 and 0.05 level of significance. The study shows one to one relationship between symptoms experienced by patients, how they perceive their illness and socio demographic variables.


Up-scaling single cell-inoculated suspension culture of human embryonic stem cells.
We have systematically developed single cell-inoculated suspension cultures of human embryonic stem cells (hESC) in defined media. Cell survival was dependent on hESC re-aggregation. In the presence of the Rho kinase inhibitor Y-27632 (Ri) only approximately 44% of the seeded cells were rescued, but an optimized heat shock treatment combined with Ri significantly increased cell survival to approximately 60%. Mechanistically, our data suggest that E-cadherin plays a role in hESC aggregation and that dissociation and re-aggregation upon passaging functions as a purification step towards a pluripotency markers-enriched population. Mass expansion of hESC was readily achieved by up-scaling 2 ml cultures to serial passaging in 50 ml spinner flasks. A media comparison revealed that mTeSR was superior to KnockOut-SR in supporting cell proliferation and pluripotency. Persistent expression of pluripotency markers was achieved for two lines (hES2, hES3) that were used at higher passages (>86). In contrast, rapid down regulation of Oct4, Tra-1-60, and SSEA4 was observed for ESI049, a clinically compliant line, used at passages 20-36. The up-scaling strategy has significant potential to provide pluripotent cells on a clinical scale. Nevertheless, our data also highlights a significant line-to-line variability and the need for a critical assessment of novel methods with numerous relevant cell lines.


Tuning the Elasticity of Polymersomes for Brain Tumor Targeting.
Nanoformulations show great potential for delivering drugs to treat brain tumors. However, how the mechanical properties of nanoformulations affect their ultimate brain destination is still unknown. Here, a library of membrane-crosslinked polymersomes with different elasticity are synthesized to investigate their ability to effectively target brain tumors. Crosslinked polymersomes with identical particle size, zeta potential and shape are assessed, but their elasticity is varied depending on the rigidity of incorporated crosslinkers. Benzyl and oxyethylene containing crosslinkers demonstrate higher and lower Young's modulus, respectively. Interestingly, stiff polymersomes exert superior brain tumor cell uptake, excellent in vitro blood brain barrier (BBB) and tumor penetration but relatively shorter blood circulation time than their soft counterparts. These results together affect the in vivo performance for which rigid polymersomes exerting higher brain tumor accumulation in an orthotopic glioblastoma (GBM) tumor model. The results demonstrate the crucial role of nanoformulation elasticity for brain-tumor targeting and will be useful for the design of future brain targeting drug delivery systems for the treatment of brain disease.


A study of the specificities of sequential antisera to variola and monkeypox viruses by radioimmunoassay.
The specificities of antisera during development of the humoral antibody response to poxvirus antigens were examined in monkeys injected with chimp-9 whitepox virus or monkeypox virus. Sera were obtained from 3 African green (vervet) monkeys inoculated with chimp-9 whitepox virus, 1 rhesus monkey inoculated with monkeypox virus, and 2 rhesus monkeys inoculated with soluble monkeypox viral antigen. The sequentially obtained sera from each animal were adsorbed with uninfected chicken chorioallantoic membranes (CAM) or vaccinia virus-infected CAM. The adsorbed sera were tested by radioimmunoassay to determine the specificity of the residual antibodies to vaccinia, variola, and monkeypox viruses. The adsorbed sera at different stages of the immune response showed increasing specificity with time after inoculation. Generally, antibodies in sera collected earlier than 21-27 days after immunization could not be identified after adsorption, but late sera could be identified unequivocally.


Fetal Limb Ischaemia in Twin-to-Twin Transfusion Syndrome.
Objective. To describe the rare association between prenatal vascular limb occlusion and twin-to-twin transfusion syndrome. The Case. A woman with severe twin-to-twin transfusion syndrome was treated with fetoscopic laser ablation at 19-week gestation. At 27 weeks, the twins were delivered by an emergency caesarean section. The right arm of twin 1, the recipient twin, was noted to be "ischaemic" and was later amputated. Conclusion. This case is unusual in that it affected the upper limb and there was no evidence of polycythaemia, which is a suggested pathological mechanism. It was initially thought that the limb damage was due to the laser ablation, but after discussion with the fetal medicine team vascular limb occlusion in association with twin-to-twin transfusion syndrome was considered. Limb ischaemia is a serious complication of twin-to-twin transfusion syndrome and is unrelated to any form of fetal therapy. Implications. Neonatologists and paediatricians need to be aware of this association as it has medicolegal implications and parents should be counselled as to the possible, albeit rare, occurrence, especially when twin-to-twin transfusion syndrome is of advanced stage at presentation.


Assessment of candidemia-attributable mortality in critically ill patients using propensity score matching analysis.
Candidemia in critically ill patients is usually a severe and life-threatening condition with a high crude mortality. Very few studies have focused on the impact of candidemia on ICU patient outcome and attributable mortality still remains controversial. This study was carried out to determine the attributable mortality of ICU-acquired candidemia in critically ill patients using propensity score matching analysis. A prospective observational study was conducted of all consecutive non-neutropenic adult patients admitted for at least seven days to 36 ICUs in Spain, France, and Argentina between April 2006 and June 2007. The probability of developing candidemia was estimated using a multivariate logistic regression model. Each patient with ICU-acquired candidemia was matched with two control patients with the nearest available Mahalanobis metric matching within the calipers defined by the propensity score. Standardized differences tests (SDT) for each variable before and after matching were calculated. Attributable mortality was determined by a modified Poisson regression model adjusted by those variables that still presented certain misalignments defined as a SDT > 10%. Thirty-eight candidemias were diagnosed in 1,107 patients (34.3 episodes/1,000 ICU patients). Patients with and without candidemia had an ICU crude mortality of 52.6% versus 20.6% (P < 0.001) and a crude hospital mortality of 55.3% versus 29.6% (P = 0.01), respectively. In the propensity matched analysis, the corresponding figures were 51.4% versus 37.1% (P = 0.222) and 54.3% versus 50% (P = 0.680). After controlling residual confusion by the Poisson regression model, the relative risk (RR) of ICU- and hospital-attributable mortality from candidemia was RR 1.298 (95% confidence interval (CI) 0.88 to 1.98) and RR 1.096 (95% CI 0.68 to 1.69), respectively. ICU-acquired candidemia in critically ill patients is not associated with an increase in either ICU or hospital mortality.


Dissecting aortic aneurysm in pregnant women without Marfan disease.
Dissection of the young women without Marfan disease is related, in most instances, to pregnancy. The structural changes in the arterial wall that occur during pregnancy may predispose women to this complication of pregnancy. In this study, two young women, who had none of the characteristics of Marfan's syndrome, were operated upon for dissection of the ascending aorta. The first patient was in the 35th week of gestation and a cesarean section was performed prior to replacement of the ascending aorta. In the second patient, diagnosis of dissection and severe aortic regurgitation occurred in the postpartum period. Knowledge of this albeit relatively rare complication of pregnancy may assist the clinician in diagnosing and referring patients early for surgical treatment that, in most instances, may save the lives of both mother and fetus.


Vasopressin-stimulated increase in phosphorylation at Ser269 potentiates plasma membrane retention of aquaporin-2.
Vasopressin controls water excretion through regulation of aquaporin-2 (AQP2) trafficking in renal collecting duct cells. Using mass spectrometry, we previously demonstrated four phosphorylated serines (Ser256, Ser261, Ser264, and Ser269) in the carboxyl-terminal tail of rat AQP2. Here, we used phospho-specific antibodies and protein mass spectrometry to investigate the roles of vasopressin and cyclic AMP in the regulation of phosphorylation at Ser269 and addressed the role of this site in AQP2 trafficking. The V2 receptor-specific vasopressin analog dDAVP increased Ser(P)269-AQP2 abundance more than 10-fold, but at a rate much slower than the corresponding increase in Ser256 phosphorylation. Vasopressin-mediated changes in phosphorylation at both sites were mimicked by cAMP addition and inhibited by protein kinase A (PKA) antagonists. In vitro kinase assays, however, demonstrated that PKA phosphorylates Ser256, but not Ser269. Phosphorylation of AQP2 at Ser269 did not occur when Ser256 was replaced by an unphosphorylatable amino acid, as seen in both S256L-AQP2 mutant mice and in Madin-Darby canine kidney cells expressing an S256A mutant, suggesting that Ser269 phosphorylation depends upon prior phosphorylation at Ser256. Immunogold electron microscopy localized Ser(P)269-AQP2 solely in the apical plasma membrane of rat collecting duct cells, in contrast to the other three phospho-forms (found in both apical plasma membrane and intracellular vesicles). Madin-Darby canine kidney cells expressing an S269D "phosphomimic" AQP2 mutant showed constitutive localization at the plasma membrane. The data support a model in which vasopressin-mediated phosphorylation of AQP2 at Ser269:(a) depends on prior PKA-mediated phosphorylation of Ser256 and (b) enhances apical plasma membrane retention of AQP2.


Learning effect in automatic static perimetry with Dicon TKS 4000 computed perimeter.
To find out the significance of the learning effect in repeated computed perimetry. We performed computed perimetry twice within one week period on 35 eyes (20 patients) with suspected glaucoma and visual acuity > 0.5 (20/40). The program used was with 30 degrees threshold grid. For each quadrant of the visual field were recorded the mean deviation readings. These readings were compared for the initial and the second perimeters for each eye. The mean deviation from the normal readings were significantly higher for the initial perimetric exam for all quadrants (Wilcoxon test for matched pairs, p < 0.05). The visual field defects, if present, were more concentrated and with sharper borders in the second exams. This study suggests that there is a significant learning effect in computed perimetry. A visual field exam, showing defects suspicious for glaucoma, should be repeated within one week and the second one taken for further referral.


Interaction of bacterial genera associated with therapeutic response to immune checkpoint PD-1 blockade in a United States cohort.
Recent studies show that human gut microbial composition can determine whether a patient is a responder or non-responder to immunotherapy but have not identified a common microbial signal shared by responding patients. The functional relationship between immunity, intestinal microbiota, and NSCLC response to immune checkpoint inhibitor/inhibition (ICI) in an American cohort remains unexplored. RNAlater-preserved fecal samples were collected from 65 pre-treatment (baseline) and post-treatment stage III/IV NSCLC patients undergoing ICI therapy, categorized as responders or non-responders according to RECIST criteria. Pooled and individual responder and non-responder microbiota were transplanted into a gnotobiotic mouse model of lung cancer and treated with ICIs. 16S rDNA and RNA sequencing was performed on patient fecal samples, 16S rDNA sequencing on mouse fecal samples, and flow cytometric analysis on mouse tumor tissue. Responder patients have both a different microbial community structure than non-responders (P = 0.004) and a different bacterial transcriptome (PC2 = 0.03) at baseline. Taxa significantly enriched in responders include amplicon sequence variants (ASVs) belonging to the genera Ruminococcus, Akkermansia, and Faecalibacterium. Pooled and individual responder microbiota transplantation into gnotobiotic mice decreased tumor growth compared to non-responder colonized mice following ICI (P = 0.023, P = 0.019, P = 0.008, respectively). Responder tumors showed an increased anti-tumor cellular phenotype following ICI treatment. Responder mice are enriched with ASVs belonging to the genera Bacteroides, Blautia, Akkermansia, and Faecalibacterium. Overlapping taxa mapping between human and mouse cohorts correlated with tumor size and weight revealed a network highlighting responder-associated ASVs belonging to the genera Colidextribacter, Frisingicoccus, Marvinbryantia, and Blautia which have not yet been reported. The role of isolate-specific function and bacterial gene expression in gut microbial-driven responsiveness to ICI has been underappreciated. This work supports further investigation using isolate-driven models to characterize the mechanisms underlying this phenomenon.


Performance of the VITEK MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system for rapid bacterial identification in two diagnostic centres in China.
Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS systems was not officially launched for diagnostic use in clinical microbiology laboratories in China until 2012. Here, we report the findings from the first large-scale evaluation study of VITEK MS for routine bacterial identification in two major diagnostic centres in Beijing and Hong Kong. A total of 2266 unique isolates representing 56 genera and 127 species were analysed, and results were compared to those obtained by VITEK 2. Any discrepancies were resolved by 16S rRNA sequencing. Overall, VITEK MS provided correct identification for 2246 (99.1%) isolates, including 2193 (96.8 %) with correct species-level identifications and 53 (2.3 %) matched at the genus level only. VITEK MS surpassed VITEK 2 consistently in species-level identification of important pathogens, including non-Enterobacteriaceae Gram-negative bacilli (94.7 versus 92 %), staphylococci (99.7 versus 92.4 %), streptococci (92.6 versus 79.4 %), enterococci (98.8 versus 92.6 %) and Clostridium spp. (97.3 versus 55.5 %). The findings demonstrated that VITEK MS is highly accurate and reliable for routine bacterial identification in clinical settings in China.


Synthesis of new acyclic pyrimidine nucleoside analogs as potential antiviral drugs.
The synthesis of 6-[N-(4-hydroxybutyl)amino]pyrimidinone derivatives 18-23 and the acyclic phosphonate nucleoside analogs 29-30 is reported. Their cytotoxic and antiviral effects were investigated. 2,5-Diamino-6-[N-[2-(phosphonomethoxy)ethyl]amino]pyrimidin-4(3H)- one (30) showed strong antiviral effects, and 21 showed significant cytotoxic activity.


Mechanisms of the depot specificity of peroxisome proliferator-activated receptor gamma action on adipose tissue metabolism.
In this study, we aimed to establish the mechanisms whereby peroxisome proliferator-activated receptor gamma (PPARgamma) agonism brings about redistribution of fat toward subcutaneous depots and away from visceral fat. In rats treated with the full PPARgamma agonist COOH (30 mg x kg(-1) x day(-1)) for 3 weeks, subcutaneous fat mass was doubled and that of visceral fat was reduced by 30% relative to untreated rats. Uptake of triglyceride-derived nonesterified fatty acids was greatly increased in subcutaneous fat (14-fold) and less so in visceral fat (4-fold), with a concomitant increase, restricted to subcutaneous fat only, in mRNA levels of the uptake-, retention-, and esterification-promoting enzymes lipoprotein lipase, aP2, and diacylglycerol acyltransferase 1. Basal lipolysis and fatty acid recycling were stimulated by COOH in both subcutaneous fat and visceral fat, with no frank quantitative depot specificity. The agonist increased mRNA levels of enzymes of fatty acid oxidation and thermogenesis much more strongly in visceral fat than in subcutaneous fat, concomitantly with a stronger elevation in O2 consumption in the former than in the latter. Mitochondrial biogenesis was stimulated equally in both depots. These findings demonstrate that PPARgamma agonism redistributes fat by stimulating the lipid uptake and esterification potential in subcutaneous fat, which more than compensates for increased O2 consumption; conversely, lipid uptake is minimally altered and energy expenditure is greatly increased in visceral fat, with consequent reduction in fat accumulation.


Utility of frozen section analysis of resection margins during partial nephrectomy.
To evaluate the utility of routine intraoperative frozen-section histologic analysis during partial nephrectomy to ensure negative surgical margins. Partial nephrectomy has gained acceptance for surgical treatment of small renal cancers. Many surgeons send specimens for intraoperative frozen section histologic analysis to ensure negative margins. We reviewed the records of 78 patients who underwent partial nephrectomy for presumed malignancy. Patient demographics, intraoperative findings, and pathologic and clinical outcomes were analyzed. Seventy-nine partial nephrectomies were performed in 78 patients. Frozen sections were obtained intraoperatively in 76 cases. In 1 case (1.3%), a single margin was interpreted as positive for carcinoma, prompting deeper resection. The final histopathologic finding was interpreted as angiomyolipoma rather than carcinoma. The final pathologic examination revealed renal cell carcinoma in 52 (66%) of 79 cases. The mean oncologic follow-up was 16.2 months. One local recurrence was noted (1.9%). It arose in the resection bed 19 months after removal of a 4.5-cm tumor (pathologic Stage T3a). Both intraoperative frozen section margins and final pathologic margins were negative in this case. One patient developed pulmonary metastases and represented the only metastatic recurrence, as well as the only cancer-related death in our cohort (1.9%). Our data suggest that when partial nephrectomy is performed with attention to excising a perimeter of grossly normal-appearing parenchyma, sending specimens for intraoperative frozen section analyses may provide an unnecessary expense without providing meaningful, reliable information. Additional studies, including larger cohorts and extended follow-up, are needed to support these results.


Multinational cost-effectiveness analysis of empagliflozin for heart failure patients with ejection fraction >40.
Heart failure is a chronic progressive condition, with considerable burden on patients' quality of life and economic burden for the healthcare systems. Before the approval of empagliflozin, there were no proven effective treatments for patients with heart failure with left ventricular ejection fraction (HF LVEF) > 40%. The aim of this study was to evaluate the cost-effectiveness of empagliflozin + standard of care (SoC) compared with SoC alone for patients with HF LVEF > 40%, from the perspective of the healthcare systems of the United Kingdom (UK), Spain, and France, and to quantify the healthcare costs for these patients. A lifetime Markov cohort state-transition model was developed based on discrete health states defined by Kansas City Cardiomyopathy Questionnaire-Clinical Summary Score quartiles to track disease severity. Model inputs relied primarily on the EMPEROR-Preserved trial data or obtained from published literature or country-specific databases, as well as local guidelines for the requirements for the conduct of the economic evaluation of healthcare technologies. The total lifetime cost of receiving SoC per patient was £10 092, €15 765, and €14 958 in the UK, Spain, and France, respectively, which increased by £1407, €1148, and €1485, respectively, with the addition of empagliflozin to the SoC. Empagliflozin + SoC was associated with significantly reduced number of hospitalization for HF or cardiovascular death compared with SoC alone, which was a key driver offsetting its drug acquisition costs. The incremental cost-effectiveness ratio per quality-adjusted life year (QALY) gained was consistently favourable at £14 851, €11 706, and €15 447 in the UK, Spain, and France, respectively. Scenario analysis using the New York Heart Association functional class showed similar results. Probabilistic sensitivity analyses showed more than 50% probability for cost-effectiveness for a willingness-to-pay (WTP) threshold of £/€20 000/QALY for the three countries. Empagliflozin was found to be the first targeted treatment option that is clinically effective and cost-effective for patients with HF LVEF > 40%. Prescribing empagliflozin with SoC to patients with HF LVEF > 40% is expected to improve clinical outcomes and patients' quality of life and substantially below accepted WTP threshold for the healthcare systems in the UK, Spain, and France.


Determination of chlorate, perchlorate and bromate anions in water samples by microbore reversed-phase liquid chromatography coupled to sonic-spray ionization mass spectrometry.
Sonic-spray ionization mass spectrometry (SSI-MS) has recently been shown to provide similar mass spectra to those generated by electrospray ionization mass spectrometry for a wide range of compounds, i.e. from small inorganic species to peptides, proteins and numerous other biomolecules. However, limited information about this new ionization technique, such as sensitivity, limit of detection and quantification accuracy, has been reported. In particular, its coupling to liquid chromatography needs further development and assessment, along with the introduction of a broad range of applications. A high-efficiency glass pneumatic nebulizer, used for decades for sample introduction in atomic spectrometry, was used for the SSI-MS analysis of chlorate (ClO3- ), perchlorate (ClO4- ) and bromate (BrO3- ) anions, following their separation using reversed-phase microbore high-performance liquid chromatography and tandem mass spectrometry (MS/MS) operated in selected reaction monitoring mode. The developed and optimized microbore HPLC/SSI-MS/MS technique exhibited low limits of detection: 5.3 ng L-1 for chlorate, 10 ng L-1 for perchlorate and 33.7 ng L-1 for bromate, and provided reliable and accurate measurements of chlorate concentrations in water samples as demonstrated when comparing it with Ion Chromatography-Conductivity Detection (IC-CD), the benchmark technique for ion quantitation. This is the first time that the use of HPLC/SSI-MS/MS has been reported for the detection and quantitation of chlorate, perchlorate and bromate in water samples. In addition, the exceptionally low LODs achieved using SSI render the technique competitive with the established and dominating electrospray ionization technique. Here, we have demonstrated that a commercially available high-efficiency glass pneumatic nebulizer can also be used, without any further modification, as an efficient gas-phase ion source. Copyright © 2017 John Wiley & Sons, Ltd.


Inflammatory potential of diet and risk of cardiovascular disease or mortality: A meta-analysis.
Inconsistent findings have reported on the inflammatory potential of diet and cardiovascular disease (CVD) and mortality risk. The aim of this meta-analysis was to investigate the association between the inflammatory potential of diet as estimated by the dietary inflammatory index (DII) score and CVD or mortality risk in the general population. A comprehensive literature search was conducted in PubMed and Embase databases through February 2017. All prospective observational studies assessing the association of inflammatory potential of diet as estimated by the DII score with CVD and all-cause, cancer-related, cardiovascular mortality risk were included. Nine prospective studies enrolling 134,067 subjects were identified. Meta-analyses showed that individuals with the highest category of DII (maximal pro-inflammatory) was associated with increased risk of all-cause mortality (hazard risk [HR] 1.22; 95% confidence interval [CI] 1.06-1.41), cardiovascular mortality (RR 1.24; 95% CI 1.01-1.51), cancer-related mortality (RR 1.28; 95% CI 1.04-1.58), and CVD (RR 1.32; 95% CI 1.09-1.60) than the lowest DII score. More pro-inflammatory diets, as estimated by the higher DII score are independently associated with an increased risk of all-cause, cardiovascular, cancer-related mortality, and CVD in the general population, highlighting low inflammatory potential diet may reduce mortality and CVD risk.


Determination of daidzein and genistein contents in mangifera fruit.
The aim of this study was to determine the daidzein and genistein contents in Mangifera fruits. Three Mangifera species namely 'bacang' (Mangifera foetida), 'kuini' (M. odorata) and 'bambangan' (M. pajang) each from two different locations were selected. The extraction of isoflavones was carried out at 80oC for 30, 60 and 90 min. HPLC method was performed with a flow rate of 1.00 ml/min using three different separation columns to determine isoflavone contents. The Zorbax Eclipse RP C18 reverse-phase column was found to give the best resolution for isoflavone separation in Mangifera fruits. Moreover, extraction time of 90 min was found to increase the isoflavone aglycone contents. At optimised condition, 'kuini'' had relatively high daidzein (9.4-10.5 mg/100 g) and genistein (1.6-1.7 mg/100 g) contents. Daidzein content of 'bambangan' (8.3-8.7 mg/100 g) was higher than 'bacang', but the genistein content of 'bambangan' (0.4-0.6 mg/100 g) was similar to that of 'bacang' (0.4-0.8 mg/100 g). However, there was a variation in daidzein and genistein contents in Mangifera fruits between two geographical locations.


The Effect of Aging on Relationships between Lean Body Mass and VO2max in Rowers.
Aging is associated with a fall in maximal aerobic capacity as well as with a decline in lean body mass. The purpose of the study was to investigate the influence of aging on the relationship between aerobic capacity and lean body mass in subjects that chronically train both their upper and lower bodies. Eleven older rowers (58±5 yrs) and 11 younger rowers (27±4 yrs) participated in the study. Prior to the VO2max testing, subjects underwent a dual energy X-ray absorptiometry scan to estimate total lean body mass. Subsequently, VO2max was quantified during a maximal exercise test on a rowing ergometer as well as a semi-recumbent cycle ergometer. The test protocol included a pre-exercise stage followed by incremental exercise until VO2max was reached. The order of exercise modes was randomized and there was a wash-out period between the two tests. Oxygen uptake was obtained via a breath-by-breath metabolic cart (Vmax™ Encore, San Diego, CA). Rowing VO2max was higher than cycling VO2max in both groups (p<0.05). Older subjects had less of an increase in VO2max from cycling to rowing (p<0.05). There was a significant relationship between muscle mass and VO2max for both groups (p<0.05). After correcting for muscle mass, the difference in cycling VO2max between groups disappeared (p>0.05), however, older subjects still demonstrated a lower rowing VO2max relative to younger subjects (p<0.05). Muscle mass is associated with the VO2max obtained, however, it appears that VO2max in older subjects may be less influenced by muscle mass than in younger subjects.


Prevention of cytomegalovirus disease in solid organ transplant patients: prophylactic versus preemptive therapy.
The advantages and disadvantages of universal prophylaxis and preemptive therapy and current evidence-based recommendations for preventing cytomegalovirus (CMV) disease in solid organ transplant recipients are discussed. Advantages of universal prophylaxis include the ease of implementation, a reduced incidence of CMV disease, and possibly fewer indirect effects of CMV infection. Disadvantages of universal prophylaxis may include prolonged antiviral drug exposure, resistance, toxicity, the development of late-onset CMV disease, and greater drug costs. Advantages of preemptive therapy may include reduced drug exposure and decreased risk for toxicity and resistance. Disadvantages include the logistic demands of laboratory testing, uncertainty about the impact on the indirect effects of CMV disease, and the costs associated with failure to prevent CMV disease. Evidence-based guidelines call for universal prophylaxis for patients at highest risk for CMV disease. Preemptive therapy may be most appropriate for those at a moderate or low risk of CMV. Antiviral drug regimens used for universal prophylaxis depend on the type of organ transplanted and the donor-recipient CMV serostatus. The optimal preemptive drug regimen and laboratory monitoring strategy are unknown. Selection of a strategy for preventing CMV disease in solid organ transplant patients requires consideration of patient-specific risk factors as well as practical considerations, such as available resources.


Effects of treatment for diabetes mellitus on circulating vascular progenitor cells.
The circulating endothelial progenitor cells (EPCs) have an important role in angiogenesis, and the smooth muscle progenitor cells (SMPCs) participate in atherosclerosis. However, little is known about the effects of treatment of diabetes mellitus (DM) on EPCs and SMPCs. Therefore, we investigated the relations between the number of circulating vasucular progenitor cells before and after the treatment for DM. Ten previously untreated DM patients were enrolled in this study. Blood samples were collected before and after treatment. The peripheral mononuclear cells were purified and cultured to differentiate them into EPCs and SMPCs. After two weeks, the number of EPCs was determined by Dil-labeled acetylated low density lipoprotein and lectin binding. The number of SMPCs was evaluated by immunocytochemical staining of alpha-smooth muscle actin. Before treatment, the number of EPCs and SMPCs was significantly related to hemoglobin A1c and blood sugar. Serial examination revealed that improvement of glycemic control significantly increased the number of both EPCs and SMPCs. DM reduces the number of circulating EPCs and SMPCs according to its severity, and treatment of DM significantly increases the number of EPCs and SMPCs, which may be involved in angiogenesis and atherosclerosis in diabetes.


Sequence-specific modification of mouse genomic DNA mediated by gene targeting techniques.
The major impact of the human genome sequence is the understanding of disease etiology with deduced therapy. The completion of this project has shifted the interest from the sequencing and identification of genes to the exploration of gene function, signalling the beginning of the post-genomic era. Contrasting with the spectacular progress in the identification of many morbid genes, today therapeutic progress is still lagging behind. The goal of all gene therapy protocols is to repair the precise genetic defect without additional modification of the genome. The main strategy has traditionally been focused on the introduction of an expression system designed to express a specific protein, defective in the transfected cell. But the numerous deficiencies associated with gene augmentation have resulted in the development of alternative approaches to treat inherited and acquired genetic disorders. Among these one is represented by gene repair based on homologous recombination (HR). Simply stated, the process involves targeting the mutation in situ for gene correction and for restoration of a normal gene function. Homologous recombination is an efficient means for genomic manipulation of prokaryotes, yeast and some lower eukaryotes. By contrast, in higher eukaryotes it is less efficient than in the prokaryotic system, with non-homologous recombination being 10-50 fold higher. However, recent advances in gene targeting and novel strategies have led to the suggestion that gene correction based on HR might be used as clinical therapy for genetic disease. This site-specific gene repair approach could represent an alternative gene therapy strategy in respect to those involving the use of retroviral or lentiviral vectors to introduce therapeutic genes and linked regulatory sequences into random sites within the target cell genome. In fact, gene therapy approaches involving addition of a gene by viral or nonviral vectors often give a short duration of gene expression and are difficult to target to specific populations of cells. The purpose of this paper is to review oligonucleotide-based gene targeting technologies and their applications on modifying the mouse genome.


Antiaromatase agents: evolving role in adjuvant therapy.
The goal of adjuvant hormonal therapy is to prevent breast cancer recurrence. Standard therapy with tamoxifen has shown great value in the adjuvant setting; however, its tolerability profile can render it unsuitable for some patients. The aromatase inactivator, exemestane, and the 2 aromatase inhibitors, letrozole and anastrozole, have been shown to be equivalent or superior to tamoxifen with respect to multiple endpoints in patients with metastatic breast cancer. With tolerability profiles that are similar to, and in many cases, more acceptable than that of tamoxifen, and efficacy potentially superior to tamoxifen, studies using the antiaromatase agents as adjuvant therapy are currently ongoing. These trials will answer some important questions, such as the order in which adjuvant hormonal therapies are selected to maximize efficacy, whether the antiaromatase agents show improved tolerability, and whether combination therapy is more effective than monotherapy.


Influence of the number and interval of treatment cycles on cytokine-induced killer cells and their adjuvant therapeutic effects in advanced non-small-cell lung cancer (NSCLC).
Cytokine-induced killer (CIK) cells have important therapeutic effects in adoptive cell transfer (ACT) for the treatment of various malignancies. In this study, we focused on in vitro expansion of CIK cells and their clinical efficacy in combination with chemotherapy in patients with advanced non-small-cell lung cancer (NSCLC). A total of 64 patients with NSCLC (enrolled from 2011 to 2012), including 32 patients who received chemotherapy alone or with sequential radiotherapy (conventional treatment, control group) and 32 patients who received conventional treatment and sequential CIK infusion (study group), were retrospectively analyzed. The time to progression (TTP), overall survival (OS) and adverse effects were analyzed and the phenotype of lymphocytes in CIK population was also determined by flow cytometry. After in vitro expansion, the average percentage of CIK cells was 26.35%. During the 54-month follow up, the median OS and TTP were significantly longer in the study group than in the control group (P=0.0189 and P=0.0129, respectively). The median OS of the ACT≥4cycles subgroup was significantly longer than that of the ACT<4cycles subgroup (P=0.0316). The percentage of CIK cells in patients who received ≥4cycles of ACT was higher than that in patients treated with <4cycles of ACT (P=0.0376). Notably, CIK cells were difficult to expand in vitro in some patients after the first ACT cycle but became much easier as the treatment cycles increased monthly. Longer treatment interval negatively impacted the expansion of CIK cells. Systematic immune levels can be increasingly boosted by reinfusion of ACT. Conventional treatment plus CIK cells is an effective therapeutic strategy to prevent progression and prolong survival of patients with advanced NSCLC.


[Transmembrane transport behavior of in vitro HepG2 cells of ananas and its effect on lipids and glucose distribution].
Pineapple (Ananas comosus) leaves contain mainly phenolic components with antioxidant and hypolipidemic effects. One of the principle components is p-coumaric acid. In this study, the transport behavior of p-coumaric acid, was observed after the administration of pineapple leaf phenols in vitro. Simultaneously, the effect of the phenols on glucose, total cholesterol and triglycerides transportation and metabolism in HepG2 cells was also observed. The results showed that the phenols had good transport characteristics. 5 min after the administration, p-coumaric acid of the phenols could be detected, and the content of p-coumaric acid reached the peak concentration after 60 min of the administration. p-coumaric acid of phenols have time-and dose-dependent manner. While promoting glucose transporter (GLUT4) and low density lipoprotein receptor (LDLR) expression, the phenols decreased intracellular lipid content. This reduction of intracellular lipid content was highly correlated with the promotion of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) expression, while the reduction of intracellular glucose levels was correlated with glycogen synthesis in the cells.


The effects of compound 48/80, morphine, and mast cell depletion on electroshock seizure in mice.
The effects of compound 48/80 (C48/80), morphine, and mast cell depletion on maximal electroshock seizure (MES) were studied in Swiss albino mice. An electrical current (60Hz, 0.2 msec) inducing convulsions in 50% of the animals (CC50) was assessed as 46 mA. Compound 48/80 (5 mg/kg) and morphine (100mg/kg) were administered subcutaneously. CC50 was applied separately to electroshock-unexposed animal groups at 15, 30, 60, 120, and 240 min after the onset of the experiment. In untreated controls, the percent of seizure induced by CC50 and percent of death among mice having convulsions were 50 and 20, respectively. After C48/80, a significant increase in rates of seizure at 60th and 120th min and death beyond 60th min (p < .0001) indicates a pro-convulsive action of the drug, probably caused by a reduction in MES threshold. In contrast, rate of seizure tended to decrease following mast-cell depletion, which was readily reversed by C48/80 at the 60th min (p < .0001). Mast-cell depletion, alone or plus morphine, significantly increased the death percentage of convulsions. Morphine alone reduced the percentage of seizure induced by the application of CC50 in the mast-cell depleted animals (anticonvulsive action) but increased the percent of dying animals by as much as 100% at the 30th and 60th min (p < .0001). Combined morphine + C48/80 not only augmented the anticonvulsive effect of morphine at the 30th min but also nullified the rate of death among mice having convulsions. We concluded that compound 48/80 (1) penetrates into the central nervous system to produce a central effect; (2) acts as pro-convulsive, and (3) paradoxically augments the anticonvulsive action of morphine, likely caused by the ability of the compound to increase the permeability of blood-brain barrier for morphine or by the release of histamine from mast cells in the brain, acting as anticonvulsant through the stimulation of H1 receptors or both. The precise mechanism of the increased death rate by C48/80 or morphine in intact and in mast-cell-depleted mice appears to involve pro-convulsive effects, cardiovascular impairment, and respiratory depression. The nullification of morphine-induced lethal toxicity by C48/80 could be due to the antagonistic interaction of the drug with opiate receptors in the brain.