Investigating the Potential of Isolating and Expanding Tumour-Infiltrating Lymphocytes from Adult Sarcoma.
Sarcomas are a heterogeneous group of mesenchymal neoplasms, many of which are associated with a high risk of metastasis and poor prognosis. Conventional chemotherapy and targeted therapies have varying effects across individuals and tumour subtypes. The current therapies frequently provide limited clinical benefit; hence, more effective treatments are urgently needed. Recent advances in immunotherapy, such as checkpoint inhibition or adoptive cell therapy (ACT), show potential in increasing efficacy by providing a more personalized treatment. Therapy with tumour-infiltrating lymphocytes (TILs) is an emerging field in immunotherapy. Here, we collected 190 sarcoma tumour specimens from patients without pre-operative adjuvant treatment in order to isolate TILs. We compared different methods of TIL expansion and optimized a protocol specifically for efficacy in culturing TILs from sarcoma. The expanded TIL populations were characterized by flow cytometry analysis using CD3, CD4, CD8, CD14, CD19 and CD56 markers. The TIL populations were non-specifically stimulated to establish TIL reactivity. Through an optimized expansion protocol, TILs were isolated and cultured from 54 of 92 primary sarcoma specimens. The isolated TILs varied in CD4+ and CD8+ T-cell compositions and retained their ability to release IFNγ upon stimulation. Our results suggest that certain sarcoma subtypes have the potential to yield a sufficient number of TILs for TIL therapy.


Length of intensive care unit stay and the apparent efficacy of antimicrobial-based versus non-antimicrobial-based ventilator pneumonia prevention interventions within the Cochrane review database.
The risk of acquiring ventilator-associated pneumonia (VAP) increases with intensive care unit (ICU) length of stay (LOS). The objectives here are to estimate, using data derived from randomized concurrent control trials (RCCTs) of non-antimicrobial versus antimicrobial interventions, the relation of LOS with firstly, apparent VAP prevention effect, and secondly, with VAP incidence in control and intervention groups. Control and intervention group data derived from 13 Cochrane reviews of 78 RCCTs of antimicrobial-based interventions versus 111 RCCTs of various non-antimicrobial-based VAP prevention interventions. In meta-regression models of VAP prevention effect versus group mean LOS, the effect size of non-antimicrobial-based interventions regress towards the null (+0.028; +0.002 to +0.054) whereas antimicrobial-based interventions regress away from the null (-0.043; -0.08 to -0.004). The day 9-10 VAP incidence increase is 1.28 (0.97-1.6) percentage points among the control groups of antimicrobial interventions per day. By contrast, these increases among antimicrobial- (0.45; 0.19-0.71) and non-antimicrobial- (0.58; 0.29-0.87) intervention groups and in control groups of non-antimicrobial- (0.76; 0.46-1.05) interventions are all similar. Antimicrobial-based versus non-antimicrobial-based interventions show overall greater apparent VAP prevention which is most apparent with longer group mean LOS. The basis for this surprising relationship with LOS resides, paradoxically, within the control rather than the intervention groups. This discrepancy implicates indirect (spill-over) effects, inapparent within individual antimicrobial-based RCCTs, which could spuriously conflate the appearance of VAP prevention.


Bowing fracture of the inferior angle of the scapula, a difficult diagnosis.
A 4-year-old boy presented with swelling over the inferior tip of the scapula and an unclear history. Initial radiographic findings were concerning for an aggressive lesion. This case highlights how a multimodality imaging approach was used to relieve uncertainty by diagnosing a paediatric bowing type fracture of the scapular tip.


Energy balance in cancer survivors at risk of weight gain: a review.
The study of energy balance [i.e., energy intake (EI) and energy expenditure (EE)] is a powerful tool for understanding body weight regulation and may contribute to our understanding of rapid weight gain risk in certain cancer survivors post-diagnosis. The purpose of this review was to summarize studies that assessed longitudinal, prospective changes in components of energy balance from diagnosis/start of treatment to any duration of follow-up in cancer survivors with prior evidence of weight gain (breast, prostate, thyroid, gynecologic, testicular, and acute lymphoblastic leukemia) RESULTS: The available literature suggests that energy balance components may be altered in cancer survivors who have a heightened risk of weight gain post-diagnosis. The evidence for EI was overall inconsistent. Conversely, decreases in resting and physical activity EE during the active phases of treatment (e.g., chemotherapy, hypothyroid state) were commonly noted, which then slowly rebounded towards baseline levels at the end of treatment and during follow-up assessments. Much of this evidence is based on data collected from breast cancer survivors, which highlights a paucity of data currently available on other cancer types. While there is growing acknowledgement that weight management interventions in cancer survivors are needed, it is important to recognize that changes in both behavioral (EI, physical activity EE) and passive (resting EE, thermic effect of food) components of energy balance may occur post-diagnosis. This information can help to inform weight management interventions which often entail modifications in diet and/or physical activity.


Inhibition of mutant troponin C activity by an intra-domain disulphide bond.
Triggering of contraction in striated muscles involves a conformational transition in the N-terminal domain of troponin C, the calcium-binding component of thin filaments. We have designed a mutant troponin C in which the key conformational transition and the calcium-regulatory activity are reversibly blocked by the formation of a disulphide bridge. Our results may be applicable to other proteins of the same family of calcium-binding proteins.


Stress as a neuroinflammatory condition in brain: damaging and protective mechanisms.
Several neuropsychiatric diseases are related with stress (posttraumatic stress disorder, major depressive disorder, anxiety disorders, schizophrenia) and stress exposure modifies the onset and evolution of some neurological diseases (neurodegenerative diseases). It is accepted that brain inflammatory responses contribute to cell damage during these illnesses. Studies carried out with some stress protocols (physical, psychological or mixed) show a pro-inflammatory response in the brain and other systems mainly characterized by a complex release of several inflammatory mediators such as cytokines, prostanoids, free radicals and transcription factors. This review considers the current status of knowledge of stress-induced inflammation in the brain. Interestingly, anti-inflammatory pathways are also activated in brain in response to stress, constituting a possible endogenous mechanism of defence against excessive inflammation. The possibility of pharmacological modulation of these pathways to prevent the accumulation of pro-inflammatory mediators and subsequent brain damage in stress and in stress-related neuropsychological conditions is also reviewed. This dual response elicited by stress in brain, both pro- and anti-inflammatory deserves further attention in order to understand pathophysiological changes as well as possible new therapeutic approaches of stress-related neuropsychopathologies.


Plasma brain natriuretic peptide at rest and after adenosine-induced myocardial ischemia in normotensive and essential hypertensive patients with suspected coronary artery disease.
This study investigated plasma brain natriuretic peptide (BNP) levels in normotensive and hypertensive patients with suspected coronary artery disease during radionuclide pharmacological stress testing. Twenty-seven normotensive patients (15 males, aged 63.0+/-4.5 years and 12 females, aged 63.0+/-4.1 years) and 38 essential hypertensive patients (25 males, aged 63.3+/-3.3 years and 13 females, aged 64.6+/-2.6 years) with chest pain and exercise stress testing inconclusive for coronary artery disease underwent myocardial perfusion single-photon emission computed tomography (SPECT) using adenosine infusion. SPECT identified patients without (16 normotensive and 22 hypertensive) and patients with (11 normotensive and 16 hypertensive) transient perfusion defects. Basal BNP levels in normotensive patients without transient myocardial ischemia (3.1+/-1.2 fmol/ml) were significantly (P<0.01) lower than those observed in normotensive patients with transient ischemia (8.2+/-1.2 fmol/ml), whereas BNP levels in hypertensive patients without transient ischemia (8.2+/-1.0 fmol/ml) did not significantly differ from those in hypertensive patients with transient ischemia (8.1+/-2.0 fmol/ml). No significant difference was found in BNP levels between males or females either in normotensive or hypertensive patients without or with ischemia. Adenosine infusion did not significantly change BNP levels in any subject group without or with myocardial perfusion defects. Our findings show that increases in BNP allow early detection of myocardial ischemia in normotensive patients, but not in hypertensive patients with suspected coronary artery disease. Adenosine-induced myocardial ischemia does not affect BNP production already activated by coronary artery disease in normotensive patients and by hemodynamic changes in hypertensive patients.


Apoptosis signaling by the novel compound 3-Cl-AHPC involves increased EGFR proteolysis and accompanying decreased phosphatidylinositol 3-kinase and AKT kinase activities.
The threonine and serine protein kinase AKT plays a major role in inhibiting apoptosis in a number of malignant cell types including prostate and breast carcinoma. Activation of AKT is a complex process involving translocation to the plasma membrane and phosphorylation of serine and threonine amino-acid residues. We now report that the novel compound 4-[3-(1-adamantyl)-4-hydroxyphenyl]-3-chlorocinnamic acid (3-Cl-AHPC), induces apoptosis in breast and prostate carcinoma cells and inhibits AKT activity in these cells. Overexpression of a constitutively activated AKT inhibits 3-Cl-AHPC-mediated apoptosis. Decrease in AKT activity occurs through 3-Cl-AHPC inhibition of phosphatidylinositol 3 kinase (PI3-K) activity. 3-Cl-AHPC inhibits PI3-K activity by enhancing epidermal growth factor receptor (EGFR) proteolysis and thus inhibiting EGFR association with the p85 subunit of PI3-K. 3-Cl-AHPC-mediated decrease in PI3-K activity results in the reduced synthesis of phosphatidylinositol 3,4 bisphosphate and phosphatidylinositol 3,4,5 triphosphate with the subsequent inhibition of integrin-linked kinase activity and serine-473 phosphorylation of AKT. Overexpression of EGFR results in increased AKT activity and inhibition of 3-Cl-AHPC-mediated decrease in AKT activation, AKT activity and 3-Cl-AHPC-mediated apoptosis. Inhibition of AKT activity by this compound results in the inability of AKT to phosphorylate and inactivate the proapoptotic forkhead transcription factor.


Regulation of cell growth by IRF-1 in BHK-21 cells.
Most cell lines that are used for the production of recombinant proteins proliferate spontaneously at a high rate. In many types of cultivation systems these cells still keep growing after having reached the desired cell density. Further proliferation in batch cultures leads to cell death as a consequence of nutrient and oxygen depletion as well as to accumulation of lactate and toxic products. Consequently, in many technical processes, the surplus of cells is removed.We have established cell lines in which proliferation is controlled by a physiological regulator, IRF-1. IRF-1 (Interferon Regulatory Factor 1) is a transcriptional activator and acts as a tumor suppressor. Constitutive overexpression of recombinant IRF-1 leads to inhibition of cell growth. The extent of this growth arrest depends on the intracellular concentration of active IRF-1. To allow IRF-1 expression in various mammalian cells a system for conditional IRF-1 activation has been established. A fusion protein composed of IRF-1 and the hormone binding domain of the human estrogen receptor, was used. This system allows to control gradually the growth of several mammalian cell lines by adjusting the intracellular concentration of active IRF-1 via estradiol in the medium. We have evaluated BHK-21 cells with respect to IRF-1 mediated cell growth inhibition and expression of two secreted proteins. Whereas the productivity of proliferation inhibited cells with respect to constitutively transcribed IgG genes is reduced, productivity of another secreted protein which is controlled by an IRF-1 inducible promoter is strongly enhanced under these conditions.


Changes in phospholipid composition, protein content and chemical properties of flaxseed oil during development.
The aim of the present research is to investigate the effect of harvest date on the composition of flaxseed. Samples were collected at regular intervals from 7 to 56 days after flowering (DAF) and analyzed for phospholipid composition, storage protein content and chemical properties. Phospholipid (PL) percentage of the total lipid decreased from 32.72% on the 7th DAF to 2.55% on the 56th DAF. The most phospholipids present in flaxseed were phosphatidylinositol (PI), phosphatidylethanolamine (PE) and lysophosphatidylcholine (LPC) which were highly unsaturated and rich in linolenic and linoleic acids, comprising together 60% of the total fatty acids. Chemical investigation of flaxseed oil showed overall a decrease in UV absorbance (K(232) and K(270)), acid value, free fatty acid content and an increase in peroxide value and storage protein content with development. At full maturity, flaxseed contained 29% proteins on a dry weight basis (DW %).


Nicotine-induced NO-mediated increase in cortical cerebral blood flow is blocked by beta2-adrenoceptor antagonists in the anesthetized rats.
Involvement of nitric oxide (NO) and beta-adrenoceptors in an increase in the cortical cerebral blood flow (CBF) following an intravenous (i.v.) injection of a small dose of nicotine which did not affect the systemic blood pressure in the rats was investigated. I.v. injection of nicotine (30 microg/kg) for 1 min produced a significant increase in CBF lasting for more than 20 min without a significant effect on the systemic blood pressure. I.v. injection of L-N(G)-nitroarginine methylester (30 mg/kg) significantly attenuated nicotine-induced increase in the cortical CBF. The attenuation was reversed by i.v. injection of L-arginine (300 mg/kg), suggesting an intimate role of nitric oxide (NO) in nicotine-induced increase in the cortical CBF. The nicotine-induced increase in the cortical CBF was significantly attenuated by propranolol (10 mg/kg, i.v.) and ICI 118,551 (a beta2-adrenoceptor antagonist, 10 mg/kg, i.v.) but not by metoprolol (a beta1-adrenoceptor antagonist, 10 mg/kg, i.v.). Beta2-adrenoceptors on presynaptic nitrergic nerves may be involved in nicotine-induced NO-mediated increase in the cortical CBF.


Quantum-confinement and Structural Anisotropy result in Electrically-Tunable Dirac Cone in Few-layer Black Phosphorous.
Two-dimensional (2D) materials are well-known to exhibit interesting phenomena due to quantum confinement. Here, we show that quantum confinement, together with structural anisotropy, result in an electric-field-tunable Dirac cone in 2D black phosphorus. Using density functional theory calculations, we find that an electric field, E ext, applied normal to a 2D black phosphorus thin film, can reduce the direct band gap of few-layer black phosphorus, resulting in an insulator-to-metal transition at a critical field, Ec. Increasing E ext beyond Ec can induce a Dirac cone in the system, provided the black phosphorus film is sufficiently thin. The electric field strength can tune the position of the Dirac cone and the Dirac-Fermi velocities, the latter being similar in magnitude to that in graphene. We show that the Dirac cone arises from an anisotropic interaction term between the frontier orbitals that are spatially separated due to the applied field, on different halves of the 2D slab. When this interaction term becomes vanishingly small for thicker films, the Dirac cone can no longer be induced. Spin-orbit coupling can gap out the Dirac cone at certain electric fields; however, a further increase in field strength reduces the spin-orbit-induced gap, eventually resulting in a topological-insulator-to-Dirac-semimetal transition.


Prevalence of sleep apnea in HRH Princess Maha Chakri Srinthorn Medical Center, Thailand.
Many studies have been conducted to find the prevalence of obstructive sleep apnea (OSA) in Western countries. Limited data have been demonstrated in Asian countries; however, no such data have been reported in Thailand. The purpose of this study was to examine the prevalence of OSA and their related factors in Thai adults. This study was performed in a university hospital in a rural area of central Thailand and data were collected. The population included office-based workers and people who came to contact with the hospital. Among the total of 2,685 participants at the baseline examination, a random sample of 199 men and women was studied with employment of overnight full polysomnography to determine the prevalence of OSA and OSA syndrome (OSAS). The percentage of habitual snorers in the study population was 26.4%. The prevalence of OSA (apnea-hypopnea index 5) was 11.4%. When OSAS was defined by an apnea-hypopnea index 5 plus excessive daytime sleepiness, its prevalence was 4.4%. The prevalence of OSA and OSAS in men was 15.4% and 4.8% and in women was 6.3% and 1.9%, respectively. Logistic regression analyses showed that gender, body mass index, age, hypertension, alcohol consumption, and smoking were associated with the risk of OSA. This study demonstrates that OSA is a common problem in the Thai adult population as in Western and other Asian countries. Understanding and treatment may be essential to reduce the risk of related medical problems.


Use and understanding of the nutrition information panel of pre-packaged foods in a sample of Mexican consumers.
To assess the use and understanding of the Nutritional information Panel (NIP) of pre-packaged foods by Mexican consumers. A questionnaire and an understanding test for NIP were applied to adult consumers in supermarkets of six cities in the Northern, Central, and Southern regions of Mexico. Data were analyzed by frequencies and Poisson regression models. Interviewed 731 consumers; 71.5% were women, mean age 33 ± 9.7 (range: 18-60), 70% completed high-school or a higher degree. In total, 17% of consumers use the NIP for making purchase decisions; 49% did not understand the NIP. Only 1.2% of consumers answered correctly the five questions of the NIP understanding test. The use and understanding of the NIP are low despite a high proportion self-reported reading and understanding. The lack of previous knowledge of the technical language prevents use and interpretation of NIP nutritional information for purchasing decisions.


The activity of the Drosophila Vestigial protein is modified by Scalloped-dependent phosphorylation.
The Drosophila vestigial gene is required for proliferation and differentiation of the adult wing and for differentiation of larval and adult muscle identity. Vestigial is part of a multi-protein transcription factor complex, which includes Scalloped, a TEAD-class DNA binding protein. Binding Scalloped is necessary for translocation of Vestigial into the nucleus. We show that Vestigial is extensively post-translationally modified and at least one of these modifications is required for proper function during development. We have shown that there is p38-dependent phosphorylation of Serine 215 in the carboxyl-terminal region of Vestigial. Phosphorylation of Serine 215 occurs in the nucleus and requires the presence of Scalloped. Comparison of a phosphomimetic and non-phosphorylatable mutant forms of Vestigial shows differences in the ability to rescue the wing and muscle phenotypes associated with a null vestigial allele.


Age and dementia effect on neuropsychological test performance in very old age--influence of risk factors for dementia.
In old age a large part of the variance in cognitive performance in population samples is explained by normal aging; in addition many subjects over 80 years are demented and therefore dementia also explains a part of cognitive variability. The question is whether the different factors for dementia (such as ApoE4, external atrophy parameter of the cranial computer tomography [cCT], education, sex or serum zinc level) influence the relation between age or dementia and Mini Mental State (MMSE) performance. In an epidemiological study data were analyzed of N = 239 subjects for the above factors. Most statistically significant variables of the MMSE do not change the amount of the partial correlation coefficient between the parameters age or dementia and MMSE. The external atrophy, however, diminishes the magnitude of the partial correlation between age and MMSE. In contrast the dementia-MMSE relation is unchanged. This points to a generally similar factor structure of cognitive aging and dementia in old age, but differences exist with respect to the importance of the external atrophy parameter of the brain. Most factors investigated explain separate parts of variance of cognitive performance in old age.


Mitotic centromere-associated kinesin (MCAK) mediates paclitaxel resistance.
Paclitaxel has powerful anticancer activity, but some tumors are inherently resistant to the drug, whereas others are initially sensitive but acquire resistance during treatment. To deal with this problem, it will be necessary to understand the mechanisms of drug action and resistance. Recent studies indicate that paclitaxel blocks cell division by inhibiting the detachment of microtubules from centrosomes. Here, we demonstrate that mitotic centromere-associated kinesin (MCAK), a kinesin-related protein that destabilizes microtubules, plays an important role in microtubule detachment. Depletion of MCAK altered mitotic spindle morphology, increased the frequency of lagging chromosomes, and inhibited the proliferation of WT CHO cells, confirming that it is an essential protein for cell division. In contrast, MCAK depletion rescued the proliferation of mutant paclitaxel-dependent cell lines that are unable to divide because of defective spindle function resulting from altered α-tubulin or class III β-tubulin overexpression. In concert with the correction of mitotic defects, loss of MCAK reversed an aberrantly high frequency of microtubule detachment in the mutant cells and increased their sensitivity to paclitaxel. The results indicate that MCAK affects cell sensitivity to mitotic inhibitors by modulating the frequency of microtubule detachment, and they demonstrate that changes in a microtubule-interacting protein can reverse the effects of mutant tubulin expression.


Metabolism and action of amino acid analog anti-cancer agents.
The preclinical pharmacology, antitumor activity and toxicity of seven of the more important amino acid analogs, with antineoplastic activity, is discussed in this review. Three of these compounds are antagonists of L-glutamine: acivicin, DON and azaserine; and two are analogs of L-aspartic acid: PALA and L-alanosine. All five of these antimetabolites interrupt cellular nucleotide synthesis and thereby halt the formation of DNA and/or RNA in the tumor cell. The remaining two compounds, buthionine sulfoximine and difluoromethylornithine, are inhibitors of glutathione and polyamine synthesis, respectively, with limited intrinsic antitumor activity; however, because of their powerful biochemical actions and their low systemic toxicities, they are being evaluated as chemotherapeutic adjuncts to or modulators of other more toxic antineoplastic agents.


Accelerating the granulation of anammox sludge in wastewater treatment with the drive of "micro-nuclei": A review.
Anammox granule sludge (AnGS) has great potential in the field of wastewater nitrogen removal, but its development and promotion have been limited by the slow granulation speed and fragile operating stability. Based on the reviews about the AnGS formation mechanism in this paper, "micro-nuclei" was found to play an important role in the granulation of AnGS, and adding "micro-nuclei" directly into the reactor may be an efficient way to accelerate the formation of AnGS. Then, accelerating AnGS granulation with inert particles, multivalent positive ions, and broken granule sludge as "micro-nuclei" was summarized and discussed. Among inert particles, iron-based particles may be a more advantageous candidate for "micro-nuclei" due to their ability to provide attachment sites and release ferric/ferrous ions. The precipitations of multivalent positive ions are also a potential option for "micro-nuclei" that can be generated in-situ, but a suitable dosing strategy is necessary. About broken granular sludge, the broken active AnGS may have advantages in terms of anaerobic ammonium oxidation bacteria-affinity and granulation speed, while using inactive granular sludge as "micro-nuclei" can avoid interfering bacterial invasion and has a higher cost performance than broken active AnGS. In addition, possible research directions for accelerating the formation of AnGS by dosing "micro-nuclei" were highlighted. This paper is intended to provide a possible pathway for the rapid start-up of AnGS systems, and references for the optimization and promotion of the AnGS process.


[Biliary atresia].
Biliary atresia is a rare, neonatal, progressive cholestatic liver disease caused by fibroinflammatory obstruction of the biliary tree. Typical symptoms include prolonged neonatal jaundice, pale stools, and conjugated hyperbilirubinemia. Treatment starts with portoenterostomy, where fibrotic remnants of the extra hepatic bile ducts are replaced with small intestine. Best operative results are achieved among the youngest patients treated in specialized centres. Despite clearance of jaundice after a successful operation, fibrotic change of the liver continues in most. Liver transplantation serves as a salvage procedure if the portoenterostomy fails, or complications of liver cirrhosis develop after an initially successful portoenterostomy. In Finland, biliary atresia treatment was centralized in 2005. Of the patients treated thereafter, 90% are alive and over 80% with their native livers.


A new and efficient synthesis of 4-functionalized benzo[b]furans from 2,3-dihalophenols.
Tandem Sonogashira coupling/5-endo-dig cyclization reactions on 2,3-dihalophenols suppose a straightforward entry to 4-halobenzo[b]furans, which can be easily transformed into 4-functionalized benzo[b]furans, that are difficult to synthesize by other procedures. On the other hand, the starting 2,3-dihalophenols are efficiently prepared from commercially available 3-halophenols, via their N,N-diethyl carbamates by selective lithiation at the 2-positions by treatment with s-BuLi/TMEDA or LDA at low temperature and reaction with halogen electrophilic reagents.


Interaction of DNA-lesions induced by sodium fluoride and radiation and its influence in apoptotic induction in cancer cell lines.
Fluoride is an essential trace element but also an environmental contaminant with major sources of exposure being drinking water, food and pesticides. Previous studies showed that sodium fluoride (NaF) at 5 mM or more is required to induce apoptosis and chromosome aberrations and proposed that DNA damage and apoptosis play an important role in toxicity of excessive fluoride. The aim of this study is directed to understand the nature of DNA-lesions induced by NaF by allowing its interaction with radiation induced DNA-lesions. NaF 5 mM was used after observing inability to induce DNA damages and apoptosis by single exposure with 50 μM or 1 mM NaF. Co-exposure to NaF and radiation significantly increased the frequency of aberrant metaphases and exchange aberrations in human lymphocytes and arrested the cells in G1 stage instead of apoptotic death. Flow cytometric analysis, DNA fragmentation and PARP-cleavage analysis clearly indicated that 5 mM NaF together with radiation (1 Gy) induced apoptosis in both U87 and K562 cells due to down regulation of expression of anti-apoptotic proteins, like Bcl2 in U87 and inhibitors of apoptotic proteins like survivin and cIAP in K562 cells. This study herein suggested that single exposure with extremely low concentration of NaF unable to induce DNA lesions whereas higher concentration induced DNA lesions interact with the radiation-induced DNA lesions. Both are probably repaired rapidly thus showed increased interactive effect. Coexposure to NaF and radiation induces more apoptosis in cancer cell lines which could be due to increased exchange aberrations through lesions interaction and downregulating anti-apoptotic genes.


Evaluation of Rifampin's Transporter Inhibitory and CYP3A Inductive Effects on the Pharmacokinetics of Venetoclax, a BCL-2 Inhibitor: Results of a Single- and Multiple-Dose Study.
Venetoclax is a selective, potent, first-in-class B-cell lymphoma-2 inhibitor that has demonstrated clinical efficacy in a variety of hematological malignancies. A single-dose and multiple-dose rifampin study was conducted to evaluate the effect of CYP3A induction and transporter inhibition on the pharmacokinetics of venetoclax. Subjects received a single dose of venetoclax 200 mg on day 1 of period 1 and days 1 and 14 of period 2, a single dose of rifampin 600 mg on day 1 of period 2, and rifampin 600 mg once daily on days 5 through 17 of period 2. Blood samples were collected up to 96 hours after each venetoclax dose on day 1 of period 1 and days 1 and 14 of period 2. Compared with venetoclax alone, coadministration with a single dose of rifampin increased venetoclax Cmax and AUC∞ by 106% (90%CI, 73%-145%) and 78% (90%CI, 50%-111%), respectively, whereas coadministration with multiple doses of rifampin decreased venetoclax Cmax and AUC∞ by 42% (90%CI, 31%-52%) and 71% (90%CI, 66%-76%), respectively. It was possible to isolate the net effect of chronic CYP3A induction from acute P-glycoprotein (P-gp) inhibition by comparing venetoclax exposures following coadministration with multiple doses of rifampin versus a single dose of rifampin, which showed that CYP3A induction decreased venetoclax Cmax and AUC by 72% and 84%, respectively. These results are consistent with venetoclax being a P-gp substrate and indicate that CYP3A plays a major role in venetoclax metabolism. Prescribers should consider agents with little or no CYP3A induction during treatment with venetoclax.


A dominant mutation in the stereocilia-expressing gene TBC1D24 is a probable cause for nonsyndromic hearing impairment.
Mutations in TBC1D24 have been linked to a variety of epileptic syndromes and recently to syndromic hearing impairment DOORS syndrome and nonsyndromic hearing impairment DFNB86. All TBC1D24 mutations reported so far were inherited in the recessive mode. In a dominant family segregated with late-onset, progressive, nonsyndromic hearing impairment, linkage analysis revealed a 2.07 Mb candidate region on chromosome 16p13.3 that contains TBC1D24. Whole-exome sequencing identified a heterozygous p.Ser178Leu variant of TBC1D24 as the only candidate mutation segregating with the hearing loss within the family. In perinatal mouse cochlea, we detected a restricted expression of Tbc1d24 in the stereocilia of the hair cells as well as in the spiral ganglion neurons. Our study suggested that the p.Ser178Leu mutation of TBC1D24 is a probable cause for dominant, nonsyndromic hearing impairment. Identification of TBC1D24 as the stereocilia-expressing gene may shed new light on its specific function in the inner ear.


Alcohol-soluble Star-shaped Oligofluorenes as Interlayer for High Performance Polymer Solar Cells.
Two star-shaped oligofluorenes with hexakis(fluoren-2-yl)benzene as core are designed and sythesized for interfacial materials in polymer solar cell. Diethanolamino groups are attached to the side chain of fluorene units for T0-OH and T1-OH to enable the alcohol solubility, and additional hydrophobic n-hexyl chains are also grafted on the increased fluorene arms for T1-OH. In conventional device with PCDTBT/PC71BM as active layer, a 50% enhanced PCE is obtained by incorporating T0-OH and T1-OH as the interlayer compared with device without interlayer. By optimizing the active material with PTB7 and with the inverted device structure, a maximum PCE of 9.30% is achieved, which is among the highest efficiencies for PTB7 based polymer solar cells. The work function of modified electrode, the surface morphology and the suraface properties are systematically studied. By modifying the structures of the star-shaped molecules, a balance between the hydrophobic and hydrophilic property is finely tuned, and thus facilitate the interlayer for high performance of PSCs.


Traffic, transport, and vegetation drive VOC concentrations in a major urban area in Texas.
The population of Texas has increased rapidly in the past decade. The San Antonio Field Study (SAFS) was designed to investigate ozone (O3) production and precursors in this rapidly changing, sprawling metropolitan area. There are still many questions regarding the sources and chemistry of volatile organic compounds (VOCs) in urban areas like San Antonio which are affected by a complex mixture of industry, traffic, biogenic sources and transported pollutants. The goal of the SAFS campaign in May 2017 was to measure inorganic trace gases, VOCs, methane (CH4), and ethane (C2H6). The SAFS field design included two sites to better assess air quality across the metro area: an urban site (Traveler's World; TW) and a downwind/suburban site (University of Texas at San Antonio; UTSA). The results indicated that acetone (2.52 ± 1.17 and 2.39 ± 1.27 ppbv), acetaldehyde (1.45 ± 1.02 and 0.93 ± 0.45 ppbv) and isoprene (0.64 ± 0.49 and 1.21 ± 0.85 ppbv; TW and UTSA, respectively) were the VOCs with the highest concentrations. Additionally, positive matrix factorization showed three dominant factors of VOC emissions: biogenic, aged urban mixed source, and acetone. Methyl vinyl ketone and methacrolein (MVK + MACR) exhibited contributions from both secondary photooxidation of isoprene and direct emissions from traffic. The C2H6:CH4 demonstrated potential influence of oil and gas activities in San Antonio. Moreover, the high O3 days during the campaign were in the NOx-limited O3 formation regime and were preceded by evening peaks in select VOCs, NOx and CO. Overall, quantification of the concentration and trends of VOCs and trace gases in a major city in Texas offers vital information for general air quality management and supports strategies for reducing O3 pollution. The SAFS campaign VOC results will also add to the growing body of literature on urban sources and concentrations of VOCs in major urban areas.


Association between lack of angiogenic response in muscle tissue and high expression of angiostatic ELR-negative CXC chemokines in patients with juvenile dermatomyositis: possible link to vasculopathy.
To investigate the relationship between the vasculopathy of juvenile dermatomyositis (juvenile DM) and the balance between the angiostatic ELR- and angiogenic ELR+ CXC chemokines in the muscle of patients with the disease. The expression of 3 ELR- CXC chemokines (interferon-inducible protein 10 [IP-10], monokine induced by interferon-gamma, and interferon-inducible T cell alpha-chemoattractant) and 2 ELR+ CXC chemokines was quantitated in muscle biopsy samples from 7 patients with juvenile DM and 7 healthy children, by real-time polymerase chain reaction. The findings were correlated with various histopathologic features, with particular emphasis on the degree of vasculopathy. Synovial biopsy specimens from patients with juvenile rheumatoid arthritis (JRA) were used for additional comparison. The angiostatic ELR- chemokines were expressed at high levels, while the angiogenic ELR+ chemokines were barely detectable, in most juvenile DM samples. This contrasted sharply with the findings in both normal muscle biopsy specimens and JRA synovial tissue specimens. The expression of the ELR- chemokines in juvenile DM samples correlated with the intensity of mononuclear cell infiltration. Furthermore, the juvenile DM samples with the highest degree of capillary loss had the highest levels of ELR- CXC chemokines. The presence of IP-10 in juvenile DM muscle specimens was confirmed by immunohistochemistry analysis. In addition, immunohistochemical staining of muscle tissue revealed that CXCR3, a receptor utilized by ELR- CXC chemokines, was expressed in vascular endothelial cells. Increased expression of the interferon-induced angiostatic ELR- CXC chemokines is a feature of juvenile DM that parallels the degree of vasculopathy in patients with the disease. Collectively, these findings are consistent with a model in which a subset of inflammatory cells secrete angiostatic ligands, which then contribute to a local atrophying effect on the muscle's vasculature via a receptor-mediated process.


Chronic Alcohol Intoxication Is Not Accompanied by an Increase in Calpain Proteolytic Activity in Cardiac Muscle of Rats.
Enzymatic activity of Ca2+-dependent calpain proteases as well as the content and gene expression of μ-calpain (activated by micromolar calcium ion concentrations), calpastatin (inhibitor of calpains), and titin (substrate for calpains) were investigated in cardiac muscles of rats subjected to chronic alcoholization for 3 and 6 months. There was no increase in the "heart weight/body weight" parameter indicating development of heart hypertrophy in the alcoholized rats, while a decreasing trend was observed for this parameter in the rats after 6-month modeling of alcoholic cardiomyopathy, which indicated development of atrophic changes in the myocardium. Fluorometric measurements conducted using the Calpain Activity Assay Kit did not reveal any changes in total calpain activity in protein extracts of cardiac muscles of the rats alcoholized for 3 and 6 months. Western blot analysis did not show reliable changes in the contents of μ-calpain and calpastatin, and SDS-PAGE did not reveal any decrease in the titin content in the myocardium of rats after the chronic alcohol intoxication. Autolysis of μ-calpain was also not verified, which could indicate that proteolytic activity of this enzyme in myocardium of chronically alcoholized rats is not enhanced. Using Pro-Q Diamond staining, changes in phosphorylation level of titin were not detected in cardiac muscle of rats after chronic alcoholization during three and six months. A decrease in µ-calpain and calpastatin mRNA content (~1.3-fold, p ≤ 0.01 and ~1.9-fold, p ≤ 0.01, respectively) in the myocardium of rats alcoholized for 3 months and decrease in calpastatin mRNA (~1.4-fold, p ≤ 0.01) in animals alcoholized for 6 months was demonstrated using real-time PCR. These results indicate negative effect of chronic alcohol intoxication on expression of the abovementioned genes.


Sub-lethal effects of cadmium and copper on RNA/DNA ratio and energy reserves in the green-lipped mussel Perna viridis.
This study aims to test if RNA/DNA ratio and various energy reserve parameters (i.e., glycogen, lipid, protein content and total energy reserves) are sensitive biomarkers for indicating stresses induced by metal contaminants in the green-lipped mussel Perna viridis, a common organism for biomonitoring in Southeast Asia. This study was, therefore, designed to examine the effects of cadmium (Cd) and copper (Cu) on these potential biomarkers in two major energy storage tissues, adductor muscle (AM) and hepatopancreas (HP), of P. viridis after sub-lethal exposure to either metal for 10 days. The results showed that neither Cd nor Cu treatments affected the RNA/DNA ratio, glycogen and protein contents in AM and HP. As the most sensitive biomarker in P. viridis, the total lipid content in both AM and HP was significantly decreased in the treatment of 5μg Cu/L and 0.01-0.1μgCd/L, respectively. However, soft-tissue body burdens of Cu or Cd did not significantly correlate with each of the four biomarkers regardless of the tissue type. In addition, AM generally stored more glycogen than HP, whereas HP stored more lipids than AM. We proposed that multiple biomarkers may be employed as an integrated diagnostic tool for monitoring the health condition of the mussels.


Development of a multigenic plasmid vector for HCV DNA immunization.
HCV viral nucleocapsid protein (C), non-structural protein 3 (NS3) and the envelope glycoproteins E1 and E2 are candidate immune targets for developing anti-HCV DNA vaccine. Nevertheless, the immune response elicited by these antigens often appears weak and/or transient. Different approaches have been studied for enhancing and/or modulating the immune response of the DNA vaccine. On the basis of a prototype multigenic plasmid vector constituted of two different transcription cassettes (pRC100), we have developed a plasmid vector that allows the independent and simultaneous expression of murine IL2 and of an antigenic domain of the HCV NS3 C terminus (pRC112-HCV). The highly conserved NS3 region spans from nt 4403 to nt 4829 and contains two putative B and T epitopes. The development of this multigenic plasmid vector may combine the expression and local production of an immunomodulatory molecule (mIL2) together with the possibility of addressing the host immune response to the most immunogenic and conserved epitopes, specifically tailored in the plasmid vector.


[Adjuvants and additives in vaccines--medical relevance].
Side effects of vaccinations can have different causes. Substances admixed to vaccines may produce allergic or toxic reactions. The significance and importance of causal associations is discussed in this paper. A table is added listing the most important substances in vaccines, such as inactivating substances, preservatives, stabilizers, adjuvants, and residual substances derived from production processes. Among possible allergic reactions, type I reactions, as the most undesirable ones, should be avoided. In this respect, vaccines against yellow fever are the most important ones. With respect to antibiotics, it should be stressed that penicillin and cephalosporins are not contained in any of the vaccines. The significance of side effects caused by ethylmercury as a preservative (thiomersal) is extensively discussed in the literature. Allergy against this substance is common among the population, manifested as type IV reactions following superficial antigen administration. It has been shown that deep intramuscular injection of thiomersal-containing vaccines may be administered even to persons who are allergic to this substance without risk of side effects. Regarding toxic side effects after application of thiomersal, several studies have disproved a causal relation between thiomersal exposure and developmental disorders. Nevertheless, the general recommendation is to use thiomersal-free vaccines, unless no other preparations are available. In these cases risk of morbidity and mortality from the vaccine-preventable diseases outweigh by far any theoretical risk from ethylmercury.


[Making patients actors in their own care].
Enhanced recovery after surgery is a caregiving method focusing on the patient. It aims at ensuring faster recovery of the patient's functional abilities after surgery for a rapid return to the everyday environment and activities with total safety.


Basic fibroblast growth factor decreases elastin gene transcription in aortic smooth muscle cells.
The extracellular matrix (ECM) protein elastin plays an essential role in the cardiovascular system by imparting elasticity to blood vessel wall. In this study, we examined the effect of basic fibroblast growth factor (bFGF) on the expression of elastin in aortic smooth muscle cells (SMC) to gain insight into events associated with cardiovascular diseases. The results show that bFGF treatment of SMC causes a significant decrease in elastin mRNA and secreted tropoelastin levels. Nuclear run-on analyses demonstrate that the downregulation is due to a decrease in the level of elastin gene transcription. Transient transfections of SMC with wild-type and mutated elastin gene promoter/chloramphenicol acetyl transferase (CAT) constructs show that a previously identified activator protein-1-cAMP response element (AP1/CRE) (-564 to -558-bp) within the elastin promoter mediates the bFGF-dependent downregulation of elastin gene transcription in SMC. Addition of bFGF to SMC activates the extracellular signal-regulated kinases 1/2 (ERK1/2) resulting in their translocation into the nucleus and subsequent induction of Fra-1. The addition of PD-98059, an inhibitor of ERK1/2 kinase, abrogates the bFGF-dependent decrease of elastin mRNA in SMC. The described inhibitory effect of bFGF on elastin gene expression in SMC may significantly contribute to the inefficient repair of elastin in early stages of vascular wall injury.


Regiospecific synthesis and anti-human immunodeficiency virus activity of novel 5-substituted N-alkylcarbamoyl and N,N-dialkylcarbamoyl 1,2,3-triazole-TSAO analogues.
Several 5-N-alkyl and 5-N,N-dialkylcarbamoyl substituted analogues of the anti-human immunodeficiency virus (HIV) type 1 lead compound [1-[2',5'-bis-O-(tert-butyldimethylsilyl)-beta-D-ribofuranosyl]-5- (N,N-dimethylcarbamoyl)-1,2,3-triazole]-3'-spiro-5"-(4"-amino-1",2"- oxathiole-2",2"-dioxide) have been prepared and evaluated as inhibitors of HIV-1 replication. A new regiospecific synthetic procedure is described. The compounds were prepared by cycloaddition of the appropriate glycosylazide to 2-oxoalkylidentriphenyl-phosphoranes, followed by treatment with primary or secondary amines, to yield, exclusively, 5-substituted 1,2,3-triazole-TSAO analogues. Several 5-substituted 1,2,3-triazole-TSAO derivatives proved to be potent inhibitors of HIV-1 replication with higher antiviral selectivity than that of the parent TSAO prototype.


Testing values of crested porcupine habitats by experimental food patches.
We established depletable, artificial food patches in three habitats used by Indian crested porcupines (Hystrix indica) in a desert biome, and measured the number of food items remaining (i.e., "giving up density"=GUD) following nightly foraging bouts. Porcupines discriminated between resource types (peanuts vs. garbanzo beans), and exhibited clear habitat preferences in the face of uniform resource availability in time and space. Lowest GUD's (=lowest foraging costs) were in the habitat of densest cover, and during dark (little or no moon) nights. The results indicated a high sensitivity to predation risk. Crested porcupines behaved as expected of optimal foragers, and appear to be excellent subjects for further field experiments using the GUD approach.


Antibrain antibodies in children with autism and their unaffected siblings.
Serum autoantibodies to human brain, identified by ELISA and Western immunoblotting, were evaluated in 29 children with autism spectrum disorder (22 with autistic disorder), 9 non-autistic siblings and 13 controls. More autistic subjects than controls had bands at 100 kDa in caudate, putamen and prefrontal cortex (p<0.01) as well as larger peak heights of bands at 73 kDa in the cerebellum and cingulate gyrus. Both autistic disorder subjects and their matched non-autistic siblings had denser bands (peak height and/or area under the curve) at 73 kDa in the cerebellum and cingulate gyrus than did controls (p<0.01). Results suggest that children with autistic disorder and their siblings exhibit differences compared to controls in autoimmune reactivity to specific epitopes located in distinct brain regions.


The role of reconstructive surgery after soft tissue sarcoma resection.
Soft tissue sarcomas are rare malignant tumors with pejorative prognosis. They require a multidisciplinary approach in a specialized hospital belonging to the NetSarc network in France. In all cases treated with curative intent, the objective of excision surgery is to achieve wide, microscopically negative margins (R0 according to the UICC classification). When growing on a limb, sarcomas may threaten functionally relevant structures and even lead to amputation. Nowadays, when combined with radiation therapy, wide exeresis limb-sparing surgery is achievable in 90 to 95% of the cases, of which 25% will nevertheless require reconstructive surgery to preserve the limb, to limit postoperative complications and to manage possible sequelae. Progress in reparative surgery, particularly in microsurgery, has helped not only to improve limb salvage rates but also to create wider margins without altering oncologic goals of curative resection. After determining the range of resection, reconstructive surgery should be tailored to address the tissue defect. The converse is to be strongly discouraged. The extent of resection must not be compromised or reduced in order to facilitate reconstructive surgery. A plastic surgeon must master all the flap techniques, including microsurgery, while taking into account the impact of preoperative and postoperative radiation therapy on previously irradiated tissues or on wounds requiring adjuvant therapy. Recent developments, especially as regards perforator flaps, have helped to enhanced the quality of reconstruction procedure while reducing donor site morbidity. In our experience, perforator flaps are a workhorse in reconstructive surgeries subsequent to soft tissue sarcoma of the extremities. On a parallel track, lipofilling (otherwise known as fat grafting or fat transfer) has become the first-line treatment for patients with post-surgical functional or cosmetic sequalae. It is performed after long-term follow-up during disease-free survival. Strict clinical examination and MRI are mandatory prior to programming any local procedure. Usually, three to four sessions of fat grafting are needed to enhance local trophicity or the cosmetic aspect. Sequalae treatments are of great interest in terms of psychological as well as functional outcome.


A novel large deletion in the APC gene associated with Gardner syndrome in a Chinese family.
Gardner syndrome is a hereditary disease characterized by familial adenomatous polyposis (FAP), accompanied by soft tissue tumors. a Chinese FAP family was enrolled and followed-up for three years. a novel large germline fragment deletion (EX10_16DEL) of the adenomatous polyposis coli (APC) gene was identified by multiplex ligation-dependent probe amplification (MLPA). An unexpected abdominal tumor grew two years after a subtotal colectomy of the proband. The immunohistochemistry study of the abdominal tumor showed SMA(focal+), calponin(+), β-catenin(nucleus+) and CD34(focal+), CD117(-), which was consistent with a desmoid tumor. when a FAP related desmoid tumor appears, the possibility of Gardner syndrome should be considered. This is the first largest deletion of the APC gene in the Chinese population associated with Gardner syndrome.


Comparing the Effect of Implanted Peroneal Nerve Stimulation and Ankle-Foot Orthosis on Gait Kinematics in Chronic Hemiparesis: A Randomized Controlled Trial.
Impaired ankle dorsiflexion in hemiparesis may be treated with ankle-foot orthosis or functional electrical stimulation. Semi-implanted selective functional electrical stimulation uses independent stimulations of deep and superficial peroneal nerves. The aim of this study was to compare gait kinematics using ankle-foot orthosis or semi-implanted selective functional electrical stimulation over 6 months in hemiparesis. Subjects with chronic hemiparesis, randomized into ankle-foot orthosis or semi-implanted selective functional electrical stimulation groups, underwent comfortable gait analysis without and with device OFF and ON, before, and 3 and 6 months after treatment onset. The effects of condition, visit and group on gait kinematics (analysis of variance; ANOVA) were analysed. A total of 27 subjects were included (ankle-foot orthosis, n = 13; semi-implanted selective functional electrical stimulation, n = 14). The only between-group difference in changes from OFF to ON conditions was a deteriorated ankle dorsiflexion speed with ankle-foot orthosis at month 6 (condition*group, p = 0.04; ankle-foot orthosis, -60%, p = 0.02; semi-implanted selective functional electrical stimulation, non significant). Both groups pooled, from OFF to ON gait speed (+ 0.07 m/s; + 10%), cadence (+ 4%), step length (+ 6%) and peak ankle dorsiflexion (+ 6°) increased, and peak ankle inversion (-5°) and peak knee flexion (-2°) decreased (p < 0.001); finally, peak knee flexion in the OFF condition increased (+ 2°, p = 0.03). Semi-implanted selective functional electrical stimulation and ankle-foot orthosis similarly impacted gait kinematics in chronic hemiparesis after 6 months of use. Ankle dorsiflexion speed in swing deteriorated markedly with ankle-foot orthosis.


Enhanced differentiation of HL-60 leukemia cells to macrophages induced by ciprofibrate.
Ciprofibrate, an hypolipidaemic peroxisome proliferator, induced differentiation of HL-60 cells. The effect was greatly potentiated by phorbol 12-myristate 13-acetate at a concentration where neither phorbol ester nor ciprofibrate alone had any effect on these cells. As occurs for HL-60 cell differentiation induced by high phorbol ester concentration, the ciprofibrate-induced phorbol ester-dependent differentiation of HL-60 cells proceeded through the monocytic/macrophage pathway and induced the phosphorylation of proteins with similar molecular weights suggesting that increased protein kinase C activity may be involved in the effect. The peroxisome proliferator-activated receptor (PPARalpha) transcription factor is expressed in HL-60 cells, but no changes were observed in its expression upon HL-60 cell differentiation.


Relationships between Lewy bodies and pale bodies in Parkinson's disease.
The prevalence of pale bodies and Lewy bodies was studied in the substantia nigra of 12 patients with typical Parkinson's disease (PD), in 5 patients with diffuse Lewy body disease (DLBD), and in a group of neurologically normal controls. Anti-ubiquitin antibodies labelled pale bodies and Lewy bodies in typical PD and DLBD, and there was a strong positive correlation between numbers of ubiquitin-immunoreactive pale bodies and Lewy bodies. BF10, a monoclonal antibody against a phosphate-dependent epitope of neurofilament 155-kDa polypeptide subunit, immunolabelled 57% of Lewy bodies and 15% of pale bodies in typical PD. Some pale bodies and Lewy bodies were seen in the substantia nigra of 2 of 5 neurologically normal, aged controls, probably representing "incidental PD". We conclude that there is a close relationship between pale bodies and typical Lewy bodies in the substantia nigra in clinical varieties of PD, and that these inclusions share antigenic determinants. If pale bodies and Lewy bodies reflect separate aspects of the cellular pathology in PD, their formation probably occurs in parallel. Alternatively, these observations may suggest that pale bodies represent a stage in the formation of Lewy bodies.


The PRB-dependent FOXO1/IGFBP-1 axis is essential for progestin to inhibit endometrial epithelial growth.
Progestin inhibits the growth of normal and cancerous endometria via the progesterone receptor (PR), but the distinct functions and signalings of PR subtypes have not been fully understood. The aim of the present study was to dissect the key pathways of progestin to inhibit endometrial epithelial growth. Immortalized endometrial epithelial cells (EM-E6/E7/TERT) with stable PRA or PRB expression were established and used for the experiments. In vitro growth inhibition by progestin was mainly observed in EM-E6/E7/TERT cells with PRB rather than those with PRA. RT-PCR assay confirmed that FOXO1, a key gene for progestin action, was up-regulated by progestin in a PRB-dependent manner. cDNA microarray analysis identified IGFBP-1, which contains FOXO1 binding sites on its promoter, to be induced by medroxyprogesterone acetate (MPA) in EM-E6/E7/TERT cells with PRB but not with PRA. siRNA knockdown of FOXO1 disturbed the induction of IGFBP-1 by MPA, while IGFBP-1 knockdown showed no effect on MPA-induced FOXO1 expression, indicating that FOXO1 is an upstream regulator of IGFBP-1. Luciferase reporter assays showed that MPA activated the IGFBP-1 promoter, which was cancelled by FOXO1 knockdown. Chromatin immunoprecipitation assay confirmed the in vivo binding of FOXO1 to the core promoter of IGFBP-1. IGFBP-1 knockdown significantly attenuated the growth inhibitory effects of MPA. The FOXO1/IGFBP-1 axis is essential for PRB-dependent growth inhibition of endometrial epithelial cells, offering a potential therapeutic clue to enhance the progestin effect.


Age-related differences in the response to myocardial ischemic stress.
Clinical experience indicates that the risk of reparative cardiac operations is increased in the neonatal period relative to that in older infants and children. Age-related differences in the susceptibility to myocardial ischemic dysfunction were evaluated by comparison of left ventricular function and metabolism in neonatal (mean age = 7 days) and weanling (mean age = 96 days) piglets. Six animals in each group supported on cardiopulmonary bypass were subjected to (1) 120 minutes of hypothermic crystalloid cardioplegic arrest (CP-120) and (2) 15 minutes of normothermic ischemic arrest (NA-15) after a 60 minute interval of reperfusion. Left ventricular systolic and diastolic function was measured after each intervention via endocardially implanted ultrasonic dimension crystals in a septolateral minor-axis position. In both groups, systolic dysfunction was evidenced by an increase in the dimension-axis intercept (p = 0.001), but not the slope of the end-systolic pressure-dimension relation. Left ventricular end-diastolic stiffness, expressed as left ventricular end-diastolic pressure versus Lagrangian strain, increased to a similar degree in both groups (p = 0.001). Adenosine triphosphate levels declined significantly (p = 0.001) in both groups in response to the ischemic interventions with no evident intergroup differences. Lactate levels increased significantly during the course of the experiment (p = 0.04); however, the increases were greater (p = 0.009) at all intervals in the neonatal group. This study demonstrates age-related metabolic differences in response to ischemia consistent with a greater dependence on glycolysis in neonatal myocardia. However, the fact that discriminating age-related differences in left ventricular function were not evident suggests that factors other than young age per se account for the increased surgical mortality in the neonatal period.


Biochemical analysis of Rabin8, the guanine nucleotide exchange factor for Rab8.
The Rab GTPases are master regulators of endosomal trafficking in eukaryotic cells. Among them, Rab8 plays an important role in tubulovesicular trafficking from the trans-Golgi network and recycling endosomes to the plasma membrane. Rab8 is activated by its guanine nucleotide exchange factor, Rabin8. In order to understand the molecular mechanisms that control endosomal recycling to the plasma membrane, it is pivotal to understand how Rabin8 is regulated in cells. Recently, biochemical and cell biological studies have identified several mechanisms for Rabin8 activation, which involves the relief of the intramolecular autoinhibition of Rabin8. Here we describe biochemical methods that we have used recently to study the activation of Rabin8.


DNMT1 mediates proliferation, migration and invasion of extravillous trophoblasts by regulating the methylation level of APLNR.
Proliferation, migration and invasion of extravillous trophoblasts (EVTs) play an important role in the progression of preeclampsia (PE). The purpose of this study was to investigate the molecular mechanism by which DNA methylase regulates the transcription level of APLNR and affects the phenotypic function of EVTs. PE mice model and H/R model in HTR8/Svneo cells were constructed. Clinical samples of normal pregnant women and PE patients were collected. Expression and methylation level of APLNR in vivo and in vitro were detected. ChIP-qPCR was used to detect the binding of DNA methyltransferase at the APLNR promoter. The expression of DNA methyltransferase 1 (DNMT1), NO and eNOS in vitro were detected. EVTs proliferation, migration and invasion in vitro were detected. In placental tissues or HTR8/Svneo cells of the PE model group, the expression of APLNR was reduced and APLNR methylation level was up-regulated. There was no significant difference in the APLNR expression in placental tissues between normal pregnant women and PE patients. H/R conditions only promote the binding of DNMT1 at the APLNR promoter. DNMT1 interference decreased the enrichment degree of DNMT1 in APLNR promoter region and up-regulated the mRNA and protein levels of APLNR in vivo and in vitro. The activation of APLNR by Elabela (ELA) can promote eNOS transcription, thereby promoting cell proliferation and NO level, while eNOS inhibitor can reverse this effect. DNMT1 down-regulation inhibted APLNR methylation level, promoted eNOS transcription, and promoted EVTs proliferation, migration and invasion, which could be revised by the interference of APLNR. DNMT1 promotes eNOS transcription by inhibting APLNR methylation level, and promotes EVTs proliferation, migration and invasion, thus providing a new and broad application prospect for PE treatment.


Paper-based ELISA to rapidly detect Escherichia coli.
Escherichia coli is a generic indicator of fecal contamination, and certain serotypes cause food- and water-borne illness such as O157:H7. In the clinic, detection of bacteriuria, which is often due to E. coli, is critical before certain surgical procedures or in cases of nosocomial infection to prevent further adverse events such as postoperative infection or sepsis. In low- and middle-income countries, where insufficient equipment and facilities preclude modern methods of detection, a simple, low-cost diagnostic device to detect E. coli in water and in the clinic will have significant impact. We have developed a simple paper-based colorimetric platform to detect E. coli contamination in 5h. On this platform, the mean color intensity for samples with 10(5)cells/mL is 0.118±0.002 (n=4), and 0.0145±0.003 (P<0.01⁎⁎) for uncontaminated samples. This technique is less time-consuming, easier to perform, and less expensive than conventional methods. Thus, paper-based ELISA is an innovative point-of-care diagnostic tool to rapidly detect E. coli, and possibly other pathogens when customized as appropriate, especially in areas that lack advanced clinical equipment.


Specific protein-protein interactions of calsequestrin with junctional sarcoplasmic reticulum of skeletal muscle.
Minor protein components of triads and of sarcoplasmic reticulum (SR) terminal cisternae (TC), i.e. 47 and 37 kDa peptides and 31-30 kDa and 26-25 kDa peptide doublets, were identified from their ability to bind 125I calsequestrin (CS) in the presence of EGTA. The CS-binding peptides are specifically associated with the junctional membrane of TC, since they could not be detected in junctional transverse tubules and in longitudinal SR fragments. The 31-30 kDa peptide doublet, exclusively, did not bind CS in the presence of Ca2+. Thus, different types of protein-protein interactions appear to be involved in selective binding of CS to junctional TC.


Increased mesolimbic cue-reactivity in carriers of the mu-opioid-receptor gene OPRM1 A118G polymorphism predicts drinking outcome: a functional imaging study in alcohol dependent subjects.
The endogenous opioid system is involved in the pathophysiology of alcohol-use disorders. Genetic variants of the opioid system alter neural and behavioral responses to alcohol. In particular, a single nucleotide polymorphism rs1799971 (A118G) in the mu-opioid receptor gene (OPRM1) is suggested to modulate alcohol-related phenotypes and neural response in the mesocorticolimbic dopaminergic system. Little is known about the clinical implications of these changes. The current study investigated the relationship of genotype effects on subjective and neural responses to alcohol cues and relapse in a sample of abstinent alcohol-dependent patients. Functional magnetic resonance imaging (fMRI) was used to investigate alcohol cue-reactivity and drinking outcome of 81 abstinent alcohol-dependent patients. G-allele carriers displayed increased fMRI cue-reactivity in the left dorsal striatum and bilateral insulae. Neural responses to alcohol cues in these brain regions correlated positively with subjective craving for alcohol and positive expectations of alcohol׳s effects. Moreover, alcohol cue-reactivity in the left dorsal striatum predicted time to first severe relapse. Current results show that alcohol-dependent G-allele carriers׳ increased cue-reactivity is associated with an increased relapse risk. This suggests that genotype effects on cue-reactivity might link the OPRM1 A118G risk allele with an increased relapse risk that was reported in earlier studies. From a clinical perspective, risk-allele carriers might benefit from treatments, such as neuro-feedback or extinction-based therapy that are suggested to reduce mesolimbic reactivity.


Selective modulation of NMDA responses by reduction and oxidation.
Electrophysiological responses to the glutamate analog N-methyl-D-aspartate (NMDA) measured in three different central neuronal preparations are subject to a novel modulatory mechanism: they are substantially potentiated after exposure to the disulfide reducing agent dithiothreitol, while oxidation with 5-5-dithiobis-2-nitrobenzoic acid decreases the magnitude of the response. Modification of the NMDA response by either oxidation or reduction does not appear to affect the pharmacological properties of the receptor-channel complex. Since we observe that the redox state of the native receptor-channel complex varies widely among neurons, an in vivo mechanism that can strongly regulate NMDA-activated functions by either reduction or oxidation may exist. In addition, these results suggest that it may be possible to design specific redox agents for characterizing the NMDA receptor-channel complex.


Can preterm deliveries be prevented?
Our hospital serves poor, inner-city women who have a 17% preterm delivery rate. Middle-class women in San Francisco at high risk for preterm delivery have benefited from an antepartum program which emphasized patient education and close follow-up. Using a controlled, randomized design, we are investigating the impact of similar interventions. Patients determined to be at high risk before 18 weeks' gestation on the basis of the Creasy system are randomly assigned to the Preterm Labor Prevention Clinic or serve as high-risk controls. Sixty-four women assigned to the Preterm Labor Prevention Clinic and 68 high-risk control women have been delivered of their infants. No significant differences were noted for the percentages of preterm infants, mean gestational age, or birth weight. Preterm rupture of the membranes accounted for 40% of preterm deliveries in all high-risk patients. Thirty percent of preterm births were indicated for maternal or fetal reasons. The remaining 30% represented failure of tocolytic therapy.


Patent Angiostrongylus mackerrasae infection in a black flying fox (Pteropus alecto).
A wild adult male black flying fox (Pteropus alecto) was presented unable to fly or hang strongly. Necropsy and histological examination revealed a severe pneumonia, with numerous Angiostrongylus mackerrasae in the pulmonary artery. The pulmonary parenchyma contained numerous eggs and rare larvae. This is the first report of patent Angiostrongylus infection in an accidental (i.e. non-Rattus) host species. It is also the first report of A. mackerrasae infection in an accidental host (including flying foxes). Worms recovered from cases of suspected angiostrongyliasis should be examined in morphological detail to ensure correct identification.


Prenatal Life Events, Maternal Employment, and Postpartum Depression across a Diverse Population in New York City.
This study examined racial/ethnic disparities in three core postpartum depression (PPD) symptoms, and identified specific predictors of PPD including sociodemographic variables, life stressors and maternal employment. White, African American, Hispanic, and Asian/Pacific Islander women from the New York City area (n = 3010) completed the 2009-2011 Pregnancy Risk Assessment Monitoring System. African American women were less likely to have PPD than White women. Maternal employment during the postpartum was associated with an increased likelihood of PPD for White women relative to women who were not employed. Life stressors and maternal employment should be considered as culturally contextualized factors related to postpartum depression.


"Assessment of the social influence and facilitating conditions that support nurses' adoption of hospital electronic information management systems (HEIMS) in Ghana using the unified theory of acceptance and use of technology (UTAUT) model".
Hospital electronic information management systems (HEIMS) are widely used in Ghana, and hence its performance must be carefully assessed. Nurses as clinical health personnel are the largest cluster of hospital staff and are the pillar of healthcare delivery. Therefore, they play a crucial role in the adoption and assessment of HEIMSs in Ghana. This report sought to assess the "Social Influence" (SI) and "Facilitating Conditions" (FC) that support Nurses' Acceptance of HEIMS in Ghana using the "Unified Theory of Acceptance and Use of Technology" (UTAUT) model. This study applied a non-experimental survey design. An electronic platform questionnaire on smartphones was used to collect data on 660 nurses. Statistically, AMOS Structural Equation Modelling (SEM) version 22.0 was employed to examine the research model. "Behavioral Intention" (BI) to HEIMS use was significantly predicted by SI and FC (p < 0.001). Notably, both SI and FC had an influence on nurses' use behavior (UB) with behavioral intention (BI) as the mediator, which explains a total of 42.1% variance in the intention of nurses to use HEIMS. Likewise, UB of HEIMS was also significantly predicted by SI (R2 = 43.2) and BI (R2 = 0.39.6) with both constructs explaining a total of 51.7% of the variance in nurses' acceptance to use HEIMS. Nurses' adoption of HEIMS in terms of the UB was influenced by SI and BI, whiles SI and FC had the strongest influence on BI (serving as mediator) of UB to adopt and use HEIMS among the nurses in Ghanaian hospitals.


Genetic Characterization of a Cohort of Italian Patients with Congenital Adrenal Hyperplasia Due to 21-Hydroxylase Deficiency.
Defects in the steroid 21-hydroxylase gene (CYP21A2) cause 21-hydroxylase deficiency (21OHD), the main cause of congenital adrenal hyperplasia (CAH). The disease shows a broad spectrum of clinical forms, ranging from severe or classical (salt wasting, SW, and simple virilizing, SV), to mild late onset or nonclassical (NC). 21OHD affects 1 in 15,000 in its severe classic form and 1 in 200-1000 in its mild NC form. There are many studies reporting the frequency of CYP21A2 pathogenic variants in different populations; however, few of them provide comprehensive information about Italian patients. Here, we present genetic results from a cohort of 245 unrelated Italian individuals with clinical diagnosis of CAH due to 21OHD. A specific polymerase chain reaction (PCR) protocol combined with Sanger sequencing was used for CYP21A2 analysis. The multiplex ligation-dependent probe amplification (MLPA) assay was employed for copy number variation (CNV) determination. One hundred fourteen (46.5%) of the index cases had the NC form, 57 (23.3%) had the SV form, and 74 (30.2%) presented the SW form of the disease. The most prevalent variant found in NC patients was the p.Val282Leu (51.3%), while the most frequent variants in the classical form were p.Ile173Asn (8.6%) and c.293-13C>G (26.0%). In our study, the frequency of large rearrangements was 15.3%, with CAH-X alleles representing 40% of all DEL/CONV. In addition, 12 alleles carried rare variants, and 1 had a novel variant p.(Arg342Gln). We observed phenotype-genotype correlation in 94.7% of cases. A complete concordance was observed in Groups 0 (enzyme activity completely impaired) where all patients had the SW form as expected. In Group A (0-1% residual enzyme activity), 78.4% of patients had the anticipated SW form while 21.6% were diagnosed with the SV form. Within Group B (~ 2% residual enzyme activity), 93.4% of patients exhibited SV form and 6.5% SW disease. Finally, 92.6% and 7.4% of patients belonging to Group C (enzyme partially impaired to ~ 20-60% residual activity) exhibited NC and SV phenotypes, respectively. This work, representing a comprehensive genetic study, expanded the CYP21A2 variants spectrum of Italian patients with 21OHD and could be helpful in prenatal diagnosis and genetic counseling.


The initial tangent of the aortic pressure increase is an estimate of left ventricular contractility in pigs.
The aim was, to identify an estimate of left ventricular contractility derived from the aortic pressure wave without load changing manoeuvres. For this purpose, left ventricular contractility was assessed with several aortic pressure wave form derived parameters and was compared to standard parameters of left ventricular contractility (conductance technique) in an experimental study. Measurements were taken during baseline, after beta-stimulation and after injection of a beta-antagonist. The initial and the secondary tangent, the area under the aortic pressure, and the stroke volume were correlated with the endsystolic elastance, a mainly load independent measure of left ventricular contractility: The initial tangent of the aortic pressure increase correlated significantly with the endsystolic elastance (r = 0.54, P < 0.05). The initial tangent of the aortic pressure increase was significantly increased from baseline at beta-stimulation (from 20.2 +/- 4.7 to 36.4 +/- 6.8 mmHg s(-1), P < 0.05) and decreased after injection of a beta-antagonist (from 20.2 +/- 4.7 to 12.3 +/- 2.0, P < 0.05). Thus, we conclude that the initial tangent of the aortic pressure increase is a valid estimate of left ventricular contractility in piglets.


US Public Health Preparedness for Zika and Other Threats Remains Vulnerable.
The unanticipated global outbreak of Zika virus infection is the most current but certainly not the last emerging infectious disease challenge to confront the US public heath system. Despite a number of such threats in recent years, significant gaps remain in core areas of public health system readiness. Stable, sustained investments are required to establish a solid foundation for achieving necessary national public health emergency preparedness and response capacity.


Crosstalk between small GTPases and polarity proteins in cell polarization.
Cell polarization is crucial for the development of multicellular organisms, and aberrant cell polarization contributes to various diseases, including cancer. How cell polarity is established and how it is maintained remain fascinating questions. Conserved proteins of the partitioning defective (PAR), Scribble and Crumbs complexes guide the establishment of cell polarity in various organisms. Moreover, GTPases that regulate actin cytoskeletal dynamics have been implicated in cell polarization. Recent findings provide insights into polarization mechanisms and show intriguing crosstalk between small GTPases and members of polarity complexes in regulating cell polarization in different cellular contexts and cell types.


[Sanitary and epidemiological examination of foods under carrying sanitary and epidemiological surveillance on the customer border and at in the territory of the Customers Union].
The article concerns peculiarities of sanitary and epidemiological and hygienic examination of foods under carrying of sanitary-epidemiologic surveillance on the customer border and of the custom territory of the Customers Union of the Russian Federation, Kazakhstan and Byelorussia.


Psychiatric manifestations of autoimmune disorders.
Psychiatric symptoms are common to many autoimmune disorders. Patients often will have mood disorders, anxiety, cognitive deficits, delirium, and psychosis. These symptoms may reflect the direct or indirect effect of the autoimmune disorder on the central nervous system, may be related to medications used to treat the disorder, or may be a direct psychologic impact from suffering with the autoimmune disorder. Accurately recognizing the psychiatric component and generating a differential diagnosis is a complex task for the treating physician. Treatment of the psychiatric component to the disorder often will include addressing steroid induced side effects, psychotropic medications, psychotherapy, patient and family education, and a strong physician-patient relationship.


Human hepatic mitochondria generate reactive oxygen species and undergo the permeability transition in response to hydrophobic bile acids.
Hydrophobic bile acids accumulate in the liver during cholestasis and are believed to cause hepatocellular necrosis and apoptosis in part through induction of the mitochondrial permeability transition (MPT) and the mitochondrial generation of oxidative stress. The purpose of this study was to determine if human hepatic mitochondria respond to bile acids in this manner. The MPT was measured spectrophotometrically and morphologically in normal human liver mitochondria exposed to glycochenodeoxycholic acid (GCDC) with and without cyclosporin A, an inhibitor of the MPT, antioxidants, and tauroursodeoxycholic acid (TUDC). Hydroperoxide generation was measured by dichlorofluorescein fluorescence. Cytochrome c and apoptosis-inducing factor were assessed by immunoblotting. GCDC induced the MPT in a dose-dependent manner, which was inhibited by cyclosporin A, alpha-tocopherol, beta-carotene, idebenone, and TUDC. GCDC stimulated reactive oxygen species generation and release of cytochrome c and apoptosis-inducing factor, which were significantly inhibited by the antioxidants, cyclosporin A, and TUDC. Mitochondrial pathways of cell death are stimulated in human hepatic mitochondria exposed to GCDC consistent with the role of mitochondrial dysfunction in the pathogenesis of cholestatic liver injury. These results parallel those reported in rodents, supporting the extrapolation of mechanistic studies of bile acid toxicity from rodent to humans.


The Salmonella PmrAB regulon: lipopolysaccharide modifications, antimicrobial peptide resistance and more.
Microbes are able to sense and respond to their environment primarily through the use of two-component regulatory systems. Many of these systems activate virulence-factor expression and are regulated by host-derived signals, having evolved to control gene expression at the key time and place for optimal establishment and maintenance of infection. Salmonella spp. are enteric pathogens that are able to survive both within host macrophages during systemic spread and killing by innate immune factors at intestinal mucosal surfaces. This review focuses on a key mechanism of pathogenesis that involves the PmrA-PmrB two-component system, which is activated in vivo by direct or indirect means and regulates genes that modify lipopolysaccharide, aiding survival in host (and non-host) environments.


Engineered dynamic distribution of malonyl-CoA flux for improving polyketide biosynthesis in Komagataella phaffii.
Malonyl-CoA is a basic but limited precursor for the biosynthesis of various bioactive compounds and life-supporting fatty acids in cells. This study develops a biosynthetic system to dynamically redirect malonyl-CoA flux and improve production of malonyl-CoA derived polyketide (6-MSA) in Komagataella phaffii. A synthetic regulatory protein fusing a yeast activator Prm1 with a bacterial repressor FapR was proved to work with a hybrid promoter (-7)fapO-cPAOX1 and activate gene expression. Expression mode by the Prm1-FapR/(-7)fapO-cPAOX1 device was not affected by intracellular malonyl-CoA levels. Further, 9 promoter variants of PGAP with insertion of fapO at various sites were tested with the Prm1-FapR. It generated a biosensor of Prm1-FapR/PGAP-(+2)fapO with regulation behavior of malonyl-CoA-low-level repression/high-level derepression. Both devices were subsequently integrated into a single cell, for which fatty acid synthesis module was driven by Prm1-FapR/PGAP-(+2)fapO but 6-MSA synthesis module was expressed by Prm1-FapR/(-7)fapO-cPAOX1. The integrated system allowed continuous polyketide synthesis but malonyl-CoA-high-level "on"/low-level "off" fatty acid synthesis. This design finally increased 6-MSA production capacity by 260 %, proving the positive effects of dynamic malonyl-CoA distribution to the target compounds. It provides a new strategy for synthesis of malonyl-CoA derived compounds in eukaryotic chassis hosts.


Development of a clinically relevant ovarian cancer model incorporating surgical cytoreduction to evaluate treatment of micro-metastatic disease.
Mouse models of ovarian cancer commonly transfer large numbers of tumor cells into the peritoneal cavity to establish experimental metastatic disease, which may not adequately model early metastatic spread from a primary tumor site. We hypothesized we could develop an ovarian cancer model that predictably represents micro-metastatic disease. Murine ID8VEGF ovarian cancer cells were transduced to express enhanced luciferase (eLuc) to enable intravital detection of microscopic disease burden and injected beneath the ovarian bursa of C57Bl/6 mice. At 6 or 10 weeks after orthotopic injection, when mice had detectable metastases, hysterectomy and bilateral salpingo-oophorectomy was performed to remove all macroscopic disease, and survival monitored. Immunohistochemistry and gene expression profiling were performed on primary and metastatic tumors. eLuc-transduced ID8VEGF cells were brighter than cells transduced with standard luciferase, enabling in vivo visualization of microscopic intra-abdominal metastases developing after orthotopic injection. Primary surgical cytoreduction removed the primary tumor mass but left minimal residual disease in all mice. Metastatic sites that developed following orthotopic injection were similar to metastatic human ovarian cancer sites. Gene expression and immune infiltration were similar between primary and metastatic mouse tumors. Surgical cytoreduction prolonged survival compared to no surgery, with earlier cytoreduction more beneficial than delayed, despite micro-metastatic disease in both settings. Mice with primary ovarian tumors established through orthotopic injection develop progressively fatal metastatic ovarian cancer, and benefit from surgical cytoreduction to remove bulky disease. This model enables the analysis of therapeutic regimens designed to target and potentially eradicate established minimal residual disease.


Fine-scale mapping of the 4q24 locus identifies two independent loci associated with breast cancer risk.
A recent association study identified a common variant (rs9790517) at 4q24 to be associated with breast cancer risk. Independent association signals and potential functional variants in this locus have not been explored. We conducted a fine-mapping analysis in 55,540 breast cancer cases and 51,168 controls from the Breast Cancer Association Consortium. Conditional analyses identified two independent association signals among women of European ancestry, represented by rs9790517 [conditional P = 2.51 × 10(-4); OR, 1.04; 95% confidence interval (CI), 1.02-1.07] and rs77928427 (P = 1.86 × 10(-4); OR, 1.04; 95% CI, 1.02-1.07). Functional annotation using data from the Encyclopedia of DNA Elements (ENCODE) project revealed two putative functional variants, rs62331150 and rs73838678 in linkage disequilibrium (LD) with rs9790517 (r(2) ≥ 0.90) residing in the active promoter or enhancer, respectively, of the nearest gene, TET2. Both variants are located in DNase I hypersensitivity and transcription factor-binding sites. Using data from both The Cancer Genome Atlas (TCGA) and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC), we showed that rs62331150 was associated with level of expression of TET2 in breast normal and tumor tissue. Our study identified two independent association signals at 4q24 in relation to breast cancer risk and suggested that observed association in this locus may be mediated through the regulation of TET2. Fine-mapping study with large sample size warranted for identification of independent loci for breast cancer risk.


2- and 2,7-Substituted para-N-Methylpyridinium Pyrenes: Syntheses, Molecular and Electronic Structures, Photophysical, Electrochemical, and Spectroelectrochemical Properties and Binding to Double-Stranded (ds) DNA.
Two N-methylpyridinium compounds and analogous N-protonated salts of 2- and 2,7-substituted 4-pyridyl-pyrene compounds were synthesised and their crystal structures, photophysical properties both in solution and in the solid state, electrochemical and spectroelectrochemical properties were studied. Upon methylation or protonation, the emission maxima are significantly bathochromically shifted compared to the neutral compounds, although the absorption maxima remain almost unchanged. As a result, the cationic compounds show very large apparent Stokes shifts of up to 7200 cm-1 . The N-methylpyridinium compounds have a single reduction at ca. -1.5 V vs. Fc/Fc+ in MeCN. While the reduction process was reversible for the 2,7-disubstituted compound, it was irreversible for the mono-substituted one. Experimental findings are complemented by DFT and TD-DFT calculations. Furthermore, the N-methylpyridinium compounds show strong interactions with calf thymus (ct)-DNA, presumably by intercalation, which paves the way for further applications of these multi-functional compounds as potential DNA-bioactive agents.


The role of bone scan and radiography in the diagnostic evaluation of suspected pedal osteomyelitis.
The authors reviewed the three-phase bone scans and radiographs of 24 patients with suspected pedal osteomyelitis who also had histologic confirmation of the diagnosis. Twenty patients had a pedal ulcer, cellulitis, or necrosis. Sensitivity and specificity of bone scanning were 70% and 43% respectively. Sensitivity and specificity of radiography were 70% and 50% respectively. The non-invasive diagnosis of pedal osteomyelitis remains problematic due to the poor specificity of bone scans and radiographs.


Fluorescent sensor for Cr(VI) based in functionalized silicon quantum dots with dendrimers.
Highly luminescent nanoparticles based in Silicon quantum dots, coated by hydroxyl PAMAM dendrimer (PAMAM-OH) of 5th generation, were obtained by one step process by hydrothermal treatment of 3-Aminopropyl)triethoxysilane (APTES) in aqueous solution. Previous to the optimization of the synthesis procedure, different dendritic molecules of 5th generation were tested to obtain the most intense fluorescence signal. The influence of different parameters such ratio APTES/PAMAM-OH, pH and ionic strength on the fluorescence intensity was studied. The fluorescence spectra showed maximum excitation and emission wavelengths at 370 and 446 nm, respectively. The obtained silicon nanoparticles (SiQDs@PAMAM-OH) were characterized by TEM, DLS and XPS, and were found to detect selectively Cr(VI) in aqueous solutions at 2.7 μM level of detection, sensitivity of 0.2 μM with a RSD of 0.16% (n=10). To study the feasibility of the proposed system for Cr(VI) detection, it was tested in real electrochemical solution bath and a tanning effluent obtained from electrochemical industry and with two certified waters, demonstrating promising outcomes as nano-sensor.


Design and synthesis of N-(5-chloro-2,4-dihydroxybenzoyl)-(R)-1,2,3,4-tetrahydroisoquinoline-3-carboxamides as novel Hsp90 inhibitors.
Heat shock protein 90 (Hsp90) is an attractive chemotherapeutic target for antitumor drug development. Herein, we reported the design and synthesis of two series of novel N-(5-chloro-2,4-dihydroxybenzoyl)-1,2,3,4-tetrahydroisoquinoline-3- carboxamides as Hsp90 inhibitors using (S)-Tic (1,2,3,4-tetrahydroisoquinoline-3- carboxylic acid) (A1-13) and (R)-Tic (B1-13) as scaffold, respectively. Cellular thermal shift assay (CETSA) screening showed that compounds B1-13 with (R)-Tic scaffold exhibited potent ability to stabilize Hsp90α. Compound B7 showed not only the most potent ability to induce thermal stabilization of Hsp90α but also the strongest cytotoxicity. The IC50 values of B7 were 0.98 μM and 1.74 μM against the proliferation of human breast cancer MDA-MB-231 and human cervical cancer HeLa cell lines, respectively. Moreover, CETSA melt and ITDRFCETSA (isothermal dose-response fingerprint) curves confirmed that B7 bound to Hsp90α in 293T cells. Western blotting results indicated that B7 induced the degradation of Hsp90 clients CDK4, Her2, Cdc-2 and C-raf. In addition, docking and Molecular dynamics (MD) refinement of the B7-Hsp90 complex showed that the binding model of B7 to Hsp90 was similar with that of 8-benzyladenines. The overall properties warrant compound B7 a promising lead for the development of Hsp90 inhibitor antitumor drugs.


Topographical distribution and characterization of epithelial cells and intraepithelial lymphocytes in the human ocular mucosa.
The conjunctiva plays a key role in the protection of the ocular surface by initiating and regulating immune responses. In this study, we analyze the relative proportion of intraepithelial lymphocytes (IELs), apoptotic cells, and proliferative state in three different topographical regions of the normal human conjunctiva. Superior tarsal, superior bulbar, and inferior tarsal-bulbar-fornical conjunctival cells were collected by brush cytology from 63 healthy donors. Flow cytometry analysis showed higher levels of CD3⁺ and CD8⁺ IELs in both upper tarsal and bulbar conjunctiva than in the inferior tarsal-bulbar-fornix, where the CD19⁺ B cells were increased. For all zones two different cell populations (by cell size and complexity) were present in the apoptosis assay. The more complex cells were reduced within the inferior tarsal-bulbar-fornix when compared with the superior bulbar and tarsal areas. Less complex cells were more predominant in the inferior conjunctiva and were mainly alive. The mean proliferation index of the conjunctival epithelium was significantly lower in the superior bulbar conjunctiva than in superior tarsal and inferior fornical conjunctivas. These findings suggest that each topographical zone from normal human conjunctiva has a unique profile of immunophenotype, viability, and proliferative state that could be related to a differentiated regional functionality.


COVID-19 case of ventilator-induced lung injury on extracorporeal membrane oxygenation: Physicians' clinical struggle and ethical conflict in a novel pandemic.
Resource scarcity was concerned in the initial surge of the COVID-19 pandemic. To open slots for Extracorporeal Membrane Oxygenation (ECMO), we tried ECMO weaning allowing invasive ventilation in a 66-year-old male with severe COVID-19, backfiring as ventilator-induced lung injury. We will discuss ethical conflict in pandemics in this report.


Estimation of Partially Specified Dynamic Spatial Panel Data Models with Fixed-Effects.
This paper studies estimation of a partially specified spatial panel data linear regression with fixed-effects. Under the assumption of strictly exogenous regressors and strictly exogenous spatial weighting matrix, the unknown parameter is estimated by applying the instrumental variable estimation. Under some sufficient conditions, the proposed estimator for the finite dimensional parameter is shown to be root-N consistent and asymptotically normally distributed; The proposed estimator for the unknown function is shown to be consistent and asymptotically distributed as well, though at a rate slower than root-N. Consistent estimators for the asymptotic variance-covariance matrices of both estimators are provided. The results can be generalized to several spatial weighting matrice and spatial matrix which vary with time. The simulation results suggest that the proposed approach has some practical value.


Eosinophilic esophagitis strongly linked to chronic rhinosinusitis.
To determine the relative risk of having eosinophilic esophagitis (EoE) coexist with chronic rhinosinusitis (CRS) in probands and their families using the Utah Population Database (UPDB). This retrospective observational cohort study with population-based matched controls utilized the UPDB, a genealogical database linked to medical records. It was used to identify CRS and EoE patients diagnosed at any age between 2008 and 2012. The familial risks of an EoE diagnosis (International Classification of Diseases, Ninth Revision, Clinical Modification code 530.13) in CRS probands, and vice versa, and their first- through fifth-degree relatives and spouses were calculated using logistic regression models in comparison to controls randomly selected from the Utah population and individually matched 6:1 on sex and birth year. Probands with CRS demonstrated a 3.4-fold increased risk of having EoE themselves (P < 10(-15) ). First-degree relatives (parents, siblings, and children) of CRS probands had a 1.5-fold increased risk of having EoE (P < 10(-4) ), whereas more distant relatives did not show a significant increased risk. Spouses of probands had a 1.4-fold increased risk of having EoE (P = 0.055). Conversely, risk estimates of having CRS in EoE probands were consistent. We observed an increased risk of comorbid EoE in patients with CRS and their families. An association between CRS and EoE as comorbid conditions suggests that a familial component is contributing to the etiology of both diseases. Further analyses regarding the pathophysiology of the development of CRS in these specific patients will lead to a better understanding of both disease processes and may help target therapy. 3b. Laryngoscope, 126:1279-1283, 2016.


Smoking and age-related macular degeneration: a review of association.
Age-related macular degeneration (AMD) is the leading cause of severe and irreversible vision loss in the Western world. As there is no effective treatment for all types of AMD, identifying modifiable risk factors is of great importance. This review evaluates the epidemiological evidence associating smoking with AMD. Systematic review of published epidemiological studies evaluated against established criteria for evidence of a causal relationship. In total, 17 studies (cross-sectional studies, prospective cohort studies, and case-control studies) were included in the review. A total of 13 studies found a statistically significant association between smoking and AMD with increased risk of AMD of two- to three-fold in current-smokers compared with never-smokers. Five studies found no association between smoking and AMD. There was also evidence of dose-response, a temporal relationship and reversibility of effect. The literature review confirmed a strong association between current smoking and AMD, which fulfilled established causality criteria. Cigarette smoking is likely to have toxic effects on the retina. In spite of the strength of this evidence, there appears to be a lack of awareness about the risks of developing eye disease from smoking among both healthcare professionals and the general public.


Naturally available diatomite and their surface modification for the removal of hazardous dye and metal ions: A review.
The presence of toxic pollutants such as dyes and metal ions at higher concentrations in water is very harmful to the environment. Removal of these pollutants using diatomaceous earth or diatomite (DE) and surface-modified DE has been extensively explored due to their excellent physio-chemical properties and low cost. Therefore, naturally available DE being inexpensive, their surface modified adsorbents could be one of the potential candidates for the wastewater treatment in the future. In this context, the current review has been summarized for the removal of both pollutants i.e., dyes and metal ions by surface-modified DE using the facile adsorption process. In addition, this review is prominently focused on the various modification process of DE, their cost-effectiveness; the physio-chemical characteristics and their maximum adsorption capacity. Further, real-time scenarios of reported adsorbents were tabulated based on the cost of the process along with the adsorption capacity of these adsorbents.


Identification of structural brain alterations in adolescents with depressive symptomatology.
Depressive symptoms can emerge as early as childhood and may lead to adverse situations in adulthood. Studies have examined structural brain alternations in individuals with depressive symptoms, but findings remain inconclusive. Furthermore, previous studies have focused on adults or used a categorical approach to assess depression. The current study looks to identify grey matter volumes (GMV) that predict depressive symptomatology across a clinically concerning sample of adolescents. Structural MRI data were collected from 338 clinically concerning adolescents (mean age = 15.30 SD=2.07; mean IQ = 101.01 SD=12.43; 132 F). Depression symptoms were indexed via the Mood and Feelings Questionnaire (MFQ). Freesurfer was used to parcellate the brain into 68 cortical regions and 14 subcortical regions. GMV was extracted from all 82 brain areas. Multiple linear regression was used to look at the relationship between MFQ scores and region-specific GMV parameter. Follow up regressions were conducted to look at potential effects of psychiatric diagnoses and medication intake. Our regression analysis produced a significant model (R2 = 0.446, F(86, 251) = 2.348, p < 0.001). Specifically, there was a negative association between GMV of the left parahippocampal (B = -0.203, p = 0.005), right rostral anterior cingulate (B = -0.162, p = 0.049), and right frontal pole (B = -0.147, p = 0.039) and a positive association between GMV of the left bank of the superior temporal sulcus (B = 0.173, p = 0.029). Follow up analyses produced results proximal to the main analysis. Altered regional brain volumes may serve as biomarkers for the development of depressive symptoms during adolescence. These findings suggest a homogeneity of altered cortical structures in adolescents with depressive symptoms.