Co-Treatment With Verapamil and Curcumin Attenuates the Behavioral Alterations Observed in Williams-Beuren Syndrome Mice by Regulation of MAPK Pathway and Microglia Overexpression.
Williams-Beuren syndrome (WBS) is a rare neurodevelopmental disorder characterized by a distinctive cognitive phenotype for which there are currently no effective treatments. We investigated the progression of behavioral deficits present in WBS complete deletion (CD) mice, after chronic treatment with curcumin, verapamil, and a combination of both. These compounds have been proven to have beneficial effects over different cognitive aspects of various murine models and, thus, may have neuroprotective effects in WBS. Treatment was administered orally dissolved in drinking water. A set of behavioral tests demonstrated the efficiency of combinatorial treatment. Some histological and molecular analyses were performed to analyze the effects of treatment and its underlying mechanism. CD mice showed an increased density of activated microglia in the motor cortex and CA1 hippocampal region, which was prevented by co-treatment. Behavioral improvement correlated with the molecular recovery of several affected pathways regarding MAPK signaling, in tight relation to the control of synaptic transmission, and inflammation. Therefore, the results show that co-treatment prevented behavioral deficits by recovering altered gene expression in the cortex of CD mice and reducing activated microglia. These findings unravel the mechanisms underlying the beneficial effects of this novel treatment on behavioral deficits observed in CD mice and suggest that the combination of curcumin and verapamil could be a potential candidate to treat the cognitive impairments in WBS patients.


How Do Basic Human Values Affect Self-Care of Type 2 Diabetes Patients? A Multicentre Observational Study.
In type 2 diabetes mellitus (T2DM) patients, human values have been identified as having a possible impact on self-care. Despite this, basic human values have never been described in the T2DM population and the association between basic human values and self-care has not been assessed. The aims of this study were to describe basic human values and self-care, and investigate the associations between basic human values and self-care in adults with T2DM. The study was cross-sectional multicentre with T2DM patients (n = 390). Sociodemographic and clinical data were collected from charts, and two questionnaires were administered. The Portrait Values Questionnaire (PVQ) measured basic human values and allowed us to group patients into four groups based on the value profile: (A) Openness to Change and Self-Transcendence, (B) Conservation and Self-Transcendence, (C) Conservation and Self-Enhancement, and (D) Openness to Change and Self-Enhancement. The Self-care of Diabetes Inventory measured self-care maintenance, self-care monitoring, self-care management, and self-care confidence, according to the Middle-Range Theory of Self-Care of Chronic Illness. Multiple regression models were used to assess associations between the basic human values and self-care. Group D (Openness to Change and Self-Enhancement) performed significantly lower self-care maintenance (p = .024) and self-care monitoring (p = .031) when compared with Group A, and lower self-care monitoring (p = .008) and self-care management (p = .018) when compared with Group B. Group D showed significantly lower self-care monitoring (p = .027) when compared with Group C. People with T2DM who value more self-enhancement and openness to change might be more prone to perform worse self-care compared with those who value conservation and self-transcendence. Clinicians should consider this when designing interventions aimed at improving self-care behaviors in patients with T2DM.


Simultaneous determination and pharmacokinetics study of three triterpenoid saponins in rat plasma by ultra-high-performance liquid chromatography tandem mass-spectrometry after oral administration of Astragalus Membranaceus leaf extract.
A selective and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of three triterpenoid saponins isolated from Astragalus membranaceus leaf extract. In this article, a method for simultaneous determination of Huangqiyenin A, Huangqiyenin E, and Huangqiyenin K was established for the first time. The method was successfully applied to the pharmacokinetic study of Astragalus membranaceus leaf extract after oral administration. Liquid-liquid extraction was applied to plasma sample preparation. Multiple reaction monitoring mode with an electrospray ion source in positive electrospray ionization was chosen to quantify the analytes. Chromatographic separation was performed on a Waters HSS T3 column, using gradient elution with a mobile phase composed of acetonitrile and 5 mM ammonium acetate/water. The pharmacokinetic results showed that all three compounds had the characteristics of rapid absorption-slow metabolism trend. The time of maximum plasma concentration of Huangqiyenin A is higher than Huangqiyenin E and Huangqiyenin K. And the maximum plasma concentration of Huangqiyenin A is higher as well. The pharmacokinetic results revealed the pharmacokinetic characteristics of the three analytes in rat plasma, which could provide a helpful reference for the further study of Astragalus membranaceus leaf extract.


Progressive familial intrahepatic cholestasis among the Arab population in Israel.
Progressive familial intrahepatic cholestasis, which constitutes a heterogeneous group of imperfectly delineated syndromes and appears to be inherited as an autosomal recessive condition, has not been hitherto reported from the Middle East, in spite of the high rate of consanguineous marriage in this region. Sixteen affected children from six Israeli Arab families were evaluated over 30 years. All were born to consanguineous parents. Jaundice appeared during the first 3 weeks of life in 15 babies. When first referred, 10 had hepatomegaly and nine had splenomegaly. A progression toward cirrhosis was the rule. Serum levels of conjugated bilirubin, liver enzymes, and alkaline phosphatase were raised; gamma-glutamyl transpeptidase levels were normal in all three infants in whom it was examined, but elevated in two siblings of another family at ages 2 and 3 years. No abnormal bile acids were detected in the serum and urine of patients. Histologic examination of the liver showed giant-cell transformation, paucity of intrahepatic bile ducts, cholestasis, fibrosis, or cirrhosis. The pattern of liver pathology differed at times among affected members within the same family. Therapeutic trials with phenobarbital, cholestyramine, or ursodeoxycholic acid were ineffective. Survival of the patients was from 5 to 18 months in four families; in the other two families, three children received liver transplants, and one is awaiting liver transplantation. Progressive familial intrahepatic cholestasis should be included in the differential diagnosis of infants with cholestatic jaundice of unknown etiology, especially those born to consanguineous Arab parents.


Cinemeducation: an innovative approach to teaching psychosocial medical care.
This article discusses the use of clips of popular movies on videotape to educate family practice residents in the psychosocial aspects of medical care. Video clips anchor residents' insights about patients from clinical practice and illustrate family life cycle issues and Diagnostic and Statistical Manual of Mental Disorders-Revised diagnoses. Movies capture learners' attention, expose residents to diverse life-styles, engage the humanistic side of physicians, and imprint powerful pictorial images in memory. Teaching with film clips is time efficient and provides emotionally engaging experiences for faculty and residents. Selected films are reviewed for suitable clips and then incorporated into 1-hour teaching conferences. Successful "cinemeducation" requires appropriate films on videocassette, a VCR with a real-time counter, a television screen in clear view of all class members, audio speakers with sufficient volume to hear dialogue without distortion, and a willingness to be open to the emotional impact of movies. A detailed list identifies movie scenes that can be readily incorporated into the psychosocial teaching program of any medical education curriculum.


Urgent arterial embolization of ruptured renal angiomyolipoma.
The most undesired complication of renal angiomyolipoma (AML) is bleeding. Because of tumor rupture, the bleeding can spread to the retroperitoneal field and can be severe enough to be life threatening. We report a case of retroperitoneal hemorrhage caused by a ruptured AML that was successfully treated with transarterial embolization with N-butyl cyanoacrylate.


Several anthropometric measurements and breast cancer risk: results of the E3N cohort study.
To investigate the association between various anthropometric characteristics and breast cancer. Longitudinal prospective cohort study. Follow-up between 1995 and 2000. In total, 69 116 women (age: 45-70 years; mean follow-up: 3.6 years), 275 premenopausal and 860 postmenopausal incident invasive breast cancers. Self-reported height, weight, breast, thorax, waist and hip circumferences and calculated body mass index (BMI) and waist-to-hip ratio (WHR) at baseline. A slight increase in risk with increasing height was found. Weight, BMI, thorax and waist circumferences and WHR were negatively related to breast cancer risk among premenopausal women. The relationships became non significant after additional adjustment for BMI. An increased risk of premenopausal breast cancer with an android body shape (WHR>0.87) might possibly be confined to obese women. Among postmenopausal women, all anthropometric measurements of corpulence were positively associated with breast cancer risk but became non significant after additional adjustment for BMI. No difference in risk of postmenopausal breast cancer according to HRT use was observed. The study confirmed that adiposity was negatively associated to premenopausal breast cancer risk and positively associated to postmenopausal breast cancer risk. Further studies will be needed to specify clearly the association between WHR and breast cancer risk, particularly before menopause.


Phosphine-Free Synthesis of Metal Chalcogenide Quantum Dots by Directly Dissolving Chalcogen Dioxides in Alkylthiol as the Precursor.
Semiconductor quantum dots (QDs) are competitive emitting materials in developing new-generation light-emitting diodes (LEDs) with high color rendering and broad color gamut. However, the use of highly toxic alkylphosphines cannot be fully avoided in the synthesis of metal selenide and telluride QDs because they are requisite reducing agents and solvents for preparing chalcogen precursors. In this work, we demonstrate the phosphine-free preparation of selenium (Se) and tellurium (Te) precursors by directly dissolving chalcogen dioxides in the alkylthiol under the mild condition. The chalcogen dioxides are reduced to elemental chalcogen clusters, while the alkylthiol is oxidized to disulfides. The chalcogen clusters further combine with the disulfides, generating dispersible chalcogen precursors. The resulting chalcogen precursors are suitable for synthesizing various metal chalcogenide QDs, including CdSe, CdTe, Cu2Te, Ag2Te, PbTe, HgTe, and so forth. In addition, the precursors are of high reactivity, which permits a shorter QD synthesis process at lower temperature. Owing to the high quantum yield (QYs) and easy tunability of the photoluminescence (PL), the as-synthesized QDs are further employed as down-conversion materials to fabricate monochrome and white LEDs.


Probing the Binding Modes of a Multidomain Protein to Lipid-based Nanoparticles by Relaxation-based NMR.
The interactions of two model multidomain proteins-covalently linked diubiquitins, Ub2-with lipid-based nanoparticles have been quantitatively probed by the measurements of NMR lifetime line broadening, ΔR2. By combined analysis of ΔR2 profiles arising from interactions with liposomes of varying sizes, an approach recently developed for the characterization of interactions of monoubiquitin with liposomes, we determine how the parameters of exchange (liposome binding) and dynamics of each individual domain of Ub2 on the surface of liposomes change when the domains are covalently attached to one another by a flexible linker. Two different covalent linkages were used: K63-linked and K48-linked Ub2. The possibility of three distinct modes of binding of Ub2 to liposomes requires the introduction of simple but important modifications to the strategy of analysis originally developed for monoubiquitin.


Controlling quantum-dot light absorption and emission by a surface-plasmon field.
The possibility for controlling both the probe-field optical gain and absorption, as well as photon conversion by a surface-plasmon-polariton near field is explored for a quantum dot located above a metal surface. In contrast to the linear response in the weak-coupling regime, the calculated spectra show an induced optical gain and a triply-split spontaneous emission peak resulting from the interference between the surface-plasmon field and the probe or self-emitted light field in such a strongly-coupled nonlinear system. Our result on the control of the mediated photon-photon interaction, very similar to the 'gate' control in an optical transistor, may be experimentally observable and applied to ultra-fast intrachip/interchip optical interconnects, improvement in the performance of fiber-optic communication networks, and developments of optical digital computers and quantum communications.


alpha-1-Antitrypsin phenotypes in hepatocellular carcinoma.
alpha-1-Antitrypsin deficiency due to homozygous Pi ZZ state is reported to be associated with cirrhosis and hepatocellular carcinoma (HCC); however, the role of heterozygous Pi Z state is not definitely known. In order to investigate the possible association, we studied the phenotypic distribution of alpha-1-antitrypsin variants (Pi) in 124 cases of HCC. Two thousand ten normal American Red Cross blood donors were studied as controls. Twelve patients with HCC had aberrant phenotypes, an incidence of 9.67% as compared to 8.36% among normal controls. Nine of 12 patients with HCC with aberrant Pi type had cirrhosis; 5 of the 9 had cirrhosis due to hepatitis B virus; 2 of the 9 had alcoholic liver disease with cirrhosis, and 2 had cryptogenic cirrhosis. The three patients with HCC arising in noncirrhotic livers who also had aberrant Pi phenotypes, had a relatively rare variety of HCC called fibrolamellar type. Z gene was found in five patients: all five were MZ. Incidence of MZ phenotype in HCC was similar to that of the normal control population (4.0% in HCC and 2.9% in the controls). However, 3 of 5 MZ were associated with fibrolamellar HCC. Another aberrant phenotype found among the patients with HCC was MF (fast moving) which occurred with an incidence of 2.41% as compared to none in the control group. In conclusion, we found no significant increase in the incidence of Z gene among 124 patients with HCC as compared to the normal population.


Incidence and nature of horse-riding injuries. A one-year prospective study.
A prospective one-year study of horse-riding accidents was performed in an area with a very high frequency of amateur riding. Totally 174 patients with riding injuries were registered--66% children--making an incidence of only 0.7 per thousand riding occasions. However, many of these injuries were of a severe nature, one leading to death and 27 of the patients reporting persisting symptoms at follow-up one year after the accident. Eleven per cent of the accidents lead to cerebral injuries and 44% to fractures, mainly of the upper extremities. Eleven per cent required hospitalization and 17% operation under general anaesthesia. Clarification of the circumstances concerning the accidents indicated that many of the severe injuries could probably have been avoided by use of better safety equipment.


Structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart. I: the conus valves and the subendocardium.
Sturgeons are bony fish that retain structural traits typical of the more primitive Chondrostei. From an evolutionary viewpoint, sturgeons are considered relic fish. However, they show remarkable ecological plasticity and are well adapted to contemporary environmental conditions. Although development of the cardiovascular system is critical for all organs and systems, and is affected by evolutionary changes, the structure of the sturgeon heart has been mostly overlooked. This is also true for the conus arteriosus, which, as in Chondrostei, is endowed with several rows of valves and a layer of contractile myocardium. This work reports on the structure of the valves, the endocardium, and the subendocardium of the conus arteriosus of the sturgeon (Acipenser naccarii) heart. It is part of a broader study that aims to cover the entire structure of the sturgeon heart. The conus arteriosus of 15 A. naccarii hearts, ranging in age from juveniles to sexually-differentiated adults, has been studied by conventional light, transmission (TEM), and scanning electron microscopy (SEM). In addition, maceration of the soft tissues with NaOH, and actin localization by fluorescent phalloidin has been used. The conus is a tubular chamber that arises from the right ventricular side and presents two constrictions at the conus-ventricle and conus-aorta junctions. The conus is endowed with three rows of valves: one distal and two proximal. The segment of the conus located between the distal and the two proximal rows is devoid of valvular structures. The distal row has four leaflets, while the two proximal rows show the greatest variation in leaflet number, size, and shape. All leaflets have collagenous chordae tendineae arising from the free border and from the parietal side of the leaflets. The endocardium is a flat endothelium which shows a thick, irregular basement membrane. The leaflet body is formed by a loose connective tissue which blends with the subendocardium. The subendocardium is a connective tissue consisting of myofibroblasts, collagen, and elastin. It is divided into two distinct areas: one proximal, which shows little elastin and poorly organized collagen; and one distal, which is rich in elastin, with cells and extracellular fibers organized into layers that are oriented in alternative circumferential and longitudinal directions. The present report is the first systematic analysis of the structure of the sturgeon conus. Descriptions of the conus valves should recognize the existence of three valve rows only. The variability in valve morphology, and the loose structure of the leaflet tissue make it unlikely that the valves play an effective role in preventing blood backflow. In this regard, the ventricle-conus constriction may act as a sphincter. The subendocardium is an elastic coat capable of actively sustaining the tissue deformation that accompanies the heart contractile cycle. Further comparative studies are needed to provide deeper insight into the structural changes that accompany phyletic diversification.


Safety and anticoagulation effect of a low-dose combination of warfarin and aspirin in clinically stable coronary artery disease. Coumadin Aspirin Reinfarction (CARS) Pilot Study Group.
The hypothesis that the combination of low-dose aspirin and warfarin therapy is more effective than aspirin alone in secondary prophylaxis after myocardial infarction is to be examined in the Coumadin Aspirin Reinfarction Study. This pilot study addressed the safety and anticoagulation effect of a fixed, low-dose combination in 114 patients (aged 64 +/- 8 years, 85% men) with stable coronary artery disease receiving 3 mg of warfarin plus 80 mg of aspirin daily for 8 weeks. The international normalized ratio (INR) was measured within 72 hours of initial therapy, and weekly. Of the 110 patients with evaluable INRs, 87 patients (79%) maintained the 3 + 80 mg combination, 19 (17%) had the dose reduced to 1 mg warfarin + 80 mg aspirin, and 4 (4%) discontinued therapy because of a confirmed INR of > or = 4.5. At steady state, patients had INRs of 1.48 +/- 0.41 (3 + 80 mg group) and 1.21 +/- 0.23 (1 + 80 mg group), and inter- and intra-patient variability (estimated by the mean of the between- and within-patient SDs at steady state) was 0.49 +/- 0.08 and 0.13 +/- 0.14, respectively. There was no apparent effect of age on INR distribution. Microscopic hematuria was the most frequent (20%) adverse clinical event, but was unrelated to the INR. Three patients required discontinuation of therapy because of bleeding events (persistent hematuria and epistaxis). A fixed low-dose combination of warfarin and aspirin results in a predictable and stable increase in the INR in a large proportion of patients with coronary artery disease.(ABSTRACT TRUNCATED AT 250 WORDS)


Younger and older adults' associative memory for social information: The role of information importance.
The ability to associate items in memory is critical for social interactions. Older adults show deficits in remembering associative information but can sometimes remember high-value information. In two experiments, younger and older participants studied faces, names, and occupations that were of differing social value. There were no age differences in the recall of important information in Experiment 1, but age differences were present for less important information. In Experiment 2, when younger adults' encoding time was reduced, age differences were largely absent. These findings are considered in light of value-directed strategies when remembering social associative information. (PsycINFO Database Record


Population patch clamp electrophysiology: a breakthrough technology for ion channel screening.
Population patch clamp (PPC) is a novel high throughput planar array electrophysiology technique that allows ionic currents to be recorded from populations of cells under voltage clamp. For the drug discovery pharmacologist, PPC promises greater speed and precision than existing methods for screening compounds at voltage-gated ion channel targets. Moreover, certain constitutively active or slow-ligand gated channels that have hitherto proved challenging to screen with planar array electrophysiology (e.g. SK/IK channels) are now more accessible. In this article we review early findings using PPC and provide a perspective on its likely impact on ion channel drug discovery. To support this, we include some new data on ion channel assay duplexing and on modulator assays, approaches that have thus far not been described.


Lack of effect of inhibitors of DNA synthesis/repair on the ionizing radiation-induced chromosomal damage in G2 stage of ataxia telangiectasia cells.
The relationship between the repair processes occuring at the G2 phase of the cell cycle and cytogenetic damage in ataxia telangiectasia (AT) cells was studied. Lymphoblastoid cells derived from normal, heterozygote AT (HzAT) and three AT patients were exposed to X-rays or fission neutrons and post-treated with inhibitors of DNA synthesis/repair, such as inhibitors of DNA polymerases alpha, delta and epsilon (cytosine arabinoside, ara-C; aphidicolin, APC; buthylphenylen-guanine, BuPdG) or ribonucleotide reductase (hydroxyurea, HU). A strong increase of radiation-induced chromosomal aberrations was observed in normal and HzAT cells post-treated with ara-C, APC and HU, but not in the presence of BuPdG. No enhancing effect was observed in cells derived from AT patients, except for HU post-irradiation treatment. These results suggest that the enzymes that can be inhibited by these agents are not directly involved in the repair of radiation damage induced in G2 cells from AT patients, indicating that probably the AT cells that we used lack the capability to transform the primary DNA lesions into reparable products, or that AT cells might contain a mutated form of DNA polymerase resistant to the inhibitors.


Defence mechanisms: the role of physiology in current and future environmental protection paradigms.
Ecological risk assessments principally rely on simplified metrics of organismal sensitivity that do not consider mechanism or biological traits. As such, they are unable to adequately extrapolate from standard laboratory tests to real-world settings, and largely fail to account for the diversity of organisms and environmental variables that occur in natural environments. However, an understanding of how stressors influence organism health can compensate for these limitations. Mechanistic knowledge can be used to account for species differences in basal biological function and variability in environmental factors, including spatial and temporal changes in the chemical, physical and biological milieu. Consequently, physiological understanding of biological function, and how this is altered by stressor exposure, can facilitate proactive, predictive risk assessment. In this perspective article, existing frameworks that utilize physiological knowledge (e.g. biotic ligand models, adverse outcomes pathways and mechanistic effect models), are outlined, and specific examples of how mechanistic understanding has been used to predict risk are highlighted. Future research approaches and data needs for extending the incorporation of physiological information into ecological risk assessments are discussed. Although the review focuses on chemical toxicants in aquatic systems, physical and biological stressors and terrestrial environments are also briefly considered.


Cell-protective monoclonal antibodies to bovine enterovirus-3 and partial or no activity against other serotypes.
Preparation of monoclonal antibodies to bovine virus diarrhea virus (BVDV) yielded some hybridoma cells that secreted monoclonal antibodies against the Madin-Darby bovine kidney cells. The anti-cellular monoclonal antibodies reacted with other bovine cells (bovine turbinate and testicle) but not with cell lines derived from other animal species. Subclones derived from one hybridoma partially blocked the infectivity of BVDV, possibly through the binding of the monoclonal antibodies with an epitope close to the receptor site of BVDV and not by way of steric hindrance. Unexpectedly, these same subclones completely blocked the infectivity of bovine enterovirus-3 (BEV-3) strain 240A and partially blocked the infectivity of BEV-2 and BEV-3 (ATCC strain) but not that of other serotypes. Other subclones derived from two other hybridomas, although cell membrane specific, did not have a protective activity against BEV or BVDV.


Draft Genome Sequence of a Heterotrophic Facultative Anaerobic Thermophilic Bacterium, Ardenticatena maritima Strain 110ST.
Ardenticatena maritima strain 110S(T) is a filamentous bacterium isolated from an iron-rich coastal hydrothermal field, and it is a unique isolate capable of dissimilatory iron or nitrate reduction among the members of the bacterial phylum Chloroflexi. Here, we report the draft genome sequence comprising 3,569,367 bp, containing 3,355 predicted coding sequences (CDSs).


Assembly of dimeric variants of coumermycins by tandem action of the four biosynthetic enzymes CouL, CouM, CouP, and NovN.
Coumermycin A(1) is a member of the aminocoumarin family of antibiotics. Unlike its structural relatives, novobiocin and clorobiocin, coumermycin A(1) is a dimer built on a 3-methyl-2,4-dicarboxypyrrole scaffold and bears two decorated noviose sugar components which are the putative target binding motifs for DNA gyrase. Starting with this scaffold, we have utilized the ligase CouL for mono- and bisamide formation with aminocoumarins to provide substrates for the glycosyltransferase CouM. CouM was subsequently shown to catalyze mono- and bisnoviosylation of the resulting CouL products. CouP was shown to possess 4'-O-methyltransferase activity on products from tandem CouL, CouM assays. A fourth enzyme, NovN, the 3'-O-carbamoyltransferase from the novobiocin operon, was then able to carbamoylate either or both arms of the CouP product. The tandem action of CouL, CouM, CouP, and NovN thus generates a biscarbamoyl analogue of the pseudodimer coumermycin A(1). Starting from alternative dicarboxy scaffolds, these four enzymes can be utilized in tandem to create additional variants of dimeric aminocoumarin antibiotics.


InP-Bovine Serum Albumin Conjugates as Energy Transfer Probes.
The use of indium phosphide (InP) quantum dots (QDs) as biological fluorophores is limited by the low photoluminescence quantum yield (ϕPL) and the lack of effective bioconjugation strategies. The former issue has been addressed by introducing a strain relaxing intermediate shell such as ZnSe, GaP etc. that significantly enhances the ϕPL of InP. Herein, we present an effective strategy for the conjugation of emissive InP/GaP/ZnS QDs with a commonly used globular protein, namely bovine serum albumin (BSA), which generate colloidally stable QD bioconjugates, labeled as InP-BSA and demonstrate its use as energy transfer probes. The conjugate contains one protein per QD, and the circular dichroism spectra of BSA and InP-BSA exhibit similar fractions of α-helix and β-sheet, reflective of the fact that the secondary structure of the protein is intact on binding. More importantly, the fluorescence polarization studies corroborate the fact that the bound protein can hold a variety of chromophoric acceptors. Upon selectively exciting the InP-BSA component in the presence of bound chromophores, a reduction in the emission intensity of the donor is observed with a concomitant increase in emission of the acceptor. Time-resolved investigations further confirm an efficient nonradiative energy transfer from InP-BSA to the bound acceptors.


Surgical treatment of discrete subaortic stenosis in an adult.
We report on an adult patient with discrete-type subaortic stenosis. A 48-year-old man who had progressed asymptomatically since childhood despite heart murmur was transferred to our hospital. The patient was diagnosed as having severe aortic stenosis with a pressure gradient of 100 mmHg across the aortic valve, associated with a grade II aortic regurgitation. A conventional aortic valve replacement was scheduled. During surgery, the aortic valve was found to be tricuspid but incompetent as a result of shrinking and thickening of the left coronary cusp. A circumferential fibromuscular ridge was observed under the cusps, which corresponded to Kelly's type II discrete subaortic stenosis. Because of the small subaortic area and deformity of the cusp, we performed aortic valve replacement after excision of all cusps and the fibromuscular ridge. Early corrective surgery is recommended for discrete subaortic stenosis to prevention regurgitation progression.


Vitamins and Microelement Bioavailability in Different Stages of Chronic Kidney Disease.
Chronic kidney disease (CKD) predisposes one to either deficiency or toxic excess of different micronutrients. The knowledge on micronutrients-specifically water-soluble vitamins and trace elements-in CKD is very limited. Consequently, current guidelines and recommendations are mostly based on expert opinions or poor-quality evidence. Abnormalities of micronutrient resources in CKD develop for several reasons. Dietary restrictions and anorexia lead to an insufficient micronutrient intake, while diuretics use and renal replacement therapy lead to their excessive losses. Absorption is unpredictable, and metabolism impaired. Better understanding of the micronutrient needs of CKD patients could have an impact on many complications linked to vitamin and trace element disorders, including high mortality, increased risk of atherosclerosis, inflammation, oxidative stress, anemia, polyneuropathy, encephalopathy, weakness and fragility, muscle cramps, bone disease, depression, or insomnia. Here, we summarize the up-to-date knowledge on micronutrient resources in different stages of CKD, and share our experience with the assessment of micronutrient status.


Effectiveness of a virtual vs. in-person group-based education curriculum to increase disease-related knowledge and change health behaviour among cardiac rehabilitation participants.
To investigate the effectiveness of a virtual 12-week group-based education curriculum and to compare results with a retrospective cohort that received the same education in-person for 24 weeks. Participants completed online surveys (pre- and post-intervention) assessing disease-related knowledge, adherence to the Mediterranean diet, exercise self-efficacy, and satisfaction. The number of steps taken per day was recorded using a wearable device. Paired t tests and repeated measures ANOVA were used. A Bonferroni correction was applied(p < 0.01). 80 CR participants receiving virtual education completed both assessments. Following virtual education, participants significantly increased knowledge(p < 0.001), adherence to the Mediterranean diet(p < 0.001) and number of daily steps(p = 0.01). These results were similarly observed in the in-person education group(n = 80), with no significant differences between groups. Virtual education participants decreased their self-efficacy post-intervention(p < 0.001); in contrast, participants of the in-person education increased their exercise self-efficacy(p < 0.001). Overall,31% of virtual and 71% of in-person education participants reported being satisfied with the education delivery format. A virtual group-based education curriculum was effective at improving knowledge and changing behaviour. Similar results were observed in those that received in-person education. Tailoring virtual education interventions to support exercise self-efficacy is warranted. This study strengthens the evidence supporting virtual education in CR.


Atmospheric Hexachlorocyclohexanes in the North Pacific Ocean and the adjacent Arctic region: spatial patterns, chiral signatures, and sea-air exchanges.
During the 2003 Chinese Arctic Research Expedition (CHINARE2003) from the Bohai Sea to the high Arctic (37 degrees N to 80 degrees N) aboard the icebreaker Xuelong (Snow Dragon), air samples were collected for the analysis of hexachlorocyclohexanes (HCHs) in the North Pacific Ocean and adjacent Arctic region. The sigma HCHs (alpha-HCH + gamma-HCH) ranged from 2.3 to 95.1 pg/m3 with the highest levels observed in Far East Asia (32.5 pg/m3), followed by the North Pacific Ocean (17.0 pg/m3) and the Arctic (7.3 pg/ m3). Compared to previous studies in the same areas in 1990s, our measurements were approximately 1 order of magnitude lower. Because of disproportionate chemical reduction and physical fractioning during long-range transport, the ratios of alpha-HCH to gamma-HCH (alpha/gamma-HCH) showed a significant increasing trend from low to high latitudes, suggesting thatthe alpha/gamma-HCH range of 4-7 could not be used to identify sources of technical HCHs especially in remote areas. The ratios of (+)-alpha-HCH to the sum of (+)-alpha-HCH and (-)-alpha-HCH were on average much more biased from 0.5 compared to previous observations in mid-1990s, indicating the exchange of atmospheric alpha-HCH with those in the oceans, where (+)-alpha-HCH was selectively depleted in biological degradation processes. Estimated fugacity ratios based on available data for both alpha-HCH and gamma-HCH further implied their net volatilization from seawater to air in the Arctic Ocean.


Protein kinase A-mediated phosphorylation of cMyBP-C increases proximity of myosin heads to actin in resting myocardium.
Protein kinase A-mediated (PKA) phosphorylation of cardiac myosin binding protein C (cMyBP-C) accelerates the kinetics of cross-bridge cycling and may relieve the tether-like constraint of myosin heads imposed by cMyBP-C. We favor a mechanism in which cMyBP-C modulates cross-bridge cycling kinetics by regulating the proximity and interaction of myosin and actin. To test this idea, we used synchrotron low-angle x-ray diffraction to measure interthick filament lattice spacing and the equatorial intensity ratio, I(11)/I(10), in skinned trabeculae isolated from wild-type and cMyBP-C null (cMyBP-C(-/-)) mice. In wild-type myocardium, PKA treatment appeared to result in radial or azimuthal displacement of cross-bridges away from the thick filaments as indicated by an increase (approximately 50%) in I(11)/I(10) (0.22+/-0.03 versus 0.33+/-0.03). Conversely, PKA treatment did not affect cross-bridge disposition in mice lacking cMyBP-C, because there was no difference in I(11)/I(10) between untreated and PKA-treated cMyBP-C(-/-) myocardium (0.40+/-0.06 versus 0.42+/-0.05). Although lattice spacing did not change after treatment in wild-type (45.68+/-0.84 nm versus 45.64+/-0.64 nm), treatment of cMyBP-C(-/-) myocardium increased lattice spacing (46.80+/-0.92 nm versus 49.61+/-0.59 nm). This result is consistent with the idea that the myofilament lattice expands after PKA phosphorylation of cardiac troponin I, and when present, cMyBP-C, may stabilize the lattice. These data support our hypothesis that tethering of cross-bridges by cMyBP-C is relieved by phosphorylation of PKA sites in cMyBP-C, thereby increasing the proximity of cross-bridges to actin and increasing the probability of interaction with actin on contraction.


Altered blood-brain barrier permeability and its effect on the distribution of Evans blue and sodium fluorescein in the rat brain applied by intracarotid injection.
The aim was to study the blood-brain permeability according to the distribution in the rat brain of Evans blue (EB) and sodium fluorescein (NaFl) administered by an intracarotid injection. Eighteen animals were divided into six groups according to the state of the blood-brain barrier (BBB) at the moment when the dyes were being applied. In the first two groups, the BBB was intact, in groups 3 and 4 the barrier had been opened osmotically prior to the application of the dyes, and in groups 5 and 6 a cellular edema was induced by hyperhydration before administration of the dyes. The intracellular and extracellular distribution of the dyes was studied by fluorescence microscopy. The histological picture thus represented the morphological correlate of the way BBB permeability had been changed before the application of the dyes.


In vivo measurement of oxygen-derived free radicals during reperfusion injury.
By use of an optimized cytochrome c-based biosensor, superoxide radical production was measured continuously in vivo. The aim of this study was the online detection of superoxide concentration during reperfusion after a variable time of ischemia. Measurements were performed by placing the detecting sensor into gastrocnemius muscle tissue. Ischemia was induced by clamping the vena and arteria femoralis. Current response of the sensor was recorded continuously as an equivalent for superoxide concentration. Ischemia times varied from 5 to 120 minutes. The minimum ischemia time to record superoxide production was 10 minutes. By inducing longer periods of ischemia, an increase in superoxide concentration reached its highest levels at 2 hours. Furthermore, the total time of superoxide production after reperfusion depended on the total time of ischemia.


Vascular smooth muscle cell apoptosis induced by "supercooling" and rewarming.
The underlying mechanisms for the reduction in restenosis caused by cryoplasty for peripheral atherosclerotic lesions are not well understood. Because vascular smooth muscle cells (SMCs) are known to play a critical role in restenosis and neointimal hyperplasia, the aim of this study was to determine SMC survival under conditions of "supercooling" and/or rewarming. Bovine aortic SMCs were supercooled to -10 degrees C for 0, 60, or 120 seconds with a custom-designed conduction cooling stage and then rewarmed to 37 degrees C in an incubator for 0, 12, or 24 hours. A terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was used to measure the degree of apoptosis. Activation of Akt (ie, protein kinase B), a key signal protein involved in cell survival, was assessed by Western blot analysis. An increase in apoptotic SMCs was observed with increasing supercooling and rewarming time. Akt was significantly activated at only the most severe condition (120 seconds of supercooling and 24 hours of rewarming), which showed a 2.03-fold increase compared with the group without rewarming. The data suggest that SMC apoptosis occurs with supercooling and rewarming. Protective cell survival mechanisms were activated only late in the rewarming phase. This may partially explain the long-term patency observed with cryoplasty of atherosclerotic peripheral lesions.


Beta 2-microglobulin from serum associates with MHC class I antigens on the surface of cultured cells.
Beta 2-microglobulin (beta 2-m) is a highly conserved polypeptide (12,000 molecular weight; 12K) noncovalently associated with the heavy chain (45-48K) of the major histocompatibility complex (MHC) class I antigens. Its synthesis is required for expression of the HLA-A/B and H-2K/D heavy chains at the cell surface; beta 2-m is also associated with the human cell-surface antigens T6 and M241 isolated from thymocytes. However, on the T leukaemic cell line MOLT-4 some of the T6 antigens contain a different 12K subunit, termed beta t (refs 3, 7, 8). Purified human beta 2-m can exchange partially both with human beta 2-m associated with HLA-antigens, and with mouse beta 2-m associated with murine alloantigens. As MOLT-4 cells were grown in the presence of fetal calf serum (FCS) and as serum is known to contain some free beta 2-m, we examined whether beta t was bovine beta 2-m which had replaced endogenous beta 2-m on the surface of the cell. Here we show both that beta 2-m from FCS or human serum (HuS) used in cell culture can exchange with beta 2-m on the cell surface, and that beta t is in fact bovine beta 2-m.


HIV-1-infected astrocytes and the microglial proteome.
The human immunodeficiency virus (HIV) invades the central nervous system early after viral exposure but causes progressive cognitive, behavior, and motor impairments years later with the onset of immune deficiency. Although in the brain, HIV preferentially replicates productively in cells of mononuclear phagocyte (MP; blood borne macrophage and microglia), astrocytes also can be infected, at low and variable frequency, particularly in patients with encephalitis. Among their many functions, astrocytes network with microglia to provide the first line of defense against microbial infection; however, very little is known about astrocytes' consequences on MP. Here, we addressed this question using co-culture systems of HIV-infected mouse astrocytes and microglia. Pseudotyped vesicular stomatis virus/HIV was used to circumvent the absence of viral receptors and ensure cell genotypic uniformity for studies of intercellular communication. The study demonstrated that infected astrocytes show modest changes in protein elements compared to uninfected cells. In contrast, infected astrocytes induce robust changes in the proteome of HIV-1-infected microglia. Accelerated cell death and redox proteins, among others, were produced in abundance. The observations confirmed the potential of astrocytes to influence the neuropathogenesis of HIV-1 infection by specifically altering the neurotoxic potential of infected microglia and regulating viral maturation.


Local presence of cytomegalovirus and Candida species vs oral lesions in liver and kidney transplant recipients.
Anti-CMV prophylaxis is currently a routine management in patients after organ transplantation. One of the clinical symptoms of CMV infection may be lesions in the oral cavity.The aim of this study was to determine the relationship between CMV infection, occurrence of Candida species and presence of oral mucosal ulceration in transplant recipients. Twenty fi ve patients after kidney or liver transplantation (mean age 13 years +/-4 years), receiving immunosuppression (tacrolimus, cyclosporine or sirolimus), who presented minor and major recurrent aphtous stomatitis (RAS), atypical ulcerations or focal necrosis in the oral cavity were enrolled into the study. Mean duration of post-transplant follow-up was 4 years and 3 months (+/-3 years and 5 months). Clinical dental examination (Pl I, GI), mycological tests and blood tests for CMV infection (specific IgM antibodies and/or pp65 antygenemia) were performed in all patients. Eighteen specimens of oral mucosa were evaluated for CMV presence in situ. CMV infection was confirmed in 13 patients with oral mucosal ulceration (46.43%); which were accompanied by gingivitis (average GI=1.34); in two cases Candida albicans was identified. DNA of HCMV was found in-situ in 5.5% of all biopsies, and in 9% of biopsies of patients with clinical CMV infection; changes did not show the presence of Candida spp. There is a significant correlation between CMV infection and oral lesions; in some cases, CMV may be a direct cause.


MicroRNA-212 negatively regulates starvation induced autophagy in prostate cancer cells by inhibiting SIRT1 and is a modulator of angiogenesis and cellular senescence.
Among a number of non-coding RNAs, role of microRNAs (miRNAs) in cancer cell proliferation, cancer initiation, development and metastasis have been extensively studied and miRNA based therapeutic approaches are being pursued. Prostate cancer (PCa) is a major health concern and several deregulated miRNAs have been described in PCa. miR-212 is differentially modulated in multiple cancers however its function remains elusive. In this study, we found that miR-212 is downregulated in PCa tissues when compared with benign adjacent regions (n = 40). Also, we observed reduced levels of circulatory miR-212 in serum from PCa patients (n = 40) when compared with healthy controls (n = 32). Elucidating the functional role of miR-212, we demonstrate that miR-212 negatively modulates starvation induced autophagy in PCa cells by targeting sirtuin 1 (SIRT1). Overexpression of miR-212 also leads to inhibition of angiogenesis and cellular senescence. In conclusion, our study indicates a functional role of miR-212 in PCa and suggests the development of miR-212 based therapies.


Pleiotropic effects of O-glycosylation in colon cancer.
Changes in the O-glycosylation of proteins have long been associated with the development of cancer, but establishing causal relationships between altered glycosylation and cancer progression remains incomplete. In this study, the authors perform comparative analyses of the cellular phenotypes, transcriptional changes, and alterations in the glycoproteome in colon cancer cells that differentially express one glycosyltransferase. Their results provide a wealth of data on which future studies can be based.


Muscle response to changing neuronal input in the lobster (Panulirus interruptus) stomatogastric system: slow muscle properties can transform rhythmic input into tonic output.
Slow, non-twitch muscles are widespread in lower vertebrates and invertebrates and are often assumed to be primarily involved in posture or slow motor patterns. However, in several preparations, including some well known invertebrate "model" preparations, slow muscles are driven by rapid, rhythmic inputs. The response of slow muscles to such inputs is little understood. We are investigating this issue with a slow stomatogastric muscle (cpv1b) driven by a relatively rapid, rhythmic neural pattern. A simple model suggests that as cycle period decreases, slow muscle contractions show increasing intercontraction temporal summation and at steady state consist of phasic contractions overlying a tonic contracture. We identify five components of these contractions: total, average, tonic, and phasic amplitudes, and percent phasic (phasic amplitude divided by total amplitude). cpv1b muscle contractions induced by spontaneous rhythmic neural input in vitro consist of phasic and tonic components. Nerve stimulation at varying cycle periods and constant duty cycle shows that a tonic component is always present, and at short periods the muscle transforms rhythmic input into almost completely tonic output. Varying spike frequency, spike number, and cycle period show that frequency codes total, average, and tonic amplitudes, number codes phasic amplitude, and period codes percent phasic. These data suggest that tonic contraction may be a property of slow muscles driven by rapid, rhythmic input, and in these cases it is necessary to identify the various contraction components and their neural coding. Furthermore, the parameters that code these components are interdependent, and control of slow muscle contraction is thus likely complex.


[Evaluation of 2 ports video-assisted thoracoscopic surgery (VATS) using endo-close for primary spontaneous pneumothorax].
We aimed to assess the perioperative outcomes of 2 ports video-assisted thoracoscopic surgery(VATS) using Endo-Close (2 ports VATS) in patient for the primary spontaneous pneumothorax(PSP) compared to conventional 3 ports VATS in our hospital. 31 consecutive patients(11;2 ports VATS and 20;3 ports VATS) since 2009 were enrolled in this study. Endo-Close is a device that for pulling the anchoring thread by puncture 1.5 cm incision at the level of the 7th intercostal space( ICS) at the median axillary line( MAL) for camera port, a 1.5 cm incision at the level of the 4th ICS at the anterior axillary line for working port, and 2 mm puncture at the level of the 5th ICS at the MAL using the Endo-Close were placed for this procedure. Compared with 3 ports VATS, equivalent results were obtained by the present procedure;operation time(58.6±18.3 minutes vs 63.0±15.1 minutes, NS), duration of drainage after operation(1.0±0 days vs 1.3±0.5 days, NS), postoperative hospital stay(3.0±1.5 days vs 3.7±1.4 days, NS) significantly.Blood loss was minimal in both cases. In conclusion, 2 port VATS using Endo-Close proved to be feasible method in surgical treatment for PSP.


Accuracy of molecular diagnostics in pemphigus and bullous pemphigoid: comparison of commercial and modified mosaic indirect immunofluorescence tests as well as enzyme-linked immunosorbent assays.
Pemphigus and bullous pemphigoid (BP) are identified by autoantibodies (abs) against desmoglein 1, 3 (DSG1/3) and BP180/BP230, respectively. A novel mosaic to indirect immunofluorescence (IIF) using purified BP180 recombinant proteins spotted on slide and transfected cells expressing BP230, DSG1, DSG3 is available. The commercial (IgG detection) and modified (IgG4 detection) mosaic for indirect immunofluorescence (IIFc - IIF commercial, IIFm - IIF modified) and IgG ELISAs were evaluated in pemphigus and bullous pemphigoid (BP) molecular diagnostics. To compare diagnostic accuracy of commercial (IgG detection) and modified (IgG4 detection) mosaic IIF assay and to examine the diagnostic value of ELISAs in relation to mosaic IIF in routine laboratory diagnostics of pemphigus and BP. Sera from 37 BP and 19 pemphigus patients were studied. Associations between tests were assessed using Fisher's exact test. There are associations between the positive/negative samples detected by IIFc with desmoglein1 (DSG1)/desmoglein3 (DSG3)/BP230 transfected cells and ELISAs and no association between anti-BP180 IgG detection by IIFc and ELISA. IIFm with DSG1 and DSG3 showed both 100% sensitivity and 100% and 78% specificity, respectively, and 100% and 83% positive predictive value in relation to IIFc. IIFm with BP230 had 87% specificity, 55% sensitivity, whereas IIFm with BP180 had a 100% sensitivity and 13% specificity in relation to IIFc. The IIFc with DSG1/DSG3/BP230 transfected cells, excluding BP180 spots, is an alternative method to ELISA in pemphigus/BP diagnostics. IgG4 antibodies, both pathogenically and diagnostically important, are inconsistently detectable with IIFm.


The yeast homolog of H < beta > 58, a mouse gene essential for embryogenesis, performs a role in the delivery of proteins to the vacuole.
The highly conserved and ubiquitously expressed mouse gene H < beta > 58, identified through insertional mutagenesis, has been shown to be essential for early postimplantation development in mouse, but the mechanism by which it acts is unknown (Radice et al. 1991; Lee et al. 1992). We report here the identification of a yeast gene related to the H < beta > 58 gene and provide biochemical and genetic evidence for its function within the cell. The gene, PEP8, plays a role in the delivery of proteins to the vacuole. Disruption of the gene did not affect cell viability. However, the disruptants were shown to have a defect in the processing of the soluble vacuolar proteases but not the membrane vacuolar hydrolases. The processing defect appeared to be a consequence of the inability of the soluble vacuolar hydrolase to reach the vacuole. Although a small amount of the vacuolar precursors was mis-sorted to the extracellular medium, mis-sorting did not appear to be the primary defect in these cells. Pep8p was identified by epitope tagging of the protein. Biochemical fractionation indicated that the protein was peripherally bound to membranes. Immuno-gold electron microscopy indicated that the Pep8p localized to vacuolar membranes. Complementation experiments with the mouse H < beta > 58 cDNA revealed that a Pep8p-H < beta > 58 fusion protein in which the carboxy-terminal 85 amino acids of Pep8p were replaced by the carboxy-terminal 115 amino acids of H < beta > 58 was functional.(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of 2,3-diarylxanthones by electrospray mass spectrometry: gas-phase chemistry versus known antioxidant activity properties.
Xanthones (XH) are a class of heterocyclic compounds widely distributed in nature that hold numerous noteworthy biological and antioxidant activities. Therefore, it is of utmost importance to achieve relevant detailed structural information to understand and assist prediction of their biological properties. The potential relationship between radical-mediated xanthone chemistry in the gas phase and their promising antioxidant activities has not been previously explored. Protonated xanthones XH1-9 were generated in the gas phase by electrospray ionization (ESI) and the main fragmentation pathways of the protonated XH1-9 formed due to collision-induced dissociation (CID) were investigated. In the CID-MS/MS spectra of [M+H](+) ions of XH1, XH2 and XH4 the product ions formed due to H2 O elimination corresponding to the base peak of the spectra. For the remaining six xanthones (XH3, XH5-9), showing the most promising biological profile, the product ion produced with the highest relative abundance (RA) corresponded to the one formed through concomitant loss of H2 O plus CO. Indicative of an inexistent or lower biological activity is the combined loss of CO plus O unique to the CID-MS/MS spectra of XH1, XH2, XH4, and XH5. The product ion formed by loss of 64 Da (concomitant loss of two molecules of H2 O plus CO) is only observed for xanthones containing a catechol unit (XH3 and XH6-9). This product ion has the highest RA for the most potent scavenger of reactive oxygen and nitrogen species XH9 that contains two of these catechol moieties. A strong relationship between some of the biological activities of the studied 2,3-diarylxanthones and their ESI-MS/MS fragmentation spectra was found. The multivariate statistical analysis results suggest that the selected MS features are related to the important biological features. Copyright © 2016 John Wiley & Sons, Ltd.


Transradial approach to treating endovascular cerebral aneurysms: Case series and technical note.
Several benefits have been described over the years of the transradial versus femoral endovascular approach to cardiac interventions. Consequently, its use has become habitual at most centers that perform cardiac catheterizations. This paper details a right transradial approach, incorporating a variety of coils or flow diverters, which can be utilized for the endovascular treatment of different cerebral aneurysms. From 2014 to 2016, we performed 40 endovascular procedures to treat cerebral aneurysms adopting the same right transradial approach. Five aneurysms were treated with flow diverters and 35 were treated with coils. Seven of these aneurisms were asymptomatic, whereas 33 had already ruptured. Satisfactory treatment was achieved in all cases through the same approach in the absence of any complications. A right transradial approach may be satisfactory for the endovascular treatment of different cerebral aneurysms, including aneurysms in either hemisphere. This is the largest series of cerebral aneurysms treated through a transradial approach.


The implication of ocular manifestation of COVID-19 for medical staff and patients - systematic review.
SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2) is a coronavirus that causes COVID-19 (coronavirus disease 2019) with mild to severe respiratory illness. It is a highly contagious disease transmitted through direct or indirect contact with infected people or contaminated surfaces, mainly through respiratory droplets, but other routes are being investigated. It is known that coronaviruses (CoVs) can cause a variety of ocular pathologies in animals, including conjunctivitis, anterior uveitis, retinitis, and optic neuritis, many of which are severe. However, there is no evidence of the SARS-CoV-2 presence in the eye tissue of asymptomatic patients, even if the symptomatic incidence is low. This systematic review presents updated literature on this issue. COVID-19 has now spread throughout the continents and poses a global threat to public health. The risk of rapidly overloading health care systems and causing substantial mortality worldwide is real. On 11 March 2020, the World Health Organization (WHO) announced coronavirus as a global pandemic. Several studies described a few cases with initial ocular symptoms followed by systemic symptoms of the disease. Although the frequency of transmission of SARS-CoV-2 infection through the eye is low, ocular symptoms are not uncommon in COVID-19. In some cases, eye symptoms may be the first signs of illness. This implies the need for hygienic recommendations and use of personal protective equipment (PPE) for medical staff and other services to minimize COVID-19 infection of both health-care workers and patients. A triage for ophthalmic outpatient clinic is mandatory.


Determinants of arterial stiffness in COPD.
Cardiovascular morbidity and mortality is high in patients with chronic obstructive pulmonary disease (COPD) and arterial stiffness is a potentially modifiable risk factor with added predictive value beyond that obtained from traditional risk factors. Arterial stiffness has been the target of pharmacologic and exercise interventions in patients with COPD, but the effects appear limited to those patients with more significant elevations in arterial stiffness. We aimed to identify predictors of increased arterial stiffness in a cohort with moderate to severe COPD. Aortic pulse wave velocity (aPWV) was measured in subjects with moderate to severe COPD enrolled in a multicenter randomized controlled trial. Subjects were categorized into quartiles based on aPWV values and factors affecting high arterial stiffness were assessed. Multivariate models were created to identify independent predictors of high aPWV, and cardiovascular disease (CVD). 153 patients were included. Mean age was 63.2 (SD 8.2) years and mean FEV1 was 55.4 (SD 15.2) % predicted. Compared to the quartile with the lowest aPWV, subjects in the highest quartile were older, had higher systolic blood pressure (SBP), were more likely to be current smokers, and had greater burden of thoracic aortic calcification. On multivariate analyses, age (adjusted OR 1.14, 95%CI 1.05 to 1.25, p = 0.003) and SBP (adjusted OR 1.06, 95% CI 1.02 to 1.09, p = 0.001) were independent predictors of elevated aPWV. Body mass index, therapy with cholesterol lowering medications and coronary calcification were independent predictors of CVD. Elevated arterial stiffness in patients with COPD can be predicted using age, blood pressure and thoracic aortic calcification. This will help identify subjects for enrollment in clinical trials using aPWV for assessing the impact of COPD therapies on CV outcomes.


Developing human-nonhuman chimeras in human stem cell research: ethical issues and boundaries.
The transplantation of adult human neural stem cells into prenatal non-humans offers an avenue for studying human neural cell development without direct use of human embryos. However, such experiments raise significant ethical concerns about mixing human and nonhuman materials in ways that could result in the development of human-nonhuman chimeras. This paper examines four arguments against such research, the moral taboo, species integrity, "unnaturalness," and human dignity arguments, and finds the last plausible. It argues that the transfer of human brain or retinal stem cells to nonhuman embryos would not result in the development of human-nonhuman chimeras that denigrate human dignity, provided such stem cells are dissociated. The article provides guidelines that set ethical boundaries for conducting such research that are consonant with the requirements of human dignity.


Continuities and discontinuities in children and scholarship.
This article introduces a collection of essays on continuity and discontinuity in cognitive development. In his lead essay, J. Kagan (2008) argues that limitations in past research (e.g., on number concepts, physical solidarity, and object permanence) render conclusions about continuity premature. Commentaries respectively (1) argue that longitudinal contexts are essential for interpreting developmental data, (2) illustrate the value of converging measures, (3) identify qualitative change via dynamical systems theory, (4) redirect the focus from states to process, and (5) review epistemological premises of alternative research traditions. Following an overview of the essays, this introductory article discusses how the search for developmental structures, continuity, and process differs between mechanistic-contextualist and organismic-contextualist metatheoretical frameworks, and closes by highlighting continuities in Kagan's scholarship over the past half century.


Seeing Is Believing: Making Wellbeing More Tangible.
Positive Psychology has been instrumental in promoting wellbeing science in the modern era. However, there are still ways in which positive psychology interventions and positive education programmes can be improved to achieve more robust and sustained effects. One suggested method is to make wellbeing more salient and tangible through the use of objective tools that assess the relationship between psychological and physiological wellbeing, and enable wellbeing status and change to be seen. With the addition of an interdisciplinary team, as well as technology-enabled and pedagogically sound learning tools and approaches, the potential for positive outcomes and impact increases exponentially. Monitoring wellbeing progress in this way can provide evidence, motivation and belief in positive psychology and wellbeing interventions. This can lead to engaged learning, sustained benefits and systemic impact. Positive psychology needs to strategically extend on the emerging work in this field to help everyone, including policy makers, notice and value wellbeing.


Hepatitis C virus infection is blocked by HMGB1 released from virus-infected cells.
High-mobility group box 1 (HMGB1), an abundant nuclear protein that triggers host immune responses, is an endogenous danger signal involved in the pathogenesis of various infectious agents. However, its role in hepatitis C virus (HCV) infection is not known. Here, we show that HMGB1 protein is translocated from the nucleus to cytoplasm and subsequently is released into the extracellular milieu by HCV infection. Secreted HMGB1 triggers antiviral responses and blocks HCV infection, a mechanism that may limit HCV propagation in HCV patients. Secreted HMGB1 also may have a role in liver cirrhosis, which is a common comorbidity in HCV patients. Further investigations into the roles of HMGB1 in the diseases caused by HCV infection will shed light on and potentially help prevent these serious and prevalent HCV-related diseases.


Cycloastragenol Attenuates Osteoclastogenesis and Bone Loss by Targeting RANKL-Induced Nrf2/Keap1/ARE, NF-κB, Calcium, and NFATc1 Pathways.
Osteoporosis, which typically affects postmenopausal women, is an osteolytic disease due to over-activation of osteoclasts. However, current drugs targeting osteoclast inhibition face various side effects, making natural compounds with great interest as alternative treatment options. Cycloastragenol (CAG) is a triterpenoid with multiple biological activities. Previously, CAG's activity against aging-related osteoporosis was reported, but the mechanisms of actions for the activities were not understood. This study demonstrated that CAG dose-dependently inhibited osteoclast formation in receptor activator of nuclear factor-κB ligand (RANKL)-stimulated bone marrow macrophage (BMMs). Mechanism studies showed that CAG inhibited NF-κB, calcium, and nuclear factor of activated T cells 1 (NFATc1) pathways. Additionally, CAG also promoted the nuclear factor-erythroid 2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1)/anti-oxidative response element (ARE) pathway that scavenges reactive oxygen species (ROS). Furthermore, CAG was also found to prevent bone loss of postmenopausal osteoporosis (PMO) in a preclinical model of ovariectomized (OVX) mice. Collectively, our research confirms that CAG inhibits the formation and function of osteoclasts by regulating RANKL-induced intracellular signaling pathways, which may represent a promising alternative for the therapy of osteoclast-related disease.


Literature-Based Discovery of Confounding in Observational Clinical Data.
Observational data recorded in the Electronic Health Record (EHR) can help us better understand the effects of therapeutic agents in routine clinical practice. As such data were not collected for research purposes, their reuse for research must compensate for additional information that may bias analyses and lead to faulty conclusions. Confounding is present when factors aside from the given predictor(s) affect the response of interest. However, these additional factors may not be known at the outset. In this paper, we present a scalable literature-based confounding variable discovery method for biomedical research applications with pharmacovigilance as our use case. We hypothesized that statistical models, adjusted with literature-derived confounders, will more accurately identify causative drug-adverse drug event (ADE) relationships. We evaluated our method with a curated reference standard, and found a pattern of improved performance ~ 5% in two out of three models for gastrointestinal bleeding (pre-adjusted Area Under Curve ≥ 0.6).


Combined chemically enhanced primary sedimentation and biofiltration process for low cost municipal wastewater treatment.
The main objective of wastewater treatment is to remove carbon and other nutrients from municipal and industrial effluents in order to protect the environment and human health. Typical wastewater treatment is usually achieved by a combination of physical, chemical and biological methods. In this work, municipal wastewater was depurated using chemically enhanced primary treatment (CEPT) in combination with a pilot-scale trickling filter. Lab scale experiments (Jar-tests) were carried out in order to determine the optimum dosage of chemicals. Selection criteria were the organic load removal efficiency and the low operational cost. Coagulation-flocculation process was conducted through polyaluminium chloride (PAC) and the cationic polyelectrolyte (Zetag 8180) addition. By combining CEPT and trickling filter, tCOD (total Chemical Oxygen Demand), sCOD (soluble Chemical Oxygen Demand), BOD5 (5-day Biochemical Oxygen Demand), NH4+-N, TSS (Total Suspended Solids), VSS (Volatile Suspended Solids) and PO43--P removal efficiencies were estimated to be 89, 82, 93, 60, 96, 96 and 78%, respectively. It is concluded that biological filtration contributed significantly in nutrients removal processes. Moreover, the obtained effluent was low in carbon and rich in nitrogen, which can be applied for restricted irrigation after disinfection, complying with the discharge limits set in the Greek Joint Ministerial Decree 145116/2011.


Monitoring molecules: insights and progress.
In August, 2014, neuroscientists and physical scientists gathered together on the campus of the University of California, Los Angeles to discuss how to monitor molecules in neuroscience. This field has seen significant growth since its inception in the 1970s. Here, the advances in this field are documented, including its advance into understanding the actions that specific neurotransmitters mediate during behavior.


Global Materials Structure Search with Chemically Motivated Coordinates.
Identification of relevant reaction pathways in ever more complex composite materials and nanostructures poses a central challenge to computational materials discovery. Efficient global structure search, tailored to identify chemically relevant intermediates, could provide the necessary first-principles atomistic insight to enable a rational process design. In this work we modify a common feature of global geometry optimization schemes by employing automatically generated collective curvilinear coordinates. The similarity of these coordinates to molecular vibrations enhances the generation of chemically meaningful trial structures for covalently bound systems. In the application to hydrogenated Si clusters, we concomitantly observe a significantly increased efficiency in identifying low-energy structures and exploit it for an extensive sampling of potential products of silicon-cluster soft landing on Si(001) surfaces.


Design and preliminary testing of a visually guided hearing aid.
An approach to hearing aid design is described, and preliminary acoustical and perceptual measurements are reported, in which an acoustic beam-forming microphone array is coupled to an eye-glasses-mounted eye-tracker. This visually guided hearing aid (VGHA)-currently a laboratory-based prototype-senses direction of gaze using the eye tracker and an interface converts those values into control signals that steer the acoustic beam accordingly. Preliminary speech intelligibility measurements with noise and speech maskers revealed near- or better-than normal spatial release from masking with the VGHA. Although not yet a wearable prosthesis, the principle underlying the device is supported by these findings.


Identification of biomarkers for ovarian cancer using strong anion-exchange ProteinChips: potential use in diagnosis and prognosis.
One hundred eighty-four serum samples from patients with ovarian cancer (n = 109), patients with benign tumors (n = 19), and healthy donors (n = 56) were analyzed on strong anion-exchange surfaces using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technology. Univariate and multivariate statistical analyses applied to protein-profiling data obtained from 140 training serum samples identified three biomarker protein panels. The first panel of five candidate protein biomarkers, termed the screening biomarker panel, effectively diagnosed benign and malignant ovarian neoplasia [95.7% sensitivity, 82.6% specificity, 89.2% accuracy, and receiver operating characteristic (ROC) area under the curve of 0.94]. The other two panels, consisting of five and four candidate protein biomarkers each, effectively distinguished between benign and malignant ovarian neoplasia and were therefore referred to as validation biomarker panel I (81.5% sensitivity, 94.9% specificity, 88.2% accuracy, and ROC = 0.94) and validation biomarker panel II (72.8% sensitivity, 94.9% specificity, 83.9% accuracy, and ROC = 0.90). The three ovarian cancer biomarker protein panels correctly diagnosed 41 of the 44 blinded test samples: 21 of 22 malignant ovarian neoplasias [10 of 11 early-stage ovarian cancer (I/II) and 11 of 11 advanced-stage ovarian cancer (III/IV)], 6 of 6 low malignant potential, 5 of the 6 benign tumors, and 9 of 10 normal patient samples. In conclusion, we have discovered three ovarian cancer biomarker protein panels that, when used together, effectively distinguished serum samples from healthy controls and patients with either benign or malignant ovarian neoplasia.


Measuring universal health coverage in reproductive, maternal, newborn and child health: An update of the composite coverage index.
Monitoring universal health coverage in reproductive, maternal and child health requires appropriate indicators for assessing coverage and equity. In 2008, the composite coverage index (CCI)-a weighted average of eight indicators reflecting family planning, antenatal and delivery care, immunizations and management of childhood illnesses-was proposed. In 2017, the CCI formula was revised to update the family planning and diarrhea management indicators. We explored the implications of adding new indicators to the CCI. We analysed nationally representative surveys to investigate how addition of early breastfeeding initiation (EIBF), tetanus toxoid during pregnancy and post-natal care for babies affected CCI levels and the magnitude of wealth-related inequalities. We used Pearson's correlation coefficient to compare different formulations, and the slope index of inequalities [SII] and concentration index [CIX] to assess absolute and relative inequalities, respectively. 47 national surveys since 2010 had data on the eight variables needed for the original and revised formulations, and on EIBF, tetanus vaccine and postnatal care, related to newborn care. The original CCI showed the highest average value (65.5%), which fell to 56.9% when all 11 indicators were included. Correlation coefficients between pairs of all formulations ranged from 0.93 to 0.99. When analysed separately, 10 indicators showed higher coverage with increasing wealth; the exception was EIBF (SII = -2.1; CIX = -0.5). Inequalities decreased when other indicators were added, especially EIBF-the SII fell from 24.8 pp. to 19.2 pp.; CIX from 7.6 to 6.1. The number of countries with data from two or more surveys since 2010 was 30 for the original and revised formulations and 15 when all the 11 indicators were included. Given the growing importance of newborn mortality, it would be desirable to include relevant coverage indicators in the CCI, but this would lead a reduction in data availability, and an underestimation of coverage inequalities. We propose that the 2017 version of the revised CCI should continue to be used.


The successful application of pyrotinib in the treatment of primary trastuzumab-resistant HER-2-positive breast cancer with bilateral axillary lymph node metastasis: a case report.
Breast cancer patients with synchronous contralateral axillary lymph node metastasis (CAM) are very rare, and there is a lack of published treatment guidelines for this kind of patients. We presented a human epidermal growth factor receptor 2 (HER-2)-positive breast cancer case, who was diagnosed as CAM with primary trastuzumab resistance. In this case report, we describes a 47-year-old woman diagnosed with HER-2 positive breast cancer with regional lymph nodes metastasis. However, physical examination identified contralateral axillary lymph nodes, the auxiliary inspection did not assist in determining the nature of the right axillary lymph nodes, and there was no obvious mass in the right breast. Hence, we performed the core needle biopsy on the right axillary lymph node, which revealed the presence of metastatic breast adenocarcinoma. The patient received trastuzumab-based treatment, but this only afforded a progression-free survival of 5 months, owing to primary trastuzumab resistance. She was then successfully treated with pyrotinib, and the outcome was close to clinical complete response (CCR) with a progression-free survival of over 27 months thus far. This case report may help inform clinicians in the diagnosis of breast cancer with CAM and offer the treatment options in HER-2-positive metastasis breast cancer with primary trastuzumab resistance.


Evaluation of the association between embrasure morphology and central papilla recession by radiographic method.
The objective of this study was to evaluate the association between embrasure morphology and central papilla recession. The central papilla was assessed in 372 adults using standardized periapical radiographs of the maxillary central incisors. The following vertical distances were measured: h1, papilla tip to contact point (PT-CP); h2, proximal cementoenamel junction to contact point (pCEJ-CP); h3, bone crest to contact point (BC-CP); and h4, bone crest to papilla tip (BC-PT). Vertical lines were measured along the long axis of the tooth. Horizontal distance was measured as w2 (distance between the two central incisors at the proximal CEJ level, ie, the interdental width). All participants were divided into four groups according to their w2 and h2 measurements: narrow-long (w2 ≤ 2 mm and h2 > 4 mm), narrow-short (w2 ≤ 2 mm and h2 ≤ 4 mm), wide-long (w2 > 2 mm and h2 > 4 mm), and wide-short (w2 > 2 mm and h2 ≤ 4 mm). A statistically significant positive relationship was evident between central papilla recession and age, and a statistically significant negative relationship was evident between age and papilla height in all study groups. The study found that occurrence of central papilla recession is significantly related to age with a wide interdental width and long pCEJCP distance. However, other factors also determine whether the interdental papilla is present or not, and further study of the interaction among these factors is warranted.


[A case of crushing head injury showing bilateral abducens, facial and acoustic nerve palsies].
Most head injuries are due to two basic mechanisms, contact or acceleration. On the other hand, static loading force makes another type of head injury, so called "crushing head injury". In this report, we discussed the mechanism of the crushing head injury and cranial nerve damage of our case. A 35-year-old male was admitted to our hospital suffering from a crushing head injury. In this accident, his head had been crushed between 1000-kg printing machine and truck bed on both temporal regions slowly. He remained fully conscious. On admission 25 minutes after the injury, he showed bilateral sixth-nerve, seventh-nerve palsies, bilateral hearing loss and obvious bleeding from nares and both ears. Gross motor examination was intact. Skull films demonstrated left temporal linear fracture. CT scan showed remarkable pneumocephalus in the basal cistern but no other intracranial lesions. At discharge, two weeks after the trauma, the patient was alert and remained bilateral sixth-nerve, seventh-nerve palsies, and bilateral hearing loss. At the time of 6 months after the initial injury, bilateral abducens nerve palsies and left facial nerve palsy were improved completely. But he demonstrated right slight facial nerve palsy and bilateral moderate hearing loss continuously. In this rare type of injury, the head of the patients had been crushed slowly by the huge power on both temporal regions. This force makes the avulsion of the petrous bone from the foramen lacerun to the outer side of the bone (Russell WR and Schiller F, 1949). This must tend to stretch the sixth nerve and produce abducens nerve injury.(ABSTRACT TRUNCATED AT 250 WORDS)


Promoting Thermodynamic and Kinetic Stabilities of FA-based Perovskite by an in Situ Bilayer Structure.
The commonly employed formamidinium (FA)-containing perovskite solar cells (PSCs) exhibit a severe phase instability problem, thereby limiting their commercial applications. Here, both phase stability and energy efficiency of FA-based PSCs were improved by treating the perovskite surface with pyrrolidinium hydroiodide (PyI) salts, resulting in a 1D perovskite structure (PyPbI3), stacked on the original 3D perovskite. By employing in situ XRD measurements, we revealed that the temperature-dependent phase transition activation barrier was enhanced after forming the 1D/3D structure, resulting in a prolonged transition time by 30-40-fold. From the first-principle calculations, we found the thermodynamic energy difference between two phases reduced from -0.16 to -0.04 eV after the stacking of 1D PyPbI3, offering additional lifetime improvement. Moreover, the champion 1D/3D bilayer PSC exhibits a boosted power conversion efficiency of 19.62%, versus 18.21% of the control. Such 1D/3D bilayer structure may be employed in PSCs to enhance their phase stability and photovoltaic performance.


Stable and Biocompatible Carbon Nanotube Ink Mediated by Silk Protein for Printed Electronics.
Ink-based processes, which enable scalable fabrication of flexible devices based on nanomaterials, are one of the practical approaches for the production of wearable electronics. However, carbon nanotubes (CNTs), which possess great potential for flexible electronics, are facing challenges for use in inks due to their low dispersity in most solvents and suspicious cytotoxicity. Here, a stable and biocompatible CNT ink, which is stabilized by sustainable silk sericin and free from any artificial chemicals, is reported. The ink shows stability up to months, which can be attributed to the formation of sericin-CNT (SSCNT) hybrid through non-covalent interactions. It is demonstrated that the SSCNT ink can be used for fabricating versatile circuits on textile, paper, and plastic films through various techniques. As proofs of concept, electrocardiogram electrodes, breath sensors, and electrochemical sensors for monitoring human health and activity are fabricated, demonstrating the great potential of the SSCNT ink for smart wearables.


COVID-19-induced immune thrombocytopenic purpura; Immunopathogenesis and clinical implications.
Following the outbreak of the COVID-19 pandemic, millions of people around the world have been affected with SARS-CoV-2 infection. In addition to the typical symptoms, thrombotic events, lymphopenia, and thrombocytopenia have been reported in COVID-19 patients. Immune thrombocytopenic purpura (ITP) is one of the thrombotic events that occur in some COVID-19 patients. Hyperinflammation, cytokine storms, and immune dysregulation in some patients are the cause to the main COVID-19 complications such as ALI (acute lung injury), acute respiratory distress syndrome (ARDS), and multiple organ failure. Disruption in the differentiation of T-cells, enhanced differentiation of Th17 and Th1, cell death (pyroptosis), hyper-inflammation and dysfunction of inflammatory neutrophils and macrophages, and hyperactivity of NLRP3-inflammasome are among the important factors that may be the cause to COVID-19-induced ITP. This study aimed to give an overview of the findings on the immunopathogenesis of ITP and COVID-19-induced ITP. Further studies are required to better understand the exact immunopathogenesis and effective treatments for ITP, especially in inflammatory disorders.


Expression of NUP98/TOP1, but not of TOP1/NUP98, in a treatment-related myelodysplastic syndrome with t(10;20;11)(q24;q11;p15).
The t(11;20)(p15;q11) is a rare but recurrent translocation that so far has been described in only four acute myeloid leukemias (AMLs), two treatment-related myelodysplastic syndromes (t-MDSs), and one case of polycythemia vera. Recently, the t(11;20) was shown to result in a fusion of the NUP98 and TOP1 genes, with expression of the NUP98/TOP1 chimera encoded by the der(11)t(11;20), but not of the reciprocal TOP1/NUP98 on the der(20)t(11;20). The genomic breakpoints were subsequently mapped to introns 13 and 7 of NUP98 and TOP1, respectively. We present here a t-MDS with a three-way variant translocation, t(10;20;11)(q24;q11;p15), that generates a der(11)t(11;20) but not a der(20)t(11;20), strongly suggesting that the der(11) harbors the critical genetic rearrangement. Reverse transcriptase-polymerase chain reaction (RT-PCR) revealed a NUP98/TOP1 fusion in which exon 13 of NUP98 was fused in-frame with exon 8 of TOP1. Extra long (XL) genomic PCR and subsequent sequence analyses showed that the breakpoint in NUP98 occurred at nucleotide (nt) 3461 of intron 13, close to a MER (medium reiteration frequency interspersed repetitive element) repeat, and that the breakpoint in TOP1 was at nt 1436 of intron 7, downstream of a MIR (mammalian-wide interspersed repeats) repetitive element. Genomic XL PCR did not amplify the reciprocal TOP1/NUP98, nor was this chimera expressed, as expected from the cytogenetic finding. The present results provide further support for the involvement of the NUP98/TOP1 transcript, but not of the reciprocal one, in the development of MDS/AML. Furthermore, the three cases genomically characterized to date have all been treatment-related and have all harbored breakpoints in intron 13 of NUP98 and intron 7 of TOP1, suggesting that these introns are susceptible to chemotherapy-induced breakage.


Regional anesthesia in coronavirus disease 2019 pandemic.
Coronavirus disease 19 (COVID-19) has presented numerous challenges to healthcare systems worldwide. The virus is highly contagious and infectious since transmission can occur via multiple routes. General measures to prevent viral transmission to patients and healthcare personnel, such as a reduction in clinical load and routine testing, must be coupled with multiple additional safety measures in perioperative services. Regional anesthesia preserves respiratory function and reduces aerosol-generating procedures typically associated with airway management in general anesthesia, making it the first choice for a suspected or confirmed COVID-19 patient in need of surgery with anesthesia. A well thought out regional anesthetic plan, implementation of hygiene and (personal) safety measures are necessary to ensure the best possible outcome for both the patient and the healthcare staff. The present review addresses both practical and recommended measures for performing regional anesthesia in the pandemic, to ensure patient and staff safety, and equipment protection. Further research and evidence-based guidelines are necessary to devise an established standard of care during the current COVID-19 and possible future pandemics.


GENE FLOW, REFUGIA, AND EVOLUTION OF GEOGRAPHIC VARIATION IN THE SONG SPARROW (MELOSPIZA MELODIA).
We surveyed mtDNA restriction-site variation in song sparrows taken from across their continental range. Despite marked geographic variation in size and plumage color, mtDNA variation was not geographically structured. Subspecies were not identifiable by mtDNA analysis. We suggest that postglaciation dispersal scattered mtDNA haplotypes across the continent, explaining the lack of mtDNA geographic patterns. Evolution of size and plumage coloration has probably proceeded faster than mtDNA evolution, leading to the well-structured continental pattern of morphological variation. We suggest that the nonordered geographic distribution of haplotypes reflects the recency of population establishment following completion of range expansion. Dispersal distance was estimated from the mtDNA data at 6.1 km per generation, an order of magnitude greater than that (0.3 km) estimated from demographic data. Island samples were not especially different from continental ones. Rooting the haplotype cladogram with a putative primitive haplotype identified Newfoundland and the Queen Charlotte Islands as potential sites of recent refugia. We question whether study of geographic variation in song sparrows leads to insights concerning speciation.


Reproducibility in urine peptidome profiling using MALDI-TOF.
MALDI-TOF profiling of low molecular weight peptides (peptidome) usage is limited due to the lack of reproducibility from the confounding inferences of sample preparation, data acquisition, and processing. We applied MALDI-TOF analysis to profile urine peptidome with the aims to: (i) compare centrifugal ultrafiltration and dialysis pretreatments, (ii) determine whether using signal LOD (sLOD), together with data normalization, may reduce MALDI-TOF variability. We also investigated the influence of peaks detection on reproducibility. Dialysis allowed to obtain better MALDI-TOF spectra than ultrafiltration. Within the 1000-4000 m/z range, we identified 120 and 129 peaks in intra- and interassay studies, respectively. To estimate the sLOD, serial dilution of pooled urines up to 1/256 were analyzed in triplicate. Six data normalization strategies were investigated-the mean, median, internal standard, relative intensity, TIC, and linear rescaling normalization. Normalization methods alone performed poorly in reducing features variability while when combined to sLOD adjustment showed an overall reduction in features CVs. Applying a feedback signal processing approach, after median normalization and sLOD adjustment, CVs were reduced from 103 to 26% and 113 to 25% for the intra- and interassay, respectively, and spectra became more comparable in terms of data dispersion.


Prospective long term follow-up of a cohort of patients with giant cell arteritis screened for aortic structural damage (aneurysm or dilatation).
Aortic structural damage (ASD) may complicate the course of patients with giant cell arteritis (GCA). However the frequency and outcome of ASD has not been assessed in long term prospective studies. In a previous screening of 54 biopsy proven GCA patients, significant ASD was detected in 12 (22.2%) after a median follow-up of 5.4 years. These patients were periodically evaluated (every 4 years) over a median of 10.3 years (range 4-16.6 years) in order to investigate the development of new ASD and the outcome of previously detected abnormalities. 18 of the 54 patients abandoned the study due to death or other reasons. The remaining 36 patients were subjected to a second screening and 14 to a third screening. 12 (33.3%) of the 36 patients re-screened and 16 (29.6%) of the initial cohort developed ASD, all but one in the thoracic aorta. Aortic diameters at the ascending and descending aorta significantly increased over time. One patient (1.9% of the initial cohort) died from aortic dissection. Surgery was advised in eight (50%) patients with ASD but could only be performed in three patients (37.7%). The development of ASD was not associated with persistence of detectable disease activity. The incidence of ASD is maximal within the first 5 years after diagnosis but continues developing over time, affecting up to 33.3% of individuals after long term follow-up. Once ASD occurs, dilatation increases over time, underlining the need for periodic evaluation. Surgical repair is feasible in about one-third of candidates.


Short telomere length in peripheral blood leukocyte predicts poor prognosis and indicates an immunosuppressive phenotype in gastric cancer patients.
Compelling evidences indicate that relative telomere length (RTL) in peripheral blood leukocytes (PBLs) can predict the clinical outcome of several cancers. However, to date, the prognostic value of leukocyte RTL in gastric cancer (GC) patients has not been explored. In this study, relative telomere length (RTL) in peripheral blood leukocytes (PBLs) was measured using a real-time PCR-based method in a total of 693 GC patients receiving surgical resection. The prognostic value of leukocyte RTL was first explored in the training set (112 patients) using Kaplan-Meier and Cox proportional hazards regression analyses. Then an independent cohort of 581 patients was used as a validation set. To explore potential mechanism, we detected the immunophenotypes of peripheral blood mononuclear cells and plasma concentrations of several cytokines in GC patients. Patients with short RTL showed significantly worse overall survival (OS) and relapse-free survival (RFS) than those with long RTL in all patient sets. Furthermore, leukocyte RTL and TNM stage exhibited a notable joint effect in prognosis prediction. Integration of TNM stage and leukocyte RTL significantly improved the prognosis prediction efficacy for GC. In addition, we found that patients with short RTL had a higher CD4(+) T cell percentage in PBMCs, CD19(+)IL-10(+) Breg percentage in B cells and plasma IL-10 concentration, indicating an enhanced immunosuppressive status with short leukocyte RTL. In conclusion, our study for the first time demonstrates that leukocyte RTL is an independent prognostic marker complementing TNM stage and associated with an immunosuppressive phenotype in the peripheral blood lymphocytes in GC patients.


Formation of N-nitrosamines during the analysis of municipal secondary biological nutrient removal process effluents by US EPA method 521.
US EPA Method 521 employs activated carbon-based solid phase extraction (SPE) cartridges for analyzing N-nitrosamines. The analysis of N-nitrosamines and their chloramine-reactive and ozone-reactive precursors in nitrified municipal secondary effluent revealed the potential for NDMA to form as an artefact during the analysis. As samples passed through the SPE cartridge, the activated carbon mediated the reaction of nitrite with dimethylamine to form NDMA. The reaction was not significant with tertiary amines. Artefactual NDMA formation was important for nitrite concentrations >0.2 mg/L as N in the Biological Nitrogen Removal (BNR) process effluent. However, it is difficult to define a general threshold for nitrite concentrations, because the importance of the reaction also depends on secondary amine concentrations, which are usually poorly characterized. Pre-treatment of samples with sulfamic acid to destroy nitrite mitigated the artefact. This artefact did not affect NDMA analysis in a nitrified effluent from another facility, likely due to low dimethylamine concentrations. This artefact also did not affect the analysis of primary effluent, due to the lack of nitrite, or the analysis of other N-nitrosamines, likely due to the lack of their secondary amine precursors. Because chloramination does not significantly degrade nitrite, this artefact could affect the analysis of chloramine-reactive N-nitrosamine precursors. Because ozonation rapidly degrades nitrite, it should not affect the analysis of ozone-reactive precursors. However, ozonation at 0.8 mg ozone/mg dissolved organic carbon resulted in significant degradation of all N-nitrosamines, even though simultaneous NDMA formation from ozone-reactive precursors resulted in a net increase in NDMA concentration.


Comparison of Non-Coding RNAs in Exosomes and Functional Efficacy of Human Embryonic Stem Cell- versus Induced Pluripotent Stem Cell-Derived Cardiomyocytes.
Both human embryonic stem cell-derived cardiomyocytes (ESC-CMs) and human induced pluripotent stem cell-derived CMs (iPSC-CMs) can serve as unlimited cell sources for cardiac regenerative therapy. However, the functional equivalency between human ESC-CMs and iPSC-CMs for cardiac regenerative therapy has not been demonstrated. Here, we performed a head-to-head comparison of ESC-CMs and iPSC-CMs in their ability to restore cardiac function in a rat myocardial infarction (MI) model as well as their exosomal secretome. Human ESCs and iPSCs were differentiated into CMs using small molecule inhibitors. Fluorescence-activated cell sorting analysis confirmed ∼85% and ∼83% of CMs differentiated from ESCs and iPSCs, respectively, were positive for cardiac troponin T. At a single-cell level, both cell types displayed similar calcium handling and electrophysiological properties, with gene expression comparable with the human fetal heart marked by striated sarcomeres. Sub-acute transplantation of ESC-CMs and iPSC-CMs into nude rats post-MI improved cardiac function, which was associated with increased expression of angiogenic genes in vitro following hypoxia. Profiling of exosomal microRNAs (miRs) and long non-coding RNAs (lncRNAs) revealed that both groups contain an identical repertoire of miRs and lncRNAs, including some that are known to be cardioprotective. We demonstrate that both ESC-CMs and iPSC-CMs can facilitate comparable cardiac repair. This is advantageous because, unlike allogeneic ESC-CMs used in therapy, autologous iPSC-CMs could potentially avoid immune rejection when used for cardiac cell transplantation in the future. Stem Cells 2017;35:2138-2149.


Automated Design Framework for Synthetic Biology Exploiting Pareto Optimality.
In this work we consider Pareto optimality for automated design in synthetic biology. We present a generalized framework based on a mixed-integer dynamic optimization formulation that, given design specifications, allows the computation of Pareto optimal sets of designs, that is, the set of best trade-offs for the metrics of interest. We show how this framework can be used for (i) forward design, that is, finding the Pareto optimal set of synthetic designs for implementation, and (ii) reverse design, that is, analyzing and inferring motifs and/or design principles of gene regulatory networks from the Pareto set of optimal circuits. Finally, we illustrate the capabilities and performance of this framework considering four case studies. In the first problem we consider the forward design of an oscillator. In the remaining problems, we illustrate how to apply the reverse design approach to find motifs for stripe formation, rapid adaption, and fold-change detection, respectively.


Teacher stress and cognitive style.
The aim of the study was to investigate the relationship between the teacher's cognitive style and occupational stress. A sample of 212 teachers in Maltese secondary schools completed a questionnaire surveying both the level of overall stress and the severity of four major areas of job stress. They were also given the Cognitive Styles Analysis which assesses two fundamental dimensions of cognitive style, wholist-analytic and verbal-imagery. For the top and bottom 25 per cent of the sample with extreme style characteristics, self-reported teacher stress was not related to style in a simple manner. However, style interacted significantly with teacher age. With regard to the four major areas of occupational stress, significant variations in the levels of stress were observed with style: whereas Analytics reported greater stress than Wholists for 'pupil misbehaviour' and 'poor working conditions', the converse was true for 'poor staff relations' and 'time pressures'.