DNA methyltransferase inhibitors and the development of epigenetic cancer therapies.
Epimutations, such as the hypermethylation and epigenetic silencing of tumor suppressor genes, play a role in the etiology of human cancers. In contrast to DNA mutations, which are passively inherited through DNA replication, epimutations must be actively maintained because they are reversible. In fact, the reversibility of epimutations by small-molecule inhibitors provides the foundation for the use of such inhibitors in novel cancer therapy strategies. Among the compounds that inhibit epigenetic processes, the most extensively studied are DNA methyltransferase inhibitors. In this review, we examine the literature on DNA methyltransferase inhibitors and discuss the efficacy of such compounds as antitumor agents, as evaluated in phase I-III clinical trials. We also discuss future areas of research, including the development of nonnucleoside inhibitors, the application of novel bioanalytical tools for DNA methylation analysis (which will be important for the clinical application of these compounds by allowing rational approaches to trial design), the need to optimize treatment schedules for maximal biologic effectiveness, and the need to define molecular endpoints so that changes induced by demethylating drugs in patients can be monitored during treatment. Assays for genome-wide and tumor-specific DNA methylation also need to be further developed to establish the pharmacodynamic parameters of DNA methyltransferase inhibitors in patients and to provide rational approaches to maximizing the therapeutic efficacy of these compounds.


Children are not small adults: some differences between pediatric and adult cardiac transplantation.
Cardiac transplantation is a viable therapy for end-stage heart disease in both adults and children. Overall posttransplant survival in the pediatric age group (birth up to 18 years of age) is excellent (greater than 65% at 5 years for all age groups), comparable with the overall survival in the adult transplant recipients. Important differences exist regarding indications, evaluation, surgical technique, and posttransplant management. Indications for transplant in pediatric patients include metabolic and genetic forms of cardiomyopathy and structural congenital heart disease. Evaluation should include a metabolic workup, because potential etiologic factors include mitochondrial disorders, and genetic studies if indicated by phenotypic appearance or family pedigree. Children referred for transplantation with congenital heart disease have often had multiple attempts at palliative surgery, which increase peritransplant surgical risks. Key pediatric issues after transplantation include psychosocial support for the patient and family with regard to school, growth, development, and future expectations.


Half conformally flat gradient Ricci almost solitons.
The local structure of half conformally flat gradient Ricci almost solitons is investigated, showing that they are locally conformally flat in a neighbourhood of any point where the gradient of the potential function is non-null. In opposition, if the gradient of the potential function is null, then the soliton is a steady traceless κ-Einstein soliton and is realized on the cotangent bundle of an affine surface.


High-grade dysplastic adenoma-like mass lesions are not an indication for colectomy in patients with ulcerative colitis.
The management of high-grade dysplasia (HGD) in polypoid lesions in patients with ulcerative colitis (UC) is not well characterized. The purpose of this study was to characterize the clinical course of patients with HGD in adenoma-like dysplasia-associated lesion or masses (DALMs) in the absence of any synchronous flat dysplasia. We hypothesize that colectomy is not warranted in patients who undergo complete excision of adenoma-like DALMs with HGD in UC. Pathology and clinical databases were systematically searched for the presence of dysplastic lesions in inflammatory bowel disease from 1997 to 2004. Patients with UC who had adenoma-like DALMs were identified, and a subset with HGD lesions was defined as our study cohort. A total of 102 patients with UC were identified. Thirty of them (29%) had adenoma-like DALMs without synchronous flat dysplasia; 9 of these patients (30%) had HGD in these lesions. Thirty-two surveillance colonoscopies were performed in this cohort (mean 3.6 colonoscopies/patient). The patients were followed for a mean of 76.5 months (52-99 months). Three out of 9 patients (33%) had colectomy. None of the patients in this cohort was detected to have carcinoma in surveillance biopsies and/or in the resection specimens. Our data suggest that the presence of HGD in DALMs does not warrant colectomy. Continued close observation is suggested in this patient cohort after complete excision of polyps. Further prospective evaluation of this patient population is merited.


Unexpected Photodegradation of a Phosphaketenyl-Substituted Germyliumylidene Borate Complex.
The first zwitterionic borata-bis(NHC)-stabilized phosphaketenyl germyliumylidene [(L2 (O=C=P)Ge:] 2 (L2 =(p-tolyl)2 B[1-(1-adamantyl)-3-yl-2-ylidene]2 ) has been synthesized by salt-metathesis reaction of [L2 (Cl)Ge:] 1 with sodium phosphaethynolate [(dioxane)n NaOCP]. Unexpectedly, its exposure to UV light affords, after reductive elimination of the entire PCO group, the unprecedented [L2 Ge-GeL2 ] complex 3 in 54 % yields bearing the Ge22+ ion with Ge in the oxidation state +1. In addition, the 1,3-digermylium-2,4-diphosphacyclobutadiene [L2 Ge(μ-P)2 GeL2 ] 4 and bis(germyliumylidenyl)-substituted diphosphene [(L2 Ge-P=P-GeL2 )] 5 could also be obtained in moderate yields. The formation of 3-5 and their electronic structures have been elucidated with DFT calculations.


In situ GISAXS study of gold film growth on conducting polymer films.
The growth of a thin gold film on a conducting polymer surface from nucleation to formation of a continuous layer with a thickness of several nanometers is investigated in situ with grazing incidence small-angle X-ray scattering (GISAXS). Time resolution is achieved by performing the experiment in cycles of gold deposition on poly(N-vinylcarbazole) (PVK) and subsequently recording the GISAXS data. The 2D GISAXS patterns are simulated, and morphological parameters of the gold film on PVK such as the cluster size, shape, and correlation distance are extracted. For the quantitative description of the cluster size evolution, scaling laws are applied. The time evolution of the cluster morphology is explained with a growth model, suggesting a cluster growth proceeding in four steps, each dominated by a characteristic kinetic process: nucleation, lateral growth, coarsening, and vertical growth. A very limited amount of 6.5 wt % gold is observed to be incorporated inside a 1.2-nm-thick enrichment layer in the PVK film.


Social work in psychiatric home care: regulations, roles, and realities.
Home care services for people with psychiatric diagnoses are increasing in popularity with consumers, providers, and insurance companies. Traditionally, home care social workers have not provided in-home clinical services because of restrictive regulations and industry culture. This article presents strategies for establishing a clinical role for social workers in a psychiatric home care program that meets Medicare, Joint Commission on Accreditation of Healthcare Organizations, and NASW standards. A discussion of skills requirements, appropriate case assignment, professional role clarifications and differentiations, and marketing strategies is presented.


Improvement of the antigenicity of antirabies vaccine by pooling checked by post-challenge vaccination of guinea-pigs.
The author describes some studies carried out at the Pasteur Institute of Southern India, Coonoor, with the object of developing an antirabies vaccine of uniform potency.It was found that by pooling batches of vaccine from several infected sheep brains a vaccine was produced which was superior in antigenicity (as determined by potency tests in mice) to the NIH (United States National Institutes of Health) Reference Vaccine 155-D as well as to most of the individual batches of vaccine tested. Furthermore, the pooled vaccine conferred a significant degree of protection on guinea-pigs challenged with virulent strains of street virus, even when not administered until an hour after infection.A brief outline is given of the method used for pooling the vaccine.


Treatment of non-traumatic rotator cuff tears: A randomised controlled trial with one-year clinical results.
We have compared three different methods of treating symptomatic non-traumatic tears of the supraspinatus tendon in patients above 55 years of age. A total of 180 shoulders (173 patients) with supraspinatus tendon tears were randomly allocated into one of three groups (each of 60 shoulders); physiotherapy (group 1), acromioplasty and physiotherapy (group 2) and rotator cuff repair, acromioplasty and physiotherapy (group 3). The Constant score was assessed and followed up by an independent observer pre-operatively and at three, six and twelve months after the intervention. Of these, 167 shoulders were available for assessment at one year (follow-up rate of 92.8%). There were 55 shoulders in group 1 (24 in males and 31 in females, mean age 65 years (55 to 79)), 57 in group 2 (29 male and 28 female, mean age 65 years (55 to 79)) and 55 shoulders in group 3 (26 male and 29 female, mean age 65 years (55 to 81)). There were no between-group differences in the Constant score at final follow-up: 74.1 (sd 14.2), 77.2 (sd 13.0) and 77.9 (sd 12.1) in groups 1, 2 and 3, respectively (p = 0.34). The mean change in the Constant score was 17.0, 17.5, and 19.8, respectively (p = 0.34). These results suggest that at one-year follow-up, operative treatment is no better than conservative treatment with regard to non-traumatic supraspinatus tears, and that conservative treatment should be considered as the primary method of treatment for this condition.


Emergence of Chirality from Isotropic Interactions of Three Length Scales.
Chirality is known to play a pivotal role in determining material properties and functionalities. However, it remains a great challenge to understand and control the emergence of chirality and the related enantioselective process particularly when the building components of the system are achiral. Here we explore the generic mechanisms driving the formation of two-dimensional chiral structures in systems characterized by isotropic interactions and three competing length scales. We demonstrate that starting from isotropic and rotationally invariant interactions, a variety of chiral ordered patterns and superlattices with anisotropic but achiral units can self-assemble. The mechanisms for selecting specific states are related to the length-scale coupling and the selection of resonant density wave vectors. Sample phase diagrams and chiral elastic properties are identified. These findings provide a viable route for predicting chiral phases and selecting the desired handedness.


Self-Consistent Stochastic Dynamics for Finite-Size Networks of Spiking Neurons.
Despite the huge number of neurons composing a brain network, ongoing activity of local cell assemblies is intrinsically stochastic. Fluctuations in their instantaneous rate of spike firing ν(t) scale with the size of the assembly and persist in isolated networks, i.e., in the absence of external sources of noise. Although deterministic chaos due to the quenched disorder of the synaptic couplings underlies this seemingly stochastic dynamics, an effective theory for the network dynamics of a finite assembly of spiking neurons is lacking. Here, we fill this gap by extending the so-called population density approach including an activity- and size-dependent stochastic source in the Fokker-Planck equation for the membrane potential density. The finite-size noise embedded in this stochastic partial derivative equation is analytically characterized leading to a self-consistent and nonperturbative description of ν(t) valid for a wide class of spiking neuron networks. Power spectra of ν(t) are found in excellent agreement with those from detailed simulations both in the linear regime and across a synchronization phase transition, when a size-dependent smearing of the critical dynamics emerges.


The role of Kupffer cells in carbon tetrachloride intoxication in mice.
Carbon tetrachloride (CCl(4))-induced acute hepatitis is assumed to involve two phases. The initial phase, initiated within 2 h after CCl(4) administration, involves the generation of reactive oxygen species. The second phase is assumed to start about 8 h subsequent to CCl(4) administration and involves the oxidant-induced activation of Kupffer cells, which release various pro-inflammatory mediators such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). We investigated the role of Kupffer cells during CCl(4) intoxication using Nucling-knockout mice (the KO group), in which the number of Kupffer cells is largely reduced. Plasma alanine transaminase and aspartate transaminase levels demonstrated that the liver necrosis during the second phase was significantly alleviated in the KO group compared with that in the wild-type mice (the WT group). Plasma TNF-α concentrations in the WT group significantly increased 24 h after CCl(4) intoxication, whereas those in the KO group did not significantly increase. Plasma IL-6 levels also significantly increased in the WT group 24 h after CCl(4) administration, but those in the KO group did not increase at any time point. These results indicated that excess reactions of Kupffer cells, once primed by oxidants, were involved in the exacerbation of oxidative stress and liver damage during the second phase of CCl(4) intoxication.


Hierarchical self-assembly of heteronuclear co-ordination networks.
Two ligands L(1) and L(2) have been prepared which contain a chelating pyrazolyl-pyridine group with a pendant aromatic nitrile (in L(1), a benzonitrile; in L(2), a naphthonitrile). These ligands react with Ag(I) salts to give a range of infinite coordination networks or dimeric 'boxes' in which the pyrazolyl-pyridine chelates and the aromatic nitrile groups both participate in coordination to Ag(I) ions. In contrast, L(1) and L(2) form simple mononuclear tris-chelates [ML(3)](2+) with first-row transition metal dications (M = Co, Ni, Zn) in which the aromatic nitrile groups are pendant such that the complexes can be used as 'complex ligands'. The crystal structures of [M(L(2))(3)](BF(4))(2) are based on solely the mer tris-chelate geometry although in solution (1)H NMR spectroscopy reveals a mixture of both fac and mer isomers of the tris-chelates. Reaction of these with Ag(I) ions allows the interaction of the pendant nitrile groups with Ag(i) ions to generate coordination networks based on [ML(3)](2+) cations being crosslinked by Ag(I) ions. In these networks the [ML(3)](2+) cations have solely the fac geometry and lie on threefold rotation axes with all three pendant nitrile groups coordinated to Ag(I) ions which are three-coordinate. {[AgM(L(2))(3)][BF(4)](3)}(infinity) (M = Co, Ni) consist of two interpenetrated (10,3)a nets which have opposite chirality at the [M(L(2))(3)](2+) centres but are not strictly enantiomorphic as the two nets are not crystallographically equivalent. {[AgNi(L(1))(3)](BF(4))(3)}(infinity) in contrast contains two-dimensional sheets which have a (6,3) net structure of hexagonal rings of alternative Ni(II) and Ag(I) centres; although not interpenetrating, two such adjacent (and enantiomorphic) sheets interact with each other via numerous CH...pi interactions between aromatic ligands. Formation of these structures shows that the differential reactivity of the two binding sites in L(1) and L(2) (pyrazolyl-pyridine, and nitrile) can be used to generate mixed-metal coordination networks in a hierarchical, stepwise manner.


JAK-STAT Signaling as a Target for Inflammatory and Autoimmune Diseases: Current and Future Prospects.
The Janus kinase/signal transduction and activator of transcription (JAK-STAT) signaling pathway is implicated in the pathogenesis of inflammatory and autoimmune diseases including rheumatoid arthritis, psoriasis, and inflammatory bowel disease. Many cytokines involved in the pathogenesis of autoimmune and inflammatory diseases use JAKs and STATs to transduce intracellular signals. Mutations in JAK and STAT genes cause a number of immunodeficiency syndromes, and polymorphisms in these genes are associated with autoimmune diseases. The success of small-molecule JAK inhibitors (Jakinibs) in the treatment of rheumatologic disease demonstrates that intracellular signaling pathways can be targeted therapeutically to treat autoimmunity. Tofacitinib, the first rheumatologic Jakinib, is US Food and Drug Administration (FDA) approved for rheumatoid arthritis and is currently under investigation for other autoimmune diseases. Many other Jakinibs are in preclinical development or in various phases of clinical trials. This review describes the JAK-STAT pathway, outlines its role in autoimmunity, and explains the rationale/pre-clinical evidence for targeting JAK-STAT signaling. The safety and clinical efficacy of the Jakinibs are reviewed, starting with the FDA-approved Jakinib tofacitinib, and continuing on to next-generation Jakinibs. Recent and ongoing studies are emphasized, with a focus on emerging indications for JAK inhibition and novel mechanisms of JAK-STAT signaling blockade.


Activities of optical laboratories 1 and 2 at the Photon Factory.
Recent activities at optical laboratories 1 and 2 at the Photon Factory are presented. The activities are (i) construction of optical laboratory 1, (ii) reconstruction of optical laboratory 2, (iii) construction of a visible synchrotron radiation beam extraction mirror made of beryllium, (iv) small-beam-profile measurement by means of the image restoration method, (v) application of the adaptive optical system for correction of distorted wavefronts caused by deformation of the extraction mirror, (vi) construction of a synchrotron radiation interferometer for investigation of the spatial coherency of the visible synchrotron radiation beam and its application to the measurement of vertical beam profile and size.


In-line temperature-compensated vector magnetic field sensor with side-polished fiber.
A novel, to the best of our knowledge, vector magnetic field sensor with temperature compensation is proposed and investigated. The proposed sensor is realized by side polishing a multi-mode optical fiber and adopting the surface plasmon resonance (SPR) effect. The side-polished surface is coated with a magnetic fluid (MF) and polydimethylsiloxane (PDMS) successively along the fiber axis. The as-fabricated sensor can be used not only for magnetic field strength and direction sensing, but also for temperature detection. The achieved magnetic field intensity sensitivities are 1720 pm/mT (90° direction) and -710 pm/mT (0° direction), and the temperature sensitivity is -2070 pm/°C. On top of its temperature compensation ability, the easy fabrication and very high sensitivity of the proposed sensor are attractive features for vector magnetic field sensing applications.


The assessment and care of patients with hepatic encephalopathy.
This article provides an overview of the metabolic brain dysfunction hepatic encephalopathy (HE). HE is caused by severe liver cirrhosis and patients will often be treated in a liver unit, but patients with symptoms of HE may require nursing care anywhere in the healthcare system. Therefore it is beneficial for all nurses to have a basic knowledge of HE and this article explains the symptoms and treatment. Possible differential diagnoses are presented, as well as factors that can trigger episodes of HE. Both patients' and relatives' experiences are examined. Finally, the nurse's role in caring for patients with HE is described, along with the dilemmas and challenges involved.


Genomic Surveillance Elucidates Persistent Wild Poliovirus Transmission During 2013-2015 in Major Reservoir Areas of Pakistan.
Despite tremendous efforts in the fight against polio, Pakistan bears the highest proportion of poliomyelitis cases among the 3 endemic countries including Afghanistan and Nigeria. Apart from insecurity and inaccessibility challenges, the substantial shift of unimmunized children from North Waziristan due to recent military operations was presumed to favor the widespread poliovirus infection in Pakistan. To better understand the current epidemiological situation, we analyzed the virologic data of wild poliovirus type 1 (WPV1) strains detected in Pakistan during 2013-2015. Five genetic clusters (A-E) were identified with at least 5% nucleotide divergence in the viral protein 1 (VP1) coding region. Peshawar, Quetta, and Karachi were found to be the major endemic foci where multiple discrete genetic lineages of WPV1 were detected. Phylogenetic analysis suggests that wild poliovirus strains from endemic regions were genetically distant (with 5%-15% VP1 nucleotide divergence) from those detected in North Waziristan cases, excluding the possibility of a recent progenitor of WPV1 instigating single-source transmission across the country. Orphan lineages detected in Rawalpindi, Lahore, Hyderabad, Sukkur, and Jacobabad revealed silent transmission and the need for vigilant surveillance. Sustenance of analogous genetic lineages over a period of 3 years highlights multiple unimmunized foci present to maintain viral genetic diversity. Our findings are inconsistent with the hypothesis that impoverished populations from North Waziristan serve as a possible determinant of widespread poliomyelitis infection in Pakistan and further emphasize the need to scale-up clinical and environmental surveillance as well as immunization activities.


The Effectiveness of Non-Pharmaceutical Interventions Upon Pregnancy-Related Low Back Pain: A Systematic Review and Meta-Analysis.
Low back pain (LBP) is a very common pathology among pregnant women and various methods are used to reduce the pain. The aim of this study is to conduct an evidence-based systematic review and meta-analysis regarding the effectiveness of the interventions used to reduce low back pain related to pregnancy. Methods and materials: The PEDro database, PubMed, and Cochrane Library were searched from January 2012 until December 2020 as well as the reference lists from identified articles. Studies of any non-pharmaceutical treatment to decrease low back pain were included but only randomized controlled trials were selected. The articles found were screened using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) question. Details about the type of intervention, sample size, outcome measures, results, and statistical significance were extracted from the selected studies. A meta-analysis for pain intensity was conducted and the I2 index as well as x2 test were used to determine the heterogeneity between studies. A random-effects meta-analysis was carried out. The aim was to compare the effectiveness between various methods and the typical care provided on low back pain during pregnancy. Results: From all the articles found in the mentioned databases only 13 studies met the criteria. In these studies, exercise, manipulation, ear acupuncture, Kinesio tape, transcutaneous electrical nerve stimulation (TENS), and neuroemotional technique were the interventions used. In the meta-analysis, six studies with 693 participants were included. The interventions were found to have in total a statistically significant effect on low back pain in comparison with the control group that included the typical care provided to pregnant women (95%CI: 0.08 (0.02,0.31), p<0,01) and they had a high heterogeneity (considerable, Tau² = 2.70; Chi² = 64.11, I² = 91%). Exercise and TENS were determined as more effective than the other types of interventions. TENS and progressive muscle relaxation exercises accompanied by music were found to be the most effective interventions. Although exercise decreased LBP it was not found to have a statistically significant result even though it seems to improve the disability and quality of life of pregnant women. Osteopathic manual treatment (OMT), Kinesio tape, and ear acupuncture affected the lumbar pain intensity but the difference compared to typical care or sham treatment was not statistically significant, while yoga did not improve pregnancy-related LBP. Further research is needed to determine the effectiveness of the interventions mentioned.


Biventricular repair of double outlet right ventricle with noncommitted ventricular septal defect.
Double outlet right ventricle (DORV) with noncommitted ventricular septal defect (VSD) (DORVncVSD) represents the most extreme form of DORV, raising challenging surgical difficulties for biventricular repair. The considerable distance between the VSD and the aorta is primarily because of the very abnormal location of the aorta. The definition of DORVncVSD includes: (1) a VSD distant (greater than aortic diameter) from both arterial valves; (2) both great vessels arising fully from the right ventricle; and (3) a double conus. Double outlet right ventricle with noncommitted ventricular septal defect is a primitive right ventricle, as seen during embryologic development, characterized by the presence of the entire conotruncus from the right ventricle. One surgical technique for repair of DORVncVSD is rerouting of the VSD to the aorta by a long intraventricular tunnel. This technique is limited by the presence of conal tricuspid chordae and by the distance between the tricuspid and pulmonary valves, and is associated with an important risk of subaortic obstruction. Rerouting through the pulmonary artery followed by arterial switch seems a more satisfactory surgical solution. When the VSD is distant from the aorta, it is almost always quite close to the pulmonary artery. Rerouting to the pulmonary artery creates a smaller channel, and its application is not limited by the presence of tricuspid chordae or the tricuspid-to-pulmonary valve distance. However, the arterial switch frequently involves relocating complex coronary arteries.


Cross-linked hydrogels for middle ear packing.
To develop an ideal supportive packing material for ossiculoplasty, tympanoplasty, or other otologic procedures. Several materials, namely, Carbylan-SX (P-C; Sentrx Surgical, Inc., Salt Lake City, UT), Gelfoam (P-GF; Pharmacia & Upjohn, Kalamazoo, MI), and Merogel (P-MG; Medtronics, Inc., Minneapolis, MN), were prepared and then placed into a Hartley guinea pig's (Elm Hill, Chelmsford, MA) middle ear cavities through a large myringotomy incision. The contralateral ear underwent a large myringotomy without packing material being placed. Preoperative and posteroperative auditory brainstem response studies were performed using Intelligent Hearing system software. The animals were examined weekly. Two weeks after packing placement, the animals were killed, and the temporal bones were harvested. Whole temporal bone sectioning was performed to analyze the presence of implant, surrounding inflammation, presence of osteoneogenesis and fibrosis, or adhesions. All the materials, except the P-MG, were easy to place into the middle ear cavity. The P-MG contains woven strands that are difficult to trim into the small sizes needed for placement. The P-MG group had a smaller average amount of implant present compared with the other groups at 2 weeks. The degree of osteoneogenesis was similar among the P-GF, P-C, and P-MG groups. The P-MG and P-C groups contained the lowest amount of fibrosis between the implant and surrounding middle ear structures. This study demonstrates promising results with P-C as a potential supportive packing material for otologic procedures. P-C compares favorably with P-MG and P-GF in a guinea pig model with respect to ease of placement and amount of fibrosis.


MicroRNA exocytosis by large dense-core vesicle fusion.
Neurotransmitters and peptide hormones are secreted into outside the cell by a vesicle fusion process. Although non-coding RNA (ncRNA) that include microRNA (miRNA) regulates gene expression inside the cell where they are transcribed, extracellular miRNA has been recently discovered outside the cells, proposing that miRNA might be released to participate in cell-to-cell communication. Despite its importance of extracellular miRNA, the molecular mechanisms by which miRNA can be stored in vesicles and released by vesicle fusion remain enigmatic. Using next-generation sequencing, vesicle purification techniques, and synthetic neurotransmission, we observe that large dense-core vesicles (LDCVs) contain a variety of miRNAs including miR-375. Furthermore, miRNA exocytosis is mediated by the SNARE complex and accelerated by Ca2+. Our results suggest that miRNA can be a novel neuromodulator that can be stored in vesicles and released by vesicle fusion together with classical neurotransmitters.


[Intraoperative laparoscopic cholangiography -- when is it useful?].
Since the introduction of laparoscopic cholecystectomy (LC), a decrease in the practice of intraoperative cholangiography (IOC) has been reported. Are there actually reasons for carrying on IOC during LC? Depending on the management of common bile duct (CBD) stones treatment a different IOC regime is recommended. If the single-stage laparoscopic extraction of ductal calculi during LC is preferred, routine IOC is generally necessary to detect all CBD stones for desobstruction via ductus cysticus or choledochotomy. When therapeutic splitting is favoured, including two-stage management with endoscopic desobstruction and later LC, routine IOC can be foregone. However, selective practice of IOC can help to reduce the rates of unnecessary preoperative investigations from 40-60 % to 20 % when postoperative endoscopic desobstruction demonstrates similar success rates of about 95 %. Regarding the preventive character of laparoscopic IOC to CBD injuries, a routine investigation should be adopted by institutions with injury rates > 0.4 % and in the learning phase of young surgeons. For all other institutions a selective practice should be recommended when difficult intraoperative conditions render recognition of the anatomical situation more difficult or for identification of dissected non-bleeding ducts near the triangle of Calot.


Access to healthcare services by people with intellectual disabilities: a rural-urban comparison.
Access to healthcare services is poor both for adults with intellectual disabilities and for people living in rural areas. Adults with intellectual disabilities in rural areas may therefore be at a double disadvantage. Representative data were analysed from 39 rural and 633 urban participants from Scotland. Information on demographics, healthcare, and access to services were collected from face-to-face interviews and primary care notes. Both direct comparison and binary logistic regression showed the rural sample to have had significantly more contact with primary (odds ratio = 4.02, 95% CI 1.56-10.35, P = 0.004) and secondary health care (OR = 3.93, 95% CI = 1.81-8.55, P = 0.001). Contact with allied healthcare professionals was not significantly different, except that people in the rural sample were significantly more likely to have had recent contact with a dentist and an optician. Contrary to the original hypothesis, adults with intellectual disabilities living in rural areas were not found to be disadvantaged.


[Mechanical lithotripsy during retrograde cholangiography in choledocholithiasis untreatable by conventional endoscopic sphincterotomy].
Endoscopic papillotomy followed by the removal of stones with the Dormia basket is the treatment of choice in residual choledocolithiasis after cholecystectomy and in choledocolithiasis in patients with a high surgical risk. Nonetheless, the size of the stones may, occasionally, impede removal by this procedure. Mechanical lithotripsy may resolve this clinical situation. The authors present the results of this technique in 40 consecutive patients in whom the elimination of the stones was not possible by conventional methods and thus mechanical lithotripsy was performed. This procedure was found to be effective in 32 out of 40 patients (80%). In 22 cases (55%) lithotripsy was effective in one session performed during the first ERCP, 8 (20%) required two procedures and in 2 (5%) three sessions were necessary. In 9 cases the stones were spontaneously eliminated with lavage using physiologic serum or diluted contrast, in 6 the stones were extracted by lithotripsy, in 16 they were eliminated by a conventional Dormia basket and in the remaining case the Fogarty balloon was used. Lithotripsy could not be carried out in 8 out of the 40 patients (20%) due to inability to pick the stone up in 7 cases and because of the impossibility of breaking the stone up with the lithotripsy basket in the remaining case. No complications directly related to lithotripsy were observed. In conclusion, mechanical lithotripsy in an effective and safe therapeutic alternative in large size choledocolithiasis and can be performed during endoscopy.


A first or dominant immunization. I. Suppression of simultaneous cytolytic T cell responses to unrelated alloantigens.
A first or dominant immunization with one antigen markedly inhibited specific cytolytic T lymphocyte (CTL) responses to a second unrelated alloantigen without suppressing antibody responses to other antigens. Suppression was induced rapidly, became systemic, and could be transferred passively with only serum. Suppression did not result from elimination of cells capable of responding to the second antigen. The mechanisms responsible for this "priority of the first response" may be the same that help protect the fetus during pregnancy, promote renal allograft survival after multiple blood transfusions, and prevent effective CTL-mediated immunity to variants of tumor cells or infectious agents that arise during tumor progression or chronic infections.


[Effect of acupoint-catgut-embedding intervention on type II diabetic rats].
To observe the effect of acupoint-catgut-embedding therapy on blood lipid and insulin levels in type I diabetes mellitus rats;s as to reveal its mechanism underlying improvement of pancreatic functions. Thirty SD rats were divided into normal, model and treatment group. The type I diabetes model was established by feeding the rats with high-fat diet and intraperitodneal injection of streptozotocin (STZ). Catgut-embedment was performed in acupoint "Zhongwan" (CV 12)-"Xiawan" (CV 10), "Yishu"-"Ganshu" (BL 18), etc., once every 20 days, twice altogether. Fasting blood glucose (FBG) content was assayed by glucose oxidase method; glycosylated hemoglobin A 1 c (HbA 1 c) assayed by ELISA, and serum fasting C-peptide(FC-p) deterriined by chemiluminescence method; and the triglyceride (TG), total cholesterol (TC), and serum insulin (INS) levels were detected by enzymatic method, separately. The rat's pancreas tissues were fixed with 4% paraformaldehyde and cut into sections (4 Im in thickness) and stained with H. E. method. After modeling, the levels of FBG, HbA 1 c, INS and FC-p, TG, TC were significantly increased in the model group in comparison with those of the normal group (P<0.05), while INS sensitive. index was apparently decreased in the model group (P<0.05). Following catgut-embedding treatment, changes of the above-mentioned indexes were all remarkably reversed (P<0.05), and H.E. staining showed an improvement of the injured pancreatic tissue and islet cells. Catgut-embedding therapy can reduce fasting blood glucose and insulin resistance, and improve lipid metabolism in type I diabetic rats, which may contribute to its effect in protecting pancreatic islet cells.


Transcriptional Bursting in Gene Expression: Analytical Results for General Stochastic Models.
Gene expression in individual cells is highly variable and sporadic, often resulting in the synthesis of mRNAs and proteins in bursts. Such bursting has important consequences for cell-fate decisions in diverse processes ranging from HIV-1 viral infections to stem-cell differentiation. It is generally assumed that bursts are geometrically distributed and that they arrive according to a Poisson process. On the other hand, recent single-cell experiments provide evidence for complex burst arrival processes, highlighting the need for analysis of more general stochastic models. To address this issue, we invoke a mapping between general stochastic models of gene expression and systems studied in queueing theory to derive exact analytical expressions for the moments associated with mRNA/protein steady-state distributions. These results are then used to derive noise signatures, i.e. explicit conditions based entirely on experimentally measurable quantities, that determine if the burst distributions deviate from the geometric distribution or if burst arrival deviates from a Poisson process. For non-Poisson arrivals, we develop approaches for accurate estimation of burst parameters. The proposed approaches can lead to new insights into transcriptional bursting based on measurements of steady-state mRNA/protein distributions.


Future directions in hyperthermia biology.
Eric Hall pointed out many years ago that in hyperthermia the biology is with us, but the physics is against us. Over the years the basic truth in that statement has not wavered. Our best evidence suggests that temperature distributions are 1-1.5 degrees C too low, on average. However, investigations in biology may lead to sensitization of tumours to hyperthermia, even with the temperatures that are already achievable in the clinic. Biologists are now starting to realize that more studies need to be done in the temperature range between 37 and 42 degrees C, since this is the range that is most often reached in the clinic. For example, Dr Hahn coined the term hyperthermic sensitizer for those drugs that are not cytotoxic at 37 degrees C, but have observable cytotoxicity at elevated temperatures. A new class of hyperthermic sensitizers, azo-dihydrochlorides, decompose in aqueous media to release free radicals. Almost no cytotoxicity of these drugs is observed at 37 degrees C, but the drugs are exquisitely cytotoxic at temperatures of 41-42 degrees C. Identification of additional compounds with such properties is warranted, since systemic cytotoxicity could be avoided. Hahn also suggested that the increase in blood flow that occurs in tumours during heating could be exploited as a means to increase drug delivery. Use of this approach to enhance delivery of monoclonal antibodies has proven fruitful. Since long duration heating combined with low dose-rate irradiation leads to obliteration of the radiation dose-rate effect, this approach may be particularly advantageous with monoclonal antibodies radiolabelled with low LET isotopes.(ABSTRACT TRUNCATED AT 250 WORDS)


First Atroposelective Total Synthesis of Enantiomerically Pure Ancistrocladidine and Ancistrotectorine.
The first regio- and stereoselective total synthesis of the axially chiral 7,3'-coupled naphthylisoquinoline alkaloids ancistrocladidine (1) and ancistrotectorine (2) has been described. Both possess a 7,3'-coupled axis, which before now, was difficult to attain synthetically. Moreover, target 2 has a sensitive relative cis-array of the two methyl groups at C1 and C3 in the tetrahydroisoquinoline part. The key step in the chosen strategy was the construction of the biaryl axis in accordance with the "lactone method": the two molecular halves, which were activated in an "inverse-halogenated" form, were prefixed by an ester bridge, followed by intramolecular coupling, and atroposelective cleavage of the lactone auxiliary bridge delivered the desired biaryl scaffold.


Packaging motor from double-stranded RNA bacteriophage phi12 acts as an obligatory passive conduit during transcription.
Double-stranded RNA viruses sequester their genomes within a protein shell, called the polymerase complex. Translocation of ssRNA into (packaging) and out (transcription) of the polymerase complex are essential steps in the life cycle of the dsRNA bacteriophages of the Cystoviridae family (phi6-phi14). Both processes require a viral molecular motor P4, an NTPase, which bears structural and functional similarities to hexameric helicases. In effect, switching between the packaging and the transcription mode requires the translocation direction of the P4 motor to reverse. However, the mechanism of the reversal remains elusive. Here we characterize the P4 protein from bacteriophage phi12 and exploit its purine nucleotide specificity to delineate P4 role in transcription. The results indicate that while P4 actively translocates RNA during packaging it acts as a passive conduit for RNA export. The directionality switching is accomplished via the regulation of P4 NTPase activity within the polymerase core.


Rh(III)-Catalyzed Direct C-H Cyclization of N-Nitrosoanilines with 1,3-Dicarbonyl Compounds: A Route to Tetrahydrocarbazol-4-ones.
A novel and efficient Rh(III)-catalyzed direct C-H bond tandem annulation of N-nitrosoanilines with 1,3-dicarbonyl compounds through two C-H bond cleavage was developed. This protocol provides a rapid access to a series of valuable tetrahydrocarbazol-4-ones with the feature of readily available starting materials, broad functional group tolerance, and in situ generation of carbene precursors. Importantly, some reaction products exhibited promising antiproliferative activity in cancer cell lines.


Biological activities of new anthracyclines containing fluorine, FAD104 and its metabolites.
From various new anthracyclines containing fluorine in their sugar moieties, 7-O-(2,6-dideoxy-2-fluoro-alpha-L-talopyranosyl)adriamycinone-14-hemi pimerate (FAD104) has been selected for clinical investigation, because of its excellent antitumor activity and solubility. In this paper, the mechanism whereby FAD104 exhibits good antitumor activity in vivo was studied through experiments in vitro in comparison with doxorubicin (DOX). FAD104 had weaker activity than DOX in DNA single and double strand scission in P388 cells and in binding to calf thymus DNA. FAD104 was taken up by P388 cells faster than DOX. The ester linkage in FAD104 was gradually hydrolyzed at neutral pH. FAD104 was metabolized to 7-O-(2,6-dideoxy-2-fluoro-alpha-L-talopyranosyl)adriamycinone (FT-ADM) when incubated with mouse or human serum. In mouse serum the esterase activity was about 100 times higher than in human serum. The product of nonenzymatic and enzymatic esterolysis, FT-ADM, was rapidly taken up by P388 cells and accumulated, reaching at a 9.7-fold higher level than DOX. Thus FAD104 was less active than DOX in itself, but it showed much higher activity through conversion into FT-ADM, due to the action of esterase or to spontaneous and gradual hydrolysis at physiological pH. 7-O-(2,6-Dideoxy-2-fluoro-alpha-L-talopyranosyl)adriamycinol (FT-ADM-OH), another metabolite found in mouse serum had the least biological activity among the fluorine-containing anthracycline glycosides, but its activity was still higher than DOX.


The prebiotic molecules observed in the interstellar gas.
Over 130 molecules have been identified in the interstellar gas and circumstellar shells, the largest among them is a carbon chain with 13 atoms and molecular weight of 147 (twice that of the simplest amino acid glycine). The high reliability of astronomical identifications, as well as the fairly accurate quantitative analysis which can often be achieved, is emphasized. Glycine itself has been claimed, but a recent analysis indicates that few, if any, of the astronomical radio lines attributed to glycine are actually from that molecule. Polycyclic aromatic hydrocarbons (PAHs) have long been proposed as the source of the unidentified infrared bands between 3 and 16 microm, but no single PAH has been identified in space, partly because PAHs generally have weak or non-existent radio spectra. A remarkable exception is the non-planar corannulene molecule (C20H10) that has a strong radio spectrum; in the rich molecular cloud TMC-1, it is found that less than 10-5 of the carbon is contained in this molecule, suggesting that PAHs are not the dominant large molecules in the interstellar gas, as has been claimed. Owing to inherent spectroscopic limitations, determining the structures of the large molecules in space may require capture of the dust grains, which are continually entering the outer Solar System.


Stereoselective synthesis of spiropiperidines as BACE-1 aspartyl protease inhibitors via late stage N-arylation of a 1,8-diazaspiro[4.5]dec-3-en-2-one pharmacophore.
A stereoselective synthesis of spiropiperidine compounds, exemplified by compound 1, was developed, which was based upon the late stage N-arylation of a 1,8-diazaspiro[4.5]dec-3-en-2-one pharmacophore. Previously, compound 1 was prepared in low overall yield from piperidinone 2 via the Strecker reaction. A new route was developed, which employed the stereospecific Corey-Link reaction of an enantiomerically pure trichloromethylcarbinol to give a template compound amenable to late stage N-arylation.


The physiology, pharmacology and therapeutic manipulation of the internal anal sphincter.
Recent research into the physiology and pharmacology of the internal anal sphincter has elucidated the importance of this structure in health and disease. Its pharmacological manipulation for therapeutic gain has focused mainly on agents to reduce internal anal sphincter tone, a 'chemical sphincterotomy' that might heal chronic anal fissure. However, drugs to increase sphincter tone, and augment intermittent and appropriate relaxation are also being evaluated. The initial results with this medical approach to anorectal disease have often been disappointing, failing to match the results achievable with surgery, and many of these drugs have a high rate of side effects in the short term. However, clinical trials have yet to establish the optimum doses, dose intervals and routes of administration for many of these therapies. Furthermore, it is uncertain whether this medical approach should be applied to all patients or just to an as yet undefined subgroup. Certainly, even in the current environment of uncertainty, there is little reason not to try medical manipulation of the internal sphincter as first-line treatment. Surgery remains an option for treatment failures; patients responding to pharmacological manipulation of the internal anal sphincter are spared the long term risks of continence that are inherent in many surgical procedures on the anorectum.


Crystal structures of the side-by-side binding of distamycin to AT-containing DNA octamers d(ICITACIC) and d(ICATATIC).
To understand the recognition interactions between AT-containing alternating DNA and minor groove binding drugs, the crystal structures of the side-by-side binding of two distamycin molecules to the DNA octamers d(ICITACIC)2 and d(ICATATIC)2, referred to here as TA and ATAT, respectively, have been determined at 1.6 A and 2.2 A, respectively. Compared to the previous 2:1 all-IC d(ICICICIC)2-distamycin complex, the substitutions of the I x C base-pairs by the A x T base-pairs enable the interactions of the drug with its natural target to be studied. Both complexes assume side-by-side drug binding, isomorphous to the all IC counterpart in the tetragonal space group P4(1)22 (a = b = 28.03 A, c = 58.04 A and a = b = 27.86 A, c = 58.62 A, respectively). The ATAT complex also crystallized in a new polymorphic monoclinic space group C2 (a = 33.38 A, b = 25.33 A, c = 28.11 A and beta = 120.45 degrees) and was solved at 1.9 A resolution. The structures of the three double drug x DNA complexes are very similar, characterized by systematic hydrogen bonding and van der Waals interactions. Each drug hydrogen bonds with the bases of the proximal DNA strand only and stacks with the sugar moiety, while the side-by-side drugs themselves exhibit pyrrole ring-peptide stacking. The pyrrole-peptide interaction is crucial for the side-by-side binding mode of the distamycin/netropsin family of drugs. The purine-pyrimidine alternation is probably responsible for the striking alternation in the helical and backbone conformations. The structures are conserved between the pure IC complex and the AT substituted complexes but further details of the side-by-side binding to DNA are provided by the 1.6 A resolution structure of TA.


Labeling for Quantitative Comparison of Imaging Measurements in Vitro and in Cells.
Qualitative imaging of biomolecular localization and distribution inside cells has revolutionized cell biology. Most of these powerful techniques require modifications to the target biomolecule. Over the past 10 years, these techniques have been extended to quantitative measurements, from in-cell protein folding rates to complex dissociation constants to RNA lifetimes. Such measurements can be affected even when a target molecule is just mildly perturbed by its labels. Here, the impact of labeling on protein (and RNA) structure, stability, and function in cells is discussed via practical examples from the recent literature. General guidelines for selecting and validating modification sites are provided to bring the best from cell biology and imaging to quantitative biophysical experiments inside cells.


Expression of Cell Wall-Modifying Enzymes in Aspen for Improved Lignocellulose Processing.
Wood is an important source of biomass for materials and chemicals, and a target for genetic engineering of its properties for different applications or for research. Wood properties can be altered by using different enzymes acting on cell wall polymers postsynthetically in cell walls. This approach allows for a precise polymer structure modification thanks to the specificity of enzymes used. Such enzymes can originate from all kinds of organisms, or even be modified in a desired way for novel attributes. Here we present a general strategy for expressing a microbial enzyme in aspen and targeting it to cell wall, using an example of fungal glucuronoyl esterase. We describe methods of vector cloning, plant transformation, transgenic line selection and multiplication, testing for the presence of enzymatic activity in different cell compartments, and finally the method of plant transferring from sterile culture to the greenhouse conditions.


Anaemia among adolescent girls in three districts in Ethiopia.
Adolescence is characterized by rapid growth and development with a significantly increased need for macro and micronutrients. However, there is little empirical evidence on the burden of anaemia among adolescent girls in developing countries such as Ethiopia. This study aims to address this gap by evaluating the magnitude of anaemia with an aim to guide design of intervention modalities to address anaemia among adolescent girls. The study employed a community based cross sectional design. The study was conducted on weekends to capture both in school and out of school adolescent girls. Data was collected from a total 1323 adolescent girls. From each district, we randomly selected villages and ensured that the sampled households had a range geographical spread (lowlands, highlands) within the larger category of rural and urban. We performed anaemia testing using HemoCue B-Haemoglobin analyser. We applied a complex survey data analysis method to estimate the level of anaemia. The hemoglobin level was adjusted for altitude and smoking status. We ran a logistic regression model to evaluate predictors of anaemia. The overall anaemia prevalence ranged from 24 to 38%, with an average rate of 29%. Less than half of the girls heard the term anaemia, and about one third knew the relationship between anaemia and the intake of iron rich foods. The risk of anaemia is higher among adolescent girls in their early adolescence period (10-14 years) (Adjusted Odds Ratio (AOR); 1.98; 95% CI; 1.03, 3.82] and among adolescent girls who lived in moderately food insecure households (AOR 1.48; 95% CI; 1.05-2.09). However, knowing the term "anaemia" was found to be protective against the risk of anaemia. The risk of anaemia was particularly high among adolescent girls in their early age and among those living in food insecure households. The prevalence of anaemia among adolescent girls is a moderate public health problem. According to the WHO set criteria, the districts could be candidates for intermittent iron and Folic acid supplementation program.


New European Clinical Trial Regulation: perception and expectations in Italy.
In July 2012, the European Commission formalized the proposal for a European Clinical Trial Regulation that should replace the European Clinical Trials Directive 2001/20/CE. The new Regulation 536/2014 entered into force in June 2014 and it was expected to be applied not earlier than May 2016. Indeed, at the time, all required central tools are not yet available and new forecasts indicate it will become effective at the end of 2018. The aims of the Regulation are the promotion of higher standards in patient's safety and increasing transparency in Clinical Trials, also by changing the application process. An online survey was conducted among the Italian's Clinical Research Coordinators and Clinical Investigators to examine the perception and knowledge about the upcoming changes in Clinical Trials. A total of 190 Clinical Research Coordinators and 80 Clinical Investigators were surveyed. Clinicians are less aware of the content of the Regulation than Clinical Research Coordinators, who demonstrate an extensive expertise on the topic (84.4%), mainly reached through self-training. The majority of the Investigators (74%) believes that their site's facilities and staff already met all the requirements imposed by the Regulation while Clinical Research Coordinators are less optimistic: 65.2% of them believes that the site staff is not yet fully aware of changes associated to its implementation. The general opinion of the interviewed is that the new Regulation will strongly affect the trial management regardless of their type and phase, and the fulfillment of the imposed requirements represents an opportunity that Italy should not miss to increase its attractiveness to the pharmaceutical market.


Atomic-number (Z)-correlated atomic sizes for deciphering electron microscopic molecular images.
SignificanceAtomic resolution transmission electron microscopy (TEM) has opened up a new era of molecular science by providing atomic video images of dynamic motions of single organic and inorganic molecules. However, the images often look different from the images of molecular models, because these models are designed to visualize the electronic properties of the molecule instead of nuclear electrostatic potentials that are felt by the e-beam in TEM imaging. Here, we propose a molecular model that reproduces TEM images using atomic radii correlated to atomic number (Z). The model serves to provide a priori a useful idea of how a single molecule, molecular assemblies, and thin crystals of organic or inorganic materials look in TEM.


Transcatheter coil embolization of an Arc of Buhler aneurysm.
We report the findings of a patient with an asymptomatic Arc of Buhler (AOB) aneurysm, which was successfully treated by transcatheter coil embolization. An abdominal CT and angiography revealed an intact pancreaticoduodenal artery arcade (PDAA) and an anomalous communication between the SMA and celiac axis, termed an AOB. An aneurysm was observed at the origin of the AOB and treated with a transcatheter embolization using coils. A follow-up CT imaging confirmed the total occlusion of the aneurysm with a patent PDAA. The successful results of this treatment suggest that the endovascular therapy of an AOB aneurysm with a celiac axis occlusion and an intact PDAA is feasible and safe.


Unveiling the complexity of transcription factor networks in hematopoietic stem cells: implications for cell therapy and hematological malignancies.
The functionality and longevity of hematopoietic tissue is ensured by a tightly controlled balance between self-renewal, quiescence, and differentiation of hematopoietic stem cells (HSCs) into the many different blood lineages. Cell fate determination in HSCs is influenced by signals from extrinsic factors (e.g., cytokines, irradiation, reactive oxygen species, O2 concentration) that are translated and integrated by intrinsic factors such as Transcription Factors (TFs) to establish specific gene regulatory programs. TFs also play a central role in the establishment and/or maintenance of hematological malignancies, highlighting the need to understand their functions in multiple contexts. TFs bind to specific DNA sequences and interact with each other to form transcriptional complexes that directly or indirectly control the expression of multiple genes. Over the past decades, significant research efforts have unraveled molecular programs that control HSC function. This, in turn, led to the identification of more than 50 TF proteins that influence HSC fate. However, much remains to be learned about how these proteins interact to form molecular networks in combination with cofactors (e.g. epigenetics factors) and how they control differentiation, expansion, and maintenance of cellular identity. Understanding these processes is critical for future applications particularly in the field of cell therapy, as this would allow for manipulation of cell fate and induction of expansion, differentiation, or reprogramming of HSCs using specific cocktails of TFs. Here, we review recent findings that have unraveled the complexity of molecular networks controlled by TFs in HSCs and point towards possible applications to obtain functional HSCs ex vivo for therapeutic purposes including hematological malignancies. Furthermore, we discuss the challenges and prospects for the derivation and expansion of functional adult HSCs in the near future.


Regulating the secondary metabolism in grape berry using exogenous 24-epibrassinolide for enhanced phenolics content and antioxidant capacity.
The objective of this study is to investigate the influence of application of exogenous 24-epibrassinolide (EBR) on the antioxidant capacity, anthocyanins and phenolics content of Vitis vinifera grape berry (cvs. Yan 73 and Cabernet Sauvignon). The grapevine clusters were sprayed with 0 (control), 0.10, 0.40, or 0.80 mg/l of 24-epibrassinolide during veraison, respectively. The EBR application increased the activities of phenylalanine ammonia-lyase (PAL) and UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT), the content of total phenolics, tannins, flavonoids and anthocyanins, individual anthocyanins and the antioxidant capacity of matured grape skins in both varieties. The application also increased the content of flavonoids and anthocyanins in Yan73 pulp. Compared to the other treatments, the treatment of EBR at 0.40 mg/l had significantly higher level than the control in all above assays. Our results indicated that the exogenous EBR treatment can significantly promote grape ripening and enhance anthocyanins and other phenolics contents and antioxidant capacity in the grape skin.


Total and Added Sugar Intake: Assessment in Eight Latin American Countries.
Non-communicable diseases are growing at an alarming rate in Latin America. We assessed total and added sugar intake in Argentina, Brazil, Chile, Colombia, Costa Rica, Ecuador, Peru, and Venezuela, to verify the adequacy of the World Health Organization's recommendations, considering gender, socioeconomic level (SEL) and age. A total of 9218 non-institutionalized individuals living in urban areas (age range 15-65 years) were assessed in the Latin American Study of Nutrition and Health (ELANS), a multicenter household population-based cross-sectional survey. Socio-demographic data were collected. Total and added sugar intakes were measured using two non-consecutive 24-h dietary recalls. The prevalence of excessive sugar intake was estimated. A large proportion of individuals showed high consumption of total and added sugar intake, which reflected in the high prevalence of excessive sugar intake. With minimal differences across countries, in general, women, individuals with high SEL, and younger people had higher percentages of total energy intake from total and added sugar intake, and of contribution of carbohydrates from total and added sugars. Thus, there is high consumption of total and added sugar intake in the Latin American countries with some peculiarities considering socio-demographic variables, which should be considered in each country's health intervention proposals.


SSHscreen and SSHdb, generic software for microarray based gene discovery: application to the stress response in cowpea.
Suppression subtractive hybridization is a popular technique for gene discovery from non-model organisms without an annotated genome sequence, such as cowpea (Vigna unguiculata (L.) Walp). We aimed to use this method to enrich for genes expressed during drought stress in a drought tolerant cowpea line. However, current methods were inefficient in screening libraries and management of the sequence data, and thus there was a need to develop software tools to facilitate the process. Forward and reverse cDNA libraries enriched for cowpea drought response genes were screened on microarrays, and the R software package SSHscreen 2.0.1 was developed (i) to normalize the data effectively using spike-in control spot normalization, and (ii) to select clones for sequencing based on the calculation of enrichment ratios with associated statistics. Enrichment ratio 3 values for each clone showed that 62% of the forward library and 34% of the reverse library clones were significantly differentially expressed by drought stress (adjusted p value < 0.05). Enrichment ratio 2 calculations showed that > 88% of the clones in both libraries were derived from rare transcripts in the original tester samples, thus supporting the notion that suppression subtractive hybridization enriches for rare transcripts. A set of 118 clones were chosen for sequencing, and drought-induced cowpea genes were identified, the most interesting encoding a late embryogenesis abundant Lea5 protein, a glutathione S-transferase, a thaumatin, a universal stress protein, and a wound induced protein. A lipid transfer protein and several components of photosynthesis were down-regulated by the drought stress. Reverse transcriptase quantitative PCR confirmed the enrichment ratio values for the selected cowpea genes. SSHdb, a web-accessible database, was developed to manage the clone sequences and combine the SSHscreen data with sequence annotations derived from BLAST and Blast2GO. The self-BLAST function within SSHdb grouped redundant clones together and illustrated that the SSHscreen plots are a useful tool for choosing anonymous clones for sequencing, since redundant clones cluster together on the enrichment ratio plots. We developed the SSHscreen-SSHdb software pipeline, which greatly facilitates gene discovery using suppression subtractive hybridization by improving the selection of clones for sequencing after screening the library on a small number of microarrays. Annotation of the sequence information and collaboration was further enhanced through a web-based SSHdb database, and we illustrated this through identification of drought responsive genes from cowpea, which can now be investigated in gene function studies. SSH is a popular and powerful gene discovery tool, and therefore this pipeline will have application for gene discovery in any biological system, particularly non-model organisms. SSHscreen 2.0.1 and a link to SSHdb are available from http://microarray.up.ac.za/SSHscreen.


Identification and characterization of a novel expandable adult stem/progenitor cell population in the human exocrine pancreas.
It is a general opinion that tissue-specific stem cells are present in adult tissues but their specific properties remain elusive. They are rare in tissues and heterogeneous; in addition, their identification and the characterization of their progeny has encountered technical difficulties. In particular, the existence of pancreatic stem cells remains elusive because specific markers for their identification are not available. We established a method for the isolation of a population of stem/progenitor cells from the human exocrine pancreas, and propose it as a model for other human compact organs. We also used markers that identified and finally characterized these cells. Spheroids with self-replicative potential were obtained from all specimens. The isolated population contained a subset of CD34+ CD45- cells and was able to generate, in appropriate conditions, colonies that produce insulin. We obtained evidence that most freshly isolated spheroids, when co-cultured with the c-kit positive neuroblastoma cell line LAN 5, produced a c-kit positive progeny of cells larger in their cytoplasmic content than the original spheroid population, with elongated morphology resembling the neuronal phenotype. We identified a novel predominant functional type of stem/progenitor cell within the human exocrine pancreas, able to generate insulin-producing cells and potentially non-pancreatic cells.


Adjuvant methotrexate in the radiotherapeutic management of advanced tumors of the head and neck.
Oral methotrexate (MTX) was administered to a group of 48 patients with advanced head and neck tumors prior to radiation therapy in a nonrandomized manner. A second group of 75 similar patients were randomized to intravenous (i.v.) MTX plus radiation or radiation alone. Three-year survival rates by life table analysis show no significant statistical difference between i.v. MTX plus radiation or radiation alone. Those treated with oral MTX plus radiation have a statistically significant improved survival. The degree of tumor regression in the oral MTX group was correlated with survival. No similar correlation could be found in the intravenous MTX group. The use of either form of MTX correlated with a lower rate of distant metastasis.


Highlights of the twelfth annual meeting of the National Cancer Research Institute (NCRI), 6-9 November 2016, Liverpool, UK.
The annual meeting of the National Cancer Research Institute (NCRI), held in Liverpool, UK, is a multidisciplinary conference. The meeting generally outlines research trends for the coming year and is aimed at cancer professionals at every level. The most important themes discussed for this conference was that of cancer stem cells. Alongside once again prominence was given to topics of cancer evolution and the role of social prevention programmes like previous years.


Development and validation of an improved HPLC-MS/MS method for comparative pharmacokinetics of penehyclidine hydrochloride following a single intravenous or intramuscular injection.
Penehyclidine hydrochloride (PHC) is an anticholinergic drug with both antimuscarinic and antinicotinic activity. In order to compare the pharmacokinetics of two administration routes (intravenous injection (i.v.) and intramuscular injection (i.m.)) of PHC, an improved High Performance Liquid Chromatography Tandem Mass Spectrometry (HPLC-MS/MS) bioanalytical method was developed for the quantification of PHC in plasma and urine using verapamil as the internal standard (I.S.). Chromatography was performed using a Thermo Hypersil GOLD column (30mm×2.1mm, 3μm), with a gradient elution of 1‰ formic acid-10mmol/L ammonium acetate and acetonitrile at 0.3mL/min. Detection and quantitation were performed by electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the positive ion mode. The most intense [M+H](+) MRM transition of PHC at m/z 316.2→128.3 was used for PHC quantitation, and the transition at m/z 454.6→303.2 was used to monitor I.S. The lower limit of quantification (LLOQ) was 0.05 ng/mL. The intraday precision was <6.71% and the interday precision was <11.69%. The pharmacokinetic parameters of i.v. and i.m. administration routes were as follows (i.v. vs i.m.): t1/2 15.73 vs 17.24h, Tmax 0.06 vs 0.26h, AUC0-t 69.35 vs 67.90hng/mL, AUC0-inf 78.24 vs 79.67hng/mL, Cmax 37.5 vs 9.1ng/mL, Ae0-24h 22.7 vs 25.21μg. There were no significant differences between parameters t1/2 and AUC (P>0.05), but significant differences were observed in Cmax, Tmax and Ae0-24h between the two administration routes (P<0.05). The mean absolute bioavailability of the i.m. administration route was 98.4% (95% confidence interval, 93.4-103.6%). Safety results showed that PHC appeared to be well tolerated in both i.v. and i.m. administration routes and pharmacokinetic results showed that PHC was nearly completely absorbed via i.m. administration route.


Spatial context and top-down strategies in visual search.
Marvin M. Chun and Yuhong Jiang (1998) investigated the role of spatial context on visual search. They used two display conditions. In the Old Display condition, the spatial arrangement of items in the search display was kept constant throughout the experiment. In the New Display condition, the spatial arrangement of items was always novel from trial to trial. The results showed better performance with Old Displays than with New Displays. The authors proposed that repeated spatial context help guiding attention to the target location, thus they termed this effect Contextual Cueing. We present three attempts to reproduce this effect. Experiments 1 and 2 were near exact replications of experiments in Chun and Jiang's report, where we failed to obtain Contextual Cueing. Post-experimental interviews revealed that participants used different search strategies when performing the task: an 'active' strategy (an active effort to find the target), or a 'passive' strategy (intuitive search). In Experiment 3, we manipulated task instructions to bias participants into using active or passive strategies. A robust Contextual Cueing Effect was obtained only in the passive instruction condition.


PKCδ as a regulator for TGFβ1-induced α-SMA production in a murine nonalcoholic steatohepatitis model.
The precise mechanism of TGFβ1 signaling in the progression of non-alcoholic steatohepatitis (NASH) has remained unclear. In particular, a potential regulatory mechanism by which PKCδ affects profibrogenic gene expression had never been explored. In this study, therefore, the role of PKCδ in TGFβ1 mediated α-SMA expression was investigated using NASH model mice. In preparation of the NASH model, male C57BL6/J mice were fed a methionine-choline-deficient (MCD) diet for 3 weeks, after which time they were intraperitoneally injected with lipopolysaccharide (LPS). In addition, Tlr4(Lps-d) (CH3/HeJ) mice were used to demonstrate the TGFβ1 signaling's dependency on TLR4 induction. Liver histology and hepatic hepatitis markers were investigated, and hepatic gene expression levels were determined by real-time PCR. Acute liver injury by LPS injection specifically elevated not only α-SMA expression but also phospho-PKCδ in this model. In contrast, Tlr4(Lps-d) (CH3/HeJ) and blockade of TGFβ1 receptor by SB431542 resulted in a significant reduction of PKCδ activation and α-SMA expression. Moreover, the TGFβ1-induced α-SMA production was significantly reduced by a specific PKCδ inhibitor. These findings suggested that PKCδ plays a critical role in TGFβ1-induced α-SMA production in a NASH model. Thus, this was the first demonstration of the involvement of PKCδ in the regulation of α-SMA expression in NASH liver tissues, and the impaired induction of PKCδ phosphorylation by LPS in a steatohepatitis condition. Interestingly, treatment by PKCδ inhibitor caused dramatic reduction of myofibroblast activation, indicating that PKCδ represents a promising target for treating NASH.


Novel approaches to food allergy.
Food allergies have increased in recent decades. However, they cannot be effectively treated by the current management, which is limited to the identification and avoidance of foods that induce allergies and to the use of medicines for symptoms relief. To meet the medical need of prevention and cure of food allergies, several therapeutic strategies are under investigation. Some newly developed biologics such as anti-IgE antibody and anti-interleukin (IL)-5 antibody directed against significant molecules in the allergic process have shown their potential for the treatment of food allergies. Allergen-specific immunotherapy is the therapy that induces immune tolerance and may reduce the need for conventional medication, severity of allergic symptoms and eliminate hypersensitivity. In this article, clinical studies of immunotherapy via subcutaneous, oral, sublingual, and epicutaneous routes are extensively reviewed for their safety and effectiveness on various food allergies. In addition, to reduce the risk of anaphylaxis and increase toleragenic immunity, many studies are focusing on the modification of traditional allergens used for immunotherapy. Moreover, a Chinese herbal formulation with potential anti-allergic effects is being evaluated for its efficacy in patients with peanut allergy. Although more studies are needed, accumulated data of current studies represent compelling evidence of curative effects of some strategies and give a hope that food allergies are likely to be successfully treated in the future.


Increased Pain Symptomatology Among Females vs. Males With Fragile X-Associated Tremor/Ataxia Syndrome.
Individuals with the fragile X premutation report symptoms of chronic pain from multiple systems, have increased incidence of comorbid conditions where pain is a prominent feature, and pathophysiology that supports disrupted pain regulation, inflammation, and energy imbalance. Less is known about how pain manifests for the subpopulation of carriers that develop the motor and cognitive changes of fragile X-associated tremor and ataxia syndrome (FXTAS), and how pain may differ between men and women. We gathered data collected from 104 males and females with FXTAS related to chronic pain, comorbid conditions related to pain, and medications used for pain control to further explore the types of pain experienced and to better characterize how individuals with the fragile X premutation experience pain sensation across genders. We found that women experience significantly more pain symptoms than men, particularly allodynia (20 vs. 2.0%, p = 0.008), peripheral neuropathy pain (43.9 vs. 25.4%, p = 0.0488), migraine (43.9 vs. 14.5%, p = 0.0008), fibromyalgia (26.8 vs. 0%, p = 0.0071) and back pain (48.5 vs. 23.4%, p = 0.008). We found onset of peripheral neuropathy predicts the onset of ataxia (β = 0.63 ± 0.25, p = 0.019) and tremor (β = 0.56 ± 0.17, p = 0.004) across gender. Women also report significantly more anxiety (82.9 vs. 39.7%, p < 0.001), which has implications for ideal pain treatment. These pain symptoms need to be recognized in the medical history and treated appropriately, with consideration for overlapping comorbidities.


Resting-state brain networks in patients with Parkinson's disease and impulse control disorders.
To investigate intrinsic neural networks connectivity changes in Parkinson's disease (PD) patients with and without impulse control disorders (ICD). Fifteen patients with PD with ICD (ICD+), 15 patients with PD without ICD (ICD-) and 24 age and sex-matched healthy controls (HC) were enrolled in the study. To identify patients with and without ICD and/or punding, we used the Minnesota Impulsive Disorders Interview (MIDI) and a clinical interview based on diagnostic criteria for each symptom. All patients underwent a detailed neuropsychological evaluation. Whole brain structural and functional imaging was performed on a 3T GE MR scanner. Statistical analysis of functional data was completed using BrainVoyager QX software. Voxel-based morphometry (VBM) was used to test whether between-group differences in resting-state connectivity were related to structural abnormalities. The presence of ICD symptoms was associated with an increased connectivity within the salience and default-mode networks, as well as with a decreased connectivity within the central executive network (p < .05 corrected). ICD severity was correlated with both salience and default mode networks connectivity changes only in the ICD+ group. VBM analysis did not reveal any statistically significant differences in local grey matter volume between ICD+ and ICD- patients and between all patients and HC (p < .05. FWE). The presence of a disrupted connectivity within the three core neurocognitive networks may be considered as a potential neural correlate of ICD presence in patients with PD. Our findings provide additional insights into the mechanisms underlying ICD in PD, confirming the crucial role of an abnormal prefrontal-limbic-striatal homeostasis in their development.


Bone healing in an aged murine fracture model is characterized by sustained callus inflammation and decreased cell proliferation.
Geriatric fractures take longer to heal and heal with more complications than those of younger patients; however, the mechanistic basis for this difference in healing is not well understood. To improve this understanding, we investigated cell and molecular differences in fracture healing between 5-month-old (young adult) and 25-month-old (geriatric) mice healing utilizing high-throughput analysis of gene expression. Mice underwent bilateral tibial fractures and fracture calluses were harvested at 5, 10, and 20 days post-fracture (DPF) for analysis. Global gene expression analysis was performed using Affymetrix MoGene 1.0 ST microarrays. After normalization, data were compared using ANOVA and evaluated using Principal Component Analysis (PCA), CTen, heatmap, and Incromaps analysis. PCA and cross-sectional heatmap analysis demonstrated that DPF followed by age had pronounced effects on changes in gene expression. Both un-fractured and 20 DPF aged mice showed increased expression of immune-associated genes (CXCL8, CCL8, and CCL5) and at 10 DPF, aged mice showed increased expression of matrix-associated genes, (Matn1, Ucma, Scube1, Col9a1, and Col9a3). Cten analysis suggested an enrichment of CD8+ cells and macrophages in old mice relative to young adult mice and, conversely, a greater prevalence of mast cells in young adult mice relative to old. Finally, consistent with the PCA data, the classic bone healing pathways of BMP, Indian Hedgehog, Notch and Wnt clustered according to the time post-fracture first and age second. Greater understanding of age-dependent molecular changes with healing will help form a mechanistic basis for therapies to improve patient outcomes. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:149-158, 2018.


Trichinella spiralis paramyosin binds to C8 and C9 and protects the tissue-dwelling nematode from being attacked by host complement.
Paramyosin is a thick myofibrillar protein found exclusively in invertebrates. Evidence suggested that paramyosin from helminths serves not only as a structural protein but also as an immunomodulatory agent. We previously reported that recombinant Trichinella spiralis paramyosin (Ts-Pmy) elicited a partial protective immunity in mice. In this study, the ability of Ts-Pmy to bind host complement components and protect against host complement attack was investigated. In this study, the transcriptional and protein expression levels of Ts-Pmy were determined in T. spiralis newborn larva (NBL), muscle larva (ML) and adult worm developmental stages by RT-PCR and western blot analysis. Expression of Ts-Pmy at the outer membrane was observed in NBL and adult worms using immunogold electron microscopy and immunofluorescence staining. Functional analysis revealed that recombinant Ts-Pmy(rTs-Pmy) strongly bound to complement components C8 and C9 and inhibited the polymerization of C9 during the formation of the membrane attack complex (MAC). rTs-Pmy also inhibited the lysis of rabbit erythrocytes (E(R)) elicited by an alternative pathway-activated complement from guinea pig serum. Inhibition of native Ts-Pmy on the surface of NBL with a specific antiserum reduced larvae viability when under the attack of complement in vitro. In vivo passive transfer of anti-Ts-Pmy antiserum and complement-treated larvae into mice also significantly reduced the number of larvae that developed to ML. These studies suggest that the outer membrane form of T. spiralis paramyosin plays an important role in the evasion of the host complement attack.


Changes in dietary fat and fiber and serum hormone concentrations: nutritional strategies for breast cancer prevention over the life course.
The association between dietary fat intake and breast cancer risk has appeared in a meta-analysis of epidemiologic research, migration studies from countries of low to high risk for breast cancer, and animal experiments. With this background, dietary intervention research aims to reduce fat intake and increase fruit, vegetable, and fiber intake, relying on changes in hormone concentrations as biomarkers for reduction in risk of breast cancer. To date, this dietary intervention research spans the life course and has demonstrated stellar success in some studies but sobering results in others. The purpose of this article is to review the intervention research since a 1999 meta-analysis that reported reduced estradiol levels on a low-fat diet and to explore the lessons learned from intervention research on changes in dietary fat and fiber intake and serum hormone concentrations. Secular trends in obesity and ages at pubertal onset and menarche provide dynamic behavioral, genetic, and developmental challenges to the success of dietary prevention. The goal is to formulate an integrative approach to dietary intervention, taking into consideration ethnic group differences in energy expenditure that modulate weight and hormones influencing breast cancer risk over the life course.


Fracture Burden: What Two and a Half Decades of Dubbo Osteoporosis Epidemiology Study Data Reveal About Clinical Outcomes of Osteoporosis.
This review aims to highlight important clinical findings from the over 25 year-long Dubbo Osteoporosis Epidemiology Study particularly focusing on outcomes post fracture. Every low trauma fracture in the elderly is associated with an increased risk of a subsequent fracture, with a higher risk in men than women. All major or proximal fractures and even minor fractures in the very elderly or minor fractures that are then followed by re-fracture are associated with premature mortality, greatest in the first 5 years post fracture. Having a subsequent fracture further increases this high mortality risk, but if an individual survives the high risk period, their risk returns to that of the background population. Non-hip non-vertebral fractures account for a significant proportion of the premature mortality. Despite an improvement in overall health and population mortality over the years, excess mortality post fracture has not changed in the last 2 decades. All low trauma, fractures in the elderly herald a high risk of poor outcomes, particularly in the first few years post fracture. Early intervention should be initiated.


Introducing prospective national registration of knee osteotomies. A report from the first year in Sweden.
Knee osteotomy is a joint preserving surgery with new techniques and implants introduced during recent years. However the information of its use and outcome is scarce. A national knee osteotomy register was started in Sweden in 2013 from which we report here the information gathered during the first year of registration. All patients having knee osteotomy (distal femur and proximal tibia), primaries and re-operations are intended to be included in the prospective registration. Reporting to the register is based on a paper form including information on the patient (ID, sex, age, American Society of Anesthesiologists classification [ASA], weight and height) surgical date, hospital, diagnosis, pre-operative alignment and grade of osteoarthritis (OA), part and LOT numbers of implants, surgical technique, prophylaxis (antithrombotic and antibiotic) and operating time. During the first year (April 2013 to March 2014), 34 clinics reported 220 primary knee osteotomies (209 proximal tibia and 11 distal femur). We estimate that this represents almost 80% of those performed on the adult population during the period. The majority of the patients were classified as healthy (60% ASA grade 1), were men (66%) and the median age was 51 years (range 19-67). Proximal tibia osteotomy for OA performed with open wedge osteotomy using internal fixation without bone transplantation was most commonly reported. As relatively few patients are being treated with different types of fixation and bone substitution in Sweden as well as the rapid development of techniques and new implants, a nationwide registration of knee osteotomies is relevant.


Expansion and Neofunctionalization of Actinoporin-like Genes in Mediterranean Mussel (Mytilus galloprovincialis).
Pore-forming toxins are an important component of the venom of many animals. Actinoporins are potent cytolysins that were first detected in the venom of sea anemones; however, they are occasionally found in animals other than cnidarians and are expanded in a few predatory gastropods. Here, we report the presence of 27 unique actinoporin-like genes with monophyletic origin in Mytilus galloprovincialis, which we have termed mytiporins. These mytiporins exhibited a remarkable level of molecular diversity and gene presence-absence variation, which warranted further studies aimed at elucidating their functional role. We structurally and functionally characterized mytiporin-1 and found significant differences from the archetypal actinoporin fragaceatoxin C. Mytiporin-1 showed weaker permeabilization activity, no specificity towards sphingomyelin, and weak activity in model lipid systems with negatively charged lipids. In contrast to fragaceatoxin C, which forms octameric pores, functional mytiporin-1 pores on negatively charged lipid membranes were hexameric. Similar hexameric pores were observed for coluporin-26 from Cumia reticulata and a conoporin from Conus andremenezi. This indicates that also other molluscan actinoporin-like proteins differ from fragaceatoxin C. Although the functional role of mytiporins in the context of molluscan physiology remains to be elucidated, the lineage-specific gene family expansion event that characterizes mytiporins indicates that strong selective forces acted on their molecular diversification. Given the tissue distribution of mytiporins, this process may have broadened the taxonomic breadth of their biological targets, which would have important implications for digestive processes or mucosal immunity.


Stochastic thermodynamics of Langevin systems under time-delayed feedback control. II. Nonequilibrium steady-state fluctuations.
This paper is the second in a series devoted to the study of Langevin systems subjected to a continuous time-delayed feedback control. The goal of our previous paper [Phys. Rev. E 91, 042114 (2015)PLEEE81539-375510.1103/PhysRevE.91.042114] was to derive second-law-like inequalities that provide bounds to the average extracted work. Here we study stochastic fluctuations of time-integrated observables such as the heat exchanged with the environment, the extracted work, or the (apparent) entropy production. We use a path-integral formalism and focus on the long-time behavior in the stationary cooling regime, stressing the role of rare events. This is illustrated by a detailed analytical and numerical study of a Langevin harmonic oscillator driven by a linear feedback.


[Study on assay strategy of total tritepenoid saponins in traditional Chinese medicines using total saponins from Akebiae Caulis as an example].
Due to lack of reference substances,the content of triterpenoid saponins in traditional Chinese medicines is usually characterized by colorimetric determination of total saponins. However,the specificity of colorimetric method is poor,and the determination result is not accurate enough. So,in this paper,the content determination method of total triterpenoid saponins was studied by taking Akebiae Caulis saponins as an example. The contents of three main saponin aglycones,including arjunolic acid,hederagenin and oleanolic acid,were determined by HPLC method. Referring to the content determination method of total flavonol glycosides in Ginkgo biloba leaves in the 2015 edition of Chinese Pharmacopoeia,the content of Akebiae Caulis saponins was obtained by multiplying the total content of the three above-mentioned aglycones with conversion coefficient. LC-MS/MS analysis results showed that mutongsaponin C and aponin PJIwere the two main triterpene saponins in Akebiae Caulis,and they shared the same molecular formula. So,the average value of the ratios of the molecular weight between mutongsaponin C and the three aglycones was defined as the conversion coefficient.The three aglycones were separated on an ACE Excel 3 C18-AR column( 4. 6 mm×150 mm,3 μm),and methanol-water( containing0. 04% glacial acetic acid and 0. 02% triethylamine) was used as mobile phase with gradient elution. The detection wavelength was set at 210 nm,and the flow rate was 0. 5 m L·min-1. The results showed that there was a good linearity among the ranges of 1. 053-16. 84,0. 200-3. 200 and 1. 515-24. 24 μg for arjunolic acid,hederagenin and oleanolic acid,respectively. Their average recoveries were97. 90%,97. 50% and 100. 5%,with RSD of 2. 0%,2. 9% and 2. 9%,respectively. The results of methodological investigation met the requirements of content determination. The conversion coefficient was 2. 31. This method is simple and reliable,and can be used for the determination of total triterpenoid saponins in Akebiae Caulis. The assay strategy can be used for the determination of total triterpenoid saponins in other traditional Chinese medicines.


Prediction of Cytochrome P450 Inhibition Using a Deep Learning Approach and Substructure Pattern Recognition.
Cytochrome P450 (CYP) is a family of enzymes that are responsible for about 75% of all metabolic reactions. Among them, CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 participate in the metabolism of most drugs and mediate many adverse drug reactions. Therefore, it is necessary to estimate the chemical inhibition of Cytochrome P450 enzymes in drug discovery and the food industry. In the past few decades, many computational models have been reported, and some provided good performance. However, there are still several issues that should be resolved for these models, such as single isoform, models with unbalanced performance, lack of structural characteristics analysis, and poor availability. In the present study, the deep learning models based on python using the Keras framework and TensorFlow were developed for the chemical inhibition of each CYP isoform. These models were established based on a large data set containing 85715 compounds extracted from the PubChem bioassay database. On external validation, the models provided good AUC values with 0.97, 0.94, 0.94, 0.96, and 0.94 for CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, respectively. The models can be freely accessed on the Web server named CYPi-DNNpredictor (cypi.sapredictor.cn), and the codes for the model were made open source in the Supporting Information. In addition, we also analyzed the structural characteristics of chemicals with CYP450 inhibition and detected the structural alerts (SAs), which should be responsible for the inhibition. The SAs were also made available online, named CYPi-SAdetector (cypisa.sapredictor.cn). The models can be used as a powerful tool for the prediction of CYP450 inhibitors, and the SAs should provide useful information for the mechanisms of Cytochrome P450 inhibition.


[The efficacy of cyclosporine A as salvage therapy for severe active ulcerative colitis refractory to glucocorticoid].
Objective: To clarify the efficacy and safety of cyclosporine A (CsA) as salvage therapy in patients with severe active ulcerative colitis (UC) and refractory to steroids. Methods: A total of 24 severe active UC patients refractory to steroids and hospitalized from 2006 to 2012, were retrospectively enrolled.Data including demographic features, clinical manifestations, laboratory tests and medications were collected. Results: CsA was effective in 15(62.5%) patients, who did not receive colectomy during 12-week administration. This regimen was tolerable in most patients.Twelve (50.0%) patients reported 16 adverse events, but only one patient withdrew CsA due to intolerance.The rates of adverse events in initial intravenous CsA including 4 mg·kg(-1)·d(-1,) 3 mg·kg(-1)·d(-1) and 2 mg·kg(-1)·d(-1) were 2/2, 9/17 and 1/5 respectively.Responders had higher white blood cell count compared with non-responders (P= 0.045). Conclusions: CsA could be an effective alternative regimen to colectomy in severe active UC patients who are refractory to steroids.


[Establishment of a model of hydrogen peroxide-induced injury in pulmonary artery endothelium cells and relevant mechanisms of oxidative stress].
Objective: To establish a hydrogen peroxide (H(2)O(2)) induced injury model of pulmonary artery endothelial cells (PAECs) and explore the molecular mechanisms of oxidative stress on the structure and function of PAECs in this model. Methods: Human PAECs were treated with H(2)O(2) at different concentrations (25, 50, 100, 200, 400, 800, 1 600, 3 200, 6 400 μmol/L) for 4 and 24 h, respectively. The PAECs survival curve was obtained according to the cell viability measured by CCK-8 assay. The cell apoptosis of PAECs was detected by flow cytometry. The reactive oxygen species (ROS) generation and mitochondrial activity were measured using small molecule fluorescent probes. Proteins were extracted and the phosphorylation levels of signal molecules in PAECs were detected by Western blot assays. Results: (1) The effect of H(2)O(2) at various concentrations on cell viability of PAECs: cell viability of PAECs decreased in proportion to increasing concentration of H(2)O(2) after incubation for 4 h. The half maximal inhibitory concentration (IC(50)) of PAECs exposed to H(2)O(2) for 4 and 24 h were 397.00 and 488.77 μmol/L, respectively. (2) The effect of H(2)O(2) on cell apoptosis of PAECs: After H(2)O(2) incubation for 4 h, proportions of PAECs at late-apoptosis ((22.58±3.69) %) and necrotic stage( (11.86±4.27)%) were significantly higher than those of control PAECs at late-apoptosis stage( (3.41±1.44)%, P<0.01) and at necrotic stage ((1.94±1.15) % , P<0.05). The survival rate of PAECs post H(2)O(2) was dramatically lower than that of control PAECs ((7.98±3.21)% vs. (48.89±8.08)%, P<0.01). However, there is no statistical difference between both groups regarding to the early apoptosis. (3) The effect of H(2)O(2) on mitochondrial activity and ROS production of PAECs: the mitochondrial activity and ROS generation of PAECs treated by H(2)O(2) were significantly increased compared to those in control PAECs (P<0.01). (4) The effect of H(2)O(2) on signaling molecules in PAECs: there was a significant increase in phosphorylation level of Akt in PAECs incubated with H(2)O(2) for 30 minutes compared to that in control PAECs (P<0.01), while there was no significant difference in levels of Akt between H(2)O(2) treated PAECs and control PAECs. Phosphorylation level of JNK as well as p38 were also significantly upregulated in H(2)O(2) treated PAECs (P<0.01). Conclusion: H(2)O(2) at the concentration of 400 μmol/L could induce human PAECs injuries via the regulation of Akt and MAPK signaling pathways.


What Data Do States Collect Related to School Nurses, School Health, and the Health Care Provided?
School nurses collect data to report to their school district and state agencies. However, there is no national requirement or standard to collect specific data, and each state determines its own set of questions. This study resulted from a joint resolution between the National Association of State School Nurse Consultants and the National Association of School Nurses. The study sought to determine whether similar data points were collected so that comparisons could be made among states and to develop a framework to incorporate the data. Thirty-two states provided their questionnaires or reports. There were 855 data points that could be divided into data related to staff and to students. No categories were measured by all states. The most common data points were the number of students, health screenings, and the number of students with particular conditions for whom the district provided services. A framework for data collection is proposed.


Nucleobase-directed amyloid nanotube assembly.
Cytosine nucleobases were successfully incorporated into the side chain of the self-assembling amyloid peptide fragment HHQALVFFA to give ccAQLVFFA. At a pH range of 3-4, where cytosine is expected to be partially protonated, small-angle X-ray scattering analyses revealed the nucleobase peptide assembles to be well-defined nanotubes with an outer diameter of 24.8 nm and wall thicknesses of 3.3 nm. FT-IR and X-ray diffraction confirmed beta-sheet-rich assembly with the characteristic cross-beta architecture of amyloid. The beta-sheet registry, determined by measuring (13)CO-(13)CO backbone distances with solid-state NMR and linear dichroism, placed the cytosine bases roughly perpendicular to the nanotube axis, resulting in a model where the complementary interactions between the cytosine bases increases beta-sheet stacking to give the nanotube architecture. These scaffolds then extend the templates used to encode biological information beyond the nucleic acid duplexes and into covalent networks whose self-assembly is still defined by a precise complementarity of the side-chain registry.