Identification of human herpesvirus 6 latency-associated transcripts.
Four kinds of latency-associated transcripts of human herpesvirus 6 were identified which were detected only in latently infected cells. Although they were oriented in the same direction as the immediate-early 1 and 2 (IE1/IE2) genes and shared their protein-coding region with IE1/IE2, their transcription start sites and exon(s) were latency associated.


Physical activity, body composition and physical self-esteem: a 3-year follow-up study among adolescents in Sweden.
To measure physical activity by means of daily pedometer steps, body composition, expressed as body mass index (BMI) and bioelectrical impedance as percent body fat, and perceived physical self-esteem and additionally, to evaluate eventual predictors for a healthy lifestyle i.e., highly physically active, normal weighted and a high physical self-esteem in a follow-up group. During autumn 2000 physical activity level was assessed and BMI was calculated in 871 children aged 7-14 in south eastern Sweden and in 501 of these, physical self-esteem were also assessed. During autumn 2003 a total of 375 adolescents aged 15-18 were assessed using the same methodology and additionally percent body fat was measured. Ninety-three of these adolescents (46 girls), were also measured in 2000 and they formed the follow-up group. In the follow-up group a significant increase in BMI and stability in physical self-esteem in boys and girls, and in boys a significant decrease in daily steps was found 3 years later. Strongest predictor to be highly physically active, maintain normal BMI and high self-esteem 3 years later, i.e., to have a healthy lifestyle, was for girls increased self-esteem and for boys a decreased BMI. Consequently, for a healthy lifestyle, promoting physical self-esteem in girls and weight control in boys is of great importance in early adolescence.


Vascular bioactivation of nitroglycerin is catalyzed by cytosolic aldehyde dehydrogenase-2.
According to general view, aldehyde dehydrogenase-2 (ALDH2) catalyzes the high-affinity pathway of vascular nitroglycerin (GTN) bioactivation in smooth muscle mitochondria. Despite having wide implications to GTN pharmacology and raising many questions that are still unresolved, mitochondrial bioactivation of GTN in blood vessels is still lacking experimental support. In the present study, we investigated whether bioactivation of GTN is affected by the subcellular localization of ALDH2 using immortalized ALDH2-deficient aortic smooth muscle cells and mouse aortas with selective overexpression of the enzyme in either cytosol or mitochondria. Quantitative Western blotting revealed that ALDH2 is mainly cytosolic in mouse aorta and human coronary arteries, with only approximately 15% (mouse) and approximately 5% (human) of the enzyme being localized in mitochondria. Infection of ALDH2-deficient aortic smooth muscle cells or isolated aortas with adenovirus containing ALDH2 cDNA with or without the mitochondrial signal peptide sequence led to selective expression of the protein in mitochondria and cytosol, respectively. Cytosolic overexpression of ALDH2 restored GTN-induced relaxation and GTN denitration to wild-type levels, whereas overexpression in mitochondria (6-fold vs wild-type) had no effect on relaxation. Overexpression of ALDH2 in the cytosol of ALDH2-deficient aortic smooth muscle cells led to a significant increase in GTN denitration and cyclic GMP accumulation, whereas mitochondrial overexpression had no effect. The data indicate that vascular bioactivation of GTN is catalyzed by cytosolic ALDH2. Mitochondrial GTN metabolism may contribute to oxidative stress-related adverse effects of nitrate therapy and the development of nitrate tolerance.


The structure and stability of the monomorphic HLA-G are influenced by the nature of the bound peptide.
The highly polymorphic major histocompatibility complex class Ia (MHC-Ia) molecules present a broad array of peptides to the clonotypically diverse alphabeta T-cell receptors. In contrast, MHC-Ib molecules exhibit limited polymorphism and bind a more restricted peptide repertoire, in keeping with their major role in innate immunity. Nevertheless, some MHC-Ib molecules do play a role in adaptive immunity. While human leukocyte antigen E (HLA-E), the MHC-Ib molecule, binds a very restricted repertoire of peptides, the peptide binding preferences of HLA-G, the class Ib molecule, are less stringent, although the basis by which HLA-G can bind various peptides is unclear. To investigate how HLA-G can accommodate different peptides, we compared the structure of HLA-G bound to three naturally abundant self-peptides (RIIPRHLQL, KGPPAALTL and KLPQAFYIL) and their thermal stabilities. The conformation of HLA-G(KGPPAALTL) was very similar to that of the HLA-G(RIIPRHLQL) structure. However, the structure of HLA-G(KLPQAFYIL) not only differed in the conformation of the bound peptide but also caused a small shift in the alpha2 helix of HLA-G. Furthermore, the relative stability of HLA-G was observed to be dependent on the nature of the bound peptide. These peptide-dependent effects on the substructure of the monomorphic HLA-G are likely to impact on its recognition by receptors of both innate and adaptive immune systems.


Water lubricates hydrogen-bonded molecular machines.
The mechanical behaviour of molecular machines differs greatly from that of their macroscopic counterparts. This applies particularly when considering concepts such as friction and lubrication, which are key to optimizing the operation of macroscopic machinery. Here, using time-resolved vibrational spectroscopy and NMR-lineshape analysis, we show that for molecular machinery consisting of hydrogen-bonded components the relative motion of the components is accelerated strongly by adding small amounts of water. The translation of a macrocycle along a thread and the rotation of a molecular wheel around an axle both accelerate significantly on the addition of water, whereas other protic liquids have much weaker or opposite effects. We tentatively assign the superior accelerating effect of water to its ability to form a three-dimensional hydrogen-bond network between the moving parts of the molecular machine. These results may indicate a more general phenomenon that helps explain the function of water as the 'lubricant of life'.


Spontaneous production of interleukin 1 activity by chronic lymphocytic leukemic cells.
Chronic lymphocytic leukemic B cells (B-CLL) were found to produce an IL 1-like growth factor spontaneously in vitro for mouse thymocytes. This factor was comitogenic with concanavalin A (Con A) and nonmitogenic combinations of phorbol ester and calcium ionophore but not with phyto-hemagglutinin (PHA). Growth factor production was dose-related to the number of in vitro cultured cells and detectable at 6 hours using high cell concentrations. A small number of admixed normal T cells was not important for factor production. No growth of autologous B-CLL or allogeneic thymocytes was induced by the factor. A chromatographic high-performance liquid chromatography (HPLC) analysis and inhibition experiments with a polyclonal rabbit anti interleukin 1 (IL 1) antiserum indicated that the B-CLL-derived growth factor belonged to the IL 1 family. This was supported by the direct demonstration of IL 1 beta in supernatants from B-CLL by radioimmunoassay. Possible biologic implications for B-CLL-derived IL 1 are discussed in relation to tumor cell growth in different clinical stages.


Crystals of Rous sarcoma virus capsid protein show a helical arrangement of protein subunits.
Crystals of Rous sarcoma virus (RSV) capsid protein diffract X rays to 3.5 A resolution and belong to the monoclinic space group C2 with unit cell parameters a = 374.4 A, b = 128.1 A, c = 200.2 A, and beta = 121.8 degrees. One asymmetric unit of the crystal may contain between 28 and 35 molecules, based on reasonable crystal density assumptions. A self-rotation function and Patterson synthesis suggest that RSV capsid protein crystallizes as a helical array. The determinants of the viral particle morphology are not encoded in the capsid alone. The assembly of a helical array in the crystal reflects the absence of any conformational switching. However, it is expected that the subunit interactions seen in the crystal will be preferred and will relate to those found in the immature or mature virion.


Imaging the magnetic spin structure of exchange-coupled thin films.
We have investigated the magnetic spin structure of a soft-magnetic film that is exchange-coupled to a hard-magnetic layer to form an exchange-spring layer system. The depth dependence of the magnetization direction was determined by nuclear resonant scattering of synchrotron radiation from ultrathin 57Fe probe layers. In an external field a magnetic spiral structure forms that can be described within a one-dimensional micromagnetical model. The experimental method allows one to image vertical spin structures in stratified media with unprecedented accuracy.


Functionally independent subunits in the oligomeric structure of the GABA cotransporter rGAT1.
We have combined structural and functional approaches to investigate the role of oligomerization in the operation of the GABA transporter rGAT1. Xenopus laevis oocytes were induced to express, either separately or simultaneously, the wild-type form of rGAT1 and a mutated (Y140W) form, unable to translocate GABA and to generate transport currents, although its intramembrane charge movement properties are only slightly affected. These characteristics, together with the insensitivity of Y140W to the blocking action of SKF89976A, were used to study the possible functional interaction of the two forms in an heteromeric structure. The electrophysiological data from oocytes coexpressing wild-type and Y140W rGAT1 were consistent with a completely independent activity of the two forms. Oligomerization was also studied by fluorescence resonance energy transfer (FRET) in tsA201 cells expressing the transporters fused with cyan and yellow fluorescent proteins (ECFP and EYFP). All combinations tested (WT-ECFP/WTEYFP, Y140W-ECFP/Y140W-EYFP and WT-ECFP/ Y140W-EYFP) were able to give rise to FRET, confirming the formation of homo- as well as heterooligomers. We conclude that, although rGAT1 undergoes structural oligomerization, each monomer operates independently.


Pentoxifylline (Trental) decreases the replication of the human immunodeficiency virus type 1 in human peripheral blood mononuclear cells and in cultured T cells.
Pentoxifylline (Trental), used routinely for the treatment of intermittent claudication, has been shown previously to decrease the levels of tumor necrosis factors-alpha (TNF-alpha) RNA in cancer patients and to lead to a general improvement of well being. Increased TNF-alpha levels have been observed not only in cancer patients but also in cachectic patients with the acquired immunodeficiency syndrome (AIDS), and TNF-alpha is known to increase the expression of the human immunodeficiency virus type 1 (HIV-1) via activating its long terminal repeat (LTR). Moreover, TNF-alpha decreases the therapeutic efficacy of zidovudine (AZT). Here we show a significant decrease in HIV-1 replication by pentoxifylline in infected human peripheral blood mononuclear cells. The reduction was proportional to the downregulation of expression of a reporter gene, the bacterial gene for chloramphenicol acetyl transferase, linked to the HIV-1 LTR in human monocytoid cells. We conclude that patients with AIDS may benefit from pentoxifylline treatment because of its blockage of TNF-alpha-mediated HIV-1 upregulation, from increased efficacy of AZT, and also from improvement in TNF-alpha-induced cachexia.


Rapid detection of human cytomegalovirus in immunocompromised patients.
The Authors have evaluated, by DNA-probe hybridization, the presence of CMV in the cellular fractions of different body fluids from immunocompromised patients. This specific methodology appear to be rapid to perform, adaptable to different clinical samples and sensitive. Even in absence of a supporting evaluation of specific antibodies, direct detection of virus, by DNA-probe, seems to be the most efficient and rapid approach to confirm the diagnosis either of active phase or reactivating of CMV infection.


The inhibition of colorectal cancer growth by the natural product macrocarpal I.
Presently, few small molecule compounds are used as targeted therapy drugs in the treatment of colorectal cancer (CRC). It is important to identify new small molecule compounds, which can be used in the treatment of CRC. In this study, we selected four protein molecules as drug targets: PRL-3 (Phosphatase of regenerating liver 3), CLIC4 (Chloride intracellular channel 4), THBS2 (Thrombospondin 2), and BGN (Biglycan). These protein molecules were associated with the growth and metastasis of CRC cells. Small molecular compounds were screened on the basis of their target structures. Thus, five small molecule compounds were screened from each target structure, and three small molecule compounds (macrocarpal I, sildenafil, and neoandrographolide) were found to bind with two drug targets at the same time. Further experiments revealed that the inhibition rate of macrocarpal I was the highest in CRC cells. Therefore, we determined the effects of macrocarpal I on proliferation, apoptosis, cytoskeleton of CRC cells, and subcutaneous tumorigenesis in nude mice. Furthermore, RNA-seq analysis was performed to determine the molecular mechanism through which macrocarpal I inhibited the progression of CRC. We found that macrocarpal I could effectively inhibit proliferation, colony formation of CRC cells, and subcutaneous tumorigenesis in nude mice. Moreover, it also destroyed the cytoskeleton of CRC cells and promoted apoptosis. The effects on kinase activity, cytoskeleton, and DNA repair is the mechanism of macrocarpal I to inhibiting CRC growth. Macrocarpal I is a small molecule compound that can effectively inhibit the progression of CRC. Thus, macrocarpal I is a therapeutic compound that shows promising results in the treatment of advanced CRC.


Development of a new cellular immunological detection method for tuberculosis based on HupB protein induced IL-6 release test.
As a virulence factor, HupB plays important roles in the survival of MTB after infection and modulates the host immune response. In the current study, we aim to explore a new cellular immunological detection method for tuberculosis infection detection based on HupB protein. HupB was used to stimulate PBMCs extracted from pulmonary tuberculosis (PTB) patients, and secreted cytokines was examined. Then, we constructed a single center and a multi-center clinical trials to collect PBMCs from PTB patients, nPTB patients, or healthy volunteers to verify our findings. Cytokine's screening illustrated that IL-6 was the only cytokine released after HupB stimulation. Single-center and multi-center clinical trials showed that HupB stimulation significantly increased the level of IL-6 in the supernatant of PBMCs from PTB patients. Then we compared the specificity and sensitivity of HupB induced IL-6 release assay with ESAT-6 and CFP10 induced interferon γ release assay (IGRA), and found in smear positive PTB patients, the specificity and sensitivity of HupB induced IL-6 release assay was better than IGRA, and in smear negative PTB patients, the sensitivity was better. Combination of both assays provided an improved specificity and sensitivity for tuberculosis diagnosis. This study explored an immunological detection method for tuberculosis infection cells based on HupB protein-induced IL-6 release test, which can be used to enhance the diagnosis diagnostic accuracy of TB.


Single primer-mediated polymerase chain reaction: application in cloning of two different 5'-untranslated sequences of acidic fibroblast growth factor mRNA.
Polymerase chain reactions (PCR) are used to generate specific DNA sequences from minute amounts of DNA templates using a pair of oligonucleotide primers. To amplify regions of unknown sequence, methods such as inverted PCR, Alu PCR, and rapid amplification of cDNA ends (RACE) have been developed. These methods require several enzymatic manipulations of DNA which are either tedious or only suitable for certain special conditions. We have explored the possibility of PCR using a single primer. This method takes advantage of the fact that partial complementarity provides sufficient affinity for the oligonucleotide primer to anneal to a secondary, imperfect binding site. Thus, no modification of DNA template was required for the single primer-mediated PCR. We have used this method to generate two different aFGF cDNA clones containing different 5'-untranslated sequences.


Synergic influences of network topologies and associative interactions on the microstructures and bulk performances of hydrogels.
Revealing the relationship between network topologies and mechanical properties of hydrogels is fundamental yet challenging in the design of tough soft materials. Here, we report a series of hydrogels using N-isopropyl acrylamide (NIPAm) and acrylic acid (AAc) as the basic units to form a single network of the copolymer, a semi-interpenetrated network of two homopolymers, and a grafted network with homopolymer chains anchored on another homopolymer network, to investigate the influence of network architectures on the mechanical properties and thermal responses of the gels. We found that the properties of the gels are also significantly influenced by the formation of hydrogen bonds between poly(N-isopropyl acrylamide) (PNIPAm) and poly(acrylic acid) (PAAc) segments. The gels with the single network of poly(NIPAm-co-AAc) are mechanically weak due to the low efficiency for forming robust hydrogen bonds, while micro-segregated domains are formed in the hydrogels with a semi-interpenetrated network structure due to the formation of inter-chain hydrogen bonds that favors energy dissipation and toughening of the gels. On the other hand, dense hydrogen bonds form between the grafted PNIPAm chains and the PAAc network, resulting in nano-segregated domains and excellent mechanical properties of the gels. The hydrogels with the grafted network structure exhibit a more repeatable response to temperature than those with the semi-interpenetrated network structure due to the relatively stable hydrogen-bond network. The comparison of the mechanical properties and thermal stability of the hydrogels with the same composition but different topological networks should be informative for engineering hydrogel properties or functions by tailoring the network structures.


Simplified Approach to Work Function Modulation in Polyelectrolyte Multilayers.
The layer-by-layer (LbL) method is based on sequential deposition of polycations and polyanions. Many of the properties of polyelectrolyte thin films deposited via this method depend on the nature of the topmost layer. Thus, these properties show odd-even oscillations during multilayer growth as the topmost layer alternates from polycations to polyanions. The work function of a (semi)conductive substrate modified with an LbL polyelectrolyte multilayer also displays an oscillatory behavior independent of film thickness. The topmost layer modulates the work function of a substrate buried well below the film. In agreement with previous observations, in this work, we show that the work function of a gold substrate changes periodically with the number of adsorbed layers, as different combinations of polycations and polyanions are deposited using the LbL method. For the first time, we rationalize this behavior in terms of formation of a dipole layer between the excess charge at the topmost layer and the charge of the metal substrate, and we put forward a semiquantitative model based on a continuum description of the electrostatics of the system that reproduces the experimental observations.


[Treatment of distal humerus fracture with unexposed ulnar nerve medial elbow incision and anatomical locking compression plate].
To investigate the feasibility and effectiveness of unexposed ulnar nerve medial elbow incision, open reduction and internal fixation of anatomical locking compression plate (LCP) for distal humerus fractures. Fourteen patients with distal humerus fracture were treated between January 2014 and June 2017. There were 5 males and 9 females, aged 18-85 years (mean, 65.5 years). The causes of injury included falling from height in 12 cases and traffic accident in 2 cases, all were closed fractures. Fractures were classified according to the AO/Association for the Study of Internal Fixation (AO/ASIF): 3 cases of type A2, 2 cases of type A3, 4 cases of type B2, 2 cases of type C1, 2 cases of type C2, and 1 case of type C3; without ulnar nerve damage. The time from injury to operation was 4-15 days, with an average of 7 days. The type B2 fractures were treated with unexposed ulnar nerve elbow medial incision and anatomic LCP internal fixation, the rest patients were all treated with unexposed ulnar nerve medial plus conventional lateral approach and bilateral LCP internal fixation. The operation time was 50-140 minutes (mean, 80 minutes), and the intraoperative blood loss was 20-200 mL (mean, 70 mL). There was no blood vessels or nerve damage during operation. All incisions healed by first intension, and no incision infection occurred. All the 14 cases were followed up 9-24 months (mean, 13 months). X-ray films showed that all fractures healed within 4 months without complications such as nonunion and osteomyelitis. No ulnar nerve injury, cubitus varus deformity, and ossifying myositis occurred during follow-up. At last follow-up, the elbow function was assessed by Mayo Elbow Performance score (MEPS), the results were excellent in 8 cases, good in 4 cases, fair in 1 case, and poor in 1 case (type C3 fracture), with the excellent and good rate of 85.7%. The unexposed ulnar nerve medial elbow incision can be used effectively to reduct the fracture, and it is not prone to ulnar nerve injury. Combined with the lateral approach to treat the distal humerus fracture, which has the advantages of short operation time, few trauma, little bleeding, and reliable effectiveness.


Students' responses to the introduction of a digital laboratory guide in medical neuroscience.
This study investigated student acceptance of a digital laboratory guide in a medical neuroscience course. The guide was created in Macromedia Authorware 5.2 by transforming a text-based lab manual into a comprehensive multimedia program. Student attitudinal survey data indicated that the guide was used primarily as a study tool at home. Some 53% of students (primarily males) used the guide for independent study exclusively, while the remainder used it for both independent and group study; 87% of the class used the guide as a routine study tool rather than as a final review for examinations. Most students perceived that the guide increased their study efficiency and lab performance while decreasing their out-of-class study time in the lab. Female students were significantly more enthusiastic about the guide's impact on their study efficiency, lab performance and laboratory study time. Gender differences in student satisfaction with and use of instructional multimedia are discussed.


Count rate performance of brain-dedicated PET scanners: a Monte Carlo simulation study.
In recent years, there has been a renewed interest in brain-dedicated PET imaging systems, particularly in the context of combined PET/MR imaging. We are currently designing a brain-dedicated PET insert suitable for an ultra-high field brain-dedicated MR scanner, the Siemens Magnetom 7T MR scanner. In this paper, an investigation on the count rate performance of several possible detectors through a series of Monte Carlo simulations is reported. Brain-dedicated PET scanners with a lutetium oxyorthosilicate scintillator and a detector area of 0.04 (1 crystal per detector) to 101.37 (2500 crystals per detector) cm2, detector thickness of 10 to 20 mm and a fixed crystal pitch of ~2 mm were simulated. The count rate performance of each scanner was evaluated as a function of detector deadtime type and constant, coincidence timing window and lower level discriminator. Also, the effects of activity outside the field-of-view (FOV) on the count rate performance of each scanner were studied. For each detector geometry and performance metric, the scanner singles rate, scanner sensitivity and noise equivalent count rate as a function of activity in the FOV were measured. It was seen that scanners with detectors comprised a few crystal elements showed reduced scanner sensitivity due to a high number of inter-detector scattering. The count rate performance of scanners with large detectors, on the other hand, was mainly determined by the deadtime properties of the detectors. A model for the count rate performance of the scanner with each studied detector is presented in this work.


[Long-term results of one stage tracheoesophageal end-side anastomosis voice restoration after total laryngectomy].
The purpose of this study is to discuss long-term results of one different kind of voice restoration after total laryngectomy with 10 years experience. One stage tracheoesophageal end-side anastomosis after total laryngectomy was performed in 36 patients from 1991-1996. The key technique of this operation is to make a tongue-ship flap(1.5 cm x 1.5 cm-1.8 cm x 1.5 cm) with tracheal membranous part and dissect it from the esophagus to 1 cm below the upper incisal margin of trachea. Then a 0.8 cm middle vertical incision was made on the lowest anterior wall of the separated esophagus. Folding the tongue-ship flap posteriorly to approximate the esophagus incision and make an end-side anastomosis. Close the upper incisal margin of trachea with left to right direction suture. The follow up time is 5 - 10 year. One lost follow 2.5 years later and was judged as dead. The short-term and long-term success rate were 69% (25/36) and 83% (30/36) respectively. The 3-, 5- and 10 year survival rate are 72%, 69%, 69%, respectively. This technique does not change the pattern of total laryngectomy and various neck dissections do not affect voice rehabilitation. Advantages of the operation: easy to operate and learn; the special form of tracheoesophageal shunt (high in the tracheal side and low in esophageal side) prevents the aspiration; high successful rate, excellent speech, fluent language, good acceptability and lower aspiration.


Bioenergetics of Monoterpenoid Essential Oil Biosynthesis in Nonphotosynthetic Glandular Trichomes.
The commercially important essential oils of peppermint (Mentha × piperita) and its relatives in the mint family (Lamiaceae) are accumulated in specialized anatomical structures called glandular trichomes (GTs). A genome-scale stoichiometric model of secretory phase metabolism in peppermint GTs was constructed based on current biochemical and physiological knowledge. Fluxes through the network were predicted based on metabolomic and transcriptomic data. Using simulated reaction deletions, this model predicted that two processes, the regeneration of ATP and ferredoxin (in its reduced form), exert substantial control over flux toward monoterpenes. Follow-up biochemical assays with isolated GTs indicated that oxidative phosphorylation and ethanolic fermentation were active and that cooperation to provide ATP depended on the concentration of the carbon source. We also report that GTs with high flux toward monoterpenes express, at very high levels, genes coding for a unique pair of ferredoxin and ferredoxin-NADP+ reductase isoforms. This study provides, to our knowledge, the first evidence of how bioenergetic processes determine flux through monoterpene biosynthesis in GTs.


Cystatin C Concentration During the First Three Postnatal Days in Healthy Term Newborns.
Cystatin C (Cys-C) concentration has not been examined sufficiently among healthy newborn population, particularly in terms of reference values. This study aimed to establish gender-, postnatal age- and birth weight-specific Cys-C concentration for healthy term newborns. Its objective was also to examine if there were any differences between our measured concentration and the reference interval established by the CALIPER study. Serum samples from a total of 90 healthy term newborns were used to determine Cys-C concentration. Cys-C was measured within first three days of birth using particle-enhanced turbidimetric immunoassay (PETIA) on the Architect plus ci8200 analyzer. Median concentration of the Cys-C was 2.05 mg/L. There were no statistically significant differences in Cys-C concentration regarding gender (p=0.779), birth weight (p=0.505), postnatal age (p=0.512) or Apgar score (p=0.799). The value of the 2.5th and 97.5th percentile for Cys-C concentrations for girls was 0.93-3.15 mg/L and for boys it was 1.5-3.36 mg/L. Cys-C concentration in healthy term newborns does not depend on gender, birth weight, postnatal age, or Apgar score. Our measured concentration range of CyS-C in healthy newborns turned out to be slightly wider than the interval determined in the CALIPER study.


Development of a quantitative in vitro transformation assay for kidney epithelial cells.
A quantitative in vitro transformation assay has been developed for the first time using primary rat kidney epithelial (RKE) cells. RKE cells were grown in a 50:50 mixture of 3T3 conditioned medium and DF8 medium composed of Ham's F-12/DMEM supplemented with ferrous sulfate, vasopressin, transferrin, sodium selenite and 10% fetal bovine serum. Colony forming efficiency of cells plated in this medium was high, ranging from 2.4 to 16%. Normal RKE cells treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) became transformed to a preneoplastic state of enhanced in vitro growth potential and formed large colonies of morphologically altered cells, whereas RKE cells treated with vehicle alone ceased proliferating and/or sloughed off the dish within 4-6 weeks. Relative survival and percent transformation frequency (Tf) of RKE cells exposed to MNNG were inversely proportional and both were dose dependent. MNNG concentration of 0.5, 1.0 and 2.0 micrograms/ml resulted in transformation frequencies of 0.13, 0.37 and 1.1% respectively (n = 4). Morphologically transformed colonies gave rise to cell lines with indefinite growth capacity and neoplastic potential. One of six transformed RKE cell (TRKE) lines injected into nude mice produced adenocarcinomas. This assay represents the first in vitro model for studying mechanisms of chemical transformation of normal kidney epithelial cells and may also be useful as a screen for identifying potential renal carcinogens.


Treating alcoholics and their spouses: a family-systems centred approach.
The need and rationale for an expanded focus in the treatment of alcoholism are outlined in this paper. The inadequacy of a treatment regime which treats alcoholics in isolation from their families is demonstrated. The author describes the Couples Treatment Program in Regina, which illustrates a new and innovative approach to dealing with underlying issues related to drinking behaviour. This approach recognizes that marital and family issues and perpetuation of drinking are highly interrelated. Recovery involves addressing not only the drinking problem, but also relationship issues that reinforce drinking. The role of the spouse of the alcoholic in treatment and the need for treatment of the entire nuclear family are also mentioned.


[40 years to Flegel's disease (hyperkeratosis lenticularis perstans)].
A case report of hyperkeratosis lenticularis perstans (Flegel) is presented. The disease is characterized clinically, histologically and with electron microscope. The evolution of the disease since Flegel described it 40 years ago is considered, with emphasis on etiology, pathogenesis and unanswered questions.


Chimeric tRNAs as tools to induce proteome damage and identify components of stress responses.
Misfolded proteins are caused by genomic mutations, aberrant splicing events, translation errors or environmental factors. The accumulation of misfolded proteins is a phenomenon connected to several human disorders, and is managed by stress responses specific to the cellular compartments being affected. In wild-type cells these mechanisms of stress response can be experimentally induced by expressing recombinant misfolded proteins or by incubating cells with large concentrations of amino acid analogues. Here, we report a novel approach for the induction of stress responses to protein aggregation. Our method is based on engineered transfer RNAs that can be expressed in cells or tissues, where they actively integrate in the translation machinery causing general proteome substitutions. This strategy allows for the introduction of mutations of increasing severity randomly in the proteome, without exposing cells to unnatural compounds. Here, we show that this approach can be used for the differential activation of the stress response in the Endoplasmic Reticulum (ER). As an example of the applications of this method, we have applied it to the identification of human microRNAs activated or repressed during unfolded protein stress.


Surgical treatment of a left ventricular neurofibroma.
Primary cardiac neurofibroma is a rare occurrence. We describe a case of left ventricular neurofibroma in a 56-year-old woman with Von Recklinghausen disease. Resection of the tumor with concomitant mitral valve replacement yielded a satisfactory clinical result, and histological examination of the resected tissue confirmed benign neurofibroma. The anatomic distribution of the vagus nerve plexus, which penetrates the epicardium and myocardium and courses over the left ventricular subendocardial surface, provides a tissue source from which this neurogenic tumor may arise.


Cell-mediated immune response to copolymer I in multiple sclerosis measured by the macrophage procoagulant activity assay.
The macrophage/monocyte procoagulant activity (MPCA) assay, a sensitive and specific in vitro test for cell-mediated immunity, has been used to ascertain the reactivity of MS peripheral blood mononuclear cells (PBM) to copolymer I (Copl), a synthetic peptide analogue of myelin basic protein (MBP) currently being tested as a possible therapeutic agent in multiple sclerosis (MS). Because the suppressive effect of Copl is believed to lie in its possible cross-reactivity with MBP, the reactivity of PBM of MS patients to MBP was also tested. MS patients either at the stable phase of the relapsing/remitting form or with chronic progressive disease were investigated and compared with patients with other diseases and with healthy subjects. The reactivity to Copl was significantly increased in patients with chronic progressive disease but not in stable MS patients or in control subjects. No difference in reactivity to MBP between MS patients and healthy subjects was found regardless of disease status. However, in the control group comprising patients with other diseases, MBP reactivity was significantly elevated. In chronic progressive MS patients, a relationship was found between the response to Copl and that to MBP, supporting the possibility of an immunological cross-reactivity between these two antigens. There was no significant difference in reactivity to the non-specific stimulant, lipopolysaccharide, between the MS and control groups.


Description of the pupa of Aedes pseudotaeniatus (Giles) (Diptera: Culicidae).
The pupal stage of Aedes pseudotaeniatus (Giles) is described for the first time. Collections of this species were made in the Himalayan Kingdom of Nepal. Group E of subgenus Finlaya is discussed.


Polarized wavelength-dependent measurements of turbid media.
Wavelength-dependent, polarized, elastic-scatter spectra of tissue phantoms and in vitro tissue are presented. These measurements are shown to be sensitive to very small changes in composition of the scattering medium. A simple physical explanation of the wavelength-dependent polarization phenomena observed for media consisting only of spherical particles is given and the relevance of wavelength-dependent, polarized, elastic-scatter spectra to in vivo applications are discussed.


HMGA2 Immunoexpression is frequent in salivary gland pleomorphic adenoma: immunohistochemical and molecular analyses of PLAG1 and HMGA2 in 25 cases.
The main oncologic events in pleomorphic adenoma (PA) are the translocations of Pleomorphic adenoma gene 1 (PLAG1) on chromosome 8q12 and High-mobility group AT-hook 2 (HMGA2) on chromosome 12q14.3 with various fusion partners. These translocations result in the transcriptional up-regulation of PLAG1 and HMGA2 proteins. We carried out a preliminary evaluation of PLAG1 translocation by fluorescence in-situ hybridization (FISH), immunohistochemistry (IHC) and HMGA2 IHC on twenty-five archived formalin-fixed paraffin-embedded tissues of PAs and its clinicopathologic features. Only eight cases were successfully hybridized and 50% of the interpretable cases were considered positive for PLAG1 translocation. PLAG1 IHC was only positive in 2 (8%) of the 25 cases stained, including one of the positive PLAG1 translocation cases. HMGA2 IHC was positive in 12 (48%) of the 25 cases stained including 2 (50%) of the 4 cases identified with PLAG1 translocation by FISH, 3 (75%) of the 4 cases negative for PLAG1 translocation by FISH and 7 (41%) of the 17 cases with failed hybridization. Overall, 15 (60%) of the 25 PA cases demonstrated PLAG1 and/or HMGA2 alterations confirmed either by FISH or IHC. In conclusion, PLAG1 and HMGA2 alterations were confirmed either by FISH or IHC in this cohort and HMGA2 alteration is a common event in PAs of salivary gland.


[Reception and signal transduction implied in the differentiation of stem cells from human bone marrow until adipocytes].
Glucagon-like peptide-1 (GLP-1) functions as an incretin hormone with antidiabetogenic properties. However the role of GLP-1 in human bone marrow-derived mesenchymal stem cells (hMSCs), if any, remains unknown. The effects of GLP-lin hMSCs were tested with regard to cell proliferation, cytoprotection, and cell differentiation into adipocytes. The signalling pathways involved in these processes were also analyzed. Cells were characterized by biochemical and morphological approaches before and after being induce to differentiate into adipocytes. PCNA protein levels were used as an protein index, whereas cell apoptosis was studied by deprivation of fetal bovine serum. Isolated hMSCs expressed stem cell markers as well as mRNA and GLP-1 receptor protein. GLP-1 increased the proliferarion of hMSCs, which decreased when they were induced to differentiate into adipocytes.This process produced biochemical and morphological changes in cells expressing PPARgamma, C/EBPbeta, AP2, and LPL in a time-dependent pattern. Notably, GLP-1 significantly reduced the expression of PPARgamma, C/EBPbeta, and LPL. These effects were exerted at least through the MEK and PKC signaling pathways. In addition, GLP-1 significantly reduced cell apoptosis. Our data indicate that, in hMSCs, GLP-1 promotes cellular proliferation and cytoprotection and prevents cell differentiation into adipocytes. These latter findings underscore the potential therapeutic role of GLP-1 in preventing the adipocyte hyperplasia associated with obesity and, additionally, could bolster the maintenance of hMSC stores by promoting the proliferation and cytoprotection of undifferentiated hMSC. These data suggest that the GLP-1 may have some function during embryonic life. Thus, both the peptide and its receptor were identified early in the mouse development, as well as GLP-1 modified significantly gene expression in mouse undifferentiated pluripotent cells. These findings also suggest that the pathophysiological implications of GLP-1 imbalance in adulthood may have a counterpart during development.


Influence of Helicteres isora administration for diabetes mellitus: Its effect on erythrocyte membrane and antioxidant status.
In this study the effect of Helicteres isora L. on erythrocyte membrane bound enzymes and antioxidants activity in plasma and erythrocytes of streptozotocin (STZ) induced diabetic model was investigated. The aqueous bark extract of H. isora was administered orally for 30 days to control and STZ induced diabetic rats. The effect of bark extract on glucose, insulin, haemoglobin, glycosylated haemoglobin, TBARS, hydroperoxide, superoxide dismutase (SOD), catalase (CAT), glutathione peroxide (GPx), glutathione-S-transferase (GST), vitamins C and E, reduced glutathione (GSH) and membrane bound enzymes were studied. The levels of glucose, glycosylated haemoglobin, TBARS, hydroperoxide, and vitamin E were increased significantly whereas the level of insulin, haemoglobin, as well as antioxidants, membrane bound total ATPase, Na(+)/K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase were decreased significantly in STZ diabetic rats. Administration of bark extract to diabetic rats showed a decrease in the levels of glucose, glycosylated haemoglobin, lipid peroxidation markers and vitamin E. In addition the levels of insulin, haemoglobin, enzymatic antioxidants, vitamin C, and GSH and the activities of membrane bound enzymes also were increased in H. isora treated diabetic rats. The present study indicates that the H. isora possesses a significant favourable effect on erythrocyte membrane bound enzymes and antioxidants defense system in addition to its antidiabetic effect.


alpha-Melanocyte-stimulating hormone-induced eruptive nevi.
Synthetic peptides that target proopiomelanocortin receptors are being investigated as a novel and safe way to tan. It has been postulated that synthetic alpha-melanocyte-stimulating hormone (alpha-MSH) peptides may have protective effects against the development of melanoma because of their melanogenic activity. Their ultimate biological effect, however, especially in patients with dysplastic nevi or previous melanoma, has yet to be determined. A 40-year-old white man with a history of melanoma and multiple dysplastic nevi self-administered synthetic alpha-MSH. He developed crops of new pigmented nevi, many of which had atypical clinical and histopathologic features. The preexisting nevi became darker and acquired growth features. After alpha-MSH use was discontinued, the nevi progressively lightened and lost their growth features. Synthetic alpha-MSH peptides can drive proliferation of neoplastic melanocytic cells in predisposed patients. This could present an increased risk for melanoma development.


Clinical carotid atherosclerosis.
Atherogenesis is an inflammatory process that begins in childhood. Early detection of atherosclerosis might allow for selection of subjects at risk for future cerebrovascular events at a time when dietary and lifestyle modification may have its greatest impact, and medical intervention may be useful for those who are refractory to such treatment or who are at greater risk for an event. Early detection relies on a knowledge of the distribution of atheroma formation, which can be predicted by understanding the hemodynamic patterns of blood flow. Early formation is accompanied by a vascular remodeling that normalizes the area of the vessel lumen, making early detection impossible by angiography. Elevated serologic markers of inflammation may be used as evidence of formation of atherosclerosis, but inflammatory markers lack sensitivity and specificity. Preliminary evidence supports the ability of MRI to detect early atheroma formation, possibly even before substantial wall thickening occurs. Once atheroma has formed with measurable stenosis, the goal of imaging studies is to predict stroke risk and determine the need for surgical intervention. Subjects with symptomatic high-grade carotid stenosis have been shown to benefit from surgery with a reduced stroke risk when compared with medical management alone, although controversy exists regarding the management of moderate symptomatic carotid stenosis or asymptomatic carotid disease. In these individuals, understanding atheroma morphology may prove to be of greater utility for assessing stroke risk and determining the appropriate management. Morphologic characterization may also be helpful in monitoring the effect of medical intervention. MRI has proven capable of characterizing the morphologic composition of carotid atheroma, although the clinical implications continue to be investigated.


Transcription Factor VviMYB86 Oppositely Regulates Proanthocyanidin and Anthocyanin Biosynthesis in Grape Berries.
Proanthocyanidins (PAs) and anthocyanins are two vital groups of flavonoid compounds for grape berries and red wines. Several transcription factors (TFs) have been identified to be involved in regulating PA and anthocyanin biosynthesis in grape berries. However, research on TFs with different regulatory mechanisms for these two biosynthesis branches in grapes remains limited. In this study, we identified an R2R3-MYB TF, VviMYB86, whose spatiotemporal gene expression pattern in grape berries coincided well with PA accumulation but contrasted with anthocyanin synthesis. Both in vivo and in vitro experiments verified that VviMYB86 positively regulated PA biosynthesis, primarily by upregulating the expression of the two leucoanthocyanidin reductase (LAR) genes in the Arabidopsis protoplast system, as well as in VviMYB86-overexpressing grape callus cultured under 24 h of darkness. Moreover, VviMYB86 was observed to repress the anthocyanin biosynthesis branch in grapes by downregulating the transcript levels of VviANS and VviUFGT. Overall, VviMYB86 is indicated to have a broad effect on flavonoid synthesis in grape berries. The results of this study will help elucidate the regulatory mechanism governing the expression of the two LAR genes in grape berries and provide new insights into the regulation of PA and anthocyanin biosynthesis in grape berries.


The diagnosis less traveled: NPs' role in recognizing adult ADHD.
To stimulate critical thought about sociocultural implications of unrecognized and undiagnosed attention deficit hyperactivity disorder (ADHD), and how these factors interface with healthcare delivery models and care that nurse practitioners (NPs) provide. Health science databases--Psych Info, Proquest, Sage, PubMed, and authors' professional experiences. NPs, often the main healthcare provider for underserved populations in community practice settings, have little training in assessing adult ADHD. ADHD, often unrecognized and undiagnosed among adults, contributes to global impairments adversely affecting individuals' social, behavioral, academic, and cognitive functioning. Increased insight and awareness about adult ADHD is warranted to facilitate appropriate diagnosis. ADHD is found in all sectors of our society; however, assessment and diagnosis among those whose socioeconomic status limits access to resources is a problem. Working in integrated care clinical settings facilitates recognition of patient problems and colocates resources required to manage the ADHD patient effectively. While this practice model may not be the norm, it is critical for NPs to have: (a) heightened awareness of the presentation of adult ADHD; (b) skills and/or resources to facilitate proper diagnosis of adult ADHD, and (c) models of practice that support optimal NP care delivery.


Budd-Chiari syndrome: An unusual complication of an internal jugular tunneled dialysis catheter.
Budd-Chiari syndrome due to the tip of an internal jugular tunneled dialysis catheter malposition in inferior vena cava or hepatic vein is a rare complication. We aimed to present our experience and compare it with the previous reports to highlight the clinical features and the optimal management. A 57-year-old female with history of ANCAp vasculitis, treated by hemodialysis in the last 2 years on a right internal jugular vein tunneled catheter was admitted for pain in the right upper quadrant. A subacute Budd-Chiari syndrome due to catheter malposition was diagnosed. The catheter was removed, and a new tunneled hemodialysis line was inserted in the right internal jugular vein with the tip at the junction of right atrium with superior vena cava. Anticoagulation with apixaban 2.5 mg twice daily was started after catheter replacement and the patient was discharged. At 1 month follow-up the patient had no symptoms, and the ultrasound revealed the absence of the thrombus in the inferior vena cava. Imagining monitoring for malposition after insertion or in a clinical context suggestive for Budd-Chiari syndrome is essential for early diagnosis and treatment. In our case, anticoagulation with apixaban and prompt catheter replacement resulted in Budd-Chiari syndrome resolution.


Thiopental inhibits global protein synthesis by repression of eukaryotic elongation factor 2 and protects from hypoxic neuronal cell death.
Ischemic and traumatic brain injury is associated with increased risk for death and disability. The inhibition of penumbral tissue damage has been recognized as a target for therapeutic intervention, because cellular injury evolves progressively upon ATP-depletion and loss of ion homeostasis. In patients, thiopental is used to treat refractory intracranial hypertension by reducing intracranial pressure and cerebral metabolic demands; however, therapeutic benefits of thiopental-treatment are controversially discussed. In the present study we identified fundamental neuroprotective molecular mechanisms mediated by thiopental. Here we show that thiopental inhibits global protein synthesis, which preserves the intracellular energy metabolite content in oxygen-deprived human neuronal SK-N-SH cells or primary mouse cortical neurons and thus ameliorates hypoxic cell damage. Sensitivity to hypoxic damage was restored by pharmacologic repression of eukaryotic elongation factor 2 kinase. Translational inhibition was mediated by calcium influx, activation of the AMP-activated protein kinase, and inhibitory phosphorylation of eukaryotic elongation factor 2. Our results explain the reduction of cerebral metabolic demands during thiopental treatment. Cycloheximide also protected neurons from hypoxic cell death, indicating that translational inhibitors may generally reduce secondary brain injury. In conclusion our study demonstrates that therapeutic inhibition of global protein synthesis protects neurons from hypoxic damage by preserving energy balance in oxygen-deprived cells. Molecular evidence for thiopental-mediated neuroprotection favours a positive clinical evaluation of barbiturate treatment. The chemical structure of thiopental could represent a pharmacologically relevant scaffold for the development of new organ-protective compounds to ameliorate tissue damage when oxygen availability is limited.


Elevation of cerebrospinal fluid soluble CD27 levels in patients with meningeal localization of lymphoid malignancies.
Diagnosis of meningeal localization of lymphoid malignancies by means of cytologic examination of the cerebrospinal fluid (CSF) can be difficult. Thus far no reliable CSF tumor markers have been identified. CD27 is a transmembrane disulfide-linked 55-kD homodimer present on most peripheral blood T cells and on a subset of B cells. CD27 is also expressed on human malignant B cells and high levels of soluble CD27 can be present in the serum of patients with B-cell malignancies. The aim of this study is to determine prospectively the diagnostic value of CSF sCD27 as a tumor marker in patients with meningeal localization of lymphoid malignancies. CSF sCD27 levels were determined by sandwich enzyme-linked immunosorbent assay. The optimal cut-off value using receiver operator characteristics curves was found to be 10 U/mL. sCD27 levels were normal in all 50 control patients (lumbar disc protrusion) and in 39 of 40 samples obtained from patients with either solid tumors or acute myeloid leukemia. Of 104 CSF samples from 70 children with acute lymphoblastic leukemia (ALL) or non-Hodgkin's lymphoma (NHL) undergoing routine central nervous system (CNS) staging, sCD27 was false positive and false negative in only one sample each. In 70 samples from 45 patients suspected of meningeal localization of ALL or NHL, the sCD27 test had an excellent sensitivity (100%) and specificity (82%). In 7 patients with positive CSF studied longitudinally, sCD27 levels correlated very well with remission and relapse. sCD27 levels were not nonspecifically increased by the administration of cytostatic drugs. Finally, sCD27 was also elevated in the 4 patients studied with primary central nervous system lymphoma (PCNSL). CSF sCD27 is a promising tumor marker in patients with either meningeal localization of lymphoid malignancies or PCNSL, and can be useful in the differential diagnosis of CNS involvement by either lymphoid malignancies or solid tumors.


[Studies on extraction of Ligusticum chuanxiong in Hongye Xintong soft capsule by super critical fluid extraction].
To study the rationality of extraction prosess for the Ligusticum chuanxiong in Hongye Xintong Soft Capsule by super critical fluid extraction(SFE). Conditions for the extraction were optimized by orthogonal experimental design as guided by the extraction rate and content of ligustilid in the extract; Combined with the experiment of rats ligated the left coronary artery, the two compounds containing different extracts (steam distillation and SFE) were compared to determine the extraction prosess. The best extraction conditions were established as following: pressure 30 MPa, temperature 50 degrees C, extracting time 4 h, separate pressure 10 MPa. separate temperature 40 degrees C. Compared with steam distillation, the compound containing SFE extract showed stronger protective effects on rats ligated the left coronary artery. Super critical fluid can extract active ingredients in Ligusticum chuanxiong effectively.


Modeling of the Charge-Transfer-Induced Spin Transition in the Tetranuclear Cyanide-Bridged [Co2Fe2] Complex.
The course of the charge-transfer-induced spin transition demonstrated by the cyanide-bridged tetranuclear [Co2Fe2(bpy*)4(CN)6(tp*)2](PF6)2·2CP·8BN complex has been followed by DFT calculations of the single-point energies for different total spin values of the complex in a wide temperature range. With the aid of these calculations, the picture of spin conversion, that the compound undergoes, has been restored. It has been demonstrated that at 100 K the two crystallographically unique tetranuclear Fe2Co2 subunits A and B present in the structure contain diamagnetic low-spin FeII and low-spin CoIII ions. From the three subunits A, A', and B detected crystallographically in the compound at 200 K, only the A ones contain paramagnetic low-spin FeIII and high-spin CoII ions, while at 260 K, a half of all clusters contained in the crystal are in this state. From the DFT calculations, it also follows that at 320 K in the crystal only paramagnetic units are present. The results obtained are in accordance with the experimental data on the magnetic susceptibility. A possibility to predict new materials exhibiting spin transitions on the basis of DFT calculations of single-point energies as functions of temperature is discussed.


Reactive Oxygen Species and Nitric Oxide Control Early Steps of the Legume - Rhizobium Symbiotic Interaction.
The symbiotic interaction between legumes and nitrogen-fixing rhizobium bacteria leads to the formation of a new organ, the nodule. Early steps of the interaction are characterized by the production of bacterial Nod factors, the reorientation of root-hair tip growth, the formation of an infection thread (IT) in the root hair, and the induction of cell division in inner cortical cells of the root, leading to a nodule primordium formation. Reactive oxygen species (ROS) and nitric oxide (NO) have been detected in early steps of the interaction. ROS/NO are determinant signals to arbitrate the specificity of this mutualistic association and modifications in their content impair the development of the symbiotic association. The decrease of ROS level prevents root hair curling and ITs formation, and that of NO conducts to delayed nodule formation. In root hairs, NADPH oxidases were shown to produce ROS which could be involved in the hair tip growth process. The use of enzyme inhibitors suggests that nitrate reductase and NO synthase-like enzymes are the main route for NO production during the early steps of the interaction. Transcriptomic analyses point to the involvement of ROS and NO in the success of the infection process, the induction of early nodulin gene expression, and the repression of plant defense, thereby favoring the establishment of the symbiosis. The occurrence of an interplay between ROS and NO was further supported by the finding of both S-sulfenylated and S-nitrosylated proteins during early symbiotic interaction, linking ROS/NO production to a redox-based regulation of the symbiotic process.


Verminous myelitis in a pit bull puppy.
A 10-week-old, male pit bull dog presented to the referring veterinarian with hind limb paresis and epaxial muscle atrophy. No spinal lesions were identified at gross necropsy; however, histologically there was marked granulomatous myelitis in the spinal cord between T13 and L2 with occasional, intralesional nematode larvae. Based on morphologic characteristics, the nematode larvae were identified as Strongyloides spp., possibly Strongyloides stercoralis.


A case of acute psychosis in a patient following exposure to a single high dose of styrene.
We report a case of acute psychotic symptoms following exposure to a single high dose of styrene monomer. The 24-year-old male patient showed psychotic and cognitive symptoms immediately after exposure. His psychotic symptoms included auditory hallucinations and delusions of reference. Brain magnetic resonance imaging, electroencephalography, and laboratory examinations were performed to evaluate any other causes. The clinical, neuroimaging, and laboratory review in this case suggested that the suddenly developed psychotic symptoms that led to chronic deterioration were caused by the single exposure to styrene monomer. This is the first recent report in which acute psychotic symptoms developed from a single high dose of styrene suffocation compared with previous findings showing symptoms because of long-term low-dose exposure.


Sucralfate in gastritis.
Gastritis has a wide spectre of definition modalities. Most previous studies have compared symptomatology with histologic gastritis with negative results. We believe that this may be due to inadequate definition criteria and emphasize this point by comparing gastroesophageal reflux with duodenogastric reflux. A prospective randomized trial has been conducted for half a year comparing Sucralfate with a placebo in patients with symptomatological and macroscopical gastritis. Although approximately one hundred patients met the endoscopic criteria, the vast majority could not be included due to well-defined interfering diseases, and thus the material is still too sparse to give any indication of the influence of Sucralfate on endoscopic gastritis, although the preliminary overall results seem promising.


[THE HEALTH OF THE POPULATION, ACCORDING TO PRENOSOLOGICAL DIAGNOSTICS].
Traditional methods of medical examination used during medical visits give little information about the adaptive capabilities of the body, which largely determine the body's ability to adapt to changing environmental conditions. Therefore, diagnosis of adaptive capacity of the body, which can be attributed to the form of prenosological diagnosis of the identified changes, is necessary for an objective assessment of health and improving the effectiveness of health measures. In our work we used the methodology of G. L. Apanasenko, E. A. Pirogova, Berseneva A. B. and Y. P. Suicine. The study involved 1010 people of the population of Astana and Northern regions of Kazakhstan. In all age categories, the higher level of physical condition was among women (64.15±1.95%), compared with men (20.83±2.04%). There were no significant differences in the values of adaptive capacity among men and women, but the number of women with high adaptive capacity (62.7±1.96%) was higher than that of men (55.99±2.5%). The index of physical condition and adaptive capacity were higher among the adolescent population (84.4±4.05%) and had a natural tendency to decrease with age. In 74.4±0.31% of cases it was caused by deterioration of functional parameters of cardiovascular and respiratory systems. Comparative analysis of the adaptive capacity of the population in the regions showed their unidirectional change with the index of physical condition. The lowest values of adaptive potential, indicating a high degree of adaptation, were residents of Astana (54.56±3.52%). The worst performance was observed among the villagers Zarja and Ubaganskoe. In our opinion, such differences may be due to differences in income levels, education, as well as the availability of sports and recreation facilities.


Purification of myo-Inositol 1-Phosphate Synthase from Rice Cell Culture by Affinity Chromatography.
myo-Inositol 1-phosphate synthase (EC 126.96.36.199) is the enzyme which catalyzes the synthesis of the precursor for the myo-inositol oxidation pathway. Rice callus grown in suspension culture provides a good source of plant enzyme. Use has been made of a noncompetitive inhibitor to prepare an affinity column for this enzyme. With this column, the enzyme from rice callus has been purified 1500-fold in a single step, about 9000-fold over-all, to a specific activity of 0.078 units per milligram of protein. This is an order of magnitude greater than previous purifications of the plant enzyme.


Living on the edge: frontier voices.
Living on the edge is a universal lived experience. The humanbecoming school of thought is the theoretical underpinning described here to expand understanding of this phenomenon and contribute to nursing knowledge. A synthetic definition derived with concept inventing was created. The experience of living on the edge is risking venturing with sureness-unsureness surfacing with cherished engaging. At the theoretical level living on the edge is powering the originating of valuing connecting-separating.


Clinical factors related to recurrence after hepatic arterial concurrent chemoradiotherapy for advanced but liver-confined hepatocellular carcinoma.
Before the sorafenib era, advanced but liver-confined hepatocellular carcinoma (HCC) was treated by liver-directed therapy. Hepatic arterial concurrent chemoradiotherapy (CCRT) has been performed in our group, giving substantial local control but frequent failure. The aim of this study was to analyze patterns of failure and find out predictive clinical factors in HCC treated with a liver-directed therapy, CCRT. A retrospective analysis was done for 138 HCC patients treated with CCRT between May 2001 and November 2009. Protocol-based CCRT was performed with local radiotherapy (RT) and concurrent 5-fluorouracil (5-FU) hepatic arterial infusion chemotherapy (HAIC), followed by monthly HAIC (5-FU and cisplatin). Patterns of failure were categorized into three groups: infield, intrahepatic-outfield and extrahepatic failure. Treatment failure occurred in 34.0% of patients at 3 months after RT. Infield, intrahepatic-outfield and extrahepatic failure were observed in 12 (8.6%), 26 (18.7%) and 27 (19.6%) patients, respectively. Median progression-free survival for infield, outfield and extrahepatic failure was 22.4, 18 and 21.5 months, respectively. For infield failure, a history of pre-CCRT treatment was a significant factor (P = 0.020). Pre-CCRT levels of alpha-fetoprotein and prothrombin induced by vitamin K absence or antagonist-II were significant factors for extrahepatic failure (P = 0.029). Treatment failures after CCRT were frequent in HCC patients, and were more commonly intrahepatic-outfield and extrahepatic failures than infield failure. A history of pre-CCRT treatment and levels of pre-CCRT tumor markers were identified as risk factors that could predict treatment failure. More intensified treatment is required for patients presenting risk factors.


Crystal structure of the catalytic subunit of human protein phosphatase 1 and its complex with tungstate.
Protein phosphatase 1 (PP1) is a serine/threonine protein phosphatase that is essential in regulating diverse cellular processes. Here we report the crystal structure of the catalytic subunit of human PP1 gamma 1 and its complex with tungstate at 2.5 A resolution. The anomalous scattering from tungstate was used in a multiple wavelength anomalous dispersion experiment to derive crystallographic phase information. The protein adopts a single domain with a novel fold, distinct from that of the protein tyrosine phosphatases. A di-nuclear ion centre consisting of Mn2+ and Fe2+ is situated at the catalytic site that binds the phosphate moiety of the substrate. Proton-induced X-ray emission spectroscopy was used to identify the nature of the ions bound to the enzyme. The structural data indicate that dephosphorylation is catalysed in a single step by a metal-activated water molecule. This contrasts with other phosphatases, including protein tyrosine phosphatases, acid and alkaline phosphatases which form phosphoryl-enzyme intermediates. The structure of PP1 provides insight into the molecular mechanism for substrate recognition, enzyme regulation and inhibition of this enzyme by toxins and tumour promoters and a basis for understanding the expanding family of related phosphatases which include PP2A and PP2B (calcineurin).


Does cup-cage reconstruction with oversized cups provide initial stability in THA for osteoporotic acetabular fractures?
The incidence of acetabular fractures in osteoporotic patients is increasing. Immediate total hip arthroplasty (THA) has potential advantages, but achieving acetabular component stability is challenging and, at early followup, reported revision rates for loosening are high. This study measured acetabular component stability and the initial surface contact achieved between the acetabular component and unfractured region of the pelvis after THA using an oversized acetabular component and cup-cage reconstruction. Between November 2011 and November 2013, we treated 40 acute acetabular fractures in patients older than 70 years of age. Of these, 12 (30%) underwent immediate THA using an oversized acetabular component with screws inserted only into the ilium and a cup-cage construct. Postoperatively all patients were mobilized without weightbearing restrictions. Indications for immediate THA after acetabular fractures were displaced articular comminution deemed unreducible. Eleven of the 12 were prospectively studied to evaluate the initial stability of the reconstructions using radiostereometric analysis. One of the patients died of a pulmonary embolism after surgery, and the remaining 10 (median age, 81 years; range, 72-86 years) were studied. Of these, five were analyzed at 1 year and five were analyzed at 2 years. Acetabular component migration was defined as acceptable if less than the limits for primary THA that predict later loosening (1.76 mm of proximal migration and 2.53° of sagittal rotation). The contact surface between the acetabular component and ilium in direct continuity with the sacroiliac joint, and the ischium and pubis in direct continuity with the symphysis pubis, was measured on postoperative CT scans. At 1 year the median proximal migration was 0.83 mm (range, 0.09-5.13 mm) and sagittal rotation was 1.3° (range, 0.1°-7.4°). Three of the 10 components had migration above the suggested limits for primary THA at 1 year postoperatively. The contact surface achieved at surgery between the acetabular component and pelvis ranged from 11 to 17 cm(2) (15%-27% of each component). The majority of acetabular components in this cohort were stable despite the small contact surface achieved between the component and pelvic bone. Three of 10 migrated in excess of the limits that predict later loosening in primary THA but it remains to be seen whether these limits apply to this selected group of frail osteoporotic patients. We continue to use this technique routinely to treat patients with the same indications, but since the analysis of these data we have added screw fixation of the acetabular component to the ischial tuberosity and the superior pubic ramus. Level IV, therapeutic study.


Mechanisms of Soft Tissue Injury and Repair from Ballistics.
Projectile injuries to the face deserve particular attention to evaluate for involvement of critical structures and functional elements and treat the devastating effects on facial aesthetics. Ballistic trauma to the maxillofacial region often has significant soft tissue and bony defects, creating a greater challenge to the reconstructive surgeon. The main goals of treatment of projectile injuries to the face are decreasing the amount of contaminants and nonviable tissue at the site, ensuring functionality, and restoring aesthetic appearance. There exists a lack of robust research on the subject. Soft tissue injuries from high-velocity projectiles have changed the perspective on treating acute injuries to the face and neck. Injuries encountered during both Operation Iraqi Freedom and Operation Enduring Freedom give the author pause to reflect upon a different wounding pattern than that encountered in stateside trauma centers. Given the dissemination of high-velocity weaponry by enemy combatants such as the improvised explosive device, mortar round, and high-velocity rifles, a higher incidence of facial wounds has been reported. The mechanism of injury and a stepwise approach for surgical repair are discussed incorporating advanced trauma life support principles and a reconstructive ladder of repair.


Comparative Genomic and Secretomic Analysis Provide Insights Into Unique Agar Degradation Function of Marine Bacterium Vibrio fluvialis A8 Through Horizontal Gene Transfer.
Agarose-oligosaccharide production from agar degradation by agarase exhibits lots of advantages and good application prospects. In this study, a novel agar-degrading bacterium Vibrio sp. A8 was isolated from a red algae in the South China Sea. The whole genome sequencing with comparative genomic and secretomic analysis were used to better understand its genetic components about agar degradation. This strain exhibited good agarase production in artificial seawater after culture optimization. The complete genome (4.88 Mb) of this strain comprised two circular chromosomes (3.19 and 1.69 Mb) containing 4,572 protein-coding genes, 108 tRNA genes and 31 rRNA genes. This strain was identified as Vibrio fluvialis A8 by comparative genomic analysis based on genome phylogenetic tree and average nucleotide identity (ANI) similarity. Different from other 20 similar strains including three strains of the same species, V. fluvialis A8 possessed unique agar degradation ability with four β-agarases (GH50) and one α-1,3-L-NA2 hydrolase (GH117) due to the horizontal gene transfer. Secretomic analysis showed that only β-agarase (gene 3152) was abundantly expressed in the secretome of V. fluvialis A8. This agarase had a good substrate specificity and wide work conditions in complex environments, suggesting its potential application for agarose-oligosaccharide production.


HIV and tuberculosis in Durban, South Africa: adherence to two medication regimens.
Given that antiretroviral (ARV) medication adherence has been shown to be high in resource-limited countries, the question remains as to whether adherence will remain at that level as medications become more widely available. Comparing adherence to tuberculosis (TB) medications, which have been readily available, and ARV medications may help to indicate the likely future adherence to ARVs as access to these medications becomes more widespread. This study examined sense of coherence, social support, symptom status, quality of life, and adherence to medications in two samples of individuals being treated either for TB or human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) at clinics in Durban, South Africa. Findings revealed the distinctive socio-economic backgrounds of the two cohorts. Although there were significant differences with regard to the psychosocial variables, there were no significant differences by the two samples in adherence to medications as well as adherence to appointments. Given the self-selected nature of the participants in this study, namely those able to attend clinic, as well as those likely to be adherent to ARVs, there is every reason for caution in the interpretations of these findings. As access to ARV medications becomes more widely available in South Africa, the question remains as to whether such high adherence will be maintained given the constraints of access to food and other basic necessities.


Relationship of Lennox-Gastaut syndrome with perinatal event: A cross-sectional study.
About one-half of children with Lennox-Gastaut syndrome (LGS) have history of birth hypoxia or other perinatal event but the knowledge about clinical, radiological profile and severity of epilepsy in these children as compared to those without a perinatal event is not known. Thirty-one children with LGS were enrolled in this study and divided into two groups: One group with the perinatal event and other group without evidence of the perinatal event. We hypothesized that LGS with the perinatal event will have an early age of onset of LGS, more motor deficits and abnormal brain magnetic resonance imaging (MRI) and more severe epilepsy. There were 17 children in the perinatal event group and 14 in the other group. The mean age of onset of illness was significantly earlier in the perinatal event group (P < 0.05). More children in the perinatal event group had delayed milestones (P < 0.05), had higher seizure frequency (P < 0.05) however; there was no significant difference in number of anti-epileptic drugs consumed, motor deficits or MRI abnormalities. LGS children with the perinatal event have more severe epilepsy with early onset of disease and delayed milestones. History of perinatal insult in these children may help in predicting prognosis in LGS.


Pleuropulmonary and lymph node progression after docetaxel - benefits from treatment with cabazitaxel in metastatic prostate cancer.
To date, there are no guidelines for a rational and more favourable sequence of treatment after docetaxel. Two drugs (cabazitaxel and abiraterone) have recently been approved as second-line treatment after docetaxel failure in metastatic castration-resistant prostate cancer (mCRPC), but there are no studies comparing abiraterone versus cabazitaxel. The most suitable drug is chosen based on the physician's opinion and the patient's characteristics. In patients with a good performance status who are able to receive either treatment, it would be convenient to begin with cabazitaxel and to reserve abiraterone in case there is a worsening of the general status, in consideration of abiraterone's more favourable toxicity profile. We describe the case of a 74-year-old male with mCRPC who presented with an interesting and uncommon tumour dissemination (pleuropulmonary) occurring after the first standard treatment with docetaxel. Intravenous treatment with cabazitaxel 25 mg/m(2) and oral prednisone 10 mg continuously was initiated. The patient received a total of 8 cycles of chemotherapy. A reduction of mediastinal adenopathies and infrarenal para-aortic stable bone involvement and an absence of pleural effusion were observed. No relevant toxicity was noted. Since February 2012, a progressive PSA increase without clinical deterioration has been noted. The selection criteria for second- and third-line systemic treatment and the excellent response obtained with cabazitaxel in an unusual disease setting are described. The results confirm the long duration and quality of response of cabazitaxel treatment. Further therapeutic options in this group of patients are suggested.


Cardiac arrhythmias and cardiac arrest related to mushroom poisoning: A case report.
Mushroom exposure is a global health issue. The manifestations of mushroom poisoning (MP) may vary. Some species have been reported as rhabdomyolytic, hallucinogenic, or gastrointestinal poisons. Critical or even fatal MPs are mostly attributable to Amanita phalloides, with the development of severe liver or renal failure. Myocardial injury and even cases mimicking ST-segment elevation myocardial infarction (STEMI) have been previously reported, while cardiac arrhythmia or cardiac arrest is not commonly seen. We report a 68-year-old woman with MP who suffered from delirium, seizure, long QT syndrome on electrocardiogram (ECG), severe cardiac arrhythmias of multiple origins, and cardiac arrest. She was intubated and put on blood perfusion. Her kidney and liver functions were intact; creatine kinase-MB was mildly elevated, and then fell within normal range during her hospital stay. We sent the mushrooms she left for translation elongation factor subunit 1α, ribosomal RNA gene sequence, and internal transcribed spacer sequence analyses. There were four kinds of mushrooms identified, two of which were found to be toxic. This is the first time that we found cardiac toxicity caused by Panaeolus subbalteatus and Conocybe lactea, which were believed to be toxic to the liver, kidney, and brain. We suggest that intensive monitoring and ECG follow-up are essential to diagnose prolonged QT interval and different forms of tachycardia in MP patients, even without the development of severe liver or renal failure. The mechanisms need to be further investigated and clarified based on animal experiments and molecular signal pathways.


Did increased availability of pediatric palliative care lead to improved palliative care outcomes in children with cancer?
Awareness for pediatric palliative care in children with cancer increased in the last ten years in Germany. In this study we sought to determine whether this change in awareness led to improved palliative care outcomes in children dying due to cancer. In 2005 we interviewed a cohort of 48 bereaved parents who had a lost a child to cancer approximately five years earlier (2000 cohort), and in 2010 we interviewed another cohort of 48 parents who had lost a child due to cancer approximately five years before (2005 cohort). Children of the 2000 cohort were cared for by six specialized oncology departments in North-Rhine-Westphalia (NRW), Germany, and children of the 2005 cohort by 16 specialized pediatric oncology departments in NRW, Germany. Parents of both cohorts were interviewed using the Survey of Caring for Children with Cancer (SCCC). The children of both parental cohorts were similar in terms of disease characteristics and sociodemographic variables. Children suffered in a very similar manner from core symptoms such as pain and dyspnea. However, symptom treatment increased for all symptoms. In case of treatment of anxiety the increase was statistically significant (p=0.035). Location of care changed with almost three-quarters of the 2005 cohort receiving palliative home care, significantly more than in the 2000 cohort (p=0.007). Additionally, fewer children of the 2005 cohort died in the intensive care unit. While the location of care during the end-of-life period shifted (from hospital to home), there remains substantial work to ease the suffering in children with cancer at end of life.


The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast.
Intracellular trafficking pathways control residency and bioactivity of integral membrane proteins at the cell surface. Upon internalisation, surface cargo proteins can be delivered back to the plasma membrane via endosomal recycling pathways. Recycling is thought to be controlled at the metabolic and transcriptional level, but such mechanisms are not fully understood. In yeast, recycling of surface proteins can be triggered by cargo deubiquitination and a series of molecular factors have been implicated in this trafficking. In this study, we follow up on the observation that many subunits of the Rpd3 lysine deacetylase complex are required for recycling. We validate ten Rpd3-complex subunits in recycling using two distinct assays and developed tools to quantify both. Fluorescently labelled Rpd3 localises to the nucleus and complements recycling defects, which we hypothesised were mediated by modulated expression of Rpd3 target gene(s). Bioinformatics implicated 32 candidates that function downstream of Rpd3, which were over-expressed and assessed for capacity to suppress recycling defects of rpd3∆ cells. This effort yielded three hits: Sit4, Dit1 and Ldb7, which were validated with a lipid dye recycling assay. Additionally, the essential phosphatidylinositol-4-kinase Pik1 was shown to have a role in recycling. We propose recycling is governed by Rpd3 at the transcriptional level via multiple downstream target genes.


Biosynthesis of UDP-xylose. Cloning and characterization of a novel Arabidopsis gene family, UXS, encoding soluble and putative membrane-bound UDP-glucuronic acid decarboxylase isoforms.
UDP-xylose (Xyl) is an important sugar donor for the synthesis of glycoproteins, polysaccharides, various metabolites, and oligosaccharides in animals, plants, fungi, and bacteria. UDP-Xyl also feedback inhibits upstream enzymes (UDP-glucose [Glc] dehydrogenase, UDP-Glc pyrophosphorylase, and UDP-GlcA decarboxylase) and is involved in its own synthesis and the synthesis of UDP-arabinose. In plants, biosynthesis of UDP-Xyl is catalyzed by different membrane-bound and soluble UDP-GlcA decarboxylase (UDP-GlcA-DC) isozymes, all of which convert UDP-GlcA to UDP-Xyl. Because synthesis of UDP-Xyl occurs both in the cytosol and in membranes, it is not known which source of UDP-Xyl the different Golgi-localized xylosyltransferases are utilizing. Here, we describe the identification of several distinct Arabidopsis genes (named AtUXS for UDP-Xyl synthase) that encode functional UDP-GlcA-DC isoforms. The Arabidopsis genome contains five UXS genes and their protein products can be subdivided into three isozyme classes (A-C), one soluble and two distinct putative membrane bound. AtUxs from each class, when expressed in Escherichia coli, generate active UDP-GlcA-DC that converts UDP-GlcA to UDP-Xyl. Members of this gene family have a large conserved C-terminal catalytic domain (approximately 300 amino acids long) and an N-terminal variable domain differing in sequence and size (30-120 amino acids long). Isoforms of class A and B appear to encode putative type II membrane proteins with their catalytic domains facing the lumen (like Golgi-glycosyltransferases) and their N-terminal variable domain facing the cytosol. Uxs class C is likely a cytosolic isoform. The characteristics of the plant Uxs support the hypothesis that unique UDP-GlcA-DCs with distinct subcellular localizations are required for specific xylosylation events.


"Recovering, not recovered" Hospital disaster resilience: a case-study from the 2015 earthquake in Nepal.
Disasters are an increasing threat to human health, but we know little about their impact on health services, particularly in low and middle-income settings. 'Resilient hospitals' have been increasingly recognized as a cornerstone of disaster management. While various frameworks of hospital resilience exist, they emerged from pre-disaster considerations, and do not incorporate evidence from post-disaster settings. This dissertation investigated the impact of a large-scale sudden onset disaster in a tertiary hospital in Nepal, and explored its resilience mechanisms. This consists of an in-depth case-study combining quantitative data from routinely generated hospital records and qualitative data from semi-structured interviews with hospital staff. We used both advanced statistical methods and mixed inductive and deductive coding to analyze the data. Most of the admitted earthquake victims required surgical interventions and long hospitalizations, considerably straining the hospital. For six weeks, the average number of daily admissions decreased. During this period, the share of injury-related admissions was particularly high, and such admissions were particularly long compared to the baseline. Admissions due to other conditions relatively decreased and were shorter. We found that the hospital's resilience was highly dependent on emerging adaptations, in addition to the pre-existing disaster plan. Individual resilience of staff also played a major role, and was influenced by senses of safety, meaningfulness, and belonging. Hospitals should prepare resources and plan for their known disaster risks, but should also allow for a certain flexibility to innovative adaptions to emerging, unforeseen challenges. Challenges faced by hospital workers should not be undermined, and addressing them will increase hospital resilience.


Variant muscle fibers connecting the orbicularis oculi to the orbicularis oris: case report.
The orbicularis oculi (OOc) is a sphincteric muscle of the eyelids, whereas contraction of the orbicularis oris (OOr), another sphincteric muscle, causes narrowing of the lips. Facial muscle fibers normally blend with adjacent muscles. However, muscle fibers connecting the various facial muscles that have different actions and that are located at distant sites, such as the OOc and the OOr have been rarely reported. Herein, we report a rare case of connecting fibers between the inferior margin of the OOc and the OOr. These connecting fibers were blended with the OOr between the inserting fibers of the levator labii superioris and levator anguli oris. Contraction of such variant muscles might affect typical facial expressions.


Mammalian STE20-Like Kinase 2 Promotes Lipopolysaccharides-Mediated Cardiomyocyte Inflammation and Apoptosis by Enhancing Mitochondrial Fission.
In this study, we analyzed the role of mammalian STE20-like protein kinase 2 (Mst2), a serine-threonine protein kinase, in Lipopolysaccharides (LPS)-mediated inflammation and apoptosis in the H9C2 cardiomyocytes. Mst2 mRNA and protein levels were significantly upregulated in the LPS-treated H9C2 cardiomyocytes. LPS treatment induced expression of IL-2, IL-8, and MMP9 mRNA and proteins in the H9C2 cardiomyocytes, and this was accompanied by increased caspase-3/9 mediating H9C2 cardiomyocyte apoptosis. LPS treatment also increased mitochondrial reactive oxygen species (ROS) and the levels of antioxidant enzymes, such as GSH, SOD, and GPX, in the H9C2 cardiomyocytes. The LPS-treated H9C2 cardiomyocytes showed lower cellular ATP levels and mitochondrial state-3/4 respiration but increased mitochondrial fragmentation, including upregulation of the mitochondrial fission genes Drp1, Mff, and Fis1. LPS-induced inflammation, mitochondrial ROS, mitochondrial fission, and apoptosis were all significantly suppressed by pre-treating the H9C2 cardiomyocytes with the Mst2 inhibitor, XMU-MP1. However, the beneficial effects of Mst2 inhibition by XMU-MP1 were abolished by carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP), a potent activator of mitochondrial fission. These findings demonstrate that Mst2 mediates LPS-induced cardiomyocyte inflammation and apoptosis by increasing mitochondrial fission.


Complete Androgen Insensitivity Syndrome: A Problem-Based Learning Case.
To address the lack of medical education on lesbian, gay, bisexual, transgender, queer (LGBTQ), and difference in sex development-affected (DSD-affected) individuals, the University of Louisville initiated the eQuality project. The mission of eQuality is to promote mastery of the skills, knowledge, and attitudes required for excellent care for patients within the LGBTQ, gender-nonconforming, and DSD community. This problem-based learning (PBL) case was implemented to address both medical and psychosocial factors that affect individuals born with DSD and their families. Students complete a 10-question multiple-choice pretest prior to distribution of the PBL case and a 10-question multiple-choice posttest after completion of the PBL. They also evaluate the efficacy of the case by answering three open-ended questions regarding comprehension, competence, and attitudes, as well as any barriers to acquisition and implementation of knowledge. Along with a specific lecture and patient panel, this piece helps to incorporate more information specifically on DSD into the curriculum. The posttest average score was 20% higher than the pretest. The posttest (n = 144) had a range of 90% to 99% of students answering each question correctly, whereas the range for the pretest (n = 155) was from 29% to 94%. Based on the objective assessment and student comments, learning increased through this PBL process. Despite this clear increase in knowledge, some students believed that either societal barriers or personal barriers would prevent them from implementing their newly gained knowledge in practice.


DNA damage response is suppressed by the high cyclin-dependent kinase 1 activity in mitotic mammalian cells.
DNA damage response (DDR) is vital for genomic stability, and its deficiency is linked to tumorigenesis. Extensive studies in interphase (G(1)-S-G(2)) mammalian cells have revealed the mechanisms of DDR in great detail; however, how mitotic cells respond to DNA damage remains less defined. We report here that a full DDR is suppressed in mitotic mammalian cells until telophase/cytokinesis. Although early DDR markers such as the phosphorylations of ataxia telangiectasia mutated (ATM) and histone H2A.x (H2AX) can be readily detected, the ionizing radiation-induced foci (IRIF) formation of late DDR markers such as breast cancer type 1 susceptibility protein (BRCA1) and p53-binding protein 1 (53BP1) are absent until the telophase/cytokinesis stage. We further showed that the IR-induced ubiquitination cascade around DNA damage sites did not occur in mitotic cells, which explains, at least in part, why BRCA1 and 53BP1 cannot be recruited to the damaged sites. These observations indicate that DDR is suppressed in mitotic cells after the step of γH2AX formation. Not surprisingly, we found that the absence of a full DDR in mitotic cells was associated with the high cyclin-dependent kinase 1 (CDK1) activities. More 53BP1 IRIF could be detected when the irradiated mitotic cells were treated with a CDK1 inhibitor. Further, the activation of CDK5 in interphase cells impedes the formation of 53BP1 IRIF. Together, these results suggest that the DDR is suppressed by the high CDK1 activity in mitotic mammalian cells.


Cellular and transcriptomic analysis of human mesenchymal stem cell response to plasma-activated hydroxyapatite coating.
Atmospheric pressure plasma has recently emerged as a technique with a promising future in the medical field. In this work we used the technique as a post-deposition modification process as a means to activate hydroxyapatite (HA) coatings. Contact angle goniometry, optical profilometry, scanning electron microscopy morphology imaging and X-ray photoelectron spectroscopy analysis demonstrate that surface wettability is improved after treatment, without inducing any concomitant damage to the coating. The protein adsorption pattern has been found to be preferable for MSC, and this may result in greater cell attachment and adhesion to plasma-activated HA than to untreated samples. Cell cycle distribution analysis using flow cytometry reveals a faster transition from G(1) to S phase, thus leading to a faster cell proliferation rate on plasma-activated HA. This indicates that the improvement in surface wettability independently enhances cell attachment and cell proliferation, which is possibly mediated by FAK phosphorylation. Pathway-specific polymerase chain reaction arrays revealed that wettability has a substantial influence on gene expression during osteogenic differentiation of human MSC. Plasma-activated HA tends to enhance this process by systemically deregulating multiple genes. In addition, the majority of these deregulated genes had been appropriately translated, as confirmed by ELISA protein quantification. Lastly, alizarin red staining showed that plasma-activated HA is capable of improving mineralization for up to 3 weeks of in vitro culture. It was concluded from this study that atmospheric pressure plasma is a potent tool for modifying the biological function of a material without causing thermal damage, such that adhesion molecules and drugs might be deposited on the original coating to improve performance.


Socioeconomic and air pollution correlates of adult asthma, heart attack, and stroke risks in the United States, 2010-2013.
Asthma in the United States has become an important public health issue, with many physicians, regulators, and scientists elsewhere expressing concern that criterion air pollutants have contributed to a rising tide of asthma cases and symptoms. This paper studies recent associations (from 2008 to 2012) between self-reported asthma experiences and potential predictors, including age, sex, income, education, smoking, and county-level average annual ambient concentrations of ozone (O3) and fine particulate matter (PM2.5) levels recorded by the U.S. Environmental Protection Agency, for adults 50 years old or older for whom survey data are available from the Centers for Disease Control and Prevention (CDC) Behavioral Risk Factor Surveillance System (BRFSS). We also examine associations between these variables and self-reported heart attack and stroke experience; all three health outcomes are positively associated with each other. Young divorced women with low incomes are at greatest risk of asthma, especially if they are ever-smokers. Income is an important confounder of other relations. For example, in logistic regression modeling, PM2.5 is positively associated (p<0.06) with both stroke risk and heart attack risk when these are regressed only against PM2.5, sex, age, and ever-smoking status, but not when they are regressed against these variables and income. In this data set, PM2.5 is significantly negatively associated with asthma risk in regression models, with a 10μg/m3 decrease in PM2.5 corresponding to about a 6% increase in the probability of asthma, possibly because of confounding by smoking, which is negatively associated with PM2.5 and positively associated with asthma risk. A variety of non-parametric methods are used to quantify these associations and to explore potential causal interpretations.


Effect of smoking on physical function in older adults: a Mendelian randomisation analysis.
Smoking is widely known to be damaging to health. It greatly increases the risk of various cancers, cardiovascular and respiratory diseases, and remains the leading preventable cause of morbidity and mortality in the UK. The article tested the hypothesis that smoking behaviour (either smoking or non-smoking) is associated with physical function, and assessed inferred causality using genetic predisposition to smoking behaviour as an instrumental variable. Data were drawn from the English Longitudinal Study of Ageing, waves 1-9 (mean age 65.8 years). Physical function was assessed by means of the body mobility and the activity of daily living indices. Polygenic scores for smoking behaviour were used as instrumental variables in a Mendelian randomisation framework. Instrumental variable estimators were used to examine causal effects. Among UK older adults (n=29 139), impaired physical function was significantly higher in (current) smokers compared with non-smokers. Relatively to non-smokers, smokers reported a higher level of impairment both in the body mobility index (β=5.553; 95% CI 1.029 to 10.077) and in the activities of daily living index (β=1.908; 95% CI 0.196 to 3.619). This study demonstrates smoking behaviour to be a potential causal risk factor for physical function during ageing in the UK population. Accordingly, the benefits of smoking cessations may extend to physical function.


Slow molecular dynamics of water in a lyotropic complex fluid studied by deuterium conventional and spin-lattice relaxometry NMR.
A nuclear magnetic resonance study of protons and deuterons in the mesomorphic phases of the micellar lyotropic mixture potassium laurate/1-decanol/heavy water is reported. The slow dynamical behavior of water molecules has been investigated with deuterons spin-lattice relaxation dispersion in the Larmor frequency range 10(3)<nu(L)<4.2x10(7) Hz. In order to compare relative behaviors additional T1 dispersion of micellar protons has been measured in the same compound, temperature, and Larmor frequency range. From the experimental behaviors, we conclude that in the nematic phases the water slow reorientational dynamics is closely related to the slow reorientation of the micellar aggregates. In addition, conventional deuterium nuclear magnetic resonance at nu(L)=4.2x10(7) Hz spectra has been measured at different places in the phase diagram. The line shapes show a quadrupolar splitting in nematic phases, meanwhile in the isotropic phase the spectral structure collapses in a single line. This indicates that in the nematic phases the water reorientations are not enough to average the deuterons quadrupolar Hamiltonian. On the other hand, fast isotropic water reorientations reduce the quadrupolar interactions in the isotropic phase.


Rescued tolerant CD8 T cells are preprogrammed to reestablish the tolerant state.
Tolerant self-antigen-specific CD8 T cells fail to proliferate in response to antigen, thereby preventing autoimmune disease. By using an in vivo mouse model, we show that tolerant T cells proliferate and become functional under lymphopenic conditions, even in a tolerogenic environment. However, T cell rescue is only transient, with tolerance reimposed upon lymphorepletion even in the absence of tolerogen (self-antigen), challenging the prevailing paradigm that continuous antigen exposure is critical to maintain tolerance. Genome-wide messenger RNA and microRNA profiling revealed that tolerant T cells have a tolerance-specific gene profile that can be temporarily overridden under lymphopenic conditions but is inevitably reimposed, which suggests epigenetic regulation. These insights into the regulatory mechanisms that maintain or break self-tolerance may lead to new strategies for the treatment of cancer and autoimmunity.