Activation of alternative NF-κB signaling during recovery of disuse-induced loss of muscle oxidative phenotype.
Physical inactivity-induced loss of skeletal muscle oxidative phenotype (OXPHEN), often observed in chronic disease, adversely affects physical functioning and quality of life. Potential therapeutic targets remain to be identified, since the molecular mechanisms involved in reloading-induced recovery of muscle OXPHEN remain incompletely understood. We hypothesized a role for alternative NF-κB, as a recently identified positive regulator of muscle OXPHEN, in reloading-induced alterations in muscle OXPHEN. Markers and regulators (including alternative NF-κB signaling) of muscle OXPHEN were investigated in gastrocnemius muscle of mice subjected to a hindlimb suspension/reloading (HLS/RL) protocol. Expression levels of oxidative phosphorylation subunits and slow myosin heavy chain isoforms I and IIA increased rapidly upon RL. After an initial decrease upon HLS, mRNA levels of peroxisome proliferator-activated receptor (PPAR)-γ coactivator (PGC) molecules PGC-1α and PGC-1β and mRNA levels of mitochondrial transcription factor A (Tfam) and estrogen-related receptor α increased upon RL. PPAR-δ, nuclear respiratory factor 1 (NRF-1), NRF-2α, and sirtuin 1 mRNA levels increased during RL although expression levels were unaltered upon HLS. In addition, both Tfam and NRF-1 protein levels increased significantly during the RL period. Moreover, upon RL, IKK-α mRNA and protein levels increased, and phosphorylation of P100 and subsequent processing to P52 were elevated, reflecting alternative NF-κB activation. We conclude that RL-induced recovery of muscle OXPHEN is associated with activation of alternative NF-κB signaling.


Refolding of the non-specific neutral protease from Bacillus stearothermophilus proceeds via an autoproteolytically sensitive intermediate.
A very thermostable variant of the thermolysin-like protease from Bacillus stearothermophilus (G8C/N60C) was previously created by introduction of a disulfide bond into the cysteine-free pseudo-wild type variant (pWT) and thus fixing the unfolding region 56-69. In the present paper, we show that G8C/N60C and pWT can be reactivated from the completely unfolded states, accessible at >or=7.5M guanidine hydrochloride, and analyze the kinetics of folding, autoproteolytic degradation and aggregation. From changes in the fluorescence spectra with time of renaturation, it can be concluded that a folding intermediate with native-like structure, but which is still inactive and sensitive to autoproteolysis, is rapidly formed after renaturation has initiated. The critical region 56-69 of pWT is involved in the autoproteolytic sensitivity of the intermediate as we conclude from the differences in the chevron plots of the first-order rate constants of reactivation and the fragmentation patterns in SDS-PAGE of pWT and G8C/N60C.


Are mice calorically restricted in nature?
An important question about traditional caloric restriction (CR) experiments on laboratory mice is how food intake in the laboratory compares with that of wild mice in nature. Such knowledge would allow us to distinguish between two opposing views of the anti-aging effect of CR--whether CR represents, in laboratory animals, a return to a more normal level of food intake, compared with excess food consumption typical of laboratory conditions or whether CR represents restriction below that of animals living in nature, i.e. the conditions under which house mice evolved. To address this issue, we compared energy use of three mouse genotypes: (1) laboratory-selected mouse strains (= laboratory mice), (2) house mice that were four generations or fewer removed from the wild (= wild-derived mice) and (3) mice living in nature (= wild mice). We found, after correcting for body mass, that ad libitum fed laboratory mice eat no more than wild mice. In fact, under demanding natural conditions, wild mice eat even more than ad libitum fed laboratory mice. Laboratory mice do, however, eat more than wild-derived mice housed in similar captive conditions. Therefore, laboratory mice have been selected during the course of domestication for increased food intake compared with captive wild mice, but they are not particularly gluttonous compared with wild mice in nature. We conclude that CR experiments do in fact restrict energy consumption beyond that typically experienced by mice in nature. Therefore, the retarded aging observed with CR is not due to eliminating the detrimental effects of overeating.


The relationship of agarose gel structure to the sieving of spheres during agarose gel electrophoresis.
To understand the organization of fibers in an agarose gel, digitized electron micrographs are used here to determine the frequency distribution of interfiber distance (2Pc) in thin sections of agarose gels. For a preparation of underivatized agarose, a 1.5% gel has a Pc distribution that is indistinguishable from the Pc distribution of a computer-generated, random-fiber gel; the log of the occurrence frequency (F) decreases linearly as a function of Pc. As the agarose concentration decreases below 1.5%, the semilogarithmic F versus Pc plot becomes progressively less linear. Two straight lines represent the data; the plot is steeper at the lower Pc values. As the percentage of agarose increases above 1.5%, the semilogarithmic F versus Pc plot becomes steeper at the higher Pc values. This change in the shape of semilogarithmic F versus Pc plots is possibly explained by the existence in agarose gels of two zones, one whose Pc distribution is more sensitive to the average agarose concentration than the other. To compare the structure of agarose gels to their sieving during electrophoresis, the root mean square value of Pc (Pc) is compared to the sieving-based radius of the effective pore (PE; Griess et. al. (16)) for both underivatized agarose and a derivatized agarose that has a smaller PE at any given agarose percentage. For 0.8-2.0% gels of either underivatized or derivatized agarose, PE/Pc is a constant within experimental error. Deviations from this constant are observed at lower gel percentages. This relationship of PE to Pc constrains theoretical descriptions of the motion of spheres in fibrous networks.


Intrinsic disorder controls two functionally distinct dimers of the master transcription factor PU.1.
Transcription factors comprise a major reservoir of conformational disorder in the eukaryotic proteome. The hematopoietic master regulator PU.1 presents a well-defined model of the most common configuration of intrinsically disordered regions (IDRs) in transcription factors. We report that the structured DNA binding domain (DBD) of PU.1 regulates gene expression via antagonistic dimeric states that are reciprocally controlled by cognate DNA on the one hand and by its proximal anionic IDR on the other. The two conformers are mediated by distinct regions of the DBD without structured contributions from the tethered IDRs. Unlike DNA-bound complexes, the unbound dimer is markedly destabilized. Dimerization without DNA is promoted by progressive phosphomimetic substitutions of IDR residues that are phosphorylated in immune activation and stimulated by anionic crowding agents. These results suggest a previously unidentified, nonstructural role for charged IDRs in conformational control by mitigating electrostatic penalties that would mask the interactions of highly cationic DBDs.


Photoplethysmography and its application in clinical physiological measurement.
Photoplethysmography (PPG) is a simple and low-cost optical technique that can be used to detect blood volume changes in the microvascular bed of tissue. It is often used non-invasively to make measurements at the skin surface. The PPG waveform comprises a pulsatile ('AC') physiological waveform attributed to cardiac synchronous changes in the blood volume with each heart beat, and is superimposed on a slowly varying ('DC') baseline with various lower frequency components attributed to respiration, sympathetic nervous system activity and thermoregulation. Although the origins of the components of the PPG signal are not fully understood, it is generally accepted that they can provide valuable information about the cardiovascular system. There has been a resurgence of interest in the technique in recent years, driven by the demand for low cost, simple and portable technology for the primary care and community based clinical settings, the wide availability of low cost and small semiconductor components, and the advancement of computer-based pulse wave analysis techniques. The PPG technology has been used in a wide range of commercially available medical devices for measuring oxygen saturation, blood pressure and cardiac output, assessing autonomic function and also detecting peripheral vascular disease. The introductory sections of the topical review describe the basic principle of operation and interaction of light with tissue, early and recent history of PPG, instrumentation, measurement protocol, and pulse wave analysis. The review then focuses on the applications of PPG in clinical physiological measurements, including clinical physiological monitoring, vascular assessment and autonomic function.


Carbon payments can cost-effectively improve logging sustainability in the Amazon.
Selective logging is pervasive across the tropics and unsustainable logging depletes forest biodiversity and carbon stocks. Improving the sustainability of logging will be crucial for meeting climate targets. Carbon-based payment for ecosystem service schemes, including REDD+, give economic value to standing forests and can protect them from degradation, but only if the revenue from carbon payments is greater than the opportunity cost of forgone or reduced logging. We currently lack understanding of whether carbon payments are feasible for protecting Amazonian forests from logging, despite the Amazon holding the largest unexploited timber reserves and an expanding logging sector. Using financial data and inventories of >660,000 trees covering 52,000 ha of Brazilian forest concessions, we estimate the carbon price required to protect forests from logging. We estimate that a carbon price of $7.90 per tCO2 is sufficient to match the opportunity costs of all logging and fund protection of primary forest. Alternatively, improving the sustainability of logging operations by ensuring a greater proportion of trees are left uncut requires only slightly higher investments of $7.97-10.45 per tCO2. These prices fall well below the current compliance market rate and demonstrate a cost-effective opportunity to safeguard large tracts of the Amazon rainforest from further degradation.


Studies on the structure of sphingomyelinase. Amino acid composition and heterogeneity on isoelectric focusing.
Sphingomyelinase, purified to apparent homogeneity from human placenta, is an acidic protein, as judged from its amino acid composition and by isoelectric focusing of the carboxymethylated protein. The amino acid composition is characterized by an approximately equal content of hydrophobic and polar amino acid residues. The reduced-alkylated polypeptides were separated into two groups. Most of the polypeptides were heterogeneous with pI values of 4.4-5.0, but an additional more minor component was observed at pI 5.4. Liquid isoelectric focusing resolved the purified enzyme into a single major component (pI 4.7-4.8), a minor component (pI 5.0-5.4) and a plateau region of activity (pI 6-7). On thin-layer isoelectric focusing, the protein profile obtained from each of these regions was the same. In addition, the substrate specificity, Km values and effect of inhibitory substances were identical. We conclude that sphingomyelinase is an acidic, microheterogeneous protein that likely exists as a holopolymer of a single major polypeptide chain. the heterogeneity of the intact protein on isoelectric focusing appears to reflect this microheterogeneity, which is influenced by a tendency to associate with itself and with detergents such as Triton X-100.


Blueshift in Trifurcated Hydrogen Bonds: A Tradeoff between Tetrel Bonding and Steric Repulsion.
We have quantum chemically investigated the origin of the atypical blueshift of the H-C bond stretching frequency in the hydrogen-bonded complex X- •••H3 C-Y (X, Y=F, Cl, Br, I), as compared to the corresponding redshift occurring in Cl- •••H3 N and Cl- •••H3 C-H, using relativistic density functional theory (DFT) at ZORA-BLYP-D3(BJ)/QZ4P. Previously, this blueshift was attributed, among others, to the contraction of the H-C bonds as the H3 C moiety becomes less pyramidal. Herein, we provide quantitative evidence that, instead, the blueshift arises from a direct and strong X- •••C interaction of the HOMO of A- with the backside lobe on carbon of the low-lying C-Y antibonding σ* LUMO of the H3 C-Y fragment. This X- •••C bond, in essence a tetrel bond, pushes the H atoms towards a shorter H-C distance and makes the H3 C moiety more planar. The blueshift may, therefore, serve as a diagnostic for tetrel bonding.


Effects of pH and manure on transport of sulfonamide antibiotics in soil.
Sulfonamide antibiotics are a commonly used group of compounds in animal husbandry. They are excreted with manure, which is collected in a storage lagoon in certain types of confined animal feeding operations. Flood irrigation of forage fields with this liquid manure creates the potential risk of groundwater contamination in areas with shallow groundwater levels. We tested the hypothesis that-in addition to the soil characteristics-manure as cosolute and manure pH are two major parameters influencing sulfonamide transport in soils. Solute displacement experiments in repacked, saturated soil columns were performed with soil (loamy sand) and manure from a dairy farm in California. Breakthrough of nonreactive tracer and sulfadimethoxine, sulfamethazine, and sulfamethoxazole at different solution pH (5, 6.5, 8.5) with and without manure was modeled using Hydrus-1D to infer transport and reaction parameters. Tracer and sulfonamide breakthrough curves were well explained by a model concept based on physical nonequilibrium transport, equilibrium sorption, and first-order dissipation kinetics. Sorption of the antibiotics was low ( K₄ ≤ 0.7 L kg) and only weakly influenced by pH and manure. However, sulfonamide attenuation was significantly affected by both pH and manure. The mass recovery of sulfonamides decreased with decreasing pH, e.g., for sulfamethoxazole from 77 (pH 8.5) to 56% (pH 5). The sulfonamides were highly mobile under the studied conditions, but manure application increased their attenuation substantially. The observed attenuation was most likely caused by a combination of microbial transformation and irreversible sorption to the soil matrix.


Monitoring liver safety in drug development: the GSK experience.
To promptly identify and evaluate liver safety events, an evidence-based liver safety system was created for global Phase I-III clinical trials. The goals of this system included improving clinical trial subject safety, expanding information on liver safety events, and improving data quality across studies by establishing and communicating: Two different algorithms for liver stopping criteria were developed. The most stringent criteria were selected for healthy volunteers in Phase I studies, where no treatment benefit is anticipated and clinical safety data are limited. With an interest in assessing potential liver "tolerance" or adaptation with accruing safety information, slightly higher liver chemistry thresholds were set for Phase II-III studies. This paper will describe the importance of liver safety in drug development, laboratory tests used to monitor liver safety, the rationale for selected liver chemistry subject stopping criteria, and implementation of this safety system.


The effect of CPP-ACP and Nd:YAG laser on the bond strength of softened dentin.
The purpose of this study was to investigate the effect of CPP-ACP treatment and Nd:YAG laser on microtensile bond strength (µTBS) of softened dentin. Sixty samples were obtained from thirty sound third molars. All samples were submitted to dentin softening procedure, by the immersion of the specimens in 30 mL of Sprite Zero for 30 min. Afterwards, the samples were randomly divided according to the CPP-ACP treatment: CG-Control group; MP-treated with CPP-ACP paste (MI Paste); MPP-treated with CPP-ACP+900 ppm NaF paste (MI Paste Plus). Each group was further divided according to bonding procedure: NL-No laser; L-Laser irradiation after adhesive application and before polymerization. The laser parameters used were 1.4 W, 10 Hz, 140 mJ/pulse, with an optic fiber of 320 µm, generating energy of 174 J/cm(2) per pulse. All samples were restored with Clearfil SE Bond/Filtek Z350 XT. After 24 h, the restored samples were cut into beams (± 1 mm(2) adhesive interface area) and subjected to a µTBS test. Data were analyzed by two-way ANOVA test and Holm-Sidak post-hoc method (α = 0.05). The treatment with CPP-ACP pastes did not significantly affect softened dentin µTBS (p = 0.070). Statistic revealed significant reduction on µTBS values for CG/L, leading to the rejection of the second null hypothesis (p < 0.001). Both CPP-ACP based pastes did not affect µTBS of softened dentin for the adhesive system utilized. The Nd:YAG laser irradiation after application of adhesive system did affect µTBS values of softened dentin samples untreated with CPP-ACP based pastes.


[Spatial and temporal distribution of physicochemical features in the habitat of whale shark Rhincodon typus (Orectolobiformes: Rhincodontidae) in the north of Mexican Caribbean].
Large groups of whale sharks (Rhincodon typus) are common in the North Mexican Caribbean and gather between May and September each year. We describe their spatial and temporal distribution, and the physicochemical (temperature, dissolved oxygen, light extinction coefficient and salinity) and biological conditions during the presence and absence seasons of R. typus. A total of 26 sampling stations were monitored to record whale shark sightings and physicochemical variables during 10 field campaigns from April 2005 to March 2006. At each station, zooplankton and water samples, for chlorophyll-a and nutrients determination, were collected. Physicochemical conditions were significantly different between presence-absence seasons (ANOSIM, Rglobal = 0.632). The R. typus season was characterized by low salinity values, and higher temperature, chlorophyll-a, dissolved oxygen and inorganic nitrogen concentrations values. Average zooplankton biomass was lower during the absence season, while recorded the maximum values during the presence one. Furthermore, these values were also observed in areas with higher species abundances, supporting the hypothesis that the site is used by R. typus primarily as a foraging area. We conclude that physicochemical conditions of the study area promote the biological productivity, which explains the spatial and temporal variability of R. typus.


HMGB-1 as a target for inflammation controlling.
In the present study, we evaluated recent patents that describe products or methods able to down-regulate the pro-inflammatory action of HMGB-1, also called as amphoterin. High Mobility Group Box-1 (HMGB-1) has been implicated in the pathogenesis of inflammatory diseases. HMGB-1 has been proposed to be a crucial mediator in the pathogenesis of many diseases including sepsis, arthritis, cancer, autoimmunity diseases and diabetes. It has been suggested that HMGB-1 itself can signal through RAGEs (receptor for advanced glycation end products) and through the Toll-Like Receptors TLR2 and TLR4. Activation of these receptors results ultimately in the activation of Nuclear Factor-kappaB (NFkappaB), inducing the up-regulation of leukocyte adhesion molecules, production of pro-inflammatory cytokines and angiogenic factors in both hematopoietic and endothelial cells, thereby promoting inflammation. There are several patents proposed for controlling the production, secretion and neutralization of HMGB-1 and consequently the inflammatory process. We have divided the patents in six groups based on mechanism of action. The group 1 is associated with inhibition of HMGB-1 using anti-HMGB-1 antibodies; group 2: inhibition of HMGB-1 releases from the nucleus into the extracellular space; group 3: HMGB-A box as a competitive antagonist of HMGB-1; group 4: blockage of RAGE-HMGB-1 signaling using RAGE antagonists; group 5: blockage of TLR-HMGB-1 signaling using anti-TLR2 antibodies and group 6: other molecules that modulate HMGB-1 activity using e.g. human soluble thrombomodulin. The mechanism of HMGB-1 action, its role and efficiency of each group of patents proposed for controlling inflammation are discussed.


Surveillance for hepatocellular carcinoma in patients with advanced liver fibrosis.
Surveillance is the only pragmatic approach to improve treatment of hepatocellular carcinoma (HCC) owing to the fact that it allows detection of the tumor at an early and better curable stage. International liver societies recommend surveillance with biannual abdominal ultrasound (US) for patients with cirrhosis of any etiology because of their high risk of developing HCC. This strategy is considered cost-effective, as surveillance requires an articulated and costly set of interventions, including linkage to care of patients with an early detected tumor. However, as transition to HCC is increasingly being observed in noncirrhotic patients, the majority of which does not reach the threshold of cost effectiveness for screening. The European and Japanese liver societies elected to confine recommendations for HCC screening to noncirrhotic patients with advanced fibrosis due to hepatitis C or hepatitis B only. These latter recommendations, however, are challenged by the increasing number of patients with viral hepatitis in whom HCC risk has been attenuated but not eradicated by successful antiviral therapy. In this set of patients, entry criteria of surveillance need to be refined in the light of the suboptimal diagnostic accuracy of non invasive tests that are employed to identify the ideal candidates for surveillance.


Nuclear magnetic resonance studies of bone water.
Mineralized bone tissue has a significant water component. Bone water is associated with the collagen fibers or mineral fraction or occurring as pore water of the Haversian and lacunocanalicular system. Among the multiple physiologic functions that include signaling and providing to bone its viscoelastic properties, bone water enables the transport of ions and nutrients to and waste products from the cells. In addition, it plays a key role during mineralization whereby collagen-bound water is gradually replaced by calcium apatite-like mineral. In this review it is shown how nuclear magnetic resonance (NMR) allows the study of various physiologically relevant properties of bone water nondestructively. Isotope exchange experiments are described from which the apparent water diffusion coefficient can be calculated. The method is based on monitoring the migration of H2O into the D2O after immersion of the specimen in heavy water. Data obtained from rabbit cortical bone in the normal and mineral-depleted skeleton provide evidence for the hypothesized reciprocal relationship between bone water and mineral. Further, from the diffusion coefficient (Da = (7.8+/-1.5) x 10(-7) cm2/s) measured at 40 degrees C it can be inferred that diffusive transport of small molecules from the bone's microvascular system to the osteocytes occurs within minutes. Finally, whereas isotope exchange is not feasible in vivo, it is shown that bone water can be imaged by proton MRI.


Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells.
The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos. In order to clarify the function of HpEts in the gene regulatory network involved in PMC specification, we analyzed the zygotic expression pattern and the cis-regulatory region of HpEts, and examined the activity of the HpEts protein as a transcription factor. Intron-based PCR reveals that zygotic expression of HpEts starts at the cleavage stage, and that the rate of transcription reaches maximum at the unhatched blastula stage. A series of progressive deletions of the fragments from -4.2 kbp to +1206 bp of the HpEts, which directs PMC-specific expression, caused a gradual decrease in the specificity, implying that coordination of several cis-regulatory elements regulates the expression in PMCs. A minimum cis-element required for the temporal expression is located within a 10 bp from -243 bp to -234 bp. The HpEts protein remains in the cytoplasm of entire embryonic cells in the cleavage stage. At the unhatched blastula stage, the HpEts protein translocates into the nucleus in presumptive PMCs. Transactivation assays demonstrate that the HpEts protein activates a promoter of Spicule Matrix Protein 50 (SM50), which is a target of HpEts, which binds to the regulatory region of SM50.


Vibrational stark effect of the electric-field reporter 4-mercaptobenzonitrile as a tool for investigating electrostatics at electrode/SAM/solution interfaces.
4-mercaptobenzonitrile (MBN) in self-assembled monolayers (SAMs) on Au and Ag electrodes was studied by surface enhanced infrared absorption and Raman spectroscopy, to correlate the nitrile stretching frequency with the local electric field exploiting the vibrational Stark effect (VSE). Using MBN SAMs in different metal/SAM interfaces, we sorted out the main factors controlling the nitrile stretching frequency, which comprise, in addition to external electric fields, the metal-MBN bond, the surface potential, and hydrogen bond interactions. On the basis of the linear relationships between the nitrile stretching and the electrode potential, an electrostatic description of the interfacial potential distribution is presented that allows for determining the electric field strengths on the SAM surface, as well as the effective potential of zero-charge of the SAM-coated metal. Comparing this latter quantity with calculated values derived from literature data, we note a very good agreement for Au/MBN but distinct deviations for Ag/MBN which may reflect either the approximations and simplifications of the model or the uncertainty in reported structural parameters for Ag/MBN. The present electrostatic model consistently explains the electric field strengths for MBN SAMs on Ag and Au as well as for thiophenol and mercaptohexanoic acid SAMs with MBN incorporated as a VSE reporter.


Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea.
Gene transfer to a donor cornea ex vivo can modulate corneal graft failure in experimental animal models. We compared a lentiviral vector (LV) carrying the transgene ovine interleukin 10 (IL10) with a comparable adenoviral vector (Ad) for its ability to transduce ovine and human corneas and to modulate ovine corneal allograft survival. The LV carrying the ovine IL10 gene was used to transduce ovine and human corneas in vitro. LV-mediated gene expression in corneal endothelium was assessed by real-time quantitative reverse-transcriptase polymerase chain reaction, at varying doses and duration of transduction. The effect of ex vivo transduction of the donor cornea with LV-SV40-IL10 was assessed following orthotopic corneal transplantation in outbred sheep. Expression of IL10 mRNA in Ad-CMV-IL10-transduced ovine corneas was 10(3)-fold higher than in LV-SV40-IL10-transduced corneas (P < 0.0001), and 10(7)-fold higher than in non-transduced controls. IL10 was secreted rapidly from Ad-CMV-IL10-transduced, organ-cultured corneas, peaking at 13-15 days. IL10 secreted from LV-SV40-IL10-transduced corneas increased 20-fold compared with controls, but had not reached a plateau at 15 days. Gene expression driven by LV-SV40-IL10 varied with vector dose and transduction time, but was less than with Ad-CMV-IL10 at both mRNA and protein levels. Gene expression driven by LV-SV40-IL10 was faster in the human cornea than the ovine cornea. Corneal allograft survival was prolonged by a median of 7 days in the LV-SV40-IL10-treated recipients, compared with the control group (P = 0.026). Although lentiviral vectors show some promise for corneal gene therapy, they are less efficient than adenoviral vectors.


Research in bioanalysis and separations at the University of Nebraska - Lincoln.
The Chemistry Department at the University of Nebraska - Lincoln (UNL) is located in Hamilton Hall on the main campus of UNL in Lincoln, NE, USA. This department houses the primary graduate and research program in chemistry in the state of Nebraska. This program includes the traditional fields of analytical chemistry, biochemistry, inorganic chemistry, organic chemistry and physical chemistry. However, this program also contains a great deal of multidisciplinary research in fields that range from bioanalytical and biophysical chemistry to nanomaterials, energy research, catalysis and computational chemistry. Current research in bioanalytical and biophysical chemistry at UNL includes work with separation methods such as HPLC and CE, as well as with techniques such as MS and LC-MS, NMR spectroscopy, electrochemical biosensors, scanning probe microscopy and laser spectroscopy. This article will discuss several of these areas, with an emphasis being placed on research in bioanalytical separations, binding assays and related fields.


Toll-like receptors signaling contributes to immunopathogenesis of HBV infection.
Innate and adaptive immune systems have important role in the pathogenesis of acute and chronic infection with hepatitis B virus (HBV). These immune responses are mediated through complex interactions between the innate immune response and adaptive immune response. Toll-like receptors (TLRs) are a family of innate immune-recognition receptors that recognize the molecular patterns associated with microbial pathogens. So far, TLR1 to 13 were found in human or mice and investigated to detect the target molecules and the downstream mechanisms of these unique systems. Stimulation by their ligands initiates the activation of complex networks of intracellular signaling transduction and innate and adaptive immune-related cells (NK, NK-T, monocytes, dendritic cells, T cells, B cells, and Tregs, etc.). However, reports on such relationships between HBV and TLRs have been relatively rare in comparison to those on HCV and TLRs, but have recently been increasing. Thus, a review of TLRs involved in the pathogenesis of HBV infection may be needed toward better understanding of the immunopathogenesis of HBV infection.


Estimation of toothpaste fluoride intake in preschool children.
The objective of this study was to estimate the intake of toothpaste fluoride used by children aged 2 to 6 years (n=87) treated at a hospital of a medium-sized city (Campina Grande, PB) in the Northeastern region of Brazil. Data regarding sociodemographic characteristics of families and children's toothbrushing were collected from questionnaire-based interviews with parents/guardians, and the amount of fluoride used during toothbrushing was estimated using a precision scale for assessment of the risk of dental fluorosis, considering a cutoff value of 0.07 mgF/kg body weight/day. Fluoride content in the toothpastes was analyzed using a specific fluoride electrode. Data were analyzed using descriptive and inferential statistics using the chi-square and Fisher's exact tests (α=0.05). Considering the use of the derice, the risk of fluorosis in the children was 19.5%. There was significant association (p<0.05) between the risk of fluorosis, brushing frequency, type of derice and who performed the child's oral hygiene. It was concluded that a high percentage of children in the studied sample used toothpaste inappropriately and were at risk of developing dental fluorosis.


Herpes zoster after splenectomy. A study of patients without malignancy.
Follow-up information was obtained on 102 unselected patients who had undergone splenectomy for nonmalignant disease (ie, trauma, surgical complications, and hematologic indications). In 422 postsplenectomy years, three cases of herpes zoster were observed, two of which were associated with generalized cutaneous dissemination. This incidence is no greater than that reported elsewhere for age-matched normal population. We conclude that in patients without malignancy, splenectomy does not predispose to herpes zoster, but may play a role in cutaneous dissemination.


Inhibition by theophylline of the early component of canine ventricular contraction.
Changes in mechanical characteristics of the isolated canine ventricular muscle were investigated during interaction of isoproterenol with theophylline or caffeine. An early and a late component with time to peak tension of 80 and 150 ms, respectively, were differentiated in a single contraction of the muscle stimulated at 0.5 Hz at 37 degrees C during the interaction of isoproterenol and theophylline, or isoproterenol and caffeine. Isoproterenol increased preferentially the early component and affected only slightly the late one. Theophylline or caffeine elevated the early component less than the late one. In the presence of theophylline + isoproterenol or caffeine + isoproterenol the peak tension was achieved by a late component, whereas the increase in the early one induced by isoproterenol in 3 X 10(-7) M and higher was depressed significantly. During the interaction the rate of twitch relaxation was accelerated further rather than depressed. Changes in action potential indicate that the calcium influx via the myocardial cell membrane during depolarization was increased: the peak plateau potential was significantly elevated by theophylline alone and further by theophylline + isoproterenol. These results indicate that theophylline and caffeine (2 mM) may act intracellularly to inhibit the isoproterenol-induced promotion of the early component without impairing the isoproterenol-induced acceleration of relaxation in the canine ventricular muscle.


Dexamethasone and haemorrhage risk in paediatric tonsillectomy: a systematic review and meta-analysis.
Summary In children undergoing tonsillectomy, dexamethasone is recommended to reduce the risk of postoperative nausea and vomiting while non-steroidal anti-inflammatory drugs (NSAIDs) are used for pain relief. We aimed to determine whether children who receive dexamethasone or dexamethasone with NSAID are more likely to experience haemorrhage post-tonsillectomy. Randomized and non-randomized studies in which children undergoing tonsillectomy received dexamethasone or dexamethasone and NSAID were sought within bibliographic databases and selected tertiary sources. The risk of bias assessment and evaluation of haemorrhage rate data collection and reporting were assessed using the Cochrane Risk of Bias Tool and McHarm tool. Synthesis methods comprised pooled estimate of the effect of dexamethasone on the risk of haemorrhage rate using the Peto odds ratio (OR) method. The pooled estimate for haemorrhage rate in children who received dexamethasone was 6.2%, OR 1.41 (95% confidence interval 0.89-2.25, P=0.15). There was risk of bias and inconsistent data collection and reporting rates of haemorrhage in many of the included studies. Clinical heterogeneity was observed between studies. The pooled analysis did not demonstrate a statistically significant increase in the risk of post-tonsillectomy haemorrhage with dexamethasone with/without NSAID use in children. However, the majority of the included studies were not designed to investigate this endpoint, and thus large studies which are specifically designed to collect data on haemorrhage rate are needed.


Temperature dependence of the short-range repulsion between hydrated phospholipid membranes: A computer simulation study.
The temperature dependence of the short-range water-mediated repulsive pressure between supported phospholipid membranes is calculated at two intermembrane separations using the grand canonical Monte Carlo technique. At both separations, the simulated pressure tends to decrease with temperature, in qualitative agreement with the experimental measurements by Simon and co-workers [Simon et al., Biophys. J. 69, 1473 (1995)]. The decrease in pressure originates, at least in part, from a slight dehydration of the membranes and the associated reduction in the hydration component of the pressure.


Rh(III)-catalyzed selective mono- and dual-functionalization/cyclization of 1-aryl-5-aminopyrazoles with iodonium ylides.
An efficient Rh(III)-catalyzed selective mono- and dual-C-H bond functionalization/cyclization with iodonium ylide as a single coupling partner was demonstrated, in which fused benzodiazepine skeletons were obtained in excellent yields. This method greatly improved an effective approach to dual C-H unsymmetrical functionalization.


Prevalence of canaliculitis after smartplug insertion during long-term follow-up.
To evaluate the long-term prevalence of canaliculitis associated with the use of SmartPlugs. This retrospective study included 42 eyes of 25 dry eye patients at our institution who received a SmartPlug insertion between 2004 and 2008. Irrigation of the lacrimal system was performed in all patients prior to SmartPlug insertion to ensure the patency of the system. The median follow-up time was 7.9 (5.6-10.5) years after plug insertion. Six patients (seven eyes) of 25 patients (42 eyes) developed canaliculitis. The prevalence of canaliculitis per patient was 24%, and the prevalence of canaliculitis per SmartPlug was 16.6%. The median time from SmartPlug insertion to the onset of canaliculitis was 4.7 (1.4-6.0) years. The median age of the affected patients was 61.5 (36-74) years. Five of these patients were women and one was man. The pathogens were identified as Actinomyces in four eyes and as Staphylococcus aureus in three eyes. In five patients (six eyes), the canaliculitis was resolved by the application of topical antibiotics and repeated lacrimal irrigations with antibiotics and povidone-iodine. In one patient, a canaliculotomy was necessary to cure the condition. The late onset of complications after SmartPlug insertion requires long-term observation to identify the majority of complications associated with SmartPlugs. In our patients with a follow-up of up to 10.5 years, the use of SmartPlugs was associated with the highest prevalence of canaliculitis published thus far.


A single-stage posterior approach with open reduction and pedicle screw fixation in subaxial cervical facet dislocations.
The optimal treatment for cervical facet dislocations is controversial, but the generally accepted process recommends an initial closed reduction with the next step determined according to the success of the closed reduction and the presence of traumatic disc herniation. This study aimed to show the efficacy of a posterior approach performed with an open reduction and pedicle screw fixation with removal of disc particles, if required, in the management of subaxial cervical dislocations. Between March 2012 and September 2013, 21 consecutive patients with cervical facet dislocations were enrolled. The affected levels were as follows: 4 at C3-4; 2 at C4-5; 5 at C5-6; and 10 at the C6-7 level. Seven patients had traumatic disc herniations. Closed reduction was not attempted; a prompt posterior cervical surgery was performed instead. After open reduction, pedicle screw fixation was performed. In cases with traumatic disc herniation, herniated disc fragments were excised via a posterolateral approach and successful decompressions were determined by postoperative MRI studies. Clinical outcomes were assessed using the American Spinal Injury Association (ASIA) grading system. Radiological outcomes were assessed by comparing the degree of subluxation and the angle of segmental lordosis between pre- and postoperative CT scans. All patients improved neurologically. The mean segmental angles improved from 7.3° ± 8.68° to -5.9° ± 4.85°. The mean subluxation improved from 23.4% ± 16.52% to 2.6% ± 7.19%. Disc fragments were successfully removed from the 7 patients with herniated discs, as shown on MRI. Open reduction followed by pedicle screw fixation or posterolateral removal of herniated disc fragments is a good treatment option for cervical facet dislocations.


Immune gene profiles in Atlantic salmon (salmo salar L.) post-smolts infected with SAV3 by bath-challenge show a delayed response and lower levels of gene transcription compared to injected fish.
Salmonid alphavirus (SAV) causes pancreatic disease (PD) in salmonids in Northern Europe which results in large economic losses within the aquaculture industry. In order to better understand the underlying immune mechanisms during a SAV3 infection Atlantic salmon post-smolts were infected by either i.m.-injection or bath immersion and their immune responses compared. Analysis of viral loads showed that by 14 dpi i.m.-injected and bath immersion groups had 95.6% and 100% prevalence respectively and that both groups had developed the severe pathology typical of PD. The immune response was evaluated by using RT-qPCR to measure the transcription of innate immune genes involved in the interferon (IFN) response as well as genes associated with inflammation. Our results showed that IFNa transcription was only weakly upregulated, especially in the bath immersion group. Despite this, high levels of the IFN-stimulated genes (ISGs) such as Mx and viperin were observed. The immune response in the i.m.-injected group as measured by immune gene transcription was generally faster, and more pronounced than the response in the bath immersion group, especially at earlier time-points. The response in the bath immersion group started later as expected and appeared to last longer often exceeding the response in the i.m-injected fish at later time-points. High levels of transcription of many genes indicative of an active innate immune response were present in both groups.


The significance of atypical morphology in the changes of spectrum of postinfectious glomerulonephritis.
The characteristics of post infectious glomerulonephritis (PIGN) now differ from what were described decades ago. After encountering several patients of PIGN with atypical morphology, we conducted this retrospective study to determine the significance in the changes of clinicopathological spectrum of the disease. Between July 2000 and February 2009, 21 cases of PIGN were identified at a medical center in Taiwan. The patients' records were reviewed with respect to clinical presentation, microbiology, serology, morphology of renal biopsy, and clinical course. The mean age was 60.4 years. All patients developed acute renal failure and the majority (66.7%) required dialysis support. Hypocomplementemia was present in 61.9% of patients. The most frequently identified infectious agent was Staphylococcus (57.1%). Histological characteristics showed two distinct patterns of PIGN. One was diffuse endocapillary proliferation typical of PIGN (61.9%) and the other was atypical pattern of focal mesangial proliferation (38.1%). In comparison, glomerular neutrophil infiltration was more commonly present in typical pattern (p = 0.018). Glomerular IgA dominant or co-dominant deposition was more frequently seen in atypical pattern (p = 0.032). However, there were no statistically significant differences in the clinical presentation and outcome between the two groups. Our data also showed that the percentage of patients with atypical morphology PIGN significantly increased over time. Atypical pattern of focal mesangial proliferative glomerulonephritis may represent a resolution stage of PIGN. The nature of subclinical infection with a more protracted course may contribute to the increasing recognition of this resolving PIGN at the time of renal biopsy. Another possible explanation is that the atypical morphology may be a peculiar pattern of poststaphylococcal glomerulonephritis which was increasingly identified in PIGN over the past 10 years.


[Arterial blood supply to the laryngeal part of the pharynx].
The sources, anastomoses and variations of bloodsupply of the laryngeal part of the pharynx were studied in 100 corpses of different sex and age. It has been established that the fronto-lateral divisions of the laryngeal part of the pharynx are supplied with blood by pharyngeal branches of the superior and inferior paryngeal arteries. Ligation of the pharyngeal arteries during laryngectomy prior to their entering the larynx, i. e. before the divergence of the pharyngeal branches from them, as conventional, causes restriction of supply of these parts and can contribute to disjunction of the pharyngeal suture. The trunks of laryngeal arteries with their pharyngeal branches should be preserved, if possible. The posterior wall of the laryngeal part of the pharynx is divided into three zones depending on the main arterial sources (the ascending pharyngeal, superior and inferior thyroid arteries).


Detection and Characterization of Reactive Oxygen and Nitrogen Species in Biological Systems by Monitoring Species-Specific Products.
Since the discovery of the superoxide dismutase enzyme, the generation and fate of short-lived oxidizing, nitrosating, nitrating, and halogenating species in biological systems has been of great interest. Despite the significance of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in numerous diseases and intracellular signaling, the rigorous detection of ROS and RNS has remained a challenge. Recent Advances: Chemical characterization of the reactions of selected ROS and RNS with electron paramagnetic resonance (EPR) spin traps and fluorescent probes led to the establishment of species-specific products, which can be used for specific detection of several forms of ROS and RNS in cell-free systems and in cultured cells in vitro and in animals in vivo. Profiling oxidation products from the ROS and RNS probes provides a rigorous method for detection of those species in biological systems. Formation and detection of species-specific products from the probes enables accurate characterization of the oxidative environment in cells. Measurement of the total signal (fluorescence, chemiluminescence, etc.) intensity does not allow for identification of the ROS/RNS formed. It is critical to identify the products formed by using chromatographic or other rigorous techniques. Product analyses should be accompanied by monitoring of the intracellular probe level, another factor controlling the yield of the product(s) formed. More work is required to characterize the chemical reactivity of the ROS/RNS probes, and to develop new probes/detection approaches enabling real-time, selective monitoring of the specific products formed from the probes. Antioxid. Redox Signal. 28, 1416-1432.


Inhibition of Candida albicans by Fluvastatin Is Dependent on pH.
The cholesterol-lowering drug fluvastatin (FS) has an inhibitory effect on the growth of the pathogenic yeast Candida albicans that is dependent on the pH of the medium. At the low pH value of the vagina, FS is growth inhibitory at low and at high concentrations, while at intermediate concentrations (1-10 mM), it has no inhibitory effect. Examination of the effect of the common antifungal drug fluconazole in combination with FS demonstrates drug interactions in the low concentration range. Determination of intracellular stress and the activity of the FS target enzyme HMG-CoA reductase confirm our hypothesis that in the intermediate dose range adjustments to the sterol biosynthesis pathway can compensate for the action of FS. We conclude that the pH dependent uptake of FS across yeast membranes might make FS combination therapy an attractive possibility for treatment of vaginal C. albicans infections.


Effects of adhesive systems at different temperatures on the shear bond strength of orthodontic brackets.
Background. The temperature might affect the physical and mechanical properties of adhesive materials by reducing the polymerization rate. The present study aimed to evaluate the effect of temperature on the shear bond strength of metallic orthodontic brackets using various adhesive resin systems. Methods. Extracted human premolar teeth were randomly assigned to 8 groups (n=10) for bonding with the two available orthodontics adhesive systems (Transbond XT and NeoBond) at different temperatures: refrigeration temperature (4°C), room temperature (20°C), human body temperature (36°C) and high temperature (55°C). The shear bond strength (SBS) test was performed using a universal testing machine at a crosshead speed of 0.5 mm/min. The adhesive remnant index (ARI) was assigned to the fractured orthodontic brackets. Data were analyzed with one-way ANOVA, post hoc Tukey tests and independent t-test. Results. Transbond XT exhibited higher SBS values compared to Neobond at all the tested temperatures; however, a statistically significant difference was not observed (P>0.05). The SBS results were minimum at 4°C and maximum at 36°C in both the adhesive groups (P<0.05). Conclusion. Pre-heating orthodontic adhesives up to the body temperature prior to bonding the brackets in orthodontic treatment increased the bond strength of orthodontic brackets.


Validation of a Modified-Multidimensional Prognostic Index (m-MPI) including the Mini Nutritional Assessment Short-Form (MNA-SF) for the prediction of one-year mortality in hospitalized elderly patients.
The mortality prediction represents a key factor in the managing of elderly hospitalized patients. Since in older subjects mortality results from a combination of biological, functional, nutritional, psychological and environmental factors, a Multidimensional Prognostic Index (MPI) that predict short- and long-term mortality based on a standardized comprehensive geriatric assessment (CGA) has recently been developed and validated. This study compares the accuracy in predicting the mortality of the MPI with a modified version of the MPI (m-MPI) that included the Mini Nutritional Assessment-Short Form (MNA-SF) instead of the standard MNA. This prospective study with a one-year follow-up included 4088 hospitalized patients aged 65 years and older. A standardized CGA that included information on functional (Activities of Daily Living, ADL and Instrumental-ADL), cognitive (Short Portable Mental Status Questionnaire), risk of pressure sore (Exton-Smith Scale), comorbidities (CIRS Index), medications, living status and nutritional status (MNA and MNA-SF) was used to calculate the MPI using a previously validated algorithm. Higher MPI values were significantly associated with higher mortality rates with a close agreement between the estimated and the observed mortality both after 1-month (MPI1=2.8% versus m-MPI1=2.8%,p=0.946; MPI2=8.9% versus m-MPI2=9%,p=0.904; MPI3=21.9% versus m-MPI3=21.9,p=0.978) and 1-year of follow-up (MPI1=10.8% versus m-MPI1=10.5%,p=0.686; MPI2=27.3% versus m-MPI2=28%, p=0.495; MPI3=52.8% versus m-MPI3=52.7%,p=0.945). The estimated areas under the receiver operating characteristics (ROC) curves suggested a clinically negligible difference between the two indices. The m-MPI is as sensitive as the MPI in stratifying hospitalized elderly patients into groups at varying risk of short- and long-term mortality, but with fewer items.


Time-Course Evaluation of Body Mass Index in Japanese Children With Obstructive Sleep Apnea Syndrome After Adenotonsillectomy: A Three-Years Follow-Up Study.
Delayed physical growth is a common complication of pediatric obstructive sleep apnea syndrome (OSAS). Adenotonsillectomy (AT) is the first-line treatment for pediatric OSAS. Only a few studies have performed time-course BMI evaluation in pediatric OSAS patients post-operatively. Thus, we aimed to evaluate the time-course changes in pediatric OSAS patients after AT. Thirty-three children with OSAS who underwent AT were included and divided into two groups on the basis of their BMI z-scores (delayed physical growth group, n = 15; non-delayed physical growth group, n = 18). Clinical records of height and weight were collected before AT and at 6, 12, 24, and 36 months after AT. Changes in the mean BMI z-scores of the two groups were assessed up to 36 months. The mean BMI z-score was significantly increased in the delayed physical growth group at 6 months after AT. In contrast, the increase in mean BMI z-score was not observed in the non-delayed physical growth group. Growth improvement was noted in pediatric OSAS patients with delayed physical growth after AT. Our results suggest that AT is a promising therapy for improving the physical growth of pediatric OSAS patients with such problems.


[A comparative analysis of tonsillectomy and ablation of the palatine tonsils with CO2 laser].
Efficacy and toletance of two methods were compared: ablation of the palatine tonsils with CO2 laser and tonsillectomy. 270 case histories (118 after tonsillectomy and 152 after laser ablation of the palatine tonsils) have been analysed and 50 patients from each group have been examined. The comparison of the methods leads to conclusion that laser ablation of the palatine tonsils prevents intraoperative and postoperative complications; makes postoperative hemostatic therapy unnecessary; allows conduction of the operations in outpatient clinics or day hospitals; causes minimal pain in the postoperative period; shortens duration of disability.


The accuracy of inhomogeneity corrections in intensity modulated radiation therapy planning in Philips Pinnacle system.
The degree of accuracy of inhomogeneity corrections in a treatment planning system is dependent on the algorithm used by the system. The choice of field size, however, could have an effect on the calculation accuracy as well. There have been several evaluation studies on the accuracy of inhomogeneity corrections used by different algorithms. Most of these studies, however, focus on evaluating the dose in phantom using simplified geometry and open/static fields. This work focuses on evaluating the degree of dose accuracy in calculations involving intensity-modulated radiation therapy (IMRT) fields incident on a phantom containing both lung- and bone-equivalent heterogeneities using 6 and 10 MV beams. IMRT treatment plans were generated using the Philips Pinnacle treatment planning system and delivered to a phantom containing 55 thermoluminescent dosimeter (TLD) locations within the lung and bone and near the lung and bone interfaces with solid water. The TLD readings were compared with the dose predicted by the planning system. We find satisfactory agreement between planned and delivered doses, with an overall absolute average difference between measurement and calculation of 1.2% for the 6 MV and 3.1% for the 10 MV beam with larger variations observed near the interfaces and in areas of high-dose gradient. The results presented here demonstrate that the convolution algorithm used in the Pinnacle treatment planning system produces accurate results in IMRT plans calculated and delivered to inhomogeneous media, even in regions that potentially lack electronic equilibrium.


Meningococcal disease: shifting epidemiology and genetic mechanisms that may contribute to serogroup C virulence.
During the past decade, monovalent serogroup C and quadrivalent (serogroups A, C, W135, Y) meningococcal vaccination programs have been introduced in multiple industrialized countries. Many of these programs have been successful in reducing the burden of disease due to vaccine-preventable serogroups of Neisseria meningitidis in target age groups. As a result, disease burden in these countries has decreased and is primarily serogroup B, which is not vaccine preventable. Despite the success of these programs, meningococcal disease continues to occur and there is always concern that serogroup C organisms will adapt their virulence mechanisms to escape pressure from vaccination. This review highlights the current epidemiology of meningococcal disease in Europe and United States, as well as genetic mechanisms that may affect virulence of serogroup C strains and effectiveness of new vaccines.


Detection of in vivo DNA repair synthesis in mouse liver and lung induced by treatment with benzo(a)pyrene or 4-nitroquinoline 1-oxide.
We examined in vivo DNA repair synthesis in liver and lung of A/HeJ mice treated with benzo(a)pyrene (BP) or 4-nitroquinoline 1-oxide. To differentiate between the removal of carcinogen metabolite:DNA adducts due to cell turnover and DNA repair, we measured unscheduled DNA synthesis (UDS) in the nonreplicating DNA fraction. Mice were exposed to bromodeoxyuridine pellets 1 hr prior to carcinogen treatment. Immediately following carcinogen exposure, mice received 4 hourly i.v. doses of [3H]thymidine. Mice were sacrificed 5 hr post-carcinogen treatment, and DNA was isolated. Purified DNA was then separated into newly replicated and nonreplicated DNA by ultracentrifugation in alkaline CsCl gradients. BP induced UDS in the liver at p.o. doses of 0.3 and 3.0 mg/mouse, whereas we failed to detect UDS in the lung. However, 4-nitroquinoline 1-oxide, another lung carcinogen, induced a definite repair response in the lung but not in the liver. It is not clear why mouse lung cells have the capacity to repair 4-nitroquinoline 1-oxide-induced damage to DNA and not the damage induced by BP, since both of these lung carcinogens form bulky adducts with DNA. These results demonstrate that (a) the in vivo disappearance of BP metabolite:DNA adducts from the lung of the A/HeJ mouse is due to cell turnover, whereas the disappearance of adducts from the liver is due, in part, to DNA repair and (b) induction of in vivo UDS after treatment with two different lung carcinogens is both tissue and carcinogen dependent in this mouse strain.


In vitro cytotoxicology model of oligo-chitosan and n, o-carboxymethyl chitosan using primary normal human epidermal keratinocyte cultures.
Chitosan (beta-1, 4-D-glucosamine) is a deacetylated form of chitin with excellent biological properties in wound management. The natural properties of chitosan have the physical and chemical limitations to be widely used in biomedical fields. The improvement of the physical and chemical properties of chitosan with some additional chemicals will alter its biocompatibility. Therefore, the biological attribute of the modified chitosan must be evaluated. In this study, the cytotoxicity of oligo-chitosan (OC) and N, O- carboxymethyl-chitosan (NO-CMC) derivatives (O-C 1%, O-C 5%, NO-CMC 1% and NO-CMC 5%) was evaluated using primary normal human epidermal keratinocyte (pNHEK) cultures as an in vitro toxicology model at standardized cell passages (fourth passages). 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl tetrazolium bromide (MTT) was used as a cell viability assay. The O-C 1% is one of the most compatible chitosan derivatives because it steadily sustained >70% of viable cells until 72 hr post-treatment. This was followed by O-C 5%, NO-CMC 5% and NO-CMC 1%. Therefore, oligo-chitosan had the ideal properties of a biocompatible material compared to N, O- carboxymethyl-chitosan in this study.


Development of hospital information systems: user participation and factors affecting it.
Given the large volume of data generated in hospitals, in order to efficiently management them; using hospital information system (HIS) is critical. User participation is one of the major factors in the success of HIS that in turn leads Information needs and processes to be correctly predicted and also their commitment to the development of HIS to be augmented. The purpose of this study is to investigate the participation rate of users in different stages of HIS development as well as to identify the factors affecting it. This is a descriptive-cross sectional study which was inducted in 2014. The study population consists of 140 HIS users (from different types of job including physicians, nurses, laboratory, radiology and HIM staffs) from Teaching Hospitals Affiliated to Urmia University of Medical Sciences. Data were collected using a self-structured questionnaire which was estimated as both reliable and valid. The data were analyzed by SPSS software descriptive statistics and analytical statistics (t-test and chi-square). The highest participation rate of users in the four-stage development of the HIS was related to the implementation phase (2.88) and the lowest participation rate was related to analysis (1.23). The test results showed that the rate of user participation was not satisfactory in none of the stages of development (P< 0.05). The most important factors in increasing user participation include established teamwork from end-users and the support of top managers from HIS development. According to the results obtained from the study, it seems that health care administrators must have a detailed plan for user participation prior to the development and purchase of HIS so that they identify the real needs as well as increase their commitment and motivations to develop, maintain and upgrade the system, and in this way, the success of the system will be assured.


Antenatal Bartter's syndrome: why is this not a lethal condition?
There are four themes in this teaching exercise for Professor McCance. The first challenge was to explain how a premature infant with Bartter's syndrome could survive despite having such a severe degree of renal salt wasting. Second, the medical team wanted to know why there was such a dramatic decrease in the natriuresis in response to therapy, despite the presence of a permanent molecular defect that affected the loop of Henle. Third, Professor McCance was asked why this patient seemed to have a second rare disease, AQP2 deficiency type of nephrogenic diabetes insipidus. The fourth challenge was to develop a diagnostic test to help the parents of this baby titrate the dose of indomethacin to ensure an effective dose while minimizing the likelihood of developing nephrotoxicity. The missing links in this interesting story emerge during a discussion between the medical team and its mentor.


[A consideration through the support to resident diabetic patient and care giver].
Our hospital is a National General Hospital with 585 beds. We began the visiting care service from 1990 and four visiting staffs are working at present. The number of targets was 69 in 2002 including 32 patients over 70 years old and 20 care-givers over 70 years old. Visiting care has been conduced to a 72-year-old female with diabetes since July 2000. The patient is in bedridden condition and at the beginning of visiting, she was taking oral medication but the condition was worsen by poor glycemic control and changed to insulin injection from June 2002 after admission to the hospital repeatedly. The patient undergoes the measurement of blood sugar daily and takes meals with 1,200 to 1,400 kcal prepared by her husband. The care-giver is a 71 year-old husband. He was an "all-work, no-play" type of person and had never done the housework, but he started to manage both housework and nursing because of bedridden of his wife. He is a reticent theorist, hates illogical behavior, and does not swayed by other opinions. He has accepted the things which need new knowledge and techniques such as measurement of blood glucose and insulin injection. However, for meals, he only bought side dishes and placed them. Care such as keeping the patient clean was in a same state. The patient consulted and admission to the hospital repeatedly because conditions were not stable. Visiting nurse supported daily life of patient and care giver especially in nutrition instruction to continue home life. As the result, attitude and behavior toward nursing of care-giver were changed and the patient could continue home life. Therefore we reported here.


Tea drinking as a risk factor for neural tube defects in northern China.
Tea catechins inhibit the activity of the enzyme dihydrofolate reductase, which catalyzes the inactive form of folate 7,8-dihydrofolate to the active form of folate 5,6,7,8-tetrahydrofolate. This inhibition leads to disturbance of the folate metabolic pathway and to lower bioavailability of folate to cells; therefore, it may increase the risk of neural tube defects (NTDs) in a developing embryo. We examined the association between tea drinking during the periconceptional period and the risk of NTDs. Cases were fetuses or neonates with an NTD as ascertained through a population-based surveillance system in 4 rural counties of Shanxi Province in northern China during 2002 through 2007. Controls were term infants without congenital malformations. Demographic, reproductive, and exposure data were obtained through a face-to-face interview. A total of 631 NTD cases and 857 controls were included in the analysis. Compared with women who did not drink tea during the periconceptional period (487 cases and 688 controls), women who drank tea daily (20 cases and 9 controls) had a 3-fold increased risk (odds ratio = 3.1 [95% confidence interval = 1.4-7.0]) of having an NTD-affected pregnancy. The elevated risk associated with daily tea drinking remained after adjusting for maternal age, educational level, occupation, and periconceptional folic acid supplementation (3.4 [1.4-8.3]). The association was present for all 3 major subtypes of NTDs (ie, anencephaly, spina bifida, and encephalocele). Daily tea drinking during the periconceptional period was associated with an elevated risk of NTDs in this Chinese population.


Susceptibility and diversity in the therapy-refractory genus scedosporium.
Scedosporium species show decreased susceptibility to the majority of systemic antifungal drugs. Acquired resistance is likely to disseminate differentially with the mode of exchange of genetic material between lineages. Inter- and intraspecific diversities of Scedosporium species were analyzed for three partitions (rDNA internal transcribed spacer gene [ITS], partial β-tubulin gene, and amplified fragment length polymorphism profiles), with the aim to establish distribution of resistance between species, populations, and strains. Heterogeneity of and recombination between lineages were determined, and distances between clusters were calculated using a centroid approach. Clinical, geographic, and antifungal data were plotted on diversity networks. Scedosporium minutisporum, Scedosporium desertorum, and Scedosporium aurantiacum were distinguished unambiguously in all partitions and had differential antifungal susceptibility profiles (ASP). Pseudallescheria fusoidea and Pseudallescheria ellipsoidea were indistinguishable from Scedosporium boydii. Pseudallescheria angusta took an intermediate position between Scedosporium apiospermum and S. boydii. Scedosporium boydii and S. apiospermum had identical ASP. Differences in (multi)resistance were linked to individual strains. S. apiospermum and S. boydii showed limited interbreeding and were recognized as valid, sympatric species. The S. apiospermum/S. boydii group, comprising the main clinically relevant Scedosporium species, consists of separate lineages and is interpreted as a complex undergoing sympatric evolution with incomplete lineage sorting. In routine diagnostics, the lineages in S. apiospermum/S. boydii are indicated with the umbrella descriptor "S. apiospermum complex"; individual species can be identified with rDNA ITS with 96.3% confidence. Voriconazole is recommended as the first-line treatment; resistance against this compound is rare.


The less-often-traveled surface of stem cells: caveolin-1 and caveolae in stem cells, tissue repair and regeneration.
Stem cells are an important resource for tissue repair and regeneration. While a great deal of attention has focused on derivation and molecular regulation of stem cells, relatively little research has focused on how the subcellular structure and composition of the cell membrane influences stem cell activities such as proliferation, differentiation and homing. Caveolae are specialized membrane lipid rafts coated with caveolin scaffolding proteins, which can regulate cholesterol transport and the activity of cell signaling receptors and their downstream effectors. Caveolin-1 is involved in the regulation of many cellular processes, including growth, control of mitochondrial antioxidant levels, migration and senescence. These activities are of relevance to stem cell biology, and in this review evidence for caveolin-1 involvement in stem cell biology is summarized. Altered stem and progenitor cell populations in caveolin-1 null mice suggest that caveolin-1 can regulate stem cell proliferation, and in vitro studies with isolated stem cells suggest that caveolin-1 regulates stem cell differentiation. The available evidence leads us to hypothesize that caveolin-1 expression may stabilize the differentiated and undifferentiated stem cell phenotype, and transient downregulation of caveolin-1 expression may be required for transition between the two. Such regulation would probably be critical in regenerative applications of adult stem cells and during tissue regeneration. We also review here the temporal changes in caveolin-1 expression reported during tissue repair. Delayed muscle regeneration in transgenic mice overexpressing caveolin-1 as well as compromised cardiac, brain and liver tissue repair and delayed wound healing in caveolin-1 null mice suggest that caveolin-1 plays an important role in tissue repair, but that this role may be negative or positive depending on the tissue type and the nature of the repair process. Finally, we also discuss how caveolin-1 quiescence-inducing activities and effects on mitochondrial antioxidant levels may influence stem cell aging.


Risk of group A meningococcal disease: bacterial interference and cross-reactive bacteria among mucosal flora.
During outbreaks of group A meningococcal disease in Seattle, Wash., and Portland, Oreg., we studied the mucosal flora of the affected population and comparison groups to identify possible determinants of susceptibility and resistance to disease. Antimeningococcal immunoglobulin A can block the bactericidal activity of specific antibodies of other classes and has been associated with susceptibility in adults. We used immunoprecipitation and fluorescent-antibody techniques to detect mucosal microorganisms cross-reactive with group A meningococci that might have stimulated such antibodies. Cross-reactive strains of Bacillus pumilus and Streptococcus faecalis were found. Bacterial interference on mucosal surfaces has been shown to reduce susceptibility to other pathogens. With an agar overlay technique, we sought nasopharyngeal microorganisms that inhibited the growth of group A meningococci. Forty-five percent of subjects carried inhibitory strains representing at least nine different species. Inhibitory strains were less common (32%) in residents from "skid row" areas (see D.J. Bogue, Skid Row in American Cities, University of Chicago Press, for a comprehensive definition of these areas) than in a comparison group that did not experience meningococcal disease (61%), suggesting that their presence may be associated with resistance to acquisition of meningococci or to meningococcal disease.


In-situ synthesis of Ni-Co-S nanoparticles embedded in novel carbon bowknots and flowers with pseudocapacitance-boosted lithium ion storage.
We design a facile approach to prepare a bimetallic transition-metal-sulphide-based 3D hierarchically-ordered porous electrode based on bimetallic metal-organic frameworks (Ni-Co-MOFs) by using confinement growth and in-situ sulphurisation techniques. In the novel resulting architectures, Ni-Co-S nanoparticles are confined in bowknot-like and flower-like carbon networks and are mechanically isolated but electronically well-connected, where the carbon networks with a honeycomb-like feature facilitate electron transfer with uninterrupted conductive channels from all sides. Moreover, these hierarchically-ordered porous structures together with internal voids can accommodate the volume expansion of the embedded Ni-Co-S nanoparticles. The pseudocapacitive behaviours displayed in the NCS@CBs and NCS@CFs occupied a significant portion in the redox processes. Because of these merits, both the as-built bowknot and flower networks show excellent electrochemical properties for lithium storage with superior rate capability and robust cycling stability (994 mAh g-1 for NCS@CBs and 888 mAh g-1 for NCS@CFs after 200 cycles). This unique 3D hierarchically-ordered structural design is believed to hold great potential applications in propagable preparation of carbon networks teamed up with sulphide nanocrystals for high energy storage.


Rapid analysis of trace volatile formaldehyde in aquatic products by derivatization reaction-based surface enhanced Raman spectroscopy.
Toxic formaldehyde is sometimes used illegally as a food preservative, however, on-site rapid analysis of trace formaldehyde in aquatic products remains a challenge. In this work, a simple on-site rapid quantification method for trace volatile formaldehyde in aquatic products was developed by a derivative reaction-based surface enhanced Raman spectroscopy (SERS) technique coupled with a homemade portable purge-sampling device. Trace formaldehyde separated from complicated aquatic matrices via a purge-sampling procedure was reacted with a derivative reagent to produce a Raman-active analyte for consequent SERS analysis. Au/SiO2 nanoparticles (NPs) were employed as the enhancement substrate to achieve significant enhancement of Raman signal intensity. Conditions of derivative reaction and SERS detection were optimized in detail, and the selectivity of this analytical method was also evaluated based on related analogs. Under optimal conditions, an extremely low detection limit of 0.17 μg L(-1) was achieved. Trace volatile formaldehyde can be found in fresh squid and shrimp samples without obvious matrix interference, and this was quantified to be 0.13-0.21 mg kg(-1) using the described method. The recoveries of spiked aquatic product samples were found to be 70.0-89.1% with RSDs of 2.3-7.2% (n = 3). The results suggest that the proposed method is reliable and suitable for on-site rapid analysis of trace formaldehyde in aquatic products.


[Mapping of genes 14 and 16 of phage phi 80].
This study was performed to obtain more detailed information on the early region of phage phi 80. For this purpose, a selective method for isolation of early phage ts mutants was developed. 32 ts mutants of phi 80 obtained and early sus mutants isolated by Sato were characterized by complementation tests and deletion mapping. The results of this study differ from those of Sato in two aspects. Firstly, it was shown that sus mutations 250, 258 and 326 of phi 80 define one gene 16, and that gene 15 previously determined by Sato via sus 326 mutation does not exist. Secondly, we found that the order of the phi 80 early genes 14 and 16 is cI-16-14, contrary to the Sato data (cI-14-16).


Older doctors and progression through specialty training in the UK: a cohort analysis of General Medical Council data.
To determine whether older age at graduation is associated with any difference in outcomes from the annual specialty training progression assessment. An open cohort of 38 308 doctors who graduated from a UK medical school with annual assessments of progression in their specialty training programme with data centrally collected by the General Medical Council between 5 August 2009 to 31 July 2012. Mature junior doctors (≥29 years at graduation) were more likely to have problems with progression on their annual review of competence progression record of in training assessment (ARCP/RITA) than their younger colleagues (OR 1.34, 95% CI 1.22 to 1.49, p<0.001). This association was, if anything, even stronger (OR 1.57, 95% CI 1.41 to 1.74, p<0.001) after adjustment for gender, ethnicity, type of University and specialty. The same was true when only looking at the most extreme ARCP outcome (4) which is being asked to leave their specialist programme (OR 1.81, 95% CI 1.34 to 2.44, p<0.001). Mature doctors are a growing part of the medical workforce and they are likely to broaden the spectrum of doctors by bring different life experience to the profession. These results suggest that they are more likely to have problems with progressing through their specialist training programme. More research is required to determine the reasons behind these associations and how mature doctors can be supported both in choosing the best training programme and in coping with the complex demands of higher training at a later stage in their lives.


The functions of autobiographical memory: an integrative approach.
Recent research in cognitive psychology has emphasised the uses, or functions, of autobiographical memory. Theoretical and empirical approaches have focused on a three-function model: autobiographical memory serves self, directive, and social functions. In the reminiscence literature other taxonomies and additional functions have been postulated. We examined the relationships between functions proposed by these literatures, in order to broaden conceptualisations and make links between research traditions. In Study 1 we combined two measures of individual differences in the uses of autobiographical memory. Our results suggested four classes of memory functions, which we labelled Reflective, Generative, Ruminative, and Social. In Study 2 we tested relationships between our four functions and broader individual differences, and found conceptually consistent relationships. In Study 3 we found that memories cued by Generative and Social functions were more emotionally positive than were memories cued by Reflective and Ruminative functions. In Study 4 we found that reported use of Generative functions increased across the lifespan, while reported use of the other three functions decreased. Overall our findings suggest a broader view of autobiographical memory functions that links them to ways in which people make meaning of their selves, their environment, and their social world more generally.


[Ambulatory sclerosurgical treatment of varicose disease of the lower extremities].
The results of ambulatory sclerosurgical treatment of 258 patients, suffering varicose disease of the lower extremities (VDLE), were analyzed. The complications revealed are not severe and do not worsen the treatment results in whole. Application of intraoperative microfoam scleroobliteration of the stem veins have had guaranteed their complete occlusion in 97.5% of the patients. Sclerosurgical treatment have promoted a trustworthy (P < 0.0001) reduction of rate of chronic venous insufficiency (CHVI) class C2-C5 (in accordance with CEAP classification), and estimation of the CHVI severity classes C0, C1 witnesses the efficacy of this method of the VDLE treatment.


Effect of caffeine on induction of endogenous type C virus in mouse cells in vitro.
The effect of caffeine on the expression of murine endogenous virus in mouse cells induced by radiation and chemicals was studied. Postirradiation treatment of K-BALB cells with caffeine enhanced cell killing as well as the induction of xenotropic virus after ultraviolet light irradiation. The degree of enhancement for the virus induction was comparable to that for cell killing. On the other hand, colony-forming ability and the expression of xenotropic virus of K-BALB cells after X-irradiation were unaffected by caffeine. These data suggest a linear relationship between the degree of endogenous virus expression and the amount of lethal damages after irradiation. For induction by halogenated pyrimidines, a 24-hr incubation of AKR2B cells with caffeine after 5-iodo-2'-deoxyuridine treatment resulted in marked suppression of the expression of ecotropic virus. On the contrary, in K-BALB cells, caffeine exerted only a small effect on 5-iodo-2'-deoxyuridine-induced expression of ecotropic and xenotropic viruses. These results indicate that, although using the same inducing agent, the pathway of endogenous virus induction may be different for AKR2B cells and for K-BALB cells.


Reasons that orthodontic faculty teach and consider leaving teaching.
To assess reasons why orthodontic faculty consider leaving academia, a pretested questionnaire was mailed to 200 full-time (FT) and 200 part-time (PT) faculty randomly selected from the United States and Canada. A total of 144 (72 percent) of FT and 120 (60 percent) PT responded. About 38 percent FT and 25 percent PT were currently considering leaving academia. The average age of each group was about fifty years. Although significant differences were found in fifteen factors affecting the decision to leave, three factors ranked as most important (means> or =3.4) for FTs: salary support, financial support of department, and control over work or destiny. The three factors most important (means>3. 1) for PTs were: challenge of private practice, family commitments, and personal. FT and PT were similar in the most important and least important factors influencing their initial reasons to teach and satisfaction in teaching. However, the reasons why FT considered leaving were work related, while the PT's reasons were more personal. With the current shortage of FT orthodontic faculty becoming imminently greater, it appears that work-related issues could be addressed directly by administration.


Angiotensin antagonists as diagnostic and pharmacologic tools.
Angiotensin antagonists have become useful tools in studying pharmacologic and pathophysiologic roles of the renin-angiotensin axis. Several of these uses are described herein. Their value as t-ols in the diagnosis of renal artery stenosis is yet to be determined.


Pain as Practice in Paolo Mantegazza's Science of Emotions.
Paolo Mantegazza's science of emotions represents the dominant style of thinking that was fostered by the late nineteenth-century Italian scientific community, a positivist school that believed that the dissemination of Darwin's evolutionary ideas would promote social progress in that country. Within this collective thought, Mantegazza was committed not only to studying the physiological experience of pain by means of vivisection but also to completing an anthropological study that examined the differences between the expressions of suffering in primitive and civilized cultures.Thus, the meaning of pain appears throughout Mantegazza's research as a result of applying an ensemble of scientific practices integral to observation, experimentation, and he scientific self, which enabled its main physiological and psychological manifestations to be reproduced in the laboratory. Among these practices, photography allowed Mantegazza to mobilize pain as an emotion whose performativity shaped national identities, such as those that embodied the recently created Italian state.


Enhanced Recovery After Surgery strategies for elective craniotomy: a systematic review.
Enhanced Recovery After Surgery (ERAS) has led to a paradigm shift in perioperative care through multimodal interventions. Still, ERAS remains a relatively new concept in neurosurgery, and there is no summary of evidence on ERAS applications in cranial neurosurgery. The authors systematically reviewed the literature using the PubMed/MEDLINE, Embase, Scopus, and Cochrane Library databases for ERAS protocols and elements. Studies had to assess at least one pre-, peri-, or postoperative ERAS element and evaluate at least one of the following outcomes: 1) length of hospital stay, 2) length of ICU stay, 3) postoperative pain, 4) direct and indirect healthcare cost, 5) complication rate, 6) readmission rate, or 7) patient satisfaction. A final 27 articles were included in the qualitative analysis, with mixed quality of evidence ranging from high in 3 cases to very low in 1 case. Seventeen studies reported a complete ERAS protocol. Preoperative ERAS elements include patient selection through multidisciplinary team discussion, patient counseling and education to adjust expectations of the postoperative period, and mental state assessment; antimicrobial, steroidal, and antiepileptic prophylaxes; nutritional assessment, as well as preoperative oral carbohydrate loading; and postoperative nausea and vomiting (PONV) prophylaxis. Anesthesiology interventions included local anesthesia for pin sites, regional field block or scalp block, avoidance or minimization of the duration of invasive monitoring, and limitation of intraoperative mannitol. Other intraoperative elements include absorbable skin sutures and avoidance of wound drains. Postoperatively, the authors identified early extubation, observation in a step-down unit instead of routine ICU admission, early mobilization, early fluid de-escalation, early intake of solid food and liquids, early removal of invasive monitoring, professional nutritional assessment, PONV management, nonopioid rescue analgesia, and early postoperative imaging. Other postoperative interventions included discharge criteria standardization and home visits or progress monitoring by a nurse. A wide range of evidence-based interventions are available to improve recovery after elective craniotomy, although there are few published ERAS protocols. Patient-centered optimization of neurosurgical care spanning the pre-, intra-, and postoperative periods is feasible and has already provided positive results in terms of improved outcomes such as postoperative pain, patient satisfaction, reduced length of stay, and cost reduction with an excellent safety profile. Although fast-track recovery protocols and ERAS studies are gaining momentum for elective craniotomy, prospective trials are needed to provide stronger evidence.


Water, Sanitation, and Hygiene Practices Associated with Nutritional Status of Under-Five Children in Semi-Pastoral Communities Tanzania.
Undernutrition among under-five children is a public health concern in developing countries and has been linked with poor water, sanitation, and hygiene (WASH) practices. This study aimed at assessing WASH practices and its association with nutritional status of under-five children in semi-pastoral communities of Arusha. The study was cross-sectional in design. Mother-child pairs from 310 households in four villages of Monduli and Longido were involved. Weight and height of children were measured using weighing scale and length/height board, respectively. Children's age was recorded using clinic cards. Hemoglobin level of each child was tested using Hemo Cue® Hb 201 + photometer (HemoCue AB, Ängelholm, Sweden) machine. Structured questionnaire was used to gather information on WASH, child morbidity, demographic, and sociocultural characteristics. Prevalence of stunted, underweight, wasted, anemia, and diarrhea were 31.6%, 15.5%, 4.5% 61.2%, and 15.5%, respectively. Children with diarrhea 2 weeks preceding the survey (P = 0.004), children using surface water for domestic purposes (P < 0.001), and those with uneducated mothers (P = 0.001) had increased risk of being stunted and underweight. Children introduced to complementary foods before 6 months of age (P = 0.02) or belonging to polygamous families (P = 0.03) had increased risk of being stunted. Consumption of cow's milk that is not boiled (P = 0.05) or being a boy (P = 0.03) was associated with underweight. Prevalence of undernutrition among under-five children in the population under study was alarming and it could be associated with poor WASH practices and other sociocultural factors. This study underlines the importance of incorporating WASH strategies in formulation of interventions targeting on promotion of nutrition and disease prevention in pastoral communities.


Blunt chest trauma: is there a place for rib stabilization?
Rib fractures are a common and highly morbid finding in patients with blunt chest trauma. Over the past decade, a renewed interest in (and instrumentation for) rib fixation in this cohort has occurred. Stabilization of the chest wall in this setting, particularly when a flail segment is present, is associated with significant reductions in the rates of respiratory failure, pneumonia, ICU stay, and mortality. Thoracic surgeons should remain actively involved in this evolving area of our specialty to further optimize patient outcomes.


Uptake of a colorectal cancer screening blood test in people with elevated risk for cancer who cannot or will not complete a faecal occult blood test.
Participation rates in colorectal cancer (CRC) screening programmes using faecal occult blood tests (FOBTs) are low. Nonparticipation is commonly attributed to psychosocial factors, but some medical conditions also prevent screening. These barriers might be partially overcome if a blood test for CRC screening was available. This study determined whether people who had always declined screening by FOBT would participate if offered a blood test. An audit of registrants within a personalized CRC screening programme was undertaken to determine the reasons for regular nonparticipation in FOBT. Consistent nonparticipants (n=240) were randomly selected and invited for CRC screening with a blood test. Demographic characteristics and the reasons for prior FOBT nonparticipation were collected by means of a questionnaire. Nonparticipation in the screening programme could be classified as either behavioural (8.6%), with consistent noncompliance, or due to medical contraindications (8.5%), which included chronic rectal bleeding, being deemed unsuitable by a health professional, and needing personal assistance. Blood test uptake was 25%, with participation in the medical contraindications group greater than that in the behavioural group (43 vs. 12%, P<0.001). Reported behavioural reasons for nonparticipation in faecal immunochemical test included procrastination and dislike of the test, but these were not associated with blood test uptake (P>0.05). There is a subgroup of the community who have medical reasons for nonparticipation in CRC screening with FOBT but will participate if offered a blood test. The option of a blood test does not, however, improve uptake in those who admit to behavioural reasons for noncompliance with screening.


Angioplasty alone versus angioplasty and stenting for subclavian artery stenosis--a systematic review and meta-analysis.
Subclavian artery stenosis has long been treated with great success with bypass surgery. Percutaneous intervention, often used in combination with stent placement, has come into vogue for the past few years as a safe and effective therapeutic modality. This study aimed to compare angioplasty alone with angioplasty followed by stent placement by combining available data. The objective of this study was to perform a review of the available literature to compare the efficacy of percutaneous transluminal angioplasty (PTA) alone with PTA followed by stent placement for proximal subclavian artery stenosis. Successful recanalization was defined as patency at the end of 1 year, and reocclusions and restenoses were noted as events for the purpose of pooling the data. The authors searched the Specialized Register and the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, PubMed, EMBASE, and CINAHL databases for relevant trials/studies comparing PTA and PTA with stenting. Review authors independently assessed the methodological quality of studies (focusing on the adequacy of the randomization process, allocation concealment, blinding, completeness of follow-up, and intention-to-treat analysis) and selected studies for inclusion. All retrospective observational studies were also included in the analysis in the absence of double-blinded randomized trials for increasing sample size. All analyses were done using RevMan 5.0. Odds ratio was calculated using Mantel-Haenszel test with a fixed effect model. All included studies were assessed by all authors for potential sources of bias. Eight studies were included in the analysis having 544 participants. Stenting after PTA was significantly superior to angioplasty alone for treatment of subclavian artery stenosis and maintenance of patency at 1 year, as indicated by absence of events (P = 0.004; 95% confidence interval, odds ratio 2.37 [1.32-4.26]) without significant complication rates for either procedure. There is evidence in favor of stent placement after angioplasty for successful recanalization of stenosed subclavian arteries and long-term maintenance of patency without significant increase in risk for major complications in subjects.


PlantID - DNA-based identification of multiple medicinal plants in complex mixtures.
An efficient method for the identification of medicinal plant products is now a priority as the global demand increases. This study aims to develop a DNA-based method for the identification and authentication of plant species that can be implemented in the industry to aid compliance with regulations, based upon the economically important Hypericum perforatum L. (St John's Wort or Guan ye Lian Qiao). The ITS regions of several Hypericum species were analysed to identify the most divergent regions and PCR primers were designed to anneal specifically to these regions in the different Hypericum species. Candidate primers were selected such that the amplicon produced by each species-specific reaction differed in size. The use of fluorescently labelled primers enabled these products to be resolved by capillary electrophoresis. Four closely related Hypericum species were detected simultaneously and independently in one reaction. Each species could be identified individually and in any combination. The introduction of three more closely related species to the test had no effect on the results. Highly processed commercial plant material was identified, despite the potential complications of DNA degradation in such samples. This technique can detect the presence of an expected plant material and adulterant materials in one reaction. The method could be simply applied to other medicinal plants and their problem adulterants.


Endoplasmic Reticulum Homeostasis Regulates TLR4 Expression and Signaling in Mast Cells.
The endoplasmic reticulum (ER) is a dynamic organelle that responds to demand in secretory proteins by undergoing expansion. The mechanisms that control the homeostasis of ER size and function involve the activation of the unfolded protein response (UPR). The UPR plays a role in various effector functions of immune cells. Mast cells (MCs) are highly granular tissue-resident cells and key drivers of allergic inflammation. Their diverse secretory functions in response to activation through the high-affinity receptor for IgE (FcεRI) suggest a role for the UPR in their function. Using human cord blood-derived MCs, we found that FcεRI triggering elevated the expression level and induced activation of the UPR transducers IRE1α and PERK, accompanied by expansion of the ER. In mouse bone marrow-derived MCs and peritoneal MCs, the ER underwent a more moderate expansion, and the UPR was not induced following MC activation. The deletion of IRE1α in mouse MCs did not affect proliferation, survival, degranulation, or cytokine stimulation following FcεRI triggering, but it did diminish the surface expression of TLR4 and the consequent response to LPS. A similar phenotype was observed in human MCs using an IRE1α inhibitor. Our data indicate that the ER of MCs, primarily of humans, undergoes a rapid remodeling in response to activation that promotes responses to TLR4. We suggest that IRE1α inhibition can be a strategy for inhibiting the hyperactivation of MCs by LPS over the course of allergic responses.


Clinical and biochemical responses to the treatment of milk fever.
One hundred and eighty-six cases of milk fever in 80 herds spread over five counties were used to compare treatment with 8g calcium plus 500 mg magnesium in aspartate with the conventional 12-36g calcium as borogluconate. The data obtained from both the herds and their individual cases show that the treatments gave broadly similar results, and a single intravenous treatment cured 74-7 per cent of cases. It appeared that milk fever was not likely to recur with each succeeding parturition. No breed susceptibility was recognised. No correlation was found between the severity of the clinical signs and the inorganic phosphate concentrations. An incidence of milk fever of 8-79 per cent was recorded.


Uterine rotation: a cause of intestinal obstruction.
Intestinal obstruction is an uncommon surgical emergency during pregnancy that affects seriously the prognosis of gestation. The underlying cause can be identified in the majority of cases and usually consists of adhesions secondary to previous abdominal or pelvic surgery, followed in order of frequency by intestinal volvuli. In recent years there have been no reports in which the gravid uterus has been the cause of intestinal obstruction. We report the case of a woman in week 33 + 4 of pregnancy who developed extrinsic compression of the colon secondary to uterine rotation and pelvic impaction of the head of the fetus.


Multidrug Resistance-associated Protein-1 (MRP-1)-dependent Glutathione Disulfide (GSSG) Efflux as a Critical Survival Factor for Oxidant-enriched Tumorigenic Endothelial Cells.
Endothelial cell tumors are the most common soft tissue tumors in infants. Tumor-forming endothelial (EOMA) cells are able to escape cell death fate despite excessive nuclear oxidant burden. Our previous work recognized perinuclear Nox-4 as a key contributor to EOMA growth. The objective of this work was to characterize the mechanisms by which EOMA cells evade oxidant toxicity and thrive. In EOMA cells, compared with in the cytosol, the nuclear GSSG/GSH ratio was 5-fold higher. Compared to the ratio observed in healthy murine aortic endothelial (MAE) cells, GSSG/GSH was over twice as high in EOMA cells. Multidrug resistance-associated protein-1 (MRP-1), an active GSSG efflux mechanism, showed 2-fold increased activity in EOMA compared with MAE cells. Hyperactive YB-1 and Ape/Ref-1 were responsible for high MRP-1 expression in EOMA. Proximity ligand assay demonstrated MRP-1 and YB-1 binding. Such binding enabled the nuclear targeting of MRP-1 in EOMA in a leptomycin-B-sensitive manner. MRP-1 inhibition as well as knockdown trapped nuclear GSSG, causing cell death of EOMA. Disulfide loading of cells by inhibition of GSSG reductase (bischoloronitrosourea) or thioredoxin reductase (auranofin) was effective in causing EOMA death as well. In sum, EOMA cells survive a heavy oxidant burden by rapid efflux of GSSG, which is lethal if trapped within the cell. A hyperactive MRP-1 system for GSSG efflux acts as a critical survival factor for these cells, making it a potential target for EOMA therapeutics.