Liver X receptors activation, through TO901317 binding, reduces neuroinflammation in Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disease in which degeneration of nigrostriatal neurons and inflammation are key players. The aim of our study was to analyze the function of LXRs in neurodegenerative diseases as PD using in vivo, ex vivo and in vitro models of PD; for this purpose, we observed the effects of the LXR agonist, TO901317, in neuroinflammatory pathway related to PD. We performed an in vivo model of PD using the neurotoxin 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine (MPTP) and our results clearly showed that TO901317 administration reduces all of the inflammatory markers involved in PD such as iNOS and COX2, IκB-α and NF-κB. Moreover, to confirm the neuroprotective properties of TO901317, that we obtained with the in vivo model, we performed also an ex vivo and in vitro models of PD. All the results taken, confirmed that TO901317 is able to modulate the neuroinflammatory pathway involved in PD increasing the locomotors function. Therefore, TO901317, LXR synthetic agonist, could be studied as a new target in a neurodegenerative disorder like PD.


Proteins in vacuo: denaturing and folding mechanisms studied with computer-simulated molecular dynamics.
Mounting evidence from experiments suggests that the native fold in solution is metastable in dehydrated proteins. Results from a number of experiments that use mass spectrometry indicate also that folding-unfolding transitions take place in protein ions even in the absence of water. These observations on anhydrous proteins call for a re-evaluation of our understanding of the folding transition. In this context, computer-assisted simulations are an important complementary tool. Here, we provide an overview of recent progress on the simulation of proteins in vacuo. In particular, we discuss the response of proteins and protein ions to perturbations that trigger unfolding and re-folding transitions. By comparing the general patterns emerging from theory and experiment, we propose a series of new measurements that could help to validate, and improve, current simulation models.


DNA damage as an early biomarker of effect in human health.
In the last few decades the need for new approaches to assess DNA damage has been increasing due to the implications that different insults on genetic material may have on human health. In this context, the identification of how chemical agents with different mechanisms of action (i.e., antineoplastic drugs) damage DNA provides a good model to investigate some cellular and molecular mechanisms underlying the basis of genetic toxicology. The nasal epithelium is the first barrier with which environmental pollutants interact, and for this reason this epithelium can be useful as a sentinel in order to assess the interactions between the environment and the living organisms. Taking these phenomena into account and using a simple, sensitive and rapid method such as the single cell gel electrophoresis, we could obtain information and an initial approach on the DNA status. This assay in combination with other techniques that provide more information about other molecular parameters could give us a better view of the biological status of the living cell.


Rice as a vehicle for dietary fluoride uptake.
The objective of this study was to analyze aspects of the metabolism and bio-availability of fluoride after consumption of a sample of polished white rice containing 5.6 ppm fluoride ion. Up to 400 g of fluoridated rice was consumed by three volunteer adult subjects over specific time periods on two separate occasions. Saliva concentrations were elevated immediately and remained so 90 minutes following ingestion, among other indications. It was concluded that polished rice has the potential to be a useful vehicle for dietary fluoride transport.


Optimal teicoplanin dosage regimens for methicillin-resistant Staphylococcus aureus infections in endocarditis patients and renal failure patients.
This study aimed to assess whether traditional initial loading and maintenance doses of teicoplanin were appropriate in endocarditis and renal failure patients with methicillin-resistant Staphylococcus aureus (MRSA) infections and to recommend optimal dosage regimens. Pharmacokinetic parameters and physicochemical properties of teicoplanin were performed to develop pharmacokinetic models using GastroPlusTM. Concentration-time curves of teicoplanin in endocarditis and renal failure patients with MRSA infections were simulated by changing clearance (CL) and volume of distribution of the central compartment (Vc). Different teicoplanin dosage regimens were assessed according to the target trough concentration, and optimal teicoplanin dosage regimens were recommended. Dosage regimen of four teicoplanin doses of 6 mg/kg q12 h followed by 6 mg/kg qd is recommended for renal failure patients infected by MRSA. And optimal dosage regimen is five teicoplanin doses of 15 mg/kg q12 h followed by doses of 12 mg/kg qd for endocarditis patients infected by MRSA.


Water-induced phase separation of miconazole-poly (vinylpyrrolidone-co-vinyl acetate) amorphous solid dispersions: Insights with confocal fluorescence microscopy.
The aim of this study was to evaluate the utility of confocal fluorescence microscopy (CFM) to study the water-induced phase separation of miconazole-poly (vinylpyrrolidone-co-vinyl acetate) (mico-PVPVA) amorphous solid dispersions (ASDs), induced during preparation, upon storage at high relative humidity (RH) and during dissolution. Different fluorescent dyes were added to drug-polymer films and the location of the dyes was evaluated using CFM. Orthogonal techniques, in particular atomic force microscopy (AFM) coupled with nanoscale infrared spectroscopy (AFM-nanoIR), were used to provide additional analysis of the drug-polymer blends. The initial miscibility of mico-PVPVA ASDs prepared under low humidity conditions was confirmed by AFM-nanoIR. CFM enabled rapid identification of drug-rich and polymer-rich phases in phase separated films prepared under high humidity conditions. The identity of drug- and polymer-rich domains was confirmed using AFM-nanoIR imaging and localized IR spectroscopy, together with Lorentz contact resonance (LCR) measurements. The CFM technique was then utilized successfully to further investigate phase separation in mico-PVPVA films exposed to high RH storage and to visualize phase separation dynamics following film immersion in buffer. CFM is thus a promising new approach to study the phase behavior of ASDs, utilizing drug and polymer specific dyes to visualize the evolution of heterogeneity in films exposed to water.


Generation of a Highly Specific Monoclonal Anti-Infliximab Antibody for Harmonization of TNF-Coated Infliximab Assays.
Determination of infliximab (IFX) serum concentrations has been used for treatment optimization of patients with inflammatory bowel disease. A wide range of enzyme-linked immunosorbent assays (ELISA) exists to quantitate IFX. Most of these assays lack specificity and cross-react with other anti-tumor necrosis factor (TNF) agents. The ability of these IFX assays to detect IFX in complex with antidrug antibodies is not known. The objective of our study was to develop an IFX-specific immunoassay to monitor IFX serum concentrations and to evaluate the impact of antidrug antibodies on the assay performance. A panel of monoclonal antibodies toward IFX (MA-IFX) was generated by hybridoma technology and evaluated to replace the polyclonal antibody in a TNF-coated IFX assay. The selected monoclonal antibody-based (MA-based) IFX ELISA was benchmarked to a clinically validated, reference polyclonal antibody-based (pAb-based) IFX ELISA using 209 inflammatory bowel disease serum samples. Fifty-five MA-IFX were generated and grouped into 9 clusters. Of the 22 monoclonal antibodies tested, MA-IFX6B7 was selected for use in the IFX ELISA and the assay was further optimized. MA-IFX6B7 is a high-affinity (KD = 1.40E-09 mol/L), noninhibitory IgG1 antibody that binds to the Fab fragment of IFX and exhibits no cross-reactivity with other anti-TNF drugs. The linearity of an IFX dose-response curve was demonstrated in the range of 1.2-37.5 ng/mL (R = 0.988). The MA-based assay showed a good Pearson correlation (R = 0.986) and agreement (intraclass correlation coefficient = 0.985) with the pAb-based assay. The MA-based assay detects IFX in complex with nonneutralizing anti-IFX antibodies but not when complexed with neutralizing anti-IFX antibodies. In this study, a highly specific MA-IFX was developed as detection antibody in an ELISA to quantify IFX serum concentrations. The assay was benchmarked to the clinically validated reference pAb-based IFX ELISA.


Unique morphological characteristics of mitochondrial subtypes in the heart: the effect of ischemia and ischemic preconditioning.
Three subsets of mitochondria have been described in adult cardiomyocytes - intermyofibrillar (IMF), subsarcolemmal (SSM), and perinuclear (PN). They have been shown to differ in physiology, but whether they also vary in morphological characteristics is unknown. Ischemic preconditioning (IPC) is known to prevent mitochondrial dysfunction induced by acute myocardial ischemia/reperfusion injury (IRI), but whether IPC can also modulate mitochondrial morphology is not known. Morphological characteristics of three different subsets of adult cardiac mitochondria along with the effect of ischemia and IPC on mitochondrial morphology will be investigated. Mouse hearts were subjected to the following treatments (N=6 for each group): stabilization only, IPC (3x5 min cycles of global ischemia and reperfusion), ischemia only (20 min global ischemia); and IPC and ischemia. Hearts were then processed for electron microscopy and mitochondrial morphology was assessed subsequently. In adult cardiomyocytes, IMF mitochondria were found to be more elongated and less spherical than PN and SSM mitochondria. PN mitochondria were smaller in size when compared to the other two subsets. SSM mitochondria had similar area to IMF mitochondria but their sphericity measures were similar to PN mitochondria. Ischemia was shown to increase the sphericity parameters of all 3 subsets of mitochondria; reduce the length of IMF mitochondria, and increase the size of PN mitochondria. IPC had no effect on mitochondrial morphology either at baseline or after ischemia. The three subsets of mitochondria in the adult heart are morphologically different. IPC does not appear to modulate mitochondrial morphology in adult cardiomyocytes.


[Antimicrobial susceptibilities and serotype distribution of Streptococcus agalactiae strains isolated from pregnant women].
Streptococcus agalactiae (Group B streptococcus, GBS) is an important cause of neonatal morbidity and mortality. The aim of this study was to determine the serotype distribution and antibiotic susceptibility patterns of GBS isolated from pregnant women. A total of 671 pregnant women were screened for vaginal carriage of GBS, and vaginal colonization rate was found to be 14.6%. All GBS isolates were susceptible to penicillin, vancomycin, chloramphenicol and ofloxacin. The rates of GBS resistance to tetracycline, erythromycin and clindamycin were 81.6%, 24.5% and 19.4%, respectively. The serotype distribution of GBS isolates was as follows in order of frequency; serotype III (33.7%), serotype Ib (24.5%), serotype V (18.4%), serotype Ia (7.1%), serotype IV (3.1%) and serotype II (2%). Eleven GBS isolates could not be serotyped by the antisera set used in the study.


The endocannabinoid system and psychiatric disorders.
The present review summarizes the latest information on the role and the pharmacological modulation of the endocannabinoid system in mood disorders and its potential implication in psychotic disorders such as schizophrenia. Reduced functionality might be considered a predisposing factor for major depression, so boosting endocannabinoid tone might be a useful alternative therapeutic approach for depressive disorders. The picture regarding endocannabinoids and anxiety is more complicated since either too much or too little anandamide can lead to anxiety states. However, a small rise in its level in specific brain areas might be beneficial for the response to a stressful situation and therefore to tone down anxiety. This effect might be achieved with low doses of cannabinoid indirect agonists, such as blockers of the degradative pathway (i.e. FAAH) or re-uptake inhibitors. Moreover several lines of experimental and clinical evidence point to a dysregulation of the endocannabinoid system in schizophrenia. The high anandamide levels found in schizophrenic patients, negatively correlated with psychotic symptoms, point to a protective role, whereas the role of 2-arachidonoyl glycerol is still unclear. There is a potential for pharmacological manipulation of the endocannabinoid system as a novel approach for treating schizophrenia, although experimental findings are still controversial, often with different effects depending on the drug, the dose, the species and the model used for simulating positive or negative symptoms. Besides all these limitations, SR141716A and cannabidiol show the most constant antipsychotic properties in dopamine- and glutamate-based models of schizophrenia, with profiles similar to an atypical antipsychotic drug.


[Nocardiosis in patients with human immunodeficiency virus infection].
Nocardiosis is a bacterial disease that occurs in immunocompromised patients, including those infected by the human immunodeficiency virus (HIV). We retrospectively analyzed the clinical records of 27 HIV-positive patients with nocardiosis seen during the period of 1993 to 2004. Clinical presentations, diagnostic methods, predominant species, antimicrobial therapy and outcome were analyzed. Among the total, 81% were males and the median age was 30 years. There was an elevated percentage of alcoholism (89%), smoking (80%) and intravenous drug use (82%). A previous positive serology for HIV infection was present in 85% of the patients. Plasma CD4+ T cell count at the time of diagnosis in 15 of 17 patients (88%) was below 50 cells/microl (median 15 cells/microl). The most frequent clinical onset was pulmonary in 70%, followed by cutaneous in 11% and disseminated in 11%. The main specimens for diagnostic bacterial isolation were sputum (54%), skin and soft tissues (22%) and bronchoalveolar lavage (19%). The predominant pulmonary radiological pattern was alveolar infiltration (74%), followed by cavitations (32%). The species was identified in 13 patients (48%); Nocardia asteroides was isolated in 84% (n = 11). The main antimicrobial drugs prescribed were cotrimoxazole (78%), amikacin (59%) and ciprofloxacin (33%). Dual therapy was used in 78% of the cases, with cotrimoxazole-amikacin being the most frequent. Overall mortality was 37%. Nocardiosis is an unusual infection among HIV-infected patients. The diagnosis should be considered in patients with CD4+ T cell counts below 50/microL and lung or pericardial involvement.


Nephropathy associated with heroin abuse in Caucasian patients.
Renal disease is a complication of heroin addiction. Using renal biopsies in Caucasian patients, we studied the types of nephropathy associated with heroin abuse. Nineteen renal biopsies were performed on heroin addicts between January 1993 and December 2001. The indications for renal biopsy included proteinuria with or without renal insufficiency. All 19 patients had serological evidence of hepatitis C virus (HCV) infection, one had hepatitis B virus surface antigen and three were HIV positive. Thirteen patients (68.4%) were found to have membranoproliferative glomerulonephritis (MPGN), 12 with type I and one with type III. Of the remaining patients, two had chronic interstitial nephritis, two had acute proliferative glomerulonephritis, one had amyloidosis and one had granulomatous glomerulonephritis with interstitial nephritis. No apparent decline in the incidence of renal disease was observed. In this cohort of male Caucasian heroin addicts, HCV-associated MPGN was the most frequent pattern of nephropathy, showing that the nephropathy associated with heroin abuse in Caucasians is not of the focal and segmental glomerulosclerosis type, in contrast to previous reports on African-Americans. This aspect may have important implications for patient management and prognosis.


Leukotrienes play a role in the control of parasite burden in murine strongyloidiasis.
It is clear that leukotrienes mediate inflammatory response; new aspects of leukotriene function have recently been described. In this study, we demonstrate that leukotrienes are key chemical mediators in the control of parasite burdens in mice infected with Strongyloides venezuelensis. High leukotriene levels were detected in the lungs and small intestines of Swiss mice. In infected Swiss mice treated with MK886, a leukotriene synthesis inhibitor, numbers of adult worms, and eggs/g/feces were greater than in infected-only animals. The MK886 treatment inhibited leukotriene B(4) production in the lungs and small intestines, albeit on different postinfection days. Similarly, parasite burdens and eggs/g/feces were greater in 5-lipoxygenase(-/-) mice than in wild-type animals. These observation were confirmed by histopathological study of the duodena. We subsequently observed significant lower numbers of eosinophils and mononuclear cells in the blood, peritoneal cavity fluid, and bronchoalveolar lavage fluid of Swiss mice treated with MK886. In the lung parenchyma of infected animals, MK886 significantly inhibited synthesis of IL-5 at the beginning of infection, whereas levels of IL-12 increased progressively throughout the postinfection period. However, levels of leukotriene C(4), PGE(2), TNF-alpha, IL-3, IL-4, IFN-gamma, and IL-10 were comparable between the treated and untreated groups. Nevertheless, IgE and IgG1 (but not IgG2a) synthesis was also significantly inhibited by MK886 administration. Therefore, in S. venezuelensis-infected mice, adult worm and egg burdens are leukotriene dependent. These findings indicate potential immunostimulatory strategies involving leukotriene administration, and may serve as an alert to physicians treating Strongyloides stercoralis-infected patients presenting asthma-like symptoms because use of 5-lipoxygenase inhibitors may worsen the infection.


Immunogenicity of the human immunodeficiency virus (HIV) recombinant nef gene product. Mapping of T-cell and B-cell epitopes in immunized chimpanzees.
The nonstructural nef gene product of human immunodeficiency virus (HIV), p27, is a regulatory "early phase" protein produced by HIV-infected cells. As a possible negative regulator of transcription, it has been suggested that p27 may be involved in the control of HIV proviral latency. Immune reactivity to p27 may result in early destruction of HIV-replicating cells before viral assembly or of latently infected cells. It appeared, thus, of interest to investigate the immunogenicity of the molecule in chimpanzees immunized against HIV antigens. Two of the six chimpanzees that were injected with soluble recombinant p27 in association with other HIV proteins, displayed significant and sustained T-helper lymphocyte proliferative responses to p27 and to the other antigens. Using a set of synthetic peptides spanning the entire p27 sequence, two T-cell epitopes could be located: one within the last 20 amino-acids of the C terminus of the molecule, the other around the region of residues 118-122. Sera from the same animals also reacted to p27 in a radioimmunoassay as well as to some of the peptides in enzyme-linked immunosorbent assay. Sequential B-cell epitopes could thus be determined as being located in the regions of amino acids: 17-35, 52-66, and 185-205. The results obtained with peptides spanning the region between amino acid residues 65 and 172 indicate that at least two additional B-cell epitopes were present in the region comprised between amino acid 65 and 146. Interestingly, the extreme C terminus of the molecule encompasses both immunodominant T- and B-cell epitopes. Taken together, these observations should prove useful for the rational design of a HIV vaccine.


Ecology and epidemiology of fox rabies.
Although fox rabies has been reported in every country that has foxes, current foci of fox rabies are limited to Europe and North America. The fox rabies virus has unique characteristics that seem to be the result of adaptation through successive passages, i.e., high pathogenicity for the fox, excretion in saliva of a high proportion of infected animals, and low frequency of postinfection immunity. The spread of fox rabies is greatly favored by the characteristics of the genus Vulpes--ubiquity, broad diet, prolific nature, and its particular ethology and ecology. Animals may be immunized by oral vaccination, but natural mechanisms that also can terminate outbreaks are discussed.


Transcriptome analysis reveals in vitro-cultured regeneration bulbs as a promising source for targeted Fritillaria cirrhosa steroidal alkaloid biosynthesis.
The bulbs of Fritillaria cirrhosa is wildly used in traditional Chinese medicine to treat lung-related disease, which has recently been found to have antitussive, anti-inflammatory, antihypertensive and anti-tumor activity. Steroidal alkaloids are the major effective ingredients of F. cirrhosa. In the current study, we demonstrated an efficient strategy for F. cirrhosa bulb regeneration in vitro by cytokinin/auxin induction. Our data showed that the regenerated bulbs accumulated higher alkaloid content that the wild ones. We further performed RNA-seq and bioinformatics analysis to study the gene expression profile, especially those related to alkaloids biosynthesis. KEGG pathway annotation identified genes related to "Metabolic pathways" were the most abundant (2644, 26.0%), followed by those for "Biosynthesis of secondary metabolites" (1319, 13.0%) among the 113,865 unigenes identified. Further analysis suggested MEP pathway, other than MVA pathway, might be the major route for steroidal alkaloid biosynthesis of F. cirrhosa, as all the key genes in this pathway were found to be unregulated in our study. We also showed that accumulation of different phytochemicals was linked to plant hormone addition. Our current study demonstrated that in vitro cultivation is a promising strategy for mass production of F. cirrhosa steroidal alkaloids for pharmacological industry.


The conditional association of problematic drinking with suicidal ideation by alcohol expectancies.
Theory has identified alcohol expectancies as a facilitating factor in the association between problematic drinking and suicidal ideation. In the first test of this question, we explored whether the impact of problematic drinking on suicidal ideation depended on alcohol expectancies. We conducted a secondary analysis of cross-sectional, observational, survey data from a convenience sample of 444 individuals court-ordered to domestic violence intervention programs (69.5% non-Hispanic White; 78.6% Male-identified; Mage = 32.53, SDage = 10.10). Parallel analysis and exploratory factor analysis revealed a three-factor structure of the Effects of Drinking Alcohol Scale, including alcohol expectancies related to 1) disinhibition and negative mood, 2) positive mood, and 3) physical and cognitive effects. Multiple linear regression employing bootstrapping procedures tested the moderating effects of these expectancies on the association between problematic drinking and suicidal ideation, while controlling for gender and depressive symptoms. Problematic drinking was negatively associated with suicidal ideation at low levels of disinhibition and negative mood expectancies, and this association became more positive as these expectancies increased. Problematic drinking was negatively associated with suicidal ideation at high levels of positive mood and became more positive as these expectancies decreased. These preliminary findings suggest that expectancies related to the mood-altering and disinhibiting effects of alcohol may play a role in whether problematic drinking facilitates suicidal thinking. Future intensive longitudinal designs are needed to test whether this moderation is replicated during periods of acute alcohol intoxication and when other psychiatric symptoms are controlled.


Implementing evidence-based practice: reducing risk for low birth weight through pregnancy smoking cessation.
In 1989, the Association of Women's Health, Obstetric and Neonatal Nurses (AWHONN) developed a research utilization program to integrate evidence into practice areas where there were large discrepancies between research evidence and clinical practice. The current program, renamed Research-Based Practice (RBP), uses translational research methods to build from evidence such as that in the Cochrane database and to create protocols for integration of research directly into clinical practice. This article describes the development of the sixth project (RBP6), in which an evidence-based protocol to address smoking in pregnancy was integrated into clinical practice. The protocol includes screening women using descriptive statements and integrates the 5 As (ask, advise, assess, assist, arrange) into prenatal and postpartum care at every visit. By integrating smoking cessation counseling into care, nurses may reduce the risk of low birth weight among pregnant women in both the United States and Canada.


The effects of GnRH analogue (buserelin) or hCG (Chorulon) on Day 12 of pregnancy on ovarian function, plasma hormone concentrations, conceptus growth and placentation in ewes and ewe lambs.
The objectives of this study were to determine the effect of GnRH analogue (buserelin) or human chorionic gonadotrophin (hCG, Chorulon) treatment on Day 12 of pregnancy on ovarian function, plasma hormone concentrations, conceptus growth and placentation in ewes and ewe lambs. After oestrus synchronization with progestagen sponges and eCG, all the animals were mated with fertile rams. Both ewes and ewe lambs (20 per treatment group) were given either normal saline or 4 microg GnRH or 200 IU hCG on Day 12 post-mating. Pre- and post-treatment plasma hormone concentrations were determined in seven pregnant animals per treatment group in samples collected 1h before and 0, 2, 4, 6, 8, 24, 48 and 72 h after treatment. Overall mean progesterone concentrations were higher (P<0.001) in ewes as compared with ewe lambs in saline-treated controls. GnRH or hCG treatment increased (P<0.001) mean plasma progesterone concentrations in both age groups, however, post-treatment concentrations were significantly (P<0.05) higher in ewes than in ewe lambs. Oestradiol concentrations were similar in the two control groups. In ewes, but not in ewe lambs, both GnRH and hCG treatments significantly (P<0.05) increased the mean oestradiol concentrations above pre-treatment levels. Moreover, post-treatment oestradiol concentrations in GnRH- and hCG-treated animals were significantly (P<0.05) higher than those in the saline-treated controls. LH release in response to GnRH treatment was greater (P<0.05) in ewes than in ewe lambs, whereas FSH release in ewes was less (P<0.05) than that of ewe lambs. The effects of GnRH or hCG on conceptus growth and placentation was determined at slaughter on Day 25. In ewes, GnRH treatment increased (P<0.05) luteal weight, amniotic sac width and length, and crown-rump length compared with controls, but had no effect on these parameters in ewe lambs. In ewes, hCG treatment also enhanced (P<0.05) luteal weight, amniotic sac width and length, crown-rump length, embryo weight and number of placentomes as compared with controls. In ewe lambs, there was no difference (P<0.05) between hCG and control groups in luteal weight, embryo weight and amniotic sac width but crown-rump length, amniotic sac length and the number of placentomes forming the placenta were greater (P<0.05). In conclusion, GnRH or hCG treatment on Day 12 of pregnancy can increase ovarian function, conceptus growth and placental attachment in ewes. However, these treatments were less effective in ewe lambs.


A rare case of appendicitis incarcerated in an inguinal hernia.
Amyand's hernia was coined after Claudius Amyand (1660-1740), who was the first to describe the presence of a perforated appendix in a hernial sac and also was the first to perform a successful appendectomy in 1735. It is an exceptionally rare condition in which the hernia itself contains the appendix, which may not necessarily be inflamed. The presence of an inflamed appendix further contributes to the rarity of this case. We report a case of acute appendicitis brought on by its incarceration in the inguinal hernia.


[A case of fowl cholera (Pasteurella multocida) in six-week-old broilers (author's transl)].
An outbreak of fowl cholera in a broiler-house with 6,000 six-week-old broilers is reported. The birds were affected with respiratory disease when they were four weeks old. Eight days after recovery, at an age of six weeks, 50 per cent of the birds were severly ill. The birds were reluctant to move and unable to drink. Four hundred birds died in one day. Post-mortem examination revealed marked swelling of the kidney and arthritis of all the hocks and knee joints in all fifteen birds studied. Pure cultures of Pasteurella multocida were isolated from all joints, the heart, the liver and the kidney. Treatment with chloramphenicol resulted in recovery of the birds which were slaughtered under the supervision of the Veterinary Services of the Ministry of Agriculture and Fisheries in the Netherlands. The disease was not observed in the two other broiler-houses on the same farm, which housed 11,000 and 12,500 broilers of the same age. The findings are discussed.


A Phosphole Oxide Based Fluorescent Dye with Exceptional Resistance to Photobleaching: A Practical Tool for Continuous Imaging in STED Microscopy.
The development of stimulated emission depletion (STED) microscopy represented a major breakthrough in cellular and molecular biology. However, the intense laser beams required for both excitation and STED usually provoke rapid photobleaching of fluorescent molecular probes, which significantly limits the performance and practical utility of STED microscopy. We herein developed a photoresistant fluorescent dye C-Naphox as a practical tool for STED imaging. With excitation using either a λ=405 or 488 nm laser in protic solvents, C-Naphox exhibited an intense red/orange fluorescence (quantum yield ΦF >0.7) with a large Stokes shift (circa 5900 cm(-1) ). Even after irradiation with a Xe lamp (300 W, λex =460 nm, full width at half maximum (FWHM)=11 nm) for 12 hours, 99.5 % of C-Naphox remained intact. The high photoresistance of C-Naphox allowed repeated STED imaging of HeLa cells. Even after recording 50 STED images, 83 % of the initial fluorescence intensity persisted.


Molecular markers and targeted therapy of skin rejection in composite tissue allotransplantation.
Skin rejection remains a major hurdle in reconstructive transplantation. We investigated molecular markers of skin rejection with particular attention to lymphocyte trafficking. Skin biopsies (n = 174) from five human hand transplant recipients were analyzed for rejection, characteristics of the infiltrate and lymphocytic adhesion markers. The cellular infiltrate predominantly comprised CD3+ T cells. CD68, Foxp3 and indoleamine 2, 3-dioxygenase expression and the CD4/CD8 increased with severity of rejection. Lymphocyte adhesion markers were upregulated upon rejection, intercellular adhesion molecule-1 and E-selectin correlated best with severity of rejection. Guided by the findings, a specific E- and P-selectin inhibitor was investigated for its effect on skin rejection in a rat hind limb allotransplant model. While efomycine M (weekly s.c. injection into the graft) alone had no effect, long-term allograft survival was achieved when combined with antithymocyte globulin and tacrolimus (control group without efomycine M rejected at postoperative day [POD] 61 +/- 1). Upregulation of lymphocyte trafficking markers correlates with severity of skin rejection and time after transplantation in human hand transplantation. Blocking E- and P-selectin in the skin holds potential to significantly prolong limb allograft survival.


Purification and characterization of a new factor which restores protein synthesis in a conditionally lethal mutant of Escherichia coli.
The mutant Escherichia coli strain, N4316, has a temperature-sensitive defect in a protein factor required for translation in vitro of bacteriophage f2 RNA. We have purified the normal counterpart of this factor from a wild-type strain, using as an assay its ability to restore the activity of mutant extracts at non-permissive temperature. Our final preparation is free of known initiation, propagation, and release factors, proving that the factor is a new component required for translation. The new factor has a molecular weight of 95000 with preliminary data suggesting a subunit structure. 70% of this protein is found in the soluble-cell fraction, the rest being associated with 70-S ribosomes. Kinetic analyses indicate that the factor acts early in translation. Expression of the defect is highly dependent on the Mg2+ concentration, no temperature-sensitivity being apparent at 15 mM Mg2+. At lower Mg2+ concentrations, the defect is expressed only with natural mRNAs such as f2 RNA, and not with artifical polymers such as poly(U). This specificity suggests that the factor may function in events coded by special sequences in the natural messengers


Environmental Surfaces in Healthcare Facilities are a Potential Source for Transmission of Candida auris and Other Candida Species.
Contaminated surfaces have been implicated as a potential route for dissemination of the emerging multidrug-resistant fungal pathogen Candida auris. In laboratory testing, C. auris and other Candida species persisted for 7 days on moist or dry surfaces. Candida species were recovered frequently from the hospital environment, particularly from moist surfaces. Infect Control Hosp Epidemiol 2017;38:1107-1109.


Further delineation of mandibulofacial dysostosis: Toriello type.
We report a boy who presented with mild mandibulofacial dysostosis, growth retardation with microcephaly, bilateral hearing loss, thoracic deformity with a cardiac valvular lesion and bilateral cryptorchidism. The pattern of malformations differs from the classical Treacher Collins syndrome. We consider it to be mandibulofacial dysostosis (MFD), Toriello type with some additional features.


The Flexible Item Selection Task (FIST): a measure of executive function in preschoolers.
Abstraction and cognitive flexibility were assessed in 197 preschool children at 2, 3, 4, and 5 years of age using the Flexible Item Selection Task, a task adapted from the Visual-Verbal Test (Feldman & Drasgow, 1951). On this new inductive task, children were shown a set of 3 cards and required to select 2 cards that matched each other on 1 dimension (Selection 1) and then to select a different pair of cards that matched each other on another dimension (Selection 2). Thus, 1 of the 3 cards always had to be selected twice according to different dimensions. Two-year-olds failed to understand basic task requirements as assessed by a criterial measure. Three-year-olds did more poorly on Selection 1 than 4- and 5-year-olds (who performed near ceiling), suggesting that 3-year-olds had difficulty with the abstraction component of the task. Four-year-olds did worse than 5-year-olds on Selection 2, suggesting that they had difficulty with the cognitive flexibility component (i.e., difficulty selecting the same card on more than 1 dimension). Results are discussed in terms of the development of executive function.


The distribution of surface antigens during fractionation of mouse liver plasma membranes.
1. Antiserum to purified mouse liver plasma membranes was prepared and the partially purified gamma-globulin antibody fraction was iodinated with (125)I. The reaction of the (125)I-labelled gamma-globulin antibody with isolated mouse liver plasma membranes was studied. 2. The gammaglobulin antibody bound specifically to mouse liver plasma membranes and there was little reaction with mouse liver intracellular membranes or with surface-membrane fractions from either rat liver or pig lymphocytes. 3. ;Light' and ;heavy' mouse liver plasma-membrane subfractions bound similar amounts of gamma-globulin antibody, and this is consistent with a surface origin for the light fraction. 5. Plasma membranes were fractionated by sequential extraction with 50mm-NaHCO(3)-Na(2)CO(3) buffer, pH10.2, containing 10mm-EDTA and aq. 33% (v/v) pyridine. The alkali-soluble and -insoluble fractions and the pyridine-soluble and -insoluble fractions all reacted with the antiserum, and the cross-reactivity among the various fractions and with the total plasma membranes was investigated. 5. The results are discussed in terms of the arrangement of the antigenic determinants within the membrane.


Aortoduodenal fistula.
A primary aortoduodenal fistula is usually associated with an atherosclerotic aortic aneurysm, and a secondary fistula with a leaking anastomotic aortic suture line. Two examples of each are reported. The typical features of a primary fistula are haematemesis or melaena, pain, and a pulsatile abdominal mass; the features of a secondary fistula are haematemesis and melaena with a past history of aortic resection. The initial haemorrhage is rarely fatal: a lag period allows urgent laparotomy. The fistula is diagnosed by dissection of the fourth part of the duodenum from the aorta. The treatment is closure of the duodenum, and resection of an aneurysm if present. Reconstruction is by a graft anastomosed to the aorta proximal to the fistula, if the area is clean, or by an extra anatomical bypass, if the area is heavily contaminated.


National trends in pancreaticoduodenal trauma: interventions and outcomes.
Pancreaticoduodenal trauma (PDT) is associated with substantial mortality and morbidity. In this study, contemporary trends were analysed using national data. The Nationwide Inpatient Sample for 1998-2009 was queried for patients with PDT. Interventions including any operation (Any-Op) and pancreas-specific surgery (PSURG) were identified. Trends in treatment and outcomes were determined [complications, length of stay (LoS), mortality] for the Any-Op, PSURG and non-operative (Non-Op) groups. Analyses included chi-squared tests, Cochran-Armitage trend tests and logistic regression. A total of 27 216 patients (nationally weighted) with PDT were identified. Over time, the frequency of PDT increased by 8.3%, whereas the proportion of patients submitted to PSURG declined (from 21.7% to 19.8%; P = 0.0004) and the percentage of patients submitted to non-operative management increased (from 56.7% to 59.1%; P = 0.01). In the Non-Op group, mortality decreased from 9.7% to 8.6% (P < 0.001); morbidity and LoS remained unchanged at ∼40% and ∼12 days, respectively. In the PSURG group, mortality remained stable at ∼15%, complications increased from 50.2% to 71.8% (P < 0.0001) and LoS remained stable at ∼21 days. For all PDT patients, significant independent predictors of mortality included: the presence of combined pancreatic and duodenal injuries; penetrating trauma, and age >50 years. Having any operation (Any-Op) was associated with mortality, but PSURG was not a predictor of death. The utilization of operations for PDT has declined without affecting mortality, but operative morbidity increased significantly over the 12 years to 2009. The development of an evidence-based approach to invasive manoeuvres and an early multidisciplinary approach involving pancreatic surgeons may improve outcomes in patients with these morbid injuries.


CEP55 Promotes Cell Motility via JAK2⁻STAT3⁻MMPs Cascade in Hepatocellular Carcinoma.
Hepatocellular carcinoma (HCC) is one of the most common malignancies and has a poor prognosis. Novel diagnostic or prognostic biomarkers and potential therapeutic targets for HCC are thus urgently needed. CEP55 plays a crucial role in regulating physical cytokinesis. Whether, and how, CEP55 contributes to HCC development remains unclear. Herein, we demonstrate that CEP55 is abnormally upregulated in HCC tissue, and these high levels of CEP55 are closely related to the poor prognosis of HCC patients. Knockdown of CEP55 expression significantly inhibits HCC cell migration and invasion. We also demonstrate that CEP55 physiologically interacts with JAK2 and promotes its phosphorylation; thus, it is a novel regulator of JAK2⁻STAT3 signaling and its target genes MMP2/9. Finally, blocking JAK2 or STAT3 blunts the stimulation of migration and invasion due to CEP55 overexpression. In summary, our results suggest that CEP55, as an oncogene, promotes HCC cell migration and invasion through regulating JAK2⁻STAT3⁻MMPs signaling.


Free-standing supramolecular hydrogel objects by reaction-diffusion.
Self-assembly provides access to a variety of molecular materials, yet spatial control over structure formation remains difficult to achieve. Here we show how reaction-diffusion (RD) can be coupled to a molecular self-assembly process to generate macroscopic free-standing objects with control over shape, size, and functionality. In RD, two or more reactants diffuse from different positions to give rise to spatially defined structures on reaction. We demonstrate that RD can be used to locally control formation and self-assembly of hydrazone molecular gelators from their non-assembling precursors, leading to soft, free-standing hydrogel objects with sizes ranging from several hundred micrometres up to centimeters. Different chemical functionalities and gradients can easily be integrated in the hydrogel objects by using different reactants. Our methodology, together with the vast range of organic reactions and self-assembling building blocks, provides a general approach towards the programmed fabrication of soft microscale objects with controlled functionality and shape.


Degree of antioxidant protection: a parameter to trace the origin and quality of goat's milk and cheese.
Traceability is an essential tool in reassuring consumers and traders that food is as safe, authentic, and of good quality as expected. Today, food traceability procedures often consist of attached documents and declarations, but scientific parameters that could objectively identify a product would be preferable. Scientific efforts in this area are mostly focused on selection and validation of experimental indicators that would be useful for tracing a food product in any step of its commercial life, describing its raw materials, processing procedures, and storage conditions. In this research, milk and cheese samples from zero grazing and grazing goats were studied to identify a tracing parameter correlated to the feeding system. In particular, alpha-tocopherol and cholesterol were analyzed by HPLC on a normal phase column and were combined to calculate the degree of antioxidant protection (DAP). This parameter, expressed as the molar ratio between antioxidant compounds and an oxidation target, is useful for tracing and distinguishing products from grazing and zero-grazing animals. Degree of antioxidant protection values greater than 7.0 x 10(-3) were found in samples from grazing goats and values lower than 7.0 x 10(-3) were found in samples from zero-grazing goats, for both milk and cheese, meaning that cholesterol was highly protected against oxidative reactions when herbage was the only feed or was dominant in the goat diet. The reliability of DAP to measure the antioxidant protection of cholesterol appeared more effective when the feeding system was based on grazing than when cut herbage was utilized indoors by animals. The DAP index was able to distinguish dairy products when the grazed herbage in the goats' diet exceeded 15%.


A high cell density transient transfection system for therapeutic protein expression based on a CHO GS-knockout cell line: process development and product quality assessment.
Transient gene expression (TGE) is a rapid method for the production of recombinant proteins in mammalian cells. While the volumetric productivity of TGE has improved significantly over the past decade, most methods involve extensive cell line engineering and plasmid vector optimization in addition to long fed batch cultures lasting up to 21 days. Our colleagues have recently reported the development of a CHO K1SV GS-KO host cell line. By creating a bi-allelic glutamine synthetase knock out of the original CHOK1SV host cell line, they were able to improve the efficiency of generating high producing stable CHO lines for drug product manufacturing. We developed a TGE method using the same CHO K1SV GS-KO host cell line without any further cell line engineering. We also refrained from performing plasmid vector engineering. Our objective was to setup a TGE process to mimic protein quality attributes obtained from stable CHO cell line. Polyethyleneimine (PEI)-mediated transfections were performed at high cell density (4 × 10(6) cells/mL) followed by immediate growth arrest at 32 °C for 7 days. Optimizing DNA and PEI concentrations proved to be important. Interestingly, found the direct transfection method (where DNA and PEI were added sequentially) to be superior to the more common indirect method (where DNA and PEI are first pre-complexed). Moreover, the addition of a single feed solution and a polar solvent (N,N dimethylacetamide) significantly increased product titers. The scalability of process from 2 mL to 2 L was demonstrated using multiple proteins and multiple expression volumes. Using this simple, short, 7-day TGE process, we were able to successfully produce 54 unique proteins in a fraction of the time that would have been required to produce the respective stable CHO cell lines. The list of 54 unique proteins includes mAbs, bispecific antibodies, and Fc-fusion proteins. Antibody titers of up to 350 mg/L were achieved with the simple 7-day process. Titers were increased to 1 g/L by extending the culture to 16 days. We also present two case studies comparing product quality of material generated by transient HEK293, transient CHO K1SV GS-KO, and stable CHO K1SV KO pool. Protein from transient CHO was more representative of stable CHO protein compared to protein produced from HEK293.


A major metabolite of delta 1-tetrahydrocannabinol reduces its cataleptic effect in mice.
The results described here demonstrate that THC-induced catalepsy in mice can be substantially inhibited by the prior administration of delta 1-THC-7-oic acid, the major metabolite of THC in most species including humans. This raises the possibility that the intensity and duration of action of THC may depend to a large degree on the levels of this metabolite at the sites of action.


A neurocomputational model of developmental trajectories of gifted children under a polygenic model: When are gifted children held back by poor environments?
From the genetic side, giftedness in cognitive development is the result of contribution of many common genetic variants of small effect size, so called polygenicity (Spain et al., 2016). From the environmental side, educationalists have argued for the importance of the environment for sustaining early potential in children, showing that bright poor children are held back in their subsequent development (Feinstein, 2003a). Such correlational data need to be complemented by mechanistic models showing how gifted development results from the respective genetic and environmental influences. A neurocomputational model of cognitive development is presented, using artificial neural networks to simulate the development of a population of children. Variability was produced by many small differences in neurocomputational parameters each influenced by multiple artificial genes, instantiating a polygenic model, and by variations in the level of stimulation from the environment. The simulations captured several key empirical phenomena, including the non-linearity of developmental trajectories, asymmetries in the characteristics of the upper and lower tails of the population distribution, and the potential of poor environments to hold back bright children. At a computational level, 'gifted' networks tended to have higher capacity, higher plasticity, less noisy neural processing, a lower impact of regressive events, and a richer environment. However, individual instances presented heterogeneous contributions of these neurocomputational factors, suggesting giftedness has diverse causes.


Tumor-Associated Protein Profiles in Kaposi Sarcoma and Mimicking Vascular Tumors, and Their Pathological Implications.
We investigated protein profiles specific to vascular lesions mimicking Kaposi sarcoma (KS), based on stepwise morphogenesis progression of KS. We surveyed 26 tumor-associated proteins in 130 cases, comprising 39 benign vascular lesions (BG), 14 hemangioendotheliomas (HE), 37 KS, and 40 angiosarcomas (AS), by immunohistochemistry. The dominant proteins in KS were HHV8, lymphatic markers, Rb, phosphorylated Rb, VEGF, and galectin-3. Aberrant expression of p53, inactivation of cell cycle inhibitors, loss of beta-catenin, and increased VEGFR1 were more frequent in AS. HE had the lowest Ki-67 index, and the inactivation rates of cell cycle inhibitors in HE were between those of AS and BG/KS. Protein expression patterns in BG and KS were similar. Clustering analysis showed that the 130 cases were divided into three clusters: AS-rich, BG-rich, and KS-rich clusters. The AS-rich cluster was characterized by high caveolin-1 positivity, abnormal p53, high Ki-67 index, and inactivated p27. The KS-rich cluster shared the features of KS, and the BG-rich group had high positive expression rates of galectin-3 and low bcl2 expression. In conclusion, although the rate was different, AS and HE tended to have less cell cycle marker expression than KS, and features of BG and activated KS cell signaling were similar.


Circadian variations in plasma immunoreactive insulin (IRI) and C-peptide concentrations in adult onset (type II) diabetes mellitus.
Plasma immunoreactive insulin (IRI), C-peptide and serum glucose concentrations were determined in 19 adult onset non-insulin dependent (type II) diabetics, in one adult onset diabetic on insulin and in 20 non-diabetic subjects matched for sex, age, weight and height. The subjects lived on a schedule of diurnal activity and nocturnal rest (21:00 to 06:00) at Berceni Clinical Hospital, Bucharest, Romania and ate three meals at 08:30, 13:00 and 18:30 and a snack at 10:00 (carbohydrate content: 50 gm, 65 gm, 60 gm and 25 gm respectively). Nine of the diabetic patients were on oral hypoglycemic agents (tolbutamide or meguan), the others were controlled by diet only. Blood was sampled beginning at 08:00 at 4 hourly intervals over a 24-hour span. The circadian variations of IRI, C-peptide and serum glucose were analysed by population mean cosinor. The non-insulin dependent diabetic subjects as a group and the matched non-diabetic subjects showed under the conditions of this study circadian variations in serum glucose, plasma IRI and C-peptide which were identical in timing (acrophase) and amplitude and with the exception of the much higher serum glucose concentrations in the diabetics, not significantly different in the circadian mean concentration (or mesor). The diabetics on oral hypoglycemic agents if investigated separately showed in spite of identical serum glucose concentration a statistically significantly lower circadian mean IRI and C-peptide concentration than either non-diabetic subjects or the diabetics treated by diet only. Acrophase and relative amplitude remained unchanged. Adult onset non-insulin dependent (type II) diabetics maintained on diet only show the same circadian variations in plasma IRI and C-peptide as non-diabetic matched for sex, age, height and weight. The adult onset diabetic on insulin showed an extremely high serum glucose concentration which varied in its timing over the 24-hr span similar to the other subjects. His plasma IRI concentration was about ten times that of the non-diabetic or other diabetic subjects in this study. There was no statistically recognizable circadian variation of IRI. In contrast C-peptide was found at the same concentration as found in the other two groups and showed in all rhythm parameters an identical circadian variation. The circadian acrophase of plasma IRI and C-peptide concentrations in adult onset diabetics and the non-diabetics is the same.(ABSTRACT TRUNCATED AT 400 WORDS)


Distorted Sandwich α-Diimine PdII Catalyst: Linear Polyethylene and Synthesis of Ethylene/Acrylate Elastomers.
The introduction of m-xylyl substituents to α-diimine PdII catalyst promotes living ethylene polymerization at room temperature and low pressure to yield high molecular weight polyethylene (PE) with low branching (<17/1000 C). m-Xylyl groups provide a highly effective blockage to the axial sites of the catalytic center and form a distorted sandwich geometry. The shielding prevents chain-transfer and easy accessibility of polar monomers, leading to a living polymerization. Conducting a light irradiation as part of the one-step metal-organic insertion light initiated radical (MILRad) process leads to diblock copolymers of ethylene and acrylates. Incorporation of different acrylate block sequences can significantly modify the mechanical and chemical properties of block copolymers which can be modulated to be a hard plastic, elastomer, or semi-amorphous polymer.


Tracheal replacements: Part 2.
In this review, we summarize the history of tracheal reconstruction and replacement as well as progress in current tracheal substitutes. In Part 1, we covered the historical highlights of grafts, flaps, tube construction, and tissue transplants and addressed the progress made in tracheal stenting as a means of temporary tracheal support. In Part 2 we analyze solid and porous tracheal prostheses in experimental and clinical trials and provide a summary of efforts aimed at generating a bioengineered trachea. In both parts, we provide an algorithm on the spectrum of options available for tracheal replacement.


Chlamydia trachomatis infection of the male genital tract: an update.
Chlamydia trachomatis (CT) is the most prevalent cause of sexually transmitted diseases. Although the prevalence of chlamydial infection is similar in men and women, current research and screening are still focused on women, who develop the most severe complications, leaving the study of male genital tract (MGT) infection underrated. Herein, we reviewed the literature on genital CT infection with special focus on the MGT. Data indicate that CT certainly infects different parts of the MGT such as the urethra, seminal vesicles, prostate, epididymis and testis. However, whether or not CT infection has detrimental effects on male fertility is still controversial. The most important features of CT infection are its chronic nature and the presence of a mild inflammation that remains subclinical in most individuals. Chlamydia antigens and pathogen recognition receptors (PRR), expressed on epithelial cells and immune cells from the MGT, have been studied in the last years. Toll-like receptor (TLR) expression has been observed in the testis, epididymis, prostate and vas deferens. It has been demonstrated that recognition of chlamydial antigens is associated with TLR2, TLR4, and possibly, other PRRs. CT recognition by PRRs induces a local production of cytokines/chemokines, which, in turn, provoke chronic inflammation that might evolve in the onset of an autoimmune process in genetically susceptible individuals. Understanding local immune response along the MGT, as well as the crosstalk between resident leukocytes, epithelial, and stromal cells, would be crucial in inducing a protective immunity, thus adding to the design of new therapeutic approaches to a Chlamydia vaccine.


A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Rapid identification of microbes in the bloodstream is crucial in managing septicemia because of its high disease severity, and direct identification from positive blood culture bottles through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can shorten the turnaround time. Therefore, we developed a simple method for rapid microbiological identification from positive blood cultures by using MALDI-TOF MS. We modified previously developed methods to propose a faster, simpler and more economical method, which includes centrifugation and hemolysis. Specifically, our method comprises two-stage centrifugation with gravitational acceleration (g) at 600g and 3000g, followed by the addition of a lysis buffer and another 3000g centrifugation. In total, 324 monomicrobial bacterial cultures were identified. The success rate of species identification was 81.8%, which is comparable with other complex methods. The identification success rate was the highest for Gram-negative aerobes (85%), followed by Gram-positive aerobes (78.2%) and anaerobes (67%). The proposed method requires less than 10 min, costs less than US$0.2 per usage, and facilitates batch processing. We conclude that this method is feasible for clinical use in microbiology laboratories, and can serve as a reference for treatments or further complementary diagnostic testing.


Homo- and heteropolymer self-assembly of recombinant trichocytic keratins.
In the past two decades, keratin biomaterials have shown impressive results as scaffolds for tissue engineering, wound healing, and nerve regeneration. In addition to its intrinsic biocompatibility, keratin interacts with specific cell receptors eliciting beneficial biochemical cues. However, during extraction from natural sources, such as hair and wool fibers, natural keratins are subject to extensive processing conditions that lead to formation of unwanted by-products. Additionally, natural keratins suffer from limited sequence tunability. Recombinant keratin proteins can overcome these drawbacks while maintaining the desired chemical and physical characteristics of natural keratins. Herein, we present the bacterial expression, purification, and solution characterization of human hair keratins K31 and K81. The obligate heterodimerization of the K31/K81 pair that results in formation of intermediate filaments is maintained in the recombinant proteins. Surprisingly, we have for the first time observed new zero- and one-dimensional nanostructures from homooligomerization of K81 and K31, respectively. Further analysis of the self-assembly mechanism highlights the importance of disulfide crosslinking in keratin self-assembly.


A collaborative approach to facilitate professionals to support the breathless patient.
Breathlessness is a major problem for people in their last weeks of life. Breathlessness is considered to be multidimensional with physical, psychological, emotional, social and spiritual factors all playing a part. It has been recognised that specific training to health professionals is beneficial in order to improve the care for patients with breathlessness.Breathlessness courses have tended to focus on senior nurses. A new flexible and collaborative training course was designed to include a wider range of nurses and other health professionals in hospital, hospice, primary care and community settings. The aim of the 'Practical Skills to Support the Breathless Patient' programme was to make patients and carers feel better supported in their breathlessness, for health professionals to develop confidence and skills in using proven interventions, and to adopt a flexible educational design that could be adapted to different contexts. The course is learner-centred and teaching methods encourage interaction and participation via a mix of lectures and discussions with practical skills-focused, experiential workshops in smaller groups. Case study work was included to integrate learning with participants' practice environment. Evaluation is built in during the course, so adaptations can be made throughout to respond to changing learner needs. Participants reported increased confidence in terms of knowledge and applying this within everyday practice. The theory-practice dynamic worked well within each participant' specific work context in particular through the case study approach. The course developed a number of innovative approaches, such as multi-disciplinary learning groups, regular feedback loops, reflexive learning about putting theory into practice and long-term follow-up. Combining these elements increases professionals' confidence and sustains new clinical practice.


Unique conformation of cancer autoantigen B23 in hepatoma: a mechanism for specificity in the autoimmune response.
The association of a specific autoantibody response with distinct disease phenotypes is observed in both autoimmune diseases and cancer. Although the underlying mechanisms remain unclear, it is likely that unique properties of disease-specific autoantigens expressed in the relevant target cells play a role. It has recently been observed that the majority of autoantigens targeted across the spectrum of systemic autoimmune diseases (but not nonautoantigens) are selectively cleaved by the cytotoxic lymphocyte granule protease granzyme B (GB), generating unique fragments not observed during other forms of cell death. Although susceptibility of a molecule to cleavage by GB strongly predicts autoantigen status, the significance of this association is unclear. We used hepatocellular carcinoma and the hepatocellular carcinoma autoantigen, nucleophosmin/B23, as a model system to define the unique features of disease-specific autoantigens in the relevant disease microenvironment. These studies revealed a striking, selective susceptibility of B23 to cleavage by GB in extracts of neoplastic liver. The increased sensitivity of tumor B23 to proteolysis by GB was accompanied by slightly increased mobility on SDS/PAGE, altered subcellular localization, enrichment of an SDS-stable oligomeric form of B23, and recognition by a conformation-specific antibody detecting a B23 epitope ending at the GB cleavage site. In vitro studies demonstrated that this unique B23 conformation and resultant increased susceptibility to cleavage by GB arise when B23 translation is initiated at methionine-7. We propose that unique features of autoantigens in the disease-relevant microenvironment may regulate susceptibility to cleavage by GB and their selection by the specific autoimmune response.


Regulatory effects of baicalin, a flavonoid compound, on adipocyte metabolism.
Baicalin is a plant-derived, biologically active compound exerting numerous advantageous effects. Adipocytes store and release energy in the process of lipogenesis and lipolysis. Rodent studies have shown that baicalin treatment positively affects fat tissue, however, data on the direct influence of this compound on adipocyte metabolism is lacking. In the present research, the short-term effects of 25, 50, and 100 μM baicalin on glucose transport, conversion to lipids, and oxidation, and also on lipolysis in primary rat adipocytes were explored. Lipolysis was measured as glycerol release from adipocytes. It was shown that 100 μM baicalin reduced glucose oxidation but at any concentration did not affect glucose transport and lipogenesis. Baicalin significantly increased the adipocyte response to physiological and pharmacological lipolytic stimuli (such as epinephrine - adrenergic agonist, DPCPX - adenosine A1 receptor antagonist, and amrinone - cAMP phosphodiesterase inhibitor). The stimulatory effects of baicalin on epinephrine-induced lipolysis were markedly diminished by insulin (activator of cAMP phosphodiesterases) and H-89 (PKA inhibitor). It was also demonstrated that baicalin evoked a similar rise in epinephrine-induced lipolysis in the presence of glucose and alanine. Our results provided evidence that baicalin may reduce glucose oxidation and is capable of enhancing lipolysis in primary rat adipocytes. The action on lipolysis is glucose-independent and covers both the adrenergic and adenosine A1 receptor pathways. The rise in cAMP content is proposed to be responsible for the observed potentiation of the lipolytic process.


Association Between Environmental Factors and Oral Epstein-Barr Virus DNA Loads: A Multicenter Cross-sectional Study in China.
Oral Epstein-Barr virus (EBV) status reflects host EBV activity and potentially links to EBV-associated diseases, however, factors influencing oral EBV loads or reactivation, such as environmental exposures or host factors, are not fully understood. A 2-stage, multicenter, cross-sectional study of 6558 subjects from 21 administrative cities of southern China and 3 populations from representative geographical areas in China (referred to as the south, north, and northeastern populations) was performed. The relationships between demographical factors and environmental exposures to EBV loads were analyzed by logistic regression models. Current smoking, with a dose-response effect, was found to be strongly associated with higher oral EBV loads in the pooled data, with an odds ratio of 1.58 (95% confidence interval, 1.39-1.79), as well as in each of the separate populations. The odds ratio increased to 3.06 when current smokers in southern China were compared to never smokers in northern China. Additionally, higher oral EBV loads tended to be detected in older participants, male participants, and participants in southern China. This study provided evidence linking the effect of host-environmental factors, particularly smoking, to oral EBV activity. It could strengthen our understanding of the possible causal roles of EBV-related diseases, which may help to prevent or mitigate EBV-associated diseases.


Darunavir concentrations in CSF of HIV-infected individuals when boosted with cobicistat versus ritonavir.
Cobicistat and ritonavir have different inhibitory profiles for drug transporters that could impact the distribution of co-administered drugs. We compared darunavir concentrations in CSF when boosted by cobicistat versus ritonavir relative to plasma concentrations and with WT HIV-1 IC50 and IC90. An open, single-arm, sequential clinical trial (NCT02503462) where paired CSF and blood samples were taken from seven HIV-infected patients presenting with HIV-associated neurocognitive disorders (HAND) and treated with a darunavir/ritonavir (800/100 mg) once-daily regimen. Ritonavir was subsequently replaced by cobicistat and paired CSF and blood samples were obtained from the same patients after treatment with the darunavir/cobicistat (800/150 mg) once-daily regimen. Darunavir concentrations at the end of the dosing interval were quantified by LC-MS/MS. The median (IQR) darunavir concentrations in CSF with ritonavir and cobicistat boosting were 16.4 ng/mL (8.6-20.3) and 15.9 ng/mL (6.7-31.6), respectively (P = 0.58). The median (IQR) darunavir CSF:plasma ratios with ritonavir and cobicistat boosting were 0.007 (0.006-0.012) and 0.011 (0.007-0.015), respectively (P = 0.16). Darunavir concentrations in CSF exceeded the darunavir IC50 and IC90 by a median of 9.2- and 6.7-fold with ritonavir boosting, and by 8.9- and 6.5-fold with cobicistat boosting, respectively. All patients had darunavir CSF concentrations above the target inhibitory concentrations and remained virologically suppressed in the CSF and plasma. This small study shows that cobicistat and ritonavir give comparable effective darunavir concentrations in CSF, thus suggesting that these boosters can be used interchangeably in once-daily darunavir regimens.


Treatment of adolescent idiopathic scoliosis using Texas Scottish Rite Hospital instrumentation.
To determine the effectiveness of posterior TSRH instrumentation for the treatment of idiopathic scoliosis, 103 patients with a 2-year minimum followup were retrospectively studied. Patients who underwent operations between October 1988 and April 1991 were evaluated for curve correction, spinal balance, and complications. Age at surgery averaged 14.3 years. Follow-up averaged 2.5 years. Thoracic curve correction averaged 65% in those with King Type III/IV curves and 54% in those with Type II curves. With follow-up, correction loss averaged approximately 13% for each group. Lumbar curve correction after instrumentation in Type I and II curves averaged 48% postop but lost approximately 20% with follow-up. Trunk balance improved 77% toward midline after surgery in those with Type III/IV curves. Improvement in trunk balance was less impressive in patients with Type II curves, particularly after selective thoracic fusions. Thoracic sagittal contour improved 43% for hypokyphotic (< 20 degrees) patients but, in the remainder, no significant radiographic change was evident. No neurologic complications occurred. Delayed deep infections developed in ten patients (10%) between 11 and 45 months postoperative. Cultures eventually grew Propionibacterium acnes, staph epidermidis, or staph coagulase negative in eight patients. Two patients had pseudarthroses. Frontal and sagittal thoracic curve correction can be satisfactorily obtained using TSRH instrumentation. Continued efforts are being made to improve lumbar hook patterns and technique to achieve and maintain better lumbar curve correction.


Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of ε-Acetamidocaproic Acid in Rat Plasma.
A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of ε-acetamidocaproic acid (AACA), the primary metabolite of zinc acexamate (ZAC), in rat plasma by using normetanephrine as an internal standard. Sample preparation involved protein precipitation using methanol. Separation was achieved on a Gemini-NX C18 column (150 mm × 2.0 mm, i.d., 3 μm particle size) using a mixture of 0.1% formic acid-water : acetonitrile (80 : 20, v/v) as the mobile phase at a flow rate of 200 μl/min. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 4.0 min, and the calibration curves of AACA were linear over the concentration range of 20~5000 ng/ml in rat plasma. The coefficient of variation and relative error at four QC levels were ranged from 1.0% to 5.8% and from -8.4% to 6.6%, respectively. The present method was successfully applied for estimating the pharmacokinetic parameters of AACA following intravenous or oral administration of ZAC to rats.


Population distribution and burden of acute gastrointestinal illness in British Columbia, Canada.
In developed countries, gastrointestinal illness (GI) is typically mild and self-limiting, however, it has considerable economic impact due to high morbidity. The magnitude and distribution of acute GI in British Columbia (BC), Canada was evaluated via a cross-sectional telephone survey of 4,612 randomly selected residents, conducted from June 2002 to June 2003. Respondents were asked if they had experienced vomiting or diarrhoea in the 28 days prior to the interview. A response rate of 44.3% was achieved. A monthly prevalence of 9.2% (95% CI 8.4-10.0), an incidence rate of 1.3 (95% CI 1.1-1.4) episodes of acute GI per person-year, and an average probability that an individual developed illness in the year of 71.6% (95% CI 68.0-74.8), weighted by population size were observed. The average duration of illness was 3.7 days, translating into 19.2 million days annually of acute GI in BC. The results corroborate those from previous Canadian and international studies, highlighting the substantial burden of acute GI.


Enhancing Culturally and Linguistically Responsive Features of Individualized Education Programs (IEPs) for Speech-Language Pathologists (SLPs).
Individual education programs (IEPs) are the foundation for guiding speech-language pathologists (SLPs) to develop appropriate interventions for culturally and linguistically diverse (CLD) students who qualify for speech-language services under the Individuals with Disabilities Education Act. There is a growing number of CLD students with speech-language impairments who need special attention given to their culture and language. This article highlights key culturally and linguistically responsive features to aid SLPs in developing meaningful IEPs for this diverse group of students.


The effect of metabolic control on rate of decline in renal function in insulin-dependent diabetes mellitus with overt diabetic nephropathy.
Strict glycaemic control has been shown to reduce the risk of developing diabetic nephropathy. In established nephropathy, however, the impact of glycaemic control on prognosis is less clear. Therefore we investigated the effect of long-term glycaemic control on the decline in renal function in insulin-dependent diabetic patients with overt nephropathy. The study was performed at two hospital-based diabetes centres in western Sweden. The study was an observational retrospective follow-up study in 158 insulin-dependent diabetics with proteinuria with a mean (+/-SD) age of 36+/-9 years and a diabetes duration of 22+/-8 years. The change in glomerular filtration rate was measured as 51Cr EDTA clearance for a median of 8 years (range 1-17). Glycaemic control was determined with measurements of glycated haemoglobin A1c. The decline in glomerular filtration rate was 3.8+/-3.7 ml/min/year. The blood pressure was 143/82+/-15/7 mmHg and the mean glycated haemoglobin was 8.7+/-1.6%. The correlation coefficient between glycated haemoglobin and decline in glomerular filtration rate was -0.39 (P<0.0001 ) and between decline in glomerular filtration rate and systolic and diastolic blood pressure -0.17 (P=0.03) and -0.29 (P=0.003) respectively. In patients with glycated haemoglobin <8.0% and diastolic blood pressure <85 mmHg the decline in glomerular filtration rate was 1.7+/-2.3 ml/min/year. In this retrospective observational study, effective blood-pressure control was associated with a low rate of decline in renal function and a low urinary albumin excretion. The correlation between glycaemic control and decline in renal function indicates that poor glycaemic control can accelerate the loss of renal function in diabetic nephropathy.


Minor physical anomalies and quantitative measures of the head and face in patients with psychosis.
The aim of this study was to examine minor physical anomalies and quantitative measures of the head and face in patients with psychosis vs healthy controls. Based on a comprehensive prevalence study of psychosis, we recruited 310 individuals with psychosis and 303 controls. From this sample, we matched 180 case-control pairs for age and sex. Individual minor physical anomalies and quantitative measures related to head size and facial height and depth were compared within the matched pairs. Based on all subjects, we examined the specificity of the findings by comparing craniofacial summary scores in patients with nonaffective or affective psychosis and controls. The odds of having a psychotic disorder were increased in those with wider skull bases (odds ratio [OR], 1.40; 95% confidence interval [CI], 1.02-1.17), smaller lower-facial heights (glabella to subnasal) (OR, 0.57; 95% CI, 0.44-0.75), protruding ears (OR, 1.72; 95% CI, 1.05-2.82), and shorter (OR, 2.29; 95% CI, 1.37-3.82) and wider (OR, 2.28; 95% CI, 1.43-3.65) palates. Compared with controls, those with psychotic disorder had skulls that were more brachycephalic. These differences were found to distinguish patients with nonaffective and affective psychoses from controls. Several of the features that differentiate patients from controls relate to the development of the neuro-basicranial complex and the adjacent temporal and frontal lobes. Future research should examine both the temporal lobe and the middle cranial fossa to reconcile our anthropomorphic findings and the literature showing smaller temporal lobes in patients with schizophrenia. Closer attention to the skull base may provide clues to the nature and timing of altered brain development in patients with psychosis.


Efficacy of low-dose intravenous cyclophosphamide in systemic lupus erythematosus presenting with Guillain-Barre syndrome-like acute axonal neuropathies: report of two cases.
There are few cases of Guillain-Barré syndrome (GBS), particularly of atypical variants, occurring in association with systemic lupus erythematous (SLE). Reports addressing a specific therapy thus remain almost anecdotal. It is therefore challenging to determine the treatment that is best suited for this subset of patients, especially if initial conventional therapy for GBS fails. We present two cases of GBS-like acute axonal neuropathies, one with acute motor axonal neuropathy (AMAN), and another with acute motor sensory axonal neuropathy (AMSAN), presenting early in the course of SLE. The first case failed to respond to therapy with intravenous immunoglobulins (IVIG) and plasmapheresis, but achieved a favorable outcome when high-dose glucocorticoids along with low-dose intravenous (IV) cyclophosphamide pulses were given. The second case responded favorably to high-dose glucocorticoids, IVIG, and low-dose IV cyclophosphamide pulses. Both patients have remained in clinical remission and without neurologic sequelae after 10 and three years of follow-up, respectively.


[Importance of PPSB fraction in the treatment of hemophilia A with antibodies against factor VIII].
On the basis of the indirectly established statement that activated forms of the coagulation factor are also present in PPSB fractions of own production, the activated concentrate of factor IX and of the prothrombin complex were applied in haemophilia-A patients with antibodies against factor VIII. The fraction was administered to 4 patients during 14 bleeding times, mostly during bleedings of joints an soft tissues, twice during haematuria in a dose of 40-200 E-factor IX per kg of body weight and per day. The total dose was mainly administered in a fractionized way at an interval of 8 or 12 hours. This treatment lasted for 2 to 7 days. With regard to haemostasis no fundamental improvement of global blood coagulation tests (which have pathological results in haemophilia-A patients) could be identified in the course of the treatment. However, the increase of factor II, VII, and X in the patient's plasma was striking. Contrary to exceptations, there was a less distinct increase of factor IX activity. Concluding from these findings it may be assumed that the "strengthening" of the "extrinsic system" is decisive for the haemostatic effect in the treatment mentioned.


Management of periorbital basal cell carcinoma with orbital invasion.
Basal cell carcinoma (BCC) is the most common eyelid malignancy; however, orbital invasion by periocular BCC is rare, and management remains challenging. Established risk factors for orbital invasion by BCC include male gender, advanced age, medial canthal location, previous recurrences, large tumor size, aggressive histologic subtype and perineural invasion. Management requires a multidisciplinary approach with orbital exenteration remaining the treatment of choice. Globe-sparing treatment may be appropriate in selected patients and radiotherapy and chemotherapy are often used as adjuvant therapies for advanced or inoperable cases, although the evidence remains limited. We aim to summarize the presentation and treatment of BCC with orbital invasion to better guide the management of this complex condition.


The potential of health literacy to address the health related UN sustainable development goal 3 (SDG3) in Nepal: a rapid review.
Health literacy has been linked to health outcomes across population groups around the world. Nepal, a low income country, experiences the double burden of highly prevalent communicable as well as non-communicable diseases. The World Health Organization (WHO) has positioned health literacy as a key mechanism to meet the health-related Sustainable Development Goal (SDG3). However, there is little known about the status of health literacy in developing countries such as Nepal. This paper aims to review the potential of health literacy to address SDG3 in Nepal. A rapid review was conducted using the knowledge to action evidence summary approach. Articles included in the review were those reporting on barriers to health care engagements in Nepal published in English language between January 2000 and December 2015. Barriers for healthcare engagement included knowledge and education as strong factors, followed by culture, gender roles, quality of service and cost of services. These barriers influence the Nepalese community to access and engage with services, and make and enact healthcare decisions, not only at the individual level but at the family level. These factors are directly linked to health literacy. Health literacy is a pivotal determinant of understanding, accessing and using health information and health services, it is important that the health literacy needs of the people be addressed. Locally identified and developed health literacy interventions may provide opportunities for systematic improvements in health to address impediments to healthcare in Nepal. Further research on health literacy and implementation of health literacy interventions may help reduce inequalities and increase the responsiveness of health systems which could potentially facilitate Nepal to meet the sustainable development goals. While there is currently little in place for health literacy to impact on the SDG3, this paper generates insights into health literacy's potential role.


Effects of Plasma-Activated Water Treatment on the Inactivation of Microorganisms Present on Cherry Tomatoes and in Used Wash Solution.
Herein, we investigated the potential of plasma-activated water (PAW) as a wash solution for the microbial decontamination of cherry tomatoes. We analyzed the efficacy of PAW as a bactericidal agent based on reactive species and pH. Immersion for 5 min in PAW15 (generated via plasma activation for 15 min) was determined as optimal for microbial decontamination of fresh produce. The decontamination efficacy of PAW15 exceeded those of mimic solutions with equivalent reactive species concentrations and pH (3.0 vs. 1.7 log reduction), suggesting that the entire range of plasma-derived reactive species participates in decontamination rather than a few reactive species. PAW15-washing treatment achieved reductions of 6.89 ± 0.36, 7.49 ± 0.40, and 5.60 ± 0.05 log10 CFU/g in the counts of Bacillus cereus, Salmonella sp., and Escherichia coli O157:H7, respectively, inoculated on the surface of cherry tomatoes, with none of these strains detected in the wash solution. During 6 days of 25 °C storage post-washing, the counts of aerobic bacteria, yeasts, and molds were below the detection limit. However, PAW15 did not significantly affect the viability of RAW264.7 cells. These results demonstrate that PAW effectively inactivates microbes and foodborne pathogens on the surface of cherry tomatoes and in the wash solution. Thus, PAW could be used as an alternative wash solution in the fresh produce industry without cross-contamination during washing and environmental contamination by foodborne pathogens or potential risks to human health.


Imaging and scattering studies of the unimolecular dissociation of the BrCH2CH2O radical from BrCH2CH2ONO photolysis at 351 nm.
We report a study of the unimolecular dissociation of BrCH2CH2O radicals produced from the photodissociation of BrCH2CH2ONO at 351/355 nm. Using both a crossed laser-molecular beam scattering apparatus with electron bombardment detection and a velocity map imaging apparatus with tunable VUV photoionization detection, we investigate the initial photodissociation channels of the BrCH2CH2ONO precursor and the subsequent dissociation of the vibrationally excited BrCH2CH2O radicals. The only photodissociation channel of the precursor we detected upon photodissociation at 351 nm was O-NO bond fission. C-Br photofission and HBr photoelimination do not compete significantly with O-NO photofission at this excitation wavelength. The measured O-NO photofission recoil kinetic energy distribution peaks near 14 kcal/mol and extends from 5 to 24 kcal/mol. There is also a small signal from lower kinetic energy NO product (it would be 6% of the total if it were also from O-NO photofission). We use the O-NO photofission P(ET) peaking near 14 kcal/mol to help characterize the internal energy distribution in the nascent ground electronic state BrCH2CH2O radicals. At 351 nm, some but not all of the BrCH2CH2O radicals are formed with enough internal energy to unimolecularly dissociate to CH2Br + H2CO. Although the signal at m/e = 93 (CH2Br(+)) obtained with electron bombardment detection includes signal both from the CH2Br product and from dissociative ionization of the energetically stable BrCH2CH2O radicals, we were able to isolate the signal from CH2Br product alone using tunable VUV photoionization detection at 8.78 eV. We also sought to investigate the source of vinoxy radicals detected in spectroscopic experiments by Miller and co-workers ( J. Phys. Chem. A 2012 , 116 , 12032 ) from the photodissociation of BrCH2CH2ONO at 351 nm. Using velocity map imaging and photodissociating the precursor at 355 nm, we detected a tiny signal at m/e = 43 and a larger signal at m/e = 15 that we tentatively assign to vinoxy. An underlying signal in the time-of-flight spectra at m/e = 29 and m/e = 42, the two strongest peaks in the literature electron bombardment mass spectrum of vinoxy, is also apparent. Comparison of those signal strengths with the signal at HBr(+), however, shows that the vinoxy product does not have HBr as a cofragment, so the prior suggestion by Miller and co-workers that the vinoxy might result from a roaming mechanism is contraindicated.


The effect of tamoxifen on the genital tract.
Tamoxifen is a selective estrogen receptor modulator (SERM) that is widely used in the treatment of patients with breast cancer and for chemoprophylaxis in high risk women. Tamoxifen results in a spectrum of abnormalities involving the genital tract, the most significant being an increased incidence of endometrial cancer and uterine sarcoma. This article reviews the effects of tamoxifen on the genital tract and the strengths and weaknesses of various imaging modalities for evaluating the endometrium.


Novel activation step required for transcriptional competence of progesterone receptor on chromatin templates.
To elucidate the earliest molecular steps in the activation of transcription by the progesterone receptor (PR), we investigated its activity in a cell-free transcription system utilizing chromatin templates. PR prepared as a ligand-free, recombinant protein failed to induce transcription on chromatin templates. However, transcriptional competence could be restored by coincubation with rabbit reticulocyte lysate (RRL). The interaction of PR with chaperones results in a receptor conformation competent to bind ligand and RRL contains abundant chaperone-mediated protein folding activity. Blocking this activity with the specific inhibitor geldanamycin inhibited receptor-dependent transcriptional activity. However, recombinant chaperones could not replace RRL in the restoration of transcriptional activity on chromatin templates, suggesting the presence of an additional activity in the lysate. Under chromatin assembly conditions, PR could bind naked DNA and RRL did not increase that binding. In contrast, PR bound to a chromatin template only poorly. Interestingly, RRL stimulated sequence-specific binding by PR to target sites in chromatin and the concomitant recruitment of the steroid receptor coactivator 1 to the promoter. Thus, our results indicate that a novel protein-mediated activity in RRL is involved in an additional, heretofore unrecognized, activation step required for PR to become transcriptionally competent on chromatin templates.


Influence of CYP2D6 genotype on metoprolol plasma concentration and beta-adrenergic inhibition during long-term treatment: a comparison with bisoprolol.
In patients routinely treated with metoprolol, influences of CYP2D6 genotype on the response of heart rate to isoproterenol (IP) were studied at its peak and trough concentrations and were compared with those of bisoprolol. In 72 patients treated with metoprolol or bisoprolol, CYP2D6 genotype (ie, CYP2D6*1, *2, *4, *5, *10, and *14) was determined. No patients except one who was heterozygous for CYP2D6*5 carried the null alleles of CYP2D6. The homozygote frequency for CYP2D6*10 was relatively high (19.4%) and these patients had greater peak and trough plasma concentrations of metoprolol than the other patients. Isoproterenol-induced percentage increases in heart rate were 58% and 38% less at the low and high rate of isoproterenol infusion (0.02 and 0.04 microg/kg/min), respectively, in patients homozygous for CYP2D6*10 than in the other patients at the trough, but not at the peak concentrations. In contrast, CYP2D6 genotype did not affect plasma concentrations of bisoprolol and the extent of its beta-adrenergic inhibition. Thus, in patients routinely treated with metoprolol, CYP2D6 genotype significantly affects circadian variations of beta-adrenergic inhibition induced by metoprolol. In contrast, bisoprolol has a relatively constant beta-adrenergic inhibition independent of CYP2D6 genotype.


[The effect of oxymatrine on aging mice caused by D+ -galactose].
To investigate the effect of oxymatrine (OMT) on aging mouse caused by D (+)-Galactose. Healthy Kunming mouse were randomly divided into control, model and treated groups. The treated group was divided into three subgroups which were given oxymatrine 5 mg/kg, 10 mg/kg,20 mg/kg (OMT5, OMT10, OMT20). After 30 days of administration,the stand-jumping test and the Morris water maze test of each group were used to observe the learning and memory ability. After the behavior indicators test, 1-2 mL blood were got from ophthalmic vein plexus,and then got the serum by centrifuge. The super oxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined by SOD and MDA kit. In the water maze test, the model group was significantly longer in the latency than that of the control group (P < 0.01), and the number of the crossing platforms was significantly reduced (P < 0.01), but compared with the model group, the OMT5, OMT10, OMT20 groups were significantly shorter in the latency to the end (P < 0.05, P < 0.05, P < 0.01), and the number of the crossing platforms was significantly increased (P < 0.05, P < 0.05, P < 0.01); In the stand-jumping test,the OMT groups all could extend the platform latency and reduce the error reaction times, the learning ability of the OMT groups was better than the model group, but the statistical tests failed (P > 0.05); In the serum biochemical indicators test,the SOD activity of the model group was significantly lower than that of the control group (P < 0.01), but the SOD activity of OMT5, OMTIO, OMT20 groups was significant higher than that of the model group (P < 0.05, P < 0.05, P < 0.01); The MDA content of the model group was significantly higher than that of the control group (P < 0.01) but the MDA content of OMT5, OMT1O, OMT20 groups was significant lower than that of the model group (P < 0.05, P < 0.05, P < 0.01). Oxymatrine could improve the learning and memory ability of aging mice caused by intraperitoneal injection D(+)-Galactose,and the anti-aging effect is related to the oxygen free radicals and reducing the lipid peroxidation.


Differential glycosylation of auditory and vestibular hair bundle proteins revealed by peanut agglutinin.
Up to four morphologically distinct types of cross-link are found between the stereocilia in the hair bundles of avian hair cells. These links are involved in mechanotransduction, force transmission across the bundle, and maintenance of hair bundle structure. They appear to be specialisations of the cell coat, but very little is known about their molecular composition. Chick inner ear tissues were therefore screened with a number of different lectins to find markers for specialisations of the hair bundle surface. One lectin, peanut agglutinin (PNA), which recognises the dissacharide Gal beta 1-3GalNAc, was found to be a fairly selective marker for vestibular hair bundles, but it does not stain the stereocilia of auditory hair cells. The staining patterns observed with PNA in the vestibular system closely resemble those seen with a monoclonal antibody (mab) directed against a 275 kD component of the hair cell's apical surface known as the hair-cell antigen (HCA). However, unlike PNA, the mab recognises both vestibular and auditory hair cells. A detailed comparison of the fluorescence staining patterns observed with PNA and the anti-HCA mab indicates that binding sites for both ligands spatially codistribute on the surface of vestibular hair cells. The lectin and the anti-HCA mab binding sites are both sensitive to trypsin treatment, and, with sections of the vestibular system, PNA pretreatment blocks subsequent anti-HCA mab staining. Immunoelectron microscopy of vestibular hair bundles shows that PNA and the anti-HCA mab both label a type of cross-link known as the shaft connector. This link type is present on both auditory and vestibular hair bundles but reacts with PNA only in the vestibular system. The lectin jacalin, which has greater specificity for Gal beta 1-3GalNAc than does PNA, also only labels vestibular and not auditory hair bundles. Although terminal sialic acid residues can block both PNA and jacalin binding, neuraminidase treatment does not unmask cryptic binding sites for these lectins on auditory hair cells but does reveal PNA and jacalin staining at a number of other locations in the inner ear. The results obtained with the lectins PNA and jacalin indicate that either the HCA or other components of the shaft links are differentially glycosylated in the vestibular and auditory epithelia of the bird. The functional significance for such a difference in glycosylation remains to be determined, but auditory and vestibular hair cells operate over different frequency ranges, and variations in glycosylation might confer different micromechanical properties on the hair bundles in these two systems.


Omics profiles used to evaluate the gene expression of Exiguobacterium antarcticum B7 during cold adaptation.
Exiguobacterium antarcticum strain B7 is a Gram-positive psychrotrophic bacterial species isolated in Antarctica. Although this bacteria has been poorly studied, its genome has already been sequenced. Therefore, it is an appropriate model for the study of thermal adaptation. In the present study, we analyzed the transcriptomes and proteomes of E. antarcticum B7 grown at 0°C and 37°C by SOLiD RNA-Seq, Ion Torrent RNA-Seq and two-dimensional difference gel electrophoresis tandem mass spectrometry (2D-DIGE-MS/MS). We found expression of 2,058 transcripts in all replicates from both platforms and differential expression of 564 genes (absolute log2FC≥1, P-value<0.001) comparing the two temperatures by RNA-Seq. A total of 73 spots were differentially expressed between the two temperatures on 2D-DIGE, 25 of which were identified by MS/MS. Some proteins exhibited patterns of dispersion in the gel that are characteristic of post-translational modifications. Our findings suggest that the two sequencing platforms yielded similar results and that different omic approaches may be used to improve the understanding of gene expression. To adapt to low temperatures, E. antarcticum B7 expresses four of the six cold-shock proteins present in its genome. The cold-shock proteins were the most abundant in the bacterial proteome at 0°C. Some of the differentially expressed genes are required to preserve transcription and translation, while others encode proteins that contribute to the maintenance of the intracellular environment and appropriate protein folding. The results denote the complexity intrinsic to the adaptation of psychrotrophic organisms to cold environments and are based on two omic approaches. They also unveil the lifestyle of a bacterial species isolated in Antarctica.


Serum and urinary enzyme activity after renal infarction.
Recently it has been observed that the activity of certain enzymes in serum and urine may be increased after renal infarction. Although aortography or selective renal angiography should be the diagnostic corner-stone on which one would proceed to embolectomy, it is possible that enzyme assays may serve as laboratory aids to suggest or confirm the diagnosis. This paper reviews the few existing clinical and experimental studies and reports on two patients who had a total of three episodes of renal infarction. Serial determinations after one episode showed increased activity of serum oxaloacetic glutamic transaminase (SGOT) and of lactic acid dehydrogenase (LDH) and alkaline phosphatase in the serum and urine; some elevated serum LDH and SGOT values were recorded after the other two infarctions. The time of onset and duration of these increases are discussed, and the possible difficulty in differentiating renal from myocardial infarction is illustrated.


Don't forget the little brain: A framework for incorporating the cerebellum into the understanding of cognitive aging.
With the rapidly growing population of older adults, an improved understanding of brain and cognitive aging is critical, given the impacts on health, independence, and quality of life. To this point, we have a well-developed literature on the cortical contributions to cognition in advanced age. However, while this work has been foundational for our understanding of brain and behavior in older adults, subcortical contributions, particularly those from the cerebellum, have not been integrated into these models and frameworks. Incorporating the cerebellum into models of cognitive aging is an important step for moving the field forward. There has also been recent interest in this structure in Alzheimer's dementia, indicating that such work may be beneficial to our understanding of neurodegenerative disease. Here, I provide an updated overview of the cerebellum in advanced age and propose that it serves as a critical source of scaffolding or reserve for cortical function. Age-related impacts on cerebellar function further impact cortical processing, perhaps resulting in many of the activation patterns commonly seen in aging.