C-reactive protein impairs immune response of CD8+ T cells via FcγRIIb-p38MAPK-ROS axis in multiple myeloma.
C-reactive protein (CRP) is a prototypical acute phase protein in humans with the function of regulating immune cells. Serum CRP levels are elevated in multiple myeloma (MM), associated with MM cell proliferation and bone destruction. However, its direct effects on T lymphocytes in MM have not been elucidated. Public data sets were used to explore the correlation of CRP levels with immune cell infiltration and cytotoxicity score of CD8+ T cells in MM. In vitro, repeated freeze-thaw myeloma cell lines were taken as tumor antigens to load dendritic cells (DCs) derived from HLA-A*0201-positive healthy donors. MM-specific cytotoxic T cells (MM-CTL) were obtained from T lymphocytes of the corresponding donors pulsed with these DCs. B-cell maturation antigen (BCMA)-targeted chimeric antigen receptor (CAR)-T cells were manipulated by transfecting with lentivirus encoding an anti-BCMA single-chain variable fragment. Then T cells from healthy controls, MM-CTLs and BCMA CAR-T cells were exposed to CRP and analyzed for cell proliferation, cytotoxicity, immunophenotypes. CRP binding capacity to T cells before and after Fc gamma receptors IIb (FcγRIIb) blockage, p38 mitogen-activated protein kinase (MAPK) pathway and the downstream molecules were also detected. In vivo, both normal C57BL/6J mice and the Vk*MYC myeloma mouse models were applied to confirm the impact of CRP on T cells. CRP levels were negatively correlated with cell-infiltration and cytotoxicity score of CD8+ T cells in MM. In vitro experiments showed that CRP inhibited T-cell proliferation in a dose-dependent manner, impaired the cytotoxic activity and upregulated expression of senescent markers in CD8+ T cells. In vivo results validated the suppressive role of CRP in CD8+ T cells. CRP could bind to CD8+ T cells, mainly to the naïve T subset, while the binding was dramatically decreased by FcγRIIb blockage. Furthermore, CRP resulted in increased phosphorylation of p38 MAPK, elevated levels of reactive oxygen species and oxidized glutathione in CD8+ T cells. We found that CRP impaired immune response of CD8+ T cells via FcγRIIb-p38MAPK-ROS signaling pathway. The study casted new insights into the role of CRP in anti-myeloma immunity, providing implications for future immunotherapy in MM.


Knowledge, attitude and practices study on reproductive health among secondary school students in Bolgatanga, upper east region, Ghana.
This study was conducted within the secondary school student population of the Bolgatanga community, in Northern Ghana, to learn about knowledge, attitude and practices of reproductive health of this adolescent student population. Both quantitative and qualitative data were collected on adolescence perception, STIs and HIV/AIDS, family planning, male-female relationship, and vulnerability to sexual violence. The data collected show a concerning low familiarity of the student population with family planning methods and HIV/AIDS transmission, which, combined with minimal contraceptive use, pose them at high risk for unwanted pregnancies and sexual infections transmission. We argue that poor infrastructures and low accessibility of these rural areas in Northern Ghana may have led to uneven distribution of reproductive health educational programs in the country, urging more programs and interventions aimed particularly at these high risk groups.


[Study of the transmissibility of multiple drug resistance and the capacity to ferment lactose in a clinical strain of Kl. pneumoniae].
The donor properties of K. pneumoniae PI 220 with multiple drug resistance were studied. It was shown that the above strain carried 2 plasmids, i.e. R-plasmid pPI 220 controling resistance to ampicillin, streptomycin, tetracycline, chloramphenicol, kanamycin and sulphanylamides and plasmid pPI 221 controlling lactose fermentation. Both plasmids can be transfered on conjugation to strain E. coli P678 at a temperture of 28 degrees C at a rate of 10(-5) for pPI 220 and 10(-4) for pPI 221. The drug resistance controlled by pPI 221 was transfered mainly in a "blocks" simultaneously to 6 drugs. Deletion of plasmid pPI 220 was observed rarely. The donor properties of the strain were defined by the conjugative plasmid pPI 220 controlling the self-transfer and mobilization of plasmid pPI 221 incapable of the self-transfer. E. coli P678 (pPI 220) (PPI 221) acquired the donor properties and transfered both plasmids to E. coli J62 on crossing simultaneously at a rate of 10(-2), as well as to S. typhimurium LT2 and P. rettgeri at a rate of 10(-5). In all the recipient strains studied the transfered plasmids were unstable and segregated also simultaneously at a rate being the highest for P. retgari PI 230. The clones with stable preservation of the plasmids could be obtained by selection.


Ultraviolet fluorescence spectra of fingerprints.
We have studied inherent fluorescence spectra and imaging of fingerprints in the deep ultraviolet (UV) region with a nanosecond-pulsed Nd-YAG laser system that consists of a tunable laser, a cooled CCD camera, and a grating spectrometer. In this paper, we have studied UV fluorescence spectra of fingerprints under 266-nm illumination. Fluorescence spectra of fingerprints have two main peaks, around 330 nm (peak A) and 440 nm (peak B). At first, when a fingerprint has just been pressed, peak A is dominant. However, its intensity reduces as the total illumination time increases. On the other hand, peak B is weak at first. It appears after enough 266-nm illumination and its intensity increases as time elapses. After 3 h of illumination, peak A almost diminishes and peak B becomes dominant. By leaving the fingerprint under a fluorescent lamp in a room without laser illumination, peak A can be restored partly, while the intensity of peak B still increases. Time-resolved fluorescence spectra were also measured for these two peaks. The lifetime of each peak is 2.0 nsec (peak A) and 6.2 nsec (peak B) on average. Both peaks seem to consist of several components with different lifetimes. In the case of peak A, the 330-nm peak decays fast and a new component at 360 nm becomes dominant when the delay time exceeds 20 nsec. In the case of peak B, unlike peak A, no clear peak separation is observed, but the peak position seems to move from 440 to 460 nm when the delay time becomes larger.


Development of a radioimmunoassay for the fibrinogen-derived peptide B beta 1-42.
The peptide B beta 1-42 is the initial cleavage product of plasmin-mediated proteolysis of the NH2-terminal region of fibrinogen or fibrin I, while beta 15-42 is the major fragment released by plasmin degradation of fibrin II. Numerous studies have described the measurement of plasma B beta 1-42 levels as an index of plasmin activity. Previous assays were indirect and included quantitation of thrombin-increasable fibrinopeptide B immunoreactivity (TIFPB) or measurement of beta 15-42 with an antiserum (132) which cross-reacted with B beta 1-42. We report on a new antiserum (R142) directed against B beta 1-42 which does not cross-react with beta 15-42 or fibrinopeptide B. Employing this antiserum, a specific assay for B beta 1-42 was developed. This assay was used to measure plasmin-mediated B beta 1-42 release from fibrinogen and its subsequent proteolysis by thrombin. The selectivity constant (Kcat/Km) for thrombin cleavage of the B beta 14-15 bond of B beta 1-42 was 10(5)-fold less than that for proteolysis of the same bond in the intact fibrinogen molecule, thus explaining the stability of this peptide in the presence of thrombin activity in the blood. Similarly, the selectivity constant for plasmin cleavage of the B beta 21-22 bond of B beta 1-42 was 140-fold less than that for proteolysis of the B beta 42-43 bond of fibrinogen, indicating that secondary plasmin attack of the B beta 1-42 molecule is not physiologically important. The specific B beta 1-42 assay provided excellent recovery of peptide added to blood or plasma. Comparison of B beta 1-42 levels with TIFPB values in 37 patient samples yielded good correlation over a wide range of levels (r2 = 0.91). The median plasma B beta 1-42 level in 15 normal individuals was 1.2 pmol/mL. This is similar to the previously reported normal range for TIFPB (1 to 4 pmol/mL) but is higher than the normal level of 0.4 pmol/mL reported with the assay employing antiserum 132. This discrepancy reflects rapid removal of Arg (B beta 42) by plasma carboxypeptidase activity resulting in 50% loss of B beta 1-42 immunoreactivity with antiserum 132 but no loss with R142. To circumvent this problem, we have developed a beta 15-42 antiserum (R154) which, like antiserum 132, cross-reacts with B beta 1-42. However, B beta 1-42/beta 15-42 immunoreactivity with R154 is stable in the presence of carboxypeptidase activity.(ABSTRACT TRUNCATED AT 400 WORDS)


Spinal opioid delta antinociception in the mouse: mediation by a 5'-NTII-sensitive delta receptor subtype.
Previous studies from our laboratory have indicated that i.c.v. pretreatment of mice with the novel, selective opioid delta receptor antagonists, [D-Ala2,Leu5,Cys6]enkephalin (DALCE) and naltrindole-5'-isothiocyanate (5'-NTII), differentially antagonized the direct antinociceptive effects of [D-Pen2,D-Pen5]enkephalin (DPDPE) and [D-Ala2]deltorphin II (DELT). These findings, and others, suggested the existence of subtypes of opioid delta receptors which could be classified as activated by DPDPE and DALCE sensitive (delta 1 receptor), or selectively activated by DELT and 5'-NTII sensitive (delta 2 receptor). The present study has extended these observations to the characterization of delta-mediated antinociception effects of DPDPE and DELT after i.t. administration in mice using pretreatment with DALCE and 5'-NTII in order to selectively antagonize the delta subtypes. Additionally, the acute antinociceptive actions of DALCE itself were studied to ensure activity of this compound at the spinal level. The respective antinociceptive A50 value (95% CL) for i.t. DPDPE, DELT and DALCE were 19.0 (12.9-28.1), 19.3 (16.1-23.1) and 2.0 (1.4-3.0) nmol. The delta antagonist, N,N-diallyl-Try-Aib-Aib-Phe-Leu-OH (ICI 174,864) (where Aib is alpha-aminoisobutyric acid) blocked the antinociceptive effects of DPDPE and DELT, but not those of i.t. morphine or [D-Ala2,NMPhe4,Gly-ol5]enkephalin (DAMGO), indicating that the observed antinociceptive effects of DPDPE and DELT were delta mediated. Pretreatment 24 hr before testing with graded doses of i.t. 5'-NTII blocked the i.t. antinociceptive effects of DPDPE and DELT, although at least a 10-fold higher dose of 5'-NTII was needed to produce equivalent antagonism of DPDPE.(ABSTRACT TRUNCATED AT 250 WORDS)


Pathogen surveillance in the informal settlement, Kibera, Kenya, using a metagenomics approach.
Worldwide, the number of emerging and re-emerging infectious diseases is increasing, highlighting the importance of global disease pathogen surveillance. Traditional population-based methods may fail to capture important events, particularly in settings with limited access to health care, such as urban informal settlements. In such environments, a mixture of surface water runoff and human feces containing pathogenic microorganisms could be used as a surveillance surrogate. We conducted a temporal metagenomic analysis of urban sewage from Kibera, an urban informal settlement in Nairobi, Kenya, to detect and quantify bacterial and associated antimicrobial resistance (AMR) determinants, viral and parasitic pathogens. Data were examined in conjunction with data from ongoing clinical infectious disease surveillance. A large variation of read abundances related to bacteria, viruses, and parasites of medical importance, as well as bacterial associated antimicrobial resistance genes over time were detected. Significant increased abundances were observed for a number of bacterial pathogens coinciding with higher abundances of AMR genes. Vibrio cholerae as well as rotavirus A, among other virus peaked in several weeks during the study period whereas Cryptosporidium spp. and Giardia spp, varied more over time. The metagenomic surveillance approach for monitoring circulating pathogens in sewage was able to detect putative pathogen and resistance loads in an urban informal settlement. Thus, valuable if generated in real time to serve as a comprehensive infectious disease agent surveillance system with the potential to guide disease prevention and treatment. The approach may lead to a paradigm shift in conducting real-time global genomics-based surveillance in settings with limited access to health care.


Transferring the occlusal and esthetic anatomy of the provisional to the final restoration in full-arch oral rehabilitations.
Provisional restorations are an excellent diagnostic instrument, especially in full-arch oral rehabilitations where no normal occluding contacts remain. Esthetics, phonetics, function, parafunction, and dysfunction after evaluation and acceptance through clinical trial with the provisional restorations should be accurately transferred to the final restorations to ensure the same clinical success. This article presents a sequence of clinical procedures that enables a methodical and controlled transfer of the morphology of the provisional restorations to the final ones in full-arch oral rehabilitations.


Insulin sensitivity and liver insulin receptor structure in ducks from two genera.
Insulin sensitivity and liver insulin receptor structure were studied in 5-wk-old ducks from two genera (Muscovy and Pekin). In the fasting state, both duck types were equally resistant to exogenous insulin compared with chicken. Despite the low potency of duck insulin, the number of insulin receptors was lower in Muscovy duck and similar in Pekin duck and chicken liver membranes. After 125I-insulin cross-linking, the size of the alpha-subunit of the receptors from the three species was 135,000. Wheat germ agglutinin-purified receptors from the three species were contaminated by an active and unusual adenosinetriphosphatase (ATPase) contaminant (highest activity in Muscovy duck). Sequential purification of solubilized receptor from both duck types on lentil and then wheat germ agglutinin lectins led to a fraction of receptors very poor in ATPase activity that exhibited a beta-subunit size (95,000) and tyrosine kinase activity similar to those of ATPase-free chicken insulin receptors. Therefore the ducks from the two genera exhibit an alpha-beta-structure for liver insulin receptors and a clear difference in the number of liver insulin receptors. Their sensitivity to insulin is, however, similarly decreased compared with chicken.


Effect of Chlorella vulgaris extracts on murine cytomegalovirus infections.
The host-mediated antiviral effect of Chlorella vulgaris extracts (CVE) was evaluated in ICR mice against murine cytomegalovirus (MCMV) infection. Mice treated with 10 mg of CVE on days 3 and 1 before virus challenge survived lethal infection. The protective effect of CVE was shown by a decrease in the infectious viruses replicated in the target organs of CVE-treated ICR mice. CVE also protected mice from histopathological damage to the target organs due to MCMV infection. Both the serum interferon (IFN) level and 2'5'-oligoadenylate (2-5A) synthetase activity were elevated in CVE-treated and MCMV-infected mice, and were higher than those in the control mice. The natural killer (NK) activity of spleen cells, which is otherwise deteriorated by lethal MCMV infection, was remarkably augmented in CVE-treated mice. Neither virocidal nor virostatic activity of CVE on MCMV was seen in vitro. Thus, CVE-induced resistance against MCMV seems to be host-mediated. The protective effect correlated with the augmentation of NK activity of the spleen cells and serum IFN activity, which inhibited replication of MCMV in the target organs and subsequently saved the mice from death.


Inhibition Effectiveness of Laser-Cleaned Nanostructured Aluminum Alloys to Sulfate-reducing Bacteria Based on Superwetting and Ultraslippery Surfaces.
This paper is a continued study on laser cleaning removal of marine microbiofouling from Al alloy surfaces. According to our previous study, it is noted that the antifouling functions of the generated laser-cleaned metallic surfaces must be highlighted. In this work, the inhibition effectiveness of the laser-cleaned Al alloy surfaces was evaluated using a type of vital marine microorganism, sulfate-reducing bacteria (SRB) Desulfovibrio desulfuricans subsp. desulfuricans, in a dynamic bacterial solution. Before the immersion tests, the laser-cleaned surfaces with nanostructures were chemically processed into superhydrophilic, superhydrophobic, and ultraslippery surfaces. SRB attachment behaviors as well as inhibition mechanisms of the three surfaces to the SRB settlement were characterized and revealed. The SRB adhering to the above surfaces presented three different morphologies, i.e., broken, dented, and plump cells. Superhydrophilic surfaces unexpectedly showed a not inferior antibacterial ability. A piercing effect of the nanostructures caused nontoxic mechanical damage to the cell membranes. The antiadhesion property of superhydrophobic solid-air hybrid surfaces was unreliable due to the loss of air bubbles. The morphology of the last surviving SRB cells left on the ultraslippery surfaces was basically plump. The stable repellent function of the surfaces was responsible for the vigorous prevention of the adhesion of the SRB. The research results offer an insight into the antibacterial/antiadhesion properties of the laser-cleaned surfaces and a practical value for the periodic service of marine high-end equipment.


Role for Surgical Resection in the Multidisciplinary Treatment of Stage IIIB Non-Small Cell Lung Cancer.
The role of multimodality therapy in stage IIIB non-small cell lung cancer (NSCLC) remains inadequately studied. Although chemoradiation is currently the mainstay of treatment, randomized trials evaluating surgical intervention are lacking, and resection is offered selectively. Data from patients with clinical stage IIIB NSCLC (T4N2 or any N3) undergoing definitive multimodality therapy were obtained from the National Cancer Database (NCDB). Multivariable Cox regression models were fitted to evaluate variables influencing overall survival (OS). From 1998 to 2010, 7,459 patients with clinical stage IIIB NSCLC were treated with definitive chemoradiation (CR group), whereas 1,714 patients underwent chemotherapy, radiation, and surgical intervention in any sequence (CRS group). CRS patients were more likely to be younger and white and have slightly smaller tumors (all p < 0.01). There was no difference in Charlson Comorbidity Index (CCI) between the groups (p = 0.5). In the CRS group, 79% of patients received neoadjuvant therapy. Thirty-day surgical mortality was 3%. Factors associated with improved OS in multivariate analysis included younger age, female sex, decreased CCI, smaller tumor size, and surgical resection (hazard ratio [HR], 0.57; 95% confidence interval [CI], 0.52-0.63). Among patients treated with surgical intervention, incomplete resection was associated with decreased OS (HR, 1.52; 95% CI, 1.20-1.92). Median OS was longer in the CRS group (25.9 months versus 16.3 months; p < 0.001). Propensity matched analysis on 631 patient pairs treated with CRS versus CR confirmed these findings (median OS, 28.9 versus 17.2 months; p < 0.001). Surgical resection as a part of multimodality therapy may be associated with improved OS in highly selected patients with stage IIIB NSCLC. Multidisciplinary evaluation of these patients is critical.


Lighting up yeast cell factories by transcription factor-based biosensors.
Our ability to rewire cellular metabolism for the sustainable production of chemicals, fuels and therapeutics based on microbial cell factories has advanced rapidly during the last two decades. Especially the speed and precision by which microbial genomes can be engineered now allow for more advanced designs to be implemented and tested. However, compared to the methods developed for engineering cell factories, the methods developed for testing the performance of newly engineered cell factories in high throughput are lagging far behind, which consequently impacts the overall biomanufacturing process. For this purpose, there is a need to develop new techniques for screening and selection of best-performing cell factory designs in multiplex. Here we review the current status of the sourcing, design and engineering of biosensors derived from allosterically regulated transcription factors applied to the biotechnology work-horse budding yeast Saccharomyces cerevisiae. We conclude by providing a perspective on the most important challenges and opportunities lying ahead in order to harness the full potential of biosensor development for increasing both the throughput of cell factory development and robustness of overall bioprocesses.


High-resolution FRET microscopy of cholera toxin B-subunit and GPI-anchored proteins in cell plasma membranes.
"Lipid rafts" enriched in glycosphingolipids (GSL), GPI-anchored proteins, and cholesterol have been proposed as functional microdomains in cell membranes. However, evidence supporting their existence has been indirect and controversial. In the past year, two studies used fluorescence resonance energy transfer (FRET) microscopy to probe for the presence of lipid rafts; rafts here would be defined as membrane domains containing clustered GPI-anchored proteins at the cell surface. The results of these studies, each based on a single protein, gave conflicting views of rafts. To address the source of this discrepancy, we have now used FRET to study three different GPI-anchored proteins and a GSL endogenous to several different cell types. FRET was detected between molecules of the GSL GM1 labeled with cholera toxin B-subunit and between antibody-labeled GPI-anchored proteins, showing these raft markers are in submicrometer proximity in the plasma membrane. However, in most cases FRET correlated with the surface density of the lipid raft marker, a result inconsistent with significant clustering in microdomains. We conclude that in the plasma membrane, lipid rafts either exist only as transiently stabilized structures or, if stable, comprise at most a minor fraction of the cell surface.


Lack of chronic oral toxicity of chemopreventive bovine lactoferrin in F344/DuCrj rats.
Studies were undertaken to determine whether bovine lactoferrin (bLF) and related compounds, shown to prevent carcinogenesis in the colon and other organs in rats, have any toxic effects in long-term feeding studies. In experiment I, male F344/DuCrj rats received a basal diet containing 0.2% bLF for 40 weeks. No adverse findings were noted, furthermore, serum triglyceride level was significantly decreased to 72% of the control level, suggesting preventive effects against the metabolic syndrome. In experiment II, male and female F344/DuCrj rats were fed a basal diet containing 0.02, 0.2, 2.0 and 5.0% bLF, 2.0% bLF hydrolysate (bLF-H) or 0.1% lactoferricin (LFcin), a peptide derived from bLF, for 60 weeks in males and 65 weeks in females. No toxicological effects, including carcinogenicity, were evident in either sex. The results of the studies provide subjective support for safety of clinical studies of bLF for supplement use.


Borrelia burgdorferi antibodies in outdoor and indoor workers in south-west Sweden.
Two hundred and fifty-three farmers and forest workers and 249 clerks from south-west Sweden were recruited to a cross-sectional seroprevalence study to find out if individuals working outdoors are more prone to acquire Borrelia burgdorferi infection than indoor workers and to find undiagnosed cases of Lyme borreliosis. The participants answered a questionnaire and blood specimens were collected to estimate the prevalence of antibodies to B. burgdorferi in each group. Sera were analysed with an enzyme-linked immunoassay technique to determine IgG antibodies to B. burgdorferi flagellum. The prevalence of B. burgdorferi antibodies was 7.6% in the farmers and forest workers vs. 5.3% in the clerks (adjusted odds ratio [age, sex] = 1.2 [95% confidence interval = 0.5-2.8]). One case of Lyme borreliosis was diagnosed. The positive predictive value of the antibody test was estimated to be 3% in the studied populations. B. burgdorferi infection is of low endemicity in south-west Sweden and is probably not an occupational risk among outdoor workers. Undiagnosed cases of Lyme borreliosis are uncommon. The test used is not acceptable for screening purposes.


Usefulness of faecal calprotectin measurement in children with various types of inflammatory bowel disease.
The aim of the study was to assess the usefulness of the FC measurement in children with various types of IBD and relation to the disease activity. 91 patients (49 boys: 53.85% and 42 girls: 46.15%, mean age: 13.38 years, range 6-18 years) were included in the analysis. Patients were divided into the groups: B1-24 children with CD, B2-16 patients with UC, and a group comprising 31 children with other types of colitis; the control group (K) comprised 20 healthy children. FC was assayed by ELISA method, using Phical test (Calpro). The mean faecal calprotectin concentrations were higher in children with CD and UC as compared to healthy controls, patients with eosinophilic, lymphocytic, and nonspecific colitis. A positive correlation was observed between FC concentrations and the disease activity (the PCDAI scale, the Truelove-Witts Scale, and the endoscopic Rachmilewitz Index). It seems that the FC concentrations can be a useful, safe, and noninvasive test in children suspected for IBD, since FC concentration is higher in children with CD and UC than in patients with other inflammatory diseases.


Radioimmunoassay of calcitonin in plasma, normal thyroid, and medullary thyroid carcinoma of the rat.
We have developed a sensitive immunoassay for murine calcitonin which can measure the basal concentration of the hormone in peripheral plasma of rats as well as the calcitonin, extracted from rat thyroid tissue. Human calcitonin was used for tracer, standard, and antibody production. Assay methods were devised to minimize artifacts which have been shown to spuriously influence immunoassay performance. The basal plasma concentration of calcitonin in rats was 6 to 75 pg. per milliliter. Infusions of calcium, pentagastrin, and PGE2 produced a 2- to 7-fold increase in plasma calcitonin. Elevated concentrations of calcitonin were demonstrated in tumor specimens obtained from the second and fourth generations of a transplanted rat medullary thyroid carcinoma. Our results demonstrate that calcitonin circulates in the peripheral plasma of rats in the basal state and that hormone concentrations change during functional tests of secretion. This assay will permit studies of calcitonin secretion in physiological and pathophysiological states in the rat.


Central angiotensin II stimulates cutaneous water intake behavior via an angiotensin II type-1 receptor pathway in the Japanese tree frog Hyla japonica.
Angiotensin II (Ang II) stimulates oral water intake by causing thirst in all terrestrial vertebrates except anurans. Anuran amphibians do not drink orally but absorb water osmotically through ventral skin. In this study, we examined the role of Ang II on the regulation of water-absorption behavior in the Japanese tree frog (Hyla japonica). In fully hydrated frogs, intracerebroventricular (ICV) and intralymphatic sac (ILS) injection of Ang II significantly extended the residence time of water in a dose-dependent manner. Ang II-dependent water uptake was inhibited by ICV pretreatment with an angiotensin II type-1 (AT(1)) receptor antagonist but not a type-2 (AT(2)) receptor antagonist. These results suggest that Ang II stimulates water-absorption behavior in the tree frog via an AT(1)-like but not AT(2)-like receptor. We then cloned and characterized cDNA of the tree frog AT(1) receptor from the brain. The tree frog AT(1) receptor cDNA encodes a 361 amino acid residue protein, which is 87% identical to the toad (Bufo marinus) AT(1) receptor and exhibits the functional characteristics of an Ang II receptor. AT(1) receptor mRNAs were found to be present in a number of tissues including brain (especially in the diencephalon), lung, large intestine, kidney and ventral pelvic skin. When tree frogs were exposed to dehydrating conditions, AT(1) receptor mRNA significantly increased in the diencephalon and the rhombencephalon. These data suggest that central Ang II may control water intake behavior via an AT(1) receptor on the diencephalon and rhombencephalon in anuran amphibians and may have implications for water consumption in vertebrates.


Linear theory of ionization cooling in 6D phase space.
A linear theory is developed for ionization cooling of muon beams in periodic channels that can provide cooling of the transverse emittances and also of the longitudinal emittance via emittance exchange. The channels incorporate solenoids and quadrupoles for transverse focusing, dipoles to generate dispersion, wedged absorbers for ionization, and rf cavities for acceleration. The beam evolution near equilibrium is described by coupled first-order differential equations for five generalized emittances with two excitation sources. The results should be useful for understanding the cooling process and for designing cooling channels of future muon colliders and neutrino factories.


Combined anterior cruciate-ligament reconstruction using semitendinosus tendon and iliotibial tract.
We are reporting the results of a reconstructive procedure designed to decrease anterior tibial subluxation due to disruption of the anterior cruciate ligament. The operation combines both intra-articular and extra-articular methods. The semitendinosus tendon and the iliotibial tract are both routed from opposite directions over the top of the lateral femoral condyle and through the same oblique drill-hole in the proximal part of the tibia: the semitendinosus tendon is passed up through the tibial drill-hole, across the knee joint, over the top of the lateral femoral condyle, and deep to the fibular collateral ligament, and the iliotibial tract is passed deep to the fibular collateral ligament, over the top of the lateral femoral condyle, across the knee joint, and down through the drill-hole. Both grafts are simultaneously pulled tight while the semitendinosus tendon is sutured to the iliotibial tract laterally and the iliotibial tract is sutured to the semitendinosus tendon medially below the drill-hole. The posteromedial and lateral parts of the capsule are advanced to tighten the secondary restraints. One hundred of the first 106 consecutive patients with chronic instability who had this procedure were evaluated using subjective and objective criteria at three to seven and one-half years after surgery. The positive anterior-drawer sign tested at 25 degrees of flexion was eliminated or reduced to 1+ in eighty knees, and the positive pivot shift was reduced to zero or 1+ in ninety-one knees. The objective assessment of isokinetic muscle performance and passive tibial rotation showed significant improvements in strength and normalization of tibial rotation.


[Unusual retinal anomaly: the double papilla].
It is not clear whether doubling of the papilla occurs in connection with congenital ocular malformations or whether there is a double anlage of a real papilla. A double optic nerve close to the globe has never been observed. A 87-year old woman had high-grade myopia and amblyopia of her right eye together with an esotropia from early childhood. The right eye had a cataract. Some time ago she experienced a bilateral incomplete infarction of the territory of the posterior cerebral artery. The visual acuity of her right eye was 1/50. The Goldmann visual field showed a concentric constriction with the V/4 test mark. The right fundus revealed a double configuration of a papilla, in addition to widespread myopic changes due to extension of the retina and choroid at the posterior eye pole. The two papillae were located one upon another and the slightly smaller pseudopapilla was found temporal beneath to the true papilla. The papillae had central vessels, well supplied with blood, and with distributions into temporal and nasal tributaries. The big vessels of both papillae were not connected. The MRI revealed only one optic nerve of normal appearance. The findings in our patient demonstrate that a double papilla occurs without a double optic nerve. The smaller papilla occurred in an area of widespread myopic changes of the retina and choroid that we assume the appearance of a pseudopapilla. The site beneath the true papilla in an colobomatous area points to a developmental failure due to an incomplete closure of the optic cleft of the embryo. A secondary ingrow of vessels could have developed into the pseudopapilla during the embryonal period.


Clinical Characteristics of 90 Macrodactyly Cases.
Macrodactyly is a rare, nonhereditary congenital deformity. Digital enlargement in macrodactyly involves all tissue types and presents alone or as part of a congenital deformity syndromes. Macrodactyly treatment largely depends on surgeons' experience and knowledge. Because there is a paucity of large cohort studies of macrodactyly in the literature, our goal was to retrospectively analyze macrodactyly cases in order to define a better system for diagnosis, classification, and prognosis. Medical records of 90 Chinese macrodactyly patients, including demographic characteristics, clinical presentations, anatomical distributions, x-rays, pathological findings, and treatments, were reviewed. Genetic analyses of 12 patients were also reviewed. Disease incidence was similar across sex and geographical regions. Multiple-digit involvement was 2.6 times more frequent than single-digit involvement. The index finger, middle finger, and thumb were most commonly involved. Two digits were affected more often than 3, with the affected digits adjacent in most cases. The affected digit was in the median nerve innervation distribution in 79% of cases and was accompanied by enlargement and fat infiltration of the median nerve. Seven cases had syndactyly. Ten of the 12 cases subjected to PIK3CA mutation analysis were positive. Macrodactyly represents a heterogeneous group of conditions, without significant sex or geographical predilection, which is usually present at birth. A high PIK3CA mutation-positive rate in affected tissues suggests a similar cellular mechanism for overgrowth in patients with various clinical presentations. Prognostic IV.


Lipoic acid attenuates hypertension and improves insulin sensitivity, kallikrein activity and nitrite levels in high fructose-fed rats.
Chronic feeding of fructose to normal rats causes impaired glucose tolerance, loss of tissue sensitivity to insulin, hyperinsulinemia and hypertension. alpha-Lipoic acid (LA), a co-enzyme known for its potent antioxidant effects, stimulates insulin-mediated glucose uptake in clinical and experimental diabetes. The purpose of this study was to examine whether LA can mitigate fructose-induced insulin resistance and associated abnormalities. Male Wistar rats of body weights 150-170 g were divided into 4 groups containing 12 rats each. Control rats received a control diet containing starch and water ad libitum. Fructose rats received a fructose-enriched diet (>60% of total calories). Fructose + LA rats received a fructose diet and LA (35 mg/kg b.w.) intraperitoneally. Control + LA rats received a normal diet and LA (35 mg/kg b.w.) intraperitoneally. After the treatment period of 20 days, blood pressure (BP) was measured. Oral glucose-tolerance test, insulin-sensitivity index, urea and creatinine clearance tests, and plasma and urinary sodium and potassium levels were analysed. Kallikrein activity and nitrite content were assayed. Additionally, the activities of RBC-membrane Na(+)/K(+) ATPase and Ca(2+) ATPase enzymes were assayed. Fructose rats showed increased BP, decreased glucose tolerance, decreased insulin sensitivity and altered sodium and potassium levels and renal clearance. LA supplementation mitigated these alterations. The increase in BP was attenuated and the levels of biochemical parameters were brought close to normal. The BP-lowering effect of LA in fructose rats may be related to improvement in insulin sensitivity.


Bubble-based acoustic micropropulsors: active surfaces and mixers.
Acoustic micropropulsors present great potential for microfluidic applications. The propulsion is based on encapsulated 20 μm bubbles excited by a contacless ultrasonic transducer. The vibrating bubbles then generate a powerful streaming flow, with speeds 1-100 mm s-1 in water, through the action of viscous stresses. In this paper we introduce a full toolbox of micropropulsors using a versatile three-dimensional (3D) microfabrication setup. Doublets and triplets of propulsors are introduced, and the flows they generate are predicted by a theoretical hydrodynamic model. We then introduce whole surfaces covered with propulsors, which we term active surfaces. These surfaces are excited by a single ultrasonic wave, can generate collective flows and may be harnessed for mixing purposes. Several patterns of propulsors are tested, and the flows produced by the two most efficient mixers are predicted by a simple theoretical model based on flow singularities. In particular, the vortices generated by the most efficient pattern, an L-shaped mixer, are analysed in detail.


Higher set point plasma viral load and more-severe acute HIV type 1 (HIV-1) illness predict mortality among high-risk HIV-1-infected African women.
There is limited information on the natural history of human immunodeficiency virus type 1 (HIV-1) infection in Africa, especially from individuals with well-defined dates of infection. We used data from a prospective cohort study of female sex workers in Mombasa, Kenya, who were followed up monthly from before the date of HIV-1 infection. Antiretroviral-naive women who had a well-defined date of HIV-1 infection were included in this analysis. The effects of set point plasma viral load (measured 4-24 months after infection), early CD4+ cell count, and symptoms of acute HIV-1 infection on mortality were assessed using Cox proportional hazards analysis. Among 218 women, the median duration of follow-up after HIV-1 infection was 4.6 years. Forty women died, and at 8.7 years (the time of the last death), the cumulative survival rate was 51% by Kaplan-Meier analysis. Higher set point viral load, lower early CD4+ cell count, and more-symptomatic acute HIV-1 illness each predicted death. In multivariate analysis, set point viral load (hazard ratio [HR], 2.28 per 1 log10 copies/mL increase; P=.001) and acute HIV-1 illness (HR, 1.14 per each additional symptom; P=.05) were independently associated with higher mortality. Among this group of African women, the survival rate was similar to that for HIV-1-infected individuals in industrialized nations before the introduction of combination antiretroviral therapy. Higher set point viral load and more-severe acute HIV-1 illness predicted faster progression to death. Early identification of individuals at risk for rapid disease progression may allow closer clinical monitoring, including timely initiation of antiretroviral treatment.


The self-assessment Global Quality of Life scale: Reliability and construct validity.
Abstract Objective. The aim was to establish psychometric properties of the Global Quality of Life Scale (GQL) for people with severe mental illness. Methods. GQL is a stand-alone visual analogue scale included in "The Quality Star", a minimal platform for clinical follow-up and efficiency documentation of mental health services in eight dimensions widely used in Sweden. Validating instruments included MANSA, Inventory of Problem and Solutions, Consumer Satisfaction Rating Scale, Perceived Global Distress, health screening using UKU-Side Effect Rating Scale, GAF, and Perceived Global Burden (for next of kin). Test-retest reliability of the GQL was examined between ratings at quarterly intervals during 1 year. Results. There were three main results: test-retest reliability at quarterly intervals was very satisfactory, concurrent validity with the initial item of life satisfaction scale of MANSA, "Life as a whole", was demonstrated and finally content validity was clarified by associations with a number of validating measures from several contexts in three studies. Conclusion. GQL have acceptable psychometric properties and is valid for serious mental ill persons. Its use as easy-to-use instrument for screening of perceived global quality of life was supported.


Bipolar Electrochemical Measurement of Enantiomeric Excess with Inherently Chiral Polymer Actuators.
Straightforward enantioselective analytical methods are very important for drug safety, considering that in certain cases one of the two enantiomers of a chiral molecule might be harmful for humans. In this work, we propose a simple system for the direct and easy read-out of the enantiomeric excess of 3,4-dihydroxyphenylalanine (DOPA) as a model analyte. A conducting oligomer, i.e. oligo-(3,3'-dibenzothiophene), bearing inherently chiral features, is electrogenerated on a polypyrrole film. The resulting freestanding hybrid material is used as a wireless enantioselective actuator in a bipolar electrochemical cell. Combining in a single setup two individual actuators with opposite chiral features allows a direct visual read-out of enantiomeric excess, as the bending amplitude of each of the two actuators is directly correlated with the concentration of the corresponding stereoisomer of the analyte. Optimization of the experimental parameters results in efficient bending, giving access to the percentage values of the enantiomeric excess in mixtures containing different ratios of the antipodes, thus opening the way to potential applications for chiral in situ analysis.


Effect of insulin on in vivo glucose utilization in individual tissues of anesthetized lactating rats.
Glucose utilization rate has been measured in skeletal muscles, white adipose tissue, and mammary gland of anesthetized nonlactating and lactating rats. During lactation, basal glucose utilization is decreased by 40% in periovarian white adipose tissue and by 65% in epitrochlearis and extensor digitorum longus but not in soleus muscle. This may be related to the lower blood glucose and plasma insulin concentrations observed during lactation. Basal glucose utilization rate in the mammary gland was, respectively, 18 +/- 2 and 350 +/- 50 micrograms/min in nonlactating and lactating rats. During the euglycemic hyperinsulinemic clamp, a physiological increment in plasma insulin concentration (231 +/- 18 in lactating vs. 306 +/- 24 microU/ml in nonlactating rats) induces a similar increase in glucose utilization rate in skeletal muscles (except soleus) and white adipose tissue in the two groups of rats. Furthermore this low increase in plasma insulin concentration does not alter mammary glucose utilization rate in nonlactating rats but induces the same increase (sevenfold over basal) as a maximal insulin concentration in lactating rats. These data show that the active mammary gland is the most insulin-sensitive tissue of the lactating rat that has been tested. The overall increase in insulin sensitivity and responsiveness that has been described in lactating rats can then mainly be attributed to the presence of the active mammary gland.


Bioprinting: A Strategy to Build Informative Models of Exposure and Disease.
Novel additive manufacturing techniques are revolutionizing fields of industry providing more dimensions to control and the versatility of fabricating multi-material products. Medical applications hold great promise to manufacture constructs of mixed biologically compatible materials together with functional cells and tissues. We reviewed technologies and promising developments nurturing innovation of physiologically relevant models to study safety of chemicals that are hard to reproduce in current models, or diseases for which there are no models available. Extrusion-, inkjet- and laser-assisted bioprinting are the most used techniques. Hydrogels as constituents of bioinks and biomaterial inks are the most versatile materials to recreate physiological and pathophysiological microenvironments. The highlighted bioprinted models were chosen because they guarantee post-printing cellular viability while maintaining desirable mechanical properties of their constitutive bioinks or biomaterial inks to ensure their printability. Bioprinting is being readily adopted to overcome ethical concerns of in vivo models and improve the automation, reproducibility, geometry stability of traditional in vitro models. The challenges for advancing the technological level readiness of bioprinting require overcoming heterogeneity, microstructural complexity, dynamism and integration with other models, to generate multi-organ platforms that can inform about biological responses to chemical exposure, disease development and efficacy of novel therapies.


Establishment of lysozyme gene RNA interference systemand its involvement in salinity tolerance in sea cucumber (Apostichopus japonicus).
The lysozyme gene was silenced using RNA interference (RNAi) to analyze the function of lysozyme in sea cucumber under salt stress. The interfering efficiency of four lysozyme RNAi oligos ranged from 0.55 to 0.70. From the four oligos, p-miR-L245 was used for further interfering experiments because it had the best silencing efficiency. Peristomial film injection of p-miR-L245 (10 μg) was used for further interfering experiments. The lowest gene expression, determined by RT-PCR assay, in muscle, coelomic fluid, and parapodium occurred 48 h after p-miR-L245 injection, while that of body wall and tube foot was 96 h and 24 h, respectively. Lysozyme activity in muscle and body wall was significantly lower than the controls. The lowest lysozyme activity in muscle, body wall and parapodium, was found at 48, 72, and 48 h, respectively, which was consistent with the transcription expression of lysozyme. The lowest point of lysozyme activity was at 96 h in coelomic fluid and tube foot, which was laid behind lysozyme expression in transcription level. The expression profile of the lysozyme transcription level and lysozyme activity in the body wall and tube foot increased at 12 h after p-miR-L245 injection before the interference effect appeared. NKA gene expression was expressed at a high level in the positive control (PC) and negative control (NC) groups at 12, 24, and 48 h, while NKA was expressed at low levels in the lysozyme RNAi injection group at 12 and 24 h. The level of NKA gene expression recovered to the level of the PC and NC group at 48, 72, and 96 h after the lysozyme RNAi injection. NKCC1 gene expression was high in the PC and NC groups at 96 h, while the NKCC1 was expressed at a low level 96 h after lysozyme RNAi injection. The results suggest that lysozyme decay involves NKA and NKCC1 gene expression under salinity 18 psμ. The K+ and Cl- concentration after lysozyme RNAi injection was lower than in the PC and NC group.


Determination of the number of events required for mammary carcinogenesis in the Sprague-Dawley female rat.
Female Sprague-Dawley rats were exposed to a single, graded dose of either of two highly effective mammary chemical carcinogens, 7,12-dimethylbenzanthracene (DMBA) or N-methylnitrosourea, in order to determine the number of mammary cancers per rat induced by a range of carcinogenic doses. These data were then used to separately construct dose-response curves characteristic for DMBA- and N-methylnitrosourea-induced mammary carcinogenesis. Analysis of these characteristic dose-response curves demonstrated that, following a single exposure to either DMBA or N-methylnitrosourea, the number of mammary cancers per rat increased not linearly but as the second power of dose of carcinogen used. These results are clearly incompatible with mammary carcinogenesis being a single step process in the female Sprague-Dawley rat. In direct contrast these results are entirely consistent with a malignant process requiring two transformation events. When female Sprague-Dawley animals are exposed multiple times to a suboptimal dose of DMBA, the number of mammary cancers induced per rat increases synergistically, not merely additively, as compared to a single dose exposure. Again this result is consistent only with mammary carcinogenesis requiring at least two transformation events.


[Dependence of the results of a genetic analysis of self-pollinating cereal species on the specificity of the mapping population].
The available in literature information as for results of genetic analysis of several self-pollinating cereal species involving the use of plant lines with alien genetic material as the object of analysis is summarized. The restrictions caused by specificity of the investigated material as for the possible explanation of the obtained results are emphasized. Several suggestions concerning the preliminary study of introgressive plant lines before their including into experimental programs on genetic analysis are made.


Arsenic Trioxide inhibits Activation of Hedgehog Pathway in Human Neuroblastoma Cell Line SK-N-BE(2) Independent of Itraconazole.
Neuroblastoma (NB) remains associated with a low overall survival rate over the long term. Abnormal activation of the Hedgehog (HH) signaling pathway can activate the transcription of various downstream target genes that promote NB. Both arsenic trioxide (ATO) and itraconazole (ITRA) can inhibit tumor growth. To determine whether ATO combined with ITRA can be used to treat NB with HH pathway activation, we examined the effects of ATO and ITRA monotherapy or combined inhibition of the HH pathway in NB. Analysis of CCK8 and flow cytometry showed cell inhibition and cell cycle, respectively. Real-time PCR analysis was conducted to assess the mRNA expression of HH pathway. We revealed that as concentrations of ATO and ITRA increased, the killing effects of both agents on SK-N-BE(2) cells became more apparent. During G2/M, the cell cycle was largely arrested by ATO alone and combined with ITRA, and in the G0/G1 phase by ITRA alone. In the HH pathway, ATO inhibited the transcription of the SHH, PTCH1, SMO and GLI2 genes, however, ITRA did not. Instead of showing synergistic effects in a combined mode, ITRA decreased ATO inhibitory effects. We showed that ATO is an important inhibitor of HH pathway but ITRA can weaken the inhibitory effect of ATO. This study provides an experimental evidence for the clinical use of ATO and ITRA in the treatment of NB with HH pathway activation in cytology.


Cyperus rotundus suppresses AGE formation and protein oxidation in a model of fructose-mediated protein glycoxidation.
Non-enzymatic glycation, as the chain reaction between reducing sugars and the free amino groups of proteins, has been shown to correlate with severity of diabetes and its complications. Cyperus rotundus (Cyperaceae) is used both as a food to promote health and as a drug to treat certain diseases. In this study, considering the antioxidative effects of C. rotundus, we examined whether C. rotundus also protects against protein oxidation and glycoxidation. The protein glycation inhibitory activity of hydroalcoholic extract of C. rotundus was evaluated in vitro using a model of fructose-mediated protein glycoxidation. The C. rotundus extract with glycation inhibitory activity also demonstrated antioxidant activity when a ferric reducing antioxidant power (FRAP) and Trolox equivalent antioxidant capacity (TEAC) assays as well as metal chelating activity were applied. Fructose (100mM) increased fluorescence intensity of glycated bovine serum albumin (BSA) in terms of total AGEs during 14 days of exposure. Moreover, fructose caused more protein carbonyl (PCO) formation and also oxidized thiol groups more in glycated than in native BSA. The extract of C. rotundus at different concentrations (25-250microg/ml) has significantly decreased the formation of AGEs in term of the fluorescence intensity of glycated BSA. Furthermore, we demonstrated the significant effect of C. rotundus extract on preventing oxidative protein damages including effect on PCO formation and thiol oxidation which are believed to form under the glycoxidation process. Our results highlight the protein glycation inhibitory and antioxidant activity of C. rotundus. These results might lead to the possibility of using the plant extract or its purified active components for targeting diabetic complications.


Angiogenesis and tumor grading in primary breast cancer patients: an analysis of 158 needle core biopsies.
Angiogenesis, the formation of new blood vessels from a preexisting vascular bed, is a complex multistep process. To investigate how tumor angiogenesis correlates with tumor grading in breast carcinoma diagnosed on core biopsy, microvessels were counted (and graded the density of microvessels) within the initial invasive carcinomas of 158 patients. Using light microscopy, the number of microvessels was counted manually in a subjectively selected hot spot (in the most active areas of neovascularization per 400x field), and their values were separated as above or below median (low and high), without knowledge of the outcome in the patient or any other pertinent variable. When the mean values of MVD of the various groups defined by tumor grading were compared, significant difference was noted (P=2.61E-17). When the mean values of MVD of the groups defined by tumor grading risk were compared, significant difference was noted (P=7.68E-05). When tumors were classified as high or low MVD, based on a cut-off value (30.70175 microvessels/mm2), cases with high MVD were significantly more numerous in MSBR four patients. MVD did show a relationship with groups defined by MSBR grade (P=3.25209E-07) or with tumor grading risk (P=2.54181E-06). Assessment of tumor angiogenesis may therefore prove valuable in selecting patients with early breast carcinoma for aggressive therapy.


"Smart" liposomal nanocontainers in biology and medicine.
The perspectives of using liposomes for delivery of drugs to desired parts of the human body have been intensively investigated for more than 30 years. During this time many inventions have been suggested and different kinds of liposomal devices developed, and a number of them have reached the stages of preclinical or clinical trials. The latest techniques can be used to develop biocompatible nano-sized liposomal containers having some abilities of artificial intellect, such as the presence of sensory and responsive units. However, only a few have been clinically approved. Further improvements in this area depend on our knowledge of the interactions of drugs with the lipid bilayer of liposomes. Further studies on liposomal transport through the human body, their targeting of cells requiring therapeutic treatment, and finally, the development of techniques for controlled drug delivery to desired acceptors on cell surfaces or in cytoplasm are still required.


Activation of the Arp2/3 complex by the Listeria acta protein. Acta binds two actin monomers and three subunits of the Arp2/3 complex.
ActA is a bacterially encoded protein that enables Listeria monocytogenes to hijack the host cell actin cytoskeleton. It promotes Arp2/3-dependent actin nucleation, but its interactions with cellular components of the nucleation machinery are not well understood. Here we show that two domains of ActA (residues 85-104 and 121-138) with sequence similarity to WASP homology 2 domains bind two actin monomers with submicromolar affinity. ActA binds Arp2/3 with a K(d) of 0.6 microm and competes for binding with the WASP family proteins N-WASP and Scar1. By chemical cross-linking, ActA, N-WASP, and Scar1 contact the same three subunits of the Arp2/3 complex, p40, Arp2, and Arp3. Interestingly, profilin competes with ActA for binding of Arp2/3, but actophorin (cofilin) does not. The minimal Arp2/3-binding site of ActA (residues 144-170) is C-terminal to both actin-binding sites and shares sequence homology with Arp2/3-binding regions of WASP family proteins. The maximal activity at saturating concentrations of ActA is identical to the most active domains of the WASP family proteins. We propose that ActA and endogenous WASP family proteins promote Arp2/3-dependent nucleation by similar mechanisms and require simultaneous binding of Arp2 and Arp3.


Comparative genome analyses of Serratia marcescens FS14 reveals its high antagonistic potential.
S. marcescens FS14 was isolated from an Atractylodes macrocephala Koidz plant that was infected by Fusarium oxysporum and showed symptoms of root rot. With the completion of the genome sequence of FS14, the first comprehensive comparative-genomic analysis of the Serratia genus was performed. Pan-genome and COG analyses showed that the majority of the conserved core genes are involved in basic cellular functions, while genomic factors such as prophages contribute considerably to genome diversity. Additionally, a Type I restriction-modification system, a Type III secretion system and tellurium resistance genes are found in only some Serratia species. Comparative analysis further identified that S. marcescens FS14 possesses multiple mechanisms for antagonism against other microorganisms, including the production of prodigiosin, bacteriocins, and multi-antibiotic resistant determinants as well as chitinases. The presence of two evolutionarily distinct Type VI secretion systems (T6SSs) in FS14 may provide further competitive advantages for FS14 against other microbes. To our knowledge, this is the first report of comparative analysis on T6SSs in the genus, which identifies four types of T6SSs in Serratia spp.. Competition bioassays of FS14 against the vital plant pathogenic bacterium Ralstonia solanacearum and fungi Fusarium oxysporum and Sclerotinia sclerotiorum were performed to support our genomic analyses, in which FS14 demonstrated high antagonistic activities against both bacterial and fungal phytopathogens.


Effects of metabolism inhibitors, antiestrogens and antiandrogens on the androgen and estrogen induced sexual behavior in Japanese quail.
The relative contribution of androgenic and estrogenic metabolites of testosterone to the activation of sexual behavior was studied in Japanese quail by using inhibitors of testosterone metabolism, antiestrogens and antiandrogens. These compounds were tested in castrated birds whose sexual behavior had been activated by silastic implants of testosterone (T) or daily injections of testosterone propionate (TP) or diethylstilboestrol (DES). The aromatase inhibitor ATD only depressed T-induced behavior when injected at high doses and the 5 alpha-reductase inhibitor, 4MA was inactive in this respect. The antiestrogens, tamoxifen (TAM) and nitromifene citrate (CI-628) strongly depressed sexual behavior but they also drastically reduced the crowing behavior which is typically androgen-dependent which throws some doubts on the specificity of their action. The antiandrogens, flutamide and cyproterone acetate (CA), only had limited inhibitory effects on the copulatory behavior but similarly decreased only marginally the crowing. As they strongly depressed the cloacal gland growth, it can be ascertained that they were injected in sufficient amounts and their lack of action on crowing questions the ability of these compounds to inhibit brain processes even when they are androgen-dependent. Taken together with the results of previous experiments which tested the behavioral effects of the testosterone metabolites, the present data confirm the implication of both androgenic and estrogenic metabolites of testosterone in the activation of behavior. Their interaction remains, however, poorly defined and its understanding will probably require the identification of the biochemical processes which in the brain mediate the behavior.


A Novel Treatment for Glomerular Disease: Targeting the Activated Macrophage Folate Receptor with a Trojan Horse Therapy in Rats.
Since activated macrophages express a functional folate receptor β (FRβ), targeting this macrophage population with folate-linked drugs could increase selectivity to treat inflammatory diseases. Using a macrophage-mediated anti-glomerular basement membrane (anti-GBM) glomerulonephritis (GN) in WKY rats, we investigated the effect of a novel folic acid-aminopterin (AMT) conjugate (EC2319) designed to intracellularly deliver AMT via the FR. We found that treatment with EC2319 significantly attenuated kidney injury and preserved renal function. Kidney protection with EC2319 was blocked by a folate competitor, indicating that its mechanism of action was specifically FRβ-mediated. Notably, treatment with methotrexate (MTX), another folic acid antagonist related to AMT, did not protect from kidney damage. EC2319 reduced glomerular and interstitial macrophage infiltration and decreased M1 macrophage recruitment but not M2 macrophages. The expression of CCL2 and the pro-fibrotic cytokine TGF-β were also reduced in nephritic glomeruli with EC2319 treatment. In EC2319-treated rats, there was a significant decrease in the deposition of collagens. In nephritic kidneys, FRβ was expressed on periglomerular macrophages and macrophages present in the crescents, but its expression was not observed in normal kidneys. These data indicate that selectively targeting the activated macrophage population could represent a novel means for treating anti-GBM GN and other acute crescentic glomerulonephritis.


Studies of the mortality of atomic bomb survivors, Report 14, 1950-2003: an overview of cancer and noncancer diseases.
This is the 14th report in a series of periodic general reports on mortality in the Life Span Study (LSS) cohort of atomic bomb survivors followed by the Radiation Effects Research Foundation to investigate the late health effects of the radiation from the atomic bombs. During the period 1950-2003, 58% of the 86,611 LSS cohort members with DS02 dose estimates have died. The 6 years of additional follow-up since the previous report provide substantially more information at longer periods after radiation exposure (17% more cancer deaths), especially among those under age 10 at exposure (58% more deaths). Poisson regression methods were used to investigate the magnitude of the radiation-associated risks, the shape of the dose response, and effect modification by gender, age at exposure, and attained age. The risk of all causes of death was positively associated with radiation dose. Importantly, for solid cancers the additive radiation risk (i.e., excess cancer cases per 10(4) person-years per Gy) continues to increase throughout life with a linear dose-response relationship. The sex-averaged excess relative risk per Gy was 0.42 [95% confidence interval (CI): 0.32, 0.53] for all solid cancer at age 70 years after exposure at age 30 based on a linear model. The risk increased by about 29% per decade decrease in age at exposure (95% CI: 17%, 41%). The estimated lowest dose range with a significant ERR for all solid cancer was 0 to 0.20 Gy, and a formal dose-threshold analysis indicated no threshold; i.e., zero dose was the best estimate of the threshold. The risk of cancer mortality increased significantly for most major sites, including stomach, lung, liver, colon, breast, gallbladder, esophagus, bladder and ovary, whereas rectum, pancreas, uterus, prostate and kidney parenchyma did not have significantly increased risks. An increased risk of non-neoplastic diseases including the circulatory, respiratory and digestive systems was observed, but whether these are causal relationships requires further investigation. There was no evidence of a radiation effect for infectious or external causes of death.


A well-oxygenated cells environment may help to fight against protein glycation.
Normoglycemic Wistar rats' Glycated Hemoglobin Levels (GHL) showed a time-dependent difference between control groups and those exposed to regular inhalation of peroxidizing extracts of turpentine. These extracts were able to optimize the oxygen permeation at the cellular level during and subsequently to a breathing session. The more the rats breathed turpentine peroxidized vapor, the lower their GHL was. This study was designed to confirm, in ex-vivo blood samples, the impact of peroxidizing extract on the GHL. Red blood cells were separated from plasmas by centrifugation. Plasmas were treated by peroxidizing and non-peroxidizing turpentine vapor or untreated (control), then combined with washed red blood cells three hours before evaluation. Glycation of hemoglobin proteins was quantified according to the Habeed's method. The ex-vivo experiments showed that the peroxidizing terpenes reduced the GHL after a three-hour contact. So did oxidized terpenes. Controls and the volatile component of the expended essential oil showed the opposite results. Optimal oxygenation, especially when facilitated by the peroxidized volatile component of the essential oil of turpentine, can protect organisms (mammals in this study) from protein glycation. Optimizing oxygenation can also reduce the GHL of treated blood samples after three hours of incubation.


Sex pili mutants isolated by macarbomycin treatment.
Macarbomycin (MM) preferentially kills Escherichia coli carrying episomes derepressed for sex pili formation (drd episomes). Using MM, we have isolated various types of mutants from both F-lac.tet and R100-1 episomes and have classified them into two types by sex pili formation. Among 26 mutants, 20 could not produce pili and had lost both the ability for conjugal transmission and sensitivity to male phages. From 12 of these 20 mutants, revertants capable of pili formation could be obtained. Most of these had simultaneously regained susceptibility to MM, indicating that the MM susceptibility of E. coli carrying drd episomes is due to pili formation. Three of the revertants could not confer MM susceptibility on their host and two of them could not produce normal pili, indicating that there are some types of pili insensitive to MM. The six MM-resistant mutants could produce pili, but the frequency of pili formation was slightly lower than that of their parent episomes. Mutants of this type exhibited various degrees of decrease in conjugal transferability or male phage sensitivity, and one mutant produced pili that were altered in shape. It is suggested that this latter mutant became MM-resistant as a result of changes in the chemical composition of its pili.


[Nursing auxiliaries and technicians and hospital infection control in surgical centers: myths and truths].
This analytical and descriptive study is aimed at detecting concepts that translate myths and truths on hospital infection among nursing auxiliaries and technicians at the surgical centers of three hospitals. The data collection instrument consisted of 28 affirmative statements (15 true and 13 false) encompassing factors related to the patient, the surgical team, the environment, and procedures. The statements contain a three-point scale (I agree, I am in doubt, I disagree). We received 72% of adequate answers and 28% of non-adequate, which indicates that perioperative nursing professionals have satisfactory knowledge of hospital infection control. In the items use of safety footwear, rings and other objects, hair as a pathogen, hand scrubbing, use of humid gown and surgical area, infected surgery and cleaning routine, occupational diseases, hospital infection, surgical site infection and surgery time we were able to detect myths and rituals about infection control, which are mainly related to the culture of those who practice them, thus perpetuating resistance to change.


Expression of growth-related genes and drug-resistance genes in HTLV-I-positive and HTLV-I-negative post-thymic T-cell malignancies.
This study was designed to investigate the biologic and molecular basis of the aggressive behavior of high-grade post-thymic T-cell malignancies. Freshly frozen tumor tissues from (1) human T-cell leukemia/lymphoma virus type I (HTLV-I)-positive adult T-cell lymphoma (ATL) (7 cases), (2) HTLV-I-negative aggressive T-cell lymphoma (12 cases), and (3) HTLV-I-negative nonaggressive T-cell lymphoma (11 cases) were studied for the expression of several growth-related genes or proliferation antigens including interleukin-2 receptor (IL-2R), Ki-67, transforming growth factor-beta (TGF-beta), topoisomerase, and the multidrug resistance (MDR) gene by immunohistochemistry and Northern blot hybridization. Our results showed that tumor cells associated with HTLV-I and anaplastic morphology had an enhanced expression of Ki-67, TGF-beta, and topoisomerase, as compared to nonaggressive T-cell lymphoma. The expression of IL-2R was limited to ATL and one Ki-1 lymphoma. The MDR gene was frequently expressed in ATL, but only infrequently in other, HTLV-I-negative, malignancies. Clinical progression or relapse was associated with the expression of MDR, in addition to an increased expression of Ki-67. We therefore conclude that the aggressive clinical behavior of high-grade T-cell lymphoma may result mainly from the high proliferative activity of tumor cells, but the association with HTLV-I and clinical relapse is further complicated by the development of drug resistance.


[Diagnostic value of autoantibodies to asialoglycoprotein receptor in autoimmune hepatitis].
Among 106 patients with autoimmune liver disease in 12 (11.3%) were found autoantibodies to asialoglycoprotein receptor (anti-ASGPR): in 5 (9.1%) of 55 with PBC, and 4 (17.4%) of 23 with AIH and 3 (10.7%) of 28 patients with syndrome chiasm (PBC + AIH). Detection of antibodies against the ASGPR is an effective method for diagnosis of autoimmune hepatitis. Further improvement of this method allows for monitoring and disease. Anti-ASGPR levels correlate with biochemical parameters and with the severity and manifestation autoimmune processes in patients with autoimmune hepatitis.


Complete Closed Genome Sequences of Three Bibersteinia trehalosi Nasopharyngeal Isolates from Cattle with Shipping Fever.
Bibersteinia trehalosi is a respiratory pathogen affecting cattle and related ruminants worldwide. B. trehalosi is closely related to Mannheimia haemolytica and is often associated with bovine respiratory disease complex (BRDC), a polymicrobial multifactorial disease. We present three complete closed genome sequences of this species generated using an automated assembly pipeline.


A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death.
Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine. We present a family in which a common polymorphism (single nucleotide polymorphism) inherited from the father, combined with a stop codon mutation inherited from the mother (both asymptomatic), led to 2 cases of sudden infant death. KCNQ1, KCNH2, SCN5A, KCNE1, KCNE2, CACNA1c, CACNB2b, and KCNJ2 genes were amplified and analyzed by direct sequencing. Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary (CHO-K1) and COS-1 cells. An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis. The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc (560 ms). The mother was asymptomatic but displayed a prolonged QTc. Genetic screening of the mother revealed a heterozygous nonsense mutation (P926AfsX14) in KCNH2, predicting a stop codon. The father was asymptomatic with a normal QTc but had a heterozygous polymorphism (K897T) in KCNH2. The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14. Heterologous coexpression of K897T and P926AfsX14 led to loss of function of HERG current much greater than expression of K897T or P926AfsX14 alone. Our data suggest that a common polymorphism (K897T) can markedly accentuate the loss of function of mildly defective HERG channels, leading to long-QT syndrome-mediated arrhythmias and sudden infant death.


Infundibular Pulmonic Stenosis in a Moluccan Cockatoo (Cacatua moluccensis).
A 31-year-old female Moluccan cockatoo (Cacatua moluccensis) was examined for intermittent foot clenching of 4 months' duration. Physical examination revealed feather-destructive behavior and clinical findings compatible with hypovitaminosis A. Neurologic examination was unremarkable. Results of radiographs, hematologic testing, plasma biochemical analyses, and measurement of lead and trace element blood concentrations were unremarkable, except for degenerative joint disease of several thoracic intervertebral joints and a low blood copper concentration. Increased dietary copper was recommended. After a 6-month period without clinical signs, the bird presented again for episodes of foot weakness. Radiographic review was suggestive of mild pulmonary trunk enlargement. Echocardiography revealed mild mitral and aortic regurgitation, dilation of the ascending aorta, and a dilated right ventricle with turbulent right ventricular outflow. An electrocardiogram revealed a sinus rhythm and normal-appearing complexes. Nonselective fluoroscopic angiography was performed 3 weeks later because of persistent episodes of foot clenching and weakness. Infundibular pulmonic stenosis, poststenotic dilation of the pulmonic trunk, and proximal main pulmonary arteries were identified, as well as a mild narrowing of the descending aorta compatible with aortic stenosis. The bird was discharged without medication but with dietary recommendations and experienced 2 clenching episodes in the days after the last visit. No recurrence of clinical signs has been reported over the 18-month follow-up period. To our knowledge, this is the first report of infundibular pulmonic stenosis in a bird. This case illustrates the application of basic and advanced diagnostic imaging modalities in evaluating cardiac disease in birds.


Cyanophage-encoded lipid desaturases: oceanic distribution, diversity and function.
Cyanobacteria are among the most abundant photosynthetic organisms in the oceans; viruses infecting cyanobacteria (cyanophages) can alter cyanobacterial populations, and therefore affect the local food web and global biochemical cycles. These phages carry auxiliary metabolic genes (AMGs), which rewire various metabolic pathways in the infected host cell, resulting in increased phage fitness. Coping with stress resulting from photodamage appears to be a central necessity of cyanophages, yet the overall mechanism is poorly understood. Here we report a novel, widespread cyanophage AMG, encoding a fatty acid desaturase (FAD), found in two genotypes with distinct geographical distribution. FADs are capable of modulating the fluidity of the host's membrane, a fundamental stress response in living cells. We show that both viral FAD (vFAD) families are Δ9 lipid desaturases, catalyzing the desaturation at carbon 9 in C16 fatty acid chains. In addition, we present a comprehensive fatty acid profiling for marine cyanobacteria, which suggests a unique desaturation pathway of medium- to long-chain fatty acids no longer than C16, in accordance with the vFAD activity. Our findings suggest that cyanophages are capable of fiddling with the infected host's membranes, possibly leading to increased photoprotection and potentially enhancing viral-encoded photosynthetic proteins, resulting in a new viral metabolic network.


5-Aza-2'-Deoxycytidine Alters the Methylation Profile of Bortezomib-Resistant U266 Multiple Myeloma Cells and Affects Their Proliferative Potential.
Multiple myeloma (MM) is a plasma cell malignancy that accounts for 1% of all cancers and is the second-most-common hematological neoplasm. Bortezomib (BTZ) is a proteasome inhibitor widely implemented in the treatment of MM alone or in combination with other agents. The development of resistance to chemotherapy is one of the greatest challenges of modern oncology. Therefore, it is crucial to discover and implement new adjuvant therapies that can bypass therapeutic resistance. In this paper, we investigated the in vitro effect of methylation inhibitor 5-Aza-2'-deoxycytidine on the proliferative potential of MM cells and the development of resistance to BTZ. We demonstrate that alterations in the DNA methylation profile are associated with BTZ resistance. Moreover, the addition of methylation inhibitor 5-Aza-2'-deoxycytidine to BTZ-resistant MM cells led to a reduction in the proliferation of the BTZ-resistant phenotype, resulting in the restoration of sensitivity to BTZ. However, further in vitro and ex vivo studies are required before adjuvant therapy can be incorporated into existing treatment regimens.


Peak heart rates at extreme altitudes.
We have measured maximal heart rate during a graded maximal bicycle exercise test to exhaustion in five healthy climbers before and during an expedition to Mt. Everest. Maximal heart rates at sea level were 186 (177-204) beats/min(-1) at sea level and 170 (169-182) beats/min(-1) with acute hypoxia. After 1, 4 and 6 weeks of acclimatization to 5400 m, maximal heart rates were 155 (135-182), 158 (144-182), and 155 (140-183) beats/min(-1), respectively. Heart rates of two of the climbers were measured during their attempt to reach the summit of Mt. Everest without the use of supplemental oxygen. The peak heart rates at 8,750 m for the two climbers were 142 and 144 beats/min(-1), which were similar to their maximal heart rates during exhaustive bicycle exercise at 5,400 m, the values being 144 and 148 beats/min(-1), respectively. The peak heart rates at 8,750 m are in agreement with other field studies, but considerably higher than values reported from hypobaric chamber studies.


In vivo metabolism of leukotriene C4 in germ-free and conventional rats. Fecal excretion of N-acetylleukotriene E4.
[5,6,8,9,11,12,14,15-3H8]Leukotriene C4 was subcutaneously injected into rats. Substantial amounts of the administered radioactivity were excreted in feces of germ-free and conventional animals during a 72-h period (78 and 64%, respectively). Analyses of fecal extracts by high performance liquid chromatography showed eight radioactive components for each type of animal. One metabolite amounted to 4.6% of the injected radioactivity in germ-free and 0.6% in conventional rats. Its chemical structure, 5-hydoxy-6-S-(2-acetamido-3-thiopropionyl)-7,9-trans-11,14-c is-eicosatetraenoi c acid (N-acetylleukotriene E4) was determined by ultraviolet spectroscopy, fast atom bombardment mass spectrometry, chemical and enzymatic transformations, and confirmed by chemical synthesis. Another metabolite (2.7% of the administered radioactivity in germ-free and 0.5% in conventional rats) was characterized as the 11-trans isomer of the former metabolite. The pathway of formation of these compounds appears to be analogous to the pathway of mercapturic acid biosynthesis.


Hydrogel-Based Macroscopic Click Chemistry.
The click reaction has found good utility across various fields due to the characteristics of high efficiency, atom economy, simple and mild reaction conditions. Click chemistry is usually utilized for connecting components of microscopic level, while it is still unable for joining macroscopic building blocks. Materials consisting of macroscopic building blocks realize the flexible fabrication of three-dimensional structures at macroscopic level, exerting significance on parallel manufactures. In this work, we reported macroscopic click chemistry utilizing hydrogel as macroscopic building blocks. Hydrogels G1 and G2 were prepared by incorporating M1 (N,N'-dimethyl-1,2-ethanediamine) and P1 (alkyne functionalized polyethylene glycol) respectively, where polymer chains formed through diffusion-induced amino-yne click reaction entangled different hydrogel networks together. Additionally, chain-like aggregates and complicated 3D structures such as tetrahedron and quadrangular pyramid were constructed based on the adhesion of the hydrogel blocks. The approach enables us to find more possibilities in the delicate designation of 3D aggregations as well as large-scale manufacturing.


Complications following volar locking plate fixation for distal radial fractures: a systematic review.
The purpose of this systematic review is to assess the prevalence of complications following volar locking plate fixation of distal radial fractures. A computer-based search was carried out using EMBASE and PUBMED/MEDLINE. Only prospective comparative and prospective cohort studies that presented data concerning complications after treatment of distal radial fractures with a volar locking plate in human adults with a minimal follow-up of 6 months were included. Two quality assessment tools were used to assess the methodological quality of the studies (level of evidence rating according to the Oxford Centre of Evidence Based Medicine and the modified version of the Cochrane Bone, Joint and Muscle Trauma Group's former quality assessment tool). Thirty three studies were eligible for final assessment. Most complications were problems with nerve and tendon function as well as complex regional pain syndrome. With an overall complication rate of 16.5%, most of which were 'minor' complications and low rates of nonunion and malunion, volar locking plate fixation can be considered a reasonably safe treatment option for patients with distal radial fractures.


Anxiety Disorders: A Review.
Anxiety disorders have a lifetime prevalence of approximately 34% in the US, are often chronic, and significantly impair quality of life and functioning. Anxiety disorders are characterized by symptoms that include worry, social and performance fears, unexpected and/or triggered panic attacks, anticipatory anxiety, and avoidance behaviors. Generalized anxiety disorder (6.2% lifetime prevalence), social anxiety disorder (13% lifetime prevalence), and panic disorder (5.2% lifetime prevalence) with or without agoraphobia are common anxiety disorders seen in primary care. Anxiety disorders are associated with physical symptoms, such as palpitations, shortness of breath, and dizziness. Brief screening measures applied in primary care, such as the Generalized Anxiety Disorder-7, can aid in diagnosis of anxiety disorders (sensitivity, 57.6% to 93.9%; specificity, 61% to 97%). Providing information about symptoms, diagnosis, and evidence-based treatments is a first step in helping patients with anxiety. First-line treatments include pharmacotherapy and psychotherapy. Selective serotonin reuptake inhibitors (SSRIs, eg, sertraline) and serotonin-norepinephrine reuptake inhibitors (SNRIs, eg, venlafaxine extended release) remain first-line pharmacotherapy for generalized anxiety disorder, social anxiety disorder, and panic disorder. Meta-analyses suggest that SSRIs and SNRIs are associated with small to medium effect sizes compared with placebo (eg, generalized anxiety disorder: standardized mean difference [SMD], -0.55 [95% CI, -0.64 to -0.46]; social anxiety disorder: SMD, -0.67 [95% CI, -0.76 to -0.58]; panic disorder: SMD, -0.30 [95% CI, -0.37 to -0.23]). Cognitive behavioral therapy is the psychotherapy with the most evidence of efficacy for anxiety disorders compared with psychological or pill placebo (eg, generalized anxiety disorder: Hedges g = 1.01 [large effect size] [95% CI, 0.44 to 1.57]; social anxiety disorder: Hedges g = 0.41 [small to medium effect] [95% CI, 0.25 to 0.57]; panic disorder: Hedges g = 0.39 [small to medium effect[ [95% CI, 0.12 to 0.65]), including in primary care. When selecting treatment, clinicians should consider patient preference, current and prior treatments, medical and psychiatric comorbid illnesses, age, sex, and reproductive planning, as well as cost and access to care. Anxiety disorders affect approximately 34% of adults during their lifetime in the US and are associated with significant distress and impairment. First-line treatments for anxiety disorders include cognitive behavioral therapy, SSRIs such as sertraline, and SNRIs such as venlafaxine extended release.


A case of Korean patient with macular corneal dystrophy associated with novel mutation in the CHST6 gene.
To report a novel mutation within the CHST6 gene, as well as describe light and electron microscopic features of a case of macular corneal dystrophy. A 59-year old woman with macular corneal dystrophy in both eyes who had decreased visual acuity underwent penetrating keratoplasty. Further studies including light and electron microscopy, as well as DNA analysis were performed. Light microscopy of the cornea revealed glycosaminoglycan deposits in the keratocytes and endothelial cells, as well as extracellularly within the stroma. All samples stained positively with alcian blue, colloidal iron, and periodic acid-Schiff. Electron microscopy showed keratocytes distended by membrane-bound intracytoplasmic vacuoles containing electron-dense fibrillogranular material. These vacuoles were present in the endothelial cells and between stromal lamellae. Some of the vacuoles contained dense osmophilic whorls. A novel homozygous mutation (c.613 C>T [p.Arg205Trp]) was identified within the whole coding region of CHST6. A novel CHST6 mutation was detected in a Korean macular corneal dystrophy patient.


Nitrogen source activates TOR (target of rapamycin) complex 1 via glutamine and independently of Gtr/Rag proteins.
The evolutionary conserved TOR complex 1 (TORC1) activates cell growth in response to nutrients. In yeast, TORC1 responds to the nitrogen source via a poorly understood mechanism. Leucine, and perhaps other amino acids, activates TORC1 via the small GTPases Gtr1 and Gtr2, orthologs of the mammalian Rag GTPases. Here we investigate the activation of TORC1 by the nitrogen source and how this might be related to TORC1 activation by Gtr/Rag. The quality of the nitrogen source, as defined by its ability to promote growth and glutamine accumulation, directly correlates with its ability to activate TORC1 as measured by Sch9 phosphorylation. Preferred nitrogen sources stimulate rapid, sustained Sch9 phosphorylation and glutamine accumulation. Inhibition of glutamine synthesis reduces TORC1 activity and growth. Poor nitrogen sources stimulate rapid but transient Sch9 phosphorylation. A Gtr1 deficiency prevents the transient stimulation of TORC1 but does not affect the sustained TORC1 activity in response to good nitrogen sources. These findings suggest that the nitrogen source must be converted to glutamine, the preferred nitrogen source in yeast, to sustain TORC1 activity. Furthermore, sustained TORC1 activity is independent of Gtr/Rag. Thus, the nitrogen source and Gtr/Rag activate TORC1 via different mechanisms.


Down-regulation of miR-125a-5p is associated with salivary adenoid cystic carcinoma progression via targeting p38/JNK/ERK signal pathway.
Salivary adenoid cystic carcinoma (SACC) is a relatively uncommon epithelial-like malignancy that can occur in the head and neck region. Despite its slow growth, this aggressive salivary gland tumor frequently recurs and metastasizes to distant organs since lacking effective chemotherapy treatment. MicroRNAs are key regulators in tumor metastasis and progression, but their roles during SACC progression have not been illustrated. In current study, we demonstrate that miR-125a-5p is down-regulated in SACC and closely related to the metastasis and progression in human SACC specimens. In vitro, miR-125a-5p mimic can suppress SACC cell migration and invasion; while blocking miR-125a-5p can relieve the inhibition effect. By using dual-luciferase assay, we confirmed that miR-125a-5p directly targeted to p38 and tissue samples of patients indicated the negative correlation between miR-125a-5p and p38; clinical analysis also showed that low level expression of miR-125a-5p is closely associated with poor prognosis of SACC. Furthermore, down-regulation of miR-125a-5p triggered downstream p38/JNK/ERK activation. Taken together, our results indicate that down-regulation of miR-125a-5p promotes SACC progression through p38 signal pathway and miR-125a-5p can be a potential therapeutic target of SACC.


The flexor carpi radialis H-reflex in polyneuropathy: relations to conduction velocities of the median nerve and the soleus H-reflex latency.
In 80 controls latencies of flexor carpi radialis (FCR) and in 94 controls latencies of soleus H-reflexes correlated well with length of the extremity, body height and age. Multiple regression equations using latency as a variable dependent on age and body height can be best used in practice when both reflexes are employed for demonstration of proximal pathology. The majority (69%) of 93 patients with various polyneuropathies showed abnormalities in both reflexes illustrating that proximal nerve segments are frequently involved. Four per cent had abnormal FCR H-reflexes with normal soleus H-reflexes whereas the reverse was found in 19% of the patients. Abnormal FCR H-reflexes occurred with normal motor and sensory conduction velocities in the peripheral part of the median nerve in 14%, whereas the reverse was seen in 12%, indicating that FCR H-reflex examination is a valuable supplement to conventional conduction studies for detection of electrophysiologically existing pathology.


Aztreonam plus piperacillin--empiric treatment of neutropenic fever in gynecology-oncology patients receiving cisplatin-based chemotherapy.
Antibiotic therapy must be instituted promptly and on an empiric basis in neutropenic patients. We evaluated the efficacy of a combined antibiotic regimen of monobactam (aztreonam) and antipseudomonal penicillin (piperacillin) in treating neutropenic fever episodes in gynecologic-oncology patients receiving cisplatin-based chemotherapy. A retrospective analysis of response to this regimen was performed. The rationale of this combination is the lack of nephrotoxicity and ototoxicity in patients who are or were previously treated with other nephrotoxic/ototoxic agents like cisplatin. A total of 19 courses of this regimen was administered to 13 patients with neutropenic fever following a complete fever work-up. Aztreonam (1-2gr q8h) plus piperacillin (4gr q8h) were administered intravenously for 6-8 days. Blood cultures were positive in four febrile episodes, and urine cultures were positive in seven. Gram negative organisms accounted for all positive cultures. The cultured organism showed in-vitro sensitivity to at least one of the drugs in all positive isolates. Clinical response with defervescence was noted during therapy in 18/19 courses (94.7%). Although the two drugs share a common bactericidal mechanism they were found to be highly active in this subgroup of patients. A double blind prospective evaluation of this empiric combination is warranted.


Reusable Self-Sterilization Masks Based on Electrothermal Graphene Filters.
Surgical mask is recommended by the World Health Organization for personal protection against disease transmission. However, most of the surgical masks on the market are disposable that cannot be self-sterilized for reuse. Thus, when confronting the global public health crisis, a severe shortage of mask resource is inevitable. In this paper, a novel low-cost electrothermal mask with excellent self-sterilization performance and portability is reported to overcome this shortage. First, a flexible, ventilated, and conductive cloth tape is patterned and adhered to the surface of a filter layer made of melt-blown nonwoven fabrics (MNF), which functions as interdigital electrodes. Then, a graphene layer with premier electric and thermal conductivity is coated onto the MNF. Operating under a low voltage of 3 V, the graphene-modified MNF (mod-MNF) can quickly generate large amounts of heat to achieve a high temperature above 80 °C, which can kill the majority of known viruses attached to the filter layer and the mask surface. Finally, the optimized graphene-modified masks based on the mod-MNF filter retain a relatively high particulate matter (PM) removal efficiency and a low-pressure drop. Moreover, the electrothermal masks can maintain almost the same PM removal efficiency over 10 times of electrifying, suggesting its outstanding reusability.


Bioavailability of zinc in milk and soy protein-based infant formulas.
Total femur zinc of young rats was used to evaluate the biological availability of zinc in milk and soy protein-based infant formulas. A zinc deficient diet (0.8 mug Zn/g) containing egg white protein was supplemented with graded levels of zinc from zinc sulfate, milk and soy protein-based infant formulas. A plot of total femur zinc (log) after feeding the diet for 3 weeks versus the zinc added to the diet gave a linear relationship over the range of 0, 3, 6, 9 and 12 mug/g added zinc. By using a slope-ratio bioassay model, the relative biological availability of endogenous and added zinc in milk-based formula was estimated to be 0.86 and that of soy-based formula 0.67 (zinc sulphate = 1.00) with corresponding 95% fiducial limits being 0.82 to 0.91 and 0.62 to 0.71. Thus, to provide equivalent amounts of available zinc, the total zinc content of the soy protein-based formula would need to be at least 20% higher than that of the formula containing milk protein.


Day-90 survival in critically-ill patients with COVID-19 and hydroxychloroquine: a propensity analysis.
There are limited data on the effect of hydroxychloroquine on medium term outcomes in patients with coronavirus disease 2019 (COVID-19) requiring intensive care. We aimed to evaluate the effects of hydroxychloroquine on day 90 mortality in this specific population. This retrospective, multicenter, propensity matched cohort analysis, used data of adult patients with laboratory confirmed COVID-19 admitted to 3 university affiliated intensive care units between March 7, 2020, to April 7, 2020 in Lyon, France. Patients received either hydroxychloroquine (loading dose of 400 mg twice daily at day 1 followed by 200 mg twice daily from day 2 to day 10) or standard of care without hydroxychloroquine. We compared all-cause mortality at day-90 after ICU admission between propensity score matched groups receiving hydroxychloroquine or standard of care. A total of 157 patients were included with a day-28 and day-90 mortality rate of 23.6% and 32.5%, respectively. The median (interquartile) age was 67 years (56-76 years), 105 (66.9%) were men, 65 (41.4%) fulfilled criteria for acute respiratory distress syndrome, and 64 (41%) received hydroxychloroquine (HCQ) for 10 days (4-10 days). In the propensity score matched cohort (59 patients in each group), day-90 mortality was 35.6% for patients who received HCQ and 23.7% for patients who did not (P=0.23). Kaplan Meier survival analysis showed no statistically significant association between HCQ therapy and mortality (P=0.20 by log-rank test). In this study, off-label use of HCQ in critically ill patients with COVID-19 was not associated with any significant change in medium-term prognosis, confirming results of studies in less severe patients.


Total RNA Sequencing of Rett Syndrome Autopsy Samples Identifies the M4 Muscarinic Receptor as a Novel Therapeutic Target.
Mutations in the MeCP2 gene are responsible for the neurodevelopmental disorder Rett syndrome (RTT). MeCP2 is a DNA-binding protein whose abundance and ability to complex with histone deacetylase 3 is linked to the regulation of chromatin structure. Consequently, loss-of-function mutations in MeCP2 are predicted to have broad effects on gene expression. However, to date, studies in mouse models of RTT have identified a limited number of gene or pathway-level disruptions, and even fewer genes have been identified that could be considered amenable to classic drug discovery approaches. Here, we performed RNA sequencing (RNA-seq) on nine motor cortex and six cerebellar autopsy samples from RTT patients and controls. This approach identified 1887 significantly affected genes in the motor cortex and 2110 genes in the cerebellum, with a global trend toward increased expression. Pathway-level analysis identified enrichment in genes associated with mitogen-activated protein kinase signaling, long-term potentiation, and axon guidance. A survey of our RNA-seq results also identified a significant decrease in expression of the CHRM4 gene, which encodes a receptor [muscarinic acetylcholine receptor 4 (M4)] that is the subject of multiple large drug discovery efforts for schizophrenia and Alzheimer's disease. We confirmed that CHRM4 expression was decreased in RTT patients, and, excitingly, we demonstrated that M4 potentiation normalizes social and cognitive phenotypes in Mecp2+/- mice. This work provides an experimental paradigm in which translationally relevant targets can be identified using transcriptomics in RTT autopsy samples, back-modeled in Mecp2+/- mice, and assessed for preclinical efficacy using existing pharmacological tool compounds.


[A case study of shifting a patient with percutaneous endoscopic gastrostomy (PEG) to home care].
We studied the primary factor preventing a patient with PEG from being transferred to home care focusing on the dietic situation. The study has revealed differences in not the age of the patient, the days of hospitalization or hematological and biochemical test data but in the condition of use of home care services or family makeup. In other words, insufficient use of social resources is considered to prevent shifting to home care or continuation of home care of patients with PEG. It is necessary to understand not only dietic situation but social backgrounds of patients and supply information.