Queuine metabolism and cadmium toxicity in Drosophila melanogaster.
Queuine can replace guanine in the anticodon of certain tRNAs and is a hypermodified guanine derivative that can be synthesized by bacteria but not by mice. The study demonstrates that Drosophila can incorporate dietary queuine into tRNA but cannot synthesize it de novo for this purpose. Since an earlier study had shown that dietary CdCl2 caused Drosophila to increase greatly the proportion of queuine-containing tRNA over non-queuine tRNA the ability of dietary queuine to counteract cadmium toxicity was evaluated. When queuine was present in the cadmium-containing medium more pupae matured into adults than when queuine was absent. Other studies had demonstrated that the transglycosylase enzyme, that catalyzes the replacement of guanine in the anticodon of tRNA by queuine, is present in Drosophila larvae but the tRNA is virtually devoid of queuine. This study shows that in the presence of dietary queuine the larval tRNA contains abundant amounts of queuine. Therefore, we postulate a significant role for bacteria in supplying queuine to Drosophila for its incorporation into tRNA and that the control of this process by Drosophila is passive, i.e. is not an essential feature in differentiation.


Site-specific cleavage by metal ion cofactors and inhibitors of M1 RNA, the catalytic subunit of RNase P from Escherichia coli.
The location of phosphate residues involved in specific centers for binding of metal ions in M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli, was determined by analysis of induction of cleavage of RNA by metal ions. At pH 9.5, Mg2+ catalyzes cleavage of M1 RNA at five principal sites. Under certain conditions, Mn2+ and Ca2+ can each replace Mg2+ as the cofactor in the processing of precursor tRNAs by M1 RNA and P RNA, the RNA subunit of RNase P from Bacillus subtilis. These cations, as well as various metal ion inhibitors of the catalytic activity of M1 RNA, also promote cleavage of M1 RNA in a specific manner. Certain conditions that affect the catalytic activity of M1 RNA also alter the rate of metal ion-induced cleavage at the various sites. From these results and a comparison of cleavage of M1 RNA with that of a deletion mutant of M1 RNA and of P RNA, we have identified two different centers for binding of metal ions in M1 RNA that are important for the processing of the precursor to tRNA(Tyr) from E. coli. There is also a center for the binding of metal ions in the substrate, close to the site of cleavage by M1 RNA.


Incidence of non-A, non-B hepatitis after screening blood donors for antibodies to hepatitis C virus and surrogate markers.
To compare the effect of screening blood donors for antibodies to hepatitis C virus (anti-HCV) on the incidence of non-A, non-B hepatitis in recipients with that of screening blood donors for antibodies to hepatitis B core antigen (anti-HBc) and elevated alanine aminotransferase levels. Cohort analysis of serum samples from donors and recipients. Recipients were followed for 12 months to determine the occurrence of non-A, non-B hepatitis. The blood-transmitted viruses unit and the liver unit of a university teaching hospital. A total of 250 patients who had open heart surgery and their 3142 blood donors. Donor sera were tested for anti-HCV by enzyme-linked immunosorbent assay (ELISA) and, in the event of a positive result, by recombinant immunoblot assay (RIBA). Antibodies to anti-HBc and serum alanine aminotransferase (ALT) levels were also measured. Measurements of anti-HCV and ALT activity in recipients were done before transfusion and at regular intervals during follow-up. Of the 250 transfusion recipients, 40 developed non-A, non-B hepatitis. Of the 3142 donors, 70 were positive for anti-HCV by ELISA, 440 were positive for anti-HBc, and 177 had alanine aminotransferase levels between 0.67 and 1.33 mukat/L. The sensitivity (87%), specificity (89%), positive predictive value (59%), and negative predictive value (97%) of blood-donor screening were higher for anti-HCV than for anti-HBc (82%, 36%, 21%, and 91%, respectively) and 70%, 29%, and 91%, respectively). The expected number of donors excluded because of the presence of anti-HCV was considerably smaller than that of donors with positive results for surrogate markers of hepatitis. Screening blood donors for the presence of anti-HCV is more accurate than screening for surrogate markers (anti-HBc and ALT) and protects more effectively against post-transfusion non-A, non-B hepatitis.


Periodontal vaccine: A dream or reality.
Recent advances in cellular and molecular biology have led to the development of new strategies for vaccines against many types of infectious diseases. It has long been recognized that individuals who recovered from a disease developed subsequent resistance to the same. In the late 18th century, Edward Jenner developed and established the principle of vaccination using the cross protection conferred by cowpox virus, which is non-pathogenic in humans. With the rapid growth of microbial genome sequencing and bioinformatics analysis tools we have the potential to examine all the genes and proteins from any human pathogen. This technique has the capability to provide us with new targets for anti-microbial drugs and vaccines. However, to realize this potential new bioinformatics and experimental approaches to select these targets from the myriad of available candidates are required. Vaccination is a process that induces specific immune resistance to a bacterial or viral infection.


Protein kinase C activation precedes arachidonic acid release during IgG-mediated phagocytosis.
Previous studies demonstrated that both protein kinase C (PKC) and arachidonic acid (AA) are required for IgG-mediated phagocytosis by human monocytes. We have characterized a calcium-independent "phagocytic" phospholipase A2 (designated pPL) that mediates arachidonic acid release. The present studies were designed to order PKC and pPL in the phagocytic signaling pathway. The PKC inhibitors staurosporine and calphostin C caused a coordinated decrease in phagocytosis of IgG-opsonized erythrocytes and arachidonic acid release. The PLA2 activators mastoparan and melittin restored phagocytosis to PKC-inhibited cells, but were ineffective in monocytes pretreated with the pPL inhibitor bromoenol lactone. Similarly, PKC activation with PMA and diacylglycerol enhanced phagocytosis in the absence, but not in the presence, of bromoenol lactone. These results indicate that pPL may be regulated by an upstream phosphorylation event. Thus, we examined the effects of Ab-opsonized glass bead ingestion, okadaic acid-mediated inhibition of phosphatases, and PMA treatment on the activity of pPL and on its distribution between the cytosolic and membrane-associated compartments. IgG-opsonized erythrocytes and okadaic acid caused an overall increase in pPL activity, with a twofold increase in membrane-associated pPL. PMA treatment caused a 1.8-fold increase in membrane-associated pPL activity. Okadaic acid and PMA mimic IgG-opsonized erythrocytes with respect to membrane activation of pPL, suggesting that pPL activity may be regulated by PKC. Collectively, these results indicate that pPL activity is modulated by PKC during IgG-mediated phagocytosis, and that the PKC requirement can be bypassed by direct activation of pPL.


Worker notification: lessons from the past.
The methodological choices being made in current worker notification programs are influenced in both obvious and subtle ways by certain ground-breaking notifications carried out during the 1970s and 1980s. A lore has built up concerning these cases. Their names are heavy with implied meaning. As worker notification gains in acceptance and frequency, some of those newly interested in the area will be unfamiliar with these early cases, their significance, and their lessons. To help provide a context for these proceedings, six landmark worker notifications are briefly described here. Different cases might have been chosen; more cases might have been added. However, these six give a flavor of the struggles, trials, triumphs, and insights that have marked worker notification's passage through recent history.


Clinicopathological and Radiological Features with Long Term Follow Up of Metaplastic Carcinoma Breast in India.
Metaplastic breast carcinoma (MBC) is a heterogeneous group of invasive carcinomas with squamous and/or mesenchymal differentiation. Because of their rare occurrence, the information regarding the clinical behaviour of metaplastic carcinomas is limited. The purpose of our study was to delineate the clinicopathological and radiological features, treatment outcomes, prognostic factors, and survival of patients with MBC. Ambispective observational study with prospective recruitment was done from 1st January 2019 to 31st August 2020. Retrospective data included between 1st January 2009 and 31st December 2018. In the retrospective group surgical database of our department was searched and those with MBC diagnosis on post-operative histopathology recruited. In prospective group patients with MBC on core biopsy were followed and those operated were included. The patients followed up at our breast cancer clinic (BCC) and their demographic, clinical, pathological radiological and treatment details noted. Forty patients formed the study population. The mean age of the patents was 42 years. Ipsilateral axillary lymph node metastasis was present in 22.5%. The pathological median tumor size was 5.4 (range 2.1 to 22 cm). The most common differentiation was cartilaginous (35%) followed by squamous (32.5%). The most common mammographic grading was BIRADS 4 (Breast Imaging Reporting and Data system). Magnetic resonance imaging was T2 hyperintense with peripheral rim enhancement and restriction on DWI. The median overall (OS) and disease-free survival (DFS) was 42 and 40 months, respectively. Fifteen patients (37.5%) had disease related mortality. A subgroup analysis revealed that, type of differentiation, histopathology and tumor size > 5cm affected both OS and DFS significantly. Metaplastic breast cancer in our setup presents in young patients with aggressive large tumors at a higher stage and diverse histopathology and with comparable overall and disease-free survival. The histological subtype, tumor differentiation and tumor size are prognostic factors.


[Determination of 11S IgA in nasal secretions in the monitoring of local immunomodulating therapy].
Twenty pediatric patients with recurrent infectious diseases of the upper respiratory tract (tonsillitis, adenotonsillitis with or without involvement of the ear and/or lower respiratory tract) were studied. An immunological assay of the nasal secretum was performed at time of diagnosis and following treatment with a local immunomodulator drug, administered by spray. The 7S, 11S IgA and albumin rates were evaluated. The authors emphasize the importance of SIgA in mucose defense mechanisms as well as the need for a selective method for determining the 11S IgA level. An original method for immuno-isoelectrofocusing (IIEF) determination was employed in the present study. After treatment a significant increase in 11S IgA level was observed, especially in those patients with a significant basal SIGA deficit. The authors describe details of the technique for determination and discuss the results.


[Are age and educational level the determinants of hormonal parameters considered as indices of andropause?].
The involutionary processes in gonadal and adrenal glands are significant for male aging. The dynamics of hormonal changes in aging men seems to be individually differentiated and vary in distinct populations. Currently there are no data on social differences in hormonal parameters in men. The study was carried out in order to evaluate the age-related changes of hormonal parameters considered as indices of andropause and analyze the social gradients in these variables in healthy Polish men. Material comprised the data of 414 healthy men, inhabitants of Wrocław, aged 32-79, examined in 2000 in DOLMED (Wrocław, Lower Silesia). Serum levels of the following hormones were assessed using radioimmunological assays: free testosterone (FT), total testosterone (TT), estradiol (E2), dehydroepiandrosterone sulphate (DHEAS), luteinizing hormone (LH), sex hormone binding globulin (SHBG) and insulin-like growth factor 1 (IGF-1). The following indices were calculated: FAI (Free Androgen Index), TT/E2 and TT/LH. Among Polish men there were negative correlations between age and serum levels of FT, DHEAS, IGF-1, E2, and between age and FAI, TT/LH. The correlation between the DHEAS level and age was the strongest among all relationships here. The male aging was accompanied by the serum reduction, during 5 years: FT level of 4.8%, DHEAS--8.6% and IGF-1--5.0%. The reduction in serum DHEAS and IGF-1 levels reached 77.3% and 44.9% (respectively) between men aged 32-34 and the oldest subjects aged 75-79. The decrease in serum FT level between men aged 32-34 and those aged 70-74 was 38.2%. The relationships between age and E2 level and between age and TT/LH were not strong (respectively, beta = -0.10, p < 0.05 and beta = -0. 17, p < 0.001). The male aging was also accompanied by the increase in serum SHBG (11.7%/5 years) and LH levels (13.5%/5 years). Among male there were no age-related changes in the serum TT level and TT/E2. The results of a two-way ANOVA revealed that education significantly differentiated serum IGF-1 levels (independently on age). The highest IGF-1 levels were observed in men who had graduated from university, the lowest--in those who had finished the trade school at the very most. In contrast, there were no social differences in other analysed hormonal parameters.


[Three-Step Interdisciplinary Team Program for Nasogastric Tube Care].
Patients with chewing disorders and dysphagia often take nutrition via nasogastric tube feeding. Patients with indwelling nasogastric tubes often experience discomfort, resulting in increased incidences of self-extubation and slippage. Frequent changes or misplacement of the nasogastric tube may also cause patient safety issues such as aspiration pneumonia, gastrointestinal trauma, and pneumothorax. Collaboration within the interdisciplinary team is needed to provide proper nasogastric tube care prior to placement, during the indwelling process, and during the removal of the nasogastric tube. Interdisciplinary teams should develop standardized procedures for nasogastric tube care, initiate swallow screening, oral care and training early, and develop a care plan for nasogastric tube removal. An evidence-based, practical example of nasogastric tube care implemented by an interdisciplinary team in a neurology unit is presented in this article. Interdisciplinary teams may assist patients to prepare to return to a normal tube-free life using individualized, safe, and dignified nasogastric tube care to improve quality of life.


Mixed-Haplotype and Mixed-Isotype Major Histocompatibility Complex Class II Molecules Detected by Murine T-Cell Clones and Their Possible Involvement in Autoimmunity
The recognition of antigen by T lymphocytes (T cells) is restricted by Class I or Class II major histocompatibility complex (MHC) gene products, the phenomenon called "MHC restriction." MHC restriction is speculated to be one of the major elements for the association of disease susceptibility to MHC haplotypes. Clones of T cells have been shown to be powerful tools for the analysis of such restriction specificity. In this report, I describe unique mixed-isotype Abetad/Ealphad restriction molecules detected by T-cell clones in (B6Ealphad x BALB/c)F1 transgenic mice. The restriction specificity of these clones was confirmed by anti-Class II mAb blocking experiments. The ability of spleen cells from Abetad and Ealphad double transgenic B6 (B6AbetadEalphad) mice that express Abetad/Ealphad molecules to present KLH to these clones supported the existence of such unique specificity. I also describe autoreactive as well as KLH-reactive T-cell clones restricted by mixed-haplotype Abetaz/Aalphad Class II molecules derived from (NZB x NZW)F1 (B/WF1) mice. The restriction specificity was demonstrated by mAb blocking experiments and by experiments using Class II gene-transfected antigen-presenting cells. It is possible that such unique mixed-isotype and mixed-haplotype Class II molecules are critically involved in autoimmunity. In addition, the detailed methodology for establishing T-cell clones currently employed in my laboratory is described.


Double-outlet right ventricle with extremely rare conotruncal morphology.
We describe an extremely rare form of double-outlet right ventricle in a 1-year-old boy who had l-malposition of the great arteries with the definitive conotruncal features of false Taussig-Bing heart. Extraordinary conotruncal morphology raised several surgical options, however, the definitive anatomical repair was achieved by an arterial switch operation with a baffle from a ventricular septal defect to the pulmonary artery.


Lymph node yield as a prognostic factor in clinically node negative oral cavity squamous cell carcinoma.
To determine the prognostic significance of nodal yield in patients with clinically node-negative (cN0) oral cavity squamous cell carcinoma (OCSCC). This retrospective observational study included 40 patients with cN0 OCSCC who underwent treatment with at least 6 months of follow-up data from November 2012 to April 2020. We recorded the variables, including patient demographics, cancer site, tumor-node-metastasis (TNM) staging, type of treatment, lymph node yield (LNY), histopathologic diagnosis, and recurrence. The recorded data were analyzed with descriptive and interferential statistics using specific tests. Our study cohort comprised of 27 males and 13 females with a mean age of 60.08+13.153 years. Tongue (55%) was the commonly affected site. Seventeen (42.5%) patients belonged to TNM stage II. The mean LNY in our study was 38.65±25.41 (range 7-98). Following surgery, 19 (47.5%) patients further received adjuvant therapies. Recurrence was reported only in 4 (10%) patients. There was no significant difference between LNY and recurrence rate (p=0.892). Factors including, age (p=0.121), gender (p=0.209), site (p=0.519), size of tumor (p=0.416) did not influence the LNY. There is no correlation between LNY and recurrence in cN0 OCSCC patients in our study. Meticulous neck dissection and thorough pathologic reporting prevents TNM under staging and improves the overall survival and prognosis.


Optimization of laccase production by Trametes versicolor cultivated on industrial waste.
Laccases are very interesting biocatalysts for several industrial applications. Its production by different white-rot fungi can be stimulated by a variety of inducing substrates, and the use of lignocellulosic wastes or industrial by-products is one of the possible approaches to reduce production costs. In this work, various industrial wastes were tested for laccase production by Trametes versicolor MZKI G-99. Solid waste from chemomechanical treatment facility of a paper manufacturing plant showed the highest potential for laccase production. Enzyme production during submerged cultivation of T. versicolor on the chosen industrial waste has been further improved by medium optimization using genetic algorithm. Concentrations of five components in the medium were optimized within 60 shake-flasks experiments, where the highest laccase activity of 2,378 U dm(-3) was achieved. Waste from the paper industry containing microparticles of CaCO(3) was found to stimulate the formation of freely dispersed mycelium and laccase production during submerged cultivation of T. versicolor. It was proven to be a safe and inexpensive substrate for commercial production of laccase and might be more widely applicable for metabolite production by filamentous fungi.


Future treatment of peptic ulcers.
Drugs which abolish acid secretion and promise improved ulcer healing rates have produced dysplasia and tumours in experimental animals given large doses for long periods. Solutions to this problem will be based on better understanding of the mechanisms involved. Healing rates might be improved by combining inhibitors of acid secretion with ulcer protectors which, unlike currently available agents, are designed to work well in the presence of diminished acid secretion. Drugs which increase mucosal defence might best be used to prevent ulceration, so the need is to develop agents which are safe for long term use.


Effects of feeding beef females supplemental fat during gestation on cold tolerance in newborn calves.
Effects of prepartum fat supplementation of the dam on cold tolerance of calves were determined in two studies. In Exp. 1, 22 F1, crossbred heifers gestating F2 calves received diets containing either 1.7 or 4.7% dietary fat starting at d 230+/-2d of gestation. Safflower seeds (Carthamus tinctorius) containing 37% oil with 79% linoleic acid were the supplemental fat source in isocaloric-isonitrogenous diets. Calves were separated from their dams at birth, fed pooled dairy-cow colostrum, muzzled to prevent sucking, and returned to their dams in a heated (22 degrees C) barn for 3.5 h. At 4 h of age, a jugular catheter was inserted. At 5 h of age, calves were placed in a 0 degrees C room for 140 min and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for glucose, cortisol, and cholesterol. In Exp. 2, 18 multiparous, crossbred beef cows bred to Murray Grey sires were randomly assigned to receive diets containing either 1.7 or 3.1% dietary fat starting at 235+/-2 d gestation. Safflower seeds were used as the supplemental fat source in isocaloric-isonitrogenous diets. At d 260 of gestation, premature parturition was induced in one-half of the cows from each diet group by feeding Ponderosa pine (Pinus ponderosa) needles. Experimental protocols were the same as in Exp. 1, except that cold exposure was at 9 degrees C for 200 min. Rectal temperatures were affected in Exp. 1 by time and diet x time (both P < .01) and diet x calf sex (P < .05) and in Exp. 2 by calf age (P < .05), time, and calf age x time (both P < .01). Plasma cortisol concentrations were affected by time (P < .01) and calf sex x time (P < .05) in Exp. 1 and by time ( P < .01) in Exp. 2. Cholesterol concentrations in Exp. 1 were affected by diet x time (P < .05) and in Exp. 2 by time (P < .05). Plasma glucose concentrations were affected in Exp. 1 by diet (P < .05) and in Exp. 2 by calf age, time, and calf age x time (all P < .01). We conclude from Exp. 1 that feeding heifers supplemental fat during late gestation increased glucose concentrations in the newborn calf, resulting in favorable responses in body temperature in the cold-stressed newborns. This increase in substrate availability suggests a potential positive effect on heat generation in newborns during sustained periods of cold stress. In Exp. 2, premature calves had compromised cold tolerance possibly due to impaired shivering or brown adipose tissue thermogenesis.


Chemical composition and in-vitro antimicrobial activity of the essential oils of three Greek Achillea species.
The chemical composition of the essential oils of Achillea holosericea, Achillea taygetea, Achillea fraasii was determined by GC/MS analysis. Among the ninety-five assayed constituents, camphor, borneol and 1,8-cineol were found to be the major components. The in-vitro antimicrobial activity of these essential oils was evaluated against six bacteria indicating that the first is totally inactive, while the other two possess moderate to strong activities mainly against the Gram negative strains. The essential oil of A. fraasii was also active against the tested pathogenic fungi.


Dental status as a quality control health care parameter for children with disabilities.
The aim of this study was to determine the level of dental health condition in children with disabilities and to find out weather dental health status might be used as a quality control parameter regarding overall health care for disabled children. Disabled and healthy children from 3 to 17 years old were examined. There were 86 boys and 34 girls in each group. Dental health status was evaluated using the World Health Organization diagnostic criteria for decayed, missing and filled teeth. The relations between mean decayed, missing and filled teeth index for primary, mixed and permanent dentition showed no statistically significant differences among groups. Our results showed that disabled children have evenly level of dental caries as their healthy peers. One can conclude that organized health care for disabled children have positive influence on dental care, too. Furthermore, dental status as an indicator of dental care level might be used as a parameter for quality control regarding overall health care for disabled children.


Parkinson's disease and dementia with neuronal inclusions in the cerebral cortex: Lewy bodies or Pick bodies.
Cortical Lewy bodies may be difficult to differentiate from Pick bodies by the usual staining methods. This problem is illustrated in a case of progressive dementia with parkinsonian features. Lewy bodies were found, not only in the pigmented nuclei in the brainstem and in the cerebral cortex, but also in the dentate fascia, a predilection site for Pick bodies. The inclusion bodies were compared with the inclusion bodies in a case of Pick's disease. Intense argyrophilia of the cortical inclusion bodies argued in favor of Pick bodies; antibodies to phosphorylated neurofilaments reacted with the cortical inclusions and with classical Pick bodies, but antibodies to paired helical filaments reacted only with the Pick bodies. The most convincing evidence, that the inclusions were Lewy bodies, was obtained by electron microscopy. The filaments in the inclusions showed fuzzy deposits of electron dense material, characteristic for filaments of Lewy bodies. This contrasted with the smooth filaments of the Pick bodies. It is concluded that cortical inclusions in brains from patients with Lewy bodies in the pigmented nuclei of the brainstem most likely represent Lewy bodies. This can be confirmed by examining the ultrastructure of the inclusions.


Quantitative production of compound I from a cytochrome P450 enzyme at low temperatures. Kinetics, activation parameters, and kinetic isotope effects for oxidation of benzyl alcohol.
Cytochrome P450 enzymes are commonly thought to oxidize substrates via an iron(IV)-oxo porphyrin radical cation transient termed Compound I, but kinetic studies of P450 Compounds I are essentially nonexistent. We report production of Compound I from cytochrome P450 119 (CYP119) in high conversion from the corresponding Compound II species at low temperatures in buffer mixtures containing 50% glycerol by photolysis with 365 nm light from a pulsed lamp. Compound I was studied as a reagent in oxidations of benzyl alcohol and its benzylic mono- and dideuterio isotopomers. Pseudo-first-order rate constants obtained at -50 degrees C with concentrations of substrates between 1.0 and 6.0 mM displayed saturation kinetics that gave binding constants for the substrate in the Compound I species (K(bind)) and first-order rate constants for the oxidation reactions (k(ox)). Representative results are K(bind) = 214 M(-1) and k(ox) = 0.48 s(-1) for oxidation of benzyl alcohol. For the dideuterated substrate C(6)H(5)CD(2)OH, kinetics were studied between -50 and -25 degrees C, and a van't Hoff plot for complexation and an Arrhenius plot for the oxidation reaction were constructed. The H/D kinetic isotope effects (KIEs) at -50 degrees C were resolved into a large primary KIE (P = 11.9) and a small, inverse secondary KIE (S = 0.96). Comparison of values extrapolated to 22 degrees C of both the rate constant for oxidation of C(6)H(5)CD(2)OH and the KIE for the nondeuterated and dideuterated substrates to values obtained previously in laser flash photolysis experiments suggested that tunneling could be a significant component of the total rate constant at -50 degrees C.


Fluorescent substrates useful as high-throughput screening tools for ADAM9.
Fluorescence resonance energy transfer substrates were designed and tested as substrates for ADAM9. The donor/quencher pair used were 5-carboxy fluorescein (Fam) and 4-(4-dimethyl-aminophenylazo)benzoyl (Dabcyl) since they have been well studied sensitive fluorescent probes. The peptides based on precursor TNF-alpha, Dabcyl-Ser-Pro-Leu-Ala-Gln-Ala-Val-Arg-Ser-Ser-Lys(Fam)- NH2 and Dabcyl-Leu-Ala-Gln-Ala-HomoPhe-Arg-Ser-Lys(Fam)- NH2, and C-terminal TGF-alpha, Dabcyl-Glu-His-Ala-Asp-Leu-Leu-Ala-Val-Val-Ala-Ala-Lys(Fam)- NH2 cleavage sites were effectively processed by ADAM9 with turnover numbers of 100 +/- 20 x 10(-2) min(-1), 20 +/- 10 x 10(-2) min(-1), and 10 +/- 3 x 10(-2) min(-1). In addition, a peptide based on the 33 kDa cleavage site of the low affinity receptor for IgE, CD23, Dabcyl-Leu-Arg-Ala-Glu-Gln-Gln-Arg-Leu-Lys-Ser-Lys(Fam)- NH2 was processed as well but with less efficiency. A more selective substrate for ADAM9 was found based on the betacellulin cleavage site. However, the valine containing precursor TNF-alpha based substrate was used to measure IC50 values of metalloproteinase inhibitors against ADAM9 since it was processed the most efficiently. The tightest binding inhibitor was the Wyeth Aerst compound, TMI-1, with an IC50 of 2.1 +/- 0.3 nM. In addition, GI254023, previously identified as a selective inhibitor of ADAM10, also inhibited ADAM9 with an IC50 of 280 +/- 110 nM. These results demonstrate that sensitive substrates for ADAM9 can be developed that are useful in high-throughput screening assays for ADAM9.


Temperature dependent hysteresis-like changes of motility parameters in human spermatozoa.
We studied the impact of temperature changes on selected parameters of normozoospermic spermatozoa motility. The examinations were carried out in the temperature range of 11 degrees C to 21 degrees C both during cooling and heating. We found that a phenomenon of hysteresis, i.e. alternate means of changes in velocity straight linear and lateral head displacement was obtained both at cooling and at heating. This phenomenon was not found for other parameters examined.


Graphical contig analyzer for all sequencing platforms (G4ALL): a new stand-alone tool for finishing and draft generation of bacterial genomes.
Genome assembly has always been complicated due to the inherent difficulties of sequencing technologies, as well the computational methods used to process sequences. Although many of the problems for the generation of contigs from reads are well known, especially those involving short reads, the orientation and ordination of contigs in the finishing stages is still very challenging and time consuming, as it requires the manual curation of the contigs to guarantee correct identification them and prevent misassembly. Due to the large numbers of sequences that are produced, especially from the reads produced by next generation sequencers, this process demands considerable manual effort, and there are few software options available to facilitate the process. To address this problem, we have developed the Graphic Contig Analyzer for All Sequencing Platforms (G4ALL): a stand-alone multi-user tool that facilitates the editing of the contigs produced in the assembly process. Besides providing information on the gene products contained in each contig, obtained through a search of the available biological databases, G4ALL produces a scaffold of the genome, based on the overlap of the contigs after curation. THE SOFTWARE IS AVAILABLE AT: http://www.genoma.ufpa.br/rramos/softwares/g4all.xhtml.


Facts, fallacies, and politics of comparative effectiveness research: Part 2 - implications for interventional pain management.
The United States leads the world in many measures of health care innovation. However, it has been criticized to lag behind many developed nations in important health outcomes including mortality rates and higher health care costs. The surveys have shown the United States to outspend all other Organisation for Economic Co-operation and Development (OECD) countries with spending on health goods and services per person of $7,290 - almost 2(1/2) times the average of all OECD countries in 2007. Rising health care costs in the United States have been estimated to increase to 19.1% of gross domestic product (GDP) or $4.4 trillion by 2018. CER is defined as the generation and synthesis of evidence that compares the benefits and harms of alternate methods to prevent, diagnose, treat, and monitor a clinical condition or to improve the delivery of care. The, comparative effectiveness research (CER) has been touted by supporters with high expectations to resolve most ill effects of health care in the United States providing high quality, less expensive, universal health care. The efforts of CER in the United States date back to the late 1970s and it was officially inaugurated with the enactment of the Medicare Modernization Act (MMA). It has been rejuvenated with the American Recovery and Reinvestment Act (ARRA) of 2009 with an allocation of $1.1 billion. CER has been the basis of decision for health care in many other countries. Of all the available agencies, the National Institute for Health and Clinical Excellence (NICE) of the United Kingdom is the most advanced, stable, and has provided significant evidence, though based on rigid and proscriptive economic and clinical formulas. While CER is taking a rapid surge in the United States, supporters and opponents are emerging expressing their views. Since interventional pain management is a new and evolving specialty, with ownership claimed by numerous organizations, at times it is felt as if it has many fathers and other times it becomes an orphan. Part 2 of this comprehensive review will provide facts, fallacies, and politics of CER along with discussion of potential outcomes, impact of CER on health care delivery, and implications for interventional pain management in the United States.


Effects of LH and PGF2alpha in equine dominant follicles observed by MDS.
Microdialysis System (MDS) is a novel technique used for investigation of molecule secretion between different cell populations. Local hormonal secretion at follicular wall has been still unclear. This MDS study was used to determine progesterone (P4), androstenedione (A4), estradiol-17beta (E2) and Prostaglandin F2alpha (PGF2alpha) release in mare pre-ovulatory follicles. Follicles larger than 30 mm were isolated from the ovary and follicular fluid aspirated for hormone assay. Follicular fluid collected from small, middle and large follicles were analyzed by EIA. The concentrations of P4 and PGF2alpha were similar among the different sizes of follicles. The release of A4 was observed in middle and large follicles. E2 concentration was observed in middle follicles and was higher in large follicles compared with middle follicles. Follicular wall was cut and incubated for MDS and when LH was infused, there was an increase in P4 and A4 release. PGF2alpha release was considerably high after LH infusion compared to the control group. Infusion of PGF2alpha increased P4 and A4 release but there was no change in E2 release. This results suggest that in pre-ovulatory follicles, LH stimulates theca interna cells and also PGF2alpha seemed to have a mediator role to induce steroid hormone production and luteinization of follicular cells. The nature of the mechanisms involved in selection of large follicles is still a perplexing research problem in reproduction.


Sperm morphology training in India.
Sperm morphology is regarded as possibly the most consistent sperm variable that is related to in vitro and in vivo fertilisation success or failure. There is also a re-emerging awareness of andrology, particularly in developing countries where healthcare resources are often limited. This study aimed to record the importance of hands-on training on the sperm morphology reading skills among 174 Indian infertility workers. During a series of five workshops presented by the Indian Academy of Clinical Embryologists, delegates were requested to record the percentage normal spermatozoa on pre-stained Diff Quick slides. A total of four slides each containing different percentages normal spermatozoa were used during the workshops. The delegates were requested to record the percentage normal spermatozoa prior and after training. The results were expressed as the percentage difference between the reference value and the pre-training and post-training values reported by the workshop delegates. The percentage difference indicated a significant improvement in the reading skills, for example from 253 ± 244% to 20 ± 93% for slide 1. Similar values were recorded for slide 2, 3 and 4. If we consider the present findings as representative of medical laboratory scientist's sperm morphology reading skills, clinicians should be concerned about the diagnostic quality as far as male infertility is concerned.


Dendrimer drugs prevent scar tissue formation.
Extract: Dendrimers are hyperbranched synthetic macromolecules that can be made with structural precision. So far, their biomedical applications have been limited to drug delivery. Dendrimers with cationic (i.e., amine or NH3+) end groups are toxic after repeated intravenous use or topical ocular application. In contrast, anionic (i.e., carboxy or COO-) dendrimers are not toxic. For example, cationic dendrimers cause substantial changes to red blood cell morphology at 10 mug/ml whilst anionic dendrimers have no such effect at 2,000 mug/ml. We set out to determine whether the receptor-ligand interactions between carbohydrates and proteins, which mediate many important aspects of cell surface-mediated immuno-regulation could be pharmacologically manipulated using new anionic dendrimer-based drugs. Any such medicine would have to possess multiple and cooperative receptor binding properties. Previous attempts to pharmacologically manipulate such interactions with synthetic linear polymers have failed because of the structural diversity of the polymers used, and because they activated immunological cell death pathways and blood clotting pathways.


Photoaffinity labeling of a cell surface polyamine binding protein.
Intracellular polyamine pools are partially maintained by an active transport apparatus that is specific for and regulated by polyamines. Although mammalian transport activity has been characterized by kinetic studies, the actual protein itself has yet to be identified, purified, or cloned. As one approach to this problem, we attempted photoaffinity labeling of plasma membrane proteins using two specifically designed and synthesized polyamine conjugates as photoprobes. The first is a spermidine conjugate bearing the photoreactive moiety 4-azidosalicylic acid at the N4 position via an alkyl linkage, and the second is a norspermine conjugate with 4-azidosalicylic acid at the N4 position via an acyl linkage. Labeling of murine L1210 lymphocytic leukemia cells was carried out at 4 degrees C to promote selective alkylation of cell surface proteins. Separation of plasma membrane proteins from cells cross-linked with the N4-spermidine conjugate by SDS-polyacrylamide gel electrophoresis revealed two heavily labeled proteins at approximately 118 and approximately 50 kDa (designated p118 and p50, respectively). Band p118 was more well defined and much more intensely labeled. Analogous proteins were also observed in human U937 lymphoma cells. Specificity of labeling was strongly suggested by competition with polyamines and analogs during labeling and further indicated by the nearly identical labeling of the same protein by the N1-norspermine photoprobe but not by the unconjugated photoreagent. Neuraminidase pretreatment of L1210 cells increased mobility of the p118, suggesting that it was glycosylated and, thus, of plasma membrane origin. In transport-deficient L1210 cells, p118 and p50 were found to have a slightly higher molecular mass and were accompanied by a less distinct protein band (approximately 100 kDa). These findings indicate the presence of a polyamine binding protein at the surface of murine and human leukemia cells, which could be directly or indirectly related to the polyamine transport apparatus.


A NMR and theoretical study of the aggregates between alkyllithium and chiral lithium amides: control of the topology through a single asymmetric center.
The complexes between methyllithium and chiral 3-aminopyrrolidine (3-AP) lithium amides bearing a second asymmetric center on their lateral amino group were studied using multinuclear ((1)H, (6)Li, (13)C, (15)N) low-temperature NMR spectroscopies in tetrahydrofuran-d(8). The results indicate that lithium chelation forces the pyrrolidine ring of the 3-AP to adopt a norbornyl-like conformation and that robust 1:1 noncovalent complexes between methyllithium and 3-AP lithium amides form in the medium. A set of (1)H-(1)H and (1)H-(6)Li NMR cross-coupling correlations shows that the binding of methyllithium can take place along the "exo" or the "endo" face of this puckered structure, depending on the relative configuration of the lateral chiral group. This aggregation step renders the nitrogen of the 3-amino group chiral, the "exo" and "endo" topologies corresponding to the (S) and (R) configurations, respectively, of this atom. Density functional theory calculations show that the "exo" and "endo" arrangements are, for both diastereomers, almost isoenergetic even when solvent is taken into account. This result suggests that the formation of the mixed aggregates is under strict kinetic control. A relationship between the topology of these complexes and the sense of induction in the enantioselective alkylation of aromatic aldehydes by alkyllithiums is proposed.


Effect of Maitland mobilization and exercises for the treatment of shoulder adhesive capsulitis: a single-case design.
The purpose of this single-case design (ABCBC) was to investigate the response of shoulder motions, pain, and function to two commonly used physiotherapy management approaches. An individual with stage three shoulder adhesive capsulitis was treated with exercise (phase B) and exercise plus mobilization (phase C). Initially, a "baseline" phase (phase A) when treatment had not started was established for comparison. Two types of Maitland "accessory" glenohumeral mobilization techniques, anteroposterior mobilization in shoulder flexion and longitudinal caudad in shoulder abduction, were evaluated during phase C. The Shoulder Pain and Disability Index (SPADI) was used to monitor pain and functional disability, and four shoulder movements (flexion, abduction, internal, and external rotations) were measured. The results were evaluated by using single-case design analysis method of Split Middle Technique and visual observation. The SPADI scores deteriorated in phase A but improved in phase B1, C1, and B2. All four shoulder movements improved under both management approaches, although more gain in motion was observed when mobilizations were added to an exercise program. The exercise plus mobilization intervention shows promise as a cost-effective management. The deterioration in shoulder motion, pain, and function observed in phase A may suggest benefit of an earlier physiotherapy intervention.


Assessing smoke-free workplaces in Wisconsin municipal and county government buildings, 1997.
Passive smoke has been shown to cause cancer and heart disease in humans. We surveyed all Wisconsin cities and villages in 1997 to determine their clean indoor air policies in government-owned buildings. By 1997, 62% of cities and villages and 100% of counties had enacted a clean indoor air ordinance or policy. Smoking is completely banned in 26% of all city and village buildings and 49% of county buildings in the state. This survey shows that an increasing proportion of government worksites in Wisconsin are enacting clean indoor air policies to protect their employees and the public from the health effects of passive smoke.


Investigations on the morphophysiological basis of revascularization of the myocardium by the Vineberg method.
A study was carried out on the dynamics of vasculogenesis and the formation of mammary-coronary anastomoses within the first seven months after implantation of the internal mammary artery in the left ventricular myocardium, according to the Vineberg method. The efficiency of these anastomoses was demonstrated 4 to 7 months later by the absence of lesions 8 to 10 days after transitory and permanent myocardial ischaemia induced by coronary obliteration. The lesions could not be prevented in the case of thrombosis of the internal mammary artery or when ischaemia was produced concomitantly with the implant or before the mammary-coronary arteriolo-arterial anastomotic network developed.


No evidence for local adaptation and an epigenetic underpinning in native and non-native ruderal plant species in Germany.
Many invasive species have rapidly adapted to different environments in their new ranges. This is surprising, as colonization is usually associated with reduced genetic variation. Heritable phenotypic variation with an epigenetic basis may explain this paradox.Here, we assessed the contribution of DNA methylation to local adaptation in native and naturalized non-native ruderal plant species in Germany. We reciprocally transplanted offspring from natural populations of seven native and five non-native plant species between the Konstanz region in the south and the Potsdam region in the north of Germany. Before the transplant, half of the seeds were treated with the demethylation agent zebularine. We recorded survival, flowering probability, and biomass production as fitness estimates.Contrary to our expectations, we found little evidence for local adaptation, both among the native and among the non-native plant species. Zebularine treatment had mostly negative effects on overall plant performance, regardless of whether plants were local or not, and regardless of whether they were native or non-native. Synthesis. We conclude that local adaptation, at least at the scale of our study, plays no major role in the success of non-native and native ruderal plants. Consequently, we found no evidence yet for an epigenetic basis of local adaptation.


Fluorescence analysis of biochemical constituents identifies atherosclerotic plaque with a thin fibrous cap.
Vulnerable plaque generally contains a thin fibrous cap, lipid pools, and reduced internal plaque collagen. Arterial fluorescence analysis can differentiate atherosclerotic lesions from normal arteries; however, the contribution of the lipid core to atherosclerotic arterial fluorescence remains controversial. This study aimed to identify lipid core fluorophores and to differentiate the lipid core from normal artery and atheroma. The helium-cadmium laser-induced fluorescence spectra of cadaveric arteries and known chemical constituents were recorded. Lipid core fluorescence spectra exhibited marked red shifts and broadening compared with the fluorescence spectra of normal tissue and atheroma. Similar fluorescence spectra were obtained for lipid core and oxidized low density lipoprotein, for atheroma and collagen, and for normal artery and elastin. A classification based on collagen, elastin, and oxidized low density lipoprotein spectral decomposition could discriminate the lipid core (n=29), normal artery (n=74), atheroma (n=73), and preatheroma (n=10) with 86% accuracy. Fibrous cap thickness was correlated with the spectral collagen content index (r=0.65, P<0.0001), especially at a thickness of <200 microm. We conclude that a classification algorithm based on chemical spectral decomposition can accurately classify the fluorescence spectra of normal artery, atheroma, and lipid core and may be useful in identifying vulnerable atheroma in vivo.


Doses of capecitabine and oral vinorelbine are not relevant for efficacy in breast cancer patients: an analysis of dose intensity.
The combination of capecitabine and vinorelbine is a valuable regimen in metastatic breast cancer treatment, even in pretreated patients. Forty-one pretreated consecutive patients were treated with capecitabine, 1000 mg/m2, twice daily, for two of three weeks, and vinorelbine, given orally at a dose of 60 mg/m2, days 1 and 8 in three-week cycles. A total of 301 courses was given, with a median of 8 courses (range, 3-13). Median dose intensity of capecitabine was 75% of the planned dose and for vinorelbine it was 72%. We observed 18 partial response (43.9%), 15 stable disease (36.6%), and 8 progressive disease (19.5%). Median progression-free survival was 9 months (range, 1-22) and median overall survival was 27.2 months (range, 4-40). Overall response rate (complete + partial response) was not statistically different between patients who received more or less than the median dose intensity of capecitabine and vinorelbine, and there was no difference in overall survival or progression-free survival. CONCLUSIONS; Capecitabine and oral vinorelbine is an effective and well-tolerated "all-oral" regimen for advanced breast cancer patients. The use of lower doses than those currently recommended should be not detrimental in terms of efficacy.


RNase-L-dependent destabilization of interferon-induced mRNAs. A role for the 2-5A system in attenuation of the interferon response.
The 2-5A system is an interferon-regulated RNA degradation pathway with antiviral, growth-inhibitory, and pro-apoptotic activities. RNase-L mediates the antiviral activity through the degradation of viral RNAs, and the anticellular effects of the 2-5A system are thought to be similarly mediated through the degradation of cellular transcripts. However, specific RNase-L-regulated cellular RNAs have not been identified. To isolate candidate RNase-L substrates, differential display was used to identify mRNAs that exhibited increased expression in RNase-L-deficient N1E-115 cells as compared with RNase-L-transfected cells. A novel interferon-stimulated gene encoding a 43-kDa ubiquitin-specific protease, designated ISG43, was identified in this screen. ISG43 expression is induced by interferon and negatively regulated by RNase-L. ISG43 induction is a primary response to interferon treatment and requires a functional JAK/STAT signaling pathway. The kinetics of ISG43 induction were identical in wild type and RNase-L knock-out fibroblasts; however, the decline in ISG43 mRNA following interferon treatment was markedly attenuated in RNase-L knock-out fibroblasts. The delayed shut-off kinetics of ISG43 mRNA corresponded to an increase in its half-life in RNase-L-deficient cells. ISG15 mRNA also displayed RNase-L-dependent regulation. These findings identify a novel role for the 2-5A system in the attenuation of the interferon response.


Adverse reactions to gadolinium contrast media: a review of 36 cases.
We investigated the frequency, manifestations, and severity of reactions to gadolinium contrast media in patients who underwent MR imaging at a single institution. We reviewed the quality assurance records of 21,000 patients to whom gadolinium contrast media were administered at our institution between January 1990 and October 1994. Of these patients, 36 had adverse reactions. All 36 patients were evaluated by a radiologist at the time of the adverse reaction, and a written report of the findings was completed immediately. We classified adverse reactions into four groups: mild nonallergic reactions (15 patients with nausea or vomiting), mild reactions resembling allergy (12 patients with hives, diffuse erythema, or skin irritation), moderate reactions resembling allergy (seven patients with respiratory symptoms), and life-threatening reactions resembling allergy (two patients with severe chest tightness, respiratory distress, and periorbital edema). Eleven of these 36 patients who had adverse reactions also had an unusual local reaction of skin discomfort in the extremity through which gadolinium was injected. Four of the 36 patients with adverse reactions had histories of adverse reactions to iodinated contrast media. All four of these patients and one of the remaining 32 patients with adverse reactions required treatment. No patients with adverse reactions died. Although gadolinium contrast media are safe, we found that patients had adverse reactions at a frequency greater than we expected. Severe anaphylactoid reactions occurred in two patients (0.01%). This rate exceeds the rate of 0.0003% reported in the literature. The indexes of suspicion for the occurrence of reactions to gadolinium, and both the documentation and the management of adverse reactions, must be as rigorous for reactions associated with MR imaging contrast agents as they are for reactions associated with iodinated contrast media.


Improved Doppler detection of proximal left anterior descending coronary artery stenosis after intravenous injection of a lung-crossing contrast agent: a transesophageal Doppler echocardiographic study.
This study was designed to verify the usefulness of transesophageal Doppler recording of blood flow velocity in the proximal left anterior descending coronary artery, after a peripheral injection of a lung-crossing contrast agent (SHU 508A), in detecting and locating a hemodynamically significant stenosis (vessel narrowing > or = 50%) affecting this portion of the vessel. Transesophageal Doppler echocardiography has a limited diagnostic impact on the evaluation of proximal left anterior descending coronary artery stenoses. Peripheral injection of SHU 508A, a lung-crossing contrast agent enhancing Doppler signal to noise ratio in coronary arteries, may allow recording of localized disturbed blood flow velocity at the stenosis site even in the absence of a clear B-mode visualization of the vessel. Transesophageal Doppler echocardiography, before and after echo contrast injection, was performed in 31 patients who underwent coronary angiography. Using color Doppler as a guide, pulsed wave Doppler recording of blood flow velocity in the left anterior descending coronary artery was attempted to detect a localized increase in blood flow velocity. B-mode evaluation of the vessel was also performed. Angiography showed a significant proximal left anterior descending coronary artery stenosis in 16 patients (group 1) and no stenosis in 15 patients (group 2). In 15 of 16 group 1 patients, Doppler after contrast injection revealed a localized velocity increase of at least 50% of the reference value; mean (+/-SD) percent increase in velocity was 150 +/- 89% (range 367% to 0%). In group 2 Doppler after contrast injection revealed a mild localized increase in velocity in four patients and no increase in velocity in the remaining 11 patients; mean (+/-SD) percent increase in velocity was 5 +/- 7% (range 21% to 0%, p < 0.001 vs. percent increase in group 1). When a percent velocity increase > or = 50% of the reference value was considered a positive criterion for detecting significant stenosis, the sensitivity and specificity were 92% and 100% respectively. The sensitivity of the evaluation before contrast injection or considering B-mode imaging alone was much lower (25% and 19%, respectively, p < 0.001 vs. evaluation after contrast injection). In addition, color Doppler after contrast injection correctly located the stenosis along the vessel, as compared with angiography. Blood flow evaluation of the proximal left anterior descending coronary artery by transesophageal Doppler echocardiography after contrast injection is a feasible and reliable method for detecting and locating significant stenoses affecting this part of the vessel and is an improvement over the traditional ultrasound approach.


Retinoic acid signaling in perioptic mesenchyme represses Wnt signaling via induction of Pitx2 and Dkk2.
Morphogenesis during eye development requires retinoic acid (RA) receptors plus RA-synthesizing enzymes, and loss of RA signaling leads to ocular disorders associated with loss of Pitx2 expression in perioptic mesenchyme. Several Wnt signaling components are expressed in ocular tissues during eye development including Dkk2, encoding an inhibitor of Wnt/beta-catenin signaling, which was previously shown to be induced by Pitx2 in the perioptic mesenchyme. Here, we investigated potential cross-talk between RA and Wnt signaling during ocular development. Genetic studies using Raldh1/Raldh3 double null mice deficient for ocular RA synthesis demonstrated that Pitx2 and Dkk2 were both down-regulated in perioptic mesenchyme. Chromatin immunoprecipitation and gel mobility shift studies demonstrated the existence of a DR5 RA response element upstream of Pitx2 that binds all three RA receptors in embryonic eye. Axin2, an endogenous readout of Wnt/beta-catenin signaling, was up-regulated in cornea and perioptic mesenchyme of RA deficient embryos. Also, expression of Wnt5a was expanded in perioptic mesenchyme of RA deficient eyes. Our findings demonstrate excessive activation of Wnt signaling in the perioptic mesenchyme of RA deficient mice which may be responsible for abnormal development leading to defective optic cup, cornea, and eyelid morphogenesis.


Shelf life of packaged bakery goods--a review.
Packaging requirements for fresh bakery goods are often minimal as many of the products are for immediate consumption. However, packaging can be an important factor in extending the shelf life of other cereal-based goods (toast, frozen products, biscuits, cakes, pastas). Some amount of the texture changes and flavor loss manifest over the shelf life of a soft-baked good can usually be minimized or delayed by effective use of packaging materials. The gains in the extension of shelf life will be application specific. It is recognized that defining the shelf life of a food is a difficult task and is an area of intense research for food product development scientists (food technologists, microbiologists, packaging experts). Proper application of chemical kinetic principles to food quality loss allows for efficiently designing appropriate shelf-life tests and maximizing the useful information that can be obtained from the resulting data. In the development of any new food product including reformulating, change of packaging, or storage/distribution condition (to penetrate into a new market), one important aspect is the knowledge of shelf life.


Extended Transverse-oblique Back Flap for Myelomeningocele Defect Closure: A Case Series of 10 Patients.
Myelomeningocele is the most common phenotype of congenital neural tube defects. Various reconstructive techniques have been described for soft tissue coverage following myelomeningocele repair, one of which is the use of dorsal intercostal artery perforator based flaps. The aim of this study was to describe our experience with the use of a transverse-oblique back flap that can be reliably extended to the anterior axillary line for closure of myelomeningocele defect. This pedicle transposition flap is based on thoracic or lumbar paraspinal perforators that originate from the dorsal intercostal arteries. This is a retrospective two center case-series where all patients who underwent myelomeningocele defect closure with extended transverse-oblique flap over three years period were included. Patients' clinical data, surgical variables, and outcomes were documented and analyzed using descriptive measures. Flap harvest technique is also delineated in the present study. Ten newborns (7 baby girls and 3 baby boys) who underwent a surgical closure of the myelomeningocele defect with an extended transverse-oblique back flap during the first week of life were included in the analysis. The defect was most commonly located in the lumbosacral area (50%) followed by the lumbar area (40%), with an overall average surface area of 22 ± 8.32 cm2. Common encountered complications include venous congestion to the distal part of the flap and minor wound dehiscence of less than 0.5 cm, all were managed conservatively. There were no incidences of flap loss or full necrosis. Primary closure of the donor site was possible in all cases. In the current series, the extended transverse-oblique back flap provided a safe and reliable coverage for myelomeningocele defect. Such a flap does not jeopardize other regional fasciocutanous or musclocutaneous flaps that might be needed for soft tissue coverage later in life for this population. It also obviates the placement of skin suture line over the repaired neural tube, thus yielding a durable coverage with no major complications or functional disability.


The effect of calcium channel blockers on alpha 1- and alpha 2-adrenoceptor-mediated vascular responsiveness in man.
This study investigated the effects in normotensive males of the calcium channel blockers, verapamil and nisoldipine, on the pressor responses mediated via alpha-adrenergic and non-adrenergic mechanisms. After the first doses of each drug and after 4 days continued treatment both verapamil and nisoldipine significantly attenuated the responses to angiotensin II with three- to fivefold rightward shifts of the pressor dose-response curves. Rightward shifts of comparable magnitude were obtained for phenylephrine (selective alpha 1-agonist), but with alpha-methylnoradrenaline (preferential alpha 2-agonist) only nisoldipine caused a significant twofold rightward shift and only during continued treatment. This study demonstrated that peripheral vascular adrenergic responses mediated via both alpha 1- and alpha 2-adrenoceptors in man are affected by calcium channel blocking drugs. There was no evidence that this effect was specifically linked to the alpha 2-adrenoceptor. The functional antagonism of peripheral vasopressor mechanisms was not significantly different for verapamil and nisoldipine.


Prognostic value and immunological role of AXL gene in clear cell renal cell carcinoma associated with identifying LncRNA/RBP/AXL mRNA networks.
This article aimed to explore the prognostic and immunological roles of AXL gene in clear cell renal cell carcinoma (ccRCC) for overall survival (OS) and to identify the LncRNA/RBP/AXL mRNA networks. AXL-related gene expression matrix and clinical data were obtained from The Cancer Genome Atlas (TCGA) dataset and AXL-related pathways were identified by gene set enrichment analysis (GSEA). We performed univariate/multivariate Cox regression analysis to evaluate independent prognostic factors and the relationships between AXL and immunity were also investigated. The outcomes of us indicated that the AXL mRNA expression was up-regulated in ccRCC samples and high expression of AXL was associated with worse OS in TCGA dataset (P < 0.01). Further external verification results from HPA, UALCAN, ICGC dataset, GSE6344, GSE14994, and qRT-PCR remained consistent (all P < 0.05). AXL was also identified as an independent prognostic factor for ccRCC by univariate/multivariate Cox regression analysis (both P < 0.05). A nomogram including AXL expression and clinicopathological factors was established by us and GSEA results found that elevated AXL expression was associated with the JAK-STAT, P53, WNT, VEGF and MAPK signaling pathways. In terms of immunity, AXL was dramatically linked to tumor microenvironment, immune cells, immune infiltration, immune checkpoint molecules and tumor mutational burden (TMB). As for its potential mechanisms, we also identified several LncRNA/RBP/AXL mRNA axes. AXL was revealed to play prognostic and immunological roles in ccRCC and LncRNA/RBP/AXL mRNA axes were also identified by us for its potential mechanisms.


Rotational and translational swimming of human spermatozoa: a dynamic laser light scattering study.
The rotational swimming motion of human spermatozoa is evaluated from measurements of depolarized dynamic laser light scattering at zero angle. The analysis is based on a Maxwellian angular velocity distribution and yields a rotational frequency of about 4 Hz that is ascribed to the rotation of the sperm head. From comparison with the translational swimming motion, a propelling efficiency of about 10 micron per turn is deduced. This parameter describes the linkage between the rotational and translational swimming motion and is likely to be discriminatory in the analysis of physiological and pathological sperm motions.


Complications of CSF Shunts in Pediatrics: Functional Assessment With CSF Shunt Scintigraphy-Performance and Interpretation.
OBJECTIVE. The purpose of this article is to review the performance method and criteria for interpretation of CSF shunt scintigraphy studies. CONCLUSION. Interpretation of CSF shunt scintigraphy studies requires an in-depth understanding of hydrocephalus, the functioning of CSF shunts and their components, and the mechanisms of failure of such devices. Application of strict interpretive criteria when evaluating CSF shunt scintigraphy studies improves diagnostic yield, providing valuable functional information to the neurosurgical team who manages patients with shunted hydrocephalus.


Reconstruction of the neck with two rotation-advancement platysma myocutaneous flaps.
A reconstruction of a neck with a defect caused by radionecrosis sequelae using two rotation-advancement platysma myocutaneous flaps is presented. The thinness of the flaps, their accessibility, the lack of bulk, and the primary closure of the donor site, without functional or aesthetic problems, all render this technique an attractive option for replacing anterior neck skin.


Neurocognitive, clinical and functional correlates of subjective quality of life in Asian outpatients with schizophrenia.
Quality of life (QOL) impairment is evident in patients with schizophrenia and is increasingly recognised as an important evaluation criterion of treatment outcome. Hence, this study aimed to identify the neurocognitive, clinical and functional parameters associated with subjective QOL in patients with schizophrenia within an Asian context, and specifically in an outpatient setting. This study was conducted on 83 outpatients with DSM-IV diagnosis of schizophrenia, and 47 age- and gender-matched healthy controls. All participants were administered with the World Health Organisation Quality of Life Assessment-Brief Form (WHOQOL-BREF) and Brief Assessment of Cognition in Schizophrenia (BACS), to measure quality of life and cognitive function respectively. Patients were also assessed for severity of psychopathology, as well as level of psychosocial functioning, using the Positive and Negative Syndrome Scale (PANSS) and Global Assessment of Functioning (GAF) rating scales respectively. Specific psychopathology (greater severity of PANSS negative symptoms, general psychopathology subscale scores), cognitive deficits (working and verbal memories), and lower GAF scores were correlated with poorer QOL in patients. Multivariate analyses revealed that younger age, being single and lower level of psychosocial functioning were associated with poorer QOL but level of psychosocial functioning did not appear to mediate the effects of symptoms and neurocognitive deficits on QOL. Overall, this study highlighted the need for clinicians to pay more attention to these clinical, neurocognitive and functional parameters and their integrative relationships with QOL in order to optimise the treatment outcomes of patients with schizophrenia.


Reduced airway responsiveness in nonelite runners.
The effects of endurance training on airway responsiveness in nonasthmatic subjects are poorly defined. We hypothesized that airway responsiveness may differ between none-lite endurance athletes and sedentary subjects, and studied healthy, nonelite runners and sedentary controls by single-dose methacholine challenges carried out in the absence of deep inspirations, in that deep inspirations are known to oppose airway narrowing in nonasthmatic subjects. A total of 20 nonasthmatic none-lite runners (mean age+/- SD: 43.0+/- 8.5 yr; training volume: 68 km.wk; range: 40-100; racing experience: 11+/- 8 yr) and 20 sedentary controls (age: 44.0+/- 20.6 yr) were studied, all of them being normo-reactive to standard methacholine challenge up to 25 mg.mL concentration. All subjects were studied at rest; six runners were also studied about 1 h after completing the Palermo marathon (December 8, 2001). The primary outcome of the study was the inspiratory vital capacity (IVC) obtained after single-dose methacholine inhalation at the end of 20 min of deep inspiration prohibition. At rest, IVC decreased by 10.5+/-8.1% after challenge with methacholine at 75 mg.mL in athletes, and by 24.3+/-16.1% after a methacholine concentration of 52+/-5.7 mg.mL in sedentary controls (P=0.002). The decreased response to methacholine in runners did not correlate with static lung volumes, amount of weekly training, or running experience. Methacholine challenge under deep inspiration prohibition revealed that endurance training attenuates airway responsiveness in nonasthmatic, none-lite runners. Airway hyporesponsiveness was potentiated after the marathon, suggesting involvement of humoral (i.e., catecholamine levels), airway factors (i.e., nitric oxide), or both in modulating airway tone after exercise.


Increased prion protein processing and expression of metabotropic glutamate receptor 1 in a mouse model of Alzheimer's disease.
Prion protein (PrP(C) ), a glycosylphosphatidylinositol-anchored protein corrupted in prion diseases, has been shown recently to interact with group I metabotropic glutamate receptors (mGluRs). Moreover, both PrP(C) and mGluRs were proposed to function as putative receptors for β-amyloid in Alzheimer's disease. PrP(C) can be processed in neurons via α or β-cleavage to produce PrP(C) fragments that are neuroprotective or toxic, respectively. We found PrP(C) α-cleavage to be 2-3 times higher in the cortex of APPswe/PS1dE9 mice, a mouse model of Alzheimer's disease. A similar age-dependent increase was observed for PrP(C) β-cleavage. Moreover, we observed considerable age-dependent increase in cortical expression of mGluR1, but not mGluR5. Exposure of cortical neuronal cultures to β-amyloid oligomers upregulated mGluR1 and PrP(C) α-cleavage, while activation of group I mGluRs increased PrP(C) shedding from the membrane, likely due to increased levels of a disintegrin and metalloprotease10, a key disintegrin for PrP(C) shedding. Interestingly, a similar increase in a disintegrin and metalloprotease10 was detected in the cortex of 9-month-old APPswe/PS1dE9 animals. Our experiments reveal novel and complex processing of PrP(C) in connection with mGluR overexpression that seems to be triggered by β-amyloid peptides. Prion protein (PrP(C) ) and metabotropic glutamate receptors (mGluR) are implicated in Alzheimer's disease (AD). We found age-dependent increase in PrP(C) processing, ADAM10 and mGluR1 levels in AD mouse model. These changes could be reproduced in cultured cortical neurons treated with Aβ peptide. Our findings suggest that increased levels of Aβ can trigger compensatory responses that may affect neuronal toxicity.


Evaluation of anti-inflammatory and antinociceptive activity of Triphala recipe.
The anti-inflammatory and antinociceptive activities of Triphala recipe were studied in animal models. Triphala recipe (4 mg/ear) significantly exhibited an inhibitory effect on the ear edema formation induced by ethyl phenylpropiolate-induced, but not on the arachidonic acid-induced ear edema in rats. Furthermore, Triphala recipe at the doses of 300, 600 and 1,200 mg/kg significantly reduced carrageenan-induced hind paw edema. Next, the anti-inflammatory action in chronic inflammation was measured using the cotton pellet-induced granuloma formation assay in rats. Triphala recipe (1,200 mg/kg) reduced neither transudative weight nor granuloma formation. It also did not affect on body weight gain and thymus weight indicating that Triphala recipe does not have a steroid-like effect. In antinociceptive study, Triphala recipe (300, 600, 1,200 mg/kg), elicited significant inhibitory effect on both phases, especially in late phase, of the formalin test in mice suggesting that the antinociceptive action of Triphala recipe may be via both peripheral and at least partly centrally acting.


Prevalence of hearing loss among Canadians aged 20 to 79: Audiometric results from the 2012/2013 Canadian Health Measures Survey.
In Canada, population-level estimates of hearing loss have been based on self-reported data, yielding estimates of 4% or 5%. Self-reported hearing difficulties may result in underestimates of hearing loss, particularly among people with mild loss and among older adults. The 2012/2013 Canadian Health Measures Survey (cycle 3) collected audiometric and self-reported data to estimate the prevalence of hearing loss and limitations in a population-based sample of Canadians aged 20 to 79. Weighted frequencies and cross-tabulations were used to calculate measured and self-reported hearing levels by sociodemographic characteristics. All estimates were weighted at the person-level to represent the population. Based on a pure-tone average of four frequencies that are important in speech, 19.2% of Canadians aged 20 to 79 had measured hearing loss in at least one ear; 35.4% had high-frequency hearing loss. These levels exceeded the self-reported estimate of hearing difficulty--3.7%--derived from responses to questions from the Health Utilities Index Mark 3. The prevalence of measured hearing loss rose with age from no more than 10% among people younger than 50 to 65% at ages 70 to 79. Men were more likely than women to have a hearing loss, a difference that emerged around age 60. Canadians with low household income and/or educational attainment were more likely than those in higher income/education households to have a hearing loss. This analysis presents the first population-based audiometric data on the prevalence of hearing loss among the adult household population of Canada, and highlights the disparity between measured and self-reported outcomes.


Omalizumab: when the non-responder is a late-responder.
Omalizumab is an anti-IgE monoclonal antibody available since 2006 for the treatment of GINA step 4 asthma. We studied a 41-year old male who has been suffering from severe steroid-resistant asthma with severe co-morbidity and treated with Omalizumab. He was found to be non-responder to the treatment until the 48th week, starting from which we began to see a distinct improvement in the symptoms and all the correlated parameters, in addition to remission of the co-existent allergy to milk. we wish to point out the late response to Omalizumab, which occurred way beyond the times envisaged in literature. It seems possible that some patients are late responders to the drug.


Errors in force generation and changes in controlling patterns following agonist muscle fatigue.
The purpose of this study was to evaluate whether agonist muscle fatigue changed the coactivation time and the co-contraction magnitude of the agonist and antagonist muscle, and if the agonist muscle fatigue produced bias (constant error: CE) and inconsistency (variable error: VE) of the force. Subjects are 10 healthy people and one person with impaired proprioception. EMG and force for fast (0.19 +/- 0.06 s) and slow (1.20 +/- 0.44 s) targeted isometric dorsiflexions were recorded before and after fatigue of the dorsiflexors. The results revealed that the coactivation time increased after fatigue only in the slow contractions but the co-contraction magnitude did not change. The postfatigue increment of the CE was greater in the fast contractions than in the slow ones. We conclude that the postfatigue compensatory strategy can reduce the fatigue-induced bias. The change of muscles activation level after fatigue might be under the influence of the common drive. Impaired proprioception is a possible cause of the fatigue-related increase in bias and inconsistency.


Calcium channel ITPR2 and mitochondria-ER contacts promote cellular senescence and aging.
Cellular senescence is induced by stresses and results in a stable proliferation arrest accompanied by a pro-inflammatory secretome. Senescent cells accumulate during aging, promoting various age-related pathologies and limiting lifespan. The endoplasmic reticulum (ER) inositol 1,4,5-trisphosphate receptor, type 2 (ITPR2) calcium-release channel and calcium fluxes from the ER to the mitochondria are drivers of senescence in human cells. Here we show that Itpr2 knockout (KO) mice display improved aging such as increased lifespan, a better response to metabolic stress, less immunosenescence, as well as less liver steatosis and fibrosis. Cellular senescence, which is known to promote these alterations, is decreased in Itpr2 KO mice and Itpr2 KO embryo-derived cells. Interestingly, ablation of ITPR2 in vivo and in vitro decreases the number of contacts between the mitochondria and the ER and their forced contacts induce premature senescence. These findings shed light on the role of contacts and facilitated exchanges between the ER and the mitochondria through ITPR2 in regulating senescence and aging.


[Minimal invasiveness of laparoscopic radical prostatectomy: reality or dream?].
Currently, radical retropubic prostatectomy is the standard procedure for clinically localized prostate cancer. The surgical technique has been continuously refined for decades, resulting in reduced morbidity and improved functional and oncologic results. Since the late 90s, radical prostatectomy has been increasingly performed laparoscopically. A search of the available data has found that the articles published so far have proven the feasibility of the laparoscopic procedure but never confirmed its less invasiveness. In accordance with previous studies that have evaluated the invasiveness of various open and laparoscopic procedures, our clinic, which has routinely performed both techniques for several years, addressed the question whether laparoscopic prostatovesiculectomy indeed induces less severe surgical trauma. This prospective nonrandomized comparison study of the University Clinic of Urology at the Martin-Luther University at Halle-Wittenberg recruited a total of 64 patients, who underwent laparoscopic radical prostatectomy (n = 32) or open retropubic prostatectomy (n = 32) from January 2003 to April 2004. Both patient groups were comparable as to preoperative staging, PSA value and Gleason score. Besides perioperative parameters, such as surgical time, intra- and postoperative complications, blood loss and transfusion rate, need for analgetics and length of hospital stay, the comparison included oncologic data, such as Gleason score, pathologic stage and numbers of positive specimen margins. To get objectively reproducible data, the range of the systemic answers concerning the surgically induced tissue trauma was recorded as laboratory data. In all patients, pre-, intra-, and postsurgical markers of the acute-phase C-reactive protein, serum amyloid A (SAA), interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured. The transfusion rate was 6 % for laparoscopic prostatectomies and 12 % for open prostatectomies. A rectal tear had to be intraoperatively repaired in one laparoscopically operated patient. The postoperative use of analgetics was comparable in both groups. The median hospital stay was 12.4 days for the laparoscopic and 11.2 days for the open surgical group. For T2 tumors, positive specimen margins were found in 6 cases (17 %) of the laparoscopic and in 4 cases (12 %) of the open surgical group. As to the indicators of any systemic reaction, no significant difference could be found during the entire clinical course between both surgical methods. In comparison with patients who underwent conventional open prostatectomy, patients with laparoscopically radical prostatectomy had identical to slightly higher serum levels of the acute-phase parameters, as evidence of an equal or a discretely manifested systemic response to the surgical trauma. The so far assumed less invasiveness of laparoscopic radical prostatectomy is not objectively supported by the data from this study. Thus, surgical trauma and its linked invasiveness must be considered equal for both methods, at least for the time being.


Central motor conduction time in malnourished children.
The functional status of the descending motor pathways was assessed in malnourished children using transcranial electromagnetic stimulation of the cortex. Twenty children with different severities of malnutrition and 20 control subjects were studied electrophysiologically. The circular coil of a Dentac MAG 2 magnetic stimulator was applied tangentially over the vertex to stimulate the cortex. The muscle evoked potential in the children's arms was recorded from the abductor pollicis brevis muscle and in their legs from the extensor digitorum brevis muscle on both sides of the body using surface electrodes. The muscle evoked potential of the abductor pollicis brevis and extensor digitorum brevis muscles was further obtained using root stimulation by applying the coil at the cervical and lumbar spine, respectively. The indices of cortical threshold, cortical latency, and central motor conduction time (ms) were evaluated in both arm and leg muscles on both sides. The results showed an increased cortical threshold (mean (SD) 1232.5 (134.06) in the study group v 1147.5 (99.31) in the control group) for the abductor pollicis brevis muscle and for the extensor digitorum brevis muscle (1325.00 (115.70) in the study group v 1190.0 (125.24) in the control group). Similarly, significant prolongation of the central motor conduction time (ms) (study group 6.67 (0.91) v control group 5.71 (0.74)) in the abductor pollicis brevis muscle was seen in malnourished children.


Histologic and morphologic character of pediatric abdominal aortic developmental coarctation and hypoplasia.
Abdominal aortic coarctation and hypoplasia are uncommon diseases, recognized most often in pediatric-aged individuals. Comprehensive studies regarding the pathologic spectrum of these aortopathies are nonexistent. This investigation was undertaken to better define the histologic and morphologic character of abdominal aortic narrowings affecting children and assess its potential relevance to contemporary clinical practice. Aortic specimens obtained during open operations in children being treated for symptomatic, noninflammatory abdominal aortic narrowings at the University of Michigan were subjected to histologic study after hematoxylin and eosin, Movat, Verhoeff Van Gieson, and Masson's trichrome preparations. Microscopic findings were correlated with the anatomic aortic images. In addition, a detailed review was completed of all prior reports in the English literature that included images depicting the histologic character of noninflammatory abdominal aortic narrowings in children. Among a series of 67 pediatric-aged individuals undergoing open surgical interventions for abdominal aortic narrowings, eight children ranging in age from 9 months to 18 years, had adequate aortic tissue available for study. The loci of the specimens paralleled the anatomic sites of segmental coarctations observed in the entire series, with involvement of the suprarenal abdominal aorta (n = 3), intrarenal aorta (n = 2), and infrarenal aorta (n = 1). Diffusely hypoplastic abdominal aortas (n = 2) included one case of a de facto aortic duplication, represented by a channel that paralleled the narrow native aorta and gave origin to celiac artery branches, as well as the superior mesenteric and renal arteries. Concentric or eccentric intimal fibroplasia was observed in every aorta, often with internal elastic fragmentation and duplication (n = 4). Media abnormalities included elastic tissue disorganization (n = 3) and focal medial fibrosis (n = 1). Organizing luminal thrombus occurred in two infants. Coexistent ostial stenoses of the celiac, superior mesenteric, or renal arteries were observed in all but the only child who had an infrarenal aortic coarctation. Neurofibromatosis type 1 affected one child whose histologic findings were indistinguishable from those of the other children. A review of prior published histologic images of abdominal aortic coarctation and hypoplasia affecting children from other centers revealed a total of 14 separate reports, each limited to single case photomicrographs, of which 11 exhibited intimal fibroplasia. Intimal fibroplasia is a common accompaniment of developmental abdominal aortic coarctation and hypoplasia. It is posited that intimal fibroplasia, which is likely progressive in instances of abnormal shear stresses in these diminutive vessels, may contribute to less salutary outcomes after endovascular and certain open reconstructions of pediatric abdominal aortic narrowings.


[Histopathology and ultrastructural changes of the musculature and nerves during methamidophos toxication in chicken].
After exposure to methamidophos (Via digestive or respiratory tract or just by skin contamination) for 1, 2, 3, 4, 8 and 12 weeks respectively, ultrastructure of the myofibrils in chicken with methamidophos toxication via the digestive tract showed mild degeneration in the first week and serious degeneration by the end of the fourth week. Light microscopy of the muscle-fibril showed atrophy and necrosis in the third week, including both myopathic atrophy and neuropathic atrophy. Recovery of the lesions took place from the 8th to the 12th week (Light or elcetion microscopy). Those chickenstaking immunosuppressive drug simultaneously with methamidophos exposure (via digestive tract) showed less degeneration of the myofibrils and nerves. Ultrastructure of the myofibril in chickens with methamidophos intoxication through the respiratory tract also developed less degeneration of myofibrils and nerves than those obtained through digestive intoxication. There was no lesion obtained in chicken taking methamidophos via the respiratory tract coexistently with the immunosuppressive drug or just by skin contamination.


The COPE-48: An adapted version of the COPE inventory for use in clinical settings.
The Coping Orientation to Problems Experienced inventory (COPE) is one of the most frequently used tools for assessing coping styles. However, there is a lack of information about its psychometric properties in applied settings. The aim of this study is to analyze the psychometric properties and internal structure of the COPE in an adult clinical sample attending a primary care mental health unit and to develop an adapted version to increase its reliability and clinical utility. We administered the questionnaire to 301 participants (37.5% men) aged 18-69 years. The most prevalent principal diagnoses were adjustment (39.9%) and mood disorders (35.9%). The principal component analysis yielded a nine factor solution accounting for 61.23% of the total variance. This shortened version of the COPE provides a theoretically and empirical consistent inventory, improves the factor structure and increases the reliability of the scales with a mean alpha value of .81. This version, the COPE-48, offers good psychometric properties, it takes about 20min to administer, allowing a reliable and parsimonious evaluation. Its administration in applied settings could help with identifying which coping strategies are used and which can be modified and promoted by psychological treatment.


The effects of amosulalol, a novel combined alpha- and beta-adrenoreceptor antagonist, on the rat isolated right ventricle and aorta.
The effect of amosulalol on the accumulation of radioactivity from [3H]-noradrenaline and on the subsequent spontaneous and nerve stimulation-evoked outflow of radioactivity have been investigated in the rat isolated right ventricle. In addition the effect of amosulalol on the contractions of the electrically-driven directly muscle stimulated rat right ventricle to isoprenaline and of the rat isolated aorta to phenylephrine and 5-hydroxytryptamine are reported. Amosulalol at 10(-6)M did not prevent the accumulation of radioactivity from a solution containing [3H]-noradrenaline. The spontaneous outflow of radioactivity, following loading of the ventricle with [3H]-noradrenaline, was increased by amosulalol at 10(-6)M by a cocaine- and idazoxan- insensitive mechanism. The nerve stimulation-evoked outflow of radioactivity was increased by amosulalol (10(-6)M), cocaine (10(-5)M) and idazoxan (10(-7)M). The ability of amosulalol to increase nerve-evoked outflow was maintained in the presence of cocaine but prevented by pretreatment with idazoxan. This suggests that amusulalol is an alpha 2-adrenoreceptor antagonist. The contractile response of the electrically-driven directly muscle stimulated right ventricle to isoprenaline were inhibited by amosulalol at 10(-7) and 10(-6)M with apparent pA2 values of 7.5 and 8.1, respectively. It is suggested that amosulalol at 10(-6)M may have an action additional to beta 1-adrenoreceptor antagonism on the right ventricle. The contractile responses of the rat aorta to phenylephrine were inhibited by amosulalol at 10(-7) and 10(-6)M with a pA2 of 8.6 which was independent of concentration. Amosulalol also reduced the magnitude of the maximal responses of the aorta to 5-hydroxytryptamine.


Ultra-Weak Chemiluminescence Enhanced by Cerium-Doped LaF3 Nanoparticles: A Potential Nitrite Analysis Method.
In this work, cerium-doped LaF3 nanoparticles (LaF3:Ce NPs) were successfully synthesized and characterized. Its chemiluminescence (CL) property was studied, and it was amazingly found that it intensely enhanced the ultra-weak CL of the NaNO2-H2O2 system. The CL mechanism was systematically investigated and suggested to be the recombination of electron-injected and hole-injected LaF3:Ce NPs. The new CL system was developed to be a facile, original, and direct method for nitrite analysis. Experimental conditions were optimized and then a satisfactory linear relationship between CL intensity and nitrite concentration was obtained. This work introduced a new pathway for the research and application of traditional fluoride NPs doped with RE3+.


Efficacy of Germicidal Hand Wash Agents in Use in a Meat Processing Plant.
The in-use efficacy of a selected range of germicidal hand wash agents was tested in a meat processing plant. The hand washes included non-germicidal soaps and germicidal agents containing chlorhexidine, iodophor and Irgasan DP 300 as active ingredients. A laboratory study was done under controlled conditions with standardized procedures for hand washing; in the meat plant, "normal" (unstandardized) hand wash procedures were followed. Levels of contamination on hands varied markedly between work units. Only in the meat cutting area could a significant difference be attributed to hand wash agents against transient-type bacteria on workers' hands. The hand wash agent with 4% chlorhexidine gluconate, the iodophor with 0.75% available iodine and the gel containing 0.3% Irgasan DP 300 were the only products that gave a significantly better reduction of transient bacteria than non-germicidal soap. Transient bacteria were detected on hands after washing, indicating that under the in-use conditions in the meat processing plant, hand wash techniques did not remove all of these bacteria from hands. The plant workers generally indicated a dislike for the iodophor products as hand germicides.


[Para-esophageal hernias. Apropos of 42 cases].
Between 1979 and May 1994, 42 patients (11 males, 31 females, mean age 72.8 +/- 12.4 years) underwent surgery for paraoesophageal hiatal hernia. In 37 cases, clinical signs had been observed for 64 months on the average: digestive (32), respiratory (15), cardiac (9). Nine patients had iron deficiency anaemia. Eighteen patients had a complication (acute obstruction: 10, haemorrhage: 6, perforation: 2, lung infection: 2). In 5 cases, the complication was the inaugural sign. Surgery was performed in a emergency situation in 5 cases and deferred emergency in 12 and surgery was elective in 25. An antireflux procedure was used in 36 cases (Lortat-Jacob (2), Toupet (5), Dor (2), Nissen-Rossetti (27)). Another procedure was also needed in 12 patients. There were no postoperative deaths and complications occurred in 6 patients (14.9%). Outcome was known in 40 patients with a mean follow-up of 40.5 months (range 3 to 140 months). Functional results was very good in 35 patients (87.5%), good in 3 (7.5%) and poor in 2 (5%). The most satisfactory immediate results were obtained for patients who had elective surgery and in those with a deferred emergency. Use of the Nissen-Rossetti procedure did not have any deleterious effect on these results and no long-term inconveniences.


Raloxifene, estrogen, and alendronate affect the processes of fracture repair differently in ovariectomized rats.
We investigated the effects of inhibitors of bone resorption (estrogen, raloxifene, and alendronate) on the processes of fracture repair in ovariectomized (OVX) rats. One hundred forty female Sprague-Dawley rats at 3 months of age were either OVX or sham-operated and divided into five groups: sham control, OVX control, estrogen (17alpha-ethynyl estradiol [EE2], 0.1 mg/kg), raloxifene (Rlx, 1.0 mg/kg), and alendronate (Aln, 0.01 mg/kg) groups. Treatment began immediately after the surgery. Four weeks postovariectomy, prefracture controls were killed and bilateral osteotomies were performed on the femoral midshafts and fixed with intramedullary wires. Treatment was continued and fracture calluses were excised at 6 weeks and 16 weeks postfracture for evaluation by X-ray radiography, quantitative computed tomography (QCT,) biomechanical testing, and histomorphometry. At 6 weeks postfracture, Aln and OVX had larger calluses than other groups. Sham and OVX had higher ultimate load than EE2 and Rlx, with Aln not different from either control. Aln calluses also contained more mineral (bone mineral content [BMC]) than all other groups. By 16 weeks postfracture, OVX calluses were smaller than at 6 weeks and the dimensions for Aln had not changed. Aln had higher BMC and ultimate load than OVX, EE2, and Rlx. EE2 and Rlx had similar biomechanical properties, which were similar to sham. Interestingly, OVX and Aln animals were heavier than other groups at all time points; therefore, ultimate load was normalized by body weight to show no significant differences in strength of the whole callus between groups at either 6 weeks or 16 weeks postfracture. However, Aln strongly suppressed remodeling of the callus, resulting in the highest content of woven bone, persistent visibility of the original fracture line, and lowest content of lamellar bone, compared with other groups. Therefore, the larger Aln callus appeared to be a remarkable, morphological adaptation to secure the fracture with inferior material. In conclusion, OVX-stimulated bone turnover resulted in the fastest progression of fracture repair that was most delayed with Aln treatment, consistent with marked suppression of bone resorption and formation activity. Estrogen and Rlx had similar effects that were generally similar to sham, indicating that mild suppression of bone turnover with these agents has insignificant effects on the progression of fracture repair.


Misidentification of transthyretin and immunoglobulin variants by proteomics due to methyl lysine formation in formalin-fixed paraffin-embedded amyloid tissue.
Proteomics is becoming the de facto gold standard for identifying amyloid proteins and is now used routinely in a number of centres. The technique is compound class independent and offers the added ability to identify variant and modified proteins. We re-examined proteomics results from a number of formalin-fixed paraffin-embedded amyloid samples, which were positive for transthyretin (TTR) by immunohistochemistry and proteomics, using the UniProt human protein database modified to include TTR variants. The amyloidogenic variant, V122I TTR, was incorrectly identified in 26/27 wild-type and non-V122I variant samples due to its close mass spectral similarity with the methyl lysine-modified WT peptide [126KMe]105-127 (p.[146 KMe]125-147) generated during formalin fixation. Similarly, the methyl lysine peptide, [50KMe]43-59, from immunoglobulin lambda light chain constant region was also misidentified as arising from a rare myeloma-derived lambda variant V49I. These processing-derived modifications are not present in fresh cardiac tissue, non-fixed fat nor serum and do not materially affect the identification of amyloid proteins. They could result in the incorrect assignment of a variant, and this may have consequences for the immediate family who will require genetic counselling and potentially early clinical intervention. As proteomics becomes a routine clinical test for amyloidosis, it becomes important to be aware of potentially confounding issues such as formalin-mediated lysine methylation, and how these may influence diagnosis and possibly treatment.


Retinoic acid induces stage-specific repatterning of the rostral central nervous system.
We had previously reported that in gastrulating mouse embryos retinoic acid (RA) induces morphological as well molecular alterations strictly depending on the time of administration. In particular, embryos treated with RA at the mid-late streak stage share reduction of the rostral central nervous system (CNS) and increase of the hindbrain. In the same embryos, loss of the forebrain-expressed genes, such as Emx1, Emx2, and Dlx1, and rostral ectopic expression of the Hoxb-1 gene suggest an antero-posterior (A/P) ordered repatterning of the fore-, mid-, and hindbrain regions. Several genes, such as Pax-2, Wnt-1, En-2, and En-1, are involved in the establishment of midbrain and rostral hindbrain regional identities and boundaries. We report that these genes become coordinately anteriorized only in embryos treated with RA at the late streak stage. Moreover, in the hindbrain of the same embryos, at 8.5 days post coitum (dpc), Wnt-1 and Pax-2 are rostrally induced all along the neural plate. Considering that forebrain markers are repressed in embryos treated with RA at the same time, these findings strongly support the idea that RA administration at the late streak stage induces an ordered repatterning of the rostral CNS, possibly altering the A/P nature of mesendodermal inductive signals.