Sequence analysis of the Porphyromonas gingivalis dipeptidyl peptidase IV gene.
We previously constructed a Porphyromonas gingivalis genomic library and isolated the 2.9 kb EcoRV fragment which specified glycylprolyl dipeptidyl aminopeptidase (GPase). Nucleotide sequencing of this fragment identified the single 2169 bp open reading frame which coded for a 723 amino acid protein. The amino acid sequencing of the NH2-terminal domain of the native and recombinant mature enzymes suggested that the protease possessed a 16 amino acid residue signal peptide. The calculated mass of the precursor and mature proteases were 82,018 and 80,235 daltons, respectively. The homology search of this enzyme in registered protein sequences revealed that this enzyme was homologous to dipeptidyl peptidase (DPP) IV from the Flavobacterium meningosepticum and that from eukaryotic cells, including the human, mouse, and rat. Three amino acid residues, Ser-593, Asp-668, and His-700, were identified as a putative catalytic triad, a common feature of eukaryotic serine proteases. In addition, this enzyme showed a broad proteolytic spectrum toward synthetic substrates capable of splitting not only Gly-Pro-derivative but also Ala-Pro, Lys-Pro, and Phe-Pro-derivatives. Therefore, we conclude that this enzyme belongs to DPP IV rather than GPase.


[Surgical therapy of non-Hodgkin lymphoma of the stomach].
The value of surgical treatment is less well defined in gastric lymphoma than in gastric carcinoma. Therefore, we analysed the outcome of 245 patients with gastric malignancies, which were treated from 1.1.1988 to 31.12.1995 in the Department of Surgery/Charité. Twenty patients suffered from a non-Hodgkin-lymphoma and only 14 (8%) were diagnosed correctly by preoperative endoscopical biopsy. Seven patients with limited lymphoma underwent primary surgical therapy (total gastrectomy n = 4, subtotal gastrectomy n = 3). Seven patients with disseminated lymphoma (stage EIII-IV) were treated by neoadjuvant chemotherapy (CHOP), that was followed by gastrectomy (R0: n = 2, R1: n = 4) or explorative laparotomy (n = 1). Five patients (25%) were diagnosed as undifferentiated carcinoma and underwent total gastrectomy with D2-lymphadenectomy (R0: n = 5). One patient (5%) underwent emergency surgery due to gastric perforation. One patient (5%) died in hospital due to insufficient anastomosis after total gastrectomy and preoperative chemotherapy. Nine patients with high-grade malignant lymphomas received postoperative chemotherapy. The 1-, 2,- and 5-year-survival-rate was 95%, 89% and 44%. Although, many questions are still open, surgical therapy remains an important option in the multimodal treatment of gastric lymphoma.


Effect of insulin on plasma norepinephrine and 3,4-dihydroxyphenylalanine in obese men.
Increasing evidence relates serum insulin level and blood pressure in obese individuals. Although the connection between these two factors is not established, a common presumption is that the sympathetic nervous system is somehow involved, in part, because laboratory studies demonstrate insulin stimulation of sympathetic and cardiovascular activity. Because the obese may exhibit altered responsiveness to insulin action, the current investigation compared cardiovascular and neurohumoral responses to euglycemic insulin infusion (200 mU/m2/min) in obese and lean men. At baseline, obese men displayed higher glucose and insulin levels, faster pulse rates, and elevated mean arterial pressures (MAP) than lean controls; plasma norepinephrine (NE) and 3,4-dihydroxyphenylalanine (DOPA) concentrations, however, did not differ. During insulin infusion, pulse rate increased equally in obese and lean subjects (from 69 to 78 min-1 in obese and from 56 to 66 min-1 in lean subjects), while MAP remained unchanged in both groups. Elevations in plasma NE (+85 pg/mL in obese and +109 in lean pg/mL) and reductions in plasma DOPA (-233 pg/mL in obese and -376 pg/mL in lean) did not differ between groups. Sodium excretory rate decreased during insulin infusion in lean subjects by 2.2 mEq/h but increased in obese by 5.3 mEq/h (difference in response between groups, P = .024). Thus, in these obese men, cardiovascular and sympathetic responses to insulin persist despite evidence of moderate insulin resistance; increased sympathetic activity, as a cause for the resting tachycardia and borderline hypertension at baseline, seems unlikely.


Biomolecular logic gate for analysis of the New Delhi metallo-β-lactamase (NDM)-coding gene with concurrent determination of its drug resistance-encoding fragments.
We herein exploit a newly schemed logic gate to superiorly facilitate analysis of long and highly structured nucleic acids. This strategy uniquely enables the identification of NDM-specific genes and concurrent screening of two active site-encoded fragments, which is promising for evaluating microbial drug resistance.


The antioxidant EPC-K1 attenuates NO-induced mitochondrial dysfunction, lipid peroxidation and apoptosis in cerebellar granule cells.
In this study we investigated the effects of nitric oxide (NO) on cultured cerebellar granule cells. Exposure to NO donors, S-nitrosoglutathione (GSNO; 250 microM) or sodium nitroprusside (SNP; 500 microM), triggered apoptosis in immature cultures of cerebellar granule cells, which was characterized by chromatin condensation, nuclei fragmentation, and DNA laddering. Exposure of cerebellar granule cells to NO donors led to a decrease in the mitochondrial transmembrane potential and intracellular ATP content, which suggested that NO treatment caused mitochondrial dysfunction. NO treatment also induced oxidative stress in cerebellar granule cells as measured by thiobarbituric acid (TBA) assay. Pretreating cells with L-ascorbic acid 2-[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H -1-benzopyran-6-yl-hydrogen phosphate] potassium salt (EPC-K1), a novel antioxidant, attenuated NO-induced mitochondrial dysfunction and oxidative stress to some extent, and prevented the cells from apoptosis. The results of the present investigation suggest that a superoxide/peroxynitrite-mediated oxidative stress may be an important pathway leading to NO-associated neuronal damage. Pretreating cells with the antioxidant EPC-K1 attenuated NO-induced neurotoxicity by scavenging superoxide/peroxynitrite and/or its breakdown products.


The genome as pharmacopeia: association of genetic dose with phenotypic response.
The majority of compounds that enter clinical trials fail to meet regulatory standards for marketed therapeutics. Success in clinical trials is increased when therapies are directed against targets for which genetic variants have been linked to disease risk or progression in humans. Here we review how genome-wide association studies (GWAS) have furthered our understanding of the genetic basis of common disease, and how GWAS findings could stimulate development of novel drugs. Genetic screens that associate variants with clinical phenotypes of interest, like pharmacological screens that associate compounds with in vitro responses of interest, establish an intervention-response relationship on which a therapeutic hypothesis can be based. We highlight the similarities between the GWAS approach and well-established principles in early drug discovery, from primary screen through lead identification. We believe that integration of genetic and pharmacological screens will considerably enhance the development of therapeutic interventions with clear benefit to patients.


Transplantable thyroid tumour in rats: iodoamino acid distribution in successive tumour generations.
All first generation thyroid tumours in rats, produced by 3 micronc 131I followed by LID, were uniformly functional containing MIT, DIT, T4, T3, and in some cases, T'3. All fourth generation tumours were nonfunctional as tested. Second and third generation tumours were biochemically unpredictable, some containing all IAA including T'3, others only MIT, I-and a trace DIT. Degree of dedifferentiation and of autonomy did not necessarily correlate with loss of function. Results could be explained by postulating nodules with differing biochemical properties within individual tumours.


Genetically predicted testosterone and cardiovascular risk factors in men: a Mendelian randomization analysis in the Guangzhou Biobank Cohort Study.
Observationally lower testosterone is associated with an unhealthier cardiovascular (CVD) risk profile, but this association is open to confounding and reverse causality. The authors examined the association of testosterone with well-established cardiovascular disease (CVD) risk factors (blood pressure, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL)cholesterol and fasting glucose) and the Framingham score using a Mendelian randomization analysis with a separate-sample instrumental variable estimator. To minimize reverse causality, a genetic score predicting testosterone was developed in 289 young Chinese men from Hong Kong, based on three selected testosterone-related single nucleotide polymorphisms (rs10046, rs1008805 and rs1256031). Multivariable censored and linear regressions were used to examine the association of genetically predicted testosterone levels with CVD risk factors and Framingham score among 4212 older Chinese men from the Guangzhou Biobank Cohort Study. Predicted testosterone was unrelated to systolic blood pressure [-0.11 mmHg, 95% confidence interval (CI) -0.70 to 0.48], diastolic blood pressure (0.04 mmHg, 95% CI -0.27 to 0.36), fasting glucose (0.02 mmol/l, 95% CI -0.02 to 0.06) or Framingham score (0.02, 95% CI -0.0001 to 0.03) but associated with higher LDL-cholesterol (0.02 mmol/l, 95% CI 0.01 to 0.04) and lower HDL-cholesterol (-0.01 mmol/l, 95% CI -0.02 to -0.001), after adjustment for potential confounders (age, education, smoking status, use of alcohol and body mass index). Our findings did not corroborate observed protective effects of testosterone on cardiovascular risk factors or risk of ischaemic heart disease among men, but raises the possibility that higher testosterone may be associated with an unhealthier lipid profile.


Single-nucleotide polymorphism array karyotyping in clinical practice: where, when, and how?
Single-nucleotide polymorphism array (SNP-A) karyotyping is a new technology that has enabled genome-wide detection of genetic lesions in human cancers, including hematopoietic neoplasms. Taking advantage of very large numbers of allele-specific probes synthesized on microarrays at high density, copy number alterations as well as allelic imbalances can be sensitively detected in a genome-wide manner at unprecedented resolutions. Most importantly, SNP-A karyotyping represents the only platform currently available for genome-scale detection of copy neutral loss of heterozygosity (CN-LOH) or uniparental disomy (UPD), which is widely observed in cancer genomes. Although not applicable to detection of balanced translocations, which are commonly found in hematopoietic malignancies, SNP-A karyotyping technology complements and even outperforms conventional metaphase karyotyping, potentially allowing for more accurate genetic diagnosis of hematopoietic neoplasms in clinical practice. Here, we review the current status of SNP-A karyotyping and its application to hematopoietic neoplasms.


Efficacy of Sofosbuvir, Velpatasvir, and GS-9857 in Patients With Hepatitis C Virus Genotype 2, 3, 4, or 6 Infections in an Open-Label, Phase 2 Trial.
Studies are needed to determine the optimal regimen for patients with chronic hepatitis C virus (HCV) genotype 2, 3, 4, or 6 infections whose prior course of antiviral therapy has failed, and the feasibility of shortening treatment duration. We performed a phase 2 study to determine the efficacy and safety of the combination of the nucleotide polymerase inhibitor sofosbuvir, the NS5A inhibitor velpatasvir, and the NS3/4A protease inhibitor GS-9857 in these patients. We performed a multicenter, open-label trial at 32 sites in the United States and 2 sites in New Zealand from March 3, 2015 to April 27, 2015. Our study included 128 treatment-naïve and treatment-experienced patients (1 with HCV genotype 1b; 33 with HCV genotype 2; 74 with HCV genotype 3; 17 with genotype HCV 4; and 3 with HCV genotype 6), with or without compensated cirrhosis. All patients received sofosbuvir-velpatasvir (400 mg/100 mg fixed-dose combination tablet) and GS-9857 (100 mg) once daily for 6-12 weeks. The primary end point was sustained virologic response 12 weeks after treatment (SVR12). After 6 weeks of treatment, SVR12s were achieved by 88% of treatment-naïve patients without cirrhosis (29 of 33; 95% confidence interval, 72%-97%). After 8 weeks of treatment, SVR12s were achieved by 93% of treatment-naïve patients with cirrhosis (28 of 30; 95% CI, 78%-99%). After 12 weeks of treatment, SVR12s were achieved by all treatment-experienced patients without cirrhosis (36 of 36; 95% CI, 90%-100%) and 97% of treatment-experienced patients with cirrhosis (28 of 29; 95% CI, 82%-100%). The most common adverse events were headache, diarrhea, fatigue, and nausea. Three patients (1%) discontinued treatment due to adverse events. In a phase 2 open-label trial, we found sofosbuvir-velpatasvir plus GS-9857 (8 weeks in treatment-naïve patients or 12 weeks in treatment-experienced patients) to be safe and effective for patients with HCV genotype 2, 3, 4, or 6 infections, with or without compensated cirrhosis. ClinicalTrials.gov ID: NCT02378961.


On traditional dental extraction: case reports from Dinka and Nuer en route to restoration.
Ritual dental extraction among Sub-Saharan African populations has been practised for centuries, yet little is known about the removal process for any ethnic group. Dinka and Nuer refugees to the US requested replacements for missing anterior teeth removed during childhood. Among 36 Sudanese refugees, 238 individual extractions had been performed. Three retained canine/incisor root fragments; their cases are presented, including memories of the tooth-extraction ritual.


High-resolution ultrasound imaging to demonstrate and predict efficacy of carbon dioxide fractional resurfacing laser treatment.
Fractional resurfacing is a new laser treatment used mainly for treating photoaging and acne scars. To demonstrate the efficacy of carbon dioxide (CO(2) ) fractional resurfacing laser treatment using high-resolution ultrasound imaging and to evaluate factors predictive of response to the treatment. A prospective observational study was undertaken from January 2007 to August 2009. Laser treatment was performed using a CO(2) fractional resurfacing device. Patients were offered high-resolution ultrasound imaging to follow up their treatment. Dermal and subepidermal nonechogenic band (SENEB) thicknesses were measured. Twenty-four consecutive patients were included, allowing follow-up of 107 areas. Statistical analysis showed a significant increase in dermal thickness (mean variation +25.1%, (95% confidence interval=19.0-31.2). SENEB thickness was not changed. Older age and greater initial thickness were associated with a smaller increase in dermal thickness according to multivariate analysis. The region treated was also found to be a significant predictive factor. Smoking, topical anesthesia, severity of local side effects induced by the laser treatment, and phototype were not associated with significant modification of dermal thickness after laser treatment. This is the first report on the use of high-resolution ultrasound imaging to assess the efficacy of fractional laser operating with standard parameters. Increase in dermal thickness was evidenced, possibly explained by collagen neosynthesis or conformational changes in the extracellular matrix component, increasing dermal hydration, although there was no modification of SENEB thickness, suggesting that the skin retains the sequelae of ageing. The treatment appeared to be more effective in younger patients, probably because their fibroblasts are more prone to synthesizing collagen. Thinner skins were more effectively treated than thicker skins because they could be stimulated throughout their thickness with the treatment parameters used.


Cerebrospinal fluid acetylcholinesterase changes after treatment with donepezil in patients with Alzheimer's disease.
We analyzed whether donepezil differently influences acetylcholinesterase (AChE) variants from cerebrospinal fluid (CSF) in patients with Alzheimer's disease (AD) after long-term treatment. Overall CSF-AChE activity in AD patients before treatment was not different from controls, but the ratio between the major tetrameric form, G(4), and the smaller G(1) and G(2) species was significantly lower. AChE levels at study outset were found to correlate positively with beta-amyloid (1-42) (Abeta42). When patients were re-examined after 12 months treatment with donepezil, there was a remarkable increase in both the G(4) and the lighter species of CSF AChE. As compared with placebo, donepezil caused decreases in the percentage of AChE that failed to bind to the lectin concanavalin A and the antibody AE1. These non-binding species comprised primarily a small subset of G(1) and G(2) forms. In treated patients, these light variants were the only subset of CSF AChE that correlated with CSF-Abeta42 levels. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that a 77-kDa band, attributed in part to inactive AChE, was lower in AD patients than in controls. Unlike enzyme activity, the intensity of this band did not increase after donepezil treatment. The varying responses of different AChE species to ChE-I treatment suggest different modes of regulation, which may have therapeutic implications.


Distribution patterns of Wolbachia endosymbionts in the closely related flower bugs of the genus Orius: implications for coevolution and horizontal transfer.
Vertical transmission is the primary route of the endosymbiont Wolbachia for its own spread among invertebrate hosts, but horizontal transmission between different hosts is believed to have occurred multiple times. However, it is not well known how Wolbachia commonly spread among closely related hosts. We focused on the closely related species of the minute pirate bugs belonging to the genus Orius, which are important biological control agents in agricultural crops because they are the most useful natural enemy of various tiny pests, such as thrips. Here, we examined five Orius species (Orius sauteri, Orius nagaii, Orius minutus, Orius strigicollis, and Orius tantillus) from eight geographic localities in Japan for Wolbachia infection. Two distinct strains, wOus1 and wOus2, were detected based on Wolbachia surface protein (wsp) gene sequencing. Furthermore, multilocus sequence typing revealed that each of the strains comprised two variants that differed in a single nucleotide. The overall distribution patterns of the two Wolbachia strains were found to differ among host species: prevalent double infection with wOus1 and wOus2 in O. strigicollis; fixation of single infection with wOus2 in O. nagaii; occurrence of single infection with wOus1 in O. sauteri; prevalence of single infection with wOus1 in O. minutus with an exception in a single population; and lack of Wolbachia infection in O. tantillus. Such differences in the distribution patterns of Wolbachia may reflect the evolutionary history of Wolbachia infection among Orius species and/or ecological and physiological differences among the Orius species that determine the invasiveness and maintenance of the two Wolbachia strains.


Effect of the hydrophobic basal layer of thermoresponsive block co-polymer brushes on thermally-induced cell sheet harvest.
Thermoresponsive poly(benzyl methacrylate)-b-poly(N-isopropylacrylamide) (PBzMA-b-PIPAAm) block co-polymer brush surfaces were prepared by surface-initiated two-step reversible addition-fragmentation chain transfer radical (RAFT) polymerization. PBzMA brushes were fabricated on azoinitiator-immobilized glass substrates in the presence of dithiobenzoate (DTB) compound as a RAFT agent. The amount of grafted polymer was regulated by initial monomer concentrations. The second thermoresponsive blocks were added to the RAFT-related DTB groups located at PBzMA termini through the propagation of PIPAAm chains, resulting in formation of PBzMA-b-PIPAAm brushes. Surface characteristics of the block co-polymer brushes and its influence on thermally regulated cellular behavior were investigated using bovine carotid artery endothelial cells (BAECs), compared with PIPAAm brush surfaces. Cell adhesion/detachment behavior on thermoresponsive polymer brush surfaces significantly depended on their individual polymer architectures and chemical compositions of grafted polymers. Low-temperature treatment at 20°C, below the phase-transition temperature of PIPAAm, induced the spontaneous detachment of adhering cells from the PBzMA-b-PIPAAm brush surfaces with a higher rate than that from PIPAAm brush surfaces. In addition, the cell-repellent effect of the hydrophobic basal layer successfully accelerated for harvesting BAEC sheets from the block co-polymer brush surfaces. Unique features of thermoresponsive block co-polymer brush architectures can be applied to control cell-adhesion strength for enhancing cell adhesion or accelerating cell detachment.


An expanded phylogeny of treefrogs (Hylidae) based on nuclear and mitochondrial sequence data.
The treefrogs (Hylidae) make up one of the most species-rich families of amphibians. With 885 species currently described, they contain >13% of all amphibian species. In recent years, there has been considerable progress in resolving hylid phylogeny. However, the most comprehensive phylogeny to date (Wiens et al., 2006) included only 292 species, was based only on parsimony, provided only poor support for most higher-level relationships, and conflicted with previous hypotheses in several parts (including the monophyly and relationships of major clades of Hylinae). Here, we present an expanded phylogeny for hylid frogs, including data for 362 hylid taxa for up to 11 genes (4 mitochondrial, 7 nuclear), including 70 additional taxa and >270 sequences not included in the previously most comprehensive analysis. The new tree from maximum likelihood analysis is more well-resolved, strongly supported, and concordant with previous hypotheses, and provides a framework for future systematic, biogeographic, ecological, and evolutionary studies.


Dietary orotic acid accentuates the hepatic response to phenobarbital in rats.
In rats treated with phenobarbital for 3 days and simultaneously fed a semisynthetic diet containing 1.0% orotic acid, the extent of the increases in liver microsomal phosphatidylcholine, phosphatidylethanolamine, total RNA, total protein, and cytochrome P-450 were significantly greater than they were in rats treated identically with phenobarbital but without dietary orotic acid. This is attributed primarily to the stimulation of hepatic phosphatidylcholine synthesis by dietary orotic acid. In the absence of phenobarbital, orotic acid was shown to cause some increase in liver smooth endoplasmic reticulum components, but not cytochrome P-450. Orotic acid also decreased the activity of microsomal phosphatidylethanolamine N-methyltransferase, which may have contributed to the increase in the microsomal content of phosphatidylethanolamine. The hypothesis is advanced that phospholipid availability is a limiting factor in the hepatic response to phenobarbital. When more phospholipid is available to provide the structural framework for biogenesis of endoplasmic reticulum, all of the hepatic actions of phenobarbital, including induction of cytochrome P-450, are amplified.


Cyclosporin A inhibition of aggrecanase-mediated proteoglycan catabolism in articular cartilage.
To determine the effect of cyclosporin A (CSA) on aggrecanase- and matrix metalloproteinase (MMP)-mediated catabolism of proteoglycan (aggrecan) in articular cartilage explants stimulated with interleukin-1 (IL-1) in a culture system that mimics early pathologic processes associated with arthritic disease. Proteoglycan (glycosaminoglycan) and lactate quantification, Western immunoblot analyses of aggrecan degradation products, reverse transcription-polymerase chain reaction analyses of aggrecanase-1, aggrecanase-2 (ADAM-TS4, ADAM-TS5, respectively), MMP-1, MMP-3, MMP-13, tissue inhibitor of metalloproteinases 1 (TIMP-1), TIMP-2, and TIMP-3 messenger RNA (mRNA) expression in articular cartilage explant cultures, and electrophoretic mobility shift assay analysis of nuclear factor of activated T cells (NF-AT) transcription factor activation were used. CSA inhibited, in a dose-dependent and noncytotoxic manner, aggrecanase-mediated proteoglycan catabolism and loss from IL-1-stimulated cartilage explants. There was no evidence of MMP-mediated aggrecan catabolism in this in vitro model. Treatment of articular cartilage explant cultures with 10 ng/ml of IL-1alpha up-regulated the expression of mRNA for ADAM-TS4, ADAM-TS5, MMP-1, MMP-3, and MMP-13. The expression of ADAM-TS4, ADAM-TS5, and MMP-13 was abrogated by the inclusion of 10 microM CSA in the culture medium. NF-AT activation was observed in chondrocytes but could not be inhibited by preincubation with 10 microM CSA. CSA can inhibit IL-1-induced aggrecanase-mediated proteoglycan catabolism in articular cartilage explants maintained in culture for 4 days, thus demonstrating molecular mechanisms whereby CSA may be an effective therapy for degenerative joint disease.


Fluoro-edenite fibres induce lung cell apoptosis: an in vivo study.
We previously showed that apoptosis in the lungs of sheep exposed to fluoro-edenite fibres is induced via the receptor pathway. The present study was performed to gain further insights into the mechanisms of activation of programmed cell death induced by the fibres. Fluoro-edenite fibres are similar in size and morphology to some amphibolic asbestos fibres. They have been found in benmoreitic lavas, in the local stone quarry, in building materials and in road paving at Biancavilla, a town in eastern Sicily (Italy), where epidemiological surveys revealed a cluster of mortality from pleural mesothelioma. Inhalation of asbestos fibres can cause chronic inflammation and carcinogenesis. Since fluoro-edenite has been shown to activate the apoptotic process, we set out to characterise the expression of apoptosis-regulating proteins in fluoro-edenite-exposed lung disease and sought to determine if apoptosis results from fluoro-edenite exposure. Lung tissue from apparently healthy sheep habitually grazing near Biancavilla was processed for immunohistochemical localisation of bcl-2 and bax. Results showed epithelial and interstitial bax overexpression, especially in cells directly in contact with the fibres, and negative bcl-2 immunoexpression. TUNEL-positive cells were detected in alveoli and connective tissue. The integrity of alveolar epithelium and alveolar apoptosis are critical determinants in the pathways that initiate fibrogenesis in the lung and fibroblastic foci are usually found close to abnormal or denuded alveolar epithelium. Our results are consistent with the hypothesis that apoptosis is an important mechanism for removing cells with irreparable fluoro-edenite-induced genetic changes that predispose them to a neoplastic evolution.


Prognosis of airway obstruction and feeding difficulty in the Robin sequence.
To evaluate the course and prognosis of airway obstruction and feeding difficulty in the Pierre Robin sequence (PRS). Retrospective review of 60 patients with PRS between 1993 and 2002 at the University of California, Davis Medical Center. Patients were placed into diagnostic subgroups: (1) Isolated PRS; (2) Syndromic PRS (known syndrome with PRS); (3) Unique PRS (unique anomalies with PRS). Data regarding severity, duration, and management of airway obstruction and feeding difficulty were collected. Airway obstruction requiring intervention beyond positional therapy was seen in 28% isolated, 42% syndromic, and 58% unique PRS. One-third of patients who failed positional therapy were temporarily stabilized with a nasopharyngeal airway or endotracheal intubation. The remaining two-thirds of patients, who failed positional therapy required a surgical airway procedure. Four patients underwent mandibular distraction osteogenesis, resulting in successful decannulation or avoidance of tracheostomy. Thirteen patients underwent tracheostomy; mean duration of tracheostomy-dependence was 17.0 months in Isolated PRS and 31.7 months in Unique PRS (p < 0.01). Successful decannulation by age of 3 years was confirmed in 85% of patients who underwent tracheostomy. Tube feeding was required in 53% Isolated, 67% Syndromic, and 83% Unique PRS. Forty-two percent of PRS patients with a successful positional airway still demonstrated feeding difficulty. Short-term (0-3 months) and intermediate (4-18 months) tube feeding was more commonly required in Isolated and Syndromic PRS, while long-term (beyond 18 months) gastrostomy tube feeding was more commonly required in Unique PRS (p < 0.01). By 3 years of age, a successful oral diet was seen in 91% Isolated, 92% Syndromic, and 78% Unique PRS. Diagnostic subgroups based on the presence of additional anomalies help families and physicians in understanding the severity and duration of feeding and airway difficulty in PRS. Two-thirds of PRS patients who fail positional therapy may ultimately require a surgical airway procedure. Feeding difficulty can be present in the absence of clinically significant airway obstruction. Families and physicians should be encouraged that by 3 years of age, most patients were successfully taking an oral diet without airway obstruction.


Prospective evaluation of carotid artery stenosis: elliptic centric contrast-enhanced MR angiography and spiral CT angiography compared with digital subtraction angiography.
Although digital subtraction angiography (DSA) is the reference standard for assessing carotid arteries, it is uncomfortable for patients and has a small risk of disabling stroke and death. These problems have fueled the use of spiral CT angiography and MR angiography. We prospectively compared elliptic centric contrast-enhanced MR angiography and spiral CT angiography with conventional DSA for detecting carotid artery stenosis. Eighty carotid arteries (in 40 symptomatic patients) were assessed. Elliptic centric MR and spiral CT angiographic data were reconstructed with maximum intensity projection and multiplanar reconstruction techniques. All patients had been referred for DSA evaluation on the basis of findings at Doppler sonography, which served as a screening method (degree of stenosis > or = 70% or inconclusive results). Degree of carotid stenosis estimated by using the three modalities was compared. Significant correlation with DSA was found for stenosis degree for both elliptic centric MR and spiral CT angiography; however, the correlation coefficient was higher for MR than for CT angiography (r = 0.98 vs r = 0.86). Underestimation of stenoses of 70-99% occurred in one case with elliptic centric MR angiography (a 70% stenosis was underestimated as 65%) and in nine cases with spiral CT angiography, in comparison to DSA findings. Overestimation occurred in two cases with MR angiography (stenoses of 65-67% were overestimated as 70-75%). With CT, overestimation occurred in seven cases; a stenosis of 60% in one case was overestimated as 70%. Both techniques confirmed the three cases of carotid occlusion. With elliptic centric MR angiography, carotid stenoses of 70% or greater were detected with high sensitivity, 97.1%; specificity, 95.2%; likelihood ratio (LR) for a positive test result, 20.4; and ratio of LR(+) to LR(-), -0.3. With spiral CT angiography, sensitivity, specificity, LR(+), and LR(+):LR(-) were 74.3%, 97.6%, 31.2, and 0.3, respectively. Elliptic centric contrast-enhanced MR angiography is more accurate than spiral CT angiography to adequately evaluate carotid stenosis. Furthermore, elliptic centric contrast-enhanced MR angiography appears to be adequate to replace conventional DSA in most patients examined.


Environmental and biological monitoring of occupational exposure to polynuclear aromatic hydrocarbons during highway pavement construction in Italy.
We performed a cross-sectional study with the main aim of evaluating occupational exposure to polycyclic aromatic hydrocarbons (PAHs) in workers involved in the pavement construction of a new highway in Northern Italy, where modified bitumen was used as binder for Hot Mix Asphalt. We applied a combined approach of air and biological monitoring. Both the aerosol and vapour phases of bitumen were collected applying the NIOSH 5506 method. The 16 PAHs listed as high priority by EPA were determined by HPLC-UV. End-of-shift urine samples were collected from 144 workers to determine 1-hydroxypyrene (1-OHP) and 2-naphthol (2-NAP) concentrations after enzyme digestion and HPLC-UV analysis. Socio-demographic and lifestyle information was collected by a questionnaire. Paving workers were actually exposed to PAHs, including carcinogenic compounds, that were measurable only in the aerosol phase. Higher exposure as well as dose levels were measured for the paver group. Biological monitoring confirmed that 1-OHP was less affected by smoking habits as compared to 2-NAP and showed a higher association with occupational exposure. Carcinogenic PAH compounds were detectable only in the aerosol phase and this must be taken into account in the adoption of preventive measures. Biomonitoring supported the superiority of 1-OHP as compared to 2-NAP in assessing the internal dose in such workers.


Quantitative imaging of apoptosis commitment in colorectal tumor cells.
We have studied caspase-3 activation by combined DNA damage induction and EGFR kinase inhibition in order to identify potential EGFR-mediated survival signals conferring resistance to apoptosis in human colorectal tumor cells. The onset of apoptosis was microscopically imaged with a newly developed caspase-3 substrate sensor based on EGFP and tHcred1, enabling us to monitor caspase-3 activation in cells by fluorescence lifetime imaging microscopy or fluorescence correlation spectroscopy. Both optical approaches provide parameters quantitatively reporting the ratio between cleaved and uncleaved sensor, thereby facilitating the comparison of caspase-3 activation between different cells. Using these methods, we show that EGFR kinase inhibitors sensitize colorectal SW-480 tumor cells for 5-fluorouracil-induced apoptosis, indicating that EGFR-mediated survival signaling contributes to apoptosis resistance via its intrinsic kinase activity.


Effects of highly active antiretroviral therapy on vaccine-induced humoral immunity in HIV-infected adults.
The acquisition of adequate vaccine-induced humoral immunity is especially important in HIV-infected individuals, who are at increased risk of infections. The aim of the study was to assess the safety of administering a complete vaccination programme to successfully treated HIV-infected adults and to evaluate specific humoral responses and the effect of highly active antiretroviral therapy (HAART) interruption on these responses. A placebo-controlled, double-blind clinical trial was designed and 26 HIV-infected adults enrolled. Study participants were randomized to receive either a complete immunization schedule with commercial vaccines or placebo for 12 months. HAART was then discontinued for 6 months. Specific humoral responses were evaluated at baseline, at month 12 and after HAART interruption and compared between groups. There were neither local nor systemic secondary effects related to vaccination. Specific humoral responses to vaccines were adequate, but a loss of immunoglobulin G titres was observed after HAART interruption in 12 study participants. HAART interruption may cause impairment of previously acquired vaccine-induced immunity in HIV-infected adults.


Sexual and reproductive health in Greenland: evaluation of implementing sexual peer-to-peer education in Greenland (the SexInuk project).
Background For decades, the rates of sexually transmitted infections (STIs), such as gonorrhoea, chlamydia and syphilis, have increased in Greenland, especially within the young age groups (15-29 years). From 2006 to 2013, the number of abortions has been consistent with approximately 800-900 abortions per year in Greenland, which is nearly as high as the total number of births during the same period. Previous studies in Greenland have reported that knowledge about sexual health is important, both as prevention and as facilitator to stop the increasing rates of STIs. A peer-to-peer education programme about sexual health requires adaption to cultural values and acceptance among the population and government in order to be sustainable. Objective Formative evaluation of a voluntary project (SexInuk), in relation to peer-to-peer education with focus on sexual health. Two workshops were conducted in Nuuk, Greenland, to recruit Greenlandic students. Design Qualitative design with focus group interviews (FGIs) to collect qualitative feedback on feasibility and implementation of the project. Supplemented with a brief questionnaire regarding personal information (gender, age, education) and questions about the educational elements in the SexInuk project. Eight Greenlandic students, who had completed one or two workshops, were enrolled. Results The FGIs showed an overall consensus regarding the need for improving sexual health education in Greenland. The participants requested more voluntary educators, to secure sustainability. The articulation of taboo topics in the Greenlandic society appeared very important. The participants suggested more awareness by promoting the project. Conclusion Cultural values and language directions were important elements in the FGIs. To our knowledge, voluntary work regarding peer-to-peer education and sexual health has not been structurally evaluated in Greenland before. To achieve sustainability, the project needs educators and financial support. Further research is needed to investigate how peer-to-peer education can improve sexual and reproductive health in Greenland.


Relationships between metals exposure and epidemiological parameters of two pathogens in urban pigeons.
Human activities often generate or increase concentration of chemical compounds including pesticides, hydrocarbons and metals that can potentially affect ecological interactions. We found that elevated levels of zinc in pigeon feathers were associated with both low prevalence of Chlamydiaceae (ornithosis disease) and low intensity of blood pathogens (Haemosporidian parasites). In contrast, high levels of lead in pigeon feathers were associated with high blood pathogens intensities. Our results suggest that metals linked to human activities in cities such as zinc and lead may play a significant role in the ecology of host-parasite interactions and could potentially affect the epidemiology of diseases in the urban environment.


Dosimetric study on eye's exposure to wide band radio frequency electromagnetic fields: variability by the ocular axial length.
This paper evaluates the variability of specific absorption rate (SAR) in the human eye. This variability results from changes in ocular axial length (OAL), which is common in many ophthalmologic and vision abnormalities, including myopia. A generic eye model was reconstructed according to published data. The feasibility of using the generic model in numerical research of electromagnetic fields (EMF) was demonstrated by means of comparative simulations with eye models reconstructed from magnetic resonance (MR) scans. Free-form deformation (FFD) was used to deform the OAL of the generic eye model. Thus, 64 deformed eyes were created and were categorized according to the OAL increase. The finite-difference time-domain (FDTD) method was applied in the simulations. The results revealed that changing the OAL does not increase EMF absorption in the eyes or the eye tissues. No additional induced temperature rise was produced by the changes of OAL. The results also indicated that the non-pathological increment of the OAL, which is inevitable during the childhood, does not increase the SAR in the eyes.


Origin and mechanisms of non-disjunction in human autosomal trisomies.
Chromosomal aneuploidy is one of the major causes of pregnancy wastage. In this review we summarize the knowledge about the origin and mechanisms of non-disjunction in human autosomal trisomies 8, 13, 15, 16, 18, and 21, accumulated during the last decade by using DNA polymorphism analysis. Maternal meiosis I non-disjunction is the most important single class, but chromosome-specific patterns exist. For the acrocentric chromosomes 15 and 21, meiosis I errors predominate among the maternal errors, in contrast to trisomy 18 where meiosis II errors predominate. For trisomy 16, virtually all cases are due to maternal meiosis I non-disjunction. Postzygotic (mitotic) non-disjunction constitutes 5-15% of cases of trisomies 15, 18, and 21, whereas for trisomy 8 and trisomy 8 mosaicism the majority of cases are due to mitotic non-disjunction. For paternal non-disjunction of chromosomes 18 and 21, meiosis II or mitotic errors predominate. There is aberrant meiotic recombination associated with maternal meiotic non-disjunction in all trisomies studied in detail so far. Advanced maternal age remains the only well documented risk factor for maternal meiotic non-disjunction, but there is, however, still a surprising lack of understanding of the basic mechanism(s) behind the maternal age effect.


Endoglin regulates nitric oxide-dependent vasodilatation.
Endoglin is a membrane glycoprotein that plays an important role in cardiovascular development and angiogenesis. We examined the role of endoglin in the control of vascular tone by measuring nitric oxide (NO)-dependent vasodilation in haploinsufficient mice (Eng+/-) and their Eng+/+ littermates. The vasodilatory effect of acetylcholine, bradykinin, and sodium nitroprusside was assessed in anesthetized mice; in isolated, perfused hindlimbs; and in aortic rings. The substantial hypotensive and vasodilatory response induced by acetylcholine and bradykinin in Eng+/+ was markedly reduced in Eng+/- mice. Both kinds of animals had similar responses to sodium nitroprusside, suggesting that the deficient vasodilatory effect is not due to a NO response impairment. Urinary and plasma concentrations of nitrites, a NO metabolite, were lower in Eng+/- than in Eng+/+ mice. The levels of endothelial nitric oxide synthase (eNOS) in kidneys and femoral arteries were about half in Eng+/- than in Eng+/+ mice and were also reduced in primary cultures of aortic endothelial cells from Eng+/- compared with those from Eng+/+ mice. Furthermore, overexpression or suppression of endoglin in cultured cells induced a marked increase or decrease in the protein levels of eNOS, respectively. Thus, our results in vivo and in vitro demonstrate a relationship between endoglin and NO-dependent vasodilation mediated by the regulation of eNOS expression.


Galvanically induced body sway in the anterior-posterior plane.
Anterior-posterior body sway was evoked with monopolar bi-aural galvanic stimulus of the vestibular nerves in normal subjects and recorded with a force platform, two experiments being conducted. In an experiment of paired design, 9 normal subjects showed an increase in anterior-posterior sway as compared with lateral sway when exposed to the stimulus. In a second experiment another group of 10 normal subjects were exposed to a galvanic stimulus between a neck electrode and two electrodes placed on the arms, but there was no change in the relationship between anterior-posterior and lateral body sway. It is concluded that monopolar galvanic stimulus of the vestibular nerves can induce anterior-posterior body sway, a phenomenon which can be utilised to investigate the vestibulo-spinal contribution in postural control in the anterior-posterior plane.


Assessment of the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique.
To evaluate the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique. 14 dogs (similarly housed and fed identical diets). Samples of intestinal contents were collected from the duodenum, jejunum, ileum, colon, and rectum of each dog. Bacterial DNA was extracted from the samples, and the variable V6 to V8 region of 16S ribosomal DNA (gene coding for 16S ribosomal RNA) was amplified by use of universal bacterial primers; polymerase chain reaction amplicons were separated via denaturing gradient gel electrophoresis (DGGE). Similarity indices of DGGE banding patterns were used to assess variation in the bacterial microflora among different compartments of the intestine within and among dogs. Bacterial diversity was assessed by calculating the Simpson diversity index, the Shannon-Weaver diversity index, and evenness. DGGE profiles indicated marked differences in bacterial composition of intestinal compartments among dogs (range of similarity, 25.6% to 36.6%) and considerable variation among compartments within individual dogs (range of similarity, 36.7% to 579%). Similarities between neighboring intestinal compartments were significantly greater than those between non-neighboring compartments. Diversity indices for the colon and rectum were significantly higher than those of the duodenum, jejunum, and ileum. Results indicated that the different intestinal compartments of individual dogs appear to host different bacterial populations, and these compartmental populations vary among dogs. In dogs, fecal sample analysis may not yield accurate information regarding the composition of bacterial populations in compartments of the gastrointestinal tract.


Novel biodegradable polydioxanone stents in a rabbit airway model.
This study was undertaken to evaluate safety and biocompatibility of a novel biodegradable polydioxanone stent in a rabbit tracheal model. Metallic and silicone stents represent standard therapeutic approaches for hollow organ stenosis, although complications have been reported repeatedly. Biodegradable stents could reduce the risks associated with this procedure while still achieving the purpose of maintaining lumen patency. A commercially available polydioxanone suture strand with a long safety record was used to manufacture the self-expanding stents. The polydioxanone stents were then implanted bronchoscopically and under fluoroscopic guidance into the tracheas of white rabbits (N = 25). Periodic clinical examination was performed. Histopathologic examination concluded the study for the 5 experimental groups at 3, 4, 5, 10, and 15 weeks after implantation. There were no unexpected deaths and no stent displacements during the study. The animals remained in good condition, without stent debris expectoration. Macroscopic examination revealed that the tracheal lumen stayed open. Histologic examination showed that tracheal damage score was highest 5 weeks after stenting, including in-stent necrosis of the epithelium. Stent degradation was complete with no remnants after 10 weeks, leaving the trachea completely healed at 15 weeks after implantation. This animal airway model has demonstrated acceptable safety and biocompatibility of this novel biodegradable polydioxanone stent. We suggest that polydioxanone stenting be used for further clinical studies for cases in which complete stent degradation after temporary airway treatment is desirable.


Characterization of aggregates of hepatitis C virus glycoproteins.
Hepatitis C virus (HCV) encodes two glycoproteins, E1 and E2, which assemble in oligomeric structures. Studies of HCV glycoprotein assembly using heterologous expression systems have shown that these glycoproteins can follow two pathways: a productive pathway leading to the formation of a non-covalent heterodimer; and a non-productive pathway leading to the formation of large disulfide-linked aggregates. The non-covalent HCV glycoprotein complex is probably the functional complex which plays an active role in the entry process in host cells. The aggregates are believed to be waste products; however, one can imagine that, in infected cells, they could provide HCV glycoproteins with additional functions. To further understand the potential role played by HCV glycoprotein aggregates in HCV infection, a MAb (H14) was produced which specifically recognizes these aggregates but not the non-covalent E1E2 heterodimer. The H14 epitope was shown to be present on both HCV glycoproteins and was sensitive to deglycosylation. An additional characterization of HCV glycoprotein aggregates, with the help of MAb H14, indicates that they share an epitope with a cellular protein called Mac-2 binding protein. The presence of such an epitope on HCV glycoprotein aggregates could potentially lead to the production of autoantibodies recognizing Mac-2 binding protein in HCV-infected patients.


Extending the working pH of nitrobenzene degradation using ultrasonic/heterogeneous Fenton to the alkaline range via amino acid modification.
Oxides of iron, α-Fe2O3 (I), and copper, CuO (II) prepared by usual precipitation method without surfactant were used at room temperature in the process of nitrobenzene (10mgL(-1)) degradation at different pH values with ultrasonic at 20kHz. The degradation was complete in 20 and 30min for (I) and (II), respectively in the pH range 2-7 using1.0gL(-1) of solids and 10mM of H2O2. A remarkable decrease in degradation efficiency was recorded on increasing the pH to values higher than the neutral range. This loss in efficiency was cancelled to a great extent through modifying the used oxides with amino acids. Arginine showed higher improving effect to (II) (1:1 weight ration) than glycine or glutamic acid. Modification of both oxides with increasing amounts of arginine increased the degradation efficiency of (I) in a more regular way than in case of (II). However, the extent of improvement due to amino acid modification was higher in case of (II) because of its originally low degradation efficiency in strongly alkaline media.


Foley catheter, a simple device for removal of oesophageal foreign bodies.
An oesophageal foreign body is a common mishap in children. Removal through oesophagoscopy is a standard procedure but where this facility is not available this rather common accident could pose serious problems. A foley catheter was used repeatedly for the extraction of smooth foreign bodies from the oesophagus without fluoroscopic control. The procedure is safe and could be used whenever oesophagoscopy is not available.


Anatomic parameters of the sacral lamina for osteosynthesis in transverse sacral fractures.
To analyze the morphometric parameters of the dorsal sacral lamina and pedicles to determine if there is adequate bony architecture to support plate osteosynthesis. Two reviewers performed measurements on 98 randomly selected high-resolution CT scans of the pelvis to quantify the bony anatomy of the sacral lamina. Measurements included the depths of the lamina at each sacral level, the trajectory and depth of the sacral pedicles from the sacral lamina, and the width of the sacral canal. A bone mineral density analysis was performed on the sacral lamina using Hounsfield units (HU) and compared to the L1 and S1 vertebral bodies. The sacral lamina were found to form peaks and troughs which we refered to as major and minor sections. On average, the thickness was > 4 mm at all major screw starting points, indicating adequate geometry for screw fixation. The sacral pedicle depths were 27, 18, 16, and 14 mm at S2-S5, respectively. The average angulation from midline of this screw path directed laterally to avoid the sacral canal was 20°, 17°, 8°, and - 8° for the S2-5 pedicles, respectively. Average sacral canal diameter was 11 mm for S2 and 8 mm for S3-5. The sacral lamina had an average bone mineral density of 635 HU, which was significantly different from the density of the L5 (220 HU) and S1 (165 HU) vertebral bodies (p < 0.005). This morphometric data was used to pilot a new plating technique. The sacral lamina offers a novel target for screw fixation and meets the basic geometric and compositional criteria for screw purchase. To our knowledge, this study represents the first morphometric analysis performed on the sacral lamina and pedicles for plate application.


Effect of phenolic compounds on in vivo activity of glucose-6-phosphatase in certain tissues of Notopterus notopterus.
The effects of phenol (P), dinitrophenol (DNP), pentachlorophenol (PCP), and their three combinations--(PCP + DNP)/P (highly antagonistic), (DNP + P)/PCP (additive) and (P + DNP)/PCP (highly synergistic)-- on glucose-6-phosphatase activity in liver, brain, kidney and gills of Notopterus notopterus were studied at three sub-acute concentrations (1/10, 1/15 and 1/20 the fraction of the 96-h LC50) after 15 and 30 days of exposure. When these chemicals were used separately, PCP exerted more effect than DNP and P. Maximum (71.56%) and highly significant (P less than 0.001) inhibition was observed in liver at the highest concentration of (P + DNP)/PCP combination after 30 days exposure, and lowest (3.64%) and insignificant inhibition in kidney at 1/20 of the (PCP + DNP)/P combination after 15 days exposure. Significant (P less than 0.05, P less than 0.01) stimulation in kidney or no effect in other tissues was observed in lower concentrations of P, DNP and (PCP + DNP)/P.


[Morphological changes in the gastric and duodenal mucosa in duodenal ulcer in young persons].
Histological investigations were carried out upon the state of the gastric and duodena mucosa in 27 young patients with duodenal ulcer, seven out of them being in the state of exaverbation of the ulcer and 20-in remission. While the duodenal mucosa in the majority of the cases (88.9%) was established to be affected, in 85.2 percent of the patients the gastric mucosa remained intact. The chronic duodenitis observe n 24 of the cases have no atrophic character. The changes in the mucous secretion of duodenal mucosa are in direct correlation with the state of exacerbation or remission of the ulcer process. Similar correlation was not established in gastric mucosa.


Genetic dissection of gustatory sensitivity to bitterness (sucrose octaacetate) in mice.
Genes implicated in consumption of a bitter compound, sucrose octaacetate (SOA), were investigated using a full genomic scanning strategy. For a 0.1 mM concentration, two QTL reached 5.8 and 6.5 lod scores on chromosomes 2 (77 cM) and 11 (14 cM), respectively. For a 1 mM concentration, the Soa linkage on chromosome 6 (58 cM, lod score 9.4) was replicated, and another QTL was found on chromosome 19 (15 cM, lod score 3.2). Candidacy of previously identified genes in the close vicinity of the peak of the QTL was examined.


Prehemorrhage risk factors for fatal intracranial aneurysm rupture.
The goal of this study was to investigate predictive preictal risk factors for fatal subarachnoid hemorrhage (SAH) in a patient population with verified intracranial aneurysms without surgical selection of patients and with complete follow-up. A total of 142 patients with 181 unruptured aneurysms diagnosed between 1956 and 1978 were followed up for a total of 2577 person-years until death, SAH, or the years 1997 to 2000. The predictive value of several factors known before SAH was tested for case fatality. During follow-up, 34 first episodes of hemorrhage from a previously verified unruptured aneurysm occurred. Of these bleeding episodes, 17 were fatal. Patients who died after the bleeding had higher blood pressure values (mean+/-SD, 148+/-11/92+/-8 mm Hg; mean pressure, 111+/-9 mm Hg) before hemorrhage than did those with nonfatal bleeding (mean+/-SD, 135+/-15/83+/-11 mm Hg; mean, 101+/-12 mm Hg) (P<0.05). Patients with fatal SAH were also older (54+/-7 versus 47+/-13 years, P=0.068) and had aneurysms larger in diameter (13+/-8 versus 10+/-5 mm) than those who survived. They had a higher prevalence of definite hypertension (56% versus 12%, P<0.05), more frequently used antihypertensive medication (29% versus 6%) by the end of follow-up, and tended to have higher blood pressure at the beginning of follow-up (140+/-21/85+/-11 versus 134+/-17/80+/-9 mm Hg). After adjustment for age, aneurysm size, and sex, the only indisputable significant independent risk factor for fatal SAH compared with nonfatal SAH was systolic blood pressure before aneurysm rupture (odds ratio, 1.11 per 1 mm Hg; 95% CI, 1.01 to 1.23; P=0.032). The adjusted odds ratio of definite hypertension for fatal SAH was 12.67 (95% CI, 1.53 to 104.70; P=0.018). Increased systolic blood pressure values and long-term hypertension before aneurysm rupture seem to predict fatal SAH independently of aneurysm size or patient age or sex at the time of rupture.


The use of the Salt Stored Zona Assay in the study of the sperm function: methodological approach.
The authors mention many tests to study sperm function. On their opinion, the best model to evaluate spermo-oocyte interaction is the in vitro fertilization but it cannot be considered a test for ethical and practical problems. To overcome these problems they propose the Salt Stored Zona Assay to evaluate the spermatozoa.


Cultivation with untransfected fibroblasts stimulates proliferation of a single gene-modified fibroblast derived from a clawn miniature swine foetus.
Porcine embryonic fibroblasts (PEFs) have been used extensively as donor nuclei for the production of cloned pigs via somatic cell nuclear transfer (SCNT). Somatic cell nuclear transfer of gene-targeted PEFs has been the only way to produce gene-targeted pigs, given the lack of germ-line-competent porcine embryonic stem cells. Unlike other primary-cultured cells, such as murine embryonic fibroblasts, a single porcine PEF is unable to proliferate under normal conditions in which a certain number of PEFs (likely over 100) can grow normally. This limitation greatly hampers re-cloning of gene-modified PEFs, which is required for SCNT. Herein, we demonstrate the cultivation of a single PEF transfectant carrying the pEGFP-N1 plasmid with intact normal PEFs (>100) in a Terasaki microtest plate, which resulted in stimulation of the growth of the former cell (doubling time = 2.6 days). In contrast, when a single cell was cultured, it could typically divide only once and never divided more than twice. When a single transfectant was seeded in a well of a 96-well plate together with 5 × 10(4) untransfected PEFs and was subsequently selected in the presence of G418, we obtained a pure cell population of single-cell origin. Thus, this method should be useful for the purification of target recombinant pig clones from mosaic populations that cannot be cultured as a single cell or for which suitable cell growth-promoting conditions are unclear.


The potential role of spiral computed tomography in acute gastrointestinal bleeding.
We present four cases of profuse gastrointestinal (GI) bleeding where the site of haemorrhage was demonstrated using multi-slice computed tomography (MSCT), with concordant findings at laparotomy and pathological examination. A review of the current literature is undertaken and the role of spiral computed tomography as a method of sourcing the bleeding point is discussed in relation to other management modalities.


Tumor growth suppression by a retroviral vector displaying scFv antibody to CEA and carrying the iNOS gene.
In a recent in vitro study we demonstrated a specifically-targeted killing of CEA-expressing cells by a recombinant bifunctional retrovector displaying an scFv antibody to CEA and carrying the iNOS gene. In this study, we tested whether a gene therapy using the recombinant retrovirus could inhibit the growth of CEA-expressing tumors in mice. SCID mice were inoculated s.c. on the back with CEA-expressing MKN-45 cells on day 0. The recombinant viral particles were injected into the inoculated sites on days 3, 5 and 7 and tumor size was measured every 5 days. The s.c. administration of the recombinant retrovirus produced a marked growth inhibition of MKN-45 tumors in SCID mice. When the actual tumor weights were measured 50 days after initiation of treatment, about 70% reduction was observed in the treated group as compared to the control groups. This approach may also be applied to other tumor antigens expressed on cancer cells and is a step towards the cell specific suppression of tumorigenicity.


Purification of prosthetically intact sulfite oxidase from chicken liver using a modified procedure.
A modified procedure was used to purify sulfite oxidase (sulfite:O2 oxidoreductase; EC 1.8.3.1) from chicken liver in high yield. The modifications included dialysis of the enzyme against a buffered solution containing sodium molybdate (prior to ion-exchange chromatography), which apparently reconstituted any demolybdo enzyme present in the extract, and phenyl-Sepharose column chromatography. Analysis showed that the purified enzyme contained Mo and heme in a 1.03:1.00 ratio, indicating that the enzyme was prosthetically intact; exogenous heme and other colored proteins were absent from the final pool. Treatment of the sulfite-reduced enzyme with 50 mM cyanide at pH 8.5 resulted in a gradual loss of catalytic activity with a half-life of 19.7 min. Analysis of the cyanide-inactivated enzyme gave a Mo:heme ratio of 1.02:1.00, providing the first direct evidence that the enzyme does not lose molybdenum when inactivated with cyanide. This modified purification procedure provides enzyme in high yield which is well-suited for experiments requiring prosthetically intact enzyme and which is not contaminated with extraneous heme or with other redox active proteins.


Clinical and laboratory features of patients with rheumatoid arthritis diagnosed at rheumatology services in the Brazilian municipality of Cascavel, PR, Brazil.
Brazilian epidemiological studies on rheumatoid arthritis are scarce, thus all data currently available originate from the international literature. To determine the incidence and some clinical and laboratory characteristics of patients with rheumatoid arthritis in the municipality of Cascavel, state of Paraná, Brazil. Data were collected between August 2010 and July 2011 in all health services of the municipality of Cascavel that provided health care in Rheumatology: a university-affiliated hospital, a public outpatient clinic and four private clinics. We identified 38 patients diagnosed with rheumatoid arthritis, resulting in an estimated incidence of 13.4 cases per 100,000 inhabitants/year. Thirty two patients were females, whose mean age was 47.6 years. The age group with the highest incidence was over 40 years. The mean time between first symptoms and diagnosis was 12.4 months. Rheumatoid factor was positive in 68.4% of the patients, and 18.4% already had radiological abnormalities at diagnosis. The pharmacological treatment of patients was also assessed and proved to be in accordance with those found in the literature. The incidence of rheumatoid arthritis obtained in the municipality of Cascavel was lower than those reported in international studies.


Blastomycotic osteomyelitis of the pelvis: a case report.
Blastomycosis dermatitis is an unusual fungal infection that can become a diagnostic dilemma if a high index of suspicion is not maintained in patients who present with a chronic granulomatous infection of bone and soft tissue. We present a patient with a long history of pain, systemic manifestations of chronic infection, a draining sinus, and lytic changes on plain films to illustrate the difficulty in establishing the diagnosis. Appropriate staging studies and a bone biopsy led to the correct diagnosis in the patient.


G-CSF increases the number of peripheral blood dendritic cells CD16+ and modifies the expression of the costimulatory molecule CD86+.
Dendritic cells (DC) play a key role in initiating immune reactions after allogeneic stem cell transplantation. The two main peripheral blood DC populations are myeloid (DC1) and lymphoplasmacytoid (DC2). A new subset of myeloid DC, expressing CD16, has been identified. We analyzed the number and CD86 expression of DC subsets in peripheral blood of 18 healthy donors, before and after granulocyte colony-stimulating factor (G-CSF) and in the inoculum of allogeneic peripheral blood transplants (allo-PBT; n=100) and allogeneic bone marrow transplants (allo-BMT; n=22). Granulocyte colony-stimulating factor administration increased the median number of DC1 (P=0.0007), of DC2 (P<0.0001) and of DC CD16+ (P=0.0001). Granulocyte colony-stimulating factor administration was also associated with a significant decrease of CD86 expression on DC1 (P=0.0003) and with a trend for an increase on DC CD16+ (P=0.07). Recipients of allo-PBT received similar quantities of DC1 and higher doses of DC2 and DC CD16+ than recipients of allo-BMT (P=0.5; P=0.0001; P<0.0001, respectively). Granulocyte colony-stimulating factor modifies the number of DC in peripheral blood and the expression of the costimulatory molecule CD86. This resulted in a different composition of DC2 and especially of DC CD16+ in the harvests, which might explain some of the differences observed in allogeneic reactions after allo-PBT with respect to allo-BMT.


The lipid peroxidation product 4-hydroxynonenal is formed by--and is able to attract--rat neutrophils in vivo.
4-Hydroxynonenal (HNE), a major aldehydic product of lipid peroxidation, is a chemoattractant for neutrophilic polymorphonuclear granulocytes in vitro. The question was studied, whether HNE is formed during the ingress of neutrophils in the Sephadex model of inflammation. The polydextrane Sephadex G-200, which causes an acute aseptic traumatic inflammation, was injected subcutaneously into rats. The implants were excised 6-36 hours later, and the neutrophils separated from the exsudate by centrifugation. After extraction with dichloromethane HNE was identified in the exsudate by non-derivative reversed phase HPLC in combination with on-line uv-spectroscopy. The concentration of HNE in the inflammatory focus did not correlate with the number of neutrophils present. While the peak of HNE coincided with the time point of the highest turnover rate of neutrophils (0.13 microM at 6 hrs after implantation), the highest number of neutrophils (about 100 million cells) occurred not earlier than 18 hrs later (24 hrs after onset of inflammation). When neutrophils were isolated from the inflammatory focus and stimulated with Zymosan, they were able to produce HNE in vitro depending on the time of isolation. The highest production of HNE (0.17 microM) by phagocyting neutrophils was observed at the shortest inflammation time studied (3 hrs). In order to compare these results with the oxidative burst of neutrophils the formation of superoxide was also measured by the cytochrome c reduction assay in vitro. The maximum of the production rate of superoxide anion was observed at the same inflammation time (6 hrs), when the HNE maximum occurred.(ABSTRACT TRUNCATED AT 250 WORDS)


Alteration of thiol-disulfide homeostasis in workers occupationally exposed to arsenic.
This study evaluates the redox state in occupationally arsenic-exposed workers (n = 71) by assessing the dynamic serum thiol-disulfide homeostasis. We determined the serum thiol-disulfide homeostasis parameters of exposed workers and controls (n = 43) using a novel automated colorimetric assay. Median urinary As and 8-isoprostane levels of exposed group were significantly higher than control group (16.40 μg/L vs 2μg/L, p < .001 and 2.28 ng/ml vs 0.54 ng/ml, p < .001, respectively). Disulfide level, disulfide/native thiol ratio, and disulfide/total thiol ratio were significantly higher in exposed group. The mean ceruloplasmine and myeloperoxidase activities of As-exposed group were significantly higher than control group (117.15 U/L vs 87.02 U/L, p = .035 and 148.53 U/L vs 97.75 U/L, p = .000, respectively). The median catalase activity did not differ in the two groups. Our findings revealed that As disrupts the thiol-disulfide homeostasis in favor of disulfide.


Diffusion-weighted magnetic resonance imaging for detecting and assessing ileal inflammation in Crohn's disease.
Whether diffusion-weighted imaging (DWI)-MRI is of value in detecting and assessing inflammation of ileal Crohn's disease (CD) remains poorly investigated. To compare DWI-MR enterography (MRE) with conventional MRE in estimating inflammation in small bowel CD, to determine an apparent diffusion coefficient (ADC) threshold to differentiate active from non-active lesions and to assess inter-observer agreement. Thirty-one CD patients from the Clermont-Ferrand IBD unit with ileal involvement were consecutively and prospectively included between April and June 2011. All patients underwent DWI-MRI to detect the digestive segment with the most severe lesions, which was then used to calculate the ADC. Qualitative and quantitative results were compared with conventional MRE including MaRIA (Magnetic Resonance Index of Activity) score calculation and independent activity predictors (wall thickening, oedema, ulcers). Each examination was interpreted independently by two radiologists blinded for clinical assessment. Seventeen patients (54.8%) had active CD as defined by the MaRIA score ≥7. DWI hyperintensity was highly correlated with disease activity evaluated using conventional MRE (P = 0.001). Qualitative analysis of DW sequences determined sensitivity, specificity, positive predictive value and negative predictive value as 100%, 92.9%, 94.4% and 100% respectively. Quantitative analysis using a cut-off of 1.6 × 10(-3) mm(2)/s for ADC yielded sensitivity and specificity values of, respectively, 82.4% and 100%. Inter-observer agreement was high with regard to DWI hyperintensity (κ = 0.69, accuracy rate = 85.7%) and ADC (correlation = 0.74, P < 0.001, and concordance = 0.71, P < 0.001). DWI-MR enterography is a well-tolerated, non-time-consuming and accurate tool for detecting and assessing inflammation in small bowel Crohn's disease.


Intrasulcular laser Doppler readings before and after root planing.
A reproducible and sensitive laser Doppler periodontal probe has recently been developed for intrasulcular measurement of gingival blood flow. The specific aims of this investigation were to determine the relation between intrasulcular laser Doppler readings (LDR) and traditional diagnostic criteria as well as to evaluate the response to root planning in terms of LDR and traditional criteria. LDR and clinical measurements (bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL) were obtained from 2 healthy and 2 diseased sites in 30 systemically healthy adult volunteers with localized moderate to advanced periodontitis. All 30 subjects were re-examined 1 month following root planing while 10 subjects were re-examined at approximately 1 year after treatment. Subject-adjusted correlations between pretreatment LDR and PD as well as LDR and CAL were 0.74 and 0.71, respectively. 1 month following root planing, the diseased sites had undergone a significant reduction in LDR and PD with an accompanying gain in CAL. Prior to treatment, 95 of 120 sites (79%) agreed on an ordinal classification (high, low) for LDR and BOP. Mantel-Haenszel common odds ratios for agreement between LDR and BOP were 9.6 pre-treatment and 4.3 one month after treatment. A slight rebound of all measurements was noted in a group of 10 subjects followed for 1 year. It was concluded that the laser Doppler periodontal probe is an unbiased non-invasive method of monitoring the response to periodontal therapy.


Extreme mtDNA divergences in a terrestrial slug (Gastropoda, Pulmonata, Arionidae): accelerated evolution, allopatric divergence and secondary contact.
Extremely high levels of intraspecific mtDNA differences in pulmonate gastropods have been reported repeatedly and several hypotheses to explain them have been postulated. We studied the phylogeny and phylogeography of 51 populations (n = 843) of the highly polymorphic terrestrial slug Arion subfuscus (Draparnaud, 1805) across its native distribution range in Western Europe. By combining the analysis of single stranded conformation polymorphisms (SSCP) and nucleotide sequencing, we obtained individual sequence data for a fragment of the mitochondrial 16S rDNA and a fragment of the nuclear ITS1. Additionally, five polymorphic allozyme loci were scored. Based on the 16S rDNA phylogeny, five monophyletic haplotype groups with sequence divergences of 9-21% were found. Despite this deep mitochondrial divergence, the haplotype groups were not monophyletic for the nuclear ITS1 fragment and haplotype group-specific allozyme alleles were not found. Although there is evidence for an accelerated mtDNA clock, the divergence among the haplotype groups is older than the Pleistocene and their current allopatric ranges probably reflect allopatric divergence and glacial survival in separate refugia from which different post-glacial colonization routes were established. A range-overlap of two mtDNA groups (S1 and S2, 21% sequence divergence) stretched from Central France and Belgium up to the North of the British Isles. The nuclear data suggest that this secondary contact resulted in hybridization between the allopatrically diverged groups. Therefore, it seems that, at least for two of the groups, the deep mtDNA divergence was only partially accompanied by the formation of reproductive isolation.


Surgical approach to tumor recurrence and metastatic lesions of the liver in a patient with malignant endocrine tumors of the pancreas: case report.
We describe herein a 72-year-old woman with tumor recurrence in the residual pancreas and metastasis to the liver following a pylorus-preserving pancreatoduodenectomy for multiple endocrine tumors in the head of the pancreas. Abdominal ultrasonography performed 7 years after the initial surgery detected new lesions in the residual pancreas and liver. After recurrence of endocrine tumors of the pancreas and metastasis to the liver were diagnosed, the lesions were successfully resected by total pancreatectomy with distal gastrectomy and both lateral segmentectomy and partial resection of segment 8. Genetic analysis using a blood specimen showed that this patient carried the multiple endocrine neoplasia type 1 (MEN1) gene mutation. One year after the second resection, the patient remains in good health using insulin and has not shown any sign of recurrence. This case report describes successful surgical resection for recurrence and metastasis of malignant endocrine tumors in a patient with the MEN1 gene mutation.


Association Between Dietary Selenium Intake and the Prevalence of Nonalcoholic Fatty Liver Disease: A Cross-Sectional Study.
Objective: The aim was to examine the association between dietary selenium intake and nonalcoholic fatty liver disease (NAFLD) in a large group of middle-aged and elderly Chinese persons. Method: The data included in this analysis were from a population-based study, the Xiangya Hospital Health Management Center Study. NAFLD was diagnosed by (1) imaging or histological evidence of hepatic steatosis; (2) absence of specific etiologies of NAFLD; and (3) no heavy consumption of alcohol. Dietary selenium intake was assessed using a validated semi-quantitative food frequency questionnaire. The association between dietary selenium intake and the prevalence of NAFLD was evaluated using logistic and spline regression in a cross-sectional study of 5436 subjects. Results: The prevalence of NAFLD was 36.8%. Compared with the lowest quintile, the energy-adjusted odds ratios for NAFLD were 1.27 (95% confidence interval [CI], 1.07-1.52), 1.30 (95% CI, 1.09-1.55), and 1.58 (95% CI, 1.33-1.89) for the third, fourth, and fifth quintiles of selenium intake, respectively, and there was a positive dose-response relationship (r = 0.88, p for trend = 0.008). Similar results were observed for men and women separately. The findings were not materially altered by adjustment for potential confounders (i.e., age, gender, body mass index, smoking status, diabetes, hypertension, activity level, nutritional supplements, energy intake, fat intake, fiber intake, cholesterol and saturated fatty acid intake). Conclusions: In this middle-aged and elderly population, subjects with higher dietary selenium intake, even below the recommended nutrient intake in China, had higher prevalence of NAFLD in a dose-response relationship manner.


Cerebrovascular responses under controlled and monitored physiological conditions in the anesthetized mouse.
Control of physiological parameters such as respiration, blood pressure, and arterial blood gases has been difficult in the mouse due to the lack of technology required to monitor these parameters in small animals. Here we report that anesthetized and artificially ventilated mice can be maintained under physiological control for several hours with apparently normal cerebrovascular reactivity to hypercapnia and mechanical vibrissal stimulation. SV-129 mice were anesthetized with urethane (750 mg/kg i.p.) and alpha-chloralose (50 mg/kg i.p.), intubated, paralyzed, and artificially ventilated. Respiratory control was maintained within physiological range by reducing the inspiratory phase of the respiratory cycle to < 0.1 s and by adjusting end-tidal CO2 to give a PCO2 of 35 +/- 3 mm Hg. In these mice, mean arterial pressure (95 +/- 9 mm Hg), heart rate (545 +/- 78 beats/min), and arterial pH (7.27 +/- 0.10) could be maintained for several hours. Body temperature was kept at 36.5-37.5 degrees C. We observed stable regional CBF (rCBF) measurements (as determined by laser-Doppler flowmetry) when systemic arterial blood pressure was varied between 40 and 130 mm Hg. Hypercapnia led to a 38 +/- 15% (5% CO2) and 77 +/- 34% (10% CO2) increase in rCBF. Mechanical stimulation of contralateral vibrissae for 1 min increased rCBF by 14 +/- 4%. Changes in rCBF compare favorably with those observed previously in another rodent species, the Sprague-Dawley rat. After placement of a closed cranial window, cerebrovascular reactivity to hypercapnia and whisker stimulation was intact and well maintained during 2-h superfusion with artificial CSF.


Electrochemistry at One Nanoparticle.
Electrochemistry at metal nanoparticles (NPs) is of significant current interest because of its applications in catalysis, energy conversion and storage, and sensors. The electrocatalytic activity of NPs depends strongly on their size, shape, and surface attachment. The use of a large number of particles in most reported kinetic experiments obscured the effects of these factors because of polydispersity and different NP orientations. Recent efforts to probe electrochemistry at single NPs included recording of the catalytically amplified current produced by random collisions of particles with the electrode surface, immobilizing an NP on the surface of a small electrode, and delivering individual NPs to electrode surfaces. Although the signals recorded in such experiments were produced by single NPs, the characterization issues and problems with separating an individual particle from other NPs present in the system made it difficult to obtain spatially and/or temporally resolved information about heterogeneous processes occurring at a specific NP. To carry out electrochemical experiments involving only one NP and characterize such an NP in situ, one needs nanoelectrochemical tools with the characteristic dimension smaller than or comparable to those of the particle of interest. This Account presents fundamentals of two complementary approaches to studying NP electrochemistry, i.e., probing single immobilized NPs with the tip of a scanning electrochemical microscope (SECM) and monitoring the collisions between one catalytic NP and a carbon nanopipette. The former technique can provide spatially resolved information about NP geometry and measure its electron transfer properties and catalytic activity under steady-state conditions. The emphasis here is on the extraction of quantitative physicochemical information from nanoelectrochemical data. By employing a polished disk-type nanoelectrode as an SECM tip, one can characterize a specific nanoparticle in situ and then use the same NP for kinetic experiments. A new mode of SECM operation based on tunneling between the tip and nanoparticle can be used to image the NP topography with a lateral resolution of ∼1 nm. An alternative approach employs carbon nanoprobes produced by chemical vapor deposition of carbon into quartz nanopipettes. One metal NP is captured inside the carbon nanocavity to probe the dynamics of its interactions with the electrode surface on the microsecond time scale. The use of high-resolution transmission electron microscopy is essential for interpreting the results of single-NP collision experiments. A brief discussion of the nanoelectrochemical methodology, recent advances, and future directions is included.


[The effect of cerebrocrast on the microcirculation in acute transient cerebral ischemia].
In experiments on rats with cerebral ischemia (30-min occlusion of carotid arteries), the novel calcium antagonist Cerebrocrast (0.4 microgram.kg-1.min-1 i.v. for 15 min) was found to provide the functional maintenance of the parietal cerebral bed in the postischemic period by improving cerebral hemodynamics, reducing the number of highly narrowed capillaries. The agent prevented the erythrocytic accumulation of lipid peroxidation (LPO) products (diene conjugates and Schiff bases) in the intra- and postischemic periods, promoted the maintenance of viscoelastic properties during LPO activation.


Refractive errors and binocular dysfunctions in a population of university students.
This clinical study was performed to determine the presence of refractive errors and binocular dysfunctions in a population of university students. Refraction and binocular function were evaluated in a young patient population (230 students and 234 nonstudent subjects, aged 18-27 years). Distance visual acuity (DVA) and near visual acuity (NVA), refraction, cover test (CT), ocular motility, near-point of convergence, horizontal phoria measurement by Maddox wing, negative and positive vergence amplitude in prism diopters, fusion amplitude in synoptophore, as well as stereoacuity (Titmus test) were tested. Emmetropia was the most frequent refractive status in our student and nonstudent groups (78.7%). Myopia was the most frequent refractive disorder in the whole population (13.1%). Myopia and hypermetropia were significantly more frequent in the students than in nonstudents (chi-square emp 47.55). Exophoria is significantly more frequent in myopic subjects. Vergence amplitude (t test 0.000) and fusion amplitude (t test 0.005) show significantly lower values in student population. Results of Titmus test in the student group is significantly worse than in the nonstudent group (t test 0.000). Maddox wing resulted in significantly higher degree of heterophoria in the student population (t test 0.000). Myopic subjects, in the student group (t test 0.002) as well as in the nonstudent group (t test 0.001), show significantly better results in Titmus test. High near visual demand could be the most important factor for higher incidence of myopia, worse convergence and fusion amplitude, higher degree of exophoria, and worse results in Titmus test in the student population.


Increasing the dimensionality of cryogenic molecular coolers: Gd-based polymers and metal-organic frameworks.
The magnetothermal properties of a coordination polymer and a metal-organic framework (MOF) based on Gd(3+) ions are reported. An equally large cryogenic magnetocaloric effect (MCE) is found, irrespective of the dimensionality. This combined with their robustness makes them appealing for widespread magnetic refrigeration applications.


Heterogeneity of Macrophages in Atherosclerosis.
Atherosclerosis is a prevalent inflammatory condition and a frequent cause of morbidity and mortality worldwide. Macrophages are among the key immune cells driving lesion formation in the arterial wall. They have therefore evolved as potential targets for therapeutic strategies. Understanding of the different macrophage phenotypes and functions seems to be of pivotal importance for the development of treatments to target these immune cells. This review highlights the complexity of the mononuclear phagocyte system and summarizes important features of macrophage biology contributing to atherosclerosis.


Use of non-physiological buffer systems in the analysis of methotrexate transport in L1210 cells.
Methotrexate transport parameters have been compared in L1210 cells suspended in a series of HEPES buffer systems of varying ionic compositions. While no effect was observed on the Vmax for methotrexate influx, the Kt for half-maximal influx, the steady-state level of methotrexate, and the efflux rate each varied substantially and to an extent which could be correlated directly to the anionic composition of the external medium. Buffer composition also affected the membrane potential, the ATP level of the cells, and, in one instance, the cell volume, but these changes did not exert a significant effect on the transport process. These results suggest that the integrity of L1210 cells is not adversely affected by either the presence of HEPES in the suspending medium or by the absence of certain physiological ions, and, moreover, that methotrexate transport parameters measured under these conditions, although not necessarily indicative of the quantitative events that might occur in vivo, can nevertheless provide meaningful information on the properties and mechanism of this transport system.


Identification of coenzyme M biosynthetic phosphosulfolactate synthase: a new family of sulfonate-biosynthesizing enzymes.
The hyperthermophilic euryarchaeon Methanococcus jannaschii uses coenzyme M (2-mercaptoethanesulfonic acid) as the terminal methyl carrier in methanogenesis. We describe an enzyme from that organism, (2R)-phospho-3-sulfolactate synthase (ComA), that catalyzes the first step in coenzyme M biosynthesis. ComA catalyzed the stereospecific Michael addition of sulfite to phosphoenolpyruvate over a broad range of temperature and pH conditions. Substrate and product analogs moderately inhibited activity. This enzyme has no significant sequence similarity to previously characterized enzymes; however, its Mg(2+)-dependent enzyme reaction mechanism may be analogous to one proposed for enolase. A diverse group of microbes and plants have homologs of ComA that could have been recruited for sulfolactate or sulfolipid biosyntheses.


Modulation of deoxynucleotide metabolism by the deoxycytidylate deaminase inhibitor 3,4,5,6-tetrahydrodeoxyuridine.
Tetrahydrodeoxyuridine (dTHU) inhibits deoxycytidine deaminase and, after intracellular phosphorylation to the active 5'-monophosphate, also inhibits deoxycytidylate deaminase (dCMPD). Because in vitro studies have shown that dCMPD may regulate pyrimidine deoxynucleotide metabolism, the objective of this study was to investigate the effects of dTHU on deoxynucleotide metabolism in whole cells. Nearly complete inhibition of dCMPD, measured in intact CCRF-CEM cells by incorporation of [14C]dCyd into dTTP, occurred after a 45-min incubation with 100 microM dTHU. This was accompanied by an 8-fold dCTP pool expansion, although dATP, dTTP, dGTP, and ribonucleoside triphosphate pools were unaffected. Tetrahydrouridine, which inhibits deoxycytidine deaminase exclusively, had no effect on nucleotide pools. The dCTP pool expansion was directly proportional to the dTHU concentration (3-100 microM) and reached a maximum after 2 hr. Inhibition of ribonucleotide reductase by hydroxyurea completely prevented the dTHU-induced dCTP pool expansion, indicating that the substrate of dCMPD was derived from the ribonucleotide pool and that CDP was the predominant precursor of dCTP. dTHU-mediated inhibition of dCMPD appeared reversible. Exposure of cells to 100 microM dTHU followed by washing into fresh medium resulted in a linear decrease of the dCTP pool and an increase in the dTTP pool. The increased dCTP concentration after preincubation with dTHU was associated with an inhibition of deoxycytidine kinase, as indicated by a reduced capacity of cells to phosphorylate ara-C. dTHU is a useful new tool for investigating the role of dCMPD in the regulation of deoxynucleotide metabolism in whole cells.


Effect of Deformity and Malunion of the Anterior Pelvic Ring.
Pelvic fractures are common after high-energy trauma and are often associated with ligamentous injury. Treatment is guided by assessing stability of the pelvic ring, and unstable injuries frequently require surgery to achieve a desirable outcome. Assessment of pelvic ring stability is often possible with physical examination and standard imaging studies (plain radiographs and computed tomography); however, these "static" imaging modalities may not adequately identify dynamically unstable pelvic injuries that require surgery. Cadaveric and clinical data suggest that the injured pelvis may recoil significantly from the point of maximal displacement, and some unstable injuries may not be recognized until patients present with clinical symptoms. This article presents the case of a patient who sustained a minimally displaced pelvic ring injury that was stable on bedside examination and static imaging, but ultimately was unstable. She developed a substantial pelvic malunion with significant pain and activity limitations. The patient subsequently underwent successful pelvic ring reconstruction, and she remains asymptomatic at 2 years.


Creeping attachment: autogenous graft vs dermal matrix allograft.
For many years, free autogenous grafts have been used as a method of gaining keratinized tissue around teeth with mucogingival problems. Creeping attachment using autogenous graft material has been actively studied. In addition, biocompatible, acellular connective-tissue material has recently been used as an alternative to free gingival grafts to increase the zone of keratinization. This report presents a patient with bilateral mucogingival defects in the canine and premolar areas. The patient received an autogenous graft on one side and a dermal matrix allograft on the contralateral side. Creeping attachments were measured and compared at 3 months and 12 months after surgery. After 12 months of healing, an average of 1.23 mm of creeping attachment was measured on the free gingival graft side and 0.96 mm of creeping attachment was measured with the dermal matrix allograft.


Not your average bigot: The better-than-average effect and defensive responding to Implicit Association Test feedback.
A robust body of literature on the better-than-average effect suggests that people believe that they are better than the average others across a variety of domains. In two studies, we examined whether these better-than-average beliefs occur for bias related to stereotyping and prejudice. Moreover, we investigated the hypothesis that better-than-average beliefs will predict defensive responding to feedback indicating personal bias (e.g., preferences for majority groups, societally endorsed stereotypes). Specifically, we examined defensive responses to implicit attitude feedback. Study 1 examined this prediction using archival analysis of two large, online samples of participants completing a Weight-related Implicit Association Test (IAT). Study 2 conceptually replicated Study 1 using nine different, randomly assigned IATs and additional measures of defensiveness. In both studies, people generally believed that they were less biased than others. Moreover, people responded defensively to feedback indicating they were biased. This effect was moderated by better-than-average beliefs such that feedback indicating societally consistent bias was related to defensiveness most (and sometimes only) when people believed they were better than average initially. This work represents the first foray into examining the possible moderating role of social-comparative beliefs in predicting responses to implicit attitude feedback and spurs several important avenues for future research.


Phylogeography of the common goby, Pomatoschistus microps, with particular emphasis on the colonization of the Mediterranean and the North Sea.
The phylogeographical patterns of a small marine fish, the common goby, Pomatoschistus microps, were assessed at 12 sites along the northeastern Atlantic coasts and the western Mediterranean Sea. A combination of two genetic markers was employed: cellulose acetate allozyme electrophoresis (CAGE) and sequence analysis of a 289 bp fragment of the mitochondrial locus cytochrome b. Both markers were congruent in revealing significant differences between samples (global FST = 0.247 for the allozymes and PhiST = 0.437 for the mitochondrial DNA data) and a pattern of isolation-by-distance. Phylogeographical analyses yielded a shallow branching structure with four groups. Three of those were confined to the Atlantic basin and showed a star-like pattern. The fourth group contained a central haplotype occurring at the edges of the species' distribution, accompanied by a few more rare variants, which were restricted to the Mediterranean Sea. A genetic break was observed around the British Isles, with distinct haplotypes dominating at either side of the English Channel. A significantly negative correlation between the degree of genetic diversity and latitude was recorded both for mitochondrial DNA (mtDNA) and allozymes in the Atlantic basin. Gene flow analysis suggested that recolonization of the North Sea and the coasts of western Scotland and Ireland may have taken place from a glacial refugium in the Southern Bight of the North Sea. These results are discussed in the perspective of possible postglacial migration routes of marine fish along the northeastern Atlantic coasts.


Aging and the pharmacokinetics and metabolism of metoprolol enantiomers in the rat.
After i.v. administration of racemic metoprolol in the rat, the plasma concentrations of (R)- and (S)-metoprolol were comparable, and no differences in pharmacokinetic parameters between the two enantiomers were found. From the 3rd to the 12th month, comparable changes were seen for both enantiomers: there was an increase in the area under the plasma concentration-time curve (AUC) and a decrease in blood and plasma clearance. Half-life showed a significant prolongation, volume of distribution decreased between 3 and 12 months and increased between 12 and 24 months. After oral administration of the racemate, AUC and Cmax (maximum plasma concentration) were slightly higher, while oral clearance was slightly lower for (R)-metoprolol than for (S)-metoprolol. With aging, Cmax and AUC increased for both enantiomers, while oral clearance decreased. The change in oral clearance as a function of age is different between (S)- and (R)-metoprolol, and thus enantioselective. In vitro disappearance rate in 3-month-old rats was significantly higher for (S)-metoprolol than for (R)-metoprolol, although the difference was small. With aging, the disappearance rates of both enantiomers increased significantly, but not enantioselectively.


The regulation of host cellular and gut microbial metabolism in the development and prevention of colorectal cancer.
Metabolism regulation is crucial in colorectal cancer (CRC) and has emerged as a remarkable field currently. The cellular metabolism of glucose, amino acids and lipids in CRC are all reprogrammed. Each of them changes tumour microenvironment, modulates bacterial composition and activity, and eventually promotes CRC development. Metabolites such as short chain fatty acids, secondary bile acids, N-nitroso compounds, hydrogen sulphide, polyphenols and toxins like fragilysin, FadA, cytolethal distending toxin and colibactin play a dual role in CRC. The relationship of gut microbe-metabolite is essential in remodelling intestinal microbial ecology composition and metabolic activity. It regulates the metabolism of colonic epithelial cells and changes the tumour microenvironment in CRC. Microbial metabolism manipulation has been considered to be potentially preventive in CRC, but more large-scale clinical trials are required before their application in clinical practice in the near future.


Insights into the olfactory system of the ectoparasite Caligus rogercresseyi: molecular characterization and gene transcription analysis of novel ionotropic receptors.
Although various elements of the olfactory system have been elucidated in insects, it remains practically unstudied in crustaceans at a molecular level. Among crustaceans, some species are classified as ectoparasites that impact the finfish aquaculture industry. Thus, there is an urgent need to identify and comprehend the signaling pathways used by these in host recognition. The present study, through RNA-seq and qPCR analyses, found novel transcripts involved in the olfactory system of Caligus rogercresseyi, in addition to the transcriptomic patterns expressed during different stages of salmon lice development. From a transcriptomic library generated by Illumina sequencing, contigs that annotated for ionotropic receptors and other genes implicated in the olfactory system were identified and extracted. Full length mRNA was obtained for the ionotropic glutamate receptor 25, which had 3923 bp, and for the glutamate receptor ionotropic kainate 2, which had 2737 bp. Furthermore, two other transcripts identified as glutamate receptor, ionotropic kainate 2-like were found. In silico analysis was performed for the transcription expression from different stages of development in C. rogercresseyi, and clusters according to RPKM values were constructed. Gene transcription data were validated through qPCR assays in ionotropic receptors, and showed an expression of glutamate receptor 25 associated with the copepodid stage whereas adults, especially male adults, were associated with the kainate 2 and kainate 2-like transcripts. Additionally, gene transcription analysis of the ionotropic receptors showed an overexpression in response to the presence of masking compounds and immunostimulant in salmon diets. This response correlated to a reduction in sea lice infection following in vivo challenge. Diets with masking compounds showed a decrease of lice infestation of up to 25%. This work contributes to the available knowledge on chemosensory systems in this ectoparasite, providing novel elements towards understanding the host-finding process of the salmon louse C. rogercresseyi.