Serum microRNA signatures as "liquid biopsies" for interrogating hepatotoxic mechanisms and liver pathogenesis in human.
MicroRNAs (miRNAs) released into the peripheral circulation upon cellular injury have shown a promise as a new class of tissue-specific biomarkers. We were first to demonstrate that next-generation sequencing analysis of serum from human subjects with acetaminophen-induced liver injury revealed a specific signature of circulating miRNAs. We consequently hypothesized that different types of hepatic liver impairments might feature distinct signatures of circulating miRNAs and that this approach might be useful as minimally invasive diagnostic "liquid biopsies" enabling the interrogation of underlying molecular mechanisms of injury in distant tissues. Therefore we examined serum circulating miRNAs in a total of 72 serum samples from a group of 53 subjects that included patients with accidental acetaminophen overdose, hepatitis B infection, liver cirrhosis and type 2 diabetes as well as gender- and age-matched healthy subjects with no evidence of liver disease. The miRNA signatures were identified using next-generation sequencing that provided analysis for the whole miRNome, including miRNA isoforms. Compared to the healthy subjects, a total of 179 miRNAs showed altered serum levels across the diseased subjects. Although many subjects have elevated alanine aminotransferase suggesting liver impairments, we identified distinct miRNA signatures for different impairments with minimum overlap. Furthermore, the bioinformatics analysis of miRNA signatures revealed relevant molecular pathways associated with the mechanisms of toxicity and or pathogenesis of disease. Interestingly, the high proportion of miRNA isoforms present in the respective signatures indicated a new level of complexity in cellular response to stress or disease. Our study demonstrates for the first time that signatures of circulating miRNAs show specificity for liver injury phenotypes and, once validated, might become useful for diagnosis of organ pathologies as "liquid biopsies".


DNA aptamers against bacterial cells can be efficiently selected by a SELEX process using state-of-the art qPCR and ultra-deep sequencing.
DNA aptamers generated by cell-SELEX against bacterial cells have gained increased interest as novel and cost-effective affinity reagents for cell labelling, imaging and biosensing. Here we describe the selection and identification of DNA aptamers for bacterial cells using a combined approach based on cell-SELEX, state-of-the-art applications of quantitative real-time PCR (qPCR), next-generation sequencing (NGS) and bioinformatic data analysis. This approach is demonstrated on Enterococcus faecalis (E. faecalis), which served as target in eleven rounds of cell-SELEX with multiple subtractive counter-selections against non-target species. During the selection, we applied qPCR-based analyses to evaluate the ssDNA pool size and remelting curve analysis of qPCR amplicons to monitor changes in pool diversity and sequence enrichment. Based on NGS-derived data, we identified 16 aptamer candidates. Among these, aptamer EF508 exhibited high binding affinity to E. faecalis cells (KD-value: 37 nM) and successfully discriminated E. faecalis from 20 different Enterococcus and non-Enterococcus spp. Our results demonstrate that this combined approach enabled the rapid and efficient identification of an aptamer with both high affinity and high specificity. Furthermore, the applied monitoring and assessment techniques provide insight into the selection process and can be highly useful to study and improve experimental cell-SELEX designs to increase selection efficiency.


Manufacture of Platform Chemicals from Pine Wood Polysaccharides in Media Containing Acidic Ionic Liquids.
Pinus pinaster wood samples were subjected to chemical processing for manufacturing furans and organic acids from the polysaccharide fractions (cellulose and hemicellulose). The operation was performed in a single reaction stage at 180 or 190 °C, using a microwave reactor. The reaction media contained wood, water, methyl isobutyl ketone, and an acidic ionic liquid, which acted as a catalyst. In media catalyzed with 1-butyl-3-methylimidazolium hydrogen sulfate, up to 60.5% pentosan conversion into furfural was achieved, but the conversions of cellulose and (galacto) glucomannan in levulinic acid were low. Improved results were achieved when AILs bearing a sulfonated alkyl chain were employed as catalysts. In media containing 1-(3-sulfopropyl)-3-methylimidazolium hydrogen sulfate as a catalyst, near quantitative conversion of pentosans into furfural was achieved at a short reaction time (7.5 min), together with 32.8% conversion of hexosans into levulinic acid. Longer reaction times improved the production of organic acids, but resulted in some furfural consumption. A similar reaction pattern was observed in experiments using 1-(3-sulfobutyl)-3-methylimidazolium hydrogen sulfate as a catalyst.


Integration and expression of viral DNA in cells transformed by host range mutants of adenovirus type 5.
Group I host range (hr) mutants of adenovirus type 5 are unable to transform rat embryo or rat embryo brain cells but induce an abnormal transformation of baby rat kidney cells. We established several transformed rat kidney cell lines and characterized them with respect to the transformed phenotype and the structure of the integrated viral DNA. The hr mutant-transformed cells, unlike wild-type virus transformants, were fibroblastic rather than epithelial, failed to grow in soft agar, and were also less tumorigenic in nude mice. Studies on the structure of the integrated viral DNA sequences showed that hr-transformed cells always contained the left end of the adenovirus DNA, but the size of the integrated DNA fragment varied among different lines, and a high percentage of the lines contained the entire viral genome colinearly integrated. The patterns of integration were maintained after prolonged growth in culture and after subcloning. Attempts to rescue infectious virus from lines which contained the entire genome were unsuccessful. Using immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, we analyzed the viral proteins expressed in hr-transformed cells. Results of these studies indicated that, like wild type-transformed cells, hr transformants expressed E1B proteins of molecular weight 58,000 and 19,000.


High-performance hardware implementation of a parallel database search engine for real-time peptide mass fingerprinting.
Peptide mass fingerprinting (PMF) is a method for protein identification in which a protein is fragmented by a defined cleavage protocol (usually proteolysis with trypsin), and the masses of these products constitute a 'fingerprint' that can be searched against theoretical fingerprints of all known proteins. In the first stage of PMF, the raw mass spectrometric data are processed to generate a peptide mass list. In the second stage this protein fingerprint is used to search a database of known proteins for the best protein match. Although current software solutions can typically deliver a match in a relatively short time, a system that can find a match in real time could change the way in which PMF is deployed and presented. In a paper published earlier we presented a hardware design of a raw mass spectra processor that, when implemented in Field Programmable Gate Array (FPGA) hardware, achieves almost 170-fold speed gain relative to a conventional software implementation running on a dual processor server. In this article we present a complementary hardware realization of a parallel database search engine that, when running on a Xilinx Virtex 2 FPGA at 100 MHz, delivers 1800-fold speed-up compared with an equivalent C software routine, running on a 3.06 GHz Xeon workstation. The inherent scalability of the design means that processing speed can be multiplied by deploying the design on multiple FPGAs. The database search processor and the mass spectra processor, running on a reconfigurable computing platform, provide a complete real-time PMF protein identification solution.


How Do We Value Postoperative Recovery?: A Systematic Review of the Measurement Properties of Patient-reported Outcomes After Abdominal Surgery.
To appraise the level of evidence supporting the measurement properties of patient-reported outcome measures (PROMs) in the context of postoperative recovery after abdominal surgery. There is growing interest in using PROMs to support value-based care in abdominal surgery; however, to draw valid conclusions regarding patient-reported outcomes data, PROMs with robust measurement properties are required. Eight databases (MEDLINE, EMBASE, Biosis, PsycINFO, The Cochrane Library, CINAHL, Scopus, Web of Science) were searched for studies focused on the measurement properties of PROMs in the context of recovery after abdominal surgery. The methodological quality of individual studies was evaluated using the consensus-based COSMIN checklist. Evidence supporting the measurement properties of each PROM was synthetized according to standardized criteria and compared against the International Society of Quality of Life Research minimum standards for the selection of PROMs for outcomes research. We identified 35 studies evaluating 22 PROMs [12 focused on nonspecific surgical populations (55%), 4 focused on abdominal surgery (18%), and 6 generic PROMs (27%)]. The great majority of the studies (74%) received only poor or fair quality ratings. Measurement properties of PROMs were predominantly supported by limited or unknown evidence. None of the PROMs fulfilled International Society of Quality of Life Research's minimum standards, hindering specific recommendations. There is very limited evidence supporting the measurement properties of existing PROMs used in the context of recovery after abdominal surgery. This precludes the use of these PROMs to support value-based surgical care. Further research is required to bridge this major knowledge gap. International Prospective Register of Systematic Reviews (PROSPERO): CRD42014014349.


Characterization of Tissue Transglutaminase as a Potential Biomarker for Tissue Response toward Biomaterials.
Tissue transglutaminase (TGase 2) is proposed to be important for biomaterial-tissue interactions due to its presence and versatile functions in the extracellular environment. TGase 2 catalyzes the cross-linking of proteins through its Ca2+-dependent acyltransferase activity. Moreover, it enhances the interactions between fibronectin and integrins, which in turn mediates the adhesion, migration, and motility of the cells. TGase 2 is also a key player in the pathogenesis of fibrosis. In this study, we investigated whether TGase 2 is present at the biomaterial-tissue interface and might serve as an informative biomarker for the visualization of tissue response toward gelatin-based biomaterials. Two differently cross-linked hydrogels were used, which were obtained by the reaction of gelatin with lysine diisocyanate ethyl ester. The overall expression of TGase 2 by endothelial cells, macrophages, and granulocytes was partly influenced by contact to the hydrogels or their degradation products, although no clear correlation was evidenced. In contrast, the secretion of TGase 2 differed remarkably between the different cells, indicating that it might be involved in the cellular reaction toward gelatin-based hydrogels. The hydrogels were implanted subcutaneously in immunocompetent, hairless SKH1-Elite mice. Ex vivo immunohistochemical analysis of tissue sections over 112 days revealed enhanced expression of TGase 2 around the hydrogels, in particular at days 14 and 21 post-implantation. The incorporation of fluorescently labeled cadaverine derivatives for the detection of active TGase 2 was in accordance with the results of the expression analysis. The presence of an irreversible inhibitor of TGase 2 led to attenuated incorporation of the cadaverines, which verified the catalytic action of TGase 2. Our in vitro and ex vivo results verified TGase 2 as a potential biomarker for tissue response toward gelatin-based hydrogels. In vivo, no TGase 2 activity was detectable, which is mainly attributed to the unfavorable physicochemical properties of the cadaverine probe used.


Pilot study to evaluate knowledge of person-centred care, before and after a skill development programme, in a cohort of preregistration pharmacists within a large London hospital.
Aperson-centred approach to healthcare encompasses personalised care, supporting the individual to recognise their strengths and promote their independence. Preregistration pharmacists , who arenew pharmacy graduates undertaking their training year of practice before qualification,may need support embedding this in practice. To explore the knowledge and confidence in person-centred care (PCC) in a cohort of 12 preregistration pharmacists before and after receipt of an inhouse pharmacy-focused skill development programme using qualitative and quantitative measures. Two half-day skill development sessions were delivered to 12 preregistration pharmacists. Assessment forms were completed before and after the skill development sessions, including quantitative data gathered via Likert scales and qualitative, narrative responses. Responses were coded and classified into themes. Participants submitted one written reflective account demonstrating the use of PCC in medicines-related patient consultations following completion of the skill development programme. Assessment forms were received from all participants. Self-reported knowledge of PCC improved from the start of the first session to the end of the second session. The average score for the understanding of PCC rose from 6.5 to 9.6 (Likert scale of 0-10) to the end of the second session. Qualitative analysis identified five person-centred themes, including active listening, using open questions, supporting and empowering patients, developing a shared agenda, and encouraging patients to take ownership of their medicines. Confidence and knowledge increased from the start of session 1 to the end of session 2. and analysis of the reflective accounts and themes also identified increased knowledge of PCC. This work aligns with previous hospital trust work, which identified that a training programme increased awareness and provided foundation knowledge. A short programme to develop PCC skills was effective in improving the PCC knowledge of 12 preregistration pharmacists. A review of pharmacists' written accounts of their consultations suggests that these skills were integrated into practice.


[Radiotherapy of Hodgkin's disease].
After an historical review of the progressive improvement of Radiotherapy of Hodgkin's Disease, the authors present the actual management concepts with special reference to prognostic factors. So, the actual strategy of Hodgkin's Disease aims above all not to lower the "cure" values lately reached but to reduce the iatrogenic pathology trying not to treat to much the good prognosis subgroups. These are susceptible of a less aggressive therapy by the knowledge of the relation cost/benefit of the intensive radiotherapy and the several multidrug regimens.


PET and PET/CT with radiolabeled choline in prostate cancer: a critical reappraisal of 20 years of clinical studies.
We here aim to provide a comprehensive and critical review of the literature concerning the clinical applications of positron emission tomography/computed tomography (PET/CT) with radiolabeled choline in patients with prostate cancer (PCa). We will initially briefly summarize the historical context that brought to the synthesis of [11C]choline, which occurred exactly 20 years ago. We have arbitrarily grouped the clinical studies in three different periods, according to the year in which they were published and according to their relation with their applications in urology, radiotherapy and oncology. Studies at initial staging and, more extensively, studies in patients with biochemical failure, as well as factors predicting positive PET/CT will be reviewed. The capability of PET/CT with radiolabeled choline to provide prognostic information on PCa-specific survival will also be examined. The last sections will be devoted to the use of radiolabeled choline for monitoring the response to androgen deprivation therapy, radiotherapy, and chemotherapy. The accuracy and the limits of the technique will be discussed according to the information available from standard validation processes, including biopsy or histology. The clinical impact of the technique will be discussed on the basis of changes induced in the management of patients and in the evaluation of the response to therapy. Current indications to PET/CT, as officially endorsed by guidelines, or as routinely performed in the clinical practice will be illustrated. Emphasis will be made on methodological factors that might have influenced the results of the studies or their interpretation. Finally, we will briefly highlight the potential role of positron emission tomography/magnetic resonance and of new radiotracers for PCa imaging.


[Adsorption of insulin directly added to infusion bottles].
A study of insulin adsorption to infusion system was undertaken in the situation of direct addition of insulin alone in the infusion bottles, with use of 125I-labelled insulin and non labelled insulin. The infusion solutions tested were glucose solutions (5, 10, 15 and 30%). There is an important adsorption of insulin (75%) to the infusion system when the insulin concentration in the solution is low. The effluent insulin is not degraded. The glucose content of the solution does not influence the phenomenon. Adsorption of insulin decreases when the insulin concentration in the solution is increased and a plateau is reached starting from an insulin concentration in the solution of about 400 ng/ml; at higher concentrations, 80 to 90% of insulin that has been initially put in the bottle is recovered. Since the insulin concentrations in the solutions used in clinical practice are higher than 400 ng/ml, this adsorption phenomenon can be neglected and therefore can explain the efficiency of this mode of insulin-therapy. Kinetics of effluent insulin is a plateau, stable all along the infusion. Thus, direct addition of insulin alone into infusion bottles is an efficient and controlled mode of insulin-therapy if a minimal insulin concentration in the infused solution has been reached.


Design and application of a confocal microscope for spectrally resolved anisotropy imaging.
Biophysical imaging tools exploit several properties of fluorescence to map cellular biochemistry. However, the engineering of a cost-effective and user-friendly detection system for sensing the diverse properties of fluorescence is a difficult challenge. Here, we present a novel architecture for a spectrograph that permits integrated characterization of excitation, emission and fluorescence anisotropy spectra in a quantitative and efficient manner. This sensing platform achieves excellent versatility of use at comparatively low costs. We demonstrate the novel optical design with example images of plant cells and of mammalian cells expressing fluorescent proteins undergoing energy transfer.


Initial clinical assessment of a novel wound management system: a case series.
The importance of exudate management for maintaining local moisture balance and avoiding maceration in the chronic wound environment is well established. The authors performed the initial clinical testing of a novel wound management system, Sepaderm (Aalnex, Inc, Irvine, California), designed to vertically wick and sequester excess exudate away from wound/periwound tissues to promote a healthy wound environment. In this series of 14 patients with lower-extremity chronic venous leg and diabetic foot ulcers, the 3-component system was well tolerated and demonstrated the ability to prevent exudate leakage onto periwound tissue and reduce existing pain and itching. All ulcers lasting 1.2 to 360 months were previously treated with standard therapies, including human cell-derived skin substitutes in some of the patients. After treatment with the new system for 7 to 174 days, 8 patients had various degrees of wound closure, ranging from 44% to 100%. The 6 patients who failed to show wound closure were treated with the new system for an average of 5.7 days, but demonstrated other clinical benefits. Future studies in larger patient populations with quantitative wound closure assessments, as well as measurements of exudate, periwound maceration, and pain management, are needed.


Respiratory mucosal immunization with adenovirus gene transfer vector induces helper CD4 T cell-independent protective immunity.
Virus-vectored vaccine is a powerful activator of CD8 T cell-mediated immunity and is especially amenable to respiratory mucosal immunization, offering hopes for use in humans with diminished helper CD4 T cell function. However, whether virus-mediated mucosal immunization can produce immune protective CD8 T cells without the CD4 T cell help remains to be investigated. We used a replication-deficient adenovirus vector expressing an Mycobacterium tuberculosis antigen Ag85A for intranasal vaccination and evaluated its effect on CD8 T cell activation and protection in mice depleted of CD4 T cells. Intranasal vaccination of CD4 T cell-depleted mice led to suboptimal generation of Ag-specific tetramer(+) or interferon (IFN)-gamma-producing CD8 T cells in the lung and spleen but this was observed mainly at the early time after vaccination. Reduced CD8 T cell priming was also accompanied by decreased CD8 T cell responses (CTL). Nevertheless, the ratio of Ag-specific CD8 T cells to IFN-gamma-producing CD8 T cells in CD4 T cell-depleted hosts remained comparable to that in CD4 T cell-competent hosts. Furthermore, the 'unhelped' CD8 T cells also displayed a similar immune phenotype as the 'helped' counterparts. The animals with 'unhelped' CD8 T cells were as well-protected from pulmonary M. tuberculosis challenge as those with 'helped' CD8 T cells in the absence of CD4 T cells. The data obtained in the present study suggest that the fully immune protective CD8 T cells can still be generated by respiratory mucosal viral-mediated immunization without CD4 T cells and that CD8 T cells, 'helped' or 'unhelped', can confer significant protection against pulmonary tuberculosis independent of CD4 T cells.


Nef decreases HIV-1 sensitivity to neutralizing antibodies that target the membrane-proximal external region of TMgp41.
Primate lentivirus nef is required for sustained virus replication in vivo and accelerated progression to AIDS. While exploring the mechanism by which Nef increases the infectivity of cell-free virions, we investigated a functional link between Nef and Env. Since we failed to detect an effect of Nef on the quantity of virion-associated Env, we searched for qualitative changes by examining whether Nef alters HIV-1 sensitivity to agents that target distinct features of Env. Nef conferred as much as 50-fold resistance to 2F5 and 4E10, two potent neutralizing monoclonal antibodies (nAbs) that target the membrane proximal external region (MPER) of TMgp41. In contrast, Nef had no effect on HIV-1 neutralization by MPER-specific nAb Z13e1, by the peptide inhibitor T20, nor by a panel of nAbs and other reagents targeting gp120. Resistance to neutralization by 2F5 and 4E10 was observed with Nef from a diverse range of HIV-1 and SIV isolates, as well as with HIV-1 virions bearing Env from CCR5- and CXCR4-tropic viruses, clade B and C viruses, or primary isolates. Functional analysis of a panel of Nef mutants revealed that this activity requires Nef myristoylation but that it is genetically separable from other Nef functions such as the ability to enhance virus infectivity and to downregulate CD4. Glycosylated-Gag from MoMLV substituted for Nef in conferring resistance to 2F5 and 4E10, indicating that this activity is conserved in a retrovirus that does not encode Nef. Given the reported membrane-dependence of MPER-recognition by 2F5 and 4E10, in contrast to the membrane-independence of Z13e1, the data here is consistent with a model in which Nef alters MPER recognition in the context of the virion membrane. Indeed, Nef and Glycosylated-Gag decreased the efficiency of virion capture by 2F5 and 4E10, but not by other nAbs. These studies demonstrate that Nef protects lentiviruses from one of the most broadly-acting classes of neutralizing antibodies. This newly discovered activity for Nef has important implications for anti-HIV-1 immunity and AIDS pathogenesis.


Infection of macrophage-like THP-1 cells with Mycobacterium avium results in a decrease in their ability to phosphorylate nucleolin.
This study of the phosphorylation ability of macrophage-like cells upon infection with Mycobacterium avium was undertaken to establish potential targets of the interference with host response mechanisms. Cytosolic and membrane fractions from noninfected and infected cells were incubated with [gamma-(32)P]ATP, in the presence of Mg(2+) and the absence of Ca(2+), and the patterns of phosphoproteins synthesized were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Lower levels of a 110-kDa phosphoprotein were observed in association with cytosolic fractions from mycobacterium-infected cells compared to noninfected cells or cells treated with lipopolysaccharide or having ingested Escherichia coli or killed M. avium. The 110-kDa phosphoprotein was present in the soluble fraction (230,000 x g supernatant) after the kinase incubation, from where it was partially purified and identified as phosphonucleolin by amino acid sequencing. The decrease in nucleolin phosphorylation observed was not related to changes in the cytosolic or membrane levels of this protein, and was detected also in the cytosolic fraction of (32)P-labeled intact cells.


Expert consensus: time for a change in the way we advise our patients to use topical corticosteroids.
Topical corticosteroids form the mainstay of treatment for many skin conditions. If used appropriately, they are safe and effective, and side-effects are generally uncommon. Current advice to patients to apply topical corticosteroid preparations 'sparingly' or 'thinly' contributes to 'steroid phobia', increasing the risk of poor clinical response and treatment failure. Such cautionary advice also overlooks the fact that the vast majority of patients are prescribed topical corticosteroids of mild potency for which the evidence suggests that the risk of harm is minimal. In the patient's mind, the current advice groups all steroids together regardless of their potential for adverse effects. The advice also tends to reinforce an erroneous concern that the risks from topical corticosteroids may be similar to those from systemic corticosteroids. We propose a change to make the pharmacy labelling of topical corticosteroids more accurately reflect the low risk of harm from corticosteroids of low to moderate potency and the importance of applying sufficient medication to achieve a satisfactory clinical response. This change could provide the focus for updated, evidence-based education for healthcare professionals in prescribing of topical corticosteroids and help in the provision of more appropriate advice to patients. We recommend that patients are informed that treatment should not exceed prescribed quantities, and continuing treatment should be under careful medical supervision. We also recommend that topical corticosteroid products include clear 'fingertip unit' instructions, preferably with images of a 'fingertip unit' and a chart to show the number of units required for specific areas of the body.


Utilization of Echocardiogram, Carotid Ultrasound, and Cranial Imaging in the Inpatient Investigation of Syncope: Its Impact on the Diagnosis and the Patient's Length of Hospitalization.
Although guidelines suggest that the best strategy for evaluating syncope is clinical history and physical examination, the inappropriate utilization of diagnostic imaging is common. A single center retrospective analysis conducted in adult patients admitted for evaluation and management of syncope for a period of 12 months. Charts were reviewed to abstract demographic data, admitting and discharge diagnosis, diagnostic investigatory tests including imaging modalities (echocardiogram, carotid ultrasound, and cranial computed tomography (CT)) ordered, subspecialty consultation requested, treatment rendered and hospital length of stay (LOS). A total of 109 patients were admitted for syncope, mean age was 68.74 ± 21.04 years and 39.44% were men. Echocardiogram, carotid ultrasound, and cranial CT were ordered in 69.72%, 33.02%, and 76.14% respectively. The mean hospital LOS was 2.6 days. Patients with no imaging test, one imaging test, two imaging tests, and three imaging tests ordered have an average hospital LOS of 2.22 days, 2.44 days, 2.58 days, and 3.07 days respectively. The number of imaging test and its relation to the admitting (Chi-square (chi-sq) P = 0.4165, nominal logistic regression (LR) P = 0.939) and discharge (chi-sq P = 0.1507, nominal LR P = 0.782) diagnosis as well as the LOS in relation to the number of imaging test ordered (analysis of variance (ANOVA) P = 0.368, Kruskal Wallis (KW) P = 0.352) were not statistically significant although there was a trend of prolonged hospital LOS the more imaging diagnostic test had been ordered. Syncope was the admitting and discharge diagnosis in 89.9% and 91.74% respectively. Choosing the appropriate diagnostic tests as dictated by the patient's clinical manifestation and utilizing less expensive test would be appropriate and cost-effective approach in appraising patients with syncope.


Antimicrobial Usages and Antimicrobial Resistance in Commensal Escherichia coli From Veal Calves in France: Evolution During the Fattening Process.
Extended-Spectrum-Cephalosporin (ESC)-resistant Enterobacteriaceae have widely spread in all settings worldwide. In animals, Extended-Spectrum Beta-Lactamase (ESBL) producers have been frequently identified in veal calves. The objectives of this study were to investigate the trends in the ESBL load and antimicrobial resistance (AMR) proportions, and antimicrobial usages (AMU) in veal calves during the fattening process. Ten fattening farms were selected and 50 animals per farm were sampled. AMR was assessed in bacteria from the dominant flora (collected on non-selective MacConckey agar) and in ESBL/AmpC-carrying bacteria from the subdominant flora (selected on ChromID ESBL selective plates) upon arrival and 5-6 months later before slaughter. The number and types of treatments during fattening were also collected. Rates of ESBL-producing E. coli from the subdominant flora significantly decreased in all farms (arrival: 67.7%; departure: 20.4%) whereas rates of multidrug-resistant E. coli from the dominant flora have significantly increased (arrival: 60.2%; departure: 67.2%; p = 0.025). CTX-M-1 was the most frequently identified ESBL enzyme (arrival: 59.3%; departure: 52.0%). The plasmid-mediated mcr-1 gene was also identified occasionally. In parallel, levels of resistances to non-critically important antimicrobials were already high upon arrival but have still further increased over time until slaughter. Our study also highlighted that if only ESBL-producing isolates were monitored, it might have led to a partial (and partly false) picture of AMR rates globally decreasing during the fattening period. The mean number of antimicrobial treatments per calf (NTPC) was 8.75 but no association between AMU and AMR was evidenced. Most ESBL producers were clonally unrelated suggesting multiple sources and not cross-contaminations among calves during transportation. Feeding milk containing antimicrobial residues to veal calves is hypothesized to explain the high ESBL loads in animals at the entrance on farms.


The protonmotive potential difference across the vacuo-lysosomal membrane of Hevea brasiliensis (rubber tree) and its modification by a membrane-bound adenosine triphosphatase.
The vacuo-lysosomes of Hevea brasiliensis (rubber tree) constitute a suitable model system for the study of active transport and energization at the level of the membrane of plant vacuoles. The pH gradient (delta pH) and the membrane potential (delta psi) of vacuo-lysosomes were determined by means of the weak base methylamine and the lipophilic cation tetraphenylphosphonium. The values obtained depended strongly on the experimental conditions such as medium pH or K+ concentration. Under experimental conditions, i.e., pH 7.5 outside and low K+, the delta pH amounts to about 0.9 unit, interior acid, and the delta psi to -120 mV, interior negative. The delta psi is presumably caused by the imposed K+ gradient, and the internal acidification might be a consequence of the passive proton inflow along the electric field. This explanation is sustained by the ineffectiveness of carbonyl cyanide p-trifluoromethoxyphenylhydrazone in destroying the delta pH and delta psi, whereas higher K+ concentration decreased both. Under conditions existing in vivo, the membrane potential might be significantly lower. The presence of ATP increased the acidification of the intravesicular space by 0.5pH unit to a delta pH of up to 1.4 and shifts the membrane potential at least 60mV to a more positive value. The change of the protonmotive potential did not occur with ADP; the pH-dependence of the change was identical with the pH-dependence of a vacuo-lysosomal membrane-bound ATPase, and the effect of ATPase was prevented by the presence of the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone. The change of protonmotive potential difference, brought about by the ATPase, was at least 90 mV. This is evidence that a vacuo-lysosomal ATPase in plants can function as an electrogenic proton pump that transfers protons into the vacuo-lysosomal space.


Immunity to Hammondia hammondi infection in cats.
Acquisition of immunity to Hammondia hammondi, a newly recognized coccidian of cats, was studied in 18 specific-pathogen-free cats. One cat was given a single oral inoculation, 11 cats were given 2 oral inoculations, and 1 cat was given 3 oral inoculations of homogenized mouse carcasses containing H hammondi. In all cats, oocyst shedding began 6 to 9 days after the 1st inoculation. Oocyst shedding peaked at 1 to 2 days after the onset of shedding and lasted for 1 to 2 weeks. None of the cats became sick. Of the 11 cats inoculated twice (between 2-51 days after the 1st inoculation), 5 shed oocysts 7 to 14 days after the repeat inoculation; however, fewer oocysts were shed at this time. One cat that was inoculated thrice (14 and 51 days after the 1st inoculation) shed oocysts 14 to 17 days after the 3rd inoculation but not after the 2nd inoculation. Spontaneous oocyst shedding was studied in 9 of these 13 H hammondi-infected cats for 5 months. Two cats spontaneously shed oocysts: One cat (inoculated only once) spontaneously re-shed oocysts 21 to 24, 31 to 33, 49 to 50, and 118 to 120 days after inoculation; The other cat (inoculated twice-the 2nd time, 6 days after the 1st inoculation) re-shed oocysts 38 to 48, 85 to 89, and 133 to 136 days after the 1st inoculation. The course of H hammondi infection was studied in 5 cats given weekly injections of 6-methyl prednisolone acetate for at least 7 weeks, starting 18 days before inoculation in 2 cats, and starting 14, 34, and 45 day after inoculation in 3 cats. The induced hyperadrenocorticism did not affect the prepatent period or induce parasitism of extraintestinal organs. The 3 cats infected for 14, 34, and 45 days, re-shed oocysts after hyperadrenocorticism was induced. It was concluded that immunity to H hammondi infection in cats is less stable than immunity to the related coccidian, Toxoplasma gondii.


Spinal Anesthesia Reduces Myocardial Ischemia-triggered Ventricular Arrhythmias by Suppressing Spinal Cord Neuronal Network Interactions in Pigs.
Cardiac sympathoexcitation leads to ventricular arrhythmias. Spinal anesthesia modulates sympathetic output and can be cardioprotective. However, its effect on the cardio-spinal reflexes and network interactions in the dorsal horn cardiac afferent neurons and the intermediolateral nucleus sympathetic neurons that regulate sympathetic output is not known. The authors hypothesize that spinal bupivacaine reduces cardiac neuronal firing and network interactions in the dorsal horn-dorsal horn and dorsal horn-intermediolateral nucleus that produce sympathoexcitation during myocardial ischemia, attenuating ventricular arrhythmogenesis. Extracellular neuronal signals from the dorsal horn and intermediolateral nucleus neurons were simultaneously recorded in Yorkshire pigs (n = 9) using a 64-channel high-density penetrating microarray electrode inserted at the T2 spinal cord. Dorsal horn and intermediolateral nucleus neural interactions and known markers of cardiac arrhythmogenesis were evaluated during myocardial ischemia and cardiac load-dependent perturbations with intrathecal bupivacaine. Cardiac spinal neurons were identified based on their response to myocardial ischemia and cardiac load-dependent perturbations. Spinal bupivacaine did not change the basal activity of cardiac neurons in the dorsal horn or intermediolateral nucleus. After bupivacaine administration, the percentage of cardiac neurons that increased their activity in response to myocardial ischemia was decreased. Myocardial ischemia and cardiac load-dependent stress increased the short-term interactions between the dorsal horn and dorsal horn (324 to 931 correlated pairs out of 1,189 pairs, P < 0.0001), and dorsal horn and intermediolateral nucleus neurons (11 to 69 correlated pairs out of 1,135 pairs, P < 0.0001). Bupivacaine reduced this network response and augmentation in the interactions between dorsal horn-dorsal horn (931 to 38 correlated pairs out of 1,189 pairs, P < 0.0001) and intermediolateral nucleus-dorsal horn neurons (69 to 1 correlated pairs out of 1,135 pairs, P < 0.0001). Spinal bupivacaine reduced shortening of ventricular activation recovery interval and dispersion of repolarization, with decreased ventricular arrhythmogenesis during acute ischemia. Spinal anesthesia reduces network interactions between dorsal horn-dorsal horn and dorsal horn-intermediolateral nucleus cardiac neurons in the spinal cord during myocardial ischemia. Blocking short-term coordination between local afferent-efferent cardiac neurons in the spinal cord contributes to a decrease in cardiac sympathoexcitation and reduction of ventricular arrhythmogenesis.


Parenting desires, parenting intentions, and anticipation of stigma upon parenthood among lesbian, bisexual, and heterosexual women in Portugal.
We explored parenting desires, parenting intentions, and anticipation of stigma upon parenthood in a sample of 257 self-identified lesbian, bisexual, and heterosexual childfree women in Portugal. No differences between the groups were observed concerning parenting desires. However, lesbian and bisexual women reported lower intentions to have children than their heterosexual counterparts. Although lesbian women considered themselves to have a higher chance of being victims of social stigma as mothers, this was not associated with their parenting desires. Furthermore, younger lesbian women intended to have children to a greater extent than did older lesbian women; relational status did not relate to lesbian women's parenting intentions, desires, or anticipation of stigma. Overall, this study contributes to knowledge about family formation processes among Portuguese women diverse in sexual identity.


Crystal structure-based selective targeting of the pyridoxal 5'-phosphate dependent enzyme kynurenine aminotransferase II for cognitive enhancement.
Fluctuations in the brain levels of the neuromodulator kynurenic acid may control cognitive processes and play a causative role in several catastrophic brain diseases. Elimination of the pyridoxal 5'-phosphate dependent enzyme kynurenine aminotransferase II reduces cerebral kynurenic acid synthesis and has procognitive effects. The present description of the crystal structure of human kynurenine aminotransferase II in complex with its potent and specific primary amine-bearing fluoroquinolone inhibitor (S)-(-)-9-(4-aminopiperazin-1-yl)-8-fluoro-3-methyl-6-oxo-2,3-dihydro-6H-1-oxa-3a-azaphenalene-5-carboxylic acid (BFF-122) should facilitate the structure-based development of cognition-enhancing drugs. From a medicinal chemistry perspective our results demonstrate that the issue of inhibitor specificity for highly conserved PLP-dependent enzymes could be successfully addressed.


Cartilage oligomeric matrix protein associates with granulin-epithelin precursor (GEP) and potentiates GEP-stimulated chondrocyte proliferation.
Mutations in human cartilage oligomeric matrix protein (COMP) have been linked to the development of pseudoachondroplasia and multiple epiphyseal dysplasia; however, the functions of both wild-type and mutant COMP in the skeletogenesis remain unknown. In an effort to define the biological functions of COMP, a functional genetic screen based on the yeast two-hybrid system was performed. This led to the identification of granulin-epithelin precursor (GEP), an autocrine growth factor, as a COMP-associated partner. COMP directly binds to GEP both in vitro and in vivo, as revealed by in vitro pull down and co-immunoprecipitation assays. GEP selectively interacts with the epidermal growth factor repeat domain of COMP but not with the other three functional domains of COMP. The granulin A repeat unit of GEP is required and sufficient for association with COMP. COMP co-localizes with GEP predominantly in the pericellular matrix of transfected rat chondrosarcoma cell and primary human chondrocytes. Staining of musculoskeletal tissues of day 19 mouse embryo with antibodies to GEP is restricted to chondrocytes in the lower proliferative and upper hypertrophic zones. Overexpression of GEP stimulates the proliferation of chondrocytes, and this stimulation is enhanced by COMP. In addition, COMP appears to be required for GEP-mediated chondrocyte proliferation, since chondrocyte proliferation induced by GEP is dramatically inhibited by an anti-COMP antibody. These findings provide the first evidence linking the association of COMP and GEP and identifying a previously unrecognized growth factor (i.e. GEP) in cartilage.


Modeling the mechanisms of biological GTP hydrolysis.
Enzymes that hydrolyze GTP are currently in the spotlight, due to their molecular switch mechanism that controls many cellular processes. One of the best-known classes of these enzymes are small GTPases such as members of the Ras superfamily, which catalyze the hydrolysis of the γ-phosphate bond in GTP. In addition, the availability of an increasing number of crystal structures of translational GTPases such as EF-Tu and EF-G have made it possible to probe the molecular details of GTP hydrolysis on the ribosome. However, despite a wealth of biochemical, structural and computational data, the way in which GTP hydrolysis is activated and regulated is still a controversial topic and well-designed simulations can play an important role in resolving and rationalizing the experimental data. In this review, we discuss the contributions of computational biology to our understanding of GTP hydrolysis on the ribosome and in small GTPases.


The immunophilin-interacting protein AtFIP37 from Arabidopsis is essential for plant development and is involved in trichome endoreduplication.
The FKBP12 (FK506-binding protein 12 kD) immunophilin interacts with several protein partners in mammals and is a physiological regulator of the cell cycle. In Arabidopsis, only one specific partner of AtFKBP12, namely AtFIP37 (FKBP12 interacting protein 37 kD), has been identified but its function in plant development is not known. We present here the functional analysis of AtFIP37 in Arabidopsis. Knockout mutants of AtFIP37 show an embryo-lethal phenotype that is caused by a strong delay in endosperm development and embryo arrest. AtFIP37 promoter::beta-glucuronidase reporter gene constructs show that the gene is expressed during embryogenesis and throughout plant development, in undifferentiating cells such as meristem or embryonic cells as well as highly differentiating cells such as trichomes. A translational fusion with the enhanced yellow fluorescent protein indicates that AtFIP37 is a nuclear protein localized in multiple subnuclear foci that show a speckled distribution pattern. Overexpression of AtFIP37 in transgenic lines induces the formation of large trichome cells with up to six branches. These large trichomes have a DNA content up to 256C, implying that these cells have undergone extra rounds of endoreduplication. Altogether, these data show that AtFIP37 is critical for life in Arabidopsis and implies a role for AtFIP37 in the regulation of the cell cycle as shown for FKBP12 and TOR (target of rapamycin) in mammals.


SARS-CoV-2 seroprevalence determination in pets and camels in Egypt using multispecies enzyme-linked immunosorbent assay.
The coronavirus disease 2019 (COVID-19) pandemic has translated into a worldwide economic recession and public health crisis. Bats have been incriminated as the main natural host for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the causative agent of the COVID-19 pandemic. However, the reservoir and carrier hosts of the virus remain unknown. Therefore, a cross sectional serosurvey study was performed to estimate antibodies to SARS-CoV-2. To assess IgM antibodies to SARS-CoV-2 nucleocapsid protein (NP), a SARS-CoV-2 Double Antigen Multispecies diagnostic enzyme-linked immunosorbent assay kit was used. The seropositive samples were confirmed and validated by measuring IgG antibody titers in sera. The enrolled animals were from different locations in the Giza governorate, Egypt, and were sampled at the time of the pandemic; they comprised 92 companion animals and 92 domestic camels. The study established that 4.76% (1/21 clinical samples) of dogs, 7.69% of cats (1/13 shelter samples) and 1.08% (1/92) of camels, had measurable SARS-CoV-2 NP IgM antibodies. All IgM-seropositive samples were IgG positive with a measurable titer of 34.5, 28.6, and 25.8 UI/mL for dog, cat, and camels, respectively. According to our best knowledge, this study was the first to assess SARS-CoV-2 seroprevalence in the specific animals investigated in Egypt. These results may herald a promising epidemiological role for pet animals and camels in SARS-CoV-2 virus maintenance. Thus, our study's results ought to be confirmed with a nationwide seroprevalence study, and further studies are required to clarify whether these animals act as active or passive carriers.


[Nursing care to patient on electroconvulsive therapy].
Eletric convulsive shock therapy has become a controversial topic in psychiatric services. Patients continue to have electroconvulsive therapy. The team nursing has seen the treatment as just another task. In this paper the authors present how the nurse can help the patient who is submitted to electroconvulsive therapy.


Assessment of Black Patients' Perception of Their Dermatology Care.
The availability and quality of skin and hair care for black patients in the United States has been a subject of growing interest in dermatology. There is limited understanding of the perceptions of black patients about the care they receive from dermatologists and the factors affecting their care satisfaction. To elucidate black patients' perceptions of their dermatology experience in and outside of a skin of color clinic (SOCC). A cross-sectional study involving a survey and focus groups was conducted including adult black patients treated in an SOCC. Recruitment was conducted April through June 2015 through clinic-posted flyers. Participants engaged in 1 of 4 focus groups on July 14, 15, 29, or 30, 2015, and completed a survey. Data were analyzed March 2016 through June 2017. Planned outcomes of the focus groups were black patients' inductively generated themes on their perspectives and experiences in dermatology clinics. Planned outcomes of the survey were patients' ratings of SOCC and non-SOCC dermatologists in terms of interaction style, cultural awareness, and treatment satisfaction. Importance of patient-dermatologist racial concordance was a planned outcome in both measures. Given lack of existing prior research, no specific hypotheses were generated. Of the 19 adult black patients who participated in the study, 18 (95%) were women, and the mean (SD) age was 50.0 (14.2) years. All patients reported positive experiences in the SOCC. Compared with non-SOCC dermatology treatment experiences, patients reported higher levels of overall satisfaction with SOCC dermatologists (t13 = 2.85; P = .01). Patients perceived that SOCC dermatologists were better trained to care for black patients (t13 = 4.42; P = .001); showed patients greater respect and dignity (t13 = 3.37; P = .005), as well as understanding (t13 = 2.56, P = .02); and were more trustworthy (t13 = 3.47; P = .004). The majority of the comments in the focus groups (n = 207) described 2 themes: dermatologists' interaction style (62/207; 30.0%) and knowledge about black skin and hair (42/207; 20.3%). Other themes were partnering with patients on outcomes (17/207; 8.2%), shared life experiences (14/207; 6.8%), and economic sensitivity (7/207; 3.4%). These themes accounted for a large part of the participants' discussion. Of all respondents, 71% (12/17) stated that they would prefer a black (or race concordant) dermatologist; this included 91% (10/11) of the race-concordant group and 33% (2/6) of the race-discordant group. Participants reported that the SOCC dermatologists provided unique and uniformly beneficial care to black patients. Care satisfaction appeared most related to dermatologists' interpersonal style and specialized knowledge in care of black skin and hair. Findings suggest that black patients' dermatologic care satisfaction would increase if dermatologists underwent enhanced residency training in skin of color, cultural competency, cost-conscious care, and empathic communication skills, and if there were greater dermatology workforce diversity.


[Functional somatic diseases--a review].
Functional somatic illness is a clinical concept used to define medically unexplained somatic symptoms considered to express psychological distress. Functional somatic illness may express underlying psychiatric disorders (e.g. fibromyalgia due to non-fearful panic disorder, irritable bowel syndrome due to bipolar disorder). Sustained physiological activation caused by stressful life events combined with catastrophic thinking may be another cause. Functional somatic illness may also be caused by classic conditioning of physiological responses that may have been triggered by biological or emotional stimuli. Operant conditioning may also be a cause. The therapeutic alliance relies on acceptance of the reality of the subjective complaints, without a priori acceptance of the patient's attribution of the cause of the symptoms. We recommend initial exploration of the patient's own ideas about aetiology, including appropriate medical tests. The physician should then change the agenda to a biopsychosocial perspective and identify current stressors and psychosocial variables that reinforce symptoms. Only a few randomised trials have been performed. They suggest that psychological treatment should be systematic and structured, with a focus on information, alternative ways of perception, and problem solving. Active forms of physiotherapy and psychopharmacological drugs may be of some benefit in selected patients.


Clinical Barriers to Effective Treatment of Eating Disorders and Co-occurring Psychiatric Disorders in Transgendered Individuals.
Eating disorders (EDs) are complex and difficult to treat illnesses that are often chronic and disabling on their own accord or due to comorbidity with other psychiatric conditions. Historically, EDs have been viewed as illnesses of heterosexual, affluent white females. This stereotype increases the likelihood that these disorders will be underrecognized in the lesbian, gay, bisexual, and transgender community, as well as in different gender, socioeconomic, and ethnic populations. Our case report illustrates the clinical difficulties of managing a patient who presented to inpatient treatment with complaints of depression and suicidality, but who also had an active ED and was transgender.


Synthesis of porous upconverting luminescence α-NaYF4:Ln(3+) microspheres and their potential applications as carriers.
The present work reports a self-sacrificing template strategy to synthesize porous α-NaYF4 microspheres via the reaction of as-prepared Y(OH)CO3·H2O@SiO2 with NH4F and NaNO3 solutions. XRD, SEM, TEM and N2 adsorption-desorption measurements were used to characterize the resulting product. The surface SiO2 shell was shown to play a vital role in size and shape control and porosity formation. A possible reaction mechanism was explored in terms of a surface-protected etching and ion-exchange reaction process. To explore their application potential, the storage and release behavior of Rhodamine 640 dye in the porous α-NaYF4 microspheres was investigated, showing a relatively high loading efficiency and a sustained release ratio. Under near-infrared (NIR) irradiation, porous α-NaYF4 microspheres doped with lanthanide ions showed typical upconverting luminescence characteristics that can convert NIR photons to ultraviolet/visible photons. The above features and properties indicate that our present porous upconverting luminescence particles are promising in biological applications as luminescence imaging agents and drug carriers.


[Ruptured cerebral aneurysm of the median artery of the corpus callosum (accessory anterior cerebral artery): case report].
A rare case of ruptured cerebral aneurysm of median artery of corpus callosum (accessory anterior cerebral artery: Acc ACA) is reported. A 66-year-old hypertensive female suddenly lost consciousness for 5 minutes and later complained of severe headache. On admission, the next day, consciousness was clear and she showed no neurological deficit except for right motor weakness. CT scan revealed subarachnoid hemorrhage, and carotid angiography showed triplicated anterior cerebral artery with a saccular aneurysm on the Acc ACA. Twenty hours after the onset, an operation was performed to clip the aneurysm neck. Post-operative course was uneventful and she was discharged on the 25th postoperative day without neurological deficits. When the median artery of corpus callosum(MACC), a branch of anterior communicating artery distributes to one or to both hemispheres, it is called Acc ACA. It is thought to be a vascular anomaly and which has an incidence of 20%. However, cases of aneurysm of MACC (or Acc ACA) have not been reported and our case is considered to be the first.


Competitive NMDA receptor antagonists and spinal-cord ischemia.
Ischemic neuronal death is associated with excitatory amino acid (EAA) release. Their action is mediated by N-methyl-D-aspartate (NMDA) receptors. Blockade of the receptors before the ischemic insult can decrease neuronal damage. Accordingly, we investigated the protective effect during spinal cord ischemia of two competitive antagonists, 4-(3-phosphonopropyl)-2-piperazine-carboxylic acid (CPP) and cis-4-(phosphonomethyl)-2-piperidine-carboxylic acid (CGS). Male Sprague-Dawley rats underwent intrathecal administration of 10 microL saline, CGS, and CPP 10 mM solutions, in a randomized blinded fashion, and were subjected to balloon occlusion of the thoracic aorta. Proximal aortic pressure was lowered to a mean of 40 mm Hg by partial exsanguination. In the acute protocol, 21 rats divided in 3 groups of 7 (saline, CPP, and CGS) were used to calculate the aortic occlusion time (AOT) resulting in paraplegia in 50% of animals (P50). In the chronic study, 24 rats divided in 4 groups of 6 (saline, CPP, CGS, sham) underwent 12-min occlusion. The chronic animals were scored daily for 28 days and submitted to histology of the cord. In the acute study, the P50 of CGS (10 min 48 s) and CPP (11 min 11 s) was longer than saline (10 min 27 s). In the chronic groups, analysis of variance of neurologic (p = .66) and histologic (p = .66) scores did not disclose differences between CGS, CPP, and saline. In conclusion, blockade of NMDA receptors with CPP or CGS may afford some protection for durations of occlusion around the P50, but it is not beneficial when ischemic injury is more protracted.


Repertoire, unified nomenclature and evolution of the Type III effector gene set in the Ralstonia solanacearum species complex.
Ralstonia solanacearum is a soil-borne beta-proteobacterium that causes bacterial wilt disease in many food crops and is a major problem for agriculture in intertropical regions. R. solanacearum is a heterogeneous species, both phenotypically and genetically, and is considered as a species complex. Pathogenicity of R. solanacearum relies on the Type III secretion system that injects Type III effector (T3E) proteins into plant cells. T3E collectively perturb host cell processes and modulate plant immunity to enable bacterial infection. We provide the catalogue of T3E in the R. solanacearum species complex, as well as candidates in newly sequenced strains. 94 T3E orthologous groups were defined on phylogenetic bases and ordered using a uniform nomenclature. This curated T3E catalog is available on a public website and a bioinformatic pipeline has been designed to rapidly predict T3E genes in newly sequenced strains. Systematical analyses were performed to detect lateral T3E gene transfer events and identify T3E genes under positive selection. Our analyses also pinpoint the RipF translocon proteins as major discriminating determinants among the phylogenetic lineages. Establishment of T3E repertoires in strains representatives of the R. solanacearum biodiversity allowed determining a set of 22 T3E present in all the strains but provided no clues on host specificity determinants. The definition of a standardized nomenclature and the optimization of predictive tools will pave the way to understanding how variation of these repertoires is correlated to the diversification of this species complex and how they contribute to the different strain pathotypes.


RHC 3659: a new orally active angiotensin converting enzyme inhibitor in normal volunteers.
1 The new converting enzyme inhibitor RHC 3659 was tested in 15 male volunteers. The study consisted of two parts: first, the ability of a single oral dose (5, 10, 20, 40 or 80 mg) to inhibit the pressor response to exogenous angiotensin I was tested with blood pressure and heart rate monitored continuously through an intraarterial catheter. A dose-related shift to the right of the pressor response curve to angiotensin I was observed with a peak occurring within 0.5 to 1 h. The pressor response to angiotensin II was unaffected. 2 In the second part, plasma renin and converting enzyme activity, angiotensin II and aldosterone were measured serially before and up to 8 h after administration of a single oral dose of RHC 3659. As expected. plasma angiotensin II and aldosterone fell within 30 min while plasma renin activity increased. Plasma converting enzyme activity was suppressed at 0.5 h in a dose-related manner with levels still below 30% of control 4 h following 80 mg of the inhibitor. 3 However, in vitro the enzyme-inhibitor complex seemed quited fragile since during storage of the plasma samples at -20 degrees C, converting enzyme activity increased significantly already within days (P less than 0.001, n = 28) and continued to rise for more than 2 months. This fragility may explain the seemingly lower potency of RHC 3659 when compared to captopril. No side effects were observed.


Surface-immobilized dextran limits cell adhesion and spreading.
Dextran has recently been investigated as an alternative to polyethylene glycol (PEG) for low protein-binding, cell-resistant coatings on biomaterial surfaces. Although anti-fouling properties of surface-grafted dextran and PEG are quite similar, the multivalent properties of dextran are advantageous when high-density surface immobilization of biologically active molecules to low protein-binding surface coatings is desired. The preferred methods of dextran immobilization for biomaterial applications should be simple with minimal toxicity. In this report, a method is described for covalent immobilization of dextran to material surfaces which involves low residual toxicity reagents in mild aqueous reaction conditions. 70 kDa MW dextran was immobilized on glass and polyethylene terephthalate (PET) surfaces. 3T3 fibroblast cell adhesion was compared on untreated, aminated, and dextran-coated materials. Dextran coatings effectively limited cell adhesion and spreading on glass and PET surfaces in the presence of serum-borne cell adhesion proteins. With dextran-based surface coatings, it will be possible to develop well-defined surface modifications that promote specific cell interactions and perhaps better performance in long-term biomaterial implants.


Human cytomegalovirus IE86 protein binds to cellular Mcm3 protein but does not inhibit its binding to the Epstein-Barr virus oriP in U373MG-p220.2 cells.
Human cytomegalovirus (HCMV) or its immediate-early IE86 protein alone induces cell cycle in quiescent primary human foreskin fibroblasts (HFFs), but blocks its progression at the G1/S interphase and inhibits cellular DNA synthesis by a mechanism that is not clearly understood. It is assumed that, in this phenomenon, the binding of minichromosome maintenance (Mcm) proteins to replication origins is blocked. In this work, we analyzed the initiation of DNA replication in HCMV-permissive U373MG cells and used oriP of Epstein-Barr virus (EBV) as a simplified model of a cellular replication origin. Using U373MG cells we found that HCMV IE86 protein was bound to Mcm3, but did not inhibit the cellular DNA synthesis. Using U373MG-p220.2 cells carrying EBV oriP and expressing Epstein-Barr nuclear antigen 1 (EBNA1), we found that EBNA1 as well as Mcm3 were bound to oriP and that neither HCMV nor IE86 protein inhibited the binding of Mcm3 to oriP. Differences between the effects of HCMV on the cell cycle progression in HFFs and U373MG cells are discussed. cell cycle; Human cytomegalovirus; DNA replication.


Practical system for the generation of pulsed quantum frequency combs.
The on-chip generation of large and complex optical quantum states will enable low-cost and accessible advances for quantum technologies, such as secure communications and quantum computation. Integrated frequency combs are on-chip light sources with a broad spectrum of evenly-spaced frequency modes, commonly generated by four-wave mixing in optically-excited nonlinear micro-cavities, whose recent use for quantum state generation has provided a solution for scalable and multi-mode quantum light sources. Pulsed quantum frequency combs are of particular interest, since they allow the generation of single-frequency-mode photons, required for scaling state complexity towards, e.g., multi-photon states, and for quantum information applications. However, generation schemes for such pulsed combs have, to date, relied on micro-cavity excitation via lasers external to the sources, being neither versatile nor power-efficient, and impractical for scalable realizations of quantum technologies. Here, we introduce an actively-modulated, nested-cavity configuration that exploits the resonance pass-band characteristic of the micro-cavity to enable a mode-locked and energy-efficient excitation. We demonstrate that the scheme allows the generation of high-purity photons at large coincidence-to-accidental ratios (CAR). Furthermore, by increasing the repetition rate of the excitation field via harmonic mode-locking (i.e. driving the cavity modulation at harmonics of the fundamental repetition rate), we managed to increase the pair production rates (i.e. source efficiency), while maintaining a high CAR and photon purity. Our approach represents a significant step towards the realization of fully on-chip, stable, and versatile sources of pulsed quantum frequency combs, crucial for the development of accessible quantum technologies.


Therapists' experiences with providing guided internet-delivered cognitive behavioral therapy for patients with mild to moderate depression: a thematic analysis.
Guided internet-delivered therapy has shown promising results for patients with mild to moderate depressive disorder, but several challenges with the format have been reported. The aim of this qualitative study was to investigate therapists' experiences providing guided internet-delivered cognitive behavioral therapy for patients with mild to moderate depression. Twelve therapists were interviewed, and the interviews were analyzed using reflexive thematic analysis. Three themes were created: (1) For the right person, at the right time. This theme is about therapists' experiences appointing patients to the program. It is challenging to predict which patients will benefit from it, and it is not the right option for all patients. (2) It is not like chatting on Facebook. The second theme was about the experiences with demands on clinics, therapists and patients that must be considered. The internet-delivered treatment should not be viewed as a simple treatment option, and the value of having contact with the patients during treatment was emphasized. (3) It is like a railroad, but without the switches. This theme was about the experiences with how the treatment content was conveyed to the patients, how the therapists expressed concerns with the usability of the program and the reported need for more possibilities in tailoring treatment for each patient.


Changes in bronchial epithelium in relation to cigarette smoking, 1955-1960 vs. 1970-1977.
To test the hypothesis that the reduction in tar and nicotine content of cigarette smoke that began in the 1950s should be reflected by the histologic changes in the bronchial epithelium of cigarette smokers, 20, 424 sections taken at autopsy from the bronchial tubes of 445 men (non-lung-cancer deaths) were examined microscopically in random order. There were 211 men who died in 1955-1960, of whom 154 smoked regularly, and 234 men who died in 1970-1977, of whom 181 were regular smokers. Changes studied included basal-cell hyperplasia, loss of cilia and occurrence of cells with atypical nuclei. In both periods studied these histologic changes occurred far less frequently in nonsmokers than in cigarette smokers and increased in frequency with amount of smoking, adjusted for age. Sections with advanced histologic changes in those dying in 1955-1960 occurred in 0 per cent of nonsmokers, in 2.6 per cent of those smoking one to 19 cigarettes a day, in 13.2 per cent of those smoking 20 to 39 and in 22.5 per cent of those smoking 40+ cigarettes a day. In those who died in 1970-1977 the percentages were 0, 0.1, 0.8, and 2.2, respectively.


A novel report of hatching plasticity in the phylum Echinodermata.
Hatching plasticity occurs in response to a wide range of stimuli across many animal taxa, including annelids, arthropods, mollusks, and chordates. Despite the prominence of echinoderms in developmental biology and more than 100 years of detailed examination of their development under a variety of conditions, environmentally cued hatching plasticity has never been reported in the phylum Echinodermata. Here we report plasticity in the timing and stage of hatching of embryos of the sand dollar Echinarachnius parma in response to reductions in salinity. Embryos of E. parma increased their time to hatching more than twofold in response to ecologically relevant salinity reductions, while maintaining an otherwise normal developmental schedule. Embryos that experienced the greatest delay in hatching time emerged from the fertilization envelope as four-arm pluteus larvae rather than hatching as blastulae or early gastrulae. Salinity manipulations across multiple male-female pairs indicated high variability in hatching time both within and among clutches, suggesting significant intraspecific variation in developmental responses to salinity.


Aetiology and risk factors related to traumatic dental injuries--a review of the literature.
During the past 30 years, the number of aetiologies of traumatic dental injuries (TDIs) has increased dramatically in the literature and now includes a broad spectrum of variables, including oral and environmental factors and human behaviour. The aim of this study is to present an international review of well-known as well as less well-known unintentional and intentional causes of TDIs. Moreover, some models that are useful in investigating contact sport injuries are presented. The databases of Medline, Cochrane, Social Citation Index, Science Citation Index and CINAHL from 1995 to the present were used. Oral factors (increased overjet with protrusion), environmental determinants (material deprivation) and human behaviour (risk-taking children, children being bullied, emotionally stressful conditions, obesity and attention-deficit hyperactivity disorder) were found to increase the risk for TDIs. Other factors increasing the risk for TDIs are presence of illness, learning difficulties, physical limitations and inappropriate use of teeth. A new cause of TDIs that is of particular interest is oral piercing. In traffic facial injury was similar in unrestrained occupants (no seat belts) and occupants restrained only with an air bag. Amateur athletes have been found to suffer from TDIs more often than professional athletes. Falls and collisions mask intentional TDIs, such as physical abuse, assaults and torture. Violence has increased in severity during the past few decades and its role has been underestimated when looking at intentional vs unintentional TDIs. There are useful models to prevent TDIs from occurring in sports. WHO Healthy Cities and WHO Health Promoting Schools Programmes offer a broad solution for dental trauma as a public health problem. The number of known causes of TDIs has grown to alarming levels, probably because of increased interest of the causes and the underlying complexity of a TDI. Accepted oral, environmental and human aetiological factors must therefore be included in the registration of TDIs.


[The use of cephalosporin antibiotics in the treatment and prevention of infections].
The general characteristics of cefalosporins was done, the principle of their division on generations, the pharmacokinetics differences of some preparations were indicated in the article. On the ground of microbiological data the perspective of klaforan, orelox, cefpirom application in the clinic was shown.


Thermally tunable ultracompact Fano resonator on a silicon photonic chip.
A thermally tunable ultracompact Fano resonator on a silicon photonic chip is reported. The Fano resonator is implemented by using an add-drop microdisk resonator (MDR) with the through and drop ports connected by two waveguides and combined via an adiabatic 2×2, 3 dB coupler to form a Mach-Zehnder interferometer (MZI). Due to the resonant mode interference between the MDR and the MZI, a Fano resonance with an asymmetric line shape resulted. A p-type-doped microheater is incorporated in the MDR to achieve thermal tunability. By tuning the direct current (DC) voltage applied to the microheater, the Fano resonance is tuned. The proposed Fano resonator is designed, fabricated, and characterized. Measurement results show that a Fano resonance with an extinction ratio of 30.2 dB and a slope rate of 41 dB/nm is achieved. When the microheater is tuned by tuning the DC voltage with a power from 0 to 22.9 mW, the Fano line shape is largely tuned with the Fano parameter q tuned from negative to positive and a maximum wavelength shifting as large as 15.97 nm. Thanks to its ultracompact configuration, and strong and fast tunability with low power consumption, the integrated Fano resonator holds a high potential for applications such as on-chip optical switching and sensing.


Systematic re-structure and new species of Sphaerodoridae (Annelida) after morphological revision and molecular phylogenetic analyses of the North East Atlantic fauna.
Detailed morphological study of more than 2600 North East Atlantic (NEA) sphaerodorids (SphaerodoridaeAnnelida) and phylogenetic analyses of DNA sequences of representatives of several identified morphospecies enforced changing the current systematic classification within the family allowed the discovery of new species provided new information about the morphological and genetic characterisation of members of this group and increased the species occurrence data to better infer their geographic and bathymetric distribution ranges. Phylogenetic analyses of nuclear (18S rRNA and 28S rRNA) and mitochondrial sequences (COI and 16S rRNA) of NEA short-bodied sphaerodorids revealed outstanding results including paraphyly of the genera Sphaerodoropsis Sphaerodoridium, and Sphaerephesia. The number of longitudinal and transverse rows of dorsal macrotubercles is proposed as potential synapomorphies for the main clades and are consequently herein used for the genera delimitation. The new classification proposed here implies nomenclatural changes and the erection of a new genus Geminofilum gen. n. to accommodate the species previously considered as Sphaerodoropsis with two transverse rows of dorsal macrotubercles per segment. Four species are being described herein: Euritmianordica Capa & Bakken sp. n. Sphaerephesiamultichaeta Capa Moreira & Parapar sp. n. Sphaerephesiaponsi Capa Parapar & Moreira sp. n. and Sphaerodoridiumceliae Moreira Capa & Parapar sp. n. Characterisation of the other 21 species including updated iconography and an identification key to all NEA short-bodied sphaerodorids are provided.


Structural characterization of the Mycobacterium tuberculosis biotin biosynthesis enzymes 7,8-diaminopelargonic acid synthase and dethiobiotin synthetase .
Mycobacterium tuberculosis (Mtb) depends on biotin synthesis for survival during infection. In the absence of biotin, disruption of the biotin biosynthesis pathway results in cell death rather than growth arrest, an unusual phenotype for an Mtb auxotroph. Humans lack the enzymes for biotin production, making the proteins of this essential Mtb pathway promising drug targets. To this end, we have determined the crystal structures of the second and third enzymes of the Mtb biotin biosynthetic pathway, 7,8-diaminopelargonic acid synthase (DAPAS) and dethiobiotin synthetase (DTBS), at respective resolutions of 2.2 and 1.85 A. Superimposition of the DAPAS structures bound either to the SAM analogue sinefungin or to 7-keto-8-aminopelargonic acid (KAPA) allowed us to map the putative binding site for the substrates and to propose a mechanism by which the enzyme accommodates their disparate structures. Comparison of the DTBS structures bound to the substrate 7,8-diaminopelargonic acid (DAPA) or to ADP and the product dethiobiotin (DTB) permitted derivation of an enzyme mechanism. There are significant differences between the Mtb enzymes and those of other organisms; the Bacillus subtilis DAPAS, presented here at a high resolution of 2.2 A, has active site variations and the Escherichia coli and Helicobacter pylori DTBS have alterations in their overall folds. We have begun to exploit the unique characteristics of the Mtb structures to design specific inhibitors against the biotin biosynthesis pathway in Mtb.


Primary structure of human complement component C2. Homology to two unrelated protein families.
The primary structure of the second component of human complement (C2) was determined by cDNA cloning and sequence analysis. C2 has 39% identity with the functionally analogous protein Factor B. The C-terminal half of C2a is homologous to the catalytic domains of other serine proteinases. C2b contains three direct repeats of approx. 60 amino acid residues. They are homologous to repeats in Factor B, C4b-binding protein and Factor H, suggesting a functional significance of the repeat in C4b and C3b binding. The repeats are also found in the non-complement proteins beta 2-glycoprotein I and interleukin-2 receptor, and this repeat family may be widespread.


Field trials with nitroscanate against cestodes and nematodes in dogs.
Clinical trials were carried out to test the safety and efficacy of the anthelmintic, nitroscanate, against naturally acquired helminth infections. The trials involved 418 dogs, of which 252 were treated with one or other of two different tablet formulations of nitroscanate. A coarse particle formulation given in two doses each of 200 mg per kg reduced the worm burden of Taenia spp and Dipylidium caninum by 100 per cent and Echinococcus granulosus by 94.1 per cent in farm dogs. Given as a single dose of 100 mg per kg this formulation reduced faecal worm egg counts by 95.3 per cent for Toxocara canis, 93.3 per cent for Toxascaris leonina and 99.1 per cent for Uncinaria stenocephala. A micronised formulation given in a single dose of 50 mg per kg caused a 97.9 per cent reduction in worm burden for Taenia spp and D caninum and 98.3 per cent reduction for E granulosus in foxhounds. The same formulation at this dosage gave reductions in faecal worm egg counts of 96.4 per cent for T canis and 100 per cent for T leonina and U stenocephala. Side-effects recorded were fewer than with piperazine and bunamidine.


Boron-containing coating yields enhanced antimicrobial and mechanical effects on translucent zirconia.
To evaluate the mechanical and antimicrobial properties of boron-containing coating on translucent zirconia (5Y-PSZ). 5Y-PSZ discs (Control) were coated with a glaze (Glaze), silver- (AgCoat), or boron-containing (BCoat) glasses. The coatings' antimicrobial potential was characterized using S. mutans biofilms after 48 h via viable colony-forming units (CFU), metabolic activity (CV) assays, and quantification of extracellular polysaccharide matrix (EPS). Biofilm architectures were imaged under scanning electron and confocal laser scanning microscopies (SEM and CLSM). The cytocompatibility was determined at 24 h via WST-1 and LIVE&DEAD assays using periodontal ligament stem cells (PDLSCs). The coatings' effects on properties were characterized by Vickers hardness, biaxial bending tests, and fractography analysis. Statistical analyses were performed via one-way ANOVA, Tukey's tests, Weibull analysis, and Pearson's correlation analysis. BCoat significantly decreased biofilm formation, having the lowest CFU and metabolic activity compared with the other groups. BCoat and AgCoat presented the lowest EPS, followed by Glaze and Control. SEM and CLSM images revealed that the biofilms on BCoat were thin and sparse, with lower biovolume. In contrast, the other groups yielded robust biofilms with higher biovolume. The cytocompatibility was similar in all groups. BCoat, AgCoat, and Glaze also presented similar hardness and were significantly lower than Control. BCoat had the highest flexural strength, characteristic strength and Weibull parameters (σF: 625 MPa; σ0: 620 MPa; m = 11.5), followed by AgCoat (σF: 464 MPa; σ0: 478 MPa; m = 5.3). BCoat is a cytocompatible coating with promising antimicrobial properties that can improve the mechanical properties and reliability of 5Y-PSZ.


An audit of a diverse community for safe routes to age in place: environmental policy implications.
Physical and cognitive limitations often accompany aging, increasing the importance of a safe and supportive environment to help older adults maintain mobility. Neighborhood design and maintenance must be evaluated to promote physical activity, mobility, and safety. Audit tools, geographic information system data, and resident interviews are used for this purpose, but often fail to provide information that can be translated to practice. The current project is part of a larger Miami-Dade Age-Friendly Initiative to create a metropolitan area that fosters a healthy environment for diverse adults of all ages and abilities. Safe Routes uses a toolkit based on the 5-E model providing practical resources to guide stakeholders in meeting the needs of the community. Findings include the Centers for Disease Control Healthy Aging Research Network Audit Tool assessment for environmental walkability factors. Results from street segment audits along with input from residents can be used to inform sound environmental policies.


Nitric oxide donor drugs: current status and future trends.
Nitric oxide synthesised in endothelial cells that line blood vessels has a wide range of functions that are vital for maintaining a healthy cardiovascular system. Reduced nitric oxide availability is implicated in the initiation and progression of many cardiovascular diseases and delivery of supplementary nitric oxide to help prevent disease progression is an attractive therapeutic option. Nitric oxide donor drugs represent a useful means of systemic nitric oxide delivery and organic nitrates have been used for many years as effective therapies for symptomatic relief from angina. However, nitrates have limitations and a number of alternative nitric oxide donor classes have emerged since the discovery that nitric oxide is a crucial biological mediator. This review focuses on novel advances and possible future directions in nitric oxide donor drug development.


[Role of CD69 in the pathogenesis of inflammation].
CD69 is a type II membrane protein expressed as a homodimer composed of heavily glycosylated subunits. CD69 is known as an early activation marker antigen of lymphocytes,and its selective expression in inflammatory infiltrates suggests that it plays a role in the pathogenesis of inflammatory diseases. In order to address the role of CD69 in the pathogenesis of arthritis and allergic airway inflammation, we established CD69-deficient mice. CD69-deficient mice displayed a markedly attenuated arthritic inflammatory response and airway inflammation. The administration of anti-CD69 antibody inhibited the induction of the antigen-induced airway inflammation and hyperresponsiveness. These results indicate that CD69 plays a crucial role in the pathogenesis of arthritis and allergic airway inflammation and that CD69 could be a possible therapeutic target for arthritis and asthma in human patients.


Socio-demographic factors associated with pet ownership amongst adolescents from a UK birth cohort.
In developed nations, pet ownership is common within families. Both physical and psychological health benefits may result from owning a pet during childhood and adolescence. However, it is difficult to determine whether these benefits are due to pet ownership directly or to factors linked to both pet ownership and health. Previous research found associations between a range of socio-demographic factors and pet ownership in seven-year-old children from a UK cohort. The current study extends this research to adolescence, considering that these factors may be important to consider in future Human-Animal Interaction (HAI) research across childhood. The Avon Longitudinal Study of Parents and Children (ALSPAC) collected pet ownership data prospectively via maternal reports from gestation up to age 10 years old and via self-report retrospectively at age 18 for ages 11 (n = 3063) to 18 years old (n = 3098) on cats, dogs, rabbits, rodents, birds, fish, tortoise/turtles and horses. The dataset also contains a wide range of potential confounders, including demographic and socio-economic variables. The ownership of all pet types peaked at age 11 (80%) and then decreased during adolescence, with the exclusion of cats which remained constant (around 30%), and dogs which increased through 11-18 years (26-37%). Logistic regression was used to build multivariable models for ownership of each pet type at age 13 years, and the factors identified in these models were compared to previously published data for 7 year-olds in the same cohort. There was some consistency with predictors reported at age 7. Generally sex, birth order, maternal age, maternal education, number of people in the household, house type, and concurrent ownership of other pets were associated with pet ownership at both 7 and 13 years (the direction of association varied according to pet type). Factors that were no longer associated with adolescent pet ownership included child ethnicity, paternal education, and parental social class. A number of socio-demographic factors are associated with pet ownership in childhood and adolescence and they differ according to the type of pet, and age of child. These factors are potential confounders that must be considered in future HAI studies.


[A hospital cancer registry. Breast cancer, 1977].
The clinical histories of the patients studied in the National Institute of Oncology in 1977 were reviewed in relation to breast cancer in women, that was found in 52.5% of them. 50% were comprised between 40 and 60 years of age, 68.3% were diagnosed within the first year after the presentation of clinical symptoms. 30.6% of the patients had a localized tumor, while all the others showed regional lymph node involvement and/or distant metastases. Malignancy was confirmed histologically in 90 per 100 of the cases; 41.5% were carcinomas, 27.9% corresponding to the infiltrating duct carcinoma type. Therapy could be applied to 80% of the patients with a curative purpose, while in 20% was only palliative. Including those women with previous treatments in other medical centers before coming to the National Institute of Oncology, the treatment applied was surgery in 90%, radiotherapy in 60% and chemotherapy in 20% of the cases. Five-year survival was 43% (28% clinically free of disease, and 15% with symptoms of disease).


Chemical lesion of visceral afferents causes transient overconsumption of unfamiliar high-fat diets in rats.
Because it is commonly assumed that the major role of visceral afferents in food intake control is to terminate meals by carrying negative-feedback signals to the brain, we hypothesized that overconsumption should occur in rats with chemically lesioned visceral afferents if they were presented with an unfamiliar diet. Adult male Sprague-Dawley (SD) rats were treated with multiple doses of capsaicin or vehicle as a control. Five weeks later, a series of 3-h feeding tests after 24-h deprivation was carried out, first with chow and then with either a solid (vegetable shortening) or liquid (Ensure) unfamiliar high-fat diet. Both groups consumed similar amounts of their powdered chow maintenance diet, but capsaicin-treated rats consumed at least 50% more of either high-fat diet than vehicle controls (P < 0.01) at the beginning of the first trial. During second and third trials with the now-familiar high-fat diet, intake was no longer significantly different between the two groups, suggesting rapid engagement of redundant control mechanisms. These results support a role of capsaicin-sensitive visceral afferents in providing negative feedback for early meal termination during the ingestion of unfamiliar diets.


Mutations in genes encoding the glycine cleavage system predispose to neural tube defects in mice and humans.
Neural tube defects (NTDs), including spina bifida and anencephaly, are common birth defects of the central nervous system. The complex multigenic causation of human NTDs, together with the large number of possible candidate genes, has hampered efforts to delineate their molecular basis. Function of folate one-carbon metabolism (FOCM) has been implicated as a key determinant of susceptibility to NTDs. The glycine cleavage system (GCS) is a multi-enzyme component of mitochondrial folate metabolism, and GCS-encoding genes therefore represent candidates for involvement in NTDs. To investigate this possibility, we sequenced the coding regions of the GCS genes: AMT, GCSH and GLDC in NTD patients and controls. Two unique non-synonymous changes were identified in the AMT gene that were absent from controls. We also identified a splice acceptor site mutation and five different non-synonymous variants in GLDC, which were found to significantly impair enzymatic activity and represent putative causative mutations. In order to functionally test the requirement for GCS activity in neural tube closure, we generated mice that lack GCS activity, through mutation of AMT. Homozygous Amt(-/-) mice developed NTDs at high frequency. Although these NTDs were not preventable by supplemental folic acid, there was a partial rescue by methionine. Overall, our findings suggest that loss-of-function mutations in GCS genes predispose to NTDs in mice and humans. These data highlight the importance of adequate function of mitochondrial folate metabolism in neural tube closure.


Cases from the aerospace medicine residents' teaching file. Case #47. Primary pulmonary coccidioidomycosis.
This case report on primary pulmonary coccidioidomycosis outlines a typical presentation in an individual at high risk and the pitfalls that can occur to practitioners unfamiliar with this disease. The patient was a 36-year-old black male who presented with an initial abnormal chest X-ray and who eventually was correctly diagnosed and improved on a 30-day course of ketoconazole. This patient showed no signs of dissemination.


X-ray diffraction study of feline leukemia virus fusion peptide and lipid polymorphism.
The structural effects of the fusion peptide of feline leukemia virus (FeLV) on the lipid polymorphism of N-methylated dioleoylphosphatidylethanolamine were studied using a temperature ramp with sequential X-ray diffraction. This peptide, the hydrophobic amino-terminus of p15E, has been proven to be fusogenic and to promote the formation of highly curved, intermediate structures on the lamellar liquid-crystal to inverse hexagonal phase transition pathway. The FeLV peptide produces marked effects on the thermotropic mesomorphic behaviour of MeDOPE, a phospholipid with an intermediate spontaneous radius of curvature. The peptide is shown to reduce the lamellar repeat distance of the membrane prior to the onset of an inverted cubic phase. This suggests that membrane thinning may play a role in peptide-induced membrane fusion and strengthens the link between the fusion pathway and inverted cubic phase formation. The results of this study are interpreted in relation to models of the membrane fusion mechanism.


The JAK2 46/1 haplotype in Budd-Chiari syndrome and portal vein thrombosis.
The germline JAK2 46/1 haplotype has been associated with the development of JAK2(V617F)-positive as well as JAK2(V617F)-negative myeloproliferative neoplasms (MPNs). In this study we examined the role of the 46/1 haplotype in the etiology and clinical presentation of patients with splanchnic vein thrombosis (SVT), in which MPNs are the most prominent underlying etiological factor. The single-nucleotide polymorphism rs12343867, which tags 46/1, was genotyped in 199 SVT patients. The 46/1 haplotype was overrepresented in JAK2(V617F)-positive SVT patients compared with controls (P < .01). Prevalence of the 46/1 haplotype in JAK2(V617F)-negative SVT patients did not differ from prevalence in the controls. However, JAK2(V617F)-negative SVT patients with a proven MPN also exhibited an increased frequency of the 46/1 haplotype (P = .06). Interestingly, 46/1 was associated with increased erythropoiesis in JAK2(V617F)-negative SVT patients. We conclude that the 46/1 haplotype is associated with the development of JAK2(V617F)-positive SVT. In addition, our findings in JAK2(V617F)-negative SVT patients indicate an important role for the 46/1 haplotype in the etiology and diagnosis of SVT-related MPNs, independent of JAK2(V617F), that requires further exploration.


Surgical palliation for pancreatic cancer. The UCLA experience.
We reviewed the records of 340 patients with a tissue diagnosis of pancreatic cancer treated at UCLA Medical Center between 1973 and 1988. Sixty-one patients underwent pancreatic resection (group I), 173 had some form of surgical palliation (group II), and 106 had neither (group III). The diagnosis was made 1 to 2 months more quickly in the last 8 years of the review than in the first 8 years, but the effect of early diagnosis on curability was negligible. Biliary obstruction was best treated by cholecystojejunostomy or choledochojejunostomy, which were equally effective. Anastomoses to the jejunum were safer and more effective than were those to the duodenum for the relief of biliary obstruction. Gastrojejunostomy should be performed prophylactically as well as therapeutically. It was effective and safe in both settings. Surgical palliation for pancreatic cancer was generally effective and was associated with an operative mortality rate of less than 10%. However morbidity was high, with significant complications occurring in one third of cases.


Hyperamylasaemia and/or hyperlipasaemia: incidence and underlying causes in hospitalized patients with non-pancreatic diseases.
There are no recommendations on how to proceed in patients with increased pancreatic enzyme activities but non-pancreatic diseases. The objective of this study was to investigate prospectively in a general medical hospital the incidence, causes and clinical impact of hyperamylasaemia and/or hyperlipasaemia in these patients. During a 4-month period in 2004, amylase and lipase activities were measured in all patients (n, 1765) admitted to the Department of Internal Medicine of the Municipal Clinic of Lüneburg, Germany. Patients with increased enzyme activities underwent abdominal ultrasonography and, if inconclusive, other imaging procedures. Amylase and/or lipase activities were increased in 140 (8%) patients with non-pancreatic diseases: amylase in 29 cases (21%) and 1 case (1%)<3 and>3 times the upper limit of normal, respectively, lipase in 120 cases (86%) and 6 cases (4%)<3 and>3 times, respectively. In 53 (38%) patients no further imaging studies could be performed. In the remaining 87 patients amylase was increased in 18 patients (21%) and lipase in 3 patients (3%). The pancreas was normal on imaging procedures in 77 (89%) patients and abnormal in 10 (11%) (chronic pancreatitis: 4, small cystic lesions: 6). The cystic lesions were mostly located in the pancreas head, probably corresponding to branch-duct intraductal papillary mucinous neoplasm. Increased pancreatic enzyme activities occur in 8% of hospitalized patients with non-pancreatic diseases, but pancreatic abnormalities will be found in only a few (11%), which are unlikely to need further investigation or therapy. Therefore, routine analysis of amylase or lipase activities in patients with non-pancreatic diseases is not indicated.


A soluble isoform of the rhesus monkey nonclassical MHC class I molecule Mamu-AG is expressed in the placenta and the testis.
The nonclassical MHC class I locus HLA-G is expressed primarily in the placenta, although other sites of expression have been noted in normal and pathological situations. In addition, soluble HLA-G isoforms have been detected in the serum of pregnant and nonpregnant women as well as men. The rhesus monkey placenta expresses a novel nonclassical MHC class I molecule Mamu-AG, which has features remarkably similar to those of HLA-G. We determined that the rhesus placenta expresses Mamu-AG mRNA (Mamu-AG5), retaining intron 4 as previously noted in HLA-G5. Immunostaining experiments with Ab 16G1 against the soluble HLA-G5 intron 4 peptide demonstrated that an immunoreactive protein(s) was present in the syncytiotrophoblasts of the chorionic villi of the rhesus placenta, within villous cytotrophoblasts, and occasionally within cells of the villous stroma. The Mamu-AG5 mRNA was readily detected in rhesus testis (although not in ejaculated sperm). Whereas an Ab against membrane-bound Mamu-AG stained few cells, primarily in the interstitium of the testis, there was consistent immunostaining for Mamu-AG5 in cells within the seminiferous tubules, which was corroborated by localization of Mamu-AG mRNA by in situ hybridization. While primary spermatocytes were negative, Sertoli cells, spermatocytes, and spermatids were consistently positive for 16G1 immunostaining. The specific recognition of the soluble Mamu-AG isoform was confirmed by Western blotting of Mamu-AG5 expressed in heterologous cells. The results demonstrate that a soluble nonclassical MHC class I molecule is expressed in the rhesus monkey placenta and testis, and confirm and extend the unique homology between HLA-G and the rhesus nonclassical molecule Mamu-AG.


Us and them: memory advantages in perceptually ambiguous groups.
Ingroup advantages and outgroup deficits in perception and memory are well-established in research on race, gender, and other ostensibly identifiable social categories. The present study extended this research to a social category that is not as perceptually apparent: male sexual orientation. Consistent with hypotheses, an interaction of participant sexual orientation and image sexual orientation revealed an ingroup enhancement and outgroup deficit for memory of faces that participants perceived-both accurately and inaccurately--as belonging to either their ingroup or outgroup in a subsequent task. Additionally, parallel effects were found for the accurate identification of sexual orientation--a finding consistent with previous literature. The present data highlight the importance of social categorization for subsequent memory and suggest that the underlying cognitive machinery responsible for the recognition of groups may be co-opted for other relevant social applications.