Surgical complications of endoscopic sphincterotomy.
Endoscopic sphincterotomy (papillotomy) was performed in 289 patients for choledocholithiasis (250, of whom 223 had undergone cholecystectomy previously), papillary stenosis or spasm (32) and ampullary neoplasm (7). The complications encountered in 39 patients were hemorrhage (15 patients), perforation (4), hemorrhage and perforation (1), cholangitis (5), pancreatitis (11), impaction (1) and others (2). Laparotomy was required in seven of these patients for hemorrhage (two), perforation (two), hemorrhage and perforation (one), pancreatitis (one) and impaction (one). Bleeding required duodenotomy with an extension of the sphincterotomy incision to control hemorrhage, and a formal sutured sphincteroplasty. Perforation occurred at the junction of the distal bile duct and duodenum and was managed by mobilization of the duodenum, with T-tube drainage through the perforation, and sutured closure. A pancreatic abscess following pancreatitis required surgical drainage. An impacted Dormia basket with entrapped stone in the bile duct required duodenotomy for its removal. There was a high risk of perforation in those patients who did not have choledocholithiasis or who had had a previous Billroth II gastrectomy. There were two deaths but the overall complication rate of 2.4% is considered low, because many of the patients were elderly or debilitated.


The contribution of community health systems to resilience: Case study of the response to the drought in Ethiopia.
Ethiopia's exposure to the El Niño drought (2015-2016) resulted in high malnutrition, internally displaced people, and epidemics of communicable diseases, all of which strained the health system. The drought was especially challenging for mothers and children. We aimed to identify salient factors that can improve health system resilience by exploring the successes and challenges experienced by a community-based health system during the drought response. We collected data via key informant interviews and focus group discussions to capture diverse perspectives across the health system (eg, international, national, district, facility, and community perspectives). Data were collected from communities in drought-affected regions of: 1) Somali, Sitti Zone, 2) Hawassa, Southern Nations, Nationalities, and Peoples' Region (SNNPR), and 3) Tigray, Eastern Zone. Data were analysed using a deductive-inductive approach using thematic content analysis applied to a conceptual framework. A total of 94 participants were included (71 from the communities and 23 from other levels). Key themes included the importance of: 1) organized community groups linked to the health system, 2) an effective community health workforce within strong health systems, 3) adaptable human resource structures and service delivery models, 4) training and preparedness, and 5) strong government leadership with decentralized decision making. The results of this study provide insights from across the health system into the successes and challenges of building resilience in community-based health systems in Ethiopia during the drought. As climate change exacerbates extreme weather events, further research is needed to understand the determinants of building resilience from a variety of shocks in multiple contexts, especially focusing on harnessing the power of communities as reservoirs of resilience.


Trans-Golgi network syntaxin 10 functions distinctly from syntaxins 6 and 16.
Syntaxin 10 is a soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) protein localized to the trans-Golgi network (TGN), where two other members of the syntaxin family, syntaxins 6 and 16, also reside. The role of syntaxin 10 in regulating TGN protein traffic is not yet defined. Syntaxin 10 co-localizes well with syntaxins 6 and 16 at the TGN in interphase cells, and appears to interact with both syntaxin 6 and 16 as evidenced by co-immunoprecipitation analyses. However, unlike syntaxin 6 and 16, neither syntaxin 10 antibodies nor its cytosolic domain inhibits endosome-TGN transport of shiga toxin. Syntaxin 16 knockdown with small interfering RNA (siRNA) affects the TGN localization of syntaxin 6 but not syntaxin 10, and clearly inhibits endosome-TGN transport. On the other hand, knockdown of syntaxin 10 expressions had no significant effect on the TGN localization of syntaxin 6 and 16, and did not inhibit endosome-TGN transport. Unlike syntaxin 16, syntaxin 10 does not interact specifically with Vps45, the Sec1/Munc18 (SM) family member at the TGN. On the other hand, syntaxin 10 reciprocally co-immunoprecipitated endosomal syntaxin 12/13, and knockdown of syntaxin 10 expressions affects the surface expression of transferrin receptor (TfR) and seems to induce the formation of an immobile TfR pool. Therefore, in spite of its co-localization and possible interaction with syntaxin 16, syntaxin 10 is not part of the syntaxin 16-based SNARE complex involved in endosome-TGN transport, and may have a hitherto unrecognized function in the TGN-endosome boundary.


Randomized controlled trial of mefenamic acid vs paracervical block for relief of pain for outpatient uterine curettage.
To compare the efficacy of mefenamic acid vs paracervical block for pain relief during and after fractional curettage. Between January 1 and July 31, 2002, the authors enrolled 87 patients with abnormal uterine bleeding, who requested fractional curettage at the Outpatient Gynecologic Clinic, Srinagarind Hospital, Khon Kaen University. A simple randomization procedure was used to distribute the patients into a control group comprising 44 patients given a paracervical block and a treatment group comprising 43 patients given mefenamic acid (500 mg) 2 hours before starting the procedure. Pain was scored using a visual analogue scale (VAS range, 0 to 10). The median pain scores of the treatment types during endocervical, endometrial, immediately after, and 30 minutes after, fractional curettage were 2.5 vs 3.0 (p = 0.42), 6.5 vs 7.5 (p = 0.19), 4.0 vs 3.5 (p = 0.20) and 1.5 vs 1.0 (p = 0.17), respectively. The rate of complications was 6.8% (3 in 44) in the paracervical lignocaine injection group. The efficacy of pain relief for fractional curettage using oral mefenamic acid (500 mg) two hours before the procedure was not statistically different from the paracervical block, but there were fewer side effects. Mefenamic acid should be considered an alternate pain relief during fractional curettage.


Conversion of psoriasis patients from the conventional formulation of cyclosporin A to a new microemulsion formulation: a randomized, open, multicentre assessment of safety and tolerability.
To assess the safety, tolerability and efficacy of a new cyclosporin A (CyA) microemulsion formulation, Sandimmun Neoral (Neoral), in patients with severe psoriasis that was stable on CyA administered as Sandimmun (SIM). In this 24-week, open, randomized, prospective, multicentre trial, 28 patients continued on the same dosage of SIM, while 30 converted to Neoral at 2.5 mg/kg/day or a dosage equivalent to their pre-conversion SIM dosage. During the study, dosages could be adjusted to maintain efficacy, because of adverse events or after disease stabilization. The maximum permitted dosage for either formulation was 5.0 mg/kg/day. Primary efficacy criteria were change in Psoriasis Area and Severity Index (PASI) from baseline and time to relapse. The dosage was increased to maintain efficacy in 22 patients (Neoral 13; SIM 9) and 20 dose reductions for safety were required (Neoral 14, SIM 6). In both groups, PASI scores remained stable throughout and relapses were primarily a result of dosage reduction after disease stabilization. No significant difference was found between groups in the proportion of patients remaining relapse-free. Adverse events were recorded in 20 patients receiving Neoral and 14 receiving SIM. Most drug-related events were of mild or moderate severity and reflected the known CyA side-effect profile. Dose titration guidelines ensured that mean blood pressure and serum creatinine concentrations remained stable in both groups. If the guidelines for CyA use are followed and the Neoral dosage does not exceed 5 mg/kg/day, conversion of stable patients with severe psoriasis from SIM to Neoral should present no clinically relevant safety or tolerability problems and efficacy of treatment is maintained.


Transcriptional, posttranscriptional, and posttranslational regulation of SHOOT MERISTEMLESS gene expression in Arabidopsis determines gene function in the shoot apex.
The activity of SHOOT MERISTEMLESS (STM) is required for the functioning of the shoot apical meristem (SAM). STM is expressed in the SAM but is down-regulated at the site of leaf initiation. STM is also required for the formation of compound leaves. However, how the activity of STM is regulated at the transcriptional, posttranscriptional, and posttranslational levels is poorly understood. We previously found two conserved noncoding sequences in the promoters of STM-like genes across angiosperms, the K-box and the RB-box. Here, we characterize the function of the RB-box in Arabidopsis (Arabidopsis thaliana). The RB-box, along with the K-box, regulates the expression of STM in leaf sinuses, which are areas on the leaf blade with meristematic potential. The RB-box also contributes to restrict STM expression to the SAM. We identified FAR1-RELATED SEQUENCES-RELATED FACTOR1 (FRF1) as a binding factor to the RB-box region. FRF1 is an uncharacterized member of a subfamily of four truncated proteins related to the FAR1-RELATED SEQUENCES factors. Internal deletion analysis of the STM promoter identified a region required to repress the expression of STM in hypocotyls. Expression of STM in leaf primordia under the control of the JAGGED promoter produced plants with partially undifferentiated leaves. We further found that the ELK domain has a role in the posttranslational regulation of STM by affecting the nuclear localization of STM.


Nanomedicine-mediated cancer stem cell therapy.
Circumstantial evidence suggests that most tumours are heterogeneous and contain a small population of cancer stem cells (CSCs) that exhibit distinctive self-renewal, proliferation and differentiation capabilities, which are believed to play a crucial role in tumour progression, drug resistance, recurrence and metastasis in multiple malignancies. Given that the existence of CSCs is a primary obstacle to cancer therapy, a tremendous amount of effort has been put into the development of anti-CSC strategies, and several potential approaches to kill therapeutically-resistant CSCs have been explored, including inhibiting ATP-binding cassette transporters, blocking essential signalling pathways involved in self-renewal and survival of CSCs, targeting CSCs surface markers and destroying the tumour microenvironment. Meanwhile, an increasing number of therapeutic agents (e.g. small molecule drugs, nucleic acids and antibodies) to selectively target CSCs have been screened or proposed in recent years. Drug delivery technology-based approaches hold great potential for tackling the limitations impeding clinical applications of CSC-specific agents, such as poor water solubility, short circulation time and inconsistent stability. Properly designed nanocarrier-based therapeutic agents (or nanomedicines) offer new possibilities of penetrating CSC niches and significantly increasing therapeutic drug accumulation in CSCs, which are difficult for free drug counterparts. In addition, intelligent nanomedicine holds great promise to overcome pump-mediated multidrug resistance which is driven by ATP and to decrease detrimental effects on normal somatic stem cells. In this review, we summarise the distinctive biological processes related to CSCs to highlight strategies against inherently drug-resistant CSCs. We then focus on some representative examples that give a glimpse into state-of-the-art nanomedicine approaches developed for CSCs elimination. A perspective on innovative therapeutic strategies and the potential direction of nanomedicine-based CSC therapy in the near future is also presented.


Solute Partitioning in Micelles: Combining Molecular Dynamics Simulations, COSMOmic, and Experiments.
The partition equilibria of solutes between micelles and an aqueous phase is a key factor in many applications. Depending on the task, many micelle-solute combinations are possible. Therefore, theoretical methods to predict the partition behavior in micellar systems are needed. Here, two predictive methods are evaluated and compared. First, it is shown how molecular dynamics simulations (MD) with the umbrella sampling method can be used to calculate free energy profiles in micellar systems. The second applied method is an extension of the COSMO-RS theory to anisotropic systems termed COSMOmic. Both methods are compared by means of free energy profiles and experimental micelle/water partition coefficients. A particular focus is on the partitioning of ionized solutes. As experimental data for partitioning in micelles especially for charged solutes is rare, partition coefficients were also determined experimentally. To get a general understanding of micelles examples of all micelle classes (classified by headgroup charge) are studied: nonionic Triton X-114 (TX114), zwitterionic miltefosine (HePC), anionic sodium dodecyl sulfate (SDS), and cationic cetyltrimethylammonium bromid (CTAB). The free energy profiles of neutral solutes obtained from MD simulations and COSMOmic are in an overall good agreement, and partition coefficients from both methods are in good agreement with experimental data. Depending on the system, the results for charged solutes show some deviations between the methods and experimental data.


Maximum-entropy closure of hydrodynamic moment hierarchies including correlations.
Generalized hydrodynamic moment hierarchies are derived which explicitly include nonequilibrium two-particle and higher-order correlations. The approach is adapted to strongly correlated media and nonequilibrium processes on short time scales which necessitate an explicit treatment of time-evolving correlations. Closure conditions for the extended moment hierarchies are formulated by a maximum-entropy approach, generalizing related closure procedures for kinetic equations. A self-consistent set of nonperturbative dynamical equations are thus obtained for a chosen set of single-particle and two-particle (and possibly higher-order) moments. Analytical results are derived for generalized Gaussian closures including the dynamic pair distribution function and a two-particle correction to the current density. The maximum-entropy closure conditions are found to involve the Kirkwood superposition approximation.


Impact of Paracoccin Gene Silencing on Paracoccidioides brasiliensis Virulence.
Among the endemic deep mycoses in Latin America, paracoccidioidomycosis (PCM), caused by thermodimorphic fungi of the Paracoccidioides genus, is a major cause of morbidity. Disease development and its manifestations are associated with both host and fungal factors. Concerning the latter, several recent studies have employed the methodology of gene modulation in P. brasiliensis using antisense RNA (AsRNA) and Agrobacterium tumefaciens-mediated transformation (ATMT) to identify proteins that influence fungus virulence. Our previous observations suggested that paracoccin (PCN), a multidomain fungal protein with both lectin and enzymatic activities, may be a potential P. brasiliensis virulence factor. To explore this, we used AsRNA and ATMT methodology to obtain three independent PCN-silenced P. brasiliensis yeast strains (AsPCN1, AsPCN2, and AsPCN3) and characterized them with regard to P. brasiliensis biology and pathogenicity. AsPCN1, AsPCN2, and AsPCN3 showed relative PCN expression levels that were 60%, 40%, and 60% of that of the wild-type (WT) strain, respectively. PCN silencing led to the aggregation of fungal cells, blocked the morphological yeast-to-mycelium transition, and rendered the yeast less resistant to macrophage fungicidal activity. In addition, mice infected with AsPCN1, AsPCN2, and AsPCN3 showed a reduction in fungal burden of approximately 96% compared with those inoculated with the WT strain, which displayed a more extensive destruction of lung tissue. Finally, mice infected with the PCN-silenced yeast strains had lower mortality than those infected with the WT strain. These data demonstrate that PCN acts as a P. brasiliensis contributory virulence factor directly affecting fungal pathogenesis.IMPORTANCE The nonexistence of efficient genetic transformation systems has hampered studies in the dimorphic fungus Paracoccidioides brasiliensis, the etiological agent of the most frequent systemic mycosis in Latin America. The recent development of a method for gene expression knockdown by antisense RNA technology, associated with an Agrobacterium tumefaciens-mediated transformation system, provides new strategies for studying P. brasiliensis Through this technology, we generated yeasts that were silenced for paracoccin (PCN), a P. brasiliensis component that has lectin and enzymatic properties. By comparing the phenotypes of PCN-silenced and wild-type strains of P. brasiliensis, we identified PCN as a virulence factor whose absence renders the yeasts unable to undergo the transition to mycelium and causes a milder pulmonary disease in mice, with a lower mortality rate. Our report highlights the importance of the technology used for P. brasiliensis transformation and demonstrates that paracoccin is a virulence factor acting on fungal biology and pathogenesis.


Correction of confidence intervals in excess relative risk models using Monte Carlo dosimetry systems with shared errors.
In epidemiological studies, exposures of interest are often measured with uncertainties, which may be independent or correlated. Independent errors can often be characterized relatively easily while correlated measurement errors have shared and hierarchical components that complicate the description of their structure. For some important studies, Monte Carlo dosimetry systems that provide multiple realizations of exposure estimates have been used to represent such complex error structures. While the effects of independent measurement errors on parameter estimation and methods to correct these effects have been studied comprehensively in the epidemiological literature, the literature on the effects of correlated errors, and associated correction methods is much more sparse. In this paper, we implement a novel method that calculates corrected confidence intervals based on the approximate asymptotic distribution of parameter estimates in linear excess relative risk (ERR) models. These models are widely used in survival analysis, particularly in radiation epidemiology. Specifically, for the dose effect estimate of interest (increase in relative risk per unit dose), a mixture distribution consisting of a normal and a lognormal component is applied. This choice of asymptotic approximation guarantees that corrected confidence intervals will always be bounded, a result which does not hold under a normal approximation. A simulation study was conducted to evaluate the proposed method in survival analysis using a realistic ERR model. We used both simulated Monte Carlo dosimetry systems (MCDS) and actual dose histories from the Mayak Worker Dosimetry System 2013, a MCDS for plutonium exposures in the Mayak Worker Cohort. Results show our proposed methods provide much improved coverage probabilities for the dose effect parameter, and noticeable improvements for other model parameters.


Mechanism and Kinetics of Aztreonam Hydrolysis Catalyzed by Class-C β-Lactamase: A Temperature-Accelerated Sliced Sampling Study.
Enhanced sampling of large number of collective variables (CVs) is inevitable in molecular dynamics (MD) simulations of complex chemical processes such as enzymatic reactions. Because of the computational overhead of hybrid quantum mechanical/molecular mechanical (QM/MM)-based MD simulations, especially together with density functional theory, predictions of reaction mechanism, and estimation of free-energy barriers have to be carried out within few tens of picoseconds. We show here that the recently developed temperature-accelerated sliced sampling method allows one to sample large number of CVs, thereby enabling us to obtain rapid convergence in free-energy estimates in QM/MM MD simulation of enzymatic reactions. Moreover, the method is shown to be efficient in exploring flat and broad free-energy basins that commonly occur in enzymatic reactions. We demonstrate this by studying deacylation and reverse acylation reactions of aztreonam drug catalyzed by a class-C β lactamase (CBL) bacterial enzyme. Mechanistic details and nature of kinetics of aztreonam hydrolysis by CBL are elaborated here. The results of this study point to characteristics of the aztreonam drug that are responsible for its slow hydrolysis.


The growth and aflatoxin production of Aspergillus flavus strains on a cheese model system are influenced by physicochemical factors.
Traditional cheeses may be contaminated by aflatoxin-producing Aspergillus flavus during the ripening process, which has not been sufficiently taken into account. The objectives of this study were to evaluate the influence of water activity (aw), pH, and temperature on the lag phases, growth, and aflatoxin production of 3 A. flavus strains (CQ7, CQ8, and CG103) on a cheese-based medium. The results showed that the behavior of A. flavus strains was influenced by pH, aw, and temperature conditions. The CQ7 strain showed the maximum growth at pH 5.5, 0.99 aw, and 25°C, whereas for CQ8 and CQ103 strains, no differences were obtained at pH 5.5 and 6.0. In general, low pH, aw, and temperature values increased the latency times and decreased the growth rate and colony diameter, although aw and temperature were the most limiting factors. Maximum aflatoxin production on the cheese-based medium occurred at pH 5.0, 0.95 aw, and 25 or 30°C, depending on the strain. This study shows the effect of pH, aw, and temperature factors on growth and aflatoxin production of 3 aflatoxigenic A. flavus strains on a cheese-based medium. The findings may help to design control strategies during the cheesemaking process and storage, to prevent and avoid aflatoxin contamination by aflatoxigenic molds.


Differential effect of cadmium and mercury on growth and metabolism of Solanum melongena L. seedlings.
The present investigation reports the results of the effect of cadmium and mercury individually on seed germination, seedling growth, number of lateral roots, fresh and dry weights and seedling metabolism in Solanum melongena. Effect of different concentrations of these two heavy metals (Cadmium--50, 100, 300, 500, 700, 1000, 3000, 5000, 7000 and 9000 ppm and Mercury--5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 ppm) and durations for 6, 12 and 24 h were employed for all seedling parameters of brinjal. Both Cd and Hg showed drastic effects at high concentrations and longer duration with regard to seedling growth and metabolism.


[Influence of arterial hypertension on diabetic retinopathy].
The influence of arterial hypertension on the prevalence of diabetic retinopathy was evaluated by a cross-sectional study in 882 diabetic patients of whom 337 were insulin-treated and 505 were non insulin-treated. Arterial hypertension was defined by blood pressure values higher than 160-90 mmHg. Retinopathy was considered to be present when at least 2 microaneurysms were observed at the posterior pole. When duration of the diabetes was taken into account the prevalence of retinopathy in hypertensive subjects (69%) was not significantly higher than in normotensive subjects (47%) among the insulin-treated patients. However among non insulin-treated patients retinopathy was significantly more frequent in hypertensive (39%), than in normotensive subjects (25%; p less than 0.05).


Genomic analysis reveals complex population structure within the smooth newt, Lissotriton vulgaris, in Central Europe.
Species with wide-range distributions usually display high genetic variation. This variation can be partly explained by historical lineages that were temporally isolated from each other and are back into secondary reproductive contact, and partly by local adaptations. The smooth newt (Lissotriton vulgaris) is one of the most widely distributed amphibians species across Eurasia and forms a species complex with a partially overlapping distribution and morphology. In the present study, we explored the population genomic structure of smooth newt lineages in the Carpathian Basin (CB) relying on single-nucleotide polymorphisms. Our dataset included new and previously published data to study the secondary contact zone between lineages in the CB and also tested for the barrier effect of rivers to gene flow between these lineages. We confirmed the presence of the South L. v. vulgaris Lineage distributed in Transdanubia and we provided new distribution records of L. v. ampelensis inhabiting the eastern territories of the CB. High genetic diversity of smooth newts was observed, especially in the North Hungarian Mountains and at the interfluves of the main rivers in the South with four distinct lineages of L. v. vulgaris and one lineage of L. v. ampelensis showing a low level of admixture with the spatially closest L. v. vulgaris lineage. Moreover, admixture detected at the interfluve of the main rivers (i.e. Danube and Tisza) suggested a secondary contact zone in the area. Finally, we found that the river Danube has a very weak effect on population divergence, while the river Tisza is a geographical barrier limiting gene flow between smooth newt lineages. As the range boundaries of L. v. vulgaris and L. v. ampelensis in the CB coincide with the river Tisza, our study underpins the influence of rivers in lineage diversification.


Pancreatic cancer: medical aspects.
Although pancreatic cancer (PC) continues to be a formidable disease, numerous treatment strategies are evolving that we hope will result in improved patient survival. To prolong the survival of patients with PC, it is essential to detect PC at the earliest stage possible and to develop effective nonsurgical treatments for this disease. The current strategies for the early diagnosis of PC include the development of diagnostic modalities and screening programs for the early detection of PC and the determination of high-risk groups for PC. K-ras mutations in pancreatic juice obtained endoscopically have been studied recently in association with the early diagnosis of PC, although the interpretation of the presence of a mutated K-ras gene requires caution. The role of the current nonsurgical treatments for PC has been limited. Identification of an effective new chemotherapeutic agent is a high priority, and the enrollment of patients with PC with metastatic disease into well-designed clinical trials is essential. New targets for therapy based on the understanding of the molecular biology of PC may provide avenues for future trials. We must continue to search actively for more accurate methods of diagnosis and more effective methods of treatment for PC.


Three-dimensional ultrasound does not improve diagnosis of retained placental tissue compared to two-dimensional ultrasound.
The study objective was to improve ultrasonic diagnosis of retained placental tissue by measuring the volume of the uterine body and cavity using three-dimensional (3D) ultrasound. Twenty-five women who were to undergo surgical curettage due to suspected retained placental tissue were included. The volume of the uterine body and cavity was measured using the VOCAL imaging program. Twenty-one women had retained placental tissue histologically verified. Three of these had uterine volumes exceeding the largest volume observed in the normal puerperium. Seventeen of the 21 women had a uterine cavity volume exceeding the largest volume observed in the normal puerperium. In all 14 cases examined 28 days or more after delivery the cavity volume exceeded the largest volume observed in the normal puerperium. A large cavity volume estimated with 3D ultrasound is indicative of retained placental tissue. However, 3D ultrasound adds little or no diagnostic power compared to 2D ultrasound.


[A social psychiatric study on foreign-born inpatients at Tokyo Metropolitan Matsuzawa hospital].
The authors carried out a chart review study of 293 foreign-born patients who were admitted to Tokyo Metropolitan Matsuzawa Hospital from January 1986 to December 1992. We divided the subjects into three groups according to their living status in Japan, Immigrant group: 66 long-term residents, Resident group: 188 short-term residents with purposes of work or study, and Visitor group: 39 temporary visitors for sightseeing or business. The subjects came from 43 countries in the world, 70.3% (206/293) of whom were those from Asian countries. The subjects grew in number every year and the noticeable feature was a drastic increase in the number of Resident group (e.g. 6 patients in 1986, 63 in 1992). The mean duration (SD) of hospitalization for Immigrant group was the longest among the groups (Immigrant group: 58.7 (55.0) days, Resident group: 31.9 (35.0) days, Visitor group: 19.0 (16.3) days). The frequent diagnoses at discharge based on ICD-9, were schizophrenia (59.7%) and reactive psychosis (19.1%). The significantly larger proportion of Visitor group had previous psychiatric histories in their homeland than other groups (Visitor group: 56.4%, Immigrant group: 24.2%, Resident group: 23.9%). 70.0% (205/293) of the subjects could not use Japanese in usual settings and the number of them had been increasing every year. This communication difficulty had caused serious inconvenience in the treatment. All of Visitor group, and 71.8% (135/188) of Resident group could not receive social securities, and there were difficulties for them to raise money for their medical treatment and living expenses. 92.3% among Visitor group, 91.0% of Resident group and 21.2% of Immigrant group returned to their homeland after inpatient treatment. The result indicated that we must immediately establish the interpreter dispatch system for foreign-born patients and that of the social securities guaranteeing their medical care and living expenses.


Electrochemical CO2 reduction with low overpotential by a poly(4-vinylpyridine) electrode for application to artificial photosynthesis.
Pyridine molecules have been used as a catalyst to reduce the activation energy of the CO2 reduction reaction. It has been reported that CO2 is reduced by pyridine catalysts at low overpotential around -0.58 V vs. SCE. Poly(4-vinylpyridine), which has pyridine functional groups shows similar catalytic properties to reduce CO2 at low overpotential like pyridinium catalysts. Different thickness of P(4-VP) coated Pt electrodes were analyzed to determine the catalytic properties for CO2 reduction. Cyclic voltammetry, chronoamperometry and electrochemical impedance spectroscopy methods showed the catalytic CO2 reduction properties of a P(4-VP)/Pt electrode. Thin P(4-VP)/Pt film showed a low current density of -0.16 mA cm-2 under CO2 atmosphere and the current density reached -0.45 mA cm-2 with increase of the P(4-VP) thickness. The increase of current density was explained by an increased surface concentration of adsorbed pyridinium groups of the thick P(4-VP) layer. Nyquist plots also showed decrease of impedance with increase of the P(4-VP) layer indicating fast charge transfer between Pt and the P(4-VP) layer due to the increase of hybrid ionic complex formation on the Pt surface. However, charge transfer is restricted when the P(4-VP) layer becomes more thick because of slowed protonation of pyridine groups adjacent to the Pt surface due to the suppressed permeability of electrolyte solution into the PVP membrane. This electrochemical observation provides a new aspect of P(4-VP) polymer for CO2 reduction.


[Bioactivity and heterogeneity of human growth hormone in urine from acromegalic patients].
Bioactivity of hGH in urine from five acromegalic patients was determined by Nb2 rat lymphoma bioassay (Nb2BA) and IM-9 receptor modulation assay (RMA). One urine sample (case 1) was concentrated by dialysis and lyophilization. Bioactivity by IM-9 RMA showed close correlations with immunoactivity by RIA in serial dilutions (RMA/RIA; 0.66-1.77). The estimates of bioactivity by Nb2BA were higher in all diluted samples except undiluted one than those of immunoactivity. Four samples were concentrated by affinity chromatography, dialysis and lyophilization. The ratio of RMA and RIA estimates was between 0.81-1.24, and that of Nb2BA and RIA estimates was between 0.46-2.62. Heterogeneity of urinary hGH was analyzed by gelfiltration using TSK-G-3000SW column by high performance liquid chromatography (HPLC) and by sensitive sandwich enzyme immunoassay. In addition to the main peak of 22K molecular weight (more than 97%), very small peaks of 40-60K (big hGH) and 150K (big-big hGH) were detected in chromatographic profile. Urinary hGH from acromegalic patients was bioactive in IM-9 RMA and Nb2BA. The predominant form of urinary hGH from acromegalic patients was little hGH, but big and big-big hGH were detected by sensitive sandwich enzyme immunoassay.


Solid breast masses diagnosed as fibroadenoma at fine-needle aspiration biopsy: acceptable rates of growth at long-term follow-up.
To determine what growth rate is acceptable before recommending histologic diagnosis of solid breast lesions diagnosed as fibroadenoma at fine-needle aspiration biopsy (FNAB). For 1,070 consecutive patients with breast lesions diagnosed as fibroadenoma at FNAB, three measurements of each mass were performed at the initial visit when FNAB was performed and at each follow-up ultrasonographic examination. Changes in volumes were calculated. At one or more visits, 194 masses showed an increase in volume. Nonfibroadenomas were excluded, and the data were used for comparison. Percentiles (90th and 95th) for percentage change in volume per month were used to determine acceptable changes in dimensions (specifically, greatest anteroposterior, parallel-to-skin, and perpendicular-to-skin dimensions). There were 567 interval measurements of 179 masses in 173 patients younger than 50 years and 50 measurements of 15 masses in 14 patients 50 years or older at the time of FNAB. The 95th percentile for percentage change in volume per month was approximately 16% for patients younger than 50 years; the 90th percentile was approximately 13% for patients 50 years or older. The 95th percentile mean change in dimension in a 6-month interval for those younger than 50 years was 20%; the 90th percentile change for those 50 years or older was also 20%. All excised masses with slower growth proved benign at histologic examination. Solid breast masses diagnosed as fibroadenomas at FNAB may be safely followed up if volume growth rate is less than 16% per month in those younger than 50 years and less than 13% per month in those 50 years or older. Acceptable mean change in dimension for a 6-month interval is 20% for all ages.


Negative first impression judgements of autistic children by non-autistic adults.
Although autism inclusion and acceptance has increased in recent years, autistic people continue to face stigmatization, exclusion, and victimization. Based on brief 10-second videos, non-autistic adults rate autistic adults less favourably than they rate non-autistic adults in terms of traits and behavioural intentions. In the current study, we extended this paradigm to investigate the first impressions of autistic and non-autistic children by non-autistic adult raters and examined the relationship between the rater's own characteristics and bias against autistic children. Segments of video recorded interviews from 15 autistic and 15 non-autistic children were shown to 346 undergraduate students in audio with video, audio only, video only, transcript, or still image conditions. Participants rated each child on a series of traits and behavioural intentions toward the child, and then completed a series of questionnaires measuring their own social competence, autistic traits, quantity and quality of past experiences with autistic people, and explicit autism stigma. Overall, autistic children were rated more negatively than non-autistic children, particularly in conditions containing audio. Raters with higher social competence and explicit autism stigma rated autistic children more negatively, whereas raters with more autistic traits and more positive past experiences with autistic people rated autistic children more positively. These rapid negative judgments may contribute to the social exclusion experienced by autistic children. The findings indicate that certain personal characteristics may be related to more stigmatised views of autism and decreased willingness to interact with the autistic person. The implications of the findings are discussed in relation to the social inclusion and well-being of autistic people.


Hierarchy of metabolic compounds based on their synthesising capacity.
The concept of scopes is applied to analyse large metabolic networks. Scopes are defined as sets of metabolites that can be synthesised by a metabolic network when it is provided with given seeds (Sets of initial metabolic compounds). Thus, scopes represent synthesising capacities of the seeds in the network. A hierarchy is discussed in the sense that compounds, which are part of the scope of another compound, possess scopes themselves that are subsets of the former scope. This hierarchy is analysed by means of a directed acyclic graph. Using a simple chemical model, it is found that this hierarchy contains specific structures that can, to a large extent, be explained by the chemical composition of the participating compounds. In this way, it represents a new kind of map of metabolic networks, arranging the metabolic compounds according to their chemical capacity.


Direct estimation of kinetic parametric images for dynamic PET.
Dynamic positron emission tomography (PET) can monitor spatiotemporal distribution of radiotracer in vivo. The spatiotemporal information can be used to estimate parametric images of radiotracer kinetics that are of physiological and biochemical interests. Direct estimation of parametric images from raw projection data allows accurate noise modeling and has been shown to offer better image quality than conventional indirect methods, which reconstruct a sequence of PET images first and then perform tracer kinetic modeling pixel-by-pixel. Direct reconstruction of parametric images has gained increasing interests with the advances in computing hardware. Many direct reconstruction algorithms have been developed for different kinetic models. In this paper we review the recent progress in the development of direct reconstruction algorithms for parametric image estimation. Algorithms for linear and nonlinear kinetic models are described and their properties are discussed.


[Pleurodesis in malignant pleural effusion].
About 50% of the pleural effusions diagnosed are caused by a malignancy, especially by thoracic, pulmonary and ovarian cancer and lymphomas. The accumulation of fluid is caused by metastasization to the pleura and obstruction of lymph vessels and nodes. The effusion generally decreases if the tumour responds to systemic treatment. However, frequently this does not occur and the fluid has to be removed, to alleviate symptoms such as dyspnoea, coughing and a heavy sensation in the chest. Possible surgical therapies are draining through a needle or a drain, (partial) pleural resection and the creation of a pleuro-peritoneal shunt. Disadvantages of these are early recurrences, the severity of the intervention and (or) the high morbidity and mortality. The current standard treatment is pleurodesis brought about by a sclerosing agent, usually via a drain. The substances preferably used for this purpose are, in the order of decreasing importance, tetracycline, bleomycin or talc, doxycycline or minocycline. The most frequent adverse effects are chest pain and fever during and after the pleurodesis.


Stomatherapy and the stomatherapist.
Stomatherapy can be defined as specialist care given to patients who undergo surgery with construction of a colostomy, ileostomy or urinary conduit. The role of the stomatherapist is outlined and the stomatherapy techniques and appliances used by the Groote Schuur Hospital Stomatherapy Service are described.


Albuminuria: Prevalence, associated risk factors and relationship with cardiovascular disease.
To investigate the prevalence and associated risk factors of microalbuminuria, and to explore the relationship between albuminuria and cardiovascular disease (CVD). A nationally representative sample of 38,203 Chinese participants was categorized by different levels of urinary albumin-to-creatinine ratio (ACR; 0 -10 mg/g, 10 -20 mg/g, 20 -30 mg/g, 30 -300 mg/g). The prevalence of albuminuria was compared by using a single urinary ACR cut-off point and by sex-specific ACR cut-off points. Factors associated with the presence of albuminuria, and the relationship between albuminuria and CVD were analyzed by logistic regression. Prevalence of albuminuria as measured by a single ACR cut-point was significantly lower for men compared with women (13.9% vs 19.1% in the normal glucose tolerance group; 20.8% vs 26.8% in the impaired glucose tolerance group, P < 0.01). The prevalence of albuminuria, as measured by sex-specific ACR cut-points, was higher for men than women (31.4% vs 29.6% in the normal glucose tolerance group; 42.2% vs 39.3% in the impaired glucose tolerance group, P < 0.01). The independent risk factors for the presence of albuminuria were aging, female sex, hypertension, hyperglycemia, obesity, dyslipidemia, insulin resistance and metabolic syndrome. The subdivided normal ACR group did not show a linear or statistically significant relationship with CVD after adjusting for conventional CVD risk factors (P > 0.05). The prevalence of albuminuria was high in the general Chinese population. Aging, female sex, hypertension, hyperglycemia, dyslipidemia, insulin resistance, obesity and metabolic syndrome were all independent risk factors for albuminuria. The causal relationship between ACR and CVD might require further follow-up investigation.


Inhibition of mitochondrial protein synthesis during early stages of aflatoxin B1-induced hepatocarcinogenesis.
Experiments were designed to determine the in vivo effects of a single 6-mg/kg dose aflatoxin B1 on rat liver mitochondrial transcription and translation processes. With the use of intact hepatocytes and also a highly active mitoplast system for incorporation, it was observed that both mitochondrial transcription and translation activities are inhibited progressively even after 24 hr of carcinogen administration. Electrophoretic patterns of mitochondrial translation products show some qualitative changes during early periods of carcinogen administration. At later stages (greater than 12 hr), however, there is a general inhibition of many of the products, although by this time there is a qualitative and quantitative recovery in the synthesis of mitochondrial proteins imported from the cytoplasm. These results, along with the data showing considerably high levels of aflatoxin B1 binding to mitochondrial DNA suggest that mitochondrial genetic system is one of the direct targets during experimental carcinogenesis.


Does preoperative positron emission tomography with computed tomography predict nodal status in endometrial cancer? A pilot study.
Fewer than 20% of women with endometrial cancer have positive nodes, and an accurate noninvasive imaging modality to assess lymph node status would be helpful in selecting those who need lymphadenectomy. The objective of this pilot study was to evaluate positron emission tomography with computed tomography (pet-ct) in predicting nodal status before surgery for endometrial cancer. Twelve patients were enrolled at a single tertiary care centre. The sensitivity and specificity of preoperative pet-ct in predicting nodal status were 53.3% and 99.6% respectively. Using pet-ct, all metastatic nodes may not necessarily be detected, especially nodes with microscopic disease. The sensitivity of this imaging modality has to be improved before it can routinely be used in the preoperative evaluation of endometrial cancer.


Optic nerve head drusen. High-resolution computed tomographic approach.
Optic nerve head drusen are rare, inherited concretions, which are almost always calcified; their appearance must be known because they represent one of the major causes of pseudopapilledema. The computed tomographic (CT) scan can show small and buried drusen, which are sometimes difficult to diagnose by the ophthalmoscopic examination. Four cases of drusen (two bilateral and two unilateral) were confirmed or diagnosed by high-resolution CT. The CT appearance of drusen is characteristic because the calcifications are well defined, punctate, and strictly located in the optic disc. The use of high-resolution CT scanners is very helpful.


Comparison of intracorporeal knot-tying suture (polyglactin) and titanium endoclips in laparoscopic appendiceal stump closure: a prospective randomized study.
Laparoscopic appendectomy is a well-described surgical technique. However, concerns still exist regarding whether the closure of the appendiceal stump should be done with a clip, an endoloop, staples, or other techniques. Therefore, several modifications to the original technique with new materials have been introduced for appendiceal stump closure. The aim of this study was to compare intracorporeal (polyglactin) knot-tying suture with titanium endoclips in appendiceal stump closure during laparoscopic appendectomy. The study was carried out as a prospective randomized clinical trial between April 2010 and February 2011. Patients with a presentation of appendicitis were included into the study. Two groups were defined-patients with the titanium endoclip and patients with the knot-tying (polyglactin) suture. The results in terms of operating time, complication rates, and hospital stay were analyzed. Sixty-one patients who underwent laparoscopic appendectomy were enrolled in the titanium endoclip group (n=30) or the knot-tying (polyglactin) suture group (n=31). No statistically significant differences were detected between the groups in terms of the distribution of age, sex percentage, appendix localization, and histopathologic diagnosis (P>0.05). One patient required a second operation on postoperative day 10 because of intraperitoneal abscess. The mean operative time for the endoclip group (41.27 ± 12.2 min) was shorter than that for the knot-tying group (62.81 ± 15.4 min) (P=0.001). No statistically significant differences were detected between the groups in terms of hospital stay, follow-up time, and preoperative and postoperative complications (P>0.05). In laparoscopic appendectomy, using a titanium endoclip for optimizing and controlling the appendiceal stump closure is safe and is associated with shorter operation time. This also simplifies the procedure, so it can be a useful alternative to intracorporeal knot-tying for appendiceal stump closure.


Parametric functional principal component analysis.
Functional principal component analysis (FPCA) is a popular approach in functional data analysis to explore major sources of variation in a sample of random curves. These major sources of variation are represented by functional principal components (FPCs). Most existing FPCA approaches use a set of flexible basis functions such as B-spline basis to represent the FPCs, and control the smoothness of the FPCs by adding roughness penalties. However, the flexible representations pose difficulties for users to understand and interpret the FPCs. In this article, we consider a variety of applications of FPCA and find that, in many situations, the shapes of top FPCs are simple enough to be approximated using simple parametric functions. We propose a parametric approach to estimate the top FPCs to enhance their interpretability for users. Our parametric approach can also circumvent the smoothing parameter selecting process in conventional nonparametric FPCA methods. In addition, our simulation study shows that the proposed parametric FPCA is more robust when outlier curves exist. The parametric FPCA method is demonstrated by analyzing several datasets from a variety of applications.


The effect of processing fluency on impressions of familiarity and liking.
Processing fluency has been shown to have wide-ranging effects on disparate evaluative judgments, including judgments of liking and familiarity. One account of such effects is the hedonic marking hypothesis (Winkielman, Schwarz, Fazendeiro, & Reber, 2003), which posits that fluency is directly linked to affective preferences via a positive emotional reaction that is triggered by fluent processing. The evidence supporting this account suggests that fluency may exert a stronger influence on affective judgments than other judgments. The current study compared the effect of fluency on judgments of familiarity and liking. Contrary to predictions, liking judgments were not more strongly affected by fluency than familiarity judgments. In fact, the balance of the results showed the opposite pattern. When the type of judgment was manipulated between subjects (Experiment 1) or in a blocked design (Experiment 2), fluency had comparable effects on impressions of liking and familiarity. But when the type of judgment was manipulated in a mixed design (Experiments 3 and 6), or when both familiarity and liking judgments were given for all items (Experiments 4 and 5), only familiarity judgments were affected by the fluency manipulation. The dominance of the familiarity interpretation was found when fluency was manipulated artificially, via priming, and when inherent variations in fluency across the stimuli were considered. These results suggest that, within a given context, participants adopt a single interpretation for fluency, and the sense of familiarity that arises from fluent processing overshadows the sense of positivity, thus questioning aspects of the hedonic marking hypothesis.


Comparative efficacy and safety of different doses of ponatinib versus other tyrosine kinase inhibitors for the treatment of chronic myeloid leukemia: a systematic review and network meta-analysis.
Ponatinib was recommended with caution because of its high risk of causing arterial occlusion events in chronic myeloid leukemia (CML) patients. The purpose of this study was to understand the efficacy and safety of different doses of ponatinib in the treatment of CML, and to compare it with other tyrosine kinase inhibitors (TKIs). A network meta-analysis (NMA) was conducted by searching randomized controlled trials (RCTs) of ponatinib in patients with CML to compare the efficacy and safety of ponatinib, and ranked under the cumulative ranking curve (SUCRA) to evaluate the optimal treatment. A total of seven articles with eight RCTs were included in this study, involving 45 mg, 30 mg and 15 mg ponatinib doses. Seven outcome indexes were analyzed. The results showed that 45 mg ponatinib was superior to other doses of ponatinib and other TKIs in CCyR, MCyR and CHR, but the incidence of SAEs and AOEs was significantly higher than other treatment regimens. Ponatinib, with an initial dosage of 45 mg and a gradual reduction to 15 mg, may be a more favorable option for patients with CML at all stages of disease progression, rather than just those in the chronic phase of CML.


Effect of hepatitis C virus nonstructural protein NS3 on proliferation and MAPK phosphorylation of normal hepatocyte line.
To study the effect of hepatitis C virus nonstructural region 3 (HCV NS3) protein on proliferation and transformation of normal human liver cell line. QSG7701 cells were transfected with pRcHCNS3-5', pRcHCNS3-3' and pRcCMV using lipofectamine transfecting technique and selected with G418 method. Expression of HCV NS3 protein was determined by immunohistochemistry. Biologic characteristics of transfected cells were evaluated by population doubling time and soft agar assays. Activation of MAPK was analyzed using Western blot with phosphospecific monoclonal antibody against dually phosphorylated MAPK. QSG7701 cells transfected with pRcHCNS3-5' showed strong intracellular expression of HCVNS3 protein, and the positive signal was localized in cytoplasm. The expressing strength of HCVNS3 protein in pRcHCNS3-3'-transfected cells was weaker than that in pRcHCNS3-5'-transfected cells. The population doubling time in the transfected cells with pRcHCNS3-5' (12 h) was much shorter than those with pRcHCNS3-3', pRcCMV and normal cells (24, 26, 28 h, respectively) (P<0.01). The transfected cells with pRcHCNS3-5' showed much more anchorage independent colonies than that in those with pRcHCNS3-3' and pRcCMV (P<0.01). The cloning efficiencies of transfected cells with pRcHCNS3-5', pRcHCNS3-3', pRcCMV and controls were 33%, 1.33%, 1.46%, 1.11% respectively. The level of phosphorylated MAPK in the cells with pRcHCNS3-5' was much higher than that in those with pRcHCNS3-3'and pRcCMV and normal cells (P<0.01). The results suggest that (1) QSG7701 cells are a better human liver cell line for investigating the pathogenesis of HCV NS3 protein. (2) 5' region of the HCV genome segment encoding HCV NS3 is involved in cell growth and cell phenotype. (3) HCV NS3 N-terminal peptide may up-regulate the activation of MAPK, but not affect the expression of MAPK.


Association between cardiovascular risk-factors and venous thromboembolism in a large longitudinal study of French women.
Previous studies have shown conflicting results regarding the influence of cardiovascular risk-factors on venous thromboembolism. This study aimed to determine if these risk-factors, i.e. physical activity, smoking, hypertension, dyslipidaemia, and diabetes, were associated with the risk of venous thromboembolism, and to determine if these associations were confounded by BMI. We used data from the E3N cohort study, a French prospective population-based study initiated in 1990, consisting of 98,995 women born between 1925 and 1950. From the women in the study we included those who did not have prevalent arterial disease or venous thromboembolism at baseline; thus 91,707 women were included in the study. Venous thromboembolism cases were self-reported during follow-up, and verified via specific mailings to medical practitioners or via drug reimbursements for anti-thrombotic medications. Hypertension, diabetes and dyslipidaemia were self-reported validated against drug reimbursements or specific questionnaires. Physical activity, and smoking were based on self-reports. Cox-models, adjusted for BMI and other potential risk-factors were used to determine hazard ratios for incident venous thromboembolism. During 1,897,960 person-years (PY), 1, 649 first incident episodes of thrombosis were identified at an incidence rate of 0.9 per 1000 PY. This included 505 cases of pulmonary embolism and 1144 cases of deep vein thrombosis with no evidence of pulmonary embolism. Hypertension, dyslipidaemia, diabetes, smoking and physical activity were not associated with the overall risk of thrombosis after adjustment for BMI. Traditional cardiovascular risk factors were not associated with the risk of venous thromboembolism after adjustment for BMI. Hypertension, dyslipidaemia and diabetes may not be risk-factors for venous thromboembolism.


Effectiveness of somatosensory interventions on somatosensory, motor and functional outcomes in the upper limb post-stroke: A systematic review and meta-analysis.
Research mainly focuses on motor recovery of the upper limb after stroke. Less attention has been paid to somatosensory recovery. To review and summarize the effect of upper limb somatosensory interventions on somatosensory impairment, motor impairment, functional activity and participation after stroke. Biomedical databases Ovid Medline, EMBASE, Web of Science, PEDro, and OTseeker were searched with an update in May 2018. Randomized controlled trials investigating the effect of somatosensory-specific interventions focusing on exteroceptive, proprioceptive or higher cortical somatosensory dysfunction, or any combination were eligible for inclusion. Quality of included studies were assessed using Physiotherapy Evidence Database (PEDro) scale. Standardized Mean Differences and Mean Differences and 95% confidence intervals were calculated and combined in meta-analyses. Active somatosensory interventions did not show a significant effect on somatosensation and activity, but demonstrated a significant improvement in motor impairment (SMD = 0.73, 95% CI = 0.14 to 1.32). No study evaluating active somatosensory intervention included participation. Passive somatosensory interventions significantly improved light touch sensation (SMD = 1.13, 95% CI = 0.20 to 2.05). Passive somatosensory interventions did not show significant effects on proprioception and higher cortical somatosensation, motor impairment, activity and participation. To date, there is low quality evidence suggesting active somatosensory interventions having a beneficial effect on upper limb impairment and very low quality evidence suggesting passive somatosensory interventions improving upper limb light touch sensation. There is a need for further well-designed trials of somatosensory rehabilitation post stroke.


Monoclonal antibody localization of Lewis antigens in fixed tissue.
Monoclonal antibodies that bind specifically with Lewisa (Lea) and Lewisb (Leb) antigens were used in an immunoperoxidase assay to characterize Lewis (Le) antigenic profile of a variety of fixed tissues. Representative sections of normal and malignant tissue from the stomach, colon, pancreas, and kidney were examined and compared. Lea antigen was expressed more often in gastric adenocarcinomas (80%) than normal gastric mucosa (40%). In gastric tumors with concomitant expression of Le antigens, larger areas within an individual tumor expressed Lea antigen rather than Leb antigen. Expression of Leb antigen in normal colonic tissue was seen only in the proximal colon and not distal colon; colon carcinomas, on the other hand, expressed Leb antigen (64%) regardless of where the primary arose. Leb antigen was expressed in large pancreatic ducts (three of three) more often than Lea antigen (one of three); exclusive expression of Lea antigen was demonstrated in the proximal convoluted tubules of normal kidney (three of three) and in renal cell carcinomas (six of eight).


Improved glycaemic control with dipeptidyl peptidase-4 inhibition in patients with type 2 diabetes: vildagliptin (LAF237) dose response.
A novel treatment option for diabetic patients is the enhancement of incretin hormone activity by inhibition of the enzyme dipeptidyl peptidase-4 (DPP-4). This study was designed to establish a dose of the DPP-4-inhibitor vildagliptin (LAF237) that was effective in reducing HbA1c levels and was safe and well tolerated in patients with type 2 diabetes. The study of 279 patients with type 2 diabetes consisted of a 4-week run-in phase where patients received placebo and a 12-week active treatment phase where they received one of the following dosages of vildagliptin: 25 mg twice daily, 25, 50 or 100 mg once daily (qd), or placebo. There was a statistically significant reduction in HbA1c levels in the vildagliptin 50 mg qd (p=0.003) and 100 mg qd groups (p=0.004) compared with the placebo group. The mean 4-h postprandial glucose level was significantly reduced from placebo in the vildagliptin 50 mg qd group (p = 0.012) and mean 4-h postprandial insulin was significantly increased from baseline vs. placebo in the vildagliptin 100 mg qd group (p=0.022). The assessment of beta-cell function (HOMA-B) was significantly increased in the vildagliptin 100 mg qd treatment group (p=0.007). The incidence of adverse events was similar in all treatment groups including placebo. Vildagliptin, at 50 and 100 mg qd, was effective in reducing HbA1c levels compared with placebo in patients with type 2 diabetes. Vildagliptin at dosages up to 100 mg qd appeared safe and well tolerated.


Relevance of MRI for management of non-displaced lateral humeral condyle fractures in children.
The treatment for non-displaced (<2 mm displacement) fractures of the lateral humeral condyle in children is controversial. Most studies recommend non-surgical treatment. However, plain radiographs are not sufficient to evaluate extension of the fracture line through the articular cartilage. This explains the high frequency of secondary displacements and non-unions, despite well-conducted conservative treatment. We hypothesized that MRI could be used to analyse whether the fracture is complete or incomplete. This could help to determine whether surgical or conservative treatment is indicated. This prospective study enrolled children being treated for a non-displaced (< 2 mm gap) fracture of the lateral humeral condyle. All patients were treated with a long-arm cast in the emergency room. An MRI was done later on without sedation. A specific protocol was used to reduce the duration of the examination. T2-weighted and proton density fat-saturated sequences were used. Twenty-seven patients were enrolled: 16 boys and 11 girls with a mean age of 5 years (2-10). The MRI was performed an average of 7 days (1-23) after the fracture. The MRI could not be interpreted in two cases because the child had moved during the examination. In the other 25 patients, the fracture was incomplete in 17 patients and complete in 8 patients. Two children had secondary displacement diagnosed 7 and 11 days after the fracture event. These two patients underwent open reduction and internal fixation. There was no correlation between patient age and the fracture being complete or incomplete. There were no cases of non-union. MRI appears to be a reliable method for determining whether the fracture line is complete or incomplete. It can be performed without sedation, even in children as young as 2 years of age. Use of an injury-specific MRI protocol reduces the length of the examination, thereby improving its performance. We recommend that it be used to analyse non-displaced fractures of the lateral humeral condyle in children. 3 Prospective study.


Efficient and selective syntheses of (all-E)- and (6E,10Z)-2'-O-methylmyxalamides D via Pd-catalyzed alkenylation--carbonyl olefination synergy.
Highly efficient and selective syntheses of both (all- E) and (6 E,10 Z)-isomers of 2'- O-methylmyxalamide D ( 2 and 3), in which the crucial conjugated pentaene moieties were assembled in > or =98% stereoselectivity through the use of two Pd-catalyzed alkenylation reactions, the Horner-Wadsworth-Emmons (HWE) olefination, and either the Corey-Schlessinger-Mills modified (CSM-modified) Peterson olefination for 2 or the Still-Gennari olefination for 3, are reported. Either 2 or 3 was prepared in 16% yield in seven steps from propargyl alcohol.


Uncleaved 2A-peptide of foot-and-mouth disease virus can display foreign epitope-tag at the virion surface.
Foot-and-mouth disease virus (FMDV) capsid precursor protein P1-2A is cleaved by viral-encoded 3C protease (3Cpro) to generate VP0, VP3, VP1 and 2A proteins. It was reported earlier that substitution of a single amino acid residue within the 2A peptide sequence (L2P) blocked the 3Cpro mediated VP1/2A cleavage and produced 'self-tagged' FMDV particles containing uncleaved 2A-peptide. To determine whether the uncleaved 2A-peptide can function as a target structure to harbour and display exogenous epitope on FMDV particles, a full-length FMDV cDNA clone containing a HA-tag within the uncleaved 2A-peptide sequence was constructed. Subsequently, chimeric marker FMDV, displaying a HA-tag on the viral surface was rescued through reverse genetics approach. The 2A-HA epitope tag-inserted recombinant chimeric FMDV serotype O was genetically stable through up to ten serial passages in cell culture and exhibited growth properties similar to the parental virus. Furthermore the surface displayed HA-epitope tag was able to react with anti-HA antibodies as determined by various immuno-assays. The results from our study suggest that the uncleaved 2A-peptide of FMDV is suitable to present foreign antigenic epitopes on the surface of FMD virion.


[Occurrence and diagnostic relevance of virulence-associated factors in Streptococcus suis].
Streptococcus suis (Sc. suis) can cause very different clinical entities. In contrast to Sc. suis-associated pneumonia, the induction of meningitis, septicemia, and polyarthritis by certain Sc. suis strains requires the expression of virulence factors that contribute to the invasiveness of the pathogen. In the presented study, we examined the occurrence of known virulence-associated factors in Sc. suis isolates from samples sent to the Institute of Microbiology, School of Veterinary Medicine Hannover, in order to evaluate their significance as potential virulence factors in different disease complexes in Northern Germany. The results show that (i) MRP + EF + serotype 2 and MRP* EF-serotype 9 strains are statistically significant associated with the disease complex meningitis/septicemia/arthritis and, thus, have to be considered invasive strains, (ii) serotyping alone is not sufficient for identification of virulent strains, (iii) there is a remarkable heterogeneity among pneumonia-associated Sc. suis strains and (iv) activity of haemolysin or suilysin appears to be not appropriate as virulence marker. Finally, it has to be noted that at present only half of the Sc. suis isolates from pigs with meningitis/septicemia/poyarthritis can be characterised by the detection of virulence-associated factors. Thus, the identification and characterisation of additional, serotype independent virulence factors of Sc. suis is a very important issue in future studies.


Comparing brain connectivity metrics: a didactic tutorial with a toy model and experimental data.
The objective of this paper is to didactically compare resting state connectivity networks computed using two different methods called phase locking value (PLV) and convergent cross-mapping (CCM). PLV is a ubiquitous measure of connectivity in electrophysiological research but is less often applied to fMRI BOLD timeseries since this model-based metric assumes that oscillatory coupling is a sufficient condition for connectivity. Alternatively, CCM is a model-free method, which detects potentially nonlinear causal influences based on the ability to estimate one timeseries with another and does not assume an oscillatory structure. We use a toy dataset to test the PLV and CCM algorithms under different known synchronization conditions. Additionally, experimental resting state EEG and fMRI datasets are used for comparison. The results show that the resting state brain networks computed using both algorithms produce similar results for both resting state EEG and fMRI datasets. For both neuroimaging datasets, the network characteristics follow the same trends and the similarity between the computed networks, for both algorithms, is highly significant. CCM is able to identify low or one-way connection strengths better than PLV but takes exponentially longer to compute. Based on these results, PLV provides a good metric for on-line network identification because it is both computationally fast and an excellent approximation of the network computed with CCM.


A note on lactobacilli and beta-glucuronidase activity in the intestinal contents of mice.
A comparison was made of caecal beta-glucuronidase activity in the caecal contents of mice with or without lactobacilli as members of the normal microflora. Male mice that did not have lactobacilli as intestinal inhabitants had approximately 52% more caecal beta-glucuronidase activity than their female counterparts. Colonization of male mice by lactobacilli reduced the beta-glucuronidase activity to that of female mice.


Instrumented Gait Analysis: A Tool in the Treatment of Spastic Gait Dysfunction.
Spasticity is a term commonly used to describe a collection of muscle overactivity patterns associated with the upper motor neuron syndrome including actual spasticity, clonus, dystonia, co-contraction, associated reactions, and flexor or extensor spasms. Gait dysfunction in the upper motor neuron syndrome can be due to a combination of paresis, impaired coordination and balance, and muscle overactivity and contracture. Treatment options include physiotherapy, assistive devices, orthotic devices, oral and intrathecal medications, intramuscular chemodenervation, neurolysis, and/or neuro-orthopaedic surgical procedures. The objective evaluation of walking and its underlying muscle activation patterns can be performed by the simultaneous collection of joint kinematics, kinetics, and dynamic electromyography (EMG) data that serve as an extension of the physical examination to better discern primary gait deviations from compensatory gait deviations as well as underlying muscle overactivity from contracture. Despite the science behind instrumented three-dimensional gait analysis, its specific contribution to clinical and surgical decision-making is not well utilized because of its associated cost, the incorrect view by some insurance companies of gait analysis being a research technique, the dearth of qualified clinical gait and motion analysis laboratories accessible to adult patients, and limited access to orthopaedic surgeons who have experience in the treatment of neurological disorders for this patient population.


Cluster ion beam profiling of organics by secondary ion mass spectrometry--does sodium affect the molecular ion intensity at interfaces?
The use of cluster ion beam sputtering for depth profiling organic materials is of growing technological importance and is a very active area of research. At the 44th IUVSTA Workshop on "Sputtering and Ion Emission by Cluster Ion Beams", recent results were presented of a cluster ion beam depth profile of a thin organic molecular layer on a silicon wafer substrate. Those data showed that the intensity of molecular secondary ions is observed to increase at the interface and this was explained in terms of the higher stopping power in the substrate and a consequently higher sputtering yield and even higher secondary ion molecular sputtering yield. An alternative hypothesis was postulated in the workshop discussion which may be paraphrased as: "under primary ion bombardment of an organic layer, mobile ions such as sodium may migrate to the interface with the inorganic substrate and this enhancement of the sodium concentration increases the ionisation probability, so increasing the molecular ion yield observed at the interface". It is important to understand if measurement artefacts occur at interfaces for quantification as these are of great technological relevance - for example, the concentration of drug in a drug delivery system. Here, we evaluate the above hypothesis using a sample that exhibits regions of high and low sodium concentration at both the organic surface and the interface with the silicon wafer substrate. There is no evidence to support the hypothesis that the probability of molecular secondary ion ionisation is related to the sodium concentration at these levels.


Human and murine obliterative bronchiolitis in transplant.
Obliterative bronchiolitis is a devastating illness that limits the long-term success of lung transplantation. Its high prevalence and overall poor response to current therapeutic measures demands further research to elucidate pathogenic mechanisms. Toward this goal, there is a role for animal models to study the mechanisms of obliterative bronchiolitis, such as the murine heterotopic tracheal allograft model. This review compares the tracheal allograft model to human obliterative bronchiolitis pathology and highlights the important mechanisms of airway rejection described using this model. Although certain limitations exist, the pursuit of proof-of-concept studies in this model, as well as other animal models, can provide the basis for genetic and cellular translational human studies directed toward post-transplant obliterative bronchiolitis pathogenesis. To meet these challenges, we call for the establishment of a National Institutes of Health-supported Lung Transplant Network to better orchestrate translational research efforts in obliterative bronchiolitis pathogenesis and treatment, and to advance the field of lung transplantation.


Abnormal macrophages and NK cell cytotoxicity in human systemic lupus erythematosus and the role of interferon and serum factors.
Macrophage (MO) and natural killer (NK) cell mediated cytotoxicity to K562 target cells were strikingly decreased in patients with systemic lupus erythematosus (SLE). SLE NK cells failed to release soluble factor(s) for lysing the targets. IFN-induced enhancement of both types of cytotoxicity was impaired. NK cells from healthy subjects kept their activity in culture with or without IFN for more than six days whereas SLE NK cell activity declined to zero at day 3. So, the increased IFN level of many SLE patients and a possible prior IFN priming effect seemed unrelated to the insensitivity to exogenous IFN in vitro. Inhibition factor(s) of SLE serum suppressed NK cytotoxicity in the presence of IFN whereas IFN sensitivity of MO remained unaffected indicating the complex regulation by serum components of immune reactions.


Identification and frequency estimation of inversion polymorphisms from haplotype data.
Structural rearrangements, including copy-number alterations and inversions, are increasingly recognized as an important contributor to human genetic variation. Copy number variants are readily measured via array-based techniques like comparative genomic hybridization, but copy-neutral variants such as inversion polymorphisms remain difficult to identify without whole genome sequencing. We introduce a method to identify inversion polymorphisms and estimate their frequency in a population using readily available single nucleotide polymorphism (SNP) data. Our method uses a probabilistic model to describe a population as a mixture of forward and inverted chromosomes and identifies putative inversions by characteristic differences in haplotype frequencies around inversion breakpoints. On simulated data, our method accurately predicts inversions with frequencies as low as 25% in the population and reliably estimates inversion frequencies over a wide range. On the human HapMap Phase 2 data, we predict between 88 and 142 inversion polymorphisms with frequency ranging from 20 to 81 percent. Many of these correspond to known inversions or have other evidence supporting them, and the predicted inversion frequencies largely agree with the limited information presently available.


A near-infrared photodetector based on carbon nanotube transistors exhibits ultra-low dark current through field-modulated charge carrier transport.
Near-infrared photodetectors (NIR PDs) are devices that convert infrared light signals, which are widely used in military and civilian applications, into electrical signals. However, a common problem associated with PDs is a high dark current. Interestingly, gate voltage can regulate carrier migration in the channels. In this study, a PbS quantum dot heterojunction combined with a carbon nanotube (CNT) field effect transistor (FET) is designed and described. Significantly, this NIR PD achieves field-modulated carrier transport in a CNT transistor, in which the dark current is effectively regulated by the gate voltage. In this PD, an ultra-low dark current of 8 pA is obtained by gate voltage regulation. Moreover, the device shows a fast response speed of 6.5 ms and a high normalized detectivity of 4.75 × 1011 Jones at 0.085 W cm-2 power density and -0.2 V bias voltage. Overall, this work details a novel strategy for the fabrication of a PD with an ultra-low dark current based on a FET.


General consensus on multimodal functions and validation analysis of perinatal derivatives for regenerative medicine applications.
Perinatal tissues, such as placenta and umbilical cord contain a variety of somatic stem cell types, spanning from the largely used hematopoietic stem and progenitor cells to the most recently described broadly multipotent epithelial and stromal cells. As perinatal derivatives (PnD), several of these cell types and related products provide an interesting regenerative potential for a variety of diseases. Within COST SPRINT Action, we continue our review series, revising and summarizing the modalities of action and proposed medical approaches using PnD products: cells, secretome, extracellular vesicles, and decellularized tissues. Focusing on the brain, bone, skeletal muscle, heart, intestinal, liver, and lung pathologies, we discuss the importance of potency testing in validating PnD therapeutics, and critically evaluate the concept of PnD application in the field of tissue regeneration. Hereby we aim to shed light on the actual therapeutic properties of PnD, with an open eye for future clinical application. This review is part of a quadrinomial series on functional/potency assays for validation of PnD, spanning biological functions, such as immunomodulation, anti-microbial/anti-cancer, anti-inflammation, wound healing, angiogenesis, and regeneration.


Epidemiological survey of swine cysticercosis in two rural communities of West-Cameroon.
To determine the prevalence of porcine cysticercosis, a survey was carried out in 27 villages belonging to two rural communities of West-Cameroon (Bafou and Bamendou). Between January and August, 2000, a total of 707 pigs were examined serologically and by tongue inspection. Serum samples were examined for circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked-immunosorbent assay (Ag-ELISA) and for antibodies against cysticerci (Ab-ELISA). Seventy eight samples (11.0%) were found positive in the Ag-ELISA and 154 (21.8%) in the Ab-ELISA, while by tongue inspection on the same animals cysticerci were detected in 43 pigs (6.1%). Gibbs sampling using results of these three tests indicated that the estimated prevalence of porcine cysticercosis was 10.9%. Analysis of the Ag-ELISA results demonstrated that adult pigs showed a significantly higher seroprevalence (15%) than young ones (8.4%). There was no statistical difference in cysticercosis prevalence in pigs raised in households with or without a latrine. Animals that were reported to be usually confined were significantly less infected (9.9%) than free-roaming pigs (16.2%). Infection rates were significantly higher in pigs that had access to human faeces (13.8%) than those which did not have access (9.1%). This study has identified some community behavioural and environmental practices that should be modified to prevent continuous transmission of porcine cysticercosis.


Pre-exposure chemoprophylaxis (PREP) as an HIV prevention strategy.
Chemoprophylaxis may be a prevention strategy for the sexual transmission of human immunodeficiency virus (HIV). Evidence suggests that condom use has waned with the availability of antiretroviral medication, at least in some resource-rich settings. Barrier methods of HIV prevention have inherent problems, and the potential for failure. Microbicide research has focused primarily on male-to-female transmission. Analogous to post-exposure prophylaxis, HIV prevention may be achieved by pre-exposure prophylaxis in some settings. Research in this potential strategy may be rewarding.


Involvement of p300 in TGF-beta/Smad-pathway-mediated alpha2(I) collagen expression in mouse mesangial cells.
Transforming growth factor beta 1 (TGF-beta1) induces alpha2(I) collagen gene (COL1A2) expression in mesangial cells through physical and functional cooperation of Smad proteins and Sp1. A transcriptional coactivator, p300, is also suggested to play an important role in TGF-beta1/Smad signal transduction. However, the role of p300 in TGF-beta1/Smad-pathway-mediated transcriptional activation of the COL1A2 gene in mesangial cells is still obscure. Endogenous p300 expression and its modulation by TGF-beta1 were evaluated by Western blotting and immunofluorescence. The physical interaction of p300 with Smad2/3 was examined by immunoprecipitation followed by Western blotting. The functional role of p300 in TGF-beta1/Smad-pathway-mediated COL1A2 transcription was investigated in cotransfection experiments using a COL1A2 promoter-luciferase reporter gene construct and p300 expression plasmids. TGF-beta1 induced COL1A2 gene expression in cultured mouse mesangial cells which was blocked by overexpression of inhibitory Smad7. In addition, TGF-beta1-induced nuclear export of endogenous Smad7 was observed in mouse mesangial cells. Endogenous p300 was expressed in the nucleus of the cells. TGF-beta1 induced interaction of endogenous p300 with Smad2/3, and a dominant negative construct of p300 inhibited the TGF-beta1-induced COL1A2 expression in cultured mouse mesangial cells. p300 may be involved in TGF-beta1/Smad-pathway-mediated type I collagen gene transcription in mouse mesangial cells. Our findings would reveal a molecular basis of TGF-beta1-induced type I collagen gene transcription in mouse mesangial cells.


Diabetes and ageing-induced vascular inflammation.
Diabetes and the ageing process independently increase the risk for cardiovascular disease (CVD). Since incidence of diabetes increases as people get older, the diabetic older adults represent the largest population of diabetic subjects. This group of patients would potentially be threatened by the development of CVD related to both ageing and diabetes. The relationship between CVD, ageing and diabetes is explained by the negative impact of these conditions on vascular function. Functional and clinical evidence supports the role of vascular inflammation induced by the ageing process and by diabetes in vascular impairment and CVD. Inflammatory mechanisms in both aged and diabetic vasculature include pro-inflammatory cytokines, vascular hyperactivation of nuclear factor-кB, increased expression of cyclooxygenase and inducible nitric oxide synthase, imbalanced expression of pro/anti-inflammatory microRNAs, and dysfunctional stress-response systems (sirtuins, Nrf2). In contrast, there are scarce data regarding the interaction of these mechanisms when ageing and diabetes co-exist and its impact on vascular function. Older diabetic animals and humans display higher vascular impairment and CVD risk than those either aged or diabetic, suggesting that chronic low-grade inflammation in ageing creates a vascular environment favouring the mechanisms of vascular damage driven by diabetes. Further research is needed to determine the specific inflammatory mechanisms responsible for exacerbated vascular impairment in older diabetic subjects in order to design effective therapeutic interventions to minimize the impact of vascular inflammation. This would help to prevent or delay CVD and the specific clinical manifestations (cognitive decline, frailty and disability) promoted by diabetes-induced vascular impairment in the elderly.


Bispecific antibodies targeting CTLA-4: game-changer troopers in cancer immunotherapy.
Antibody-based cancer immunotherapy has become a powerful asset in the arsenal against malignancies. In this regard, bispecific antibodies (BsAbs) are a ground-breaking novel approach in the therapy of cancers. Recently, BsAbs have represented a significant advancement in improving clinical outcomes. BsAbs are designed to target two different antigens specifically. Over a hundred various BsAb forms currently exist, and more are constantly being manufactured. An antagonistic regulator of T cell activation is cytotoxic T lymphocyte-associated protein 4 (CTLA-4) or CD152, a second counter-receptor for the B7 family of co-stimulatory molecules was introduced in 1996 by Professor James P. Allison and colleagues. Contrary to the explosive success of dual immune checkpoint blockade for treating cancers, a major hurdle still yet persist is that immune-related adverse events (irAEs) observed by combining immune checkpoint inhibitors (ICIs) or monoclonal antibodies such as ipilimumab (anti-CTLA-4) and nivolumab (anti-PD-1). A promising strategy to overcome this hurdle is using BsAbs. This article will summarize BsAbs targeting CTLA-4, their applications in cancer immunotherapy, and relevant clinical trial advances. We will also discuss the pre-clinical rationale for using these BsAbs, and provide the current landscape of the field.


Noninvasive evaluation of endothelial vascular reactivity: should the quest continue?
Endothelial dysfunction is an early event in the continuum of the pathogenesis of atherosclerosis that persists throughout the disease process. Endothelial dysfunction may be a valuable marker or barometer of the composite effect of the various predisposing factors, even some yet unknown, on the subsequent risk of cardiovascular events. Accumulating data suggest that clinical evaluation of endothelial function may provide incremental value to current risk stratification and to the tailoring of pharmacological and nonpharmacological therapy. This has prompted many investigators to probe the integrity of endothelium-dependent vascular reactivity in an effort to uncover clinically useful methods of evaluating endothelial function. For practical purposes, reactive hyperemia of the forearm has become the most popular stimulus for testing vascular reactivity, and several methods have been developed to quantify this response and study its determinants. High-resolution brachial ultrasound and venous impedance plethysmography are used extensively in mechanistic studies; however, for practical purposes, it is unlikely that these methods could be used in routine clinical practice. Therefore, some alternatives are being actively developed. The greatest remaining challenge is to find firm evidence that endothelial function can be used as a clinical surrogate of cardiovascular events. Validation of these methods and their evaluation in large-scale trials is needed to determine whether abnormal vascular reactivity is indeed a good barometer of cardiovascular risk.


[Percutaneous cooled-tip microwave ablation under ultrasound guidance for primary liver cancer: analysis of major complications in 693 patients].
To analyze the major complications of percutaneous cooled-tip microwave ablation for the treatment of primary liver cancer and the possible risk factors of severe complications in a series of 693 patients. The clinicopathological data of 693 patients with primary liver cancer who underwent ultrasound-guided percutaneous cooled-tip microwave (MW) ablation in our hospital over the past 5 years were retrospectively analyzed, and the risk factors of severe complications were explored. In a total of 693 patients with 898 primary liver tumors were treated and 1111 MW ablation sessions were performed. The mean diameter of tumors was (2.5 ± 1.2) cm and the range was 0.4 - 10.0 cm. Three deaths occurred in the peri-ablation period, including one case died of multiorgan failure, one case died of pulmonary embolism and one case died of hepatorenal syndrome. Major complications occurred in 27 (3.9%) patients, including 12 pleural effusion requiring thoracentesis (1.7%), 10 tumor seeding (1.4%), 3 liver abscess and empyema (0.4%), 1 hemorrhage requiring arterial embolization (0.1%), and 1 bile duct injury (0.1%). The Chi-square test results showed that the diameter of tumors, number of MW ablation sessions and histological type of tumor were significantly associated with the major complications rate (P < 0.05). The multiple variables 1ogistic regression analysis showed that only type of tumors was associated with the major complication rate (P < 0.05). Results of this study confirm that cooled-tip MW ablation is a relatively low-risk and effective minimally invasive procedure for the treatment of primary liver cancer. Proper direction for the treatment of cholangiocarcinoma (ICC) patients as well as fewer ablated tumor numbers during one hospital stay may help minimize the major complication rate in patients with primary liver cancer treated by ultrasound-guided percutaneous cooled-tip microwave ablation.


Direct force measurements of insulin monomer-monomer interactions.
Direct measurement of the forces involved in protein-protein and protein-receptor interactions can, in principle, provide insight necessary for the advancement of structural biology, molecular biology, and the development of therapeutic proteins. The protein insulin is illustrative in this respect as the mechanisms of insulin dimer dissociation and insulin-insulin receptor binding are crucial to the efficacy of insulin medications for the control of diabetes. Insulin molecules, modified with a photochemically active azido functionality on specific residues, were attached to force microscope tips and opposing mica surfaces in configurations that would either favor or disfavor dimer formation. Force curve measurements performed in buffer solution revealed the complexity of the insulin monomer-monomer interaction with multiple unbinding events occurring upon tip retraction, suggesting disruption of discrete molecular bonds at the monomer-monomer interface. Furthermore, the force curves exhibit long-range unbinding events, consistent with considerable elongation of the insulin molecule prior to dissociation. The unbinding forces observed in this study are the result of a combination of molecular disentanglement and dimer dissociation processes.


Prospective study evaluating total disc replacement: preliminary results.
For many years, there has been an interest in disc replacements. There are a few retrospective studies from Europe reporting promising results for these devices. The purpose of this study is to review the preliminary results of the SB Charité disc replacement device in a prospective study. The study group consists of a consecutive series of our first 57 patients who received the Link SB Charité device implanted through a mini-open anterior approach. The primary clinical indications for disc replacement were single-level symptomatic disc disease at L4-L5 or L5-S1 and no significant facet joint changes. The outcome data include the Visual Analog Scale (VAS) assessing pain intensity and the Oswestry Low Back Pain Disability Questionnaire. Data were collected prior to surgery and at 6 weeks, 3 months, 6 months, and 12 months after surgery. Twenty-four-month follow-up data are currently being collected. The mean VAS score improved 50.8% by the 6-week follow-up, and significant improvement was maintained throughout follow-up. The mean Oswestry score improved 38.7% at the 6-week follow-up, and further slight improvement was noted during the later follow-up periods. This study found that significant improvements were achieved by the 6-week postoperative visit in the pain and disability scores following disc replacement surgery. Significant improvement was maintained through the 12-month follow-up visit. Although these preliminary results are encouraging, long-term follow-up and a greater number of patients are needed to further evaluate the outcome of these devices.


Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors.
Structural studies of human G protein-coupled receptors (GPCRs) have recently been accelerated through the use of a fusion partner that was inserted into the third intracellular loop. Using chimeras of the human β(2)-adrenergic and human A(2A) adenosine receptors, we present the methodology and data for the initial selection of an expanded set of fusion partners for crystallizing GPCRs. In particular, use of the thermostabilized apocytochrome b(562)RIL as a fusion partner displays certain advantages over previously utilized fusion proteins, resulting in a significant improvement in stability and structure of GPCR-fusion constructs.


GPER-independent inhibition of adrenocortical cancer growth by G-1 involves ROS/Egr-1/BAX pathway.
We previously demonstrated that treatment of the H295R adrenocortical cancer cell line with the non-steroidal, high-affinity GPER (G protein-coupled estrogen receptor 1) agonist G-1 reduced tumor growth in vitro and in vivo through a GPER independent action. Moreover, we observed that G-1 treatment induces cell-cycle arrest and apoptosis following a sustained ERK1/2 activation. However, the precise mechanisms causing these effects were not clarified. Starting from our preliminary published results, we performed a microarray study that clearly evidenced a strong and significative up-regulation of EGR-1 gene in H295R cells treated for 24h with micromolar concentration of G-1. The microarray findings were confirmed by RT-PCR and Western-blot analysis as well as by immunofluorescence that revealed a strong nuclear staining for EGR-1 after G-1 treatment. EGR-1 is a point of convergence of many intracellular signaling cascades that control tumor cell growth and proliferation as well as others that relate to cell death machinery. Here we found that the increased Egr-1 expression was a consequence of G-1-mediated ROS-dependent ERK activation that were promptly reversed by the presence of the antioxidant n-acetyl-cysteine. Finally, we observed that silencing EGR-1 gene expression reversed the main effects induced by G-1 in ACC cells, including upregulation of the negative regulator of cell cycle, p21Waf1/Cip1 and the positive regulator of mitochondrial apoptotic pathway, BAX, as well as the cell growth inhibition. The identified ROS/MAPK/Egr-1/BAX pathway as a potential off-target effect of the G-1 could be useful in implementing the pharmacological approach for ACC therapy.


[Relationship between hemorheology and pro- and anti-coagulation factors in inflammatory and degenerative vascular diseases].
After having found out an increase in the Von Willebrand Factor (vWF) and a relation between its persistance and seriousness of some vasculities, we looked for a link between changes in vWF level and hemorheology, their role and interdependence in the evolution of the vascular diseases, in complications and in relapses. We studied patients affected with degenerative and inflammatory vascular diseases and we determined rheologic parameters, in traditional and personal innovative way, pro- and anti-coagulation factors, other inflammatory proteins and hematic parameters. As regards coagulation only the vWF appeared increased in a significant way, without differences between two types of vascular diseases. As regards hemorheology, in both of them we measured an increase in the whole blood viscosity but with a different behaviour of its components. In the degenerative vascular diseases there was an increase both in corpuscular viscosity and in the plasma-erythrocyte one, a no significant increase in the plasma viscosity whereas the erythrocyte deformability was in the normal range. In vasculitis there was the highest increase in the plasma and corpuscular viscosities with a decrease in the deformability without changes in the plasma-erythrocyte viscosity. We give some interpretations on their different behaviour and a right meaning of the involved phenomena in every kind of rheologic parameter.


In vitro culture of human peritoneal mesothelium for investigation of mesothelial dysfunction during peritoneal dialysis.
There is a growing need for more biocompatible peritoneal dialysis fluid formulations. Using a method of in vitro cell culture we have examined the effect of osmotic agents, antiseptics and drug additives on various functions of human peritoneal mesothelial cells (HPMC). We have demonstrated that glucose and amino acids reduce the HPMC proliferation rate in a dose-dependent manner. 3H-methyl-thymidine incorporation into HPMC exposed to either 1.1% amino acid mixture or 2.0% glucose was decreased to (mean +/- SD) 32.5 +/- 6.1% and 16.8 +/- 1.2% of the control level respectively. Cultured HPMC have been shown to produce a fibrinolytic activity which may be estimated by the rate of plasma clot lysis. This activity may be diminished following the pre-incubation with 2.0% glucose and 1.1% amino acid mixture. We have found that povidone-iodine exerts a dose-dependent cytotoxic effect towards HPMC as measured by the increased 86Rb or 3H-inulin release from radiolabelled cells. We have also demonstrated that insulin enhances the 86Rb uptake and stimulates the Na, K-ATPase activity in HPMC but on the other hand is capable of reducing the phospholipids secretion from HPMC. Glucose, hydrocortisone or verapamil have also been shown to inhibit the release of mesothelial phospholipids. These data indicate that the mesothelial cell culture provides a convenient model for testing the biocompatibility of peritoneal dialysis solutions and assessing the dysfunction of mesothelial cells during peritoneal dialysis.