Bergenin Inhibits Tumor Growth and Overcomes Radioresistance by Targeting Aerobic Glycolysis.
Hexokinase 2 (HK2), the first glycolytic rate-limiting enzyme, is closely correlated with the occurrence and progression of tumors. Effective therapeutic agents targeting HK2 are urgently needed. Bergenin has exhibited various pharmacological activities, such as antitumor properties. However, the effects of bergenin on the abnormal glucose metabolism of cancer cells are yet unclear. In this study, HK2 was overexpressed in OSCC tissues, and the depletion of HK2 inhibited the growth of OSCC cells in vitro and in vivo. Moreover, these results showed that the natural compound, bergenin, exerted a robust antitumor effect on OSCC cells. Bergenin inhibited cancer cell proliferation, suppressed glycolysis, and induced intrinsic apoptosis in OSCC cells by downregulating HK2. Notably, bergenin restored the antitumor efficacy of irradiation in the radioresistant OSCC cells. A mechanistic study revealed that bergenin upregulated the protein level of phosphatase and the tensin homolog deleted on chromosome 10 (PTEN) by enhancing the interaction between PTEN and ubiquitin-specific protease 13 (USP13) and stabilizing PTEN; this eventually inhibited AKT phosphorylation and HK2 expression. Bergenin was identified as a novel therapeutic agent against glycolysis to inhibit OSCC and overcome radioresistance. Targeting PTEN/AKT/HK2 signaling could be a promising option for clinical OSCC treatment.


Silencing of microRNA-21 in vivo ameliorates autoimmune splenomegaly in lupus mice.
MicroRNAs (miRNAs) have been implicated in B cell lineage commitment, regulation of T cell differentiation, TCR signalling, regulation of IFN signalling, and numerous other immunological processes. However, their function in autoimmunity, and specifically in systemic lupus erythematosus (SLE), remains poorly understood. B6.Sle123 is a spontaneous genetic mouse model of SLE characterized by autoantibody production, lymphosplenomegaly, and glomerulonephritis. We identified several differentially regulated miRNAs in B and T lymphocytes of B6.Sle123 mice. We found that miR-21 expression in lupus B and T cells is up-regulated and that in vivo silencing of miR-21 using a tiny seed-targeting LNA reversed splenomegaly, one of the cardinal manifestations of autoimmunity in B6.Sle123 mice, and de-repressed PDCD4 expression in vivo and in vitro. In addition, treatment with anti-miR-21 altered CD4/CD8 T cell ratios and reduced Fas receptor-expressing lymphocyte populations. Our study shows that tiny LNAs can be used to efficiently antagonize endogenous miRNAs in peripheral lymphocytes in vivo and in primary lymphocytes cultured ex vivo and can alter the course of a spontaneous genetic disease in mice.


Preventing Healthcare-Associated Infections by Monitoring the Cleanliness of Medical Devices and Other Critical Points in a Sterilization Service.
It is well known that the common goal of all central sterile supply departments (CSSDs) is to prevent healthcare-associated infections. Such infections entail high costs to society, not only economic but also social. Therefore, delivering safe medical devices and guaranteeing a positive contribution to the control of healthcare-associated infections form the main responsibilities of a CSSD. The monitoring of the effectiveness of medical device cleaning processes is highly recommended. However, ensuring a flawless environment for the preparation, assembly, and packaging of medical devices and clean handling of sterilized items is crucial to achieving the goal of safe medical devices. This study analyzed not only the cleanliness of surgical instruments but also two critical aspects of the surrounding environment: the cleanliness of work surfaces and the cleanliness of workers' hands. To evaluate the cleanliness of surgical instruments, two methods were used: the adenosine triphosphate (ATP) detection method and a residual protein test. It was not the intention of this work to make an exhaustive comparison of these methods. The ATP bioluminescence method was also used for monitoring the cleanliness of work surfaces and workers' hands. The aims of this study were to establish the most suitable method of evaluating the cleanliness of reusable medical devices in the CSSD and to assess the quality of the environment. Assessing the surgical instruments, work surfaces, and staff hands for cleanliness allowed the identification of possible contamination sources and to correct them by improving cleaning/disinfection protocols. Furthermore, the use of ATP monitoring tests of workers' hands highlighted the importance of staff compliance with good practice guidelines. Thus, these results have a positive impact on the CSSD quality system and, consequently, on patient safety.


Answering some questions about structured illumination microscopy.
Structured illumination microscopy (SIM) provides images of fluorescent objects at an enhanced resolution greater than that of conventional epifluorescence wide-field microscopy. Initially demonstrated in 1999 to enhance the lateral resolution twofold, it has since been extended to enhance axial resolution twofold (2008), applied to live-cell imaging (2009) and combined with myriad other techniques, including interferometric detection (2008), confocal microscopy (2010) and light sheet illumination (2012). Despite these impressive developments, SIM remains, perhaps, the most poorly understood 'super-resolution' method. In this article, we provide answers to the 13 questions regarding SIM proposed by Prakash et al. along with answers to a further three questions. After providing a general overview of the technique and its developments, we explain why SIM as normally used is still diffraction-limited. We then highlight the necessity for a non-polynomial, and not just nonlinear, response to the illuminating light in order to make SIM a true, diffraction-unlimited, super-resolution technique. In addition, we present a derivation of a real-space SIM reconstruction approach that can be used to process conventional SIM and image scanning microscopy (ISM) data and extended to process data with quasi-arbitrary illumination patterns. Finally, we provide a simple bibliometric analysis of SIM development over the past two decades and provide a short outlook on potential future work. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 2)'.


Impact of Chocolate percutaneous transluminal angioplasty balloon on vessel preparation in drug-coated balloon angioplasty for femoropopliteal lesion.
To compare the impact of Chocolate and conventional balloons on vessel preparation in percutaneous transluminal angioplasty. This single-center retrospective study included 111 endovascular therapy consecutive cases of femoropopliteal lesions using drug-coated balloon strategy with a 1:1 pre-dilation balloon diameter between February 2020 and August 2021, divided into the Chocolate percutaneous transluminal angioplasty (n = 48) and conventional (n = 63) groups. Before the availability of Chocolate balloons in Japan (December 2020), a standard semi-compliant or non-compliant balloon was used for vessel preparation. The primary endpoint was rate of severe dissection after pre-dilatation. Secondary endpoints were angiographic percent diameter stenosis, bailout stent rate, primary patency rate, and freedom from target-lesion-revascularization rate at six months. There was no significant difference in patient and lesion characteristics. The procedural characteristics comprised balloon length 90 ± 37 and 149 ± 95 mm (P = 0.004) and inflation pressure 11 ± 3 and 16 ± 7 atm (P < 0.001) in the Chocolate and conventional groups, respectively. Regarding primary endpoint, rates of severe dissection were 4.2% and 25% (P = 0.003); regarding secondary endpoints, percent diameter stenosis was 18 ± 15% and 20 ± 17% (P = 0.409), and the rate of bailout stenting was 2.1% and 15.9% (P = 0.016) in the Chocolate and conventional groups, respectively. The primary patency rates at six months were 89.1% and 85.2% (P = 0.670), and freedom from target-lesion-revascularization rate at six months was 100% and 92.8% (P = 0.691) in the Chocolate and conventional groups, respectively. Chocolate percutaneous transluminal angioplasty balloons reduce the rate of severe dissection while maintaining a sufficient dilatation effect during drug-coated balloon vessel preparation.


Mechanical Thrombectomy for Acute Stroke in Patients with Target and Unrelated Vessel AVMs and AVFs: A Case Series.
Advances in acute and long-term poststroke care have resulted in improved survival and functional outcomes for patients who have suffered large vessel ischemic strokes. For years, tissue plasminogen activator was the mainstay of treatment for acute stroke. Its use was previously limited to patients without known comorbid intracranial vascular pathology because of concern for bleeding risk. More recently, however, the use of tissue plasminogen activator in select patients with vascular anomalies has increased and is now largely thought to be safe. With the safety and efficacy of mechanical thrombectomy now proven for large vessel occlusions (LVOs), similar investigation is needed to assess procedural safety in patients with concomitant arteriovenous (AV) malformations or fistulae. We reviewed patients treated for LVOs at our institution and those of our collaborators and identified 6 patients who were treated for LVO with either known or incidentally identified concomitant AV malformations or dural AV fistulae. We present a case series of 6 patients with nonaneurysmal intracranial vascular lesions who underwent mechanical thrombectomy for LVO without complications related to these lesions. Although limited by small size, our series adds to the literature evidence that mechanical thrombectomy for LVO can safely be performed with concomitant dural AV fistulae and AV malformations.


Strategies for insulin initiation: insights from the French LIGHT observational study.
The progressive nature of type 2 diabetes necessitates exogenous insulin use for most patients; basal insulin plus oral anti-diabetes drugs (OADs) is a well-validated way to facilitate insulin initiation. The primary aim of this study was to explore insulin initiation strategies and outcomes for patients using insulin detemir or glargine plus oral anti-diabetes drugs. LIGHT was a 3-month, longitudinal observational study conducted across 761 French centres in insulin-naïve type 2 diabetes patients managed under routine clinical care conditions, in either primary or secondary care. Endpoints included changes in HbA(1c) , fasting plasma glucose (FPG), rate of hypoglycaemia, weight, and adverse events. Most physicians initiated a basal analogue to improve glycaemic control (97%), with many delaying beginning treatment for several months (9 ± 9.0 months for general practitioners, 10.2 ± 16.2 months for specialists). Most patients continued oral anti-diabetes drug therapy (95%) and lifestyle measures (92%), with 2-3 blood glucose readings per day and follow-up telephone calls for dose optimization. Mean change in HbA(1c) from baseline was - 1.3%, and - 3.1 mmol/L for fasting plasma glucose (both p < 0.0001). Hypoglycaemia increased from 1.4 to 5.6 events/patient/year (p < 0.0001), and weight decreased on average by 0.5 kg with detemir, with no change in glargine. Most patients (93%) reported being satisfied or very satisfied with their insulin. Insulin initiation with detemir or glargine can be successfully managed in both primary and secondary care; the benefits of basal analogues (once-daily dosing, low rates of hypoglycaemia compared with neutral protamine Hagedorn) may have contributed to patient acceptance of the regimen.


Lupus nephritis associated with CD20+ B-cell NHL.
The combination of NHL and documented malignancy-associated glomerulonephritis is uncommon. Also, no single renal pathological entity is consistently found in patients with NHL. Epstein-Barr virus (EBV) infection may manifest as systemic lupus erythematosus (SLE) and/or diffuse large cell lymphoma (DLBCL) in a genetically/ immunologically susceptible individual with defective cytotoxic T-cell response against EBV. We describe lupus nephritis in a 45 years old male suffering from untreated NHL. CD20+ DLBCL was demonstrated by immunohistochemistry of the neck lymph node (LN) biopsy performed for generalized lymphadenopathy. Renal biopsy revealed class V + IV lupus nephritis. Serology demonstrated EBV infection. Complete clinical remission of both SLE and DLBCL was achieved post-therapy with six-cycle rituximab, cyclophosphamide, vincristin, adriablastin, methylprednisolone (R-CHOP) regime. This case report demonstrated the complex relationships between NHL, SLE, EBV and membranous glomerulonephritis. The presented case is remarkable not only because of the rare association of SLE and DLBCL, but also because of its successful treatment with R-CHOP.


Clinical significance of autoantibody to hepatocyte membrane antigen in type 1 autoimmune hepatitis.
By using HepG2 as flow cytometry target, we have reported that autoantibody to hepatocyte membrane antigen (anti-HMA) was frequently found in autoimmune hepatitis (AIH) patients. In this study, we have examined this autoantibody in relation to clinical features in these patients. HepG2 cells were incubated with diluted serum and subsequently with FITC-conjugated antihuman immunoglobulin. The results were expressed as relative fluorescence intensity. The prevalence of anti-HMA was estimated by setting the upper limit of mean +/- 3 SD obtained from healthy subjects. We found that the mean relative fluorescence intensity was 1.67+/-0.5 in AIH with low serum ALT level (group 1 AIH), 4.20+/-1.9 in AIH with high serum ALT level (group 2 AIH), and 1.92+/-0.9 in age-matched chronic hepatitis C virus-positive patients. Their positive rate was 37.5% (three of eight) in group 1 AIH, 95.0% (19 of 20) in group 2 AIH, and 33.3% (four of 12) in chronic hepatitis C patients. In 12 group 2 AIH patients, their mean relative fluorescence intensity was significantly decreased during immunosuppressive therapy. The association between serum ALT level and anti-HMA was confirmed by the facts that a significant direct quantitative relationship existed between these two levels and by serial studies of anti-HMA in four AIH patients. Anti-HMA was also detected in five non-B, non-C hepatitis patients having clinical features resembling those of AIH. The present results have shown that the anti-HMA was tightly associated with the degree of hepatocyte inflammation and that the measurement of anti-HMA may have some advantage in clinical evaluation of some of non-B, non-C hepatitis patients.


Effect of aminoisobutyric acid (Aib) substitutions on the antimicrobial and cytolytic activities of the frog skin peptide, temporin-1DRa.
Temporin-1DRa (HFLGTLVNLAKKIL.NH(2)), first isolated from the skin of the California red-legged frog Rana draytonii, shows broad-spectrum antimicrobial activity but its therapeutic potential is limited by its toxicity against mammalian cells. The cytolytic properties of cationic alpha-helical peptides are determined by a complex interaction between cationicity, hydrophobicity, conformation, and amphipathicity. This study has investigated the cytolytic properties of conformationally constrained analogs of temporin-1DRa containing alpha-aminoisobutyric acid (Aib) substitutions. Cytolytic activity was determined against the bacteria Escherichia coli and Staphylococcus aureus, the opportunistic yeast pathogen, Candida albicans, human erythrocytes, HepG2 hepatoma-derived cells, and L929 fibroblasts. Aib substitutions at Gly(4), Asn(8), and Ala(10) increased both % helicity, determined in methanol solution, and hydrophobicity resulting in increases in both antimicrobial potencies and toxicities against the mammalian cells. Substitution at Leu(6) resulted in an appreciable decrease in cytolytic activity against all cells whereas the substitutions at His(1), Phe(2), Leu(3), Thr(5), and Val(7) had only minor effects on activity. Substitutions at Leu(9), Ile(13), Leu(14) produced analogs with decreased helicity and hydrophobicity that retained activity against microorganisms but showed appreciably lower cytolytic activities against mammalian cells. In particular, the fourfold increase in therapeutic index [ratio of LC(50) against erythrocytes to minimum inhibitory concentration (MIC) against microorganisms] of [Aib(13)]temporin-1DRa identifies it as a compound with potential for development as a therapeutically valuable anti-infective agent.


An Experimental Investigation of Antisocial Lie-Telling Among Children With Disruptive Behavior Disorders and Typically Developing Children.
Children's lie-telling is surprisingly understudied among children with significant behavioral problems. In the present study, experimental paradigms were used to examine antisocial lie-telling among ethnically diverse 5- to 10-year-old children with disruptive behavior disorders (DBD; n = 71) and a typically developing (TD) comparison sample (n = 50) recruited from a southeastern state from 2013 to 2014. Children completed two games that measured the prevalence and skill of their lies: (a) for personal gain and (b) to conceal wrongdoing. Children with DBD were more likely to lie for personal gain than TD children. With age, children were more likely to lie to conceal wrongdoing, but the reverse was true regarding lies for personal gain. Results advance knowledge concerning individual differences in children's lie-telling.


Risk of Hepatitis B Virus (HBV) Reactivation in Patients with Immune-Mediated Inflammatory Diseases Receiving Biologics: Focus on the Timing of Biologics after Anti-HBV Treatment.
Anti-hepatitis B virus (HBV) therapy is required for patients with HBV infection receiving biologics because of the high risk of HBV reactivation. However, it is unclear when to start biologics after anti-HBV treatment. We investigated the risk of HBV reactivation according to the timing of biologics initiation after anti-HBV treatment in immune-mediated inflammatory disease (IMID) patients with HBV infection. We retrospectively evaluated the incidence of HBV reactivation in IMID patients who received biologics between July 2005 and April 2020. The patients were divided into two groups (within 1-week and after 1-week) according to the timing of biologics initiation after anti-HBV treatment. The cumulative probabilities and factors associated with HBV reactivation were evaluated. A total of 60 hepatitis B surface antigen-positive patients with IMID received biologics (within 1-week group, n=23 [38%]; after 1-week group, n=37 [62%]). During a median follow-up of 34 months (interquartile range, 20 to 74 months), three patients (5%) developed HBV reactivation. In univariate analysis, the timing of biologics after anti-HBV treatment was not significantly associated with the risk of HBV reactivation (hazard ratio, 0.657; 95% confidence interval, 0.059 to 7.327; p=0.733). The cumulative probabilities of HBV reactivation did not significantly differ according to the timing of biologics (p=0.731). The risk of HBV reactivation was not significantly associated with the timing of biologics administration after anti-HBV treatment. Thus, biologics may be initiated early in patients with IMID undergoing treatment for HBV.


Trans-plasma membrane electron transport in mammals: functional significance in health and disease.
Trans-plasma membrane electron transport (t-PMET) has been established since the 1960s, but it has only been subject to more intensive research in the last decade. The discovery and characterization at the molecular level of its novel components has increased our understanding of how t-PMET regulates distinct cellular functions. This review will give an update on t-PMET, with particular emphasis on how its malfunction relates to some diseases, such as cancer, abnormal cell death, cardiovascular diseases, aging, obesity, neurodegenerative diseases, pulmonary fibrosis, asthma, and genetically linked pathologies. Understanding these relationships may provide novel therapeutic approaches for pathologies associated with unbalanced redox state.


Pre-Demosaic Graph-Based Light Field Image Compression.
An unfocused plenoptic light field (LF) camera places an array of microlenses in front of an image sensor in order to separately capture different directional rays arriving at an image pixel. Using a conventional Bayer pattern, data captured at each pixel is a single color component (R, G or B). The sensed data then undergoes demosaicking (interpolation of RGB components per pixel) and conversion to an array of sub-aperture images (SAIs). In this paper, we propose a new LF image coding scheme based on graph lifting transform (GLT), where the acquired sensor data are coded in the original captured form without pre-processing. Specifically, we directly map raw sensed color data to the SAIs, resulting in sparsely distributed color pixels on 2D grids, and perform demosaicking at the receiver after decoding. To exploit spatial correlation among the sparse pixels, we propose a novel intra-prediction scheme, where the prediction kernel is determined according to the local gradient estimated from already coded neighboring pixel blocks. We then connect the pixels by forming a graph, modeling the prediction residuals statistically as a Gaussian Markov Random Field (GMRF). The optimal edge weights are computed via a graph learning method using a set of training SAIs. The residual data is encoded via low-complexity GLT. Experiments show that at high PSNRs-important for archiving and instant storage scenarios-our method outperformed significantly a conventional light field image coding scheme with demosaicking followed by High Efficiency Video Coding (HEVC).


Determination of deoxynivalenol in soft wheat by immunoaffinity column cleanup and LC-UV detection: interlaboratory study.
An interlaboratory study was conducted to assess the applicability of a previously validated method for the analysis of deoxynivalenol (DON) in cereal and cereal products to soft wheat in the range of >0.1-3.0 microg/g. The study evaluated a generic method to determine DON at levels that bracket the existing Canadian guidelines for DON in soft wheat destined for use in baby foods and nonstaple foods. Collaborators selected one of 2 approved brands of DON immunoaffinity column for cleanup and their choice of qualified C18 liquid chromatographic (LC) column. Separation was by LC with UV detection. Blind duplicates from 5 levels of naturally contaminated wheat and a pair of spiked wheat samples were successfully analyzed by 12 laboratories in 8 countries. For samples naturally contaminated with DON from <0.1-2.2 microg/g, the relative standard deviation of repeatability (RSDr) ranged from 3.1 to 14.8%. For reproducibility, the RSDR ranged from 21.0 to 32.9% and the HorRat range was 1.0 to 1.9. Recoveries of 0.5 microg/g DON spiked into wheat ranged from 66 to 98%, with an average of 84%. The RSDr was 5.4%, the RSDR was 12.6%, and the HorRat value was 0.7.


The effect of a perioperative steroid pulse on surgical stress in hepatic resection.
Little has been reported on the control of cytokine release, which is induced by surgical stress. The aim of this study was to investigate the effect of a perioperative short-term steroid pulse on surgical stress. Seventeen patients with a non-cholestatic liver, who underwent hepatic resection, were included in this study. The patients were classified into two groups consisting of a control group (n = 11) and a steroid pulse group (n = 6), in whom 30 mg/kg of methylprednisolone was intravenously administered 3 hours before the operation, and thereafter 1 gram of methylprednisolone was also intravenously administered both immediately after operation and on postoperative day 1. Interleukin 6 was serially measured, and the perioperative clinical parameters including C-reactive protein were compared between both the control and the steroid pulse groups. The IL-6 values immediately after operation, as well as at postoperative days 1 and 3 in the steroid pulse group were significantly lower than those in the control group. The value of the C-reactive protein at postoperative day 3 in the steroid pulse group was significantly lower than that in the control group, and the postoperative peak value of bilirubin as well as the incidence of postoperative complications in the control group tended to be higher than those in the steroid pulse group. In addition, the steroid pulse demonstrated no adverse side effects. A perioperative short-term steroid pulse using methylprednisolone is thus suggested to reduce surgical stress by decreasing cytokine release.


Weakly supervised convolutional LSTM approach for tool tracking in laparoscopic videos.
Real-time surgical tool tracking is a core component of the future intelligent operating room (OR), because it is highly instrumental to analyze and understand the surgical activities. Current methods for surgical tool tracking in videos need to be trained on data in which the spatial positions of the tools are manually annotated. Generating such training data is difficult and time-consuming. Instead, we propose to use solely binary presence annotations to train a tool tracker for laparoscopic videos. The proposed approach is composed of a CNN + Convolutional LSTM (ConvLSTM) neural network trained end to end, but weakly supervised on tool binary presence labels only. We use the ConvLSTM to model the temporal dependencies in the motion of the surgical tools and leverage its spatiotemporal ability to smooth the class peak activations in the localization heat maps (Lh-maps). We build a baseline tracker on top of the CNN model and demonstrate that our approach based on the ConvLSTM outperforms the baseline in tool presence detection, spatial localization, and motion tracking by over [Formula: see text], [Formula: see text], and [Formula: see text], respectively. In this paper, we demonstrate that binary presence labels are sufficient for training a deep learning tracking model using our proposed method. We also show that the ConvLSTM can leverage the spatiotemporal coherence of consecutive image frames across a surgical video to improve tool presence detection, spatial localization, and motion tracking.


Workforce Factors for Psychologists in CAMHS in New Zealand.
Recruitment and retention of psychologists for Child and Adolescent Mental Health Services (CAMHS) is difficult worldwide. This study: 1) identified factors that promote or hinder recruitment and retention of CAMHS psychologists; 2) explored training and its impact on CAMHS as a career choice; and 3) identified strategies to improve recruitment and retention of CAMHS psychologists in New Zealand. Surveys were undertaken with four New Zealand groups: 1) all registered psychologists; 2) final-year professional psychology interns; 3) directors for professional psychology training programmes; and 4) psychology professional leaders in the public health system. Psychologists with more pre-qualification training in CAMHS were more likely to work in CAMHS. Other psychologists reported they would have more likely worked in CAMHS if they had received more pre-qualification training. Factors that promote or hinder training, recruitment and retention are discussed. A range of practical strategies for improving recruitment and retention of psychologists to CAMHS were identified, including particular strategies for psychologists from minority cultures (particularly, Maori and Pacific Island psychologists). While undertaken in New Zealand, many of these findings are relevant internationally.


Microglia regulate chandelier cell axo-axonic synaptogenesis.
SignificanceChandelier cells (ChCs) are a unique type of GABAergic interneuron that form axo-axonic synapses exclusively on the axon initial segment (AIS) of neocortical pyramidal neurons (PyNs), allowing them to exert powerful yet precise control over PyN firing and population output. The importance of proper ChC function is further underscored by the association of ChC connectivity defects with various neurological conditions. Despite this, the cellular mechanisms governing ChC axo-axonic synapse formation remain poorly understood. Here, we identify microglia as key regulators of ChC axonal morphogenesis and AIS synaptogenesis, and show that disease-induced aberrant microglial activation perturbs proper ChC synaptic development/connectivity in the neocortex. In doing so, such findings highlight the therapeutic potential of manipulating microglia to ensure proper brain wiring.


Intravascular Lithotripsy in Calcified Coronary Lesions: A Single-Center Experience in "Real-World" Patients.
This study aims to describe the outcome of intravascular lithotripsy (IVL) when used with different indications and to assess the short- and long-term outcomes of IVL-facilitated percutaneous coronary intervention (PCI). Intravascular lithotripsy can improve the results of PCI of calcified coronary lesions with a low rate of periprocedural complications. A total of 105 consecutive patients with 110 calcified lesions underwent IVL. A total of 87 de novo lesions were treated by IVL with the following indications: 25 before attempting other balloon-based devices (primary IVL), 51 after the failure of non-compliant balloon dilatation (secondary IVL), and 11 after stent implantation because of stent under expansion (bailout IVL). In 23 lesions, IVL was used for the treatment of in-stent restenosis (ISR). Effectiveness (angiographic success) and safety [major adverse cardiovascular events (MACEs) and IVL-related procedural complications] endpoints were assessed. Angiographic success was achieved in 84.6% of lesions. Early MACEs were periprocedural MI only, ranging from 6.7 to 20% depending on MI definition. The flow-limiting dissections rate was 2.7%. A total of five (4.5%) IVL balloons ruptured during treatment with subsequent vessel perforation in 1 case. MACEs at 12 months were 13.3%, with TLR occurring in 8 lesions (12% primary IVL, 0% secondary IVL, 0% bailout IVL, and 21.7% IVL for ISR, p = 0.002). Treatment of calcified coronary lesions with IVL in a "real-world" setting can be performed with high success, low rate of procedural complications, and an acceptable MACEs rate. Target lesion failure may be more frequent when IVL is performed for the treatment of ISR due to calcium-mediated stent under expansion.


T-cell selection in the thymus: a spatial and temporal perspective.
The ability of T cells to respond to a wide array of foreign antigens while avoiding reactivity to self is largely determined by cellular selection of developing T cells in the thymus. While a great deal is known about the cell types and molecules involved in T-cell selection in the thymus, our understanding of the spatial and temporal aspects of this process remain relatively poorly understood. Thymocytes are highly motile within the thymus and travel between specialized microenvironments at different phases of their development while interacting with distinct sets of self-peptides and peptide presenting cells. A knowledge of when, where, and how thymocytes encounter self-peptide MHC ligands at different stages of thymic development is key to understanding T-cell selection. In the past several years, our laboratory has investigated this topic using two-photon time-lapse microscopy to directly visualize thymocyte migration and signaling events, together with a living thymic slice preparation to provide a synchronized experimental model of T-cell selection in situ. Here, we discuss recent advances in our understanding of the temporal and spatial aspects of T-cell selection, highlighting our own work, and placing them in the context of work from other groups.


Eosinophil locomotion and the release of IL-3 and IL-5 by allergen-stimulated mononuclear cells are effectively downregulated in vitro by budesonide.
Treatment of allergic asthma with inhaled corticosteroids, such as budesonide (BDN), results in downregulation of T-cell activation and of eosinophil recruitment. Since blood concentrations of BDN, although significantly lower than those measured in the lung, may still have anti-inflammatory effects, we evaluated the activity of BDN in vitro on: allergen-induced release of lymphokines involved in eosinophil chemotaxis (i.e. IL-3 and IL-5), at drug concentrations similar to those obtained in vivo in the lung (10-8 M), and eosinophil locomotion, at "systemic concentrations' of the drug (10(-10)M and 10(-9)M). Twenty-three atopic asthmatic subjects (atopics) sensitized to Dermatophagoides pteronyssinus (Dp) and seven non-atopic healthy subjects (controls) were studied. Purified blood mononuclear cells (BMC) were stimulated with Dp, with or without BDN 10(-8) M and, after 6 days, the supernatants were collected and frozen to test their chemotactic activity toward purified blood eosinophils and their levels of interleukin (IL)-3 and IL-5 by immunoassay. BMC were then pulsed for additional 18 h with [3H]thymidine to evaluate allergen-induced T-cell proliferation. In addition, to test possible direct effects of 'systemic concentrations' of the drug on eosinophil locomotion, blood eosinophils were incubated for 1 h with BDN (10(-10) M and 10(-9) M) prior to test their chemotactic response toward recombinant human IL-3 and IL-5. Stimulation of BMC from atopics with Dp induced a statistically significant increase in [3H]thymidine incorporation (P < 0.05); secretion of chemotactic factors for eosinophils (P < 0.001) and the release of IL-3 and IL-5 (P < 0.005 and P < 0.05 respectively). BDN, at the concentration of 10(-8) M, was able to significantly down-regulate T-cell proliferation (P < 0.05), the secretion of chemotactic factors for eosinophils (P < 0.001) and the release of IL-3 and IL-5 (P < 0.01 and P < 0.05 respectively). Similarly, "systemic concentrations' of BDN (10(-10) M and 10(-9) M) totally inhibited the chemotactic response of blood eosinophils toward recombinant human IL-3 and IL-5 (P < 0.005). Concentrations of BDN similar to those obtained in vivo are effective in inhibiting both the release of eosinophils chemotaxins by allergen-activated mononuclear cells and eosinophil locomotion.


Exploratory study on the psychological impact of COVID-19 on the general Brazilian population.
The COVID-19 pandemic has become one of the main international concerns regarding its impact on mental health. The present study aims to investigate the prevalence of depression, anxiety, and stress symptoms, and behavioral aspects amidst the COVID-19 pandemic in a Brazilian population. An online survey was administered from May 22 to June 5, 2020 using a questionnaire comprising of sociodemographic information, the Depression, Anxiety, and Stress Scale (DASS-21), and the Coping Strategies Inventory. Participants comprised 3,000 people from Brazil's 26 states and the Federal District, with an average age of 39.8 years, women (83%), married (50.6%), graduates (70.1%) and employees (46.7%). Some contracted the virus (6.4%) and had dead friends or relatives (22.7%). There was more consumption of drugs, tobacco, medication, and food (40.8%). Almost half of participants expressed symptoms of depression (46.4%), anxiety (39.7%), and stress (42.2%). These were higher in women, people without children, students, patients with chronic diseases, and people who had contact with others diagnosed with COVID-19. The existence of a group more vulnerable to situations with a high stress burden requires greater attention regarding mental health during and after the pandemic. That said, it should be emphasized that these findings are preliminary and portray a moment still being faced by many people amid the pandemic and quarantine measures. Therefore, we understand that the magnitude of the impacts on mental health will only be more specific with continuous studies after total relaxation of the quarantine.


Spatiotemporal monitoring endocytic and cytosolic pH gradients with endosomal escaping pH-responsive micellar nanocarriers.
Endosomal escape is of crucial importance to increase the therapeutic efficacy for nanoparticle-based drug and gene delivery. It has been long presumed that pH-responsive polymeric nanocarriers are potent in aiding endosomal escape due to the "proton sponge" effect; however, the intracellular pH (pHi) gradients subjected by pH-responsive nanocarriers during endocytic and endosomal escaping processes remain to be quantified and elucidated. We herein report the fabrication of ultrasensitive ratiometric fluorescent pHi imaging probes with robust endosomal escaping capability derived from dual dye-labeled pH-responsive block copolymers, which can directly monitor endosomal escape in living cells and quantitatively measure pHi variations during the entire endocytic and endosomolytic processes. Micellar nanoparticle-based pHi sensors could be efficiently internalized into cells via endocytosis where micelle-to-unimer transition occurs, followed by endosomal escape into the cytosol. This process is accompanied by deactivation of blue coumarin emission within acidic organelles and restored blue/red dual emissions within the neutral cytosolic milieu, allowing for ratiometric fluorescent imaging of entire pHi gradients subjected by micellar nanoparticles following the endocytic transport pathway.


[Level of enkephalins and cyclic nucleotides in the myocardium in ventricular fibrillation caused by experimental myocardial infarct].
Ventricular fibrillation, caused by the occlusion of the anterior descending coronary artery, results in leu-enkephalin level decrease in ischemic and non-ischemic zones of myocardium. The fibrillation is found to induce cAMP increase in the ischemic zone and cGMP increase in the normal circulation zone. cGMP and leu-enkephalin level correlates with fibrillation duration. Enkephalins and cyclic nucleotides are suggestive of being of importance in heart rythm disturbances pathogenesis.


Usefulness of ultrasonography in the treatment of nasal bone fractures.
Ultrasonography has recently been used in assessment and diagnosis of maxillofacial trauma because it is easy and quick to perform, inexpensive, portable, and noninvasive. The aim of this study was to estimate ultrasonography as an intraoperative repositioning control of nasal bone fractures. We studied 32 patients with suspected nasal bone fracture. We performed preoperative computed tomography (CT) for evaluation of the type and extent of nasal bone fractures. We also took external photographs for evaluation of external deviation of the nose and nasal deformity. During surgery, we performed real-time ultrasonography-guided closed reduction using a 10 MHz linear transducer. After 1 year, we performed postoperative evaluation with CT and external photography. We classified patients into three groups according to their CT score. Patients were 23 males and 9 females aged 8-39 years. Clinical symptoms were pain, nasal swelling, nasal bleeding, and localized depression at the trauma site. In almost all patients, postoperative external photographs showed a symmetrical nasal dorsum without external deformity, and postoperative CT showed stabilization of bony fragments and good alignment of the nasal bone. Postoperatively, the CT score was 3 (excellent) in 25 patients, 2 (good) in 5 patients, and 1 (fair) in 2 patients. We suggest that ultrasonography is very useful for evaluating intraoperative repositioning of nasal bone fractures.


Malignant melanomas in the small intestine: a study of 103 patients.
Malignant melanoma shows an unusual predilection to metastasize to the small intestine. A proportion of small bowel melanomas occur without history of an antecedent primary. We evaluated a group of patients with malignant melanoma in the small intestine to further our understanding of this disease. We reviewed 103 cases of malignant melanoma in the small intestine (77 surgical resections and 26 autopsies) accessioned at the Armed Forces Institute of Pathology between 1945 and 1991 for demographic, chronological, and pathological features. Mean age at time of primary was 45.6 yr for surgical and 34.1 yr for autopsy cases (p = 0.01). Mean age at time of small intestinal involvement was 52.2 yr for surgical and 42.7 yr for autopsies (p = 0.03). Primary lesions preceded intestinal disease by an average of 5.6 yr for surgical and 2.1 yr for autopsies. The age distribution of surgical patients with and without known primary melanomas at the time of small intestinal melanoma was not significantly different. The same was true for autopsy patients. Using regression analysis, the linear relationship of age at primary melanoma (AAP) on age at small intestinal melanoma (AASI) was given by AAP = 2.30 + 0.85 (AASI), and that of AASI on AAP was given by AASI = 3.94 + 1.02 (AAP) (r = 0.93 and p < 0.0001 for both regressions). Our data and results support the concept that small bowel involvement by melanoma, even without a known primary, is most probably metastatic. The age at which an unknown primary occurred in cases of intestinal melanoma, or the age at which intestinal metastasis may appear in cases with known primary melanoma, can be estimated. There appear to be two subsets of primary melanoma: one that occurs among younger patients and is more aggressive with rapid metastasis and early death and one that occurs among older patients, is more indolent, and metastasizes less rapidly.


Adenosine dysfunction and adenosine kinase in epileptogenesis.
Traditionally, epilepsy has been considered to be a disorder of neuronal dysfunction. Based on this dogma, drug development efforts have largely focused on neurocentric model systems to screen for compounds that affect the function of neurons. Unfortunately, about 30% of all patients with epilepsy - or more than 20 million worldwide - are refractory to classical neurocentric pharmacotherapy. The failure of neurocentric pharmacotherapy in epilepsy requires radical rethinking and the search for novel therapeutic targets. Research from recent years suggests that epilepsy is a disorder of astrocyte dysfunction. Astrocytes are key regulators of the brain's own anticonvulsant adenosine. Thus, any dysfunction in astrocyte metabolism will drastically affect the brain's ability to control excitability via adenosinergic neuromodulation. This review will focus on the astrocyte-based enzyme adenosine kinase (ADK) as the key regulator of synaptic adenosine. Astrogliosis - a pathological hallmark of the epileptic brain - leads to overexpression of the adenosine-removing enzyme ADK and therefore to adenosine deficiency. Evidence from transgenic animals demonstrates that overexpression of ADK per se is sufficient to trigger seizures. Consequently, pharmacological inhibition of ADK is very effective in suppressing seizures that are refractory to classical antiepileptic drugs. The recent definition of ADK as rational target to predict and to prevent seizures in epilepsy has prompted the development of focal adenosine augmentation therapies (AATs) that have been designed to selectively reconstitute adenosinergic signalling within an area of astrogliosis-based adenosine-dysfunction. This therapeutic challenge has experimentally been met with polymeric or stem cell based brain implants to afford the focal delivery of adenosine.


A novel mutation c.1048A>T at codon 350(Lys>Stop) in PROC gene causing neonatal purpura fulminans.
Purpura fulminans in the neonatal period due to severe congenital protein C deficiency (protein C activity <1 IU/dl) is a rare autosomal recessive disorder. If untreated, it is fatal. Early identification of such patients may be lifesaving. Acquired deficiency of protein C caused by increased consumption as overt disseminated intravascular coagulation (DIC) and severe infection creates a diagnostic dilemma. Mutation analysis plays a critical role in confirming the diagnosis of the disease and offering prenatal diagnosis. In this report, we describe a newborn who presented with purpura fulminans and DIC, molecular analysis showed a novel c.1048A>T transversion in a homozygous state at codon 350 (Lys>Stop) of protein C (PROC) gene. Prenatal diagnosis in subsequent pregnancy was done which revealed the affected fetus had the same mutation in homozygous form.


Neurological Alterations in Type 1 Diabetes Mellitus Among Adolescents.
Diabetes mellitus (DM) is a group of chronic disorders of metabolism characterised by high blood glucose levels. There is an increased prevalence of Type 1 DM in children and adolescents with its adverse complications especially microvascular ones (retinopathy, nephropathy and neuropathy) that might cause multiple organ damage. To study the relation between DM and neurological affection. Fifty-nine children with type I DM, divided randomly into 2 groups, aged 8-18 years old of both sexes were enrolled in this cross-sectional study. All children were subjected to full history taking, physical, neurological and systemic examination. There was an affection of motor power in both upper limbs as well as lower limbs. Also, we found that there was an affection of the superficial peripheral sensation affecting both upper and lower limbs. Neurological assessment of children with diabetes mellitus type I should be a routine to early discover these manifestations which can have a deteriorating effect on the child's health.


Electric-field-induced fluorescence quenching in polyfluorene, ladder-type polymers, and MEH-PPV: evidence for field effects on internal conversion rates in the low concentration limit.
Electric field-induced fluorescence quenching has been measured for a series of conjugated polymers with applications in organic light-emitting diodes. Electrofluorescence measurements on isolated chains in a glassy matrix at 77 K show that the quenching efficiency for poly[2-methoxy-5-(2-ethylhexyloxy)-p-phenylenevinylene] (MEH-PPV) is an order of magnitude larger than that for either a ladder-type polymer (MeLPPP) or polyfluorene (PFH). This effect is explained in terms of the relatively high probability of field-enhanced internal conversion deactivation in MEH-PPV relative to either MeLPPP or PFH. These data, obtained under dilute sample conditions such that chain-chain interactions are minimal, are contrasted with the much higher quenching efficiencies observed in the corresponding polymer films, and several explanations for the differences are considered. In addition, the values of the change in dipole moment and change in polarizability on excitation (|Δμ| and tr(Δα), respectively) are reported, and trends in these values as a function of molecular structure and chain length are discussed.


Impaired temporal contrast sensitivity in dyslexics is specific to retain-and-compare paradigms.
Developmental dyslexia is a specific reading disability that affects 5-10% of the population. Recent studies have suggested that dyslexics may experience a deficit in the visual magnocellular pathway. The most extensively studied prediction deriving from this hypothesis is impaired contrast sensitivity to transient, low-luminance stimuli at low spatial frequencies. However, the findings are inconsistent across studies and even seemingly contradictory. In the present study, we administered several different paradigms for assessing temporal contrast sensitivity, and found both impaired and normal contrast sensitivity within the same group of dyslexic participants. Under sequential presentation, in a temporal forced choice paradigm, dyslexics showed impaired sensitivity to both drifting and flickering gratings. However, under simultaneous presentation, with a spatial forced choice paradigm, dyslexics' sensitivity did not differ from that of the controls. Within each paradigm, dyslexics' sensitivity was poorer at higher temporal frequencies, consistent with the magnocellular hypothesis. These results suggest that a basic perceptual impairment in dyslexics may be their limited ability to retain-and-compare perceptual traces across brief intervals.


Identification of pp120, an endogenous substrate for the hepatocyte insulin receptor tyrosine kinase, as an integral membrane glycoprotein of the bile canalicular domain.
An endogenous membrane-bound substrate of the insulin receptor beta-subunit tyrosine kinase in liver, pp120, has been identified as HA4, a 110-kDa membrane glycoprotein localized primarily to the bile canalicular domain of the hepatocyte. HA4 has been implicated in bile salt transport and cell adhesion. Monoclonal antibodies to HA4 were used to identify it as a substrate of the insulin receptor kinase. Anti-pp120 and anti-HA4 were found to cross-react, and phosphopeptide maps for each of the corresponding antigens were identical. The identification of pp120 as HA4 serves to link insulin action through the receptor tyrosine kinase activity to bile metabolism and raises questions pertaining to the intracellular site(s) of action of the insulin receptor.


[Two sisters with progressive familial intrahepatic cholestasis].
A few months after birth two sisters aged 5 and 9 years had developed cholestasis and abnormal liver functions with symptoms including itching and jaundice. The younger sister also developed rickets and clotting disorders. On clinical, biochemical and immunohistopathological grounds the diagnosis of 'progressive familial intrahepatic cholestasis (PFIC) type 2' was made. Medical treatment was not succesfull in reducing symptoms. An ileocolonic bypass in the younger child was not effective. Subsequently, both patients underwent partial external biliary diversion (PEBD). Except for a period of intermittent itching in the younger child, both patients remained free of symptoms 2 years after PEBD. In cases where an early diagnosis is made, PEBD could delay or even prevent the necessity of liver transplantation.


Development of a neutron-activated concrete powder reference material.
In this paper, the development of a neutron-activated concrete powder reference material is described. The material originated from core samples taken from a concrete bioshield of a decommissioned nuclear reactor which ceased operation almost 30 years ago after approximately 20 years of operation. The assigned values, which were in the Bq g(-1) range, for the radionuclides in the material were determined by a 'consensus' method from measurements made by 33 organisations from 15 countries. The measurements were made within a wider test exercise (the NPL Environmental Radioactivity Proficiency Test Exercise 2008). Assigned specific activity values were obtained for (60)Co, (133)Ba, (152)Eu and (154)Eu and indicative values were obtained for (3)H (total), (3)H (leachable), (14)C, (40)K, (55)Fe and (63)Ni.


Enzyme alchemy: cell-free synthetic biochemistry for natural products.
Cell-free synthetic biochemistry aims to engineer chemical biology by exploiting biosynthetic dexterity outside of the constraints of a living cell. One particular use is for making natural products, where cell-free systems have initially demonstrated feasibility in the biosynthesis of a range of complex natural products classes. This has shown key advantages over total synthesis, such as increased yield, enhanced regioselectivity, use of reduced temperatures and less reaction steps. Uniquely, cell-free synthetic biochemistry represents a new area that seeks to advance upon these efforts and is particularly useful for defining novel synthetic pathways to replace natural routes and optimising the production of complex natural product targets from low-cost precursors. Key challenges and opportunities will include finding solutions to scaled-up cell-free biosynthesis, as well as the targeting of high value and toxic natural products that remain challenging to make either through whole-cell biotransformation platforms or total synthesis routes. Although underexplored, cell-free synthetic biochemistry could also be used to develop 'non-natural' natural products or so-called xenobiotics for novel antibiotics and drugs, which can be difficult to engineer directly within a living cell.


An energy-barrier model for the permeation of monovalent and divalent cations through the maxi cation channel in the plasma membrane of rye roots.
The depolarization-activated, high-conductance "maxi" cation channel in the plasma membrane of rye (Secale cereale L.) roots is permeable to a wide variety of monovalent and divalent cations. The permeation of K+, Na+, Ca2+ and Ba2+ through the pore could be simulated using a model composed of three energy barriers and two ion binding sites (a 3B2S model), which assumed single-file permeation and the possibility of double cation occupancy. The model had an asymmetrical free energy profile. Differences in permeation between cations were attributed primarily to differences in their free energy profiles in the regions of the pore adjacent to the extracellular solution. In particular, the height of the central free energy peak differed between cations, and cations differed in their affinities for ion binding sites. Significant ion repulsion occurred within the pore, and the mouths of the pore had considerable surface charge. The model adequately described the diverse current vs. voltage (I/V) relationships obtained over a wide variety of experimental conditions. It described the phenomena of non-Michaelian unitary conductance vs. activity relationships for K+, Na+ and Ca2+, differences in selectivity sequences obtained from measurements of conductance and permeability ratios, changes in relative cation permeabilities with solution composition, and the complex effects of Ba2+ and Ca2+ on K+ currents through the channel. The model enabled the prediction of unitary currents and ion fluxes through the maxi cation channel under physiological conditions. It could be used, in combination with data on the kinetics of the channel, as input to electrocoupling models allowing the relationships between membrane voltage, Ca2+ influx and Ca2+ signaling to be studied theoretically.


SIRT1 Interacts with and Deacetylates ATP6V1B2 in Mature Adipocytes.
SIRT1 plays a key role in maintaining metabolic homeostasis in mammals by directly modulating the activities of various transcription factors and metabolic enzymes through lysine deacetylation. White adipose tissue plays a key role in lipid storage and metabolism. To identify novel molecular targets of SIRT1 in fat cells, we used a non-biased proteomic approach. We identified a number of proteins whose acetylation status was significantly affected by SIRT1 modulator treatment in 3T3-L1 adipocytes. Among them, ATP6V1B2, a subunit of the vacuolar (H+)-ATPase, was further shown to be associated with SIRT1 by co-immunoprecipitation assay. Moreover, SIRT1 deacetylates ATP6V1B2 in vitro and in vivo. Taken together, our study demonstrates that ATP6V1B2 is a molecular target of SIRT1 in fat cells and the role of SIRT1 and ATP6V1B2 acetylation in the vacuolar (H+)-ATPase function warrants further investigation.


Pushing the Envelope in Psoriasis: Late Cornified Envelope Proteins Possess Antimicrobial Activity.
Deletion of late cornified envelope (LCE) genes LCE3B and LCE3C (LCE3B/C-del) is a psoriasis risk factor linked to the major psoriasis risk gene HLA-C*06. Niehues et al. demonstrate that LCE3B/C-del leads to increased keratinocyte LCE3A expression. They also show that LCE3A/B/C possess antimicrobial activity but do not obviously regulate epidermal barrier integrity. These findings implicate LCE proteins in psoriasis pathogenesis via a new functional role.


Update on the diagnosis and management of gout.
Gout is a common clinical problem encountered by both general and specialist clinicians. The key principles in gout management include establishing a definitive diagnosis, the swift treatment of acute attacks, and using urate-lowering therapies appropriately to prevent further attacks and joint damage. The gold standard diagnostic tool for gout remains the identification by polarised light microscopy of monosodium urate crystals in synovial fluid or in a tophus. Emerging diagnostic imaging techniques and novel therapies show promise in the diagnosis and treatment of gout. In most cases, using existing therapies judiciously remains the key determinant of success in managing gout.


A Novel Delivery System for the Controlled Release~of Antimicrobial Peptides: Citropin 1.1 and Temporin A.
Antimicrobial peptides (AMPs) are prospective therapeutic options for treating multiple-strain infections. However, clinical and commercial development of AMPs has some limitations due to their limited stability, low bioavailability, and potential hemotoxicity. The purpose of this study was to develop new polymeric carriers as highly controlled release devices for amphibian peptides citropin 1.1 (CIT) and temporin A (TEMP). The release rate of the active pharmaceutical ingredients (APIs) was strongly dependent on the API characteristics and the matrix microstructure. In the current work, we investigated the effect of the polymer microstructure on in vitro release kinetics of AMPs. Non-contact laser profilometry, scanning electron microscopy (SEM), and differential scanning calorimetry (DSC) were used to determine the structural changes during matrix degradation. Moreover, geno- and cytotoxicity of the synthesized new carriers were evaluated. The in vitro release study of AMPs from the obtained non-toxic matrices shows that peptides were released with near-zero-order kinetics. The peptide "burst release" effect was not observed. New devices have reached the therapeutic concentration of AMPs within 24 h and maintained it for 28 days. Hence, our results suggest that these polymeric devices could be potentially used as therapeutic options for the treatment of local infections.


Interactions of ouabain and noradrenaline in isolated rabbit's atria.
1. The present study was made using sympathetic nerve-atria preparations, conventional atrial preparations and left atria isolated from rabbits in order that interactions of ouabain and noradrenaline on the transmembrane potential of S-A nodal pacemaker fibres, the pacemaker rate and the contractile force of the left atrium could be investigated.2. In pacemaker action potentials, the maximal diastolic potential was decreased, the early repolarization was accelerated and the late repolarization was slowed by toxic concentrations of ouabain. Oscillating potentials of the diastolic pacemaker membrane were produced.3. Stimulation of the postganglionic sympathetic nerve caused an increase in the slope of diastolic depolarization, resulting in an acceleration of the pacemaker rate. In the presence of ouabain, the pacemaker action potential configuration was significantly altered by nerve stimulation in preparations which showed ouabain-induced bradycardia, but the potential configuration was not changed by nerve stimulation in preparations which showed ouabain-induced tachycardia. Cardiac noradrenaline altered the ion permeability of the cardiac pacemaker membrane during diastole, but it had no effect during systole. Sympathetic nerve stimulation transiently corrected the overshoot and the repolarization of atrial action potentials altered by toxic concentrations of ouabain.4. The positive chronotropic effect of sympathetic nerve stimulation was not affected by either therapeutic or toxic concentrations of ouabain. Ouabain shifted the concentration-chronotropic response curve for exogenous noradrenaline to the left, but did not shift the curve for tyramine.5. Relationship between the developed tension and the rate of contraction was altered differently by ouabain and by noradrenaline. The positive inotropic effect of noradrenaline was not significantly altered by therapeutic concentrations of ouabain but it was inhibited by toxic concentrations.


Efficacy and pattern of use of sputum cytology as a diagnostic test.
Sputum cytology is regarded by many clinicians as a noninvasive, cheap and simple test for the diagnosis of bronchogenic carcinoma. Since the introduction of fibre-optic bronchoscopy and more easily obtained bronchial biopsies reliance on sputum cytology has diminished. However, in Edinburgh it was perceived that sputum samples were still being sent as well as, rather than instead of, bronchoscopic specimens. This retrospective study was undertaken to determine whether or not cytological examination of sputum is an efficient and sensitive test in the investigation of patients with suspected bronchogenic carcinoma. It demonstrated that the Lothian University Hospitals NHS Trust Pathology Directorate receives many sputa from departments not specializing in respiratory disease when there is no indication for the test. In addition, we have shown that the absolute sensitivity of the test is only 5% and that when there is a strong clinical suspicion of bronchogenic carcinoma the results of sputum cytology do not play a significant role in the management of the patient. We recommend that sputum cytology is restricted to those patients under the care of Respiratory Units in whom bronchoscopy is inappropriate or unsuccessful.


Longitudinal stability of serum immunoglobulin G responses to periodontal bacteria.
The value of seroepidemiology in the study of periodontal infections has not been adequately explored. This study examined serum immunoglobulin (IgG) responses to periodontal bacteria in patients with periodontitis and periodontitis-free individuals over a 30-month period. Eighty-nine patients with chronic periodontitis and 42 control subjects with no deep periodontal pockets and no or minimal attachment loss (30-72 years old, 43% men) were included. Patients were examined at baseline, after completed periodontal therapy 4 months post-baseline, and at 30 months, and controls, at baseline and 30 months. IgG antibodies to 19 periodontal species were determined by checkerboard immunoblotting. On average, patients displayed at baseline up to 800-fold higher titers than controls to all but three species. Over the 30-month period, titers remained stable at low levels in controls. In patients, periodontal conditions improved from a baseline mean probing depth of 3.6 mm, bleeding on probing of 62% and an average of 21.5 pockets of=6 mm/person, to 2.5 mm mean pocket depth, 30% bleeding on probing, and 1.2 deep pockets, at 30 months. Over time, antibody titers showed a modest decline in patients, but remained significantly elevated at 30 months in comparison with controls. Antibody-level changes over time were not significantly different between subjects that did and did not receive adjunctive systemic antibiotics. Conspicuous differences in IgG titers to periodontal bacteria exist between periodontitis patients and periodontally healthy controls. Despite successful periodontal therapy, titers remained elevated over a 30-month period, suggesting that serology may mark the history of past periodontal infection.


Bronchoalveolar lavage in radiation pneumonitis.
Bronchoalveolar lavage (BAL) was carried out in six patients with radiation pneumonitis, the early radiation-induced lung damage that usually leads to radiation fibrosis. Protein analysis by immuno-electrophoresis and polyacrylamide gel electrophoresis of BAL fluid revealed leakage of circulatory proteins, including high molecular weight components. Cell count of BAL fluid showed an increased number of lymphocytes; these proved to be activated on cell cycle analysis in one patient. Collagenolytic activity, assessed by degradation of radiolabelled type I human collagen, was present in BAL fluid of all six patients. The following mechanisms are therefore considered to participate in the pathogenesis of radiation-induced lung damage: 1) permeability oedema, which led to acute respiratory distress syndrome in one case of hyperacute radiation pneumonitis, 2) lymphocyte alveolitis possibly perpetuated by activated lymphocytes, and 3) release of collagenolytic enzymes in alveolar structures contributing to fibrotic processes.


Morphology, ultrastructure, and probable functions of the sense organs on the ovipositor stylets of the hymenoptran parasitoid, Venturia canescens (Gravenhorst).
To help understand the mechanism of host discrimination during oviposition of a parasitoid wasp, Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae), the ultrastructure and morphology of the sense organs on the dorsal and ventral stylets of the ovipositor were examined using scanning and transmission electron microscopy. Ovipositor stylets were scanned for sense organs, because only unsheathed ovipositors are used for probing, stabbing, and host discrimination. Five types of sensilla and secretary pores were identified. Based on their pattern of innervation, the surface-dome and pitted-dome sensilla were thought to have a gustatory function and, thus, are the best candidates for host-discrimination. The arrangements of dendrites of type I and type II campaniform sensilla suggested that these acted as pressure- and mechano-receptors, respectively. Coeloconic sensilla might act as thermo-hygroreceptors, whilst secretary pores might provide lubrication during the process of stabbing for oviposition. Surface-dome sensilla were the most numerous type found followed by secretary pores, pitted-dome sensilla, and coeloconic sensilla. Almost all the sensilla and secretary pores were more numerous on the dorsal than on the ventral stylets, and the maximum number of these sensilla and secretary pores occurred on the distal end of the ovipositor stylets and decreased toward the proximal end. The surface-dome sensilla and secretary pores occurred in alternate rows while the pitted-dome sensilla were scattered all over the surface of the ovipositor stylets, especially on the sides of the stylets.


HER2/Neu and the Ets transcription activator PEA3 are coordinately upregulated in human breast cancer.
HER2/Neu is overexpressed in 25-30% of all human breast cancers as a result of both gene amplification and enhanced transcription. Transcriptional upregulation of HER2/neu leads to a 6-8-fold increased abundance of its mRNA per gene copy and likely results from the elevated activity of transcription factors acting on the HER2/neu promoter. Here we report that transcripts of PEA3, an ETS transcription factor implicated in oncogenesis, were increased in 93% of HER2/Neu-overexpressing human breast tumor samples. Analyses to uncover the molecular basis for elevated PEA3 transcripts in HER2/Neu-positive breast tumors revealed that the HER2/Neu receptor tyrosine kinase initiated an intracellular signaling cascade resulting in increased PEA3 transcriptional activity; transcriptionally-activated PEA3 stimulated HER2/neu and PEA3 gene transcription by binding to sites in the promoters of these genes. PEA3 also activates transcription of genes encoding matrix-degrading proteinases, enzymes required for tumor cell migration and invasion. These findings implicate PEA3 in the initiation and progression of HER2/Neu positive breast cancer, and suggest that PEA3 and signaling proteins affecting its regulation are appropriate therapeutic targets.


Breathing pauses during sleep: can a non-invasive ENT examination help identify subjects at risk in epidemiological settings?
In patients with obstructive sleep apnoea (OSA) anatomic and functional upper airway abnormalities are frequent and severe. Invasive methods are used to identify and quantitate the obstruction, to precisely locate its site, etc. as part of pre-treatment or of preoperative evaluations.These methods (lateral skull radiographs, computerized tomography, MRI, fibroscopies, etc) are too expensive and too invasive to be utilized in field surveys. To the classical sleep questionnaires and anthropometric measurements, some simple nose-throat examinations, easily accepted by the volunteers in a population study, could add useful information for the identification of the subjects at risk for sleep-disordered breathing. The present paper is a review of these examinations and of their utility.


[Rupture of type III acute aortic dissection into the bilateral pleural space: report of a case].
Rupture of type III aortic dissection into the bilateral pleural space is very rare. A 62-year-old man was transferred to our department because of aortic dissection for emergency operation. computed tomography (CT) scan revealed rupture of type III aortic dissection into the bilateral pleural space. He underwent successful prosthetic graft replacement through left thoracotomy and removal of a massive amount of blood from the right pleural space by chest tube drainage. The postoperative course was uneventful after 2 days of respiratory support.


The federal government's use of Title VI and Medicare to racially integrate hospitals in the United States, 1963 through 1967.
Explicit discrimination against minorities existed in the 1960s in hospital patient admissions and physician and nurse staff appointments. With passage of the Civil Rights Act of 1964, along with Medicare legislation in 1965, civil rights advocates within the federal government had both a legislative mandate to guarantee equal access to programs funded by the federal government in Title VI and a federal program that affected every hospital in the country in Medicare. This study was conducted to determine the extent to which the Medicare hospital certification program was a major determinant in the racial integration of hospitals throughout the United States. In-depth interviews were conducted with individuals involved in hospital and health care policy in the 1950s and 1960s. Other primary resources include archival and personal manuscripts, government documents, newspapers, and periodicals.


Chemical redox reactions induced by cryptoelectrons on a PMMA surface.
We show that pristine PMMA can spontaneously transfer electrons to species in a liquid, thereby inducing a variety of electron transfer reactions. The electrons that are transferred we call cryptoelectrons; these have a surface density of the order of 5 x 10(13) cm(-2) and are at a considerably more negative reduction potential than the PMMA bonding electrons. For example, metal ions including Ag(+), Cu(2+), and Pd(2+) were reduced and plated on a PMMA surface and Fe(CN)(6)(3-) was reduced to Fe(CN)(6)(4-). Moreover, protons were reduced when PMMA powder was dropped into a slightly acidic solution, resulting in a pH increase and hydrogen generation. Chemiluminescence was produced in a solution containing Ru(bpy)(3)(2+) and S(2)O(8)(2-) with the addition of PMMA powder. These results clearly demonstrate that there are available electrons in PMMA that can participate in redox reactions at a rather negative potential. We also show that contacting PMMA with Teflon depletes this electronic surface charge. However, the PMMA used for a redox reaction or contacted with Teflon that was depleted of the electronic surface charge could be recharged by contacting with a suitable reductant.


Multidisciplinary therapy including high-dose chemotherapy followed by peripheral blood stem cell transplantation for invasive thymoma.
We describe two patients with invasive thymomas who responded to high-dose chemotherapy followed by peripheral blood stem cell transplantation (PBSCT) combined with surgery and radiotherapy. The first patient was a 42-year-old man admitted to the hospital with chest pain, and the second patient was a 45-year-old man admitted with myasthenia gravis. Both patients had nonresectable thymomas (stage IVa) because of invasion of the aorta, pulmonary artery, or both, and dissemination to the pericardium. They initially received two cycles of chemotherapy consisting of adriamycin (40 mg/m(2), day 1), cisplatin (50 mg/m(2), day 1), vincristine (0.6 mg/m(2), day 3), and cyclophosphamide (700 mg/m(2), day 4) at 3-week intervals. Four weeks later, they were administered high-dose etoposide (300 mg/m(2), days 1 to 5) followed by granulocyte colony-stimulating factor (G-CSF) [50 micro g/m(2)/d] subcutaneously to mobilize stem cells into the blood. After two additional cycles of adriamycin, cisplatin, vincristine, and cyclophosphamide (ADOC), the patients received high-dose ifosfamide (1.5 g/m(2), days 1 to 4), carboplatin (400 mg/m(2), days 3 to 5), and etoposide (200 mg/m(2), days 1 to 5) followed by PBSCT. They were administered G-CSF (50 micro g/m(2)/d) after PBSCT, with subsequent rapid recovery of neutrophil and platelet level. The tumors shrank remarkably, and could be excised completely in both patients. Postoperatively, 50 Gy of irradiation was administered. Disease-free status has been maintained for 5 years in the first patient and 2 years in the second patient. Our findings suggest that high-dose ifosfamide, carboplatin, and etoposide followed by PBSCT in combination with an ADOC regimen, surgery, and radiotherapy is very effective and well tolerated in patients with advanced nonresectable thymoma.


Effects of exercise training on the biochemical pathways associated with sarcopenia.
Sarcopenia is considered one of the major causes of disability in the elderly population and is highly associated with aging. Exercise is an essential strategy for improving muscle health while aging and involves multiple metabolic and transcriptional adaptations. Although the beneficial effects of exercise modalities on skeletal muscle structure and function in aging are well recognized, the exact cellular and molecular mechanisms underlying the influence of exercise have not been fully elucidated. We summarize the biochemical pathways involved in the progression and pathogenesis of sarcopenia and describe the beneficial effects of exercise training on the relevant signaling pathways associated with sarcopenia. This study briefly introduces current knowledge on the signaling pathways involved in the development of sarcopenia, effects of aerobic exercise on mitochondria-related parameters and mitochondrial function, and role of resistance exercise in the regulation of muscle protein synthesis against sarcopenia. This review suggested that the beneficial effects of exercise are still under-explored, and accelerated research will help develop better modalities for the prevention, management, and treatment of sarcopenia.


The role of imaging in catheter ablation of ventricular arrhythmias.
Late gadolinium enhancement cardiac magnetic resonance (LGE-CMR) and multidetector cardiac computed tomography (MDCT) have emerged as novel, fascinating imaging tools for arrhythmogenic substrate identification and characterization. The role of these techniques for aiding and guiding the catheter ablation of ventricular tachycardia, either as a complement or a surrogate of the electroanatomic map, has been rising in recent years. Integrating pixel signal intensity maps or wall thickness maps delivered from LGE-CMR or MDCT, respectively, into the navigation system has become a cornerstone for VT ablation procedures in a few centers of excellence around the world. The pre-procedure scar characterization offers some advantages, helping decide for the best procedure planning and approach; complete substrate identification and characterization, helping to focus electroanatomical mapping in regions of interest and also has a positive impact in procedure efficiency and outcomes. In the present article, we perform a review of the most practical aspects for using LGE-CMR or MDCT when a VT ablation procedure is planned, from the image acquisition to the integration into the navigation system, analyzing the current role of the LGE-CMR and MDCT for arrhythmogenic substrate characterization as well as for guiding VT ablation.


Prevention of pain on injection of propofol: a comparison of lidocaine with alfentanil.
We undertook a randomized, placebo-controlled, double-blind study to compare the use of alfentanil 1 mg and lidocaine 40 mg for the reduction of pain during injection of propofol. Eighty-nine patients were randomly allocated to one of three groups: Group L, lidocaine 40 mg added to 180 mg propofol; Group A, alfentanil 1 mg 30 s prior to propofol; or Group P, placebo (normal saline). The incidence of pain in the placebo group was 67%. Both treatment groups had a significantly lower incidence of pain than the placebo group (P < 0.002). There was no significant difference in the incidence of pain between the groups receiving lidocaine or alfentanil (13% and 24%, respectively). There was no significant difference in the induction dose of propofol between the groups. Fifty-two percent of patients who experienced pain at induction had recall of that pain in the recovery room. Alfentanil 1 mg and lidocaine 40 mg are both effective in reducing pain during injection of propofol.


Imaging Techniques for Clinical Burn Assessment with a Focus on Multispectral Imaging.
Significance: Burn assessments, including extent and severity, are some of the most critical diagnoses in burn care, and many recently developed imaging techniques may have the potential to improve the accuracy of these evaluations. Recent Advances: Optical devices, telemedicine, and high-frequency ultrasound are among the highlights in recent burn imaging advancements. We present another promising technology, multispectral imaging (MSI), which also has the potential to impact current medical practice in burn care, among a variety of other specialties. Critical Issues: At this time, it is still a matter of debate as to why there is no consensus on the use of technology to assist burn assessments in the United States. Fortunately, the availability of techniques does not appear to be a limitation. However, the selection of appropriate imaging technology to augment the provision of burn care can be difficult for clinicians to navigate. There are many technologies available, but a comprehensive review summarizing the tissue characteristics measured by each technology in light of aiding clinicians in selecting the proper device is missing. This would be especially valuable for the nonburn specialists who encounter burn injuries. Future Directions: The questions of when burn assessment devices are useful to the burn team, how the various imaging devices work, and where the various burn imaging technologies fit into the spectrum of burn care will continue to be addressed. Technologies that can image a large surface area quickly, such as thermography or laser speckle imaging, may be suitable for initial burn assessment and triage. In the setting of presurgical planning, ultrasound or optical microscopy techniques, including optical coherence tomography, may prove useful. MSI, which actually has origins in burn care, may ultimately meet a high number of requirements for burn assessment in routine clinical use.


Management strategies for cardiogenic shock.
Cardiogenic shock remains the most serious complication of acute MI, with an incidence of 6 to 8% and a 30-day mortality rate that remains close to 50%. While cardiogenic shock is due primarily to left ventricular failure, other causes such as acute mitral regurgitation and ventricular septal rupture must always be considered as emergency surgery may be life saving. The purpose of this review is to summarize recent advances in the care of these critically ill patients including the consideration of etiology and pathophysiology as well as the influence of age and adjunctive therapies. Early revascularization is now an American College of Cardiology/American Heart Association guideline class 1 indication for percutaneous coronary intervention (PCI) particularly for younger patients in cardiogenic shock. Recent studies suggest there may also be a benefit in elderly patients with cardiogenic shock. Prompt triage of all patients in cardiogenic shock for early angiography, intra-aortic balloon pump counterpulsation, and early revascularization with PCI or bypass surgery is now the preferred management strategy.


Angiotensin II stimulates system y+ and cationic amino acid transporter gene expression in cultured vascular smooth muscle cells.
The effect of angiotensin II (Ang II) on the transport of cationic amino acids has been examined in vascular smooth muscle cells (VSMC) isolated from rat aortae. Ang II stimulated the uptake rates of radiolabeled arginine and lysine in a time- and concentration-dependent manner. The stimulated arginine uptake could be blocked by pretreatments with cycloheximide and actinomycin D or co-treatment with valsartan, an antagonist specific for Ang II receptor subtype-1. The modulation by Ang II was bidirectional as the efflux of arginine was also stimulated, 5-fold over basal. Using reverse transcription-coupled polymerase chain reaction methodology, a partial cDNA with 94% sequence identity to that of cationic amino acid transporter subtype-1 (CAT-1) of mouse fibroblasts was obtained from VSMC. This sequence also exhibited 14 base changes compared with the sequence of ecotropic retrovirus receptor (ERR)/CAT-1 from rat hepatoma. Northern analyses with this partial CAT-1 cDNA and CAT-2 cDNA of mouse T-lymphocytes showed that Ang II rapidly stimulated the expression of both CAT-1 and CAT-2 in VSMC. Both signals peaked at 2 h after exposure to Ang II. The CAT-1 signal decayed over the next 6 h to levels 3-fold above basal, which are maintained up until 24 h. The induced CAT-2 mRNA concentration also decayed rapidly but increased again between 16 and 24 h to levels comparable with those observed at 2 h.


Search for Antimicrobial Activity Among Fifty-Two Natural and Synthetic Compounds Identifies Anthraquinone and Polyacetylene Classes That Inhibit Mycobacterium tuberculosis.
Drug-resistant tuberculosis threatens to undermine global control programs by limiting treatment options. New antimicrobial drugs are required, derived from new chemical classes. Natural products offer extensive chemical diversity and inspiration for synthetic chemistry. Here, we isolate, synthesize and test a library of 52 natural and synthetic compounds for activity against Mycobacterium tuberculosis. We identify seven compounds as antimycobacterial, including the natural products isobavachalcone and isoneorautenol, and a synthetic chromene. The plant-derived secondary metabolite damnacanthal was the most active compound with the lowest minimum inhibitory concentration of 13.07 μg/mL and a favorable selectivity index value. Three synthetic polyacetylene compounds demonstrated antimycobacterial activity, with the lowest MIC of 17.88 μg/mL. These results suggest new avenues for drug discovery, expanding antimicrobial compound chemistries to novel anthraquinone and polyacetylene scaffolds in the search for new drugs to treat drug-resistant bacterial diseases.


Analysis of cDNAs of the proto-oncogene c-src: heterogeneity in 5' exons and possible mechanism for the genesis of the 3' end of v-src.
To further characterize the gene structure of the proto-oncogene c-src and the mechanism for the genesis of the v-src sequence in Rous sarcoma virus, we have analyzed genomic and cDNA copies of the chicken c-src gene. From a cDNA library of chicken embryo fibroblasts, we isolated and sequenced several overlapping cDNA clones covering the full length of the 4-kb c-src mRNA. The cDNA sequence contains a 1.84-kb sequence downstream from the 1.6-kb pp60c-src coding region. An open reading frame of 217 amino acids, called sdr (src downstream region), was found 105 nucleotides from the termination codon for pp60c-src. Within the 3' noncoding region, a 39-bp sequence corresponding to the 3' end of the RSV v-src was detected 660 bases downstream of the pp60c-src termination codon. The presence of this sequence in the c-src mRNA exon supports a model involving an RNA intermediate during transduction of the c-src sequence. The 5' region of the c-src cDNA was determined by analyzing several cDNA clones generated by conventional cloning methods and by polymerase chain reaction. Sequences of these chicken embryo fibroblast clones plus two c-src cDNA clones isolated from a brain cDNA library show that there is considerable heterogeneity in sequences upstream from the c-src coding sequence. Within this region, which contains at least 300 nucleotides upstream of the translational initiation site in exon 2, there exist at least two exons in each cDNA which fall into five cDNA classes. Four unique 5' exon sequences, designated exons UE1, UE2, UEX, and UEY, were observed. All of them are spliced to the previously characterized c-src exons 1 and 2 with the exception of type 2 cDNA. In type 2, the exon 1 is spliced to a novel downstream exon, designated exon 1a, which maps in the region of the c-src DNA defined previously as intron 1. Exon UE1 is rich in G+C content and is mapped at 7.8 kb upstream from exon 1. This exon is also present in the two cDNA clones from the brain cDNA library. Exon UE2 is located at 8.5 kb upstream from exon 1. The precise locations of exons UEX and UEY have not been determined, but both are more than 12 kb upstream from exon 1. The existence and exon arrangements of these 5' cDNAs were further confirmed by RNase protection assays and polymerase chain reactions using specific primers. Our findings indicate that the heterogeneity in the 5' sequences of the c-src mRNAs results from differential splicing and perhaps use of distinct initiation sites. All of these RNAs have the potential of coding for pp60c-src, since their 5' exons are all eventually joined to exon 2.


Migrant children--a challenge for European paediatricians.
Each year, tens of thousands of children seek asylum in Europe. Irregular immigration, family reunification programmes and labour migrants contribute additional large populations to the migrant scene in Europe. Paediatricians struggle to find solutions to their needs of health and medical care within different societal and political contexts on a common platform of children's rights and medical ethics. Migrant children have important health and protection needs. Paediatricians can improve the health and well-being of migrant children by assuring access to paediatric healthcare regardless of the child's legal status. Paediatricians can play an important role in advocating for migrant children in their local community, through concerted activities of national paediatric societies, and through the international paediatric community. National paediatric societies need to put migrant children high on their agenda in discussions with policy and decision makers. Initiatives are needed to create platforms within the international paediatric community where practical strategies to the multitude of issues touched upon by migrant children can be exchanged and international advocacy organized.


Association between diet and periodontitis: a cross-sectional study of 10,000 NHANES participants.
Periodontitis is a major cause of tooth loss globally. Risk factors include age, smoking, and diabetes. Intake of specific nutrients has been associated with periodontitis risk but there has been little research into the influence of overall diet, potentially more relevant when formulating dietary recommendations. We aimed to investigate potential associations between diet and periodontitis using novel statistical techniques for dietary pattern analysis. Two 24-h dietary recalls and periodontal examination data from the cross-sectional US NHANES, 2009-2014 (n = 10,010), were used. Dietary patterns were extracted using treelet transformation, a data-driven hierarchical clustering and dimension reduction technique. Associations between each pattern [treelet component (TC)] and extent of periodontitis [proportion of sites with clinical attachment loss (CAL) ≥ 3 mm] were estimated using robust logistic quantile regression, adjusting for age, sex, ethnicity, education level, smoking, BMI, and diabetes. Eight TCs explained 21% of the variation in diet, 1 of which (TC1) was associated with CAL extent. High TC1 scores represented a diet rich in salad, fruit, vegetables, poultry and seafood, and plain water or tea to drink. There was a substantial negative gradient in CAL extent from the lowest to the highest decile of TC1 (median proportion of sites with CAL ≥ 3 mm: decile 1 = 19.1%, decile 10 = 8.1%; OR, decile 10 compared with decile 1: 0.67; 95% CI: 0.46, 0.99). Most dietary patterns identified were not associated with periodontitis extent. One pattern, however, rich in salad, fruit, and vegetables and with plain water or tea to drink, was associated with lower CAL extent. Treelet transformation may be a useful approach for calculating dietary patterns in nutrition research.


9-(5',6'-dideoxy-beta-D-ribo-hex-5'-ynofuranosyl)adenine, a novel irreversible inhibitor of S-adenosylhomocysteine hydrolase.
The acetylenic analogue of adenosine 9-(5',6'-dideoxy-beta-D-ribo-hex-5'-ynofuranosyl)adenine has been synthesized, and its behavior as an inhibitor of bovine S-adenosylhomocysteine hydrolase has been examined. Incubation of the enzyme with excess inhibitor caused a time-dependent, irreversible inactivation of the enzyme that was accompanied by the reduction of two equivalents of NAD+ to NADH and the loss of the two remaining equivalents of NAD+. With use of radiolabeled inhibitor, it was established that 4 equiv of the acetylenic analog bind irreversibly to the enzyme and that 4 equiv were required to inactivate the enzyme completely. The inactivated enzyme could not be reactivated by incubation with NAD+. Denaturation studies revealed that 2 equiv of the inhibitor are bound more tightly to the enzyme than the remainder, suggesting the formation of a covalent linkage between the oxidized inhibitor and the enzyme. The putative covalent linkage was found to be acid sensitive but stable to mild base. The linkage could not be stabilized by treatment of the enzyme-inhibitor complex with either borohydride or cyanoborohydride. A Kl of 173 nM was measured for the inhibitor, making it one of the more potent inhibitors that have been reported. The enzyme used in these studies was isolated by modification of an affinity chromatography method reported by Narayanan and Borchardt [(1988) Biochim. Biophys. Acta 965, 22-28]. The affinity chromatography unexpectedly led to the isolation of two forms of the enzyme. The major form contained 4.0 mol of nucleotide cofactor/mol of enzyme tetramer, while the minor form carried only 2.0 mol/tetramer.


Residual Deformity and Outcome of Ambulatory Adults With Cerebral Palsy: A Long-term Longitudinal Assessment.
Advances in pediatric orthopaedic care have improved mobility and function for children with cerebral palsy (CP) as mobility declines from adolescence into adulthood. The long-term effectiveness of modern orthopaedic care is not widely reported. This study aimed to report the pediatric orthopaedic surgical burden, residual deformities, and outcomes using objective evidence of mobility in ambulatory adults with CP. An institutional review board-approved prospective cohort study was performed in ambulatory adults with CP between 25 and 45 years, who had an adolescent gait analysis. Orthopaedic interventions were reviewed, and adolescent and adult gait analyses were compared using paired 2-tailed t tests. Adults were categorized by the presence of no, mild, or severe residual deformities in rotation, crouch, stiff knee, equinus, and foot deformity. Of 106 adults with CP, Gross Motor Function Classification System (GMFCS) distribution was grade I (22%), II (50%), III (23%), and IV (5%). Sixty-one males and 45 females were tested. The average age was 30±4 years with follow-up of 13±4 years since previous analysis; 279 surgical events (1165 procedures) were performed with a mean per patient of 2.6 events and 11 procedures. Comm on procedures were gastrocsoleus complex (88%) and hamstring lengthening (79%). The mean gait deviation index at adolescent and adult visit were 72.7±13 and 72.3±13 (P=0.78). Mean gait velocity at the adolescent visit was 85±27 and 79±31 cm/s at adult visit (P=0.02). Both gait deviation index and gait velocity change were clinically insignificant. Fifty-seven adults (81 limbs, 54%) had mild residual deformities. Residual hip internal rotation, pes planovalgus, and crouch gait were common. Severe deformities impacting function or causing pain were present in 11 participants (14 limbs, 10%). Seven of the 11 adults with severe deformities were worse compared with their adolescent evaluation; 4 were unchanged. Correcting deformities before adulthood has lasting stability with little functional loss in most ambulatory young adults with CP. Increasing deformity after adolescence can occur in young adults but is uncommon. Level III.


Retinoic acid suppresses IL-17 production and pathogenic activity of γδ T cells in CNS autoimmunity.
Retinoic acid (RA) in the steady state enhances induction of Foxp3(+) regulatory T (Treg) cells and inhibits differentiation of Th1 and Th17 cells, thereby maintaining tolerance, but can in inflammatory conditions promote effector Th1 and Th17 cells that mediate inflammation. IL-17-producing γδ T cells have recently been shown to have a major pathogenic role in autoimmune diseases. Here, we examined the immunomodulatory effects of RA on γδ T cells. We found that RA had a dramatic suppressive effect on IL-17A and IL-17F production by γδ T cells stimulated with IL-1β and IL-23. RA suppressed RORγt, IL-1R and IL-23R expression in γδ T cells. Treatment of mice with RA suppressed IL-17 production by γδ T cells in vivo. Furthermore, treatment of T cells with RA attenuated their ability to induce disease in experimental autoimmune encephalomyelitis (EAE), a murine model for multiple sclerosis. This was associated with a reduction in the number of central nervous system-infiltrating γδ T cells, but also CD4(+) T cells that produced IL-17A, IL-17F or GM-CSF. Interestingly, treatment of γδ T cells with RA or removal of γδ T cells from a bulk population of T cells significantly reduced their capacity to induce EAE, demonstrating a critical role for γδ T cells in promoting pathogenic Th17 cells. Our findings demonstrate that the anti-inflammatory properties of RA are mediated in part by suppressing STAT3-mediated activation of cytokine production and cytokine receptor expression in γδ T cells, which suppresses their ability to activate Th17 cells.


Mitochondria-Targeted Reaction-Based Fluorescent Probe for Hydrogen Sulfide.
In this study, we developed a turn-on mitochondria-targeting hydrogen sulfide, "probe 1", based on the selective thiolysis of 7-nitro-1,2,3-benzoxadiazole amine moiety attached to the piperazine-based naphthalimide scaffold. Probe 1 exhibited excellent properties with 68-fold fluorescence enhancement, a low detection limit (2.46 μM), a low cytotoxicity, and a good selectivity toward hydrogen sulfide. The success of intracellular imaging indicated that probe 1 could be used in further applications for the investigation of biological functions and pathological roles of H2S in living systems.


Use of MindMapper software for research domain mapping.
The application of concept mapping software to facilitate the first steps of the research review process is discussed in relation to other software programs currently used for research synthesis. MindMapper software was used to develop a strategy for organizing the results of a comprehensive literature search into discrete categories with relationships among concepts graphically displayed to reveal the structure of the research domain. A Mind Map was developed for the scholarly literature on Web-based consumer health information. A second Mind Map examined the subconcept of barriers to consumer use of the Internet for health information. MindMapper has many features that facilitate description of the breadth and depth of literature in a domain of inquiry. It also facilitates identification of the number and nature of studies underpinning mapped relationships among concepts, thus laying the groundwork for systematic research reviews and meta-analyses.


The effect of xanthine derivatives on red blood cells: microelectrophoretic studies.
The effect of xanthine derivatives on the variation of surface charges of red blood cells has been investigated. Results of mobility curves showed that the derivatives increase the surface charges on the cells, whereas there is little effect on the surface charges of liposomes.


Transfer factors of some selected radionuclides (radioactive Cs, Sr, Mn, Co and Zn) from soil to leaf vegetables.
Transfer factors of radionuclides from soil to leaf vegetables (cabbage, Chinese cabbage, komatsuna, spinach and lettuce) have been studied by radiotracer experiments using Andosol as a representative of Japanese soils. The transfer factors of radioactive Cs, Sr, Mn, Co and Zn for edible parts of vegetables (average of five vegetables) were 0.11, 0.24, 0.61, 0.05 and 0.52, respectively. These values should be used in safety assessment for Japanese agricultural environment. The transfer factors of Mn, Co and Zn for spinach were higher than those for the other vegetables. The transfer factors of Cs for different organs of the leaf vegetables were rather homogeneous. The transfer factors of Sr and Mn were higher for older (outer) leaves than younger (inner) ones. In contrast to Sr and Mn, transfer factors of Zn for younger leaves were higher than those for older ones. The distribution ratios of the elements between soil-solution and soil were in the order Sr > Mn > Cs > Co > Zn, whereas the distribution ratios of the elements between plant and soil-solution were in the order Zn > Cs > Mn > Co > Sr. These results indicate that the selectivity for Sr by plants from the soil-solution was low and that for Zn was very high.