Heparin induces rat aorta relaxation via integrin-dependent activation of muscarinic M3 receptors.
Previous reports have shown that heparin may promote human hypotension and vascular relaxation by elevation of NO levels through unclear mechanisms. We hypothesized that endothelial muscarinic M(3) receptor activation mediates the heparin-induced vasodilation of rat aortic rings. The experiments were carried out using unfractionated heparin extracted from bovine intestinal mucosa, which elicited an endothelium and NO-dependent relaxation of aortic segments with maximal potency and efficacy (EC(50): 100±10 μmol/L; E(max): 41±3%). Atropine and 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide inhibitors reduced the heparin-dependent relaxation, indicating that M(3) muscarinic receptor is involved in this phenomenon. However, no direct binding of heparin to muscarinic receptors was observed. More importantly, studies performed using the arginine-glycine-aspartic acid peptide and 1-(1,1-dimethylethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-day]pyrimidin-4-amine, an Src family inhibitor, reduced by 51% and 73% the heparin-dependent relaxation, respectively, suggesting the coupling of heparin and M(3) receptor through extracellular matrix molecules and integrin. Furthermore, unfractionated heparin induced activation of focal adhesion protein kinase, Src, and paxillin. Finally, fluorescence resonance energy transfer approach confirmed the interaction of the M(3) receptor to integrin. Taken together, these data demonstrate the participation of M(3) receptor and integrin in heparin-dependent relaxation of vascular smooth muscle. These results provide new insights into the molecular mechanism and potential pharmacological action of heparin in vascular physiology.


Aspirin inhibits interleukin 1-induced prostaglandin H synthase expression in cultured endothelial cells.
Prostaglandin H (PGH) synthase (EC 184.108.40.206) is a key enzyme in the biosynthesis of prostaglandins, thromboxane, and prostacyclin. In cultured human umbilical vein endothelial cells, interleukin 1 (IL-1) is known to induce the synthesis of this enzyme, thereby raising the level of PGH synthase protein severalfold over the basal level. Pretreatment with aspirin at low concentrations (0.1-1 micrograms/ml) inhibited more than 60% of the enzyme mass and also the cyclooxygenase activity in IL-1-induced cells with only minimal effects on the basal level of the synthase enzyme in cells without IL-1. Sodium salicylate exhibited a similar inhibitory action whereas indomethacin had no apparent effect. Similarly low levels of aspirin inhibited the increased L-[35S]methionine incorporation into PGH synthase that was induced by IL-1 and also suppressed expression of the 2.7-kilobase PGH synthase mRNA. These results suggest that in cultured endothelial cells a potent inhibition of eicosanoid biosynthetic capacity can be effected by aspirin or salicylate at the level of PGH synthase gene expression. The aspirin effect may well be due to degradation of salicylate.


Novel Technique for Measuring Tissue Firmness within Tomato (Lycopersicon esculentum Mill.) Fruit.
Developmental changes of tomato (Lycopersicon esculentum) fruit tissues during maturation were analyzed by a physically defined method (stress-relaxation analysis). The tip of a conical probe connected to a load sensor was positioned on the cut surface of a sliced tomato fruit, and the decay of the imposed stress was monitored. Stress-relaxation data thus obtained were used for the calculation of three stress-relaxation parameters. Different zones within tomato fruit harvested at six different ripening stages were analyzed. One of the stress-relaxation parameters, minimum stress-relaxation time (T(0)), decreased as the fruits matured. The decrease in T(0) was first found in the core of the carpel junction within the endopericarp at the blossom end during the breaker stage. The decrease in T(0) progressed from the blossom end, through the equatorial region and finally throughout the shoulder, as the fruit matured. In mature green fruit, T(0) values within the placenta and the proximal carpel junction were lower than those by other parts of the fruit. For all measurements the maximum stress-relaxation time was not substantially changed during maturation, nor were their changes observed in different regions of the fruit. The observed relaxation rate was therefore correlated with softening. The results indicate that fruit softening may be physically associated with the stress-relaxation parameter, T(0), and the extent of softening is a function of position within the fruit. Decreases in T(0) value appear to be correlated with the reported regional variation in the appearance of polygalacturonase.


New Strategies in the Efficient Total Syntheses of Polycyclic Natural Products.
Polycyclic natural products are an inexhaustible source of medicinal agents, and their complex molecular architecture renders challenging synthetic targets where innovative and effective approaches for their rapid construction are urgently required. The total synthesis of polycyclic natural products has witnessed exponential progression along with the emergence of new synthetic strategies and concepts, such as sequential C-H functionalizations, radical-based transformations, and functional group pairing strategies. Our group exerts continued interest in the construction of bioactive and structurally complex natural products as well as evaluation of the mode of action of these molecules. In this Account, we will showcase how these new synthetic strategies are employed and guide our total synthesis endeavors.During the last two decades, a series of remarkable advances in C-H functionalization have led to the emergence of many new approaches to directly functionalize C-H bonds into useful functional groups. These selective transformations have provided a great opportunity for the step- and atom-economical construction of key fragments in complex molecule synthesis. We recently furnished the total syntheses for polycyclic natural products: incarviatone A, chrysomycin A, polycarcin V, and gilvocarcin V by employing a multiple C-H bond functionalization strategy. The polysubstituted benzene or naphthalene skeleton was constructed through sequential and site-selective C-H functionalizations from readily available simple starting materials, which reduced the number of steps and streamlined synthesis.Recently, we have also completed the total syntheses for a number of skeletally diverse tetracyclic Isodon diterpenoids inspired by their biogenesis and radical-based retrosynthetic disconnections. Radical transformations are strategically and tactically utilized in our syntheses, and radical-based reactions, including organo-SOMO catalysis, Birch reduction, regioselective 1,6-dienyne reductive cyclization, visible-light-mediated Schenck ene reaction, and photoradical-mediated late-stage skeletal rearrangement, play significant roles in our synthetic endeavors. Protecting-group-free and scalable syntheses are also built into our work to achieve the "ideal" synthesis. Furthermore, our synthetic work reveals that late-stage skeletal rearrangement through a photo radical process is possible in a biological setting in complement with nature's carbocation chemistry in complex natural product biosynthesis.Lycopodium alkaloids are a large family of structurally unique polycyclic natural products with impressive biological activities. Owing to their fascinating polycyclic architectures and diverse biological activities, these alkaloids have continued to serve as targets as well as inspirations for the synthetic community for decades. To access these bioactive natural products or natural product-like molecules for biological exploration and drug discovery, we applied a novel functional group pairing strategy to furnish the total syntheses for several Lycopodium alkaloids and obtained numerous skeletally diverse compounds with structural complexity comparable to natural products.


An overview on biologic medications and their possible role in apical periodontitis.
Apical periodontitis (AP) is the expression of a deficient balance between infection and the host immune response. If reducing the bacterial load from the root canal and preventing its reinfection may lead to clinical success, then the integrity of the nonspecific immune system has a relevant influence on the outcome of endodontic treatment. Compromised immune systems and/or genetic alterations of the host's response may as well play an important role on the development, progression, and healing of AP. Thus, immunomodulatory drugs might have the potential to influence both the severity of AP and the outcome of endodontic treatment. Biologic medications are a new class of drugs of monoclonal antibodies or fusion proteins that include fragments of a peculiar cytokine receptor. Specific inflammatory molecules or cells, such as tumor necrosis factor, interleukins, and T or B cells, are the selective targets of these drugs. They modulate the altered immune response and perform an important role in the short-term treatment of chronic inflammatory diseases such as rheumatoid arthritis, refractory Crohn disease, or ulcerative colitis. Despite the clinical positive outcomes and their widespread use, the consequences of administering biologic medications on the development of the dental diseases have not been adequately investigated. The aim of this review was to give an overview of biologic medications, their composition, their mechanisms of action, and their possible implications on endodontic and other dental diseases.


Manganese(II)-containing MRI contrast agent employing a neutral and non-macrocyclic ligand.
The ligand N,N'-bis(2-pyridylmethyl)-bis(ethylacetate)-1,2-ethanediamine (debpn) coordinates divalent transition metal ions in either a pentadentate or hexadentate fashion. The coordination number correlates with the ionic radius of the metal ion, with larger cations being heptacoordinate as assessed by solid-state analysis. With Mn(II), the debpn ligand is hexadentate and remains bound to the oxophilic metal ion, even when dissolved in water. The ligand's incomplete coordination of the manganous ion allows water molecules to coordinate to the metal center. These two properties, coupled with the high paramagnetism associated with the S = 5/2 metal center, enable [Mn(debpn)(H(2)O)](ClO(4))(2) to serve as a stable and effective magnetic resonance imaging contrast agent despite the ligand's lack of both a macrocyclic component and an anionic charge.


The expedited discharge of patients with multiple traumatic rib fractures is cost-effective.
Rib fractures are a cause of significant morbidity and mortality in trauma patients. It is well documented that optimizing pain control, mobilization, and respiratory care decreases complications. However, the impact of these interventions on hospital costs and length of stay is not well defined. We hypothesized patients with multiple rib fractures can be discharged within three hospital days resulting in decreased hospital costs. A retrospective review of adult patients (≥18yrs) admitted to our Level 1 trauma center (2011-2013) with ≥2 rib fractures was performed. Patients were excluded if they were intubated, admitted to the ICU, required chest tube placement, or sustained significant multi-system trauma. (n = 202) Demographics, clinical characteristics, hospital costs, and outcome data were analyzed. Patients discharged within three hospital days of admission were considered to have achieved expedited discharge (ED). Univariate and multivariate analyses determined predictors of failure to achieve ED. A p value of <0.05 was considered significant. Study patients (n = 202) were 60 (SD = 19) years of age with an injury severity score (ISS) of 10 (SD = 5), and 4 (SD = 2) rib fractures. Of 202 patients, 127 (63%) achieved ED while 75 (37%) did not. No differences in chest AIS, ISS, smoking status or history of pulmonary disease were identified between the two groups (all p > 0.05). Average LOS (2 (SD = 1) vs. 7 (SD = 4) days; p < 0.001) and hospital costs ($2865 (SD = 1200) vs. $6085 (SD = 3033)); p < 0.001). were lower in the ED group A lower percentage of ED patients required placement in rehabilitation facilities (6% vs. 48%; p < 0.001). There were no readmissions within 30 days in either group. After controlling for potential confounding variables, multiple variable logistic regression analysis revealed that advancing age (OR 1.05 per year, 1.02-1.07) independently predicted failure to achieve ED. The majority of patients admitted to the hospital with multiple rib fractures can be discharged within three days. This expedited discharge results in significant cost savings to the hospital. Early identification of patients who cannot meet the goal of expedited discharge can facilitate improvement in management strategies.


[Characterization of the HUMTPOX and HUMCSF1PO polymorphic systems in a sample of the Parma population].
A population study in a sample from the Province of Parma was carried out to investigate the polymorphic systems HUMTPOX and HUMCSF1PO. Separation of duplex PCR co-amplified fragments was performed by denaturing PAGE followed by silver staining; seven and six allelic variants respectively were found and frequency distribution was in good agreement with previous studies on Caucasian. The power of Discrimination (PD) was 0.82 and 0.88 respectively and the chance of exclusion (MEC) 0.401 and 0.502; the robustness of the polymorphic systems and informative strength, though not particularly high, suggest the use of these polymorphisms in forensic investigations.


[Sexual and reproductive function in female liver transplant recipients].
Abnormalities in sexual and reproductive functions are common in women with end-stage liver disease and may be reversible after liver transplantation. Evaluate sexual and reproductive function in female recipients of liver transplantation at the Federal University of Parana, Curitiba, PR, Brazil. Between September 1991 and December 2001 94 women underwent liver transplantation at "Hospital de Clinicas" of the Federal University of Parana. Twenty-eight female recipients (mean age 44.17 +/- 13.60 years old) fulfilled the following inclusion criteria: age > or = 16 years at liver transplant, post-transplant survival > or = 6 months, be alive and in regular follow-up at our Institution during the period of the study, and agreed to participate of the study. Medical records were reviewed and all subjects were answered by a questionnaire covering the following issues: timing and pattern of menstruation before and after transplant, sexual activity and contraceptive practices before and after transplant, pregnancy after transplant, frequency of cervical cytology and occurrence of gynecological malignant neoplasia after transplant, and aspects of sexuality domain after liver transplantation. The median post-transplant follow-up was 36.5 months (range, 6 to 110 months) and the main indication for liver transplantation was hepatitis C (25%). All patients had normal liver function tests. Excluding six patients who underwent hysterectomy or were in postmenopausal period, 13 of 22 women (59.1%) reported amenorrhea in the year before transplantation. After liver transplantation, 19 of 22 patients (86.4%) promptly recovered menstrual function, after a median period of 1 month (range, 1 to 7 months). Normalization of menstrual function occurred in all women with age 45 or younger. About 71.4% of 28 women were sexually active after transplantation and 70% indicated satisfaction with their relationship. Four successful pregnancies (one gemelar) occurred in three patients and five healthy live-born infants have been delivered, all full term. Most of the patients had cervical cytology at least annually in the post-transplant period. One case of endometrial carcinoma was diagnosed in a 64 year old woman, three years after transplantation and was successfully treated by panhysterectomy. Most of female liver transplant recipients of child-bearing age recover menstrual function a few months after transplantation and successful pregnancy may occur. As libido and fertility return promptly after liver transplantation, patients should be counseled on safe contraception practices. Most of liver transplant recipients are sexually active and feel satisfied about their relationship. There is a good compliance of patients regarding screening for cervical neoplasm.


[Two cases of pulmonary lymphangioleiomyomatosis].
We report two cases of pulmonary lymphangioleiomyomatosis with recurrent spontaneous pneumothorax. The cases were diagnosed by open lung biopsy. Case 1 was a 38-year-old woman, whose chest X-ray showed linear and reticular shadows. Lung tissue was negative for hormone receptors. She was treated with tamoxifen, but developed allergic symptoms. Her condition gradually deteriorated despite oxygen and progesterone therapy. Case 2 was a 41-year-old woman, whose chest X-ray showed a reticular shadows and slight overinflation. Hormone receptors were not examined, but the disease did not progress with oxygen, progesterone and GnRH agonist (Sprecur) therapy.


The Distinct Roles of Class I and II RPD3-Like Histone Deacetylases in Salinity Stress Response.
Histone acetylation is an essential process in the epigenetic regulation of diverse biological processes, including environmental stress responses in plants. Previously, our research group identified a histone deacetylase (HDAC) inhibitor (HDI) that confers salt tolerance in Arabidopsis (Arabidopsis thaliana). In this study, we demonstrate that class I HDAC (HDA19) and class II HDACs (HDA5/14/15/18) control responses to salt stress through different pathways. The screening of 12 different selective HDIs indicated that seven newly reported HDIs enhance salt tolerance. Genetic analysis, based on a pharmacological study, identified which HDACs function in salinity stress tolerance. In the wild-type Columbia-0 background, hda19 plants exhibit tolerance to high-salinity stress, while hda5/14/15/18 plants exhibit hypersensitivity to salt stress. Transcriptome analysis revealed that the effect of HDA19 deficiency on the response to salinity stress is distinct from that of HDA5/14/15/18 deficiencies. In hda19 plants, the expression levels of stress tolerance-related genes, late embryogenesis abundant proteins that prevent protein aggregation and positive regulators such as ABI5 and NAC019 in abscisic acid signaling, were induced strongly relative to the wild type. Neither of these elements was up-regulated in the hda5/14/15/18 plants. The mutagenesis of HDA19 by genome editing in the hda5/14/15/18 plants enhanced salt tolerance, suggesting that suppression of HDA19 masks the phenotype caused by the suppression of class II HDACs in the salinity stress response. Collectively, our results demonstrate that HDIs that inhibit class I HDACs allow the rescue of plants from salinity stress regardless of their selectivity, and they provide insight into the hierarchal regulation of environmental stress responses through HDAC isoforms.


Catalytic addition of terminal alkynes to carbodiimides by half-sandwich rare earth metal complexes.
The catalytic addition of terminal alkynes to carbodiimides has been achieved for the first time by use of half-sandwich rare earth metal complexes, such as {Me2Si(C5Me4)(NPh)}Y(CH2SiMe3)(THF)2, which offers a straightforward, atom-economic route to the N,N'-disubstituted propiolamidines which contain a conjugated C-C triple bond, a new family of amidines which were difficult to prepare by other means. A rare earth metal amidinate species was confirmed to be a true catalytic species in this process, thus demonstrating for the first time that an amidinate unit, though being often used as an ancillary ligand for various organometallic complexes, can itself participate in a catalytic reaction under appropriate conditions.


Characterization and inhibition of norovirus proteases of genogroups I and II using a fluorescence resonance energy transfer assay.
Noroviruses are the major cause of food- or water-borne gastroenteritis outbreaks in humans. The norovirus protease that cleaves a large viral polyprotein to nonstructural proteins is essential for virus replication and an attractive target for antiviral drug development. Noroviruses show high genetic diversity with at least five genogroups, GI-GV, of which GI and GII are responsible for the majority of norovirus infections in humans. We cloned and expressed proteases of Norwalk virus (GI) and MD145 virus (GII) and characterized the enzymatic activities with fluorescence resonance energy transfer substrates. We demonstrated that the GI and GII proteases cleaved the substrates derived from the naturally occurring cleavage site in the open reading frame (ORF) 1 of G1 norovirus with similar efficiency, and that enzymatic activity of both proteases was inhibited by commercial protease inhibitors including chymostatin. The interaction of chymostatin to Norwalk virus protease was validated by nuclear magnetic resonance (NMR) spectroscopy.


Resolving the feeding difficulties associated with non-organic failure to thrive.
Since 1991 The Children's Society's Infant Support Project has been working in Wiltshire with the families of under 3 year olds identified as failing to thrive non-organically. Research has shown that the only common factor in children with this problem is the presence of feeding difficulties. The service concentrates on helping parents/caregivers to resolve the problems which are allowing the difficulties to continue. The multidisciplinary staff team work on a domiciliary basis, using working practices founded on social learning theory. A recent evaluation indicated that for more than two-thirds of the children with whom the project had worked, a satisfactory or very satisfactory outcome was achieved. Funding for the service is received from the Health Commission for Wiltshire and Bath, Wiltshire Social Services Department and the Children's Society.


PET Imaging for Dynamically Monitoring Neuroinflammation in APP/PS1 Mouse Model Using [18F]DPA714.
Background: In the pathogenesis of Alzheimer's disease (AD), microglia play an increasingly important role. Molecular imaging of neuroinflammatory targeting microglia activation and the high expression of 18-kDa translocator protein (TSPO) has become a hot topic of research in recent years. Dynamic monitoring neuroinflammation is crucial for discovering the best time point of anti-inflammatory therapy. Motivated by this, Positron emission tomography (PET) imaging in an APP/PS1 mouse model of AD, using 18F-labeled DPA-714 to monitor microglia activation and neuroinflammation, were performed in this paper. Methods: We prepared [18F]DPA714 and tested the biological characteristics of the molecular probe in normal mice. To obtain a higher radiochemical yield, we improved the [18F]-fluorination conditions in the precursor dosage, reaction temperature, and synthesis time. We performed [18F]DPA714 PET scanning on APP/PS1 mice at 6-7, 9-10, 12-13, and 15-16 months of age, respectively. The same experiments were conducted in Wild-type (Wt) mice as a control. Referring to the [18F]DPA714 concentrated situation in the brain, we performed blocking experiments with PK11195 (1 mg/kg) in 12-13-months-old APP/PS1 mice to confirm the specificity of [18F]DPA714 for TSPO in the APP/PS1 mice. Reconstructed brain PET images, fused with the Magnetic Resonance Imaging (MRI) template atlas, and the volumes of interests (VOIs) of the hippocampus and cortex were determined. The distribution of [18F]DPA714 in the brain tissues of 15-16-months-old APP/PS1 and Wt mice were studied by immunofluorescence staining. Results: Through the reaction of 18F, with 2 mg precursor dissolved in 300 ul acetonitrile at 105°C for 10 min, we obtained the optimal radiochemical yield of 42.3 ± 5.1% (non-decay correction). Quantitative analysis of brain PET images showed that the [18F]DPA714 uptake in the cortex and hippocampus of 12-13-months-old APP/PS1 mice was higher than that of the control mice of the same age (cortex/muscle: 2.77 ± 0.13 vs. 1.93 ± 0.32, p = 0.0014; hippocampus/muscle: 3.33 ± 0.10 vs. 2.10 ± 0.35, p = 0.0008). The same significant difference was found between 15- and 16-months-old APP/PS1 mice (cortex/muscle: 2.64 ± 0.14 vs. 1.86 ± 0.52, p=0.0159; hippocampus/muscle: 2.89 ± 0.53 vs. 1.77 ± 0.48, p = 0.0050). Immunofluorescence staining showed that the activation of microglia and the level of TSPO expression in the cortex and hippocampus of APP/PS1 mice were significantly higher than Wt mice. Conclusion: [18F]DPA714, a molecular probe for targeting TSPO, showed great potential in monitoring microglia activation and neuroinflammation, which can be helpful in discovering the best time point for anti-inflammatory therapy in AD.


Proportional integral feedback control of ultrafiltration rate in hemodialysis.
Most hemodialysis patients without residual kidney function accumulate fluid between dialysis session that needs to be removed by ultrafiltration. Ultrafiltration usually results in a decline in relative blood volume (RBV). Recent epidemiological research has identified RBV ranges that were associated with significantly better survival. The objective of this work was to develop an ultrafiltration controller to steer a patient's RBV trajectory into these favorable RBV ranges. We designed a proportional-integral feedback ultrafiltration controller that utilizes signals from a device that reports RBV. The control goal is to attain the RBV trajectory associated with improved patient survival. Additional constraints such as upper and lower bounds of ultrafiltration volume and rate were realized. The controller was evaluated in in silico and ex vivo bench experiments, and in a clinical proof-of-concept study in two maintenance dialysis patients. In all tests, the ultrafiltration controller performed as expected. In the in silico and ex vivo bench experiments, the controller showed robust reaction toward deliberate disruptive interventions (e.g. signal noise; extreme plasma refill rates). No adverse events were observed in the clinical study. The ultrafiltration controller can steer RBV trajectories toward desired RBV ranges while obeying to a set of constraints. Prospective studies in hemodialysis patients with diverse clinical characteristics are warranted to further explore the controllers impact on intradialytic hemodynamic stability, quality of life, and long-term outcomes.


Glutathione and the antioxidant potential of binary mixtures with flavonoids: synergisms and antagonisms.
Polyphenols are able to trap free radicals, which contributes to their known antioxidant capacity. In plant extracts, these secondary metabolites may act in concert, in a way that their combined activities will be superior to their individual effects (synergistic interaction). Several polyphenols have demonstrated clear antioxidant properties in vitro, and many of their biological actions have been attributed to their intrinsic reducing capabilities. As so, the intake of these compounds at certain concentrations in the diet and/or supplementation may potentiate the activity of reduced form glutathione (GSH), thus better fighting oxidative stress. The aim of this work was to predict a structure-antioxidant activity relationship using different classes of flavonoids and to assess, for the first time, possible synergisms and antagonisms with GSH. For these purposes a screening microassay involving the scavenging of DPPH• was applied. In general, among the tested compounds, those lacking the catechol group in B ring showed antagonistic behaviour with GSH. Myricetin displayed additive effect, while quercetin, fisetin, luteolin, luteolin-7-O-glucoside, taxifolin and (+)-catechin demonstrated synergistic actions. Furthermore, adducts formed at C2' and C5' of the B ring seem to be more important for the antioxidant capacity than adducts formed at C6 and C8 of the A ring.


Pyrroles as effective agents for blocking amine nitrosation.
The ability of 10 pyrroles to block the acidic nitrosation of morpholine has been determined by using an assay that measures their effectiveness in the presence of a 10-fold excess of amine. The log (% N-nitrosomorpholine) formed is a linear function of blocking agent concentrations ranging from 0.0 to 1.5 times the equivalents of initial nitrite. The negative slopes of these plots allow a ranking of the effectiveness of the blocking agent. Several of the pyrroles have been found to be much more effective than established blocking agents such as ascorbic acid. The following order of blocking ability has been determined: 2,5-dimethylpyrrole = 1-benzyl-2,5-dimethylpyrrole much greater than 4-methylcatechol greater than ascorbic acid = 1,2-phenylenediamine = pyrrole greater than 1,2,5-tribenzylpyrrole = 1-benzylpyrrole = octamethylporphine much greater than ammonium sulfamate = hydrazine = 2,5-diphenylpyrrole much much greater than beta-nicotyrine greater than 2-pyrrolecarboxylic acid. Pyrroles give complex mixtures devoid of N-nitroso compounds upon nitrosation.


Platelet-to-Lymphocyte Ratio as a Potential Indicator of Infection-Associated Emergency Visits of Renal Transplant Recipients.
Infection-associated emergency department use in renal transplant recipients has been increasing as solid-organ transplant has become a more common treatment method for chronic kidney failure. Platelet-to-lymphocyte ratio has been demonstrated to be significantly elevated in nosocomial infections in patients treated at intensive care units and is positively correlated with duration of hospital stay. In this study, we aimed to determine whether the platelet-to-lymphocyte ratio could be used as an indicator of infection in renal transplant patients presenting to emergency departments. This case-control retrospective study included data from between May 2015 and February 2018. We used the patient information management system to review patient medical records and laboratory test results of study participants. Our study included 156 adults in the patient group (recipients with infection) and 170 adults in the control group (recipients without infection). We observed significant differences between patient and control groups in terms of the number of days of hospital stay; leukocyte, neutrophil, and lymphocyte counts; the platelet-to-lymphocyte ratio; and C-reactive protein levels. We plotted receiver operating characteristic curves to determine the sensitivity and specificity of the platelet-to-lymphocyte ratio along with C-reactive protein. The areas under the curve were 0.892 for C-reactive protein and 0.707 for the platelet-to-lymphocyte ratio. For systemic inflammation, platelet-to-lymphocyte ratio can be used in conjunction with other biomarkers as an indicator of inflammation in renal transplant recipients who present with infection-associated causes to emergency departments.


[An infection model which was induced in a carboxymethyl cellulose (CMC) pouch on the back of the rat].
The air-pouch model of inflammation in rats is excellent in that it allows quantitative evaluation of inflammation, and it is used for analysis of inflammatory mediators and as an evaluation system for anti-inflammatory drugs. We investigated the possibility of using this system as an experimental infection system. As a result, inflammation was found to be caused by injection of a constant amount of Staphylococcus aureus solution (10(4)-10(8)). The amount of infiltration and the number of infiltrating cells varied with quantity of bacteria. The infiltrating cells consisted mainly of neutrophils. In this experimental model of infectious disease, the severity of inflammation could be quantitatively evaluated as a function of time in terms of bacterial proliferation and the body's response to bacterial proliferation based on the amount of fluid in the air pouch and the number of infiltrating cells, suggesting that the model is useful. In this experimental system, there were no differences between the number of live bacteria, the number of infiltrating cells or the amount of infiltration when S. aureus Smith strain and clinically-isolated methicillin-resistant S. aureus (MRSA) were used, suggesting that there is no difference between the inflammation-induced activity of MRSA and MSSA.


From virtual reality to the operating room: the endoscopic sinus surgery simulator experiment.
Establish the feasibility of a predictive validity study in sinus surgery simulation training and demonstrate the effectiveness of the Endoscopic Sinus Surgery Simulator (ES3) as a training device. Prospective, multi-institutional controlled trial. Four tertiary academic centers with accredited otolaryngology-head and neck surgery residency programs. Twelve ES3-trained novice residents were compared with 13 control novice residents. Subjects were assessed on the performance of basic sinus surgery tasks. Their first in vivo procedure was video recorded and submitted to a blinded panel of independent experts after the panel established a minimum inter-rater reliability of 80 percent. The recordings were reviewed by using a standardized computer-assisted method and customized metrics. Results were analyzed with the Mann-Whitney U test. Internal rater consistency was verified with Pearson moment correlation. Completion time was significantly shorter in the experimental group (injection P = 0.003, dissection P < 0.001), which, according to the rater panel, also demonstrated higher confidence (P = 0.009), demonstrated skill during instrument manipulation (P = 0.011), and made fewer technical mistakes during the injection task (P = 0.048) compared with the control group. The raters' post hoc internal consistency was deemed adequate (r > 0.5 between serial measurements). The validity of the ES3 as an effective surgical trainer was verified in multiple instances, including those not depending on subjective rater evaluations. The ES3 is one of the few virtual reality simulators with a comprehensive validation record. Advanced simulation technologies need more rapid implementation in otolaryngology training, as they present noteworthy potential for high-quality surgical education while meeting the necessity of patient safety.


Primary osteosarcoma of the breast.
Primary extra-osseous osteogenic sarcomas have been reported in many tissues of the body but their occurrence in the breast is rare. The tumor has poor prognosis with high risk of recurrence and hematogenous spread, most commonly to the lungs. We report a case of primary osteosarcoma of the breast occurring in a 58-year old woman. Histo-pathological examination revealed pleomorphic tumor cells with osteoid formation and positivity for immuno-histochemical stain vimentin.


Detecting blood transfusion reactions: What are vital signs anyway?
There is a lack of consistency in the scientific literature regarding what is included in vital signs and considered derangement in findings. We used vital signs during blood product administration as an exemplar to explore this controversy. Vital sign components varied across all studies when reviewed by a cohort of frontline nurses attempting to align institutional policy with current evidence. Only low-level data linking conventional approaches to vital sign monitoring for transfusion reaction detection were found.


Cisplatin, etoposide, ifosfamide, vincristine and bleomycin combination chemotherapy for far advanced testicular carcinoma.
Forty-eight patients with advanced testicular cancer, defined as abdominal mass greater than 10 cm, mediastinal mass greater than 5 cm, more than 20 lung metastases, or visceral organ involvement were treated with an intensive, alternating five-drug regimen consisting of cisplatin 50 mg/m2 d 1-3, etoposide 170 mg/m2 d 1-3, ifosfamide 5 g/m2 d 15, vincristine 2 mg weekly, bleomycin 15 mg/m2 weekly, q d 28. Thirty-four (71%) of the patients attained tumor-free status. This was achieved by chemotherapy alone in 14 patients and by surgical resection of residual disease in the remaining 20 patients (histology of resected tissue: necrosis 12, mature teratoma 7, viable carcinoma 1). Patients with pure seminoma responded better than patients with nonseminoma (CR 100% vs. 67%, respectively). In a univariate analysis only the value of HCG (less than vs greater than 10,000 U/L) and the number of involved organ sites (less than or equal to 2 vs greater than to 2) had significant influence on the response rate. After a minimum follow-up of 24 months 3 patients (9%) have relapsed. The survival rate is 76% after 36 months, with 61% remaining disease-free. Though this intensive regimen might bestow some of the therapeutic advantages of standard three-drug protocols in far advanced testicular cancer, the results are still less than optimal and warrant the exploration of new therapeutic strategies.


Properties of a Membrane-bound Phosphatase from the Thylakoids of Spinach Chloroplasts.
A 3-phosphoglycerate phosphatase activity of about 2 micromoles per minute per milligram chlorophyll is associated with the thylakoid membranes of spinach chloroplasts. The K(m) for 3-phosphoglycerate is 3 millimolar. The enzyme can be solubilized from thylakoid membranes by treatment with 0.33 molar MgCl(2) or sodium deoxycholate. The activity is not stimulated by sulfhydryl reagents or the addition of 10 millimolar MgCl(2). The enzymic activity is insensitive to ethylenediaminetetraacetate. The pH optimum is broad, between 5.5 to 7.5. Although the substrate specificity is broad, 3-phosphoglycerate is the best substrate of those tested at neutral pH. However, p-nitrophenyl phosphate was a more effective substrate at pH 5.5. The enzyme exhibits the general characteristics of an acid phosphatase.


Soft tissue body composition: familial resemblance and independent influences on bone mineral density.
Familial resemblance in fat and lean soft tissue and the influence of these body composition measures on bone mineral density (BMD) were determined in 162 adult members of 42 families. Whole body fat and lean mass and BMD of the spine and femoral neck were measured by dual-energy X-ray absorptiometry and BMD of the heel and radius by single-photon absorptiometry. Significant correlations of lean tissue were observed between mothers and daughters (r = 0.35, p < 0.05), fathers and sons (r = 0.36, p < 0.05), and daughters and sons (r - 0.42, p < 0.01) after adjustment for age, height, fat mass, physical activity, and parity (among women). Fat tissue was not significantly correlated among any family member pairs. Lean tissue, but not fat, was positively associated with BMD in one or more family member subgroups at all skeletal sites. These findings suggest a hereditary component to muscle mass. Previously documented familial similarities in bone density may be related in part to the influence of lean mass on bone density.


Foamy virus vector-mediated gene correction of a mouse model of Wiskott-Aldrich syndrome.
The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and immunodeficiency. Hematopoietic cell transplantation can cure the disease and gene therapy is being tested as an alternative treatment option. In this study, we assessed the use of foamy virus (FV) vectors as a gene transfer system for WAS, using a Was knockout (KO) mouse model. Preliminary experiments using FV vectors expressing the green fluorescent protein under the transcriptional control of the endogenous WAS promoter or a ubiquitously acting chromatin opening element allowed us to define transduction conditions resulting in high (>40%) and long-term in-vivo marking of blood cells after transplantation. In following experiments, Was KO mice were treated with FV vectors containing the human WAS complementary DNA (cDNA). Transplanted animals expressed the WAS protein (WASp) in T and B lymphocytes, as well as platelets and showed restoration of both T-cell receptor-mediated responses and B-cell migration. We also observed recovery of platelet adhesion and podosome formation in dendritic cells (DCs) of treated mice. These data demonstrate that FV vectors can be effective for hematopoietic stem cell (HSC)-directed gene correction of WAS.


[Social network analysis: a method to improve safety in healthcare organizations].
Patient safety depends on the culture of the healthcare organization involving relationships between professionals. This article proposes that the study of these relations should be conducted from a network perspective and using a methodology called Social Network Analysis (SNA). This methodology includes a set of mathematical constructs grounded in Graph Theory. With the SNA we can know aspects of the individual's position in the network (centrality) or cohesion among team members. Thus, the SNA allows to know aspects related to security such as the kind of links that can increase commitment among professionals, how to build those links, which nodes have more prestige in the team in generating confidence or collaborative network, which professionals serve as intermediaries between the subgroups of a team to transmit information or smooth conflicts, etc. Useful aspects in stablishing a safety culture. The SNA would analyze the relations among professionals, their level of communication to communicate errors and spontaneously seek help and coordination between departments to participate in projects that enhance safety. Thus, they related through a network, using the same language, a fact that helps to build a culture. In summary, we propose an approach to safety culture from a SNA perspective that would complement other commonly used methods.


Recurrent dislocation of the pisiform bone.
A 17-year-old man had chronic pain and recurrent radial dislocation of the pisiform bone following a fall on his outstretched arm. The bone was excised, along with a 3 x 4-mm osteocartilaginous fragment from the distal triquetrum. Pisotriquetral joint instability should be considered in the differential diagnosis of palmar-ulnar wrist pain.


Immune-refractory cancers and their little helpers--an extended role for immunetolerogenic MHC molecules HLA-G and HLA-E?
There is strong evidence to support a role for non-classical MHC class I (class Ib) molecules, most notably HLA-E and HLA-G in tumour immune escape. In this article, we summarize the current knowledge on their expression, regulation and functional relevance in various malignancies, particularly brain tumours. Special emphasis is devoted to the phenomenon that these tolerogenic molecules are expressed by non-transformed cells that are found in close neighborhood to tumour cells representing either parenchymal cells or immune cells attracted to the tumour microenvironment. Here they may act as "natural" or "inducible" suppressors of anti-tumoural immune responses. We thus speculate about the role of HLA-G expressing T cells, a novel population of natural regulatory cells that was identified recently. It is suggested that various cell types within a tumour cooperate in order to inhibit anti-tumour immunity-and that immunetolerogenic HLA-G may play a major role in this context.


Responding to climate change in New York State: the ClimAID integrated assessment for effective climate change adaptation in New York State. Final report.
Climate change is already beginning to affect New York State, and these impacts are projected to grow. At the same time, the state has the ability to develop adaptation strategies to prepare for and respond to climate risks now and in the future. The ClimAID assessment provides information on climate change impacts and adaptation for eight sectors in New York State: water resources, coastal zones, ecosystems, agriculture, energy, transportation,telecommunications, and public health. Observed climate trends and future climate projections were developed for seven regions across the state. Within each of the sectors, climate risks, vulnerabilities, and adaptation strategies are identified. Integrating themes across all of the sectors are equity and environmental justice and economics.Case studies are used to examine specific vulnerabilities and potential adaptation strategies in each of the eight sectors. These case studies also illustrate the linkages among climate vulnerabilities, risks, and adaptation, and demonstrate specific monitoring needs. Stakeholder participation was critical to the ClimAID assessment process to ensure relevance to decision makers across the state.


Zn(2+) rather than Ca(2+) or Mg(2+) used as a cofactor in non-muscular actin from the oyster to control protein polymerization.
The major cytoskeletal protein of most cells is actin, which polymerizes to form actin filaments (F-actin). Each actin monomer (G-actin) contains a divalent alkaline earth metal ion (in vivo Mg(2+); in vitro usually Ca(2+)) as a cofactor that is crucial for protein polymerization. Prior to this study, however, whether or not other types of metal ions can play the same role as Mg(2+) or Ca(2+) in actins remains unknown. A new actin from the gills of oyster (AGO) was prepared and characterized by protein purification techniques, SDS- and native-PAGE, and LC-MS\MS for the first time. The property of this protein was studied by CD, fluorescence and UV/vis spectroscopy, laser light scattering, and TEM. AGO is a monomer with a MW of ~42kDa. AGO is unique among all known actins in that Zn(2+) is only a naturally binding metal in the protein, and that one native AGO molecule binds 8 zinc ions, which can be removed by EDTA treatment at pH7.2. The presence of zinc has a great effect on the secondary and tertiary structure of the protein. Correlated with such effect is that these zinc ions in native AGO facilitate protein polymerization, whereas removal of zinc ions from native AGO results in a loss of such polymerization property. The present work demonstrates that AGO is a novel zinc-binding protein with high capacity, and high selectivity. This work extends an understanding of the function of zinc and actin.


Sustained-release of erythropoietin using a novel injectable thermosensitive hydrogel: in vitro studies, biological activity, and efficacy in rats.
In the current study erythropoietin (EPO) loaded trimethyl chitosan/tripolyphosphate nanoparticles-embedded in a thermosensitive hydrogel was prepared. The influence of the main experimental factors on the properties of EPO-loaded nanoparticles were evaluated using a two-factors central composite design and the optimized formulation was then freeze dried. Sodium dodecyl sulfate-page and circular dichroismspectroscopy were used to confirm the structural stability of EPO following encapsulation and freeze drying. Rheological properties, and the release rate of EPO from the hydrogel were examined. Mean particle size, zeta potential, and entrapment efficiency of the optimized EPO-loaded nanoparticles were confirmed 151.5 ± 16 nm, 11.5 ± 1.8 mV, and 78.5 ± 5.9%, respectively. The hydrogel containing nanoparticles existed as a solution at room temperature converted to a semisolid upon increasing the temperature to 35 ± 1.2 °C and demonstrated controlled release of EPO for more than 10 days. The stability of EPO in the hydrogel system was further investigated using in vivo biological activity assay and the result revealed relative potency of 0.85 as calibrated with standard EPO. Finally, a single injection of the EPO-loaded nanoparticles-embedded in the hydrogel administered to Sprague-Dawley rats resulted in elevated reticulocytes for about 20 days compared to control group received blank hydrogel.


Topology of the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (RyR1).
To define the topology of the skeletal muscle ryanodine receptor (RyR1), enhanced GFP (EGFP) was fused in-frame to the C terminus of RyR1, replacing a series of C-terminal deletions that started near the beginning or the end of predicted transmembrane helices M1-M10. The constructs were expressed in HEK-293 (human embryonic kidney cell line 293) or mouse embryonic fibroblast (MEF) cells, and confocal microscopy of intact and saponin-permeabilized cells was used to determine the subcellular location of the truncated fusion proteins. The fusion protein truncated after M3 exhibited uniform cytoplasmic fluorescence, which was lost after permeabilization, indicating that proposed M', M", M1, M2, and M3 sequences are not membrane-associated. The fusion protein truncated at the end of the M4-M5 loop and containing M4 was membrane-associated. All longer truncated fusion proteins were also associated with intracellular membranes. Mapping by protease digestion and extraction of isolated microsomes demonstrated that EGFP positioned after either M5, the N-terminal half of M7 (M7a), or M8 was located in the lumen, and that EGFP positioned after either M4, M6, the C-terminal half of M7 (M7b), or M10 was located in the cytoplasm. These results indicate that RyR1 contains eight transmembrane helices, organized as four hairpin loops. The first hairpin is likely to be made up of M4a-M4b. However, it could be made up from M3-M4, which might form a hairpin loop even though M3 alone is not membrane-associated. The other three hairpin loops are formed from M5-M6, M7a-M7b, and M8-M10. M9 is not a transmembrane helix, but it might form a selectivity filter between M8 and M10.


Regeneration of rhodopsin and bacteriorhodopsin. The role of retinal analogues as inhibitors.
The rate of regeneration of rhodopsin, from 11-cis-retinal and opsin, and bacteriorhodopsin from all-trans-retinal and bacterio-opsin, in the presence or absence of compounds whose structures partially resemble retinal were measured. Some of these compounds severely slowed down the regeneration process, but did not influence the extent of regeneration. In the case of compounds with a carbonyl functional group they were not joined to the active site of the apo-protein via a Schiff's base linkage since after treatment with NaBH4 an active apo-protein remained. The most effective inhibitors of rhodopsin regeneration were molecules whose structure could be superimposed on 9-cis or 11-cis retinal up to carbon atom 11. These C13 and C15 molecules were not distinguished between aldehyde, ketone or alcohol functional groups. The regeneration of bacteriorhodopsin was not inhibited by retinal analogues with short side chains. The most effective inhibitors were the all-trans C17-aldehyde (beta-ionylideneacetaldehyde) or C18-ketone (beta-ionylidenepent-3-ene-2-one) which, compared to retinal, lack two or three carbon atoms from the end of the poylene chain. The inhibition was very dependent upon the presence of the all-trans isomer and required aldehyde or ketone as functional group nitriles and alcohols were less effective. However, similarly to retinol, the all-trans C17 and C18 alcohols underwent a bathochromic shift and showed fine-structured spectra when mixed with bacterio-opsin.


Acute haemodynamic and proteinuric effects of prednisolone in patients with a nephrotic syndrome.
Administration of prednisolone causes an abrupt rise in proteinuria in patients with a nephrotic syndrome. To clarify the mechanisms responsible for this increase in proteinuria we have performed a placebo controlled study in 26 patients with a nephrotic syndrome. Systemic and renal haemodynamics and urinary protein excretion were measured after prednisolone and after placebo. After i.v. administration of 125-150 mg prednisolone total proteinuria increased from 6.66+/-4.42 to 9.37+/-6.07 mg/min (P<0.001). By analysing the excretion of proteins with different charge and weight (albumin, transferrin, IgG, IgG4 and beta2-microglobulin) it became apparent that the increase of proteinuria was the result of a change in size selectivity rather than a change in glomerular charge selectivity or tubular protein reabsorption. Glomerular filtration rate rose from 83+/-34 ml to 95+/-43 ml/min (P<0.001) after 5 h, whereas effective renal plasma flow and endogenous creatinine clearance remained unchanged. As a result filtration fraction was increased, compatible with an increased glomerular pressure, which probably contributes to the size selectivity changes. Since corticosteroids affect both the renin-angiotensin system and renal prostaglandins, we have evaluated the effects of prednisolone on proteinuria after pretreatment with 3 months of the angiotensin-converting enzyme inhibitor lisinopril or after 2 weeks of the prostaglandin synthesis inhibitor indomethacin. Neither drug had any effect on prednisolone-induced increases of proteinuria. Prednisolone increases proteinuria by changing the size selective barrier of the glomerular capillary. Neither the renin-angiotensin axis nor prostaglandins seem to be involved in these effects of prednisolone on proteinuria.


Species-specific features of DARC, the primate receptor for Plasmodium vivax and Plasmodium knowlesi.
The DARC (Duffy antigen/receptor for chemokines) gene, also called Duffy or FY, encodes a membrane-bound chemokine receptor. Two malaria parasites, Plasmodium vivax and Plasmodium knowlesi, use DARC to trigger internalization into red blood cells. Although much has been reported on the evolution of DARC null alleles, little is known about the evolution of the coding portion of this gene or the role that protein sequence divergence in this receptor may play in disease susceptibility or zoonosis. Here, we show that the Plasmodium interaction domain of DARC is nearly invariant in the human population, suggesting that coding polymorphism there is unlikely to play a role in differential susceptibility to infection. However, an analysis of DARC orthologs from 35 simian primate species reveals high levels of sequence divergence in the Plasmodium interaction domain. Signatures of positive selection in this domain indicate that species-specific mutations in the protein sequence of DARC could serve as barriers to the transmission of Plasmodium between primate species.


Colonic chicken skin mucosa: association with juvenile polyps in children.
Chicken skin mucosa is a newly described endoscopic finding associated with colonic neoplasms in adults. Chicken skin mucosa was sought in children with juvenile polyps to determine the prevalence, endoscopic features, and location. An alternative theory is proposed for the pathogenic mechanism of this finding. Children having colonoscopy and polypectomy were prospectively evaluated for the presence of chicken skin mucosa. The location of the polyps was determined at colonoscopy; the size of removed polyps was measured during processing of samples in pathology. Biopsies from colonic chicken skin mucosa were stained with hematoxylin and eosin and mucicarmine. Over a 1-yr period, 27 juvenile polyps were removed from 15 children at colonoscopy. Eleven of 15 children (73%) were found to have polyps with chicken skin mucosa; overall, 43% of the polyps had associated chicken skin mucosa. Chicken skin mucosa-positive polyps were larger than chicken skin mucosa-negative polyps and were only found in the rectosigmoid colon. Lipid-laden macrophages were found in all samples of chicken skin mucosa tested. Chicken skin mucosa is a common finding in children with juvenile polyps. It probably is the result of local mucosal trauma, rather than a preneoplastic lesion.


Cardiac tumors: echo assessment.
Cardiac tumors are exceedingly rare (0.001-0.03% in most autopsy series). They can be present anywhere within the heart and can be attached to any surface or be embedded in the myocardium or pericardial space. Signs and symptoms are nonspecific and highly variable related to the localization, size and composition of the cardiac mass. Echocardiography, typically performed for another indication, may be the first imaging modality alerting the clinician to the presence of a cardiac mass. Although echocardiography cannot give the histopathology, certain imaging features and adjunctive tools such as contrast imaging may aid in the differential diagnosis as do the adjunctive clinical data and the following principles: (1) thrombus or vegetations are the most likely etiology, (2) cardiac tumors are mostly secondary and (3) primary cardiac tumors are mostly benign. Although the finding of a cardiac mass on echocardiography may generate confusion, a stepwise approach may serve well practically. Herein, we will review such an approach and the role of echocardiography in the assessment of cardiac masses.


Resident satisfaction and its components in residential aged care.
The purpose of this study was to assess the direction and magnitude of the effects among the components of resident satisfaction in residential aged care and to examine if the relationships among satisfaction components vary according to facility type (i.e., nursing home and hostel). Briefly, a hostel is a low-care facility in which residents are more independent and receive personal but not nursing care. A cross-sectional survey design was adopted to collect the required information, and a stratified random sampling approach was used to select facilities. Structural equation modeling was used to examine relationships among satisfaction components in a sample of 394 nursing home and 752 hostel residents. The results indicate that satisfaction with staff care has a moderate and positive effect on all other aspects of resident satisfaction. The relationships among satisfaction components are different for nursing home and hostel residents. The findings lead to an improved understanding of the interrelationship among resident satisfaction components, which has important implications for improving quality outcomes (e.g., resident satisfaction) through appropriate intervention (e.g., enhancing staff care).


Influence of the direction of elemental luminance gradients on the responses of V4 cells to textured surfaces.
The texture of an object provides important cues for its recognition; however, little is known about the neural representation of texture. To investigate the representation of texture in the visual cortex, we recorded single-cell activities in area V4 of macaque monkeys. To distinguish the sensitivity of the cells to texture parameters such as density and element size from that to spatial frequency, we used texture stimuli mimicking shaded granular surfaces. We varied the size and density of the texture elements and the direction of elemental luminance gradients (apparent shadings) as stimulus parameters. Most macaque V4 cells (151 of 170; 89%) exhibited sensitivity to the texture parameters. Interestingly, 21 of these cells were tuned to single shading directions (unidirectional tuning). This unidirectional tuning cannot be explained by complex-cell-like tuning for spectral power of spatial frequency, because texture stimuli with a shading direction and its opposite have almost the same spectral power. Unidirectional tunings of these cells were invariant for the position of the texture elements. Thus, this tuning cannot be explained by simple-cell-like phase-dependent spatial frequency tuning or selectivity to a particular arrangement of the elements. Moreover, the unidirectional tuning had a bias toward vertical directions, consistent with an anisotropy in the perception of three-dimensional shape from shading. This novel spatial property suggests that V4 cells are involved in extracting texture features from objects, including their three-dimensionality.


Novel Classification of Acromegaly in Accordance with Immunohistochemical Subtypes: Is There Really a Clinical Relevance?
According to the recent studies, immunohistochemical subtypes of growth hormone (GH) secreting adenomas have been considered as a predictive factor in determining the clinical outcomes including biochemical, radiologic, and endocrine remission. In a 20 year-of time period, acromegaly patients who were treated and followed at the Endocrinology Department of our University Hospital were screened for the study. Of total 98 patients, 65 patients who had been operated by transsphenoidal surgery and having postoperative specimens were included. Postoperative specimens of the surgery of the patients were classified into 3 groups based on the histochemical characteristics (densely, sparsely, and mixed). Parasellar extensions of pituitary tumors were classified into the five grades according to Knosp classification. The patients were investigated and evaluated for postoperative clinical progress, remission rates, comorbidities regarding with the histopathological patterns. Of total 65 patients, 31 were classified as densely granulated (group 1), 32 were classified as sparsely granulated (group 2), and 2 patients were assessed as mixed granulated (group 3). There was no difference between groups for age and gender. Pre-treatment of adenoma size in all groups was correlated with each other and the frequency of macroadenoma (1 vs. 2, 77.4 vs. 84.3%) was higher in two groups. Although mean initial GH levels in group 1 was higher than the other groups (p=0.03), IGF1 levels (age and gender matched) were similar in each group. Adenomas in all groups demonstrated noninvasive radiological characteristics (Knosp grade 0-1-2). Ki-67 proliferation index of both groups (64.5 vs. 50%) was predominantly 1%. With a similar follow-up period, the endocrine remission rates (GH<1 μg/l) in groups were 64 vs. 69%, respectively. In conclusion, classification according to immunohistochemical subtypes of growth hormone secreting adenomas may not be a qualified parameter to evaluate patients with patterns of aggressiveness, clinical outcomes, or treatment response.


Effect of bronchial thermoplasty on structural changes and inflammatory mediators in the airways of subjects with severe asthma.
Bronchial thermoplasty (BT) is a novel technique used in the treatment of subjects with severe refractory asthma. Radiofrequency is provided to airway walls during bronchoscopy in order to reduce airway remodeling. Several clinical studies have reported an improvement in subjects' symptoms following BT. However, how BT affects the airway architectures and inflammatory mediators in the airways has not been yet fully elucidated. Fourteen subjects with severe asthma were recruited in this study according to the criteria of ATS severe asthma definition. The study subjects undertook bronchial biopsy during the bronchoscopy procedure at baseline and 6 weeks after the initial BT treatment. The obtained samples were stained with antibodies for α-smooth muscle actin (α-SMA); protein gene product (PGP) 9.5, a specific nerve marker; von Willebrand factor (vWF), a marker for blood vessels; interleukin-17A (IL-17A) and transforming growth factor-β1 (TGF-β1). The expression of α-SMA and PGP9.5 were significantly reduced post-BT. There was no significant difference in the number of blood vessels between baseline and post-BT. In addition, BT did not affect the production of IL-17A and TGF-β1 in the airways. The changes in the expression of α-SMA and PGP9.5 had no significant correlation with the improvement of pulmonary function. and Clinical Relevance: This study suggests that BT reduces airway smooth muscle mass and the airway innervation without affecting vasculature and the production of inflammatory mediators in the airways of subjects with severe asthma.


Physics and chemistry of plasma-assisted combustion.
There are several mechanisms that affect a gas when using discharge plasma to initiate combustion or to stabilize a flame. There are two thermal mechanisms-the homogeneous and inhomogeneous heating of the gas due to 'hot' atom thermalization and vibrational and electronic energy relaxation. The homogeneous heating causes the acceleration of the chemical reactions. The inhomogeneous heating generates flow perturbations, which promote increased turbulence and mixing. Non-thermal mechanisms include the ionic wind effect (the momentum transfer from an electric field to the gas due to the space charge), ion and electron drift (which can lead to additional fluxes of active radicals in the gradient flows in the electric field) and the excitation, dissociation and ionization of the gas by e-impact, which leads to non-equilibrium radical production and changes the kinetic mechanisms of ignition and combustion. These mechanisms, either together or separately, can provide additional combustion control which is necessary for ultra-lean flames, high-speed flows, cold low-pressure conditions of high-altitude gas turbine engine relight, detonation initiation in pulsed detonation engines and distributed ignition control in homogeneous charge-compression ignition engines, among others. Despite the lack of knowledge in mechanism details, non-equilibrium plasma demonstrates great potential for controlling ultra-lean, ultra-fast, low-temperature flames and is extremely promising technology for a very wide range of applications.


Ultrasensitive and Portable Assay for Lead(II) Ions by Electronic Balance as a Readout.
Lead ions (Pb2+) cause harm to human health. Therefore, the development of fast, effective, and convenient sensors for Pb2+ monitoring has received great attention. In this study, a portable method has been proposed for Pb2+ detection using normal electronic balance as a readout. Magnetic bead-catalytic strand is hybridized with platinum nanoparticles (Pt NPs) functioned substrate strand (Pt-Sub) to form double-stranded DNA first. In the presence of Pb2+, the DNAzyme is activated and cleaved at the ribo-adenosine site of the substrate strand and hence causes Pt NPs to be released into the supernatant, which can be easily separated from the Pt-Sub by a magnet. The separated Pt NPs can effectively catalyze the decomposition of H2O2 to produce O2. In a sealed bottle, the pressure inside the bottle is increased by the generation of oxygen so that the water is discharged from the drainage device, and the weight of the water can be easily and precisely measured by a normal electronic balance. The weighting water has a linear relationship with the concentration of Pb2+ in the range of 2.5-100 nM and the detection limit of 0.83 nM (S/N = 3). The proposed method has been applied to detect Pb2+ in water with satisfactory results. Because the electronic balance is one of the most commonly used analytical tools for the laboratory, it is very practical and convenient without the need for expensive instruments and complicated data processing.


From diminished men to conditionally masculine: sexuality and Australian men and adolescent boys with intellectual disability.
Men and boys with intellectual disability represent a unique group who have hitherto been overlooked by researchers and theorists exploring men and masculinities. Qualitative data from an Australian ethnographic study focused on the sexual health needs of men and adolescent boys with moderate to profound intellectual disability. Findings suggest that masculinity for this group of men is more a biopsychosocial phenomenon than a social construct organised around heteronormative ideals. The conditional masculinity of the men participating in the study was based instead on a number of intrinsic and external factors, which are described in detail.


Controlled aqueous polymerization of acrylamides and acrylates and "in situ" depolymerization in the presence of dissolved CO2.
Aqueous copper-mediated radical polymerization of acrylamides and acrylates in carbonated water resulted in high monomer conversions (t < 10 min) before undergoing depolymerization (60 min > t > 10 min). The regenerated monomer was characterized and repolymerized following deoxygenation of the resulting solutions to reyield polymers in high conversions that exhibit low dispersities.


Drug treatment of type 2 diabetes in adults.
This article outlines the pharmacological management of type 2 diabetes and its complications in adults. Practical aspects of the drug options and the evidence supporting their use are discussed. The need for a holistic, patient-centred approach to achieve the treatment goals of glycaemic control and prevention of complications is emphasised.


A Prospective Study of Overuse Knee Injuries Among Female Athletes With Muscle Imbalances and Structural Abnormalities.
OBJECTIVE: To prospectively examine the influence of hamstring-to-quadriceps (H:Q) ratio and structural abnormalities on the prevalence of overuse knee injuries among female collegiate athletes. DESIGN AND SETTING: We used chi-square 2 x 2 contingency tables and the Fischer exact test to examine associations among H:Q ratios, structural abnormalities, and overuse knee injuries. SUBJECTS: Fifty-three apparently healthy women (age = 19.4 +/- 1.3 years, height = 167.6 +/- 10.1 cm, mass = 65.0 +/- 10.0 kg) from National Collegiate Athletic Association Division I women's field hockey (n = 23), soccer (n = 20), and basketball teams (n = 10) volunteered. MEASUREMENTS: The H:Q ratio was determined from a preseason isokinetic test on a Biodex system at 60 degrees /s and 300 degrees /s. We measured athletes for genu recurvatum and Q-angles with a 14-in (35.56-cm) goniometer. Iliotibial band flexibility was assessed via the Ober test. RESULTS: Ten overuse knee injuries (iliotibial band friction syndromes = 5, patellar tendinitis = 3, patellofemoral syndrome = 1, pes anserine tendinitis = 1) occurred in 9 athletes. The H:Q ratio below the normal range at 300 degrees /s (P = 0.047) was associated with overuse knee injuries, as was the presence of genu recurvatum (P = 0.004). In addition, athletes possessing lower H:Q ratios at 300 degrees /s and genu recurvatum incurred more overuse knee injuries than athletes without these abnormalities (P = 0.001). CONCLUSIONS: The presence of genu recurvatum and an H: Q ratio below normal range was associated with an increased prevalence of overuse knee injuries among female collegiate athletes. Further investigation is needed to clarify which preseason screening procedures may identify collegiate athletes who are susceptible to overuse knee injuries.


Immune system-related CD9 is expressed in mouse central nervous system myelin at a very late stage of myelination.
CD9 is a tetra-membrane-spanning glycoprotein involved in cell adhesion, migration, and proliferation in both the immune and the immature nervous system. In this study, CD9 expression was detected in myelin of mouse brain, starting at postnatal day 16. The amount of CD9 protein continuously increased with age and persisted in the adult brain. It appeared later than myelin proteolipid protein (PLP), a typical late myelin marker. Mature oligodendrocytes were abundant in CD9, although it was also detected in astrocytes and microglial cells in vitro. CD9 appeared at the end of the myelination process and was localized along the outermost membrane of compact myelin. CD9 is known to associate with integrins, which are candidate receptors for extracellular matrix and transmit extracellular signals into the cells. Taken together, CD9 at the surface of central nervous system (CNS) mature myelin may have a unique function to facilitate signal transduction and enhance myelin membrane adhesion to extracellular matrices at very late stages of development.


Genome profiles in maternal blood during early onset preeclampsia and towards term.
inflammatory processes are present during preeclampsia and in normal pregnancy. Maternal inflammatory reactions may change towards term. Our objective was to evaluate genome signaling in blood during preeclampsia and towards term using microarrays. RNA microarrays (Illumina) were conducted on blood from preeclamptic pregnancies delivered preterm, normal pregnancies at term and normal pregnancies at gestational week 31. Two statistical methods (Q-value cut-off 1%) identified data structures in the three groups and retrieved activated genes along a time axis and a diseased-healthy axis. Signaling genes were localized within known pathways and gene sets, and genes associated with inflammation were identified. early onset preeclampsia and term pregnancies both showed distinct expression patterns when compared to normal pregnancy at gestational week 31. In preeclampsia, 19 genes were differentially expressed, including a down-regulation of CC-chemokine receptor 3 (CCR3). Among the 183 differentially expressed genes towards term, tumor necrosis factor superfamily member 15 (TNFSF15) was up-regulated and interferon-γ receptor 2 (IFNGR2) and CXC-chemokine receptor type 4 (CXCR4) were down-regulated. Seven of the genes were similarly changed during preeclampsia and towards term. a possible type 1 immune response was identified both during preeclampsia and towards term. In pre-eclampsia a premature activation of leucocytes might be present.


Effect of the beta-adrenergic agonist cimaterol (CL 263,780) on the growth and carcass characteristics of finishing swine.
A randomized complete-block design was used to evaluate the effects of the beta-adrenergic agonist, cimaterol (CL 263,780), on growth rate, feed efficiency and carcass composition of finishing swine. The drug was fed at four levels (0, .25, .5 and 1.0 ppm) to a total of 240 pigs from 64.5 to 103.7 kg live weight. Growth rate and feed efficiency were measured during the 7-wk feeding trial. Feeding cimaterol depressed feed intake, improved feed efficiency and did not alter rate of gain. Carcass evaluation showed that pigs continuously fed cimaterol had 13.2, 9.3 and 9.2% less fat measured at the 10th rib, P2 and average backfat (BF) locations, respectively, compared with controls. Cimaterol-fed pigs had increased loin eye areas (10.9%), and increased semitendinosus (11.8%) and biceps femoris (8.9%) weights compared with controls. The semitendinosus muscles of the cimaterol-fed pigs had less fat and the femur bones were shorter and lighter weight than controls. There were no detected differences in structural soundness of the live pigs, but postmortem evaluation of the hooves indicated that pigs fed 1.0 ppm cimaterol had a higher incidence of hoof lesions. Pigs withdrawn from cimaterol for 7 d were comparable in performance and carcass characteristics with those continuously fed the drug except that carcass fat measurements had generally returned to control values. The data indicate that cimaterol improved the feed efficiency of finishing pigs and increased the lean:fat ratio of their carcasses. Withdrawal of cimaterol caused compensatory fat deposition.


Glucuronidation of thyroid hormone in rat liver: effects of in vivo treatment with microsomal enzyme inducers and in vitro assay conditions.
We investigated the effects of in vivo treatment with different microsomal enzyme inducers, including clofibrate (CLOF), hexachlorobenzene (HCB), 3-methylcholanthrene (MC), 3,3',4,4'-tetrachlorobiphenyl (TCB), and 2,3,7,8-tetrachloro-p-dioxin, as well as of in vitro addition of the detergent Brij 56 on the glucuronidation of T4, T3, and rT3 by UDP-glucuronyltransferase (UGT) activities of rat liver microsomes. The results were compared with measurements of UGT activities for bilirubin, p-nitrophenol (PNP), and androsterone. In general, glucuronidation rates were 5-fold or more higher with rT3 than with T4 or T3 as substrate. In liver microsomes from untreated rats, T4 UGT activity was stimulated by Brij 56 to a maximum of about 2-fold at 0.025% detergent. Treatment of Wistar rats for 4 days with CLOF (200 mg/kg BW.day) resulted in significant increases in UGT activities for T4 (to 154%), rT3 (to 155%), and bilirubin (to 194%), in particular if assayed in the presence of 0.025% Brij 56, but had little effect on the UGT activities for T3, PNP, and androsterone. The CLOF-induced increases in T4 and rT3 UGT activities were not observed in Gunn rats, which have a complete lack of bilirubin UGT activity and greatly impaired PNP UGT activity. Treatment of Wistar rats with a single injection of MC (50 mg/kg BW), TCB (50 mg/kg BW), or 2,3,7,8-tetrachloro-p-dioxin (6.25 micrograms/kg BW) resulted, after 4 days, in 6.3- to 7.3-fold increases in T4 UGT activity and 15.1- to 16.7-fold increases in rT3 UGT activity if determined in the absence of Brij 56, whereas T4 UGT activity was only increased by 33-68% when assayed in the presence of Brij 56. T3 glucuronidation was not affected (with Brij 56) or was increased by only 33-68% (without Brij 56) after treatment with these MC-type inducers. PNP UGT activity was induced 3.6- to 4.3-fold, whereas bilirubin and androsterone UGT activities were changed little by these treatments. Similar findings regarding T4, rT3, PNP, and bilirubin UGT activities were obtained after chronic treatment of WAG rats with HCB, another MC-type inducer. However, WAG rats lack androsterone UGT and show low T3 UGT activity, which was increased about 2.3-fold by HCB treatment. On the basis of these and previous findings it is concluded that at least three UGT isoenzymes are involved in the glucuronidation of thyroid hormone.(ABSTRACT TRUNCATED AT 400 WORDS)


[Molecular factors determine primary and secondary therapy of breast carcinoma].
The clinical oncology realizes that the classical approach with systemic adjuvant chemo- and hormonal therapies is not sufficient and will be challenged by cellular and molecular structures which reflect the targets for new therapeutic approaches. These targets are key proteins involved in the signal transduction cascade. In human tumors these proteins have either lost their biological functionality by oncogenic mutations or are constitutively activated. The molecular classification of primary breast cancer was performed by assessing the following factors: estrogen- and progesterone receptors, ERbB-2 mutated p53, uPA, PAI-I, VEGF, DNA-Index and S-Phase. These factors are of prognostic and predictive value.


Purification and properties of liver arginase from teleostean fish Clarias batrachus (L.).
Liver arginase of Clarias batrachus has been purified to 56.3-fold employing ammonium sulphate fraction, DEAE-cellulose and CM-cellulose chromatography. Bidirectional polyacrylamide gel electrophoresis shows the presence of two isoenzymes of arginase. The enzyme has a molecular weight of about 87,000 and Km 15.38 mM for L-arginine, optimum pH 9.5 and temperature 37 degrees C. Ornithine and leucine as competitive whereas valine and isoleucine act as non-competitive inhibitors with respect to L-arginine as substrate.


The effect of glucagon-like peptide 2 injection on performance, small intestinal morphology, and nutrient transporter expression of stressed broiler chickens.
An experiment was conducted to determine the effect of injecting glucagon-like peptide 2 (GLP-2) on the small intestinal weight, morphology, and nutrient transporter expression in pharmacologically stressed broiler chickens. A total of 144 seven-day-old birds were fed either a basal diet (CTRL) or a basal diet plus 30 mg of corticosterone (CORT)/kg of diet for a total of 14 d. Half of the birds from each group were injected daily with GLP-2 (6.7 nmol/kg of BW) or saline for 14 d. The average final BW, ADG, ADFI, and the ratio of feed intake to weight gain (F:G) was recorded over 21 d for the 4 groups of 36 birds, namely CTRL + saline, CTRL + GLP-2, CORT + saline, and CORT + GLP-2. In addition, the absolute and relative small intestinal weight, villus height (VH), and crypt depth (CD) of the duodenum and jejunum, as well as the abundance of sodium and glucose co-transporter 1 (SGLT-1), vitamin D-dependent calcium-binding protein-28,000 molecular weight (CaBP-D28k), and peptide transporter 1 (PepT-1) mRNA in the duodenum and of liver fatty acid-binding protein (L-FABP) mRNA in the jejunum. The total DNA, RNA, and protein content in small intestinal mucosa were also determined. The results showed that CORT administration significantly lowered average final BW, ADG, ADFI, absolute small intestinal weight, VH, and CD of duodenum and jejunum (P < 0.05) while increasing the relative small intestinal weight, F:G, relative abundance of SGLT-1, CaBP-D28k, PepT-1, and L-FABP mRNA (P < 0.05). Glucagon-like peptide 2 injection increased the average final BW, ADG, VH, and CD in duodenum and jejunum and relative abundance of SGLT-1, CaBP-28k, PepT1, and PepT1 mRNA of broiler chickens, respectively (P < 0.05), and decreased F:G (P < 0.05). In chickens fed basal diet plus CORT, injecting GLP-2 decreased F:G (P < 0.05); increased VH and CD of duodenum and CD of jejunum; and increased relative abundance of SGLT-1, CaBP-D28k, PepT-1, and L-FABP mRNA, RNA, and total protein content in small intestine compared with the injection of saline (P < 0.05). In birds fed the basal diet, GLP-2 injection decreased F:G (P < 0.05) and increased final BW, ADG, small bowel weight, CD of jejunum, and relative abundance of CaBP-D28k and PepT-1 mRNA compared with injecting saline (P < 0.05). In conclusion, GLP-2 injection reversed the negative effect of stress on the weight and morphology and the absorptive function of small bowel of broiler chickens. Glucagon-like peptide 2 injection also had a positive effect on the growth performance of healthy broiler chickens.


Can Parenting Microprotections Buffer Against Adolescents' Experiences of Racial Discrimination?
Previous research highlights the importance and pervasiveness of racial discrimination for minority youth in the United States. Adolescents may experience either personal or group race-based discrimination. While past research found both forms of discrimination are harmful to well-being it is unknown whether parental microprotections, which may buffer against the negative effects of discrimination, protect against both forms of discrimination. Informed by ecological frameworks the present study examined whether parent microprotections (parental warmth/acceptance, cultural socialization, preparation for bias) buffered the effects of personal and group discrimination on adolescents' depressive symptoms. Participants were African American early adolescents (N = 129; 58% female) and their parents (90% mothers). Adolescents attended a Midwestern, Title 1, urban, public middle school and completed surveys in their homerooms and parents completed paper-pencil surveys at home, online or surveys via telephone. Multiple regression analyses were conducted to test whether adolescents' personal and group discrimination at school were related to their reports of depressive symptoms and evaluate whether parent microprotections buffered these associations. Results showed that parental microprotections moderated the effects of personal discrimination on depressive symptoms but did not buffer the effects of group discrimination. These results highlight the importance of distinguishing between personal and group discrimination. Further, additional protective factors need to be identified and tested to better understand parental actions that potentially buffer the negative effects of discrimination.


Self- and manual mobilization improves spine mobility in men with ankylosing spondylitis--a randomized study.
To evaluate effects of physiotherapeutic intervention in terms of self- and manual mobilization on chest expansion, vital capacity, posture, spine mobility and experienced consequences of the disease in patients with ankylosing spondylitis. A prospective, randomized controlled study. Thirty-two men, aged between 23 and 60 years, with ankylosing spondylitis were randomized to active or no treatment for eight weeks. Physiotherapeutic intervention included individualized self- and manual mobilization for 1 hour twice a week and individually adjusted home exercises. Two blinded investigators made the assessments of chest expansion, posture and spinal mobility before and after the treatment period. The patient filled in three and the physiotherapist one of the four Bath Ankylosing Spondylitis scales (BAS scales). In the treatment group chest expansion increased at the level of processus xiphoideus (P<0.01), with no difference in vital capacity compared with the control group. The posture improved in the cervical (C7-wall distance) (P<0.001) and in the thoracic spine (P<0.05). Thoracic and lumbar spine flexion improved (P<0.01) as did sagittal range of motion P<0.001 and P<0.01, respectively. The Bath Ankylosing Spondylitis Metrology Index total scoring improved (P<0.001) in the treatment group compared with the control group. The other three BAS scales showed no differences between groups. At four months follow-up of the treatment group, cervical spine posture, lumbar flexion and range of motion as well as BAS Metrology Index were still improved. This study shows that eight weeks of self- and manual mobilization treatment improved chest expansion, posture and spine mobility in patients with ankylosing spondylitis.


Native soluble CD5 delivers a costimulatory signal to resting human B lymphocytes.
Recently, we reported that the CD5 protein can bind to the B cell antigen CD72. So far, no functional evidence has been given for this interaction. We used soluble native CD5 and two anti-CD72 monoclonal antibodies, JT3 and WL225, produced and characterized in our laboratory in order to investigate the role of CD5 in B cell activation. Neither the CD5 nor the antibodies induced thymidine incorporation when added to resting human B cells, but they produced a two- to five-fold increase in thymidine uptake of B cells activated using immobilized anti-sIgM mAb when compared to the cultures stimulated by anti-sIgM mAb alone. The CD5 protein was effective at concentrations as low as 0.15 microM and its effect could be abolished by preincubation with soluble recombinant CD72 but not by preincubation with control proteins, indicating the specificity of the binding. The two antibodies but not the soluble CD5 produced a costimulatory effect when B cells were stimulated with suboptimal concentrations of anti-CD40 mAb or IL-4. Altogether these data suggest that a costimulatory signal can be delivered to human B cells by CD5/CD72 interaction. The possible role of CD5/CD72 signalling in physiologic humoral responses is discussed here.


Create Solidarity Networks: Dialogs in Reddit to Overcome Depression and Suicidal Ideation among Males.
The emerging scientific literature examines masculinity and gender roles as risk factors for suicide ideation or suicide in young adults and adolescents. In this vein, recent studies show that certain traditional masculine norms are related to poorer mental health-related outcomes, which influences suicide and suicide ideation. This study contributes with new understandings about the associations between masculinity and suicidal ideation among males through Reddit debates in English. The posts with more interactions referring to masculinity in the topics gender and education have been selected on Reddit, emphasizing transformative personal experiences potentially helping avoid suicide ideation. Through the analysis of Reddit posts, it is shown how users can generate spaces to express the diverse ways to live with masculinity. The discussions on Reddit in the different areas selected demonstrate the existence of proposals on how to overcome fears and facilitate relaxation of norms regarding self-reliance to encourage help-seeking when feeling depressed and therefore at greater risk of suicide ideation. The results highlight the potential importance of platforms such as Reddit to create solidarity networks, showing multiple ways of being a man and demystifying dominant masculinity by sharing different experiences.


Cone-beam dental computerized tomography for evaluating changes of aging in the dimensions central superior incisor root canals.
Cone-beam dental computerize tomography (CBCT), a noninvasive imaging method, was used to image the pulp-dentin complex. Pulp changes histologically with aging. While many studies have investigated the histological changes in pulp, few studies have focused on the changes in the shape of the root canals. This study evaluated the changes in the root canals with aging of central superior teeth using CBCT. The study examined 100 non-carious maxillary central teeth. These teeth were divided into five groups according to the age of the patients: Group A:15-24, Group B: 25-34, Group C: 35-44, Group D: 45-54 and Group E: 55 years and older. CBCT was used to determine root length, and pulp width at the cervical, apical 1/2, and apical 1/3. On comparing the groups using one-way analysis of variance (ANOVA), the root length did not differ (P>0.05), while the pulp width at the cervical, apical 1/2, and apical 1/3 differed between the groups (P<0.001). The pulp length did not change with aging, while the pulp width did change. CBCT can be used to determine the precise root length and width, to prevent iatrogenic exposure of the apex, complementing existing methods. This will improve the prediction of the prognosis of root canal treatment.


Conventional and Regulatory CD4+ T Cells That Share Identical TCRs Are Derived from Common Clones.
Results from studies comparing the diversity and specificity of the TCR repertoires expressed by conventional (Tconv) and regulatory (Treg) CD4+ T cell have varied depending on the experimental system employed. We developed a new model in which T cells express a single fixed TCRα chain, randomly rearranged endogenous TCRβ chains, and a Foxp3-GFP reporter. We purified CD4+Foxp3- and CD4+Foxp3+ cells, then performed biased controlled multiplex PCR and high throughput sequencing of endogenous TCRβ chains. We identified >7,000 different TCRβ sequences in the periphery of 5 individual mice. On average, ~12% of TCR sequences were expressed by both conventional and regulatory populations within individual mice. The CD4+ T cells that expressed shared TCR sequences were present at higher frequencies compared to T cells expressing non-shared TCRs. Furthermore, nearly all (>90%) of the TCR sequences that were shared within mice were identical at the DNA sequence level, indicating that conventional and regulatory T cells that express shared TCRs are derived from common clones. Analysis of TCR repertoire overlap in the thymus reveals that a large proportion of Tconv and Treg sharing observed in the periphery is due to clonal expansion in the thymus. Together these data show that there are a limited number of TCR sequences shared between Tconv and Tregs. Also, Tconv and Tregs sharing identical TCRs are found at relatively high frequencies and are derived from common progenitors, of which a large portion are generated in the thymus.


Survival and complications in patients with haemoglobin E thalassaemia in Sri Lanka: a prospective, longitudinal cohort study.
Worldwide, haemoglobin E β-thalassaemia is the most common genotype of severe β-thalassaemia. The paucity of long-term data for this form of thalassaemia makes evidence-based management challenging. We did a long-term observational study to define factors associated with survival and complications in patients with haemoglobin E thalassaemia. In this prospective, longitudinal cohort study, we included all patients with haemoglobin E thalassaemia who attended the National Thalassaemia Centre in Kurunegala, Sri Lanka, between Jan 1, 1997, and Dec 31, 2001. Patients were assessed up to three times a year. Approaches to blood transfusions, splenectomy, and chelation therapy shifted during this period. Survival rates between groups were evaluated using Kaplan-Meier survival function estimate curves and Cox proportional hazards models were used to identify risk factors for mortality. 109 patients (54 [50%] male; 55 [50%] female) were recruited and followed up for a median of 18 years (IQR 14-20). Median age at recruitment was 13 years (range 8-21). 32 (29%) patients died during follow-up. Median survival in all patients was 49 years (95% CI 45-not reached). Median survival was worse among male patients (hazard ratio [HR] 2·51, 95% CI 1·16-5·43), patients with a history of serious infections (adjusted HR 8·49, 2·90-24·84), and those with higher estimated body iron burdens as estimated by serum ferritin concentration (adjusted HR 1·03, 1·01-1·06 per 100 units). Splenectomy, while not associated with statistically significant increases in the risks of death or serious infections, ultimately did not eliminate a requirement for scheduled transfusions in 42 (58%) of 73 patients. Haemoglobin concentration less than or equal to 4·5 g/dL (vs concentration >4·5 g/dL), serum ferritin concentration more than 1300 μg/L (vs concentration ≤1300 μg/L), and liver iron concentration more than 5 mg/g dry weight of liver (vs concentration ≤5 mg/g) were associated with poorer survival. Patients with haemoglobin E thalassaemia often had complications and shortened survival compared with that reported in high-resource countries for thalassaemia major and for thalassaemia intermedia not involving an allele for haemoglobin E. Approaches to management in this disorder remain uncertain and prospective studies should evaluate if altered transfusion regimens, with improved control of body iron, can improve survival. Wellcome Trust, Medical Research Council, US March of Dimes, Anthony Cerami and Ann Dunne Foundation for World Health, and Hemoglobal.


Stability related bias in residues replacing glycines within the collagen triple helix (Gly-Xaa-Yaa) in inherited connective tissue disorders.
A missense mutation leading to the replacement of one Gly in the (Gly-Xaa-Yaa)n repeat of the collagen triple helix can cause a range of heritable connective tissue disorders that depend on the gene in which the mutation occurs. Osteogenesis imperfecta results from mutations in type I collagen, Ehlers-Danlos syndrome type IV from mutations in type III collagen, Alport syndrome from mutations in type IV collagen, and dystrophic epidermolysis bullosa from mutations in type VII collagen. The predicted rates of substitutions by different amino acids for glycine in the alpha1(I), alpha2(I), alpha1(III), alpha5(IV), and alpha1(VII) chains (encoded by COL1A1, COL1A2, COL3A1, COL4A5, and COL7A1, respectively) were compared with missense mutations in those chains that have been observed to cause disease. The spectrum of amino acids replacing Gly was not significantly different from that expected for the alpha1(VII) chains, suggesting that any Gly replacement will cause disease. The distribution of residues replacing Gly was significantly different from that expected for all other collagen chains studied, with a particularly strong bias seen for alpha1(I) and alpha1(III) collagen chains. The bias did not correlate with the degree of chemical dissimilarity between Gly and the replacement residues, but in some cases a relationship was observed with the predicted extent of destabilization of the triple helix. For alpha1(III) collagen chains, the more destabilizing mutations were identified more often than expected. For alpha1(I), the most destabilizing residues, Val, Glu, and Asp, and the least destabilizing residue, Ala, were underrepresented. This bias supports the hypothesis that the level of triple-helix destabilization determines clinical outcome.


Implementation of patient dosimetry in the clinical practice after targeted radiotherapy using [177Lu-[DOTA0, Tyr3]-octreotate.
This study's aim was to develop our dosimetric methodology using a commercial workstation for the routine evaluation of the organs at risk during peptide receptor radionuclide therapy (PRRT) with 177Lu. First, planar and SPECT sensitivity factors were determined on phantoms. The reconstruction parameters were optimized by SPECT/CT image acquisition using a NEMA IEC phantom containing a 500 ml bottle of 177Lu, to simulate a kidney. The recovery coefficients were determined on various phantoms. For the red marrow, this was calculated using a NEMA IEC phantom that contained a centrally placed bottle of 80 ml of 177Lu (to model the L2-L4 red marrow) flanked by two 200 ml bottles with 177Lu to simulate the kidneys. Then, SPECT/CT images were acquired at 4, 24, 72, and 192 h after injection in 12 patients with neuroendocrine tumors who underwent PRRT with 177Lu-DOTATATE. SPECT data were reconstructed using the iterative ordered subset expectation maximization (OSEM) method, with six iterations and ten subsets, attenuation, scatter, recovery resolution corrections, and a Gaussian post-filter of 0.11 cm. The liver, spleen, kidneys, and red marrow dose per administered activity (AD/A admin) values were calculated with the Medical Internal Radiation Dose (MIRD) formalism and the residence times (Dosimetry toolkit® application) using standard and CT imaging-based organ masses (OLINDA/EXM® V1.0 software). Sensitivity factors of 6.11 ± 0.01 and 5.67 ± 0.08 counts/s/MBq were obtained with planar and SPECT/CT acquisitions, respectively. A recovery coefficient of 0.78 was obtained for the modeled L2-L4 red marrow. The mean AD/A admin values were 0.43 ± 0.13 mGy/MBq [0.27-0.91] for kidneys, 0.54 ± 0.58 mGy/MBq [0.12-2.26] for liver, 0.61 ± 0.13 mGy/MBq [0.42-0.89] for spleen, and 0.04 ± 0.02 mGy/MBq [0.01-0.09] for red marrow. The AD/A admin values varied when calculated using the personalized and standard organ mass, particularly for kidneys (p = 1 × 10-7), spleen (p = 0.0069), and red marrow (p = 0.0027). Intra-patient differences were observed especially in organs close to or including tumor cells or metastases. The obtained AD/A admin values were in agreement with the literature data. This study shows the technical feasibility of patient dosimetry in clinical practice and the need to obtain patient-specific information.


Do Student Characteristics Affect Teachers' Decisions to Use 1:1 Instruction?
One-to-one instruction is a critical component of evidence-based instruction for students with autism spectrum disorder, but is not used as often as recommended. Student characteristics may affect teachers' decisions to select a treatment and/or implement it. This study examined the associations between students' clinical and demographic characteristics and teachers' reported use of discrete trial training (DTT) and pivotal response training (PRT). Children's higher sensory symptoms, lower social approach, lower verbal skills and higher self-regulation difficulties were associated with more frequent 1:1 DTT and PRT. Results suggest that teachers give more frequent 1:1 instruction to children with more observable impairments, do not match children to type of 1:1 intervention, and may inadvertently neglect other students for whom individualized intervention may still be beneficial.


A metagenomics method for the quantitative detection of bacterial pathogens causing hospital-associated and ventilator-associated pneumonia.
The management of ventilator-associated pneumonia and hospital-acquired pneumonia requires rapid and accurate quantitative detection of the infecting pathogen. To this end, we propose a metagenomic sequencing assay that includes the use of an internal sample processing control for the quantitative detection of 20 relevant bacterial species from bronchoalveolar lavage samples.