Molecular docking and molecular dynamics simulation studies of Trypanosoma cruzi triosephosphate isomerase inhibitors. Insights into the inhibition mechanism and selectivity.
Trypanosoma cruzi (T. cruzi) triosephosphate isomerase (TcTIM) is a glycolytic enzyme essential for parasite survival and has been considered an interesting target for the development of new antichagasic compounds. The homodimeric enzyme is catalytically active only as a dimer. Interestingly, significant differences exist between the human and parasite TIMs interfaces with a sequence identity of 52%. Therefore, compounds able to specifically disrupt TcTIM but not Homo sapiens TIM (hTIM) dimer interface could become selective antichagasic drugs. In the present work, the binding modes of 1,2,4-thiadiazol, phenazine and 1,2,6-thiadiazine derivatives to TcTIM were investigated using molecular docking combined with molecular dynamics (MD) simulations. The results show that phenazine and 1,2,6-thiadiazine derivatives, 2 and 3, act as dimer-disrupting inhibitors of TcTIM having also allosteric effects in the conformation of the active site. On the other hand, the 1,2,4-thiadiazol derivative 1 binds into the active site causing a significant decrease in enzyme mobility in both monomers. The loss of conformational flexibility upon compound 1 binding suggests that this inhibitor could be preventing essential motions of the enzyme required for optimal activity. The lack of inhibitory activity of 1 against hTIM was also investigated and seems to be related with the high mobility of hTIM which would hinder the formation of a stable ligand-enzyme complex. This work has contributed to understand the mechanism of action of this kind of inhibitors and could result of great help for future rational novel drug design.


Sympathomimetic actions of reserpine administered during treatment with dopamine in the dog.
Reserpine injected intravenously during infusion with dopamine brought about sympathomimetic effects, but a second injection of reserpine after 120 minutes did not elicit such effects. This pressor effect was eliminated by phenoxybenzamine and this positive chronotrpic effect by propranolol. Reserpine induced similar but weak sympathomimetic effects after cocaine, while it induced no changes in blood pressure and heart rate after infusion with noradrenaline. Meanwhile, pressor response to dopamine was potentiated 24 hours after reserpine and was further potentiated after additional infusion with noradrenaline. The sympathomimetic actions induced by the concurrent administration of reserpine and dopamine may be attributable to a facilitation of those indirect mechanisms, prinicpally of endogenous catecholamine release.


Time-dependent bioaccumulation of distinct rod-type TiO2 nanoparticles: comparison by crystalline phase.
A complete understanding of the interaction between nanoparticles and biological systems, including nanoparticle uptake and distribution and the biological responses, could guide the design of safer and more effective nanoparticles than those currently available. In this study, we compared the distribution in mice over time of two rod-type titanium dioxide nanoparticles (TiNPs) that feature distinct phases, anatase (ATO) and brookite (BTO). Surface areas of BTO and ATO were estimated to be 102 and 268 m(2) g(-1) , respectively, and negative charge on the surface of ATO was higher than that of BTO in deionized water. Both TiNPs were rapidly distributed into tissues after injection. At 4 weeks after injection, both TiNPs were maximally accumulated in the spleen, followed by the liver, but the total accumulation of ATO in tissues measured in this study was more than that of BTO. Moreover, the cellular antioxidant function was similar although the levels of Ti measured in tissues were distinct between the two TiNPs. Based on these results, we suggest that the fate of TiNPs in the body may differ according to the size and surface charge of the TiNPs even when their shape is the same.


A Bayesian autoregressive three-state hidden Markov model for identifying switching monotonic regimes in microarray time course data.
When modeling time course microarray data special interest may reside in identifying time frames in which gene expression levels follow a monotonic (increasing or decreasing) trend. A trajectory may change its regime because of the reaction to treatment or of a natural developmental phase, as in our motivating example about identification of genes involved in embryo development of mice with the 22q11 deletion. To this aim we propose a new flexible Bayesian autoregressive hidden Markov model based on three latent states, corresponding to stationarity, to an increasing and to a decreasing trend. In order to select a list of genes, we propose decision criteria based on the posterior distribution of the parameters of interest, taking into account the uncertainty in parameter estimates. We also compare the proposed model with two simpler models based on constrained formulations of the probability transition matrix.


Recombinant bovine somatotropin blunts plasma tumor necrosis factor-alpha, cortisol, and thromboxane-B2 responses to endotoxin in vivo.
In vivo studies determined the effects of recombinant bovine somatotropin (bST; sometribove) administration (0.1 mg/kg.day, im) to calves on the increases in plasma immunoreactive tumor necrosis factor-alpha (TNF alpha), prostacyclin [6-keto-prostaglandin F1 alpha (6KP)], thromboxane-B2 (TXB), and cortisol (C) that occurred after endotoxin challenge (ET). Two ETs were administered 5 days apart to test the effect of bST on the natural attenuation of hormone and cytokine responses that occurs after repeated challenge with endotoxin. Calves (n = 6) were treated with bST for 5 days. On day 6, the first ET was administered (Escherichia coli; 055:B5; 0.2 microgram/kg, i.v.). Blood was sampled before and hourly after ET through 6 h. For the next 4 days, bST injections continued, and ET was repeated 1 day later. Six additional calves were treated with bicarbonate buffer as contemporary controls for the bST and were similarly challenged with endotoxin. Plasma TNF alpha, C, 6KP, and TXB were significantly increased after each ET. The increases in TNF alpha, C, and TXB were blunted after the second Et compared to those after the first in both untreated and bST-treated animals. The increases in plasma TNF alpha and C and peak plasma TXB and TXB/6KP ratios were smaller in bST-treated than in untreated after the first ET. TNF alpha receptor binding was studied in hepatic microsomal fractions from three bST-treated and three untreated calves. Microsomal fractions from bST-treated calves bound 40% less TNF alpha (5.97 vs. 9.96 pmol/mg) than similar fractions from controls. The data indicate that bST decreases TNF alpha, TXB, and C responses to ET and reduces the TNF alpha-binding capacity of hepatic membranes, suggesting a multiplicity of sites where bST might affect the physiological response to endotoxin.


Fine-needle aspiration cytology of metastatic carcinoid tumor: report of a case and review of the literature.
While carcinoid tumor is a relatively common neoplasm in surgical pathology, fine-needle aspiration (FNA) cytology as a method of primary diagnosis has only been reported in the literature a few times. We report on the case of a 42-yr-old female with multiple large metastatic tumor deposits in her liver, pelvic adnexae, bones, and lungs, with an unknown primary. FNA was performed on one of the liver masses, and was diagnosed using routine histochemical and immunohistochemical stains as carcinoid tumor. No follow-up tissue diagnosis has been obtained. However, the patient is still alive with her tumor 1 yr later. Fine-needle aspiration cytology can be a useful and safe tool in the diagnosis of metastatic carcinoid tumors, avoiding the need for surgery which would not otherwise be indicated for treatment.


Infected endovascular graft secondary to coil embolization of endoleak: a demonstration of the importance of operative sterility.
A 60-year-old male underwent endovascular repair of a 5.4-cm enlarging abdominal aortic aneurysm with intraoperative recognition of a type I endoleak. The endoleak was demonstrated to be arising from the left limb of the bifurcated prosthesis. An intravascular stent was placed in the limb near the origin of the common iliac artery and it appeared that the endoleak had sealed. However, 1 month after operation a CT scan demonstrated a persistent, substantial size endoleak without aneurysm enlargement. Coil embolization of the endoleak was undertaken in the interventional radiology suite with apparent satisfactory result. Four days after embolization the patient developed abdominal pain and after 8 days fever and leukocytosis developed. Two weeks after embolization an abdominal CT and indium scan revealed an infected endovascular graft. By CT, the posterior wall of the aneurysm was destroyed and a peri-graft fluid collection with gas was present at the location of the coils. The patient was treated with graft and coil excision and autologous vein reconstruction. Endoluminal prostheses can be contaminated at the time of operative placement. However, an additional source of endoluminal graft infection can arise from secondary endovascular procedures for endoleaks and other graft complications. Since we began placing endovascular grafts at our institution in 1993, most coil embolizations have been performed in the interventional radiology suite. This experience of coil-induced infection causes us to consider performing this type of secondary intervention in the operating room environment.


Oxidation of hydrogen sulfide by human liver mitochondria.
Hydrogen sulfide (H2S) is the third gasotransmitter discovered. Sulfide shares with the two others (NO and CO) the same inhibiting properties towards mitochondrial respiration. However, in contrast with NO or CO, sulfide at concentrations lower than the toxic (μM) level is an hydrogen donor and a substrate for mitochondrial respiration. This is due to the activity of a sulfide quinone reductase found in a large majority of mitochondria. An ongoing study of the metabolic state of liver in obese patients allowed us to evaluate the sulfide oxidation capacity with twelve preparations of human liver mitochondria. The results indicate relatively high rates of sulfide oxidation with a large variability between individuals. These observations made with isolated mitochondria appear in agreement with the main characteristics of sulfide oxidation as established before with the help of cellular models.


A murine model of type 2 diabetes mellitus developed using a combination of high fat diet and multiple low doses of streptozotocin treatment mimics the metabolic characteristics of type 2 diabetes mellitus in humans.
A murine model of type 2 diabetes mellitus was used to compare the antidiabetic effects of the dipeptidyl peptidase-4 (DPP4) inhibitor vildagliptin and biguanide, metformin. Swiss albino mice (n=20 males; n=25 females) were given high fat diet (HFD) ad libitum for 3weeks followed by low dose (40mgkg-1 body weight, bw daily) of streptozotocin (STZ) intraperitoneally five times from the 22nd day of treatment onwards, with HFD continued up to 26th day. Controls (n=15 males; n=15 females) were fed normal balanced diet without administration of STZ. Successful induction of diabetes mellitus was confirmed by testing for fasting blood glucose, intraperitoneal glucose tolerance and intraperitoneal insulin sensitivity. Diabetic mice were administered vildagliptin (10mgkg-1 bw daily) and metformin (50mgkg-1 bw daily) orally for 4weeks. Control, diabetic, vildagliptin and metformin-treated diabetic mice were evaluated for alterations in lipid profile using blood serum and histopathology and oxidative stress using tissues including liver, kidney and heart. Diabetic mice showed significant alterations in lipid profile, tissue histopathology, impaired glucose tolerance, lower insulin sensitivity and elevated lipid peroxidation and protein carbonylation, with depressed catalase activity, when compared to age and gender-matched controls. Metformin and vildagliptin ameliorated the abovementioned diabetic conditions, with vildagliptin found to be more effective. A murine model developed by the combination of HFD and multiple low dose of STZ mimics the metabolic characteristics of type 2 diabetes mellitus in humans, and may be useful for antidiabetic drug screening.


The fate of effete epithelial cells at the villus tips of the human small intestine.
Until recently, little has been known about the morphological features of dying enterocytes at the villus tips of the human small intestine. The present study aimed to show the exfoliating processes of effete enterocytes at the villus tips. Cellular elements of the duodenal lumen and jejunal tissue in humans were fixed and processed for DNA nick end labeling (TUNEL), and transmission and scanning electron microscopy (TEM and SEM). Most cellular elements in the duodenal lumen were enterocytes having TUNEL-positive nuclei. By SEM, protruding enterocytes were discerned at the villus tips. Using the SEM samples embedded in epoxy resin, protruding enterocytes were observed at the villus tips by TEM; they were shrunk by forming numerous clear and autophagic vacuoles, took dome-like profiles, and possessed nuclei with chromatin condensation. The intercellular spaces beneath these protruding or effete enterocytes were often occupied by large lymphocytes. By TUNEL reaction, positive stainings appeared in the epithelium not only at the tip of the villi but also around the site. The results suggest that effete enterocytes at the villus tips of human small intertine are first shrunk by forming clear and autophagic vacuoles, and showed that their nuclei exhibit chromatin condensation immediately before being exfoliated into the lumen.


Sources of lead in Saudi Arabia: a review.
Lead is a poison that affects virtually every system in the body. Children are more vulnerable to lead exposure than adults because of the frequency of pica, hand-to-mouth activity, and a greater rate of intestinal absorption and retention. Very acute lead exposure in children (blood lead levels > 80 microg/dL) can cause coma, convulsions, and even death. Lower levels cause adverse effects on the central nervous system, kidney, and hematopoietic system. Several studies have identified harmful effects of lead in children at blood lead levels as low as 10 microg/dL. Recently, the Center for Disease Control (CDC) has cut the lead allowable threshold level from 25 microg/dL of blood to 10 microg/dL. This review summarizes the main sources of lead exposure that one could expect in a modern society such as Saudi Arabia.


Oral mucosal color changes as a clinical biomarker for cancer detection.
Screening is a key tool for early cancer detection/prevention and potentially saves lives. Oral mucosal vascular aberrations and color changes have been reported in hereditary nonpolyposis colorectal cancer patients, possibly reflecting a subclinical extracellular matrix abnormality implicated in the general process of cancer development. Reasoning that physicochemical changes of a tissue should affect its optical properties, we investigated the diagnostic ability of oral mucosal color to identify patients with several types of cancer. A total of 67 patients with several histologically proven malignancies at different stages were enrolled along with a group of 60 healthy controls of comparable age and sex ratio. Oral mucosal color was measured in selected areas, and then univariate, cluster, and principal component analyses were carried out. Lower red and green and higher blue values were significantly associated with evidence of cancer (all P<0.0001), and efficiently discriminated patients from controls. The blue color coordinate showed significantly higher sensitivity and specificity (96.66±2.77 and 97.16±3.46%, respectively) compared with the red and green coordinates. Likewise, the second principal component coordinate of the red-green clusters discriminated patients from controls with 98.2% sensitivity and 95% specificity (cut-off criterion≤0.4547; P=0.0001). The scatterplots of the chrominances revealed the formation of two well separated clusters, separating cancer patients from controls with a 99.4% probability of correct classification. These findings highlight the ability of oral color to encode clinically relevant biophysical information. In the near future, this low-cost and noninvasive method may become a useful tool for early cancer detection.


One-pot synthesis of amine-substituted aryl sulfides and benzo[b]thiophene derivatives.
A series of amine-substituted aryl sulfides have been synthesized from nitroaryl halides via a simple one-pot procedure involving metal-free C-S cross-coupling and in situ nitro group reduction. Various nitroaryl halides were reacted with thiols in recyclable poly(ethylene glycol) to afford the amine-substituted aryl sulfides in high yield. Additionally, the cross-coupling reactions of nitro- and aldehyde-substituted aryl halides with benzyl thiols under the same reaction conditions were demonstrated to afford benzothiazole and phenylbenzo[b]thiophene derivatives.


Mustard operation in the 1st month of life.
Since April 1976, nine infants less than 1 month of age underwent a Mustard intraatrial baffle procedure. All had transposition of the great arteries and were persistently symptomatic or hypoxic after the Rashkind balloon atrioseptostomy. Two had an associated large ventricular septal defect patched at the time of physiologic correction. The average age of the infants was 11.6 days (range 36 hours to 28 days). Weight averaged 3.4 kg (range 2.5 to 4.2). In all the Mustard procedure was performed using deep hypothermic circulatory arrest, averaging 71 minutes (range 48 to 88) at a mean core temperature of 13 degrees C. An average of 2.8 days of postoperative ventilatory assistance was required. One death occurred in a 28 day old male infant with an associated ventricular septal defect who, the morning after operation, had a sudden unresponsive cardiac standstill. The postoperative hospitalization period averaged 15 days (range 10 to 23). Follow-up evaluation has extended from 4 to 63 months (average 31). Six patients were restudied 2 to 21 months postoperatively. One had obstruction of the superior limb of the baffle, which had separated from the right atrial wall permitting a modest right to left shunt. She has had uneventful repair employing a unique modification of Senning's operation. Another child has asymptomatic partial obstruction of the superior limb of the baffle demonstrated angiographically. None has pulmonary venous obstruction and all have prevailing normal sinus rhythm. Significant right ventricular dysfunction has been demonstrated in one. Early correction in these persistently symptomatic or hypoxic neonates with transposition of the great arteries with or without a ventricular septal defect has obviated the need for palliative operations and produced gratifying early and late results.


[Rhythmic beating cardiomyocytes derived from human embryonic germ (EG) cells in vitro].
Embryonic germ (EG) cells are pluripotent cells derived from primordial germ cells (PGCs) of gonads, gonadal ridges and mesenteries, analogies of fetuses,with the ability to undergo both highly self-renewal and multiple differentiation. These cells in vitro can differentiate into derivatives of all three embryonic germ layers when transferred to an in vitro environment and have the ability to form any fully differentiated cells of the body. The aim of this study is to investigate the potentiality of human EG cells differentiation into cardiomyocytes. Inducing human EG cells with the method of murine ES cells differentiation into cardiomyocytes, supplemented with 0.75%-1% DMSO, 20% NBS, 10(-7) mM RA and 20% cardiomyocytes conditioned medium. 20 heart-like (rhythmic beating cell masses were observed in vitro culture and delayed human EG cells, which beat spontaneously from 20-120 times per minute and maintained beating for 2-15 days, periodic acid's staining (PAS), Myoglobin and a-actin immunological histology positive were all positive and reacted with K+, Ca2+ and adrenalin. Relatively unorganized myofibrillar bundles or more organized sarcomeres, z-bands or a gap junction, the presence of desmosomes in a few cells of the cell masses was observed with transmision electron microscope, which initially demonstrated that these cells were cardiomyocytes. We could not get rhythmly beating cardiomyocytes with 0.75%-1% DMSO, 10-7 mM RA and 20% cardiomyocytes conditioned medium,but in which the percentage of cardiac alpha-actin immunostaining positive cells were increased. The results first demonstrated that human EG cells can differentiate into rhythmic beating cardiomyocytes in vitro and suggests that human EG cells may represent a new potent resource for cardiomyocytes transplantation therapy for myocardium infarction.


Dual signaling functions of the hybrid sensor kinase RpfC of Xanthomonas campestris involve either phosphorelay or receiver domain-protein interaction.
The hybrid sensor kinase RpfC positively regulates the expression of a range of virulent genes and negatively modulates the synthesis of the quorum sensing signal diffusible signal factor (DSF) in Xanthomonas campestris. Three conserved amino acid residues of RpfC implicated in phosphorelay (His(198) in the histidine kinase domain, Asp(512) in the receiver domain, and His(657) in the histidine phosphotransfer domain) were essential for activation of the production of extracellular enzymes and extracellular polysaccharide (EPS) virulence factors but were not essential for repression of DSF biosynthesis. Domain deletion and subsequent in trans expression analysis revealed that the receiver domain of RpfC alone was sufficient to repress DSF overproduction in an rpfC deletion mutant. Further deletion and alanine scanning mutagenesis analyses identified a peptide of 107 amino acids and three amino acid residues (Gln(496), Glu(504), and Ile(552)) involved in modulating DSF production. Co-immunoprecipitation and far Western blot analyses suggested an interaction between the receiver domain and RpfF, the enzyme involved in DSF biosynthesis. These data support a model in which RpfC modulates two different functions (virulence factor synthesis and DSF synthesis) by utilization of a conserved phosphorelay system and a novel domain-specific protein-protein interaction mechanism, respectively. This latter mechanism represents an added dimension to conventional two-component signaling paradigms.


Pharmacological evidence for high affinity and low affinity a2-adrenoceptor binding sites in rat vas deferens.
Differential antagonistic activity against imidazoline- and phenethylamine-induced inhibition of field stimulated rat vas deferens is described. Competitive antagonism of the imidazolines, chlonidine and B-HT 920, was produced by the a2-selective antagonists yohimbine and phentolamine, whereas, only partial antagonism of a-methyl noradrenaline and adrenaline was observed. Higher concentrations of yohimbine (above 2 microM) and phentolamine (above 20 microM) failed to produce a further shift in the dose-response curves of these agonists. The rate of recovery of the twitch response following clonidine- or B-HT 920-induced inhibition was very slow, even after repeated washing of the tissue. On the other hand recovery following a-methyl noradrenaline or adrenaline was extremely rapid. The present results provide pharmacological evidence in support of the previously proposed existence of high and low affinity binding site on the a2-adrenoceptor.


Single-Cell Transcriptomics of Traced Epidermal and Hair Follicle Stem Cells Reveals Rapid Adaptations during Wound Healing.
Epithelial tissues, such as the skin, rely on cellular plasticity of stem cells (SCs) from different niches to restore tissue function after injury. How these molecularly and functionally diverse SC populations respond to injury remains elusive. Here, we genetically labeled Lgr5- or Lgr6-expressing cells from the hair follicle bulge and interfollicular epidermis (IFE), respectively, and monitored their individual transcriptional adaptations during wound healing using single-cell transcriptomics. Both Lgr5 and Lgr6 progeny rapidly induced a genetic wound signature that, for Lgr5 progeny, included the remodeling of receptors to permit interactions with the wound environment, a property that Lgr6 progeny possessed even before wounding. When contributing to re-epithelialization, Lgr5 progeny gradually replaced their bulge identity with an IFE identity, and this process started already before Lgr5 progeny left the bulge. Altogether, this study reveals how different SCs respond and adapt to a new environment, potentially explaining cellular plasticity of many epithelial tissues.


[Toxic shock like syndrome due to S. pyogenes (TSLS)].
Resurgence of severe infection, caused by Streptococcus pyogenes was observed in the USA from the middle of the 1980s. Toxic Shock like Syndrome due to Streptococcus pyogenes (TSLS) has been one of the most life-threatening diseases with soft tissue infections, such as necrotizing fasciitis, myositis, bacteremia and shock. Patients with TSLS has been reported in Europe, USA, Canada, etc. The first report of TSLS in Japan was described by Y. Shimizu and his colleague, Asahi General Hospital, Chiba, 1992. Since then more cases of TSLS, diagnosed by CDC's Criteria, have been found in various parts of Japan, including Chiba prefecture. They are characterised by rapid development of shock and death. Some of the isolated strains from blood or tissues have been identified as S. pyogenes, M3, M1 with SPEA (Streptococcal pyrogenic exotoxin A).


Preferential increase in chylomicron levels of the xanthophylls lutein and zeaxanthin compared to beta-carotene in the human.
The time-course of appearance of five carotenoids in chylomicrons and their distribution pattern were studied following ingestion of a single dose of Betatene, a natural carotenoid source composed of 0.5% lutein, 0.75% zeaxanthin, 3.6% alpha-carotene, 70.3% all-trans beta-carotene, 22.7% beta-carotene cis isomers, 2.1% unidentified carotenoids and no lycopene. Lutein, zeaxanthin, alpha-carotene and all-trans beta-carotene but not lycopene levels in chylomicrons increased after Betatene ingestion with a maximum at 9 h and a distinct decline from 9 to 12 h. However, the carotenoid pattern in the chylomicron fraction did not match the pattern in Betatene, with a 14-fold and 4-fold higher level of lutein and zeaxanthin, respectively, compared to their content in Betatene. The contribution of alpha-carotene to the carotenoid composition in chylomicrons reflected its content in Betatene, whereas the relative amount of all-trans beta-carotene was substantially lower than in Betatene. Thus, in the presence of high amounts of beta-carotene, there is a preferential uptake of the xanthophylls lutein and zeaxanthin as compared to all-trans beta-carotene from the intestinal lumen into chylomicrons.


Effect of voluntary wheel-running on insulin sensitivity and responsiveness in high-fat-fed rats.
The effect of voluntary wheel-running on insulin resistance was studied in high-fat-fed rats. A sequential hyperinsulinemic euglycemic clamp procedure was employed (insulin infusion rates: 3 and 30 mU/kg BW/min) in 14 high-fat-fed rats and 7 chow-fed rats under the awake condition. The high-fat-fed rats were further divided into a sedentary (n=7) and a voluntary wheel-running (n=7) groups. Blood glucose was clamped at the fasting level in each rat. Plasma insulin levels during the 3- and 30-mU/kg BW/min insulin infusions were 40-50 and 450-550 microU/ml, respectively. At both 3 and 30 mU/kg BW/min insulin infusions, high-fat-feeding showed a significant decrease in glucose infusion rate (GIR), compared with the chow-fed rats. However, decreased GIRs were restored by the 4-wk wheel-running and reached similar levels as the chow-fed rats. Therefore, it could be concluded that voluntary wheel-running prevents insulin resistance induced by high-fat feeding.


An advanced cone-and-plate reactor for the in vitro-application of shear stress on adherent cells.
Endothelial cells (ECs) are permanently exposed to the blood flow and the resulting shear stress, its magnitude varying with the EC site in the blood stream. Along with other mechanical stimuli like vessel wall stretching or hydrostatic blood pressure, this shear stress modulates the endothelial cell function, morphology and gene expression. Here, we describe our improved cone-and-plate reactor that applies up to 10 dyn/cm(2) uniform wall shear stress on a defined, ring-shaped region on a culture dish. At the same time, a hydrostatic pressure of up to 195 mmHg can be applied by increasing the atmospheric pressure in the incubator box. Gas composition can be controlled additionally, used for maintaining CO2-homeostasis or inducing hypoxic conditions. For better comparability, six cone-and-plate systems can be used at the same time at different rotational velocities. The effects on cell morphology, cytoskeleton and cell alignment can be monitored during application using a laser scanning microscope. Flow conditions have been studied and a sufficient area of uniform wall shear stress could be shown. To exceed 10 dyn/cm2, we suggest an increase in medium viscosity.


Induction of competence to respond to IL-4 by CD4+ T helper type 1 cells requires costimulation.
Rested murine CD4+ Th1 clones do not produce IL-4, but have previously been shown to be capable of responding to IL-4 if they are first activated with Ag and APC. In this study, we have examined the activation requirements for induction of competence to respond to IL-4 in these clones. TCR occupancy alone (given either as chemically fixed APC and Ag, anti-CD3, Con A, or ionomycin and PMA) was inadequate, but the addition of a source of costimulation to any of these stimuli resulted in complete induction of competence to respond to IL-4. Pretreatment of the Th1 clones with TCR occupancy alone induced an anergic state from which subsequent full stimulation with Ag and APC failed to give IL-4 responsiveness. Pretreatment of the cells with IL-2 alone was an inadequate signal to induce IL-4 responsiveness and only a partial response was obtained when TCR occupancy was combined with IL-2. Addition of anti-IL-2 and anti-IL-2R antibodies during full activation with APC and Ag gave a 50% inhibition of competence induction. These results demonstrate that costimulation, in addition to its role in IL-2 production, is an important second signal for inducing T cells to become competent to respond to IL-4.


Back pain in psoriatic arthritis: defining prevalence, characteristics and performance of inflammatory back pain criteria in psoriatic arthritis.
We aimed to determine the agreement between rheumatologist-judged inflammatory back pain (IBP) and criteria defining IBP in patients with psoriatic arthritis (PsA) and predictive value of IBP in identifying axial involvement in PsA. Using prospectively collected data, we investigated the agreement between rheumatologist judgement of IBP and IBP criteria (Calin, Rudwaleit and Assessment of Spondyloarthritis International Society) using the kappa coefficient. We also determined the sensitivity, specificity and likelihood ratios of the presence of back pain, rheumatologist-judged IBP and the three IBP criteria for detecting axial PsA (AxPsA). Finally, we compared the clinical and genetic markers in patients with PsA with axial radiological changes with and without back pain. 171 patients (52% male, mean age 46.6 years) were identified. Ninety-six (56.13%) patients reported chronic back pain. Sixty-five (38.01%) had IBP. 54 (32%) patients had evidence of radiological change in the spine. The agreement between rheumatologist judgement of IBP and IBP criteria was highest for the Calin criteria (0.70). Positive likelihood ratio for the presence of radiological axial involvement was highest for Rudwaleit criteria (2.17). No differences between patients with AxPsA with or without back pain were found, except for higher Bath Ankylosing Spondylitis Disease Activity Index and lower prevalence of human leucocyte antigen-B*38 in those with back pain. Rheumatologist-judged IBP or the criteria for IBP developed for ankylosing spondylitis may not perform well when ascertaining axial involvement in PsA.


Retinal polarization effects.
Retinal photographs taken with crossed polarizers in the input and recording light paths show a cross or brush pattern overlying the macula. Experiments were done indicating that this pattern is due to the birefringence in cone-photoreceptor outer segments. Haidinger's brushes are also attributed to cone outer segments, but the effect is due to a combination of the dispersion in the birefringence and the dichroism of the outer segments. The objective, polarized-light retinal pattern is shown to be a useful clinical tool for diagnosing diseases affecting the macula.


Oxytocin receptor gene expression of estrogen-stimulated human myometrium in extracorporeally perfused non-pregnant uteri.
Oxytocin (OT) and the oxytocin receptor (OTR) seem to be less important for uterine contractility-associated disorders of the non-pregnant uterus compared to the pregnant uterus. In the present study, we investigated the mutual dependence of OTR, OT and 17beta-estradiol (E(2)) with regard to the localization of OTR in the non-pregnant uterus. Utilizing our established model for extracorporeal perfusion of the human uterus, we perfused 15 human uteri for 27 h under physiological conditions without oestradiol (group A, n = 5) or with high E(2) stimulation (group B, n = 5) followed by OT stimulation for both groups during the last 3 h of the experiment. Negative controls (n = 5) remained in perfusions for 27 h without any further hormone treatment. Gene expression of the myometrial OTR in both groups was compared using reverse transcriptase triple primer PCR. Stimulation with E(2) and OT led to significantly higher OTR gene expression than stimulation with OT alone. We also showed that concentrations of OTR transcripts increase from the lower uterine segment to the uterine fundus. However, maximum OTR levels of the uterine fundus in group B did not reach concentrations of specimens of third trimester of pregnancy which were used as positive controls. We conclude that our experimental model simulates a situation similiarly to the stimulated non-pregnant uterus in the therapeutic concepts of assisted reproduction. The data presented demonstrate that the dynamics of OTR expression can be modulated by stimulation with E(2) and OT, not only in the pregnant but also in the non-pregnant uterus.


Rapamycin protects the mitochondria against oxidative stress and apoptosis in a rat model of Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disease, in which oxidative stress and mitochondrial dysfunction are responsible for neuronal apoptosis. Rapamycin plays a crucial role in reducing oxidative stress and protecting the mitochondria. However, its protective role in PD has not yet been fully elucidated. In this study, we report that pre-treatment with rapamycin provides behavioral improvements, protects against the loss of dopaminergic neurons, and alleviates mitochondrial ultrastructural injuries in a rat model of PD. Peroxide levels were lower and antioxidant activities were higher in PD rats pre-treated with rapamycin compared to the PD rats pre-treated with the vehicle. Furthermore, pre-treatment with rapamycin significantly elevated the expression of anti-apoptotic markers and reduced the levels of pro-apoptotic markers compared to pre-treatment with the vehicle. In conclusion, our results demonstrated that rapamycin reduced oxidative stress and alleviated mitochondrial injuries in the 6-hydroxydopamine (6-OHDA)-induced rat model of PD, which may subsequently contribute to its anti-apoptotic effects. The ability of rapamycin to exhibit neuroprotection in a rat model of PD may be related to its antioxidant capabilities.


Micropatterned surfaces prepared using a liquid crystal projector-modified photopolymerization device and microfluidics.
A commercial liquid crystal device projector was modified for photopolymerization using its on-board intense light source and a precision optical control circuit. This device projects reduced images generated by a typical personal computer onto the stage where photopolymerization on a surface occurs. This all-in-one device does not require expensive photomasks and external light sources. However, light scattering and diffraction through glass substrates resulted in undesired reactions in areas corresponding to masked (black) domains in mask patterns, limiting pattern resolution. To overcome this shortcoming, two-step surface patterning was developed. First, three-dimensional microstructures of crosslinked silicone elastomer were fabricated with this device and adhered onto silanized glass substrate surfaces, forming microchannels in patterns on the glass support. Then, acrylamide monomer solution containing photoreactive initiator was flowed into these micromold channels and reacted in situ. The resultant polyacrylamide layer was highly hydrophilic and repelled protein adsorption. Cell seeding on these patterns in serum-supplemented culture medium produced cells selectively adhered to different patterns: cells attached and spread only on unpolymerized silanized glass surfaces, not on the photopolymerized acrylamide surfaces. This technique should prove useful for inexpensive, rapid prototyping of surface micropatterns from polymer materials.


Hand osteoarthritis in Chuvashian population: prevalence and determinants.
The objectives of this study are to evaluate the prevalence of radiographic hand osteoarthritis (OA) in Chuvashian community-based sample and its associations with age, sex, body mass index (BMI), smoking, and alcohol consumption. The study population was comprised of Chuvashians (443 males, age 47.4 ± 17.0 years and 384 females, age 50.8 ± 16.3 years). OA was evaluated for 14 joints of each hand according to the Kellgren and Lawrence grading scheme. Statistical analyses included prevalence estimation, linear, logistic and polynomial regressions, and χ2 tests. After the age of 65 years, 89.2% of males and 97.6% of females had at least one affected joint. For males and females the best-fitted and most parsimonious model of association between age and number of affected hand joints was the two interval linear one. After adjustment for age, no association between prevalence of hand OA and BMI was found in males. However, females with obesity have almost three times higher risk of development of hand OA than those with normal weight. No association was found between smoking and prevalence of hand OA. Moderate alcohol consumption showed protective effect on prevalence of hand OA in women.


Responses of seasonally anovulatory mares to daily administration of thyrotropin-releasing hormone and(or) gonadotropin-releasing hormone analog.
Seventeen seasonally anovulatory light horse mares were treated daily, starting January 5 (d 1), for 28 d with GnRH analog (GnRH-A; 50 ng/kg BW) and(or) thyrotropin-releasing hormone (TRH; 5 microg/kg BW) in a 2 x 2 factorial arrangement of treatments to test the hypothesis that combined treatment may stimulate follicular growth and development. Ovaries were examined via ultrasonography and jugular blood samples were collected every 3 d. Frequent blood samples were collected after treatment injections on d 1, 2, 4, 7, 11, 16, and 22; on d 29, all mares received an i.v. mixture of GnRH, TRH, sulpiride, and EP51389 (a growth hormone secretagogue) to assess pituitary responsiveness. No consistent effects (P > 0.1) of treatment were observed for plasma LH, FSH, prolactin, or thyroxine concentrations in samples collected every 3 d. The only effect on ovarian follicle numbers was a reduction in number of follicles 11 to 19 mm in diameter due to TRH treatment (P = 0.029). No mare ovulated during treatment. On the days of frequent sampling, mean LH (P = 0.0001) and FSH (P = 0.001) concentrations were higher in mares receiving GnRH-A and tended to increase from d 1 through 7. In contrast, mean prolactin (P = 0.001) and thyroid-stimulating hormone (P = 0.0001) concentrations were high in mares receiving TRH on d 1 but rapidly decreased thereafter. When mares were administered the secretagogue mixture on d 29, the LH response was greater (P = 0.0002) in mares that had previously received GnRH-A but the FSH response was not affected (P > 0.1); the prolactin response was greater (P = 0.014) and the TSH response was smaller (P = 0.0005) in mares that had previously received TRH. Surprisingly, an immediate growth hormone response to EP51389 was absent in all mares. In conclusion, daily GnRH-A treatment stimulated plasma LH and FSH concentrations immediately after injection; although no long-term elevation in preinjection concentrations was achieved, the responses gradually increased over time, indicating a stimulation of gonadotropin production and storage. Daily treatment with TRH stimulated plasma TSH and prolactin concentrations, but the response diminished rapidly and was minimal within a few days, indicating a depletion of pituitary stores and little or no stimulation of production. There was no beneficial effect of adding TRH treatment to the daily GnRH-A regimen.


Influence of Liquid Structure on Fickian Diffusion in Binary Mixtures of n-Hexane and Carbon Dioxide Probed by Dynamic Light Scattering, Raman Spectroscopy, and Molecular Dynamics Simulations.
This study contributes to a fundamental understanding of how the liquid structure in a model system consisting of weakly associative n-hexane ( n-C6H14) and carbon dioxide (CO2) influences the Fickian diffusion process. For this, the benefits of light scattering experiments and molecular dynamics (MD) simulations at macroscopic thermodynamic equilibrium were combined synergistically. Our reference Fickian diffusivities measured by dynamic light scattering (DLS) revealed an unusual trend with increasing CO2 mole fractions up to about 70 mol %, which agrees with our simulation results. The molecular impacts on the Fickian diffusion were analyzed by MD simulations, where kinetic contributions related to the Maxwell-Stefan (MS) diffusivity and structural contributions quantified by the thermodynamic factor were studied separately. Both the MS diffusivity and the thermodynamic factor indicate the deceleration of Fickian diffusion compared to an ideal mixture behavior. Computed radial distribution functions as well as a significant blue-shift of the CH stretching modes of n-C6H14 identified by Raman spectroscopy show that the slowing down of the diffusion is caused by a structural organization in the binary mixtures over a broad concentration range in the form of self-associated n-C6H14 and CO2 domains. These networks start to form close to the infinite dilution limits and seem to have their largest extent at a solute-solvent transition point at about 70 mol % CO2. The current results not only improve the general understanding of mass diffusion in liquids but also serve to develop sound prediction models for Fick diffusivities.


Acute changes in cellular zinc alters zinc uptake rates prior to zinc transporter gene expression in Jurkat cells.
A coordinated network of zinc transporters and binding proteins tightly regulate cellular zinc levels. Canonical responses to zinc availability are thought to be mediated by changes in gene expression of key zinc transporters. We investigated the temporal relationships of actual zinc uptake with patterns of gene expression in membrane-bound zinc transporters in the human immortalized T lymphocyte Jurkat cell line. Cellular zinc levels were elevated or reduced with exogenous zinc sulfate or N,N,N',N-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), respectively. Excess zinc resulted in a rapid 44 % decrease in the rate of zinc uptake within 10 min. After 120 min, the expression of metallothionein (positive control) increased, as well as the zinc exporter, ZnT1; however, the expression of zinc importers did not change during this time period. Zinc chelation with TPEN resulted in a rapid twofold increase in the rate of zinc uptake within 10 min. After 120 min, the expression of ZnT1 decreased, while again the expression of zinc importers did not change. Overall, zinc transporter gene expression kinetics did not match actual changes in cellular zinc uptake with exogenous zinc or TPEN treatments. This suggests zinc transporter regulation may be the initial response to changes in zinc within Jurkat cells.


Fractionation of a hydrocolloid emulsifier reclaimed from winery waste.
Complex hydrocolloids have been isolated and fractionated using a consecutive elution process, starting from winery waste. These extracts consist mainly of polysaccharidic populations and of smaller protein molecules and they exhibit emulsifying, thickening and texture-modifying activity. This work is a systematic study of these individual populations, as fractionated with preparative size exclusion chromatography (Prep-SEC) in terms of their chemical identity, surface properties, and emulsification behavior. The fractions have been characterized via SEC-MALLS, FTIR, DLS, zeta potential, and interfacial tension measurements. The results highlight the antagonistic and synergistic effects of the individual components of the above-mentioned complex natural material (winery waste extract) towards its emulsifying behavior, and provide a model for the kinetics of the evolution of a Pickering interfacial layer.


Variables affecting outcome in blunt chest trauma: flail chest vs. pulmonary contusion.
We reviewed 144 consecutive patients with flail chest and/or pulmonary contusion between 1979 and 1984. The purpose was to analyze the factors adversely affecting morbidity and mortality. There were 97 males and 47 females, with an average age of 40 years +/- 18 S.D. (range, 2-83). Seventy-five per cent of the injuries were caused by motor vehicle accidents, with the remainder due to falls (17%), cardiopulmonary resuscitation (4%), altercations (2%), or falling objects (2%). The Injury Severity Score (ISS) averaged 32 +/- 14 S.D. in all survivors versus 60 +/- 14 S.D. in those who died. Eighty-three patients (58%) required mechanical ventilation. Thirty-six patients died (25%). Isolated pulmonary contusion or flail chest had a mortality of 16% each. However, the mortality more than doubled when there was a combined pulmonary contusion and flail chest (42%). More than half of all deaths were directly attributed to central nervous system injuries with another third due to massive hemorrhage. Factors that were associated with a higher morbidity and mortality included severe associated thoracic injuries, a high ISS, the presence of shock, falls from heights, and the combination of pulmonary contusion and flail chest.


Warming Accelerates Carbohydrate Consumption in the Diapausing Overwintering Peach Fruit Moth Carposina sasakii (Lepidoptera: Carposinidae).
Climate warming provides a challenge for small insects persisting in cold seasons through diapause because they fail to accumulate and maintain adequate reserves to complete this stage successfully. One way of understanding this challenge is to follow physiological changes in these insects under higher temperatures, including the consumption and allocation of energy reserves during and after diapause. We simulated autumn and spring warming conditions to study carbohydrate consumption dynamics during diapause-post-diapause periods by monitoring shifts in carbohydrate levels in a facultative diapause species, the peach fruit moth Carposina sasakii Matsumura (Lepidoptera: Carposinidae). We found carbohydrates were rapidly consumed in the post-diapause phase, which might lead to a trade-off in the allocation of energy reserves between diapause maintenance and post-diapause development. This suggests that temperature increases in autumn and spring may alter diapause maintenance and post-diapause development through changing carbohydrate levels.


Delayed cone-opponent signals in the luminance pathway.
Cone signals in the luminance or achromatic pathway were investigated by measuring how the perceptual timing of M- or L-cone-detected flicker depended on temporal frequency and chromatic adaptation. Relative timings were measured, as a function of temporal frequency, by superimposing M- or L-cone-isolating flicker on "equichromatic" flicker (flicker of the same wavelength as the background) and asking observers to vary contrast and phase to cancel the perception of flicker. Measurements were made in four observers on up to 35 different backgrounds varying in wavelength and radiance. Observers showed substantial perceptual delays or advances of L- and M-cone flicker that varied systematically with cone class, background wavelength, and radiance. Delays were largest for M-cone-isolating flicker. Although complex, the results can be characterised by a surprisingly simple model in which the representations of L- and M-cone flicker are comprised not only of a fast copy of the flicker signal, but also of a slow copy that is delayed by roughly 30 ms and varies in strength and sign with both background wavelength and radiance. The delays, which are too large to be due to selective cone adaptation by the chromatic backgrounds, must arise postreceptorally. Clear evidence for the slow signals can also be found in physiological measurements of horizontal and magnocellular ganglion cells, thus placing the origin of the slow signals in the retina-most likely in an extended horizontal cell network. Luminance-equated stimuli chosen to isolate chromatic channels may inadvertently generate slow signals in the luminance channel.


[Quantitation of regional cerebral blood flow using 123I-IMP from a single SPECT scan and a single blood sampling--analysis on statistical error source and optimal scan time].
Recently, a method has been proposed to measure quantitative regional cerebral blood flow (rCBF) in man using N-isopropyl-p-[123I]-iodoamphetamine (IMP) and single photon emission tomography (SPECT). In this method (IMP-ARG method), a functional map of rCBF was calculated from a single SPECT scan data set, in which effects of the clearance of IMP from the brain was taken into account by employing the 2 compartment model. To avoid the procedures of frequent and/or continuous sampling of the arterial blood that was required in determination of the input function, use of a standard input function and calibrating it by one blood sampling were validated. The present study was intended to investigate the sensitivity to various sources of errors in the IMP-ARG method such as (1) effects of individual difference of the arterial input function, (2) effects of ambiguity of the regional distribution volume (Vd) of IMP, and (3) effects of inaccuracy of the SPECT measurement. It was shown in the present simulation study that errors in the calculated rCBF values were dependent on the SPECT mid-scan time (MST), and that the minimum error corresponded to the MST of approximately 30 min after the IMP infusion. With this optimal MST, errors in rCBF due to the individual difference of the input function was approximately 10%, and the errors due to the ambiguity of the Vd were approximately 8%. The total accuracy of the calculated rCBF in the IMP-ARG method was, therefore, estimated as approximately 13%. Although a relatively large error was expected at a high rCBF area, this study strongly suggested the IMP-ARG method being useful and accurate for providing the quantitative rCBF map for a clinical use.


Macrophages-induced long noncoding RNA H19 up-regulation triggers and activates the miR-193b/MAPK1 axis and promotes cell aggressiveness in hepatocellular carcinoma.
Dysregulation of long noncoding RNA (lncRNA) H19 has been implicated in hepatocellular carcinoma (HCC), but the concrete regulatory mechanism is lack of research. We mined gene expression profiles of 457 HCC samples from TCGA and TJMUCH cohorts and further validated in 64 FFPE HCC tissues. LncRNA H19 overexpression in situ was significantly correlated with poor prognosis of HCC patients, which induced EMT, promoted stemness and accelerated invasion of HCC cells in vitro. Co-expression network analysis indicated lncRNA H19 negatively correlated with miR-193b and positively correlated with MAPK1 gene, which implicated that lncRNA H19 served as a sponge molecule to hijack miR-193b and protect MAPK1. Forced overexpression of H19 attenuated miR-193b-mediated inhibition on multiple driver oncogenes (EGFR, KRAS, PTEN and IGF1R) and MAPK1 gene, thus triggered EMT and stem cell transformation in HCC. LncRNA H19 positively correlated with CD68 + TAMs in situ. TAMs-induced lncRNA H19 promotes HCC aggressiveness via triggering and activating the miR-193b/MAPK1 axis, mediates the crosstalk between HCC and immunological microenvironment, and causes poor clinical outcomes. LncRNA H19 is a valuable predictive biomarker and potential therapeutic target in HCC.


Prophylaxis against opportunistic infections in persons infected with human immunodeficiency virus.
This article outlines the current official recommendations for the prevention of opportunistic disease in adults and adolescents infected with human immunodeficiency virus, including specific guidelines for discontinuing primary and secondary prophylaxis when immune reconstitution has occurred as a result of highly active antiretroviral therapies. The recommendations, developed by the U. S. Public Health Service and the Infectious Diseases Society of America for clinicians and healthcare providers, were originally published in 1995 and revised in 1997, 1999, and 2002. The 2002 recommendations are summarized in this article.


Rapid alterations of BDNF protein levels in the rat brain after focal ischemia: evidence for increased synthesis and anterograde axonal transport.
Cellular localization and tissue levels of BDNF protein were studied using immunocytochemistry and enzyme immunoassay, respectively, in the cortex and striatum at different reperfusion times (0-24 h) after 2 h of unilateral middle cerebral artery occlusion (MCAO) in rats. The distribution of neuronal injury was analyzed in NeuN-, cresyl violet-, and Fluoro-Jade-stained sections. At 2 h postischemia, but not at later time points, there was a several-fold increase of the number of BDNF-immunoreactive (-ir) cells in the ipsilateral cingulate and frontal cortices outside the damaged area. Animals with cortical injury showed loss of BDNF-ir fibers in the striatum at 2-24 h, whereas rats with cell damage confined to the striatum exhibited no such change. At 2-16 h, strongly BDNF-ir fibers were observed along the myelinated fascicles medially in the striatum, in the anterior commissure, and in the corpus callosum ipsilateral to the MCAO. BDNF protein levels were increased (by 133-213%) at 2 h in the cingulate and frontal cortices and decreased (by 40%) at 24 h in the striatum. These findings show that the increased expression of BDNF mRNA in cortical neurons previously demonstrated after transient focal ischemia is accompanied by elevated levels of BDNF protein. The rapid decline of BDNF protein levels suggests a pronounced release or anterograde axonal transport in the postischemic phase. The reduction of BDNF protein in the striatum of animals with cortical damage provides further evidence for anterograde transport, which is also supported by the accumulation of BDNF protein in several preterminal fiber systems. The changes of BDNF protein after focal ischemia could play a role for survival and plasticity of cortical and striatal neurons.


When Frail Individuals or Their Families Request Nonindicated Interventions: Usefulness of the Four-Box Ethical Approach.
In the age of person-centered care, there is an emphasis on promoting patient autonomy and surrogate decision maker authority in making treatment decisions that are aligned with the patient's priorities and values. As technological advances offer multiple clinical options with various levels and types of risks and benefits, person-centered clinical practice encourages the incorporation of patients' and families' heterogeneous experiences into decisions regarding illness management. In caring for frail elderly adults, clinicians are sometimes faced with situations in which individuals and their surrogate decision-makers request a treatment that the clinicians feel is clinically inappropriate. This article provides a case example of a frail older adult with advanced chronic kidney disease who requests dialysis despite the advice of his nephrologist to pursue conservative management. The four-box approach, which provides clinicians with a structured ethical framework to facilitate informed and ethically justified treatment decisions, is then introduced. By considering the patient's medical indications, preferences, quality of life, and contextual factors, how each consideration plays a unique yet equally important role in informing clinically responsible and person-centered care is illustrated.


Cyanide--an uncompetitive inhibitor of NAD(P)-dependent malic enzyme from human term placental mitochondria.
The activity of NAD(P)-dependent mitochondrial malic enzyme was considerably inhibited by KCN, whereas under the same conditions azide affected the enzyme only slightly. Kinetic studies showed that KCN is an uncompetitive inhibitor of mitochondrial malic enzyme from human placenta. In contrast to the mitochondrial enzyme, the cytosolic malic enzyme was only slightly affected by KCN and under the same conditions the effect of azide was negligible. The effect of KCN was compared to this on the malic enzyme from other sources.


Effect of adenosine and adenosine 5'-monophosphate on cell division of cultured mastocytoma P-815 cells.
The growth of mouse mastocytoma P-815 cells in culture (37 degrees, 42 hr) was inhibited by exogenous adenosine (0.2 to 1.0 mM) and more effectively by AMP (0.01 to 0.1 mM), but not by adenine. The inhibited growth (a 25% inhibition by 0.5 mM adenosine and a 80% inhibition by 0.25 mM AMP) was restored to a near control level by the addition of uridine (0.5 mM) to the medium. The pretreatment (37 degrees, 3 hr) of the cells with adenosine or AMP caused a 60% inhibition of incorporation (37 degrees, 2 hr) of [U-14C]aspartate into uracil nucleotides, accumulating 14C-orotate and orotidine. Both dipyridamole, an inhibitor of adenosine uptake, and exogenous adenosine deaminase suppressed the growth inhibition induced by not only adenosine but also AMP. 2-Chloroadenosine, which is resistant to the action of adenosine deaminase, was a more potent growth inhibitor, while 3'AMP and 2'-AMP, which are not hydrolyzed to adenosine by membrane 5'-nucleotidase, were ineffective. Adenosine 5'-sulfate and other 5'-substituted adenosines were also ineffective. These observations indicate that AMP inhibits the growth of mastocytoma P-815 cells as a result of its continuous conversion to adenosine and a constant exposure of the cells to a low concentration of adenosine which readily permeates the cell membrane. In addition, adenosine, AMP and their agarose-linked forms rapidly (37 degrees, 20 min) elevated cellular levels of cAMP. This effect was not suppressed by dipyridamole. Apparently adenosine and AMP also act extracellularly for growth inhibition by regulating cAMP levels.


Extracellular chaperones.
The maintenance of the levels and correct folding state of proteins (proteostasis) is a fundamental prerequisite for life. Life has evolved complex mechanisms to maintain proteostasis and many of these that operate inside cells are now well understood. The same cannot yet be said of corresponding processes in extracellular fluids of the human body, where inappropriate protein aggregation is known to underpin many serious diseases such as Alzheimer's disease, type II diabetes and prion diseases. Recent research has uncovered a growing family of abundant extracellular chaperones in body fluids which appear to selectively bind to exposed regions of hydrophobicity on misfolded proteins to inhibit their toxicity and prevent them from aggregating to form insoluble deposits. These extracellular chaperones are also implicated in clearing the soluble, stabilized misfolded proteins from body fluids via receptor-mediated endocytosis for subsequent lysosomal degradation. Recent work also raises the possibility that extracellular chaperones may play roles in modulating the immune response. Future work will better define the in vivo functions of extracellular chaperones in proteostasis and immunology and pave the way for the development of new treatments for serious diseases.


The effects of an electronic order set on vancomycin dosing in the ED.
The objective of the study was to assess the impact of a computer physician order entry (CPOE) electronic order set on appropriate vancomycin dosing in the emergency department (ED). We conducted a retrospective study examining ED dosing of vancomycin before and after the implementation of an electronic weight-based vancomycin order set. Preimplementation and postimplementation patient records were analyzed between the dates of June 1st and August 31st 2010 for the pre-CPOE group and January 1st to March 31st 2013 for the post-CPOE group. χ(2) analysis, Fisher exact test, and t tests were performed with a 2-sided P value <.05 denoting statistical significance, where appropriate. A total of 597 patients were included in the study, with 220 in the pre-CPOE group and 377 in the post-CPOE group. The use of the electronic order set resulted in a 21.9% increase (P < .05) in appropriate dosing with 67.4% (254/377) of post-CPOE vancomycin doses considered appropriate vs 45.5% (100/220) in the pre-CPOE group. In critically ill patients, there was a 16.3% increase in appropriate dosing with 44.7% (38/85) in the post-CPOE group compared with 28.4% (19/67) in the pre-CPOE group. The implementation of an electronic order set increased the percentage of ED patients receiving appropriate initial vancomycin doses. The impact of increasing compliance to vancomycin guidelines is in accordance with stewardship principles that promote optimization of antimicrobial dosing based on individual patient characteristics. More studies are needed to assess the relationship between appropriate vancomycin loading doses in the ED and therapeutic outcomes.


Evaluation of the effectiveness of a phonoaudiology program for the rehabilitation of dysphagia in the elderly.
The object of this study was to evaluate the effectiveness of a phonoaudiology rehabilitation program directed towards the degenerative changes of the neurophysiological mechanisms responsible for the upper digestive system compromised by the age between 80 and 90 years, in a regime of internment in a long permanence institution in the city of Rio de Janeiro, which express, bodily, their difficulty in swallowing food of liquid or pasty consistency. To achieve such an object, the current study was developed in an experimental format or design, comprised of a sample group of 23 elderly subjects, of both genders, undergoing evaluation by the phonoaudiology protocol for evaluating the risk of dysphagia (abbreviated from the Portuguese name: Protocolo de Avaliação do Risco para Disfagia=PARD), pre- and post-intervention procedure. The developed program used the indirect therapy approach based on the adaptive myotherapeutic and myofunctional stimulation, contemplating two isometric and isokinetic orofacial active myotherapeutic exercises, two passive myotherapeutic manipulations for the supra-hyoidal musculature, as well as adaptive cephalic postural maneuvres, associated to the intra-oral olfactive and gustative sensory stimulation. The evolution of the individuals was analyzed before and after the therapy by estimating the capacity of swallowing food in the liquid fluid (LF) consistency, by the paired Student t-test, giving a p=0.01, making the difference of 2.31 between the 2 evaluations statistically significant. The changes produced by the PARD program in the capacity of individuals in swallowing food of a pasty consistency were also significant when analyzing the variance of the results on the 2 instances by using the F-test (pre- and post-intervention difference=4.47, p=0.039).


Posttraumatic shame and guilt.
Posttraumatic shame and guilt are important dimensions of posttraumatic syndromes, including simple and complex post-traumatic stress disorder (PTSD). The concepts of posttraumatic shame and guilt have received little theoretical or empirical investigation in the field of traumatology. It is proposed that there are acute and prolonged states of posttraumatic shame and guilt that can be compared in their consequences across eight psychosocial dimensions: (a) self-attribution processes, (b) emotional states and capacity for affect regulation, (c) appraisal and interpretation of actions, (d) the impact of states of shame and guilt on personal identity, (e) suicidality, (f) defensive patterns, (g) proneness to psychopathology and PTSD, and (h) the dimensions of the self-structure adversely affected by states of posttraumatic shame and guilt. The experience of posttraumatic states of shame and guilt are associated with compounded affective processes in PTSD, depression, and substance use disorders.


Intrarenal infusion of angiotensin-(1-7) modulates renal functional responses to exogenous angiotensin II in the rat.
In the present study we investigated the possible role of angiotensin-(1-7) [Ang-(1-7)] in modulating renal functional responses to intrarenal (i.e.) infusion of angiotensin II (ANG II) in normotensive anesthetized rats. ANG II (6 ng/min, n = 14) decreased glomerular filtration rate (GFR), renal plasma flow (RPF), absolute and fractional sodium excretion by -24 +/- 5, -25 +/- 6, -44 +/- 6 and -28 +/- 7%, respectively (p < 0.05). i.r. infusion of Ang-(1-7) (50 ng/min, n = 13) did not significantly alter GFR (+6 +/- 4%) but reduced RPF by -19 +/- 7% (p < 0.05). Ang-(1-7) increased absolute and fractional sodium excretion by +36 +/- 6 and +37 +/- 8%, respectively (p < 0.05). Infusion of Ang-(1-7) did not prevent the decreases in GFR and RPF but completely blunted the decreases in absolute (-2 +/- 2%) and fractional sodium excretion (-4 +/- 4%) induced by ANG II (n = 11). Blockade of the Ang-(1-7) receptor by [7-D-Ala]-Ang-(1-7) (5 microg/min, n = 11) significantly decreased GFR, RPF, absolute and fractional sodium excretion by -28 +/- 7, -20 +/- 5, -32 +/- 7 and -24 +/- 4%, respectively (p < 0.05), suggesting that the action of endogenous ANG II is unopposed by compensatory effect of endogenous Ang-(1-7). i.r. infusion of Ang-(1-7) (n = 10) did not alter the effect of Ang-(1-7) receptor blockade on RPF (-21 +/- 6%) but blunted its effects on GFR (+4 +/- 3%) and absolute (+7 +/- 5%) and fractional (+6 +/- 4%) urinary sodium excretion probably by displacing the receptor blocker. While exogenous ANG II during blockade of the Ang-(1-7) receptor and the AT(2) receptor (by PD 123319; 1 microg/min i.r., n = 9) resulted in the same decreases in absolute and fractional sodium excretion (-39 +/- 8 and -38 +/- 6%, respectively, p < 0.05) as did ANG II in the absence of Ang-(1-7) receptor blockade. These results suggest that in normotensive rats high i.r. Ang-(1-7) concentration attenuates the tubular, i.e. sodium reabsorptive effect, but not the vascular effect of exogenous i.r. ANG II. Results obtained during blockade of Ang-(1-7) and of AT(2) receptors imply that AT(2) receptors play a role in tubular sodium reabsorption in the presence of high ANG II concentration.


Heart rate and cardiovascular events: a nested case-control in Isfahan Cohort Study.
Elevated heart rate (HR) is known to be a risk factor. The aim of the present study was to investigate the association of HR with the incidence of cardiovascular disease (CVD) in Iranian adults. The Isfahan cohort study (ICS) was a longitudinal study started in 2001 on 6504 adults aged ≥ 35 years in urban and rural areas of central Iran. In a nested case control study, a control was randomly selected for each CVD event occurring during 7 years of follow up using density sampling method. HR at baseline was assessed by electrocardiogram. CVD was defined as incident coronary heart disease (myocardial infarction, unstable angina and sudden cardiac death) and stroke. The odds ratios (OR) were estimated by conditional logistic regression. 432 participants with CVD events in the case group and 401 participants free of CVD in the control group were included in the analysis. While HR did not show any significant relationship with CVD events in the crude model (P = 0.208), it was detrimentally associated with them when age was included (OR = 1.01, 95% CI: 1.00 - 1.02, P = 0.024). A dose response effect of quintiles of HR was seen in which significant association with CVD events started at third quintile [OR = 1.98 (1.15 - 3.41)] and increased toward fifth quintile [OR = 2.53 (1.47 - 4.36)] in the adjusted model for age, sex and HR-lowering drugs (P for trend = 0.001). This association remained statistically significant when other traditional risk factors were included in the model. An elevated heart rate was associated with the occurrence of cardiovascular events. It can be considered as a predictor of cardiovascular disease independently of other risk factors in Iranian adults.


Medical students and drugs: further neuropsychological and use pattern considerations.
Drug use patterns of 134 freshman medical students were surveyed. Ninety-four percent reported experience with alcohol, 72% with marijuana, 24% with hallucinogens or stimulants, and 7% with opiates. Persons who reported use of the least socially sanctioned substances (hallucinogens, stimulants, opiates) were a subgroup of marijuana users, whereas marijuana experienced students were a subgroup of alcohol users. Increasing marijuana use appears to be associated with increased alcohol consumption. Twenty-five students who had never used marijuana and 26 students who had used it on 50 or more occasions were compared on three neuropsychological tests: Tactual Performance Test, Minnesota Perceptuo-Diagnostic Test, and the Hutt Adaptation of the Bender Gestalt Test. There were no neuropsychological differences between marijuana experienced and marijuana naive subjects. The methodological difficulties in studying the long-term cerebral effects of marijuana are discussed.


Probing the specificity of protein-protein interactions by quantitative chemical proteomics.
Chemical proteomics is a versatile tool to investigate protein-small molecule interactions, but can be extended to probe also secondary binding investigating small molecule-protein 1-protein 2 interactions, providing insight into protein scaffolds. This application of chemical proteomics has in particular been applied extensively to cyclic nucleotide (cAMP, cGMP) signaling. cAMP regulates cellular functions primarily by activating cAMP-dependent protein kinase (PKA). Compartmentalization of PKA plays an important role in the specificity of cAMP signaling events and is mediated by interaction of the regulatory subunit (PKA-R) with A-kinase anchoring proteins (AKAPs), which often form the core of even larger protein machineries. The selective binding of AKAPs to one of the major isoforms PKA-R type I (PKA-RI) and PKA-R type II (PKA-RII) is an important feature of cAMP/PKA signaling. However, this specificity is not well established for most AKAPs. Here, we describe a chemical proteomics approach that combines cAMP-based affinity chromatography with quantitative mass spectrometry to investigate PKA-R isoform/AKAP specificity directly in lysates of cells and tissues of any origin. With this tool, several novel PKA-R/AKAP specificities can be easily resolved.


Hematological and biochemical changes in organically farmed sheep naturally infected with Fasciola hepatica.
A naturally occurring outbreak of fasciolosis in a group of 20 Merinolandschaf (German Merino) sheep was studied. Hematological and blood biochemical values in sheep spontaneously infected with liver fluke Fasciola hepatica were compared with equivalent values in 20 parasite-free sheep from organically farmed flock. Investigated animals were kept in outdoor system, on pastures covered with swamps, which remain flooded after rainy season. Significantly lower red blood cell (RBC) count, lymphocytes, hemoglobin (Hb), packed cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatinine, and albumin were recorded in sheep from the infected herd, whereas white blood cell (WBC) count, eosinophil, segmented and band neutrophil count, mean corpuscular volume (MCV), gamma-glutamyl transferase (GGT), concentrations of glucose, and globulins were significantly higher than in the parasite free herd. No significant correlation between the investigated blood parameters and the number of F. hepatica eggs in the feces was detected. This study shows that hematological and biochemical values can be useful in early diagnosis and prognosis of sheep fasciolosis.


Superporous agarose monoliths as mini-reactors in flow injection systems. On-line monitoring of metabolites and intracellular enzymes in microbial cultivation processes.
A new type of agarose material, superporous agarose, was used as a support material in an analytical system designed for monitoring of bioprocesses with respect to metabolites and intracellular enzymes. The superporous agarose was used in the form of miniaturised gel plug columns (15 x 5.0 mM I.D. monolithic gel bed). The gel plugs were designed to have one set of very large pores (about 50 microm in diameter) through which cells, cell debris and other particulate contaminants from the bioreactor could easily pass. The material also had normal diffusion pores (300 A) characteristic of all agarose materials, providing ample surface for covalent attachment of antibodies and enzymes used in the analytical sequence. The superporous agarose gel plug columns were characterised with respect to flow properties and handling of heavy cell loads as well as dispersion of injected samples (a Bodenstein number of about 40 was observed with acetone tracer at a flow rate of 1 ml min(-1)). To evaluate the practical performance of the superporous gel plug columns, two applications were studied: (1) on-line determination of glucose in cultivation broth (gel plug with immobilized glucose oxidase) and (2) immunochemical quantification of intracellular beta-galactosidase in E. coli (gel plug with lysozyme to achieve cell lysis and gel plug with antibodies against beta-galactosidase).


Translocation inhibitors define specificity of protein kinase C isoenzymes in pancreatic beta-cells.
The protein kinase C (PKC) family consists of 11 isoenzymes. Following activation, each isoenzyme translocates and binds to a specific receptor for activated C kinase (RACK) (Mochly-Rosen, D. (1995) Science 268, 247-251) that provides an anchoring site in close proximity to the isoenzyme's specific substrate. Pancreatic islet cells contain at least six PKC isoenzymes (Knutson, K. L., and Hoenig, M. (1994) Endocrinology 135, 881-886). Although PKC activation enhances insulin release, the specific function of each isoenzyme is unknown. Here we show that following stimulation with glucose, alphaPKC and epsilonPKC translocate to the cell's periphery, while deltaPKC and zetaPKC translocate to perinuclear sites. betaC2-4, a peptide derived from the RACK1-binding site in the C2 domain of betaPKC, inhibits translocation of alphaPKC and reduces insulin response to glucose. Likewise, epsilonV1-2, an epsilonPKC-derived peptide containing the site for its specific RACK, inhibits translocation of epsilonPKC and reduces insulin response to glucose. Inhibition of islet-glucose metabolism with mannoheptulose blocks translocation of both alphaPKC and epsilonPKC and diminishes insulin response to glucose while calcium-free buffer inhibits translocation of alphaPKC but not epsilonPKC and lowers insulin response by 50%. These findings illustrate the unique ability of specific translocation inhibitors to elucidate the isoenzyme-specific functions of PKC in complex signal transduction pathways.


Contribution of lactic acid by the ampullary and isthmic parts of rabbit Fallopian tube under altered hormonal conditions.
The capability of the ampullary and isthmic parts of the rabbit Fallopian tube to contribute lactic acid (LA) to the tubal lumen was examined using an in vitro method of study. In addition concomitant study was also made using the uterus. It was found that in the oestrous rabbit, the isthmus contributed LA at a higher rate than that of the ampulla; the uterine contribution was minimal. Ovariectomy reduced while exogenous oestrogen enhanced this ability in all the three tissues. In mated post-ovulatory rabbits the contributory efficiency of either tubal part changed and appeared to exhibit a peak value in the presence of the ovum; the uterus showed no such changes. This is discussed in relation to the ability of the egg to secure its energy source until it attained morula/the pre-blastocyst stage.


Temporal performance in 4-8 year old children. The effect of chronometric information in task execution.
The present study explores temporal performance in children between 4 and 11 years of age. In order to optimise the children's performance, verbal instructions and demonstrations were used to specify the temporal requirement of the task. Moreover, specific feedback was given, which indicated the success of a response, or, in case of a mistake, indicated the type of error made. The task was presented in two phases: training and experimental. In the first phase, the child practised until a learning criterion was reached. In the experimental phase, four conditions were defined according to the information provided to the children. In general, the results of the training phase showed an improvement in performance with advancing age. Significant differences were found in children between 7 and 8 years of age, which points to an important transition period. Results of the experimental phase showed a pronounced impact of instructions on the performance of the task and on the strategies used to control timing behaviour. Specifically, the instructions that contained chronometric information caused an impairment in task performance and a change in the strategy used.


Spectral representation of reduced protein models.
We consider a spectrum-like two-dimensional graphical representation of proteins based on a reduced protein model in which 20 amino acids are grouped into five classes. This particular grouping of amino acids was suggested by Riddle and co-workers in 1997. The graphical representation is based on depicting sequentially the amino acids on five horizontal lines at equal separations. One-letter codes, B, O, U, X and Y, to which numerical values 1 to 5 have been assigned, are suggested as labels for the fictional amino acids that represent all the amino acids within each group. The approach is illustrated on ND6 proteins of eight species having from 168 to 175 amino acids. While visual inspection of the novel spectral graphical representations of proteins may reveal local similarities and dissimilarities of protein sequences, arithmetic manipulations of spectra offer an elegant route to graphic visualization of the degree of similarity for selected pairs of proteins.


Long-term results of the clinical use of an expanded polytetrafluoroethylene surgical membrane as a pericardial substitute.
We report our long-term experience with the clinical use of an expanded polytetrafluoroethylene surgical membrane as a pericardial substitute in 61 children with congenital heart disease. Twenty-three of these children underwent reoperation successfully, and advantages of the membrane in resternotomy and dissection of the pericardial space were recognized. We have so far experienced no major complications from the use of an expanded polytetrafluoroethylene surgical membrane. We conclude that the clinical use of an expanded polytetrafluoroethylene surgical membrane as a pericardial substitute is valuable in children with congenital heart disease who are expected to undergo reoperation in the near future.


Preventing fractures with hip protectors.
Hip protectors are an effective strategy for reducing fall-related hip fractures in long-term care settings. The success of a hip protector program is enhanced by educational activities and administrative support, and a nurse champion to assume responsibility for the hip protector program.


Selective transcription of rat proenkephalin fusion genes from the spermatogenic cell-specific promoter in testis of transgenic mice.
The rat and mouse proenkephalin genes each contains two distinct promoters, one of which is utilized exclusively by spermatogenic cells. The germ cell-specific promoter lacks TATA sequences, is G+C rich, and contains multiple initiation sites. To investigate the nature of the cis-acting elements that determine selective transcription of the proenkephalin gene in male germ cells, two rat proenkephalin-chloramphenicol acetyltransferase fusion genes containing the two different promoter regions as well as 1.6 or 0.3 kilobases, respectively, of 5'-flanking sequence were expressed in transgenic mice. Multiple transgenic lines were developed which expressed the fusion genes in testis, brain, and heart but not in tissues that do not normally express the proenkephalin gene. Fusion gene transcripts in transgenic mouse testes were localized to those spermatogenic cell types that utilize the spermatogenic cell promoter and were selectively and accurately initiated from the multiple rat germ cell start sites. Transgenic mice thus provide a useful model for the localization and characterization of cis-acting elements mediating transcription of the proenkephalin gene from its germ cell-specific promoter.


Indicators for the total duration of premenopausal endogenous estrogen exposure in relation to BMD.
Previous studies have shown that age at menopause is an important indicator of duration of endogenous estrogen exposure. The present study investigates whether combining more information on reproductive factors is useful in estimating individual total duration of exposure to endogenous estrogens. Bone mineral density (BMD) was used as operational outcome. The study population consisted of 3476 white women living in Eindhoven, The Netherlands, aged 46-57 years, either pre- (n = 2420) or postmenopausal (n = 1056). BMD of the lumbar spine was measured by dual X-ray absorptiometry. Information on reproductive factors was obtained with questionnaires. The number of reproductive years explained 4.8% of the variance in BMD, while age at menopause alone accounted for 3.6%. Duration of lactation or oral contraceptive use did not add to the proportion of variance explained. The effect of reproductive years on BMD was stronger in older women. No significant associations with BMD were found for other reproductive variables. The number of miscarriages in premenopausal women (beta = 0.00760, SE = 0.00357, P = 0.03) explained only 0.16% of the variance in BMD. We conclude that it is not necessary to use more reproductive factors besides age at menopause and menarche in determining total duration of endogenous estrogen exposure.


[Bio panning of human stem cell factor(2) mimetic peptides from phage displayed random peptide library].
To screen human stem cell factor (hSCF) mimetic peptides in vitro with a phage-display random peptide library. Phage clones with high hSCF receptor (rc-kit/Ig 1-3)-binding activity was screened from phage-displayed random hepta/dodecapeptide library by phage enzyme-linked immunosorbent assay (ELISA). Phage single DNA was extracted and sequenced. Four kinds of peptide with higher c-Kit/Ig 1-3 binding activity were chosen for synthesis and characterized by using cell proliferation assay with 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) method in UT-7 cells. Eleven Ph.D.-C7C clones and eight Ph.D-12 phage clones with high hSCF receptor-binding activity were selected from phage-displayed random hepta/dodecapeptide library, respectively. Sequence analysis showed there were no homologous sequence between hSCF and these screened mimetic peptides except one homologous sequence DPSPHTH found in heptapeptide library. All these four synthesized peptides (CE3, CE16, LE4, and LE20), particularly CE16 and LE20, stimulated UT-7 cell proliferation. Four hSCF mimetic peptides were successfully isolated from phage-displayed random peptide library..


Nonpharmacologic Interventions may Reduce Mental Distress in Adults Undergoing Dental Treatment.
Non-pharmacological interventions for reducing mental distress in patients undergoing dental procedures: Systematic review and meta-analysis. Burghardt S, Koranyi S, Magnucki G, Strauss B, Rosendahl J. J Dent. 2018;69:22-31. None reported TYPE OF STUDY DESIGN: Meta-analysis of randomized controlled trials.


Intermediate Moral Respect and Proportionality Reasoning.
In a recent article in this journal Jonathan Pugh critiques the idea of intermediate 'moral respect' which some say is owed to embryos. This concept is inherent within the 'principle of proportionality', the principle that destructive research on embryos is permissable only if the research serves an important purpose. Pugh poses two specific questions to proponents of the idea of intermediate moral respect. This article argues that while the questions posed by Pugh are certainly pertinent to the debate, the hypothetical responses he suggests to these questions do not quite get to the core of what is troublesome about the concept. The article suggests alternative responses to Pugh's questions in order to focus attention on more fundamental problems facing the idea of intermediate moral respect, while also pointing to how the intermediate moral respect proponent might best develop these responses. It goes on to argue that these hypothetical responses fail to answer convincingly the questions posed. More specifically, this article challenges two possible justifications for the distinct idea of intermediate moral respect, namely the argument from potentiality (the argument raised by Pugh) and an argument from the proportionality of fundamental moral status (not considered by Pugh). The article also raises a dilemma inherent in the application of the principle of proportionality to cases involving beings to which intermediate moral respect is owed even where it is allowed, ex hypothesi, that both the category of intermediate moral respect and the general proportionality reasoning underpinning the principle of proportionality are basically cogent. This article thus develops and adds to the challenge laid down by Pugh to proponents of the idea of intermediate moral respect.


[Thyroid function in patients with ventricular arrhythmia].
Hyperthyroidism was diagnosed in 4.8% of 21 patients with persistent ventricular arrhythmias (grades 2 to 5 by the Lown-Wolf classification), and latent hypothyroidism was diagnosed in 38.1% of those, using up-to-date radioimmune assays of thyroid hormones and the thyroliberin (TRH) test. It was those conditions that made arrhythmias resistant to antiarrhythmic treatment in most cases. Latent thyroid dysfunction was particularly common in elderly patients with ventricular arrhythmias and aggravated thyroid history or abnormal thyroid scintigraphic findings. Substitution therapy for hypothyroidism was conducive to the disappearance of paroxysms of ventricular tachycardia and the decrease in total number and grades of ventricular extra-systoles in patients with ventricular arrhythmias; moreover, sensitivity to antiarrhythmic agents developed to replace an earlier resistance.


Validity of the GDS-4 revisited.
This article points out several flaws in an earlier article (Chau, Martin, Thompson, Chang, & Woo, 2006). We note that Chau, Martin, Thompson, Chang, and Woo (2006) had misquoted our work on a 4-item version of the geriatric depression scale (GDS), and the work of the research team, which developed the original 30-item and 15-item versions of the scale. Furthermore, their data analytic methods were flawed, and their conclusions were often not supported by the data they presented. On the basis of these observations, we found no evidence against the use of the 4-item version of the GDS.


No meaningful association between suicidal behavior and the use of IL-17A-neutralizing or IL-17RA-blocking agents.
An emerging class of agents blocking IL-17 signaling represents a very promising therapeutic approach. One of these agents, brodalumab, has been associated with an increased risk of suicide behavior. Areas covered: This review sought to provide an overview strictly focused on suicide behavior signals related to the use of IL-17 agents. Data collection regarding this peculiar safety aspect was primarily based on: (i) a revision of safety outcomes belonging to phase II and phase III trials; (ii) a systematic search using the Pubmed Medline database; and (iii) collecting recent data issued as posters or communications in eminent international meetings. Expert opinion: Whilst secukinumab and ixekizumab were not associated with increased signal of suicidal behavior, being recently approved for the treatment of psoriasis by EMA and FDA, brodalumab raised concern because of suicide behavior cases that led to pause momentarily its development program during pre-marketing stage before obtaining the positive recommendation by an FDA advisory panel for its approval. Indeed, a careful re-evaluation of brodalumab safety profile is being performed and no evidence clarified a significant association or a pathogenic mechanism linking brodalumab treatment to the risk of suicidal behavior, suggesting that cases of suicidal behavior accidentally occurred during brodalumab trials.


Route of incorporation of alveolar palmitate and choline into surfactant phosphatidylcholine in rabbits.
Intratracheal injection of 3-day-old rabbits with radioactively labeled palmitic acid and choline results in an 8-10-fold increase in the efficiency of their incorporation into surfactant phosphatidylcholine when compared to the intravenous injection of these precursors. Based on labeling patterns in microsomal, lamellar body and alveolar wash fractions, the incorporation appears to be via normal surfactant synthetic pathways. Intratracheal injection of phospholipid precursors is useful for producing relatively high specific activity natural surfactant.


[Arthritis related to intestinal anastomoses].
Jejuno-colic and jejuno-ileal anastomoses may provoke arthritis. The recently recognized physiopathology is that of arthritis due to immune complexes related to pullulation of Escherichia coli and of Bacilus fragilis. These cases of arthritis, usually sensitive to therapy, have, in some stutborn cases, required the re-establishment of intestinal continuity, which in each case has made the joint phenomena disappear.


Participation of NMDA and kainate receptors of paraventricular nucleus in cardiovascular responses to glutamate receptor agonist.
The nuclei of the hypothalamus have been shown to be involved in central cardiovascular homeostasis. Recent studies suggest that glutamate-containing neurons have an important role in the regulation of central cardiovascular function. We report first on the effects of intracerebrally injected NMDA and non-NMDA receptor ligands on blood pressure and heart rate in conscious Sprague-Dawley rats. In the second part, we describe the effect of blockade of NMDA or kainate receptors in the paraventricular nucleus on glutamate receptor agonist-induced blood pressure responses. Intracerebroventricular injections of L-glutamic acid, NMDA and kainic acid produced increases in mean arterial pressure. Kainic acid produced significant decreases in heart rate. Microinjection of DL-2-amino-5-phosphonopentanoic acid (APV; 25 and 50 nmol), a competitive NMDA receptor antagonist, into the paraventricular nucleus blunted the increases in the mean arterial pressure evoked by intracerebroventricular injections of NMDA (1 nmol), whereas microinjection of dinitroquinoxaline (DNQX; 20, 40 and 80 pmol), which acts as an antagonist at kainate receptors, failed to antagonize the cardiovascular effects of intracerebroventricular kainic acid (10 pmol). Microinjections of NMDA (100 pmol) into the paraventricular nucleus produced pressor responses, but kainic acid (5 and 10 pmol) failed to affect either mean arterial pressure or heart rate. These results suggest participation of the glutamergic system in cardiovascular regulation via NMDA receptors located within the paraventricular nucleus of the hypothalamus in rats.


Plasma levels of vitamin E and lipoperoxide during paediatric anaesthesia.
We measured plasma levels of vitamin E (total tocopherol) and lipoperoxide in seventeen neonates (less than 10 days), twenty infants (1-12 months) and ten children (1-5 years) during anaesthesia. The seventeen neonates were randomly divided into two groups; seven who received 30 mg X kg-1 of alpha-tocopheryl acetate intramuscularly before anaesthesia and ten who did not. The 20 infants were divided into three groups: Group 1: eight infants who did not receive vitamin E; Group 2: six who received 30 mg X kg-1 of alpha-tocopheryl acetate orally for three days before anaesthesia; Group 3: six who received 30 mg X kg-1 of alpha-tocopheryl acetate intramuscularly three hours before anaesthesia. In the neonates who did not receive alpha-tocopheryl acetate, plasma vitamin E and lipoperoxide levels were unchanged following surgery. In Group 1 infants, plasma vitamin E levels decreased (p less than 0.05) and plasma lipoperoxide levels increased (p less than 0.05). In both neonates who received vitamin E and Group 3 infants the mean plasma vitamin E levels increased significantly (p less than 0.05) following surgery. In Group 2 infants, the levels of plasma vitamin E before surgery were high, as compared to the other groups: however, plasma vitamin E levels decreased following surgery. In the children, the plasma vitamin E levels were unchanged, while the plasma lipoperoxide levels decreased significantly (p less than 0.05) during anaesthesia. It is suggested from our studies that plasma vitamin E levels decrease and plasma lipoperoxide levels increase during anaesthesia and surgery in infants; however, those levels are unchanged in neonates.


Effects of cyanide and dissolved oxygen concentration on biological Au recovery.
The number of discarded electric devices containing traces of Au is currently increasing. It is desirable to recover this Au because of its valuable physicochemical properties. Au is usually dissolved with relatively high concentrations of cyanide, which is associated with environmental risk. Chromobacterium violaceum is able to produce and detoxify small amounts of cyanide, and may thus be able to recover Au from discarded electric devices. This study investigated the effects of cyanide and dissolved oxygen concentration on biological Au recovery. Cyanide production by C. violaceum was sufficient to dissolve Au, while maintaining a high cyanide concentration did not enhance Au dissolution. Increased oxygen concentration enhanced Au dissolution from 0.04 to 0.16 mmol/l within the test period of 70 h. Electrochemical measurement clarified this phenomenon; the rest potential of Au in the cyanide solution produced by C. violaceum increased from -400 to -200 mV, while in the sterile cyanide solution, it was constant in cyanide concentrations ranging from 0 to 1.5 mmol/l and increased in dissolved oxygen concentrations ranging from 0 to 0.25 mmol/l. Therefore, it was clarified that dissolved oxygen concentration is the main factor affecting the efficiency of cyanide leaching of gold by using bacteria.


[Prevalence of positive gonococcal and chlamydial findings in women applying for termination of pregnancy. A screening evaluation].
All 335 pregnant women referred for termination of pregnancy in the first trimester were examined preoperatively for infection with Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) and were questioned about symptoms of gynaecological infection. Only 0.6% had positive cultures for gonorrhoeae while CT infection was demonstrated in 7.8%. Half of the women with infection had one of the following symptoms: discharge, pain or dysuria. These same symptoms were, however, also found in half of the women without infection. All gynaecological special departments in Denmark and a few of the surgical departments which carry out termination of pregnancy replied to a questionnaire about routine examinations for NG and CT in cases of termination of pregnancy. Only 29.2% of the departments made routine preoperative examinations for CT, NG or both. It is concluded that there are no indications for routine examination for gonococcal infection whereas screening for CT infection appears to be indicated.


Electrophoretic variation of avian rotavirus RNA in polyacrylamide gels.
Five different sorts of RNA profiles (electropherogroups 1 to 5) were recognised when avian rotavirus RNAs were electrophoresed in polyacrylamide gels. These could be distinguished from the RNAs of mammalian group A, B and C rotaviruses. Viruses belonging to electropherogroups 1, 2/3 and 4 were detected in chickens and viruses belonging to electropherogroups 1, 2, 3 and 5 were detected in turkeys. A minimum of three electropherogroups was recognised in each of four longitudinal surveys of broiler chicken flocks. On examination of rotaviruses from different clinical outbreaks of diarrhoea and from different longitudinal surveys minor differences involving almost all the RNA segments were observed when rotaviruses from the same electropherogroup were compared. Detailed RNA analysis in two of the surveys detected up to three electropherotypes within each electropherogroup.