Reduction of the local toxicity of intraperitoneal chemotherapy; an experimental model.
Intraperitoneal (I.P.) administration of chemotherapeutic agents, in particular, cisplatin and bleomycin is successfully used in the treatment of ovarian and gastrointestinal malignancies. Their use however, is limited by the rapid development of adhesions. The vinca alkaloid group of chemotherapeutic agents has been shown to impair the acute inflammatory reaction, which plays an important role in adhesion formation, thereby providing the rationale for addition of vindesine to an I.P. regimen containing cisplatin and bleomycin. A rat model was used to study the effects of various I.P. chemotherapeutic regimens on adhesion formation. Four groups were studied (30 rats/Group). Regimens used included Group A (Saline), Group B (Vindesine 0.1 mg/kg), Group C (Cisplatin 5 mg/kg and Bleomycin 0.2 mg/kg), and Group D (Cisplatin 5 mg/kg, Bleomycin 0.2 mg/kg and Vindesine 0.1 mg/kg). In order to grade adhesion formation, animals were sacrificed on days 2, 14 and 28, with the severity of adhesions graded on a scale of I-IV in order of increasing severity. Adhesion formation in Group A (Saline) and Group B (Vindesine) was similar. The addition of Vindesine to Group C (Cisplatin and Bleomycin) resulted in a significant reduction in adhesion formation (p less than 0.01). The results of this experimental study suggest that the local toxicity of I.P. chemotherapy may be reduced by the inclusion of a vinca alkaloid.


The association of macro- and microstructure of the corpus callosum and language lateralisation.
The present study aimed to examine how differences in functional lateralisation of language are related to interindividual variations in interhemispheric connectivity. Utilising an fMRI silent word-generation paradigm, 89 left- and right-handed subjects were subdivided into four lateralisation subgroups. Applying morphological and diffusion-tensor MRI, midsagittal cross-sectional area as well as quantitative measures of molecular diffusion (anisotropy, mean diffusion) of the corpus callosum were determined to assess interhemispheric connectivity. Statistical analyses revealed group differences in molecular diffusion but not in callosal size, which may be interpreted to reflect a stronger and/or faster interhemispheric connection in strongly left-lateralised subjects as compared to moderately left-lateralised, bilateral, or moderately right-lateralised subjects.


Human estrogen receptor α antagonists, part 2: Synthesis driven by rational design, in vitro antiproliferative, and in vivo anticancer evaluation of innovative coumarin-related antiestrogens as breast cancer suppressants.
New twelve in silico designed coumarin-based ERα antagonists, namely 3DQ-1a to 3DQ-1е, were synthesized and confirmed as selective ERα antagonists, showing potencies ranging from single-digit nanomolar to picomolar. The hits were confirmed as selective estrogen receptor modulators and validated as antiproliferative agents using MCF-7 breast cancer cell lines exerting from picomolar to low nanomolar potency, at the same time showing no agonistic activity within endometrial cell lines. Their mechanism of action was inspected and revealed to be through the inhibition of the Raf-1/MAPK/ERK signal transduction pathway, preventing hormone-mediated gene expression on either genomic direct or genomic indirect level, and stopping the MCF-7 cells proliferation at G0/G1 phase. In vivo experiments, by means of the per os administration to female Wistar rats with pre-induced breast cancer, distinguished six derivatives, 3DQ-4a, 3DQ-2a, 3DQ-1a, 3DQ-1b, 3DQ-2b, and 3DQ-3b, showing remarkable potency as tumor suppressors endowed with optimal pharmacokinetic profiles and no significant histopathological profiles. The presented data indicate the new compounds as potential candidates to be submitted in clinical trials for breast cancer therapy.


Statins prevent NF-kappaB transactivation independently of the IKK-pathway in human endothelial cells.
Statins have been linked to a wide range of vascular benefits, many of them are likely to be due to attenuation of chronic vascular inflammation. Nuclear factor kappaB (NF-kappaB) is one of the key regulators of transcription of a variety of genes involved in immune and inflammatory responses. Therefore, we investigated the effect of statins on TNF-alpha-induced NF-kappaB signaling in human endothelial cells (EC). ECs were pre-incubated for 16 h with cerivastatin (10(-9) to 10(-7) M) or vehicle in the presence or absence of mevalonate, followed by stimulation with 20 ng/ml TNF-alpha. Statin-treatment prevented TNF-alpha-induced NF-kappaB binding activity, nuclear translocation of the NF-kappaB p65 subunit, as well as NF-kappaB controlled tissue factor (TF) gene transcription in cultured EC. IkappaBalpha phosphorylation and IkappaBalpha degradation, however, still occurred in statin-treated cells. TNF-alpha also activated phosphatidylinositol (PI)3-kinase, as reflected by phosphorylation of Akt. Statin treatment of cells abrogated TNF-alpha-induced Akt phosphorylation and p65 nuclear translocation. As observed with statins, inhibition of PI3-kinase activity by Ly294002 also blocked TNF-alpha-induced p65 translocation, but did not prevent IkappaBalpha phosphorylation nor IkappaBalpha degradation. These studies demonstrate that TNF-alpha-induced NF-kappaB activation is abrogated by statin treatment in HUVEC independently of the classical IKK-pathway but via inhibition of PI3-kinase/Akt signaling.


Rheology: liquefaction of quicksand under stress.
People or animals caught in quicksand find it very hard to escape. Here we show that quicksand acts as a trap because it becomes unstable when it is forced to move--first it liquefies, and then it collapses. But a simple sinking test demonstrates that it is impossible for a human to be drawn into quicksand altogether.


Transverse Cervical Artery Pseudoaneurysm: An Unusual Delayed Complication of Radical Neck Dissection.
Pseudoaneurysm formation in the transverse cervical artery, post radical neck dissection, leading to massive hemorrhage, is a rare but life threatening occurrence. We report a patient with pseudoaneurysm of transverse cervical artery, post salvage radical neck dissection, presenting with recurrent and significant hemorrhage after 3 weeks of surgery. A pseudoaneurysm involving transverse cervical artery was revealed by digital subtraction angiography and treated by endovascular coil embolization.


Overlapping gene coexpression patterns in human medullary thymic epithelial cells generate self-antigen diversity.
Promiscuous expression of numerous tissue-restricted self-antigens (TRAs) in medullary thymic epithelial cells (mTECs) is essential to safeguard self-tolerance. A distinct feature of promiscuous gene expression is its mosaic pattern (i.e., at a given time, each self-antigen is expressed only in 1-3% of mTECs). How this mosaic pattern is generated at the single-cell level is currently not understood. Here, we show that subsets of human mTECs expressing a particular TRA coexpress distinct sets of genes. We identified three coexpression groups comprising overlapping and complementary gene sets, which preferentially mapped to certain chromosomes and intrachromosomal gene clusters. Coexpressed gene loci tended to colocalize to the same nuclear subdomain. The TRA subsets aligned along progressive differentiation stages within the mature mTEC subset and, in vitro, interconverted along this sequence. Our data suggest that single mTECs shift through distinct gene pools, thus scanning a sizeable fraction of the overall repertoire of promiscuously expressed self-antigens. These findings have implications for the temporal and spatial (re)presentation of self-antigens in the medulla in the context of tolerance induction.


Prevalence of residual B-cell function related to age at onset and genetic profile in newly diagnosed type I diabetics.
Patients with type I (insulin-dependent) diabetes mellitus maintain B-cell function for a varying period of time after onset. This is commonly held to account for post-initial remission. To estimate residual B-cell function we measured plasma and 24-h urinary C-peptide in 68 type I diabetic patients (age range 4-35 years, within 10-180 days of the onset of symptoms, typed for HLA-A, -B, -C and DR loci. A positive correlation (r = 0.26; p less than 0.05) was found between urinary C-peptide levels and the age of the patient. The analysis of variance of urinary C-peptide values on the basis of the presence or absence of DR3 and DR4 antigens revealed that the DR3-positive patients had reduced excretion (15.2 +/- 9.2 SD micrograms/24h) with respect to the others (22.7 +/- 15.5 SD micrograms/24h) (F = 6.35; p less than 0.05). No interaction effect was found in DR3/4 positive patients. Hence, late onset patients appear to have higher residual C-peptide secretion. In the light of these findings, the assessment of B-cell function and genetic profile may be useful in predicting which patients are likely to have remission periods and identifying the metabolic consequences of even minimal endogenous insulin secretion.


On-treatment platelet reactivity through the thromboxane A2 or P2Y12 platelet receptor pathways is not affected by pelacarsen.
Pelacarsen decreases plasma levels of lipoprotein(a) [Lp(a)] and oxidized phospholipids (OxPL). It was previously reported that pelacarsen does not affect the platelet count. We now report the effect of pelacarsen on on-treatment platelet reactivity. Subjects with established cardiovascular disease and screening Lp(a) levels ≥60 mg per deciliter (~ ≥150 nmol/L) were randomized to receive pelacarsen (20, 40, or 60 mg every 4 weeks; 20 mg every 2 weeks; or 20 mg every week), or placebo for 6-12 months. Aspirin Reaction Units (ARU) and P2Y12 Reaction Units (PRU) were measured at baseline and the primary analysis timepoint (PAT) at 6 months. Of the 286 subjects randomized, 275 had either an ARU or PRU test, 159 (57.8%) were on aspirin alone and 94 (34.2%) subjects were on dual anti-platelet therapy. As expected, the baseline ARU and PRU were suppressed in subjects on aspirin or on dual anti-platelet therapy, respectively. There were no significant differences in baseline ARU in the aspirin groups or in PRU in the dual anti-platelet groups. At the PAT there were no statistically significant differences in ARU in subjects on aspirin or PRU in subjects on dual anti-platelet therapy among any of the pelacarsen groups compared to the pooled placebo group (p > 0.05 for all comparisons). Pelacarsen does not modify on-treatment platelet reactivity through the thromboxane A2 or P2Y12 platelet receptor pathways.


Indication of metal homeostasis disturbance in earthworm Eisenia fetida after exposure to semi-solid depot sludge.
Treated sewage sludge is commonly used in agriculture as fertilizer. It is, therefore, necessary to determine possible detrimental influences of sludge application on soil contamination and accumulation of contaminants in tissues of terrestrial animals, which in the long run could also have undesirable effects on humans. With that aim, the study was performed using earthworm Eisenia fetida as test organism and semi-solid depot sludge from a wastewater treatment plant as exposure media. The concentrations of 26 metals/metalloids were determined in depot sludge, and their bioaccumulation was estimated in whole tissue of E. fetida, and for the first time in the soluble tissue fraction, which represents metal fraction available for metabolic requirements and toxic effects. Obtained results have revealed acceptable levels of several elements (Cd, Cr, Cu, Ni, Pb, Zn) in depot sludge, when compared to currently valid regulations, and only moderate accumulation of some elements (e.g. As, Ba, Cd, Co, Fe, Tl, V, and Zn) in earthworms, as a consequence of exposure to depot sludge. However, a concentration increase after exposure to depot sludge was observed in E. fetida for several elements (Cd, Mo, and Zn), which were present in lower concentrations in the exposure mixtures than in soil. Contrary, a concentration decrease was observed for Cs, Mn, and Rb, although they were present in higher concentrations in depot sludge than in soil. It was an indication of disturbance in metal homeostasis in earthworms, possibly caused by exposure to complex mixture of contaminants present in depot sludge. The cumulative effect of exposure to a number of various contaminants (inorganic, organic, microbiological and pharmaceutical), even if each of them was not present in very high concentrations, could have caused distress in earthworms exposed to depot sludge.


Indopolystoma n. gen. (Monogenea, Polystomatidae) with the description of three new species and reassignment of eight known Polystoma species from Asian frogs (Anura, Rhacophoridae).
The polystomes (Monogenea, Polystomatidae) radiated across semi-aquatic tetrapods including all three amphibian orders, freshwater turtles and the hippopotamus. Prior to this study, phylogenetic analyses revealed that the most diverse and widespread genus, Polystoma, was not monophyletic; a lineage comprising four undescribed species from the bladder of Zhangixalus spp. (Rhacophoridae) in Asia occupied a deep phylogenetic position. Regarding vicariance biogeography and molecular dating, the origin of this lineage is correlated with the breakup of Gondwanaland in the Mesozoic period. Based on a Bayesian analysis of four concatenated genes (18S, 28S, COI and 12S) and morphological evidence, one new genus, Indopolystoma n. gen., and three new species, sampled in Japan and China, are described here: Indopolystoma viridi n. sp. from Z. viridis of Japan, Indopolystoma elongatum n. sp. from Z. arboreus of Japan, and Indopolystoma parvum n. sp. from Z. omeimontis of China. Indopolystoma is unique amongst polystome genera infecting anurans by possessing a small haptor relative to the body size, posteriormost marginal hooklet C1 much bigger than hooklets C2-C8 with conspicuous broad blade and guard and a pair of hamuli lacking a deep notch. Eight species of Asian Polystoma, all from rhacophorids, are transferred as Indopolystoma carvirostris (Fan, Li & He, 2008) n. comb., I. hakgalense (Crusz & Ching, 1975) n. comb., I. indicum (Diengdoh & Tandon, 1991) n. comb., I. leucomystax (Zhang & Long, 1987) n. comb., I. mutus (Meng, Song & Ding, 2010) n. comb., I. pingbianensis (Fan, Wang & Li, 2004) n. comb., I. rhacophori (Yamaguti, 1936) n. comb., and I. zuoi (Shen, Wang & Fan, 2013) n. comb.


Specific formation of beads-on-a-chain structures on giant DNA using a designed polyamine derivative.
Fluorescence microscopy was used to study the folding transition of giant DNAs, T4 DNA (ca. 166 kbp), and lambda DNA (ca. 48 kbp), which proceeds through intermediates with intramolecular segregation induced by pteridine-polyamine conjugates, i.e., 2-amino-6,7-dimethyl-4-(4,9,13-triazatridecylamino)pteridine and -4-(3-(aminopropyl)amino)pteridine. According to the results of DNA denaturation, UV and fluorescent spectroscopy, and transmission electron microscopic observations, it became clear that DNA folding induced by the polyamine derivative is not a continuous shrinking process but a combination of discontinuous processes.


Colchicine-like effect of diethylstilbestrol (DES) on mammalian cells in vitro.
Diethylstilbestrol (DES), a synthetic estrogen, showed colchicine-like effects in vitro on cells of the cell lines such as Chinese hamster fibroblast of thymus origin (CHT), rat liver (DL), rat erythroblastic leukemia (EDEN-1/TC) and HeLa-S3. Metaphase arrest was induced 3 h after treatment with 15 microgram/ml of DES and polyploid or polynucleated cells were prominently observed more than 24 h after treatment. The arrest, however, was reversible when the agent was removed from the medium. Tetraploid karotypes induced by DES in CHT cells consisted of all double sets of diploid chromosome constitution except one chromosome marker. By clonal selection, several hypotetraploid sublines were successfully isolated from a CHT cell population after the treatment with 15 microgram/ml of DES for 48 h. Some comparative studies of cytological effects of DES with those induced by colcemid indicated that the DES effect was also a mitotic inhibition similar to colchicine.


Maternal alcohol consumption and risk of offspring with congenital malformation: the Japan Environment and Children's Study.
The association between fetal exposure to alcohol and congenital structural disorders remains inconclusive. The present study searched for relationships between maternal alcohol consumption during pregnancy and the risk of congenital malformations. We evaluated the fixed dataset of a large national birth cohort study including 73,595 mothers with a singleton live birth. Information regarding the alcohol consumption of mothers was obtained from self-reported questionnaires. Physicians assessed for 6 major congenital malformations (congenital heart defects [CHDs], male genital abnormalities, limb defects, cleft lip and/or cleft palate [orofacial clefts (OFC)], severe brain abnormalities, and gastrointestinal obstructions) up to 1 month after birth. Multiple logistic regression analysis was performed to identify associations between maternal alcohol consumption during pregnancy and each malformation. The prevalence of maternal drinking in early pregnancy and until the second/third trimester was 46.6% and 2.8%, respectively. The onset of CHD was inversely associated with mothers who quit drinking during early pregnancy (OR 0.85, 95% CI 0.74-0.98). There was no remarkable impact of maternal drinking habit status on the other congenital malformations after adjustment for covariates. Maternal alcohol consumption during pregnancy, even in early pregnancy, displayed no significant adverse impact on congenital malformations of interest. This large-scale Japanese cohort study revealed that no teratogenic associations were found between maternal retrospective reports of periconceptional alcohol consumption and congenital malformations after adjustment for covariates. This is the first nationwide birth cohort study in Japan to assess the effect of maternal alcohol consumption during pregnancy on major congenital malformations. Our finding indicated that maternal low-to-moderate alcohol consumption during pregnancy, even in early pregnancy, displayed no significant adverse impact on congenital heart defects, male genital abnormalities, limb defects, orofacial clefts, severe brain abnormalities, or gastrointestinal obstructions.


Winter activity of Phlebotomus (Transphlebotomus) mascittii, Grassi 1908 (Diptera: Psychodidae) on the island of Corsica.
In the first week of February 2008 on the island of Corsica, 55 Phlebotomus (Transphlebotomus) mascittii, 39 males and 16 females, were caught in a railway tunnel. This is the first proof of the winter activity of the adult P. mascittii in Europe.


LDL receptor-related protein (LRP) in Alzheimer's disease: towards a unified theory of pathogenesis.
To date, mutations in three genes, beta-amyloid precursor protein (APP), presenilin 1 (PS1), and presenilin 2 (PS2), have been found to be causally related to familial Alzheimer's disease (AD). In addition, polymorphisms in three other genes (among others), apolipoprotein E (apoE), alpha2-macroglobulin (alpham), and the low density lipoprotein receptor-related protein (LRP), are implicated to contribute to AD pathogenesis. Interestingly, the encoded gene products are all functionally related in various ways to LRP. Specifically apoE, alpha2m, secreted APP, and amyloid beta-protein (Abeta) complexed to either apoE or alpha2m are ligands of LRP. Furthermore, over-expression of presenilin 1 results in decreased expression of LRP. Since levels of many LRP ligands are increased in Alzheimer's disease and LRP and its ligands are present in senile plaques, decreased LRP function may be a central component in AD pathogenesis. This review explores the current knowledge of LRP in AD and its relationship to the other known AD susceptibility markers.


From Large Deviations to Semidistances of Transport and Mixing: Coherence Analysis for Finite Lagrangian Data.
One way to analyze complicated non-autonomous flows is through trying to understand their transport behavior. In a quantitative, set-oriented approach to transport and mixing, finite time coherent sets play an important role. These are time-parametrized families of sets with unlikely transport to and from their surroundings under small or vanishing random perturbations of the dynamics. Here we propose, as a measure of transport and mixing for purely advective (i.e., deterministic) flows, (semi)distances that arise under vanishing perturbations in the sense of large deviations. Analogously, for given finite Lagrangian trajectory data we derive a discrete-time-and-space semidistance that comes from the "best" approximation of the randomly perturbed process conditioned on this limited information of the deterministic flow. It can be computed as shortest path in a graph with time-dependent weights. Furthermore, we argue that coherent sets are regions of maximal farness in terms of transport and mixing, and hence they occur as extremal regions on a spanning structure of the state space under this semidistance-in fact, under any distance measure arising from the physical notion of transport. Based on this notion, we develop a tool to analyze the state space (or the finite trajectory data at hand) and identify coherent regions. We validate our approach on idealized prototypical examples and well-studied standard cases.


Cannabinoids: possible role in patho-physiology and therapy of Gilles de la Tourette syndrome.
High densities of cannabinoid receptors were found in the basal ganglia and hippocampus, indicating a putative functional role of cannabinoids in movement and behaviour. Anecdotal reports suggested beneficial effects of marijuana in Tourette's syndrome (TS). We therefore interviewed 64 TS patients with regard to use of marijuana and its influence on TS symptomatology. Of 17 patients (27%) who reported prior use of marijuana, 14 subjects (82%) experienced a reduction or complete remission of motor and vocal tics and an amelioration of premonitory urges and obsessive-compulsive symptoms. Our results provide more evidence that marijuana improves tics and behavioural disorders in TS. It can be speculated that cannabinoids might act through specific receptors, and that the cannabinoid system might play a major role in TS pathology.


Pattern design of a liquid metal-based wearable heater for constant heat generation under biaxial strain.
As the wearable heater is increasingly popular due to its versatile applications, there is a growing need to improve the tensile stability of the wearable heater. However, maintaining the stability and precise control of heating in resistive heaters for wearable electronics remains challenging due to multiaxial dynamic deformation with human motion. Here, we propose a pattern study for a circuit control system without complex structure or deep learning of the liquid metal (LM)-based wearable heater. The LM direct ink writing (DIW) method was used to fabricate the wearable heaters in various designs. Through the study about the pattern, the significance of input power per unit area for steady average temperature with tension was proven, and the directionality of the pattern was shown to be a factor that makes feedback control difficult due to the difference in resistance change according to strain direction. For this issue, a wearable heater with the same minimal resistance change regardless of the tension direction was developed using Peano curves and sinuous pattern structure. Lastly, by attaching to a human body model, the wearable heater with the circuit control system shows stable heating (52.64°C, with a standard deviation of 0.91°C) in actual motion.


The gatekeeper in vision care. An analysis of the co-ordination of professional services in The Netherlands.
Data from a national survey were used to explore the position of ophthalmologists, general practitioners, orthoptists, optometrists and opticians in the domain of vision care services. Options for organising the gatekeeper function were analysed. This was done on the basis of six cases that the five key occupations considered as their overlapping areas. Nearly all respondents reported to be consulted by patients with the given complaints, indicating rather unclear boundaries between the professions. Further, the opinions indicated preference for a medical gatekeeper (ophthalmologist, GP) rather than a non-medical one (optometrist). Lack of agreement on suggested gatekeeper options suggest other options to consider, like regional networks of GPs, ophthalmologists, orthoptists and optometrists who share the responsibility for a specified client population. At present, such innovative arrangements are being introduced. GPs and optometrists could share the responsibility for gate-keeping and for referring patients to more specialised services.


Pathophysiology of Alzheimer's Disease.
Alzheimer's disease (AD) is the most common neurodegenerative disease leading to dementia worldwide. While neuritic plaques consisting of aggregated amyloid-beta proteins and neurofibrillary tangles of accumulated tau proteins represent the pathophysiologic hallmarks of AD, numerous processes likely interact with risk and protective factors and one's culture to produce the cognitive loss, neuropsychiatric symptoms, and functional impairments that characterize AD dementia. Recent biomarker and neuroimaging research has revealed how the pathophysiology of AD may lead to symptoms, and as the pathophysiology of AD gains clarity, more potential treatments are emerging that aim to modify the disease and relieve its burden.


Phosphorylation of serine 1137/1138 of mouse insulin receptor substrate (IRS) 2 regulates cAMP-dependent binding to 14-3-3 proteins and IRS2 protein degradation.
Insulin receptor substrate (IRS) 2 as intermediate docking platform transduces the insulin/IGF-1 (insulin like growth factor 1) signal to intracellular effector molecules that regulate glucose homeostasis, β-cell growth, and survival. Previously, IRS2 has been identified as a 14-3-3 interaction protein. 14-3-3 proteins can bind their target proteins via phosphorylated serine/threonine residues located within distinct motifs. In this study the binding of 14-3-3 to IRS2 upon stimulation with forskolin or the cAMP analog 8-(4-chlorophenylthio)-cAMP was demonstrated in HEK293 cells. Binding was reduced with PKA inhibitors H89 or Rp-8-Br-cAMPS. Phosphorylation of IRS2 on PKA consensus motifs was induced by forskolin and the PKA activator N(6)-Phe-cAMP and prevented by both PKA inhibitors. The amino acid region after position 952 on IRS2 was identified as the 14-3-3 binding region by GST-14-3-3 pulldown assays. Mass spectrometric analysis revealed serine 1137 and serine 1138 as cAMP-dependent, potential PKA phosphorylation sites. Mutation of serine 1137/1138 to alanine strongly reduced the cAMP-dependent 14-3-3 binding. Application of cycloheximide revealed that forskolin enhanced IRS2 protein stability in HEK293 cells stably expressing IRS2 as well as in primary hepatocytes. Stimulation with forskolin did not increase protein stability either in the presence of a 14-3-3 antagonist or in the double 1137/1138 alanine mutant. Thus the reduced IRS2 protein degradation was dependent on the interaction with 14-3-3 proteins and the presence of serine 1137/1138. We present serine 1137/1138 as novel cAMP-dependent phosphorylation sites on IRS2 and show their importance in 14-3-3 binding and IRS2 protein stability.


[Morbidity following simultaneous pancreas/kidney transplantation].
Simultaneous pancreas-kidney transplantation (SPK) is still associated with the highest rate of morbidity among solid organ transplantations. Although improved long-term survival following SPK has been proven in IDDM patients, a further decrease in morbidity would be desirable. A retrospective, single-center study was performed to investigate the morbidity following SPK and to compare the results to kidney transplantation alone (KTA). Parameters included the rates of relaparotomies, septic complications (urinary tract infection, wound infection, pneumonia), and graft function. Between September 2000 and August 2001, 99 patients underwent transplantation (34 SPK, 63 KTA, 2 pancreas transplants alone). Relaparotomies were performed in six SPK patients (17.6%), mostly due to complications related to the pancreatic graft (n=5). Three reoperations (4.8%) were necessary in KTA patients (p=0.085). Septic complications occurred more often in SPK than in KTA patients (55.9% vs 30.2%, p<0.05). This difference resulted from the high rate of wound infections in SPK patients (35.3%). No intra-abdominal infection or sepsis occurred in any patient. There was one hospital death in SPK and KTA patients, respectively. The rejection rate was similar in SPK (17.6%) and KTA (12.7%, p=0.72). At discharge 91.2% of SPK patients were insulin free and 97.1% free of dialysis. At discharge 96.8% of KTA patients were free of dialysis. SPK is still associated with a higher morbidity (relaparotomies, septic complications) than KTA, although life-threatening complications were rare. There was no increased mortality following SPK.


Assessment and characterization of time-related differences in plaque composition by intravascular ultrasound-derived radiofrequency analysis in heart transplant recipients.
The survival of heart transplant patients is limited by cardiac allograft vasculopathy (CAV). Intravascular ultrasound (IVUS) and IVUS-derived radiofrequency plaque composition analysis (IVUS-RF) provide further information about the process of coronary atherosclerosis. In this study we aimed to assess the time-dependent differences in disease progression in patients with CAV. Fifty-six patients were divided into three groups according to time interval after transplantation (Group I: 1 to 3 months, 18 patients; Group II, 1 to 5 years, 20 patients; Group III: 5 to 15 years, 18 patients). IVUS-RF revealed time-dependent increases in all plaque components. The largest increase was shown for fibrotic, fibrofatty and necrotic tissue between Groups I and II. Dense calcium area increased uniformly in all groups. IVUS-RF-derived plaque type analysis revealed predominantly fibrotic plaques in all groups with a decrease of frequency over time. Fibrolipidic and fibrotic-calcific plaques increased uniformly. High-risk lesions, such as thick-cap fibroatheromas (FAs), increased in Groups I and II and decreased in Group III. Thin-cap FAs were detected only in Group III. IVUS-RF, as compared with gray-scale IVUS, provides better detailed information about the development of CAV by plaque morphology and composition analysis in different stages after heart transplantation. Serial IVUS-RF analysis in these patients may improve the stratification of heart transplant recipients.


Effects of anti-platelet glycoprotein Ib and/or IIb/IIIa autoantibodies on the size of megakaryocytes in patients with immune thrombocytopenia.
To determine whether anti-platelet autoantibodies react with megakaryocytes, as well as with platelets, in immune thrombocytopenia (ITP), 38 ITP patients were studied. They were classified into four groups; anti-platelet glycoprotein Ib-positive (group A, n = 5), anti-platelet glycoprotein II/b/IIIa-positive (group B, n = 2), positive to both antibodies (group C, n = 3), and negative to both antibodies (group D, n = 28). The number and size of megakaryocytes in each group were compared. The number of megakaryocytes in groups A, B, C, and D was 12.8 +/- 8.9, 75.2, 29.1, and 17.0 +/- 21.7/mm2, respectively. The mean cytoplasmic area of megakaryocytes in groups A, B, C, and D was 1001 +/- 26.3, 1621, 1109, and 1311 +/- 235.6/micrograms2, respectively. This finding indicated that, in the presence of anti-platelet glycoprotein Ib, megakaryocytes were not increased in number and were small in size, whereas, in the presence of anti-platelet glycoprotein IIb/IIIa, megakaryocytes were increased in number and in cytoplasmic area. Our study suggested that anti-platelet glycoprotein Ib may impair platelet production by megakaryocytes in ITP.


Selected aspects of the human gut microbiota.
The gut microbiota represents a highly complex assembly of microbes, which interact with each other and with their host. These interactions have various implications in terms of health and disease, and this multi-author review issue will address a number of selected aspects pertaining to gut microbiota research.


Cloning of the rat insulin- like growth factor binding protein-5 gene and DNA sequence analysis of its promoter region.
To understand the regulation of the insulin-like growth factor binding protein-5 (IGFBP-5) gene expression, we have cloned the IGFBP-5 gene from rat genomic libraries and determined its genomic organization as well as the DNA sequence at the 5' flanking region of the gene. The rat IGFBP-5 gene spans at least 17 kilobases (kb) of the genome and contains 4 exons interrupted by 3 introns of approximately 10, 0.6 and 0.7 kb, respectively. Southern blot analysis of the rat chromosome DNA revealed a single copy gene for IGFBP-5 in the haploid genome. Primer extension experiments demonstrated a single transcriptional start site located at 772 nucleotides 5' of the ATG translational start codon. In addition to a TATA box and a CAAT box, multiple putative cis-regulatory elements, including an AP-1, an AP-2 and a binding site for progesterone receptor are present in the promoter region. This finding suggests that the IGFBP-5 gene is one of those that are expressed in a tissue-specific manner.


Modelling estimates of age-specific influenza-related hospitalisation and mortality in the United Kingdom.
Influenza is rarely confirmed with laboratory testing and accurate assessment of the overall burden of influenza is difficult. We used statistical modelling methods to generate updated, granular estimates of the number/rate of influenza-attributable hospitalisations and deaths in the United Kingdom. Such data are needed on a continuing basis to inform on cost-benefit analyses of treatment interventions, including vaccination. Weekly age specific data on hospital admissions (1997-2009) and on deaths (1997-2009) were obtained from national databases. Virology reports (1996-2009) of influenza and respiratory syncytial virus detections were provided by Public Health England. We used an expanded set of ICD-codes to estimate the burden of illness attributable to influenza which we refer to as 'respiratory disease broadly defined'. These codes were chosen to optimise the balance between sensitivity and specificity. A multiple linear regression model controlled for respiratory syncytial virus circulation, with stratification by age and the presence of comorbid risk status (conditions associated with severe influenza outcomes). In the United Kingdom there were 28,516 hospitalisations and 7163 deaths estimated to be attributable to influenza respiratory disease in a mean season, with marked variability between seasons. The highest incidence rates of influenza-attributable hospitalisations and deaths were observed in adults aged 75+ years (252/100,000 and 131/100,000 population, respectively). Influenza B hospitalisations were highest among 5-17 year olds (12/100,000 population). Of all estimated influenza respiratory deaths in 75+ year olds, 50 % occurred out of hospital, and 25 % in 50-64 year olds. Rates of hospitalisations and death due to influenza-attributable respiratory disease were increased in adults identified as at-risk. Our study points to a substantial but highly variable seasonal influenza burden in all age groups, particularly affecting 75+ year olds. Effective influenza prevention or early intervention with anti-viral treatment in this age group may substantially impact the disease burden and associated healthcare costs. The high burden of influenza B hospitalisation among 5-17 year olds supports current United Kingdom vaccine policy to extend quadrivalent seasonal influenza vaccination to this age group. ClinicalTrial.gov, NCT01520935.


The origin and extent of direct amygdala projections to the region of the dorsal motor nucleus of the vagus and the nucleus of the solitary tract.
The site of origin of a direct projection from the amygdala to autonomic regulatory nuclei in the dorsomedial medulla has been precisely localized and delimited in the rabbit by means of the retrograde axonal transport of horseradish peroxidase. Labeled cells in the amygdala were found to be confined within regions of the ipsilateral central nucleus. Many labeled cells were also observed to extend beyond the central nucleus into closely related regions of the substantia innominata and bed nucleus of the stria terminalis.


Higher visceral to subcutaneous fat ratio is associated with small intestinal bacterial overgrowth.
There is a lack of studies evaluating the association between small intestinal bacterial overgrowth (SIBO) and abdominal fat. The aim of this study was to evaluate whether visceral fat area (VFA), subcutaneous fat area (SFA) or visceral to subcutaneous fat ratio (VFA/SFA ratio) were associated with SIBO. In this case-control study, 152 eligible patients submitted to glucose hydrogen/methane breath test who also had computed tomography (CT) of the abdomen performed were included. Clinical and demographic information was obtained. VFA and SFA were measured using Image J software at lumbar 3 level on CT cross-sectional image of the 152 patients included in this study, 68 patients (44.7%) tested positive for SIBO. In the univariate analysis, the presence of SIBO was associated with older age (65.2 ± 1.5 vs. 59.3 ± 1.5, p = 0.007); type 2 diabetes mellitus (33.8% vs. 17.9%; p = 0.019); hypertension (63.2% vs. 39.3%; p = 0.003); metabolic syndrome (85.3% vs. 64.3%; p = 0.003); and higher VFA/SFA ratio (1.0 ± 0.1 vs. 0.7 ± 0.1; p < 0.001). In multivariate analysis, metabolic syndrome (odds ratio [OR]: 2.5; 95% confidence interval [CI]: 1.1-5.7; p = 0.035) and higher VFA/SFA ratio (OR: 3.3; 95% CI: 1.6-7.2; p = 0.002) remained independently associated with SIBO. The presence of SIBO was found to be associated with high VFA/SFA ratio measured from cross-sectional CT image.


Best practices in assessing cardiac protein O-GlcNAcylation by immunoblot.
The modification of serine and threonine amino acids of proteins by O-linked N-acetylglucosamine (O-GlcNAc) regulates the activity, stability, function, and subcellular localization of proteins. Dysregulation of O-GlcNAc homeostasis is well established as a hallmark of various cardiac diseases, including cardiac hypertrophy, heart failure, complications associated with diabetes, and responses to acute injuries such as oxidative stress and ischemia-reperfusion. Given the limited availability of site-specific O-GlcNAc antibodies, studies of changes in O-GlcNAcylation in the heart frequently use pan-O-GlcNAc antibodies for semiquantitative evaluation of overall O-GlcNAc levels. However, there is a high degree of variability in many published cardiac O-GlcNAc blots. For example, many blots often have regions that lack O-GlcNAc positive staining of proteins either below 50 or above 100 kDa. In some O-GlcNAc blots, only a few protein bands are detected, while in others, intense bands around 75 kDa dominate the gel due to nonspecific IgM band staining, making it difficult to visualize less intense bands. Therefore, the goal of this study was to develop a modifiable protocol that optimizes O-GlcNAc positive banding of proteins in cardiac tissue extracts. We showed that O-GlcNAc blots using CTD110.6 antibody of proteins ranging from <30 to ∼450 kDa could be obtained while also limiting nonspecific staining. We also show that some myofilament proteins are recognized by the CTD110.6 antibody. Therefore, by protocol optimization using the widely available CTD110.6 antibody, we found that it is possible to obtain pan-O-GlcNAc blots of cardiac tissue, which minimizes common limitations associated with this technique.NEW & NOTEWORTHY The post-translational modification of proteins by O-linked N-acetylglucosamine (O-GlcNAc) is recognized as mediating cardiac pathophysiology. However, there is considerable variability in the quality of O-GlcNAc immunoblots used to evaluate changes in cardiac O-GlcNAc levels. Here we show that with relatively minor changes to a commonly used protocol it is possible to minimize the intensity of nonspecific bands while also reproducibly generating O-GlcNAc immunoblots covering a range of molecular weights from <30 to ∼450 kDa.


[A clinical study of flap necrosis protection with dexamethasone].
This clinical study was to find an effective method to prevent flap from necrosis. When a flap showed blood flow problems, dexamethasone of 0.3 mg/kg was given intravenously for 2 days. Then the dose was decreased to 0.2 mg/kg at the third and the fourth day. At the fifth and the sixth day, dexamethasone of 0.1 mg/kg was given. A total of 51 flaps with blood flow problems was treated with this method. Thirty of them recovered completely; 18 survived mostly and 3 failed. Early application of dexamethasone can effectively ameliorate blood flow problems of the flap.


Functional profile of CD4+ and CD8+ T cells in latently infected individuals and patients with active TB.
Tuberculosis (TB) is one of the most important infectious diseases around the world. Several studies have focused on the identification of correlates of protection against TB. Most of them have concentrated on the study of IFN-γ due to its robust association with protection against TB. However, given the complexity of the immune response elicited after Mtb infection, other cytokines should also be considered. In the present study, we evaluated Th1 and Th17 responses and their association with the protection or development of active disease. Therefore, non infected individuals (nonTBi), latently infected individuals (LTBi) and patients with active TB (ATB) were studied. The evaluation of the number of cytokine producing cells by ELISPOT showed a higher number of IFN-γ-producing cells in ATB patients, but no differences were found regarding the number of IL-17 producing cells among studied groups. The evaluation of IFN-γ, IL-2, TNF-α and IL-17 producing CD4+ and CD8+ T cells at 1 day and 6 days of stimulation with mycobacterial antigens suggests the presence of functional signatures associated with latency or active TB. The results presented herein suggest the possible use of the evaluation of Th1-type cytokines, such as IFN-γ and/or TNF-α, as a correlate of protection against TB; however, these results need to be validated for other groups.


Acaricidal activity of extract from Eupatorium adenophorum against the Psoroptes cuniculi and Sarcoptes scabiei in vitro.
The possible acaricidal activity of Eupatorium adenophorum was analyzed using extracts created by water decocting, ethanol thermal circumfluence, and steam distillation. The toxic effect of each extract was tested against Psoroptes cuniculi and Sarcoptes scabiei in vitro. Ethanol thermal circumfluence extract had strong toxicity against mites, killing all S. scabiei at 0.5 and 1.0 g/ml (w/v) concentration, while 1g/ml extract was also found to kill all P. cuniculi within a 4-h period. Similarly, 0.25, 0.5 and 1.0 g/ml concentration of extract had strong toxicity against S. scabiei, with median lethal time (LT(50)) values at 0.866, 0.785 and 0.517 h, respectively. 0.5 g/ml and 1g/ml showed strong acaricidal action against P. cuniculi; the LT(50) values were 0.93 h and 1.29 h, respectively. The median lethal concentration (LC(50)) values were 0.22 g/ml for Scabies mite and 0.64 g/ml for P. cuniculi in 1h. The results indicated that E. adenophorum contains potent acaricidal ingredients; as a first step in the potential development of novel drugs, it may provide new acaricidal compounds for the effective control of animal acariasis.


5,6-Dimethyl-1,2,4-triazin-3-amine.
In the crystal structure of the title compound, C(5)H(8)N(4), adjacent mol-ecules are connected through N-H⋯N hydrogen bonds, resulting in a zigzag chain along [100]. The amino groups and heterocyclic N atoms are involved in further N-H⋯N hydrogen bonds, forming R(2) (2)(8) motifs.


The Association Between Socioeconomic Factors at Diagnosis and Survival in Medulloblastoma: A Propensity Score-Matched Analysis and Population-Based Study.
Medulloblastoma (MB) is the most common malignant brain tumor of childhood. The associations between socioeconomic statuses (SES) and survival outcomes of medulloblastoma remain unclear. The aim of this study was to develop a nomogram to predict medulloblastoma specific death (MBSD) and overall survival (OS) in patients with medulloblastoma, taking into account socioeconomic factors in patients with medulloblastoma. We included patients diagnosed with MB between 1975 and 2016 from the Surveillance, Epidemiology, and End Results database. Propensity Score Matching (PSM) was performed to reduce selection bias. Multivariate cox proportional hazards model was used to assess SES impact and clinically relevant variables of medulloblastoma specific death and overall survival. Independent prognostic factors determined by multivariate analysis were used to construct nomograms. A total of 2660 patients were enrolled after matching. Study showed unemployed rate (MBSD, high level vs. low level, P = 0.020) (OS, high level vs. low level, P = 0.017), and marital status (OS, married vs unmarried/unknown, P = 0.029) were important factors affecting prognosis of medulloblastoma in male. Meanwhile, median household income (MBSD, quartile 1 vs. quartile 3, P = 0.047) (OS, quartile 1 vs. quartile 2, P = 0.017) (OS, quartile 1 vs. quartile 3, P = 0.014), residence (MBSD, urban vs. rural, P = 0.041), and insurance status (MBSD, insured vs. uninsured/unknown, P = 0.002)(OS, insured vs. uninsured/unknown, P = 0.001) were significant factors affecting prognosis of medulloblastoma in female. Through the calibration plot and C-index test, our nomogram was also of predictive significance. The unique features of MB have provided a scenario for analysis of the impact of racial, ethnic, gender, and socioeconomic factors. The current findings have important public health implications for achieving the goal of a healthy population. Given the known morbidity rates, long-term psychological, financial and medical burdens that these children and their families must bear, it is critical to identify and address these gaps.


Determination of 2,4-diamino-5-(3,4-dichlorophenyl)-6-methylpyrimidine (BW 197U) in human plasma.
A gas-liquid chromatographic method for the measurement of 2,4-diamino-5-(3,4-dichlorophenyl)-6-methylpyrimidine (BW 197U) in human plasma has been developed. After extraction from alkaline medium into ethyl acetate the compound is injected into a gas-liquid chromatograph and measured using a 63Ni constant-current electron-capture detector. The range of concentrations measured was from 10 ng/ml to 10 microgram/ml in plasma. An internal standard was employed and reproducibility between replicates was found to be good. The method has been semi-automated by the use of an auto-sampler controlled by a minicomputer which also processes the data.


Supporting breastfeeding mothers: qualitative synthesis.
This paper is a report of a synthesis of mothers' and healthcare professionals' experiences and perceptions of breastfeeding support. Despite increasing knowledge, breastfeeding rates remain relatively static and mothers continue to report dissatisfaction with their experiences of breastfeeding. Greater understanding of breastfeeding may be achieved through rigorous qualitative research, and there has been a recent increase in such studies. Electronic databases and citation lists of published papers were searched for articles listed between 1990 and 2005 and updated in May 2007. Studies were included if they used qualitative methods, were published in English, explored an aspect of breastfeeding and were based in a westernized country. Papers were included if they reported studies using qualitative methods to explore breastfeeding and were published in English and based in a westernized country. Each study was reviewed and assessed independently, key themes extracted and grouped, and secondary thematic analysis used to explore key concepts. From the 1990-2005 search, five themes emerged in health service support of breastfeeding: the mother-health professional relationship, skilled help, pressures of time, medicalization of breastfeeding and the ward as a public place. Social support had two themes: compatible and incompatible support. One additional theme emerged from the update to 2007: health professional relationships. Mothers tended to rate social support as more important than health service support. Health service support was described unfavourably with emphasis on time pressures, lack of availability of healthcare professionals or guidance, promotion of unhelpful practices and conflicting advice. Changes are required within the health services to address the needs of both mothers and staff.


The identification of CD163 expressing phagocytic chondrocytes in joint cartilage and its novel scavenger role in cartilage degradation.
Cartilage degradation is a typical characteristic of arthritis. This study examined whether there was a subset of phagocytic chondrocytes that expressed the specific macrophage marker, CD163, and investigated their role in cartilage degradation. Cartilage from the knee and temporomandibular joints of Sprague-Dawley rats was harvested. Cartilage degradation was experimentally-induced in rat temporomandibular joints, using published biomechanical dental methods. The expression levels of CD163 and inflammatory factors within cartilage, and the ability of CD163(+) chondrocytes to conduct phagocytosis were investigated. Cartilage from the knees of patients with osteoarthritis and normal cartilage from knee amputations was also investigated. In the experimentally-induced degrading cartilage from temporomandibular joints, phagocytes were capable of engulfing neighboring apoptotic and necrotic cells, and the levels of CD163, TNF-α and MMPs were all increased (P<0.05). However, the levels of ACP-1, NO and ROS, which relate to cellular digestion capability were unchanged (P>0.05). CD163(+) chondrocytes were found in the cartilage mid-zone of temporomandibular joints and knee from healthy, three-week old rats. Furthermore, an increased number of CD163(+) chondrocytes with enhanced phagocytic activity were present in Col-II(+) chondrocytes isolated from the degraded cartilage of temporomandibular joints in the eight-week experimental group compared with their age-matched controls. Increased number with enhanced phagocytic activity of CD163(+) chondrocytes were also found in isolated Col-II(+) chondrocytes stimulated with TNF-α (P<0.05). Mid-zone distribution of CD163(+) cells accompanied with increased expression of CD163 and TNF-α were further confirmed in the isolated Col-II(+) chondrocytes from the knee cartilage of human patients with osteoarthritis, in contrast to the controls (both P<0.05). An increased number of CD163(+) chondrocytes with enhanced phagocytic activity were discovered within degraded joint cartilage, indicating a role in eliminating degraded tissues. Targeting these cells provides a new strategy for the treatment of arthritis.


High circulating adiponectin levels predict decreased muscle strength among older adults aged 70 years and over: A prospective cohort study.
Population-based researches indicate that circulating adiponectin is inversely associated with muscle strength. However, interpretation of the findings has been limited by the use of a cross-sectional design. This study aimed to examine the prospective relationship between baseline circulating adiponectin concentration and change in muscular function-related physical performance in older adults. A 1-year prospective cohort study of Japanese community-dwelling elderly was conducted between 2002 and 2003. Four hundred thirty-four older persons participated in the measurements of physical function, including leg extension power, functional reach, timed up-and-go test, and 10-m maximum walking speed, at baseline and follow-up. After adjustment for potential covariates, higher serum adiponectin concentration was found to be significantly associated with poorer physical performance at baseline (leg extension power [watt], P < 0.001; functional reach [cm], P < 0.001; log timed up-and-go test, P = 0.007; log 10-m maximum walking speed, P < 0.001). The results of the prospective analysis by analysis of covariance indicated that the elderly with higher serum adiponectin concentrations (tertiles) at baseline tended to have a decreased performance in leg extension power (means [95% confidence interval]: lowest, -105 [-125, -85.7]; middle, -117 [-135, -97.8]; highest, -140 [-160, -120], watt, P for trend = 0.021) and timed up-and-go test (lowest, -0.08 [-0.28, -0.12]; middle, -0.10 [-0.29, 0.10]; highest, 0.28 [0.07, 0.48], s, P for trend = 0.019), but not two other functioning. High circulating adiponectin concentration may be an indicator of decreased physical performance, especially muscle strength, in older adults.


Controversies in the pathophysiology and management of hyphema.
Traumatic hyphemas present dilemmas to physicians. There are numerous controversies pertaining to the optimal approach to traumatic hyphema and no standardized guidelines for its management. We address some of these controversies and present a pragmatic approach. We discuss various medical agents and surgical techniques available for treatment, along with the indications for their use. We address the complications associated with hyphema and how to diagnose and manage them and consider the management of hyphema in special situations such as in children and sickle-cell anemia and in rare clinical syndromes such as recurrent hyphema after placement of anterior chamber intraocular lenses.


'Hospitals of excellence' outshine others in mortality, complications.
The latest Hospital Quality and Clinical Excellence study from HealthGrades, Inc. finds that 152,966 lives could have been saved and 21,896 complications avoided. Leadership by upper management remains a critical element for quality improvement. Leading hospitals regularly use multidiscipline approach and implement new strategies such as rapid response teams.


GABAB Receptors and Alcohol Use Disorders: Preclinical Studies.
Preclinical research over the past several decades has demonstrated a role for the γ-aminobutyric acidB (GABAB) receptor in alcohol use disorder (AUD). This chapter offers an examination of preclinical evidence on the role of the GABAB receptor on alcohol-related behaviors with a particular focus on the GABAB receptor agonist baclofen, for which effects have been most extensively studied, and positive allosteric modulators (PAMs) of the GABAB receptor. Studies employing rodent and non-human primate models have shown that activation of the GABAB receptor can reduce (1) stimulating and rewarding effects of alcohol; (2) signs of alcohol withdrawal in rats made physically dependent on alcohol; (3) acquisition and maintenance of alcohol drinking under a two-bottle alcohol versus water choice procedure; (4) alcohol intake under oral operant self-administration procedures; (5) motivational properties of alcohol measured using extinction and progressive ratio procedures; (6) the increase in alcohol intake after a period of alcohol abstinence (the alcohol deprivation effect or ADE); and (7) the ability of alcohol cues and stress to reinstate alcohol seeking when alcohol is no longer available. Baclofen and GABAB PAMs reduce the abovementioned behaviors across different preclinical models, which provides strong evidence for a significant role of the GABAB receptor in alcohol-related behaviors and supports development of medications targeting GABAB receptors for the treatment of AUD. This chapter highlights the value of examining mechanisms of alcohol-related behaviors across multiple animal models to increase the confidence in identification of new therapeutic targets.


Microflora profiling of infected root canal before and after treatment using culture-independent methods.
This study aimed to profile the microflora in infected root canals before and after root canal treatment using culture-independent methods. Six infected root canals in single-rooted teeth with periapical lesions from five subjects were included. Quantification of total bacteria was performed by real-time PCR with primers targeting 16S rRNA genes. PCR products with universal 16S rRNA gene primers were cloned and partially sequenced, and bacterial identification at the species level was performed by comparative analysis with the GenBank database. The concentration of extracted DNA before treatment was higher than that after root canal treatment, although the difference was not statistically significant. Sequence analysis revealed that oral bacteria such as Fusobacterium, Streptococcus, Olsenella, and Pseudoramibacter detected in cases before root canal treatment disappeared after treatment. These results suggest that the root canal microflora are distinct before and after root canal treatment, and that treatment changes the microflora in both quantity and quality.


[Kaufman Assessment Battery for Children. Differential evaluation of (partial) intellectual ability of mentally handicapped adults].
The use of the Kaufman-Assessment Battery for Children (K-ABC) was evaluated with a sample of 50 mentally disabled adults in a wide age range, as there exist no special tests for this group of patients. Intercorrelation--and factor analyses, and correlations with the German adaptation of the Wechsler Adult Intelligence Scale (HAWIE-R) proved the use of the K-ABC to be valid. As an assessment battery for children designed for the wide age range from 2-6 to 12-5 years is more adequate to the capability of intellectually impaired adults, the K-ABC delivers results on individual strengths and weaknesses, based on a neuropsychological theory. Thus the individual cognitive abilities can be assessed with more differentiation, and that gives an important base for intervention, partial remediation, and placement.


Contribution of cathepsin B-dependent Nlrp3 inflammasome activation to nicotine-induced endothelial barrier dysfunction.
Recent studies indicate that endothelial Nlrp3 inflammasome is critically involved in the development of cardiovascular complications. However, it remains unknown whether endothelial inflammasome is involved in endothelial barrier dysfunction associated with smoking. This study aims to investigate the role of endothelial Nlrp3 inflammasome in nicotine-induced disruption of inter-endothelial tight junctions and consequent endothelial barrier dysfunction. The confocal microscopic analysis demonstrated that mice treated with nicotine exhibited disrupted inter-endothelial tight junctions as shown by decreased ZO-1 and ZO-2 expression in the coronary arterial endothelium, whereas the decreases in ZO-1/2 were prevented by Nlrp3 gene deficiency. In cultured endothelial cells, nicotine caused Nlrp3 inflammasome complex formation and enhances the inflammasome activity as shown by increased cleavage of pro-caspase-1, and interleukin-1β (IL-1β) production. Further, nicotine disrupted tight junction and increased permeability in an endothelial cell monolayer, and this nicotine-induced effect was prevented by silencing of Nlrp3 gene, inhibition of caspase-1, or blockade of high mobility group box 1 (HMGB1). Nicotine increased endothelial cell lysosomal membrane permeability and triggered the lysosomal release of cathepsin B, whereas these events were prevented by pretreating cells with a lysosome stabilizing agent, dexamethasone. Collectively, our data suggest that nicotine enhances cathepsin B-dependent Nlrp3 inflammasome activation and the consequent production of a novel permeability factor HMGB1, which causes disruption of inter-endothelial tight junctions leading to endothelial hyperpermeability. Instigation of endothelial inflammasomes may serve as an important pathogenic mechanism contributing to the early onset of vasculopathy associated with smoking.


Helicobacter pylori activates NF-κB by inducing Ubc13-mediated ubiquitination of lysine 158 of TAK1.
The Helicobacter pylori virulence factor CagA targets a variety of host proteins to alter different cellular responses, including the induction of pro-inflammatory cytokines. We have previously shown that CagA-facilitated lysine 63-linked ubiquitination of TAK1 is essential for the H. pylori-induced NF-κB activation and the expression of proinflammatory cytokines. However, the molecular mechanism for TAK1 ubiquitination and activation in H. pylori-mediated NF-κB activation remains elusive. Here, we identify lysine 158 of TAK1 as the key residue undergoing lysine 63-linked ubiquitination in response to H. pylori infection. Mutation of lysine 158 to arginine prevents the ubiquitination of TAK1 and impairs H. pylori-induced TAK1 and NF-κB activation. Moreover, we demonstrate that E2 ubiquitin conjugating enzyme Ubc13 is involved in H. pylori-mediated TAK1 ubiquitination. Suppressing the activity of Ubc13 by a dominant-negative mutant or siRNA abolishes CagA-facilitated and H. pylori-induced TAK1 and NF-κB activation. These findings further underscore the importance of lysine 63-linked ubiquitination of TAK1 in H. pylori-induced NF-κB activation and NF-κB-mediated inflammatory response.


[Fanconi's anemia: in vitro tests for the individualization of heterozygotes].
Fanconi's anaemia (FA) is an autosomal recessive mutation associated with constitutional chromosome aberrations. Patient's cells show increased susceptibility to different mutagens, especially bifunctional alkylating agents. Asymptomatic heterozygotes have a population prevalence of 1 in 300, and have been considered at risk for cancers. It has also been shown that their cells are especially sensitive to some chemicals in vitro. Laboratory tests are presented and discussed which are adequate for FA heterozygotes identification and for understandings the basic defect of this mutation.


High genetic risk individuals benefit less from resistance exercise intervention.
Genetic factors have an important role in body mass index (BMI) variation, and also likely have a role in the weight loss and body composition response to physical activity/exercise. With the recent identification of BMI-associated genetic variants, it is possible to investigate the interaction of these genetic factors with exercise on body composition outcomes. In a block-randomized clinical trial of resistance exercise among women (n=148), we examined whether the putative effect of exercise on weight and dual-energy x-ray absorptiometry-derived body composition measurements differs according to genetic risk for obesity. Approximately one-half of the sample was randomized to an intervention consisting of a supervised, intensive, resistance exercise program, lasting 1 year. Genetic risk for obesity was defined as a genetic risk score (GRS) comprised of 21 single-nucleotide polymorphisms (SNPs) known to be associated with BMI variation. We examined the interaction of exercise intervention and the GRS on anthropometric and body composition measurements after 1 year of the exercise intervention. We found statistically significant interactions for body weight (P=0.01), body fat (P=0.01), body fat % (P=0.02) and abdominal fat (P=0.02), whereby the putative effect of exercise is greater among those with a lower level of genetic risk for obesity. No single SNP appears to be a major driver of these interactions. The weight-loss response to resistance exercise, including changes in body composition, differs according to an individual's genetic risk for obesity.


Comparative evaluation of the effect of cyclodextrins and pH on aqueous solubility of apigenin.
The aqueous solubility of a flavonoid, apigenin, was studied in the presence of first generation cyclodextrins (α-CyD, β-CyD, γ-CyD), ionic and nonionic synthetic derivatives of β-CyD, namely SBE-β-CyD, HP-β-CyD and RM-β-CyD at various physiological pH. The order of solubility enhancement was as follows: RM-β-CyD>SBE-β-CyD>γ-CyD>HP-β-CyD>β-CyD>α-CyD. The phase solubility diagrams of HP-β-CyD and SBE-β-CyD indicated Higuchi AL subtype behavior, suggesting 1:1 stoichiometry of the complex. In contrast, AP subtype, so higher order complex formation can be assumed in the case of RM-β-CyD and γ-CyD. The formation of inclusion complexes has been confirmed by absorption and fluorescence spectroscopic measurements. Increased antioxidant activity was observed due to the inclusion complexes. These results prove that synthetic derivatives of β-CyD will be potentially useful excipients in the development of drug delivery systems for healthcare products containing flavonoids.


Strains ofaspergillus versicolor tiraboschi synthesizing sterigmatocystin and the differentiation of mycotoxic risk dependent on their productivity in housing buildings.
In biomasses from 22 moulds isolated in housing buildings a Chromatographie HPLC UV/VIS analysis was carried out to assess the production of sterigmatocystin by fungal isolates. The results are discussed with respect to mycotoxic risk for people exposed to the presence ofAspergillus versicolor. This fungus is often present on building materials, but does not always produce the carcinogenic mycotoxin sterigmatocystin (ST). In this study, 19 (86%) of the 22 strains synthesized ST at detectable levels (>0.03 mg kg-1 biomass). Three of the strains (14%) were found to synthesize ST at levels exceeding 100 mg kg-1 of the air-dry mould biomass with laboratory medium. One strain proved to be highly productive and synthesized more than 500 mg kg-1 ST. Thus, it presented the highest mycotoxic danger for the dwellers of the buildings whereA. versicolor infested the walls. However, most strains are low producers of ST, with 12 of 22 isolates synthesizing less than 10 mg kg-1 biomass. Mycotoxigenic and highly productive strains that produced significant amounts of ST (>500 mg kg-1 biomass) were less frequently found, with approximately 5% of all isolates from buildings studied for ST production.


Examination of the Flynn effect in German patients with epilepsy assessed with the Wechsler Adult Intelligence Scale (WAIS) III and IV.
The Flynn effect describes an increase in intelligence quotient (IQ) in the general population of about 3 points per decade. While this effect is well established in healthy individuals, research exploring the link to brain pathologies is scarce. We investigated the Flynn effect in a German sample of 203 patients with epilepsy with left, right, and bilateral lesions. Intelligence quotient values were obtained using the Wechsler Adult Intelligence Scales (WAIS) III and IV. Our results showed a stable Flynn effect with nearly no difference in adjusted full scale IQ (FSIQ) scores (0.02 IQ points) between the WAIS-III and WAIS-IV samples. There were no significant interactions between the side of pathology and corrected IQ values. Our sample showed a tendency towards performing worse in the WAIS-IV in three out of four subscales independently of the Flynn effect, pointing out methodological differences between the newer Wechsler editions. However, although patients with bilateral lesions performed worst across all subscales, they exhibited a similar pattern as patients with lesions in the left or right hemisphere, indicating that also more severe forms of brain pathologies can profit from the mechanisms behind the Flynn effect.


The vascular supply to the spinal cord and its relationship to anterior spine surgical approaches.
The understanding of vascular supply to the spinal cord is important given that the evolution of surgical approaches to the spine may bring along the potential for more frequent complications, especially a rare but devastating complication: that of spinal cord ischemia or infarction. To maximally avoid this complication, the relationship between the spinal cord vascularity and the anterior spine surgical approach needs further study. To provide a theoretical basis that will allow the spinal surgeon to take appropriate steps to avoid spinal cord ischemia during anterior spinal surgery through anatomic means. Spinal cord vascular casting assessment with cadaveric specimen. Twenty adult cadaveric specimens (11 men and 9 women) were obtained for the latex perfusion and vessel dissection. In addition, nine patients (seven men and two women) underwent superselective angiography of the spinal cord. The segmental arterial anastomosis and radiculomedullary vessels in the thoracolumbar region were shown and reviewed. There were approximately 21 pairs of segmental arteries in the thoracolumbar region. Adjacent segmental arteries were networked with each other. The latex infusion specimens demonstrated 72 anterior radiculomedullary arteries and 177 posterior radiculomedullary arteries in all 20 samples. The anterior and posterior spinal arteries were also networked with each other at several levels. Superselective spinal angiography was consistent with the latex infusion specimens showing. The variety of anatomy of spinal cord arterial networks is shown, and the relation between the blood supply of certain spinal levels and the potential ischemic complications during the anterior surgical approach is discussed. It is hopefully of benefit to surgeons, after fully understanding the anatomy of these spinal vascular supply structures, that there may be even greater avoidance of vascular compromise in these challenging operations.


Diagnostic value of platelet indices in COVID 19 infection: a case-control study from a single tertiary care center.
Platelets are not only involved in hemostasis and coagulation, but play a significant role in innate immunity and inflammatory response. Excess production of cytokines and acute phase reactants affect megakaryopoiesis resulting in the release of immature platelets from the bone marrow altering platelet indices. To study platelet indices in RT-PCR-proven COVID patients and non-COVID patients. A case-control study was conducted on 199 COVID-19 patients and 198 normal individuals. Blood samples were analyzed in an automated hematology analyzer. The platelet indices like platelet count, mean platelet volume (MPV), platelet distribution width (PDW), platelet large cell count (PLCC), and platelet large cell ratio (P-LCR) were compared among two groups. Platelet count in COVID-19 patients were significantly low (p<0.01) compared to controls, and a significant number of COVID-19 patients had thrombocytopenia. Plateletcrit (PCT) was also significantly decreased in COVID-19 patients compared to non-COVID individuals. MPV, PDW, and PLCR were significantly (p<0.05) high in COVID-19 patients in comparison to controls, but was not significantly raised in a large number of cases. In contrast, there were no significant differences in platelet large cell count (PLCC) values between COVID-19 cases and non-COVID-19 controls. Platelet indices like platelet count, PCT, MPV, PDW, and P-LCR are significantly altered in COVID-19 infection and thereby can be used as biomarkers in COVID-19. Further research is needed to find if these simple, cost-effective parameters can be used to predict the severity and prognosis in COVID-19 infection.


Alpha-tocopherol in the brain tissue preservation of stroke-prone spontaneously hypertensive rats.
Oxidative stress has an important role in neuronal damage during cerebral ischemia and can lead to cognitive and behavioral impairment. Alpha-tocopherol, a powerful antioxidant, may be able to preserve neuronal tissue and circumvent neurological deficits. Thus, this study aimed to investigate the influence of alpha-tocopherol in the preservation of brain tissue and the maintenance of memory formation in stroke-prone spontaneously hypertensive rats (SHRSP). To achieve this aim, twenty-four 15-week-old male SHRSP rats were separated into the following four groups (n = 6 each) that received different treatments over a 4-week period: the alpha-tocopherol group, the control group, the L-NAME group, and the L-NAME + alpha-tocopherol group. We evaluated the physiological parameters (body weight, diuresis, and food and water intake), an oxidative stress marker (malondialdehyde levels), and neurological responses (the Morris Water Maze and Novel Objects Recognition tests). Afterwards, the brains were removed for histopathological analysis and quantification of the number of cells in the hippocampus. Statistically, the alpha-tocopherol group demonstrated better results when compared to all groups. The data indicated a reduction in oxidative stress and the preservation of neurological responses in groups treated with alpha-tocopherol. In contrast, the L-NAME group exhibited increased malondialdehyde levels, impairment of neurological responses, and several hippocampus tissue injuries. The others groups exhibited nerve tissue changes that were restricted to the glial nodes. No significant alterations were observed in the physiologic parameters. Based on these findings, we suggest that alpha-tocopherol can prevent stroke, preserve the structure of the hippocampus, and maintain both memory and cognition functions.


Nonlinear relationship between level of blood flow and skin temperature for different dynamics of temperature change.
We present a study of the relationship between blood flow and skin temperature under different dynamics of skin-temperature-change: locally induced thermal shock and well controlled, gradual change. First, we demonstrate memory phenomena for blood flow and skin temperature under both conditions. Secondly, we point out that the "hysteresis" loops obtained are dependent on initial conditions, indicating physiological response times of more than twenty minutes. We also show that under thermal shock the level of blood flow is preserved up to some characteristic temperature limit, independently of subject.


Toll-like receptor 4 expression on circulating leucocytes in hemolytic uremic syndrome.
Lipopolysaccharide stimulation of toll-like receptor 4 (TLR4) activates signal transduction pathways leading to proinflammatory cytokine secretion. We investigated TLR4 surface receptor expression on peripheral blood neutrophils and monocytes and their ability to modulate inflammatory cytokine release in 15 patients 1, 3, and 10 days after hemolytic uremic syndrome (HUS) onset. Seven patients with Escherichia coli (EHEC)-associated diarrhea and seven healthy controls were also studied. Isolated leucocytes from HUS-onset patients exhibited significantly higher messenger RNA (mRNA) TLR4 expression than controls. Moreover, TLR4 protein expression on neutrophils, determined by flow cytometry, was upregulated, driving dependent proinflammatory cytokine, tumor necrosis factor alpha (TNF-α), and interleukin 8 (IL-8) increase, and decreased anti-inflammatory IL-10 release at HUS onset compared with patients with EHEC diarrhea and controls. TLR4 expression on neutrophils was positively correlated with serum TNF-α levels. Conversely, significant reduction of neutrophil TLR4 receptor expression and lack of cytokine-responsive element activation was shown in patients 3 and 10 days after HUS onset. No differences were demonstrated in TLR4 receptor expression on monocytes among the studied groups. Our results suggest TLR4 expression may be differently regulated on neutrophils and monocytes. They could be dynamically modulated across the early development of HUS on neutrophils, resulting in negative regulation preceded by TLR4 overactivation.


Volume-regulatory taurine release from a human lung cancer cell line. Evidence for amino acid transport via a volume-activated chloride channel.
Exposure of a human lung epithelial cancer cell-line to hypo-osmotic media led to a marked increase in the rate of efflux from the cells of taurine, a non-essential sulfonic amino acid. The osmotically-activated taurine efflux was inhibited by a range of known Cl- channel blockers, the most potent of which were NPPB and 1,9-dideoxyforskolin. These reagents were similarly effective at inhibiting the osmotically-activated efflux of I-, a known substrate of volume-activated Cl- channels. The results are consistent with the hypothesis that volume-regulatory taurine release from these cells is mediated by a volume-activated Cl- channel.


The role of microbe-microplastic associations in marine Nematode feeding behaviors.
Fauna across many taxa and trophic levels have been shown to consume microplastics (MPs) in experiments, providing evidence that supports field-based gut content assessments. Multiple explanations exist regarding why fauna consume MPs, one of which posits that microbial growth on MPs may facilitate faunal ingestion. However, laboratory assessments on the reasons why MPs are consumed remain limited. Here, we assessed if the presence of microbes on MPs altered marine nematode feeding behaviors across current and potential future concentrations of MPs in a local system. We used a microcosm experiment in which field-collected sediment was spiked with bacterially treated or untreated fluorescent plastic microbeads (1.0-5.0 μm) in concentrations of 102, 104, and 106 per microcosm, representing local and potential future concentrations of MPs. Ingestion by the dominant interstitial fauna was investigated after 0, 3, and 7 days using bright field microscopy. Nematodes were the only fauna across microcosms that consumed MPs, but this consumption was variable and there were no apparent trends across exposure time, bacterial treatment, or MP concentration. There were also no genera- or feeding-type-specific trends in the number of MPs consumed, though four of the top five nematode genera that consumed MPs were pollution-tolerant genera. Our study demonstrates that microbe-MP associations do not drive marine nematodes to eat MPs, especially at local field concentrations. While there were no trends across any of the nematode genera in our study, we recognize that unrealistic MP concentrations in other studies may provide alternative explanations for nematode consumption of MPs.


Clinical evaluation of the Mannheim Prognostic Index in post-operative peritonitis: a prospective cohort study.
Postoperative peritonitis (PoP), despite their relatively low incidence, are associated with high mortality. Such poor outcomes are also related to the high proportion of aged patients, whose intra-abdominal infections are difficult to manage. The study included 84 consecutive patients with PoP. The aim was the validation of the Mannheim Prognostic Index (MPI) in the context of PoP and the assessment of the prognostic impact of age and other clinical factors in a large series from a tertiary center. PoP had an incidence of 3.9% in all the abdominal surgeries in the study period. Surgical control of POP focus was achieved in 90.5% of cases and a complete abdominal clearance in 58.3%. Complication rate was 75% with a mortality of 26.2%. For MPI score, the ROC curve indicated a cut-off value of 29 with a sensitivity of 72.7% and specificity of 67.7% in predicting death. At univariate analysis, factors significantly related to poorer prognosis included advanced age (p 0.001), site of primary surgery (p 0.05), lack of abdominal clearance (p 0.003), generalized peritonitis (p 0.04) and high MPI score (p < 0.001). Age, MPI score and absence of abdominal clearance resulted in independent prognostic factors at multivariate analysis. MPI showed good efficacy in identifying POP patients at high risk of death. The increased risk of mortality related to advanced age should be considered with MPI score in planning the treatment. An aggressive and early diagnostic-therapeutic approach is required to reduce the MPI score and improve the prognosis.


[Dysequilibrium in idiopathic Parkinson disease. The effect of cerebrovascular comorbidity].
Disturbance of balance in idiopathic Parkinson's disease (IPD) has a significant effect on disability. Yet the underlying mechanisms and the contribution of age-associated comorbidity to dysequilibrium are unclear. In this study, static posturography was performed in 30 healthy controls and 40 patients with IPD. Comparison of sway during quiet stance did not show significant differences between patients and controls. Multiple linear regression analysis was used to identify factors responsible for the considerable interindividual variance in patients. Results of the pull test, CT-verified cerebral microangiopathy, dementia, and age were assessed, but only cerebrovascular comorbidity contributed significantly to variance. Apart from increased sway, patients with coinciding cerebral microangiopathy (n = 20) more frequently had a history of falls or pathological responses in the pull test. The present results suggest that cerebrovascular comorbidity enhances dysequilibrium in IPD. Pathological sway in IPD can indicate comorbidity and may have implications for further diagnostics.


Myeloperoxidase Inhibition Reverses Biomarker Profiles Associated With Clinical Outcomes in HFpEF.
Systemic microvascular dysfunction and inflammation are postulated to play a pathophysiologic role in heart failure with preserved ejection fraction (HFpEF). This study aimed to identify biomarker profiles associated with clinical outcomes in HFpEF and investigate how inhibition of the neutrophil-derived reactive oxygen species-producing enzyme, myeloperoxidase, affects these biomarkers. Using supervised principal component analyses, the investigators assessed the associations between baseline plasma proteomic Olink biomarkers and clinical outcomes in 3 independent observational HFpEF cohorts (n = 86, n = 216, and n = 242). These profiles were then compared with the biomarker profiles discriminating patients treated with active drug vs placebo in SATELLITE (Safety and Tolerability Study of AZD4831 in Patients With Heart Failure), a double-blind randomized 3-month trial evaluating safety and tolerability of the myeloperoxidase inhibitor AZD4831 in HFpEF (n = 41). Pathophysiological pathways were inferred from the biomarker profiles by interrogation of the Ingenuity Knowledge Database. TNF-R1, TRAIL-R2, GDF15, U-PAR, and ADM were the top individual biomarkers associated with heart failure hospitalization or death, and FABP4, HGF, RARRES2, CSTB, and FGF23 were associated with lower functional capacity and poorer quality of life. AZD4831 downregulated many markers (most significantly CDCP1, PRELP, CX3CL1, LIFR, VSIG2). There was remarkable consistency among pathways associated with clinical outcomes in the observational HFpEF cohorts, the top canonical pathways being associated with tumor microenvironments, wound healing signaling, and cardiac hypertrophy signaling. These pathways were predicted to be downregulated in AZD4831 relative to placebo-treated patients. Biomarker pathways that were most strongly associated with clinical outcomes were also the ones reduced by AZD4831. These results support the further investigation of myeloperoxidase inhibition in HFpEF.


New evidence for the dimeric nature of NADH:Q oxidoreductase in bovine-heart submitochondrial particles.
The initial velocity of NADH oxidation by bovine-heart submitochondrial particles was measured at pH 8.0 after pretreatment of these particles with different amounts of the inhibitor piericidine A together with 0.035 mM NADH. The amount of piericidine A required to fully inhibit the NADH oxidation activity extrapolated to exactly 1.0 per Fe-S cluster 2 of NADH:Q oxidoreductase. When no reducing equivalents from NADH were present during the pretreatment, this ratio was 1.2. The difference is explained by assuming that NADH:Q oxidoreductase binds piericidine A more effectively in the reduced state than in the oxidized state. It was also found that after Q10-extraction and reincorporation of submitochondrial particles, the amount of piericidine A required to fully inhibit the NADH oxidation activity of the particles increased with the amount of Q10 present during reincorporation. This is explained by assuming that binding of piericidine A, to the inhibitory site of NADH:Q oxidoreductase requires Q10. When 0.035 mM NADPH instead of NADH was present during the pretreatment of submitochondrial particles with piericidine A, the amount of inhibitor per cluster 2 required to fully inhibit the initial NADH-oxidation activity extrapolated to 0.5. This result strongly suggests that NADH:Q oxidoreductase is a functional dimer.


Chemodivergent Synthesis of Aza-Heterocycles with a Quarternary Carbon Center via [4 + 1] Annulation between Azoalkenes and α-Bromo Carbonyl Compounds.
An efficient [4 + 1] annulation reaction between in situ generated azoalkene intermediates and α-bromocarbonyls has been established. A series of skeletally diverse aza-heterocycles with a functionalized quaternary center were obtained in up to 89% yield under mild conditions.


Dysfunctional glucocorticoid receptor with a single point mutation ablates the CCAAT/enhancer binding protein-dependent growth suppression response in a steroid-resistant rat hepatoma cell variant.
We used glucocorticoid-resistant and -sensitive hepatoma cell variants to characterize the mechanism of hepatoma cell resistance to the growth inhibitory effects of glucocorticoids. BDS1 hepatoma cells express transcriptionally active glucocorticoid receptors and undergo a stringent G1 cell cycle arrest in response to glucocorticoids that is dependent on the induced expression of the CCAAT/enhancer binding protein alpha (C/EBPalpha) transcription factor. In contrast, EDR1 hepatoma cells, which express normal levels of glucocorticoid receptors, fail to growth arrest or express C/EBPalpha when treated with glucocorticoids. Ectopic expression of wild-type rat glucocorticoid receptors into EDR1 cells restored the growth suppression response, suggesting a defect in the EDR1 receptor. DNA sequence analysis revealed a single point mutation causing a cysteine-to-tyrosine substitution at amino acid position 457 (C457Y-GR) in the zinc finger region of the glucocorticoid receptor that mediates both receptor-DNA and receptor-protein interactions. Glucocorticoid activation of the alpha1-acid glycoprotein (AGP) promoter, a liver acute-phase response gene, requires receptor-DNA binding as well as an interaction with C/EBPalpha. In contrast to the wild-type glucocorticoid receptor, ectopic expression of C/EBPalpha in EDR1 cells, or coexpression of C/EBPalpha along with the C457Y-GR into receptor-deficient EDR3 cells was required to partially restore glucocorticoid responsiveness of the AGP promoter by the EDR1 glucocorticoid receptor. Constitutive expression of the wild-type glucocorticoid receptor, but not the C457Y-GR mutant, was sufficient to restore the glucocorticoid growth suppression response to receptor-deficient EDR3 cells. Thus, we have identified a glucocorticoid-resistant hepatoma cell variant with a single point mutation in the zinc finger region of the glucocorticoid receptor gene that ablates the glucocorticoid growth suppression response and attenuates transcriptional activation of the AGP promoter.


Regulation of mental states and biofeedback techniques: effects on breathing pattern.
The purpose of the present study was to examine whether breathing pattern may be used as a reliable index for the effectiveness of techniques applied for the regulation of mental states. Heart rate (HR), breathing pattern, galvanic skin response (GSR), and electromyogram (EMG) of the frontalis muscle were measured in 39 male and female subjects aged 18-25 years during 10-minute treatment with relaxation technique (autogenic training and/or music) followed by 10 minutes of imagery training. In the first 7 sessions biofeedback (BFB) was not included, while during the last 6 sessions BFB was introduced and utilized by the subjects. Relaxation (music or autogenic training) led to a decrease in breathing frequency, attributed to lengthening of expiration time, as well as reduced HR, GSR, and frontalis EMG response. In most instances imagery training was related to an increase in these indices. Specifically, significant tachypnea was observed during imagery of sprint running. In most cases BFB substantially augmented the physiological responses. In conclusion, our data suggest that, compared with HR, GSR, and EMG responses, the breathing pattern is at least as sensitive to the mental techniques employed, and may be useful as a psychophysiological index for diagnosis and testing, especially in sport practice.


Major depressive disorders in outpatients with cancers at the Nigerian National Hospital.
Frequently, little attention is paid by clinicians on the psychiatric disorders that may be associated with cancers. The aim of this study was to determine the prevalence of major depressive disorders in outpatients with cancers at the Radiology Oncology Centre of the National Hospital, Abuja, Nigeria. It also sought to determine the socio-demographic and clinical factors associated with the disorder. A sample of 177 randomly selected outpatients with cancers was interviewed with socio-demographic questionnaire and the major depressive episode section of the Mini International NeuroPsychiatric Interview at the hospital. The mean age (SD) of the participants was 48.9 (13.8) years, with females accounting for 61.6% of them. The prevalence of current major depressive disorder, past depressive episode and recurrent depressive disorder were 25.1%, 24.3% and 7.5%, respectively. The current depressive disorder was found to be significantly associated with the presence of disturbing pain and the stage of the disease. The occurrence of depressive disorders in patients with cancers at the hospital was high. Their assessment and treatment should be prioritized to ensure complete management of the patients with cancer conditions.


Templated synthesis of zirconium(IV)-based metal-organic layers (MOLs) with accessible chelating sites.
Metal-organic layers (MOLs) are of great interest in heterogeneous catalysis, particularly materials that can accommodate extraneous metal centres. Here, we demonstrate a two-step preorganisation/delamination synthetic strategy using CuI as a template to prepare Zr-based MOLs with accessible 'syn' bis-pyrazolyl chelating sites (named UAM-2·ns) that are poised for quantitative post-synthetic metalation with late transition metals.


Tight-junction tightness of Necturus gall bladder epithelium is not regulated by cAMP or intracellular Ca2+. I. Microscopic and general electrophysiological observations.
Following the publications by Duffey et al. [Nature 294:451 (1981)] and Palant et al. [Am J Physiol 245: C203 (1983)] it is generally accepted that tight-junction tightness of Necturus gall bladder epithelium is up-regulated by cAMP-mediated and Ca(2+)-mediated stimulation. This conclusion was mainly based on observed increases in transepithelial resistance (Rt). However, since in leaky epithelia Rt cannot be simply equated with the tight junction resistance (Rj), but may include large contributions from the lateral space resistance (Rlis), we asked whether the observed increases in Rt resulted indeed from Rj or whether Rlis also increased. The experiments were performed on Necturus gall bladders using forskolin or the Ca2+ ionophore A23187 as stimulants. Forskolin (2 mumol/l) had a biphasic effect. In the first 5 min Rt decreased from 128 +/- 13 to 119 +/- 14 omega cm2 (P < 0.05, n = 10) which probably reflects stimulation of an apical cell membrane Cl- conductance (see accompanying paper). Subsequently Rt increased in approximately 30 min to 184 +/- 20 omega cm2 and then remained fairly constant. Simultaneously the lateral spaces collapsed. If the spaces were now transiently opened by passing mucosa-positive direct current across the epithelium, Rt fell transiently to 111 +/- 7 omega cm2, but returned gradually to its elevated level when the spaces collapsed again. When the spaces were constantly dilated by a serosa-positive hydrostatic pressure of 1 cm H2O, forskolin neither affected the space width nor increased Rt, and current passage was virtually ineffective, although the cells depolarized in response to forskolin as usual.(ABSTRACT TRUNCATED AT 250 WORDS)


Crack modes and toughening mechanism of a bioinspired helicoidal recursive composite with nonlinear recursive rotation angle-based layups.
The rotation angle is an important parameter affecting the performance of helical structures, and helical structures with nonlinearly increasing rotation angles have been studied. The fracture behavior of a 3D-printed helicoidal recursive (HR) composite with nonlinear rotation angle-based layups was investigated by performing quasistatic three-point bending experiments and simulations. First, the crack propagation paths during the loading of the samples were observed, and the critical deformation displacements and fracture toughness were calculated. It was found that the crack path that propagated along the soft phase increased the critical failure displacement and toughness of the samples. Then, the deformation and interlayer stress distribution of the helical structure under static loading were obtained by finite element simulation. The results showed that the variation in the rotation angle between the layers caused different degrees of shear deformation at the interface between adjacent layers, resulting in different shear stress distributions and thus different crack modes of the HR structures. The mixed-mode I + II cracks induced crack deflection, which slowed the eventual failure of the sample and improved the fracture toughness.


Reverse genetics approaches for hepatitis E virus and related viruses.
The hepatitis E virus (HEV) is the causative agent of acute and chronic hepatitis in humans. Related viruses have been found in several animal species. Reverse genetics systems (RGSs), which enable the generation of infectious virus from cloned cDNA by transfection of cultured cells or intrahepatic injection into laboratory animals, have been developed for HEV genotypes 1, 3, 4, 5 and 7 as well as for avian HEV and rat HEV. However, low virus recovery rates and slow replication in cell cultures are observed for most of the HEV types. Nevertheless, the RGSs enabled the site-directed mutagenesis of single nucleotides, deletion of genome fragments, insertion of sequence tags and a marker gene as well as the generation of chimeric viruses.


Okadaic acid inhibits sister chromatid separation in mammalian cells.
Mitotic HeLa cells were treated with different concentrations of okadaic acid inhibiting phosphatase 2A activity alone or in addition to phosphatase 1 activity. Phosphatase 2A inhibition alone had no visible effect on mitosis, but inhibition of both phosphatase 1 and 2A produced mitotic abnormalities, including inhibition of anaphase mimicking the effect of colchicine. Recovery experiments in okadaic acid-free medium showed formation of diplochromosomes, indicating a failure of sister chromatid separation in the treated mitotic cells. The universality of the phosphatase 1 requirement in sister chromatid separation is discussed.


Cardioprotective radiotherapy: the circadian way.
Radiotherapy (RT) has been established to improve both local control as well as overall survival rates in breast cancer. However, RT especially in left-sided breast cancer also irradiates a portion of the heart. Radiation associated toxicity to the heart assumes significance because of improval in survival of breast cancer patients. A circadian pattern has been reported in the myocardial oxygen demand and myocardial ischaemia with the cardiac tissue being more susceptible to injury between 6 am and noon. Radiation damages blood vessels of all sizes causing an increase in capillary wall permeability and dilatation of vessels leading to the characteristic radiation erythema followed by an inflammatory cell infiltrate. Coronary artery spasm may be the reason behind some cases of sudden death occurring in patients after radiation therapy. Endothelial behaviour also has a circadian variation and vasodilation is significantly attenuated in the morning. Critical coronary artery disease occurs 10-15 years after radiotherapy. Radiation in the morning hours may be one of the associated risk factor. The application of chrono-therapeutics with radiation therapy in carcinoma breast and in other chest wall irradiation, could possibly decrease the radiation associated cardiac toxicity.


Distribution of hypertrophied locus coeruleus projection to adult cerebellum after neonatal 6-hydroxydopamine.
Following treatment as neonates with a high subcutaneous dose of 6-hydroxydopamine (6-OHDA), the projections of the locus coeruleus were mapped in the brains of adult rats. This was done using the technique of unilateral lesions in the nucleus followed by simultaneous determinations of norepinephrine (NE) levels, dopamine-beta-hydroxylase (DBH) activity and synaptosomal [3H]NE uptake in various terminal areas. In particular the cerebellum was subdivided into 3 areas in order to assess any changes from normal in the distribution of the hypertrophied noradrenergic projections here. In vehicle-control rats the lesions resulted in an 80--85% loss of NE in the parietal cortex ipsilateral to the lesion and a 15--20% loss contralaterally. In the control cerebellum the locus coeruleus projection, based upon changes in all 3 markers, is distributed 2/3 ipsilaterally and 1/3 contralaterally with the same pattern present in all 3 subregions. The neonatal 6-OHDA treatment resulted in virtually complete loss of noradrenergic terminals in the cerebral cortex. Following neonatal 6-OHDA treatment cerebellar levels of NE, DBH and [3H]NE uptake increased by between 20--60%, with the smallest increases occurring in [3H]NE uptake. In these rats the locus coeruleus accounted for at least 75--80% of the cerebellar noradrenergic parameters. Unlike control rats however, the lesions in these rats produced only ipsilateral decreases in NE and DBH. On the other hand changes in [3H]NE uptake indicated a normal 2/3 ipsilateral, 1/3 contralateral pattern. It is suggested that two separable events occur in the noradrenergic projection to the cerebellum. The first is the regeneration of an increased number of nerve terminals, or sprouting, and the second is a build-up of synaptic vesicles, or collateral accumulation. The sprouting, judging from the [3H]NE uptake data, occurs with apparently normal distribution, but the accumulation of NE and DBH is confined predominantly to the ipsilaterally projecting axon terminals. This may be the consequence of collateral accumulation resulting from the degeneration of the largely ipsilateral coeruleocortical projection.