Protective effects of Hericium erinaceus mycelium and its isolated erinacine A against ischemia-injury-induced neuronal cell death via the inhibition of iNOS/p38 MAPK and nitrotyrosine.
Hericium erinaceus, an edible mushroom, has been demonstrated to potentiate the effects of numerous biological activities. The aim of this study was to investigate whether H. erinaceus mycelium could act as an anti-inflammatory agent to bring about neuroprotection using a model of global ischemic stroke and the mechanisms involved. Rats were treated with H. erinaceus mycelium and its isolated diterpenoid derivative, erinacine A, after ischemia reperfusion brain injuries caused by the occlusion of the two common carotid arteries. The production of inflammatory cytokines in serum and the infracted volume of the brain were measured. The proteins from the stroke animal model (SAM) were evaluated to determine the effect of H. erinaceus mycelium. H. erinaceus mycelium reduced the total infarcted volumes by 22% and 44% at a concentration of 50 and 300 mg/kg, respectively, compared to the SAM group. The levels of acute inflammatory cytokines, including interleukin-1β, interleukin-6 and tumor necrosis factor á, were all reduced by erinacine A. Levels of nitrotyrosine-containing proteins, phosphorylation of p38 MAPK and CCAAT enhancer-binding protein (C/EBP) and homologous protein (CHOP) expression were attenuated by erinacine A. Moreover, the modulation of ischemia injury factors present in the SAM model by erinacine A seemed to result in the suppression of reactive nitrogen species and the downregulation of inducible NO synthase (iNOS), p38 MAPK and CHOP. These findings confirm the nerve-growth properties of Hericium erinaceus mycelium, which include the prevention of ischemic injury to neurons; this protective effect seems to be involved in the in vivo activity of iNOS, p38 MAPK and CHOP.


Mutual A domain interactions in the force sensing protein von Willebrand factor.
The von Willebrand factor (VWF) is a glycoprotein in the blood that plays a central role in hemostasis. Among other functions, VWF is responsible for platelet adhesion at sites of injury via its A1 domain. Its adjacent VWF domain A2 exposes a cleavage site under shear to degrade long VWF fibers in order to prevent thrombosis. Recently, it has been shown that VWF A1/A2 interactions inhibit the binding of platelets to VWF domain A1 in a force-dependent manner prior to A2 cleavage. However, whether and how this interaction also takes place in longer VWF fragments as well as the strength of this interaction in the light of typical elongation forces imposed by the shear flow of blood remained elusive. Here, we addressed these questions by using single molecule force spectroscopy (SMFS), Brownian dynamics (BD), and molecular dynamics (MD) simulations. Our SMFS measurements demonstrate that the A2 domain has the ability to bind not only to single A1 domains but also to VWF A1A2 fragments. SMFS experiments of a mutant [A2] domain, containing a disulfide bond which stabilizes the domain against unfolding, enhanced A1 binding. This observation suggests that the mutant adopts a more stable conformation for binding to A1. We found intermolecular A1/A2 interactions to be preferred over intramolecular A1/A2 interactions. Our data are also consistent with the existence of two cooperatively acting binding sites for A2 in the A1 domain. Our SMFS measurements revealed a slip-bond behavior for the A1/A2 interaction and their lifetimes were estimated for forces acting on VWF multimers at physiological shear rates using BD simulations. Complementary fitting of AFM rupture forces in the MD simulation range adequately reproduced the force response of the A1/A2 complex spanning a wide range of loading rates. In conclusion, we here characterized the auto-inhibitory mechanism of the intramolecular A1/A2 bond as a shear dependent safeguard of VWF, which prevents the interaction of VWF with platelets.


Laser-induced effects in different biological samples.
Experiments were carried out on cancerous HeLa cells and blood serum using a double integrating sphere and a He-Ne laser to investigate the optical properties and cellular effects due to photodynamic therapy (PDT). In the first experiment, HeLa cells were exposed to Photofrin at concentrations of 0, 10, 20, 30, 50 and 112.4 microg/ml at an irradiance of 0.2 W/cm(2) using diode laser light. Using a confocal microscope, cell debris and morphological changes in HeLa cells were recorded at different Photofrin concentrations. The results showed cell debris in HeLa cells at the highest concentration of Photofrin. In a second experiment, photobleaching was observed in HeLa cells in the presence of various concentrations of 5-aminolaevulinic acid ranging from 0-50 microg/ml. There was progressive degradation of the 635 nm peak during continuous laser irradiation at an irradiance of 0.2 W/cm(2). We conclude that cells demonstrating high initial fluorescence undergo bleaching at a faster rate than those with lower fluorescence. Finally in a third experiment, cancerous and noncancerous blood serum was irradiated at an irradiance of 0.1 W/cm(2) using a He-Ne laser in conjunction with a double integrating sphere system. Forward and back scattering of normal and malignant serum showed an exponential decrease in fluorescence amplitude. The results indicate that there is notable amplitude difference between malignant and normal blood serum with malignant blood serum showing decreased scattering. These results have important implications for photodiagnosis and photodynamic therapy.


Comparison of analgesic efficacy and fetal effects between transdermal administration of fentanyl and intramuscular administration of buprenorphine in pregnant sheep.
To compare analgesic efficacy and fetal effects between transdermal administration of fentanyl and IM administration of buprenorphine in pregnant sheep. 12 healthy pregnant ewes. Before study initiation, each ewe was confirmed pregnant with a single fetus between 113 and 117 days of gestation. Ewes were randomly assigned to receive buprenorphine (0.01 mg/kg, IM, q 8 h for 48 hours beginning 1 hour before anesthesia induction; n = 6) or fentanyl (a combination of transdermal fentanyl patches sufficient to deliver a dose of 2 μg of fentanyl/kg/h applied between the dorsal borders of the scapulae 24 hours before anesthesia induction; 6). Ewes were anesthetized and underwent a surgical procedure to instrument the fetus with an arterial catheter and place a catheter in utero for collection of amniotic fluid samples. Physiologic variables and behavioral changes indicative of pain were assessed, and amniotic fluid and blood samples from ewes and fetuses were collected for determination of drug concentrations at predetermined times. Both protocols provided acceptable postoperative analgesia with no adverse effects observed in the ewes or fetuses. Compared with the buprenorphine protocol, the fentanyl protocol induced more profound analgesia, decreased the requirement for isoflurane during surgery, and was associated with a shorter anesthesia recovery time. Fetal indices did not differ significantly between the 2 analgesic protocols. Results indicated that both protocols provided acceptable analgesia. However, the fentanyl protocol was superior in regard to the extent of analgesia induced, inhalant-sparing effects, and anesthesia recovery time.


Structure-function of cytochromes P450 and flavin-containing monooxygenases: implications for drug metabolism.
This article is a report on a symposium held at Experimental Biology '98 in San Francisco, California. Recent developments in site-directed mutagenesis, computer-modeling, and mechanistic analysis of cytochromes P450 and flavin-containing monooxygenases are described. A unifying theme is the elaboration of general approaches for understanding and predicting the function of individual forms of these enzymes. A related goal is the production of soluble forms of mammalian cytochromes P450 for X-ray crystallography.


Network design for telemedicine--e-health using satellite technology.
Over the last decade various international Information and Communications Technology networks have been created for a global access to high-level medical care. OP 2000 has designed and validated the high-end interactive video communication system WinVicos especially for telemedical applications, training of the physician in a distributed environment, teleconsultation and second opinion. WinVicos is operated on a workstation (WoTeSa) using standard hardware components and offers a superior image quality at a moderate transmission bandwidth of up to 2 Mbps. WoTeSa / WinVicos have been applied for IP-based communication in different satellite-based telemedical networks. In the DELTASS-project a disaster scenario was analysed and an appropriate telecommunication system for effective rescue measures for the victims was set up and evaluated. In the MEDASHIP project an integrated system for telemedical services (teleconsultation, teleelectro-cardiography, telesonography) on board of cruise ships and ferries has been set up. EMISPHER offers an equal access for most of the countries of the Euro-Mediterranean area to on-line services for health care in the required quality of service. E-learning applications, real-time telemedicine and shared management of medical assistance have been realized. The innovative developments in ICT with the aim of realizing a ubiquitous access to medical resources for everyone at any time and anywhere (u-Health) bear the risk of creating and amplifying a digital divide in the world. Therefore we have analyzed how the objective needs of the heterogeneous partners can be joined with the result that there is a need for real integration of the various platforms and services. A virtual combination of applications serves as the basic idea for the Virtual Hospital. The development of virtual hospitals and digital medicine helps to bridge the digital divide between different regions of the world and enables equal access to high-level medical care. Pre-operative planning, intra-operative navigation and minimally-invasive surgery require a digital and virtual environment supporting the perception of the physician. As data and computing resources in a virtual hospital are distributed over many sites the concept of the Grid should be integrated with other communication networks and platforms.


Gonadotropin-independent precocious puberty ("testotoxicosis"): influence of maturational status on response to ketoconazole.
Three boys, 5.0-7.4 yr of age, with clinical and biochemical features consistent with familial gonadotropin-independent precocious puberty ("testotoxicosis") were treated with the antifungal drug ketoconazole in a dose of 200 mg every 8 h. Serum testosterone levels fell significantly within 24 h in all three patients, with reciprocal changes in serum 17-hydroxyprogesterone, consistent with inhibition of C17-20 lyase activity. However, after 1-3 months of continuing treatment, an "escape" phenomenon occurred, characterized by progressive increases in serum LH, FSH, and testosterone concentrations. Furthermore, for the first time, a marked pubertal-type LH response was found after GnRH administration in contrast to the absent response consistently found in all patients before ketoconazole therapy. Combination treatment with ketoconazole and the GnRH analog buserelin resulted in restoration of pituitary and gonadal hormone concentrations to the upper range of normal for prepubertal children. We hypothesize that the advanced state of maturation of the boys in this study [mean bone age, 13.2 +/- 0.8 (+/- SE) yr] was the major contributing factor to this escape, with the set-point of the GnRH pulse generator (gonadostat) functioning at the adult level of sensitivity. Such escape may limit the successful use of ketoconazole alone. These data together with evidence of normal testosterone responses to hCG during ketoconazole therapy indicate that in testotoxicosis, pituitary and gonadal receptors are functionally intact, with appropriate negative feedback relationships.


Assessing the magnitude and uncertainties of the burden of selected diseases attributable to extreme heat and extreme precipitation under a climate change scenario in Michigan for the period 2041-2070.
Extreme heat (EH) and extreme precipitation (EP) events are expected to increase with climate change in many parts of the world. Characterizing the potential future morbidity and mortality burden of EH and EP and associated costs, as well as uncertainties in the estimates, can identify areas for public health intervention and inform adaptation strategies. We demonstrate a burden of disease and uncertainty assessment using data from Michigan, USA, and provide approaches for deriving these estimates for locations lacking certain data inputs. Case-crossover analysis adapted from previous Michigan-specific modeling was used to characterize the historical EH-mortality relationship by county poverty rate and age group. Historical EH-associated hospitalization and emergency room visit risks from the literature were adapted to Michigan. In the U.S. Environmental Protection Agency's BenMAP software, we used a novel approach, with multiple spatially-varying exposures, to estimate all non-accidental mortality and morbidity occurring on EH days (EH days; days where maximum temperature 32.2-35 C or > 35 C) and EP days. We did so for two time periods: the "historical" period (1971-2000), and the "projected" period (2041-2070), by county. The rate of all non-accidental mortality associated with EH days increased from 0.46/100,000 persons historically to 2.9/100,000 in the projected period, for 240 EH-attributable deaths annually. EH-associated ED visits increased from 12/100,000 persons to 68/100,000 persons, for 7800 EH-attributable emergency department visits. EP-associated ED visits increased minimally from 1.7 to 1.9/100,000 persons. Mortality and morbidity were highest among those aged 65+ (91% of all deaths). Projected health costs are dominated by EH-associated mortality ($280 million) and EH-associated emergency department visits ($14 million). A variety of sources contribute to a moderate-to-high degree of uncertainty around the point estimates, including uncertainty in the magnitude of climate change, population composition, baseline health rates, and exposure-response estimates. The approach applied here showed that health burden due to climate may significantly rise for all Michigan counties by midcentury. The costs to health care and uncertainties in the estimates, given the potential for substantial attributable burden, provide additional information to guide adaptation measures for EH and EP.


Suppression of carbonic anhydrase III in rat liver by a dioxin-related toxic compound, coplanar polychlorinated biphenyl, 3, 3',4,4',5-pentachlorobiphenyl.
A coplanar polychlorinated biphenyl, 3,3',4,4',5-pentachlorobiphenyl (PenCB), significantly suppresses the expression of rat liver carbonic anhydrase III (CAIII), an enzyme which has recently been suggested to prevent from H(2)O(2)-inducible apoptosis. Marked changes in the CAIII levels of liver cytosol were observed in rats following doses of PenCB ranging from 0.5 to 25 mg/kg body weight and maximum suppression was observed at a dose of 10 mg/kg. Northern analysis revealed that the level of CAIII mRNA in rat liver was dramatically reduced by PenCB treatment while only weak suppression was observed in pair-fed controls. Two AU-rich elements, considered as a destabilizing signal of mRNA, were found in the 3'-untranslated region of CAIII sequenced after reverse transcription-PCR and 3'-rapid amplification of the cDNA end. Dramatic decrease of CAIII in rat liver by PenCB could account for the suppression of the defense system for oxidative stress.


Essential contractual language for medical necessity in children.
The previous policy statement from the American Academy of Pediatrics, "Model Language for Medical Necessity in Children," was published in July 2005. Since that time, there have been new and emerging delivery and payment models. The relationship established between health care providers and health plans should promote arrangements that are beneficial to all who are affected by these contractual arrangements. Pediatricians play an important role in ensuring that the needs of children are addressed in these emerging systems. It is important to recognize that health care plans designed for adults may not meet the needs of children. Language in health care contracts should reflect the health care needs of children and families. Informed pediatricians can make a difference in the care of children and influence the role of primary care physicians in the new paradigms. This policy highlights many of the important elements pediatricians should assess as providers develop a role in emerging care models.


[Features of cholecalciferol hydroxylation in the liver of rats in conditions of D-hypervitaminosis and activity of alpha-tocopherol].
It is shown, that hepatocytes contain two (microsomal and mitochondrial) vitamin D3 25-hydroxylase enzymes, which differ as to their activity and function with maximal activity at different concentrations to substrate, namely at 15 microM and 100 microM of vitamin D3, accordingly. Activity of vitamin D3 25-hydroxylase enzymes of hepatocytes is regulated by cholecalciferol and alpha-tocopherol. The general and microsomal vitamin D3 25-hydroxylase enzymes activity of hepatocytes is lowered, but mitochondrial isoform is increased under D-hypervitaminosis conditions. Vitamin E increases microsomal vitamin D3 25-hydroxylase activity and decreases mitochondrial isoform activity of rats hepatocytes under D-hypervitaminosis conditions. It is established that D-hypervitaminosis is accompanied by expressed hypercalcemia and hyperphosphatemia, by decreased contents of mineral components in the bone tissue and high activity of alkaline phosphatase in the blood serum. The physiological doses of vitamin E under these conditions normalized the mineral metabolism, contents of calcium, phosphates and activity of alkaline phosphatase isoform in the blood serum.


Signs and symptoms of temporomandibular disorders in women and men.
Women are more likely to present temporomandibular disorders (TMD); however, studies comparing genders in Brazilian samples are rare. To analyze the proportion of men and women, as well as the association between gender and age, problem duration, and TMD symptoms in patients admitted to an university clinic for treatment. Interview and assessment data of protocols from 1,000 patients diagnosed with TMD were collected and analyzed and then divided into two groups, male (n = 177) and female (n = 823). The exploratory analysis was based on contingency tables and χ2 test was carried out. Subsequently, the logistic regression model was used and the odds ratios (OR) concerning the evaluated comparisons were calculated. Females were more prevalent in the sample, and mean ages and TMD duration were similar between the groups, with higher occurrence in young adults (19 to 40 years old). The OR values showed an association between the female gender and the signs/symptoms of pain in the temporomandibular joint, pain in the facial muscles, neck and shoulders, headache, fatigue in the muscles of mastication, otologic symptoms, and dysphonia. Women had two times higher chances of presenting these symptoms than men. In the sample of Brazilian patients with TMD, the number of women who presented a higher prevalence of painful symptoms was greater, followed by otologic symptoms and complaints of dysphonia. The prevalence of joint noise was similar in both studied groups.


Effects of Trehalose on Lipid Membranes under Rapid Cooling using All-Atom and Coarse-Grained Molecular Simulations.
We investigate the effect of the cryopreservative α-α-trehalose on a model 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid membrane undergoing cooling from 350 to 250 K using all-atom (AA) and coarse-grained (CG) molecular dynamics simulation. In the AA simulations, we find that the addition of trehalose alters the Lα (liquid crystalline) to Pβ (ripple) phase transition, suppressing the major domain of the Pβ phase and increasing the degree of leaflet interdigitation (the minor domain) which yields a thinner membrane with a higher area per lipid. Calculation of dihedral angle distributions for the lipid tails shows a greater fraction of gauche angles in the Pβ phase as trehalose concentration is increased, indicating that trehalose increases lipid disorder in the membrane. In contrast, the CG simulations transition directly from the Lα to the Lβ (gel) phase upon cooling without exhibiting the Pβ phase (likely due to increased lipid mobility in the CG system). Even so, the CG simulations show that the addition of trehalose clearly suppresses the Lα to Lβ phase transition, demonstrating that trehalose increases lipid disorder at low temperatures for the CG system, similar to the AA. Analysis using a two-state binding model provides net affinity coefficients between trehalose and the membrane as well as trehalose partition coefficients between the membrane interface and the bulk solution for both the AA and CG systems.


Reappraisal of human CYP isoforms involved in imipramine N-demethylation and 2-hydroxylation: a study using microsomes obtained from putative extensive and poor metabolizers of S-mephenytoin and eleven recombinant human CYPs.
Cytochrome P450 (CYP) involved in the two major pathways of imipramine (IMI) was reappraised using human liver microsomes phenotyped for S-mephenytoin 4'-hydroxylation in vitro and 11 recombinant human CYP isoforms. Individual Eadie-Hoffstee plots for IMI N-demethylation and 2-hydroxylation showed a monophasic profile in microsomes obtained from three putative S-mephenytoin poor metabolizer (PM) livers, whereas the plots gave a biphasic relationship (except for one case in 2-hydroxylation) in those from the three extensive metabolizer (EM) livers. Effects of CYP-selective inhibitor/substrate probes on the two metabolic reactions were examined at the two IMI concentrations (2 and 400 microM) with microsomes obtained from the two PM and three EM livers. S-mephenytoin inhibited IMI N-demethylation by 50% at the low concentration in microsomes from the EM livers with no discernible effect on this pathway in those from the PM livers. Furafylline inhibited the N-demethylation by about 60% at the low and high substrate concentrations in microsomes from both the EM and PM livers. Quinidine abolished the 2-hydroxylation at the low and high concentrations in microsomes from both the EM and the PM livers. Among the recombinant human CYPs, CYP2C19, 2C18, 2D6, 1A2, 3A4 and 2B6 in rank order catalyzed the N-demethylation, whereas CYP2D6, 2C19, 1A2, 2C18 and 3A4 catalyzed the 2-hydroxylation. The Km values obtained from recombinant CYP2C19 and 1A2 approximated those of the high- and low-affinity components from human liver microsomes for IMI N-demethylation, respectively. For IMI 2-hydroxylation, the respective Km values obtained from recombinant CYP2D6 and 2C19 were close to those of the high- and low-affinity components from human liver microsomes. Our human liver microsomal study using the near-therapeutic IMI concentration (2 microM) suggests that 1) CYP2C19 and 1A2 are involved in the N-demethylation and the 2-hydroxylation is mediated exclusively by CYP2D6 and partially by CYP2C19 in the EM livers, and 2) CYP1A2 and 2D6 play a major role in IMI N-demethylation and 2-hydroxylation, respectively, in the PM livers. Our recombinant human CYP isoform study, in general, supports this conclusion.


Uroplakin I: a 27-kD protein associated with the asymmetric unit membrane of mammalian urothelium.
The luminal surface of mammalian urothelium is covered with numerous plaques (also known as the asymmetric unit membrane or AUM) composed of semi-crystalline, hexagonal arrays of 12-nm protein particles. Despite the presumed importance of these plaques in stabilizing the urothelial surface during bladder distention, relatively little is known about their protein composition. Using a mouse mAb, AE31, we have identified a 27-kD protein that is urothelium-specific and is differentially expressed in superficial umbrella cells. This protein (pI approximately 5.8) partitions into the detergent phase during Triton X-114 phase separation. Pulse-chase experiments using cultured bovine urothelial cells showed that this protein is synthesized as a 32-kD precursor that is processed through a 30-kD intermediate, to the mature 27-kD form. In cytoplasmic vesicles containing immature AUM, the AE31 epitope is detected in patches on the cytoplasmic side, but in mature, apical AUM it is detected exclusively on the luminal side. This suggests an unusual translocation of the AE31 epitope during AUM maturation; more data are required, however, to substantiate this interpretation. Immunoaffinity purification of the 27-kD protein results in the copurification in approximately molar ratio of a 15-kD protein, as well as a small and variable amount of a 47-kD protein. Immunoblotting data indicate that these three proteins are immunologically distinguishable. This copurified 15-kD protein is relative basic (pI approximately 8.0). Like the 27-kD protein, it is urothelium-specific and is present mainly in the umbrella cells. Together, our data indicate that a 27-kD protein is urothelial plaque-associated (uroplakin I). Based on complex formation data, we provisionally name the 15-kD protein uroplakin II; additional data will be required to determine whether this and the 47-kD protein are integral parts of AUM. The identification of these AUM-associated and -related proteins, plus the availability of a culture system capable of synthesizing and processing some of these molecules, offer new opportunities for studying the detailed structure, assembly, and function of asymmetrical unit membrane.


Selective cytotoxicity of purified homologues of tunicamycin on transformed BALB/3T3 fibroblasts.
The selective cytotoxicity of tunicamycin homologues against SV40-transformed 3T3 cells (SV40-3T3) was examined. Incubation of 3T3 or virally transformed 3T3 cells with four different homologues (A1, A2, B1, and B2 at 0.1 to 0.25 microgram/ml) caused detachment and death of transformed cells after 1 to 3 days, while the nontransformed cells were almost unaffected. Cytotoxicity against nontransformed cells occurred only when higher doses (at least 5-fold) of A2-, B1-, and B2-tunicamycins were used. In contrast, these homologues inhibited proliferation of 3T3 cells, even when doses of 0.5 microgram/ml were used. These cytotoxic effects are dose dependent, and maximal cytotoxicity of each homologue is achieved at a different concentration in each cell type. These results indicate that tunicamycin homologues have selective cytotoxicity against transformed cells. Incorporation of [3H]mannose into acid-precipitable macromolecules synthesized by transformed cells was strongly inhibited (70 to 75%) by A1- and B2-tunicamycins at 0.01 to 0.05 microgram/ml, while incorporation by 3T3 cells was not affected. At higher concentrations of the above tunicamycins (0.5 to 1 microgram/ml), [3H]mannose incorporation by both 3T3 and SV40-3T3 cells was inhibited more than 95%. In contrast, the effect of these tunicamycin homologues on protein synthesis in 3T3 and SV40-3T3 fibroblasts was less pronounced since the incorporation of amino acids was inhibited by approximately 20%. Very little inhibition of amino acid incorporation occurred when 3T3 or SV40-3T3 cells were treated with B2-tunicamycin. However, A1-tunicamycin inhibited [3H]proline incorporation and slightly increased [3H]tyrosine incorporation into cell layers of 3T3 cells. Examination of secreted proteins synthesized by these cells on sodium dodecyl sulfate:polyacrylamide gel electrophoresis revealed that both 3T3 and SV40-3T3 cells treated with homologues produced partially glycosylated macromolecules, such as procollagen and fibronectin, and failed to convert procollagen to collagen. Tunicamycin homologues also inhibited the N-acetylglucosamine-1-phosphate transferase activity found in microsomes prepared from 3T3 and virally transformed 3T3 fibroblasts. The data presented indicate that the cytotoxic activity of purified homologues of tunicamycin against transformed fibroblasts might be due to the selective inhibition of glycosylation and to the differences in the membrane solubilities of the homologues.


White-coat hypertension during coronary computed tomography angiography is associated with higher coronary atherosclerotic burden.
White-coat hypertension (WCH) is a prevalent entity, which has been associated with an increased cardiovascular risk. Assess whether WCH is associated with a higher coronary atherosclerotic burden, evaluated by coronary computed tomography angiography (CCTA) and coronary artery calcium (CAC) scoring. A total of 1362 patients who performed CCTA and simultaneous CAC for the assessment of coronary artery disease (CAD) were prospectively enrolled in a single-center registry and divided into three groups: (A) patients with normal blood pressure (BP) (n=386); (B) patients with WCH (n=174; without a history of hypertension or antihypertensive medication, but with systolic BP ≥140 and/or diastolic BP ≥90 mmHg before examination acquisition); and (C) patients with hypertension (n=802). The following coronary atherosclerotic markers were evaluated: CAC above the 50th percentile (CAC>p50), prevalence of CAD (any plaque), and obstructive CAD (plaque with>50% stenosis). Patients with WCH had a higher coronary atherosclerotic burden compared with patients with normal BP for all markers (30.5 vs. 19.4%, P=0.004 for CAC>p50; 50.6 vs. 36.8%, P=0.002 for CAD, any plaque; and 13.8 vs. 8.3%, P=0.045 for obstructive CAD). On multivariate analysis, WCH was an independent predictor of a CAC>p50 [odds ratio (OR) 1.563, 95% confidence interval 1.018-2.400, P=0.041], but not of the presence of CAD (any plaque) (OR 1.335, P=0.169) or obstructive CAD (OR 1.376, P=0.301). In this registry of patients, WCH was an independent predictor of a CAC above the p50. It was also associated with higher other markers of coronary atherosclerotic burden, such as the presence of CAD on CCTA, compared with patients with normal BP.


Determination of ciprofloxacin, the major metabolite of enrofloxacin, in milk by isopotential fluorimetry.
A new method for the determination of ciprofloxacin, the major metabolite of enrofloxacin, for concentrations between 20 and 200 ng/mL by means of matrix isopotential synchronous fluorescence spectrometry and derivative techniques is proposed. This new method is useful for the determination of compounds in samples with unknown background fluorescence, such as ciprofloxacin in whey, without the need of tedious preseparation. The determination was performed in an ethanol/water medium (20% v/v) at pH 4.8, provided by adding a sodium acetate/acetic acid buffer solution. Since enrofloxacin is widely used as an antibacterial agent in veterinary medicine, the method was successfully applied to the determination of its main metabolite in milk. An exhaustive statistical analysis has been developed to all calibration graphs. This treatment includes robust regression such as least median of squares, which also detects outliers and leverage points. The overall least-squares regression has been applied to find the more exact straight line that fits the experimental data. The error propagation has been considered to calculate the detection limit and the repeatability of the method.


Solvent-annealing-induced nanowetting in templates: towards tailored polymer nanostructures.
We study the solvent-annealing-induced nanowetting in templates using porous anodic aluminum oxide membranes. The morphology of polystyrene and poly(methyl methacrylate) nanostructures can be controlled, depending on whether the swollen polymers are in the partial or complete wetting regimes, which are characterized by the spreading coefficient. When the swollen polymers are in the partial wetting regime, polymers wet the nanopores by capillary action, resulting in the formation of polymer nanorods. When the swollen polymers are in the complete wetting regime, polymers form wetting layers in the nanopores, resulting in the formation of polymer nanotubes. The solubility parameters of polymers and solvents are also used to predict the wetting behavior of swollen polymers in cylindrical geometry.


The A-cluster in subunit beta of the acetyl-CoA decarbonylase/synthase complex from Methanosarcina thermophila: Ni and Fe K-edge XANES and EXAFS analyses.
The acetyl-CoA decarbonylase/synthase (ACDS) complex catalyzes the cleavage of acetyl-CoA in methanogens that metabolize acetate to CO(2) and CH(4), and also carries out acetyl-CoA synthesis during growth on one-carbon substrates. The ACDS complex contains five subunits, among which beta possesses an Ni-Fe-S active-site metal cluster, the A-cluster, at which reaction with acetyl-CoA takes place, generating an acetyl-enzyme species poised for C-C bond cleavage. We have used Ni and Fe K fluorescence XANES and EXAFS analyses to characterize these metals in the ACDS beta subunit, expressed as a C-terminally shortened form. Fe XANES and EXAFS confirmed the presence of an [Fe(4)S(4)] cluster, with typical Fe-S and Fe-Fe distances of 2.3 and 2.7 A respectively. An Fe:Ni ratio of approximately 2:1 was found by Kalphabeta fluorescence analysis, indicating 2 Ni per [Fe(4)S(4)]. Ni XANES simulations were consistent with two distinct Ni sites in cluster A, and the observed spectrum could be modeled as the sum of separate square planar and tetrahedral Ni sites. Treatment of the beta subunit with Ti(3+) citrate resulted in shifts to lower energy, implying significant reduction of the [Fe(4)S(4)] center, along with conversion of a smaller fraction of Ni(II) to Ni(I). Reaction with CO in the presence of Ti(3+) citrate generated a unique Ni XANES spectrum, while effects on the Fe-edge were not very different from the reaction with Ti(3+) alone. Ni EXAFS revealed an average Ni coordination of 2.5 S at 2.19 A and 1.5 N/O at 1.89 A. A distinct feature at approximately 2.95 A most likely results from Ni-Ni interaction. The methanogen beta subunit A-cluster is proposed to consist of an [Fe(4)S(4)] cluster bridged to an Ni-Ni center with one Ni in square planar geometry coordinated by 2 S + 2 N and the other approximately tetrahedral with 3 S + 1 N/O ligands. The electronic consequences of two distinct Ni geometries are discussed.


Paraoxonase (PON1 and PON3) Polymorphisms: Impact on Liver Expression and Atorvastatin-Lactone Hydrolysis.
Atorvastatin δ-lactone, a major, pharmacologically inactive metabolite, has been associated with toxicity. In a previous study we showed that polymorphisms of UGT1A3 influence atorvastatin δ-lactone formation. Here we investigated the reverse reaction, atorvastatin δ-lactone hydrolysis, in a human liver bank. Screening of microarray data revealed paraoxonases PON1 and PON3 among 17 candidate esterases. Microsomal δ-lactone hydrolysis was significantly correlated to PON1 and PON3 protein (r(s) = 0.60; r(s) = 0.62, respectively; P < 0.0001). PON1 and PON3 were strongly correlated to each other (r(s) = 0.60) but PON1 was shown to be more extensively glycosylated than PON3. In addition a novel splice-variant of PON3 was identified. Genotyping of 40 polymorphisms within the PON-locus identified PON1 promoter polymorphisms (-108T > C, -832G > A, -1741G > A) and a tightly linked group of PON3 polymorphisms (-4984A > G, -4105G > A, -1091A > G, -746C > T, and F21F) to be associated with changes in atorvastatin δ-lactone hydrolysis and expression of PON1 but not PON3. However, carriers of the common PON1 polymorphisms L55M or Q192R showed no difference in δ-lactone hydrolysis or PON expression. Haplotype analysis revealed decreased δ-lactone hydrolysis in carriers of the most common haplotype *1 compared to carriers of haplotypes *2, *3, *4, and *7. Analysis of non-genetic factors showed association of hepatocellular and cholangiocellular carcinoma with decreased PON1 and PON3 expression, respectively. Increased C-reactive protein and γ-glutamyl transferase levels were associated with decreased protein expression of both enzymes, and increased bilirubin levels, cholestasis, and presurgical exposure to omeprazole or pantoprazole were related to decreased PON3 protein. In conclusion, PON-locus polymorphisms affect PON1 expression whereas non-genetic factors have an effect on PON1 and PON3 expression. This may influence response to therapy or adverse events in statin treatment.


Screening of Brain Tumors Using Functional Connectivity Patterns of Steady-State Visually Evoked Potentials.
Abstract Objective: The irregular growth and proliferation of cells in the brain and skull is named brain tumor, which is a special serious type of tumor due to its location. Nowadays, brain tumor is diagnosed by using imaging techniques such as computed tomography, positron emission tomography, single photon emission computed tomography, infrared imaging, magnetic resonance imaging (MRI), and functional MRI. However, these methods are not affordable to be used as screening tests. In contrast, electroencephalography has the ability of measuring biological signals, and it is a cost-effective and non-dangerous tool, being feasible to be used for screening purposes. The aim of this research was to evaluate the possibility of brain tumor detection, using functional connectivity features extracted from steady-state visually evoked potentials of eight brain tumor patients and four healthy control participants. Methods: For this purpose, after preprocessing, phase lag index was calculated as a functional connectivity measure. Afterward, four of the channels were chosen as the selected nodes, based on the highest number of strong connectivity (top 5%) as well as the most significant ones. The selected nodes were O1, O2, P3, and P4. As a final step, node strength was calculated for these selected nodes, which was used to classify the samples in the two groups, using Naïve Bayes, discriminant analysis (DA), k-nearest neighbors, support vector machines, and logistic regression methods. Results: The highest accuracy of 89.6% was obtained by using a DA classifier. This result shows that, in fact, brain tumor has the ability of changing brain functional connectivity. Conclusion: As the physiological alterations may occur sooner than the anatomical ones in tumor onset, detection of these alterations may be a useful measure for early diagnosis of this disease. This is still a primary study, but with the possibility of leading to further research, which can lead to the development of a method for the screening and early detection of brain tumor. Impact statement Imaging modalities are often used to diagnose brain tumors. However, these approaches are cost prohibitive for screening tests. Electroencephalography, a method that measures biological signals from the brain, on the other hand, is cost-effective, non-dangerous, and non-invasive, making it an ideal screening tool. The goal of this study is to analyze whether functional connectivity patterns taken from Steady-State Visually Evoked Potentials may be used to detect brain tumors at early stages. Since physiological changes occur before anatomical ones in tumor development, monitoring these alterations could be a useful tool to screen high-risk individuals, and also to an early diagnosis of the disease.


G3139 (oblimersen) may inhibit prostate cancer cell growth in a partially bis-CpG-dependent non-antisense manner.
G3139 is an 18-mer phosphorothioate oligodeoxyribonucleotide, which is targeted to the initiation codon region of the bcl-2mRNA. Although treatment of PC3 prostate cancer cells with G3139, which contains two CpG motifs, causes a dramatic decrease in bcl-2 protein expression after 3 days, it did not result in significant cellular apoptosis, as it does in many other cell lines. The absence of apoptosis was demonstrated by the absence of pro-caspase 3 cleavage products and of Annexin V cell surface expression. In addition, ATP production and the mitochondrial membrane potential DeltaPsim were preserved. Despite this, G3139 significantly inhibited the rate of cellular proliferation in complete media and blocked cloning in soft agar. G4232, a variant of G3139 that down-regulates bcl-2 expression to the same extent but has both CpG cytidines C5 methylated, was only minimally antiproliferative. A series of mismatched G3139-related oligomers were synthesized that could also substantially down-regulate bcl-2 protein expression, but only if the CpG motifs were preserved, demonstrating the presence of additional non-antisense mechanisms. G3139 caused production of reactive oxygen species in growth-arrested cells and oxidation of nuclear guanosine to 8-hydroxy-2'-deoxyguanosine, as determined by 1F7 monoclonal antibody staining. Bromodeoxyuridine incorporation studies demonstrated that G3139 induced a G1-S entry block and an intra-S-phase block in PC3 cells that persisted as long as 3 days. This finding coincides with the observation that expression of several proteins encoded by S-phase genes, including c-myb and poly(ADP-ribose) polymerase, were significantly reduced. These results illustrate the complexity of the mechanism of action of G3139 in PC3 cells.


[Biology of papillomavirus infections. I. General characteristics].
Papillomaviruses are pathogens which induce cutaneous and mucosal lesions in man and in many animal species. The characterization of these viruses was rather low, because viral infection cannot be fully reproduced in cell culture. The development of molecular biology techniques in the 1970s permitted to establish the remarkable plurality of the viruses, the tissue specificity and pecular pathogenicity linked to the type. Studies of the genome organization, the gene expression regulation and the protein characterization gave many informations leading to understand the mechanisms of viral-related carcinogenesis, especially the role of HPV16, the major risk factor for the development of squamous cervical carcinoma.


[Calcitonin--contra-insulin hormone].
The review contains literature data about calcitonin's participation in the glucose metabolism. The analyses of retrospective and current sources of information about contra-insulin effect of calcitonin on the glucose metabolism has been revealed. Some arguments of antagonistic calcitonin effect to insulin under the pre-receptor, cell level and liver were done. The impairment of glucose metabolism under calcitonin is established--hyperglycemia, insulin resistance and glucose intolerance. A calcitonin diabetogenic effect and its role as a "risk factor" in the development of metabolic syndrome and diabetic mellitus are discussed.


Overlap of familial Mediterranean fever and hyper-IgD syndrome in an Arabic kindred.
Hyperimmunoglobulinemia D Syndrome (HIDS) has rarely been reported in Arabs. Moreover, the simultaneous presence of mutations in MEFV and MVK segregating in the same family is exceptional. We report an Arabic girl presenting since the age of 8-years with two patterns of recurrent episodes of fever, and associated with a spectrum of clinical features suggestive of overlap between familial Mediterranean fever (FMF) and HIDS. Her 19-year old brother presented since the age of 1 year with prolonged episodes of fever and was diagnosed with HIDS at the age of 7 years based on clinical features and homozygosity for p.V377I mutation in MVK. Shorter episodes of fever and abdominal pain more consistent with FMF ensued since the age of 17 years. Genetic testing done for both patients and all other family members revealed simultaneous presence of mutations in MEFV and MVK but with a variable clinical spectrum ranging from asymptomatic to severe manifestations. Both of our patients are homozygous for p.V377I MVK mutation; the girl is a compound heterozygote for p.E148Q/p.P369S/p.R408G and p.E167D/p.F479L MEFV mutations whereas the brother is a compound heterozygote for p.E148Q/p.P369S/p.R408G and p.M680I MEFV mutations. The clinical implications of having more than one mutation in different genes of monogenic autoinflammatory diseases in the same individual are not clear but may explain atypical clinical manifestations such as the overlap features of both FMF and HIDS in this family.


Binding of the recombinant proteinase inhibitor eglin c, of the soybean Bowman-Birk proteinase inhibitor and of its chymotrypsin and trypsin inhibiting fragments to Leu-proteinase, the leucine specific serine proteinase from spinach (Spinacia oleracea L.) leaves: thermodynamic study.
The effect of pH and temperature on the apparent association equilibrium constant (Ka) for the binding of the recombinant proteinase inhibitor eglin c (eglin c), of the soybean Bowman-Birk proteinase inhibitor (BBI) and of its chymotrypsin and trypsin inhibiting fragments (F-C and F-T, respectively) to Leu-proteinase, the leucine specific serine proteinase from spinach (Spinacia oleracea L.) leaves, has been investigated. On lowering the pH from 9.5 to 4.5, values of Ka (at 21 degrees C) for complex formation decrease thus reflecting the acidic pK-shift of the hystidyl catalytic residue from approximately 6.9, in the free Leu-proteinase, to approximately 5.1, in the enzyme: inhibitor adducts. At pH 8.0, values of the apparent thermodynamic parameters for the proteinase:inhibitor complex formation are: Leu-proteinase:eglin c-Ka = 2.2 x 10(11) M-1, delta G degree = -64 kJ/mol, delta H degree = +5.9 kJ/mol, and delta S degree = +240 kJ/molK; Leu-proteinase:BBI-Ka = 3.2 x 10(10) M-1, delta G degree = -59 kJ/mol, delta H degree = +8.8 kJ/mol, and delta S degree = +230 J/molK; and Leu-proteinase:F-C-Ka = 1.1 x 10(6) M-1, delta G degree = -34 kJ/mol, delta H degree = +18 J/mol, and delta S degree = +180 J/molK (values of Ka, delta G degree and delta S degree were obtained at 21.0 degrees C; values of delta H degree were temperature-independent over the range explored, i.e. between 10.0 degrees C and 40.0 degrees C).(ABSTRACT TRUNCATED AT 250 WORDS)


Investigation of factors associated with treatment-seeking in Southern Chinese with orofacial pain.
To investigate key factors associated with treatment-seeking for orofacial pain symptoms in community-dwelling adult Chinese people in Hong Kong. A cross-sectional study was conducted using a Hong Kong Hospital Authority Family Medicine Clinic as the sampling frame. People aged 35 to 70 years with recent orofacial pain symptoms participated. Standard questions were asked about orofacial pain symptoms and characteristics in the previous month, Oral Health Impact Profile (OHIP-14), General Health Questionnaire (GHQ-12), Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) depression/nonspecific physical symptoms (NPS) scales, sleep and illness behavior measures, and questions on pain disability, professional treatment-seeking, pain medication usage, and dental attendance were administered prior to a standard clinical assessment. Multiple logistic regression with a forward stepwise selection method was used for data analysis. Two hundred people with orofacial pain symptoms participated in the definitive study. Twenty-seven percent had sought professional advice. The majority had consulted a medical practitioner (66.7%) or dentist (40.7%), 16.7% had consulted a traditional Chinese medicine (TCM) practitioner, and the majority (60%) had taken pain medication. There was no significant association between different orofacial pain diagnoses and professional treatment-seeking (P = .602). Four independent factors were significantly related to an increased likelihood of treatment-seeking (P < .05): more frequent dental attendance for check-up (odds ratio [OR] > 3), time when pain was experienced during the past month (days) (OR > 5), multiple pain symptoms (OR = 4.99), and use of TCM when ill (OR = 3.31). Professional treatment-seeking for orofacial pain was low in this ethnic group. The strongest predictor of treatment-seeking for orofacial pain was an increase in the number of days when pain was experienced during the past month. Key factors associated with treatment-seeking should be considered when formulating community health outreach programs. A further validation study with a larger sample size is recommended to confirm the present findings.


Obesity, Galectin-3, and Incident Heart Failure: The ARIC Study.
Background Laboratory data suggest obesity is linked to myocardial inflammation and fibrosis, but clinical data are limited. We aimed to examine the association of obesity with galectin-3, a biomarker of cardiac inflammation and fibrosis, and the related implications for heart failure (HF) risk. Methods and Results We evaluated 8687 participants (mean age 63 years; 21% Black) at ARIC (Atherosclerosis Risk in Communities) Visit 4 (1996-1998) who were free of heart disease. We used adjusted logistic regression to estimate the association of body mass index (BMI) categories with elevated galectin-3 (≥75th sex-specific percentile) overall and across demographic subgroups, with tests for interaction. We used Cox proportional hazards models to assess the combined associations of galectin-3 and BMI with incident HF (through December 31, 2019). Higher BMI was associated with higher odds of elevated galectin-3 (odds ratio [OR], 2.32; 95% CI, 1.88-2.86) for severe obesity ([BMI ≥35 kg/m2] versus normal weight [BMI 18.5-<25 kg/m2]). There were stronger associations of BMI with elevated galectin-3 among women versus men and White versus Black participants (both P-for-interaction <0.05). Elevated galectin-3 was similarly associated with incident HF among people with and without obesity (HR, 1.49; 95% CI, 1.18-1.88; and HR, 1.71; 95% CI, 1.38-2.11, respectively). People with severe obesity and elevated galectin-3 had >4-fold higher risk of HF (HR, 4.19; 95% CI, 2.98-5.88) than those with normal weight and galectin-3 <25th percentile. Conclusions Obesity is strongly associated with elevated galectin-3. Additionally, the combination of obesity and elevated galectin-3 is associated with marked HF risk, underscoring the importance of elucidating pathways linking obesity with cardiac inflammation and fibrosis.


Surgical treatment of coledochal cyst associated with an aberrant posterior hepatic duct: report of a case and brief literature review.
Choledochal cysts (CCs) are rare congenital cystic or fusiform dilatations of the biliary tree that can involve the extrahepatic and/or intrahepatic biliary tree. We report a case of huge type I CC associated with an aberrant posterior hepatic duct. A 52-year-old man presented with a 3-week history of upper right abdominal pain and jaundice and serologic sign of obstructive jaundice. Ultrasonography (US), magnetic resonance cholangiopancreatography and endoscopic retrograde cholangiopancreatography were performed with the diagnosis of CC type I according to the classification of Alonso-Lej and Todani-Watanabe. The indication for surgical resection was posed. The cyst was completely resected and the biliary tract was reconstructed with a double hepatico-jejunostomy using the same Roux limb, since during the surgical dissection a before unrecognized anatomical variation of the right biliary tree (aberrant posterior hepatic duct at VI-VII segment) was identified. The diagnosis of CC is often difficult and US and magnetic resonance cholangiopancreatography are necessary to definite biliary dilatation. Endoscopic retrograde cholangiopancreatography should be the most definitive and reliable procedure for the diagnosis and treatment of bilio-pancreatic disorders. Gold standard treatment is surgery (bilio-jejunostomy) and frozen-section histology should be performed to rule out the presence of cancer. In conclusion, surgery is the gold standard for the treatment of CC type I and does not depend on the age of patients, based on a substantial lifetime risk of developing cholangiocarcinoma. Preoperative study is mandatory to assess the biliary tree morphology and to research any anatomical variation.


Interactive molecular dynamics in virtual reality for accurate flexible protein-ligand docking.
Simulating drug binding and unbinding is a challenge, as the rugged energy landscapes that separate bound and unbound states require extensive sampling that consumes significant computational resources. Here, we describe the use of interactive molecular dynamics in virtual reality (iMD-VR) as an accurate low-cost strategy for flexible protein-ligand docking. We outline an experimental protocol which enables expert iMD-VR users to guide ligands into and out of the binding pockets of trypsin, neuraminidase, and HIV-1 protease, and recreate their respective crystallographic protein-ligand binding poses within 5-10 minutes. Following a brief training phase, our studies shown that iMD-VR novices were able to generate unbinding and rebinding pathways on similar timescales as iMD-VR experts, with the majority able to recover binding poses within 2.15 Å RMSD of the crystallographic binding pose. These results indicate that iMD-VR affords sufficient control for users to carry out the detailed atomic manipulations required to dock flexible ligands into dynamic enzyme active sites and recover crystallographic poses, offering an interesting new approach for simulating drug docking and generating binding hypotheses.


Petit mal status.
Petit mal status is a heterogeneous clinical syndrome of nonconvulsive status epilepticus. The EEG accompaniment is likewise heterogeneous. Petit mal status occurs at all ages. The characteristics of this syndrome are quite nonspecific and consist of (a) behavioral changes, usually associated with lethargy, slowness, and decreased mental function, (b) abnormal generalized continuous or nearly continuous epileptiform EEG activity, and (c) absence of gross tonic-clonic activity or highly lateralized clonic activity. These criteria do not distinguish petit mal status from complex partial status. Clinical evidence for abruptness of recovery and EEG evidence for localization are required for this distinction, an important factor in therapeutic decisions.


Inflammation-Induced Protein Unfolding in Airway Smooth Muscle Triggers a Homeostatic Response in Mitochondria.
The effects of airway inflammation on airway smooth muscle (ASM) are mediated by pro-inflammatory cytokines such as tumor necrosis factor alpha (TNFα). In this review article, we will provide a unifying hypothesis for a homeostatic response to airway inflammation that mitigates oxidative stress and thereby provides resilience to ASM. Previous studies have shown that acute exposure to TNFα increases ASM force generation in response to muscarinic stimulation (hyper-reactivity) resulting in increased ATP consumption and increased tension cost. To meet this increased energetic demand, mitochondrial O2 consumption and oxidative phosphorylation increases but at the cost of increased reactive oxygen species (ROS) production (oxidative stress). TNFα-induced oxidative stress results in the accumulation of unfolded proteins in the endoplasmic reticulum (ER) and mitochondria of ASM. In the ER, TNFα selectively phosphorylates inositol-requiring enzyme 1 alpha (pIRE1α) triggering downstream splicing of the transcription factor X-box binding protein 1 (XBP1s); thus, activating the pIRE1α/XBP1s ER stress pathway. Protein unfolding in mitochondria also triggers an unfolded protein response (mtUPR). In our conceptual framework, we hypothesize that activation of these pathways is homeostatically directed towards mitochondrial remodeling via an increase in peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC1α) expression, which in turn triggers: (1) mitochondrial fragmentation (increased dynamin-related protein-1 (Drp1) and reduced mitofusin-2 (Mfn2) expression) and mitophagy (activation of the Phosphatase and tensin homolog (PTEN)-induced putative kinase 1 (PINK1)/Parkin mitophagy pathway) to improve mitochondrial quality; (2) reduced Mfn2 also results in a disruption of mitochondrial tethering to the ER and reduced mitochondrial Ca2+ influx; and (3) mitochondrial biogenesis and increased mitochondrial volume density. The homeostatic remodeling of mitochondria results in more efficient O2 consumption and oxidative phosphorylation and reduced ROS formation by individual mitochondrion, while still meeting the increased ATP demand. Thus, the energetic load of hyper-reactivity is shared across the mitochondrial pool within ASM cells.


Computerized image-searching method for finding correct patients for misfiled chest radiographs in a PACS server by use of biological fingerprints.
We have developed an automated image-searching method based on biological fingerprints for identifying correct patients in misfiled chest radiographs in a picture archiving and communication system (PACS) server. We used five biological fingerprints including distinctive anatomic structures in a misfiled chest radiograph of an unknown patient to find another image of the same patient stored with correct patient information in a PACS server. The correlation values were determined for the corresponding biological fingerprints in all images in the image server. The correlation indices as a measure of the overall similarity of the two images were determined from the summation of five correlation values and the combination of correlation values with the weighting factors. Finally, the correct patient was identified automatically by the image with the highest correlation index. By use of the summation of five correlation values as the correlation index, 78.0% (156/200) of the 200 patients for misfiled images were correctly identified in the database. When we applied the weighting factors for each biological fingerprint to determine the correlation index, the performance in identifying the correct patient was improved to 87.5% (175/200). An additional 5.0% (10/200) of images were included in the Top 10 ranking of the correlation index in the database. These cases could be identified manually by radiology personnel. We conclude that the automated image-searching method based on biological fingerprints with weighting factors would be useful for identification of the correct patient in the case of misfiled chest radiographs in a PACS server.


DNA-RNA chimeric hammerhead ribozyme to transforming growth factor-beta(1) mRNA inhibits the exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats.
The purpose of this study was to develop DNA-RNA chimeric hammerhead ribozyme against transforming growth factor-beta(1) (TGF-beta(1)) mRNA as a gene therapy agent for arterial proliferative diseases. A 38-base hammerhead ribozyme against rat TGF-beta(1) mRNA, to produce cleavage at the GUC sequence at nucleotide 825 according to the secondary structure of rat TGF-beta(1) mRNA was designed. To enhance its stability, we synthesized a DNA-RNA chimeric ribozyme with two phosphorothioate linkages at the 3'-terminal. We also synthesized a mismatch ribozyme with single base change in the catalytic loop region as a control. These ribozymes were delivered into rat vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats by lipofectin-mediated transfection, and their biological effects were investigated. According to in vitro cleavage studies, the synthetic ribozyme can cleave the synthetic substrate RNA into two RNA fragments. Chimeric ribozyme significantly inhibited DNA synthesis in VSMC from SHR but not in cells from WKY rats. Mismatch ribozyme showed only a little effect on growth of VSMC from SHR. Chimeric ribozyme significantly inhibited proliferation of VSMC from SHR; in contrast, the proliferation of VSMC from WKY rats was significantly increased by this chimeric ribozyme. Mismatch ribozyme did not affect proliferation of VSMC from either rat strain. Chimeric hammerhead ribozyme to rat TGF-beta(1) dose-dependently inhibited TGF-beta(1) mRNA expression detected by reverse transcription and polymerase chain reaction analysis in VSMC from both rat strains. Chimeric hammerhead ribozyme to rat TGF-beta(1) also dose-dependently inhibited TGF-beta(1) protein production detected by Western blot analysis. The present results demonstrated that our designed DNA-RNA chimeric hammerhead ribozyme to TGF-beta(1) mRNA might be a useful gene therapy agent for hypertensive vascular diseases.


A unique variant of a homeobox gene related to Drosophila cut is expressed in mouse testis.
Mammalian homologues of the Drosophila cut homeobox gene encode transcriptional repressors that are involved in tissue-specific and developmental gene regulation. We examined the expression of a murine cut homologue (Cux-1) in the adult mouse. In many somatic tissues, Cux-1 was expressed as a 13-kb transcript. However, the highest expression of Cux-1 was in the testis, where a unique 2.4-kb splice variant was identified. Less abundant transcripts of 5 kb, 6.5 kb, and 8.5 kb were also detected only in the testis. The nucleotide sequence of the 2.4-kb Cux-1 transcript was identical to the 13-kb transcript in the region of overlap, but the testis-specific transcript encoded a truncated protein that contained only one Cut repeat in addition to the Cut-related homeodomain. Studies of mice homozygous for the atrichosis (at/at) mutation suggested that the 2.4-kb transcript was expressed in germ cells in the testis. In situ hybridization verified that Cux-1 was transiently expressed in post-meiotic germ cells at the round spermatid stage. Immunoblot analysis of nuclear extracts showed that the testis-specific Cux-1 transcripts encoded a 55-kDa protein. These results demonstrate that multiple products of a cut-related homeobox gene are expressed in the testis. The highly restricted pattern of expression of Cux-1 in the testis suggests that it may be involved in regulation of postmeiotic gene expression.


Motor Subtype as a Predictor of Future Working Memory Performance in Idiopathic Parkinson's Disease.
Parkinson's disease is a progressive neurodegenerative disorder associated with reduced spatial and verbal working memory ability. There are two established motor subtypes of PD, tremor dominant (TD) and postural instability and gait difficulty (PIGD). This study used structural equation modelling to explore the longitudinal relationship between the two subtypes and working memory assessed at a 2-year follow-up. The study comprised 84 males and 30 females (N = 114), aged between 39 and 85 (M = 64.82, SD = 9.23) with confirmed PD. There was no significant relationship between motor subtype at Time 1 and working memory at Time 2. Postural symptom severity at Time 1 predicted Time 2 spatial working memory for the PIGD subtype (p = .011) but not the TD subtype. Tremor symptoms were not associated with Time 2 working memory in either subtype. Predictive significance of Time 1 postural symptoms only in the PIGD subtype suggests an interaction between symptom dominance (subtype) and symptom severity that future subtyping should consider. This study demonstrates a predictive relationship between postural difficulties and working memory performance assessed at a 2-year follow-up. Establishing physical symptoms as predictors of cognitive change could have significant clinical importance.


Plant and animal PR1 family members inhibit programmed cell death and suppress bacterial pathogens in plant tissues.
A role for programmed cell death (PCD) has been established as the basis for plant-microbe interactions. A functional plant-based cDNA library screen identified possible anti-PCD genes, including one member of the PR1 family, designated P14a, from tomato. Members of the PR1 family have been subject to extensive research in view of their possible role in resistance against pathogens. The PR1 family is represented in every plant species studied to date and homologues have been found in animals, fungi and insects. However, the biological function of the PR1 protein from plants has remained elusive in spite of extensive research regarding a role in the response of plants to disease. Constitutive expression of P14a in transgenic tomato roots protected the roots against PCD triggered by Fumonisin B1, as did the human orthologue GLIPR1, indicating a kingdom crossing function for PR1. Tobacco plants transformed with a P14a-GFP fusion construct and inoculated with Pseudomonas syringae pv. tabaci revealed that the mRNA was abundant throughout the leaves, but the fusion protein was restricted to the lesion margins, where cell death and bacterial spread were arrested. Vitus vinifera grapes expressing the PR1 homologue P14a as a transgene were protected against the cell death symptoms of Pierce's disease. A pull-down assay identified putative PR1-interacting proteins, including members of the Rac1 immune complex, known to function in innate immunity in rice and animal systems. The findings herein are consistent with a role of PR1 in the suppression of cell death-dependent disease symptoms and a possible mode of action.


Constitutive Models for the Tensile Behaviour of TRM Materials: Literature Review and Experimental Verification.
In recent years, the scientific community has focused its interest on innovative inorganic matrix composite materials, namely TRM (Textile Reinforced Mortar). This class of materials satisfies the need of retrofitting existing masonry buildings, by keeping the compatibility with the substrate. Different recent studies were addressed to improve the knowledge on their mechanical behaviour and some theoretical models were proposed for predicting the tensile response of TRM strips. However, this task is complex due to the heterogeneity of the constituent materials and the stress transfer mechanism developed between matrix and fabric through the interface in the cracked stage. This paper presents a state-of-the-art review on the existing constitutive models for the tensile behavior of TRM composites. Literature experimental results of tensile tests on TRM coupons are presented and compared with the most relevant analytical models proposed until now. Finally, a new experimental study is presented and its results are used to further verify the reliability of the literature expressions.


Epidemiological risk factors and phylogenetic affinities of Sarcocystis infecting village chickens and pigs in Peninsular Malaysia.
The intake of food and water containing the Sarcocystis parasite has been linked to a number of outbreaks worldwide, including Malaysia. Nevertheless, the lack of surveys and epidemiological data on Sarcocystis infections in Malaysia makes it difficult to estimate its occurrence in humans and animals. A cross-sectional study was conducted to determine the prevalence of Sarcocystis and the risk factors associated with infection among village chickens and pigs reared under different farm managements in Peninsular Malaysia. Phylogenetic trees were constructed using partial fragments of the 18S rRNA gene and ITS1 sequences. In the present study, 680 sera samples were collected from village chickens (n=250) and commercial pigs (n=433) and anti-Sarcocystis antibodies were screened using the enzymelinked immunosorbent assays (ELISA) kit. At the animal level, the prevalence of Sarcocystis was 9.2% (95% CI: 5.92-13.48) and at the farm level, it was 64.0% (95% CI: 42.52-82.03) in village chickens. The animal-level seroprevalence of Sarcocystis for pigs was 3.7% (95% CI: 2.13-5.93) and 36.8% (95% CI: 16.29-61.64) at the farm-level. Polymerase Chain Reaction (PCR) was conducted on meat samples from various parts of village chickens (n=250) consisting of brain, heart, lung, and pectoralis muscle tissues, and pork (n=121) consisting of intercostal muscle, diaphragm, and tongue. Sarcocystis DNA was detected in 6.4% (95% CI: 4.60-11.60) of village chicken samples but zero in pork samples. A total of 11 unique Sarcocystis haplotypes were isolated from these tissue samples. Multivariable logistic regression analysis of the putative risk factors showed a statistically significant association between Sarcocystis infection in pigs and uncovered storage of feed. Although no zoonotic Sarcocystis was isolated in this study, we reported the first discovery of S. wenzeli in Malaysia.


Copeptin levels in patients with coronary artery ectasia.
The etiology and pathophysiology of coronary artery ectasia (CAE) has not been fully elucidated. A rapid rise in plasma copeptin has been observed in cardiovascular diseases, stroke, sepsis, and shock. This increase has diagnostic and prognostic value. The aim of this study was to investigate whether copeptin has a relationship with CAE. This observational prospective study was carried out between October 2012 and March 2013 in the cardiology catheter laboratory with the inclusion of 44 subjects with a diagnosis of CAE and 44 age- and gender-matched individuals with normal coronary arteries. Blood samples obtained from the patients were stored at -70 °C until analysis and copeptin levels in sera were measured by ELISA. This study comprised 88 study participants, among whom 44 were patients meeting ectasia criteria [mean age: 58.0 ± 11.5 years; 59% (n = 26) male], and 44 were control subjects with angiographically normal coronary anatomy [mean age: 49.2 ± 10.1 years; 30% (n = 13) male]. Copeptin levels were similar between the groups (p > 0.05). In addition, there was no correlation between copeptin levels and CAE. In this study, it is examined that copeptin levels were not elevated in CAE patients.


An efficient sequential approach to tracking multiple objects through crowds for real-time intelligent CCTV systems.
Efficiency and robustness are the two most important issues for multiobject tracking algorithms in real-time intelligent video surveillance systems. We propose a novel 2.5-D approach to real-time multiobject tracking in crowds, which is formulated as a maximum a posteriori estimation problem and is approximated through an assignment step and a location step. Observing that the occluding object is usually less affected by the occluded objects, sequential solutions for the assignment and the location are derived. A novel dominant color histogram (DCH) is proposed as an efficient object model. The DCH can be regarded as a generalized color histogram, where dominant colors are selected based on a given distance measure. Comparing with conventional color histograms, the DCH only requires a few color components (31 on average). Furthermore, our theoretical analysis and evaluation on real data have shown that DCHs are robust to illumination changes. Using the DCH, efficient implementations of sequential solutions for the assignment and location steps are proposed. The assignment step includes the estimation of the depth order for the objects in a dispersing group, one-by-one assignment, and feature exclusion from the group representation. The location step includes the depth-order estimation for the objects in a new group, the two-phase mean-shift location, and the exclusion of tracked objects from the new position in the group. Multiobject tracking results and evaluation from public data sets are presented. Experiments on image sequences captured from crowded public environments have shown good tracking results, where about 90% of the objects have been successfully tracked with the correct identification numbers by the proposed method. Our results and evaluation have indicated that the method is efficient and robust for tracking multiple objects (>or= 3) in complex occlusion for real-world surveillance scenarios.


Degloving combined with structural trauma at the digital level: functional coverage with fascial free flaps.
Major trauma to the digit that includes skin degloving and tendon loss, as well as fractures, nerve segmental loss, and devascularization can render that digit chronically painful, stiff, insensate, and a detriment to the overall function of the hand to the point that such digits may be candidates for ray resection. Large wounds may exceed the ability to achieve coverage by regional pedicle flaps. When the wound is restricted to the level of the digit, the narrow cylindrical shape and the ability to flex and fold make an absolute requirement for a very thin, as well as supple, flap a requirement that cutaneous free flaps cannot meet. This series reports on 12 male patients with major degloving and structural trauma at the digit level in which bone, tendon, and nerve reconstructions were covered with free fascial flaps and immediate split-thickness skin grafts, followed by early rehabilitation. All patients achieved sufficient functional recovery so that subsequent surgeries were not indicated.


Fruit position within the canopy affects kernel lipid composition of hazelnuts.
The aim of this research was to study the variability in kernel composition within the canopy of hazelnut trees. Kernel fresh and dry weight increased linearly with fruit height above the ground. Fat content decreased, while protein and ash content increased, from the bottom to the top layers of the canopy. The level of unsaturation of fatty acids decreased from the bottom to the top of the canopy. Thus, the kernels located in the bottom layers of the canopy appear to be more interesting from a nutritional point of view, but their lipids may be more exposed to oxidation. The content of different phytosterols increased progressively from bottom to top canopy layers. Most of these effects correlated with the pattern in light distribution inside the canopy. The results of this study indicate that fruit position within the canopy is an important factor in determining hazelnut kernel growth and composition. © 2017 Society of Chemical Industry.


Glycation and glycoxidation of low-density lipoproteins by glucose and low-molecular mass aldehydes. Formation of modified and oxidized particles.
Patients with diabetes mellitus suffer from an increased incidence of complications including cardiovascular disease and cataracts; the mechanisms responsible for this are not fully understood. One characteristic of such complications is an accumulation of advanced glycation end-products formed by the adduction of glucose or species derived from glucose, such as low-molecular mass aldehydes, to proteins. These reactions can be nonoxidative (glycation) or oxidative (glycoxidation) and result in the conversion of low-density lipoproteins (LDL) to a form that is recognized by the scavenger receptors of macrophages. This results in the accumulation of cholesterol and cholesteryl esters within macrophages and the formation of foam cells, a hallmark of atherosclerosis. The nature of the LDL modifications required for cellular recognition and unregulated uptake are poorly understood. We have therefore examined the nature, time course, and extent of LDL modifications induced by glucose and two aldehydes, methylglyoxal and glycolaldehyde. It has been shown that these agents modify Arg, Lys and Trp residues of the apoB protein of LDL, with the extent of modification induced by the two aldehydes being more rapid than with glucose. These processes are rapid and unaffected by low concentrations of copper ions. In contrast, lipid and protein oxidation are slow processes and occur to a limited extent in the absence of added copper ions. No evidence was obtained for the stimulation of lipid or protein oxidation by glucose or methylglyoxal in the presence of copper ions, whereas glycolaldehyde stimulated such reactions to a modest extent. These results suggest that the earliest significant events in this system are metal ion-independent glycation (modification) of the protein component of LDL, whilst oxidative events (glycoxidation or direct oxidation of lipid or proteins) only occur to any significant extent at later time points. This 'carbonyl-stress' may facilitate the formation of foam cells and the vascular complications of diabetes.


Chemiluminescence of lipid vesicles supplemented with cytochrome c and hydroperoxide.
The increase in light emission of hydroperoxide-supplemented cytochrome c observed on addition of lipid vesicles was related to the degree of unsaturation of the fatty acids of the phospholipids: dipalmitoyl phosphatidylcholine was without effect, whereas dioleoyl phosphatidylcholine and soya-bean phosphatidylcholine enhanced chemiluminescence 2- and 3-fold respectively. Effects on light-emission were similar to those on O2 uptake. The chemiluminescence of the present system was sensitive to cyanide and to the radical trap 2,5-di-t-butylquinol, indicating a catlytic activity of cytochrome c and the presence of free-radical species respectively. Lipid-vesicle enhanced chemiluminescence showed different kinetic behaviours, apparently depending on unsaturation: three phases are described for soya-bean phosphatidylcholine, whereas only one phase was present in mixtures containing dipalmitoyl and dioleoyl phospholipids. Chemiluminescence of lipid vesicles supplemented with cytochrome c and hydroperoxide showed similar kinetic patterns with H2O2 and primary (ethyl) and tertiary (t-butyl and cumene) hydroperoxides. Participation of singlet molecular oxygen, mainly on the phase III of chemiluminescence, is suggested by the increase of light-emission by 1,4-diazabicyclo[2.2.2]-octane as well as by data from spectral analysis.


Characteristics of Ba2+-stimulated insulin release with special reference to pancreatic beta-cells sensitized by cyclic AMP.
Ca2+-dependent processes are activated by Ba2+ in a variety of biological systems. When Ca2+ was replaced by equimolar amounts of Ba2+ there was a marked increase in insulin secretion from beta-cell-rich pancreatic islets microdissected from ob/ob-mice. At both 3 and 20 mM glucose Ba2+ stimulated insulin release in a concentration-dependent manner, being less stimulatory at high concentrations. The stimulatory effect of Ba2+ on insulin release is similar to that of Ca2+ in being more pronounced and reached at lower concentrations when the beta-cells were sensitized by cyclic AMP. However, both glucose oxidation and utilization were suppressed when Ca2+ was replaced by equimolar amounts of Ba2+. Ba2+-stimulated insulin release resembled physiological secretion initiated by Ca2+ in being inhibited by L-epinephrine, pentobarbital and a low oxygen tension.


Natural HLA class I polymorphism controls the pathway of antigen presentation and susceptibility to viral evasion.
HLA class I polymorphism creates diversity in epitope specificity and T cell repertoire. We show that HLA polymorphism also controls the choice of Ag presentation pathway. A single amino acid polymorphism that distinguishes HLA-B*4402 (Asp116) from B*4405 (Tyr116) permits B*4405 to constitutively acquire peptides without any detectable incorporation into the transporter associated with Ag presentation (TAP)-associated peptide loading complex even under conditions of extreme peptide starvation. This mode of peptide capture is less susceptible to viral interference than the conventional loading pathway used by HLA-B*4402 that involves assembly of class I molecules within the peptide loading complex. Thus, B*4402 and B*4405 are at opposite extremes of a natural spectrum in HLA class I dependence on the PLC for Ag presentation. These findings unveil a new layer of MHC polymorphism that affects the generic pathway of Ag loading, revealing an unsuspected evolutionary trade-off in selection for optimal HLA class I loading versus effective pathogen evasion.


Treatment and outcome of chronic tenosynovitis in three horses.
Three horses were presented for treatment of chronic infections of the digital flexor tendon sheath. Clinical signs included severe lameness, and heat, pain and swelling of the digital flexor tendon sheath. The horses were treated with surgical lavage of the tendon sheath, systemic and local antibiotics, and analgesics. In each case, resolution of the lameness occurred over weeks to months. Only one horse returned to athletic activity, while the other two became comfortable at pasture. Response to treatment in cases of chronic tenosynovitis may not be as rapid or complete as that reported for acute tendon sheath infections.


Persistence of Bacteriophage Phi 6 on Porous and Nonporous Surfaces and the Potential for Its Use as an Ebola Virus or Coronavirus Surrogate.
The infection of health care workers during the 2013 to 2016 Ebola outbreak raised concerns about fomite transmission. In the wake of the coronavirus disease 2019 (COVID-19) pandemic, investigations are ongoing to determine the role of fomites in coronavirus transmission as well. The bacteriophage phi 6 has a phospholipid envelope and is commonly used in environmental studies as a surrogate for human enveloped viruses. The persistence of phi 6 was evaluated as a surrogate for Ebola virus (EBOV) and coronaviruses on porous and nonporous hospital surfaces. Phi 6 was suspended in a body fluid simulant and inoculated onto 1-cm2 coupons of steel, plastic, and two fabric curtain types. The coupons were placed at two controlled absolute humidity (AH) levels: a low AH of 3.0 g/m3 and a high AH of 14.4 g/m3 Phi 6 declined at a lower rate on all materials under low-AH conditions, with a decay rate of 0.06-log10 PFU/day to 0.11-log10 PFU/day, than under the higher AH conditions, with a decay rate of 0.65-log10 PFU/h to 1.42-log10 PFU/day. There was a significant difference in decay rates between porous and nonporous surfaces at both low AH (P < 0.0001) and high AH (P < 0.0001). Under these laboratory-simulated conditions, phi 6 was found to be a conservative surrogate for EBOV under low-AH conditions in that it persisted longer than Ebola virus in similar AH conditions. Additionally, some coronaviruses persist longer than phi 6 under similar conditions; therefore, phi 6 may not be a suitable surrogate for coronaviruses.IMPORTANCE Understanding the persistence of enveloped viruses helps inform infection control practices and procedures in health care facilities and community settings. These data convey to public health investigators that enveloped viruses can persist and remain infective on surfaces, thus demonstrating a potential risk for transmission. Under these laboratory-simulated Western indoor hospital conditions, we assessed the suitability of phi 6 as a surrogate for environmental persistence research related to enveloped viruses, including EBOV and coronaviruses.


Endogenous sex hormones in men aged 40-80 years.
To determine the relationship between aging, life-style factors and health-related factors and endogenous sex hormone levels. Cross-sectional study of 400 independently living men between 40 and 80 Years of age. After exclusion of subjects who were not physically or mentally able to visit the study center, 400 men were randomly selected from a population-based sample. Total testosterone (TT), bioavailable testosterone (BT) (i.e. not bound to sex hormone-binding globulin (SHBG)), SHBG, estradiol (E(2)) and dehydroepiandrosterone-sulfate (DHEA-S) were investigated for their relationship with age, body mass index (BMI), waist circumference, smoking, physical activity and general health status. Multivariate models using ANCOVA analyses were used to examine the contribution of life-style factors to sex hormone variability. TT, BT and DHEA-S decreased with age; 0.2, 0.7 and 1.2%/Year respectively. SHBG showed an increase with age of 1.1%/Year. No changes with age were found for E(2). General health status modified the association of TT and SHBG with age (P interaction 0.10 and 0.002 respectively). Increased BMI and waist circumference were associated with decreased TT, BT, SHBG and DHEA-S and increased E(2) (all P<0.01). Current smoking, lower alcohol intake and a higher physical activity score were associated with higher TT and SHBG levels. This study showed the important determinants of sex hormones were age, BMI, waist circumference, smoking, general health status and physical activity. Furthermore, it can be concluded that general health status modified the effect between sex hormones and age. For future observational studies it should be taken into account that the above-mentioned determinants may alter the association between sex hormones and diseases and related conditions.


The impact of climate change and glacier mass loss on the hydrology in the Mont-Blanc massif.
The Mont-Blanc massif, being iconic with its large glaciers and peaks of over 4,000 m, will experience a sharp increase in summer temperatures during the twenty-first century. By 2100, the impact of climate change on the cryosphere and hydrosphere in the Alps is expected to lead to a decrease in annual river discharge. In this work, we modelled the twenty-first century evolution of runoff in the Arve river, downstream of Mont-Blanc's French side. For the first time for this region, we have forced a hydrological model with output from an ice-dynamical glacier model and 16 downscaled climate projections, under RCP4.5 and RCP8.5 scenarios. By 2100, under RCP8.5 (high-emission scenario), the winter discharge of the Arve river remains low but is expected to increase by 80% when compared to the beginning of the century. By contrast, the summer season, currently the most important discharge period, will be marked by a runoff decrease of approximately 40%. These changes are almost similar according to a scenario with a lower warming (RCP4.5) and are mostly driven by glacier retreat. These shifts will have significant downstream impacts on water quantity and quality, affecting hydroelectric generation, agriculture, forestry, tourism and aquatic ecosystems.


Topography and Expansion Patterns at the Biofilm-Agar Interface in Bacillus subtilis Biofilms.
Bacterial biofilms are complex microbial communities that are formed on various natural and synthetic surfaces. In contrast to bacteria in their planktonic form, biofilms are characterized by their relatively low susceptibility to anti-microbial treatments, in part due to limited diffusion throughout the biofilm and the complex distribution of bacterial cells within. The virulence of biofilms is therefore a combination of the structural properties and patterns of adhesion that anchor them to their host surface. In this paper, we analyze the topographical properties of Bacillus subtilis' biofilm-agar interface across different growth conditions. B. subtilis colonies were grown to maturity on biofilm-promoting agar-based media (LBGM), under standard and stress-inducing growth conditions. The biofilm-agar interface of the colony-type biofilms was modeled using confocal microscopy and computational analysis. Profilometry data were obtained from the macrocolonies and used for the analysis of the surface topography as it relates to the adhesion modes present at the biofilm-agar interface. Fluorescent microspheres were utilized to monitor the expansion patterns present at the interface between the macrocolonies and the solid growth medium. Contact surface analysis revealed topographical changes that could have a direct effect on the adhesion strength of the biofilm to its host surface, thus affecting its potential susceptibility to anti-microbial agents. The topographical characteristics of the biofilm-agar interface partially define the macrocolony structure and may have significant effects on bacterial survival and virulence.


Distinguishing primary and secondary disorders in studies of alcohol dependence and depression.
This paper is concerned with the relationship between alcohol dependence and depression, and more specifically with the issue of which criteria should be used in distinguishing primary and secondary disorders. The data presented indicate that borderline or subclinical depression frequently precedes the onset of alcohol dependence. This finding is discussed in terms of previous studies, which show that depression is generally secondary to alcohol dependence. Such studies have focused almost exclusively on severe forms of depressive disorder, and, it is argued, used inappropriate criteria to designate the onset of alcohol dependence (e.g. withdrawal phenomena and alcohol-related problems). In contrast to previous studies, the present paper indicates that subclinical episodes of depression should be assessed by clinicians as possible predisposing factors in the development of alcohol dependence. In addition, it is argued that research designed to establish whether alcohol dependence is a primary or secondary disorder when it occurs in the presence of an accompanying psychiatric disorder should measure onset in terms of changes in behaviour such as giving-up existing interests in order to spend time drinking and consuming more alcohol than former companions.


Rare variant association testing in the non-coding genome.
The development of next-generation sequencing technologies has opened-up some new possibilities to explore the contribution of genetic variants to human diseases and in particular that of rare variants. Statistical methods have been developed to test for association with rare variants that require the definition of testing units and, in these testing units, the selection of qualifying variants to include in the test. In the coding regions of the genome, testing units are usually the different genes and qualifying variants are selected based on their functional effects on the encoded proteins. Extending these tests to the non-coding regions of the genome is challenging. Testing units are difficult to define as the non-coding genome organisation is still rather unknown. Qualifying variants are difficult to select as the functional impact of non-coding variants on gene expression is hard to predict. These difficulties could explain why very few investigators so far have analysed the non-coding parts of their whole genome sequencing data. These non-coding parts yet represent the vast majority of the genome and some studies suggest that they could play a major role in disease susceptibility. In this review, we discuss recent experimental and statistical developments to gain knowledge on the non-coding genome and how this knowledge could be used to include rare non-coding variants in association tests. We describe the few studies that have considered variants from the non-coding genome in association tests and how they managed to define testing units and select qualifying variants.


The control of ribonucleic acid synthesis in bacteria. Steady-state content of messenger ribonucleic acid in Escherichia coli M.R.E. 600.
1. The technique of DNA-RNA hybridization was used to follow changes in the amount and average lifetime of unstable messenger RNA in Escherichia coli M.R.E. 600 over a wide range of different growth conditions. The method of analysis was based on the kinetics of incorporation of exogenous labelled nucleic acid bases into the RNA of steadily growing cultures, as described by Bolton & McCarthy (1962). 2. The ratio of the average lifetime of messenger RNA to the mean generation time of E. coli cultures was constant over the temperature range 25-45 degrees C in a given medium, but the constant varied with the nature of the growth medium. For cultures growing in sodium lactate-salts or glucose-salts media the ratio was 0.046+/-0.005 and in enriched broth it was 0.087+/-0.009. Measurements of the amounts of transfer RNA, ribosomal RNA and messenger RNA were also made. The results confirmed earlier reports that the ratio of the amount of messenger RNA to the amount of ribosomes in the cells is virtually constant. On the other hand, the ratio of the amount of transfer RNA to the amount of ribosomal RNA decreased with increasing growth rate at a given temperature. 3. In cultures at temperatures higher than necessary for optimum rates of growth the average lifetime of messenger RNA lengthened in harmony with the increased time required for cell division. It seems that suboptimum growth rates at higher temperatures cannot be explained simply as a combination of increased rates of synthesis and breakdown of messenger RNA with a grossly decreased efficiency of translation. The absolute rate of messenger RNA synthesis was lowered, and its amount in the cells was typical of all other cultures grown at lower temperatures in the same medium. 4. The rate of entry of exogenous labelled uracil into unstable messenger RNA and stable ribosomal RNA was constant in all media at all temperatures in the approximate ratio 1:2. In media supporting a lower rate of growth, e.g. lactate-salts or glucose-salts media, the messenger RNA fraction constituted 2.2+/-0.3% of the total cellular RNA. In enriched broth 3.6+/-0.3% of the total RNA was messenger.


Effect of alendronate on bone mineral density and bone turnover in Thai postmenopausal osteoporosis.
Alendronate has recently been approved for the prevention and treatment of postmenopausal osteoporosis, and its efficacy has been demonstrated in many Western countries. Our present study was performed to evaluate the effect of alendronate on bone mineral density (BMD) and its tolerability in Thais. Eighty postmenopausal women with osteoporosis participated in this study. After giving informed consent, the subjects were randomly allocated either 10 mg alendronate or placebo in a double-blind fashion. All patients received a supplement of 500 mg elemental calcium daily. BMD at the lumbar spine, femoral neck, and distal forearm was measured at baseline and 6 and 12 months after treatment. Biochemical markers of bone resorption were determined at baseline and 6 months after treatment. Baseline characteristics were similar in both alendronate- and placebo-treated groups. Ten subjects discontinued the study. Of 70 subjects, 32 received 10 mg alendronate daily and the remaining subjects received placebo. At 1 year, BMD in the alendronate-treated group had increased from baseline by 9.2%, 4.6%, and 3.1% at lumbar spine, femoral neck, and distal forearm, respectively. These percentages were greater than those in controls (4.1%, 0.6%, and 1.0%, respectively). Urinary N-terminal telopeptide (NTx)-I and serum C-terminal telopeptide (CTx)-I levels decreased in both groups after 6 months of treatment. However, more reduction was demonstrated in the alendronate-treated group (71.9% vs. 28.4%, P < 0.01, and 84.7% vs. 33.1%, P < 0.01, respectively). Compliance with treatment and drug tolerability were good in both alendronate and placebo groups. We concluded that treatment with alendronate 10 mg daily for Thai postmenopausal women with osteoporosis significantly increased BMD at all skeletal sites and reduced biochemical markers of bone resorption. It was well tolerated without any serious side effects.


Antiobesity Effect of Prebiotic Polyphenol-Rich Grape Seed Flour Supplemented with Probiotic Kefir-Derived Lactic Acid Bacteria.
The interaction between prebiotics and probiotics may exert synergistic health benefits. This study investigated the combined effects of polyphenol-rich wine grape seed flour (GSF), a prebiotic, and lactic acid bacteria (LAB) derived from kefir, a probiotic, on obesity-related metabolic disease in high-fat diet (HFD) induced obese (DIO) mice. DIO mice were fed with HFD with 6% microcrystalline cellulose (CON) or HFD supplemented with GSF (5% or 10% GSF), HFD with LAB orally administrated (LAB), or HFD with a combination of GSF and LAB orally administrated (GSF+LAB) for 9 weeks. The vehicle, saline, was also orally administered to the CON and GSF groups. In comparison to CON, all GSF and LAB groups showed a reduction ( P < 0.05) in HF-induced weight gain, liver and adipose tissue weights, plasma lipid concentrations, insulin resistance, and glucose intolerance. The combination of 10% GSF and LAB showed synergistic effects ( P < 0.05) on body weight gain, plasma insulin and total cholesterol concentrations, and cecum propionate contents. Plasma zonulin and cecum propionate concentrations and intestinal FXR gene expression were ( P < 0.05) correlated with body weight gain. A pathway analysis of microarray data of adipose tissue showed that the combination of GSF and LAB affected genes involved in metabolic and immunological diseases, including inflammasome complex assembly ( P < 0.05). In conclusion, a combination of GSF and LAB inhibited HF-induced obesity and inflammation via alterations in intestinal permeability and adipocyte gene expression.


Natural orifice transluminal endoscopic surgery (NOTES): current experience and urologic applications.
Natural orifice transluminal endoscopic surgery (NOTES) has attracted considerable recent attention for its potential to allow traditional abdominal procedures to be performed without a transabdominal incision. With considerable experience in the development and application of minimally invasive techniques, urologists have played a significant role in early experimental NOTES efforts and have contributed to early investigations in human subjects accordingly. However, adoption of these techniques has been limited due to cumbersome endoscopic equipment and concerns regarding peritonitis from failed viscerotomy closure. Experience with use of NOTES in human subjects is limited, and studies comparing NOTES to conventional minimally invasive techniques are lacking. Until adequate endoscopes are developed to facilitate a pure NOTES approach, multiple portals of entry will be necessary to facilitate both urologic and non-urologic reconstructive and extirpative procedures. Our aim is to evaluate NOTES techniques, portals of entry, early clinical experiences, and the application of NOTES to urologic surgery.


Synthesis and 18F-radiolabeling of thymidine AMBF3 conjugates.
In pursuit of 18F-labeled nucleosides for positron emission tomography (PET) imaging, we report on the chemical and radiochemical synthesis of two thymidine (dT) analogs, dT-C5-AMBF3 and dT-N3-AMBF3, that are radiofluorinated by isotope exchange (IEX) and studied as PET imaging agents in mice with tumor xenografts. dT-C5-AMBF3 shows preferential, and tumor-specific, uptake over dT-N3-AMBF3. This work provides a new synthetic method in order to access new nucleoside tracers for PET imaging.