Dynamics of aquaporin-2 serine-261 phosphorylation in response to short-term vasopressin treatment in collecting duct.
We recently identified a novel phosphorylation site, serine-261 (pS261), in the COOH-terminus of the vasopressin-regulated water channel, aquaporin-2 (AQP2). To address whether phosphorylation at this site is regulated by vasopressin, a rabbit polyclonal phospho-specific antibody was generated. Dot blot and immunoblot analysis demonstrated that this antibody specifically recognizes AQP2 phosphorylated at pS261, and that phosphorylation of S256 (pS256), a site already known to be regulated by vasopressin, does not interfere with antibody recognition. Immunohistochemical analysis revealed intense pS261 labeling of inner medullary collecting duct (IMCD) from wild-type mice, while sections from AQP2 knockout animals showed a general absence of labeling. AQP2 pS261 was present in principal cells of all mouse and rat distal tubule segments from the connecting tubule to the terminal IMCD. Co-immunolabeling of collecting duct with phospho-specific and total AQP2 antibodies revealed that pS261 and pS256 have distinct subcellular distributions. Levels of pS256 increased, while the amount of pS261 significantly decreased in freshly isolated rat IMCD samples incubated with 1 nM [deamino-Cys(1),D-Arg(8)]vasopressin for 30 min. Similarly, based on immunohistochemical labeling, the amount of pS261 was reduced in all collecting duct segments of Brattleboro rats treated with [deamino-Cys(1),D-Arg(8)]vasopressin for 2 h. This study reveals a reciprocal change in S256 and S261 phosphorylation in response to short-term vasopressin exposure, suggesting that these residues may serve distinct roles in regulation of AQP2 subcellular distribution and collecting duct water permeability.


Ligand-Directed Reticular Synthesis of Catalytically Active Missing Zirconium-Based Metal-Organic Frameworks.
Zirconium-based metal-organic frameworks (Zr-MOFs) based on edge-transitive nets such as fcu, spn, she, csq, and ftw with diverse potential applications have been widely reported. Zr-MOFs based on the highly connected 6,12-connected alb net, however, remain absent on account of synthetic challenges. Herein we report the ligand-directed reticular syntheses and isoreticular expansion of a series of Zr-MOFs with the edge-transitive alb net from 12-connected hexagonal-prismatic Zr6 nodes and 6-connected trigonal-prismatic linkers, i.e., microporous NU-1600, mesoporous NU-1601, and mesoporous NU-1602. These Zr-MOFs exhibit remarkable activities toward the destruction of a nerve agent (soman) and a nerve agent simulant (DMNP).


Grip strength and finger dexterity across five styles of commercial wrist orthoses.
Five styles of commercial static wrist extensor orthoses were compared to determine whether any style, or styles, afforded better power grip strength or finger dexterity. Because wrist extensor orthoses are intended for use during functional tasks, their influence on hand function is of great importance. Twenty-three right-hand-dominant women without upper extremity dysfunction participated in this crossover study. Dominant-hand finger dexterity and power grip strength were evaluated while wearing each of five commercial orthoses-Kendall-Futuro #33 (Futuro), AliMed Freedom Long (AliMed Long), AliMed Freedom Short (AliMed Short), Smith & Nephew Rolyan D-Ring (Rolyan), and LMB Wrist Rest (LMB)--and while using the dominant hand without an orthosis (free hand). Finger dexterity was assessed with the unimanual subtest of the Purdue Pegboard. Grip strength was assessed with a Jamar hydraulic dynamometer. Four of the study orthoses (Futuro, AliMed Short, Rolyan, and LMB) afforded finger dexterity that did not differ significantly from that of the free hand. The AliMed Long orthosis slowed finger speed when compared with the speeds afforded by both the LMB orthosis and the free hand. The Rolyan orthosis permitted a power grip strength that was not significantly different from the free hand. The other four commercial orthoses reduced grip strength when compared with the strength observed when wearing a Rolyan orthosis and when gripping with a free hand. The five styles of commercial orthoses affect power grip and finger dexterity differently. When power grip or finger dexterity are priorities, differences among the orthoses furnish grounds for initial suggestions, although medical needs and patient preference should be the overriding factors in the final selection of an orthosis.


MR imaging of focal medullary sponge kidney: case report.
We present a case of focal medullary sponge kidney (MSK) that mimicked a renal tumor. Evaluation of a patient with history of macrohematuria revealed a left renal mass of 3-cm diameter. T(1)-weighted magnetic resonance (MR) images revealed a mass of mixed intensity protruding toward the renal sinus. On fat-saturated T(2)-weighted MR images, the lesion's remarkable hyperintensity suggested the presence of an aggregation of tiny cysts. On diffusion-weighted MR images, the mass also demonstrated high intensity, and its apparent diffusion coefficient was partly decreased (1.12 × 10(-3) mm(2)/s). On computed tomography, precontrast images revealed no calcification in the mass. Although slight enhancement was seen in the corticomedullary phase, thick and dense streaks of contrast radiating peripherally were identified in the mass in the excretory phase. Focal MSK was diagnosed. We discuss the potential of MR imaging for diagnosing focal MSK.


The pathology of alcohol use and abuse.
Alcohol is the most widely abused substance in the United States and its pathology is responsible for more pathological conditions than all other forms of drug use combined. Alcohol dependence is associated with a number of adverse individual and societal consequences and high rates of morbidity and mortality. Alcohol use and abuse have a significant pathological effect on the brain, fetus, liver, heart, pancreas, and immune system. Cancer risks have also been attributed to alcohol use and abuse. Assessing acute and chronic alcohol consumption is critical to effective treatment but unfortunately currently available clinical laboratory testing procedures lack the ability to inform alcohol treatment providers about use and abuse.


CO2 laser increases the regenerative capacity of human adipose-derived stem cells by a mechanism involving the redox state and enhanced secretion of pro-angiogenic molecules.
CO2 laser has a beneficial effect on stem cells by mechanisms that are not clearly elucidated. We hypothesize that the effect of fractional CO2 laser on human adipose-derived stem cells (ADSC) could be due to changes in redox homeostasis and secretion of factors contributing to cellular proliferation and angiogenic potential. ADSC incubated in medium containing 0.5 or 10 % FBS were exposed to a single irradiation of a 10,600-nm fractional CO2 laser; non-irradiated ADSC were used as control. Viability/proliferation of ADSC was assessed by MTT assay; the intracellular reactive oxygen species (ROS) levels and the mitochondrial membrane potential (∆Ψm) were determined with DCFH-DA and JC-1 fluorescent probes, respectively. Molecules secreted by ADSC in the medium were determined by ELISA assay, and their capacity to support endothelial tube-like formation by the Matrigel assay. The results showed that compared to controls, ADSC kept in low FBS medium and irradiated with CO2 laser at 9 W exhibited: (a) increased proliferation (∼20 %), (b) transient increase of mitochondrial ROS and the capacity to restore Δψm after rotenone induced depolarization, and (c) augmented secretion in the conditioned medium of MMP-2 (twofold), MMP-9 (eightfold), VEGF (twofold), and adiponectin (∼50 %) that have the capacity to support angiogenesis of endothelial progenitor cells. In conclusion, the mechanisms underlying the benefic effect of CO2 laser on ADSC are the activation of the redox pathways which increases cell proliferation and enhances secretion of angiogenic molecules. These results explain, in part, the mechanisms involved in the increased regenerative potential of CO2 laser-exposed ADSC that could be exploited for clinical applications.


Pharmacokinetics of leucovorin (D,L-5-formyltetrahydrofolate) after intravenous injection and constant intravenous infusion.
The pharmacokinetics of the active and inactive diastereoisomers of leucovorin and its active metabolite, 5-methyltetrahydrofolate (5-CH3-THF) were studied after iv injection of leucovorin in normal human subjects at a dose of 28 mg/m2, and in patients given 500 mg/m2 daily by constant iv infusion for a 5.5 day period. In both studies the plasma half-life (t1/2) of the active isomer, L-formyltetrahydrofolate (CHO-THF), was only 32 to 35 minutes, whereas the inactive isomer, D-CHO-THF had a plasma t 1/2 of 352 to 485 minutes. During constant infusion, the plasma levels reached plateaus of 2.33 and 37.5 microM for L-CHO-THF and D-CHO-THF, respectively. The inactive isomer was cleared from plasma only by urinary excretion of the unchanged drug. The active isomer was also excreted unchanged in the urine but in addition was extensively metabolized to the active metabolite L-5-CH3-THF. The active metabolite achieved a plasma level of 4.85 microM during constant infusion and appeared to have a longer t 1/2 after constant infusion than was observed after iv injection. Furthermore a larger apparent volume of distribution (Vd) of 5-CH3-THF was obtained in the constant infusion study. These findings suggest that constant iv infusion of large doses of leucovorin can considerably expand the intracellular pools of active folate. The consequence of the extensive accumulation of the inactive isomer, D-CHO-THF, is not known. However, the small Vd of D-CHO-THF suggests that it does not extensively accumulate in tissues.


Pleistocene refugia and polytopic replacement of diploids by tetraploids in the Patagonian and Subantarctic plant Hypochaeris incana (Asteraceae, Cichorieae).
We report the phylogeographic pattern of the Patagonian and Subantarctic plant Hypochaeris incana endemic to southeastern South America. We applied amplified fragment length polymorphism (AFLP) and chloroplast DNA (cpDNA) analysis to 28 and 32 populations, respectively, throughout its distributional range and assessed ploidy levels using flow cytometry. While cpDNA data suggest repeated or simultaneous parallel colonization of Patagonia and Tierra del Fuego by several haplotypes and/or hybridization, AFLPs reveal three clusters corresponding to geographic regions. The central and northern Patagonian clusters (approximately 38-51 degrees S), which are closer to the outgroup, contain mainly tetraploid, isolated and highly differentiated populations with low genetic diversity. To the contrary, the southern Patagonian and Fuegian cluster (approximately 51-55 degrees S) contains mainly diploid populations with high genetic diversity and connected by high levels of gene flow. The data suggest that H. incana originated at the diploid level in central or northern Patagonia, from where it migrated south. All three areas, northern, central and southern, have similar levels of rare and private AFLP bands, suggesting that all three served as refugia for H. incana during glacial times. In southern Patagonia and Tierra del Fuego, the species seems to have expanded its populational system in postglacial times, when the climate became warmer and more humid. In central and northern Patagonia, the populations seem to have become restricted to favourable sites with increasing temperature and decreasing moisture and there was a parallel replacement of diploids by tetraploids in local populations.


The limits of lifestyle: re-assessing 'fatalism' in the popular culture of illness prevention.
This paper is concerned with the development of preventive medicine in the field of Coronary Heart Disease. It is based on an in-depth, ethnographic investigation into the popular culture of prophylactic behaviour carried out in South Wales (U.K.) during 1988 and 1989. The focus of the data and analysis presented here is the operation of cultural norms and practices related to the understanding and explanation of the cause and distribution of illness and death from heart ailments. The paper illustrates how the everyday cultural practice of 'lay epidemiology' is involved in accounting for illness misfortune and in assessing the potential benefits of prophylactic behaviour change. A central issue dealt with here is the relationship of lifestyle to environment in the popular understanding of chronic disease. Lay notions of luck, fate, destiny, randomness and chaos in the distribution of heart disease are explored. In conclusion, some implications for health education in this field are put forward.


EEG spectral activity during paradoxical sleep: further evidence for cognitive processing.
Paradoxical sleep (PS), in which periods with (phasic) and without (tonic) rapid eye movements are intermingled, is hypothesized to be related to cognitive processing and dreaming. Based on polysomnographic data from 12 healthy subjects, this study focuses on the spectral differentiation between phasic and tonic periods. Phasic PS periods exhibited decreased theta and alpha power in the posterior brain areas suggesting the interference of visual processing related to dream imagery. Phasic PS periods were also characterized by a shift from beta to gamma activity in frontal, central and occipital areas reflecting specific phasic related activation. Together, these findings bring new evidence for the existence of visual imagery and cognitive processing during phasic PS.


High-speed screening and quantitative SAR analysis of human ABC transporter ABCG2 for molecular modeling of anticancer drugs to circumvent multidrug resistance.
The transport mechanism-based molecular design strategy would provide an effective tool for rationalized chemotherapy against tumors. To develop a platform for molecular modeling to circumvent multidrug resistance, we established new methods of high-speed screening for human ABCG2-drug interactions, quantitative structure-activity relationship (QSAR) analysis, and quantum chemical calculation for lead optimization.


Clinical usefulness of bedside intracranial morphological monitoring: mobile computerized tomography in the neurosurgery intensive care unit. Report of three cases.
The practice of modern neurointensive care is based on the use of multimodality monitoring to respond rapidly to physiological, biochemical, or morphological changes and avoid secondary brain injury. Until recently, one important monitoring method, computerized tomography (CT), has not been available for bedside use. The authors have over 3 years of experience with the routine use of bedside CT scanning and have developed their own method of scanning the patients in their beds. In this report, they describe three illustrative cases in which the mobile CT scanner was of great value in the management of difficult neurosurgical intensive care problems. It is concluded that the availability of bedside morphological monitoring in the neurosurgery intensive care unit is of great help in management and clinical decision making.


The subacromial impingement syndrome. A study of results of treatment with special emphasis on predictive factors and pain-generating mechanisms.
Forty-two patients with subacromial impingement syndrome entered a randomized prospective study comparing open anterior acromioplasty with a physiotherapy regime. The criterion for a successful outcome of treatment was a reduction of the initial pain score of more than 50% using the visual analogue scale (VAS) technique. The evaluation was performed by an independent observer. At the 6-month follow-up, treatment in the surgical group had been successful in 12/21 (57%) patients versus 6/18 (33%) in the physiotherapy group. A one-year evaluation revealed 16/21 (76%) "successes" in the surgical group. A direct and unbiased comparison with the physiotherapy group was not possible at one year, since 13 patients chose surgery after initial physiotherapy. However, in "an intention to treat" analysis outcome at one year was significantly better in those randomized to surgery. We found two standardized, composite, active movements--the "Pour out of a Pot" manoeuvre requiring the emptying of a pot of water, and the "Hand in Neck" manoeuvre--to be of predictive value for the outcome of surgery. When combining three pain-related variables into a criterion for prediction of success, a sensitivity of 78% (p < 0.02) and a specificity of 90% (p < 0.03) were attained. We advance the hypothesis that pain in the impingement syndrome is mainly elicited by comparison of the subacromial bursa. In some patients a traction-responsive pain generator in the supraspinatus tendon may be present as well.


Determination of four carboxylic acid metabolites of felodipine in plasma by high-performance liquid chromatography.
A reversed-phase high-performance liquid chromatography method with ultraviolet detection at 220 nm was developed to determine four carboxylic acid metabolites in plasma following therapeutic doses of the calcium antagonist felodipine. After the addition of an internal standard the analytes were isolated by liquid-liquid and solid-phase extraction. The metabolites were applied to a C2 cartridge in their free acid form, but they were transformed and retained as ion pairs with tetrabutylammonium during a wash with phosphate buffer (pH 7), prior to automated elution and injection by the Varian AASP system onto the analytical C18 column. Using a sample volume of 1 ml of plasma, the lower limit of determination for the metabolites was about 20 nmol/l. The influence of the pH of the mobile phase on the retention time of the metabolites and the structural requirements for the internal standard were studied. The method was applied to plasma samples from four dogs collected after an oral dose of felodipine. The plasma concentration-time profiles of the metabolites gave useful information about the mechanisms by which they were formed and eliminated.


Alive and Well after 25 Years: A Review of Groupthink Research.
This article provides a summary of empirical research on groupthink theory. Groupthink research, including analyses of historical cases of poor group decision making and laboratory tests of groupthink, is reviewed. Results from these two research areas are briefly compared. Theoretical and methodological issues for future groupthink research are identified and discussed. I conclude that groupthink research has had and continues to have considerable heuristic value. A small, but growing, body of empirical literature has been generated. In addition, groupthink research has stimulated a number of theoretical ideas, most of which have yet to be tested. Copyright 1998 Academic Press.


Changes in incidence and indications of tonsillectomy and adenotonsillectomy, 1970-2005.
To investigate 35-year epidemiological trends in tonsillectomy and adenotonsillectomy. STUDY DESIGN/SUBJECTS: Cross-sectional survey. Subjects included all patients from birth to age 29 years who had tonsillectomy or adenotonsillectomy from 1970 to 2005. Study included 8106 patients (median age 8.0 years; range, 6 months to 29 years; male 3646 patients [45%]). Overall tonsillectomy incidence increased from 126 (95% confidence interval [CI], 111-140) per 100,000 person-years in 1970 through 1974 to 153 (95% CI, 139-166) in 2000 through 2005. A dominant factor, adenotonsillectomy incidence rose sharply from 243 (95% CI, 223-261) per 100,000 person-years in 1970 through 1974 to 485 (95% CI, 462-509) in 2000 through 2005. The indication of upper airway obstruction increased from 12 percent of patients in 1970 to 77 percent in 2005. Epidemiological trends in tonsillectomy and adenotonsillectomy have shifted substantially. Overall numbers have increased, and surgical indications have shifted from infection to upper airway obstruction.


[A successful surgical case of atrial septal defect with congenital factor XI deficiency].
We experienced a successful surgical case, who was a 16-year-old boy suffered from atrial septal defect with congenital factor XI deficiency. This disorder is an inherited disorder of blood coagulation characterized by a defect of the intrinsic pathway of thrombin formation and a mild to moderate bleeding tendency, but it can result in severe bleeding in surgical procedures. Open heart surgery in this disorders is rare. Preoperatively we prepared 4000 cc of fresh frozen plasma, but operated without using it. It would appear that the possibility of open heart surgery with congenital factor XI deficiency suggest in this case.


Tuberculosis as an opportunistic disease in persons infected with human immunodeficiency virus.
Tuberculosis, a bacterial disease caused by the Mycobacterium tuberculosis complex, is becoming an increasingly common opportunistic disease in persons infected with the human immunodeficiency virus (HIV). M. tuberculosis is transmitted from person-to-person by airborne droplet nuclei. Persons who are exposed to these droplet nuclei in poorly ventilated environments are at risk of becoming infected with M. tuberculosis. HIV infection is probably the most significant risk factor associated with progression from latent M. tuberculosis infection to active disease. Thus, HIV-infected persons should avoid exposure to M. tuberculosis, they should be screened for evidence of latent infection with the tuberculin skin test, and they should be offered preventive therapy. Because many severely immunosuppressed anergic HIV-infected persons have been found to have an increased risk of developing active tuberculosis, decisions to use preventive therapy should be individualized on the basis of the local prevalence of tuberculosis and drug-resistance patterns. Persons with active tuberculosis should receive at least 6 months of treatment with recommended regimens, preferably with directly observed therapy, to ensure adequate bacteriologic response, completion of therapy, and cure. Chronic suppressive therapy after completion of therapy is currently not recommended.


Benign breast diseases and body mass index: is there a correlation?
Breast cancer is the leading cancer affecting women in America. Body mass index (BMI) is a known risk factor for the development of breast cancer. The relationship of BMI to benign breast disease is less clear. In addition, certain benign pathologies are associated with an increased risk of cancer. We sought to measure the incidence of benign pathologies and to correlate these findings with BMI and age. All patients undergoing breast biopsy at our center from 2000 to 2005 were identified (n = 1717). Age, BMI, family history, sex, and diagnosis were determined. Patients were grouped into BMI, age, and intervention groups. χ(2) (P < 0.05) was used to identify statistical significance. Fibrocystic disease and fibroadenoma were seen with a lower incidence for patients older than 55 years of age, whereas pathologies requiring further surgical intervention were seen in higher proportions in patients older than 55 years of age. All pathologies were noted to decrease with increasing BMI, except for fibroadenoma, which peaked in BMI group 25 to 29.9 kg/m(2). The presence of benign pathologies was associated with age as expected. Interestingly, although BMI is associated with increased risk of breast cancer, increasing BMI was not associated with benign pathologies that are associated with increased risk of breast cancer. Further study of this area is warranted.


Central sleep apnea and associated Chiari malformation in children with syndromic craniosynostosis: treatment and outcome data from a supraregional national craniofacial center.
The association of Chiari malformation Type I (CM-I) with syndromic craniosynostosis (SC) in children is well established. Central sleep apnea (CSA) may subsequently occur. However, sleep studies performed in these patients have been focused mainly on assessing the severity of obstructive sleep apnea. Therefore, the incidence and management of CSA in these patients remains poorly defined. Authors of this study aimed to assess the efficacy of foramen magnum decompression (FMD) in resolving CSA, initially detected incidentally, in a small cohort of patients with CM-I and SC. The clinical data for 5 children who underwent FMD for CSA at Alder Hey Children's Hospital between December 2007 and December 2009 were retrospectively analyzed. Outcomes were evaluated with respect to FMDs by utilizing pre- and postdecompression sleep studies. Of the 5 patients, 2 had Crouzon syndrome and 3 had Pfeiffer syndrome. Patient age at the time of surgery ranged from 1.1 to 12.6 years (median 4.1 years). The median postoperative follow-up was 3.6 years. Sleep studies revealed that 2 children experienced a > 80% reduction in CSAs at 1.5 and 21 months after decompression. The remaining 3 children experienced a > 60% reduction in CSAs when reevaluated between 2 and 10 months after decompression. The associated central apnea index improved for all patients. Findings suggested that FMD is an effective treatment modality for improving CSA in patients with SC and associated CM-I. The use of multimodal polysomnography technology may improve the evaluation and management of these patients.


[Fever and solid tumor: diagnostic value of procalcitonin and C-reactive protein].
Evaluate the clinical utility of procalcitonin (PCT) and C-reactive protein (CRP) as diagnostic of paraneoplastic fever. A prospective analysis of serum levels of PCT and CRP has been conducted on 68 consecutive febrile patients with solid tumour and no neutropenia. The samples were collected at hospital admission. Out of 68 patients, 57 had head and neck cancer. Forty-three patients had signs of infection and 19 had paraneoplastic fever. CRP was not significantly different between the two groups (infected patients: median: 134 mg/L, extremes: 20-569; paraneoplastic fever patients: median: 154 mg/L, extremes: 26-267; P = 0.75 with Mann-Whitney test). On the other hand, PCT was significantly higher in case of infection (median: 0.44 ng/mL, extremes: 0.09-57.4) than in the case of paraneoplastic fever (median: 0.26 ng/mL, extremes: 0.05-1.17; P = 0.01 with Mann-Whitney test). In our study, no paraneoplastic fever patient had PCT level equal or above 2 ng/mL (negative predictive value of 100%).


Degenerative diseases of the vertebral column.
Several distinct degenerative processes affect the articulations of the vertebral column; each is associated with characteristic radiographic and pathologic abnormalities, and many are accompanied by significant clinical manifestations. A discussion of these processes is best accomplished according to the type of joint that is involved. With regard to cartilaginous articulations, of which the intervertebral disk is most important, intervertebral (osteo)chondrosis, spondylosis deformans, and, in the cervical spine, uncovertebral arthrosis are the major degenerative disorders. Osteoarthritis (osteoarthrosis) affects any of the synovium-lined joints of the vertebral column, including the apophyseal, costovertebral, transitional lumbosacral, median atlantoaxial, and sacroiliac articulations. Fibrous articulations, ligaments, or entheses (sites of tendon or ligament attachment to bone) are involved in diffuse idiopathic skeletal hyperostosis, ossification of the posterior spinal ligaments, and Baastrup disease. Of the many complications of these degenerative processes, alignment abnormalities (including segmental instability, degenerative spondylolisthesis, senile kyphosis, and degenerative scoliosis), intervertebral disk displacement, calcification or ossification, and spinal stenosis are the most important.


[The medical angiology outpatient clinic today. The authors' personal experience and operational proposals].
Over the past two decades the demographic characteristics of our population have changed resulting in a major increase in older age groups and a consequent real increase in the prevalence of vascular diseases' of an atherosclerotic type. This had led to an almost exponential increase in the number of requests for instrumental tests, such as Doppler. The aim of this study, the outcome of a review of the patients attending our outpatient clinic over the past twelve months, was to evaluate the current role of a clinic specialising in medical angiology compared to the changing requests for assistance and to elaborate operating proposals for clinics at various levels, aiming to attain the efficacy and efficiency of outpatient activity in accordance with the objective limitations of health spending and current legislation.


Polygalasaponin F inhibits secretion of inflammatory cytokines via NF-κB pathway regulation.
To study the anti-neuroinflammatory mechanisms of polygalasaponin F (PS-F), ELISA method was used to detect the secretion of inflammatory cytokines. Western blot was used to detect the protein expression and phosphorylation levels. Immunofluorescence assay was used to observe the NF-κB nuclear translocation. PS-F could inhibit the release of inflammatory cytokines TNF-α and NO induced by lipopolysaccharides (LPS) and reduce the expression of inducible nitric oxide synthases (iNOS). As for MAPK-signaling pathway, PS-F could only inhibit the phosphorylation levels of p38 MAPK, but did not significantly affect the phosphorylation levels of JNK and ERK1/2 protein kinases. PS-F could inhibit NF-κB nuclear translocation in a dose-dependent manner. The results of Western blot assay were consistent with immunofluorescence assays. Meanwhile, p38-specific inhibitor SB203580 (20 μM) and p65-specific inhibitor PDTC (100 μM) were, respectively, administered as a positive control. In addition, PS-F could significantly inhibit the cytotoxicity of conditioned medium prepared by LPS-stimulated BV-2 microglia (LPS conditioned media) to neuronal PC12 cells and improve cell viability. PS-F inhibits the secretions of neuroinflammatory cytokines by the regulation of NF-κB-signaling pathway.


Antidiuretic hormone and atrial natriuretic peptide during lower body negative or positive pressure in hypertensive patients with and without left ventricular hypertrophy.
Aim of the study was to evaluate the effect of cardiopulmonary receptors activation and deactivation on antidiuretic hormone (ADH) and atrial natriuretic peptide (ANP) incretion in hypertensive and normotensive subjects. Twenty-one male subjects, 7 normotensives and 14 mild hypertensives, 7 without and 7 with left ventricular hypertrophy (LVH) were admitted to the study. Each subject underwent selective loading and unloading of cardiopulmonary receptors, by application of a positive (LBPP) or negative (LBNP) pressure to the lower body. Blood samples were taken for measurement of ANP, ADH, PRA, immunoreactive renin, aldosterone, noradrenaline and adrenaline. ADH plasma concentration increased during cardiopulmonary receptors inhibition, but this increase became statistically significant (p less than 0.05) at a step of LBNP (-40 mm Hg), in which an involvement of the sinoaortic receptors cannot be excluded. ANP plasma levels increased progressively during LBPP (p less than 0.05 at least). These changes were significantly reduced in hypertensive patients with LVH.


Utilizing bimolecular fluorescence complementation (BiFC) to assay protein-protein interaction in plants.
Protein function is often mediated by the formation of stable or transient complexes. Here we present a method for testing protein-protein interactions in plants designated bimolecular fluorescence complementation (BiFC). The advantages of BiFC are its simplicity, reliability, and the ability to observe protein-protein interactions in different cellular compartments including membranes. BiFC is based on splitting the yellow fluorescent protein (YFP) into two nonoverlapping N-terminal (YN) and C-terminal (YC) fragments. Each fragment is cloned in-frame with a gene of interest, enabling expression of a fusion protein. Reconstitution of the fluorescing YFP chromophore takes place upon interaction of protein pairs that are coexpressed in the same cells.


Variable extension of the transitional zone in the medial structure of carotid artery tripod.
The goal of our study was to demonstrate the extension of the transitional zone (TZ) between elastic and muscular medial structure in carotid artery tripod. A histologic study of 56 probes from 8 carotid artery tripods was performed. The probes were obtained from autopsies of 4 adults (mean age: 47.5 years, range 38 to 55 years) and were taken from 7 different topographic sites. At each level of the CCA (at 1 cm and 2 cm proximal to the bifurcation as well as at the bifurcation) we observed an elastic arterial type in 24 (42.8%) probes with 11 to 20 (medium 15.0) elastic fibers per view field (200 x magnification) in the media. In contrast the histologic structure of the ICA and ECA varied as follows: in 8 sections (14.3%) elastic arterial type with 11 to 16 (medium 13.1) elastic fibers, in 11 sections (19.6%) muscular arterial type with 2 to 5 (medium 3.5) elastic fibers and in 9 sections (16.1%) a transitional arterial type with 6 to 8 (medium 6.7) elastic fibers in the media. Atherosclerotic lesions have prevented the assessment of the arterial type in 4 probes (7.1%). The TZ in the medial structure of carotid artery tripods is exclusively localized in the ICA/ECA but not in the CCA. The ICA/ECA presented a TZ with a length up to 0.5 cm (4 probes; 25%), up to 1.5 cm (4 probes; 25%) and longer than 1.5 cm (6 probes; 37.5%). In this study we confirmed that in the carotid artery tripod, a TZ--an arterial segment with transition from elastic to muscular type--does exist, involving a variable length. Furthermore studies on the impact of the biomechanical properties of the TZ as a potential factor in atherosclerotic disease are justified. In addition, the complex biomechanical behavior of the TZ should be considered prior to interventional procedures.


Association of Toll-like receptor 4 single-nucleotide polymorphisms Asp299Gly and Thr399Ile with the risk of primary open angle glaucoma.
Toll-like receptor 4 (TLR4) is a transmembrane receptor that mediates immune responses to exogenous and endogenous ligands. Previously, non-coding single nucleotide polymorphisms (SNPs) in the TLR4 gene were related to primary open angle glaucoma (POAG). This study was undertaken to investigate whether coding TLR4 Asp299Gly (rs4986790 A/G) and Thr399Ile (rs4986791 C/T) are associated with POAG in a Mexican population. One hundred and eighty-seven unrelated Mexican patients with POAG (94 men and 95 women; mean age 66.49 ± 14.3 years) and 109 control subjects (40 men and 69 women; age, 63.28 ± 7.93 years) were included. SNPs Asp299Gly (rs4986790 A/G) and Thr399Ile (rs4986791 C/T) were genotyped by a Taqman® Allelic Discrimination Assayand. Allelic, genotypic, haplotypic, and model-based (dominant, recessive, and codominant) associations of the SNPs with POAG were analyzed using Chi-squared tests or Fisher exact tests and logistic regression. Strong linkage disequilibrium was observed among the SNPs (D' = 0.8692), which were located in one haplotype block. With respect to allelic diversity, the minor allele of both SNPs generates a significantly increased risk of POAG. The minor allele of Asp299Gly conferred the highest increased risk of POAG (P = 0.0054, OR = 4.47, 95% CI = 1.46-13.70). With regard to genotypic diversity, individuals carrying the minor allele of Asp299Gly and Thr399Ile had a significant increased risk for POAG with OR of 4.47 (P = 0.054, 95% CI = 1.30-15.35) and 3.5, respectively (P = 0.012, 95% CI = 1.17-10.44). Haplotype analysis was non-significant. TLR4 coding SNPs Asp299Gly and Thr399Ile might be used as genetic susceptibility alleles for POAG in Mexican population. Our findings support the role of TLR4 in the pathophysiology of glaucoma.


Herbal Medicine, Oreongsan for Recurrent Chronic Subdural Hematoma: A Case Report.
Recently, the East-Asian herbal complex Oreongsan (Goreisan in Japanese and Wulingsan in Chinese) has been noted for its usefulness in preventing postoperative recurrence of chronic subdural hematoma (CSDH). Here, we present a case of CSDH, an 84-year-old man which had recurred four times, despite three previous burr-hole surgeries. Following the fourth burr-hole surgery, an herbal medicine called Oreongsan (TJ-17, Tsumura, Japan, 15g/d) was administered to the patient in hopes of preventing another CSDH recurrence. This case was successfully treated with Oreongsan in an attempt to prevent any further recurrences. The potential mechanism of action of Oreongsan is hydrostatic modulation. The inhibitory effect of the Oreongsan on aquaporin-4 found on the CSDH membrane could contribute to its ability to reduce and prevent the aggravation of SDH.


Structural stabilization of transthyretin by a new compound, 6-benzoyl-2-hydroxy-1H-benzo[de]isoquinoline-1,3(2H)-dione.
Familial amyloid polyneuropathy (FAP) is a genetic, adult-onset, neurodegenerative disorder caused by amyloid formation of transthyretin (TTR), a thyroxine-binding protein. Mutation in TTR causes a propensity of TTR tetramer to dissociate to monomer, which is the first step to amyloidosis. Thus, a drug that can stabilize the tetramer structure will have therapeutic benefit. Here, by virtual screening and biochemical assays, we identified small molecule 6-benzoyl-2-hydroxy-1H-benzo[de]isoquinoline-1,3(2H)-dione (L6) that can prevent the dissociation of TTR to monomer. X-ray crystallography reveals that L6 binds to the T4 binding pocket of TTR. These findings show that L6 is a candidate TTR stabilizer.


Evaluation of the "Hamilton Thorn computer-based automated system" for dog semen analysis.
An objective evaluation of semen is warranted to assess the canine male fertility and to select appropriate techniques and extenders for semen preservation. With conventional microscopic evaluation, the subjectivity of the analysis makes any comparison of results difficult. In the present study, we validated the Hamilton Thorn computer-aided semen analyzer (HTR-IVOS10 analyzer) for objective assessment of canine semen. A description of fertile canine motility parameters using this analyzer is reported. Semen analysis at 38 degreesC is found to be more optimal and accurate than 30 degreesC. The Makler chamber was preferred to the Cell-vu, which induced a decrease of all semen motility parameters. The repeatability of the measures was good with intra-and inter-assay coefficients of variation below 10% and 20%, respectively, for most of measured parameters. An overestimation of semen concentration, increasing with dilution of semen, was observed when HTR-IVOS10 results were compared with the classical manual Makler cell evaluation. A significant decrease in semen motility parameters was recorded when high semen dilutions were used. Generated from the analysis of 42 mature fertile male beagle dogs, a description of semen motility parameters using the CASA system is presented to serve as reference when comparing Beagle ejaculates both in clinical and experimental studies.


Heme Oxygenase-1 Is Not Decreased in Preeclamptic Placenta and Does Not Negatively Regulate Placental Soluble fms-Like Tyrosine Kinase-1 or Soluble Endoglin Secretion.
Elevated placental release of the antiangiogenic factors, soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sENG), is central to the pathophysiology of preeclampsia. It is widely accepted that heme oxygenase-1 (HO-1) is decreased in preeclamptic placenta and negatively regulates sFlt-1 and sENG production. We set out to verify these contentions. There was no difference in HO-1 mRNA or protein levels in preterm preeclamptic placentas (n=17) compared with gestationally matched controls (n=27). In silico analysis of microarray studies did not identify decreased placental HO-1 expression in preeclamptic placenta. Silencing HO-1 in primary trophoblasts did not affect sFlt-1 protein secretion after 24 or 48 hours. Silencing nuclear factor (erythroid-derived 2)-like 2 (transcription factor that upregulates HO-1) in trophoblasts also did not affect sFlt-1 secretion. Administering tin protoporphyrin IX dichloride (HO-1 inhibitor) or cobalt protoporphyrin (HO-1 inducer) into placental explants did not affect sFlt-1 or sENG secretion. Silencing HO-1 in 2 types of primary endothelial cells (human umbilical vein endothelial and uterine microvascular endothelial cells) significantly increased sFlt-1 secretion but not sENG secretion. However, HO-1 silencing selectively increased mRNA expression of sFlt-1 i13 (generically expressed sFlt-1 variant) but not of sFlt-1 e15a (sFlt-1 variant mainly expressed in placenta). Furthermore, adding tin protoporphyrin IX dichloride decreased sFlt-1, whereas adding HO-1 inducers (cobalt protoporphyrin, dimethyl fumarate, and rosiglitazone) either had no effect or increased sFlt-1 or sENG secretion (these trends are opposite to what is expected). We conclude that HO-1 expression is not decreased in preeclamptic placenta and HO-1 does not negatively regulate placental sFlt-1 and sENG secretion in placental or endothelial cells.


Blood pressure response to intravenous noradrenaline is influenced by substance P in normotensive and hypertensive rabbits.
Substance P (SP) applied to the cerebral ventricles elevates arterial blood pressure (BP). This elevation is mediated by an increased sympathetic tone and, moreover, could be attenuated by intravenous (i.v.) SP. To check the assumption that a change of adrenoceptor efficiency might contribute to the attenuation, SP (5 ng/kg per min) was continuously given i.v. and 20 micrograms SP injected intracerebroventricularly (i.c.v.) in anaesthetized normotensive as well as hypertensive rabbits. The BP response to 5 micrograms noradrenaline (NA) was checked within a control period before SP was given and rechecked 30 min after i.c.v. SP with continued i.v. infusion. Under these circumstances SP altered the BP response to NA qualitatively and quantitatively. An initial BP drop was followed by a rise which was remarkably smaller in magnitude than that obtained within the control period. The BP response to phenylephrine (PHE), however, was not altered by the identical SP protocol. Substance P application only i.v. or only i.c.v. was ineffective. Moreover, no differences could be revealed between normotensive and hypertensive animals in the BP response to NA. It is concluded that the beta-adrenoceptor efficiency changed similarly in the normotensive and hypertensive rabbits with i.v. SP, provided sympathetic tone had previously increased. In this manner i.v. SP might modify an exaggerated sympathetic tone which might be implicated in the pathogenesis of arterial hypertension.


L-type Ca(2+) channel regulation by pituitary adenylate cyclase-activating polypeptide in vascular myocytes from spontaneously hypertensive rats.
Pituitary adenylate cyclase-activating polypeptide (PACAP), a vasoactive peptide, modulates the L-type Ca(2+) channel current (L channel current) in vascular smooth muscle cells (VSMC) through activation and integration of two intracellular pathways, protein kinase A and protein kinase C (PKC). In the present study we compared the effects of PACAP on the L channel current in VSMC from the spontaneously hypertensive rats (SHR) and normotensive controls, Wistar Kyoto rats (WKY). We found that compared with WKY, VSMC from SHR had a higher L channel current density. Stimulation by PACAP (10 nM) caused an increase in the amplitude of the whole cell current and prolonged open time in VSMC from SHR and WKY, with the increase greater in SHR. These effects of PACAP on the L channel current was mimicked by an activator of PKC. In contrast, PACAP caused a smaller increase in cAMP accumulation in VSMC from SHR than WKY, and there was no difference in the inhibitory effect of 8-bromo-cAMP on the L channel current from both type of cells. The greater increase in amplitude of the L channel current by PACAP in VSMC from SHR persisted in the presence of adenosine cyclic 3',5'-monophosphothioate, Rp-isomer, a cAMP antagonist, but not calphostin C, a PKC inhibitor. Taken together, our results show an increase in L channel current density and an enhanced PACAP effect on the L channel current in VSMC from SHR compared with WKY. This difference in PACAP response appears to be predominately secondary to an increased PKC sensitivity.


Clinicopathologic implication of hepatic progenitor cell marker expression in hepatoblastoma.
Hepatic progenitor cells (HPCs) are thought to play a role in hepatoblastoma, as hepatoblastomas are characterized by an immature histology and a wide variety of cell lineages. We aimed to investigate the extent of expression of HPCs marker and its clinical implication in hepatoblastoma. We collected 61 hepatoblastomas and 9 childhood hepatocellular carcinomas (HCCs) and performed immunohistochemistry for HPC markers, including cytokeratin 19 (CK19), octamer-binding transcription factor 3/4 (Oct-3/4), epithelial cell adhesion molecule (EpCAM), and delta-like 1 homolog (DLK1). Of the hepatoblastoma samples, 27/61 (44.3%), 21/61 (34.4%), 51/61 (83.6%) and 56/61 (91.8%) exhibited positivity for CK19, Oct-3/4, EpCAM and DLK-1, respectively. For HCCs, the rates of expression were 22.2% (CK19), 77.8% (EpCAM) and 77.8% (DLK-1). Oct-3/4 was not expressed in HCC cells. Hepatoblastomas with a poorly differentiated epithelial component had a higher incidence of CK19 and Oct-3/4 expression than those with a well differentiated epithelial component (p=0.005 and 0.037, respectively). Higher disease stage of hepatoblastoma was correlated with CK19 expression (p=0.043). Oct-3/4-positive hepatoblastomas were associated with short disease-free survival (p=0.035). Both hepatoblastomas and childhood HCCs, therefore, exhibit characteristics of HPCs, and the poor prognosis of patients with Oct-3/4-positive hepatoblastoma suggests that stem-like properties affect hepatoblastoma pathogenicity.


Intraocular Pressure After Implantation of a Phakic Collamer Intraocular Lens With a Central Hole.
To investigate the middle-term intraocular pressure (IOP) results after implantation of a posterior chamber collagen copolymer phakic intraocular lens (IOL) (V4c Visian; STAAR Surgical Co., Nidau, Switzerland) with a central hole in patients with myopia. This retrospective study enrolled patients who had implantation of a phakic IOL with a central hole. IOP, central vault, and adverse events were evaluated 1, 3, 6, 12, and 24 months postoperatively. The study enrolled 763 eyes (384 patients, 128 men and 256 women). Mean follow-up was 7.0 ± 7.2 months (range: 1 to 24 months). The mean IOP was 13.2 ± 2.1 mm Hg preoperatively. Postoperatively, the mean IOP was 12.4 ± 1.7 mm Hg at 1 month, 12.5 ± 1.8 mm Hg at 3 months, 12.6 ± 1.3 mm Hg at 6 months, 12.6 ± 1.4 mm Hg at 12 months, and 12.7 ± 1.1 mm Hg at 24 months. Only one case (0.13%) presented an increased IOP (> 21 mm Hg) during the observation period. No pupillary block or acute angle closure was recorded. IOP at the final follow-up visit was 12.8 ± 1.3 mm Hg. Implantation of central hole phakic IOL in myopic patients provided good and safe IOP outcomes throughout the 24-month observation period. [J Refract Surg. 2017;33(4):244-249.].


In vitro effects of diacetoxyscirpenol (DAS) on human and rat granulo-monocytic progenitors.
Diacetoxyscirpenol (DAS) is a trichothecene mycotoxin produced by various species of fungi. Trichothecenes are known as major contaminants of cereals and cereal-containing foods. DAS has been detected in agricultural products worldwide and persists in products after processing. In human as well as in animals, DAS consumption has been shown to induce haematological disorders (neutropenia, aplastic anemia). Granulo-monocytic progenitors (CFU-GM) from human umbilical cord blood and rat bone marrow have been cultured in the presence of DAS (from 10(-8) M to 5 x 10(-10) M) for 14 days. Study of concentration and effect relationships have shown a sharp effect of DAS on rat CFU-GM between 10(-7) M and 10(-8) M, while human CFU-GM are able to grow in the presence of 10(-8) M of the toxin. IC50 values on day 14 are respectively, 7.6 x 10(-9) M for human CFU-GM and 6.2 x 10(-9) M for rat CFU-GM.


Propagation of guided elastic waves in 2D phononic crystals.
The phononic band structure of two-dimensional phononic guides is numerically studied. A plane wave expansion method is used to calculate the dispersion relations of guided elastic waves in these periodic media, including 2D phononic plates and thin layered periodic arrangements. We show that, for any guided elastic wave, Lamb or generalised Lamb modes, stop bands appear in the dispersion curves, displaying a phononic band structure in both cases.


Clinical features of vascular insufficiency in the limbs.
Drugs (nicotine, beta blockers) can exacerbate symptoms of vascular insufficiency. Vasodilators and anticoagulants have no place in the treatment of chronic insufficiency. Analgesics rarely, if ever, relieve ischaemic pain at rest. Ischaemic pain at rest and gangrene are absolute indications for surgical restoration of the circulation. Spasm is never the determining factor in acute insufficiency associated with trauma. Surgical restoration of circulation is essential in most cases of acute insufficiency. The knee joint should be preserved if possible in lower limb amputation, even if proximal vascular reconstruction is necessary. Surgery should not be denied on the basis of age or infirmity. Restoration of normal circulation in the limb is the best treatment for vascular insufficiency. Acute or chronic vascular insufficiency is eminently correctable in most cases. Advances in diagnosis, patient selection, anaesthesia, operative surgery and rehabilitation have rendered reconstructive procedures safe and successful. Early diagnosis and referral to a vascular unit with modern facilities offers a greater chance of salvage of life and limb than that possible in earlier decades. Many patients return to an independent life free of pain and requiring minimal ongoing care.


Glomerular organization in developing olfactory and accessory lobes of American lobsters: stabilization of numbers and increase in size after metamorphosis.
Olfactory glomeruli are columnar and radially arranged at the periphery of the primary chemosensory areas, the olfactory lobes (OLs), in the American lobster Homarus americanus. The number of olfactory glomeruli reaches nearly 100/lobe in midembryonic life, increases rapidly during larval life, and stabilizes at about 200 in juvenile and adult lobsters. The accessory lobes (ALs), higher order integration areas, are composed of cortical columns and of spherical glomeruli. Two populations of spherical glomeruli are defined, the cortical glomeruli located at the bases of the columns, and the medullary glomeruli scattered throughout the ALs. Both cortical columns and spherical glomeruli are seen for the first time in the second larval stage. There are about 1000 cortical columns and 1700 glomeruli/AL in the postlarva and these numbers remain constant during the life of the lobster. In both OLs and ALs, it is the size of the interglomerular spaces and of the glomeruli themselves that increases. Therefore, the data suggest that in both OLs and ALs the glomeruli were already generated when the lobster metamorphoses (stage III to IV) and switches from a planktonic to a benthic existence, and that the new sensory neurons that are formed at each molt in the antennulae grow into existing olfactory glomeruli. Stability of the glomerular population in the primary olfactory centers, once the full complement of glomeruli is acquired, has also been reported in insects, fish, and mammals.


The effect of hypertension and left ventricular hypertrophy on the lower range of coronary autoregulation.
These studies were performed to test the hypothesis that left ventricular hypertrophy arising as a complication of chronic hypertension is associated with impaired coronary autoregulation. Twelve dogs with hypertension and left ventricular hypertrophy (one-kidney, one-clip model) and 11 normal dogs were instrumented and subsequently studied while conscious. Circumflex pressure, measured with an intracoronary catheter, was adjusted to 100, 75, and 40 mm Hg with a hydraulic occluder that was placed proximally. At each circumflex pressure, myocardial perfusion was measured with radioactive microspheres. Reduction of circumflex pressure over this range did not significantly alter heart rate, left atrial pressure, or arterial pressure. In normal dogs, reduction of circumflex pressure did not alter total myocardial perfusion or the transmural distribution of perfusion. In contrast, in dogs with hypertension and left ventricular hypertrophy, circumflex subendocardial perfusion decreased 46% when pressure was decreased from 100 to 40 mm Hg (p less than .05 compared with normal). Autoregulation was quantified for each third of myocardium with the use of autoregulatory gain values (1 = perfect autoregulation; 0 = the absence of autoregulation). For pressure changes of 100 to 75 mmHg, values for autoregulatory gain were near unity for all layers of myocardium in both groups of animals. When pressure was decreased from 75 to 40 mm Hg, values for autoregulatory gain among the normal and hypertensive groups were, respectively: for subepicardium 1 +/- 0.2 (mean +/- SE) vs 0.9 +/- 0.2 (p = NS), for the midwall 0.8 +/- 0.2 vs 0.5 +/- 0.2 (p = NS), and for the subendocardium 0.8 +/- 0.1 vs 0.1 +/- 0.2 (p less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)


Improved measurement of Vub with inclusive semileptonic B decays.
We report a new measurement of the Cabibbo-Kobayashi-Maskawa parameter Vub made with a sample of 9.7 x 10(6) BB- events collected with the CLEO II detector. Using heavy quark theory, we combine the observed yield of leptons from semileptonic B decay in the end-point momentum interval 2.2-2.6 GeV/c with recent CLEO II data on B-->X(s)gamma to find Vub = (4.08+/-0.34+/-0.44+/-0.16+/-0.24)x10(-3), where the first two uncertainties are experimental and the last two are from theory.


Human security: expanding the scope of public health.
Human security is an evolving principle for organizing humanitarian endeavours in the tradition of public health. It places the welfare of people at the core of programmes and policies and is community oriented and preventive. It recognizes the mutual vulnerability of all people and the growing global interdependence that marks the current era. Health is a crucial domain of human security, providing a context within which to build partnerships across disciplines, sectors and agencies. These principles have been demonstrated in field programmes in which healthcare delivery featuring multi-sectoral co-operation across conflict lines has been used to enhance human security. Such programmes can be a model for collaborative action and can create the sustainable community infrastructure that is essential for human security.


Multiple androgen response elements cooperate in androgen regulated activity of the type 1 neutral endopeptidase promoter.
The neutral endopeptidase (NEP) gene is transcriptionally regulated by androgen in prostate cancer cells. We previously identified in the NEP gene an androgen responsive element (NEP-ARE) and an androgen responsive region (NEP-ARR) that together conveyed only moderate androgen-inducibility [Mol. Cell. Endocrinol. 170 (2000) 131]. Therefore, we characterized the entire genomic structure of the NEP gene and identified ARE1 (ACTCAACAttgTGTCCTTT) and ARE2 (CAGGACAtttTGTCCC), which are located in the 3'-untranslated region and in intron 17, respectively. Steroid-dependent enhancement of transcription was assayed by transfecting the pGL-3-luciferase reporter plasmid containing three copies of ARE1 or ARE2 into PC-3 cells. Luciferase activities were increased 3.6-fold (ARE1) and 5-fold (ARE2) by androgen (AR), 4.2-fold (ARE1) and 8.2-fold (ARE2) by dexamethasone, and 3-fold (ARE1) and 4.1-fold (ARE2) by progesterone. Mutation of the ARE1 and ARE2 sequences completely abrogated androgen-inducibility. We next showed that both ARE1 and ARE2 are involved in the transcriptional regulation of the NEP gene, demonstrating in vitro and in vivo binding with AR as determined by electrophoretic mobility gel shift and chromatin immunoprecipitation (ChIP) assays, Furthermore, ARE1 and ARE2 mediate coordinated androgen-inducibility in both an SV40 promoter and the native NEP type 1 promoter. These data indicate the newly identified ARE1 and ARE2 together with the previously identified NEP-ARE function as androgen response elements, and that androgen regulation of the NEP gene is regulated by the coordinated action of multiple AREs in prostate cancer cells.


Mycophenolate mofetil in juvenile dermatomyositis: a case series.
The objective of this study was to report the use of Mycophenolate Mofetil (MMF) in Juvenile Dermatomyositis (JDM). A retrospective chart review of children diagnosed with JDM having received MMF was performed. Response was evaluated 3 months after the onset of MMF by comparing muscle strength and steroid dosage before and after treatment. A good response was defined by global improvement concerning weakness and fatigability as evaluated subjectively by the physician along with a gain of at least 4 points on each of 2 muscle testings (Manual Muscle Testing, MMT and Childhood Myositis Assessment Score, CMAS) and/or a decrease of >15% of the corticosteroid dosage. Eight patients were identified. Except for one, all had received MMF secondary to an initial therapy of conventional immunosuppressants. Six patients showed good response by our predefined criteria. Changes of muscle testing scores ranged between +0 to +21 points (mean = +10.6) for the MMT and between +3 and +11 (mean = +7) for the CMAS. Corticosteroid tapering varied from 0 to 50%, with a mean of 18%. In most cases, follow-up was available for many months (up to 26); overall, we observed only one complication: a transient neutropenia in a patient concurrently receiving another immunosuppressant. This small series is the first published report on the use of MMF in JDM and suggests it is safe. Prospective larger studies are required to further elucidate the use of MMF in JDM.


Sulfate content and specific glycosaminoglycan backbone of perlecan are critical for perlecan's enhancement of islet amyloid polypeptide (amylin) fibril formation.
Islet amyloidosis is characterized by the deposition and accumulation of amylin in pancreatic beta-cells and is observed in 90% of patients with type 2 diabetes. Previous studies have also revealed the presence of the specific heparan sulfate proteoglycan, perlecan, colocalized to islet amyloid deposits, similar to perlecan's known involvement with other amyloid proteins. In the present study, perlecan purified from the Engelbreth-Holm-Swarm (EHS) tumor was used to define perlecan's interactions with amylin (i.e., islet amyloid polypeptide) and its effects on amylin fibril formation. Using a solid phase-binding immunoassay, human amylin, but not rat amylin, bound immobilized EHS perlecan with a single dissociation constant (Kd) = 2.75 x 10(-6) mol/l. The binding of human amylin to perlecan was similarly observed using perlecan heparan sulfate glycosaminoglycans (GAGs), and was completely abolished by 10 micromol/l heparin. Using thioflavin T fluorometry, Congo red staining, and electron microscopy methodology, intact perlecan was found to enhance amylin fibril formation in a dosage-dependent manner, with the majority of these effects attributed to the heparan sulfate GAG chains of perlecan. Other sulfated GAGs and related macromolecules were also effective in the enhancement of amylin fibril formation in the order of heparin > heparan sulfate > chondroitin-4-sulfate = dermatan sulfate = dextran sulfate > pentosan polysulfate, implicating the importance of the specific GAG/carbohydrate backbone. The sulfate content of heparin/heparan sulfate was also important for the enhancement of amylin fibril formation in the order of heparin > N-desulfated N-acetylated heparin > completely desulfated N-sulfated heparin > completely desulfated N-acetylated heparin. These studies suggest that the enhancement effects of perlecan on amylin fibril formation are mediated primarily by both specific GAG chain backbone and GAG sulfate content, and implicate perlecan as an important macromolecule that is likely involved in the pathogenesis of islet amyloidosis.


Role of liposomal anthracyclines in the treatment of multiple myeloma.
Patients with multiple myeloma (MM) typically respond to initial chemotherapy, but almost all patients relapse with a median survival of approximately 5 years. Combining vincristine and conventional doxorubicin with oral dexamethasone (VAD) or reduced-dose dexamethasone (VAd) provides rapid response in many patients, but its use is limited by toxicity concerns and the inconvenience of continuous infusions in each cycle. Use of pegylated liposomal doxorubicin (Doxil/Caelyx [PLD]) and vincristine intravenous injection with oral dexamethasone (DVD) or reduced-dose dexamethasone (DVd) is safe and effective for the management of newly diagnosed or relapsed/refractory MM. Controlled trials showed that DVD/DVd is at least as effective as VAD/VAd for the treatment of MM, but DVd is associated with less neutropenia and alopecia in addition to requiring fewer days in the hospital or clinic for drug administration. DVd therapy has also been reported to be associated with an antiangiogenic effect not observed with VAD. Another liposomal anthracycline, liposomal daunorubicin (DaunoXome [DNX]), has been investigated in MM and preliminary data suggest that it is safe and effective, but studies comparing it with other regimens have not been reported. Early results from ongoing trials suggest that adding thalidomide, bortezomib, or other immune modulators to PLD-based chemotherapy may improve efficacy.


Identification of key gene modules and pathways of human breast cancer by co-expression analysis.
Breast cancer is the most common and aggressive tumor causing injury to women world wide. Although gene expression analysis had been performed previously, systemic co-expression analysis for this cancer is still lacking to date. We attempted to identify the critical modules of breast cancer. Co-expression modules were established with the help of WGCNA and the interactions among them were performed by R language. Biological process and pathways analysis of co-expression genes were figured out by GO and KEGG functional enrichment analysis using DAVID dataset. In this study, expression data of 4,000 genes from 136 samples with breast cancer was used for the establishment of co-expression modules. And nine modules were identified. There was much higher scale independence among different modules by interactions analysis. Moreover, there was an obvious difference in adjacency degree among different modules. The most enriched pathways as immune response and ubiquitin-mediated proteolysis were identified as the most critical modules of breast cancer by GO and KEGG enrichment analysis. Our result demonstrated that immune response and ubiquitin-mediated proteolysis could serve as prognostic and predictive markers for the occurrence of breast cancer, providing evidence for further analysis in the prognosis and treatment of breast cancer.


CLOCK gene 3111C/T polymorphism is not associated with seasonal variations in mood and behavior in Korean college students.
The present study tested the potential association between the 3111C/T polymorphism of the CLOCK gene and seasonal variations in mood and behavior. A total of 297 Korean college students were genotyped for the CLOCK polymorphism and the seasonal variation was evaluated using the Seasonal Pattern Assessment Questionnaire (SPAQ). The seasonality scores were not different between CLOCK gene variants (P > 0.05). Comparison between seasonals (syndromal plus subsyndromal seasonal affective disorder according to SPAQ) and non-seasonals found no significant difference in frequencies of genotypes (P > 0.05). These findings suggest that the CLOCK polymorphism does not play a major role in susceptibility to seasonal variations in a Korean population.


Enantioselective recognition between chiral alpha-hydroxy-carboxylates and macrocyclic heptadentate lanthanide(III) chelates.
Three novel heptacoordinated Ln(III) complexes (Ln = Gd and Yb) have been synthesized and investigated by (1)H NMR spectroscopy. These complexes contain two stereogenic centers, one associated with a deltadeltadeltadelta or lambdalambdalambdalambda conformation of the ethylenediamine moieties in the tetraazamacrocycle and the latter arises from the orientation (Delta or Lambda) of the coordinating arms. Evidence has been gained for the occurrence of a fast exchange between all the possible conformers. Upon addition of several (S)-alpha-hydroxy-carboxylate substrates, the formation of stable ternary adducts has been obtained. Their (1)H NMR spectra are consistent with the presence of two diastereoisomers differing in the conformation adopted by the macrocyclic ligand wrapping the lanthanide(III) ion. The interaction leading to the formation of the ternary complexes is enantioselective depending on the hydrophilicity of the alpha-hydroxy-carboxylate.


Diagnosing Helicobacter pylori in vivo by confocal laser endoscopy.
Confocal laser endomicroscopy enables subsurface microscopic imaging of living tissue during ongoing endoscopy. This case report describes the in vivo detection of Helicobacter pylori by endomicroscopy. Endomicroscopy (Pentax, Tokyo, EC-3870CIFK) was performed by using two different contrast stains: Topical Acriflavine in addition to intravenously applied fluorescein netted the surface and allowed identification of focal accumulation of Helicobacter pylori at the surface and in deeper layer of the gastric epithelium. Biopsies were performed at the antrum and corpus for urease testing and histology. In addition, biopsies were cultured for Helicobacter pylori. Cultured bacteria were re-assessed ex vivo using confocal microscopy with and without acriflavine staining. Helicobacter pylori infection could be detected in a 70-year-old male by endomicroscopy. Accumulated, as well as single bacteria, could be observed and the distinct shape and flagella of Helicobacter pylori could be identified. Helicobacter pylori infection was proved by histology. Furthermore, ex vivo examination of cultures proved the presence of Helicobacter pylori and the active uptake of acriflavine into the bacteria. Endomicroscopy is a new diagnostic approach, which enables the immediate diagnosis of Helicobacter pylori in vivo during standard video endoscopy.


Simple structural modifications confer cytotoxicity to allobetulin.
A variety of allobetulin derivatives was synthesized from allobetulin or allobetulone. These compounds were screened for their cytotoxic activity using a photometric SRB assay employing six different human tumor cell lines. In summary, opening of ring A of allobetulin in general lowers the cytotoxicity, but the 2,3-seco diethyl ester was highly cytotoxic and remarkable selective for A549 lung carcinoma cells while being significantly less cytotoxic for non-malignant mouse fibroblasts. The introduction of an amino group at position C-3 in the allobetulin skeleton enhances cytotoxicity and furnishes highly cytotoxic compounds. Their selectivity to distinguish between cancer cell and non-malignant cell depends on the configuration at position C-3.


Unsuspected FDG-PET findings in the follow-up of patients with lymphoma.
18F-Fluorodeoxyglucose-positron emission tomography (FDG-PET) plays an increasing role in the management of patients with lymphoma, for which it is successfully used for staging and treatment monitoring. We report seven patients with a history of lymphoma who presented a positive FDG-PET suggestive of lymphoma relapse and for which FDG-PET oriented biopsies revealed alternative diagnoses. Early in lymphoma follow-up, persistence of focal increased FDG activity corresponded to inflammatory or infectious lesions in two patients: one aspergillosis and one sarcoidosis. Later in the follow-up, five cases of secondary malignancies were identified (three lung cancers, one epidermoid carcinoma, and one villous tumor) in this particularly exposed population. The routine use of FDG PET to evaluate lymphoma significantly increases the probability of detecting unexpected diseases. These cases illustrate the potential pitfalls in PET follow-up. Because FDG is not lymphoma-specific, a relapse suspected only on FDG-PET imaging requires biopsy, as alternative diagnoses--infectious or malignant--are possible. Our data draws clinician's attention to potential false-positive FDG-PET findings, which may lead to therapeutic mistakes. Our data also suggests that FDG-PET might be a new imaging modality for long-term monitoring of late effects, especially second cancer occurrence.


Detailed esophageal function and morphological analysis shows high prevalence of gastroesophageal reflux disease and Barrett's esophagus in patients with cervical inlet patch.
Although the pathogenesis of cervical inlet patch (CIP) is not fully understood, most authors consider it as a congenital abnormality, whereas others surmise it to be related to gastroesophageal reflux disease (GERD). We aimed to evaluate esophageal function and the prevalence of GERD and Barrett's esophagus in patients with CIP. GERD is defined by the presence of erosive esophagitis or an abnormal pH monitoring. Seventy-one consecutive patients with endoscopic and histological evidence of CIP were prospectively evaluated. Esophageal symptom analysis, 24-hour simultaneous biliary reflux and double-channel pH-monitoring, and esophageal manometry were carried out in 65/71 (92%) patients and in 25 matched controls. Six patients were not suitable for testing and were, therefore, excluded. The histological evaluation of the heterotopic islands showed cardia and/or oxyntic mucosa in 64/65 (98%) patients and specialized intestinal metaplasia (SIM) in one patient (2%). The cardia and/or oxyntic mucosa was accompanied by focally appearing pancreatic acinar metaplasia and pancreatic ductal metaplasia in 7/64 (11%) and in 1/64 (2%), superficial mucous glands in 6/64 (9%), and SIM in 2/64 (3%) cases. In total, SIM was present in three patients (5%), and one of them had low-grade dysplasia. At the gastroesophageal junction, 28 (43%) patients had columnar metaplasia, including nine (14%) patients with SIM. Erosive esophagitis was present in 37 (57%) cases. Thirty-two patients (49%) had abnormal acid reflux in the distal and 25 (38%) in the proximal esophagus. Abnormal biliary reflux was present in 25 (38%) cases. On the basis of endoscopic and pH studies, GERD was established in 44/65 (68%) patients. Typical reflux symptoms were common (33/65, 51%). The combined 24-hour biliary and double-channel pH-monitoring detected significantly more significant acidic reflux at both measurement points and significantly longer bile exposure time in the distal esophagus in patients with CIP. Acid secretion in the CIP was detected in three (5%) cases. Esophageal manometry revealed decreased LES pressure and prolonged relaxation with decreased peristaltic wave amplitude, and an increased number of simultaneous contractions in the esophageal body. The detailed evaluation of the esophageal morphology and function in subjects with CIP showed a high prevalence of GERD and Barrett's esophagus. Further studies are needed to evaluate whether combined acidic and biliary reflux is able to promote similar histomorphological changes in the CIP, as it is shown distally in patients with Barrett's esophagus.


Comparison of morning vs bedtime administration of the combination of valsartan/amlodipine on nocturnal brachial and central blood pressure in patients with hypertension.
The aim of this study was to compare the effect of morning and bedtime administration of valsartan/amlodipine combination therapy (80/5 mg) on nocturnal brachial and central blood pressure (BP) measured by ambulatory BP monitoring in patients with hypertension. This was a 16-week prospective, multicenter, randomized, open-label, crossover, noninferiority clinical trial. Patients underwent 24-hour ambulatory BP monitoring at randomization, at switching, and at the end of the study. Twenty-three patients (mean age, 68.0 years) were studied. The difference in nocturnal brachial systolic BP between the morning and bedtime administrations of combination valsartan/amlodipine was -3.2 mm Hg, and the two-sided 95% confidence interval ranged from -6.8 to 0.4 mm Hg. The difference in nocturnal central systolic BP was -4.0 mm Hg (95% confidence interval, -7.6 to -0.4 mm Hg). The upper limit of the 95% confidence interval was below the margin of 3.0 mm Hg in both nocturnal brachial and central systolic BP, confirming the noninferiority of morning administration to the bedtime administration of valsartan/amlodipine combination therapy.


2H NMR study of 2D melting and dynamic behaviour of CDCl3 confined in ACF nanospace.
Two-dimensional melting of trichloromethane (chloroform) confined in activated carbon fibre was investigated using differential thermal analysis and (2)H NMR techniques. Differential thermal analysis revealed a thermal anomaly with an endothermic peak at 269 K, which was distributed from 250 K to 287 K on the heating direction. This anomaly was also observed upon cooling at the same temperature. Furthermore, (2)H NMR revealed that slow motion such as molecular hopping and/or diffusion of CDCl(3) in ACF affected the spectral line width. The temperature dependence (Arrhenius plot) of the spectral line width showed an inflection point at 227 K. The activation energy of molecular motion of CDCl(3) in ACF was 4 kJ mol(-1) at temperatures greater than 227 K and 7.7 kJ mol(-1) at temperatures less than 227 K. Reduction of the activation energy suggests that the average intermolecular distance between CDCl(3) molecules enlarges above the inflection point. The difference of activation energy (3.7 kJ mol(-1)) is close to the enthalpy of fusion in typical plastic crystals. These results reveal that the thermal anomaly and the transition of dynamic process correspond respectively to melting of CHCl(3) in ACF and the pre-melting phenomenon.


Rapid diagnosis of congenital adrenal hyperplasia by high performance liquid chromatography.
This manuscript describes methods for the quantitation of serum concentrations of 17-hydroxyprogesterone and 11-deoxycortisol (Compound S) employing reverse-phase high-performance liquid chromatography (HPLC) and ultraviolet (UV) absorbance which are applicable to the diagnosis of congenital adrenal hyperplasia due to deficiencies of 21- and 11-hydroxylase activities, respectively. These methods are simple, specific, precise and rapid. Data obtained by the HPLC-UV methods are highly correlated (p less than 0.001) with radioimmunoassay measurements.


Sources of interobserver variation in histopathological grading of cervical dysplasia.
The present study aimed to assess where the interobserver variation in grading cervical dysplasia stems from. Four experienced pathologists examined 93 histological slides, after they agreed on which morphological characteristics should be considered relevant for grading. They scored 6 morphological characteristics for each slide and assigned it to a degree of dysplasia. Compared to a previous study, the interobserver variation showed a statistically significant improvement: the weighted group kappa value increased from 0.55 to 0.69. For the scores of the individual characteristics considerable interobserver variation was observed: weighted group kappa values ranged from 0.28 to 0.49. The pathologists slightly differed in which characteristics they considered most important for their grading. The agreement on the degree of dysplasia turned out to be better than the agreement on the morphological characteristics on which this diagnosis is based. In the discussion, a few explanations for this paradoxical finding are put forward.


Blood oxidant/antioxidant status of atherosclerotic patients.
In the present study, it is aimed to investigate oxidant/antioxidant status of plasma and erythrocytes from atherosclerotic patients and to establish the possible role of oxidant stress in the formation and progression of atherosclerosis. Antioxidant potential (AOP) values and malondialdehyde (MDA) levels were studied in erythrocyte and plasma samples from 40 atherosclerotic patients and 38 healthy controls. A total of 13 subjects in each group were smokers. AOP was found unchanged in erythrocytes but lower in plasma samples (P<0.0005) from atherosclerotic patients as compared with those of the controls. MDA levels were however higher in erythrocyte hemolysate (P<0.025), erythrocyte membrane (P<0.0005) and blood plasma samples (P<0.0005) from atherosclerotic patients than those of the controls. Moreover, AOP was found to be lower in plasma samples of smoker patients than that of non-smoker patients (P<0.05). In the control group, erythrocyte MDA level was higher in smoker group than that of non-smoker group (P<0.05). Results reveal the presence of oxidant stress in the blood samples from patients with atherosclerosis. It seems antioxidant therapy might give beneficial results for atherosclerotic patients.


Clinical Impact of Flat Panel Volume CT Angiography in Evaluating the Accurate Intraoperative Deployment of Flow-Diverter Stents.
The deployment of flow-diverter stents may be difficult to analyse on regular DSA. The purpose of our study was to investigate the clinical impact of stent-dedicated flat panel volume CT angiography to evaluate intraoperatively the satisfactory deployment of flow-diverter stents. From January 2009 to April 2015, 83 consecutive patients (mean age, 51 years; 62 women) were treated in our institution with flow-diverter stents. Eighty-seven aneurysms (82 unruptured, 5 ruptured; 77 anterior, 10 posterior circulation) were treated in these 83 patients (4 patients had 2 aneurysms, both treated by means of flow-diverter stents). One patient was treated for a traumatic carotid cavernous fistula. In 80% of the cases (68/85) a flat panel volume CT angiography was performed in the angiographic suite just after the flow-diverter stent deployment. Stent visualization was assessed by 2 independent reviewers. The clinical impact of stent malapposition was evaluated. Flow-diverter stent visualization was satisfactory in 73.5% of the cases. In 2 cases (2.9%) the flat panel volume CT angiography prompted the operator to perform an additional intrastent angioplasty for a condition that was previously underestimated. Four patients (4.7%) experienced acute thromboembolic complications; 3 others had delayed thromboembolic complications. Only 1 of these patients had thromboembolic complications (acute or delayed) related to stent misdeployment, which was easily managed intraoperatively with no clinical consequence. Flat panel volume CT angiography is an interesting tool to depict flow-diverter stent misdeployment and may encourage the operator to perform intrastent angioplasty (2.9% of the cases in our experience) to reduce the risks of thromboembolic complications.


The value of microbiological screening in cleft lip and palate surgery.
This study was performed to investigate whether nasal and oropharyngeal microbiological swabs taken prior to cleft lip and palate surgery correlated with the oronasal flora at the time of surgery and whether specific culture results affected surgical outcome. Prospective audit set in two designated U.K. cleft centers each with a single surgeon. Nasal and oropharyngeal microbiological swabs were taken within 2 weeks prior to surgery and again on the operating table. Adverse outcome measures included postoperative pyrexia, wound dehiscence, or fistula formation. One hundred forty-four cases were recruited over 12 months. Nasal swabs cultured organisms significantly more often than oropharyngeal swabs (p < .0001). No significant difference was detected in the number of cases with a positive microbiology culture preoperatively compared with perioperative sampling (48% and 50%). The specific organisms cultured from preoperative swabs were the same as those cultured at surgery in only half of cases. Preoperative microbiology swabs were poorly predictive of the oronasal flora at surgery. Antibiotic treatment of patients with positive preoperative microbiology did not significantly reduce the incidence of bacterial colonization or significantly alter clinical outcome. Preoperative microbiological investigation is not helpful in predicting the nasal and oropharyngeal flora at the time of surgery. Further, culture results did not correlate with postoperative outcome, regardless of whether pre- or perioperative antibiotic therapy was instigated. This evidence suggests that microbiology screening swabs are an unnecessary investigation.


Selective ionization of oriented nonpolar molecules with asymmetric structure by phase-controlled two-color laser fields.
We report on the selective ionization of oriented nonpolar molecules with asymmetric structure by using phase-controlled two-color omega + 2omega laser pulses with an intensity of 1.0 x 10(13) W/cm(2) (tunneling ionization regime) and a pulse duration of 130 fs. The orientation of 1-bromo-2-chloroethane was monitored by the directional asymmetries of the forward-backward emission in dissociative ionization. The observed direction of orientation clearly confirms that molecular orientation is induced not by dynamic orientation but by selective ionization of oriented molecules, which reflects the structure of the highest occupied molecular orbital. This method can be applied for the vast majority of molecules.


Effective modulation of the haematopoietic toxicity associated with zidovudine exposure to murine and human haematopoietic progenitor stem cells in vitro with lithium chloride.
The drug zidovudine (AZT), a synthetic thymidine analogue, has been used in the treatment of acquired immunodeficiency syndrome (AIDS). Clinical use of zidovudine has induced haematopoietic toxicity manifested by anaemia, neutropenia, frequent thrombocytopenia, and overall bone-marrow suppression. The monovalent cation lithium has been shown to be an effective agent capable of modulating several aspects of haematopoiesis such as the induction of neutrophilia, thrombopoiesis, and protection against suppression of haematopoietic progenitor stem cells following exposure to anticancer drugs and/or radiation in the treatment of malignant disease. We here report the results of studies designed to evaluate the effectiveness of lithium in reversing and/or protecting against either murine or human bone marrow derived haematopoietic progenitors, i.e. (CFU-GM, CFU-Meg, and BFU-E) when co-cultured in the presence of zidovudine in vitro. Lithium chloride (LiCl) reversed zidovudine toxicity to either murine or human derived CFU-GM and CFU-Meg that was optimal at a concentration of 1 mM (P less than 0.05). However, the addition of lithium failed to influence zidovudine toxicity toward either murine or human BFU-E. In summary, these results support the scant clinical studies that have described the presence of neutrophilia and/or thrombopoiesis in zidovudine-treated AIDS patients receiving lithium. In addition, these data further confirm the need for more detailed evaluation of lithium as an adjuvant agent to reduce the haematopoietic toxicity associated with the use of antiviral therapy in HIV-infected patients.


Pharmacokinetic interaction between bosentan and the oral contraceptives norethisterone and ethinyl estradiol.
Bosentan has been shown in vitro and in vivo to induce the cytochrome P450 enzymes CYP2C9 and CYP3A4. The present study was conducted to investigate the effect of bosentan on the pharmacokinetics of a combined oral contraceptive. In a randomized, 2-way crossover study, 20 healthy female subjects received Treatments A and B. Treatment A consisted of a single dose of OrthoNovum containing 1 mg norethisterone (norethindrone) and 35 microg ethinyl estradiol. Treatment B consisted of bosentan, 125 mg b.i.d. for 7 days plus concomitant norethisterone and ethinyl estradiol on Day 7. Plasma concentrations of norethisterone and ethinyl estradiol were measured on days of oral contraceptive administration. In the absence of bosentan, the pharmacokinetics of norethisterone and ethinyl estradiol were characterized by Cmax and AUC0-infinity values (95% CI) of 9.8 (8.1, 11.9) ng/ml and 72.9 (57.0, 93.1) ng x h/ml, and 53.0 (47.0, 59.9) pg/ml and 758 (655, 878) pg x h/ml, respectively. Concomitant bosentan did not affect the Cmax but significantly decreased the AUC of norethisterone and ethinyl estradiol by 13.7% (-23.5, -2.6) and 31.0% (-40.5,-20.2), respectively. The maximum decrease in AUC of norethisterone and ethinyl estradiol in an individual subject was 56% and 66%, respectively. Bosentan decreases the AUC of norethisterone and ethinyl estradiol in healthy female subjects. In patients treated with bosentan, reduced efficacy of hormonal contraceptives should be considered.


[Mechanism of trace hyperpolarization of the action potential of snail giant neurons].
Depolarization current decreases and hyperpolarization current increases the amplitude of tracing hyperpolarization of the neuron action potential. Calcium-defficient solution supresses the tracing depolarization, and turns the rhythmical activity of the neuron into the flashlike one. An increase of outer concentration of potassium ions decreases the tracing depolarization. The latter is suppressed completely when the membrane behaves as a potassium electrode. The suppressing effect of the increase of potassium outer concentration on tracing hyperpolarization decreases with a decrease of calcium ions content in the medium. When an active release of sodium ions from the cell is inhibited with DNP and substitution of sodium ions by lithium ions the tracing hyperpolarization of the action potential is suppressed. The tracing hyperpolarization is also suppressed during the shunting of the electrogenic effect of potassium pump with the outcoming current of chlorine ions. It is suggested that the tracing hyperpolarization of the single action potential is due to the calcium-dependent fraction of electrogenic release of sodium ions from the cell.


Proteomic analysis of membrane proteins expressed specifically in pluripotent murine embryonic stem cells.
Embryonic stem cells (ESCs) are established from the inner cell mass of preimplantation embryos, are capable of self-renewal, and exhibit pluripotency. Given these unique properties, ESCs are expected to have therapeutic potential in regenerative medicine and as a powerful tool for in vitro differentiation studies of stem cells. Various growth factors and extracellular matrix components regulate the pluripotency and differentiation of ESC progenies. Thus, the cell surface receptors that bind these regulatory factors are crucial for the precise regulation of stem cells. To identify membrane proteins that are involved in the regulation of pluripotent stem cells, the membrane proteins of murine ESCs cultured with or without leukemia inhibitory factor (LIF) were purified and analyzed by quantitative proteomics. 2-D PAGE-based analysis using fluorescently labeled proteins and shotgun-based analysis with isotope-labeled peptides identified 338 proteins, including transmembrane, membrane-binding, and extracellular proteins, which were expressed specifically in pluripotent or differentiated murine ESCs. Functions of the identified proteins revealed cell adhesion molecules, channels, and receptors, which are expected to play important roles in the maintenance of murine ESC pluripotency. Membrane proteins that are expressed in pluripotent ESCs but not in differentiated cells such as Slc16a1 and Bsg could be useful for the selection of the stem cells in vitro.


In vivo changes in brain catecholamine release from rat hypothalamus following olfactory bulbectomy.
The mechanism eliciting mouse-killing behavior (muricide), induced by bilateral olfactory bulbectomy, has been shown to involve the brain noradrenergic system; this is because muricide is specifically inhibited by the drugs which potentiate the activity of catecholaminergic neurons such as tricyclic antidepressants. Our previous reports also demonstrated that the hypothalamic noradrenaline (NA) contents increased in the rats which exhibited muricide. To further examine the hypothalamic noradrenergic function in muricide, a push-pull perfusion technique was applied for direct measurement of NA release from the lateral (LH) and ventromedial (VMH) hypothalamus in freely moving rats. Subsequently, the perfusates, including catecholamines and their metabolites were measured by means of high-performance liquid chromatography with electrochemical detection (HPLC-ECD). Three days after olfactory bulbectomy, 67% of the rats elicited muricide and NA release from LH tended to decrease. Moreover, 7 days after olfactory bulbectomy, most of the rats elicited muricide and NA release from LH was significantly decreased, but not from VMH. On the other hand, dopamine (DA) release from VMH without LH conversely increased on the 7th day after olfactory bulbectomy. These results suggest that the dysfunction of the noradrenergic system caused by the decrease in NA release from LH played an important role for the incidence of muricide.


Antibody-mediated rejection in ABO compatible husband to wife living donor liver transplant and review of the literature.
Role of donor specific antibodies (DSAs) in liver allograft function has not been fully defined. We report an ABO compatible orthotopic liver transplant case with DSAs to donor HLA, where the patient developed immediate antibody-mediated rejection (AMR).The patient, a 43-year-old female with cirrhosis, underwent ABO-compatible living-donor liver transplant from her husband. On post-operative day (POD)1, serum transaminases were sharply elevated. Retrospective testing of pre-transplant serum demonstrated presence of strong class I and class II anti-HLA antibodies and positive T- and B-cell flow-cytometric crossmatches (FCXM). Transaminase levels improved with plasmapheresis and thymoglobulin. On POD7, her liver enzymes became elevated again and allograft biopsy stained positive for C4d. Patient was treated with intravenous immunoglobulin and rituximab and recovered over time. Pre-transplant sera of patient were retrospectively tested by C1q assay to determine the cytotoxic function of DSAs; DSAs were positive for C1q binding. Our results suggest that pre-liver transplant antibody testing may be helpful in identifying patients at risk for development of AMR.


Malformation of the lumbosacral spinal cord in a case of sacral agenesis.
Agenesis of the sacrum is a rare anomaly that is associated with numerous visceral abnormalities, spinal cord malformation, and lower limb defects. A fatal case of sacral and lower lumbar agenesis in a 3-day-old female infant born at 38 weeks of gestation is reported. The extraneural malformations comprised an imperforate anus, a rectovaginal fistula, and musculoskeletal abnormalities, including several thoracocervical hemivertebrae and aplasia of the sacrum and the fourth and fifth lumbar vertebrae. The cervical and high thoracic spinal cord segments were normal. Disruption of secondary neurulation, possibly due to notochord dysfunction, was suggested by malformation of the ventral half of the lower thoracic spinal cord with relative preservation of the dorsal horns and, more caudally, by loss of all normal histological landmarks, including the central canal. Neither skeletal muscle nor myoblasts were found in muscle compartments that would normally have received motor innervation from the levels of the spinal cord from which anterior horn cells were absent, indicating parallel, segmental failure of myotomal differentiation in the caudal eminence.


Clinical experience with dual action antidepressants in different chronic pain syndromes.
A common psychopharmacology between pain and depression suggests that compounds inhibiting the reuptake of serotonin and/or noradrenaline are likely to produce relief from chronic pain. Indeed tricyclic antidepressants have been a standard treatment of chronic pain for many years. In spite of their improved tolerance, selective serotonin reuptake inhibitors do not appear to be particularly effective in the treatment of pain. Recently, a number of open and controlled trials with members of the new selective serotonin and noradrenaline reuptake inhibitor class of antidepressants, such as venlafaxine, milnacipran and duloxetine, suggest that these compounds may be more effective in relieving pain than selective inhibitors of serotonin reuptake. Wherever valid comparisons have been made the newer dual action drugs appear to be as effective as the tricyclics and considerably better tolerated. Dual action antidepressants are thus likely to become a widely used treatment of chronic pain both associated with and independent of depression.


G(i1) and G(oA) differentially determine kinetic efficacies of agonists for kappa-opioid receptor.
We examined the diversity of single receptor function by measuring receptor-G protein coupling in the baculovirus-Sf21 expression system. In comparative studies using Sf21 cell membranes expressing kappa-opioid receptor (KOR) plus Galpha(i1)beta(1)gamma(2) or KOR plus Galpha(oA)beta(1)gamma(2), there was no significant difference between both preparations in the K(i) values of various kappa-opioid ligands for the displacement of [(3)H]U69593 binding. However, a marked difference in the rank order of agonists to stimulate [(35)S]GTPgammaS binding was observed between both preparations. These findings suggest that agonist efficacy is dependent on the population of different G proteins expressed in various tissues.


Treatment of ulcerative colitis with an engineered human anti-TNFalpha antibody CDP571.
Tumour Necrosis Factor-alpha (TNFalpha) is a pro-inflammatory cytokine whose expression is increased in the colonic mucosa of patients with active ulcerative colitis. TNFalpha antibodies have been shown to be beneficial in animal models of bowel inflammation and in Crohn's disease but have not previously been studied in ulcerative colitis. Patients with mild/moderate ulcerative colitis were treated openly with a single intravenous infusion of 5 mg/kg of an engineered human IgGgamma4 antibody CDP571 and monitored for 8 weeks. Fifteen patients entered the study, eight males and seven females, with a mean age of 44 years. Eleven had left-sided disease, four extensive disease and six patients were steroid-unresponsive. The treatment was well tolerated and plasma half-life of CDP571 was approximately 7 days. There was a significant reduction from 6.7 to 4.6 (P = 0.023) in the mean Powell-Tuck score by 1 week post-infusion and a reduction to 5.5 was seen at 2 weeks (P = 0.218). Significant but modest reductions also occurred in erythrocyte sedimentation rate and serum C reactive protein in the first 2 weeks. Mean Interleukin-6 plasma concentrations fell from 6.9 to 5.4 pg/mL by week 1, and to 6.1 pg/mL by week 2 (NS). Reductions in sigmoidoscopic score and number of liquid stools were noted but failed to reach statistical significance. A consistent improvement in disease activity was seen in the initial 2 weeks after infusion and the treatment was well tolerated. These promising results support the testing of CDP571 in a larger controlled trial.


Induced down-regulation of neuropeptide Y-Y1 receptors delays initiation of kindling.
Neuropeptide Y appears to modulate epileptic seizures differentially according to the receptor subtypes involved. In the hippocampus, neuropeptide Y expression and release are enhanced in different models of epileptogenesis. On the contrary, the expression of Y1 receptors is decreased and it has been shown that activation of these receptors has pro-convulsant effects. The aim of our study was to investigate the role of Y1 receptors during hippocampal kindling epileptogenesis using (i) knock-out mice lacking Y1 receptors and (ii) intrahippocampal infusion of Y1 antisense oligodeoxynucleotide in rats. Y1 knock-out mice showed similar susceptibility to seizure induction and presented no difference in kindling development as compared with their control littermates. Conversely, local hippocampal down-regulation of Y1 receptors during the first week of hippocampal kindling, induced by a local infusion of a Y1 antisense oligodeoxynucleotide, significantly increased seizure threshold intensity and decreased afterdischarge duration. A reverse effect was observed during the week following the infusion period, which was confirmed by a significant decrease in the number of hippocampal stimulations necessary to evoke generalized seizures. At the end of this second week, an up-regulation of Y1 receptors was observed in kindled rats infused with the antisense as compared with the mismatch-treated controls. Our results in the rat suggest that the down-regulation of Y1 receptors in the hippocampus participates in the control of the initiation of epileptogenesis. The lack of an effect of the deficiency of Y1 receptors in the control of kindling development in Y1 knock-out mice could be due to compensatory mechanisms.


Effects of calcium antagonists on renal hemodynamics.
Recent attention has been focused on the effects of calcium antagonists on renal function. When administered in vitro to the isolated perfused kidney, calcium antagonists exhibit consistent actions permitting characterization of actions permitting characterization of their renal effects. Calcium antagonists do not affect the vasodilated isolated perfused kidney, but they do markedly alter the response of the kidney to vasoconstrictor agents. In the presence of norepinephrine, calcium antagonists markedly augment the glomerular filtration rate but produce only a modest improvement in renal perfusion. Studies using the isolated perfused hydronephrotic rat kidney model, which permits direct visualization of afferent and efferent arterioles, have demonstrated that the augmentation of the glomerular filtration rate is attributable to a preferential vasodilation of preglomerular vessels. Although the clinical implications of such observations are not fully delineated, preliminary studies in experimental animal models indicate that calcium antagonists might exert salutary effects on renal function in clinical settings characterized by an acute impairment of renal hemodynamics. It is apparent, however, that the renal hemodynamic effects of calcium antagonists commend their use in the management of essential hypertension.