[Epidemiological study on chronic renal failure elderly patients on hemodialysis].
Hemodialysis shows an increased prevalence in elderly patients, a population which often presents poor nutrition, high prevalence of cardiovascular, neurological and osteoarticular diseases and psycho-social problems. The objective of this epidemiological, cross-sectional and multicenter study, in patients older than 65 years (n 625) and >75 years (n 558) from 29 Spanish medical institutions was to perform an epidemiological analysis It included demographic information, as well as data regarding chronic renal failure, functional and psychological abilities (Katz Index, Lawton and Karnofsky Scales), dialysis logistics and clinical parameters. The study analyzed data from 1,183 patients (678 female), mean age 75.4+/-5.5 years; mean duration of dialysis 4.3+/-5.1 years (57.7% were referred by the GP: general practitioner). The most frequent etiologies were diabetic nephropathy (21.2%) and vascular renal disease (20.9%). The main comorbilites were high blood pressure (75.6%), Diabetes Mellitus (32.9%) and vascular (29.0%) and osteoarticular (27.3%) diseases. The great majority of patients lived at their family home (85.0%), travelled to their dialysis units alone (80.8%) and by ambulance (56.7%), and it took them less than an hour to arrive (87.5%). Over 75% of patients were fully functional (79.4% under 75 years and 71.6% over 75); meanwhile 10.5% were partially impaired and 13,8% severely impaired. Karnofsky performance scale scored less than 70 in 59.4% of the patients. Analytical parameters rated Hb >or= 11 g/dL for 81.7% of patients; ferritin >or= 100 ng/dL for 98.5%; PTH 150-300 pg/mL for 31.9%; albumin >3.5 g/dL for 75.6%; and serum phosphor <5.5 mg/dL for 70.6%. For the dialysis Kt/V the mean value was 1.4+/-0.3 with a mean duration of dialysis session of 11.7+/-4.0 hours/week. High permeability membranes were used in 52.3% of patients and internal arteriovenous fistula in 74.0%. Around 75% of elderly patients on hemodialysis fulfill age-suitable daily living activities and display adequate dialysis quality parameters.


RNA-Dependent RNA Polymerase Speed and Fidelity are not the Only Determinants of the Mechanism or Efficiency of Recombination.
Using the RNA-dependent RNA polymerase (RdRp) from poliovirus (PV) as our model system, we have shown that Lys-359 in motif-D functions as a general acid in the mechanism of nucleotidyl transfer. A K359H (KH) RdRp derivative is slow and faithful relative to wild-type enzyme. In the context of the KH virus, RdRp-coding sequence evolves, selecting for the following substitutions: I331F (IF, motif-C) and P356S (PS, motif-D). We have evaluated IF-KH, PS-KH, and IF-PS-KH viruses and enzymes. The speed and fidelity of each double mutant are equivalent. Each exhibits a unique recombination phenotype, with IF-KH being competent for copy-choice recombination and PS-KH being competent for forced-copy-choice recombination. Although the IF-PS-KH RdRp exhibits biochemical properties within twofold of wild type, the virus is impaired substantially for recombination in cells. We conclude that there are biochemical properties of the RdRp in addition to speed and fidelity that determine the mechanism and efficiency of recombination. The interwoven nature of speed, fidelity, the undefined property suggested here, and recombination makes it impossible to attribute a single property of the RdRp to fitness. However, the derivatives described here may permit elucidation of the importance of recombination on the fitness of the viral population in a background of constant polymerase speed and fidelity.


Depression during treatment with beta-blockers: results from a double-blind placebo-controlled study.
Depressive symptoms were rated during a double-blind placebo-controlled trial of nadolol for chronic aggression. Depressive symptoms were not significantly different in nadolol and placebo groups during any phase of the drug trial.


Laboratory diagnosis of amebiasis in a sample of students from southeastern Brazil and a comparison of microscopy with enzyme-linked immunosorbent assay for screening of infections with Entamoeba sp.
Epidemiological studies on amebiasis have been reassessed since Entamoeba histolytica and E. dispar were first recognized as distinct species. Because the morphological similarity of these species renders microscopic diagnosis unreliable, additional tools are required to discriminate between Entamoeba species. The objectives of our study were to compare microscopy with ELISA kit (IVD®) results, to diagnose E. histolytica infection, and to determine the prevalence of amebiasis in a sample of students from southeastern Brazil. In this study, diagnosis was based on microscopy due to its capacity for revealing potential cysts/trophozoites and on two commercial kits for antigen detection in stool samples. For 1,403 samples collected from students aged 6 to 14 years who were living in Divinópolis, Minas Gerais, Brazil, microscopy underestimated the number of individuals infected with E. histolytica/E. dispar (5.7% prevalence) compared with the ELISA kit (IVD®)-based diagnoses (15.7% for E. histolytica/E. dispar). A comparison of the ELISA (IVD®) and light microscopy results returned a 20% sensitivity, 97% specificity, low positive predictive value, and high negative predictive value for microscopy. An ELISA kit (TechLab®) that was specific for E. histolytica detected a 3.1% (43/1403) prevalence for E. histolytica infection. The ELISA kit (IVD®) can be used as an alternative screening tool. The high prevalence of E. histolytica infection detected in this study warrants the implementation of actions directed toward health promotion and preventive measures.


Colocalization of dopamine and serotonin in the rat pituitary gland and in the nuclei innervating it.
The nerve terminals in the intermediate and posterior lobes of the rat pituitary gland are reported to show colocalization of serotonin and tyrosine hydroxylase. This study examined the extent of this colocalization in the pituitary gland and in the nuclei considered to project to the pituitary. In the intermediate lobe, two types of nerve fibers were encountered, one containing serotonin (5-HT-IR) and tyrosine hydroxylase (TH-IR) immunoreactivities and the other showing 5-HT-IR only. Instead, there was no colocalization in the posterior lobe. In the hypothalamus, colchicine treatment with L-tryptophan and pargyline injections resulted in 5-HT-IR in some neurons in the dorsomedial, periventricular and arcuate nuclei, some of which in the arcuate and periventricular nuclei were also TH-IR. In the raphe nuclei no colocalization of 5-HT-IR and TH-IR was observed. Catecholamine neurotoxin, 6-hydroxydopamine, abolished the 5-HT-IR and dramatically reduced the TH-IR in the intermediate lobe nerve fibers. Both effects were prevented by cocaine, a monoamine uptake inhibitor, but not by fluoxetine, a specific serotonin uptake inhibitor. Serotonin neurotoxin p-chloroamphetamine (PCA) had no effect on intermediate lobe fibers, although it caused complete disappearance of 5-HT-IR from the posterior lobe nerve fibers. This effect was prevented by fluoxetine. Our results indicate, that colocalization of serotonin and TH observed in the intermediate lobe occurs both in the nerve terminals within the lobe and in some nuclei that innervate it. Furthermore, drug treatments suggest that serotonin in the intermediate lobe is localized in catecholaminergic fibers, which do not posses a specific serotonin uptake mechanism.


Oral dexamethasone attenuates Doxil-induced palmar-plantar erythrodysesthesias in patients with recurrent gynecologic malignancies.
The aim of this study is to evaluate the effect of oral dexamethasone in attenuating palmar-plantar erythrodysesthesias (PPE) in Doxil-treated patients for gynecologic malignancies. An IRB-approved prospective case study was conducted in patients with recurrent gynecologic malignancies who were treated with Doxil (50 mg/m(2)) on a 28-day cycle. Patients experiencing grades II-IV PPE were delayed until resolution then retreated without dose reduction and with a tapering oral dexamethasone regimen (8 mg BID days -1 to 4; 4 mg BID day 5; 4 mg day 6). Standard treatment for grades II-IV PPE in those not receiving dexamethasone was weekly dose delay until resolution of symptoms up to 2 weeks. If resolution occurred within 3 weeks of delay, a 25% dose reduction was made. Persistent grades III/IV PPE resulted in withdrawal of Doxil. Twenty-three patients (ovarian-16, uterine-7) were treated between January 1998 and December 2000. The median number of cycles administered was 5 (range 1-20). Nine patients (39%) developed grades II-IV PPE. All nine patients received more than five cycles of Doxil. The median time to PPE was 3 cycles (range 2-5). Six out of nine PPE patients received scheduled dose dexamethasone. All six had complete or near complete resolution of PPE and all continued treatment without subsequent dose modification. All three of the nine PPE patients not receiving dexamethasone required treatment delays and were dose reduced. Oral dexamethasone is effective in attenuating or eliminating Doxil-induced PPE. The use of the dexamethasone regimen prevents treatment delay and dose reduction.


[Pathogenicity factors of various vibrios and aeromonads].
A review of literature on the pathogenicity factors of non-cholera vibrios and Aeromonas is presented. A detailed analysis of such pathogenicity factors as hemolysins, lecithinases, cytotoxins, adhesins, etc. is given. Information on some mechanisms of the pathogenetic action of these factors on warm-blooded animals is presented. The necessity of more extensive research on pathogenicity and persistence factors in opportunistic bacteria causing sapronotic infections is emphasized.


Pharmacological inhibition of phosphatidylcholine biosynthesis is associated with induction of phosphatidylinositol accumulation and cytolysis of neoplastic cell lines.
De novo production of phosphatidic acid (PA) in tumor cells is required for phospholipid biosynthesis and growth of tumor cells. In addition, PA production by phospholipase D has been cited among the effects of certain oncogenes and growth factors. In this report, it has been demonstrated that enhanced phospholipid metabolism through PA in tumor cells can be exploited pharmacologically for development of anticancer agents, such as CT-2584, a cancer chemotherapeutic drug candidate currently in Phase II clinical trials. By inhibiting CTP:choline-phosphate cytidylyltransferase (CT), CT-2584 caused de novo phospholipid biosynthesis via PA to be shunted away from phosphatidylcholine (PC) and into phosphatidylinositol (PI), the latter of which was doubled in a variety of CT-2584-treated tumor cell lines. In contrast, cytotoxic concentrations of cisplatin did not induce accumulation of PI, indicating that PI elevation by CT-2584 was not a general consequence of chemotherapy-induced cell death. Consistent with this mechanism of action, propranolol, an inhibitor of PA phosphohydrolase and phosphatidylcholine biosynthesis, was also cytotoxic to tumor cell lines, induced PI accumulation, and potentiated the activity of CT-2584 in cytotoxicity assays. As expected from biophysical properties of anionic phospholipids on cellular membranes, CT-2584 cytotoxicity was associated with disruption and swelling of endoplasmic reticulum and mitochondria. We conclude that CT-2584 effects a novel mechanism of cytotoxicity to cancer cells, involving a specific modulation of phospholipid metabolism.


Expression of Dicer in rheumatoid arthritis is associated with disease activity and balances the production of TNF‑α.
Gene expression can be altered through RNA interference (RNAi), including microRNA (miRNA) or small interfering RNA. Alterations of miRNAs in rheumatoid arthritis (RA) have been reported, however, the components of the RNAi machinery in RA remain to be fully elucidated. The aim of the present study was to detect the levels of Dicer, Argonaute2 and Drosha, components of the RNAi machinery, in the peripheral blood mononuclear cells of patients with RAusingreverse transcription‑quantitative polymerase chain reaction, and to compare the results with disease activity and clinical features. Disease activity was assessed using the Disease Activity Score 28 (DAS28). Transfection and stimulation of cultured cells were conducted to determine the biological function of Dicer. ELISA was used to test tumor necrosis factor (TNF)‑α protein levels. It was found that the mRNA expression levels of Dicer and Drosha were upregulated in patients with RA, and that the increased level of Dicer was correlated with disease activity in patients with RA. Dicer and TNF‑α were activated in the serum of patients with RA. The activation of Dicer suppressed the production of TNF‑α. These results suggested that Dicer can balance the production of TNF‑α, and thus may serve as a regulator of the immune response in patients with RA.


Neocortical synaptic proliferation following forebrain-dependent trace associative learning.
Many behavioral studies have suggested that learning induces neocortical synaptic modifications. However, neocortical synaptic modifications following forebrain-dependent trace associative learning has not been closely examined. Acquisition of whisker-trace-eyeblink (WTEB) conditioning, a forebrain-dependent trace associative task, has been reported to modulate the expression of cytochrome oxidase, a marker for metabolic activity, in the conditioned barrels, suggesting that trace associative conditioning induces neocortical synaptic plasticity. However, neocortical synaptic plasticity has never been directly examined following this trace associative task. To assess neocortical synaptic modifications, the present study examined synapsin I expression following WTEB conditioning. Synapsin I is part of a phosphoprotein family involved in neuronal regulation of neurotransmitter release that also exhibits an expression pattern closely correlating to synapse number. Findings from this study demonstrated that synapsin I expression is elevated in primary somatosensory neocortex in trace-paired-conditioned mice compared with unpaired-conditioned (stimulation-control) mice and naïve mice, suggesting that WTEB conditioning induces synaptic proliferation. Additional findings from the present study examining cytochrome oxidase expression replicated previous findings demonstrating that WTEB conditioning induces a learning-specific expansion of the cytochrome oxidase staining expression for conditioned barrels. Together, these results suggest that synaptic proliferation is contributing to the learning-induced metabolic augmentation previously observed in conditioned barrels following WTEB conditioning. Furthermore, these results suggest that trace associative learning facilitates neocortical synaptic modification.


Cellular and humoral immunity to the 60-kD heat shock protein in inflammatory bowel disease.
Mycobacteria have been considered a possible etiological agent in Crohn's disease. Since cross-reactivity between epitopes of mycobacterial and self-heat shock protein might represent a potential disease mechanism, we determined the cellular and humoral immune responses to the mycobacterial and the human 60-kD heat shock protein, as well as various control antigens. We studied samples from 19 patients with Crohn's disease, 12 patients with ulcerative colitis, and from 19 healthy individuals. T cell responses were studied using a standard proliferation assays to purified recombinant mycobacterial and human 60-kD heat shock protein. Antibody levels were measured by establishing an enzyme-linked immunosorbent assays to recombinant purified 60-kD heat shock protein. The proliferative responses to the mycobacterial and the human 60-kD heat shock protein were closely correlated, but the cellular immune response to both antigens was not significantly different between the two patient groups and healthy controls. In addition, antibody levels to heat shock protein were also not significantly different between the three groups. These results argue strongly against a role for mycobacterial heat shock protein in the pathogenesis of Crohn's disease. However, in view of previous data demonstrating overexpression of 60-kD heat shock protein in inflammatory Crohn's disease tissue, this considerable cellular and humoral autoreactivity suggests an important immunoregulatory role for the 60-kD heat shock protein.


EED and EZH2 constitutive variants: A study to expand the Cohen-Gibson syndrome phenotype and contrast it with Weaver syndrome.
Overgrowth-intellectual disability (OGID) syndromes are characterized by increased growth (height and/or head circumference ≥+2 SD) in association with an intellectual disability. Constitutive EED variants have previously been reported in five individuals with an OGID syndrome, eponymously designated Cohen-Gibson syndrome and resembling Weaver syndrome. Here, we report three additional individuals with constitutive EED variants, identified through exome sequencing of an OGID patient series. We compare the EED phenotype with that of Weaver syndrome (56 individuals), caused by constitutive EZH2 variants. We conclude that while there is considerable overlap between the EED and EZH2 phenotypes with both characteristically associated with increased growth and an intellectual disability, individuals with EED variants more frequently have cardiac problems and cervical spine abnormalities, boys have cryptorchidism and the facial gestalts can usually be distinguished.


Interspecies differences in the accumulation of tributyltin and its metabolites under dietary exposure in sea perch, Lateolabrax japonicus, and red sea bream, Pagrus major.
Interspecies differences in the accumulation of dietary tributyltin (TBT) between sea perch, Lateolabrax japonicus, and red sea bream, Pagrus major, were studied. Although TBTs in both species reached steady-state condition in low-concentration group (L-group, 200 ng/g nominal concentration) by 1 week, it increased up to the end of exposure in high-concentration group (H-group, 3000 ng/g nominal concentration). In H-group, the accumulation rate of TBT in sea perch from 1 to 2 weeks was much higher than in red sea bream, which were 2.4-fold for sea perch and 1.7-fold for red sea bream, although TBT concentrations were similar between sea perch and red sea bream in L-group. Furthermore, in the H-group, the concentrations of TBT at 1 and 2 weeks were about 1.3- and 1.9-fold, respectively, higher in the sea perch than in the red sea bream. On the other hand, DBT residue in red sea bream was about 1.4-fold higher in sea perch for the L-group but concentrations were similar in both fishes for the H-group. These results suggest that red sea bream could metabolize faster the TBT to DBT than sea perch. This study also reveals that fish probably could absorb TBT through the food chain. The uptake of TBT by fish should be regarded in the real environment, because TBT concentration in seawater has been decreasing and now already at significantly low level.


Novel antifungal agents as salvage therapy for invasive aspergillosis in patients with hematologic malignancies: posaconazole compared with high-dose lipid formulations of amphotericin B alone or in combination with caspofungin.
In patients with hematologic malignancy, invasive aspergillosis continues to be associated with high mortality even when treated with conventional antifungal therapy. To investigate novel antifungal agents, we compared 53 patients who received posaconazole salvage therapy to 52 contemporary control patients who received high-dose lipid formulation of amphotericin B (HD-LPD/AMB at > or = 7.5 mg kg(-1) per day) and 38 other control patients who received caspofungin plus HD-LPD/AMB. Patients in the three groups had similar. The overall response rate to salvage therapy was 40% for posaconazole, 8% for HD-LPD/AMB (P < or = 0.001) and 11% for combination therapy (P < 0.002). Aspergillosis contributed to the death of 40% of posaconazole group, 65% of the HD-LPD/AMB group and 68% of the combination group (P < or = 0.008). By multivariate analysis, posaconazole therapy independently improved response (9.5; 95% confidence interval, 2.8-32.5; P < 0.001). HD-LPD/AMB alone or in combination was associated with a significantly higher rate of nephrotoxicity (P < or = 0.02) and hepatotoxicity (P < 0.03). In conclusion, posaconazole salvage therapy demonstrated greater efficacy and safety than HD-LPD/AMB alone or in combination with caspofungin in the salvage therapy of invasive aspergillosis in hematologic malignancy.


Characterization of the phytopathogen Pseudomonas syringae pathovar ribicola NCPPB 963.
In 1939, a bacterial spot caused severe defoliation of Ribes aureum (Golden Currant) The causal agent is now recognized as Pseudomonas syringae pathovar ribicola. This communication extends the phenotype of the only identified strain of P. syringae pv. ribicola, which is reminiscent of those of other pathovars, and provides a molecular biological characterization. A minimum size of 5.55 Mb for the bacterial genome was obtained using pulsed-field electrophoresis. The SDS-PAGE outer-membrane profile contained seven major bands, and has obvious similarities to that of P. aeruginosa. SDS-PAGE of concentrated mid-log phase culture supernatants revealed large amounts of a single, cryptic 24.0 kD protein. The amino acid composition and 57 residues in the N-terminus of this protein. were determined. The protein sequence was nearly identical to the translation of a region of unknown function in the P. aeruginosa genome. Extensive similarity in N-terminal sequence, composition and subunit size to a secreted hydrophilic Vibrio cholerae protein of unknown function was also found. Neither protein has been directly associated with disease development.


A urokinase-type plasminogen activator-inhibiting cyclic peptide with an unusual P2 residue and an extended protease binding surface demonstrates new modalities for enzyme inhibition.
To find new principles for inhibiting serine proteases, we screened phage-displayed random peptide repertoires with urokinase-type plasminogen activator (uPA) as the target. The most frequent of the isolated phage clones contained the disulfide bridge-constrained sequence CSWRGLENHRMC, which we designated upain-1. When expressed recombinantly with a protein fusion partner, upain-1 inhibited the enzymatic activity of uPA competitively with a temperature and pH-dependent K(i), which at 25 degrees C and pH 7.4 was approximately 500 nm. At the same conditions, the equilibrium dissociation constant K(D), monitored by displacement of p-aminobenzamidine from the specificity pocket of uPA, was approximately 400 nm. By an inhibitory screen against other serine proteases, including trypsin, upain-1 was found to be highly selective for uPA. The cyclical structure of upain-1 was indispensable for uPA binding. Alanine-scanning mutagenesis identified Arg(4) of upain-1 as the P(1) residue and indicated an extended binding interaction including the specificity pocket and the 37-, 60-, and 97-loops of uPA and the P(1), P(2), P(3)', P(4)', and the P(5)' residues of upain-1. Substitution with alanine of the P(2) residue, Trp(3), converted upain-1 into a distinct, although poor, uPA substrate. Upain-1 represents a new type of uPA inhibitor that achieves selectivity by targeting uPA-specific surface loops. Most likely, the inhibitory activity depends on its cyclical structure and the unusual P(2) residue preventing the scissile bond from assuming a tetrahedral geometry and thus from undergoing hydrolysis. Peptide-derived inhibitors such as upain-1 may provide novel mechanistic information about enzyme-inhibitor interactions and alternative methodologies for designing effective protease inhibitors.


Dead-time correction in dynamic radionuclide studies by computer.
Several methods for the measurement of dead time and for evaluation of the dependencies required to correct dynamic studies for dead-time losses are described. Two program variants were written to produce time-activity curves in the selected areas of interest of the dynamic studies with computer's correction of dead-time losses; these variants are part of the system of programs for complex processing of scintigraphic studies set up for a Clincom instrument. The first variant performs correction on the basis of the registered count rate in the whole image, while the second variant makes us of the additional reference source positioned on the periphery of the camera visual field.


Effect of additives on the digestibility of corn stover solids following pretreatment by leading technologies.
Bovine serum albumin (BSA), Tween-20, and polyethylene glycol (PEG6000) were added to washed corn stover solids produced by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), dilute sulfuric acid (DA), lime, controlled pH, and sulfur dioxide (SO(2)) pretreatments and to untreated corn stover (UT) and pure Avicel glucan prior to adding cellulase supplemented with beta-glucosidase at an activity ratio of 1:2/g and a moderate enzyme loading of 16.1 mg/g glucan in the raw corn stover. The additives were applied individually at 150, 300, and 600 mg/g glucan in the pretreated solids and in combinations of equal amounts of each that totaled 600 mg/g. The greatest increase in total sugar release was by Tween-20 with SO(2) pretreated solids followed by PEG6000 with ARP solids and Tween-20 with lime solids. The effectiveness of the additives was observed to depend on the type of sugars left in the solids, suggesting that it may be more beneficial to use the mixture of these additives to realize a high total sugar yield. In addition, little enhancement in sugar release was possible beyond a loading of 150 mg additives/g glucan for most pretreatments, and combinations did not improve sugar release much over use of additives alone for all except SO(2). Additives were also found to significantly increase concentrations of cellobiose and cellooligomers after 72 h of Avicel hydrolysis.


Approaches to academic growth assessment.
There is much interest in assessing growth in student learning. Assessments of growth have important implications and affect many policy decisions at many levels. In the present article, we review some of the different approaches to measuring growth and examine the implications of their usage. Samples used in research on growth models typically include students enrolled in public schools that primarily serve kindergarten through the 12th grade. Definitions of growth and gain are reviewed, and five types of growth models are examined: (1) Student Gain Score Model, (2) The Covariate Adjustment Model, (3) The Student Percentile Gain Model - referred to as single-wave value-added models, (4) Univariate Value-Added Response Models, and (5) Multivariate Value-Added Response Models. Modelling approaches are vastly different, whereas Student Gain Models are mathematically and conceptually simple, Multivariate Models are highly complex. Educators assessing growth must make critical decisions about measurement. The type of instrument that is selected and the type of analytic techniques selected are of great importance. Growth must be considered from technical, pedagogical, and policy perspectives.


Natural variation and selection in GmSWEET39 affect soybean seed oil content.
Soybean (Glycine max) is a major contributor to the world oilseed production. Its seed oil content has been increased through soybean domestication and improvement. However, the genes underlying the selection are largely unknown. The present contribution analyzed the expression patterns of genes in the seed oil quantitative trait loci with strong selective sweep signals, then used association, functional study and population genetics to reveal a sucrose efflux transporter gene, GmSWEET39, controlling soybean seed oil content and under selection. GmSWEET39 is highly expressed in soybean seeds and encodes a plasma membrane-localized protein. Its expression level is positively correlated with soybean seed oil content. The variation in its promoter and coding sequence leads to different natural alleles of this gene. The GmSWEET39 allelic effects on total oil content were confirmed in the seeds of soybean recombinant inbred lines, transgenic Arabidopsis, and transgenic soybean hairy roots. The frequencies of its superior alleles increased from wild soybean to cultivated soybean, and are much higher in released soybean cultivars. The findings herein suggest that the sequence variation in GmSWEET39 affects its relative expression and oil content in soybean seeds, and GmSWEET39 has been selected to increase seed oil content during soybean domestication and improvement.


The in-vitro effect of antineoplastic agents on proliferative activity and cytoskeletal components of plaque-derived smooth-muscle cells from human coronary arteries.
Restenosis after successful percutaneous transluminal coronary angioplasty remains the major clinical problem limiting the long-term efficacy of the treatment. Recent advances in the understanding of the biology of restenosis indicate that its cause is predominantly a multifactorial stimulation of smooth-muscle cell proliferation. The aim of this study was to investigate the in-vitro effect of antineoplastic agents on smooth-muscle cells isolated from human coronary plaque material. Atherosclerotic tissue from coronary arteries was extracted from 15 patients of both sexes by thrombendarterectomy. Cells were isolated using enzymatic disaggregation and identified to be smooth-muscle cells with fluorescent antibodies for smooth-muscle-specific alpha-actin. The antineoplastic agents cytarabine (500-0.005 micrograms/ml), doxorubicin (50-0.0005 micrograms/ml), and vincristine (10-0.0001 micrograms/ml) were added to the cultures. Six days after seeding, the cells were trypsinized and then counted. All three antineoplastic agents had a strong dose-dependent antiproliferative effect on cultured smooth-muscle cells. After the application of cytostatic agents, cells either became rounded or underwent complete lysis. Cytoskeletal elements, such as actin, microtubules, and vimentin, were largely altered. This investigation examined the potential role of antineoplastic therapy in the prevention of restenosis after coronary angioplasty. The development of new intravascular delivery systems, such as coated stents, may open the way for local antiproliferative strategies in interventional cardiology.


Disaturated phosphatidylcholine in rat lungs with altered numbers of type II alveolar epithelial cells.
Rats exposed to 85 per cent oxygen for 9 days demonstrated an increase in lung deoxyribonucleic acid (DNA) specific acitvity for [3H]-thymidine and a significant increase in the volume of interstitial and alveolar epithelial cells. This included a 63 per cent increase in the number of Type II alveolar epithelial cells. Lamellar body volume per lung significantly increased in animals exposed to oxygen and was related to the increased number of Type II cells, because lamellar body volume expressed per Type II cell remained relatively constant and was comparable to control values. The concentration of whole lung disaturated phosphatidylcholine also increased significantly, but the concentration of unsaturated phosphatidylcholine did not increase. Concomitantly, there was a significant decrease in [32P]-orthophosphate and [14C]-palmitate disaturated phosphatidylcholine specific activity that was proportional to the increased number of alveolar epithelial Type II cells present.


Isolation of Cryptosporidium parvum and Cyclospora cayetanensis from vegetables collected in markets of an endemic region in Peru.
Cryptosporidium parvum and Cyclospora cayetanensis are protozoan pathogens that cause prolonged diarrhea in both immunocompetent and immunocompromised hosts. Cryptosporidium parvum can be transmitted via the fecal-oral route, while the exact mechanisms of transmission of Cyclospora cayetanensis have not been fully determined. Humans appear to be the sole host for the latter and a distinct seasonality has been observed in endemic areas around the world. Samples of vegetables were collected at several small markets in a periurban slum in Peru during the seasons of high and low incidence. The vegetables were washed, the supernatants were collected and centrifuged, and the pellets were resuspended in a solution of 2.5% potassium dichromate. Pellets were examined using direct microscopic observation, acid-fast staining, and immunofluorescent assays for C. parvum and Cyclospora cayetanensis oocysts. Samples were collected during three time periods: the season of low incidence, the beginning of the season of high incidence, and end of the season of high incidence. Of the total vegetables examined, 14.5% contained C. parvum oocysts and 1.8% had Cyclospora oocysts. Thus, market vegetables may provide a route by which Cryptosporidium and Cyclospora can be transmitted. Our study also suggests that washing vegetables does not completely remove Cryptosporidium and Cyclospora oocysts.


Harmful consequences of specific substances on the oral health.
Numerous substances may have an adverse influence on the oral region by virtue of their potentially harmful effects. Dental practitioners are often the first to see these effects in the head and neck region. Knowledge of the damaging consequences of agents such as alcohol, tobacco, areca nut, medications, alcohol-containing mouthwashes, dental devices and materials can assist the practitioners in clinical decision-making and accurate diagnosis of associated lesions. More importantly, timely diagnosis and prompt reporting of such harmful effects improve the quality of care. Such knowledge may also help in patient's education for the avoidance of associated adverse effects (AEs). Consequently, a thorough acquaintance of AEs will consolidate the distinct role of medical and oral health practitioners in safeguarding the patients' oral and systemic well-being. This article was planned to review the harmful consequences of specific substances on the oral health. Traditionally, the emphasis of review articles has been on reviewing the harmful effects of a single entity on oral health. This review is unique in the sense that it comprehensively and collectively focussed on the harmful effects on the oral health of addictive and illicit substances (alcohol, betel nut), medications, alcohol-containing mouthwashes, medications and dental materials.


Characteristics of particle coagulation in an underground parking lot.
Particles in vehicle exhaust plumes in underground parking lots have adverse health effects due to the enclosed environment in which they are released and the temperature difference between the tailpipe and ambient environment; at the same time, particle coagulation might be obvious near the tailpipe in an underground parking lot. In the present study, airflow and temperature fields were calculated using the Realizable k-ε model, and the Eulerian particle transport model was selected in the numerical simulation of particle concentration dispersion. Polydisperse thermal coagulation due to Brownian collisions was employed to calculate the particle coagulation. The results show that particle coagulation rate and half-time were significant within 1 m from the tailpipe. The variations in the particle coagulation rate and half-time were similar, but their directions were opposite. Air exhaust time was nearly four times longer than averaged half-time and 40 times longer than minimum half-time. The peak particle diameter increased approximately 1.43 times due to coagulation. A double particle concentration at the tailpipe caused the fourfold rise in the particle coagulation rate in the distance ranging less than 1 m from the tailpipe. An increase in exhaust velocity at the tailpipe could shorten the obvious range of particle coagulation along the centerline of the tailpipe from 1 to 0.8 m in the study.


Evaluation of automated time-lapse microscopy for assessment of in vitro activity of antibiotics.
This study aimed to evaluate the potential of a new time-lapse microscopy based method (oCelloScope) to efficiently assess the in vitro antibacterial effects of antibiotics. Two E. coli and one P. aeruginosa strain were exposed to ciprofloxacin, colistin, ertapenem and meropenem in 24-h experiments. Background corrected absorption (BCA) derived from the oCelloScope was used to detect bacterial growth. The data obtained with the oCelloScope were compared with those of the automated Bioscreen C method and standard time-kill experiments and a good agreement in results was observed during 6-24h of experiments. Viable counts obtained at 1, 4, 6 and 24h during oCelloScope and Bioscreen C experiments were well correlated with the corresponding BCA and optical density (OD) data. Initial antibacterial effects during the first 6h of experiments were difficult to detect with the automated methods due to their high detection limits (approximately 105CFU/mL for oCelloScope and 107CFU/mL for Bioscreen C), the inability to distinguish between live and dead bacteria and early morphological changes of bacteria during exposure to ciprofloxacin, ertapenem and meropenem. Regrowth was more frequently detected in time-kill experiments, possibly related to the larger working volume with an increased risk of pre-existing or emerging resistance. In comparison with Bioscreen C, the oCelloScope provided additional information on bacterial growth dynamics in the range of 105 to 107CFU/mL and morphological features. In conclusion, the oCelloScope would be suitable for detection of in vitro effects of antibiotics, especially when a large number of regimens need to be tested.


Determination of minocycline in human plasma by high-performance liquid chromatography coupled to tandem mass spectrometry: application to bioequivalence study.
Minocycline was determined in human plasma by HPLC-MS-MS using clarithromycin as an internal standard. The method is fast (single liquid extraction and run time of <3 min) and sensitive (5 ng/ml) and it was employed in a bioequivalence study of two 100 mg tablet formulations in 24 healthy volunteers. The 90% confidence interval of the individual ratio geometric mean for both AUC(0-96 h) and Cmax were 99.2-111.1% and 95.6-117.5%, respectively. Thus, Minoderm was considered bioequivalent to Minomax according to both the rate and extent of absorption. No food interaction was observed with either formulation.


Typical pollutants in bottom ashes from a typical medical waste incinerator.
Incineration of medical waste (MW) is an important alternative way for disposal of this type of hazardous waste, especially in China because of the outbreak of severe acute respiratory syndromes (SARs) in 2003. Thus, far, fly ash has received much attention but less attention has been paid to bottom ash. In this study, bottom ash samples were collected from a typical MW incinerator, and typical pollutants including heavy metals and polycyclic aromatic hydrocarbons (PAHs) in the ash were examined. X-ray fluorescence spectroscopy results indicated that CaO, SiO(2) and Al(2)O(3) were the main components of the bottom ash. Inductively coupled plasma-optical emission spectroscopy showed that the ash contained large amounts of heavy metals, including Zn, Ti, Ba, Cu, Pb, Mn, Cr, Ni and Sn. Most of the heavy metals (e.g., Ba, Cr, Ni, and Sn) presented in the residual fraction; whereas Mn, Pb and Zn presented in Fe-Mn oxides fraction, and Cu in organic-matter fraction. Toxicity characteristic leaching procedure tests indicated that the leached amounts of heavy metals were well below the limits. The sum of 16 US EPA priority PAHs (Sigma PAHs) varied from 10.30 to 38.14 mg kg(-1), and the total amounts of carcinogenic PAHs ranged between 4.09 and 16.95 mg kg(-1), exceeding the limits regulated by several countries. This research provides basic information for the evaluation of the environmental risk of MW incinerator bottom ash.


Molecular mechanistic pathway of colorectal carcinogenesis associated with intestinal microbiota.
The colon rectal portion of gastrointestinal tract (GI) is full of microorganisms with different complex community that plays important role in maintaining homeostasis. But now-a-days different literature indicated that microbiota cause development of colorectal cancer (CRC) with a disease and ultimately aggravates to death. The mechanism inside the colo-rectal portion of GI tract is not fully well-known and bacterial contribution inside it is also fully unclear. Therefore, there is certain evidence trying a target about the unclear mechanism between intestinal microbiota and CRC. Different reports revealed that colo-rectal microorganisms is playing a great role in inducing the onset and progression of CRC with different dynamic mechanisms viz. acceleration of chronic inflammatory state, the biosynthesis of genotoxins that interfere with cell cycle regulation, the production of toxic metabolites, or heterocyclic amine activation of pro-diet carcinogenic compounds. There is growing evidence that individuals with colonic adenomas and carcinomas harbor a distinct microbiota. Alterations to the gut microbiota may allow the outgrowth of bacterial populations that induce genomic mutations or exacerbate tumor-promoting inflammation. While cancer is largely considered to be a disease of genetic and environmental factors, increasing evidence has demonstrated a role for the microbiota in shaping inflammatory environments and promoting tumor growth and spread. Despite all these advances, different studies depicted the relationship between microbiota and CRC in humans and animal models and aid in developing alternate therapeutic approach based on gut microbiota manipulations. Alteration of the microbiota may be a useful to preventing and altering the trajectory of colorectal cancer. Therefore, the aim of the study is to identify the possible mechanistic mechanism regarding host-microbiota interaction in colorectal carcinogenesis.


[Perspectives in the inhalative administration of drugs].
Administration of drugs via the inhalation route will find new indications in the therapy for lung diseases. Furthermore, aerosolised drugs are of increasing interest for systemic treatment. The inhalation of antibiotics is already a well established therapy in cystic fibrosis. In bronchiectasis, severe COPD with bacterial airway colonisation and in mechanically ventilated patients, aerosolised aminoglycosides may provide benefit. Substitution of alpha1-antitrypsin in lung emphysema via inhalation seems to be superior to the intravenous administration. Inhaled insulin has currently been withdrawn from the market in spite of being approved by FDA and EMEA. However, intensive scientific research is still ongoing and many clinical studies are underway in the field of the inhalation of insulin, heparins and other systemic treatments.


An analysis of atmospheric water vapor variations over a complex agricultural region using airborne imaging spectrometry.
Understanding atmospheric water vapor patterns can inform regional understanding of water use, climate patterns and hydrologic processes. This research uses Airborne Visible Infrared Imaging Spectrometer (AVIRIS) reflectance and water vapor imagery to investigate spatial patterns of water vapor in California's Central Valley on a June date in 2013, and 2015, and relates these patterns to surface characteristics and atmospheric properties. We analyze water vapor imagery at two scales: regional and agricultural field, to examine how the slope, intercept, and trajectory of water vapor interact with the landscape in a highly diverse and complex agricultural setting. At the field scale, we found significant quadratic relationships between water vapor slope and wind magnitude in both years (p<0.001). Results showed a positive correlation between crop water use and the frequency with which crops showed directional agreement between wind and water vapor (r = 0.23). At the regional scale, we found patterns of water vapor that indicate advection of moisture across the scene. Water vapor slope was inversely correlated to field size with correlations of -0.37, and -0.28 for 2013 and 2015. No correlation was found between green vegetation fraction and vapor slope (r = 0.001 in 2013, r = 0.02 in 2015), but a weak correlation was found for the intercept (r = 0.11 in 2013, r = 0.26 in 2015). These results lead us to conclude that accumulation of water vapor above fields in these scenes is observable with AVIRIS-derived water vapor imagery whereas advection at the field level was inconsistent. Based on these results, we identify new opportunities to use and apply water vapor imagery to advance our understanding of hydro-climatic patterns and applied agricultural water use.


Women treated for epilepsy during pregnancy: outcomes from a nationwide population-based cohort study.
Women with epilepsy (WWE) are generally treated as a risk group during pregnancy, but over 90% of pregnant WWE have favorable pregnancies. However, the risk of some pregnancy and delivery complications may be increased among WWE, especially those on antiepileptic drugs. This nationwide, retrospective population-based cohort study includes WWE who gave birth in Finland during 1987-2008 (n = 1737) and the reference cohort of a random sample of women without epilepsy (n = 4357). Identification of the cohorts, and information on hospitalizations and deliveries were obtained from the Finnish Health Registers and population statistics. Multivariate analyses were conducted by binomial regression. WWE were more often hospitalized during pregnancy for accidents or other external causes [adjusted risk ratio (aRR) 1.74, 95% confidence interval (CI) 0.98-3.09], premature rupture of membranes (aRR 1.75, 95% CI 1.14-2.69) and premature contractions (aRR 1.75, 95% CI 1.36-2.23). Hospitalizations for infections were more frequent in WWE (1.4% vs. 0.4%, aRR 3.15, 95% CI 1.72-5.76). The risk for induction of delivery or a cesarean section was increased in WWE. There was no difference in premature deliveries between the groups, but the risk of being small for gestational age (aRR 1.57, 95% CI 1.23-2.01), admission to neonatal intensive care unit (aRR 1.66, 95% CI 1.39-1.97), and need for respiratory care (aRR 2.37, 95% CI 1.57-3.60) was clearly increased in the offspring of WWE. WWE are at an increased risk of complications and hospitalizations during pregnancy and delivery. However, the majority of WWE have a normal pregnancy and delivery.


Transcytosis of F4 fimbriae by villous and dome epithelia in F4-receptor positive pigs supports importance of receptor-dependent endocytosis in oral immunization strategies.
Very few antigens have been described that induce an intestinal immunity when given orally. Our laboratory demonstrated that oral administration of isolated F4 (K88) fimbriae of Escherichia coli to F4-receptor positive (F4R(+)) pigs induces protective mucosal immunity against challenge infection. However, presence of F4-receptors (F4R) on villous enterocytes is a prerequisite for inducing the immune response, as no F4-specific antibody-secreting cells (ASC) can be induced in F4R(-) pigs. In this study, the in vivo binding of isolated F4 fimbriae (F4) to the gut epithelium was examined in F4R(+) and F4R(-) pigs. It was further investigated whether binding of F4 to the F4R results in endocytosis in and translocation across the gut epithelium using microscopy. F4 did not adhere to the intestinal epithelium of F4R(-) pigs, whereas it strongly adhered to the villous epithelium and the follicle-associated epithelium (FAE) of the jejunum and ileum of F4R(+) pigs. Following binding to F4R, F4 was endocytosed by villous enterocytes, follicle-associated enterocytes and M cells. Transcytosis of F4 across the epithelium resulted in the appearance of F4 in the lamina propria and dome region of the jejunal and ileal PP. This is the first study showing transcytosis of fimbriae across the gut epithelium. This receptor-dependent transcytosis can explain the success of F4 fimbriae as oral immunogen for inducing protective immunity in F4R(+) pigs strengthening the importance of receptor-dependent endocytosis and translocation in oral vaccine strategies. Further identification of the receptor responsible for this transport is in progress.


Teaching and Learning: from staff nurse to nurse consultant. Part 7: teaching a skill.
I am always fascinated to watch people who are very competent at a particular task performing that skill. There is a confident ease that marks out a skilled performer but if you look deeper below the actual physical movements you can observe that the almost intuitive actions are guided by visual cues and sensory feedback. However, it is one thing to perform a skill but it is another to teach it.


Doping dependence of the nonmagnetic impurity resonance in cuprate superconductors.
Motivated by the observation that the nonmagnetic impurity induced resonance strength is reduced with underdoping in Bi(2)Sr(2)Ca(Cu(1-y)Zn(y))(2)O(8+x), we study the doping dependence of the nonmagnetic impurity induced resonance in cuprates from the viewpoint of resonating valence bond (RVB) spin liquid theory. It is revealed that the pseudogap state in the Yang, Rice and Zhang ansatz (Yang et al 2006 Phys. Rev. B 73 174501; Rice et al 2012 Rep. Prog. Phys. 75 016502) does not support an impurity resonance state due to the overdamping of the quasiparticle. In the coexistence of the d-wave superconductivity and the RVB gap, our theory provides an alternative but consistent explanation for the reduction of the resonance strength with underdoping, as well as the absence of the resonance in the large gap region.


Induced perceptual grouping.
The term perceptual grouping is associated with classical principles such as similarity and proximity. This article reports induced perceptual grouping, a phenomenon that occurs when placement of a uniform set of items near a structured set induces grouping within the otherwise uniform set. For example, when items grouped pair-wise by similarity are placed near another set of unstructured items, an analogous pair-wise grouping links elements of the second set. Induced grouping affected reaction times in two different visual search tasks, with reaction times depending on whether the target properties were contained within a group or crossed group boundaries as defined by induced grouping due to similarity, proximity, or common fate. Induced grouping was reduced when grouping between the structured and unstructured sets was weakened by means of a common-region cue or decreased similarity. Induced grouping appears to reflect the computation of hierarchical structure in visual images.


Ena/VASP proteins regulate cortical neuronal positioning.
Development of the multilayered cerebral cortex involves extensive regulated migration of neurons arising from the deeper germinative layers of the mammalian brain. The anatomy and formation of the cortical layers has been well characterized; however, the underlying molecular mechanisms that control the migration and the final positioning of neurons within the cortex remain poorly understood. Here, we report evidence for a key role of Ena/VASP proteins, a protein family implicated in the spatial control of actin assembly and previously shown to negatively regulate fibroblast cell speeds, in cortical development. Ena/VASP proteins are highly expressed in the developing cortical plate in cells bordering Reelin-expressing Cajal-Retzius cells and in the intermediate zone through which newly born cells migrate. Inhibition of Ena/VASP function through retroviral injections in utero led to aberrant placement of early-born pyramidal neurons in the superficial layers of both the embryonic and the postnatal cortex in a cell-autonomous fashion. The abnormally placed pyramidal neurons exhibited grossly normal morphology and polarity. Our results are consistent with a model in which Ena/VASP proteins function in vivo to control the position of neurons in the mouse neocortex.


In vivo evaporation rate of benzyl alcohol from human skin.
The evaporation rate of benzyl alcohol from the human volar forearm under controlled conditions in vivo shows a similar dependence on airflow and time to that seen in earlier in vitro studies. After an initial time lag associated with the apparatus, evaporation rate over a 2-h time period post-dose was satisfactorily described by a single exponential decay with a rate constant proportional to airflow over the skin, nu. The cumulative percentage of dose evaporated after 2 h ranged from 16% at nu = 20 mL min(-1) to 52% at nu = 100 mL min(-1). The absorption rate constant determined by an analysis of the in vivo data was equivalent to that determined in vitro, whereas the evaporation rate constants were related by the inverse ratio of the headspace volumes. The latter finding suggests that a simple laminar flow model can satisfactorily describe evaporation in both systems over the range of airflows used.


Changes in guidelines for cardiopulmonary resuscitation and emergency cardiac care.
At the Fifth National Conference on Cardiopulmonary Resuscitation and Emergency Cardiac Care held in Dallas in February 1992, changes in the guidelines for basic life support and advanced cardiac life support were recommended. These changes are defined and explained in this article.


Serum factors influencing spontaneous rosette formation by lymphocytes of pregnant women.
The effect of sera from pregnant women on the percentage of spontaneous rosette-forming peripheral lymphocytes was investigated. Pregnancy lymphocytes displayed a significantly lower capacity to bind SRBC than control male lymphocytes. However, after an exhaustive washing, it was possible to demonstrate a significant increase of spontaneous rosettes formed by pregnancy lymphocytes. It was found that the incubation of pregnancy-washed lymphocytes with pregnancy but not homologous male serum restored to depressed levels the values of rosette-forming peripheral lymphocytes. This blocking activity was significantly higher with autologous serum than homologous pregnancy serum. Control lymphocytes were unaffected both by washing and by incubation with pregnancy sera. The blocking activity was found in the same ion-exchange chromatography fraction of pregnancy serum where paternal HLA antigens could be demonstrated, and was reproduced by a soluble HLA preparation from the husband's lymphocytes.


Dual Factors May Be Necessary for Development of Atopic March in Early Infancy.
The incidence of atopic diseases, including atopic dermatitis (AD), food allergies, allergic rhinitis, and asthma, has increased in recent decades, and currently affects approximately 20% of the population. Atopic march is the development of AD in infancy and subsequent food allergies, allergic rhinitis, and asthma in later childhood. Patients with infantile eczema may develop typical symptoms of AD, allergic rhinitis, and asthma at certain ages. Some patients' symptoms persist for several years, whereas others may have resolution with aging. Development of these diseases is strongly influenced by the following two factors: skin dysfunction caused by filaggrin mutations and development of colonization of microflora in early infancy. Filaggrin mutations predisposing to asthma, allergic rhinitis, and allergic sensitization, only in the presence of AD, strongly support the role of filaggrin in the pathogenesis of AD and in subsequent progression of the atopic march. Several studies have shown that development of colonization of microflora in early infancy might affect development of allergic disease or food desensitization. Therefore, massive allergen exposure to genetic skin dysfunction in early infancy and an imbalance of microflora might be necessary for development of atopic march.


Expression of RANKL in osteolytic membranes: association with fibroblastic cell markers.
Aseptic loosening is often mentioned as the primary reason for costly revision of total joint arthroplasties. Receptor activator of nuclear factor-kappaB ligand (RANKL) appears to be a major factor in the bone resorption observed in periprosthetic osteolysis. RANKL plays an essential role in the recruitment, differentiation, and survival of the osteoclasts implicated in periprosthetic osteolysis. This study was performed in an effort to identify the cell type in the periprosthetic membrane responsible for expression of RANKL. Tissues harvested from osteolytic lesions in nine patients undergoing total joint revision were serially sectioned for immunohistochemical analysis. Intercellular adhesion molecule-1 (ICAM-1) and prolyl 4-hydroxylase (5B5) antibodies were used to detect fibroblasts, and anti-CD-163 (Ber-MAC3) was used to detect macrophages. In addition, antibodies to osteoprotegerin (OPG), RANKL, and receptor activator of nuclear factor-kappaB (RANK) were utilized. The binding pattern of these antibodies was then viewed with confocal microscopy with the use of only secondary antibodies as method controls. Histological analysis was confined to areas of the membrane where cells were detected with use of Hoechst 34580 nuclear stain. In the membrane specimens from all nine patients, diffuse RANKL staining was localized to areas lacking cells and more intense staining was seen in areas containing nucleated cells. There was strong colocalization between RANKL and OPG, and there was weak but specific colocalization between RANKL and both 5B5 and ICAM-1. In contrast, there was complete separation of antibody staining of Ber-MAC3 and RANKL, indicating only generalized overlap of the myeloid markers with the RANKL. RANKL expression was localized to cells that stained positively for fibroblast markers. The data also indicated that there is an intact RANKL/RANK/OPG system in the periprosthetic membrane that could regulate focalized bone resorption in osteolysis. Identifying the cell types responsible for RANKL production is critical to the development of a strategy to prevent periprosthetic osteolysis.


Differential interference contrast imaging using a spatial light modulator.
Differential interference contrast microscopy, imaging by interferometric superposition of two displaced beams passed through a transparent sample, is one of the most sophisticated methods in classical microscopy. Here we demonstrate a versatile electronically controlled variant using a liquid-crystal spatial light modulator that displays a diffractive optical element and steers the beam separation. The orientation and magnitude of the shear angle and the relative phase of the two interfering beams can all be varied at video rates. The technique is demonstrated by imaging polystyrene beads in immersion oil and a sample of red blood cells. The method expands the capabilities of previous implementations of differential interference contrast microscopy by its nonmechanical control over all imaging parameters.


The role of vanadium in the management of diabetes.
Diabetes mellitus results from an absolute or relative deficiency in insulin secretion and a resistance of target tissues to the action of insulin, in proportions that vary with the type of the disease. The shortage of insulin can be corrected by administration of exogenous insulin or stimulation of pancreatic beta-cells with sulphonylureas. However, insulin resistance remains a major therapeutic problem. Here, Sonia Brichard and Jean-Claude Henquin review the recent discoveries that indicate a possible role for vanadium in management of the disease. In vitro, vanadium salts mimic most effects of insulin on the main target tissues of the hormone, and in vivo they induce a sustained fall in blood glucose levels in insulin-deficient diabetic rats, and improve glucose homeostasis in obese, insulin-resistant diabetic rodents. Recent short-term clinical trials with vanadium salts also seem promising in type II (non-insulin-dependent) diabetic patients in whom liver and peripheral insulin resistance was attenuated, indicating the therapeutic potential of vanadium salts, pending demonstration of their long-term innocuity.


SAR studies on the central phenyl ring of substituted biphenyl oxazolidinone-potent CETP inhibitors.
SAR studies of the substitution effect on the central phenyl ring of the biphenyl scaffold were carried out using anacetrapib (9a) as the benchmark. The results revealed that the new analogs with substitutions to replace trifluoromethyl (9a) had a significant impact on CETP inhibition in vitro. In fact, analogs with some small groups were as potent or more potent than the CF(3) derivative for CETP inhibition. Five of these new analogs raised HDL-C significantly (>20mg/dL). None of them however was better than anacetrapib in vivo. The synthesis and biological evaluation of these CETP inhibitors are described.


Pralatrexate pharmacology and clinical development.
Folates are well known to be essential for many cellular processes, including cellular proliferation. As a consequence, antifolates, the fraudulent mimics of folic acid, have been shown to be potent therapeutic agents in many cancers. Over the past several decades, efforts to improve on this class of drugs have met with little success. Recently, one analog specifically designed to have high affinity for the reduced folate carrier, which efficiently internalizes natural folates and antifolates, has been shown to be very active in T-cell lymphoma. Pralatrexate, approved by the U.S. Food and Drug Administration in 2009, is highly active across many lymphoid malignancies, including chemotherapy-resistant T-cell lymphoma. Emerging combination studies have now shown that pralatrexate is highly synergistic with gemcitabine, histone deacetylase inhibitors like romidepsin and bortezomib. These insights are leading to a number of novel phase I and II combination studies which could challenge existing regimens like CHOP, and improve the outcome of patients with T-cell lymphoma.


Multiple organ failure during primary HIV infection.
The appearance of primary HIV infection ranges from an asymptomatic presentation to a symptomatic illness resembling infectious mononucleosis. Severe unusual presentations include acute myopericarditis, renal failure, and opportunistic infections such as esophageal candidiasis, cytomegalovirus infection, and Pneumocystis jirovecii pneumonia. We report a case of multiple organ failure during primary HIV infection.


Parent-professional agreement on diagnosis and recommendations for children with developmental disorders.
Twenty-eight children with developmental disorders underwent an interdisciplinary assessment for diagnosis and recommendations about management. Following this assessment, the degree of agreement on diagnosis and recommendations between parents and the team was determined. There was substantial agreement for diagnosis of clearly identifiable conditions such as genetic disorders, autism, and mental retardation but less agreement for milder disorders such as learning disabilities, attention deficit disorder, and emotional disorders. Agreement on management varied, being higher for speech therapy and psychosocial interventions but lower for use of stimulant medication and diet. The parents identified additional diagnostic labels in their child and mentioned more recommendations than the clinicians prescribed. The results suggest the need for careful attention to parent perceptions following interdisciplinary assessment.


Copper deficiency in pregnancy: effect on maternal and fetal polyol metabolites.
The present study was undertaken to determine whether the mortality of the fetus and the neonate of copper-deficient rats consuming fructose during pregnancy is associated with an aberration in carbohydrate metabolism. A total of 84 Sprague-Dawley rats were fed a copper-deficient or a copper-adequate diet containing fructose or starch for 19 or 21 days after conception. The consumption of a fructose-based diet during pregnancy resulted in higher concentrations of maternal blood fructose, sorbitol, triglyceride, and uric acid when compared with a starch diet. The placenta contained more than 10-fold the concentration of glucose and more than double the concentrations of fructose, triglycerides, and sorbitol when fructose was the dietary carbohydrate compared with starch. The livers of fetuses belonging to the fructose dietary group exhibited high concentrations of glucose and sorbitol. In addition, fetal blood contained higher concentrations of glucose, fructose, sorbitol, and triglycerides than the corresponding values from the starch dietary group. The consumption of a copper-deficient diet containing fructose during pregnancy resulted in massive subcutaneous hemorrhages of the fetus. In contrast, this pathology was rare in other dietary groups. The combination of copper deficiency with fructose feeding resulted in more than double the concentration of sorbitol in fetal liver, and higher concentrations of insulin and dopamine of fetal blood compared with the consumption of a copper-deficient diet containing starch.(ABSTRACT TRUNCATED AT 250 WORDS)


Inhibition of 5'-deoxy-5-fluorouridine phosphorolysis by acyclothymidine in tumor cell homogenates.
The inhibitory effect of acyclothymidine[AcyT, 5-methyl-1-(2'-hydroxyethoxymethyl) uracil], a potent pyrimidine nucleoside phosphorylase (PyNPase) inhibitor, on 5'-deoxy-5-fluorouridine (5'-DFUR) phosphorolysis in human and mouse tumor cell homogenates was measured. Competitive inhibition was observed in MKN-74 and Lewis lung carcinoma (LLC), whereas non-competitive inhibition was observed in HeLa. The strength of the inhibitory effect by AcyT showed the following pattern: HeLa < human normal intestine < mouse normal intestine < Colon 26 < LLC < MKN-74 < DLD-1. From the kinetic parameter obtained, we simulated the inhibitory effect of AcyT on 5'-DFUR phosphorolysis in tumor cells and the intestine. These data indicated that AcyT was more sensitive in normal mouse intestine than in Colon 26 and LLC, and that orally administered AcyT can reduce the intestinal toxicity of 5'-DFUR without reducing the antitumor effect in the mouse. The present finding may have an important implication for attempts to introduce AcyT, a potent PyNPase inhibitor, into the clinic.


Ethanol intake-reducing effects of ipsapirone in rats are not due to simple stimulus substitution.
The present series of experiments was conducted to investigate whether the previously reported ethanol intake reducing effects of the 5-HT1A receptor agonist ipsapirone could be based on possible stimulus similarities between both compounds. Rats were trained to discriminate ethanol (12.5% w/v, 1000 mg/kg, IP) from saline in a two-lever food-reinforced drug discrimination (DD) procedure. Discrimination criterion was reached after a mean number of training sessions of 42. In subsequent generalization sessions, a dose-response curve was established for ethanol (125-1000 mg/kg, IP, ED50 value: 355 mg/kg). In additional cross-generalization tests with ipsapirone (1-30 mg/kg, IP), stimulus substitution for the ethanol cue was not noted (maximal degree of generalization: 33%, at 10 and 30 mg/kg). To confirm the DD findings that ipsapirone does not substitute for ethanol, an alternative cross-familiarization conditioned taste aversion paradigm (CF-CTA) was utilized. In rats, 1000 mg/kg IP ethanol was used as the reference drug producing a conditioned taste aversion (CTA). It was found that preexposure to ethanol (500-1500 mg/kg, IP) dose-dependently attenuates the CTA produced by this same drug. Full familiarization was noted with 1000 and 1500 mg/kg. In contrast with this, ipsapirone (1-30 mg/kg, IP) failed to abolish ethanol-induced CTA, suggesting again that the ipsapirone stimulus complex is dissimilar to that produced by ethanol. Because the present findings indicate that, in rats, ipsapirone does not substitute for ethanol, it is suggested that the reported ethanol intake-reducing effects of ipsapirone in animal models of alcoholism are not due to simple stimulus substitution.


Histomorphology and microradiography of quadriceps tendon-patellar bone grafts in press-fit anterior cruciate ligament reconstruction.
The press-fit fixation technique has become an alternative fixation method in anterior cruciate ligament (ACL) reconstruction. While the stability of press-fit fixated grafts reported in the literature predominantly had been analysed by biomechanical testing, this study investigated the specific histomorphologic and microradiographic features important for graft stability, using human quadriceps tendon-patellar bone (QTPB) grafts. Six QTPB grafts with cylindrical bone plugs were obtained from frozen fresh human cadaver knees and fixed to porcine, bovine, or human femora in a press-fit technique. The specimens were plastinated, dissected into 1.5 mm slices, studied microradiographically and stained for histologic evaluation. The specimens showed an osseous inclusion of the graft within the femoral tunnel, which was most evident on the cancellous side. Histologic analysis revealed a trabecular interdigitation with a penetration of patellar and femoral trabecular structures, contributing to primary stability of the press-fit fixation. Graft inclusion was most obvious in bovine and least in human femora, assumably because of varying bone quality. The circular bone-to-bone contact in press-fit fixation technique should contribute to both rapid and stable graft healing.


Childhood steroid-sensitive nephrotic syndrome: does the histology matter?
Renal biopsy specimens from 51 children with steroid-sensitive nephrotic syndrome who were following a frequently relapsing or steroid-dependent course were reviewed by two histopathologists. In all cases the biopsy was performed prior to the commencement of an 8-week course of cyclophosphamide. The clinical courses of these patients both prebiopsy and for a minimum of 2 years after completion of cyclophosphamide therapy were analyzed using retrospective case note analyses. The distribution of histologic diagnoses differed significantly from that reported by the International Study of Kidney Disease in Children in their study of children who underwent biopsy at the time of presentation, there being an excess of focal segmental glomerulosclerosis and mild mesangial hypercellularity in this series. The prebiopsy clinical course did not predict for histologic diagnosis, and there was no correlation between prebiopsy course or histology and postcyclophosphamide course. The findings of this study support the clinical impression that steroid sensitivity rather than histology is the major determinant of prognosis in childhood nephrotic syndrome and that frequency of relapse alone is not an indication for biopsy.


Anticonvulsant effect of unilateral anterior thalamic high frequency electrical stimulation on amygdala-kindled seizures in rat.
Deep brain stimulation (DBS) is an emerging treatment of epilepsy. Anterior nucleus of the thalamus (ANT) is considered to be an attractive target due to its close connection to the limbic structures and wide regions of neocortex. In this study, we examined the effect of unilateral high frequency stimulation (HFS) of the ANT on amygdala-kindled seizures in Wistar rats. When fully-kindled seizures were achieved by daily amygdala kindling, HFS (15 min train of 100 μs pulses at 200 Hz and 450-800 μA) was delivered to the ipsilateral or contralateral ANT immediately before the kindling stimulation for 15 days. HFS of the ipsilateral ANT significantly decreased the incidence of generalized seizures and the mean behavioral seizure stage and afterdischarge duration (ADD), and shortened cumulative ADD and cumulative generalized seizure duration. Furthermore, HFS of the ipsilateral ANT significantly increased the afterdischarge threshold (ADT). Our data suggest that unilateral HFS of the ANT may be an effective method of inhibiting kindled seizures by suppressing the susceptibility to seizures and generating long lasting anti-epileptic effect preventing the recurrence of kindled seizures, providing an alternative to bilateral ANT DBS for refractory epilepsy.


Herpetic Brachial Plexopathy: Application of Brachial Plexus Magnetic Resonance Imaging and Ultrasound-Guided Corticosteroid Injection.
Herpes zoster, commonly known as shingles, is an infectious viral disease characterized by painful, unilateral skin blisters occurring in specific sensory dermatomes. Motor paresis is reported in 0.5% to 5% of patients. Although the mechanism of zoster paresis is still unclear, the virus can spread from the dorsal root ganglia to the anterior horn cell or anterior spinal nerve roots. It rarely involves the brachial plexus. We report a case of brachial plexitis following herpes zoster infection in which pathological lesions were diagnosed using brachial plexus magnetic resonance imaging and treated with ultrasound-guided perineural corticosteroid injection.


Cognate sensor kinase-independent activation of Mycobacterium tuberculosis response regulator DevR (DosR) by acetyl phosphate: implications in anti-mycobacterial drug design.
The DevRS/DosT two-component system is essential for mycobacterial survival under hypoxia, a prevailing stress within granulomas. DevR (also known as DosR) is activated by an inducing stimulus, such as hypoxia, through conventional phosphorylation by its cognate sensor kinases, DevS (also known as DosS) and DosT. Here, we show that the DevR regulon is activated by acetyl phosphate under 'non-inducing' aerobic conditions when Mycobacterium tuberculosis devS and dosT double deletion strain is cultured on acetate. Overexpression of phosphotransacetylase caused a perturbation of the acetate kinase-phosphotransacetylase pathway, a decrease in the concentration of acetyl phosphate and dampened the aerobic induction response in acetate-grown bacteria. The operation of two pathways of DevR activation, one through sensor kinases and the other by acetyl phosphate, was established by an analysis of wild-type DevS and phosphorylation-defective DevSH395Q mutant strains under conditions partially mimicking a granulomatous-like environment of acetate and hypoxia. Our findings reveal that DevR can be phosphorylated in vivo by acetyl phosphate. Importantly, we demonstrate that acetyl phosphate-dependent phosphorylation can occur in the absence of DevR's cognate kinases. Based on our findings, we conclude that anti-mycobacterial therapy should be targeted to DevR itself and not to DevS/DosT kinases.


Isolation and genetic characterization of avian origin H9N2 influenza viruses from pigs in China.
As pigs are susceptible to infection with both avian and human influenza A viruses, they have been proposed to be an intermediate host for the adaptation of avian influenza viruses to humans. In April 2006, a disease caused by highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) occurred in several pig farms and subsequently overwhelmed almost half of China with more than 2,000,000 cases of pig infection. Here we report a case in which four swine H9N2 influenza viruses were isolated from pigs infected by highly pathogenic PRRSVs in Guangxi province in China. All the eight gene segments of the four swine H9N2 viruses are highly homologous to A/Pigeon/Nanchang/2-0461/00 (H9N2) or A/Wild Duck/Nanchang/2-0480/00 (H9N2). Phylogenetic analyses of eight genes show that the swine H9N2 influenza viruses are of avian origin and may be the descendants of A/Duck/Hong Kong/Y280/97-like viruses. Molecular analysis of the HA gene indicates that our H9N2 isolates might have high-affinity binding to the alpha2,6-NeuAcGal receptor found in human cells. In conclusion, our finding provides further evidence about the interspecies transmission of avian influenza viruses to pigs and emphasizes the importance of reinforcing swine influenza virus (SIV) surveillance, especially after the emergence of highly pathogenic PRRSVs in pigs in China.


[99mTc-MDP abnormal uptake in the femur shaft in hemi-lateral hip joint disorders].
During the past 4 years, three-phase bone scintigraphy using 99mTc-MDP has been studied in 68 patients suffering from hemi-lateral hip-joint disorders. We were impressed on abnormal uptake of the shaft of femur on the involved side. 99mTc-MDP uptake in the shaft of femur is compared with between involved leg and normal leg of 68 cases with hemi-lateral hip-joint disorders (76 examinations). We excluded cases of osteomyelitis, cases of malignant tumor, and post-operative cases. In early images, the 99mTc-MDP uptake is not always increased in involved side. But in delayed images, there are no patients whose normal side's 99mTc-MDP uptake is increased. The 99mTc-MDP uptake in early image is not related only with blood flow of the thigh but with the amount of soft tissue of the thigh. And the 99mTc-MDP uptake in delayed image is related with disuse osteoporosis.


Hematoidin crystals in cervicovaginal smears. Report of two cases.
Cervicovaginal smears obtained from two women, one at the 32nd week of pregnancy and the other at the immediate postpartum period, contained hematoidin crystals. These crystals were of a golden color and appeared as radially arranged fine needles forming cockleburrs or as aggregates of small irregular particles, spherules or rhomboids, in most instances associated with or within histiocytes. Hemosiderin pigment was associated with them in small amounts. The presence of hematoidin crystals is not an uncommon finding in cytologic specimens, particularly those obtained by fine needle aspiration of walled-off, necrotizing lesions with a component of hemorrhage. It is a very unusual finding in cervicovaginal smears, probably because of the ability of the uterus to promptly rid itself of blood. Certain aspects of the formation and identity of hematoidin are discussed.


[Clinical role of evaluating vascular paralysis in septic shock patients].
To explore the effects of vascular paralysis upon prognosis and tissue perfusion in septic shock patients. A total of 73 septic shock patients consecutively admitted into our department from January 2010 to July 2011 were retrospectively studied. Their hemodynamic data, arterial lactate concentration and APACHEII (acute physiology & chronic health evaluation II) score at the beginning of cardiac output monitoring (0h), as well as the outcome of Day 28 post-diagnosis were recorded. Stroke volume index (SVI) and cardiac index (CI) were obtained through transpulmonary thermodilution technique by pulse induced continuous cardiac output (PiCCO) system. (1) APACHEII score at 0h was higher in non-survivors than those in survivors ((14.6 ± 5.6) vs (20.4 ± 4.8), P < 0.01). Stroke systemic vascular resistance index (SSVRI), pulse pressure/stroke volume index (PP/SVI), effective arterial elastance index (EaI), 24h lactate clearance rate (24rLac) , 72h lactate clearance rate (72rLac) and 7d lactate clearance rate (7rLac) in non-survivors were all lower than those in survivors (P < 0.05); (2) SVRI, SSVRI, PP/SVI and EaI were all correlated significantly with 24rLac, 72rLac and 7rLac [ (212.7 ± 52.6) vs (185.4 ± 50.5) , PP/SVI: (2.8 ± 0.7) vs (2.5 ± 0.6), EaI: (5.0 ± 1.2) vs (4.3 ± 1.1), 24/72/7 rLac: (18.4 ± 46.4) vs (21.5 ± 49.7), (19.9 ± 49.6) vs (-21.5 ± 46.3), (35.5 ± 45.8) vs (-59.5 ± 64.5), P < 0.00]. Vascular paralysis is correlated with prognosis and tissue perfusion in septic shock patients. And SSVRI, PP/SVI and EaI are more significant.


Zinc homeostasis in aging: two elusive faces of the same "metal".
Proteins involved in zinc homeostasis may be altered in aging. This phenomenon may lead to zinc deficiency in the peripheral blood and an accumulation of zinc bound to insoluble aggregates at the extracellular level in the brain. Therefore, it should be more correct to talk about aging as a condition associated with zinc dyshomeostasis rather than deficiency. Restoring functional zinc homeostasis in aging people is an attractive field for antiaging research, but requires further knowledge than the current state of the art.


The dischargeable cut-off score of Oswestry Disability Index (ODI) in the inpatient care for low back pain with disability.
The admission due to low back pain (LBP) became prevalent cause of international economic losses. Since LBP patients with disability are often subject to inpatient care, it is important to determine the appropriate time of discharge. The purpose of this study is to set the cut-off value of appropriate Oswestry Disability Index (ODI) at the time of discharge. Of 1,394 LBP patients admitted in hospital specialized in spinal disease, 774 eligible patients with disability were included in this study. And several clinical variables including numerical rating score, ODI, satisfaction level were observed during the hospital stay. We considered satisfaction level as an important factor for discharge, categorized patients into satisfied group and dissatisfied group. Through the statistical analysis, appropriate factor for determining dischargeable patients satisfied with their current condition and its cut-off value of ODI were found. And proper predictors for the cut-off value were extracted statistically and logically from a pool of several clinical indexes. The ODI at the time of discharge was most accurate in determining dischargeable patients. The cut-off value of ODI was 30. Predictors were ODI questions 4 and 6. We set the cut-off value of dischargeable ODI for LBP inpatient with disability and found its predictor.


[Immunohistochemical localization of estrogen receptors in vesicovaginal fascia among female patients with stress incontinence--preliminary report].
Using monoclonal antibody assay technique 29 kD domain of estrogen receptor was found and partially quantitated by means of histological score in fibroblasts from vesico-vaginal fascia obtained during surgical procedure in incontinent women. This is an important proof that estrogen supplementation therapy is a reasonable approach in women affected by urinary stress incontinence.


Effect of surfactants on weight gain in mice.
A study was conducted to determine if four surfactants can induce increased weight gain in the mouse. Basic-H, Triton X-100, Amway All Purpose Adjuvant and X-77 were put in water and fed to various groups of ICR 21 day old female mice for a period of 43 days. All the mice were clinically normal throughout the study period. Pathological examination of a random sample of the mice revealed no gross pathological changes. Similarly, histopathological examination of the lungs, livers and intestines did not reveal any visible lesions. Basic-H and Amway surfactants induced weight gain, though not significantly, better at 0.1% (V/V) concentration while X-77 and Triton X-100 induced weight gain better at 0.4% (V/V) concentration. Overall results show that none of the surfactants tested induced significant weight gain.


Genotypic and technological diversity of Brevibacterium linens strains for use as adjunct starter cultures in 'Pecorino di Filiano' cheese ripened in two different environments.
Twenty-two Brevibacterium linens strains isolated from 'Pecorino di Filiano' cheese ripened in two different environments (natural cave and storeroom) were characterized and differentiated for features of technological interest and by genotypic methods, in order to select strains with specific features to be used as surface starter cultures. Results showed significant differences among strains on the basis of physiological and technological features, indicating heterogeneity within the species. A middle-low level of proteolytic activity was observed in 27.3 % of strains, while a small group (9.1 %) showed a high ability. Lipolytic activity was observed at three different temperatures and the highest value was detected at 20 °C with 13.6 % of strains, while an increase in temperature produced a slightly lower lipolysis in all strains. The evaluation of diacetyl production revealed that only 22.8 % of strains showed this ability, and most of them were isolated from product ripened in the natural cave. All strains exhibited only leu-aminopeptidase activity, with values more elevated in strains coming from the natural cave product. The combined analysis of genotypic results with the data obtained by the features of technological interest study established that the random amplified polymorphic DNA (RAPD) clusters obtained were composed not only of different genotypes but of different profiles based on technological properties too. This study demonstrated the importance of the ripening environment that affects the typical features of the artisanal product, leading to the selection of a specific surface microflora. Characterized strains could be associated within surface starters to standardize the production process of cheese, but preserving its typical organoleptic and sensory characteristics and improving the quality of the final product.


The relation of Cotard's syndrome to delusional misidentification.
A case of Cotard's syndrome with face processing deficits is described. The phenomenology of Cotard's syndrome may be an expression of underlying visual processing deficits, including abnormalities in face perception processing. Phenomenologic and neurobiologic characteristics of this case are similar to those found among delusional misidentification syndromes. This similarity suggests areas for further study that may lead to a better understanding of the relation of phenomenology to neurobiology in Cotard's syndrome.


[Listeria infections and pregnancy].
One clinical case of lysteriosis during pregnancy, diagnosed in the puerperium period is analyzed. Both the mother and the newborn recovered favorably.


Bayclone: Bayesian nonparametric inference of tumor subclones using NGS data.
In this paper, we present a novel feature allocation model to describe tumor heterogeneity (TH) using next-generation sequencing (NGS) data. Taking a Bayesian approach, we extend the Indian buffet process (IBP) to define a class of nonparametric models, the categorical IBP (cIBP). A cIBP takes categorical values to denote homozygous or heterozygous genotypes at each SNV. We define a subclone as a vector of these categorical values, each corresponding to an SNV. Instead of partitioning somatic mutations into non-overlapping clusters with similar cellular prevalences, we took a different approach using feature allocation. Importantly, we do not assume somatic mutations with similar cellular prevalence must be from the same subclone and allow overlapping mutations shared across subclones. We argue that this is closer to the underlying theory of phylogenetic clonal expansion, as somatic mutations occurred in parent subclones should be shared across the parent and child subclones. Bayesian inference yields posterior probabilities of the number, genotypes, and proportions of subclones in a tumor sample, thereby providing point estimates as well as variabilities of the estimates for each subclone. We report results on both simulated and real data. BayClone is available at http://health.bsd.uchicago.edu/yji/soft.html.


Biosynthesis and secretion of adrenomedullin and proadrenomedullin N-terminal 20 peptide in a rat model of endotoxin shock.
To study the biosyntheses and pathophysiological roles of adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) in septic shock, we compared the time course of plasma concentrations of these peptides and blood pressure in rats injected with either 0.9% saline (control group) or lipopolysaccharide (LPS group). The plasma AM concentration in the LPS group did not increase 30 and 60 min after LPS injection, at which time points the blood pressure remained low. Thereafter, AM rapidly increased, and it amounted to 35 times the basal value 4 h after injection, when the blood pressure returned to the basal level. The increment of plasma PAMP in the LPS group was lower than that of AM. We also examined the tissue concentration of AM and PAMP--as well as the tissue expression of proadrenomedullin (proAM) mRNA--in the LPS and control groups. LPS significantly increased the tissue concentrations of AM and PAMP in the lung, but decreased them in the adrenal gland and cardiac atrium. The LPS injection augmented proAM gene transcription in the lung, adrenal gland and aorta. In an immunohistochemical examination, AM staining was intense in alveolar endothelial cells of the lung in the LPS group. Thus, this septic shock model had high plasma levels of PAMP as well as AM, while the biosynthesis and secretion of the two peptides may have been differentially regulated in various tissues of rats injected with LPS. The present results suggest that these two bioactive peptides may play different roles in the pathophysiology of septic shock.


Wound dressing in major head and neck cancer surgery: a prospective randomized study of gauze dressing vs sterile vaseline ointment.
A total of 207 patients were randomized in a prospective comparative study of standard gauze dressing vs sterile vaseline ointment. 179 patients were evaluable. All patients received antimicrobial prophylaxis. The two groups (86 standard and 93 vaseline) were comparable as far as age (mean, 57 yr; range, 21-84), genders (155 males/24 females), weight (mean, 66 kg; range, 40-69), type of surgery, previous or concomitant anticancer treatment. Severity of surgery was identical, as was the severity of cancer, in the two groups. Wound infection within 20 days of surgery occurred in 31.2% (29/93) of the vaseline group and 24.4% (21/86) in the standard group (NSS). Bacteremia occurred in three patients from the vaseline group and in four patients from the standard group. Bronchopneumonia occurred in 10 patients from the vaseline group and 14 patients in the standard group. The spectrum of microorganisms recovered was similar in the two groups. The need for antimicrobial treatment (empiric or for documented infections) within 20 days after surgery was 34.4% (32/93) in the vaseline group and 36.0% (31/86) in the standard group. The median delay to infection (range in days) in the vaseline group was 9 (5-15) for wound and 6 (1-12) for bronchopneumonia. For the standard group the corresponding delays were 8 (4-15) and 7 (2-19). Vaseline dressing was not associated with an increased risk of infection as compared to the standard gauze dressing.


Genetic mapping and physical cloning of UVB susceptibility region in mice.
One of the most important goals of cancer research is to identify environmental and host factors that contribute to the malignant state. Human skin cancers are among the few tumor types for which the predominant environmental carcinogen is known. Ultraviolet light, a component of sunlight, is an important cause of skin cancer in humans. In humans and mice, ultraviolet B radiation induces systematic and local immunosuppression. A consequence of that is inappropriate immune surveillance of somatic tissues for evidence of malignantly transformed cells. The impairment of contact hypersensitivity, as it develops early and correlates well with tumor frequency in various mouse strains, has been used for over 15 y as a model of immunologic events occurring in photocarcinogenesis. In mice, as well as in humans, ultraviolet B radiation induced impairment of contact hypersensitivity is not uniform in all individuals; some individuals are susceptible to the deleterious effects of ultraviolet B, whereas others are resistant to ultraviolet B. We have defined the genetic locus responsible for ultraviolet B susceptibility and resistance in mice within the Bat5 and H-2D segment of the mouse chromosome 17.


Interaction of gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) with DNA.
The size and structure of aggregates formed by interaction of DNA with homologous series of cationic gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) (CnGS, n=10-16 is the number of alkyl carbons) were investigated using UV-vis turbidity, dynamic light scattering and small-angle synchrotron X-ray (SAX) diffraction. The detailed analysis of turbidity in the range of lambda=450-600 nm indicates an anomaly in the growth of CnGS+DNA aggregates with increasing concentration of CnGS, possibly involving changes of structure and size of aggregates. Using dynamic light scattering, changes of the effective diameter of CnGS+DNA (n=12 and 16) aggregates formed in the CnGS concentration range 0.002-0.140 mmol/l were observed. SAX diffractograms show the presence of long-range organization of CnGS+DNA (n=12, 13, 14 and 16) aggregates due to DNA interaction with CnGS above the critical micellar concentration. The CnGS+DNA (n=12, 13 and 14) aggregates at 25 degrees C are packed in a lattice of two-dimensional hexagonal symmetry. With increasing C14GS:DNA molar ratio the changes of the lattice parameter in the range of 4.80-5.27 nm are observed at 25 degrees C. The aggregates undergo structural changes induced by temperature in the range 60-95 degrees C, which are accompanied by changes of the diffraction patterns, namely in the region of reciprocal spacing s=0.15-0.30 nm(-1).


Novel properties of P450s in Streptomyces coelicolor.
In the process of investigating the function of the 18 P450s that make up the CYPome in the soil bacterium Streptomyces coelicolor, we have discovered new and novel biochemical features of some of these monooxygenases. This article which is dedicated to Ron Estabrook summarizes these discoveries with emphasis on their novelty. It is concluded from this work that there are many interesting features of P450 enzymes waiting to be discovered. Since there are more than 6500 P450 gene sequences known today, we can expect that many new biochemical aspects of P450s will be discovered as these individual monooxygenases are investigated.


Cloning and characterization of cholesteryl ester transfer transfer protein isolated from the tree shrew.
To obtain the nucleotide sequence and deduced amino acid sequence of cholesteryl ester transfer protein (CETP) cDNA from the tree shrew (Tupaia glis). The cDNA sequence of the tree shrew CETP was obtained by utilizing the technique of switching mechanism at 5' end of RNA transcript (SMART) and rapid amplification of cDNA end (RACE) from the first strand of the cDNA. The amino acid sequence of CETP was deduced from the cDNA sequence and its primary and secondary structures were predicted. The sequence of CETP cDNA from tree shrew (GenBank accession number AF334033) covers 1636 bp, including 178 bp at the 3' end of the untranslated region and a 1458 bp fragment in a coding region, which provides the complete sequence of mature tree shrew CETP, although not the initiator methionine. The first 24 bp encodes a partial signal peptide. The mature protein consists of 477 amino acids and is longer than the human version by one amino acid (Gly318). Comparing this amino acid sequence with those of other animals' CETPs, the identity between tree shrew and human and rabbit CETP is 88% and 82%, respectively. The protein is extremely hydrophobic as it contains many hydrophobic residues, especially at the C-terminal, consistent with its function in the transfer of neutral lipids. The amino acid residues concerning with binding and transferring neutral lipids are highly conserved. There is a deletion of an N-linked glycosylation site at Asn342 in the tree shrew CETP protein that may participate in the removal of peripheral cholesterol and cholesteryl ester by increasing its activity of transferring cholesteryl ester. The possible glycosylation in the tree shrew CETP may be involved in the molecular mechanism of its insusceptibility to atherosclerosis.


Induction of G2/M arrest and apoptosis in human gastric epithelial AGS cells by aqueous extract of Agaricus blazei.
Agaricus blazei is well known as a traditional medicinal mushroom and it has been shown to exhibit immunostimulatory and anti-cancer activity. However, the cellular and molecular mechanism of cell cycle arrest and apoptosis of cancer cells is poorly understood. In this study, we have investigated whether A. blazei extract (ABE) exerts anti-proliferative and apoptotic effects on human gastric epithelial AGS cells. It was found that ABE could inhibit cell growth in a dose-dependent manner, which was associated with the arrest of G2/M phase and the induction of apoptotic cell death. Flow cytometric analysis showed that ABE could cause an arrest at the G2/M phase of the cell cycle, which is closely correlated to decreased cyclin B1 and cdc2 levels. Furthermore, this compound induced apoptosis through up-regulation of Bax and the activation of caspases with down-regulation of XIAP and cIAP-1, but not cIAP-2, and a capase-3 inhibitor could block cell death and apoptotic body formation. These data clearly indicate that ABE-induced apoptosis is associated with caspase-3 activation. In summary, the growth inhibition of ABE is highly related to cell cycle arrest at the G2/M phase and the induction of caspase-3-dependent apoptosis in human gastric epithelial AGS cells.


Problems in the determination of prostate specific antigen.
Accurate determination of prostate specific antigen is important because of the increasing use of prostate specific antigen for diagnosis, follow-up and screening of prostate cancer. Standardization of this assay is complicated by the occurrence in serum of two major molecular forms of prostate specific antigen, free prostate specific antigen and a complex between prostate specific antigen and alpha 1-antichymotrypsin. These two forms of prostate specific antigen are recognized differently by many but not all antibodies. Thus, it is possible and desirable to develop methods that measure each form equally. To achieve better comparability, it is also necessary to prepare international standards for prostate specific antigen and its complex with alpha 1-antichymotrypsin. Furthermore, reference methods should be established which use these standards and carefully selected reference antibodies.