[Lung embolism during and following long-distance flights ("economy class syndrome")].
Two cases of venous thrombosis and pulmonary embolism during or after intercontinental air travel occurring in two sisters with venous thromboembolic risk factors are presented. In one the outcome was fatal. In the present era of mass tourism the probably underestimated risk of thromboembolism in air travel should be kept in mind. As prophylactic measures leg exercises, walking and high fluid intake during the flight are recommended. Sleeping in the sitting position should be avoided. For persons at high risk, prophylactic anticoagulation may be considered.


Mood patterns and variations associated with personality disorder pathology.
This study examined mood and mood variation in relation to varying forms and degrees of personality disorder (PD) pathology. Mood experiences of 98 psychotropic medication-free individuals were repeatedly assessed over a 4-day period. Persons with PDs (n = 57) generally displayed neutral to moderately positive moods; however, overall mood valence was less positive when compared to those without PDs (n = 41). Mood ratings demonstrated moderate covariations with anxious-fearful (A-F) PD traits but little or no association with erratic-emotional-dramatic (E-D) and odd-eccentric (O-E) PD traits once common variance among PD dimensions was removed. For PD diagnostic categories, the presence of avoidant and/or depressive PDs was most strongly associated with negative mood. When dimensional scores based on specific PD trait features were considered, avoidant, depressive, borderline, passive-aggressive, obsessive-compulsive, dependent, paranoid, and schizoid PD traits demonstrated the most reliable associations with negative mood. Apart from borderline PD features, traits associated with other E-D cluster PDs displayed little or no associations with mood quality. Consistent with previous research, mood variability emerged as an internally consistent and stable individual difference variable. Mood variability, however, was not generally associated with PD diagnostic categories or traits. Implications of this study's findings are considered in relation to the conceptual modeling of PDs.


Safety evaluation of magnetic catheter steering with upgraded magnetic resonance imaging system.
Catheter navigation and placement through the arterial network is a major limitation for clinical procedure. In this article, a specific catheter tip and a modified clinical MRI scanner with an upgraded gradient system are used to steer a catheter through a single Y-shaped bifurcation. Safety aspects are analyzed to avoid the peripheral nerve stimulation (PNS) according to an empirical law of magnetostimulation and the magnetic field of upgraded 3D gradient coils. For a rabbit-sized device, the rising time of gradients system have to be limited to 1.7ms at 400mT.m(-1) to avoid PNS. These rise time values allow the use of this system for catheter steering and other more demanding applications.


Tumor-promoting effect of GGN-MRP extract from the Maillard reaction products of glucose and glycine in the presence of sodium nitrite in C3H10T1/2 cells.
GGN-MRP is an extract from the Maillard reaction products of nitrite with glucose and glycine in the Maillard browning system. No genotoxicity of GGN-MRP in culture hepatocyte was found. A two-stage transformation protocol was used to transform chemically mouse embryo fibroblast C3H10T1/2 cells. To initiate transformation, the cells were treated with benzo[a]pyrene [B(a)P; 0.1 microg/mL], and GGN-MRP (0.01, 0.1, and 1.0 mg/mL) was employed to subsequently complete the transformation process. Malignant transformed foci were formed in B(a)P-initiated and GGN-MRP-promoted C3H10T1/2 cells after 8 weeks. Cells treated with GGN-MRP alone failed to induce transformation. However, cells initiated with B(a)P and promoted by GGN-MRP demonstrated oncogenic properties. Transformed colonies derived from GGN-MRP-treated cells exhibited enhanced growth rate, anchorage independence, and tumorgenicity in animals relative to parent cells. These results indicated that GGN-MRP contains a tumor promoter and may induce tumor promotion by two-stage oncogenesis.


Information-seeking behaviors and response to the H1N1 outbreak in Chinese limited-English proficient individuals living in King County, Washington.
To investigate the information seeking behaviors and response to the HIN1 outbreak by limited-English proficient (LEP) Chinese speakers. We conveniently sampled 100 adult Chinese LEP individuals between June 2 and June 11, 2009, during the time the World Health Organization (WHO) declared global pandemic alert level at phase 6 and the development of a H1N1 vaccine was still underway. Participants demonstrated a basic understanding of the disease and were unconcerned by the outbreak. Major channels for HIN1 information included watching TV (81 percent), reading Chinese newspaper (69 percent), and community-based organization (30 percent). Only 2 percent obtained information from a public health system or hotline. The odds of being informed of timely HIN1 information were significantly higher for participants who did not speak English at all than those who reported speaking English "not well"(OR = 2.65; CI:1.04, 7.01). LEP Chinese speakers seem acknowledged for this outbreak. However, scarce use of the local public health system to obtain H1N1 information suggests more work needs to be done to reach out to the LEP community to enhance their capacity to respond to future outbreaks.


Humoral factors of natural resistance of Bos indicus cattle selected for antibody titre to Brucella abortus.
The concentration of lysozyme, total immunoglobulin and bactericidal activity were measured in sera of Bos indicus cattle, retrospectively screened for specific antibodies to Brucella abortus and classified as being positive reactors or negative reactors. In addition, the effect of complement in the sera was studied to demonstrate complement dependence of antibody-mediated bacterial killing. It was observed that, under the test conditions, serum bactericidal activity and concentration of total immunoglobulin were associated with high specific antibody levels (P less than 0.001). Furthermore, there was a slight decrease in the lytic activity of lysozyme in the sera of animals with high antibody titres.


The electrophysiology of tactile extinction: ERP correlates of unconscious somatosensory processing.
We examined the electrophysiological correlates of left-sided tactile extinction in a patient with right-hemisphere damage. Computer-controlled punctate touch was presented to the left, right or both index fingers in an unpredictable sequence. The patient reported his conscious tactile percept ("left", "right" or "both"). He showed extinction on 75% of bilateral trials, reporting only right stimulation for these. Somatosensory evoked potentials for unilateral stimulation showed early components over contralateral somatosensory areas (P60 and N110) for either hand. In contrast to the results observed for age-matched controls, the patient's P60 was smaller in amplitude for left-hand touch over the right hemisphere than for right-hand touch over the intact hemisphere. Bilateral trials with extinction revealed residual P60 and N110 components over the right hemisphere in response to the extinguished left touch. These results demonstrate residual unconscious somatosensory processing of extinguished touch. They also suggest that tactile extinction can be caused by attenuation rather than elimination of somatosensory responses in the damaged hemisphere, with an underlying deficit even on unilateral trials.


E-1020, a water soluble imidazopyridine, has direct effects on Ca(2+)-dependent force and ATP hydrolysis of canine and bovine cardiac myofilaments.
E-1020 is a cardiotonic agent that acts as a cyclic-AMP phosphodiesterase inhibitor but also may have actions which alter myofilament response to Ca2+. To identify direct actions of E-1020 on cardiac contractile proteins, effects of E-1020 on myofibrillar Ca2+ dependent MgATPase and force generation in chemically skinned fiber bundles were measured. In bovine cardiac myofibrils, E-1020 (100 microM) significantly increased myofilament Ca2+ sensitivity and Ca(2+)-dependent ATPase activity at submaximal pCa values. At pCa 6.75, E-1020 significantly increased ATPase activity in bovine (10-100 microM) and canine (1-100 microM) cardiac myofibrils but had no effect on rat cardiac myofibrils. Moreover, in one population of canine ventricular fiber bundles, E-1020 (0.01-10 microM) significantly increased isometric tension at pCa 6.5 and 6.0, whereas in another population of bundles E-1020 had no effect on tension. In no case was resting (pCa 8.0) or maximal tension (pCa 4.5) increased by E-1020. Measurements of Ca2+ binding to canine ventricular skinned fiber preparations demonstrated that E-1020 does not alter the affinity of myofilament troponin C for Ca2+. We conclude that part of the mechanism by which E-1020 acts as an inotropic agent may involve alterations in the responsiveness of contractile proteins to Ca2+. The lack of effect of E-1020 on some preparations may be dependent on isoform populations of myofilament proteins.


Mothers' intention, age, education and the duration and management of breastfeeding.
The authors examined the breastfeeding duration and management of two groups of mothers with different exposures to services of a Certified Lactation Consultant (CLC). One group of mothers, at hospital H1 (n = 46), had access to a CLC, while mothers at hospital H2 (n = 115) did not. Results showed that: (a) mothers at H1 had significantly (t = 2.33, p < .02) longer durations of breastfeeding (M = 3.1 months, SD = 1.2) than peers at H2 (M = 2.4 months, SD = 1.2); (b) a significantly greater proportion of mothers at H1 attained their intended duration of breastfeeding compared to mothers at H2 (Mann-Whitney U, one-tailed test, Z = 1.94, p < .05); and (c) in a stepwise multiple regression analysis, intended length of breastfeeding accounted for 18% of the variance in duration of breastfeeding, mothers' age 9%, and mothers' education 3%. The results support the theory of reasoned action and the theory of patient education.


Relationships between digit ratio (2D:4D) and basketball performance in Australian men.
To investigate relationships between the digit ratio (2D:4D) and competitive basketball performance in Australian men. Using an observational cross-sectional design a total of 221 Australian basketball players who competed in the Olympic Games, International Basketball Federation World Championships/Cup, Australian National Basketball League, Central Australian Basketball League or socially had their 2D:4Ds measured. Analysis of variance was used to assess differences in mean 2D:4Ds between men playing at different competitive standards, with relationships between 2D:4Ds and basketball game-related statistics assessed using Pearson's product moment correlations in men playing at a single competitive standard. There were significant differences between competitive standards for the left 2D:4D following Bonferroni correction, but not for the right 2D:4D, with basketballers who achieved higher competitive standards tending to have lower left 2D:4Ds. No important correlations between 2D:4D and basketball game-related statistics were found, with correlations typically negligible. This study indicated that the 2D:4D can discriminate between basketballers competing at different standards, but not between basketballers within a single competitive standard using objective game-related statistics.


[Immunglobulin levels after kidney transplantation and during rejection crisis (author's transl)].
In the first three weeks after kidney transplantation the patients were examined for daily changes in immunoglobulin levels. Only very seldom was a decrease of IgM detected in our patients who had undergone splenectomy. During half of the rejection crisis there were indeed short periods of increase (spikes) in immunoglobulins. These were observed always before or concomitantly with the clinical diagnosis of a rejection reaction. In order to determine the connection between immunoglobulin-spikes and an approaching rejection episode the general one-sided binominal test was used. A significant relation was established between IgM peaks and an approaching rejection crisis. No such relation could be found for the IgA peaks observed. The IgG globulins showed no consistent pattern during a transplantation rejection. A persistent fall of the IgG globulins was noted in the first ten days after renal allograft with the immunosuppressive regimen of an initial gramme of methylprednisolon and high dosage of azothioprine. A good correlation was observed with the simultaneous depression of serum albumin concentrations. As losses of protein in urine are minimal (already confirmed by other authors), it must be assumed that in the first place a high rate of catabolism and a restriction in the protein synthesis is responsible for the decrease in the IgG globulins and the serum albumins.


The uptake of the trypanocidal drug suramin in combination with low-density lipoproteins by Trypanosoma brucei and its possible mode of action.
In plasma, a significant part of suramin circulates in tight association with low-density lipoproteins (LDL). At therapeutically obtainable concentrations (100 microM) of suramin, about 85% of the total amount of the drug was bound to proteins, approximately 15% of which was bound to LDL. The molar ratio of suramin bound to LDL in serum was 7.5. The capacity of the high-affinity binding sites of LDL were 6.6 x 10(6) M-1, both in Tris buffer and in ultrafiltrate of serum. Suramin (100 microM) decreased the uptake of host LDL through receptor-mediated endocytosis by Trypanosoma brucei, with approximately 50%. LDL served as the only carrier for suramin uptake. Serum albumin, another important carrier for suramin in blood, was not able to promote suramin uptake, neither was delipidified plasma. The suramin taken up by T. brucei was recovered, in part, in the lysosomal fractions. It is suggested that deprivation of the parasite from cholesterol and phospholipids by an inhibition of the uptake of LDL, contributes to the mode of action of suramin, in addition to the many other effects that the drug may exert on the parasite. The toxic side-effects of suramin on the host are discussed in the light of its association with circulating lipoproteins.


Dictyocaulus viviparus: isolation and characterization of a recombinant antigen with potential for immunodiagnosis.
Crude adult worm antigen of Dictyocaulus viviparus was examined for specific antigens by SDS-PAGE and immunoblotting using sera from cattle experimentally infected with D. viviparus, vaccinated with a normal or a reduced dosage of the commercial lungworm vaccine, and helminth-free cattle. A D. viviparus-specific region M(r) 18,000 was identified and isolated. A lambda ZAP II cDNA expression library consisting of 4.4 x 10(5) recombinant clones (88% of the total number of clones) was constructed from D. viviparus adult worm mRNA. Rabbit antiserum to the M(r) 18,000 antigen was used to screen the cDNA library and eight positive clones were picked and allocated to the same antigenic family by sibling analysis. All clones were subcloned into the plasmid pGEX-2T, and the clone with highest expression yields was expressed as a glutathione S-transferase fusion protein (DvGST3-14) or, after cleavage with thrombin, as pure recombinant parasite protein (Dv3-14). The native parasite antigen encoded by the clone was identified. The immunodiagnostic potential of the recombinant proteins was assessed by immunoblotting.


HIV infection and AIDS.
Many of the clinical features of HIV/AIDS can be ascribed to the profound immune deficiency which develops in infected patients. The destruction of the immune system by the virus results in opportunistic infection, as well as an increased risk of autoimmune disease and malignancy. In addition, disease manifestations related to the virus itself may occur. For example, during the primary illness which occurs within weeks after first exposure to HIV, clinical symptoms occur in at least 50% of cases, typically as a mononucleosis syndrome. HIV-related complications are rarely encountered in patients with preserved immunity (i.e. CD4 T-cell counts greater than 500 cells/mm3). Recurrent mucocutaneous herpes simplex (HSV), herpes zoster (VZV), oral candidiasis and oral hairy leukoplakia occur with increasing frequency as the CD4 count drops below this level. Immune thrombocytopenia (ITP) occurs in association with HIV and often presents early in the clinical course. The risk of developing opportunistic infections and malignancies typical of AIDS increases progressively as CD4 counts fall below 200 cells/mm3. The clinical manifestations of infections associated with AIDS tend to fall into well-recognized patterns of presentation, including pneumonia, dysphagia/odynophagia, diarrhoea, neurological symptoms, fever, wasting, anaemia and visual loss. The commonest pathogens include Candida albicans, Pneumocystis carinii, Mycobacterium tuberculosis, Toxoplasma gondii, Cryptococcus neoformans, Mycobacterium avium intracellulare and cytomegalovirus. Malignant disease in patients with HIV infection also occurs in a characteristic pattern. Only two tumours are prevalent: Kaposi's sarcoma, a multifocal tumour of vascular endothelium which typically involves skin and mucosal surfaces; and non-Hodgkin's lymphoma, which is typically high grade in phenotype, often arising within the central nervous system. The principles of therapy include reduction of HIV replication by antiretroviral agents, prophylaxis against the common opportunistic infections and treatment followed by subsequent lifelong maintenance therapy for infections when they do occur.


Usefulness of heart-type fatty acid-binding protein in patients with severe sepsis.
The purpose of the study was to evaluate the value of heart-type fatty acid-binding protein (hFABP) as a novel clinical biomarker in patients with severe sepsis. Serum concentrations of hFABP and traditional cardiac biomarkers including cardiac troponin I, creatine kinase-MB, and B-type natriuretic peptides levels were measured within 6 hours after admission in 93 severe septic patients. The value of hFABP for the diagnosis of sepsis-related myocardial dysfunction (SRMD) and for the prediction of 28-day mortality was evaluated by receiver operating characteristics curve analysis. The prognostic value of elevated hFABP was subsequently confirmed by multivariate Cox proportional hazards analysis and Kaplan-Meier survival analysis. Heart-type fatty acid-binding protein was elevated (≥ 4.5 ng/mL) in 58 (62.4%) patients; patients with elevated hFABP appeared more likely to have SRMD (84.5% vs 31.4%, P < .001) and have higher prevalence of 28-day death (37.9% vs 8.6%, P = .002). Heart-type fatty acid-binding protein offered superior value over conventional biomarkers in both diagnosis of SRMD (area under the curve, 0.767; P < .001) and prediction of 28-day death (area under the curve, 0.805; P < .001). Serum hFABP is frequently elevated among patients with severe sepsis and appears to be associated with SRMD. Elevated hFABP independently predicts 28-day mortality in severe sepsis.


[Selective isolation and diversity of acidophilic filamentous actinomycetes from acidic soils].
To establish effective methods for selective isolation of acidophilic filamentous actinomycetes from acidic soils, and to investigate their genus and species diversity. Four pretreatments and 5 media supplemented with different inhibitors were used for isolation. The best combination of methods was determined according to the number and ratio of actinomycete colonies on plates, and was applied to isolate actinomycetes from 17 acidic soil samples collected in Jiangxi Province. The isolates were grouped based on cultural characteristics. The micromorphology and pH growth range of each group were studied to choose the representative isolates, which were subsequently subjected to 16S rRNA gene analysis to investigate their diversity. We found the best isolation approach involved Dispersion and Differential Centrifugation (DDC) pretreatment and GTV medium supplemented with cycloheximide, nalidixic acid nystatin (each at 50 mg/L). 369 isolates were obtained, and were assigned to 10 color groups. 6.6% of the isolates were strictly acidophilic actinomycetes, 72.4% were neutrotolerant acidophilic actinomycetes, and 21.0% were acidotolerant ones. The 52 representative isolates belong to 12 recognized genera: 30 of them fall within the genus Streptomyces, 6 belong to Micromonospora, 3 belong to Nocardia, 3 belong to Nonomuraea, 2 belong to Kribbella, 2 belong to Microbispora, and the other 6 strains belong to Actinomadura, Amycolatopsis, Dactylosporangium, Lentzea, Planotetraspora and Streptacidiphilus, respectivley. The Streptomyces isolates formed 12 evolutionary groups in the Streptomyces 16S rRNA gene tree. The selective isolation approach established here is robust for isolating various acidophilic filamentous actinomycetes from soil. Acidic soils in Jiangxi Province harbor abundant and diverse acidophilic actinomycetes.


Nucleus isthmi of the frog: structure and tecto-isthmic projection.
The morphology of neurons in the isthmic nucleus was studied with the Golgi technique. Most of the neurons have thick dendrites covered with lamelliform dendritic processes. The tecto-isthmic projection was investigated with the Fink--Heimer technique after partial tectal lesions. The anteromedial part of the tectum projects on the dorsal and anterior part, the caudomedial tectal region on the dorsal and posterior part of the nucleus. The posterolateral tectal area projects on the anterolateral part of the nucleus, and the axons originating in the anterolateral tectal quadrant terminate in its ventral and caudal part.


Prediction of intracranial pressure from noninvasive transocular venous and arterial hemodynamic measurements: a pilot study.
Continuous measurement of intracranial pressure (ICP) requires the invasive placement of epidural, parenchymal, or intraventricular devices. For critical single-point assessments, lumbar puncture may not always be practical. An accurate, reliable, portable and noninvasive method to estimate absolute ICP remains an elusive goal. The arteries that perfuse and the vein that drains the orbit are exposed to the ambient ICP while coursing through the cerebrospinal fluid or optic nerve. The venous outflow pressure (VOP) of the central retinal vein was measured using occlusion in six intensive care patients treated for acute hydrocephalus or brain hemorrhage and in whom transducers of intracranial pressure could provide standardized continuous output. A novel adaptation of the Balliart ophthalmodynamometer was developed for use. Simultaneously, the central retinal (CRA) and ophthalmic (OA) arterial flow velocities were recorded using color Doppler imaging technique. Repeat noninvasive measurements were performed at various ICPs, (n=22 independently collected observations). Linear regression and correlation testing were performed to evaluate these variables for ICP predictability. The VOP increased linearly with ICP (r=0.87). The arterial pulsatility indices for both OA and CRA decreased inversely with ICP (r=0.66). An empiric index combining both venous and arterial parameters (VOP/Gosling Pulsatility Index [GPI]) was significantly more correlated with absolute ICP than either parameter alone (r=0.95, p<0.005, ICP=0.29+0.74 [VOP/GPI(OA)]). The feasibility to estimate ICP from transocular sonographic and dynamometric data is suggested by these preliminary data. Retinal arterial properties are important in modeling the effect of ICP on the venous outflow pressure. Our pilot results serve as a basis on which to conduct a larger prospective and blinded study.


The effect of ranolazine on the vasodilator-induced myocardial perfusion abnormality.
We previously reported that ranolazine improved myocardial ischemia during exercise myocardial perfusion imaging (MPI). Since the mechanism of reversible perfusion defects is different in exercise than vasodilator MPI, and based on the mechanism of action of ranolazine, we hypothesized that vasodilator stress MPI may fail to show improvement in myocardial perfusion pattern. Patients (n = 18) with known coronary artery disease and with reversible perfusion defects on a clinically indicated vasodilator stress MPI were re-studied 4 weeks after ranolazine (500-1000 mg PO BID) was added to their conventional treatment in an open-label trial. Perfusion pattern was assessed using automated methods. The baseline left ventricular ejection fraction was 59% ± 14%. The total perfusion defect (measured by polar maps) was 22% ± 13% before and 21% ± 16% of LV myocardium after treatment (P = NS). The reversible defect size was 14% ± 10% before and 14% ± 11% of LV myocardium after treatment (P = NS). The automated-derived summed stress score was 12 ± 8 before and 12 ± 10 after treatment (P = NS) and the automated-derived summed difference score was 6 ± 5 before and 6 ± 5 after treatment (P = NS). Only 3 patients showed a decrease in reversible perfusion defect size with treatment. Vasodilator stress MPI failed to show improvement in perfusion pattern after ranolazine treatment in most patients with baseline reversible defects. This is consistent with the unique anti-ischemic mechanism of ranolazine, which acts on the late I Na channel.


Treatment of acyclovir-resistant herpetic ulceration with topical foscarnet and antiviral sensitivity analysis.
Herpes simplex virus (HSV) can produce persistent mucocutaneous disease in patients with the acquired immunodeficiency syndrome (AIDS). In this case report, we evaluate the efficacy, safety and viral resistance after topical foscarnet in severe genital ulceration due to acyclovir-resistant HSV-2. A 45-year-old African woman was known for an HIV infection with severe immunosuppression (CD4 <100/mm3). She had received a long-term prophylaxis with acyclovir (400 mg b.i.d.) for a recurrent genital herpes. Few weeks after stopping this prophylaxis, she developed large genital ulcerations progressing despite valacyclovir treatment (1,000 mg t.i.d.). Cultures were positive for HSV-2, resistance to acyclovir was shown by the plaque reduction assay and topical foscarnet was tried. Treatment consisted of a 20-min application of topical foscarnet 2.4% twice a day. Dramatic improvement was observed with rapid antalgia, and cicatrization of the genital ulcerations was observed after 50 days. HSV could not be detected on the mucosal surface. Initially, HSV-2 was resistant to acyclovir but sensitive to foscarnet. After 1 month of topical treatment, HSV-2 became sensitive to acyclovir and was still sensitive to foscarnet. Finally, after 6 weeks of treatment, no virus could be detected by culture. Topical foscarnet (2.4%) is a convenient treatment for chronic genital herpes. Resistance to acyclovir disappears few weeks after stopping this drug and sensitivity to foscarnet persists during the 50 days of treatment.


Dynamic charge acceptance and hydrogen evolution of a new MXene additive in advanced lead-acid batteries via a rapid screening three-electrode method.
A methodology to screen additives in lead-acid batteries is critical. We developed a three-electrode system that can rapidly check the dynamic charge acceptance (DCA) and hydrogen evolution of electrodes. The electrode with 2% MXene and 0.2% carbon black shows a better DCA, which indicates its great potential in the start-stop technology.


Effects of methylphenidate on sensitivity to reinforcement delay and to reinforcement amount in pigeons: Implications for impulsive choice.
Methylphenidate has been shown to decrease impulsive choice (increase choices of a larger more delayed reinforcer). The purpose of this study was to investigate 2 potential behavioral mechanisms of this effect: a drug-induced change in control by reinforcement delay (Experiment 1) and/or by reinforcement amount (Experiment 2). In Experiment 1, pigeons responded under a rapid-acquisition, concurrent-chains choice procedure involving delay to reinforcement; the option with the shorter delay varied unpredictably across sessions. The pigeons accurately tracked the shorter delay across sessions (i.e., a preference for the option with the shorter delay developed within each session). Methylphenidate selectively decreased sensitivity to reinforcement delay-it attenuated the acquisition of preference at doses that did not systematically affect bias or response rates. In Experiment 2, pigeons responded under a rapid-acquisition, concurrent-chains choice procedure involving reinforcement amount. The pigeons accurately tracked the option with the larger reinforcement amount across sessions. Methylphenidate selectively decreased sensitivity to reinforcement amount-it attenuated the acquisition of preference at doses that did not systematically affect bias or response rates. These data suggest that methylphenidate attenuates the degree to which the various reinforcement dimensions control choice, and that drug effects on impulsive choice depend upon the relative contributions of drug-induced changes in control of behavior by each relevant dimension. (PsycINFO Database Record


RNA interference highlights the role of CCaMK in dissemination of endosymbionts in the Aeschynomeneae legume Arachis.
In legume-rhizobia symbiosis, Ca2+/calmodulin-dependent protein kinase (CCaMK) is essential for rhizobial invasion through infection threads in the epidermis and nodule organogenesis in the cortex. Though CCaMK is actively transcribed in the infected zone of nodules, its role in the later stages of nodule development remain elusive because of the epidermal arrest of "loss-of-function" mutants. In Aeschynomeneae legumes such as Arachis hypogea, rhizobia directly access the cortex, where nodule organogenesis as well as endosymbiont dissemination take place by multiplication of infected cortical cells. We characterized CCaMK (GI:195542474) from A. hypogea and downregulated the kinase through RNA interference (RNAi) to understand its role during organogenesis of its characteristic aeschynomenoid nodules. In CCaMK downregulated plants, the inception of nodules was delayed by approximately 4 weeks and nodulation capacity was decreased (>90%). The infected zones of the RNA interference nodules were scattered with uninfected or binucleated cells as opposed to the homogeneous infection zone in empty-vector-transformed nodules. Symbiosomes in RNAi nodules were pleomorphic with diverse geometrical shapes or arrested during division in the final stages of their fission as opposed to uniform-sized, spherical symbiosomes in empty-vector-transformed nodules. Together, our results reveal CCaMK to be essential for development of functional aeschynomenoid nodules, with a critical role in rhizobial dissemination during nodule organogenesis.


Polymorphisms of transforming growth factor-β signaling pathway and Kawasaki disease in the Taiwanese population.
Kawasaki disease (KD) is a systemic vasculitis associated with cardiovascular symptom. A previous study in the European descent has indicated that genetic variants of the transforming growth factor-β (TGF-β) pathway are involved in the KD susceptibility and clinical status. This study was conducted to investigate if polymorphisms in TGF-β signaling pathway are associated with KD susceptibility, and the coronary artery lesion formation. A total of 950 subjects (381 KD patients and 569 controls) were investigated to identify 12 single-nucleotide polymorphisms in the TGF-β signaling pathway (rs2796817, rs10482751, rs2027567, rs12029576, rs11466480, rs4776338, rs12901071, rs7162912, rs1438386, rs6494633, rs12910698 and rs4776339) by using TaqMan Allelic Discrimination assay. Our results indicated that rs1438386 in the SMAD3 is significantly associated with the susceptibility of KD. Additionally, both haplotypes of TGFβ2 and SMAD3 were also associated with the risk of KD. This study showed that genetic polymorphisms in TGF-β signaling pathway are associated with KD susceptibility, but not coronary artery lesions formation, or intravenous immunoglobulin treatment response in the Taiwanese population.


Treatment for cramps in amyotrophic lateral sclerosis/motor neuron disease.
Cramps are painful, involuntary muscle contractions. They commonly affect people with amyotrophic lateral sclerosis/motor neuron disease (ALS/MND) at all stages of the disease. To date, the treatment of muscle cramps in ALS has been largely empirical without any evidence from randomised controlled trials. To systematically assess the effect of interventions on muscle cramps as a primary or secondary endpoint or adverse event in people with ALS/MND. We searched the Cochrane Neuromuscular Disease Group Specialized Register (14 February 2011), the Cochrane Central Register of Controlled Trials (Issue 1, 2011 in The Cochrane Library), MEDLINE (January 1966 to January 2011) and EMBASE (January 1980 to January 2011) and reference lists of articles searched using the terms motor neuron disease, motor neurone disease, motoneuron disease or amyotrophic lateral sclerosis. We contacted authors of trials for further information. We included all randomised and quasi-randomised trials of oral medications in people with ALS which assessed cramps as a primary or secondary outcome measure or as an adverse event. We also included trials using subcutaneous or intravenous medications or physical therapy. All authors applied the selection criteria and assessed study quality independently, and all authors performed independent data extraction. Twenty studies including 4789 participants were identified. Only one trial, of tetrahydrocannabinol (THC), assessed cramps as the primary endpoint. Thirteen studies assessed cramps as a secondary endpoint. The medications comprised vitamin E, baclofen, riluzole, L-threonine, xaliproden, indinavir, and memantine. Six studies assessed cramps as an adverse event. The medications comprised creatine, gabapentin, dextromethorphan, quinidine, and lithium. In all 20 studies no favourable effect for the treatment of cramps in ALS/MND could be demonstrated, but many studies were underpowered to draw a definite conclusion. A meta-analysis of two small studies showed a statistically nonsignificant result for the amino acid L-threonine for the treatment of cramps in ALS/MND. No study was identified using physical therapy as a therapeutic intervention for cramps. There is no evidence to support the use of any intervention for muscle cramps in ALS/MND. More and larger randomised controlled trials evaluating treatments for muscle cramps in ALS/MND are needed.


[Magnetic resonance imaging of atherosclerotic plaque].
Recent advances in terms of spatial and temporal resolution have enabled magnetic resonance imaging (MRI) to be used to display atherosclerotic plaque. MRI permits not only to detect atherosclerotic lesions but also enables determination of plaque volume and plaque characterization. It allows to display different plaque components such as lipid core, fibrous cap, calcium, and thrombus. To improve the spatial resolution, different invasive approaches based on intravascular coils have been evaluated. Novel contrast agent developments aim at the detection of inflammatory plaque activity in order to identify lesions with a high vascular risk (vulnerable plaque).


[A case of toxicity caused by drug interaction between capecitabine and phenytoin in patient with colorectal cancer].
We present a case of toxicity caused by a drug interaction between capecitabine and phenytoin (PHT). The drug combination elevated the plasma level of PHT in a patient on chemotherapy with capecitabine for colorectal cancer. Our patient was a 44-year-old woman diagnosed with epilepsy in her 20's, being treated with valproic acid (VPA) and PHT. Adjuvant chemotherapy with capecitabine began following surgery for colorectal cancer. Seven weeks later, she developed numbness, dizziness, dysarthria and difficulty walking, and was hospitalized for investigation. Her serum PHT level was elevated at 35. 1 μg/ mL. This case suggests that when capecitabine and PHT are coadministered, PHT levels should be monitored frequently, and that PHT dosage should be adjusted accordingly if it cannot be replaced by an alternative anticonvulsant.


The dark phase improves genetic discrimination for some high throughput mouse behavioral phenotyping.
Dark-phase testing has previously been shown by others to improve the outcome of some 'classical' behavior test situations. However, the importance of such ethological correctness and the effect of the light/dark cycle on high throughput behavioral testing situations such as 'mutant vs. wild type' and 'screening', are less or unknown, respectively. These testing situations differ from the 'classical' in that they are designed primarily to discriminate between genetically different mice rather than provide a detailed assessment of ability or psychosocial state. Here we test the hypotheses that dark-phase testing affects the outcome of high throughput behavioral tests and that dark-phase testing improves discrimination between genetically distinct mice (C57BL/6J, 129S1/SvImJ and B6129F1) using high throughput behavioral tests. Our results demonstrate that, although all successful tests showed some effect of phase, only the SHIRPA primary screen, open-field test and motor learning on the rotarod showed improved strain discrimination in the dark phase. Surprisingly, the social interaction test did not show a clear benefit to either phase, and interestingly, the tail-flick test discriminated strains better in the light phase. However, since the preponderance of our data shows that dark-phase testing improves, or does not affect, strain discrimination, we conclude that for these strains and tests, dark-phase testing provided superior outcomes. If discrimination is not achieved in the dark phase, then light phase-testing would be undertaken.


Factors determining renal transplant outcome at the University of Texas at Houston.
CsA-Pred therapy yields equivalently good patient survival for LRD and 2 degrees CAD versus 1 degree CAD transplants. There is a long-term graft survival advantage for LRD versus 1 degree CAD transplants (5 years; 83% vs 58%). 2 degrees CAD transplants have inferior graft survival when compared with 1 degree CAD grafts (one year; 78% vs 67%). Multiple donor factors adversely affecting graft outcome include increased warm and cold ischemia times, pulsatile perfusion, use of pressors or diuretics in the donor, donor age less than 10 years, donor blood transfusions, and kidneys shipped from other centers. Recipient factors adversely affecting graft outcome include retransplantation and CMV infection as well as noncompliance with therapy. HLA-matching and pretransplant blood transfusions have not contributed in a statistically significant way to graft outcome although they may affect the quality of graft function at this center. Immunosuppressive therapy with CsA-Pred must be tailored to the individual patient. Continuous IV CsA infusion in the preoperative period and slow steroid taper impact favorably on graft outcome. The complications of CsA therapy include neuroectodermal toxicity, hepatotoxicity, and most importantly, nephrotoxicity. Other problems unique to CsA-Pred therapy include hypertension, delayed graft thrombosis, and de novo hemolytic uremic syndrome. Hepatotoxicity may eventuate in biliary and pancreatic complications necessitating surgical therapy. The overall incidence of infection and neoplasm remains low with CsA-Pred therapy. The use of therapeutic trough CsA level monitoring, as well as pharmacokinetic and pharmacodynamic analyses may assist in clinical decision making regarding administered doses, dosing interval, and discrimination between rejection and nephrotoxicity.


CD137 activation abrogates granulocyte-macrophage colony-stimulating factor-mediated anti-apoptosis in neutrophils.
CD137 (ILA / 4-1BB) is a member of the TNF / NGF receptor family, and has previously been suggested to be involved in T cell activation and differentiation. Here, we demonstrate that blood neutrophils from control individuals and patients with cystic fibrosis express CD137 mRNA and surface protein. In contrast, lung neutrophils derived from patients with cystic fibrosis did not express detectable CD137 levels. Such CD137-deficient neutrophils could also be generated from normal neutrophils by TNF-alpha stimulation in vitro. TNF-alpha was found to be highly expressed in epithelial cells from cystic fibrosis but not normal lungs, suggesting that TNF-alpha might account for reduced neutrophil CD137 levels under inflammatory conditions in vivo. To investigate whether CD137 is involved in the regulation of apoptosis, neutrophils were activated with functional anti-CD137 antibody in the presence or absence of different neutrophil survival factors in vitro. Activation of CD137 abrogated GM-CSF-mediated anti-apoptosis in normal but not in CD137-deficient neutrophils. Moreover, G-CSF- and IFN-gamma-mediated neutrophil anti-apoptosis was not affected by anti-CD137 antibody treatment. In conclusion, these data suggest that CD137 activation may limit GM-CSF-mediated anti-apoptosis of neutrophils. The absence of this anti-inflammatory mechanism in inflammatory responses might be associated with massive neutrophil accumulation and consequent tissue damage.


The effects of serum albumin and phospholipid on sterol excretion in tissue culture cells.
Serum albumin was as effective as whole serum or alpha-globulins in facilitating sterol release from strain L mouse fibroblasts. Commercial bovine serum albumin preparations, however, had markedly different absolute effects in this regard. These differences were attributable to the variation in phospholipid content of these products. All but one of these albumins enhanced sterol release when supplemented with phospholipid. The exception was fatty acid-poor albumin which contained an adequate amount of phospholipid. Among the phospholipids examined, lecithin proved to be most effective, while phosphatidylethanolamine had little potentiating influence. As the unsaturation of the test lecithins increased, enhancement of sterol release decreased. The potentiating effect of the phospholipid was in turn dependent on the protein used, since the phenomenon was not observed with non-serum proteins like ovalbumin or with non-transport serum proteins such as gamma-globulins. The results of these studies raise the possibility that serum albumin together with phospholipid can play an important role in sterol release in tissue culture cells.


Breast Cancer 18F-ISO-1 Uptake as a Marker of Proliferation Status.
The sigma-2 receptor is a potential in vivo target for measuring proliferative status in cancer. Dehdashti et al. established the feasibility of using N-(4-(6,7-dimethoxy-3,-4-dihydroisoquinolin-2(1H)-yl)butyl)-2-(2-18F-fluoroethoxy)-5-methylbenzamide (18F-ISO-1) to image solid tumors in lymphoma, breast cancer, and head and neck cancer (1). Here we report results of the first dedicated clinical trial of 18F-ISO-1 in women with primary breast cancer. Our study objective was to determine whether 18F-ISO-1 PET could provide an in vivo measure of tumor proliferative status, and we hypothesized uptake would correlate with a tissue based assay of proliferation, namely Ki-67 expression. Methods: Twenty-eight women with 29 primary invasive breast cancers were prospectively enrolled in a clinical trial (NCT02284919) between 3/2015 and 1/2017. Each received an injection of 278-527 MBq 18F-ISO-1 followed by a 50-55 minute post-injection positron emission tomography-computed tomography (PET/CT) image of the breasts. In vivo uptake of 18F-ISO-1 was quantitated by maximum standardized uptake values (SUVmax) and distribution volume ratios (DVR) and was compared to ex vivo immunohistochemistry for Ki-67. Wilcoxon rank-sum test assessed uptake differences across Ki-67 thresholds and Spearman's correlation tested associations between uptake and Ki-67. Results: Tumor SUVmax (median 2.0 g/mL, range 1.3-3.3 g/mL), partial volume corrected (PVC) SUVmax, and SUV ratios were tested against Ki-67. Tumors stratified into the high Ki-67 (≥20%) group had SUVmax greater than the low Ki-67 (<20%) group (P = 0.02). SUVmax exhibited a positive correlation with Ki-67 across all breast cancer subtypes (ρ=0.46, P = 0.01, n = 29). PVC SUVmax was positively correlated with Ki-67 for invasive ductal carcinoma (ρ=0.51, P = 0.02, n = 21). Tumor-to-normal-tissue ratios and tumor DVR did not correlate with Ki-67 (P>0.05). Conclusion: 18F-ISO-1 uptake in breast cancer modestly correlates with an in vitro assay of proliferation.


Effects of coal dust contiguity on xerophthalmia development.
To study the occurrence of ocular surface diseases on coal miners working under wells and to investigate the relation between xerophthalmia occurrence and coal dust contiguity. To observe the influence of coal dust to ocular surface by using Schirmer to detect lacrimal gland secreting function, and breakup time of tear film (BUT) to evaluate the stability of lacrimal film and the quantity of lysozyme in lacrimal fluid. These values were lower for underground coal miners than other workers not contacting coal dust, and showed negative correlation to period of their services. Conditions in coal miners were worse than that of other workers. Coal dust exposure plays an important role in the development of dry eye.


Evaluation of the Roche LightCycler: a simple and rapid method for direct detection of factor V Leiden and prothrombin G20210A genotypes from blood samples without the need for DNA extraction.
The Roche LightCycler is a micro-volume thermocycler that combines extremely rapid polymerase chain reaction with fluorescence resonance energy transfer analysis of amplified products. We have evaluated the use of minimally processed blood samples for detection of two point mutations known to increase the risk of venous thromboembolism. Results from the LightCycler using the supernatant of diluted heated blood were compared with those gained by traditional methods based on polymerase chain reaction and restriction enzyme digestion. For factor V Leiden mutation, there was complete agreement between both methods in detection of wild-type (n = 82), heterozygous (n = 100) and homozygous (n = 18) genotypes. Similarly, the prothrombin G20210A mutation showed complete agreement for wild-type (n = 135), heterozygous (n = 63) and homozygous (n = 2) subjects.


Safety evaluation of lasalocid use in Chinese ring-necked pheasants (Phasianus colchicus).
Coccidiosis remains a significant threat to the welfare of game farm-reared pheasants in the United States. Although lasalocid has been demonstrated to be effective against pheasant specific coccidia, information regarding its safety in this species is lacking. The purpose of this study was to gather data on the safety of lasalocid when fed to Chinese ring-necked pheasants at one, two, and three times the recommended high dose of lasalocid used for prevention of coccidiosis in other poultry at three times the normal treatment period. Pheasant chicks (approximately 1 day-old; n = 160) were randomly blocked by sex into four treatment groups and given their respective diets continuously for 6 wk. No significant differences were observed in overall feed consumption, weight gain, feed conversion rates, clinical pathology measurements, or tissue gross and histopathologic evaluations between controls and treatment groups associated with lasalocid administration. Based on the results of this study it appears that lasalocid fed at the recommended rate of 125 ppm is safe in Chinese ring-necked pheasants.


A mass spectrometry-based probe of equilibrium intermediates in protein-folding reactions.
Described here is a mass spectrometry- and H/D exchange-based approach for the detection of equilibrium intermediate state(s) in protein-folding reactions. The approach utilizes the stability of unpurified proteins from rates of H/D exchange (SUPREX) technique to measure the m value (i.e., delta DeltaG/delta [denaturant] value) associated with the folding reaction of a protein. Such SUPREX m-value analyses can be made over a wide range of denaturant concentrations. Thus, the described approach is well-suited for the detection of high-energy intermediates that might be populated at low denaturant concentrations and hard to detect in conventional chemical denaturation experiments using spectroscopic probes. The approach is demonstrated on four known non-two-state folding proteins, including alpha-lactalbumin, cytochrome c, intestinal fatty acid binding protein (IFABP), and myoglobin. The non-two-state folding behavior of each model protein system was detected by the described method. The cytochrome c, myoglobin, and IFABP systems each had high-energy intermediate states that were undetected in conventional optical spectroscopy-based studies and previously required other more specialized biophysical approaches (e.g., nuclear magnetic resonance spectroscopy-based methods and protease protection assays) for their detection. The SUPREX-based approach outlined here offers an attractive alternative to these other approaches, because it has the advantage of speed and the ability to analyze both purified and unpurified protein samples in either concentrated or dilute solution.


Detection of free antielastin antibodies among diabetic children.
Antibodies to elastin breakdown products are found in the serum of all human subjects and correlate with their respective serum peptide levels. The presence of these antielastin antibodies (AEAbs) and the corresponding antigens in circulation leads to the formation of circulating immune complexes (CICs). The aim of this study was to determine if the serum levels of free AEAbs (not bound in CICs) correlate with the development of vascular complications in diabetic children. To this end, we used a method for detecting immune complexes (complement inhibition factor [CIF]-enzyme-linked immunosorbent assay [ELISA]) in combination with an ELISA for detection of AEAbs. The levels of free immunoglobulin G (IgG) AEAbs were studied in the sera of 54 diabetic children (mean age 12.3+/-4 years; diabetes duration 5.2+/-3.7 years). Thirty-two of the children had vascular complications (group 1), and 22 were without vascular complications (group 2). Twenty healthy children (mean age 13.6+/-4.2 years) were used as controls. The diabetics showed statistically significant higher levels of free AEAbs (0.490 E492+/-0.244 E492 vs 0.307 E492+/-0.081 E492; p = .02) compared with the control group. In group 1, free AEAbs showed statistically significant higher levels than controls (0.523+/-0.269 vs 0.307+/-0.081; p = .016). Eighteen of 54 (33%) patients were positive for free AEAbs (13 of 32 [41%] in group 1 and 5 of 22 [22%] in group 2). Free AEAb levels in all diabetics showed a correlation with systolic blood pressure (r = .44; p = .01), diastolic blood pressure (r = .46; p = .009), total cholesterol (r = .33; p = .05), triglycerides (r= .38; p = .03), high-density lipoprotein (r= -.46; p = .009), serum fructose (r= .43; p = .001), and microalbuminuria (r= .41; p = .002). Patients who had vascular pathology showed a correlation of free AEAbs with microalbuminuria (r= .434; p= .026), serum fructose (r= .63; p = .0004), hemoglobin A1c (r= .392; p = .043), and triglycerides (r= .456; p = .025). These findings suggest that elevated levels of free IgG AEAbs are associated with the development of diabetic vascular complications in children.


Gender differences in serum angiotensin-converting enzyme activity and blood pressure in children: an observational study.
Angiotensin-converting enzyme (ACE) is a key enzyme of the renin-angiotensin system that plays an important role in regulating blood pressure. ACE enzyme activity and its relationships with blood pressure (BP) during childhood and adolescence have not yet been clearly established. To determine serum ACE (S-ACE) levels and BP changes in school children between 8 and 18 years of age and how S-ACE and BP in males and females might differ, as well as to determine S-ACE and BP relationships. Blood pressure, height, weight, body mass index (BMI), and S-ACE were measured in 501 children. Mean S-ACE values were higher in boys (143.7+/-57.1) than in girls (130.2 +/- 54.9) (p = 0.004). S-ACE values decreased in girls and increased in boys with age, and values for girls were lower than for age-matched boys after onset of puberty. Age was a strong determinant of BP levels in both genders. We found a relationship between ACE and systolic blood pressure (SBP) and diastolic blood pressure (DBP) in girls (SBP r= -0.20 p<0.001 DBP r=0.12 p<0.03). BMI had greater correlation with SBP and DBP in girls (r=0.37, and 0.31, respectively; p < 0.001) than in boys (r=0.26, and 0.25 respectively; p<0.001). These results indicate that gender differences in serum ACE activity exist in the children from this study. This activity was lower and decreased with age in girls, while BP increased. Because sexual dimorphism in BP emerges in puberty, our findings suggest that gonadal hormones might affect S-ACE activity and BP. These results may have important therapeutic implications.


The ING1a model of rapid cell senescence.
Replicative capacity of normal human cells decreases as telomeric sequence is lost at each division. It is believed that when a subset of chromosomes reach a critically short length, an ATM-initiated and p53-mediated transcriptional response inhibits cell growth, promoting cell senescence. In addition to loss of telomeric sequence, senescence can be induced by other stresses including ionizing radiation, oxidative damage, chemical crosslinkers like the chemotherapeutic agent cisplatin, as well as overactivation of oncogenes and tumor suppressors. Our group found that the expression of an isoform of the INhibitor of Growth 1 gene called ING1a increases approximately 10-fold as fibroblasts approach senescence and that forced expression rapidly induces a senescent phenotype in primary diploid fibroblasts, epithelial and endothelial cells that resembles replicative senescence by most physical and biochemical measures. ING1a induces these changes through strongly inhibiting endocytosis to block mitogen signaling by inducing the expression of intersectin 2, a key scaffolding protein of the endosomal pathway. This, in turn increases the expression of Rb and of p57Kip2 and p16INK4a that serve to maintain Rb is an active, growth inhibitory state. The ING1a model is currently being used to better understand the mechanism(s) responsible for activating Rb to enforce the senescent state.


Prevention of hypomagnesemia and hypocalcemia in open-heart surgery.
We have studied the behavior of calcium and magnesium in the serum in dogs subjects to cardiopulmonary bypass with total hemodilution (Ringer's lactate). 5 mEq/l of calcium and magnesium was added to the filling solution. Serum levels of Ca++ and Mg++ were determined, by atomic absorption spectrophotometry, in the basal state, every thirty minutes during the perfusion, and one hour after stopping the bypass. We observed that the calcium and magnesium levels were higher than be baseline throughout the perfusion, and even 60 minutes after stopping the bypass they were still noticeably higher. Since this type of perfusion yields normal final calcium and magnesium levels, the advantages of this method become obvious, especially considering the adverse phenomena that are caused by defiencies of these cations.


Neonatal nonimmune hemolytic anemia.
As in adults and older children, anemia in newborn infants can be the result of erythropoietic failure, hemorrhage, or hemolysis. When hemolysis is the prime consideration, it can be challenging for physicians caring for neonates to choose from the wide variety of available diagnostic tests. This review describes the authors' opinions regarding rational, consistent, and cost-effective means of making an exact diagnosis of a neonatal hemolytic condition. Two recent advances in the diagnosis of neonatal nonimmune hemolytic disorders are highlighted in this review: introduction of flow cytometry-based Eosin-5-maleimide (EMA) uptake as a screening test to identify RBC membrane defects and next-generation sequencing (NGS)-based panels to uncover exact mutations causing hemolysis. The availability of newer tools such as EMA and NGS to diagnose specific hemolytic conditions, which might otherwise remain unknown, enables neonatal practitioners not only to identify the exact cause of hemolysis but also to discover novel mutations that can be implicated in the cause of neonatal hemolytic processes.


Persistent eIF2alpha(P) is colocalized with cytoplasmic cytochrome c in vulnerable hippocampal neurons after 4 hours of reperfusion following 10-minute complete brain ischemia.
Upon brain reperfusion following ischemia, there is widespread inhibition of neuronal protein synthesis that is due to phosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha), which persists in selectively vulnerable neurons (SVNs) destined to die. Other investigators have shown that expression of mutant eIF2alpha (S51D) mimicking phosphorylated eIF2alpha induces apoptosis, and expression of non-phosphorylatable eIF2alpha (S51A) blocks induction of apoptosis. An early event in initiating apoptosis is the release of cytochrome c from mitochondria, and cytochrome c release corresponds to the selective vulnerability of hippocampal CA1 neurons in rats after transient global cerebral ischemia. At present the signaling pathways leading to this are not well defined. We hypothesized that persistent eIF2alpha(P) reflects injury mechanisms that are causally upstream of release of cytochrome c and induction of apoptosis. At 4 h of reperfusion following 10-min cardiac arrest, vulnerable neurons in the striatum, hippocampal hilus and CA1 showed colocalized intense immunostaining for both persistent eIF2alpha(P) and cytoplasmic cytochrome c, while resistant neurons in the dentate gyrus and elsewhere did not immunostain for either. A lower intensity of persistent eIF2alpha(P) immunostaining was present in cortical layer V pyramidal neurons without cytoplasmic cytochrome c, possibly reflecting the lesser vulnerability of this area to ischemia. We did not observe cytoplasmic cytochrome c in any neurons that did not also display persistent eIF2alpha(P) immunostaining. Because phosphorylation of eIF2alpha during early brain reperfusion is carried out by PERK, these findings suggest that there is prolonged activation of the unfolded protein response in the reperfused brain.


Modified Friedman technique: a new proposed method of measuring glenoid version in the setting of glenohumeral dysplasia.
Glenoid version angles are measured to objectively follow changes related to glenohumeral dysplasia in the setting of brachial plexus birth palsy. Measuring glenoid version on cross-sectional imaging was initially described by Friedman et al. in 1992. Recent literature for non-dysplastic shoulders advocates time-consuming reconstructions and reformations for an accurate assessment of glenoid version. To compare Friedman's original method for measuring glenoid version to a novel technique we developed ("modified Friedman") with the reference standard of true axial reformations. With institutional review board approval, we retrospectively examined 30 normal and dysplastic shoulders obtained from magnetic resonance imaging examinations of 30 patients with an established diagnosis of brachial plexus birth palsy between January 2012 and September 2017. Four pediatric radiologists performed glenoid version measurements using Friedman's method, the modified Friedman method and a previously described true axial reformation method. The modified Friedman technique better accounts for scapular positioning by selecting a reference point related to the acromion-scapular body interface. Inter-rater reliability and inter-method agreement were assessed using intraclass correlation, paired t-tests and mixed linear model analysis. Equivalence tests between methods were performed per reader. Glenoid version measurements were significantly different when comparing Friedman's method to true axial reformations in normal (-10.8±5.7° [mean±standard deviation] vs. -8.8±5.3°; P≤0.001) and dysplastic shoulders (-34.6±17.7° vs. -28.1±17.5°; P≤0.001). Glenoid version measurements were not significantly different when comparing the modified Friedman's method to true axial reformations in normal (-6.3±5.8° vs. -8.8±5.3°; P=0.06) and dysplastic shoulders (-29.0±18.3° vs. -28.1±17.5°; P=0.06). Friedman's method was not equivalent to true axial reformations for measurements in dysplastic shoulders for all readers (P=0.68, 0.81, 0.86, 0.99); the modified Friedman method was equivalent to of true axial reformations for measurements in dysplastic shoulders for 3 of 4 readers (P≤0.001, P≤0.001, P≤0.001, P=0.10). In glenohumeral dysplasia, the modified Friedman method and post-processed true axial reformations provide statistically similar and reproducible values. We propose that our modified Friedman technique can be performed in lieu of post-processed true axial reformations to generate glenoid version measurements.


Chirality transfer from organic ligands to silver nanostructures via chiral polarisation of the electric field.
Water-soluble ligand protected optically active silver nanostructures were synthesised in a one-step reduction and capping process mediated by thiol-containing biomolecules. The synthesis was performed successfully with d- and l-cysteine and l-glutathione. The chiroptical properties of the obtained nanostructures were investigated by circular dichroism spectroscopy in the ultraviolet and visible wavelength range. They exhibit a g-value of up to 0.7%, which is about one order of magnitude larger compared to particles prepared by citrate reduction followed by a ligand exchange reaction. The structure and composition of the prepared materials were characterised by transmission electron microscopy, energy-dispersive X-ray and X-ray photoelectron spectroscopy. Although these structures do not have a chiral geometry, they show mirror image g-values when capped with d- and l-cysteine. This indicates that the underlying chirality transfer mechanism is based on an electric field polarisation process.


Structural effects of amino acid variations between B and CRF02-AG HIV-1 integrases.
HIV-1 integrase is one of the three essential enzyme required for viral replication and has a great potential as a novel target for anti-HIV drugs. The sequence variability of the entire integrase (IN) was examined in HIV-1 subtype B and CRF02-AG antiretroviral naïve infected patients for the presence of naturally occurring polymorphisms IN gene sequences and protein structures from both subtypes were compared. The phylogenetic analysis showed a total concordance between the 3 pol gene sequences for patients identified as subtype B whereas 3% of patients identified as CRF02-AG showed a mixture of subtypes. The analysis of IN aa sequences showed that 13 positions (K/R14, V/I31, L/I101, T/V112, T/A124, T/A125, G/N134, I/V135, K/T136, V/I201, T/S206, L/I234, and S/G283) differed between subtypes B and CRF02-AG. As observed in the 3D model of the preintegration complex, these differences may impact the functional property of IN. The fact that most variations were grouped suggests that some of them are linked together through compensatory mechanisms. This comparison allowed us to identify several variations of amino acids in HIV-1 IN subtype CRF02-AG that could have a putative impact on anti-integrase sensitivity. In particular, the region formed by Thr125, Thr124, Val31 contains at least one residue, T125, which variation has been involved in eliciting resistance to the naphtyridine carboxamide L870,810 IN inhibitor. In conclusion, virological response to anti-integrase should be studied carefully, according to the subtype, in clinical trials.


Rpi-blb2-mediated late blight resistance in Nicotiana benthamiana requires SGT1 and salicylic acid-mediated signaling but not RAR1 or HSP90.
The Rpi-blb2 recognizes the presence of the Phytophthora infestans AVRblb2 and initiates effector-triggered immunity (ETI). We performed gain-of-function and loss-of-function studies in Nicotiana benthamiana to elucidate Rpi-blb2-mediated resistance to P. infestans. Rpi-blb2 triggered a hypersensitive response through SGT1-mediated, but not RAR-mediated or HSP90-mediated, pathways. NbSGT1 was also required for basal and ETI-mediated by Rpi-blb2 in N. benthamiana. Moreover, salicylic acid (SA) affected basal defense and Rpi-blb2-mediated resistance against P. infestans. The increased susceptibility of Rpi-blb2-transgenic plants in the NahG-background correlated with reduced levels of SA. These findings provide evidence for the roles of SGT1- and SA-signaling in Rpi-blb2-mediated resistance against P. infestans.


Intracoronary optical coherence tomography for the assessment of in-stent restenosis.
Optical coherence tomography (OCT) is a novel imaging modality in interventional cardiology. With its clear image quality, it has attracted considerable attention as a tool to accurately evaluate atherosclerosis and coronary stents in vivo. It has a 10-fold higher resolution compared to intravascular ultrasound and utilises near-infrared light that requires temporary blood clearance during image acquisition. We present a case of in-stent restenosis in a patient five years after percutaneous intervention. Optical coherence tomography clearly showed the mechanism behind the stent failure and helped appropriately plan the subsequent repeat intervention. The technology heralds an exciting advance in the invasive assessment of the coronary arteries that will improve our understanding of heart disease into the future.


Dose impact of rectal gas on prostatic IMRT and VMAT.
In this study, we compared the dose impact of the heterogeneity caused by rectal gas using two methods of treatment planning for intensity-modulated radiotherapy (IMRT) and volumetric-modulated arc therapy (VMAT). In addition to the structure set used for the standard treatment plan, we created a structure set for evaluation for each patient. These sets were transferred to the same isocenter as the respective treatment plans for IMRT and VMAT that were to become the standard. Values were then recalculated. During the standard prostatic IMRT and VMAT treatment planning, all study participants met dose restrictions in place at our hospital. Dose restrictions were fulfilled in treatment plans for evaluation, excluding those with a clinical target volume (CTV) of V(100%) and planning target volume (PTV) of D95 when the rectum was excluded. However, in treatment plans for evaluation, IMRT was shown to have a higher concordance rate with standard treatment plans than VMAT. If rectal gas is present during either IMRT or VMAT, a dose decrease will occur in relation to CTV and PTV, suggesting that a plan does not eliminate adverse effects on organs at risk.


Early warning and NHS Direct: a role in community surveillance?
NHS Direct is a nurse-led telephone help line that covers the whole of England and Wales. NHS Direct derived data are being used for community surveillance, the purpose of which is to detect a local or national increase in symptoms reported by callers. The system has the potential to identify an increase in symptoms reported by callers about people in the prodromal stages of illness caused by the deliberate release of a biological or chemical agent. There are no other community surveillance projects existing on a national scale that utilize electronic daily data. We describe the surveillance system and calls to NHS Direct between December 2001 and July 2002. Confidence limits have been constructed for 10 key algorithms at each site and control charts devised for five of these algorithms at sites covering the key urban areas. Daily reporting has been achieved from NHS Direct sites in England and Wales. High levels of activity in specific algorithms at both national and regional levels have been detected. A sustained national increase in calls about fever occurred in January 2002. Although the project is still at an early stage, daily analysis of NHS Direct data has the potential to detect symptoms in the community that could be related to deliberate releases of chemical or biological agents or to outbreaks of disease. For this surveillance to act as an 'early warning' of illness resulting from a microbiological or chemical cause, the NHS Direct surveillance needs to be fully integrated into an appropriate public health response (which may require diagnostic samples to be taken from callers).


A simple method for isolation and characterization of mouse brain microvascular endothelial cells.
Brain endothelial cells, a site of the blood-brain barrier in vivo, regulate a number of physiological and pathophysiological processes in the brain including transport of nutrients, export of critical toxins, transmigration of circulating leukocytes and formation of new blood vessels. In this report, we describe a simple and reproducible method to isolate pure (>99%), functionally active endothelial cells from small quantities of adult mouse brain tissue. In vitro, these cells express typical phenotypic markers of differentiated brain endothelium such as von Willebrand factor, multiple drug resistant protein and glucose transporter-1, demonstrate uptake of acetylated low-density lipoprotein, and possess morphological and ultrastructural characteristics of microvascular endothelium. They form tight junctions and capillary-like tubes when stimulated by growth factors in an in vitro angiogenesis assay. In response to tumor necrosis factor-alpha, isolated mouse brain endothelial cells (MBEC) express vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). The protocol described here provides an effective and reliable method to isolate pure cerebral endothelium from adult mouse brain that should offer a useful tool for studying the role of altered vascular biology in mice with genetically manipulated brain disorders.


Solvatochromic behavior of the electronic absorption spectra of some azo derivatives of amino pyridines.
The electronic absorption spectra of a series of the entitled eight compounds containing groups with variable electronic characters were recorded. The solvents were selected to cover a wide range of parameters (refractive index, dielectric constant and hydrogen bonding capacity). The electronic transitions are assigned and the solvent induced spectral shifts have been analyzed in relation to the different solute-solvent interaction mechanisms using computational chemistry. The regression analysis is applied for correlation parameters. The phenomenon of tautomerism is explained. The electronic character of the substituent as well as the chemistry of the solvent are the major factors for the solvatochromic behavior.


Changes in the gastrointestinal tract, cardiovascular function and some drug metabolizing processes in rats and guinea-pigs intoxicated with aflatoxin B1.
The effects of aflatoxin B1 (AFB1) on the reactivity of the peripheral vessels, passage through the gastrointestinal system, production of hydrochloric acid and some liver functions were analyzed. The results shown the decreased glucuronidizing ability of the liver (58% of the initial value) accompanied by an increased amount of the unconjugated substrate while glycine conjugation was not changed in animals on the 3rd day after the treatment with AFB1. The uptake of 99mTc-SnO2 colloid was 1.14 times lower in the liver and 1.94 times higher in the spleen in AFB1 treated animals. A lowered activity of the glucuronizing system and a possibly indirectly lowered uptake of tin colloid in the liver indicate the toxic effect of AFB1 in hepatocytes. The other parameters studied were not changed or were changed only slightly.


Magnetic anal sphincter augmentation in patients with severe fecal incontinence.
Fecal incontinence is a common, distressing condition with limited surgical options. This study examines the results of magnetic sphincter augmentation in patients with severe fecal incontinence. This was a single-center, prospective, nonrandomized investigation. This study was conducted in a private colorectal practice. The cohort included all of the patients implanted with magnetic sphincter augmentation between January 2012 and October 2013. Magnetic sphincter augmentation was studied. Adverse events, symptom severity, quality of life, bowel diary, and manometry data were collected. Eighteen patients (15 women), with mean age of 69 years (range, 31-91 years), were implanted with magnetic sphincter augmentation. Follow-up ranged from 353 to 738 days. Previous treatment consisted of peripheral nerve evaluation test in 10 patients (56%), 2 patients (11%) with previous permanent sacral nerve stimulation, and 1 patient (6%) with previous implantation of an artificial bowel sphincter. Implantation was successful in 17 (94%) of 18 patients. Five patients (29%) had postoperative pain, and 5 patients (29%) had temporary swelling and erythema in both gluteal regions after the implantation. No devices were explanted during the follow-up. Cleveland Clinic Incontinence Score decreased from a mean of 17.5 (range, 14.0-20.0) to 7.3 (range, 0-12.0), and Fecal Incontinence Quality of Life scores improved in all of the domains. Bowel diary results showed that 76% of the patients with implants experienced a ≥50% reduction in the number of fecal incontinence episodes per week. Manometry at 6 months after implantation showed increased mean resting and squeeze pressures. This study does not allow for comparison between surgical treatments and involves a limited number of patients. Magnetic sphincter augmentation shows consistent results for the treatment of severe fecal incontinence in this patient group. The surgical procedure is straightforward as compared with other implantable devices. The safety profile is acceptable. Magnetic sphincter augmentation is a promising new treatment with the potential to become a first-line surgical therapy for patients with severe fecal incontinence.


Congenital visual agnosia and prosopagnosia in a child: a case report.
We present an eight years old child, L.G. with congenital agnosia and prosopagnosia. This is a special case of perceptual deficits in a child which are discrete and exist in the context of a very high verbal intelligence. L.G. was administered an extensive battery of tests of cognitive functioning. He has intact basic visual skills, although his visual analysis is sometimes slow. L.G. can read, write and do math at age level or above. Four normal eight years old boys were used as controls on a selection of the perceptual tests, administered to L.G., which did not have normative data. L.G.'s object recognition skills bear the hallmarks of adult apperceptive agnosia. His visual memory and imagery are normal. Tests of face processing skills reveal, unlike adult prosopagnosics, severe deficits in addition to the familiar face recognition problem. L.G.'s agnosia and prosopagnosia are compared to the relevant literature.


Discrete-state representation of ion permeation coupled to fast gating in a model of ClC chloride channels: comparison to multi-ion continuous space Brownian dynamics simulations.
A discrete-state model of chloride ion motion in a ClC chloride channel is constructed, following a previously developed multi-ion continuous space model of the same system (Cheng, M. H.; Mamonov, A. B.; Dukes, J. W.; Coalson, R. D. J. Phys. Chem. B 2007, 111, 5956) that included a simplistic representation of the fast gate in this channel. The reducibility of the many-body continuous space to the eight discrete-state model considered in the present work is examined in detail by performing three-dimensional Brownian dynamics simulations of each allowed state-to-state transition in order to extract the appropriate rate constant for this process, and then inserting the pairwise rate constants thereby obtained into an appropriate set of kinetic master equations. Experimental properties of interest, including the rate of Cl(-) ion permeation through the open channel and the average rate of closing of the fast gate as a function of bulk Cl(-) ion concentrations in the intracellular and extracellular electrolyte reservoirs are computed. Good agreement is found between the results obtained via the eight discrete-state model versus the multi-ion continuous space model, thereby encouraging continued development of the discrete-state model to include more complex behaviors observed experimentally in these channels.


Sleep Origins: Restful Jellyfish Are Sleeping Jellyfish.
What is the 'simplest' animal that sleeps? When did sleep first evolve? Do all animals sleep? Tantalizing hints to answers come from new research showing that jellyfish, one of the earliest evolving groups of animals, have a sleep-like restful state.


Radiation and the regulatory landscape of neo2-Darwinism.
Several recently revealed features of eukaryotic genomes were not predicted by earlier evolutionary paradigms, including the relatively small number of genes, the very large amounts of non-functional code and its quarantine in heterochromatin, the remarkable conservation of many functionally important genes across relatively enormous phylogenetic distances, and the prevalence of extra-genomic information associated with chromatin structure and histone proteins. All of these emphasize a paramount role for regulatory evolution, which is further reinforced by recent perspectives highlighting even higher-order regulation governing epigenetics and development (EVO-DEVO). Modern neo2-Darwinism, with its emphasis on regulatory mechanisms and regulatory evolution provides new vision for understanding radiation biology, particularly because free radicals and redox states are central to many regulatory mechanisms and free radicals generated by radiation mimic and amplify endogenous signalling. This paper explores some of these aspects and their implications for low-dose radiation biology.


Pathophysiology of Degenerative Mitral Regurgitation: New 3-Dimensional Imaging Insights.
Despite its high prevalence, little is known about mechanisms of mitral regurgitation in degenerative mitral valve disease apart from the leaflet prolapse itself. Mitral valve is a complex structure, including mitral annulus, mitral leaflets, papillary muscles, chords, and left ventricular walls. All these structures are involved in physiological and pathological functioning of this valvuloventricular complex but up to now were difficult to analyze because of inherent limitations of 2-dimensional imaging. The advent of 3-dimensional echocardiography, computed tomography, and cardiac magnetic resonance imaging overcoming these limitations provides new insights into mechanistic analysis of degenerative mitral regurgitation. This review will detail the contribution of quantitative and qualitative dynamic analysis of mitral annulus and mitral leaflets by new imaging methods in the understanding of degenerative mitral regurgitation pathophysiology.


Risk factors for recurrent stenosis following successful coronary angioplasty.
The major limitation of coronary angioplasty is recurrent stenosis. Patient, clinical, and procedural factors at the time of angioplasty were correlated with the presence or absence of angiographically documented recurrent stenosis or continued patency. Patients with single-vessel, multilesion disease had a lower incidence of recurrence than those with single-vessel, single-lesion disease (22.2% v 37.3%). Patients with multivessel disease had the highest rate of recurrent stenosis (45%). Sex and age did not predict recurrence. Patients with more severe symptoms, patients without prior myocardial infarction (MI), and insulin-dependent diabetic patients were more likely to have recurrent stenosis. Of the lesion variables, a severe stenosis or a low gradient before angioplasty, the absence of an intimal tear after angioplasty, and left anterior descending artery lesions correlated with a higher recurrence rate. Inflation times greater than or equal to 30 seconds and a greater total number of inflations correlated with a higher recurrence rate. Multivariate analysis showed the following variables to be important predictors of recurrence: Canadian Heart Class, history of myocardial infarction, gradient before angioplasty, artery dilated, number of inflations, severity of stenosis before angioplasty, and insulin-dependent diabetes mellitus.


Effect of chronic angiotensin II inhibition on the nitric oxide synthase in the normal rat during aging.
To assess the effect on the cardiovascular system, of enalapril (E) or losartan (L) given since weaning during 6 or 18 months to normal rats. Animals were divided in three groups: control (C), E-treated and L-treated; treated rats received 10 mg/ kg per day of drug. Systolic blood pressure (SBP), body weight, water and food intake (WI, FI), cardiac, left ventricular and aortic weight as well as the length of the tail were recorded. NADPH-diaphorase activity was determined as a marker of nitric oxide synthase (NOS) activity in aorta, arterioles of small intestine, heart and kidney of normal rats. NOS activity was measured as optical density (OD) in the stained tissue. Nitrate + nitrite urinary excretion was measured in 24 h urine. Only significant differences (P < 0.05) are reported. SBP, absolute cardiac, left ventricular and aortic weight increased with age. Both treatments delayed these increments. At 6 and 18 months, NOS activity was higher in aortic endothelium (Em) of L- and E-treated animals. Losartan treatment during 6 months also increased NOS activity in aortic smooth muscle (SM). Aortic Em NOS activity fell in the 18 months-treated and untreated animals. E increased NOS activity in the SM of intestinal arterioles at 6 months but reduced it at 18 months. The fact that both E and L delayed cardiac hypertrophy/hyperplasia and aortic growth and raised aortic endothelium NOS activity indicates a protective effect on cardiovascular damage due to aging, exerted through inhibition of angiotensin II.


Fine-needle aspiration and core needle biopsy: An update on 2 common minimally invasive tissue sampling modalities.
Fine-needle aspiration (FNA) and core needle biopsy (CNB) represent 2 of the most common minimally invasive tissue sampling modalities. Although similar in many ways, there are significant differences in the collection, processing, interpretation, and suitability for ancillary testing that exist between FNA and CNB. This review provides a brief overview of the strengths and weaknesses of FNA compared with CNB, as well as an update regarding the landscape of recently published studies that investigate the organ-specific comparative performance metrics of FNA and CNB. A current understanding of the benefits and limitations of FNA and CNB will help the cytopathologist and the clinician alike to select the right procedure for the right patient at the right time. Cancer Cytopathol 2016;124:862-870. © 2016 American Cancer Society.


When touch is not the best approach.
Health care professionals have long believed in the value of nonverbal components of communication and the highly significant roles such factors as distance, space, and touch can play in therapeutic relationships. The value of touch is not appreciated by all health care professionals or considered appropriate or desirable by some patients. While touch has been described as the most important of all the senses, the astute professional must be cognizant of the times when touch should not be used.


Cancer and adenomatous polyp distribution in the colorectum.
To examine the possible role of adenomatous polyps in colorectal cancer development, this study focused on the relationship of the distribution between adenomatous polyps and cancer in the colorectum. The distribution of 753 adenomatous polyps was compared with that of 35 colorectal cancer lesions in 510 male patients ranging from 45 to 55 yr of age who underwent a total colonoscopy. The incidence of cancer significantly increased with a distal shift in the colorectal sites in comparison with that of adenomatous polyps (p < 0.02). The different distribution between adenomatous polyps and cancer thus suggested that adenomatous polyps at various colorectal sites appear to have a different malignant potential for cancer development.


VASCULAR ABNORMALITIES IN DIABETIC RETINOPATHY ASSESSED WITH SWEPT-SOURCE OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY WIDEFIELD IMAGING.
To detect vascular abnormalities in diabetic retinopathy using swept-source optical coherence tomography angiography (SS-OCTA) widefield images, and to compare the findings with color fundus photographs (CFPs) using Early Treatment Diabetic Retinopathy Study severity grading. 3 mm × 3 mm and 12 mm × 12 mm scans were acquired to cover 70° to 80° of the posterior pole using a 100-kHz SS-OCTA instrument. Two masked graders assessed the presence of vascular abnormalities on SS-OCTA and the Early Treatment Diabetic Retinopathy Study level on CFP. The grading results were then compared. A total of 120 diabetic eyes (60 patients) were imaged with the SS-OCTA instrument. Cohort 1 (91 eyes; SS-OCTA grading only) showed microaneurysms in 91% (n = 83), intraretinal microvascular abnormalities in 79% (n = 72), and neovascularization in 21% (n = 19) of cases. Cohort 2 (52 eyes; CFP grading compared with SS-OCTA) showed microaneurysms on CFP in 90% (n = 47) and on SS-OCTA in 96% (n = 50) of cases. Agreement in intraretinal microvascular abnormality detection was fair (k = 0.2). Swept-source optical coherence tomography angiography detected 50% of intraretinal microvascular abnormality cases (n = 26), which were missed on CFP. Agreement in detecting neovascularization was moderate (k = 0.5). Agreement in detection of diabetic retinopathy features on CFP and SS-OCTA varies depending on the vascular changes examined. Swept-source optical coherence tomography angiography shows a higher detection rate of intraretinal microvascular abnormalities (P = 0.039), compared with Early Treatment Diabetic Retinopathy Study grading.


Dipyridamole, an underestimated vascular protective drug.
Dipyridamole, developed almost half a century ago, acts by inhibiting nucleoside transport, which increases adenosine levels leading to inhibition of platelet aggregation and vasodilatation mainly in the coronary tree. It is a vaso-protective drug with proven efficacy in the prevention of strokes. Adenosine receptor 2 inhibitory purines, ubiquitously available in food and drink, inhibit the vasomotor effects of dipyridamole but not its action on platelet aggregation. This and the slow build-up of blood levels of dipyridamole after oral application may explain why incidents of drug-induced angina ("coronary steal") were never reported in the prevention trials. The prevention of arterial thrombosis and the positive remodeling of the arterial system (arteriogenesis) by elevated blood flows suggest that dipyridamole may be able to halt the progression of organ manifestations of atherosclerosis. Clinical trials for the secondary prevention of vascular occlusions in other vascular beds should be encouraged.


Efficient display of active Geotrichum sp. lipase on Pichia pastoris cell wall and its application as a whole-cell biocatalyst to enrich EPA and DHA in fish oil.
Geotrichum sp. lipase (GSL) was first displayed on the cell wall of Pichia pastoris on the basis of the a-agglutinin anchor system developed in Saccharomyces cerevisiae . Surface display levels were monitored using Western blotting, immunofluorescence miscroscopy, and fluorescence-activated cell sorting analysis. Lipase activity of the yeast whole cells reached a maximum at 273 ± 2.4 U/g of dry cells toward olive oil after 96 h of culture at 30 °C, with optimal pH and temperature at 7.5 and 45 °C, respectively. Displayed GSL exhibited relatively high stability between pH 6.0 and 8.0 and retained >70% of the maximum activity. The surface-displayed lipase retained 80% of its original activity after incubation at 45 °C for 4 h. Moreover, the GSL-displaying yeast whole cells were then used as a biocatalyst to enrich eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil on the basis of selective hydrolysis. As a result, EPA and DHA increased from 1.53 and 24.1% in the original fish oil to 1.85 and 30.86%, which were increases of 1.21- and 1.29-fold, respectively. The total yield of EPA and DHA reached 46.62%.


Nonadherence to asthma treatment and failure of therapy.
Adherence to clinical regimens is paramount in determining their effectiveness. Asthma is a chronic illness that requires consistent, anti-inflammatory therapy. A "high risk" profile for fatal or near-fatal asthma has emerged that includes suboptimal pharmacotherapy in conjunction with severe disease, psychologic dysfunction, black race or Hispanic-American ethnicity, and residence in an urban area with a high concentration of poverty. Collaboration between physicians and patients is necessary to allow individuals to make choices informed by medical facts and the physician's experience. Objective monitoring of adherence and pulmonary function and acceptance by the clinicians of the National Asthma Education and Prevention Program guidelines to control airway inflammation are essential in order to reduce the unacceptable rates of morbidity and mortality associated with asthma.


Different roles played by periostin splice variants in retinal neovascularization.
Retinal neovascularization (NV) due to retinal ischemia is one of the major causes of vision reduction in patients with different types of retinal diseases although anti-vascular endothelial growth factor (anti-VEGF) therapy can partially reduce the size of the retinal NV. We recently reported that periostin plays an important role in the development of NV and the formation of preretinal fibrovascular membranes, but the role of the splice variants of periostin on retinal NV has not been determined. We examined the expressions of periostin splice variants in the ischemic retinas of a mouse model of oxygen-induced retinal NV. We also studied the function of periostin splice variants on retinal NV using periostin knock out mice, and the effects of anti-periostin antibodies on retinal NV. Our results showed that the expressions of the periostin splice variants were increased in ischemic retinas. The degree of increase of periostin lacking exon 17 was the highest among the periostin splice variants examined. Both genetic ablation of periostin exons 17 and 21 and antibodies for periostin exons 17 and 21 affected preretinal pathological NV. Inhibition of exon 17 of periostin had the greatest effect in reducing preretinal pathological NV. These findings suggest a causal link between periostin splice variants and retinal NV, and an intravitreal injection of antibody for exon 17 and exon 21 of periostin should be considered to inhibit preretinal pathological NV.


What can improve chronic disease care?
Chronic disease care is a central part of geriatrics. The rise in chronic disease draws attention to the need to find better ways to deliver effective care. This essay examines the effectiveness of several strategies for providing better chronic disease care, reviewing the literature and relying on extant reviews whenever possible. The evidence of effectiveness is mixed; a few areas show promise. The case for cost-effectiveness is even less strong. New strategies may be indicated, including more-proactive monitoring of clinical courses and assessing patients' readiness to participate actively in their own care.


Phage display.
Phage display is a powerful method for the selection and evolution of proteins and peptides. Applications include the generation of potent and novel antibodies, the in vitro improvement of protein affinity and function, epitope discovery, the development of leads for vaccine research and the identification of interacting proteins using cDNA libraries.


[Testicular endocrine function in patients with prostatic cancer receiving medical castration by long-term administration of LH-RH agonists].
Six patients with advanced prostatic cancer who had been treated by long-term administration of LH-RH agonistic preparations (Buserelin or Leupron) were tested for their pituitary-testicular endocrine functions. Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), prolactin (PRL), estradiol (E2) and dihydrotestosterone (DHT) were measured consecutively. In all medically castrated patients, serum levels of LH, FSH, T, DHT and E2 were suppressed and particularly serum T levels were below the castration level of 1.0 ng/ml. On the other hand, serum PRL levels were unchanged after the long-term treatment with the agonists. Serum LH and FSH levels failed to respond to LH-RH stimulation after the treatment, whereas serum T responded to stimulation by human chorionic gonadotropin (hCG) to various degrees. It was remarkable that, in 4 out of 6 medically castrated patients treated up to more than 3 years, serum T response levels above 1.0 ng/ml were noted. It is suggested that testicular endocrine function to secrete T and DHT in patients under treatment with long-term LH-RH agonist administration are still preserved in response to hCG stimulation.


Systemic treatment of colorectal cancer in Serbia: what have we done and what can we offer in the new century?
The treatment of patients with metastatic colorectal cancer (mCRC) has changed dramatically over recent years in Serbia. The more optimal use of 5-fluorouracil (5-FU) in association with leucovorin (LV), the development of new drugs such as oxaliplatin and irinotecan and of the oral fluoropyrimidines, such as capecitabine, have increased therapeutic options and to the improved outcome of patients with mCRC. Throughout our 10-years published papers in international journals, we presented development of chemotherapy for mCRC and improvement in treatment outcome in Serbia. It is shown that combination therapy with 5-FU/LV and oxaliplatin or irinotecan is more active than 5-FU/LV in first line treatment of mCRC. Sequential therapy with FOLFIRI+FOLFOX was the most efficacious combination in comparison to any other 2 drugs combinations. The combination protocols in second line were superior to mono irinotecan and equal to LV5FU2 in terms of time to progression. The oral fluoropyrimidines seems to have an activity comparable to that of i.v. 5-FU/LV. New agents acting on novel targets are under development. Angiogenesis inhibitors, epidermal growth factor inhibitors, COX-2 inhibitors and farnesyl transferase inhibitors might play a role in the future in the treatment of CRC. We will present our first experience with bevacizumab, vascular endothelial growth factor inhibitor.


Metabolomics in Human Acute-Exercise Trials: Study Design and Preparation.
Metabolomics can be of great value in the study of exercise metabolism. However, because of the high intraindividual and interindividual biological variability of the human metabolome, special considerations should be taken into account when designing an acute-exercise metabolomic study. To study different exercise parameters, e.g., different exercise modes, intensities, etc., a crossover study design, where each participant acts as their own control, is preferable to a parallel design, one involving different groups of participants. Moreover, the study should include a no exercise, control trial. Before each trial, participants should follow carefully designed preparatory steps to control for possible confounding factors, i.e., maintain repeatable and constant conditions for all individual trials of the study to minimize variation due to factors other than the one(s) being studied. This chapter focuses on the design of human metabolomic studies, where the intervention is an acute metabolic challenge, such as an exercise bout or a test meal, and presents some basic steps for screening potential participants, performing preliminary tests, preparing for the trial day, and performing the trial.


Reactivation of broccoli peroxidases: structural changes of partially denatured isoenzymes.
Structural changes involved in the reactivation of peroxidases (PODs) from broccoli and horseradish (HRP) following heat denaturation were investigated by using circular dichroism and absorption spectroscopy. Cooling heat-treated enzymes resulted in rapid refolding of the secondary structure into an inactive structural species, similar in conformation to the native enzyme. Reassociation of heme to the refolded peroxidase, as well as molecular rearrangement of the structure around the heme, occurs during incubation at approximately 25 degrees C and results in the return of biological activity. The secondary structure of neutral broccoli POD (N) is relatively heat labile, resulting in a rapid loss of activity, but the level of reactivation is high because the structure at the heme pocket is relatively stable. Acidic broccoli POD and HRP are more heat stable than N, but have a low degree of reactivation. Loss of activity is due primarily to alteration of the structure at the heme pocket. Effects of bovine serum albumin and pH on reactivation of PODs are also discussed. Peroxidase; reactivation; horseradish; broccoli; circular dichroism; absorption spectroscopy.