Nuclear medicine technologists are able to accurately determine when a myocardial perfusion rest study is necessary

ABSTRACT: BACKGROUND: In myocardial perfusion scintigraphy (MPS), typically a stress and a rest study is performed. If the stress study is considered normal, there is no need for a subsequent rest study. The aim of the study was to determine whether nuclear medicine technologists are able to assess the necessity of a rest study. METHODS: Gated MPS using a 2-day 99mTc protocol for 121 consecutive patients were studied. Visual interpretation by 3 physicians was used as gold standard for determining the need for a rest study based on the stress images. All nuclear medicine technologists performing MPS had to review 82 training cases of stress MPS images with comments regarding the need for rest studies, and thereafter a test consisting of 20 stress MPS images. After passing this test, the nuclear medicine technologists in charge of a stress MPS study assessed whether a rest study was needed or not or if he/she was uncertain and wanted to consult a physician. After that, the physician in charge interpreted the images and decided whether a rest study was required or not. RESULTS: The nuclear medicine technologists and the physicians in clinical routine agreed in 103 of the 107 cases (96%) for which the technologists felt certain regarding the need for a rest study. In the remaining 14 cases the technologists were uncertain, i.e. wanted to consult a physician. The agreement between the technologists and the physicians in clinical routine was very good, resulting in a kappa value of 0.92. There was no statistically significant difference in the evaluations made by technicians and physicians (P=0.617). CONCLUSIONS: The nuclear medicine technologists were able to accurately determine whether a rest study was necessary. There was very good agreement between nuclear medicine technologists and physicians in the assessment of the need for a rest study. If the technologists can make this decision, the effectiveness of the nuclear medicine department will improve.     

How to deal with the early GWAS data when imputing and combining different arrays is necessary

Genotype imputation has become an essential tool in the analysis of genome-wide association scans. This technique allows investigators to test association at ungenotyped genetic markers, and to combine results across studies that rely on different genotyping platforms. In addition, imputation is used within long-running studies to reuse genotypes produced across generations of platforms. Typically, genotypes of controls are reused and cases are genotyped on more novel platforms yielding a case-control study that is not matched for genotyping platforms. In this study, we scrutinize such a situation and validate GWAS results by actually retyping top-ranking SNPs with the Sequenom MassArray platform. We discuss the needed quality controls (QCs). In doing so, we report a considerable discrepancy between the results from imputed and retyped data when applying recommended QCs from the literature. These discrepancies appear to be caused by extrapolating differences between arrays by the process of imputation. To avoid false positive results, we recommend that more stringent QCs should be applied. We also advocate reporting the imputation quality measure (R(T)(2)) for the post-imputation QCs in publications.     

Why offense is reduced when rats are tested in a strange cage

Tests of isolation-induced fighting and competitive fighting of rats in various conditions were used to evaluate the hypothesis that there is more offense in a familiar than in a strange cage because of the operation of an “olfactory comparator.” The hypothesis was rejected. Although there was more offense in a familiar than in a strange cage, there was not, as predicted, more offense when Petri dishes containing familiar scent-markings were put in the strange test cage, and there was not, as predicted, a difference in the effect between isolation-induced fighting and competitive fighting. The data were consistent with either one or both of two alternative hypotheses: 1) that the strange cage produces fear (neophobia) that reduces offense; or 2) that the strange cage activates exploratory and scent-marking behaviors that compete with offense. The first of these two alternative hypotheses was also consistent with a finding that handling the test animal prior to a test reduces subsequent offense.     

Late dendritic cells are still able to evoke a potent alloreactive CTL response

On exposure to maturation stimuli, immature dendritic cells (DCs) undergo changes that turn them into potent amplifiers of innate immunity and into antigen-presenting cells (APCs) able to prime na?ve T cells. However, their progression through the maturation process is very rapid and finally ends in apoptosis. The aim of our study was to investigate the importance of the maturation stage of DCs, defined by morphology, expression of surface markers and IL-12 production, for their immunostimulatory capacity. DCs were matured with LPS, monocyte-conditioned medium (MCM) or TNF-alpha, sampled several times during a 3-day long maturation period and used as stimulators of allogeneic T cells over a wide range of DC/T cell ratios. T-cell response was assessed by cell proliferation, CTL generation and IFN-gamma production. Our results indicate that the in vitro T cell response is determined mainly by the level of expression of co-stimulatory molecules on DCs and the DC/T cell ratio in the culture. Thus, DCs matured for over 20h, with high expression of co-stimulatory molecules, can still induce a potent CTL response at DC/T cell ratios of 1:10 and 1:20, although their IL-12 production, as well as their ability to induce IFN-gamma production by T cells, are both decreased. In contrast, the CTL response at DC/T cell ratios of 1:2 and 1:5 can be profoundly decreased. Notably, the proportion of proliferating CD4+ T cells in these cultures is reduced. This could well be the reason for the absence of CTL response, since we showed that, even in the case of high expression of co-stimulatory molecules on DCs, generation of CTLs still depends on CD4+ T cells. Our study emphasizes the importance of strong expression of co-stimulatory molecules on DCs and of their ability to activate CD8+ and CD4+ T cells concomitantly in order to initiate a potent cell-mediated immune response. We therefore suggest that a combination of early DCs, which are strong producers of cytokines, and late DCs, which have high expression of co-stimulatory molecules, could prove beneficial in the attempt to initiate in vitro and in vivo cell-mediated immune responses for therapeutic purposes.     

Scaling is necessary when making comparisons between shapes of event-related potential topographies: a reply to Haig et al.

A. R. Haig, E. Gordon, and S. Hook (1997) disputed G. McCarthy and C. C. Wood's (1985) contention that scaling should be used when assessing the statistical significance of between condition (or group) differences in the shapes of event-related potential (ERP) scalp topographies. Haig et al. based their contention upon the lack of empirical realism in McCarthy and Wood's model of within-group ERP noise, claiming that McCarthy and Wood's results could not be generalized to realistic ERP data. We argue, on both empirical and theoretical grounds, that Haig et al. do not make a compelling case against generalization of McCarthy and Wood's results. Moreover, Haig et al.'s conclusion is based upon a misconception of how scaling should be used. We conclude that when a quantitative measure of differences between topographic shapes is needed, scaling is not an option--it is a requirement.     

Locomotion through apertures when wider space for locomotion is necessary: adaptation to artificially altered bodily states

The objective of this study is to describe the adaptability of the central nervous system to safely cross a narrow aperture when the space required for passage is transiently extended with external objects under different locomotor constraints. In one of four locomotion forms (normal walking, walking while holding a 63-cm horizontal bar with or without rotating the shoulders to cross a door opening, and wheelchair use), nine participants were asked to pass through an aperture created by two doors (the relative aperture widths were 1.02, 1.10, and 1.20 times their maximum horizontal dimension under each form of locomotion) without a collision. The kinematic analyses showed that, when the participants rotated their shoulders while walking and holding a bar, virtually the same locomotor patterns as those during normal walking were observed: shoulder rotation was regulated well in response to the width of an aperture, and no collisions occurred. When shoulder rotations were restricted while walking and holding a bar or using a wheelchair, a large reduction in the speed of movement was observed as the participants approached the door, and, furthermore, the modulation in speed was dependent on the width of the aperture. In addition, the participants crossed at the center of aperture more accurately; nevertheless, collision sometimes occurred (more frequently, during wheelchair use). These findings reveal that movement constraints on shoulder rotation are likely to be a critical factor in determining whether quick and successful adaptation takes place.     

Purification of a factor from human peritoneal fluid that is able to immobilize spermatozoa

Human peritoneal fluid has been claimed to influence sperm motility.This report gives evidence for the presence in mid-cycle peritonealfluid of a protein-bound, lipidic (hydrophobic) component ableto immobilize spermatozoa as a function of time. This componentwas extracted from molecular weight-sieving and ion-exchange/highpressure liquid chromatography (HPLC)-purified peritoneal fluidfractions by either chloroform/methanol or charcoal treatments;resuspension of the chloroform/methanol extract with BWW-bufferand subsequent testing on spermatozoa resulted in sperm immobilization.Sequential or step-down chromatographic procedures (molecularweight-sievingcation-exchangeanion-exchange HPLC separationsof native peritoneal fluid) and extensive dialysis against doubledistilled water allowed the purification of the sperm immobilizingfactor, as evidenced by the shorter incubation times necessaryfor sperm immobilization. Furthermore, the active fraction wasfound to immobilize spermatozoa without affecting its viability.Separation of the chloroform/methanol extracted immobilizingfraction on thin layer chromatography under conditions for phospholipiddetection allowed the identification of a characteristic bandwhich, after re-extraction, was found to be the sperm immobilizingsubstance. This factor does not contain choline, ethanolamineor serine. These results suggest that some lipidic peritonealfluid components may influence sperm motility.     

Plasmodium falciparum is able to invade erythrocytes through a trypsin-resistant pathway independent of glycophorin B

Plasmodium falciparum invades erythrocytes through multiple ligand-receptor interactions, with redundancies in each pathway. One such alternate pathway is the trypsin-resistant pathway that enables P. falciparum to invade trypsin-treated erythrocytes. Previous studies have shown that this trypsin-resistant pathway is dependent on glycophorin B, as P. falciparum strains invade trypsin-digested glycophorin B-deficient erythrocytes at a highly reduced efficiency. Furthermore, in a recent study, the P. falciparum 7G8 strain did not invade glycophorin B-deficient erythrocytes, a finding that was not confirmed in the present study. To analyze the degree of dependence on glycophorin B for invasion by P. falciparum through the trypsin-resistant pathway, we have studied the invasion phenotypes of five parasite strains, 3D7, HB3, Dd2, 7G8, and Indochina I, on trypsin-treated normal and glycophorin B-deficient erythrocytes. Invasion was variably reduced in glycophorin B-deficient erythrocytes. Four strains, 3D7, HB3, Dd2, and Indochina I, invaded trypsin-treated erythrocytes, while invasion by the 7G8 strain was reduced by 90%. Among the four strains, invasion by 3D7, HB3, and Dd2 of trypsin-digested glycophorin B-deficient erythrocytes was further reduced. However, Indochina I invaded trypsin-digested glycophorin B-deficient erythrocytes at the same efficiency as its invasion of trypsin-digested normal erythrocytes. This strongly suggests that the Indochina I strain of P. falciparum is not dependent on glycophorin B to invade through a trypsin-resistant pathway as are the strains 3D7, HB3, and Dd2. Thus, P. falciparum is able to invade erythrocytes through a glycophorin B-independent, trypsin-resistant pathway.     

B-1 lymphocytes are able to produce IL-10, but is not pathogenic during Leishmania (Leishmania) amazonensis infection

Over the years research has found an association between B lymphocytes and pathogenesis during Leishmania sp. infections. Recently we demonstrated that B-2 lymphocytes are the main producers of IL-10 during L. amazonensis infection, and that the disease severity in BALB/c mice was attributed to these IL-10-producing B-2 lymphocytes. Here, we aim to understand the role of peritoneal B-1 lymphocytes in the pathogenesis of L. amazonensis infection. We found that infection resulted in a decrease in the number of B-1a lymphocytes and increase in B-1b lymphocytes in the peritoneal cavity of WT BALB/c mice but not in B lymphocyte deficient mice (BALB/Xid) mice. In vitro interaction between B-1 lymphocytes and L. amazonensis showed that the amastigote form of the parasite was able to induce higher levels of IL-10 in B-1 lymphocytes derived from infected BALB/c mice than the promastigote. Moreover, B-1 lymphocytes derived from infected mice produced more IL-10 than B-1 lymphocytes derived from naïve mice under amastigote interaction. However, the repopulation of BALB/Xid mice with B-1 lymphocytes from WT BALB/c mice did not affect the lesion development. Together, these results suggest that although B-1 lymphocytes are able to produce IL-10 during in vitro interaction with L. amazonensis, they are not directly related to pathogenesis in vivo.     

Melatonin and CAPE are able to prevent the liver from oxidative damage in rats: an ultrastructural and biochemical study

The liver continuously produces free radicals and reactive oxygen species (ROS) as part of metabolic process. These free radicals are neutralized by an elaborate antioxidant defense system consisting of enzymes and numerous nonenzymatic antioxidants like flavonoids. In this study, we have evaluated effects of melatonin and caffeic acid phenethyl ester (CAPE) to young and aged rat liver. Aging-related hepatic changes examined by light and electron microscopy and biochemical methods. Melatonin and CAPE decreased tissue malondialdehyde (MDA) levels in aged rats. Melatonin elevated tissue glutathione peroxidase (GSH-Px) activity and tGSH level, whereas CAPE elevated tissue catalase activity in aged rats. This study demonstrates that both melatonin and CAPE are beneficial in delaying age-related hepatocellular changes. Melatonin and CAPE supplementation in older ages may support liver to protect itself from various damaging agents including infectious agents and toxins.     

After ivermectin, is it necessary to look again for a macrofilaricide?

The use of ivermectin for mass treatment in human onchocerciasis is justified, because no other drug was available. But, as this compound is ineffective on adult filariae and as the emergence of drug resistance is quite possible, the evaluation of new chemical compounds for their macrofilaricidal activity should be continued. Several new compounds of interest are presented.     

A synthetic mimetic of CD4 is able to suppress disease in a rodent model of immune colitis

CD4⁺ mucosal T cells mediate the intestinal inflammation in Crohn's disease and may serve as an important target for immune intervention. Here we assessed the therapeutic effect of a synthetic mimetic of CD4 designed to mimic both the sequence and conformation of the complementarity-determining region 3 of murine CD4 V1 domain (rD-mPGPtide) in a mouse colitis model using immunization with 2,4,6-trinitrobenzene sulfonic acid (TNB). i. v. administration of the rD-mPGPtide but not control scrambled peptide could suppress severe inflammation in the chronic colitis mouse model. After treatment with the rD-mPGPtide, a striking improvement of diarrhea and acute wasting disease was observed with decreased mortality. Serum anti-TNB antibody titers, CD45RB^lowCD4⁺ T cells in the lamina propria and IFN-γ mRNA expression in the mucosa were significantly decreased with the rD-mPGPtide treatment. Anti-CD4 antibody also suppressed disease by depletion of CD45RB^highCD4⁺ T cells in the colonic mucosa. The observation that the synthetically engineered analogue of murine CD4 inhibits inflammation in a rodent disease model by different mechanisms than anti-CD4 antibody suggests that a human version of this peptide has potential therapeutic utility in CD4⁺ mucosal T cell-mediated intestinal inflammation in Crohn's disease.     

Oral sensitization to peanut is highly enhanced by application of peanut extracts to intact skin, but is prevented when CpG and cholera toxin are added

BACKGROUND: CpG oligonucleotides might offer an alternative to conventional immunotherapy in preventing and potentially reversing Th2-biased immune deregulation which leads to allergy. However, non-invasive ways of administration, especially in peanut-allergic patients, should be explored. METHODS: One hundred micrograms of whole peanut protein extract (PE) alone, or mixed with cholera toxin (CT, 50 microg) plus CpG (100 microg) as adjuvant, was applied on intact skin of mice (40 min, twice). Initiation of an immune response was monitored by detection of specific antibodies in sera. The effect of this pretreatment on a further oral sensitization by PE was then evaluated by assaying antibodies and cytokines specific for PE and purified allergens. Cytokine production in liver 40 min after skin application was also assayed. RESULTS: Two brief skin applications of PE alone highly potentiated further oral sensitization, as demonstrated by very intense specific IgE, IL-4 and IL-5 productions. Conversely, skin pretreatment with PE and CT + CpG efficiently prevented further sensitization via gastro-intestinal exposure. In both cases, the specificity of the antibodies and cytokines was the same as in control mice. CT + CpG treatment allowed the rapid production of IL-12 and TGFbeta in liver and of specific IgG2a in sera, suggesting the activation of Th1 and/or regulatory T cells. CONCLUSIONS: Oral sensitization to peanut is highly enhanced by a previous short exposure of allergens to intact skin. Conversely, the use of CT + CpG adjuvant for skin application efficiently prevents further oral sensitization. The potential of such treatment in specific immunotherapy needs to be evaluated.