CD4-independent protective cytotoxic T cells induced in early life by a non-replicative delivery system based on virus-like particles

The relative immaturity of the neonatal immune system limits CD4(+) Th1 and cytotoxic T lymphocyte (CTL) responses, and represents a significant challenge for the development of vaccines against intracellular pathogens. In this report, we demonstrate the ability of a non-replicative delivery system based on parvovirus-like particles (VLP) to induce CTL responses in the neonatal period. A single immunization of 1-week-old BALB/c mice with recombinant VLP carrying a CD8(+) T cell determinant from lymphocytic choriomeningitis virus (VLP-LCMV) induced antigen-specific CD8(+) cytotoxic T cells that were similar to those elicited by adult immunization, as assessed by cytotoxic activity, interferon (IFN)-gamma secretion, cytotoxic precursor cell frequencies, in vitro avidity for antigen and protective activity against viral challenge. These CTL responses are elicited within 2 weeks of a single immunization, in the absence of adjuvant and independently of the presence and help of CD4(+) T cells, highlighting the potential of VLP as candidate vaccine vectors in early life.     

A comparison of rational versus empirical methods in the prediction of psychotherapy outcome

Recent studies have shown that providing feedback to therapists based on comparing their clients' progress to a set of rational, clinically derived algorithms delineating various categories of progress has enhanced outcomes for clients predicted to show poor treatment outcomes (signal‐alarms). One prior study indicated that an empirically derived method more accurately predicts outcome than the rational method. The present study replicated the comparison of empirical and rational methods, while adding an additional layer of effect size analyses to further clarify predictive accuracy. The two methods were approximately equivalent in their accurate detection of cases that had a final negative outcome. However, the rational method had significantly lower overall predictive accuracy due to its high percentage of false negative predictions. Further, progressively more optimistic predictions based on the empirical method were associated with greater improvement for the average client. This was not the case for the rational method. Copyright © 2006 John Wiley & Sons, Ltd.     

Differentiation of Peptococcus and Peptostreptococcus by gas-liquid chromatography of cellular fatty acids and metabolic products

Gas-liquid chromatographic (GLC) profiles of cellular fatty acids and metabolic products were useful in identifying strains of Peptococcus saccharolyticus, Peptococcus asaccharolyticus, Peptostreptococcus anaerobius, Peptostreptococcus micros, and Streptococcus intermedius. The GLC results supported the recent taxonomic decision to transfer aerotolerant Peptostreptococcus species to the genus Streptococcus. Because inconsistencies in the results prevented our differentiating Peptococcus prevotii. Peptococcus magnus, and Peptococcus variabilis by GLC, additional strains will have to been examined. These GLC techniques are amenable to routine use; however, for interlaboratory results to be meaningful, the classification and nomenclature of the anaerobic gram-positive cocci should be standardized.     

Differences in assembly or stability of complex I and other mitochondrial OXPHOS complexes in inherited complex I deficiency

NADH-ubiquinone oxidoreductase (complex I) deficiency is amongst the most encountered defects of the mitochondrial oxidative phosphorylation (OXPHOS) system and is associated with a wide variety of clinical signs and symptoms. Mutations in complex I nuclear structural genes are the most common cause of isolated complex I enzyme deficiencies. The cell biological consequences of such mutations are poorly understood. In this paper we have used blue native electrophoresis in order to study how different nuclear mutations affect the integrity of mitochondrial OXPHOS complexes in fibroblasts from 15 complex I-deficient patients. Our results show an important decrease in the levels of intact complex I in patients harboring mutations in nuclear-encoded complex I subunits, indicating that complex I assembly and/or stability is compromised. Different patterns of low molecular weight subcomplexes are present in these patients, suggesting that the formation of the peripheral arm is affected at an early assembly stage. Mutations in complex I genes can also affect the stability of other mitochondrial complexes, with a specific decrease of fully-assembled complex III in patients with mutations in NDUFS2 and NDUFS4. We have extended this analysis to patients with an isolated complex I deficiency in which no mutations in structural subunits have been found. In this group, we can discriminate between complex I assembly and catalytic defects attending to the fact whether there is a correlation between assembly/activity levels or not. This will help us to point more selectively to candidate genes for pathogenic mutations that could lead to an isolated complex I defect.     

Gill repartition of two Monogenea: Diplectanum aequans (Wagener 1857), Diesing 1958 and D. laubieri Lambert and Maillard 1974 (Monogenea, Monopisthocotylea) simultaneous parasites of the sea bass Dicentrarchus labrax (author's transl)]

The close observation of the gill habitat of two Monogeneans Diplectanum aequans (Wagener 1857) Diesing 1858 and Diplectanum laubieri Lambert and Maillard 1974 shows the microbiotopes of these two simultaneous parasites of the Sea-bass: Dicentrarchus labrax. The authors propose to study the distribution of the Monogeneans with references to three theoretic axes in the gill chambers. For the Diplectanum, the populations spread on each of these axes according to preferential gradients.     

Interference of oral immunization with the intestinal absorption of heterologous albumin

Rats were immunized with human serum albumin (HSA) by a single intragastric administration of 200 mg of HSA. Two weeks later their capacity to absorb a similar intragastric test dose of HSA was found to be greatly impaired, the concentrations of HSA in mesenteric venous serum having been reduced to 33 %, 25 % and 60 % of those in similarly tested but unprimed animals, respectively 1, 2 and 3 hours after the intragastric test dose. Rats given antigen (HSA) together with intestinal secretions from intragastric immunized rats also showed a striking decrease in intestinal absorption of a test dose, compared to controls given the antigen together with nonimmune intestinal secretions. From these data it is concluded that local immunization of the gut impairs its capacity to absorb the corresponding antigen, and that this effect is largely due to the failure to absorb antigen bound to secretory antibodies.     

The mutational specificity of l-nitroso-6-nitropyrene in the lacI gene of Escherichia coli strains deficient in nucleotide excision repair

We have examined the mutational specificity of 1-nitroso-6-nitropyrene(1,6-NONP), an activated metabolite of the carcinogen 1,6-dinitropyrene,in the lacI gene of Escherichia coli strains which are deficientin nucleotide excision repair (strain NR6113,     

Stimulation of cranial vessels excites nociceptive neurones in several thalamic nuclei of the cat

Summary Extracellular recordings were made in the thalamus of cats anaesthetized with chloralose and urethane following electrical, mechanical and chemical stimulation of the superior sagittal sinus or middle meningeal artery. Facial receptive fields were looked for using electrical and mechanical stimuli. The locations of fifty-six cells were verified histologically. Twenty six cells were located in the ventroposteromedial nucleus (VPM) and six in its ventral periphery (VPMvp). All units in VPM had facial receptive fields, usually involving the first trigeminal division. Cells with nociceptive receptive fields or responding to the craniovascular application of bradykinin were often found in the periphery or “shell” region of VPM. Other craniovascular nociceptive cells were found in VPMvp, in the posterior group and in the intralaminar complex. This study shows that craniovascular afferents in the cat project to several thalamic nuclei and implicate VPM especially in craniovascular nociception.     

Simple DNA extraction method for dried blood spots and comparison of two PCR assays for diagnosis of vertical human immunodeficiency virus type 1 transmission in Rwanda

Dried blood spots (DBS) on filter paper facilitate the collection, transport, and storage of blood samples for laboratory use. A rapid and simple DNA extraction procedure from DBS was developed and evaluated for the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection in children by an in-house nested-PCR assay on three genome regions and by the Amplicor HIV-1 DNA prototype assay version 1.5 (Roche Molecular Systems). A total of 150 samples from children born to HIV-1-infected mothers were collected in Kigali, Rwanda, in parallel as DBS and as peripheral blood mononuclear cell (PBMC) pellets. The results obtained on DBS by the two PCR assays were compared to the results of nested PCR on PBMCs. Of 150 PBMC samples, 10 were positive, 117 were negative, and 23 were indeterminate for HIV-1 infection. In DNA extracted from filter papers and amplified by using the in-house nested PCR, 9 of these 10 positive samples (90%) were found to be positive, and 1 was found to be indeterminate (only the pol region could be amplified). All of the negative samples and all of the 23 indeterminate samples tested negative for HIV-1 infection. When we used the Amplicor DNA test on DBS, all of the 10 PBMC-positive samples were found to be positive and all of the 23 indeterminate samples were found to be negative. Of the PBMC-negative samples, 115 were found to be negative and 2 were found to be indeterminate. We conclude that this simple rapid DNA extraction method on DBS in combination with both detection methods gave a reliable molecular diagnosis of HIV-1 infection in children born to HIV-infected mothers.