Detoxified Haemophilus ducreyi cytolethal distending toxin and induction of toxin specific antibodies in the genital tract

Haemophilus ducreyi causes genital ulceration (chancroid), a sexually transmitted infection and still an important factor which contributes to the spread of HIV in developing countries. The bacterium produces a cytolethal distending toxin (HdCDT) causing cell cycle arrest and apoptosis/necrosis of human cells and contributes to the aggravation of ulcers. The aim of the study was to induce toxin-neutralizing antibodies in the genital tract of mice. Repeated subcutaneous (sc) immunisations with 5–10 μg active HdCDT induced low levels of serum anti-HdCDT IgG without neutralizing capacity. High levels of specific IgG1 antibodies in serum and genital tract were generated after sc immunisations with 10 μg formaldehyde detoxified HdCDT toxoid alone and the addition of aluminium salts or RIBI (based on the lipid A moiety) as adjuvant further increased the level of serum antibodies. A high correlation was found between elevated levels of anti-HdCDT IgG in sera, the level of neutralizing activity and the antibody level in genital tract (r = 0.8). Thus, induction of high antibody levels specific to HdCDT in the genital tissue can be achieved by parenteral immunisation with the toxoid. The HdCDT toxoid can be considered as a candidate component in vaccine against chancroid.     

Effect of reserpine on the brain uptake of carbon 11 methamphetamine and its N-propagyl derivative, deprenyl

The enantiomers of methamphetamine (MAMP) and its N-propagyl derivative, deprenyl, were labelled with carbon 11, and the tissue distribution of these labelled compounds in mice was studied. Both enantiomers of 11C-MAMP rapidly entered into the brain and then disappeared according to a single exponential curve. The enantiomers of 11C-deprenyl were also rapidly distributed to various organs in the same manner. With regard to elimination, however, a stereoselective, long-term retention of radioactivity in the brain, heart and lung, due to its irreversible binding with monoamine oxidase B, was observed for L-11C-deprenyl. In reserpinized mice, the initial brain uptake of both the L and D forms of 11C-MAMP was significantly decreased. On the other hand, the brain uptake of both enantiomers of 11C-deprenyl was slightly increased by pretreatment with reserpine. A significant and non-stereoselective elevation of the lung uptake of 11C-deprenyl was also seen in reserpinized mice. In addition, both the relative tissue distribution and ratios of radioactivity in the brain compared with blood or heart at 1 and 5 min after the injection of 11C-labelled methanol in mice were not changed by reserpine. These results indicate that the transport or binding processes of these amines rather than the blood flow might be altered by reserpine. There would be an important role of the pKa values of amines in both processes. The reduction of brain uptake as well as the change in ratio between brain and heart of L-11C-MAMP in reserpinized mice 1 min after injection were reversed by treatment with amphetamine in a dose-related manner.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of Bz-DOTA and CHX-A″-DTPA on the biodistribution of ABD-fused anti-HER2 Affibody molecules: implications for <Superscript>114m</Superscript>In-mediated targeting therapy

Purpose  Affibody molecules represent a novel class of high-affinity agents for radionuclide tumour targeting. Fusion of the Affibody molecules with an albumin-binding domain (ABD) enables modification of the blood kinetics of the Affibody molecules and reduction of the renal dose. ¹⁷⁷Lu-CHX-A″-DTPA-ABD-(Z_HER2:342)₂, an anti-HER2 Affibody molecule-ABD fusion protein has earlier demonstrated promising results in treatment of HER2-expressing micro-xenografts in mice. The use of the in vivo generator ^114mIn/¹¹⁴In as a label for ABD-fused Affibody molecules would create preconditions for efficient treatment of both micrometastases (due to conversion and Auger electrons of ^114mIn) and bulky tumours (due to high-energy beta particles from the daughter nuclide ¹¹⁴In). The goal of this study was to investigate if different chelators influence the biodistribution of ABD-(Z_HER2:342)₂ and to find an optimal chelator for attachment of ^114mIn to the Affibody molecule-ABD fusion protein. Methods  Isothiocyanate derivatives of Bz-DOTA and CHX-A″-DTPA were coupled to ABD-(Z_HER2:342)₂. The cellular processing of both conjugates was studied in vitro. The influence of chelators on the biodistribution was investigated in mice using double isotope (^114mIn and ¹¹¹In) labelling. Results  The apparent affinity of CHX-A″-DTPA-ABD-(Z_HER2:342)₂ and Bz-DOTA-ABD-(Z_HER2:342)₂ to the extracellular domain of HER2 was similar, 13.5 and 15.0 pM, respectively. It was found that both conjugates were internalized by SKOV-3 cells. The use of CHX-A″-DTPA provided better cellular retention of the radioactivity, better tumour accumulation of radioactivity and better tumour to organ dose ratios than Bz-DOTA-ABD-(Z_HER2:342)₂. Conclusion  CHX-A″-DTPA is more suitable for ^114mIn labelling of Affibody molecule-ABD fusion proteins for radionuclide therapy.     

Serum beta2-microglobulin in women with breast cancer and in age-matched, non-hospitalized controls.

Recently, reports have shown an increase of β₂-microglobulin in malignant disorders of different types including breast cancer. The present study was undertaken to evaluate further serum levels of β₂-microglobulin in 129 patients with breast cancer at various stages.As serum β₂-microglobulin concentration is significantly correlated to age, age-matched controls were used for comparisons.No significant difference was found in serum β₂-microglobulin levels in patients with breast cancer compared to age-matched controls, even when the levels were higher than reported for healthy individuals. The β₂-microglobulin concentration was not correlated to the TNM classification of the tumour.In order to compensate for the influence of renal function on serum β₂-microglobulin levels, the quotient between β₂-microglobulin and creatinine was calculated in 93 patients. The quotient was neither related to the stage of the disease nor to the tumour diameter. In 16 patients β₂-microglobulin was determined at different times in relation to mastectomy. There were no significant changes in circulating β₂-microglobulin levels after removal of the primary tumour.The present study underlines the need for age-matched controls, since serum β₂-microglobulin increases with age. The reason for such an increase could however only be speculated on, but may be explained by the high frequency of diseases causing known elevation in serum β₂-microglobulin such as infections of inflammatory disorders, liver diseases or renal dysfunction. Further studies must be undertaken in other malignancies before the role of serum β₂-microglobulin determination in such disorders can be stated.     

Nonviral and viral gene transfer into different subsets of human dendritic cells yield comparable efficiency of transfection

Among the many promising cancer immunotherapeutic strategies, dendritic cells (DC) have become of particular interest. This study aims to optimize a clinical grade protocol for culture and transfection of human DC. Monocytes and CD34(+) hematopoietic stem cells (HSC) from same donor were differentiated under serum-free conditions and analyzed for their susceptibility to several recently described nonviral transfection methods as compared with established virally mediated gene transfer. Nonviral gene transfer methods studied were square-wave electroporation, lipofection, and particle-mediated transfer of plasmid DNA or in vitro transcribed mRNA. We conclude that DNA is not suitable for transduction of DC using nonviral methods. In contrast, mRNA and square-wave electroporation reproducibly yields 60% and 50% transfected monocyte- and CD34(+)-derived DC, respectively, measured at protein level, without affecting the cell viability. Thus, the transfection efficiency of this method is comparable with the 40-90% transgene expression obtained using retroviral (RV) or adenoviral (AdV) vectors in CD34(+)- and monocyte-derived DC, respectively. In monocyte-derived DC, however, the amount of protein expressed per-cell basis was higher after AdV (MOI = 1000) compared with mRNA electroporation-mediated transfer. This is the first study directly demonstrating side-by-side that mRNA electroporation into DC of different origin indeed results in a comparable number of transduced cells as when using virus-mediated gene transfer.     

Ratio between serum IL-8 and pepsinogen A/C: a marker for atrophic body gastritis

BACKGROUND AND AIMS: Elevated serum gastrin and a low pepsinogen A/C ratio are well-recognized markers for atrophic body gastritis (ABG). We have shown that the presence of body atrophy is also associated with elevated serum pro-inflammatory cytokines. This study tested the hypothesis that serum cytokines provide additional information to gastrin and pepsinogens in screening for ABG. METHODS: Two hundred and twenty-six consecutive patients were investigated on referral for upper gastrointestinal endoscopy: 150 were patients with gastro-oesophageal reflux disease, receiving acid inhibitory medication either with proton pump inhibitors (n = 113) or with histamine2-receptor antagonists (n = 37), and 76 were nontreated controls, who had normal endoscopic findings. Gastric mucosal biopsies were sampled for histological examination (Sydney classification). Serum samples were analyzed for gastrin, chromogranin A (CgA), and pepsinogens A and C by RIA, and for the interleukins (IL)-1beta, IL-6, and IL-8 by ELISA. RESULTS: Subjects with ABG had significantly higher serum gastrin (P < 0.01) and serum CgA (P < 0.01) levels and significantly lower pepsinogen A/C ratios (P < 0.001) than those without ABG. Additionally, serum IL-1beta, IL-6 and, especially, IL-8 levels were significantly higher in the subjects with than in those without ABG (P < 0.0001, for all cytokines). To optimize the detection of body atrophy we defined the ABG index: the ratio between the simultaneously measured IL-8 and pepsinogen A/C. The area under the ROC curve for the ABG index was significantly greater than that for serum gastrin and for serum pepsinogen A/C alone (0.91 +/- 0.029 vs. 0.72 +/- 0.042, and vs. 0.83 +/- 0.031, P = 0.018 and P = 0.049). Using the ABG index at a cut-off value of 1.8 pg mL-1, 91% of the cases were classified correctly. CONCLUSIONS: The ratio between serum IL-8 and pepsinogen A/C accurately predicts the presence of ABG. We therefore propose the ABG index as a noninvasive screening test for ABG in population-based studies.     

Mature dendritic cells induce tumor-specific type 1 regulatory T cells

The aim of this study was to compare the tumor antigen-specific T-cell repertoire generated by transduced, human dendritic cells (DCs). The transductions were three commonly used antigen delivery procedures: adenovirus (AdV) infection, RNA electroporation, and liposome-mediated protein transfection. The DCs in each experimental group were transfected with similar efficacy and matured using TNF-alpha, anti-CD40, or lipopolysaccharide. Regardless of the gene transfer method or the maturation stimuli used, the DCs were indistinguishable with regard to surface phenotype and allostimulatory capacity. With the exception of the Adv transduced group, the T cells generated were tumor antigen specific, as characterized by high IFN-gamma production. The T cells generated upon stimulation with DCs subjected to AdV infection, and subsequently treated with TNF-alpha, exhibited tumor antigen specificity, but accompanied by reduced proliferation and IFNgamma production and increased IL-10 production. Moreover, these T cells exerted a suppressive effect on both autologous and allogeneic lymphocytes resembling type 1 regulatory T cells (Tr1). The authors show that mature DCs may induce tumor antigen-specific Tr1 cells by the appearance of high IL-10 and low IL-12. Similar results were also obtained with AdV-infected and TNF-matured DCs regardless of the transgene used. This work supports the conclusion that it can no longer be assumed that mature DCs induce only antitumor reactive T cells.     

Predictive Association of Smoking with Depressive Symptoms: a Longitudinal Study of Adolescent Twins

Prevention Science - Longitudinal, genetically informative studies of the association between cigarette smoking and depressive symptoms among adolescents are limited. We examined the longitudinal...     

Impaired executive performance in healthy siblings of schizophrenia patients in a population-based study

There is increasing evidence that healthy siblings of schizophrenia patients have similar, although milder, neuropsychological deficits than their affected family members. However, the interpretation of these findings has been complicated by methodological differences, for example the selection of relatives studied and the sensitivity of tests used. We studied neuropsychological functioning in schizophrenia families in representative, population-based samples of schizophrenia patients (n=81) and healthy siblings (n=78) from 58 families, and control subjects (n=70). We found that the healthy sibling group was impaired in tests measuring performance speed and executive functions. The patients were significantly impaired in all neuropsychological variables studied when compared with the control subjects, and also when compared with the healthy siblings. The effects of age, sex and education were controlled for. In conclusion, in a study of representative, population-based sample the healthy siblings of schizophrenia patients demonstrated deficits in processing speed and executive functions.