Anti-tumour activity of oncolytic Western Reserve vaccinia viruses in canine tumour cell lines, xenografts, and fresh tumour biopsies.

Cancer is one of the most common reasons for death in dogs. One promising approach is oncolytic virotherapy. We assessed the oncolytic effect of genetically modified vaccinia viruses in canine cancer cells, in freshly excised tumour biopsies, and in mice harbouring canine tumour xenografts. Tumour transduction efficacy was assessed using virus expressing luciferase or fluorescent marker genes and oncolysis was quantified by a colorimetric cell viability assay. Oncolytic efficacy in vivo was evaluated in a nude mouse xenograft model. Vaccinia virus was shown to infect most tested canine cancer cell lines and primary surgical tumour tissues. Virus infection significantly reduced tumour growth in the xenograft model. Oncolytic vaccinia virus has antitumour effects against canine cancer cells and experimental tumours and is able to replicate in freshly excised patient tumour tissue. Our results suggest that oncolytic vaccinia virus may offer an effective treatment option for otherwise incurable canine tumours.

CD40 ligand and tdTomato-armed vaccinia virus for induction of antitumor immune response and tumor imaging.

Oncolytic vaccinia virus is an attractive platform for immunotherapy. Oncolysis releases tumor antigens and provides co-stimulatory danger signals. However, arming the virus can improve efficacy further. CD40 ligand (CD40L, CD154) can induce apoptosis of tumor cells and it also triggers several immune mechanisms. One of these is a T-helper type 1 (Th1) response that leads to activation of cytotoxic T-cells and reduction of immune suppression. Therefore, we constructed an oncolytic vaccinia virus expressing hCD40L (vvdd-hCD40L-tdTomato), which in addition features a cDNA expressing the tdTomato fluorochrome for detection of virus, potentially important for biosafety evaluation. We show effective expression of functional CD40L both in vitro and in vivo. In a xenograft model of bladder carcinoma sensitive to CD40L treatment, we show that growth of tumors was significantly inhibited by the oncolysis and apoptosis following both intravenous and intratumoral administration. In a CD40-negative model, CD40L expression did not add potency to vaccinia oncolysis. Tumors treated with vvdd-mCD40L-tdtomato showed enhanced efficacy in a syngenic mouse model and induced recruitment of antigen-presenting cells and lymphocytes at the tumor site. In summary, oncolytic vaccinia virus coding for CD40L mediates multiple antitumor effects including oncolysis, apoptosis and induction of Th1 type T-cell responses.

Mammal-derived respiratory lipocalin allergens do not exhibit dendritic cell-activating capacity.

Most mammal-derived respiratory allergens belong to the lipocalin family of proteins. Determinants of their allergenic capacity are still unknown. Innate immune cells, in particular dendritic cells, have been shown to be involved in the allergenicity of some proteins. As recognition by dendritic cells is one of the few plausible mechanisms for the allergenicity of proteins, we wanted to investigate their role in the allergenicity of lipocalin allergens. Therefore, we first incubated human monocyte-derived dendritic cells with immunologically functional recombinant allergens mouse Mus m 1, dog Can f 1 and 2, cow Bos d 2, horse Equ c 1 and natural Bos d 2. Then, the surface marker expression and cytokine production of dendritic cells and their capacity to promote T cell proliferation and Th2 immune deviation in naïve CD4(+) T cells were examined in vitro. We found that near to endotoxin-free lipocalin allergens had no effect on the activation, allostimulatory capacity or cytokine production of dendritic cells. The dendritic cells could not induce immune deviation in naïve CD4(+) T cells. In contrast, lipopolysaccharide activated the dendritic cells efficiently. However, lipocalin allergens were not able to modify the lipopolysaccharide-induced responses. We conclude that an important group of mammal-derived respiratory allergens, lipocalins, appear not to be able to activate dendritic cells, a major component involved in the allergenicity of some proteins. It is conceivable that this incapacity of lipocalin allergens to arouse innate immunity may be associated with their poor capacity to induce a strong T cell response, verified in several studies.

Atomistic modeling of metal surfaces under electric fields: direct coupling of electric fields to a molecular dynamics algorithm.

The effect of electric fields on metal surfaces is fairly well studied, resulting in numerous analytical models developed to understand the mechanisms of ionization of surface atoms observed at very high electric fields, as well as the general behavior of a metal surface in this condition. However, the derivation of analytical models does not include explicitly the structural properties of metals, missing the link between the instantaneous effects owing to the applied field and the consequent response observed in the metal surface as a result of an extended application of an electric field. In the present work, we have developed a concurrent electrodynamic-molecular dynamic model for the dynamical simulation of an electric-field effect and subsequent modification of a metal surface in the framework of an atomistic molecular dynamics (MD) approach. The partial charge induced on the surface atoms by the electric field is assessed by applying the classical Gauss law. The electric forces acting on the partially charged surface atoms (Lorentz and Coulomb) are then introduced in the MD algorithm to correct the atomic motion in response to the applied field. The enhancement factor at sharp features on the surface for the electric field and the assessment of atomic charges are discussed. The results obtained by the present model compare well with the experimental and density-functional theory results.

In vivo detection of vascular adhesion protein-1 in experimental inflammation.

Vascular adhesion protein-1 (VAP-1) is an inflammation-inducible endothelial glycoprotein which mediates leukocyte-endothelial cell interactions. To study the pathogenetic significance of VAP-1 in inflammatory disorders, an in vivo immunodetection method was used to detect the regulation of luminally expressed VAP-1 in experimental skin and joint inflammation in the pig and dog. Moreover, VAP-1 was studied as a potential target to localize inflammation by radioimmunoscintigraphy. Up-regulation of VAP-1 in experimental dermatitis and arthritis could be visualized by specifically targeted immunoscintigraphy. Moreover, the translocation of VAP-1 to the functional position on the endothelial surface was only seen in inflamed tissues. These results suggest that VAP-1 is both an optimal candidate for anti-adhesive therapy and a potential target molecule for imaging inflammation.

Active treatment of chronic neck pain: a prospective randomized intervention.

STUDY DESIGN: A randomized comparative study with single-blind outcome assessments. OBJECTIVES: To compare the efficacy of a multimodal treatment emphasizing proprioceptive training (ACTIVE) with activated home exercises (HOME) and recommendation of exercise (CONTROL) in patients with nonspecific chronic neck pain. SUMMARY OF BACKGROUND DATA: The efficacy of active exercises and passive physiotherapy for neck trouble has been somewhat disappointing in the previous few studies. METHODS: Seventy-six patients (22 men, 54 women) with chronic, nonspecific neck pain participated. Sixty-two participated the 1-year follow-up. Subjective pain and disability, cervical ranges of motion, and pressure pain threshold in the shoulder region were measured at baseline, at 3 months, and at 12 months. The ACTIVE treatment consisted of 24 sessions of proprioceptive exercises, relaxation, and behavioral support. The HOME regimen included a neck lecture and two sessions of practical training for home exercises and instructions for maintaining a diary of progress. The CONTROL treatment included a lecture regarding care of the neck with a recommendation to exercise. RESULTS: The average self-experienced total benefit was highest in the ACTIVE group, and the HOME group rated over the CONTROL group (P < 0.001). Differences between the groups in favor of the ACTIVE treatment were recorded in reduction of neck symptoms and improvements in general health and self-experienced working ability (P < 0.01-0.03). Changes in measures of mobility and pressure pain threshold were minor. CONCLUSIONS: Regarding self-experienced benefit, the multimodal treatment was more efficacious than activated home exercises that were clearly more efficacious than just advising. No major differences were noted in objective measurements of cervical function between the groups, but the content validity of these assessments in chronic neck trouble can be questioned.

Resistance training in the treatment of non-insulin-dependent diabetes mellitus.

Aerobic endurance exercise has traditionally been advocated in the treatment of non-insulin-dependent diabetes mellitus (NIDDM). However, the effect of aerobic endurance exercise programs on long-term glycaemic control is small to moderate. The aim of the present study was to determine the effect of circuit resistance training on long-term glycaemic control (HbA1c) and to examine the potential association between muscle size and glycaemic control in NIDDM subjects. Eight NIDDM subjects participated in a 3 month individualized progressive resistance training program (moderate intensity, high-volume) twice a week with measurements of HbA1c, lipids, blood pressure, VO2max and thigh muscle cross-sectional area. There was a significant improvement in HbA1c (8.8% - 8.2%; p < 0.05). Muscle endurance increased by 32 +/- 23% (p < 0.05), and the cross-sectional area of m. vastus lateralis increased by 21% (p < 0.001). There was a strong inverse correlation between HbA1c and muscle cross-sectional area (knee extensors) after the exercise period (r = -0.73; p < 0.05). Circuit resistance training seems to be feasible in moderately obese, sedentary elderly NIDDM subjects and the inclusion of circuit resistance training in exercise training programs for NIDDM subjects should be considered.

Exercise thallium-201 scintigraphy in the localization of myocardial ischaemia.

We performed a retrospective study in order to study the ability of thallium-201 exercise scintigraphy to detect and to localize coronary artery perfusion defects (in comparison with a recent coronary angiogram). We studied 81 patients (67 males); their average age was 52.3 years (men 50.5 and women 54.1 years). They performed a pulse-conducted cycle exercise test, and 2 min before end of exercise 75 MBq of thallium-201 was infused intravenously, and tomographic images were reconstructed by using a Siemens-Rota SPECT gamma camera immediately and 4 h after exercise. The thallium-201 uptake defects were attributed to different coronary arteries, and the results were compared with a coronary angiogram made afterwards in 48 patients. The groups of one-, two- and three-vessel disease were 27, 21, and 21 patients, and only 12 patients did not have significant (over 50%) stenoses. The latter had the highest ejection fraction and working capacity. Sensitivity of thallium-201 exercise scintigraphy was 65%, whereas that of exercise ECG was 41% in patients with a low ejection fraction, while in the whole material the sensitivity of thallium-201 scintigraphy was 91% and that of exercise ECG was 54%. A stenosis in the right coronary artery was best localized by the thallium-201 scintigram (86% correctly); a stenosis in the left anterior descending artery was localized correctly in 75% of the cases, but a stenosis in left circumflex artery was localized correctly only in 44%. We conclude that exercise thallium-201 scintigraphy is a useful method not only in detecting but also in localizing coronary artery disease.

Dynamic (18F)-2-fluoro-2-deoxy-D-glucose (FDG) scintigraphy of normal and tumor-bearing rats.

Five normal rats, 14 rats bearing the Rous sarcoma intrarenally, and four rats with DMBA-induced mammary carcinomas were imaged by dynamic (18F)-2-fluoro-2-deoxy-D-glucose ((18F)FDG) scintigraphy and (99mTc)DTPA renography. The brain, heart, liver, kidneys, and tumor were selected as regions of interest (ROI) for time activity curves; exponential functions were fitted to the decay-corrected curves, which yielded biologic half-lives of (18F)FDG in the ROI. It was found that all organs ROI's exhibited average elimination of activity, whereas activity accumulated in the tumor ROI for the duration of the study (5 h). The (99mTc)DTPA renographies showed that the excretory function in kidneys bearing the Rous sarcoma is severely impaired. This study shows that small laboratory animals can be successfully scintigraphied with a conventional gamma camera using (18F)FDG and that the pharmacokinetics of this radiopharmaceutical may be evaluated.