Antitumor activity of the angiogenesis inhibitor TNP-470 on murine lymphoma/leukemia cells in vivo and in vitro

OBJECTIVE: The aim of this study was to evaluate the effects of an angiogenesis inhibitor in a non-immunocompromised setting in which transplanted tumor cells home and expand in a manner mimicking the original tumor in the donor. We used a novel animal model for T-cell lymphoma/leukemia (TLL) to test the antitumor effect of TNP-470, a well-established angiogenesis inhibitor. MATERIALS AND METHODS: Cells from spontaneously arisen TLL tumors were transferred to syngenic recipients. The mice were treated with TNP-470 (30 mg/kg) or vehicle every other day for 2 weeks. Mice were sacrificed on day 15 after transfer, and body and organ weights were measured. Cell cycle and morphologic analysis was performed on cells and/or sections from selected organs. The cytotoxic effect of TNP-470 was assayed in vitro using the TLL-M and HL-60 cell lines. RESULTS: TNP-470 treatment significantly reduced total tumor load and tumor mass in specific organs infiltrated with lymphoma/leukemia. This was associated with an increased apoptosis in these organs. We also observed side effects of TNP-470 not previously reported, such as diminished extramedullary erythropoiesis and disrupted liver morphology. In vitro TLL-M cells were resistant to cytotoxic effects of moderate doses of TNP-470. CONCLUSIONS: TNP-470 treatment has a beneficial effect on tumor load in the TLL transfer model, most likely caused by the antiangiogenic effect of TNP-470. This is supported by the observation of increased apoptosis in infiltrated organs. The TLL transfer model is well suited for further studies of combinations with TNP-470 or other angiogenesis inhibitors and cytotoxic drugs.     

Effectiveness and safety of strengthening,aerobic, and coordination exercises for patients with osteoarthritis

Exercise has generated interest as an important nonpharmacological treatment for symptomatic osteoarthritis (OA) of the hip and knee. Effect sizes in exercise interventions are small to moderate for pain and functional improvements and are similar to those observed for improvement in pain for nonsteroidal anti-inflammatory drugs (NSAIDS). However, in contrast to NSAIDS, exercise interventions are safe and improve function through a direct effect on muscle strength and function. Both aerobic and strengthening exercises seem to be equally effective in regard to pain and function in patients with OA. In obese patients with OA, a combination of diet and exercise may be advantageous for optimal benefits in health-related quality-of-life and physical function. This article will focus on recent (September 2001-August 2002) randomized controlled trials with exercise as an intervention in patients with OA.     

Biochemical and structural analysis of the NS5B RNA-dependent RNA polymerase of the hepatitis C virus

Hepatitis C virus (HCV), the major causative agent of chronic and sporadic non-A, non-B hepatitis worldwide, is a distinct member of the Flaviviridae virus family. These viruses have in common a plus-strand RNA genome that is replicated in the cytoplasm of the infected cell via minus-strand RNA intermediates. Owing to the lack of reliable cell culture systems and convenient animal models for HCV, the mechanisms governing RNA replication are not known. As a first step towards the development of appropriate in vitro systems, we expressed the NS5B RNA-dependent RNA polymerase (RdRp) in insect cells, purified the protein to near homogeneity and studied its biochemical properties. It is a primer- and RNA template-dependent RNA polymerase able to copy long heteropolymeric templates without additional viral or cellular cofactors. We determined the optimal reaction parameters, the kinetic constants and the substrate specificity of the enzyme, which turned out to be similar to those described for the 3D polymerase of poliovirus. By analysing a series of nucleosidic and non-nucleosidic compounds for their effect on RdRp activity, we found that ribavirin triphosphates have no inhibitory effect, providing direct experimental proof that the therapeutic effect observed in patients is not related to a direct inhibition of the viral polymerase. Finally, mutation analysis was performed to map the minimal NS5B sequence required for enzymatic activity and to identify the 'classical' polymerase motifs important for template and NTP binding and catalysis.     

Hypertension in beta-adducin-deficient mice

Polymorphic variants of the cytoskeletal protein adducin have been associated with hypertension in humans and rats. However, the direct role of this protein in modulating arterial blood pressure has never been demonstrated. To assess the effect of beta-adducin on blood pressure, a beta-adducin-deficient mouse strain (-/-) was studied and compared with wild-type controls (+/+). Aortic blood pressure was measured in nonanesthetized, freely moving animals with the use of telemetry implants. It is important to note that these mice have at least 98% of C57Bl/6 genetic background, with the only difference from wild-type animals being the beta-adducin mutation. We found statistically significant higher levels of systolic blood pressure (mm Hg) (mean+/-SE values: -/-: 126.94+/-1.14, n=5; +/+: 108.06+/-2. 34, n=6; P:相似文献   

Antibody response to a synthetic peptide covering a LAV-1/HTLV-IIIB neutralization epitope and disease progression

Sequential sera of homosexual men participating in a prospective study on the incidence of HIV-1 infection and risk factors for AIDS were tested for the presence of antibodies to a synthetic 17-mer (Neu21; KSIRIQRGPGRAFVTIG) representing a neutralization epitope as present on the LAV-1/HTLV-IIIB strain of HIV-1. Of 191 at entry, 143 (75%) HIV-1-seropositive men remained anti-Neu21-negative during the follow-up period of 36 months. Thirty-seven (19%) HIV-1-seropositive men were persistently anti-Neu21-positive. Eleven (6%) HIV-1 seropositive men seroconverted for anti-Neu21 during follow-up. Of 75 men developing antibodies to HIV-1, 17 (23%) developed antibodies to Neu21. The incidence of anti-Neu21 seroconversion (calculated as attack rate) after 36 months was significantly lower (P less than 0.00001) among HIV-1-seropositive individuals (8%) than among the 75 HIV-1 seroconverters tested (25%), indicating that seroconversion to this neutralization domain occurs early during infection. AIDS and AIDS-related complex were diagnosed in 17% of the anti-Neu21 negatives and in 11% of the anti-Neu21 positives; this difference was not significant. The presence of HIV-1 antigen (29 versus 28%), the absence of antibodies to core proteins (45 versus 46%) and low CD4+ cell numbers (34 versus 40%) were not seen more frequently among anti-Neu21 negatives than among anti-Neu21 positives. These results argue against a role for antibodies to this LAV-1/HTLV-IIIB neutralization domain in protection against disease progression.     

Thyroid peroxidase antibodies, levels of thyroid stimulating hormone and development of hypothyroidism in euthyroid subjects

ObjectiveThyroid peroxidase antibodies (TPOAbs) have been found to be related to the levels of thyroid stimulating hormone (TSH) and to predict future development of thyroid failure in selected populations. We investigated these relations in a euthyroid general population.DesignCross-sectional investigation of the relationship of TPOAbs and levels of TSH in euthyroid subjects. Prospective investigation of the association of TPOAbs and TSH with development of hypothyroidism. Incident hypothyroidism was defined as initiation of l-thyroxine in the absence of thyreostatic medication.SubjectsThe study was performed in a random sample of 2703 participants of the PREVEND study. A total of 309 subjects were excluded from analyses, mainly because of TSH outside the reference range (0.35–4.94 mIU/l; n = 115).ResultsMean (SD) baseline age was 47.7 (12.5) years, with 50.8% females. Prevalence of positive TPOAbs (≥ 12 kU/l) was 8.4%. TSH concentrations were increased in subjects with TPOAbs (P < 0.001). During a median follow-up of 9.1 years, 15 (0.6%) subjects developed hypothyroidism (3.5% in TPOAbs positive vs. 0.4% in TPOAbs negative subjects; P < 0.001). Female sex (P = 0.02), and TSH (P < 0.001) were also significantly associated with incident hypothyroidism. In multivariate analysis, TSH and TPOAbs remained independent predictors (both P < 0.001).ConclusionsWe confirmed the positive relationship of the presence of TPOAbs with levels of TSH and showed that TPOAbs and TSH predict future development of hypothyroidism. These results are consistent with the presence of TPOAbs necessitating a compensatory increase in levels of TSH for maintenance of euthyroidism, even in the euthyroid range.     

Psychosocial and geriatric correlates of functional status after total hip replacement

OBJECTIVE: To determine whether psychosocial factors, chronic diseases, and common geriatric problems are associated with poor physical function 3 years after primary total hip replacement (THR). METHODS: We studied a sample of Medicare recipients in Ohio, Pennsylvania, and Colorado (n = 922) who underwent primary THR in 1995 (mean +/- SD age 73.1 +/- 5.6 years, 32% men). Participants completed a questionnaire regarding lifestyle factors, medical history, and quality of life approximately 3 years after the surgery. Physical function was measured using the function subscale of the Western Ontario and McMaster Universities Osteoarthritis Index. We assessed the relationship between functional outcome 3 years postsurgery and 4 predictor domains: pain or complications in the operated hip, other musculoskeletal comorbidity, medical factors (obesity, chronic medical comorbidity, rheumatoid arthritis, and such common geriatric problems as falls, poor balance, or incontinence), and psychosocial factors (mental health, regular alcohol consumption, smoking, provider role, living alone, and education). RESULTS: Ten percent of subjects had poor functional status. In a logistic regression model controlling for sex and age, the following factors were associated with an increased risk for poor functional status (in order of importance): pain in the back or lower extremity, severe pain in the operated hip, poor mental health, more than 1 common geriatric problem, obesity, and less than college education. CONCLUSION: Pain in the operated hip was strongly associated with poor functional status 3 years after THR. However, other factors associated with poor functional status were not related to the hip. Our results suggest that a comprehensive assessment of functional status in elderly THR patients should include assessment of common geriatric problems, mental health status, and weight.     

A point mutation in gp91-phox of cytochrome b558 of the human NADPH oxidase leading to defective translocation of the cytosolic proteins p47-phox and p67-phox.

The superoxide-forming NADPH oxidase of human phagocytes is composed of membrane-bound and cytosolic proteins which, upon cell activation, assemble on the plasma membrane to form the active enzyme. Patients suffering from chronic granulomatous disease (CGD) are defective in one of the following components: p47-phox and p67-phox, residing in the cytosol of resting phagocytes, and gp91-phox and p22-phox, constituting the membrane-bound cytochrome b558. In an X-linked CGD patient we identified a novel missense mutation predicting an Asp-->Gly substitution at residue 500 of gp91-phox, associated with normal amounts of nonfunctional cytochrome b558 in the patient's neutrophils. In PMA-stimulated neutrophils and in a cell-free translocation assay with neutrophil membranes and cytosol, the association of the cytosolic proteins p47-phox and p67-phox with the membrane fraction of the patient was strongly disturbed. Furthermore, a synthetic peptide mimicking domain 491-504 of gp91-phox inhibited NADPH oxidase activity in the cell-free assay (IC50 about 10 microM), and the translocation of p47-phox and p67-phox in the cell-free translocation assay. We conclude that residue 500 of gp91-phox resides in a region critical for stable binding of p47-phox and p67-phox.