Exogenous gene expression in tumors: noninvasive quantification with functional and anatomic imaging in a mouse model

PURPOSE: To assess whether a combination of functional (planar imaging and single photon emission computed tomography [SPECT]) and anatomic (magnetic resonance [MR] imaging) imaging techniques can be used to noninvasively quantify tumor expression of a somatostatin receptor type 2A (SSTR2A) gene chimera in vivo. MATERIALS AND METHODS: All animal experiments were approved by the institutional animal care and use committee. Expression of the SSTR2A gene chimera was quantified in vitro, in vivo, and ex vivo. The epitope tag of the fusion protein was detected through an antibody, and the receptor portion was detected by using the Food and Drug Administration-approved radiopharmaceutical indium 111 octreotide. Six mice were injected with cells transfected with vector and with two clonal cell lines that each expressed different amounts of the gene chimera. With a dedicated small-animal gamma camera, planar imaging and SPECT were used for quantification of radiopharmaceutical uptake in vivo; 4.7-T MR imaging was used to derive tumor weight. After imaging, excised tumors were evaluated for uptake and weight. For statistical analysis, linear regression analysis, Wilcoxon rank sum test, and Kruskal-Wallis test were employed. RESULTS: Different expression levels of the chimeric gene were confirmed in vitro. Radiopharmaceutical uptake assessed in excised tumors and that derived from in vivo planar (r = 0.94, P < .05, n = 18) or SPECT (r = 0.90, P < .05, n = 18) images correlated. Weight of excised tumors and that derived from MR images (r = 0.98, P < .05, n = 18) correlated. MR images also allowed morphologic assessment. The biodistribution parameter of percentage of injected dose per gram of excised tumors correlated with the same measure derived from a combination of planar (r = 0.90, P < .05, n = 18) or SPECT (r = 0.87, P < .05, n = 18) images and MR images. CONCLUSION: A combination of noninvasive functional and anatomic imaging can be used in vivo to quantify gene transfer in tumors.     

Deep vein thrombosis: a rare complication of thalidomide therapy in recurrent erythema nodosum leprosum

A case of deep-vein thrombosis is reported in a female patient with multibacillary leprosy who received pulses of dexamethasone and cyclophosphamide for recurrent ENL that had not responded to prednisone and thalidomide.     

Role of bovine adenovirus-3 33K protein in viral replication

The L6 region of bovine adenovirus type (BAdV)-3 encodes a nonstructural protein named 33K. To identify and characterize the 33K protein, rabbit polyclonal antiserum was raised against a 33K-GST fusion protein expressed in bacteria. Anti-33K serum immunoprecipitated a protein of 42 kDa in in vitro translated and transcribed mRNA of 33K. However, three proteins of 42, 38, and 33 kDa were detected in BAdV-3 infected cells. To determine the role of this protein in virus replication, a recombinant BAV-33S1 containing insertional inactivation of 33K (a stop codon created at the seventh amino acid of 33K ORF) was constructed. Although BAV-33S1 could be isolated, the mutant showed a severe defect in the production of progeny virus. Inactivation of the 33K gene showed no effect on early and late viral gene expression in cells infected with BAV-33S1. However, formation of mature virions was significantly reduced in cells infected with BAV-33S1. Surprisingly, insertional inactivation of 33K at amino acid 97 (pFBAV-33.KS2) proved lethal for virus production. Although expression of early or late genes was not affected, no capsid formation could be observed in mutant DNA-transfected cells. These results suggest that 33K is required for capsid assembly and efficient DNA capsid interaction.     

Racial differences in survival among men with prostate cancer and comorbidity at time of diagnosis

OBJECTIVES: This study evaluated the effect of comorbidity at diagnosis on racial differences in survival among men with prostate cancer. METHODS: Clinical and demographic data were abstracted from records of 864 patients diagnosed at 4 Chicago area hospitals between 1986 and 1990. Comorbidity was scored on the basis of clinical information in the Charlson index. Cause-specific relative mortality adjusted for age, stage, differentiation, and treatment was compared across Charlson scores with Cox proportional hazards functions. RESULTS: Blacks had significantly greater mortality from prostate cancer and other causes (vs Whites, relative risk [95% confidence interval] = 1.84 [1.22, 2.79] and 1.69 [1.33, 2.29], respectively; P <.001). However, differences disappeared as initial comorbidity increased (1.75 [1.33, 2.31] vs 0.90 [0.59, 1.29] for scores = 0 and > or =5, respectively). CONCLUSIONS: Absence of a significant preexisting medical diagnosis is associated with a higher risk for excess mortality among Black men diagnosed with prostate cancer.     

Reduced and high molecular weight barley beta-glucans decrease plasma total and non-HDL-cholesterol in hypercholesterolemic Syrian golden hamsters

Consumption of concentrated barley beta-glucan lowers plasma cholesterol because of its soluble dietary fiber nature. The role of molecular weight (MW) in lowering serum cholesterol is not well established. Prior studies showed that enzymatic degradation of beta-glucan eliminates the cholesterol-lowering activity; however, these studies did not evaluate the MW of the beta-glucan. The current study was conducted to evaluate whether barley beta-glucan concentrates, partially hydrolyzed to reduce MW, possess cholesterol-lowering and antiatherogenic activities. The reduced MW fraction was compared with a high MW beta-glucan concentrate from the same barley flour. Concentrated beta-glucan preparations were evaluated in Syrian Golden F(1)B hamsters fed a hypercholesterolemic diet (HCD) with cholesterol, hydrogenated coconut oil, and cellulose. After 2 wk, hamsters were fed HCD or diets that contained high or reduced MW beta-glucan at a concentration of 8 g/100 g at the expense of cellulose. Decreases in plasma total cholesterol (TC) and non-HDL-cholesterol (non-HDL-C) concentrations occurred in the hamsters fed reduced MW and high MW beta-glucan diets. Plasma HDL-C concentrations did not differ. HCD-fed hamsters had higher plasma triglyceride concentrations. Liver TC, free cholesterol, and cholesterol ester concentrations did not differ. Aortic cholesterol ester concentrations were lower in the reduced MW beta-glucan-fed hamsters. Consumption of either high or reduced MW beta-glucan increased concentrations of fecal total neutral sterols and coprostanol, a cholesterol derivative. Fecal excretion of cholesterol was greater than in HCD-fed hamsters only in those fed the reduced MW beta-glucan. Study results demonstrate that the cholesterol-lowering activity of barley beta-glucan may occur at both lower and higher MW.     

Optimization of chitosan film as a substitute for animal and human epidermal sheets for in vitro permeation of polar and non polar drugs

The present investigation is aimed at preparing chitosan films capable of simulating the flux of modal drugs, 5-fluorouracil (5-FU) and indomethacin (INDO), across rat, rabbit and human cadaver epidermal sheets. Application of statistical design revealed that the concentration of chitosan, crosslinking time and concentration of crosslinking agent significantly influenced the in vitro flux of 5-FU and INDO across chitosan films. Multiple linear regression revealed a linear influence of all these active variables on 5-FU and INDO flux. It was deduced from atomic absorption spectroscopic analyses, DSC and IR spectroscopic data that 5% (m/V) sodium tripolyphosphate (NaTPP) produced optimum crosslinking of chitosan films. The in vitro permeation of both 5-FU and INDO across optimized film formulations was found to be comparable to that obtained across rat, rabbit and human epidermal sheets. These results indicate that optimized chitosan films have a potential to be developed as a substitute for animal and human cadaver epidermal sheets for preliminary in vitro permeation studies.     

Protective effect of naringin, a bioflavonoid on ferric nitrilotriacetate-induced oxidative renal damage in rat kidney

An iron chelate, ferric nitrilotriacetate (Fe-NTA), induces acute proximal tubular necrosis as a consequence of lipid peroxidation and oxidative tissue damage that eventually leads to high incidence of renal adenocarcinomas in rodents. This study was designed to investigate the effect of Naringin, a bioflavonoid with anti-oxidant potential, on Fe-NTA-induced nephrotoxicity in rats. One hour after a single intra-peritoneal (i.p.) injection of Fe-NTA (8 mg iron/kg body weight), a marked deterioration of renal architecture and renal function was observed. Fe-NTA induced a significant renal oxidative stress, demonstrated by elevated thiobarbituric acid reacting substances (TBARS) and reduction in activities of renal catalase, superoxide dismutase, and glutathione reductase. Pre-treatment of animals with Naringin, 60 min before Fe-NTA administration, markedly attenuated renal dysfunction, morphological alterations, reduced elevated TBARS, and restored the depleted renal anti-oxidant enzymes. These results clearly demonstrate the role of oxidative stress and its relation to renal dysfunction and suggest a protective effect of Naringin on Fe-NTA-induced nephrotoxicity in rats.