Biomedical subjects

Allan Green

Publications and source records attributed to Allan Green.

18 recordsLinked to original sources

PubMed2006

99mTc-HYNIC-annexin V SPECT imaging of acute stroke and its response to neuroprotective therapy with anti-Fas ligand antibody.

PURPOSE: The first aim of the study was to determine whether (99m)Tc-HYNIC-annexin V, a marker of cellular stress and apoptosis, can detect ischemic injury in patients with acute stroke. Secondly, we wished to test radiolabeled annexin's ability to monitor therapy in a rodent model of focal ischemic injury. METHODS: SPECT imaging of patients was performed between 1 and 2 h after intravenous injection of 30 mCi (1,110 MBq) of tracer. Eight MFL4 (anti-FasL) antibody-treated (400 microg i.p. days 0 and 3) and 21 control adult male Sprague-Dawley rats underwent small animal SPECT imaging with 5-10 mCi (185-370 MBq) of tracer, 1 and 6 days after a 2-h intraluminal thread occlusion of the left middle cerebral artery. RESULTS: Two patients with acute stroke had regions of multifocal annexin uptake that correlated with sites of restricted diffusion on MRI. Anti-FasL antibody treatment significantly reduced annexin uptake by 92% with a 60% decrease in the number of caspase-8 staining (apoptotic) neurons on day 1. On day 6, treated animals had an 80% reduction in tracer uptake with a 75% decrease in infarct size as compared with controls. Annexin uptake in controls and treated animals (day 6) linearly correlated with infarct size (r (2)=0.603, p=0.0036) and the number of TUNEL-positive (apoptotic) nuclei (r (2)=0.728, p=0.00084). CONCLUSION: Annexin imaging shows foci of increased uptake at sites of ischemic injury in patients with acute stroke. Annexin imaging can assess the effects of therapy for ischemic cerebral injury in rats, suggesting its potential as a non-invasive indicator of drug efficacy in future clinical trials.

Acute Disease↗

PubMed2006

Effects of fenofibrate on lipid metabolism in adipose tissue of rats.

The effect of fenofibrate, a peroxisome proliferator-activated receptor alpha agonist, on body weight gain and on reduction of adipose tissue pads has been ascribed to increased fat catabolism in liver mainly through the induction of target enzymes involved in hepatic lipid metabolism. The aim of this study was to investigate whether peroxisome proliferator-activated receptor alpha activation also affects metabolic pathways in adipose tissue of rats treated with fenofibrate (100 mg/kg body weight) for 9 days. Fenofibrate lowered body weight gain and plasma triglyceride, total cholesterol, and high-density lipoprotein cholesterol but had no influence on food intake and on plasma glucose levels. The activity of lipoprotein lipase of treated animals decreased 50% in epididymal, 29% in retroperitoneal, and was not affected in the mesenteric fat pads. In this study, we show a 34% decrease in epididymal adipose tissue de novo lipogenesis by fenofibrate. The results demonstrate that insulin sensitivity of lipolysis is decreased in fenofibrate-treated rats which resulted in 30% higher rate of glycerol release when compared to the control group. These findings suggest that besides its effects on liver, fenofibrate exerts effects on lipid metabolism in adipose tissue which may contribute to decreasing adiposity.

Adipose Tissue↗

PubMed2006

Transferrin and iron induce insulin resistance of glucose transport in adipocytes.

Normal serum can increase the rate of lipolysis in isolated adipocytes. Recently, we reported that the lipolytic effect of serum could be partly explained by effects of iron and transferrin. To further investigate these effects on fat cell metabolism, we have investigated effects of serum, iron, and transferrin on glucose transport in isolated rat adipocytes. Adipocytes were isolated by collagenase digestion of rat epididymal fat pads, and glucose transport was measured as uptake of [3H]2-deoxyglucose, measured in the presence of 0 to 25 ng/mL insulin. Insulin stimulated glucose transport approximately 8- to 10-fold, with a half-maximally effective concentration (EC50) of approximately 0.15 ng/mL. This was not affected by 45-minute treatment with normal human serum. However, when adipocytes were incubated with serum for 4 hours, cells became markedly insulin resistant. This was manifested as decrease in maximally stimulated glucose transport and a rightward shift in the dose-response curve. Both FeS04 (3 microg/mL) and transferrin (100 microg/mL) had similar, although less pronounced effects on insulin-stimulated glucose transport. Treatment of adipocytes with palmitic acid (120 micromol/L), representing the concentration of fatty acids released into the media after 4 hours of serum treatment, did not alter the effect of insulin on glucose transport. We conclude that transferrin and iron induce insulin resistance of glucose transport in adipocytes through a mechanism independent of fatty acids. These findings may further explain the association between body iron stores and risk of type 2 diabetes mellitus.

Adipocytes↗

PubMed2006

Metaplastic breast carcinoma with osseous differentiation: a case report.

Metaplastic breast carcinoma is a rare entity with the distinguishing feature of having epithelial and mesenchymal tissue types incorporated within one tumor. This is a case report of a patient found to have a rare metaplastic breast carcinoma with prominent osseous differentiation. Radiologic and pathologic correlation is provided.

Breast↗

PubMed2005

Chronic disease risk in central New York dairy farmers: results from a large health survey 1989-1999.

BACKGROUND: The agricultural workplace presents a variety of health and safety hazards; it is unknown whether farm work may be a risk factor for certain chronic diseases. METHODS: The health survey data from a large rural population in central New York were used from two studies (1989, 1999) to assess both 1999 prevalence and 10-year incidence of self-reported diabetes, heart disease, hypercholesterolemia, and hypertension among farm (predominantly dairy) and non-farm residents. The 1999 asthma prevalence was also assessed. RESULTS: Multiple logistic regression models for 1999 prevalence found statistically significant protective effects of farming for hypertension (OR=0.83, P=0.0105) and hypercholesterolemia (OR=0.853, P=0.0522). Non-significant results were seen for heart disease (OR=0.67, P=0.128) and diabetes (OR=0.856, P=0.1358). The model for 1999 asthma prevalence showed a significantly elevated risk for farming (OR=1.542, P=0.0004). Logistic models created for the 10-year incidence of hypertension, hypercholesterolemia, diabetes, and heart disease did not show a significant effect for farming. CONCLUSIONS: The protective effect of farming observed for the 1999 prevalence of hypertension and hypercholesterolemia was not seen for the 10-year incidence of these diseases.

Agricultural Workers' Diseases↗

PubMed2004

Assessment of coverage rates and bias using double sampling methodology.

OBJECTIVE: In 1999, a survey of the health status of six rural central New York counties was performed using double sampling. The resulting impact of the methodology on health outcome prevalence estimation was assessed. STUDY DESIGN AND SETTING: Six counties were double sampled using methodology described by Neyman. Prevalence of health outcomes was estimated for the initial survey responders, nonresponders who subsequently responded after being offered a financial incentive, and both groups combined. RESULTS: Although both groups were similar on demographic variables, they differed significantly on several disease and health behavior prevalences. The failure to use double sampling would have resulted in overestimation of the prevalence of chronic disease by as much as 6.2% for females and 2.0% for males. CONCLUSION: Double sampling increased the generalizability of the results from approximately 35% of the population to over 70%, and decreased the bias in the estimation of many of the health end points.

Adult↗

PubMed2004

Fibromatosis: the breast cancer imitator.

Fibromatosis of the breast is a rare benign tumor that should be included in the differential diagnosis for breast cancer. It is usually indistinguishable from malignancy on ultrasound, mammography, physical examination, and on gross evaluation. Distinction is easily made by histologic findings. This benign tumor does not metastasize, but is locally aggressive and tends to recur postoperatively, which accounts for considerable morbidity. We present two cases and a discussion from the perspective of the radiologist, the surgeon, and the pathologist.

Adult↗

PubMed2004

The imaging of apoptosis with the radiolabeled annexin V: optimal timing for clinical feasibility.

In recent years, the imaging of drug-induced apoptosis has become one of the centers of interest in experimental and clinical research. In particular, the accurate monitoring of chemosensitivity as well as the early prediction of chemoresistance in response to various pro-apoptotic interventions are critical requirements for the best management of oncology patients. The use of technetium [(99m)Tc]-labeled annexin V on animal and human models of cancers provides a proof of principle for the feasibility of a non-invasive, in vivo detection of an apoptotic signal and then for the early assessment of tumor response in the course of chemotherapy. Although promising, however, the initial clinical data point out on the technical limitations that are still to be resolved in terms of tumor-to-background ratio and optimal timing for the imaging of apoptosis. In the present review article, we report the results of animal studies aimed to the evaluation of apoptotic peaks following chemotherapy. In the light of these basic research works, we analyze the profiles of radiolabeled annexin V uptake over time as observed in clinical trials. We then discuss possible new imaging strategies designed to optimize the visualization of apoptotic changes within tumor tissues using the [(99m)Tc]-labeled annexin V. We also suggest longer lived forms of radiolabeled annexin V designed to better understand the temporal patterns of apoptotic tumor response, which in turn, may help to capture the best time-window for the imaging of cell death.

Animals↗

PubMed2004

Stimulation of lipolysis by tumor necrosis factor-alpha in 3T3-L1 adipocytes is glucose dependent: implications for long-term regulation of lipolysis.

Tumor necrosis factor-alpha (TNF-alpha) and hyperglycemia both impair insulin sensitivity in vivo. This may be secondary to stimulation of adipose tissue lipolysis and consequent increased circulating free fatty acids (FFAs). Here we report that neither TNF-alpha nor glucose alone has a pronounced effect on lipolysis in 3T3-L1 adipocytes. However, the combination of TNF-alpha plus glucose markedly stimulates lipolysis. Glucose does not affect the ability of isoproterenol to stimulate lipolysis. Alternative substrates such as acetate, pyruvate, and lactate do not allow the TNF-alpha effect. Mannose was almost as effective as glucose; fructose was marginally effective, but galactose was ineffective. The effectiveness of the sugars corresponded with production of lactate, i.e., the cells readily produced lactate from glucose or mannose, slightly from fructose, and not at all from galactose. The ability of TNF-alpha to phosphorylate extracellular signal-regulated kinase 1 (ERK1) and ERK2 and to downregulate perilipin (which has been implicated in the lipolytic effect of TNF-alpha) was not affected by glucose. We conclude that the lipolytic action of TNF-alpha is influenced by glucose in 3T3-L1 adipocytes. The findings suggest that glucose metabolism is required for the lipolytic response to TNF-alpha but not for early signaling events. These findings suggest novel mechanisms by which TNF-alpha and hyperglycemia raise FFA levels and induce insulin resistance.

3T3 Cells↗

PubMed2004

Transferrin and iron contribute to the lipolytic effect of serum in isolated adipocytes.

Previous reports have demonstrated that normal serum can increase the rate of adipocyte lipolysis in vitro. However, the nature of the lipolytic activity has remained obscure. We have investigated the lipolytic activity of human serum using isolated rat adipocytes. Human serum resulted in a dose-dependent stimulation of lipolysis (glycerol release) in adipocytes, with a half-maximal effective dose of 0.05% serum and with maximal stimulation with 1% serum. The effect of serum on glycerol release was rapid (within 30 min), and the effect was reversible. Partial purification of this lipolytic activity using gel filtration and ion-exchange chromatography demonstrates that a protein of approximately 80 kDa contributes to the lipolytic activity. Human transferrin mimicked the activity of partially purified serum, resulting in a maximal 50% increase in basal lipolysis. In addition, ferrous sulfate heptahydrate induced a biphasic increase in the rate of lipolysis, with a maximal increase of 50% at approximately 0.6 microg/ml iron. Inhibitors of protein kinase A (H89) and mitogen-activated protein kinase (PD98059) did not block the effect of serum on lipolysis, whereas the free radical scavenger N-acetyl-l-cysteine completely inhibited the effect. These findings suggest that the stimulatory effect of serum on lipolysis is in part mediated by iron, probably through a prooxidant mechanism.

Adipocytes↗

PubMed2003

Prolonged treatment with prostaglandin E1 increases the rate of lipolysis in rat adipocytes.

Prolonged treatment of adipocytes with certain inhibitors of lipolysis, including N(6)-phenylisopropyl adenosine (PIA) and prostaglandin E(1) (PGE(1)) leads to down-regulation of G(i). Prolonged treatment with PIA increases the rate of lipolysis, and we have reported that tumor necrosis factor-alpha (TNF alpha) stimulates lipolysis by down-regulating G(i). To determine the relationship between G(i) concentration and lipolysis, we have investigated the effect of two other acute inhibitors of lipolysis; PGE(1), which down-regulates G(i), and nicotinic acid (NA), which does not down-regulate G(i). Rat adipocytes were incubated with PIA (300 nM), PGE(1) (3 microM) or nicotinic acid (1 mM) for 24 h. The rate of lipolysis (glycerol release) was increased approximately 2 to 3-fold in PIA-treated cells, and in PGE(1)-treated cells. Conversely, the rate of lipolysis was not altered by the prolonged nicotinic acid treatment. These findings support the hypothesis that the rate of lipolysis in adipocytes is determined, at least partly, by the cellular concentration of G(i) proteins.

Adipocytes↗

PubMed2003

Optimization of the preparation of 99mTc-labeled Hynic-derivatized Annexin V for human use.

Hydrazino nicotinate (Hynic) is one of the most attractive bifunctional agents designed for the labeling of proteins with (99m)Tc. Recently, a (99m)Tc-labeled Hynic-Annexin V derivative has been described and successfully evaluated in animal models of apoptosis. Prior to a phase I human study, the preparation of (99m)Tc-Hynic-Annexin V has been optimized. The influence of the Hynic-load of Annexin V, amount of protein, nature and amount of reducing agent, activity and co-ligand on the labeling yield were evaluated using ITLC and size-exclusion FPLC. Optimal labeling yields were obtained when 60-90 microgram Hynic-Annexin V was labeled with up to 1.11 GBq (30 mCi) (99m)TcO(4)-using 10-20 microgram SnCl(2).2H(2)O as reducing agent and 1.5 mg tricine as the co-ligand. Biodistribution in normal mice was comparable to literature data.

Animals↗

PubMed2003

In vivo imaging of chemotherapy-induced apoptosis in human cancers.

RATIONALE: Induction of apoptosis in sensitive tumor cells is the main mechanism of action of chemotherapy agents in human cancers. Also, the assessment of drug-induced apoptosis soon after chemotherapy may be an early predictor of treatment efficacy. PATIENTS AND METHODS: A phase I/II study was prospectively conducted in 15 patients presenting with proven lung cancers (n = 10), breast cancers (n = 2), and lymphomas (n = 3) to assess the value of the (99m)Tc-radiolabeled recombinant human (rh) Annexin V for imaging apoptosis immediately after completion of the first course of chemotherapy. Early Annexin V findings post-chemotherapy (day+1, day+2) were also compared to the tumor status at 6 to 12 weeks post-treatment. RESULTS: All lung and lymphoma patients with an increased tracer uptake post-treatment (n = 8) had either partial or complete tumor response. Five patients with no tracer uptake had progressive disease. However, two breast cancers had a response to treatment, although no significant tracer uptake was observed. Tumor response and survival time were significantly correlated with the (99m)Tc-labeled Annexin V uptake. No serious events related to tracer administration were noted. CONCLUSION: : Preliminary results of this pilot study demonstrate the feasibility of the 99mTc-labeled Annexin V uptake for the in vivo imaging of apoptosis after one course of chemotherapy. If confirmed on larger series, these promising results may open new perspectives in the management of oncology patients.

Annexin A5↗

PubMed2002

Lipolysis in adipocytes isolated from deep and superficial subcutaneous adipose tissue.

OBJECTIVE: Abdominal subcutaneous adipose tissue (SAT) occurs in two depots separated by a fascial plane: deep SAT and superficial SAT. In a recent study it was demonstrated that the amount of deep SAT has a much stronger relationship to insulin resistance than does superficial SAT. Because insulin resistance may be related to fatty acid release from adipose tissue, we hypothesized that the two SAT depots may have a different lipolytic activity. RESEARCH METHODS AND PROCEDURES: To test this hypothesis, we obtained samples of deep and superficial SAT from patients undergoing elective abdominal surgery. The rate of lipolysis was determined in the collagenase-digested adipocytes obtained from the two fat depots by measuring glycerol release in the presence and absence of isoproterenol. In addition, the relative concentration of hormone-sensitive lipase was determined in both SAT depots by Western blot analysis. RESULTS: Our results showed that the rate of isoproterenol-stimulated lipolysis was approximately 20% higher in cells from deep SAT compared with those from superficial SAT, indicating that the deep SAT is more lipolytically active. The concentration of hormone-sensitive lipase did not differ between the two adipose tissue depots. DISCUSSION: These findings suggest that the higher lipolytic activity of deep SAT may account for its stronger association with insulin resistance. The mechanism seems to be independent of differences in hormone-sensitive lipase concentration.

Adipocytes↗

PubMed2002

Differences in the lipolytic function of adipose tissue preparations from Black American and Caucasian women.

The purpose of this study was to determine the potential causes of the lower lipolytic rates in obese Black American women compared to obese Caucasian women. Subcutaneous and omental adipose tissue were obtained from subjects during abdominal surgery, and hormone-sensitive lipase (HSL) mass, mRNA, and activity were determined. HSL mRNA levels did not differ between the Black American and Caucasian women in either subcutaneous or omental adipose tissue. However, HSL mass was approximately 35% lower (P <.05) in both subcutaneous and omental adipose tissue of the Black Americans. Because of these differences, we measured HSL activity in frozen subcutaneous and omental adipose tissue, and also measured basal and isoproterenol-stimulated lipolytic rates in tissue fragments. No racial differences were found in the activity of HSL in either subcutaneous or omental adipose tissue. However, basal lipolytic rates in the Black Americans were 53% and 44% lower (P <.05) in the subcutaneous and omental fat, respectively, compared to the Caucasian women, despite a lack of difference in cell size between the 2 groups. Interestingly, the degree of stimulation by isoproterenol was higher in both the subcutaneous and omental adipose tissue of the Black American than those of the Caucasian women, resulting in equal stimulation by isoproterenol in the 2 groups. These results indicate that despite the lower mass and lower basal HSL activity in the obese Black American women, stimulation of HSL results in equal activity of the enzyme in the 2 races. This suggests that the signaling pathway of HSL stimulation is more efficient in the Black American women.

Adipocytes↗

PubMed2002

High-oleic and high-stearic cottonseed oils: nutritionally improved cooking oils developed using gene silencing.

Gene technology and plant breeding are combining to provide powerful means for modifying the composition of oilseeds to improve their nutritional value and provide the functional properties required for various food oil applications. Major alterations in the proportions of individual fatty acids have been achieved in a range of oilseeds using conventional selection, induced mutation and, more recently, post-transcriptional gene silencing (PTGS). In particular, a number of high-oleic oils have been developed in order to provide high-stability cooking oils. These oils provide the opportunity to replace the current widespread use of saturated fats and hydrogenated oils that contribute significantly to increased risk of cardiovascular disease due to the effect of saturated and trans-fatty acids on elevating LDL cholesterol in the bloodstream. Similarly, oils with increased stearic acid content are being developed to enable the production of solid fats without the need for hydrogenation. We have recently applied hpRNA-mediated PTGS in cotton to down-regulate key fatty acid desaturase genes and develop nutritionally-improved high-oleic (HO) and high-stearic (HS) cottonseed oils (CSOs). Silencing of the ghFAD2-1 delta12-desaturase gene raised oleic acid content from 13% to 78% and silencing of the ghSAD-1 delta9-desaturase gene substantially increased stearic acid from the normal level of 2% to as high as 40%. Additionally, palmitic acid was significantly lowered from 26% to 15% in both HO and HS lines. Intercrossing the HS and HO lines resulted in a wide range of unique intermediate combinations of palmitic, stearic, oleic and linoleic contents. The oxidative stability, flavor characteristics and physical properties of these novel CSOs are currently being evaluated by food technologists.

Cottonseed Oil↗

PubMed2002

Insulin and rosiglitazone regulation of lipolysis and lipogenesis in human adipose tissue in vitro.

Lipolysis is an important process determining fuel metabolism, and insulin regulates this process in adipose tissue. The aim of this study was to investigate the long-term effects of insulin, an insulin enhancer (rosiglitazone [RSG]), and insulin in combination with RSG on the regulation of lipolysis and lipogenesis in human abdominal subcutaneous fat. Lipolysis and lipogenesis were assessed by protein expression studies of hormone-sensitive lipase (HSL) (84 kDa) and lipoprotein lipase (LPL) (56 kDa), respectively. In addition, lipolytic rate was assessed by glycerol release assay and tumor necrosis factor (TNF)-alpha release measured by enzyme-linked immunosorbent assay (n = 12). In subcutaneous adipocytes, increasing insulin doses stimulated LPL expression, with maximal stimulation at 100 nmol/l insulin (control, 1.0 +/- 0.0 [mean +/- SE, protein expression relative to control]; 1 nmol/l insulin, 0.87 +/- 0.13; 100 nmol/l insulin, 1.68 +/- 0.19; P < 0.001). In contrast, insulin at the 100 nmol/l dose reduced the expression of HSL (100 nmol/l insulin, 0.49 +/- 0.05; P < 0.05), while no significant reduction was observed at other doses. Higher doses of insulin stimulated both HSL (1,000 nmol/l insulin, 1.4 +/- 0.07; P < 0.01) and LPL (control 1.00 +/- 0.0; 1,000 nmol/l insulin, 2.66 +/- 0.27; P < 0.01) protein expression. Cotreatment with RSG induced an increased dose response to insulin for LPL and HSL (P < 0.05); RSG alone also increased LPL and HSL expression (P < 0.05). Insulin stimulated TNF-alpha secretion in a dose-dependent manner (P < 0.01); the addition of RSG (10(-8) mol/l) reduced TNF-alpha secretion (P < 0.05). In summary, chronic treatment of human adipocytes with insulin stimulates lipolysis and LPL protein expression. The addition of RSG reduced the lipolytic rate and TNF-alpha secretion. The increase in lipolysis is not explained by changes in HSL expression. These data, therefore, may explain in part why hyperinsulinemia coexists with increased circulating nonesterified free fatty acids and increased adiposity in obese and/or type 2 diabetic patients.

Adipocytes↗

PubMed2002

Increased uptake of the apoptosis-imaging agent (99m)Tc recombinant human Annexin V in human tumors after one course of chemotherapy as a predictor of tumor response and patient prognosis.

PURPOSE: Many anticancer therapies exert their therapeutic effect by inducing apoptosis in target tumors. We evaluated in a Phase I study the safety and the feasibility of (99m)Tc-Annexin V for imaging chemotherapy-induced apoptosis in human cancers immediately after the first course of chemotherapy. EXPERIMENTAL DESIGN: Fifteen patients presenting with lung cancer (n = 10), lymphoma (n = 3), or breast cancer (n = 2) underwent (99m)Tc-Annexin V scintigraphy before and within 3 days after their first course of chemotherapy. Tumor response was evaluated by computed tomography and (18)F-fluoro-2-deoxy-D-glucose positron emission tomography scans, 3 months in average after completing the treatment. Median follow-up was 117 days. RESULTS: In all cases, no tracer uptake was observed before treatment. However, 24-48 h after the first course of chemotherapy, 7 patients who showed (99m)Tc-Annexin V uptake at tumor sites, suggesting apoptosis, had a complete (n = 4) or a partial response (n = 3). Conversely, 6 of the 8 patients who showed no significant posttreatment tumor uptake had a progressive disease. Despite the lack of tracer uptake after treatment, the 2 patients with breast cancer had a partial response. Overall survival and progression-free survival were significantly related to tracer uptake in treated lung cancers and lymphomas (P < 0.05). No serious adverse events were observed. CONCLUSIONS: Our preliminary results demonstrated the feasibility and the safety of (99m)Tc-Annexin V for imaging apoptosis in human tumors after the first course of chemotherapy. Initial data suggest that early (99m)Tc-Annexin V tumor uptake may be a predictor of response to treatment in-patients with late stage lung cancer and lymphoma.

Annexin A5↗